JP5714745B2 - Quinuclidine compounds as α7 nicotinic acetylcholine receptor ligands - Google Patents

Description

BACKGROUND OF THE INVENTION This disclosure relates generally to compounds and compositions of Formula I, including salts, and methods of using the compounds. The compounds are ligands, agonists and partial agonists for the nicotinic α7 receptor and may be useful in the treatment of various disorders of the central nervous system, particularly affective and neurodegenerative disorders.

  Schizophrenia is a serious mental disorder affecting about 1% of the population. Its progression results in serious dysfunction of mental and social functions, often causing the development of other medical conditions. Susceptibility is often familial and both genetic and environmental factors are considered important. The direct and indirect costs of schizophrenia are estimated to be tens of billions of dollars annually in the United States alone.

  Patients with schizophrenia have an increased risk of suicide (approximately 10% lifetime risk). Patients have 2.5 times higher mortality from all causes, resulting in a 20% shorter life expectancy. The onset of illness can cause a chain of unhealthy lifestyle factors and behaviors that increase the risk of various conditions and consequently increase the risk of death.

  The onset of schizophrenia is most common in late adolescence or early adulthood, with episodes recurring throughout life. Schizophrenia is characterized by the expression of three distinct symptom domains: positive symptoms, negative symptoms, and cognitive symptoms. Psychotic or positive symptoms include delusions, hallucinations, thought disorders and paranoia. Negative symptoms include negative emotions, withdrawal, and loss of pleasure. Cognitive dysfunction includes deficits in attention, working memory and executive function. Although the pathophysiology of schizophrenia is not well understood, most experts consider it a multifactorial disorder in which biological, genetic and environmental factors play a role. Most current treatments target the dopamine system, suggesting that excessive dopamine neurotransmission underlies at least some aspects of schizophrenia. This theory is further supported by the discovery that drugs that increase dopamine levels cause psychosis similar to the positive symptoms of schizophrenia. In addition, post-mortem analysis of brains from patients with schizophrenia shows an increase in the number of D2 dopamine receptors. Although novel antipsychotics known as atypical antipsychotics and active on several additional neurotransmitter receptors have been introduced in the last decade, these drugs also have an effect on D2 dopamine receptors. Sharing. All currently used drugs have significant limitations. Positive symptoms are generally alleviated in most patients, but these medications provide little relief for common and often the most debilitating negative symptoms and cognitive deficits. In addition, antipsychotics have some undesirable and restrictive side effects.

  Nicotine is one of the few drugs that has a positive effect on cognitive function. Many schizophrenic patients smoke, and the smoking rate in patients is 2-4 times that of the general population, with up to 90% of patients who have entered the facility smoking. This smoking habit is characterized as a form of self-treatment.

The nicotinic acetylcholine receptor (nAChR) is a pentameric ligand-gated ion channel that is widely expressed throughout the central and peripheral nervous systems. These channels are fast-desensitizing calcium channels that increase the intracellular concentration of Ca ⁺⁺ ions when opened. Although there are 12 receptors, the most common nicotinic receptors in the brain are α4β2 and α7. The α4β2 complex has been identified as a “high affinity” nicotine site. The homopentameric α7 receptor selectively binds to the natural product α-bungarotoxin, allowing its relatively easy localization and measurement. The α7 receptor is expressed primarily in the cortex, hippocampus and subcortical marginal regions and usually occurs presynaptically. Localization of α7 nAChR to regions related to learning and memory has led to studies using both knockout mice and pharmacological treatments. It is related to sensory gating, memory and neuroplasticity. α7 agonists have been shown to increase the release of rodent neurotransmitters, including dopamine, serotonin, glutamate and GABA. Compounds that selectively bind to α7 receptors, such as α7 agonists and partial agonists, improve learning and memory function, restore scopolamine-induced memory deficits, and cognition induced by NMDA antagonists in normal and aged animals It has been shown to restore defects, restore pharmacologically induced gating functional defects such as amphetamine-induced gating disruption and have some anxiolytic properties. The α7 agonists of the present invention are expected to be useful in the treatment of schizophrenia and cognitive disorders associated with schizophrenia.

  Alzheimer's disease is a progressive neurodegenerative disorder that causes general loss of cognitive function. Incidence increases with age, and it is estimated that 25-50% of people over the age of 85 have some degree of dementia. The diagnosis of Alzheimer's disease implies that the remaining life expectancy is reduced by half compared to normal adults.

  Clinical signs of Alzheimer's disease are progressive cognitive decline, decreased ability to conduct daily life, and changes in neuropsychiatric symptoms or behavior. During the progression of the disease, muscular degeneration and reduced mobility make it impossible for you to take your own food, and ultimately patients are bedridden. The language is very confusing and then completely lost. The patient can no longer perform simple tasks alone and needs constant monitoring. The cost of institutional care accounts for nearly 70% of the cost of disease. Therefore, there is a great need for treatments that improve cognitive function and delay facility accommodation.

  Several studies have shown that Alzheimer's disease is associated with a decrease in nicotinic receptors in the cortex and hippocampus. Nicotine injections or nicotine skin patches have been reported to significantly improve attention, memory, and learning ability in Alzheimer's patients. During the progression of Alzheimer's disease, a progressive loss of nicotine receptors occurs, but relatively more α7 neurons remain compared to the more abundant α4 receptors. Recently, administration of selective nicotinic α7 agonists has been shown to improve cognitive function in Alzheimer's disease patients when administered for as long as 8 weeks. This clinical data is consistent with preclinical data that showed that α7 agonists and partial agonists improved learning and memory function and restored scopolamine-induced memory deficits in normal and aged animals. Accordingly, the compounds of the present invention may be useful for the treatment and prevention of Alzheimer's disease. The amyloid peptide Aβ42 has been shown to bind to the α7 nicotinic receptor (Wang et al., J. Biol. Chem., 2000, 275: 5626-5632; J. Neurochem. 2000, 75: 1155-1161 ). This binding promotes Aβ42 aggregation, is believed to be important for the toxic effects of Aβ42, and may cause deregulation of signaling through the α7 nicotinic receptor. Deletion of the α7 receptor gene improves cognitive impairment 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 Aβ42 and α7 receptors. Treatment with α7 agonists and partial agonists can be an approach for modifying disease in Alzheimer's disease. The α7 receptor can 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 α7 agonists and partial agonists of the present invention may be useful in reducing inflammation in neurodegenerative diseases and disorders such as Alzheimer's disease.

  The α7 receptor has also been shown to be involved in reducing inflammation via the vagus nerve. Furthermore, α7 receptors are expressed in synovial cells from patients with rheumatoid arthritis (RA) and osteoarthritis (OA), and α7 agonists have been shown to inhibit the pro-inflammatory cascade that occurs in rheumatoid joints. (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.

  Nicotinic receptors containing α7 subunits are present in mucosal mast cells known to be involved in gastrointestinal hypersensitivity (Kageyama-Yahara et al., Biochem and Biophys. Research Commun., 2008, v. 377, pp321-325). The α7 agonist GTS-21 inhibits antigen-induced degranulation of mucosal mast cells, suggesting that α7 agonists are useful in the treatment of irritable bowel conditions such as ulcerative colitis.

  In a recent report (Marrero et al., JPET Fast Forward, September 28, 2009, DOI: 10.1124 / jpet.109.154633), an α7 agonist is expressed in a mouse model of type II diabetes (leptin receptor-deficient db / db mice) It has been shown to reduce weight gain, reduce food intake, and reduce elevated plasma levels of triglycerides, glucose, glycated hemoglobin and TNFa. The α7 agonists and partial agonists of the present invention may be useful for the treatment of diabetes.

The following references provide a general review of the nicotine receptor system and the α7 receptor and ligand: 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, pp131-171; Concotta et al., Current Opinion in Investigational Drugs (2008), v 9, pp47-56; Leiser et al., Pharmacol. and Therapeutics (2009), doi: 10: 1016 / j.pharmthera.2009.03.009).

The ligands for the nicotinic α7 receptor are described in the above references, as well as U.S. Patent Application No. 20090270405, U.S. Patent No. 2007004715, International Publication No. 2008/000469, International Publication No. 2003/092580, International Publication No. 2004 / 000,469. No., European Patent No. 337,547, European Patent No. 452,101 and CJ Swain, et al., J. Med. Chem. , (1992) 35: 1019-1031.

  The present invention provides technical benefits, for example, the compounds are novel, are ligands for the nicotinic α7 receptor, and are useful in the treatment of various central nervous system disorders, particularly affective and neurodegenerative disorders obtain. Furthermore, the compounds provide benefits for pharmaceutical applications related to one or more of mechanism of action, binding, inhibitory effect, target selectivity, solubility, safety profile or bioavailability.

DESCRIPTION OF THE INVENTION The present invention encompasses compounds of 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.

[式中、
Ｒ^１は、イソオキサゾリル、ピラゾリル、オキサゾリル、チアゾリル、イミダゾリル、オキサジアゾリル、チアジアゾリル、トリアゾリル、ピリジニル、ピラジニル、ピリダジニル、ピリミジニル、トリアジニル、キノリニル、イソキノリニル、テトラヒドロイソキノリニル、キノキサリニル、キナゾリニル、ナフチリジニル、インダゾリル、インドリル、２−インドロニル(indolonyl)、ベンゾイソオキサゾリル、ベンゾイソチアゾリル、ベンゾオキサゾリル、ベンゾチアゾリル、ベンゾイミダゾリル、フロピリジニル、チエノピリジニル、チエノピリミジニル、イソチアゾロピリジニル、チアゾロピリジニル、チアゾロピリジノニル、チアゾロピラジニル、チアゾロピリミジニル、トリアゾロピリジニル、トリアゾロピラジニル、ピロロトリアジニル、５,６−ジヒドロベンゾ[ｈ]キナゾリニル、５Ｈ−クロメノ[４,３−ｄ]ピリミジニル、６,７−ジヒドロ−５Ｈ−シクロペンタ[ｄ]ピリミジニル、５,６,７,８−テトラヒドロキナゾリニル、７,８−ジヒドロキナゾリン−５(６Ｈ)−オニルおよびテトラヒドロベンゾチアゾリルからなる群から選択され、これらは、Ｃ_１−４アルキル、Ｃ_３−７シクロアルキル、Ｃ_１−４ハロアルキル、Ｃ_１−４アルコキシ、Ｃ_１−４ハロアルコキシ、Ｃ_３−７シクロアルコキシ、Ｃ_１−４アルキルチオ、フェノキシ、ベンジルオキシ、ハロ、ヒドロキシ、シアノ、ニトロ、Ｃ_１−４アルキルスルホニル、ＮＲ^２Ｒ^３、ピロリジノニル、メチレンジオキシ、フリル、チエニル、ピラゾリル、イミダゾリル、チアゾリル、トリアゾリル、ピラジニル、ピリミジニル、ナフチル、Ｃ_１−４アルキルアミド、ＣＯＮＲ^２Ｒ^３、ピリジル、フェニルおよびベンジルからなる群から独立して選択される０〜３個の置換基で置換されており、ここで、イミダゾリル、ピリジル、フェニルおよびベンジルは、ハロ、Ｃ_１−４アルキル、Ｃ_１−４アルコキシ、Ｃ_１−４ハロアルキル、Ｃ_１−４ハロアルコキシおよびＮＲ^２Ｒ^３からなる群から独立して選択される０〜２個の置換基で置換されており;
Ｒ^２は、水素、Ｃ_１−４アルキル、Ｃ_１−４ヒドロキシアルキルまたはＣ_１−４アミノアルキルであって、
Ｒ^３は、水素、Ｃ_１−４アルキル、Ｃ_１−４ヒドロキシアルキルまたはＣ_１−４アミノアルキルであるか；
あるいは、Ｒ_２およびＲ_３は、それらが結合している窒素原子と一体となって、アゼチジニル、ピロリジニル、ピペリジニル、ピペラジニル、Ｎ−(Ｃ_１−４アルキル)ピペラジニル、モルホリニルまたはホモピペリジニルである。]
の化合物もしくはその立体異性体またはそれらの薬学的に許容される塩である。 One aspect of the present invention is a compound of formula I:

[Where
R ¹ is isoxazolyl, pyrazolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, triazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, quinazolinyl, quinazolinyl, naphtholinyl 2-indolonyl, benzoisoxazolyl, benzisothiazolyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, furopyridinyl, thienopyridinyl, thienopyrimidinyl, isothiazolopyridinyl, thiazolopyridinyl, thiazolopyridinyl Nonyl, thiazolopyrazinyl, thiazolopyrimidinyl, triazolopyridinyl, triazolopyrazinyl, pyrrolotri Dinyl, 5,6-dihydrobenzo [h] quinazolinyl, 5H-chromeno [4,3-d] pyrimidinyl, 6,7-dihydro-5H-cyclopenta [d] pyrimidinyl, 5,6,7,8-tetrahydroquinazolyl Selected from the group consisting of nyl, 7,8-dihydroquinazoline-5 (6H) -onyl and tetrahydrobenzothiazolyl, which are C _1-4 alkyl, C _3-7 cycloalkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _1-4 haloalkoxy, C _3-7 cycloalkoxy, C _1-4 alkylthio, phenoxy, benzyloxy, halo, hydroxy, cyano, nitro, C _1-4 alkylsulfonyl, NR ² R ³ Pyrrolidinonyl, methylenedioxy, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, triazolyl, pyrazinyl Pyrimidinyl, _{naphthyl, C 1-4} alkylamido, CONR ² R ^3, pyridyl, which is substituted with 0-3 substituents independently selected from the group consisting of phenyl and benzyl, wherein, imidazolyl, pyridyl , Phenyl and benzyl are independently selected from the group consisting of halo, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 haloalkyl, C _1-4 haloalkoxy and NR ² R ³ 0-2 Substituted with 1 substituent;
R ² is hydrogen, C _1-4 alkyl, C _1-4 hydroxyalkyl or C _1-4 aminoalkyl,
R ³ is hydrogen, C _1-4 alkyl, C _1-4 hydroxyalkyl or C _1-4 aminoalkyl;
Alternatively, R ₂ and R ₃ together with the nitrogen atom to which they are attached are azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, N- (C _1-4 alkyl) piperazinyl, morpholinyl or homopiperidinyl. ]
Or a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

で示される式Ｉの立体異性体である。 Another aspect of the invention is a compound of formula Ia:

Is a stereoisomer of the formula I.

Another aspect of the invention is that R ¹ is 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, furylthiadia Ryl, (dimethylcarboxamide) (methyl) thiazolyl, (pyrrolidinylCO) thiazolyl, phenyltriazolyl, pyridinyl, bromopyridinyl, chloropyridinyl, (chloro) (fluoro) pyridinyl, (chloro) (methyl) pyridinyl, dichloropyridinyl, fluoro Pyridinyl, cyanopyridinyl, (cyano) (methyl) pyridinyl, (cyano) (dimethyl) pyridinyl, methoxypyridinyl, (methylpyrrolidinyl) pyridinyl, phenylpyridinyl, methoxypyridinylpyridinyl, pyridazinyl, bromo Pyridazinyl, chloropyridazinyl, methylpyridazinyl, methoxypyridazinyl, methylthiopyridazinyl, pyrrolidinylpyridazinyl, pyrrolidinylpyridazinyl, phenylpyridazinyl, Pyridinylpyridazinyl, methoxypyridinylpyridazini , 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, methoxy Pyrimidinyl, dimethoxypyrimidinyl, isopropoxypyrimidinyl, cyclopentoxypyrimidinyl, difluoromethoxypyrimidinyl, trifluoroethoxypyrimidinyl, phenoxypyrimidinyl, methylthio Limidinyl, phenylpyrimidinyl, chlorophenylpyrimidinyl, methylphenylpyrimidinyl, methoxyphenylpyrimidinyl, (phenyl) (triazolyl) pyrimidinyl, pyridinylpyrimidinyl, methoxypyridinylpyrimidinyl, methoxypyrimidinylpyrimidinyl, naphthylpyrimidinyl, pyrazinyl, bromopyrazinyl, (bromo) ( Selected from the group consisting of (methoxy) pyrazinyl, chloropyrazinyl, methylpyrazinyl, dimethylpyrazinyl, butylpyrazinyl, cyanopyrazinyl, methoxypyrazinyl, isopropoxypyrazinyl, trifluoromethylpyrazinyl, phenylpyrazinyl and dimethyltriazinyl Or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is that R ¹ is dimethylpyridinoisoxazolyl, benzoxazolyl, chlorobenzoxazolyl, fluorophenylbenzoxazolyl, ethylphenylbenzoxazolyl, dimethylaminophenylbenzoxa Zolyl, pyridinylbenzoxazolyl, benzothiazolyl, acetamidobenzothiazolyl, bromobenzothiazolyl, chlorobenzothiazolyl, (chloro) (methyl) benzothiazolyl, (chloro) (methoxy) benzothiazolyl, fluorobenzothia Zolyl, difluorobenzothiazolyl, cyanobenzothiazolyl, methylbenzothiazolyl, dimethylbenzothiazolyl, (methyl) (methoxy) benzothiazolyl, ethylbenzothiazolyl, trifluoromethylbenzothiazolyl, hydroxy Benzothiazolyl, me Toxoxybenzothiazolyl, ethoxybenzothiazolyl, isopropoxybenzothiazolyl, trifluoromethoxybenzothiazolyl, difluoromethoxybenzothiazolyl, dimethoxybenzothiazolyl, morpholinylbenzothiazolyl, (pyrrolidinyl CO) benzothiazolyl, methylsulfonylbenzothiazolyl, chlorothiazolopyridinyl, dimethylthiazolopyridinyl, benzyloxythiazolopyridinyl, difluoromethoxythiazolopyridinyl, benzotriazolyl, indronyl, indazolyl, Bromoindazolyl, chloroindazolyl, fluoroindazolyl, (methyl) (methoxy) indazolyl, methoxyindazolyl, trifluoromethylindazolyl, trifluoromethoxyindazolyl, difluoromethoxyindazoly , Benzimidazolyl, fluorobenzimidazolyl, methylbenzimidazolyl, (methyl) (methoxy) benzimidazolyl, methoxybenzimidazolyl, tetrahydrobenzothiazolyl, furopyridinyl, dimethylfuroprimidinyl, thienopyrimidinyl, isopropylthienopyrimidinyl, dimethylthienopyrimidinyl, chlorotriazolopyridinyl Methyltriazolopyridinyl, trifluoromethyltriazolopyridinyl, methoxytriazolopyridinyl, triazolopyrazinyl, bromopyrrolotriazinyl, dimethylaminothiazolopyrimidinyl, thiazolopyrazinyl, bromothiazolo Pyrazinyl, methoxythiazolopyrazinyl, methylthiothiazolopyrazinyl, methoxythiazolopyrimidinyl, (methyl) (methoxy) thiazolopyrim Midinyl, quinolinyl, bromoquinolinyl, fluoroquinolinyl, methylquinolinyl, (methyl) (methoxy) quinolinyl, isoquinolinyl, bromoisoquinolinyl, dichloroisoquinolinyl, methylisoquinolinyl, dimethylisoquinolinyl, quinoxalinyl, chloro Quinoxalinyl, 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-dihydroquinazoline-5 (6H) -onyl, A compound of Ia or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is that R ¹ is phenylthiazolyl, (chloro) (methyl) pyridinyl, (bromo) (phenyl) pyrimidinyl, methoxypyrimidinyl, difluoromethoxypyrimidinyl, difluoroethoxypyrimidinyl, cyclopentoxypyrimidinyl, (Methylphenyl) pyrimidinyl, (methoxyphenyl) pyrimidinyl, bromopyrazinyl, chloropyrazinyl, methylthiopyrazinyl, methoxybenzothiazolyl, ethoxybenzothiazolyl, difluoromethoxybenzothiazolyl, thiazolopyridinonyl, trifluoromethylindazo A compound of formula I or Ia or a pharmaceutically acceptable salt thereof selected from the group consisting of ril, benzimidazolyl, isoquinolinyl and quinazolinyl.

Another aspect of the invention is that R ¹ is bromopyridinyl, dichloropyridinyl, methoxypyridinyl, (pyridinyl) pyridinyl, (phenyl) pyrimidinyl, (methoxypyridinyl) pyrimidinyl, (pyrazolyl) pyrimidinyl, (chloropyrazinyl), A compound of formula I or Ia, or a pharmaceutically acceptable salt thereof, selected from the group consisting of (bromo) (chloro) pyrazinyl and chlorobenzothiazolyl.

Another aspect of the invention is that R ¹ is selected from the group consisting of thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, benzothiazolyl, thiazolopyridinyl, indazolyl, benzimidazolyl, isoquinolinyl and quinazolinyl, which are C _{1- 4} alkyl, C _3-7 cycloalkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _1-4 haloalkoxy, C _3-7 cycloalkoxy, C _1-4 alkylthio, phenoxy, benzyloxy, halo, hydroxy , Cyano, C _1-4 alkylsulfonyl, NR ² R ³ , pyrrolidinonyl, methylenedioxy, furyl, thienyl, triazolyl, pyrimidinyl, naphthyl, C _1-4 alkylamide, CONR ² R ³ , pyridyl, phenyl and benzyl Flock Independently is substituted with 0-3 substituents selected, where pyridyl, phenyl and benzyl, _{halo, C 1-4 alkyl, C 1-4 alkoxy, C 1-4} haloalkyl, C _With a compound of formula I or Ia or a pharmaceutically acceptable salt thereof, substituted with 0 to 2 substituents independently selected from the group consisting of _1-4 haloalkoxy and NR ² R ³ is there.

Another aspect of the invention is that R ¹ is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazinyl, thiazolopyridinyl and isoquinolinyl, which are C _1-4 alkyl, C _3-7 cycloalkyl, C _{1 -4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy, C 3-7 cycloalkoxy, C 1-4} alkylthio, phenoxy, benzyloxy, halo, hydroxy, _{cyano, C 1-4} alkylsulfonyl, NR ² R ^3, pyrrolidinonyl, methylenedioxy, furyl, thienyl, triazolyl, pyrimidinyl, _{naphthyl, C 1-4} alkylamido, CONR ² R ^3, pyridyl, 0-3 substituents independently selected from the group consisting of phenyl and benzyl In which pyridyl, phenyl and benzyl But _{halo, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl,} 0-2 substituents independently selected from the group consisting of _{C 1-4} haloalkoxy, and ^NR 2 ^{R 3} Or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is wherein R ¹ is selected from the group consisting of pyridinyl and isoquinolinyl, which are C _1-4 alkyl, C _3-7 cycloalkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _1-4 haloalkoxy, C _3-7 cycloalkoxy, C _1-4 alkylthio, phenoxy, benzyloxy, halo, hydroxy, cyano, C _1-4 alkylsulfonyl, NR ² R ³ , pyrrolidinonyl, methylenedioxy, furyl , Thienyl, triazolyl, pyrimidinyl, naphthyl, C _1-4 alkylamide, CONR ² R ³ , pyridyl, phenyl and benzyl, substituted with 0 to 3 substituents independently selected from in, pyridyl, phenyl and benzyl, _{halo, C 1-4 alkyl, C 1-4} alkoxy _{Shi, C 1-4 haloalkyl, C 1-4} independently from the group consisting of haloalkoxy and ^NR 2 ^{R 3} is substituted with 0-2 substituents selected, the compounds of Formula I or Ia or their The pharmaceutically acceptable salt of

からなる群から選択される化合物またはそれらの薬学的に許容される塩である。 Other aspects of the invention include:

Or a pharmaceutically acceptable salt thereof.

からなる群から選択される化合物またはそれらの薬学的に許容される塩である。 Other aspects of the invention include:

Or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is that R ₁ is 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, wherein each group is optionally C _1-4 alkyl, C _1-4 alkoxy, halogen, hydroxy, cyano, trifluoromethyl, Difluoromethyl, fluoromethyl, Trifluoromethoxy, difluoromethoxy, C _1-4 alkylsulfonyl, furyl, morpholino, methylenedioxy, pyridyl, C _1-4 alkylphenyl, halophenyl, dimethylaminophenyl, C _1-4 alkylamide, —CONR ₂ R ₃ { Where R ₂ and R ₃ are each independently hydrogen, C _1-4 alkyl, hydroxy C _1-4 alkyl, amino C _1-4 alkyl, or R ₂ and R ₃ are Together with the atoms to which is attached is C _3-6 cycloalkyl. }; Phenyl, substituted phenyl, phenylmethyl, substituted phenylmethyl {wherein substituted phenyl and substituted phenylmethyl are selected from the group consisting of halogen, C _1-4 alkyl, C _1-4 alkoxy, trifluoromethyl and trifluoromethoxy. Substituted with independently selected substituents. } A compound of formula I, or a pharmaceutically acceptable salt thereof, substituted with one or two substituents selected from the group consisting of:

For compounds of formula I or Ia, the ranges of all examples of variable substituents including R ¹ , R ² and R ³ can be used independently of any other examples of variable substituents. Therefore, the present invention includes a combination of different aspects.

  Unless otherwise stated, 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 of 2 to 4 carbons with at least one double bond. “Alkynyl” means a straight or branched alkyl group 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. The term having a hydrocarbon moiety (eg, alkoxy) includes straight and branched isomers for the hydrocarbon moiety. The parenthesized terms and the multiple parenthesized terms are intended to clarify the relationship of binding to those skilled in the art. For example, a term such as ((R) alkyl) means an alkyl substituent further substituted with the substituent R.

  The present invention includes all pharmaceutically acceptable salt forms of the compounds. A pharmaceutically acceptable salt is a salt in which the counter ion contributes little to the physiological activity or toxicity of the compound and has the same function as a pharmacological equivalent. These salts can be synthesized using commercially available reactants according to conventional organic techniques. Some anion salt forms are acetate, acistrate, besylate, bromide, chloride, citrate, fumarate, glucuronate, hydrobromide, hydrochloride, iodide Includes hydrates, iodides, lactates, maleates, mesylate, nitrates, pamoates, phosphates, succinates, sulfates, tartrate, tosylate and xinafoate. Some cationic salt forms are ammonium, aluminum, benzathine, bismuth, calcium, choline, diethylamine, diethanolamine, lithium, magnesium, meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium, tromethamine and zinc forms including.

  Some of the compounds of the present invention exist in stereoisomeric forms. The present invention includes all stereoisomeric forms of the compounds, including enantiomers and diastereomers. Methods for synthesizing and separating stereoisomers are known in the art.

The present invention includes all tautomeric forms of the present compounds. Examples of tautomeric pairs are shown below.

The present invention is meant to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. As a general non-limiting example, isotopes of hydrogen include deuterium and tritium. Carbon isotopes include ¹³ C and ¹⁴ C. Isotopically-labeled compounds of the invention are generally customary known to those of ordinary skill in the art using appropriate isotope-labeled reactants in place of unlabeled reactants used at other times. Or can be made by methods similar to those described herein. Such compounds may have a variety of potential uses, such as standards and reagents in measuring biological activity. In the case of stable isotopes, such compounds may have the potential to conveniently modify biological, pharmacological or pharmacokinetic properties.

Synthetic Methods The compounds can be synthesized by methods known in the art, including the methods described below and variations within the skill in the art. Several reactants and intermediates are known in the art. Other reactants and intermediates can be synthesized by methods known in the art using readily available materials. Variables used to describe the synthesis of the compounds (eg, the numbered “R” substituents) are intended only to illustrate the method of synthesizing the compounds and are intended to be claimed or claimed. It should not be confused with variables used in other chapters of the specification. The following methods are for illustrative purposes and are not intended to limit the scope of the invention.

  Some of the compounds can be made using the reactions and methods described in this section. The reaction is carried out in a solvent suitable for the reactants and materials used and is suitable for the transformation to be performed. It will be appreciated by those skilled in the art of organic synthesis that the functional groups present at various positions of the molecule must be compatible with the proposed reactants and reactions. The limitation of substituents that can coexist with such reaction conditions will be readily apparent to those skilled in the art, in which case another method must be used.

Abbreviations used in the schemes generally follow conventions used in the art. The chemical abbreviations used herein and in the examples are defined as follows: “NaHMDS” = bis (trimethylsilyl) amide sodium; “DMF” = N, N-dimethylformamide; “MeOH” = methanol; “Ar” = aryl; “TFA” = trifluoroacetic acid; “LAH” = lithium aluminum hydride; “BOC”, “DMSO” = dimethyl sulfoxide; “h” = hours; “rt” "= room temperature or retention time (according to context);" min "= minute;" EtOAc "= ethyl acetate;" THF "= tetrahydrofuran;" EDTA "= ethylenediaminetetraacetic acid;" _Et 2 O "= diethyl ether;" DMAP " = 4-dimethylaminopyridine; "DCE" = 1,2-dichloroethane; "ACN" = aceto Tolyl; "DME" = 1,2-dimethoxyethane; "HOBt" = 1-hydroxybenzotriazole hydrate; "DIEA" = _{diisopropylethylamine, "Nf" = CF 3 (} CF 2) 3 SO 2 -; and " TMOF ″ = trimethylorthoformate.

Abbreviations used herein are defined as follows: “1 ×” = 1 time, “2 ×” = 2 times, “3 ×” = 3 times, “° C.” = Degrees (Celsius), “ “eq” = equivalent, “g” = gram, “mg” = milligram, “L” = liter, “ml” = milliliter, “μl” = microliter, “N” = specified, “M” = molar concentration, “ “mmol” = mmol, “min” = minute, “h” = hour, “rt” = room temperature, “RT” = retention time, “atm” = atmospheric pressure, “psi” = pounds per square inch ), “Conc.” = Concentration, “sat” or “sat'd” = saturation, “MW” = molecular weight, “mp” = melting point, “ee” = enantiomeric excess, “MS” or “Mass Spec” = Mass spectrometry, “ESI” = electrospray ionization mass spectrometry, “HR” = high resolution, “HRMS” = high resolution mass spectrometry, “LCMS” = liquid chromatography mass spectrometry, “ PLC "= high performance liquid chromatography," RP HPLC "= reverse phase HPLC," TLC "or" tlc "= thin layer chromatography," NMR "= nuclear magnetic resonance spectroscopy," ¹ H "= proton," [delta] " = Delta, “s” = singlet, “d” = doublet, “t” = triplet, “q” = quadruple, “m” = multiplet, “br” = broad, “Hz” = Hertz and “α”, “β”, “R”, “S”, “E” and “Z” are stereochemical symbols well known to those skilled in the art.

Scheme 1

  Compounds of formula I are prepared as shown in Reaction Scheme 1. The ketones of formula III (3-quinuclidone) are known, are commercially available or can 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 cyanide or potassium cyanide + acid. By reaction with a borane / tetrahydrofuran complex, the compound of formula IV can be reduced to the corresponding amino-methyl compound of formula V (borane complex).

  A compound of formula V can be reacted directly with a heteroaryl isothiocyanate in an inert solvent to give a thiourea of formula VI. Alternatively, the heteroarylamine can be reacted with thiocarbonyl-diimidazole to give an activated species that can be used to convert a compound of formula V to a compound of formula VI without isolation. The heteroarylamine can be prepared by methods known to those skilled in the art.

  For example, di-isopropylcarbodiimide can be used to cyclize the thiourea of formula VI to give the oxazoline of formula VII, which is deprotected by treatment with acid to give the racemic final product of the compound of formula I Get. The compounds of formula I can be separated into pure enantiomers of formula Ia and formula Ib by methods known in the art, for example by chiral chromatography.

Scheme 2

  Alternatively, as shown in Reaction Scheme 2, the free amino group of the quinuclidine of formula V can be blocked, for example with carbobenzyloxy chloride (“CBZ-Cl”) to give the compound of formula VIII.

  Racemic compounds of formula VIII can be resolved into their enantiomers, ie, formula IX and formula X, for example by chiral chromatography. Any of the compounds of formula IX or X, preferably the compound of formula X, can be reacted as shown in Reaction Scheme 2.

  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 reduction, to give the chiral quinuclidineamine of formula XI. Similar to Reaction Scheme 1, an amine salt of formula XI can be reacted with an isothiocyanate to give a thiourea of formula VIa, which can be reacted with a dialkylcarbodiimide or mixed thiourea (from reaction with thiocarbonyldiimidazole). The chiral oxazoline quinuclidine compound of formula Ia and its tautomer, compound of formula IIa, can be obtained.

Scheme 2a

  Alternatively, the borane group of formula V can be removed with hydrochloric acid to give the dihydrochloride salt of formula XII, which can be reacted with isothiocyanate in the presence of a base to obtain the intermediate thiourea of formula XIII. This can be cyclized as in Reaction Schemes 1-2 to give compounds of Formula I. Compounds of formula XII may also be prepared by other methods described in US Pat. No. 5,137,895 (8/11/1992).

Scheme 3

  Further, the (hetero) aromatic amine is reacted with carbon disulfide, sodium hydroxide and methyl iodide to give the intermediate, dimethyl carbonimidodithioate of formula XIV. These are reacted with the dihydrochloride of formula XII in the presence of a base to remove 2 mol of methanethiol and directly obtain the desired product of formula Ia.

Biological method
I) α7 nicotinic acetylcholine receptor binding Rat HEK293 cells stably expressing α7 nicotinic acetylcholine receptor (rat α7 nAChR) were used to prepare membranes for binding. At 4 ° C., cells were homogenized in hypotonic lysis buffer consisting of 10 mM Tris (pH 7.4), 5 mM EDTA and protease inhibitors and centrifuged at 32000 × 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 inhibitor and centrifuged at 32000 × g for 20 minutes. The pellet was assayed with 50 mM KH ₂ PO ₄ (pH 7.4, 25 ° C.), 1 mM EDTA, 0.005% Triton-X 100 and 0.1% (v / v) Sigma protease inhibitor cocktail. Suspended again. Aliquots were frozen in dry ice / ethanol and stored at −80 ° C. until the day of assay.

II) Ca ²⁺ sensitive fluorescence-based assay α-7 (FLIPR) for nicotinic acetylcholine receptor channel function in mammalian cells
Summary : Lead compounds are evaluated for agonist activity to α-7, α3β4, α4αβ2 and α1β1δ1ε subtypes of nicotinic ACh receptor ion channels expressed in mammalian HEK 293 cells. Agonist intensity and efficacy are determined from dynamic fluorescence Ca ²⁺ inflow measurements performed using a 384 well FLIPR (fluorescence image plate reader). The usefulness of fluorescent indicators for measuring changes in intracellular divalent cation concentration, especially Ca ²⁺ concentration, in drug discovery activities has been described in detail (Rudiger, R., et al., Nature Reviews , 2003, 4 : 579-586; Gonzalez JE, et al., Receptors and Channels , 2002, 8: 283-295). In this assay, a channel-expressing HEK cell line seeded in a 384-well assay plate incorporates a membrane-permeable fluorescent Ca ²⁺ indicator dye whose green release signal at 510 nm increases in response to increasing intracellular Ca ²⁺ concentration. Make it. The basal fluorescence of the cells is monitored in real time and then the test compound is added acutely. If the compound is an agonist of any of the non-selective cation channels, the channel opens and allows intracellular Ca ²⁺ ions to migrate to the cytoplasm, which binds to the Ca ²⁺ indicator dye and produces a fluorescent output signal. Which is detected by a cooled CCD imaging camera.

Materials and Methods : Reactant: The acetomethoxy (AM) ester of Ca ²⁺ indicator dye Fluo-4 was obtained from InVitrogen (Carlsbad, CA). Acetylcholine and all buffer components were purchased from Sigma Chemical Company, St. Louis, MO. G418 and basal media were purchased from InVitrogen Life Technologies, Carlsbad, CA. Fetal bovine serum was purchased from (InVitrogen, Carlsbad, CA).

Cell culture : HEK-293 cells were grown in basal medium containing 10% (v / v) fetal bovine serum at 37 ° C. in a 5% CO ₂ incubator. HEK-293 cells stably expressing ion channels were grown in the same medium supplemented with 500 μg / ml G418.

Ca ²⁺ flow assay of Ca ²⁺ channels expressed in HEK-293 cells : HEK-293 expressing the ion channel of interest is placed in 20 μl of 10% (on a 384 well black wall clear bottom poly-D-lysine coated plate). v / v) Seeds at a density of about 20,000 cells / well in basal medium containing fetal bovine serum and incubated at 29 ° C. in a 5% CO ₂ incubator for 2 days. Prior to the assay, the cells were loaded with Fluo-4 AM ester. Cellular uptake was 30 μl / well with the culture medium removed and mixed with Hanks balanced salt solution (# 14175-095) containing 20 mM HEPES, 2.5 mM probenecid, 0.5 mM CaCl ₂ , 1 mM MgCl ₂ and 10 μM atropine. This was done by substituting the dye with the AM ester (5 μM). Dye uptake was performed at room temperature for 90 minutes, at which point the dye uptake solution was removed and replaced with 40 μl / well Hanks buffer. Cells that incorporated the dye were mounted on FLIPR384 (Molecular Devices, Sunnyvale, CA). Fluo-4 dye was excited using a 488 nm argon laser line. The emission was filtered using a 540 ± 30 nm bandpass filter. In order to assess the effect of test compounds using the Ca ²⁺ flow assay, the tested compounds were placed in assay prepared plates. The assay was initiated by adding Hanks buffer containing 20 μl / well of test compound in nicotinic receptor ion channel expressing cells. For all assays, data was collected for 10 seconds at 1 Hz (baseline), at which point compound-containing stimulation buffer was added and further measurements were taken at 0.33 Hz for 3 minutes.

Data analysis : Statistical robustness of the nicotine receptor Ca ²⁺ flux assay is determined from blanks and total wells. Total wells define the maximum channel activation (maximum effective dose of acetylcholine) for each compound test plate, and blank wells containing only the corresponding amount of DMSO define zero channel activation. The raw data file of fluorescence units obtained from the FLIPR plate reader is automatically exported and processed by an in-house data analysis tool. Reduced% activation data for each concentration of test compound is fitted using a MathIQ fitting engine (ID Business Solutions Limited, Surrey, UK). Data is analyzed by fitting the change in maximum fluorescence intensity for the Ca ²⁺ flow rate at the indicated conditions of the test compound. Compound intensity (EC ₅₀ value) is calculated from a 20-point concentration response curve (CRC) of the mean of three assay wells. The strength of the test compound (Ymax value) is expressed relative to the maximum response to acetylcholine in all wells.

III) Fos Quantification Assay Male Wistar rats were treated with drugs (1-10 mg / kg) or vehicle (2 ml / kg, subcutaneous). Two hours after the treatment, the rat is quickly decapitated and the brain region of interest is removed on ice, weighed, snap frozen in liquid nitrogen and stored at -80 ° C. Further processing of brain tissue in nuclear extracts and Fos quantification was defined by a commercially available ELISA-based chemiluminescent detection kit (Catalog Number 89860, EZ-Detection c-Fos Transkit, Pierce Biotechnology Inc., Rockford, IL) Follow the protocol.

IV) MK-801 disruption set shift assay in rats This assay uses a modification of the protocol described by Stefani et al. ( Behavioral Neuroscience , 2003, 117: 728-737). In this assay, test compounds were evaluated for their ability to reverse MK-801-induced motor ability deficits (0.03 mg / kg, ip, single dose).

The activities of the specific compounds described herein that were tested in the above assay (II) are shown in Table 1.
Table 1

^a ＥＣ_５０値(ｎＭ)に基づく活性： +++ ＝＜１００ｎＭ； ++ ＝１００〜１０００ｎＭ； + ＝１０００〜１０００００ｎＭ；^b NT ＝ 試験せず； NA ＝ 活性なし(＞１００００００ｎＭ)。

^a Activity based on EC ₅₀ values (nM): +++ = <100 nM; ++ = 100-1000 nM; + = 1000-100,000 nM; ^b NT = not tested; NA = no activity (> 1000000 nM).

Table 1b

^a ＥＣ_５０値(ｎＭ)に基づく活性： +++ ＝＜１００ｎＭ； ++ ＝１００〜１０００ｎＭ； + = １０００〜１０００００ｎＭ; ^b NT ＝試験せず； NA ＝ 活性なし (＞１００００００ｎＭ)。

^a Activity based on EC ₅₀ values (nM): +++ = <100 nM; ++ = 100-1000 nM; + = 1000-100,000 nM; ^b NT = not tested; NA = no activity (> 1000000 nM).

Pharmaceutical Compositions and Methods of Treatment The compounds of formula I bind to α7 and may be useful for treating affective and neurodegenerative disorders. Accordingly, another aspect of the present invention is a composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Another aspect of the invention is the use of a compound of formula I in the manufacture of a medicament for treating affective disorders or neurodegenerative disorders.
Another aspect of the invention is the use of a compound of formula I in the manufacture of a medicament for treating schizophrenia or Alzheimer's disease.

Another aspect of the invention is a method of treating an affective or neurodegenerative disorder, comprising administering to a patient a therapeutically effective amount of a compound of formula I.
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.

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.
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.

Another aspect of the invention is a method of treating cognitive impairment, comprising administering to a patient a therapeutically effective amount of a compound of formula I.
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.

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.
Another aspect of the present invention is a method of treating ulcerative colitis comprising administering to a patient a therapeutically effective amount of a compound of formula I.

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.
Another aspect of the invention is a method of treating diabetes, comprising administering to a patient a therapeutically effective amount of a compound of formula I.

“Patient” means a human who is deemed suitable for treatment by workers in the field of affective and neurodegenerative disorders.
“Treatment”, “treatment” and related terms are used as understood by practitioners in the field of affective and neurodegenerative disorders.

  The compounds of the invention are generally given as a pharmaceutical composition comprising a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, and includes conventional excipients. But you can. Pharmaceutically acceptable carriers are conventional known carriers that have an acceptable safety profile. The compositions encompass all common solid and liquid forms and include, for example, capsules, tablets, lozenges and powders, as well as suspensions, syrups, elixirs and solutions. Compositions are made using common formulation methods, and conventional excipients (eg, binders and wetting agents) and vehicles (eg, water and alcohols) are commonly used in the composition. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, 17th edition, 1985.

  Solid compositions are usually formulated in unit dosage form, with compositions providing about 1-1000 mg of active ingredient per dose being preferred. Some examples of dosages are 1 mg, 10 mg, 100 mg, 250 mg, 500 mg and 1000 mg. In general, other medicaments are present in the same unit dose range as the group of medicaments used clinically. Typically this is between 0.25 and 1000 mg / unit.

  Liquid compositions are usually in unit dosage ranges. In general, liquid compositions are in unit dosage ranges 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. In general, other medicaments are present in a unit dose range similar to the group of medicaments used clinically. Typically this is between 1 and 100 mg / ml.

  The present invention encompasses all conventional administration methods, with oral and parenteral methods being preferred. In general, the dosing regimen is similar to other drugs used clinically. Typically, the daily dose is 1-100 mg / kg body weight per day. In general, many compounds are required orally and less in the parenteral. The specific dosing regime, however, is determined by a physician using sound medical judgment.

Description of specific aspects
¹ H-NMR spectra were measured on a Bruker 500, 400 or 300 MHz instrument and chemical shifts were given in ppm (δ) with reference to tetramethylsilane (δ = 0.0). All evaporations were performed under reduced pressure. Unless otherwise stated, a Shimadzu apparatus using a Phenomenex-Luna 4.6 × 50 mm S 10 reverse phase column at a flow rate of 4 ml / min, 0.1% TFA concentration gradient in methanol / water [0-100%, 3 min, runtime 4 min] with UV detector set at 220 nm or Gemini C18 4.6 × 50 mm 5μ reverse phase column at a flow rate of 5 ml / min with a concentration gradient of 10 mM ammonium acetate acetonitrile / water [ LC / MS analysis was performed (anion mass spectrometry) using a 5-95%, 3 min, runtime 4 min] setting the UV detector at 220 nm. Unless otherwise stated, purification was performed on a Shimadzu high performance liquid preparative chromatography system using a preparative C-18 column with a methanol-water concentration gradient containing 0.1% trifluoroacetic acid (TFA). Using a XTERRA 30 × 100 mm S5 column, the flow rate was 40 ml / min and the concentration gradient was 12 minutes.

Example 1
N- (Benzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３００ml)中のベンゾ[ｄ]チアゾール−２−アミン(２０ｇ, １３３mmol)に、１,１'−チオカルボニルジイミダゾール(３０.８ｇ, １７３mmol)を加えた。反応物を５０℃で２４時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で２時間乾燥させた。生成物であるＮ−(ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(２８.９ｇ, １１１mmol)を、さらに精製することなく次の工程に直接用いた。 Step A: N- (Benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

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 ° C. for 24 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 2 hours. The product N- (benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (28.9 g, 111 mmol) was used directly in the next step without further purification.

Ｎ,Ｎ−ジメチルホルムアミド(１００ml)中のＮ−(ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(９.２ｇ, ３５mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(６.０ｇ, ３５mmol)[Swain C.J., et. al., J. Med. Chem., 35:1019-1031 (1992)に従って合成]を加えた。反応物を６５℃で１５時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質を、フラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル／ヘキサン)、ＴＬＣで検出される最初のスポット／フラクションを生成物として得た。フラクションを合わせて、濃縮し、(３−((３−ベンゾ[ｄ]チアゾール−２−イルチオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを灰白色の粉末として得た(１０.６ｇ, ２９.１mmol, 収率８３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.88 (s, 1 H), 10.30 (s, 1 H), 7.94 (d, J=7.63 Hz, 1 H), 7.55 - 7.74 (m, 1 H), 7.37 - 7.53 (m, J=7.32, 7.32 Hz, 1 H), 7.16 - 7.37 (m, J=7.63, 7.63 Hz, 1 H), 5.39 (s, 1 H), 3.85 (d, 2 H), 2.65 - 3.08 (m, 6 H), 1.99 - 2.22 (m, 1 H), 1.79 - 1.97 (m, 2 H), 1.66 - 1.79 (m, 1 H), 1.08 - 1.63 (m, 4 H). MS (LC/MS) R.T. = 3.40; [M+H]⁺ = 363.1。 Step B: (3-((3-Benzo [d] thiazol-2-ylthioureido) methyl) -3-hydroxy-1-ammonobicyclo [2.2.2] octane-1-yl) trihydroborate

N- (Benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (9.2 g, 35 mmol) in N, N-dimethylformamide (100 ml) was added to (3- (aminomethyl)- 3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate (6.0 g, 35 mmol) [Swain CJ, et. Al., J. Med. Chem ., 35: 1019-1031 (1992)] was added. The reaction was stirred at 65 ° C. for 15 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (50-100% ethyl acetate / hexanes) to give the first spot / fraction as product detected by TLC. Fractions were combined and concentrated to give (3-((3-benzo [d] thiazol-2-ylthioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1- Yl) trihydroborate was obtained as an off-white powder (10.6 g, 29.1 mmol, 83% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.88 (s, 1 H), 10.30 (s, 1 H), 7.94 (d, J = 7.63 Hz, 1 H), 7.55-7.74 (m, 1 H), 7.37-7.53 (m, J = 7.32, 7.32 Hz, 1 H), 7.16-7.37 (m, J = 7.63, 7.63 Hz, 1 H), 5.39 (s, 1 H), 3.85 (d, 2 H), 2.65-3.08 (m, 6 H), 1.99-2.22 (m, 1 H), 1.79-1.97 (m, 2 H), 1.66-1.79 (m, 1 H), 1.08-1.63 (m, 4 H). MS (LC / MS) RT = 3.40; [M + H] ⁺ = 363.1.

Ｎ,Ｎ−ジメチルホルムアミド(１００ml)中の(３−((３−ベンゾ[ｄ]チアゾール−２−イルチオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１０.６ｇ, ２９.１mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(１１.４ml, ７２.８mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、粗製の残渣を得た。少量の酢酸エチル(２０ml)を加え、懸濁液を超音波処理した。固体を濾過し、少量の酢酸エチル(２×１０ml)で洗浄した。固体を真空オーブン(８０℃)中で乾燥させ、(２−(ベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(６.８３ｇ, ２０.８mmol, 収率７２％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.09 (br. s., 1 H) 7.81 (d, J=7.63 Hz, 1 H) 7.63 (d, J=7.93 Hz, 1 H) 7.30 - 7.40 (m, 1 H) 7.15 - 7.24 (m, 1 H) 3.88 (d, J=10.38 Hz, 1 H) 3.77 (d, J=10.38 Hz, 1 H) 3.25 - 3.37 (m, 1 H) 3.17 (dd, J=14.95, 1.83 Hz, 1 H) 2.99 - 3.10 (m, 1 H) 2.79 - 2.98 (m, 3 H) 2.27 (br. s., 1 H) 1.98 - 2.11 (m, 1 H) 1.71 - 1.88 (m, 3 H) 1.45 (br. s., 3 H). MS (LC/MS) R.T. = 2.44; [M+H-BH₃]⁺ = 315.1。 Step C: (2- (Benzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) Trihydroborate

(3-((3-Benzo [d] thiazol-2-ylthioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane in N, N-dimethylformamide (100 ml) To the -1-yl) trihydroborate (10.6 g, 29.1 mmol) was added N, N′-diisopropylcarbodiimide (11.4 ml, 72.8 mmol). The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated to give a crude residue. A small amount of ethyl acetate (20 ml) was added and the suspension was sonicated. The solid was filtered and washed with a small amount of ethyl acetate (2 × 10 ml). The solid was dried in a vacuum oven (80 ° C.) and (2- (benzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2. 2] Octane] -1′-yl) trihydroborate was obtained as a white powder (6.83 g, 20.8 mmol, 72% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.09 (br. S., 1 H) 7.81 (d, J = 7.63 Hz, 1 H) 7.63 (d, J = 7.93 Hz, 1 H) 7.30- 7.40 (m, 1 H) 7.15-7.24 (m, 1 H) 3.88 (d, J = 10.38 Hz, 1 H) 3.77 (d, J = 10.38 Hz, 1 H) 3.25-3.37 (m, 1 H) 3.17 (dd, J = 14.95, 1.83 Hz, 1 H) 2.99-3.10 (m, 1 H) 2.79-2.98 (m, 3 H) 2.27 (br. s., 1 H) 1.98-2.11 (m, 1 H) 1.71-1.88 (m, 3 H) 1.45 (br. S., 3 H). MS (LC / MS) RT = 2.44; [M + H-BH ₃ ] ⁺ = 315.1.

アセトン(９ml)中の(２−(ベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(６.６ｇ, ２０.１mmol)に、３Ｍ ＨＣｌ(５０.３ml, １５１mmol)を加えた。反応物を室温で４時間撹拌した。ＴＬＣによって反応の完了が示された(より低いスポット)。酢酸エチルを加え、水層を分離した。水層を１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×１５０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、白色の粉末を得た。少量の酢酸エチル(２０ml)を粉末に加えた。固体を超音波処理し、濾過し、ラセミのＮ−(ベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(５.１３ｇ, １６.３mmol, 収率８１％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.02 (br. s., 1 H) 7.79 (d, J=7.02 Hz, 1 H) 7.62 (d, J=7.63 Hz, 1 H) 7.29 - 7.38 (m, 1 H) 7.15 - 7.22 (m, 1 H) 3.90 (d, J=10.07 Hz, 1 H) 3.65 (d, J=10.07 Hz, 1 H) 2.98 - 3.10 (m, 2 H) 2.73 - 2.88 (m, 2 H) 2.67 (t, J=7.78 Hz, 2 H) 2.07 (br. s., 1 H) 1.93 (br. s., 1 H) 1.42 - 1.67 (m, 3 H). MS (LC/MS) R.T. = 1.15; [M+H]⁺ = 315.3。 Step D: N- (Benzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (Benzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1 ′ in acetone (9 ml) To -yl) trihydroborate (6.6 g, 20.1 mmol) was added 3M HCl (50.3 ml, 151 mmol). The reaction was stirred at room temperature for 4 hours. TLC showed completion of reaction (lower spot). Ethyl acetate was added and the aqueous layer was separated. The aqueous layer was neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 x 150 ml). The organics were combined, dried over magnesium sulfate, filtered and concentrated in vacuo to give a white powder. A small amount of ethyl acetate (20 ml) was added to the powder. The solid was sonicated, filtered, and racemic N- (benzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] 2-Amine was obtained as a white powder (5.13 g, 16.3 mmol, 81% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.02 (br. S., 1 H) 7.79 (d, J = 7.02 Hz, 1 H) 7.62 (d, J = 7.63 Hz, 1 H) 7.29- 7.38 (m, 1 H) 7.15-7.22 (m, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 2.98-3.10 (m, 2 H) 2.73 -2.88 (m, 2 H) 2.67 (t, J = 7.78 Hz, 2 H) 2.07 (br. S., 1 H) 1.93 (br. S., 1 H) 1.42-1.67 (m, 3 H). MS (LC / MS) RT = 1.15; [M + H] ⁺ = 315.3.

Using a Chiralpak AD-H (3 × 25 cm, 5 μm) column, a mobile phase consisting of CO ₂ /(methanol/ACN/DEA=70/30/0.1 (v / v / v)) = 77/23 The enantiomers were separated. The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (benzo [d] thiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane]- 2-Amine (1.45 g, 4.61 mmol, 29.4% yield).
(1a; S-isomer): ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.03 (br. S., 1 H) 7.78 (d, J = 7.05 Hz, 1 H) 7.61 (d, J = 7.55 Hz, 1 H) 7.27-7.37 (m, 1 H) 7.11-7.23 (m, 1 H) 3.89 (d, J = 10.07 Hz, 1 H) 3.64 (d, J = 10.07 Hz, 1 H) 2.96 -3.09 (m, 2 H) 2.71-2.88 (m, 2 H) 2.66 (t, J = 7.81 Hz, 2 H) 2.02-2.11 (m, 1 H) 1.85-1.97 (m, 1 H) 1.41-1.65 (m, 3 H). MS (LC / MS) RT = 1.15; [M + H] ⁺ = 315.3. Optical rotation (1.23 mg / mL, DMSO) = + 5.20 °.

The second peak is (R) -N- (benzo [d] thiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- It was an amine (1.21 g, 3.85 mmol, yield 24.5%).
(1b; R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.02 (br. S., 1 H) 7.79 (d, J = 7.32 Hz, 1 H) 7.62 (d, J = 7.63 Hz, 1 H) 7.33 (t, J = 7.63 Hz, 1 H) 7.18 (t, J = 7.48 Hz, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 2.98-3.10 (m, 2 H) 2.73-2.87 (m, 2 H) 2.67 (t, J = 7.63 Hz, 2 H) 2.08 (br. S., 1 H) 1.93 (br. S ., 1 H) 1.42-1.67 (m, 3 H). MS (LC / MS) RT = 1.15; [M + H] ⁺ = 315.3; Optical rotation (3.9 mg / mL, DMSO) = -3.92 °.

Example 2
N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

メカニカルスターラー、滴下漏斗および温度計を備えた５００mlの３ツ首フラスコに、酢酸(１００ml)を加え、氷浴中で冷却した。チオシアン酸カリウム(４０ｇ, ４１２mmol)および６−メトキシピリジン−３−アミン(６.２ｇ, ４９.９mmol)を反応混合物に加えた。反応物の温度が＜０℃に至るまで、反応物を氷−塩浴中で冷却した。反応温度を＜０℃に維持する速度で、酢酸(３０.０ml)中の臭素(８ml, １５６mmol)の溶液を２時間かけて滴下した。メカニカルスターラーでの撹拌を要した。添加完了後、混合物を撹拌しながら、ゆっくりと一夜かけて室温まで温めた。水(３０ml)を加え、混合物を、油浴中、８５℃まで加熱した。熱いままで、この混合物を濾過した。橙色のフィルターケーキを反応フラスコに戻し、さらに５０mlの酢酸を加えた。混合物を再度８５℃まで加熱し、熱いままで１回以上濾過した。合わせた濾液を氷浴中で冷却し、濃水酸化アンモニウムでｐＨ ８まで中和した。紫色の沈殿物が形成した。これを濾過によって集め、５ｇの粗製の物質を得た。この粗製の物質をメタノール(４０ml)から再結晶し、５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−アミンを紫色の結晶として得た(３ｇ, １６.５５mmol, 収率３３.１％)。
¹H NMR (300 MHz, DMSO-d₆) δ ppm 7.60 (1 H, d, J=8.42 Hz), 7.41 (2 H, s), 6.67 (1 H, d, J=8.78 Hz), 3.81 (3 H, s)。 Step A: 5-Methoxythiazolo [5,4-b] pyridin-2-amine

Acetic acid (100 ml) was added to a 500 ml three-necked flask equipped with a mechanical stirrer, dropping funnel and thermometer 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 temperature of the reaction reached <0 ° C. A solution of bromine (8 ml, 156 mmol) in acetic acid (30.0 ml) was added dropwise over 2 hours at a rate maintaining the reaction temperature <0 ° C. Stirring with a mechanical stirrer was required. After the addition was complete, the mixture was allowed to warm slowly to room temperature overnight with stirring. Water (30 ml) was added and the mixture was heated to 85 ° C. in an oil bath. The mixture was filtered until hot. The orange filter cake was returned to the reaction flask and an additional 50 ml of acetic acid was added. The mixture was heated again to 85 ° C. and filtered hot once more. The combined filtrate was cooled in an ice bath and neutralized to pH 8 with concentrated ammonium hydroxide. A purple precipitate formed. This was collected by filtration to give 5 g of crude material. This crude material was recrystallized from methanol (40 ml) to give 5-methoxythiazolo [5,4-b] pyridin-2-amine as purple crystals (3 g, 16.55 mmol, 33.1 yield). %).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 7.60 (1 H, d, J = 8.42 Hz), 7.41 (2 H, s), 6.67 (1 H, d, J = 8.78 Hz), 3.81 ( 3 H, s).

５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−アミン(２.４ｇ, １３.２４mmol)を、５×２０mlのネジ蓋付きバイアルに分けた。各バイアルに、アセトニトリル(１０ml)およびチオカルボニルジイミダゾール(６００mg)を加えた。全てのバイアルを６０℃で一夜加熱した。反応バイアルを合わせて、濃縮し、粗生成物のＮ−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た。この粗生成物を、Ｎ,Ｎ−ジメチルホルムアミド(５０ml)に懸濁し、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(２.７ｇ, １５.８８mmol)を加えた。反応物を７０℃で４時間加熱した。ＬＣ／ＭＳにより、本質的に変換の完了が示された。反応物を室温まで冷却し、水に注いだ。生成物を最初にトルエンで抽出し、次にクロロホルムで抽出した。有機物を合わせて、水で、そして塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、粗製の物質を得た。この粗製の物質をフラッシュクロマトグラフィーによって精製した(２０〜１００％ 酢酸エチル−ヘキサン)。生成物のフラクションを集めて、真空で濃縮し、(３−ヒドロキシ−３−((３−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを得た(１.３６ｇ, ３.４６mmol, 収率２６.１％)。^１Ｈ−ＮＭＲにより、(３−ヒドロキシ−３−((３−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.３６ｇ, ３.４６mmol, 収率２６.１％)と５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−アミン(.３４ｇ, １.８７６mmol, 収率１４.１７％)のモル比が１：０.５５であることが示された。混合物を、さらに精製することなく次の工程に直接用いた。
MS (LC/MS) R.T. = 3.23; [M+H]⁺ = 392.1。 Step B: (3-Hydroxy-3-((3- (5-methoxythiazolo [5,4-b] pyridin-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] Octane-1-yl) trihydroborate

5-Methoxythiazolo [5,4-b] pyridin-2-amine (2.4 g, 13.24 mmol) was divided into 5 × 20 ml screw cap vials. To each vial was added acetonitrile (10 ml) and thiocarbonyldiimidazole (600 mg). All vials were heated at 60 ° C. overnight. The reaction vials were combined and concentrated to give the crude product N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -1H-imidazole-1-carbothioamide. This crude product was suspended in N, N-dimethylformamide (50 ml) and (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) tri Hydroborate (2.7 g, 15.88 mmol) was added. The reaction was heated at 70 ° C. for 4 hours. LC / MS showed essentially complete conversion. The reaction was cooled to room temperature and poured into water. The product was first extracted 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 give the crude material. The crude material was purified by flash chromatography (20-100% ethyl acetate-hexane). The product fractions were collected and concentrated in vacuo to give (3-hydroxy-3-((3- (5-methoxythiazolo [5,4-b] pyridin-2-yl) thioureido) methyl) -1- Ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate was obtained (1.36 g, 3.46 mmol, yield 26.1%). ^{According to 1} H-NMR, (3-hydroxy-3-((3- (5-methoxythiazolo [5,4-b] pyridin-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2 .2] octane-1-yl) trihydroborate (1.36 g, 3.46 mmol, 26.1% yield) and 5-methoxythiazolo [5,4-b] pyridin-2-amine (.34 g, The molar ratio of 1.876 mmol (14.17% yield) was shown to be 1: 0.55. The mixture was used directly in the next step without further purification.
MS (LC / MS) RT = 3.23; [M + H] ⁺ = 392.1.

Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中の(３−ヒドロキシ−３−((３−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.７ｇ, ３.４６mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(１.８９ml, １２.１０mmol)を加えた。反応物を７０℃で２時間撹拌した。混合物を冷却し、水に注いだ。生成物をトルエンおよびクロロホルムで抽出した。有機層を合わせ、塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、粗製の物質を得た。この固体の物質をエーテルで磨砕した。固体を濾過して乾燥させ、(２−(５−メトキシ−３ａ,７ａ−ジヒドロチアゾロ[５,４−ｂ]ピリジン−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを得た(７００mg, １.９４mmol, 収率５６.０％)。
¹H NMR (300 MHz, DMSO-d₆) δ ppm 8.98 (1 H, s), 7.86 (1 H, d, J=8.78 Hz), 6.81 (1 H, d, J=8.42 Hz), 3.87 (3 H, s), 3.83 (1 H, s), 3.68 - 3.78 (1 H, m), 3.31 (1 H, s), 3.15 - 3.29 (1 H, m), 2.78 - 3.14 (4 H, m), 2.26 (1 H, br. s.), 2.04 (1 H, br. s.), 1.63 - 1.89 (3 H, m)。 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

(3-Hydroxy-3-((3- (5-methoxythiazolo [5,4-b] pyridin-2-yl) thioureido) methyl) -1-ammonio in N, N-dimethylformamide (10 ml) To bicyclo [2.2.2] octan-1-yl) trihydroborate (1.7 g, 3.46 mmol) was added N, N′-diisopropylcarbodiimide (1.89 ml, 12.10 mmol). The reaction was stirred at 70 ° C. for 2 hours. The mixture was cooled and 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 give the crude material. This solid material was triturated with ether. The solid was filtered and dried to give (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 was obtained (700 mg, 1.94 mmol, 56.0% yield).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 8.98 (1 H, s), 7.86 (1 H, d, J = 8.78 Hz), 6.81 (1 H, d, J = 8.42 Hz), 3.87 ( 3 H, s), 3.83 (1 H, s), 3.68-3.78 (1 H, m), 3.31 (1 H, s), 3.15-3.29 (1 H, m), 2.78-3.14 (4 H, m ), 2.26 (1 H, br. S.), 2.04 (1 H, br. S.), 1.63-1.89 (3 H, m).

アセトン(１０ml)中の(２−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(７８０mg, ２.１７mmol)に、３Ｍ ＨＣｌ(１０ml, ３２９mmol)を加えた。反応物を室温で２時間撹拌した。クロロホルムおよび水を加え、水層を分離した。水層を重炭酸ナトリウムで中和した。生成物をクロロホルム(２×)で抽出した。有機層を合わせ、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(５−メトキシチアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(５８８mg, １.７０mmol, 収率７８％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.90 (1 H, br. s.), 7.82 - 7.86 (1 H, m), 6.80 (1 H, d, J=8.85 Hz), 3.84 - 3.89 (4 H, m), 3.61 (1 H, d, J=10.07 Hz), 3.03 (2 H, d, J=5.19 Hz), 2.73 - 2.86 (2 H, m), 2.66 (2 H, t, J=7.78 Hz), 2.07 (1 H, br. s.), 1.92 (1 H, br. s.), 1.45 - 1.65 (3 H, m). MS (LC/MS) R.T. = 1.29; [M+H]⁺ = 346.1。 Step D: N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane]- 2-Amine

(2- (5-Methoxythiazolo [5,4-b] pyridin-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2] in acetone (10 ml). .2] octane] -1′-yl) trihydroborate (780 mg, 2.17 mmol) 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 separated. The aqueous layer was neutralized with sodium bicarbonate. The product was extracted with chloroform (2x). The organic layers were combined, dried over sodium sulfate, filtered, concentrated in vacuo, and racemic N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [ Oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (588 mg, 1.70 mmol, 78% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.90 (1 H, br. S.), 7.82-7.86 (1 H, m), 6.80 (1 H, d, J = 8.85 Hz), 3.84- 3.89 (4 H, m), 3.61 (1 H, d, J = 10.07 Hz), 3.03 (2 H, d, J = 5.19 Hz), 2.73-2.86 (2 H, m), 2.66 (2 H, t , J = 7.78 Hz), 2.07 (1 H, br. S.), 1.92 (1 H, br. S.), 1.45-1.65 (3 H, m). MS (LC / MS) RT = 1.29; [ M + H] ⁺ = 346.1.

The enantiomers were separated on a mobile phase consisting of 35% methanol in CO ₂ (0.1% DEA) using a Chiralpak AD-H (30 × 250 mm, 5 μm) column. The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2 2.2.2 Octane] -2-amine (212 mg, 0.61 mmol, 36.1% yield).
(2a; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.89 (1 H, br. S.), 7.84 (1 H, d, J = 8.85 Hz), 6.77-6.82 (1 H, m), 3.84-3.89 (4 H, m), 3.61 (1 H, d, J = 10.07 Hz), 3.03 (2 H, d, J = 5.19 Hz), 2.74-2.86 (2 H, m), 2.66 (2 H, t, J = 7.63 Hz), 2.06 (1 H, br. s.), 1.92 (1 H, br. s.), 1.44-1.65 (3 H, m). MS ( LC / MS) RT = 1.47; [M + H] ⁺ = 346.2. Optical rotation (3.57 mg / ml, DMSO) = -2.58 °.

The second peak is (R) -N- (5-methoxythiazolo [5,4-b] pyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2. 2.2] octane] -2-amine (242 mg, 0.70 mmol, 41.2% yield).
(2b; R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.89 (1 H, br. S.), 7.84 (1 H, d, J = 8.85 Hz), 6.79 (1 H, d, J = 8.55 Hz), 3.82-3.90 (4 H, m), 3.61 (1 H, d, J = 10.07 Hz), 3.03 (2 H, d, J = 5.49 Hz), 2.74-2.86 ( 2 H, m), 2.66 (2 H, t, J = 7.78 Hz), 2.06 (1 H, br. S.), 1.92 (1 H, br. S.), 1.42-1.67 (3 H, m) MS (LC / MS) RT = 1.30; [M + H] ⁺ = 346.2. Optical rotation (3.29 mg / ml, DMSO) = + 2.43 °.

Example 3
(2- (5H-1′-azaspiro [oxazol-4,3′-bicyclo [2.2.2] octane] -2-ylamine) benzo [d] thiazol-6-yl) pyrrolidin-1-yl) methanone

２５０mlのフラスコに、テトラヒドロフラン(５０ml)中の２−(ｔｅｒｔ−ブトキシカルボニル−アミノ)ベンゾ[ｄ]チアゾール−６−カルボン酸(１.０ｇ, ３.４mmol)およびピロリジン(０.５５９ml, ６.８mmol)を加えた。この溶液に、ＥＤＣ(１.３ｇ, ６.８mmol)、１−ヒドロキシベンゾトリアゾール(triaxole)(１.０４１ｇ, ６.８mmol)およびヒューニッヒ塩基(２.３７ml, １３.５９mmol)を加えた。反応物を２５℃で１時間撹拌した。反応物を水およびジクロロメタンに注いだ。その水をジクロロメタンで３回抽出し、有機層を合わせ、濃縮した。残渣を少量のジクロロメタンに溶かし、ジエチルエーテル／ヘキサンで沈殿させた。フラスコを冷蔵庫中に１時間置いて、濾過した。白色の沈殿物を集め、６−(ピロリジン−１−カルボニル)ベンゾ[ｄ]チアゾール−２−イルカルバミン酸ｔｅｒｔ−ブチルを得た(１.０９ｇ, ３.１４mmol, 収率９２％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.95 (s, 1 H), 8.14 (s, 1 H), 7.61 - 7.77 (m, J=8.39, 1.98 Hz, 1 H), 7.46 - 7.63 (m, 1 H), 3.39 - 3.63 (m, 4 H), 1.74 - 2.00 (m, 4 H), 1.45 - 1.62 (m, 9 H). MS (LC/MS) R.T. = 3.40; [M+H]⁺ = 363.1。 Step A: tert-Butyl 6- (pyrrolidin-1-carbonyl) benzo [d] thiazol-2-ylcarbamate

A 250 ml flask was charged with 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). ) Was added. To this solution was added EDC (1.3 g, 6.8 mmol), 1-hydroxybenzotriazole (1.041 g, 6.8 mmol) and Hunig's base (2.37 ml, 13.59 mmol). The reaction was stirred at 25 ° C. for 1 hour. The reaction was poured into water and dichloromethane. The water was extracted 3 times with dichloromethane and the organic layers were combined and concentrated. The residue was dissolved in a small amount of dichloromethane and precipitated with diethyl ether / hexane. The flask was placed in the refrigerator for 1 hour and filtered. The white precipitate was collected to give tert-butyl 6- (pyrrolidin-1-carbonyl) benzo [d] thiazol-2-ylcarbamate (1.09 g, 3.14 mmol, 92% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.95 (s, 1 H), 8.14 (s, 1 H), 7.61-7.77 (m, J = 8.39, 1.98 Hz, 1 H), 7.46-7.63 (m, 1 H), 3.39-3.63 (m, 4 H), 1.74-2.00 (m, 4 H), 1.45-1.62 (m, 9 H). MS (LC / MS) RT = 3.40; (M + H] ⁺ = 363.1.

６−(ピロリジン−１−カルボニル)ベンゾ[ｄ]チアゾール−２−イルカルバミン酸ｔｅｒｔ−ブチル(１.０９ｇ, ３.１４mmol)を、ジクロロメタン(１０ml)およびＴＦＡ(３ml, ３８.９mmol)に溶解し、反応物を２５℃で一夜撹拌した。反応物を分液漏斗に注ぎ、重炭酸ナトリウムで注意深く中和した。液体をクロロホルム／メタノール(４：１)で３回抽出した。有機層を濃縮し、白色の残渣を得て、ジエチルエーテルで磨砕した。沈殿物を集め、(２−アミノベンゾ[ｄ]チアゾール−６−イル)(ピロリジン−１−イル)メタノンを得た(０.４９７ｇ, ２.０mmol, ６４％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.86 (d, J=1.51 Hz, 1 H), 7.65 (s, 2 H), 7.35 - 7.46 (m, 1 H), 7.26 - 7.35 (m, 1 H), 3.41 - 3.57 (m, 4 H), 1.64 - 1.97 (m, 4 H). MS (LC/MS) R.T. = 1.39; [M+H]⁺ = 248.1。 Step B: (2-Aminobenzo [d] thiazol-6-yl) (pyrrolidin-1-yl) methanone

Tert-butyl 6- (pyrrolidin-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). The reaction was stirred at 25 ° 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 layer was concentrated to give a white residue that was triturated with diethyl ether. The precipitate was collected to give (2-aminobenzo [d] thiazol-6-yl) (pyrrolidin-1-yl) methanone (0.497 g, 2.0 mmol, 64%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.86 (d, J = 1.51 Hz, 1 H), 7.65 (s, 2 H), 7.35-7.46 (m, 1 H), 7.26-7.35 (m , 1 H), 3.41-3.57 (m, 4 H), 1.64-1.97 (m, 4 H). MS (LC / MS) RT = 1.39; [M + H] ⁺ = 248.1.

(２−(５Ｈ−１'−アザスピロ[オキサゾール−４,３'−ビシクロ[２.２.２]オクタン]−２−イルアミン)ベンゾ[ｄ]チアゾール−６−イル)ピロリジン−１−イル)メタノンを、実施例１の工程Ａ〜Ｄの一般的な手順に従って、(２−アミノベンゾ[ｄ]チアゾール−６−イル)(ピロリジン−１−イル)メタノン(実施例３の工程Ｂ)を出発物質として用いて製造した。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.05 (br. s., 1 H) 7.99 (d, J=1.76 Hz, 1 H) 7.60 (d, J=8.31 Hz, 1 H) 7.48 (dd, J=8.31, 1.76 Hz, 1 H) 3.90 (d, J=10.07 Hz, 1 H) 3.65 (d, J=10.07 Hz, 1 H) 3.42 - 3.52 (m, 4 H) 3.04 (s, 2 H) 2.75 - 2.87 (m, 2 H) 2.67 (t, J=7.81 Hz, 2 H) 2.08 (br. s., 1 H) 1.76 - 1.98 (m, 5 H) 1.41 - 1.65 (m, 3 H). MS (LC/MS) R.T. = 1.33; [M+H]⁺ = 412.2。 Step C: (2- (5H-1′-azaspiro [oxazole-4,3′-bicyclo [2.2.2] octane] -2-ylamine) benzo [d] thiazol-6-yl) pyrrolidin-1- Il) methanone

(2- (5H-1′-azaspiro [oxazol-4,3′-bicyclo [2.2.2] octane] -2-ylamine) benzo [d] thiazol-6-yl) pyrrolidin-1-yl) methanone According to the general procedure of steps AD of Example 1, starting from (2-aminobenzo [d] thiazol-6-yl) (pyrrolidin-1-yl) methanone (Step B of Example 3) Manufactured.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.05 (br.s., 1 H) 7.99 (d, J = 1.76 Hz, 1 H) 7.60 (d, J = 8.31 Hz, 1 H) 7.48 ( dd, J = 8.31, 1.76 Hz, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 3.42-3.52 (m, 4 H) 3.04 (s, 2 H) 2.75-2.87 (m, 2 H) 2.67 (t, J = 7.81 Hz, 2 H) 2.08 (br. S., 1 H) 1.76-1.98 (m, 5 H) 1.41-1.65 (m, 3 H ). MS (LC / MS) RT = 1.33; [M + H] ⁺ = 412.2.

Example 4
N- (5-phenylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(６ml)中の５−フェニルチアゾール−２−アミン(０.５２ｇ, ２.９mmol)に、１,１'−チオカルボニルジイミダゾール(０.６８ｇ, ３.８mmol)を加えた。反応混合物を６５℃で２時間撹拌した。沈殿物を濾過し、アセトニトリル(２×２０ml)で洗浄し、中間体Ｎ−(５−フェニルチアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た。中間体をＮ,Ｎ−ジメチルホルムアミド(３０ml)に溶かし、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５ｇ, ２.９mmol)で処理した。反応混合物を６５℃で５時間撹拌した。反応物を真空で濃縮し、シリカゲルのクロマトグラフィーによって精製によって精製した(３０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを合わせて、真空で濃縮し、(３−ヒドロキシ−３−((３−(５−フェニルチアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(.８５ｇ, ２.１９mmol, 収率７４.４％)。ＬＣ／ＭＳにより、本ＬＣ／ＭＳ条件でＢＨ_３が無い生成物を確認した：保持時間 3.26 (M+1-BH₃= 375.33)。 Step A: (3-Hydroxy-3-((3- (5-phenylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate

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 ° C. for 2 hours. The precipitate was filtered and washed with acetonitrile (2 × 20 ml) to give the intermediate N- (5-phenylthiazol-2-yl) -1H-imidazole-1-carbothioamide. The intermediate was dissolved in N, N-dimethylformamide (30 ml) and (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate (0 0.5 g, 2.9 mmol). The reaction mixture was stirred at 65 ° C. for 5 hours. The reaction was concentrated in vacuo and purified by purification on silica gel (30-100% ethyl acetate / hexanes). The product fractions were combined and concentrated in vacuo to give (3-hydroxy-3-((3- (5-phenylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2. ] Octan-1-yl) trihydroborate was obtained as a white powder (0.85 g, 2.19 mmol, 74.4% yield). LC / MS confirmed the product without BH _{3 under the} present LC / MS conditions: retention time 3.26 (M + 1-BH ₃ = 375.33).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−ヒドロキシ−３−((３−(５−フェニルチアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.８ｇ, ２.１mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(１.１２ml, ７.２mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を真空で濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。合わせた生成物のフラクションを真空で濃縮し、(２−(５−フェニルチアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.５１ｇ, １.４４mmol, 収率７０％)。ＬＣＭＳ−マスは、ＬＣ／ＭＳ条件でＢＨ_３が無いものに対応する：保持時間 2.46 (M+1-BH₃ = 341.36)。 Step B: (2- (5-Phenylthiazol-2-ylamino) -4H-1′-ammoniospiro- [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) Trihydroborate

(3-Hydroxy-3-((3- (5-phenylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] in N, N-dimethylformamide (20 ml). To octan-1-yl) trihydroborate (0.8 g, 2.1 mmol) was added N, N′-diisopropylcarbodiimide (1.12 ml, 7.2 mmol). The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated in vacuo and purified by chromatography on silica gel (40-100% ethyl acetate / hexane). The combined product fractions were concentrated in vacuo to give (2- (5-phenylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2]. Octane] -1′-yl) trihydroborate was obtained as a white powder (0.51 g, 1.44 mmol, 70% yield). LCMS- mass corresponds to that no BH ₃ by LC / MS conditions: retention time _{2.46 (M + 1-BH 3} = 341.36).

アセトン(９ml)中の(２−(５−フェニルチアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.５６ｇ, １.５８mmol)に、３Ｍ ＨＣｌ(３.９５ml, １１.８６mmol)を加えた。反応混合物を室温で４時間撹拌し、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×２０ml)で、次にクロロホルム(２×２０ml)で抽出した。有機層を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、白色の粉末を得た。粗生成物を逆相ＨＰＬＣによって精製し(Phenomenex Luna 30×100mm；波長 ２２０nm；濃度勾配 １０分；流速 ４０ml/分；溶媒Ａ＝１０％ メタノール−９０％ 水−０.１％ ＴＦＡ、溶媒Ｂ＝９０％ メタノール−１０％ 水−０.１％ ＴＦＡ)。フラクションを合わせて、１Ｎ 水酸化ナトリウムで中和し、酢酸エチル(２×３０ml)およびクロロホルム(２×３０ml)で抽出した。有機物を合わせ、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(５−フェニルチアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.４ｇ, １.１７５mmol, 収率７４.３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.63 (1 H, br. s.), 7.71 (1 H, s), 7.52 (2 H, d, J=7.32 Hz), 7.37 (2 H, t, J=7.78 Hz), 7.25 (1 H, t, J=7.32 Hz), 3.82 (1 H, d, J=10.07 Hz), 3.57 (1 H, d, J=9.77 Hz), 3.02 (2 H, d, J=4.27 Hz), 2.79 (2 H, t, J=7.63 Hz), 2.66 (2 H, t, J=7.63 Hz), 2.04 (1 H, br. s.), 1.92 - 1.97 (1 H, m), 1.44 - 1.65 (3 H, m). MS (LC/MS) R.T. = 1.52; [M+H]⁺ = 341.3。 Step C: N- (5-Phenylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (5-Phenylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- in acetone (9 ml) Yl) trihydroborate (0.56 g, 1.58 mmol) was added 3M HCl (3.95 ml, 11.86 mmol). The reaction mixture was stirred at room temperature for 4 hours and neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 20 ml) and then with chloroform (2 × 20 ml). The organic layers were combined, dried over magnesium sulfate, filtered and concentrated in vacuo to give a white powder. The crude product was purified by reverse phase HPLC (Phenomenex Luna 30 × 100 mm; wavelength 220 nm; concentration gradient 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). Fractions were combined, neutralized with 1N sodium hydroxide and extracted with ethyl acetate (2 × 30 ml) and chloroform (2 × 30 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (5-phenylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2. .2] Octane] -2-amine was obtained as a white powder (0.4 g, 1.175 mmol, 74.3% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.63 (1 H, br. S.), 7.71 (1 H, s), 7.52 (2 H, d, J = 7.32 Hz), 7.37 (2 H , t, J = 7.78 Hz), 7.25 (1 H, t, J = 7.32 Hz), 3.82 (1 H, d, J = 10.07 Hz), 3.57 (1 H, d, J = 9.77 Hz), 3.02 ( 2 H, d, J = 4.27 Hz), 2.79 (2 H, t, J = 7.63 Hz), 2.66 (2 H, t, J = 7.63 Hz), 2.04 (1 H, br.s.), 1.92- 1.97 (1 H, m), 1.44-1.65 (3 H, m). MS (LC / MS) RT = 1.52; [M + H] ⁺ = 341.3.

The enantiomers were separated using a Chiralpak AS-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 300 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.4 g, 1.15 mmol, 32.7% were obtained.
(4a, S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.69 (s, 1 H), 7.69-7.87 (m, 1 H), 7.52-7.66 (m, J = 10.99 Hz, 2 H), 7.37-7.51 (m, 2 H), 7.21-7.38 (m, 2 H), 3.52-4.00 (m, 2 H), 2.98-3.23 (m, 2 H), 2.78-2.94 ( m, 2 H), 2.64-2.78 (m, 2 H), 1.88-2.19 (m, J = 60.43 Hz, 2 H), 1.40-1.77 (m, 3 H). MS (LC / MS) RT = 1.77 ; [M + H] ⁺ = 341.1.

From the second peak, 0.4 g, 1.15 mmol, 32.7% were obtained.
(4b, R-isomer): L melting point 187-9 ° C. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.64 (s, 1 H), 7.62-7.84 (m, 1 H), 7.44- 7.61 (m, 2 H), 7.33-7.46 (m, 2 H), 7.18-7.31 (m, 1 H), 3.50-3.99 (m, 2 H), 2.94-3.14 (m, 2 H), 2.74- 2.91 (m, 2 H), 2.61-2.72 (m, 2 H), 2.05 (s, 1 H), 1.82-2.00 (m, 1 H), 1.34-1.72 (m, 3 H). MS (LC / MS) RT = 1.78; [M + H] ⁺ = 341.1.

アセトニトリル(２０ml)中の６−メトキシベンゾ[ｄ]チアゾール−２−アミン(０.５３ｇ, ２.９４mmol)に、１,１'−チオカルボニルジイミダゾール(０.６８１ｇ, ３.８２mmol)を加えた。反応混合物を６５℃で２４時間撹拌した。沈殿物を濾過し、アセトニトリル(２×２０ml)で洗浄し、生成物を得た。生成物を、さらに精製せず、または特性決定せずに、次の工程に直接用いた。 Example 5
N- (6-methoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine
Step A: N- (6-Methoxybenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

To 1-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 ° C. for 24 hours. The precipitate was filtered and washed with acetonitrile (2 × 20 ml) to give the product. The product was used directly in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(６−メトキシベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(.８２ｇ, ２.８２mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.４８ｇ, ２.８２mmol)を加えた。反応混合物を６５℃で６時間撹拌した。反応物を真空で濃縮し、シリカゲルのクロマトグラフィーによって精製した(３０％〜１００％ 酢酸エチル／ヘキサン)。純粋なフラクションを合わせて、濃縮し、(３−ヒドロキシ−３−((３−(６−メトキシベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.７ｇ, １.７８mmol, 収率６３.２％)。ＬＣ／ＭＳにより、本ＬＣ／ＭＳ条件でＢＨ_３が無い生成物を確認した：保持時間 3.11 (M+1-BH₃ = 379.4)。 Step B: (3-Hydroxy-3-((3- (6-methoxybenzo [d] thiazol-2-yl) -thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1- Yl) trihydroborate

N- (6-methoxybenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.82 g, 2.82 mmol) in N, N-dimethylformamide (20 ml) was added to (3- ( Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.48 g, 2.82 mmol) was added. The reaction mixture was stirred at 65 ° C. for 6 hours. The reaction was concentrated in vacuo and purified by silica gel chromatography (30-100% ethyl acetate / hexanes). The pure fractions were combined and concentrated to give (3-hydroxy-3-((3- (6-methoxybenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2. 2] Octane-1-yl) trihydroborate was obtained as a white powder (0.7 g, 1.78 mmol, 63.2% yield). LC / MS confirmed the product without BH _{3 under the} present LC / MS conditions: retention time 3.11 (M + 1-BH ₃ = 379.4).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−ヒドロキシ−３−((３−(６−メトキシベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.６８ｇ, １.７３mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.９５ml, ６.１mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを合わせて、真空で濃縮し、(２−(６−メトキシベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.３５ｇ, ０.９８mmol, 収率５６.４％)。
LC/MS MH⁺ - BH₃ = 345.2。 Step C: (2- (6-Methoxybenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1 ′ -Yl) trihydroborate

(3-Hydroxy-3-((3- (6-methoxybenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2 in N, N-dimethylformamide (20 ml) .2] To octan-1-yl) trihydroborate (0.68 g, 1.73 mmol) was added N, N′-diisopropylcarbodiimide (0.95 ml, 6.1 mmol). The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were combined and concentrated in vacuo to give (2- (6-methoxybenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2]. 2.2.2] Octane] -1′-yl) trihydroborate was obtained as a white powder (0.35 g, 0.98 mmol, 56.4% yield).
_{^{LC / MS MH + - BH 3}} = 345.2.

アセトン(９ml)中の(２−(６−メトキシベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.３３ｇ, ０.９２１mmol)に、３Ｍ ＨＣｌ(２.３０ml, ６.９１mmol)を加えた。反応物を室温で４時間撹拌し、１Ｎ 水酸化ナトリウムで中和した。生成物を、酢酸エチル(２×２０ml)で、次にクロロホルム(２×２０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、白色の粉末を得た。粗生成物を逆相ＨＰＬＣによって精製した(Phenomenex Luna 30×100mm；波長 ２２０nm；濃度勾配 １０分；流速 ４０ml/分；溶媒Ａ＝１０％ メタノール−９０％ 水−０.１％ ＴＦＡ、溶媒Ｂ＝９０％ メタノール−１０％ 水−０.１％ ＴＦＡ)。フラクションを合わせて、１Ｎ 水酸化ナトリウムで中和し、酢酸エチル(２×３０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(６−メトキシベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.２５ｇ, ０.７３mmol, 収率７９％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.88 (1 H, d, J=1.22 Hz), 7.48 - 7.52 (1 H, m), 7.40 (1 H, d, J=2.75 Hz), 6.92 (1 H, dd, J=8.70, 2.59 Hz), 3.87 (1 H, d, J=9.77 Hz), 3.77 (3 H, s), 3.61 (1 H, d, J=9.77 Hz), 3.03 (2 H, s), 2.75 - 2.86 (2 H, m), 2.67 (2 H, t, J=7.78 Hz), 2.06 (1 H, br. s.), 1.91 (1 H, br. s.), 1.41 - 1.65 (3 H, m). MS (LC/MS) R.T. = 1.44; [M+H]⁺ = 345.3。 Step D: N- (6-Methoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (6-Methoxybenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] in acetone (9 ml) To 1'-yl) trihydroborate (0.33 g, 0.921 mmol) was added 3M HCl (2.30 ml, 6.91 mmol). The reaction was stirred at room temperature for 4 hours and neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 20 ml) and then with chloroform (2 × 20 ml). The organics were combined, dried over magnesium sulfate, filtered and concentrated in vacuo to give a white powder. The crude product was purified by reverse phase HPLC (Phenomenex Luna 30 × 100 mm; wavelength 220 nm; concentration gradient 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). Fractions were combined, neutralized with 1N sodium hydroxide and extracted with ethyl acetate (2 × 30 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (6-methoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′- Bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.25 g, 0.73 mmol, 79% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.88 (1 H, d, J = 1.22 Hz), 7.48-7.52 (1 H, m), 7.40 (1 H, d, J = 2.75 Hz), 6.92 (1 H, dd, J = 8.70, 2.59 Hz), 3.87 (1 H, d, J = 9.77 Hz), 3.77 (3 H, s), 3.61 (1 H, d, J = 9.77 Hz), 3.03 (2 H, s), 2.75-2.86 (2 H, m), 2.67 (2 H, t, J = 7.78 Hz), 2.06 (1 H, br.s.), 1.91 (1 H, br. S. ), 1.41-1.65 (3 H, m). MS (LC / MS) RT = 1.44; [M + H] ⁺ = 345.3.

The enantiomers were separated using a Chiralpak AD-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 23% methanol in CO ₂ (0.1% DEA) while monitoring UV at 220 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 205.5 mg, 0.60 mmol, 34.1% were obtained.
(5a, S-isomer): ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.88 (1 H, br. S.), 7.50 (1 H, d, J = 8.81 Hz), 7.40 (1 H, d, J = 2.52 Hz), 6.92 (1 H, dd, J = 8.81, 2.77 Hz), 3.87 (1 H, d, J = 9.82 Hz), 3.77 (3 H, s), 3.58-3.65 ( 1 H, m), 3.02 (2 H, s), 2.74-2.86 (2 H, m), 2.66 (2 H, t, J = 7.68 Hz), 2.03-2.08 (1 H, m), 1.91 (1 H, br. S.), 1.39-1.65 (3 H, m). MS (LC / MS) RT = 1.40; [M + H] ⁺ = 345.2.

From the second peak, 206.9 mg, 0.6 mmol, 34% were obtained.
(5b, R-isomer): ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.88 (1 H, br. S.), 7.50 (1 H, d, J = 8.56 Hz), 7.40 (1 H, d, J = 2.52 Hz), 6.93 (1 H, dd, J = 8.81, 2.52 Hz), 3.87 (1 H, d, J = 10.07 Hz), 3.77 (3 H, s), 3.62 (1 H , d, J = 10.07 Hz), 3.02 (2 H, s), 2.75-2.86 (2 H, m), 2.67 (2 H, t, J = 7.68 Hz), 2.04-2.09 (1 H, m), 1.92 (1 H, br. S.), 1.42-1.66 (3 H, m). MS (LC / MS) RT = 1.70; [M + H] ⁺ = 345.1.

Example 6
N- (4-Methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の４−メチルベンゾ[ｄ]チアゾール−２−アミン(１.１ｇ, ６.７mmol)に、１,１'−チオカルボニルジイミダゾール(１.５５２ｇ, ８.７１mmol)を加えた。反応混合物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(５０℃)中で１時間乾燥させ、Ｎ−(４−メチルベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(９００mg, ３.２８mmol, 収率４９％)を得た。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (4-Methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

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 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (50 ° C.) for 1 hour and N- (4-methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (900 mg, 3.28 mmol, yield) 49%). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(４−メチルベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.７１ｇ, ２.５９mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.４４ｇ, ２.５９mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−ヒドロキシ−３−((３−(４−メチルベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.６５ｇ, １.７３mmol, 収率６６.７％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.93 (1 H, s), 7.74 (1 H, d, J=7.63 Hz), 7.22 - 7.26 (1 H, m), 7.19 (1 H, t, J=7.63 Hz), 3.75 - 3.93 (2 H, m), 2.72 - 2.95 (6 H, m), 2.56 (3 H, s), 2.03 - 2.14 (1 H, m), 1.95 (1 H, br. s.), 1.78 - 1.87 (1 H, m), 1.73 (1 H, ddd, J=13.81, 9.23, 5.04 Hz), 1.56 (1 H, td, J=9.99, 7.78 Hz), 1.38 (2 H, br. s.). (LC/MS) R.T. = 3.70; [M+H]⁺ = 375.2。 Step B: (3-Hydroxy-3-((3- (4-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) Trihydroborate

N- (4-Methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.71 g, 2.59 mmol) in N, N-dimethylformamide (20 ml) was added to (3- ( Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.44 g, 2.59 mmol) was added. The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (3-hydroxy-3-((3- (4-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2. ] Octan-1-yl) trihydroborate was obtained as a white powder (0.65 g, 1.73 mmol, 66.7% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.93 (1 H, s), 7.74 (1 H, d, J = 7.63 Hz), 7.22-7.26 (1 H, m), 7.19 (1 H, t, J = 7.63 Hz), 3.75-3.93 (2 H, m), 2.72-2.95 (6 H, m), 2.56 (3 H, s), 2.03-2.14 (1 H, m), 1.95 (1 H , br. s.), 1.78-1.87 (1 H, m), 1.73 (1 H, ddd, J = 13.81, 9.23, 5.04 Hz), 1.56 (1 H, td, J = 9.99, 7.78 Hz), 1.38 (2 H, br. S.). (LC / MS) RT = 3.70; [M + H] ⁺ = 375.2.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−ヒドロキシ−３−((３−(４−メチルベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.６２ｇ, １.６５mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.３３ml, ２.１４mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(２−(４−メチルベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.４ｇ, １.１７mmol, 収率７０.９％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.90 (s, 1 H), 7.61 (d, J=7.63 Hz, 1 H), 7.13 - 7.17 (m, 1 H), 7.09 (t, J=7.48 Hz, 1 H), 3.90 (d, J=10.38 Hz, 1 H), 3.77 (d, J=10.38 Hz, 1 H), 3.32 (s, 3 H), 3.30 (d, J=1.53 Hz, 1 H), 3.13 - 3.20 (m, 2 H), 3.00 - 3.09 (m, 1 H), 2.85 - 2.94 (m, 4 H), 2.57 (s, 4 H), 2.28 (s, 1 H), 2.06 (s, 1 H), 1.75 - 1.83 (m, 4 H), 1.45 (s, 1 H). (LC/MS) R.T. = 2.73; [M+H]⁺ = 343.2。 Step C: (2- (4-Methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammonio-spiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- Yl) trihydroborate

(3-Hydroxy-3-((3- (4-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.] In N, N-dimethylformamide (20 ml). 2] To octan-1-yl) trihydroborate (0.62 g, 1.65 mmol) was added N, N'-diisopropylcarbodiimide (0.33 ml, 2.14 mmol). The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (2- (4-methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2]. ] Octane] -1'-yl) trihydroborate was obtained as a white powder (0.4 g, 1.17 mmol, yield 70.9%).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.90 (s, 1 H), 7.61 (d, J = 7.63 Hz, 1 H), 7.13-7.17 (m, 1 H), 7.09 (t, J = 7.48 Hz, 1 H), 3.90 (d, J = 10.38 Hz, 1 H), 3.77 (d, J = 10.38 Hz, 1 H), 3.32 (s, 3 H), 3.30 (d, J = 1.53 Hz , 1 H), 3.13-3.20 (m, 2 H), 3.00-3.09 (m, 1 H), 2.85-2.94 (m, 4 H), 2.57 (s, 4 H), 2.28 (s, 1 H) , 2.06 (s, 1 H), 1.75-1.83 (m, 4 H), 1.45 (s, 1 H). (LC / MS) RT = 2.73; [M + H] ⁺ = 343.2.

アセトン(９ml)中の(２−(４−メチルベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.３８ｇ, １.１１mmol)を３Ｍ ＨＣｌ(２.７８ml, ８.３３mmol)を加えた。反応混合物を室温で４時間撹拌した。酢酸エチルを加え、水層を集め、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(４−メチルベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.１９５ｇ, ０.５９４mmol, 収率５３.５％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.84 (s, 1 H), 7.60 (d, J=7.32 Hz, 1 H), 7.13 - 7.17 (m, 1 H), 7.08 (t, J=7.63 Hz, 1 H), 3.92 (d, J=10.07 Hz, 1 H), 3.66 (d, J=9.77 Hz, 1 H), 3.04 (s, 2 H), 2.76 - 2.85 (m, 2 H), 2.68 (t, J=7.48 Hz, 2 H), 2.56 (s, 3 H), 2.09 (s, 1 H), 1.93 (s, 1 H), 1.61 (d, J=3.05 Hz, 1 H), 1.60 (s, 1 H), 1.50 (dd, J=7.17, 2.59 Hz, 1 H). (LC/MS) R.T. = 1.76; [M+H]⁺ = 329.2。 Step D: N- (4-Methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (4-Methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane]-in acetone (9 ml) 1′-yl) trihydroborate (0.38 g, 1.11 mmol) was added to 3M 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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (4-methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.195 g, 0.594 mmol, 53.5% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.84 (s, 1 H), 7.60 (d, J = 7.32 Hz, 1 H), 7.13-7.17 (m, 1 H), 7.08 (t, J = 7.63 Hz, 1 H), 3.92 (d, J = 10.07 Hz, 1 H), 3.66 (d, J = 9.77 Hz, 1 H), 3.04 (s, 2 H), 2.76-2.85 (m, 2 H ), 2.68 (t, J = 7.48 Hz, 2 H), 2.56 (s, 3 H), 2.09 (s, 1 H), 1.93 (s, 1 H), 1.61 (d, J = 3.05 Hz, 1 H ), 1.60 (s, 1 H), 1.50 (dd, J = 7.17, 2.59 Hz, 1 H). (LC / MS) RT = 1.76; [M + H] ⁺ = 329.2.

The enantiomers were separated using a Chiralcel OJ-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 300 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.07 g, 0.21 mmol, 38.9% was obtained.
(6a, S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.83 (br. S., 1 H) 7.59 (d, J = 7.63 Hz, 1 H) 7.11-7.18 (m , 1 H) 7.08 (t, J = 7.63 Hz, 1 H) 3.92 (d, J = 10.38 Hz, 1 H) 3.66 (d, J = 9.77 Hz, 1 H) 3.04 (s, 2 H) 2.73-2.89 (m, 2 H) 2.61-2.72 (m, 2 H) 2.56 (s, 3 H) 2.09 (br. s., 1 H) 1.93 (br. s., 1 H) 1.43-1.71 (m, 3 H ). MS (LC / MS) RT = 1.75; [M + H] ⁺ = 329.1.

From the second peak, 0.07 g, 0.21 mmol, 38.1% were obtained.
(6b, R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.84 (br. S., 1 H) 7.59 (d, J = 7.63 Hz, 1 H) 7.15 (d, J = 7.20 Hz, 1 H) 7.08 (t, J = 7.48 Hz, 1 H) 3.92 (d, J = 9.77 Hz, 1 H) 3.66 (d, J = 10.07 Hz, 1 H) 3.00-3.09 (m, 2 H) 2.73-2.87 (m, 2 H) 2.62-2.72 (m, 2 H) 2.56 (s, 3 H) 2.05-2.12 (m, 1 H) 1.93 (br. S., 1 H) 1.56-1.67 ( m, 2 H) 1.45-1.55 (m, 1 H). MS (LC / MS) RT = 1.75; [M + H] ⁺ = 329.1.

Example 7
N- (4-Chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の４−クロロベンゾ[ｄ]チアゾール−２−アミン(１.１２ｇ, ６.０７mmol)に、１,１'−チオカルボニルジイミダゾール(１.４０５ｇ, ７.８９mmol)を加えた。反応混合物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を、真空オーブン(４０℃)中で１時間乾燥させ、Ｎ−(４−クロロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た(０.３３ｇ, １.１２mmol, 収率１８.５％)。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (4-Chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

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 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 1 hour to give N- (4-chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.33 g, 1.12 mmol, yield 18.5%). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(４−クロロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.３ｇ, １.０１８mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.１７３ｇ, １.０１８mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(６０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−((３−(４−クロロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.２５ｇ, ０.６３mmol, 収率６１.９％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 12.19 (s, 1 H) 9.72 (br. s., 1 H) 7.93 (d, J=7.93 Hz, 1 H) 7.51 (d, J=7.93 Hz, 1 H) 7.26 - 7.32 (m, 1 H) 5.32 (s, 1 H) 3.88 (dd, J=13.73, 4.88 Hz, 1 H) 3.75 (dd, J=13.73, 4.88 Hz, 1 H) 2.73 - 2.95 (m, 6 H) 2.08 (br. s., 1 H) 1.96 (br. s., 1 H) 1.79 - 1.89 (m, 1 H) 1.68 - 1.78 (m, 1 H) 1.52 - 1.61 (m, 1 H) 1.39 (br. s., 3 H)。 Step B: (3-((3- (4-Chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) Trihydroborate

N- (4-Chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.3 g, 1.018 mmol) in N, N-dimethylformamide (20 ml) was added to (3- ( Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.173 g, 1.018 mmol) was added. The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (60-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (3-((3- (4-chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2. ] Octane-1-yl) trihydroborate was obtained as a white powder (0.25 g, 0.63 mmol, 61.9% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 12.19 (s, 1 H) 9.72 (br. S., 1 H) 7.93 (d, J = 7.93 Hz, 1 H) 7.51 (d, J = 7.93 Hz, 1 H) 7.26-7.32 (m, 1 H) 5.32 (s, 1 H) 3.88 (dd, J = 13.73, 4.88 Hz, 1 H) 3.75 (dd, J = 13.73, 4.88 Hz, 1 H) 2.73 -2.95 (m, 6 H) 2.08 (br. S., 1 H) 1.96 (br. S., 1 H) 1.79-1.89 (m, 1 H) 1.68-1.78 (m, 1 H) 1.52-1.61 ( m, 1 H) 1.39 (br. s., 3 H).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−((３−(４−クロロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.２３ｇ, ０.５８mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１１７ml, ０.７５mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(２−(４−クロロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.１ｇ, ０.２７mmol, 収率４７.６％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.95 (s, 1 H) 7.79 (d, J=7.93 Hz, 1 H) 7.42 (d, J=7.63 Hz, 1 H) 7.18 (t, J=7.93 Hz, 1 H) 3.92 (d, J=10.38 Hz, 1 H) 3.77 (d, J=10.38 Hz, 1 H) 3.26 - 3.38 (m, 1 H) 3.19 (dd, J=15.26, 1.53 Hz, 1 H) 3.01 - 3.11 (m, 1 H) 2.81 - 3.00 (m, 3 H) 2.31 (br. s., 1 H) 2.03 - 2.15 (m, 1 H) 1.70 - 1.89 (m, 3 H) 1.45 (br. s., 3 H)。 Step C: (2- (4-Chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- Yl) trihydroborate

(3-((3- (4-Chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.] In N, N-dimethylformamide (20 ml). To 2] octane-1-yl) trihydroborate (0.23 g, 0.58 mmol) was added N, N′-diisopropylcarbodiimide (0.117 ml, 0.75 mmol). The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (2- (4-chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2]. ] Octane] -1'-yl) trihydroborate was obtained as a white powder (0.1 g, 0.27 mmol, 47.6% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.95 (s, 1 H) 7.79 (d, J = 7.93 Hz, 1 H) 7.42 (d, J = 7.63 Hz, 1 H) 7.18 (t, J = 7.93 Hz, 1 H) 3.92 (d, J = 10.38 Hz, 1 H) 3.77 (d, J = 10.38 Hz, 1 H) 3.26-3.38 (m, 1 H) 3.19 (dd, J = 15.26, 1.53 Hz , 1 H) 3.01-3.11 (m, 1 H) 2.81-3.00 (m, 3 H) 2.31 (br. S., 1 H) 2.03-2.15 (m, 1 H) 1.70-1.89 (m, 3 H) 1.45 (br. S., 3 H).

アセトン(９ml)中の(２−(４−クロロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.０８ｇ, ０.２２１mmol)に、３Ｍ ＨＣｌ(０.５５１ml, １.６５４mmol)を加えた。反応混合物を室温で４時間撹拌した。酢酸エチルを加え、水層を集め、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(４−クロロベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０５ｇ, ０.１４mmol, 収率６５.０％)。
¹H NMR(500 MHz, DMSO-d₆) δ ppm 8.90 (br. s., 1 H) 7.77 (dd, J=7.78, 1.07 Hz, 1 H) 7.41 (dd, J=7.78, 1.07 Hz, 1 H) 7.17 (t, J=7.78 Hz, 1 H) 3.94 (d, J=10.07 Hz, 1 H) 3.67 (d, J=10.07 Hz, 1 H) 3.00 - 3.11 (m, 2 H) 2.76 - 2.88 (m, 2 H) 2.68 (t, J=7.63 Hz, 2 H) 2.11 (br. s., 1 H) 1.91 - 2.00 (m, 1 H) 1.47 - 1.67 (m, 3 H). MS (LC/MS) R.T. = 2.11; [M+H]⁺ = 349.1。 Step D: N- (4-Chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (4-Chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane]-in acetone (9 ml) To 1′-yl) trihydroborate (0.08 g, 0.221 mmol) was added 3M 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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (4-chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine was obtained as a white powder (0.05 g, 0.14 mmol, yield 65.0%).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.90 (br. S., 1 H) 7.77 (dd, J = 7.78, 1.07 Hz, 1 H) 7.41 (dd, J = 7.78, 1.07 Hz, 1 H) 7.17 (t, J = 7.78 Hz, 1 H) 3.94 (d, J = 10.07 Hz, 1 H) 3.67 (d, J = 10.07 Hz, 1 H) 3.00-3.11 (m, 2 H) 2.76-2.88 (m, 2 H) 2.68 (t, J = 7.63 Hz, 2 H) 2.11 (br. s., 1 H) 1.91-2.00 (m, 1 H) 1.47-1.67 (m, 3 H). MS (LC / MS) RT = 2.11; [M + H] ⁺ = 349.1.

The enantiomers were separated using a Chiralcel OJ-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA). The wavelength was set at 220 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (4-chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-amine (0.11 g, 0.30 mmol, yield 34.8%).
(7a, S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.90 (br. S., 1 H) 7.77 (d, J = 7.32 Hz, 1 H) 7.41 (d, J = 7.93 Hz, 1 H) 7.17 (t, J = 7.93 Hz, 1 H) 3.94 (d, J = 10.07 Hz, 1 H) 3.67 (d, J = 10.07 Hz, 1 H) 3.00-3.11 (m, 2 H) 2.76-2.90 (m, 2 H) 2.68 (t, J = 7.78 Hz, 2 H) 2.11 (br. S., 1 H) 1.91-2.01 (m, 1 H) 1.47-1.68 (m, 3 H ). MS (LC / MS) RT = 2.06; [M + H] ⁺ = 349.1.

The second peak is (R) -N- (4-chlorobenzo [d] thiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane]- 2-Amine (0.11 g, 0.30 mmol, yield 35.2%).
(7b, R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.89 (br. S., 1 H) 7.77 (d, J = 7.93 Hz, 1 H) 7.40 (d, J = 7.93 Hz, 1 H) 7.16 (t, J = 7.93 Hz, 1 H) 3.94 (d, J = 10.07 Hz, 1 H) 3.67 (d, J = 10.07 Hz, 1 H) 2.99-3.11 (m, 2 H) 2.74-2.88 (m, 2 H) 2.68 (t, J = 7.78 Hz, 2 H) 2.11 (br. S., 1 H) 1.90-2.01 (m, 1 H) 1.44-1.68 (m, 3 H ). MS (LC / MS) RT = 2.07; [M + H] ⁺ = 349.1.

Example 8
N- (1H-benzo [d] imidazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の１Ｈ−ベンゾ[ｄ]イミダゾール−２−アミン(１.２８ｇ, ９.６１mmol)に、１,１'−チオカルボニルジイミダゾール(２.２２７ｇ, １２.５mmol)を加えた。反応混合物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で１時間乾燥させ、Ｎ−(１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た(１.８ｇ, ７.４mmol, 収率７７％)。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide

To 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 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 1 hour to give N- (1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide (1.8 g, 7 0.4 mmol, yield 77%). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、Ｎ−(１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(１.０７ｇ, ４.４mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.７４８ｇ, ４.４mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(６０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−((３−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルチオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(１.２８ｇ, ３.７１mmol, 収率８４％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.41 (s, 1 H), 11.16 (s, 1 H), 7.43 (d, J=3.05 Hz, 3 H), 7.13 (ddd, J=9.46, 3.81, 3.51 Hz, 3 H), 5.36 (s, 1 H), 4.01-4.07 (m, 3 H), 3.79 (dd, J=13.28, 4.12 Hz, 1 H), 2.83-2.92 (m, 6 H), 2.71 (d, J=14.04 Hz, 2 H), 2.04-2.13 (m, 2 H), 1.89-1.94 (m, 3 H), 1.74 (td, J=9.46, 5.49 Hz, 2 H), 1.51-1.59 (m, 2 H), 1.38 (s, 2 H), 1.31 (s, 1 H)。
ＬＣ／ＭＳにより、本ＬＣ／ＭＳ条件でＢＨ_３が無い生成物を確認した：保持時間 2.75 (M+1-BH₃ = 332.2)。 Step B: (3-((3-1H-benzo [d] imidazol-2-ylthioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) tri Hydroborate

N- (1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide (1.07 g, 4.4 mmol) in N, N-dimethylformamide (20 ml) was added to (3- (Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.748 g, 4.4 mmol) was added. The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (60-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (3-((3-1H-benzo [d] imidazol-2-ylthioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2]. Octan-1-yl) trihydroborate was obtained as a white powder (1.28 g, 3.71 mmol, 84% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.41 (s, 1 H), 11.16 (s, 1 H), 7.43 (d, J = 3.05 Hz, 3 H), 7.13 (ddd, J = 9.46 , 3.81, 3.51 Hz, 3 H), 5.36 (s, 1 H), 4.01-4.07 (m, 3 H), 3.79 (dd, J = 13.28, 4.12 Hz, 1 H), 2.83-2.92 (m, 6 H), 2.71 (d, J = 14.04 Hz, 2 H), 2.04-2.13 (m, 2 H), 1.89-1.94 (m, 3 H), 1.74 (td, J = 9.46, 5.49 Hz, 2 H) , 1.51-1.59 (m, 2 H), 1.38 (s, 2 H), 1.31 (s, 1 H).
LC / MS confirmed the product without BH _{3 under the} present LC / MS conditions: retention time 2.75 (M + 1-BH ₃ = 332.2).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−((３−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルチオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.０ｇ, ２.９mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.５８７ml, ３.７７mmol)を加えた。反応混合物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(２−(１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.８１ｇ, ２.６mmol, 収率９０％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 7.29 (s, 1 H), 6.99 - 7.05 (m, 2 H), 3.90 (t, J=9.77 Hz, 1 H), 3.75 (d, J=10.38 Hz, 1 H), 3.26 - 3.35 (m, 1 H), 3.13 (dd, J=14.95, 1.53 Hz, 1 H), 2.98 - 3.06 (m, 1 H), 2.84 - 2.92 (m, 3 H), 2.22 (s, 1 H), 1.98 - 2.05 (m, 1 H), 1.72 - 1.82 (m, 3 H), 1.45 (s, 1 H)。
ＬＣ／ＭＳにより、本ＬＣ／ＭＳ条件でＢＨ_３が無い生成物を確認した：保持時間 2.29 (M+1-BH₃ = 298.2)。 Step C: (2- (1H-Benzo [d] imidazol-2-ylamino) -4H-1'-ammoniospiro- [oxazole-5,3'-bicyclo [2.2.2] octane] -1 ' -Yl) trihydroborate

(3-((3-1H-benzo [d] imidazol-2-ylthioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.] In N, N-dimethylformamide (20 ml). To 2] octane-1-yl) trihydroborate (1.0 g, 2.9 mmol) was added N, N′-diisopropylcarbodiimide (0.587 ml, 3.77 mmol). The reaction mixture was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (2- (1H-benzo [d] imidazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2]. ] Octane] -1'-yl) trihydroborate was obtained as a white powder (0.81 g, 2.6 mmol, 90% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 7.29 (s, 1 H), 6.99-7.05 (m, 2 H), 3.90 (t, J = 9.77 Hz, 1 H), 3.75 (d, J = 10.38 Hz, 1 H), 3.26-3.35 (m, 1 H), 3.13 (dd, J = 14.95, 1.53 Hz, 1 H), 2.98-3.06 (m, 1 H), 2.84-2.92 (m, 3 H), 2.22 (s, 1 H), 1.98-2.05 (m, 1 H), 1.72-1.82 (m, 3 H), 1.45 (s, 1 H).
LC / MS confirmed the product without BH _{3 under these} LC / MS conditions: retention time 2.29 (M + 1-BH ₃ = 298.2).

アセトン(９ml)中の(２−(１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.７７ｇ, ２.５mmol)に、３Ｍ ＨＣｌ(６.２ml, １８.６mmol)を加えた。反応物を室温で４時間撹拌した。酢酸エチルを加え、水層を集めて、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせ、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.５ｇ, １.６８mmol, 収率６８％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.45 (s, 1 H), 9.20 (s, 1 H), 7.32 (s, 1 H), 7.00 (dd, J=5.65, 2.90 Hz, 4 H), 3.91 (d, J=10.07 Hz, 2 H), 3.64 (d, J=10.07 Hz, 2 H), 2.98 - 3.05 (m, 4 H), 2.73 - 2.82 (m, 4 H), 2.67 (t, J=7.63 Hz, 4 H), 2.03 (d, J=2.75 Hz, 2 H), 1.85 - 1.92 (m, 2 H), 1.54 - 1.63 (m, 4 H), 1.44 - 1.51 (m, 2 H). MS (LC/MS) R.T. = 1.30; [M+H]⁺ = 298.2。 Step D: N- (1H-Benzo [d] imidazol-2-yl) -4H-1′-azaspiro- [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (1H-Benzo [d] imidazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane]-in acetone (9 ml) To 1′-yl) trihydroborate (0.77 g, 2.5 mmol) was added 3M 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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (1H-benzo [d] imidazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine was obtained as a white powder (0.5 g, 1.68 mmol, 68% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.45 (s, 1 H), 9.20 (s, 1 H), 7.32 (s, 1 H), 7.00 (dd, J = 5.65, 2.90 Hz, 4 H), 3.91 (d, J = 10.07 Hz, 2 H), 3.64 (d, J = 10.07 Hz, 2 H), 2.98-3.05 (m, 4 H), 2.73-2.82 (m, 4 H), 2.67 (t, J = 7.63 Hz, 4 H), 2.03 (d, J = 2.75 Hz, 2 H), 1.85-1.92 (m, 2 H), 1.54-1.63 (m, 4 H), 1.44-1.51 (m MS (LC / MS) RT = 1.30; [M + H] ⁺ = 298.2.

The enantiomers were separated using a Chiralpak AS-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 330 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.11 g, 0.36 mmol, 33.0% were obtained.
(8a; R-isomer): mp 255 ° C. (decomposition). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.43 (br. S., 1 H) 9.16 (br. S., 1 H) 7.14-7.53 (m, 2 H) 6.82-7.09 (m, 2 H) 3.91 (d, J = 9.77 Hz, 1 H) 3.64 (d, J = 10.07 Hz, 1 H) 2.96-3.09 (m, 2 H ) 2.71-2.87 (m, 2 H) 2.62-2.73 (m, 2 H) 2.01-2.08 (m, 1 H) 1.81-1.97 (m, 1 H) 1.54-1.66 (m, 2 H) 1.40-1.53 ( m, 1 H). MS (LC / MS) RT = 1.26; [M + H] ⁺ = 298.2.

From the second peak, 0.11 g, 0.36 mmol, 33.0% were obtained.
(8b; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.43 (br. S., 1 H) 9.20 (br. S., 1 H) 7.10-7.60 (m, 2 H) 6.83-7.11 (m, 2 H) 3.91 (d, J = 9.77 Hz, 1 H) 3.64 (d, J = 9.77 Hz, 1 H) 2.94-3.16 (m, 2 H) 2.71-2.86 (m, 2 H) 2.59-2.72 (m, 2 H) 1.97-2.09 (m, 1 H) 1.81-1.95 (m, 1 H) 1.53-1.71 (m, 2 H) 1.41-1.53 (m, 1 H). MS (LC / MS) RT = 1.28; [M + H] ⁺ = 298.2.

Example 9
N- (6-Chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の６−クロロベンゾ[ｄ]チアゾール−２−アミン(１.１４ｇ, ６.１７mmol)に、１,１'−チオカルボニルジイミダゾール(１.４３ｇ, ８mmol)を加えた。反応物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で１時間乾燥させ、Ｎ−(６−クロロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(１.１６ｇ, ３.９mmol, 収率６４％)。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (6-Chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

To 1-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 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 1 hour, and N- (6-chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (1.16 g, 3.9 mmol, Yield 64%). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(６−クロロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.８６ｇ, ２.９mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５ｇ, ２.９mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(６０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−((３−(６−クロロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.４ｇ, １.０１mmol, 収率３４.６％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.92 (br. s., 1 H) 9.89 (br. s., 1 H) 8.08 (br. s., 1 H) 7.62 (br. s., 1 H) 7.40 - 7.50 (m, 1 H) 5.40 (br. s., 1 H) 3.88 (d, J=10.20 Hz, 1 H) 3.76 (d, J=10.20 Hz, 1 H) 2.67 - 3.02 (m, 6 H) 2.08 (br. s., 1 H) 1.80 - 1.95 (m, 2 H) 1.73 (br. s., 1 H) 1.01 - 1.63 (m, 4 H). MS (LC/MS) R.T. = 3.87; [M+H]⁺ = 395.1。 Step B: (3-((3- (6-Chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) Trihydroborate

N- (6-Chlorobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.86 g, 2.9 mmol) in N, N-dimethylformamide (20 ml) was added to (3- ( Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.5 g, 2.9 mmol) was added. The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (60-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo and (3-((3- (6-chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2]. ] Octan-1-yl) trihydroborate was obtained as a white powder (0.4 g, 1.01 mmol, 34.6% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.92 (br. S., 1 H) 9.89 (br. S., 1 H) 8.08 (br. S., 1 H) 7.62 (br. S. , 1 H) 7.40-7.50 (m, 1 H) 5.40 (br. S., 1 H) 3.88 (d, J = 10.20 Hz, 1 H) 3.76 (d, J = 10.20 Hz, 1 H) 2.67-3.02 (m, 6 H) 2.08 (br. s., 1 H) 1.80-1.95 (m, 2 H) 1.73 (br. s., 1 H) 1.01-1.63 (m, 4 H). MS (LC / MS ) RT = 3.87; [M + H] ⁺ = 395.1.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−((３−(６−クロロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.３７ｇ, ０.９３mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１９ml, １.２mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(２−(６−クロロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.１２ｇ, ０.３３mmol, 収率３５.５％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.10 (br. s., 1 H) 7.95 (d, J=2.14 Hz, 1 H) 7.60 (d, J=8.55 Hz, 1 H) 7.36 (dd, J=8.55, 2.14 Hz, 1 H) 3.88 (d, J=10.38 Hz, 1 H) 3.76 (d, J=10.38 Hz, 1 H) 3.26 - 3.37 (m, 1 H) 3.17 (dd, J=14.95, 1.83 Hz, 1 H) 3.00 - 3.11 (m, 1 H) 2.80 - 2.97 (m, 3 H) 2.28 (br. s., 1 H) 2.00 - 2.11 (m, 1 H) 1.69 - 1.88 (m, 3 H) 1.46 (br. s., 3 H). LC/MS: 保持時間 2.94 (M+1-BH₃ = 349.1)。 Step C: (2- (6-Chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- Yl) trihydroborate

(3-((3- (6-Chlorobenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.] In N, N-dimethylformamide (20 ml). To 2] octane-1-yl) trihydroborate (0.37 g, 0.93 mmol) was added N, N′-diisopropylcarbodiimide (0.19 ml, 1.2 mmol). The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (2- (6-chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2]. ] Octane] -1'-yl) trihydroborate was obtained as a white powder (0.12 g, 0.33 mmol, 35.5% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.10 (br.s., 1 H) 7.95 (d, J = 2.14 Hz, 1 H) 7.60 (d, J = 8.55 Hz, 1 H) 7.36 ( dd, J = 8.55, 2.14 Hz, 1 H) 3.88 (d, J = 10.38 Hz, 1 H) 3.76 (d, J = 10.38 Hz, 1 H) 3.26-3.37 (m, 1 H) 3.17 (dd, J = 14.95, 1.83 Hz, 1 H) 3.00-3.11 (m, 1 H) 2.80-2.97 (m, 3 H) 2.28 (br. S., 1 H) 2.00-2.11 (m, 1 H) 1.69-1.88 ( m, 3 H) 1.46 (br. s., 3 H). LC / MS: Retention time 2.94 (M + 1-BH ₃ = 349.1).

アセトン(９ml)中の、(２−(６−クロロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.１ｇ, ０.２８mmol)に、３Ｍ ＨＣｌ(０.６９ml, ２.０７mmol)を加えた。反応物を室温で４時間撹拌した。酢酸エチルを加え、水層を集めて、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機層を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(６−クロロベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０６ｇ, ０.１７mmol, 収率６２.４％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.03 (br. s., 1 H) 7.93 (d, J=2.14 Hz, 1 H) 7.58 (d, J=8.55 Hz, 1 H) 7.35 (dd, J=8.55, 2.14 Hz, 1 H) 3.90 (d, J=10.07 Hz, 1 H) 3.65 (d, J=10.07 Hz, 1 H) 2.99 - 3.11 (m, 2 H) 2.73 - 2.89 (m, 2 H) 2.67 (t, J=7.63 Hz, 2 H) 2.08 (br. s., 1 H) 1.82 - 1.99 (m, 1 H) 1.43 - 1.67 (m, 3 H). MS (LC/MS) R.T. = 2.07; [M+H]⁺ = 349.1。 Step D: N- (6-Chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (6-Chlorobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] in acetone (9 ml) To 1′-yl) trihydroborate (0.1 g, 0.28 mmol) was added 3M 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 × 40 ml). The organic layers were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (6-chlorobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′- Bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.06 g, 0.17 mmol, 62.4% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.03 (br.s., 1 H) 7.93 (d, J = 2.14 Hz, 1 H) 7.58 (d, J = 8.55 Hz, 1 H) 7.35 ( dd, J = 8.55, 2.14 Hz, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 2.99-3.11 (m, 2 H) 2.73-2.89 (m , 2 H) 2.67 (t, J = 7.63 Hz, 2 H) 2.08 (br. S., 1 H) 1.82-1.99 (m, 1 H) 1.43-1.67 (m, 3 H). MS (LC / MS ) RT = 2.07; [M + H] ⁺ = 349.1.

The enantiomers were separated using a Chiralpak AS-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 220 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.034 g, 0.10 mmol, 36.2% was obtained.
(9a; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.02 (br. S., 1 H) 7.93 (d, J = 2.14 Hz, 1 H) 7.58 (d, J = 8.55 Hz, 1 H) 7.35 (dd, J = 8.55, 2.14 Hz, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 2.97-3.11 (m , 2 H) 2.73-2.89 (m, 2 H) 2.67 (t, J = 7.48 Hz, 2 H) 2.08 (br. S., 1 H) 1.93 (br. S., 1 H) 1.43-1.69 (m , 3 H). MS (LC / MS) RT = 2.05; [M + H] ⁺ = 349.1. Optical rotation = + 4.00 °.

From the second peak, 0.037 g, 0.10 mmol, 39.4% were obtained.
(9b; R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.02 (br. S., 1 H) 7.93 (d, J = 2.14 Hz, 1 H) 7.58 (d, J = 8.55 Hz, 1 H) 7.35 (dd, J = 8.55, 2.44 Hz, 1 H) 3.90 (d, J = 10.07 Hz, 1 H) 3.65 (d, J = 10.07 Hz, 1 H) 2.93-3.13 (m , 2 H) 2.72-2.91 (m, 2 H) 2.62-2.73 (m, 2 H) 2.08 (br. S., 1 H) 1.93 (d, J = 1.22 Hz, 1 H) 1.44-1.68 (m, 3 H). MS (LC / MS) RT = 2.04; [M + H] ⁺ = 349.1. Optical rotation = -3.74 °.

Example 10
N- (1-methyl-1H-benzo [d] imidazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の、１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−アミン(１.２８ｇ, ８.７mmol)に、１,１'−チオカルボニルジイミダゾール(２.０１５ｇ, １１.３１mmol)を加えた。反応物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で１時間乾燥させ、Ｎ−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た(１.６ｇ, ６.２２mmol, 収率７１.５％)。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (1-Methyl-1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide

1-Methyl-1H-benzo [d] imidazol-2-amine (1.28 g, 8.7 mmol) in acetonitrile (30 ml) was added to 1,1′-thiocarbonyldiimidazole (2.015 g, 11.31 mmol). ) Was added. The reaction was stirred at 50 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 1 hour to give N- (1-methyl-1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide (1 .6 g, 6.22 mmol, yield 71.5%). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、Ｎ−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(１.０４ｇ, ４.０４mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.６８７ｇ, ４.０４mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(６０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−ヒドロキシ−３−((３−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(１.２８ｇ, ３.５６mmol, 収率８８％)。ＬＣ／ＭＳにより、本ＬＣ／ＭＳ条件でＢＨ_３が無い生成物を確認した：保持時間 3.01 (M+1-BH₃ = 346.2)。 Step B: (3-Hydroxy-3-((3- (1-methyl-1H-benzo [d] imidazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane- 1-yl) trihydroborate

To N- (1-methyl-1H-benzo [d] imidazol-2-yl) -1H-imidazole-1-carbothioamide (1.04 g, 4.04 mmol) in N, N-dimethylformamide (20 ml). (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.687 g, 4.04 mmol) was added. The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (60-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (3-hydroxy-3-((3- (1-methyl-1H-benzo [d] imidazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2 2.2.2] Octan-1-yl) trihydroborate was obtained as a white powder (1.28 g, 3.56 mmol, 88% yield). LC / MS confirmed the product without BH _{3 under the} present LC / MS conditions: retention time 3.01 (M + 1-BH ₃ = 346.2).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−ヒドロキシ−３−((３−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.１９ｇ, ３.３１mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(１.５５ml, ９.９mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(２−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.９４ｇ, ２.９mmol, 収率８７％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.36 (br. s., 1 H) 7.34 - 7.44 (m, 1 H) 7.26 - 7.34 (m, 1 H) 7.00 - 7.12 (m, 2 H) 3.90 (d, J=10.32 Hz, 1 H) 3.77 (d, J=10.32 Hz, 1 H) 3.57 (s, 3 H) 3.28 (dd, J=14.86, 2.27 Hz, 1 H) 3.13 (dd, J=14.86, 1.51 Hz, 1 H) 2.95 - 3.08 (m, 1 H) 2.75 - 2.95 (m, 3 H) 2.22 (br. s., 1 H) 1.96 - 2.11 (m, 1 H) 1.67 - 1.89 (m, 3 H) 1.43 (br. s., 3 H). LC/MS : 保持時間 2.37 (M+1-BH₃ = 312.2)。 Step C: (2- (1-Methyl-1H-benzo [d] imidazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1'-yl) trihydroborate

(3-Hydroxy-3-((3- (1-methyl-1H-benzo [d] imidazol-2-yl) thioureido) methyl) -1-ammoniobicyclo in N, N-dimethylformamide (20 ml) To [2.2.2] octan-1-yl) trihydroborate (1.19 g, 3.31 mmol) was added N, N′-diisopropylcarbodiimide (1.55 ml, 9.9 mmol). The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (2- (1-methyl-1H-benzo [d] imidazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2 2.2.2] Octane] -1′-yl) trihydroborate was obtained as a white powder (0.94 g, 2.9 mmol, 87% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.36 (br. S., 1 H) 7.34-7.44 (m, 1 H) 7.26-7.34 (m, 1 H) 7.00-7.12 (m, 2 H ) 3.90 (d, J = 10.32 Hz, 1 H) 3.77 (d, J = 10.32 Hz, 1 H) 3.57 (s, 3 H) 3.28 (dd, J = 14.86, 2.27 Hz, 1 H) 3.13 (dd, J = 14.86, 1.51 Hz, 1 H) 2.95-3.08 (m, 1 H) 2.75-2.95 (m, 3 H) 2.22 (br. S., 1 H) 1.96-2.11 (m, 1 H) 1.67-1.89 (m, 3 H) 1.43 (br. s., 3 H). LC / MS: retention time 2.37 (M + 1-BH ₃ = 312.2).

アセトン(９ml)中の、(２−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.９２ｇ, ２.８mmol)に、３Ｍ ＨＣｌ(７.１ml, ２１.２mmol)を加えた。反応物を室温で４時間撹拌した。酢酸エチルを加え、水層を集めて、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(１−メチル−１Ｈ−ベンゾ[ｄ]イミダゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.８４ｇ, ２.７mmol, 収率９５％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.24 - 9.39 (m, 1 H) 7.33 - 7.42 (m, 1 H) 7.25 - 7.33 (m, 1 H) 6.99 - 7.10 (m, 2 H) 3.92 (d, J=10.07 Hz, 1 H) 3.66 (d, J=10.07 Hz, 1 H) 3.57 (s, 3 H) 2.97 - 3.08 (m, 2 H) 2.78 (t, J=7.81 Hz, 2 H) 2.67 (t, J=7.81 Hz, 2 H) 2.01 - 2.09 (m, 1 H) 1.80 - 1.96 (m, 1 H) 1.40 - 1.66 (m, 3 H). MS (LC/MS) R.T. = 1.49; [M+H]⁺ = 312.2。 Step D: N- (1-Methyl-1H-benzo [d] imidazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(2- (1-Methyl-1H-benzo [d] imidazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] in acetone (9 ml). To 2] octane] -1′-yl) trihydroborate (0.92 g, 2.8 mmol) was added 3M 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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (1-methyl-1H-benzo [d] imidazol-2-yl) -4H-1′-azaspiro [oxazole-5, 3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.84 g, 2.7 mmol, 95% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.24-9.39 (m, 1 H) 7.33-7.42 (m, 1 H) 7.25-7.33 (m, 1 H) 6.99-7.10 (m, 2 H) 3.92 (d, J = 10.07 Hz, 1 H) 3.66 (d, J = 10.07 Hz, 1 H) 3.57 (s, 3 H) 2.97-3.08 (m, 2 H) 2.78 (t, J = 7.81 Hz, 2 H) 2.67 (t, J = 7.81 Hz, 2 H) 2.01-2.09 (m, 1 H) 1.80-1.96 (m, 1 H) 1.40-1.66 (m, 3 H). MS (LC / MS) RT = 1.49; [M + H] ⁺ = 312.2.

The enantiomers were separated using a Chiralcel OJ-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 22% methanol in CO ₂ (0.1% DEA) while monitoring UV at 300 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.065 g, 0.205 mmol, 39.8% were obtained.
(10a, R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.31 (br. S., 1 H) 7.35-7.45 (m, 1 H) 7.26-7.34 (m, 1 H ) 7.00-7.15 (m, 2 H) 3.93 (d, J = 10.10 Hz, 1 H) 3.67 (d, J = 10.10 Hz, 1 H) 3.59 (s, 3 H) 2.93-3.13 (m, 2 H) 2.74-2.89 (m, 2 H) 2.61-2.74 (m, 2 H) 2.05 (br. S., 1 H) 1.91 (br. S., 1 H) 1.38-1.68 (m, 3 H). MS ( LC / MS) RT = 1.37; [M + H] ⁺ = 312.2. Optical rotation =-16.02 °.

From the second peak, 0.06 g, 0.19 mmol, 36.8% were obtained.
(10b; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.31 (br. S., 1 H) 7.33-7.43 (m, 1 H) 7.25-7.33 (m, 1 H ) 6.93-7.11 (m, 2 H) 3.93 (dd, J = 9.92, 3.20 Hz, 1 H) 3.67 (dd, J = 9.92, 3.20 Hz, 1 H) 3.58 (s, 3 H) 2.94-3.13 (m , 2 H) 2.74-2.85 (m, 2 H) 2.59-2.72 (m, 2 H) 2.04 (br. S., 1 H) 1.90 (br. S., 1 H) 1.37-1.70 (m, 3 H ). MS (LC / MS) RT = 1.37; [M + H] ⁺ = 312.2. Optical rotation = + 35.99 °.

Example 11
N- (6-Ethoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

バイアルに、アセトニトリル(１５ml)中の６−エトキシベンゾ[ｄ]チアゾール−２−アミン(１.５ｇ, ７.７２mmol)およびジ(１Ｈ−イミダゾール−１−イル)メタンチオン(１.７８９ｇ, １０.０４mmol)を入れた。反応物を８０℃で一夜加熱した。反応混合物を濾過し、沈殿物を集めて、２.４ｇ(７.８８mmol, １０２％)の赤錆色の固体を得た。 Step A: N- (6-Ethoxybenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

A vial is charged with 6-ethoxybenzo [d] thiazol-2-amine (1.5 g, 7.72 mmol) and di (1H-imidazol-1-yl) methanethione (1.789 g, 10.04 mmol) in acetonitrile (15 ml). ) The reaction was heated at 80 ° C. overnight. The reaction mixture was filtered and the precipitate was collected to give 2.4 g (7.88 mmol, 102%) of a red rust solid.

バイアルに、Ｎ,Ｎ−ジメチルホルムアミド(８ml)中の、Ｎ−(６−エトキシベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(２.４ｇ, ７.８８mmol)および(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.３４１ｇ, ７.８８mmol)を入れた。反応物を８０℃まで加熱した。２時間後、反応物を水およびクロロホルムに注ぎ、有機物を抽出し、濃縮し、赤色の油状物を得た。この物質を、さらに精製せず、または特性決定せずに、次の反応に用いた。 Step B: (3-((3- (6-Ethoxybenzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl Trihydroborate

In a vial, N- (6-ethoxybenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (2.4 g, 7.88 mmol) and N, N-dimethylformamide (8 ml) and (3- (Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (1.341 g, 7.88 mmol) was added. The reaction was heated to 80 ° C. After 2 hours, the reaction was poured into water and chloroform, the organics were extracted and concentrated to give a red oil. This material was used in the next reaction without further purification or characterization.

フラスコに、Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中の、(３−((３−(６−エトキシベンゾ[ｄ]チアゾール−２−イル)チオウレイド)−メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(３.２ｇ, ７.９mmol)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(４.３ml, ２７.６mmol)に入れた。反応物を７０℃で２時間加熱し、水およびクロロホルムに注いだ。有機物を集め、濃縮して残渣を得た。残渣をエーテル中で磨砕し、吸引濾過して沈殿物を集め、１.１１ｇ(２.９８mmol, ３７.８％)の灰色の粉末を得た。 Step C: (2- (6-Ethoxybenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro- [oxazole-5,3′-bicyclo [2.2.2] octane] -1 '-Yl) trihydroborate

The flask was charged with (3-((3- (6-ethoxybenzo [d] thiazol-2-yl) thioureido) -methyl) -3-hydroxy-1-ammonio in N, N-dimethylformamide (10 ml). Bicyclo [2.2.2] octan-1-yl) trihydroborate (3.2 g, 7.9 mmol) and N, N′-diisopropylcarbodiimide (4.3 ml, 27.6 mmol) were added. The reaction was heated at 70 ° C. for 2 hours and poured into water and chloroform. The organics were collected and concentrated to give a residue. The residue was triturated in ether and suction filtered to collect the precipitate, yielding 1.11 g (2.98 mmol, 37.8%) of a gray powder.

バイアルに、アセトン(１０ml)中の、(２−(６−エトキシベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(５００mg, １.３４mmol)およびＨＣｌ(８.０６ml, ２４.１７mmol)を入れた。反応をＨＰＬＣによってモニターした。２時間後、ＬＣ／ＭＳによって反応が完了したことが示された。反応物を水およびクロロホルムに注ぎ、有機層を除去した。水層を中和し、クロロホルム(２×)で抽出した。二番目のクロロホルムフラクションを濃縮して白色の残渣を得た。固体をエーテル中で磨砕し、沈殿物を集め、３１４.４mg(０.８７７mmol, ６５.３％)の望ましい物質を得た。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.89 (1 H, br. s.), 7.50 (1 H, d, J=8.81 Hz), 7.38 (1 H, d, J=2.52 Hz), 6.91 (1 H, dd, J=8.81, 2.52 Hz), 4.03 (2 H, q, J=7.05 Hz), 3.87 (1 H, d, J=10.07 Hz), 3.62 (1 H, d, J=10.07 Hz), 3.02 (2 H, s), 2.78 (2 H, t, J=7.81 Hz), 2.66 (2 H, t, J=7.68 Hz), 2.05 (1 H, br. s.), 1.91 (1 H, br. s.), 1.42 - 1.67 (3 H, m), 1.33 (3 H, t, J=7.05 Hz). MS (LC/MS) R.T. = 1.60; [M+H]⁺ = 359.0。 Step D: N- (6-Ethoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

A vial is filled with (2- (6-ethoxybenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2. 2] Octane] -1'-yl) trihydroborate (500 mg, 1.34 mmol) and HCl (8.06 ml, 24.17 mmol) were added. The reaction was monitored by HPLC. After 2 hours, LC / MS indicated that the reaction was complete. The reaction was poured into water and chloroform and the organic layer was removed. The aqueous layer was neutralized and extracted with chloroform (2x). The second chloroform fraction was concentrated to give a white residue. The solid was triturated in ether and the precipitate was collected to give 314.4 mg (0.877 mmol, 65.3%) of the desired material.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.89 (1 H, br. S.), 7.50 (1 H, d, J = 8.81 Hz), 7.38 (1 H, d, J = 2.52 Hz) , 6.91 (1 H, dd, J = 8.81, 2.52 Hz), 4.03 (2 H, q, J = 7.05 Hz), 3.87 (1 H, d, J = 10.07 Hz), 3.62 (1 H, d, J = 10.07 Hz), 3.02 (2 H, s), 2.78 (2 H, t, J = 7.81 Hz), 2.66 (2 H, t, J = 7.68 Hz), 2.05 (1 H, br.s.), 1.91 (1 H, br. S.), 1.42-1.67 (3 H, m), 1.33 (3 H, t, J = 7.05 Hz). MS (LC / MS) RT = 1.60; [M + H] ⁺ = 359.0.

The enantiomers were separated using a Chiralpak AS-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 300 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 62.4 mg, 0.17 mmol, 31.2% were obtained.
(11a; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.87 (1 H, br. S.), 7.49 (1 H, d, J = 8.55 Hz), 7.38 (1 H, d, J = 2.44 Hz), 6.91 (1 H, dd, J = 8.85, 2.44 Hz), 4.03 (2 H, q, J = 7.02 Hz), 3.87 (1 H, d, J = 10.07 Hz) , 3.61 (1 H, d, J = 10.07 Hz), 3.02 (2 H, s), 2.72-2.85 (2 H, m), 2.66 (2 H, t, J = 7.63 Hz), 2.05 (1 H, br. s.), 1.91 (1 H, br. s.), 1.43-1.64 (3 H, m), 1.33 (3 H, t, J = 7.02 Hz). MS (LC / MS) RT = 1.81; [M + H] ⁺ = 359.1.

From the second peak, 58.9 mg, 0.164 mmol, 29.5% were obtained.
(11b; R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.86 (1 H, br. S.), 7.48 (1 H, d, J = 8.85 Hz), 7.37 (1 H, d, J = 2.44 Hz), 6.91 (1 H, d, J = 2.75 Hz), 4.02 (2 H, q, J = 7.02 Hz), 3.86 (1 H, d, J = 9.77 Hz), 3.61 (1 H, d, J = 9.77 Hz), 3.01 (2 H, s), 2.72-2.85 (2 H, m), 2.62-2.69 (2 H, m), 2.04 (1 H, d, J = 2.44 Hz), 1.90 (1 H, d, J = 4.27 Hz), 1.42-1.63 (3 H, m), 1.29-1.35 (3 H, m). MS (LC / MS) RT = 1.52; (M + H ] ⁺ = 359.1.

Example 12
N- (6-Methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の６−メチルベンゾ[ｄ]チアゾール−２−アミン(１.ｇ, ６.２mmol)に、１,１'−チオカルボニルジイミダゾール(１.４４ｇ, ８.１mmol)を加えた。反応物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で１時間乾燥させ、Ｎ−(６−メチルベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た(１.０６ｇ, ３.８６mmol, 収率６２％)。これをさらに精製せず、または特性決定せずに、次の工程に用いた。 Step A: N- (6-Methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

To 1-methylbenzo [d] thiazol-2-amine (1.g, 6.2mmol) in acetonitrile (30ml) was added 1,1'-thiocarbonyldiimidazole (1.44g, 8.1mmol). The reaction was stirred at 50 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° C.) for 1 hour to give N- (6-methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (1.06 g, 3 .86 mmol, 62% yield). This was used in the next step without further purification or characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、Ｎ−(６−メチルベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.９６ｇ, ３.５mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５９５ｇ, ３.５mmol)を加えた。反応物を７０℃で４時間撹拌した。反応混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(４０〜１００％ 酢酸エチル／ヘキサン)。生成物のフラクションを真空で濃縮し、(３−ヒドロキシ−３−((３−(６−メチルベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.８８ｇ, ２.３３８mmol, 収率６６.８％)。
MS (LC/MS) R.T. = 3.71; [M+H]⁺ = 375.2。 Step B: (3-Hydroxy-3-((3- (6-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) Trihydroborate

N- (6-Methylbenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.96 g, 3.5 mmol) in N, N-dimethylformamide (20 ml) was added to (3- (Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.595 g, 3.5 mmol) was added. The reaction was stirred at 70 ° C. for 4 hours. The reaction mixture was concentrated and purified by silica gel chromatography (40-100% ethyl acetate / hexanes). The product fractions were concentrated in vacuo to give (3-hydroxy-3-((3- (6-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2. ] Octan-1-yl) trihydroborate was obtained as a white powder (0.88 g, 2.338 mmol, 66.8% yield).
MS (LC / MS) RT = 3.71; [M + H] ⁺ = 375.2.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−ヒドロキシ−３−((３−(６−メチルベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.８６ｇ, ２.２８５mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.２８８ｇ, ２.２８５mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、酢酸エチルを加えた。沈殿物を濾過し、さらに酢酸エチルで洗浄した。粉末を真空オーブン(７０℃)中で乾燥させ、(２−(６−メチルベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.６５ｇ, １.８９９mmol, 収率８３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.03 (1 H, br. s.), 7.59 (1 H, s), 7.51 (1 H, d, J=8.24 Hz), 7.15 (1 H, d, J=8.24 Hz), 3.86 (1 H, d, J=10.38 Hz), 3.74 (1 H, d, J=10.38 Hz), 3.29 (1 H, dd, J=15.26, 1.83 Hz), 3.14 (1 H, d, J=15.26 Hz), 2.99 - 3.09 (1 H, m), 2.80 - 2.95 (3 H, m), 2.36 (3 H, s), 2.25 (1 H, br. s.), 2.03 (1 H, t, J=10.22 Hz), 1.70 - 1.85 (3 H, m). MS (LC/MS) R.T. = 2.78; [M+H]⁺ = 343.2。 Step C: (2- (6-Methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- Yl) trihydroborate

(3-Hydroxy-3-((3- (6-methylbenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.] In N, N-dimethylformamide (20 ml). To 2] octane-1-yl) trihydroborate (0.86 g, 2.285 mmol) was added N, N′-diisopropylcarbodiimide (0.288 g, 2.285 mmol). The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated and ethyl acetate was added. The precipitate was filtered and further washed with ethyl acetate. The powder was dried in a vacuum oven (70 ° C.) and (2- (6-methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2. 2.2] octane] -1′-yl) trihydroborate was obtained as a white powder (0.65 g, 1.899 mmol, 83% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.03 (1 H, br.s.), 7.59 (1 H, s), 7.51 (1 H, d, J = 8.24 Hz), 7.15 (1 H , d, J = 8.24 Hz), 3.86 (1 H, d, J = 10.38 Hz), 3.74 (1 H, d, J = 10.38 Hz), 3.29 (1 H, dd, J = 15.26, 1.83 Hz), 3.14 (1 H, d, J = 15.26 Hz), 2.99-3.09 (1 H, m), 2.80-2.95 (3 H, m), 2.36 (3 H, s), 2.25 (1 H, br. S. ), 2.03 (1 H, t, J = 10.22 Hz), 1.70-1.85 (3 H, m). MS (LC / MS) RT = 2.78; [M + H] ⁺ = 343.2.

アセトン(９ml)中の(２−(６−メチルベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.２４ｇ, ０.７０mmol)に、３Ｍ ＨＣｌ(１.７５３ml, ５.２６mmol)を加えた。反応混合物を室温で４時間撹拌した。酢酸エチルを加え、水層を集めて、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、Ｎ−(６−メチルベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.１９ｇ, ０.５８mmol, 収率８３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.96 (1 H, br. s.), 7.57 (1 H, s), 7.49 (1 H, d, J=8.24 Hz), 7.13 (1 H, d, J=8.24 Hz), 3.88 (1 H, d, J=10.07 Hz), 3.63 (1 H, d, J=9.77 Hz), 2.98 - 3.04 (2 H, m), 2.72 - 2.85 (2 H, m), 2.66 (2 H, t, J=7.63 Hz), 2.36 (3 H, s), 2.05 (1 H, d, J=2.14 Hz), 1.91 (1 H, br. s.), 1.53 - 1.64 (2 H, m), 1.42 - 1.53 (1 H, m). MS (LC/MS) R.T. = 1.79; [M+H]⁺ = 329.2。 Step D: N- (6-Methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (6-Methylbenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane]-in acetone (9 ml) To 1′-yl) trihydroborate (0.24 g, 0.70 mmol) was added 3M 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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and N- (6-methylbenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.19 g, 0.58 mmol, 83% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.96 (1 H, br. S.), 7.57 (1 H, s), 7.49 (1 H, d, J = 8.24 Hz), 7.13 (1 H , d, J = 8.24 Hz), 3.88 (1 H, d, J = 10.07 Hz), 3.63 (1 H, d, J = 9.77 Hz), 2.98-3.04 (2 H, m), 2.72-2.85 (2 H, m), 2.66 (2 H, t, J = 7.63 Hz), 2.36 (3 H, s), 2.05 (1 H, d, J = 2.14 Hz), 1.91 (1 H, br.s.), 1.53-1.64 (2 H, m), 1.42-1.53 (1 H, m). MS (LC / MS) RT = 1.79; [M + H] ⁺ = 329.2.

Enantiomers were separated using a Chiralcel OJ-H (4.6 × 25 cm, 5 μm) column with 30% methanol in CO ₂ (0.1% DEA) while monitoring UV at 300 nm. The separated peak was concentrated in vacuo to give a white powder. From the first peak of the column, 0.11 g, 0.34 mmol, 55% were obtained.
(12a; R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.95 (s, 1 H), 7.57 (s, 2 H), 7.48 (d, J = 8.24 Hz, 2 H ), 7.13 (d, J = 8.24 Hz, 2 H), 3.89 (d, J = 10.38 Hz, 2 H), 3.63 (d, J = 10.38 Hz, 2 H), 2.99-3.06 (m, 4 H) , 2.75-2.84 (m, 4 H), 2.67 (t, J = 7.63 Hz, 4 H), 2.37 (s, 7 H), 2.06 (s, 2 H), 1.92 (s, 2 H), 1.55- 1.64 (m, 4 H), 1.49 (dd, J = 9.77, 2.44 Hz, 2 H). MS (LC / MS) RT = 1.80; [M + H] ⁺ = 329.2. Optical rotation = -4.52.

From the second peak, 0.11 g, 0.34 mmol, 55% were obtained.
(12b; S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.96 (s, 1 H), 7.58 (s, 1 H), 7.49 (d, J = 8.24 Hz, 1 H ), 7.14 (d, J = 8.24 Hz, 1 H), 3.89 (d, J = 10.07 Hz, 1 H), 3.63 (d, J = 10.07 Hz, 1 H), 3.03 (d, J = 2.44 Hz, 2 H), 2.75-2.84 (m, 2 H), 2.67 (t, J = 7.78 Hz, 2 H), 2.37 (s, 3 H), 2.06 (s, 1 H), 1.92 (s, 1 H) , 1.55-1.64 (m, 2 H), 1.49 (dd, J = 9.77, 2.75 Hz, 1 H). MS (LC / MS) RT = 1.80; [M + H] ⁺ = 329.2. Optical rotation = + 10.08 .

Example 13
2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazol-6-ol

アセトニトリル(３０ml)中の、６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−アミン(国際公開第2007/086800号102頁に記載された通りに製造)(３.１ｇ, １１.０５mmol)に、チオカルボニル ジイミダゾール(２.５６ｇ, １４.３７mmol)を加えた。反応物を７０℃まで一夜加熱した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリルで洗浄し、黄色の固体を得た。生成物であるＮ−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(３.８５ｇ, ９.８６mmol, 収率８９％)をさらに精製することなく次の工程に用いた。
MS (LC/MS) R.T. = 2.56; [M+H]⁺ = 388.9。 Step A: N- (6- (tert-butyldimethylsilyloxy) benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

6- (tert-Butyldimethylsilyloxy) benzo [d] thiazol-2-amine (prepared as described in WO 2007/086800, page 102) in acetonitrile (30 ml) (3.1 g, 11 To .05 mmol) was added thiocarbonyldiimidazole (2.56 g, 14.37 mmol). The reaction was heated to 70 ° C. overnight. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile to give a yellow solid. The product N- (6- (tert-butyldimethylsilyloxy) benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (3.85 g, 9.86 mmol, 89% yield) Was used in the next step without further purification.
MS (LC / MS) RT = 2.56; [M + H] ⁺ = 388.9.

Ｎ,Ｎ−ジメチルホルムアミド(４０ml)中のＮ−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(３.８５ｇ, ９.８６mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.６８ｇ, ９.８６mmol)を加えた。反応物を８０℃で２時間加熱した。反応物を冷却し、クロロホルムおよび水の混合物に注いだ。有機層を集め、真空で濃縮し、(３−((３−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを黄色の油状物として得た(６.２ｇ)。 Step B: (3-((3- (6- (tert-Butyldimethylsilyloxy) benzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoniobicyclo [2.2. 2] Octane-1-yl) trihydroborate

N- (6- (tert-Butyldimethylsilyloxy) benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (3.85 g, 9. 9. g) in N, N-dimethylformamide (40 ml). 86 mmol) was added (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (1.68 g, 9.86 mmol). The reaction was heated at 80 ° C. for 2 hours. The reaction was cooled and poured into a mixture of chloroform and water. The organic layer was collected and concentrated in vacuo to give (3-((3- (6- (tert-butyldimethylsilyloxy) benzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1-ammoni. Obicyclo [2.2.2] octan-1-yl) trihydroborate was obtained as a yellow oil (6.2 g).

Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中の、(３−((３−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(４.９ｇ, ９.９mmol)に、１,３−ジイソプロピルカルボジイミド(５.４ml, ３４.５mmol)を加えた。反応物を８０℃に加熱し、ＬＣ／ＭＳによってモニターした。反応物を冷却し、クロロホルムおよび水の混合物に注いだ。有機層を集めて、真空で濃縮した。残った残渣をエーテル中で磨砕した。沈殿物を集めて、(２−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを得た(３.２８ｇ, ７.１５mmol, 収率７２.６％)。
MS (LC/MS) R.T. = 3.60; [M+H-BH₃]⁺ = 445.2。 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

(3-((3- (6- (tert-Butyldimethylsilyloxy) benzo [d] thiazol-2-yl) thioureido) methyl) -3-hydroxy-1 in N, N-dimethylformamide (10 ml) To ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (4.9 g, 9.9 mmol) was added 1,3-diisopropylcarbodiimide (5.4 ml, 34.5 mmol). The reaction was heated to 80 ° C. and monitored by LC / MS. The reaction was cooled and 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 and (2- (6- (tert-butyldimethylsilyloxy) benzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2 2.2.2] Octane] -1′-yl) trihydroborate was obtained (3.28 g, 7.15 mmol, yield 72.6%).
MS (LC / MS) RT = 3.60; [M + H-BH 3] + = 445.2.

アセトン(１０ml)中の、(２−(６−(ｔｅｒｔ−ブチルジメチルシリルオキシ)ベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(３.２ｇ, ６.９８mmol)に、ＨＣｌ(８.１４ml, ２４.４３mmol)を加えた。反応物を室温で３時間撹拌した。混合物を水に注ぎ、飽和重炭酸ナトリウムで中和した。水層をクロロホルムで抽出した。有機層を集め、真空で濃縮した。残った残渣をエーテル中で磨砕し、ラセミの２−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)ベンゾ[ｄ]チアゾール−６−オールを白色の粉末として得た(９５８mg, ２.９０mmol, 収率４１.５％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.36 (1 H, br. s.), 8.85 (1 H, br. s.), 7.41 (1 H, d, J=8.85 Hz), 7.12 (1 H, d, J=2.44 Hz), 6.78 (1 H, dd, J=8.85, 2.44 Hz), 3.86 (1 H, d, J=10.07 Hz), 3.61 (1 H, d, J=10.07 Hz), 2.96 - 3.06 (2 H, m), 2.71 - 2.85 (2 H, m), 2.66 (2 H, t, J=7.78 Hz), 2.04 (1 H, br. s.), 1.90 (1 H, br. s.), 1.43 - 1.64 (3 H, m). MS (LC/MS) R.T. = 1.03; [M+H]⁺ = 331.29。 Step D: 2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazol-6-ol

(2- (6- (tert-Butyldimethylsilyloxy) benzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [] in acetone (10 ml). To 2.2.2] octane] -1′-yl) trihydroborate (3.2 g, 6.98 mmol) 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 extracted with chloroform. The organic layer was collected and concentrated in vacuo. The remaining residue was triturated in ether to give racemic 2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazole. -6-ol was obtained as a white powder (958 mg, 2.90 mmol, 41.5% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.36 (1 H, br. S.), 8.85 (1 H, br. S.), 7.41 (1 H, d, J = 8.85 Hz), 7.12 (1 H, d, J = 2.44 Hz), 6.78 (1 H, dd, J = 8.85, 2.44 Hz), 3.86 (1 H, d, J = 10.07 Hz), 3.61 (1 H, d, J = 10.07 Hz), 2.96-3.06 (2 H, m), 2.71-2.85 (2 H, m), 2.66 (2 H, t, J = 7.78 Hz), 2.04 (1 H, br.s.), 1.90 (1 H, br. S.), 1.43-1.64 (3 H, m). MS (LC / MS) RT = 1.03; [M + H] ⁺ = 331.29.

The enantiomers were separated using a Chiralpak AD-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA). The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazole-6 -Ol (395.2 mg, 1.19 mmol, 41.4% yield).
(13a, S-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.36 (1 H, br. S.), 8.84 (1 H, br. S.), 7.41 (1 H, d, J = 8.55 Hz), 7.12 (1 H, d, J = 2.44 Hz), 6.78 (1 H, dd, J = 8.55, 2.44 Hz), 3.86 (1 H, d, J = 9.77 Hz), 3.61 (1 H, d, J = 10.07 Hz), 2.96-3.05 (2 H, m), 2.72-2.84 (2 H, m), 2.65 (2 H, t, J = 7.63 Hz), 2.04 (1 H, br. s.), 1.90 (1 H, br. s.), 1.52-1.64 (2 H, m), 1.43-1.52 (1 H, m) .MS (LC / MS) RT = 1.30; (M + H] ⁺ = 331.4.

The second peak is (R) -2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazole-6- All (375 mg, 1.14 mmol, 39.3% yield).
(13b, R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 7.40 (1 H, d, J = 8.55 Hz), 7.11 (1 H, d, J = 2.44 Hz), 6.77 (1 H, dd, J = 8.55, 2.44 Hz), 3.86 (1 H, d, J = 10.07 Hz), 3.60 (1 H, d, J = 10.07 Hz), 3.01 (2 H, s), 2.73- 2.84 (2 H, m), 2.65 (2 H, t, J = 7.78 Hz), 2.04 (1 H, br. S.), 1.90 (1 H, br. S.), 1.54-1.62 (2 H, m), 1.43-1.52 (1 H, m). MS (LC / MS) RT = 1.43; [M + H] ⁺ = 331.4.

Example 14
N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

アセトニトリル(２０ml)中の４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−アミン(０.５ｇ, ３.２４mmol)に、ジ(１Ｈ−イミダゾール−１−イル)メタンチオン(０.５８ｇ, ３.２４mmol)を加えた。反応物を５０℃で４時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、Ｎ−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを白色の粉末として得た(０.６０ｇ, ２.２７mmol, 収率７０.０％)。 Step A: N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

4,5,6,7-Tetrahydrobenzo [d] thiazol-2-amine (0.5 g, 3.24 mmol) in acetonitrile (20 ml) was added to di (1H-imidazol-1-yl) methanethione (0.58 g). , 3.24 mmol) was added. The reaction was stirred at 50 ° C. for 4 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexane) and N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -1H-imidazole-1- Carbothioamide was obtained as a white powder (0.60 g, 2.27 mmol, 70.0% yield).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.５４ｇ, ２mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.３５ｇ, ２mmol)を加えた。反応物を５０℃で３時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、最初のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて、濃縮し、(３−ヒドロキシ−３−((３−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.５１ｇ, １.３９mmol, 収率６８.２％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 11.46 (s, 1 H), 5.26 (s, 1 H), 3.89 (dd, J=13.58, 5.34 Hz, 1 H), 3.68 (dd, J=13.73, 4.88 Hz, 1 H), 2.55 - 2.94 (m, 8 H), 2.06 (dd, J=9.31, 3.20 Hz, 1 H), 1.66 - 1.90 (m, 6 H), 1.21 - 1.59 (m, 4 H); [M+H]⁺ = 365.1。 Step B: (3-Hydroxy-3-((3- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2 ] Octane-1-yl) trihydroborate

To N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.54 g, 2 mmol) in N, N-dimethylformamide (20 ml) (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.35 g, 2 mmol) was added. The reaction was stirred at 50 ° C. for 3 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexanes) to give the first spot / fraction (TLC) as product. Fractions were combined and concentrated to give (3-hydroxy-3-((3- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [ 2.2.2] octan-1-yl) trihydroborate was obtained as a white powder (0.51 g, 1.39 mmol, 68.2% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 11.46 (s, 1 H), 5.26 (s, 1 H), 3.89 (dd, J = 13.58, 5.34 Hz, 1 H), 3.68 (dd, J = 13.73, 4.88 Hz, 1 H), 2.55-2.94 (m, 8 H), 2.06 (dd, J = 9.31, 3.20 Hz, 1 H), 1.66-1.90 (m, 6 H), 1.21-1.59 (m, 4 H); [M + H] ⁺ = 365.1.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(３−ヒドロキシ−３−((３−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５ｇ, １.３７mmol)に、Ｎ,Ｎ−ジイソプロピルカルボジイミド(０.７４ml, ４.７８mmol)を加えた。反応物を７０℃で４時間撹拌した。反応物を濃縮し、粗製の残渣を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、二番目のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて濃縮し、(２−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.３８ｇ, １.１４mmol, 収率８４％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.26 - 8.86 (m, 1 H), 3.78 (d, J=9.46 Hz, 1 H), 3.66 (d, J=9.77 Hz, 1 H), 3.19 - 3.30 (m, J=14.95, 2.14 Hz, 1 H), 2.96 - 3.12 (m, 2 H), 2.78 - 2.94 (m, 3 H), 2.54 - 2.65 (m, 4 H), 2.19 (s, 1 H), 2.00 (s, 1 H), 1.66 - 1.85 (m, 7 H), 1.43 (m, 3 H). MS (LC/MS) R.T. = 2.50; [M+H-BH₃]⁺ = 319.1。 Step C: (2- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -1'-yl) trihydroborate

(3-Hydroxy-3-((3- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) thioureido) methyl) -1-ammoni in N, N-dimethylformamide (20 ml) To obicyclo [2.2.2] octan-1-yl) trihydroborate (0.5 g, 1.37 mmol) was added N, N-diisopropylcarbodiimide (0.74 ml, 4.78 mmol). The reaction was stirred at 70 ° C. for 4 hours. The reaction was concentrated to give a crude residue. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexane) to give a second spot / fraction (TLC) as product. The fractions were combined and concentrated to give (2- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2]. 2.2.2] Octane] -1′-yl) trihydroborate was obtained as a white powder (0.38 g, 1.14 mmol, 84% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.26-8.86 (m, 1 H), 3.78 (d, J = 9.46 Hz, 1 H), 3.66 (d, J = 9.77 Hz, 1 H), 3.19 -3.30 (m, J = 14.95, 2.14 Hz, 1 H), 2.96-3.12 (m, 2 H), 2.78-2.94 (m, 3 H), 2.54-2.65 (m, 4 H), 2.19 (s, 1 H), 2.00 (s, 1 H), 1.66-1.85 (m, 7 H), 1.43 (m, 3 H). MS (LC / MS) RT = 2.50; [M + H-BH ₃ ] ⁺ = 319.1.

アセトン(９ml)中の(２−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.３６ｇ, １.０８mmol)に、３Ｍ ＨＣｌ(０.３６ml, １.０８mmol)を加えた。反応物を室温で４時間撹拌した。ＴＬＣによって反応の完了が示された(より低いスポット)。酢酸エチルを加え、水層を分離した。水層を１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(４,５,６,７−テトラヒドロベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.２８ｇ, ０.８４mmol, 収率７８％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.30 - 8.75 (br.s, 1 H), 3.79 (d, J=9.44 Hz , 1 H), 3.55 (d, J=9.76 Hz , 1 H), 2.90 - 3.03 (m, 2 H), 2.54 - 2.84 (m, 8 H), 2.00 (s, 1 H), 1.81 - 1.93 (m, 1 H), 1.75 (d, J=2.44 Hz, 4 H), 1.36 - 1.67 (m, 3 H). MS (LC/MS) R.T. = 1.39; [M+H]⁺ = 319.1。 Step D: N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (4,5,6,7-Tetrahydrobenzo [d] thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2. To 2.2] octane] -1′-yl) trihydroborate (0.36 g, 1.08 mmol) was added 3M HCl (0.36 ml, 1.08 mmol). The reaction was stirred at room temperature for 4 hours. TLC showed completion of reaction (lower spot). Ethyl acetate was added and the aqueous layer was separated. The aqueous layer was neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and racemic N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -4H-1′-azaspiro. [Oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.28 g, 0.84 mmol, 78% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.30-8.75 (br.s, 1 H), 3.79 (d, J = 9.44 Hz, 1 H), 3.55 (d, J = 9.76 Hz, 1 H) , 2.90-3.03 (m, 2 H), 2.54-2.84 (m, 8 H), 2.00 (s, 1 H), 1.81-1.93 (m, 1 H), 1.75 (d, J = 2.44 Hz, 4 H ), 1.36-1.67 (m, 3 H). MS (LC / MS) RT = 1.39; [M + H] ⁺ = 319.1.

The enantiomers were separated using a Chiralpak AD-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 23% methanol in CO ₂ (0.1% DEA). The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] octane] -2-amine (0.1 g, 0.29 mmol, 36.8% yield).
(14a, S-isomer): ¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.29-8.89 (br.s, 1 H), 3.80 (d, J = 9.46 Hz, 1 H), 3.54 (d , J = 9.77 Hz, 1 H), 2.94-3.04 (m, 2 H), 2.55-2.85 (m, 8 H), 2.00 (s, 1 H), 1.85-1.94 (m, 1 H), 1.70- 1.81 (m, J = 2.43 Hz, 4 H), 1.39-1.65 (m, 3 H). MS (LC / MS) RT = 1.54; [M + H] ⁺ = 319.1.

The second peak is (R) -N- (4,5,6,7-tetrahydrobenzo [d] thiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2 2.2.2 Octane] -2-amine (0.11 g, 0.30 mmol, 38.2% yield).
(14b, R-isomer): ¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.24-8.96 (br.s, 1 H), 3.79 (d, J = 9.46 Hz, 1 H), 3.52 (d , J = 9.76 Hz, 1 H), 2.95-3.08 (m, 2 H), 2.55-2.80 (m, 8 H), 2.00 (s, 1 H), 1.85-1.90 (m, 1 H), 1.70- 1.79 (m, J = 2.42 Hz, 4 H), 1.39-1.63 (m, 3 H); [M + H] ⁺ = 319.1.

Example 15
N- (4-Isopropylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)中の４−イソプロピルチアゾール−２−アミン(１.０４ｇ, ７.３１mmol)に、１,１−チオカルボニルジイミダゾール(１.７ｇ, ９.５mmol)を加えた。反応物を５０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、アセトニトリル(２×５０ml)で洗浄した。黄色の粉末を真空オーブン(４０℃)中で２時間乾燥させた。生成物であるＮ−(４−イソプロピルチアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(１.０２ｇ, ４.０４mmol, 収率５５.３％)を、さらに精製することなく次の工程に直接用いた。 Step A: N- (4-Isopropylthiazol-2-yl) -1H-imidazole-1-carbothioamide

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 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with acetonitrile (2 × 50 ml). The yellow powder was dried in a vacuum oven (40 ° 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 obtained without further purification. Used directly in the process.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、Ｎ−(４−イソプロピルチアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.５７ｇ, ２.２６mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.３８ｇ, ２.２６mmol)を加えた。反応物を７０℃で２４時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、(３−ヒドロキシ−３−((３−(４−イソプロピルチアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.５６ｇ, １.５８mmol, 収率７０.０％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.62 (s, 1 H), 6.70 (s, 1 H), 5.31 (s, 1 H), 3.58 - 3.96 (m, 2 H), 2.63 - 3.11 (m, 7 H), 2.00 - 2.22 (m, 1 H), 1.65 - 1.97 (m, 3 H), 1.16 - 1.61 (m, 10 H). MS (LC/MS) R.T. = 3.43; [M+H]⁺ = 353.2。 Step B: (3-Hydroxy-3-((3- (4-isopropylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate

N- (4-Isopropylthiazol-2-yl) -1H-imidazole-1-carbothioamide (0.57 g, 2.26 mmol) in N, N-dimethylformamide (20 ml) was added to (3- (aminomethyl ) -3-Hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.38 g, 2.26 mmol) was added. The reaction was stirred at 70 ° C. for 24 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexane), (3-hydroxy-3-((3- (4-isopropylthiazol-2-yl) thioureido) methyl) -1-ammoni. Obicyclo [2.2.2] octan-1-yl) trihydroborate was obtained as a white powder (0.56 g, 1.58 mmol, 70.0% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.62 (s, 1 H), 6.70 (s, 1 H), 5.31 (s, 1 H), 3.58-3.96 (m, 2 H), 2.63- 3.11 (m, 7 H), 2.00-2.22 (m, 1 H), 1.65-1.97 (m, 3 H), 1.16-1.61 (m, 10 H). MS (LC / MS) RT = 3.43; (M + H] ⁺ = 353.2.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−ヒドロキシ−３−((３−(５−イソプロピルチアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５４ｇ, １.５２mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.８３ml, ５.３３mmol)を加えた。反応物を５０℃で２４時間撹拌した。反応物を濃縮し、粗製の残渣を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、二番目のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて、濃縮し、(２−(５−イソプロピルチアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.３９ｇ, １.２２mmol, 収率８０％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 6.55 (s, 1 H), 3.75 (m, 2 H), 3.20 - 3.31 (m, 1 H), 2.96 - 3.15 (m, 2 H), 2.77 - 2.97 (m, 4 H), 2.22 (s, 1 H), 2.01 (s, 1 H), 1.68 - 1.90 (m, 3 H), 1.43 (s, 3 H), 1.21 (d, J=7.02 Hz, 6 H). MS (LC/MS) R.T. = 2.36; [M+H-BH₃]⁺ = 307.2。 Step C: (2- (5-Isopropylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) tri Hydroborate

(3-Hydroxy-3-((3- (5-isopropylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] in N, N-dimethylformamide (20 ml). To octan-1-yl) trihydroborate (0.54 g, 1.52 mmol) was added N, N′-diisopropylcarbodiimide (0.83 ml, 5.33 mmol). The reaction was stirred at 50 ° C. for 24 hours. The reaction was concentrated to give a crude residue. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexane) to give a second spot / fraction (TLC) as product. Fractions were combined and concentrated to give (2- (5-isopropylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1 '-Yl) trihydroborate was obtained as a white powder (0.39 g, 1.22 mmol, 80% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 6.55 (s, 1 H), 3.75 (m, 2 H), 3.20-3.31 (m, 1 H), 2.96-3.15 (m, 2 H), 2.77-2.97 (m, 4 H), 2.22 (s, 1 H), 2.01 (s, 1 H), 1.68-1.90 (m, 3 H), 1.43 (s, 3 H), 1.21 (d, J = 7.02 Hz, 6 H). MS (LC / MS) RT = 2.36; [M + H-BH ₃ ] ⁺ = 307.2.

アセトン(９ml)中の(２−(４−イソプロピルチアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.４２ｇ, １.３１mmol)に、３Ｍ ＨＣｌ(０.４４ml, １.３１mmol)を加えた。反応物を室温で４時間撹拌した。ＴＬＣにより、反応の完了が示された(より低いスポット)。酢酸エチルを加え、水層を分離した。水層を１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(４−イソプロピルチアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.３４ｇ, １.０５mmol, 収率８０％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.20 - 8.91 (m, 1 H), 6.52 (s, 1 H), 3.83 (d, J=9.46 Hz, 1 H), 3.57 (d, J=9.46 Hz, 1 H), 2.99 (s, 2 H), 2.58 - 2.92 (m, 5 H), 2.02 (s, 1 H), 1.82 - 1.96 (m, 1 H), 1.38 - 1.66 (m, 3 H), 1.20 (d, J=7.02 Hz, 6 H). MS (LC/MS) R.T. = 1.27; [M+H]⁺ = 307.1。 Step D: N- (4-Isopropylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (4-Isopropylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1′- in acetone (9 ml) Yl) trihydroborate (0.42 g, 1.31 mmol) was added 3M HCl (0.44 ml, 1.31 mmol). The reaction was stirred at room temperature for 4 hours. TLC showed completion of reaction (lower spot). Ethyl acetate was added and the aqueous layer was separated. The aqueous layer was neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and racemic N- (4-isopropylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine was obtained as a white powder (0.34 g, 1.05 mmol, 80% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.20-8.91 (m, 1 H), 6.52 (s, 1 H), 3.83 (d, J = 9.46 Hz, 1 H), 3.57 (d, J = 9.46 Hz, 1 H), 2.99 (s, 2 H), 2.58-2.92 (m, 5 H), 2.02 (s, 1 H), 1.82-1.96 (m, 1 H), 1.38-1.66 (m, 3 H), 1.20 (d, J = 7.02 Hz, 6 H). MS (LC / MS) RT = 1.27; [M + H] ⁺ = 307.1.

The enantiomers were separated using a Chiralcel OJ-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 23% methanol in CO ₂ (0.1% DEA). The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (4-isopropylthiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine (0.01 g, 0.03 mmol, 2.95% yield).
(15a, S-isomer): ¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.35-8.95 (m, 1 H), 6.52 (s, 1 H), 3.82 (d, J = 9.41 Hz, 1 H), 3.57 (d, J = 9.46 Hz, 1 H), 2.99 (s, 2 H), 2.60-2.88 (m, 5 H), 2.02 (s, 2 H), 1.82-1.96 (m, 1 H ), 1.38-1.65 (m, 3 H), 1.20 (d, J = 6.71 Hz, 6 H). MS (LC / MS) RT = 1.38; [M + H] ⁺ = 307.1.

The second peak is (R) -N- (4-isopropylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- It was an amine (0.02 g, 0.05 mmol, 4.83% yield).
(15b, R-isomer): ¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.35-8.91 (m, 1 H), 6.52 (s, 1 H), 3.82 (d, J = 9.44 Hz, 1 H), 3.57 (d, J = 9.46 Hz, 1 H), 2.99 (s, 2 H), 2.60-2.90 (m, 5 H), 2.02 (s, 2 H), 1.82-1.96 (m, 1 H ), 1.38-1.62 (m, 3 H), 1.20 (d, J = 6.78 Hz, 6 H). MS (LC / MS) RT = 1.49; [M + H] ⁺ = 307.3.

Example 16
N- (thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)およびテトラヒドロフラン(５ml)中の、チアゾール−２−アミン(２.１２ｇ, ２１.１７mmol)に、ジ(１Ｈ−イミダゾール−１−イル)メタンチオン(４.９０ｇ, ２７.５mmol)を加えた。反応物を６０℃で５時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、冷アセトニトリル(２×１５ml)で洗浄し、橙色〜褐色の粉末を得た。生成物であるＮ−(チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド (３.７０ｇ, １７.６０mmol, 収率８３％)を、さらに特性決定することなく次の工程に直接用いた。 Step A: N- (thiazol-2-yl) -1H-imidazole-1-carbothioamide

Di (1H-imidazol-1-yl) methanethione (4.90 g, 27.5 mmol) was added to thiazol-2-amine (2.12 g, 21.17 mmol) in acetonitrile (30 ml) and tetrahydrofuran (5 ml). It was. The reaction was stirred at 60 ° C. for 5 hours. The reaction was cooled to room temperature and the precipitate was filtered and washed with cold acetonitrile (2 × 15 ml) to give an orange-brown powder. The product N- (thiazol-2-yl) -1H-imidazole-1-carbothioamide (3.70 g, 17.60 mmol, 83% yield) was used directly in the next step without further characterization. It was.

Ｎ,Ｎ−ジメチルホルムアミド(３０ml)中の、Ｎ−(チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(１.７ｇ, ８mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.３７ｇ, ８mmol)を加えた。反応物を５０℃で４時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、最初のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて、濃縮し、(３−ヒドロキシ−３−((３−チアゾール−２−イルチオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(１.５ｇ, ４.８０mmol, 収率５９.８％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.64 (s, 1 H), 7.42 (d, J=3.36 Hz, 1 H), 7.14 (m, 1 H), 5.32 (s, 1 H), 3.78 (dd, 2 H), 2.59 - 3.02 (m, 6 H), 1.99 - 2.18 (m, 1 H), 1.79 - 1.92 (m, 2 H), 1.64 - 1.80 (m, 1 H), 1.19 - 1.65 (m, 4 H). MS (LC/MS) R.T. = 2.73; [M+H]⁺ = 311.1。 Step B: (3-Hydroxy-3-((3-thiazol-2-ylthioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate

N- (thiazol-2-yl) -1H-imidazole-1-carbothioamide (1.7 g, 8 mmol) in N, N-dimethylformamide (30 ml) was added to (3- (aminomethyl) -3-hydroxy. -1-Ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (1.37 g, 8 mmol) was added. The reaction was stirred at 50 ° C. for 4 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexanes) to give the first spot / fraction (TLC) as product. The fractions were combined and concentrated to give (3-hydroxy-3-((3-thiazol-2-ylthioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydro. The borate was obtained as a white powder (1.5 g, 4.80 mmol, 59.8% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.64 (s, 1 H), 7.42 (d, J = 3.36 Hz, 1 H), 7.14 (m, 1 H), 5.32 (s, 1 H) , 3.78 (dd, 2 H), 2.59-3.02 (m, 6 H), 1.99-2.18 (m, 1 H), 1.79-1.92 (m, 2 H), 1.64-1.80 (m, 1 H), 1.19 -1.65 (m, 4 H). MS (LC / MS) RT = 2.73; [M + H] ⁺ = 311.1.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−ヒドロキシ−３−((３−チアゾール−２−イルチオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１.２ｇ, ３.８４mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(２.０９ml, １３.４５mmol)を加えた。反応物を５０℃で２４時間撹拌した。反応物を濃縮し、粗製の残渣を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、最初のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて、濃縮し、(２−(チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.８４ｇ, ３.０２mmol, 収率７９％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.37 - 9.14 (m, 1 H), 7.32 (d, J=3.66 Hz, 1 H), 7.04 (d, J=3.66 Hz, 2 H), 3.79 (d, J=10.07 Hz, 1 H), 3.67 (d, J=10.07 Hz, 1 H), 3.20 - 3.29 (m, J=14.95, 2.14 Hz, 1 H), 2.97 - 3.15 (m, 2 H), 2.78 - 2.94 (m, 3 H), 2.22 (s, 1 H), 1.95 - 2.08 (m, 1 H), 1.66 - 1.85 (m, 3 H), 1.43 (s, 3 H). MS (LC/MS) R.T. = 1.57; [M+H-BH₃]⁺ = 265.1。 Step C: (2- (Thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) trihydroborate

(3-Hydroxy-3-((3-thiazol-2-ylthioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1- in N, N-dimethylformamide (20 ml) Yl) trihydroborate (1.2 g, 3.84 mmol) was added N, N′-diisopropylcarbodiimide (2.09 ml, 13.45 mmol). The reaction was stirred at 50 ° C. for 24 hours. The reaction was concentrated to give a crude residue. The crude material was purified by flash chromatography (50-100% ethyl acetate-hexanes) to give the first spot / fraction (TLC) as product. The fractions were combined and concentrated to give (2- (thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl. ) Trihydroborate was obtained as a white powder (0.84 g, 3.02 mmol, 79% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.37-9.14 (m, 1 H), 7.32 (d, J = 3.66 Hz, 1 H), 7.04 (d, J = 3.66 Hz, 2 H), 3.79 (d, J = 10.07 Hz, 1 H), 3.67 (d, J = 10.07 Hz, 1 H), 3.20-3.29 (m, J = 14.95, 2.14 Hz, 1 H), 2.97-3.15 (m, 2 H ), 2.78-2.94 (m, 3 H), 2.22 (s, 1 H), 1.95-2.08 (m, 1 H), 1.66-1.85 (m, 3 H), 1.43 (s, 3 H). MS ( LC / MS) RT = 1.57; [M + H-BH 3] + = 265.1.

アセトン(９ml)中の(２−(チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.５７ｇ, ２.０５mmol)に、３Ｍ ＨＣｌ(０.６８ml, ２.０５mmol)を加えた。反応物を室温で４時間撹拌した。ＴＬＣにより、反応物の完了が示された(より低いスポット)。酢酸エチルを加え、水層を分離した。水層を１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.４ｇ, １.４４mmol, 収率７０.２％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.64 (s, 1 H), 7.24 (d, J=3.74 Hz, 2 H), 7.01 (d, J=3.75 Hz, 2 H), 3.78 (d, J=9.80 Hz, 2 H), 3.53 (d, J=9.80 Hz, 2 H), 2.97 - 3.05 (m, 4 H), 2.74 - 2.86 (m, 4 H), 2.65 (t, J=7.84 Hz, 4 H), 2.01 (s, 2 H), 1.88 (s, 2 H), 1.53 - 1.64 (m, 4 H), 1.45 - 1.56 (m, 2 H). MS (LC/MS) R.T. = 0.28; [M+H]⁺ = 265.1。 Step D: N- (thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (Thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) tri in acetone (9 ml) To hydroborate (0.57 g, 2.05 mmol) was added 3M HCl (0.68 ml, 2.05 mmol). The reaction was stirred at room temperature for 4 hours. TLC showed completion of the reaction (lower spot). Ethyl acetate was added and the aqueous layer was separated. The aqueous layer was neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 40 ml). The combined organics were dried over magnesium sulfate, filtered, concentrated in vacuo, and racemic N- (thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2. .2] Octane] -2-amine was obtained as a white powder (0.4 g, 1.44 mmol, yield 70.2%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.64 (s, 1 H), 7.24 (d, J = 3.74 Hz, 2 H), 7.01 (d, J = 3.75 Hz, 2 H), 3.78 ( d, J = 9.80 Hz, 2 H), 3.53 (d, J = 9.80 Hz, 2 H), 2.97-3.05 (m, 4 H), 2.74-2.86 (m, 4 H), 2.65 (t, J = 7.84 Hz, 4 H), 2.01 (s, 2 H), 1.88 (s, 2 H), 1.53-1.64 (m, 4 H), 1.45-1.56 (m, 2 H). MS (LC / MS) RT = 0.28; [M + H] ⁺ = 265.1.

The enantiomers were separated using a Chiralcel OJ-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 23% methanol in CO ₂ (0.1% DEA). The wavelength was set at 300 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine (0.12 g, 0.43 mmol, 18.80% yield).
(16a, S-isomer): ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.64 (s, 1 H), 7.26 (d, J = 3.75 Hz, 2 H), 7.00 (d, J = 3.77 Hz, 2 H), 3.78 (d, J = 9.81 Hz, 2 H), 3.54 (d, J = 9.82 Hz, 2 H), 2.95-3.10 (m, 4 H), 2.75-2.82 (m, 4 H), 2.65 (t, J = 7.80 Hz, 4 H), 2.01 (s, 2 H), 1.88 (s, 2 H), 1.53-1.60 (m, 4 H), 1.42-1.51 (m, 2 H ). MS (LC / MS) RT = 0.32; [M + H] ⁺ = 265.1.

The second peak was (R) -N- (thiazol-2-yl) -4H-1'-azaspiro [oxazol-5,3'-bicyclo [2.2.2] octane] -2-amine. (0.15 g, 0.52 mmol, yield 22.96%).
(16b, R-isomer): ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.64 (s, 1 H), 7.25 (d, J = 3.74 Hz, 2 H), 7.00 (d, J = 3.76 Hz, 2 H), 3.78 (d, J = 9.80 Hz, 2 H), 3.53 (d, J = 9.80 Hz, 2 H), 2.95-3.08 (m, 4 H), 2.75-2.84 (m, 4 H), 2.65 (t, J = 7.80 Hz, 4 H), 2.01 (s, 2 H), 1.88 (s, 2 H), 1.54-1.62 (m, 4 H), 1.43-1.53 (m, 2 H ). MS (LC / MS) RT = 0.28; [M + H] ⁺ = 265.3.

Example 17
N- (4- (4-Methoxyphenyl) -5-methylthiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(２５ml)中の４−(４−メトキシフェニル)−５−メチルチアゾール−２−アミン(０.９８ｇ, ４.４５mmol)に、ジ(１Ｈ−イミダゾール−１−イル)メタンチオン(１.０３ｇ, ５.７８mmol)を加えた。反応物を５０℃で３時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過した。粉末をアセトニトリル(２×１０ml)で洗浄し、乾燥させ、ラセミのＮ−(４−(４−メトキシフェニル)−５−メチルチアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを黄色の粉末として得た(１.２８ｇ, ３.８７mmol, 収率８７％)。生成物を次の工程に直接用いた。 Step A: N- (4- (4-Methoxyphenyl) -5-methylthiazol-2-yl) -1H-imidazole-1-carbothioamide

4- (4-Methoxyphenyl) -5-methylthiazol-2-amine (0.98 g, 4.45 mmol) in acetonitrile (25 ml) was added to di (1H-imidazol-1-yl) methanethione (1.03 g, 5.78 mmol) was added. The reaction was stirred at 50 ° C. for 3 hours. The reaction was cooled to room temperature and the precipitate was filtered. The powder was washed with acetonitrile (2 × 10 ml), dried and racemic N- (4- (4-methoxyphenyl) -5-methylthiazol-2-yl) -1H-imidazole-1-carbothioamide Obtained as a powder (1.28 g, 3.87 mmol, 87% yield). The product was used directly in the next step.

Ｎ,Ｎ−ジメチルホルムアミド(２５ml)中の、Ｎ−(４−(４−メトキシフェニル)−５−メチルチアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボキサミド(０.４４ｇ, １.３９mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.２４ｇ, １.３９mmol)を加えた。反応物を７０℃で２時間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.６５ml, ４.１６mmol)を加え、反応物を７５℃で２時間加熱した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をカラムクロマトグラフィーによって精製し(６０〜１００％ 酢酸エチル／ヘキサン)、(２−(４−(４−メトキシフェニル)−５−メチルチアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを黄色の粉末として得た(０.４１ｇ, １.０２９mmol, 収率７４.２％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 7.59 (d, J=7.63 Hz, 2 H), 6.98 (d, J=8.55 Hz, 2 H), 3.80 (s, 4 H), 3.67 (s, 1 H), 3.24 - 3.31 (m, 1 H), 3.13 (s, 1 H), 3.03 (s, 1 H), 2.83 - 2.92 (m, 3 H), 2.39 (s, 3 H), 2.23 (s, 1 H), 2.07 (s, 1 H), 1.73 - 1.82 (m, 2 H), 1.44 (s, 1 H). MS (LC/MS) R.T. = 2.88; [M+H]⁺ = 399.34。 Step B: (2- (4- (4-Methoxyphenyl) -5-methylthiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -1'-yl) trihydroborate

N- (4- (4-Methoxyphenyl) -5-methylthiazol-2-yl) -1H-imidazole-1-carboxamide (0.44 g, 1.39 mmol) in N, N-dimethylformamide (25 ml) To (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.24 g, 1.39 mmol) was added. The reaction was stirred at 70 ° C. for 2 hours. N, N′-diisopropylcarbodiimide (0.65 ml, 4.16 mmol) was added and the reaction was heated at 75 ° C. for 2 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by column chromatography (60-100% ethyl acetate / hexane), (2- (4- (4-methoxyphenyl) -5-methylthiazol-2-ylamino) -4H-1′-ammoni). Ospiro [oxazol-5,3′-bicyclo [2.2.2] octane] -1′-yl) trihydroborate was obtained as a yellow powder (0.41 g, 1.029 mmol, yield 74.2). %).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 7.59 (d, J = 7.63 Hz, 2 H), 6.98 (d, J = 8.55 Hz, 2 H), 3.80 (s, 4 H), 3.67 (s , 1 H), 3.24-3.31 (m, 1 H), 3.13 (s, 1 H), 3.03 (s, 1 H), 2.83-2.92 (m, 3 H), 2.39 (s, 3 H), 2.23 (s, 1 H), 2.07 (s, 1 H), 1.73-1.82 (m, 2 H), 1.44 (s, 1 H). MS (LC / MS) RT = 2.88; [M + H] ⁺ = 399.34.

アセトン(９ml)中の(３−ヒドロキシ−３−((３−(４−(４−メトキシフェニル)−５−メチルチアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.０８ｇ, ０.１９mmol)に、２Ｍ ＨＣｌ(０.０９ml, ０.１９mmol)を加えた。反応物を室温で１時間撹拌した。ＴＬＣにより、反応の完了が示された(より低いスポット)。酢酸エチルを加え、水層を集めて、１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(４−(４−メトキシフェニル)−５−メチルチアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０５ｇ, ０.１２mmol, 収率６６.８％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 7.58 (d, J=8.24 Hz, 2 H), 6.98 (d, J=8.55 Hz, 2 H), 3.83 (d, J=9.16 Hz, 1 H), 3.80 (s, 3 H), 3.56 (d, J=9.46 Hz, 1 H), 3.00 (s, 2 H), 2.74 - 2.83 (m, 2 H), 2.66 (t, J=7.63 Hz, 2 H), 2.38 (s, 3 H), 2.03 (s, 1 H), 1.91 (s, 1 H), 1.54 - 1.62 (m, 2 H), 1.48 (d, J=7.02 Hz, 1 H). MS (LC/MS) R.T. = 2.04; [M+H]⁺ = 385.28。 Step C: N- (4- (4-methoxyphenyl) -5-methylthiazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane]- 2-Amine

(3-Hydroxy-3-((3- (4- (4-methoxyphenyl) -5-methylthiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2 in acetone (9 ml). .2] octane-1-yl) trihydroborate (0.08 g, 0.19 mmol) was added 2M HCl (0.09 ml, 0.19 mmol). The reaction was stirred at room temperature for 1 hour. TLC showed completion of reaction (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 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and racemic N- (4- (4-methoxyphenyl) -5-methylthiazol-2-yl) -4H-1′-azaspiro [ Oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.05 g, 0.12 mmol, 66.8% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 7.58 (d, J = 8.24 Hz, 2 H), 6.98 (d, J = 8.55 Hz, 2 H), 3.83 (d, J = 9.16 Hz, 1 H ), 3.80 (s, 3 H), 3.56 (d, J = 9.46 Hz, 1 H), 3.00 (s, 2 H), 2.74-2.83 (m, 2 H), 2.66 (t, J = 7.63 Hz, 2 H), 2.38 (s, 3 H), 2.03 (s, 1 H), 1.91 (s, 1 H), 1.54-1.62 (m, 2 H), 1.48 (d, J = 7.02 Hz, 1 H) MS (LC / MS) RT = 2.04; [M + H] ⁺ = 385.28.

Example 18
(E) -N- (1′-azaspiro [oxazolidine-5,3′-bicyclo [2.2.2] octane] -2-ylidene) pyridin-3-amine

テトラヒドロフラン(３ml)中の(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(２４７mg, １.４５mmol)の懸濁液に、撹拌しながら、テトラヒドロフラン(１.５ml)中の３−イソチオシアナトピリジン(２９８mg, ２.１９mmol)の溶液を加え、反応混合物を室温で一夜撹拌した。反応混合物を真空で蒸発させ、残渣(白色の蝋状固体)をカラムクロマトグラフィーによって精製し(３％ メタノール／酢酸エチル)、(３−ヒドロキシ−３−((３−ピリジン−３−イルチオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の泡状物質として得た(２９４.６mg, ０.９６mmol, 収率６６.２％)。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 8.56 - 8.70 (m, 1 H), 8.24 - 8.36 (m, 1 H), 8.08 - 8.16 (m, 1 H), 7.32 - 7.48 (m, 1 H), 4.02 - 4.18 (m, 1 H), 3.67 - 3.77 (m, 1 H), 3.53 - 3.62 (m, 1 H), 2.88 - 3.11 (m, 3 H), 2.70 - 2.88 (m, 1 H), 2.15 - 2.29 (m, 1 H), 1.92 - 2.12 (m, 2 H), 1.73 - 1.89 (m, 1 H), 1.54 - 1.69 (m, 2 H). MS (LC/MS) R.T. = 1.00; [MH⁺-BH₃] = 293.10。 Step A: (3-Hydroxy-3-((3-pyridin-3-ylthioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate

Suspension of (3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate (247 mg, 1.45 mmol) in tetrahydrofuran (3 ml) To a stirred solution of 3-isothiocyanatopyridine (298 mg, 2.19 mmol) in tetrahydrofuran (1.5 ml) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated in vacuo and the residue (white waxy solid) was purified by column chromatography (3% methanol / ethyl acetate), (3-hydroxy-3-((3-pyridin-3-ylthioureido). ) Methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate as a white foam (294.6 mg, 0.96 mmol, 66.2% yield) .
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 8.56-8.70 (m, 1 H), 8.24-8.36 (m, 1 H), 8.08-8.16 (m, 1 H), 7.32-7.48 (m, 1 H), 4.02-4.18 (m, 1 H), 3.67-3.77 (m, 1 H), 3.53-3.62 (m, 1 H), 2.88-3.11 (m, 3 H), 2.70-2.88 (m, 1 H), 2.15-2.29 (m, 1 H), 1.92-2.12 (m, 2 H), 1.73-1.89 (m, 1 H), 1.54-1.69 (m, 2 H). MS (LC / MS) ^{RT = 1.00; [MH + -BH} 3] = 293.10.

Ｎ,Ｎ−ジメチルホルムアミド(５ml)中の(３−ヒドロキシ−３−((３−ピリジン−３−イルチオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(２９４.６mg, ０.９６mmol)の溶液に、Ｎ,Ｎ−ジメチルホルムアミド(１ml)中のＮ,Ｎ'−メタンジイリデンジプロパン−２−アミン(１２１mg, ０.９６mmol)の溶液を加え、反応混合物を室温で７日間静置した。さらに、０.５mlのＮ,Ｎ−ジメチルホルムアミド中の１３３mgのＮ,Ｎ'−メタンジイリデンジプロパン−２−アミンを加え、さらに７日間反応を続けた。反応物をカラムクロマトグラフィーによって精製し(５〜１０％ メタノール／酢酸エチル)、(Ｅ)−(２−(ピリジン−３−イルイミノ)−１'−アンモニオスピロ[オキサゾリジン−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを得た(１３４.３mg, ０.４９mmol, 収率５１.３％)。
¹H NMR (500 MHz, MeOD) δ ppm 8.34 - 8.60 (m, 1 H), 8.16 (d, J=4.27 Hz, 1 H), 7.59 - 7.96 (m, 1 H), 7.35 (dd, J=8.24, 4.88 Hz, 1 H), 3.89 (br. s., 1 H), 3.65 (d, J=9.46 Hz, 1 H), 3.16 - 3.23 (m, 1 H), 3.04 - 3.16 (m, 1 H), 2.85 - 3.04 (m, 2 H), 2.23 (br. s., 1 H), 1.74 - 1.97 (m, 4 H), 1.36 - 1.70 (m, 2 H). MS (LC/MS) R.T. = 0.65; [M+H-BH₃]⁺ = 259.21。 Step B: (E)-(2- (Pyridin-3-ylimino) -1′-ammoniospiro [oxazolidine-5,3′-bicyclo [2.2.2] octane] -1′-yl) trihydro Borate

(3-Hydroxy-3-((3-pyridin-3-ylthioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1-yl in N, N-dimethylformamide (5 ml) ) Trihydroborate (294.6 mg, 0.96 mmol) in N, N′-methanediylidenedipropan-2-amine (121 mg, 0.96 mmol) in N, N-dimethylformamide (1 ml). The solution was added and the reaction mixture was left at room temperature for 7 days. A further 133 mg of N, N′-methanediylidenedipropan-2-amine in 0.5 ml of N, N-dimethylformamide was added and the reaction continued for a further 7 days. The reaction was purified by column chromatography (5-10% methanol / ethyl acetate) and (E)-(2- (pyridin-3-ylimino) -1′-ammoniospiro [oxazolidine-5,3′-bicyclo [2.2.2] Octane] -1'-yl) trihydroborate was obtained (134.3 mg, 0.49 mmol, 51.3% yield).
¹ H NMR (500 MHz, MeOD) δ ppm 8.34-8.60 (m, 1 H), 8.16 (d, J = 4.27 Hz, 1 H), 7.59-7.96 (m, 1 H), 7.35 (dd, J = 8.24, 4.88 Hz, 1 H), 3.89 (br. S., 1 H), 3.65 (d, J = 9.46 Hz, 1 H), 3.16-3.23 (m, 1 H), 3.04-3.16 (m, 1 H), 2.85-3.04 (m, 2 H), 2.23 (br. S., 1 H), 1.74-1.97 (m, 4 H), 1.36-1.70 (m, 2 H). MS (LC / MS) RT = 0.65; [M + H-BH ₃ ] ⁺ = 259.21.

アセトン(５ml)中の(Ｅ)−(２−(ピリジン−３−イルイミノ)−１'−アンモニオスピロ[オキサゾリジン−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(１２７mg, ０.４７mmol)の懸濁液に、３Ｍ 塩酸(２ml, ６.００mmol)を加え、混合物を室温で２時間静置した。水およびクロロホルムを含む分液漏斗に、それを加えた。層を分離し、炭酸ナトリウム溶液で水層を塩基性にして、混合物をクロロホルムで再度抽出した。最後に、水相を酢酸エチルで洗浄した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(１％ 水酸化アンモニウム／９％ メタノール／９０％ ジクロロメタン)、ラセミの(Ｅ)−Ｎ−(１'−アザスピロ[オキサゾリジン−５,３'−ビシクロ[２.２.２]オクタン]−２−イリデン)ピリジン−３−アミンを白色の固体として得た(１９mg, ０.０７４mmol, 収率１５.８％)。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 8.39 (br. s., 1 H), 8.14 (d, J=4.27 Hz, 1 H), 7.72 (br. s., 1 H), 7.34 (dd, J=8.24, 4.88 Hz, 1 H), 3.89 (d, J=9.77 Hz, 1 H), 3.57 (d, J=10.38 Hz, 1 H), 3.14 - 3.27 (m, 1 H), 3.00 - 3.13 (m, 1 H), 2.70 - 3.00 (m, 4 H), 1.97 - 2.22 (m, 2 H), 1.54 - 1.84 (m, 3 H). MS (LC/MS) R.T. = 0.26; [M+H]⁺ = 259.16。 Step C: (E) -N- (1'-azaspiro [oxazolidine-5,3'-bicyclo [2.2.2] octane] -2-ylidene) pyridin-3-amine

(E)-(2- (Pyridin-3-ylimino) -1′-ammoniospiro [oxazolidine-5,3′-bicyclo [2.2.2] octane] -1′-yl in acetone (5 ml) ) To a suspension of trihydroborate (127 mg, 0.47 mmol) was added 3M hydrochloric acid (2 ml, 6.00 mmol) and the mixture was allowed to stand at room temperature for 2 hours. It was added to a separatory funnel containing water and chloroform. The layers were separated, the aqueous layer was basified with sodium carbonate solution and the mixture was extracted again with chloroform. Finally, the aqueous phase was washed with ethyl acetate. The organics were combined, dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (1% ammonium hydroxide / 9% methanol / 90% dichloromethane) and racemic (E) -N- (1′-azaspiro [oxazolidine-5,3′-bicyclo [2.2]. .2] Octane] -2-ylidene) pyridin-3-amine was obtained as a white solid (19 mg, 0.074 mmol, 15.8% yield).
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 8.39 (br. S., 1 H), 8.14 (d, J = 4.27 Hz, 1 H), 7.72 (br. S., 1 H), 7.34 (dd, J = 8.24, 4.88 Hz, 1 H), 3.89 (d, J = 9.77 Hz, 1 H), 3.57 (d, J = 10.38 Hz, 1 H), 3.14-3.27 (m, 1 H), 3.00-3.13 (m, 1 H), 2.70-3.00 (m, 4 H), 1.97-2.22 (m, 2 H), 1.54-1.84 (m, 3 H). MS (LC / MS) RT = 0.26; [M + H] ⁺ = 259.16.

Example 19
N- (pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

６−ブロモピリジン−２−アミン(２５３mg, １.４６mmol)、クロロホルム(２ml)、重炭酸ナトリウム(８５０mg, １０.１２mmol)および水(３ml)の混合物を集め、これに、クロロホルム(１ml)中のチオホスゲン(１９０mg, １.６５mmol)の溶液を加えた。反応混合物を室温で４時間撹拌した。反応混合物を分液漏斗に移し、酢酸エチルおよび水の層間に分配させた。有機層を塩水で洗浄し、硫酸マグネシウムで乾燥させ、濾過し、溶媒を蒸発させ、黄色の固体を得た。固体をカラムクロマトグラフィーによって精製し(５％ 酢酸エチル／ヘキサン)、２−ブロモ−６−イソチオシアナトピリジンを白色の固体として得た(２８１mg, １.３１mmol, 収率８９％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 7.50 - 7.62 (m, 1 H), 7.34 - 7.43 (m, 1 H), 6.91 - 7.11 (m, 1 H). MS (LC/MS) R.T. = 1.92; [M+H]⁺ = 216.86。 Step A: 2-Bromo-6-isothiocyanatopyridine

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 collected and added to chloroform (1 ml). A solution of thiophosgene (190 mg, 1.65 mmol) was added. 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 was evaporated to give a yellow solid. The solid was purified by column chromatography (5% ethyl acetate / hexane) to give 2-bromo-6-isothiocyanatopyridine as a white solid (281 mg, 1.31 mmol, 89% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.50-7.62 (m, 1 H), 7.34-7.43 (m, 1 H), 6.91-7.11 (m, 1 H). MS (LC / MS) RT = 1.92; [M + H] ⁺ = 216.86.

Ｎ,Ｎ−ジメチルホルムアミド(８ml)およびヒューニッヒ塩基(０.６ml, ３.４４mmol)中の、２−ブロモ−６−イソチオシアナトピリジン(２８１mg, １.３１mmol)の溶液に、(＋／−)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(３００mg, １.３１mmol)を加え、得られた混合物を７５℃で２.５時間加熱した。
MS (LC/MS) R.T. = 1.04; [M+H]⁺ = 373.01。 Step B: N- (6-Bromopyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

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), (+/−) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (300 mg, 1.31 mmol) was added and the resulting mixture was heated at 75 ° C. for 2.5 hours.
MS (LC / MS) RT = 1.04; [M + H] ⁺ = 373.01.

To this reaction mixture was added di-isopropyl-carbodiimide (523 mg, 4.14 mmol) and heating at 75 ° C. was continued for 2.25 hours. The mixture was cooled to room temperature over the weekend. The reaction mixture was concentrated in vacuo. The material was purified by column chromatography followed by preparative HPLC to give N- (6-bromopyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2. ] Octane] -2-amine was obtained as a yellow solid (291.5 mg, 0.86 mmol, 66.2% yield) (containing 2-amino-6-bromopyridine as an impurity).
MS (LC / MS) RT = 0.65; [M + H] ⁺ = 337.0.

メタノール(２０ml)中のＮ−(６−ブロモピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(２９１mg, ０.８６mmol)を、１０％ パラジウム／炭素(２３mg)で、Parr社製装置中で２時間水素化した。触媒を濾過によって除去し、濾液を真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(０.７％ 水酸化アンモニウム／６.３％ メタノール／９３％ クロロホルム)、ラセミのＮ−(ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(４１.３mg, ０.１６mmol, 収率１８.６％)。
¹H NMR (500 MHz, MeOD) δ ppm 8.13 - 8.33 (m, 1 H), 7.57 - 7.72 (m, 1 H), 6.83 - 7.04 (m, 2 H), 3.97 (d, J=10.07 Hz, 1 H), 3.66 (d, J=10.07 Hz, 1 H), 3.18 - 3.27 (m, 1 H), 3.03 - 3.14 (m, 1 H), 2.94 (t, J=7.63 Hz, 2 H), 2.70 - 2.90 (m, 2 H), 2.06 - 2.24 (m, 2 H), 1.57 - 1.87 (m, 3 H). MS (LC/MS) R.T. = 1.76; [M+H]⁺ = 259.25。 Step C: N- (Pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

N- (6-Bromopyridin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine (291 mg, in methanol (20 ml)) 0.86 mmol) was hydrogenated with 10% palladium / carbon (23 mg) in a Parr apparatus for 2 hours. The catalyst was removed by filtration and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (0.7% ammonium hydroxide / 6.3% methanol / 93% chloroform) and racemic N- (pyridin-2-yl) -4H-1′-azaspiro [oxazole-5 , 3′-bicyclo [2.2.2] octane] -2-amine was obtained (41.3 mg, 0.16 mmol, 18.6% yield).
¹ H NMR (500 MHz, MeOD) δ ppm 8.13-8.33 (m, 1 H), 7.57-7.72 (m, 1 H), 6.83-7.04 (m, 2 H), 3.97 (d, J = 10.07 Hz, 1 H), 3.66 (d, J = 10.07 Hz, 1 H), 3.18-3.27 (m, 1 H), 3.03-3.14 (m, 1 H), 2.94 (t, J = 7.63 Hz, 2 H), 2.70-2.90 (m, 2 H), 2.06-2.24 (m, 2 H), 1.57-1.87 (m, 3 H). MS (LC / MS) RT = 1.76; [M + H] ⁺ = 259.25.

Example 20
N- (4- (4-Methoxyphenyl) thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

アセトニトリル(３０ml)およびテトラヒドロフラン(５ml)中の、５−(４−メトキシフェニル)チアゾール−２−アミン(１.０７ｇ, ５.１９mmol)に、ジ(１Ｈ−イミダゾール−１−イル)メタンチオン(１.２０ｇ, ６.７４mmol)を加えた。反応物を６０℃で１８時間撹拌した。反応物を室温まで冷却し、沈殿物を濾過した。粉末を冷アセトニトリル(２×１５ml)で洗浄し、乾燥させ、Ｎ−(５−(４−メトキシフェニル)チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを橙色〜褐色の粉末として得た(０.５９ｇ, １.８６mmol, 収率３５.９％)。生成物をさらに特性決定することなく次の工程に直接用いた。 Step A: N- (5- (4-methoxyphenyl) thiazol-2-yl) -1H-imidazole-1-carbothioamide

5- (4-Methoxyphenyl) thiazol-2-amine (1.07 g, 5.19 mmol) in acetonitrile (30 ml) and tetrahydrofuran (5 ml) was added to di (1H-imidazol-1-yl) methanethione (1. 20 g, 6.74 mmol) was added. The reaction was stirred at 60 ° C. for 18 hours. The reaction was cooled to room temperature and the precipitate was filtered. The powder was washed with cold acetonitrile (2 × 15 ml) and dried to give N- (5- (4-methoxyphenyl) thiazol-2-yl) -1H-imidazole-1-carbothioamide as an orange-brown powder. (0.59 g, 1.86 mmol, yield 35.9%). The product was used directly in the next step without further characterization.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中のＮ−(４−(４−メトキシフェニル)チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(０.５７ｇ, １.８２mmol)に、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.３１ｇ, １.８２mmol)を加えた。反応物を６０℃で４時間撹拌した。反応物を冷却し、濃縮し、粗生成物を得た。粗製の物質をフラッシュクロマトグラフィーによって精製し(６０〜１００％ 酢酸エチル−ヘキサン)、最初のスポット／フラクション(ＴＬＣ)を生成物として得た。フラクションを合わせて、真空で濃縮し、(３−ヒドロキシ−３−((３−(４−(４−メトキシフェニル)チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを白色の粉末として得た(０.６ｇ, １.４３mmol, 収率７９％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 11.74 (s, 1 H), 7.91 (d, J=8.55 Hz, 2 H), 7.43 (s, 1 H), 6.97 (d, J=8.85 Hz, 2 H), 5.50 (s, 1 H), 3.62 - 3.98 (m, 5 H), 2.70 - 3.10 (m, 6 H), 2.13 (s, 1 H), 1.94 (s, 1 H), 1.66 - 1.91 (m, 2 H), 1.11 - 1.62 (m, 4 H). MS (LC/MS) R.T. = 3.54; [M+H]⁺ = 417.1。 Step B: (3-Hydroxy-3-((3- (4- (4-methoxyphenyl) thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2.2] octane-1- Yl) trihydroborate

N- (4- (4-methoxyphenyl) thiazol-2-yl) -1H-imidazole-1-carbothioamide (0.57 g, 1.82 mmol) in N, N-dimethylformamide (20 ml) was added to (3 -(Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (0.31 g, 1.82 mmol) was added. The reaction was stirred at 60 ° C. for 4 hours. The reaction was cooled and concentrated to give the crude product. The crude material was purified by flash chromatography (60-100% ethyl acetate-hexanes) to give the first spot / fraction (TLC) as product. The fractions were combined and concentrated in vacuo to give (3-hydroxy-3-((3- (4- (4-methoxyphenyl) thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2.2. .2] Octane-1-yl) trihydroborate was obtained as a white powder (0.6 g, 1.43 mmol, 79% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 11.74 (s, 1 H), 7.91 (d, J = 8.55 Hz, 2 H), 7.43 (s, 1 H), 6.97 (d, J = 8.85 Hz, 2 H), 5.50 (s, 1 H), 3.62-3.98 (m, 5 H), 2.70-3.10 (m, 6 H), 2.13 (s, 1 H), 1.94 (s, 1 H), 1.66-1.91 (m, 2 H), 1.11-1.62 (m, 4 H). MS (LC / MS) RT = 3.54; [M + H] ⁺ = 417.1.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(３−ヒドロキシ−３−((３−(４−(４−メトキシフェニル)チアゾール−２−イル)チオウレイド)メチル)−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(０.５９ｇ, １.４１mmol)に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.６６ml, ４.２３mmol)を加えた。反応物を７０℃で２４時間撹拌した。溶媒を真空で除去し、残渣をフラッシュクロマトグラフィーによって精製し(５０〜１００％ 酢酸エチル−ヘキサン)、最初の成分を生成物として集め、(２−(４−(４−メトキシフェニル)チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレートを白色の粉末として得た(０.４５ｇ, １.１７mmol, 収率８３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.65 (s, 1 H), 7.89 (d, J=7.32 Hz, 2 H), 7.15 - 7.43 (m, 1 H), 6.95 (d, J=8.85 Hz, 2 H), 3.64 - 3.93 (m, 5 H), 3.23 - 3.31 (m, J=1.53 Hz, 1 H), 3.09 - 3.21 (m, 1 H), 2.99 - 3.09 (m, 1 H), 2.79 - 2.97 (m, 3 H), 2.25 (s, 1 H), 1.96 - 2.16 (m, 1 H), 1.68 - 1.91 (m, 3 H), 1.45 (s, 3 H). MS (LC/MS) R.T. = 2.80; [M+H-BH₃]⁺ = 371.1。 Step C: (2- (4- (4-Methoxyphenyl) thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane] -1 '-Yl) trihydroborate

(3-Hydroxy-3-((3- (4- (4-methoxyphenyl) thiazol-2-yl) thioureido) methyl) -1-ammoniobicyclo [2] in N, N-dimethylformamide (20 ml). To [2.2] octan-1-yl) trihydroborate (0.59 g, 1.41 mmol) was added N, N′-diisopropylcarbodiimide (0.66 ml, 4.23 mmol). The reaction was stirred at 70 ° C. for 24 hours. The solvent is removed in vacuo, the residue is purified by flash chromatography (50-100% ethyl acetate-hexane), the first component is collected as product and (2- (4- (4-methoxyphenyl) thiazole-2 -Ylamino) -4H-1'-ammoniospiro [oxazol-5,3'-bicyclo [2.2.2] octane] -1'-yl) trihydroborate (0.45 g) as a white powder. , 1.17 mmol, 83% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.65 (s, 1 H), 7.89 (d, J = 7.32 Hz, 2 H), 7.15-7.43 (m, 1 H), 6.95 (d, J = 8.85 Hz, 2 H), 3.64-3.93 (m, 5 H), 3.23-3.31 (m, J = 1.53 Hz, 1 H), 3.09-3.21 (m, 1 H), 2.99-3.09 (m, 1 H), 2.79-2.97 (m, 3 H), 2.25 (s, 1 H), 1.96-2.16 (m, 1 H), 1.68-1.91 (m, 3 H), 1.45 (s, 3 H). MS (LC / MS) RT = 2.80 ; [M + H-BH 3] + = 371.1.

アセトン(９ml)中の(２−(４−(４−メトキシフェニル)チアゾール−２−イルアミノ)−４Ｈ−１'−アンモニオスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−１'−イル)トリヒドロボレート(０.４１ｇ, １.０７mmol)に、３Ｍ ＨＣｌ(０.３６ml, １.０７mmol)を加えた。反応物を室温で４時間撹拌した。ＴＬＣにより、反応の完了が示された(より低いスポット)。酢酸エチルを加え、水層を分離した。水層を１Ｎ 水酸化ナトリウムで中和した。生成物を酢酸エチル(２×４０ml)で抽出した。有機物を合わせて、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮し、ラセミのＮ−(４−(４−メトキシフェニル)チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.３ｇ, ０.７７mmol, 収率７２.１％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.56 (s, 1 H), 7.77 - 8.00 (m, J=8.55 Hz, 2 H), 7.27 (s, 1 H), 6.80 - 7.08 (m, 2 H), 3.87 (d, J=9.77 Hz, 1 H), 3.79 (s, 3 H), 3.61 (d, J=9.77 Hz, 1 H), 3.02 (s, 3 H), 2.60 - 2.92 (m, 4 H), 2.06 (s, 2 H), 1.82 - 2.00 (m, 1 H), 1.39 - 1.70 (m, 3 H). MS (LC/MS) R.T. = 1.95; [M+H]⁺ = 371.2。 Step D: N- (4- (4-Methoxyphenyl) thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(2- (4- (4-Methoxyphenyl) thiazol-2-ylamino) -4H-1′-ammoniospiro [oxazole-5,3′-bicyclo [2.2.2] octane in acetone (9 ml) ] -1'-yl) trihydroborate (0.41 g, 1.07 mmol) was added 3M HCl (0.36 ml, 1.07 mmol). The reaction was stirred at room temperature for 4 hours. TLC showed completion of reaction (lower spot). Ethyl acetate was added and the aqueous layer was separated. The aqueous layer was neutralized with 1N sodium hydroxide. The product was extracted with ethyl acetate (2 × 40 ml). The organics were combined, dried over magnesium sulfate, filtered, concentrated in vacuo, and racemic N- (4- (4-methoxyphenyl) thiazol-2-yl) -4H-1′-azaspiro [oxazole-5, 3′-Bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.3 g, 0.77 mmol, yield 72.1%).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.56 (s, 1 H), 7.77-8.00 (m, J = 8.55 Hz, 2 H), 7.27 (s, 1 H), 6.80-7.08 (m , 2 H), 3.87 (d, J = 9.77 Hz, 1 H), 3.79 (s, 3 H), 3.61 (d, J = 9.77 Hz, 1 H), 3.02 (s, 3 H), 2.60-2.92 (m, 4 H), 2.06 (s, 2 H), 1.82-2.00 (m, 1 H), 1.39-1.70 (m, 3 H). MS (LC / MS) RT = 1.95; [M + H] ⁺ = 371.2.

The enantiomers were separated using a Chiralpak AD-H (30 × 250 mm, 5 μm) column with a mobile phase consisting of 30% methanol in CO ₂ (0.1% DEA). The wavelength was set at 220 nm. The separated peak was concentrated in vacuo to give a white powder. The first peak of the column is (S) -N- (4- (4-methoxyphenyl) thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2. ] Octane] -2-amine (0.035 g, 0.09 mmol, yield 22.87%).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.56 (1 H, br.s.), 7.86 (2 H, d, J = 8.55 Hz), 7.25 (1 H, s), 6.93-6.96 ( 2 H, m), 3.86 (1 H, d, J = 9.77 Hz), 3.78 (3 H, s), 3.60 (1 H, d, J = 9.46 Hz), 3.01 (2 H, s), 2.72- 2.84 (2 H, m), 2.62-2.71 (2 H, m), 2.05 (1 H, br. S.), 1.91 (1 H, br. S.), 1.55-1.64 (2 H, m), 1.44-1.52 (1 H, m). MS (LC / MS) RT = 2.03; [M + H] ⁺ = 371.3.

The second peak is (R) -N- (4- (4-methoxyphenyl) thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2]. Octane] -2-amine (0.055 g, 0.15 mmol, yield 35.9%).
(21b, R-isomer): ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.58 (1 H, br. S.), 7.87 (2 H, d, J = 8.55 Hz), 7.26 (1 H, s), 6.92-6.97 (2 H, m), 3.86 (1 H, d, J = 9.77 Hz), 3.78 (3 H, s), 3.60 (1 H, d, J = 9.77 Hz), 3.01 (2 H, s), 2.73-2.85 (2 H, m), 2.63-2.71 (2 H, m), 2.05 (1 H, br. S.), 1.91 (1 H, br. S.), 1.54 -1.64 (2 H, m), 1.43-1.53 (1 H, m). MS (LC / MS) RT = 2.03; [M + H] ⁺ = 371.3.

Example 21
(R) -N- (2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazol-6-yl) acetamide

ジクロロメタン(１５０ml)中の、(３−(アミノメチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(１０ｇ, ４７.０mmol)に、炭酸ナトリウム(２００ml, ２００mmol)およびクロロ蟻酸ベンジル(９.５ml, ６６.５mmol)を加えた。反応混合物を室温で４０分間撹拌した。ジクロロメタンおよび水を加え、水層を分離し、ジクロロメタン(２×)で再度抽出した。有機層を合わせて、硫酸ナトリウムで乾燥させ、濾過し、濃縮した。残渣をフラッシュクロマトグラフィーによって精製した(１２〜１００％ 酢酸エチル−ヘキサン)。生成物のフラクションを合わせて、濃縮し、ラセミの(３−((ベンジルオキシカルボニルアミノ)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレートを透明な油状物として得た(３ｇ, ９.８６mmol, 収率２０.９６％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 7.30 - 7.43 (5 H, m), 5.27 (1 H, br. s.), 5.12 (2 H, s), 3.36 (2 H, d, J=6.04 Hz), 2.76 - 3.21 (6 H, m), 2.21 (1 H, br. s.), 1.97 (1 H, br. s.), 1.71 - 1.86 (2 H, m)。 Step A: (3-((Benzyloxycarbonylamino) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate

(3- (Aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate (10 g, 47.0 mmol) in dichloromethane (150 ml) was added to carbonic acid. Sodium (200 ml, 200 mmol) and benzyl chloroformate (9.5 ml, 66.5 mmol) were added. The reaction mixture was stirred at room temperature for 40 minutes. Dichloromethane and water were added and the aqueous layer was separated and extracted again with dichloromethane (2 ×). The organic layers were combined, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (12-100% ethyl acetate-hexane). The product fractions were combined, concentrated, and racemic (3-((benzyloxycarbonylamino) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydro. The borate was obtained as a clear oil (3 g, 9.86 mmol, 20.96% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.30-7.43 (5 H, m), 5.27 (1 H, br. S.), 5.12 (2 H, s), 3.36 (2 H, d, J = 6.04 Hz), 2.76-3.21 (6 H, m), 2.21 (1 H, br. S.), 1.97 (1 H, br. S.), 1.71-1.86 (2 H, m).

The enantiomers were separated on a mobile phase consisting of 20% acetonitrile / methanol (1: 1) in CO ₂ using a Chiralpak OJ-H (5 × 25) column. The wavelength was set at 210 nm. The first peak of the column is (R)-(3- (aminomethyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octan-1-yl) trihydroborate as a colorless oil. (37.67 g, 123 mmol).
Optical rotation: +28.2, c = 2.9 in chloroform.
The second peak of the column is (S)-(3-((benzyloxycarbonylamino) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane as a light amber oil. -1-yl) trihydroborate (46.82 g, 153 mmol).
Optical rotation: 27.4, c = 2.5 in chloroform.

アセトン(１２０ml)中の(Ｓ)−(３−((ベンジルオキシカルボニルアミノ)メチル)−３−ヒドロキシ−１−アンモニオビシクロ[２.２.２]オクタン−１−イル)トリヒドロボレート(２０.５ｇ, ６７mmol)の溶液を、氷浴で冷却した。３Ｍ 水性ＨＣｌ(１２０ml, ３６０mmol)を、２分かけて加えた。激しい発泡が観察された。１０分後、氷浴を除き、混合物を室温まで温めた。２０分後、それをメタノール(８００ml)で希釈し、窒素を吹き付けた。パラジウム／炭素(２ｇ, １.８８mmol)を加え、反応物に窒素を吹きつけ、水素のバルーンを取り付けた。反応混合物を室温で一夜撹拌した。それに窒素を吹きつけ、メタノールを用いて、セライトのパッドで濾過した。溶媒を蒸発させ、粗製の黄色の固体を得た。固体を水(２５ml)に溶解し、エタノール(４００ml)を加えた。すぐに白色の結晶が形成した。これを濾過によって集め、エタノールで、次にエーテルで洗浄した。白色の結晶性固体である(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩を得た(１０.７ｇ, ４６.７mmol, 収率６９.３％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 10.93 (1 H, br. s.), 8.24 (3 H, br. s.), 6.02 (1 H, s), 3.26 (1 H, d, J=13.43 Hz), 2.99 - 3.22 (5 H, m), 2.10 - 2.19 (2 H, m), 1.81 - 1.90 (1 H, m), 1.72 - 1.81 (1 H, m), 1.60 - 1.72 (1 H, m). 旋光度: [α]²⁰ _D = -50.9°(c = 6.4, 水)。 Step B: (S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride

(S)-(3-((Benzyloxycarbonylamino) methyl) -3-hydroxy-1-ammoniobicyclo [2.2.2] octane-1-yl) trihydroborate (20) in acetone (120 ml) 0.5 g, 67 mmol) was cooled in an ice bath. 3M aqueous HCl (120 ml, 360 mmol) was added over 2 minutes. Vigorous foaming was observed. After 10 minutes, the 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 sparged with nitrogen. Palladium / carbon (2 g, 1.88 mmol) was added, the reaction was sparged with nitrogen and a hydrogen balloon was attached. The reaction mixture was stirred overnight at room temperature. It was blown with nitrogen and filtered through a pad of celite with methanol. The solvent was evaporated to give a crude yellow solid. The solid was dissolved in water (25 ml) and ethanol (400 ml) was added. Soon white crystals formed. This was collected by filtration and washed with ethanol and then with ether. (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride, a white crystalline solid, was obtained (10.7 g, 46.7 mmol, 69.3% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 10.93 (1 H, br. S.), 8.24 (3 H, br. S.), 6.02 (1 H, s), 3.26 (1 H, d , J = 13.43 Hz), 2.99-3.22 (5 H, m), 2.10-2.19 (2 H, m), 1.81-1.90 (1 H, m), 1.72-1.81 (1 H, m), 1.60-1.72 (1 H, m). Optical rotation: [α] ²⁰ _D = -50.9 ° (c = 6.4, water).

アセトニトリル(１００ml)中のＮ−(２−アミノベンゾ[ｄ]チアゾール−６−イル)アセトアミド(４ｇ, １９.３mmol)に、ジ(１Ｈ−イミダゾール−１−イル)メタンチオン(３.４４ｇ, １９.３０mmol)を加えた。反応物を８０℃で一夜撹拌した。反応物を室温まで冷却し、沈殿物を濾過した。生成物であるＮ−(２−(１Ｈ−イミダゾール−１−カルボチオアミド)ベンゾ[ｄ]チアゾール−６−イル)アセトアミド(３.６ｇ, １１.３４mmol, 収率５８.８％)を、さらに精製することなく次の工程に直接用いた。 Step C: N- (2- (1H-imidazole-1-carbothioamido) benzo [d] thiazol-6-yl) acetamide

N- (2-aminobenzo [d] thiazol-6-yl) acetamide (4 g, 19.3 mmol) in acetonitrile (100 ml) was added to di (1H-imidazol-1-yl) methanethione (3.44 g, 19.30 mmol). ) Was added. The reaction was stirred at 80 ° C. overnight. The reaction was cooled to room temperature and the precipitate was filtered. The product, N- (2- (1H-imidazol-1-carbothioamido) benzo [d] thiazol-6-yl) acetamide (3.6 g, 11.34 mmol, 58.8% yield) was further purified. Used directly in the next step without.

Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中のＮ−(２−(１Ｈ−イミダゾール−１−カルボチオアミド)ベンゾ[ｄ]チアゾール−６−イル)アセトアミド(３００mg, ０.９５mmol)に、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２３８mg, １mmol)およびトリエチルアミン(０.３９ml, ２.８４mmol)を加えた。反応物を８０℃で３時間加熱した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.５９ml, ３.７８mmol)を反応混合物に加えた。混合物を８０℃でさらに２時間加熱した。反応物を冷却し、クロロホルムおよび水を混合物に加えた。有機層を真空で濃縮し、粗生成物を得た。粗製の物質を、フラッシュクロマトグラフィーによって精製した(２〜２０％ [１０％水酸化アンモニウム／メタノール]−クロロホルム)。生成物のフラクションをエーテルで磨砕し、(Ｒ)−Ｎ−(２−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)ベンゾ[ｄ]チアゾール−６−イル)アセトアミドを白色の固体として得た(１４４.５mg, ０.３９mmol, 収率４１.２％)。
¹H NMR (300 MHz, DMSO-d₆) δ ppm 9.98 (1 H, s), 8.93 (1 H, br. s.), 8.12 (1 H, d, J=1.83 Hz), 7.52 (1 H, d, J=8.42 Hz), 7.38 (1 H, dd, J=8.60, 2.01 Hz), 3.88 (1 H, d, J=9.88 Hz), 3.62 (1 H, d, J=9.88 Hz), 3.02 (2 H, s), 2.74 - 2.85 (2 H, m), 2.66 (2 H, t, J=7.68 Hz), 2.05 (4 H, s), 1.91 (1 H, br. s.), 1.41 - 1.64 (3 H, m). MS (LC/MS) R.T. = 1.55; [M+H]⁺ = 372.2。 Step D: (R) -N- (2- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) benzo [d] thiazole-6 Yl) acetamide

N- (2- (1H-imidazol-1-carbothioamido) benzo [d] thiazol-6-yl) acetamide (300 mg, 0.95 mmol) in N, N-dimethylformamide (10 ml) was added to (S)- 3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (238 mg, 1 mmol) and triethylamine (0.39 ml, 2.84 mmol) were added. The reaction was heated at 80 ° C. for 3 hours. N, N′-diisopropylcarbodiimide (0.59 ml, 3.78 mmol) was added to the reaction mixture. The mixture was heated at 80 ° C. for a further 2 hours. The reaction was cooled and chloroform and water were added to the mixture. The organic layer was concentrated in vacuo to give the crude product. The crude material was purified by flash chromatography (2-20% [10% ammonium hydroxide / methanol] -chloroform). The product fraction was triturated with ether and (R) -N- (2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) Benzo [d] thiazol-6-yl) acetamide was obtained as a white solid (144.5 mg, 0.39 mmol, 41.2% yield).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 9.98 (1 H, s), 8.93 (1 H, br. S.), 8.12 (1 H, d, J = 1.83 Hz), 7.52 (1 H , d, J = 8.42 Hz), 7.38 (1 H, dd, J = 8.60, 2.01 Hz), 3.88 (1 H, d, J = 9.88 Hz), 3.62 (1 H, d, J = 9.88 Hz), 3.02 (2 H, s), 2.74-2.85 (2 H, m), 2.66 (2 H, t, J = 7.68 Hz), 2.05 (4 H, s), 1.91 (1 H, br.s.), 1.41-1.64 (3 H, m). MS (LC / MS) RT = 1.55; [M + H] ⁺ = 372.2.

Example 22
(R) -N- (6- (Difluoromethoxy) benzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

酢酸(９０ml)中の４−(ジフルオロメトキシ)アニリン(９.５５ｇ, ６０mmol)に、チオシアン酸カリウム(ＫＳＣＮ)(１２.４１ml, ２４０mmol)を加えた。混合物を２０分間撹拌した(ＫＳＣＮが溶液に溶解した)。この混合物に、酢酸(４０ml)中の臭素(３.０８ml, ６０.０mmol)を２０分かけて滴下した。反応物を室温で一夜撹拌した。それを８００mlの氷水および２００mlの飽和水酸化アンモニウムの混合物に注いだ。生成物を酢酸エチル(５×)で抽出した。有機物を合わせて、塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、６−(ジフルオロメトキシ)ベンゾ[ｄ]チアゾール−２−アミンを黄色の固体として得た(１２.６ｇ, ５２.４mmol, 収率８７％)。 Step A: 6- (Difluoromethoxy) benzo [d] thiazol-2-amine

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 in the 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 (5x). The organics were combined, washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo to give 6- (difluoromethoxy) benzo [d] thiazol-2-amine as a yellow solid (12.6 g). , 52.4 mmol, 87% yield).

アセトニトリル(１５ml)中の６−(ジフルオロメトキシ)ベンゾ[ｄ]チアゾール−２−アミン(０.５ｇ, ２.３mmol)に、１,１'−チオカルボニルジイミダゾール(０.４９ｇ, ２.８mmol)を加えた。反応物を７０℃で一夜撹拌した。反応物を室温まで冷却し、沈殿物を濾過し、Ｎ−(６−(ジフルオロメトキシ)ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを黄色の固体として得た(５００mg, １.５３mmol, 収率６６.３％)。 Step B: N- (6- (Difluoromethoxy) benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide

6- (Difluoromethoxy) benzo [d] thiazol-2-amine (0.5 g, 2.3 mmol) in acetonitrile (15 ml) was added to 1,1′-thiocarbonyldiimidazole (0.49 g, 2.8 mmol). Was added. The reaction was stirred at 70 ° C. overnight. The reaction was cooled to room temperature and the precipitate was filtered to give N- (6- (difluoromethoxy) benzo [d] thiazol-2-yl) -1H-imidazole-1-carbothioamide as a yellow solid ( 500 mg, 1.53 mmol, 66.3% yield).

Ｎ,Ｎ−ジメチルホルムアミド(５ml)中のＮ−(６−(ジフルオロメトキシ)ベンゾ[ｄ]チアゾール−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(２８５mg, ０.８７mmol)の溶液に、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２００mg, ０.８７mmol)およびトリエチルアミン(０.４ml, ２.８７mmol)を加えた。反応物を７０℃で２時間加熱した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.４ml, ２.５７mmol)を、反応混合物に加えた。混合物を７０℃でさらに３時間加熱した。それを冷却し、トルエン／０.３Ｍ 水酸化ナトリウムに注いだ。生成物をトルエン(４×)およびクロロホルム(３×)で抽出した。有機物を合わせて、水(３×)で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、粗生成物を得た。粗製の物質を、フラッシュクロマトグラフィーによって精製し(２〜２０％ [１０％ 水酸化アンモニウム／メタノール]−クロロホルム)、(Ｒ)−Ｎ−(６−(ジフルオロメトキシ)ベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(１８６.２mg, ０.４９mmol, 収率５５.５％)。
融点 223〜5℃. ¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.99 (1 H, br. s.), 7.69 (1 H, d, J=2.75 Hz), 7.61 (1 H, d, J=8.55 Hz), 7.02 - 7.34 (2 H, m), 3.89 (1 H, d, J=10.07 Hz), 3.64 (1 H, d, J=9.77 Hz), 3.03 (2 H, d, J=2.44 Hz), 2.73 - 2.86 (2 H, m), 2.62 - 2.70 (2 H, m), 2.07 (1 H, br. s.), 1.92 (1 H, br. s.), 1.54 - 1.65 (2 H, m), 1.44 - 1.53 (1 H, m). MS (LC/MS) R.T. = 1.43; [M+H]⁺ = 381.1。 Step C: (R) -N- (6- (Difluoromethoxy) benzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

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), (S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (200 mg, 0.87 mmol) and triethylamine (0.4 ml, 2.87 mmol) were added. The reaction was heated at 70 ° C. for 2 hours. N, N′-diisopropylcarbodiimide (0.4 ml, 2.57 mmol) was added to the reaction mixture. The mixture was heated at 70 ° C. for an additional 3 hours. It was cooled and poured into toluene / 0.3M sodium hydroxide. The product was extracted with toluene (4x) and chloroform (3x). The organics were combined, washed with water (3x), dried over sodium sulfate, filtered and concentrated in vacuo to give the crude product. The crude material was purified by flash chromatography (2-20% [10% ammonium hydroxide / methanol] -chloroform) and (R) -N- (6- (difluoromethoxy) benzo [d] thiazole-2- Yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (186.2 mg, 0.49 mmol, yield 55). .5%).
Melting point 223-5 ° C. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.99 (1 H, br. S.), 7.69 (1 H, d, J = 2.75 Hz), 7.61 (1 H, d , J = 8.55 Hz), 7.02-7.34 (2 H, m), 3.89 (1 H, d, J = 10.07 Hz), 3.64 (1 H, d, J = 9.77 Hz), 3.03 (2 H, d, J = 2.44 Hz), 2.73-2.86 (2 H, m), 2.62-2.70 (2 H, m), 2.07 (1 H, br. S.), 1.92 (1 H, br. S.), 1.54- 1.65 (2 H, m), 1.44-1.53 (1 H, m). MS (LC / MS) RT = 1.43; [M + H] ⁺ = 381.1.

Example 23
(R) -N- (6-Methoxypyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１.８６ｇ, ７.９９mmol)の明橙色の溶液に、室温で、６−メトキシピリミジン−４−アミン(１ｇ, ８mmol)を加えた。橙色の溶液を室温で１８時間撹拌した。ＬＣ／ＭＳにより、望ましい生成物が主要なピークの１つとして示された。深橙色の溶液を濃縮し、残った残渣を濾過した。濾液をシリカゲルのクロマトグラフィーによって精製し(１０〜５０％ 酢酸エチル／ヘキサン)、４−イソチオシアナト−６−メトキシピリミジンを黄色の油状物として得た(０.７２ｇ, ４.３mmol, 収率５４％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.49 (1 H, d, J=5.79 Hz), 6.95 (1 H, d, J=5.79 Hz), 3.92 (3 H, s). MS (LC/MS) R.T. = 3.15; [M+H]⁺ = 168.1。 Step A: 4-isothiocyanato-6-methoxypyrimidine

To a light orange solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (1.86 g, 7.9 mmol) in dichloromethane at room temperature, 6-methoxypyrimidin-4-amine (1 g, 8 mmol). ) Was added. The orange solution was stirred at room temperature for 18 hours. 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 chromatography on silica gel (10-50% ethyl acetate / hexane) to give 4-isothiocyanato-6-methoxypyrimidine as a yellow oil (0.72 g, 4.3 mmol, 54% yield). .
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.49 (1 H, d, J = 5.79 Hz), 6.95 (1 H, d, J = 5.79 Hz), 3.92 (3 H, s). LC / MS) RT = 3.15; [M + H] ⁺ = 168.1.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(実施例２１の工程Ｂより)(０.３４ｇ, １.４９mmol)に、Ｃｓ_２ＣＯ_３(１.２２ｇ, ３.７４mmol)および４−イソチオシアナト−６−メトキシピリミジン(０.２５ｇ, １.５mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.７ml, ４.５mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜１５％ [９：１ メタノール：水酸化アンモニウム]／クロロホルム)、(Ｒ)−Ｎ−(６−メトキシピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の固体として得た(０.２１ｇ, ０.７２mmol, 収率４８.２％)。
融点 186〜8℃. ¹H NMR (400 MHz, MeOD) δ ppm 8.40 (1 H, s), 6.17 (1 H, br. s.), 3.92 - 4.04 (1 H, m), 3.89 (3 H, s), 3.68 (1 H, d, J=10.32 Hz), 3.12 - 3.23 (1 H, m), 2.98 - 3.12 (1 H, m), 2.67 - 2.97 (4 H, m), 2.11 (2 H, br. s.), 1.48 - 1.82 (3 H, m). MS (LC/MS) R.T. = 0.82; [M+H]⁺ = 290.3。 Step B: (R) -N- (6-Methoxypyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

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) _{cs 2 CO 3 (1.22g, 3.74mmol} ) and 4-isothiocyanato-6-methoxypyrimidine (0.25 g, 1.5 mmol) was added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.7 ml, 4.5 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-15% [9: 1 methanol: ammonium hydroxide] / chloroform), (R) -N- (6-methoxypyrimidin-4-yl) -4H- 1′-Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white solid (0.21 g, 0.72 mmol, 48.2% yield).
Melting point 186-8 ° C. ¹ H NMR (400 MHz, MeOD) δ ppm 8.40 (1 H, s), 6.17 (1 H, br. S.), 3.92-4.04 (1 H, m), 3.89 (3 H , s), 3.68 (1 H, d, J = 10.32 Hz), 3.12-3.23 (1 H, m), 2.98-3.12 (1 H, m), 2.67-2.97 (4 H, m), 2.11 (2 H, br. S.), 1.48-1.82 (3 H, m). MS (LC / MS) RT = 0.82; [M + H] ⁺ = 290.3.

Example 24
(R) -N- (6-Methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2] .2.2] Octane] -2-amine

ジオキサン(４０ml)中の５−メトキシピリジン−２−アミン(５ｇ, ４０mmol)に、エトキシカルボニル イソチオシアネート(５.２３ml, ４４.３mmol)を加えた。反応混合物を室温で一夜撹拌した。混合物を真空で濃縮し、５−メトキシピリジン−２−アミン エトキシカルボニル チオウレアを得た(１０.２８ｇ, ４０.３mmol, 収率１００％)。
¹H NMR (300 MHz, DMSO-d₆) δ ppm 12.03 (1 H, br. s.), 11.37 (1 H, br. s.), 8.53 (1 H, br. s.), 8.11 (1 H, d, J=2.93 Hz), 7.50 (1 H, dd, J=8.97, 3.11 Hz), 4.22 (2 H, q, J=7.07 Hz), 3.84 (3 H, s), 1.26 (3 H, t, J=7.14 Hz). MS (LC/MS) R.T. = 2.40; [M+H]⁺ = 256.1。 Step A: 5-methoxypyridin-2-amine ethoxycarbonyl thiourea

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 overnight at room temperature. The mixture was concentrated in vacuo to give 5-methoxypyridin-2-amine ethoxycarbonyl thiourea (10.28 g, 40.3 mmol, 100% yield).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 12.03 (1 H, br. S.), 11.37 (1 H, br. S.), 8.53 (1 H, br. S.), 8.11 (1 H, d, J = 2.93 Hz), 7.50 (1 H, dd, J = 8.97, 3.11 Hz), 4.22 (2 H, q, J = 7.07 Hz), 3.84 (3 H, s), 1.26 (3 H , t, J = 7.14 Hz). MS (LC / MS) RT = 2.40; [M + H] ⁺ = 256.1.

エタノール(５７ml)およびメタノール(５７ml)中の、５−メトキシピリジン−２−アミン エトキシカルボニル チオウレア(１０.２１ｇ, ４０mmol)に、ヒドロキシルアミン塩酸塩(１４ｇ, ２００mmol)およびヒューニッヒ塩基(２１ml, １２０mmol)を加えた。混合物を室温で２時間撹拌し、６０℃で４時間加熱した。反応物を室温まで冷却し、濾過し、固体を除去した。それを真空で濃縮し、クロロホルムに懸濁した。固体を濾過して取り、６−メトキシ−[１,２,４]トリアゾロ[１,５−ａ]ピリジン−２−アミンを得た(６.０５ｇ, ３３.２mmol, 収率８３％)。
¹H NMR (300 MHz, DMSO-d₆) δ ppm 8.30 (1 H, d, J=1.83 Hz), 7.24 - 7.32 (1 H, m), 7.16 - 7.23 (1 H, m), 5.82 (2 H, br. s.), 3.78 (3 H, s). MS (LC/MS) R.T. = 0.53; [M+H]⁺ = 165.2。 Step B: 6-Methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-amine

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). added. The mixture was stirred at room temperature for 2 hours and heated at 60 ° C. for 4 hours. The reaction was cooled to room temperature and filtered to remove solids. It was concentrated in vacuo and suspended in chloroform. The solid was filtered off to give 6-methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-amine (6.05 g, 33.2 mmol, 83% yield).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 8.30 (1 H, d, J = 1.83 Hz), 7.24-7.32 (1 H, m), 7.16-7.23 (1 H, m), 5.82 (2 H, br. S.), 3.78 (3 H, s). MS (LC / MS) RT = 0.53; [M + H] ⁺ = 165.2.

2 g of 6-methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-amine was purified by flash chromatography (2-20% [10% ammonium hydroxide / methanol] / chloroform. ), 6-methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-amine was obtained (1.35 g, 8.22 mmol, yield 67.5%).
¹ H NMR (300 MHz, DMSO-d ₆ ) δ ppm 8.29 (1 H, d, J = 2.20 Hz), 7.24-7.30 (1 H, m), 7.15-7.22 (1 H, m), 5.79 (2 H, s), 3.78 (3 H, s).

ジクロロメタン(１５ml)中の６−メトキシ−[１,２,４]トリアゾロ[１,５−ａ]ピリジン−２−アミン(０.３ｇ, １.８mmol)に、１,１'−チオカルボニル−ジ−２(１Ｈ)−ピリドン(０.５１ｇ, ２.２mmol)を加えた。反応物を室温で一夜撹拌し、真空で濃縮し、フラッシュクロマトグラフィーによって精製し、２−イソチオシアナト−６−メトキシ−[１,２,４]トリアゾロ[１,５−ａ]ピリジンを白色の固体として得た(１６６mg, ０.８mmol, 収率４４％)。
¹H NMR (300 MHz, CDCl₃) δ ppm 8.03 (1 H, d, J=2.20 Hz), 7.54 (1 H, d, J=9.88 Hz), 7.34 (1 H, dd, J=9.51, 2.56 Hz), 3.88 (3 H, s)。 Step C: 2-isothiocyanato-6-methoxy- [1,2,4] triazolo [1,5-a] pyridine

6-Methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-amine (0.3 g, 1.8 mmol) in dichloromethane (15 ml) was added to 1,1′-thiocarbonyl-di. -2 (1H) -pyridone (0.51 g, 2.2 mmol) was added. The reaction was stirred at room temperature overnight, concentrated in vacuo and purified by flash chromatography to give 2-isothiocyanato-6-methoxy- [1,2,4] triazolo [1,5-a] pyridine as a white solid. Obtained (166 mg, 0.8 mmol, 44% yield).
¹ H NMR (300 MHz, CDCl ₃ ) δ ppm 8.03 (1 H, d, J = 2.20 Hz), 7.54 (1 H, d, J = 9.88 Hz), 7.34 (1 H, dd, J = 9.51, 2.56 Hz), 3.88 (3 H, s).

Ｎ,Ｎ−ジメチルホルムアミド(５ml)中の２−イソチオシアナト−６−メトキシ−[１,２,４]トリアゾロ[１,５−ａ]ピリジン(１６０mg, ０.７８mmol)に、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１７８mg, ０.７８mmol)およびトリエチルアミン(０.３２ml, ２.３３mmol)を加えた。反応物を７０℃で１時間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.３６ml, ２.３３mmol)を反応混合物に加えた。混合物を７０℃で４時間加熱し、冷却し、水性重炭酸ナトリウム／クロロホルムに注いだ。生成物をクロロホルムで抽出した(３×)。合わせた有機物を水(３×)で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、フラッシュクロマトグラフィーによって精製し(２〜２０％ [１０％ 水酸化アンモニウム：メタノール]／クロロホルム)、(Ｒ)−Ｎ−(６−メトキシ−[１,２,４]トリアゾロ[１,５−ａ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ−[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(１１４.５mg, ０.３３mmol, 収率４２％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 8.51 (1 H, br. s.), 8.08 (1 H, d, J=2.44 Hz), 7.35 (1 H, d, J=9.77 Hz), 7.18 (1 H, dd, J=9.46, 2.44 Hz), 3.92 (1 H, d, J=8.85 Hz), 3.84 (3 H, s), 3.58 (1 H, d, J=8.85 Hz), 3.36 - 3.41 (1 H, m), 2.72 - 3.05 (4 H, m), 2.18 - 2.26 (1 H, m, J=13.26, 9.98, 3.66, 3.49, 3.49 Hz), 2.13 (1 H, br. s.), 1.79 (1 H, br. s.), 1.67 - 1.76 (1 H, m, J=13.96, 9.84, 4.27, 4.27 Hz), 1.45 - 1.63 (2 H, m). MS (LC/MS) R.T. = 0.86; [M+H]⁺ = 329.2。 Step D: (R) -N- (6-Methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′- Bicyclo [2.2.2] octane] -2-amine

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 to (S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (178 mg, 0.78 mmol) and triethylamine (0.32 ml, 2.33 mmol) were added. The reaction was stirred at 70 ° C. for 1 hour. N, N′-diisopropylcarbodiimide (0.36 ml, 2.33 mmol) was added to the reaction mixture. The mixture was heated at 70 ° C. for 4 hours, cooled and poured into aqueous sodium bicarbonate / chloroform. The product was extracted with chloroform (3x). The combined organics were washed with water (3 ×), dried over sodium sulfate, filtered, concentrated in vacuo and purified by flash chromatography (2-20% [10% ammonium hydroxide: methanol] / chloroform). , (R) -N- (6-methoxy- [1,2,4] triazolo [1,5-a] pyridin-2-yl) -4H-1′-azaspiro- [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine was obtained as a white powder (114.5 mg, 0.33 mmol, 42% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.51 (1 H, br.s.), 8.08 (1 H, d, J = 2.44 Hz), 7.35 (1 H, d, J = 9.77 Hz), 7.18 (1 H, dd, J = 9.46, 2.44 Hz), 3.92 (1 H, d, J = 8.85 Hz), 3.84 (3 H, s), 3.58 (1 H, d, J = 8.85 Hz), 3.36- 3.41 (1 H, m), 2.72-3.05 (4 H, m), 2.18-2.26 (1 H, m, J = 13.26, 9.98, 3.66, 3.49, 3.49 Hz), 2.13 (1 H, br. S. ), 1.79 (1 H, br. S.), 1.67-1.76 (1 H, m, J = 13.96, 9.84, 4.27, 4.27 Hz), 1.45-1.63 (2 H, m). MS (LC / MS) RT = 0.86; [M + H] ⁺ = 329.2.

Example 25
(R) -N- (5- (trifluoromethyl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

３Ｍ 水酸化ナトリウム(１８０ml, ５４０mmol)中の、５,６−ジアミノピリミジン−４−オール(１８ｇ, １４３mmol)の氷浴で冷却した溶液に、３,３−ジブロモ−１,１,１−トリフルオロプロパン−２−オン(２５.２ｇ, ９３mmol)を加えた。反応物を環境温度で３日間撹拌した。固体を濾過し、６０％ 硫酸(１４０ml)に溶解し、１３５℃で８時間撹拌した。反応物を冷却し、氷に注ぎ、１６時間置いた。濃水酸化アンモニウムで溶液をｐＨ ８まで中和し、酢酸エチル(５×１００ml)で抽出し、硫酸ナトリウムで乾燥させ、濾過し、濃縮した。固体残渣をベンゼン／ヘキサンから再結晶し、５−(トリフルオロメチル)ピラジン−２−アミンを得た(２.２８ｇ, １４mmol, 収率１５％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.32 (1 H, s), 7.99 (1 H, d, J=1.26 Hz), 5.02 (2 H, br. s.). MS (LC/MS) R.T. = 1.56; [M+H]⁺ = 164.03。 Step A: 5- (Trifluoromethyl) pyrazin-2-amine

To a cooled solution of 5,6-diaminopyrimidin-4-ol (18 g, 143 mmol) in 3M sodium hydroxide (180 ml, 540 mmol) in an ice bath was added 3,3-dibromo-1,1,1-trifluoro. Propan-2-one (25.2 g, 93 mmol) was added. The reaction was stirred at ambient temperature for 3 days. The solid was filtered, dissolved in 60% sulfuric acid (140 ml) and stirred at 135 ° C. for 8 hours. The reaction was cooled and poured onto ice and left for 16 hours. The solution was neutralized to pH 8 with concentrated ammonium hydroxide, extracted with ethyl acetate (5 × 100 ml), dried over sodium sulfate, filtered and concentrated. The solid residue was recrystallized from benzene / hexane to give 5- (trifluoromethyl) pyrazin-2-amine (2.28 g, 14 mmol, 15% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.32 (1 H, s), 7.99 (1 H, d, J = 1.26 Hz), 5.02 (2 H, br.s.). MS (LC / MS) RT = 1.56; [M + H] ⁺ = 164.03.

(Ｒ)−Ｎ−(５−(トリフルオロメチル)ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、５−(トリフルオロメチル)ピラジン−２−アミン(上記の工程Ａより)を出発物質として用いて製造した。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.08 (1 H, br. s.), 8.35 (1 H, s), 8.32 (1 H, s), 3.95 (1 H, d, J=9.57 Hz), 3.61 (1 H, d, J=9.57 Hz), 3.30 (1 H, dd, J=14.86, 1.76 Hz), 2.65 - 2.99 (5 H, m), 2.05 - 2.16 (2 H, m), 1.64 - 1.74 (1 H, m), 1.42 - 1.57 (2 H, m). MS (LC/MS) R.T. = 1.06; [M+H]⁺ = 328.30。 Step B: (R) -N- (5- (trifluoromethyl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (5- (trifluoromethyl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared according to the general procedure of Example 23, Steps AB, using 5- (trifluoromethyl) pyrazin-2-amine (from Step A above) as the starting material.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.08 (1 H, br.s.), 8.35 (1 H, s), 8.32 (1 H, s), 3.95 (1 H, d, J = 9.57 Hz ), 3.61 (1 H, d, J = 9.57 Hz), 3.30 (1 H, dd, J = 14.86, 1.76 Hz), 2.65-2.99 (5 H, m), 2.05-2.16 (2 H, m), 1.64-1.74 (1 H, m), 1.42-1.57 (2 H, m). MS (LC / MS) RT = 1.06; [M + H] ⁺ = 328.30.

Example 26
(R) -N- (6-Fluoro-1H-indazol-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

２,４−ジフルオロベンゾニトリル(１.２１ｇ, ８.７０mmol)に、ヒドラジン 一水和物(８.４６ml, １７４mmol)を加えた。混合物を５時間還流し、氷に注いだ。溶液を酢酸エチルで抽出し、硫酸マグネシウムで乾燥させ、濾過し、濃縮した。残渣をカラムクロマトグラフィーによって精製し(２５〜１００％ 酢酸エチル／ヘキサン)、６−フルオロ−１Ｈ−インダゾール−３−アミンを明黄色の粉末として得た(０.５ｇ, ３.３mmol, 収率３８％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 11.42 (s, 1 H), 7.70 (dd, J=8.55, 5.49 Hz, 1 H), 6.97 (dd, J=10.07, 1.83 Hz, 1 H), 6.72 - 6.79 (m, 1 H), 5.40 (s, 2 H). MS (LC/MS) R.T. = 0.61; [M+H]⁺ = 152.11。 Step A: 6-Fluoro-1H-indazole-3-amine

To 2,4-difluorobenzonitrile (1.21 g, 8.70 mmol) was added hydrazine monohydrate (8.46 ml, 174 mmol). The mixture was refluxed for 5 hours and poured into ice. The solution was extracted with ethyl acetate, dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography (25-100% ethyl acetate / hexane) to give 6-fluoro-1H-indazol-3-amine as a light yellow powder (0.5 g, 3.3 mmol, 38 yield). %).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 11.42 (s, 1 H), 7.70 (dd, J = 8.55, 5.49 Hz, 1 H), 6.97 (dd, J = 10.07, 1.83 Hz, 1 H) , 6.72-6.79 (m, 1 H), 5.40 (s, 2 H). MS (LC / MS) RT = 0.61; [M + H] ⁺ = 152.11.

ジクロロメタン(１５ml)中の６−フルオロ−１Ｈ−インダゾール−３−アミン(０.３２ｇ, ２.１mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.５３ｇ, ２.３０mmol)を加えた。反応物を５０℃で３時間撹拌した。反応物を室温まで冷却し、粗生成物をカラムクロマトグラフィーによって精製し(２５％ 酢酸エチル／ヘキサン)、６−フルオロ−３−イソチオシアナト−１Ｈ−インダゾールを明黄色の粉末として得た(０.３０ｇ, １.５３mmol, 収率７３.０％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 13.39 (s, 1 H), 7.78 (dd, J=8.85, 4.88 Hz, 1 H), 7.41 (dd, J=9.31, 1.68 Hz, 1 H), 7.14 (td, J=9.16, 1.83 Hz, 1 H). MS (LC/MS) R.T. = 3.69; [M+H]⁺ = 194.07。 Step B: 6-Fluoro-3-isothiocyanato-1H-indazole

6-Fluoro-1H-indazol-3-amine (0.32 g, 2.1 mmol) in dichloromethane (15 ml) was added to 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.53 g, 2 .30 mmol) was added. The reaction was stirred at 50 ° 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 give 6-fluoro-3-isothiocyanato-1H-indazole as a light yellow powder (0.30 g). 1.53 mmol, yield 73.0%).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 13.39 (s, 1 H), 7.78 (dd, J = 8.85, 4.88 Hz, 1 H), 7.41 (dd, J = 9.31, 1.68 Hz, 1 H) , 7.14 (td, J = 9.16, 1.83 Hz, 1 H). MS (LC / MS) RT = 3.69; [M + H] ⁺ = 194.07.

ＤＭＦ(１５ml)中の６−フルオロ−３−イソチオシアナト−１Ｈ−インダゾール(０.２０ｇ, １.０４mmol)に、トリエチルアミン(０.４３ml, ３.１１mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２６ｇ, １.１４mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.４８ml, ３.１１mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を環境温度まで冷却し、濃縮した。粗生成物をカラムクロマトグラフィーによって精製し(８５％ クロロホルム, １４％ メタノール, １％ 水酸化アンモニウム)、(Ｒ)−Ｎ−(６−フルオロ−１Ｈ−インダゾール−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.２２ｇ, ０.６６mmol, 収率６４％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 12.16 (s, 1 H), 8.00 (s, 1 H), 7.59 - 7.67 (m, 1 H), 7.09 (dd, J=9.77, 2.14 Hz, 1 H), 6.81 - 6.88 (m, 1 H), 3.81 (d, J=9.16 Hz, 1 H), 3.56 (d, J=8.85 Hz, 1 H), 3.00 (s, 2 H), 2.78 (s, 2 H), 2.67 (t, J=7.32 Hz, 2 H), 2.01 (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). MS (LC/MS) R.T. = 1.44; [M+H]⁺ = 316.16。 Step C: (R) -N- (6-Fluoro-1H-indazol-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

6-Fluoro-3-isothiocyanato-1H-indazole (0.20 g, 1.04 mmol) in DMF (15 ml) was added to triethylamine (0.43 ml, 3.11 mmol) and (S) -3- (aminomethyl) quinuclidine. -3-ol dihydrochloride (0.26 g, 1.14 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 heated at 70 ° 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), (R) -N- (6-fluoro-1H-indazol-3-yl) -4H-1 ′ -Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.22 g, 0.66 mmol, 64% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 12.16 (s, 1 H), 8.00 (s, 1 H), 7.59-7.67 (m, 1 H), 7.09 (dd, J = 9.77, 2.14 Hz, 1 H), 6.81-6.88 (m, 1 H), 3.81 (d, J = 9.16 Hz, 1 H), 3.56 (d, J = 8.85 Hz, 1 H), 3.00 (s, 2 H), 2.78 ( s, 2 H), 2.67 (t, J = 7.32 Hz, 2 H), 2.01 (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). MS (LC / MS) RT = 1.44; [M + H] ⁺ = 316.16.

Example 27
(R) -N- (furo [3,2-c] pyridin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

トルエン中の４−クロロフロ[３,２−ｃ]ピリジン(１ｇ, ６.５mmol)に、窒素下で、ラセミのＢＩＮＡＰ(０.２４３ｇ, ０.４mmol)、Ｐｄ_２(ｄｂａ)_３(０.１２ｇ, ０.１３mmol)およびナトリウム ｔｅｒｔ−ブトキシド(０.８８ｇ, ９.１mmol)を加えた。ベンゾフェノンイミン(１.３ml, ７.８１mmol)を加え、混合物を８０℃で３時間加熱し、室温まで冷却した。反応混合物をエーテルで希釈し、セライトで濾過し、エーテルで洗浄した。濾液を濃縮し、深橙色の残渣をメタノール(９０ml)に溶かし、ヒドロキシルアミン(１.２ml, １９.５mmol)で処理した。混合物を環境温度で１８時間撹拌し、濃縮した。残渣をカラムクロマトグラフィーによって精製し(９５〜１００％ 酢酸エチル／ヘキサン)、フロ[３,２−ｃ]ピリジン−４−アミンを深橙色の固体として得た(７７６mg, ５.７９mmol, 収率８９％)。
MS (LC/MS) R.T. = 0.51; [M+H]⁺ = 135.02。 Step A: furo [3,2-c] pyridin-4-amine

4-Chlorofuro [3,2-c] pyridine (1 g, 6.5 mmol) in toluene was added to racemic BINAP (0.243 g, 0.4 mmol), Pd ₂ (dba) ₃ (0.12 g) under nitrogen. , 0.13 mmol) and sodium tert-butoxide (0.88 g, 9.1 mmol). Benzophenone imine (1.3 ml, 7.81 mmol) was added and the mixture was heated at 80 ° C. for 3 hours 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 dissolved in methanol (90 ml) and treated with hydroxylamine (1.2 ml, 19.5 mmol). The mixture was stirred at ambient temperature for 18 hours and concentrated. The residue was purified by column chromatography (95-100% ethyl acetate / hexane) to give furo [3,2-c] pyridin-4-amine as a deep orange solid (776 mg, 5.79 mmol, 89 yield) %).
MS (LC / MS) RT = 0.51; [M + H] ⁺ = 135.02.

(Ｒ)−Ｎ−(フロ[３,２−ｃ]ピリジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、フロ[３,２−ｃ]ピリジン−４−アミン(上記の工程Ａ)を出発物質として用いて製造した。
¹H NMR (400 MHz, MeOD) δ ppm 8.08 (1 H, d, J=5.79 Hz), 7.69 (1 H, d, J=2.01 Hz), 6.99 - 7.13 (2 H, m), 4.00 (1 H, d, J=10.07 Hz), 3.69 (1 H, d, J=10.07 Hz), 3.19 - 3.26 (1 H, m), 3.05 - 3.13 (1 H, m), 2.93 (2 H, t, J=7.43 Hz), 2.73 - 2.87 (2 H, m), 2.08 - 2.24 (2 H, m), 1.52 - 1.82 (3 H, m). MS (LC/MS) R.T. = 0.68; [M+H]⁺ = 299.19。 Step B: (R) -N- (furo [3,2-c] pyridin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- 2-Amine

(R) -N- (furo [3,2-c] pyridin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Was prepared according to the general procedure of steps 23-B of Example 23 using furo [3,2-c] pyridin-4-amine (step A above) as a starting material.
¹ H NMR (400 MHz, MeOD) δ ppm 8.08 (1 H, d, J = 5.79 Hz), 7.69 (1 H, d, J = 2.01 Hz), 6.99-7.13 (2 H, m), 4.00 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J = 10.07 Hz), 3.19-3.26 (1 H, m), 3.05-3.13 (1 H, m), 2.93 (2 H, t, J = 7.43 Hz), 2.73-2.87 (2 H, m), 2.08-2.24 (2 H, m), 1.52-1.82 (3 H, m). MS (LC / MS) RT = 0.68; (M + H ] ⁺ = 299.19.

Example 28
(R) -N- (5-phenylpyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

エタノール(３ml)に溶解した、５−ブロモピリジン−３−アミン(２４８mg, １.４３mmol)、Ｐｄ(ＰＰｈ_３)_４(５０.４mg, ０.０４mmol)、トルエン(３ml)、炭酸ナトリウム(２Ｍ, ３ml, ６mmol)およびフェニルボロン酸(１９５mg, １.６０mmol)の混合物を、油浴中、９０℃で４時間加熱し、室温まで１６時間冷却した。反応混合物を分液漏斗に移し、酢酸エチルと水の層間に分配した。水相を酢酸エチルで１回以上洗浄し、合わせた有機相を塩水で洗浄し、硫酸マグネシウムで乾燥させ、濾過し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(８０％ 酢酸エチル／ヘキサン)、５−フェニルピリジン−３−アミンを白色の固体として得た(３１.９mg, ０.１９mmol, 収率１３％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 8.17 - 8.42 (m, 1 H), 8.02 - 8.20 (m, 1 H), 7.32 - 7.62 (m, 4 H), 7.25 (s, 1 H), 7.06 - 7.20 (m, 1 H). MS (LC/MS) R.T. = 0.91; [M+H]⁺ = 171.09。 Step A: 5-Phenylpyridin-3-amine

5-Bromopyridin-3-amine (248 mg, 1.43 mmol), Pd (PPh ₃ ) ₄ (50.4 mg, 0.04 mmol), toluene (3 ml), sodium carbonate (2M, dissolved in ethanol (3 ml). A mixture of 3 ml, 6 mmol) and phenylboronic acid (195 mg, 1.60 mmol) was heated in an oil bath at 90 ° C. for 4 hours and cooled 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 one more time with ethyl acetate and the combined organic phases were washed with brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (80% ethyl acetate / hexane) to give 5-phenylpyridin-3-amine as a white solid (31.9 mg, 0.19 mmol, 13% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.17-8.42 (m, 1 H), 8.02-8.20 (m, 1 H), 7.32-7.62 (m, 4 H), 7.25 (s, 1 H), 7.06-7.20 (m, 1 H). MS (LC / MS) RT = 0.91; [M + H] ⁺ = 171.09.

(Ｒ)−Ｎ−(５−フェニルピリジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、５−フェニルピリジン−３−アミン(上記の工程Ａより)を出発物質として用いて製造した。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 8.40 (br. s., 2 H), 7.66 (d, J=7.32 Hz, 2 H), 7.46 - 7.56 (m, 3 H), 7.43 (d, J=7.32 Hz, 1 H), 3.79 - 4.02 (m, 1 H), 3.51 - 3.68 (m, 1 H), 3.22 (d, J=14.95 Hz, 1 H), 3.02 - 3.15 (m, 1 H), 2.72 - 2.99 (m, 3 H), 2.14 (br. s., 2 H), 1.76 (dd, J=9.31, 4.12 Hz, 3 H), 1.12 - 1.35 (m, 1 H). MS (LC/MS) R.T. = 0.90; [M+H]⁺ = 335.17。 Step B: (R) -N- (5-Phenylpyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-phenylpyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared using 5-phenylpyridin-3-amine (from Step A above) as the starting material according to the general procedure of Steps AB.
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 8.40 (br. S., 2 H), 7.66 (d, J = 7.32 Hz, 2 H), 7.46-7.56 (m, 3 H), 7.43 ( d, J = 7.32 Hz, 1 H), 3.79-4.02 (m, 1 H), 3.51-3.68 (m, 1 H), 3.22 (d, J = 14.95 Hz, 1 H), 3.02-3.15 (m, 1 H), 2.72-2.99 (m, 3 H), 2.14 (br.s., 2 H), 1.76 (dd, J = 9.31, 4.12 Hz, 3 H), 1.12-1.35 (m, 1 H). MS (LC / MS) RT = 0.90; [M + H] ⁺ = 335.17.

(Ｒ)−Ｎ−(２−フェニルピリジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、２−ブロモピリジン−４−アミンから、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.38 (d, J=5.49 Hz, 1 H), 7.88 (d, J=7.93 Hz, 2 H), 7.71 - 7.84 (m, 1 H), 7.48 (d, J=7.63 Hz, 3 H), 7.19 - 7.36 (m, 1 H), 3.94 - 4.09 (m, 1 H), 3.61 - 3.79 (m, 1 H), 3.17 - 3.27 (m, 1 H), 3.00 - 3.14 (m, 1 H), 2.74 - 3.00 (m, 4 H), 2.05 - 2.23 (m, 2 H), 1.55 - 1.86 (m, 3 H). MS (LC/MS) R.T. = 0.86; [M+H]⁺ = 335.23。 Example 29
(R) -N- (2-Phenylpyridin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (2-phenylpyridin-4-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 2-bromo Prepared from pyridine-4-amine according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, MeOD) δ ppm 8.38 (d, J = 5.49 Hz, 1 H), 7.88 (d, J = 7.93 Hz, 2 H), 7.71-7.84 (m, 1 H), 7.48 (d , J = 7.63 Hz, 3 H), 7.19-7.36 (m, 1 H), 3.94-4.09 (m, 1 H), 3.61-3.79 (m, 1 H), 3.17-3.27 (m, 1 H), 3.00-3.14 (m, 1 H), 2.74-3.00 (m, 4 H), 2.05-2.23 (m, 2 H), 1.55-1.86 (m, 3 H). MS (LC / MS) RT = 0.86; [M + H] ⁺ = 335.23.

(Ｒ)−Ｎ−(６−フェニルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−ブロモピリジン−２−アミンから、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 7.91 (d, J=7.63 Hz, 2 H), 7.71 (t, J=7.78 Hz, 1 H), 7.32 - 7.55 (m, 5 H), 4.04 (d, J=10.07 Hz, 1 H), 3.72 (d, J=9.77 Hz, 1 H), 3.23 (d, J=1.22 Hz, 1 H), 3.12 (s, 1 H), 2.95 (s, 2 H), 2.84 (s, 2 H), 2.16 (br. s., 2 H), 1.58 - 1.85 (m, 3 H). MS (LC/MS) R.T. = 0.75; [M+H]⁺ = 335.23。 Example 30
(R) -N- (6-Phenylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Phenylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 6-bromo. Prepared from pyridin-2-amine according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 7.91 (d, J = 7.63 Hz, 2 H), 7.71 (t, J = 7.78 Hz, 1 H), 7.32-7.55 (m, 5 H), 4.04 (d, J = 10.07 Hz, 1 H), 3.72 (d, J = 9.77 Hz, 1 H), 3.23 (d, J = 1.22 Hz, 1 H), 3.12 (s, 1 H), 2.95 (s , 2 H), 2.84 (s, 2 H), 2.16 (br. S., 2 H), 1.58-1.85 (m, 3 H). MS (LC / MS) RT = 0.75; [M + H] ⁺ = 335.23.

Example 31
(R) -N- (6- (Difluoromethoxy) -1-methyl-1H-indazol-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

４−(ジフルオロメトキシ)−２−フルオロベンゾニトリル(１ｇ, ５.３mmol)に、メチルヒドラジン(４.９２ｇ, １０７mmol)を加えた。混合物を５０℃で５時間加熱し、室温まで冷却した。粗生成物をカラムクロマトグラフィーによって精製し(４０〜１００％ 酢酸エチル／ヘキサン)、６−(ジフルオロメトキシ)−１−メチル−１Ｈ−インダゾール−３−アミンを白色の粉末として得た(０.５ｇ, ２.３５mmol, 収率４４％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 7.70 (d, J=8.55 Hz, 1 H), 7.14 (s, 1 H), 6.73 (d, J=8.55 Hz, 1 H), 5.49 (s, 1 H), 3.70 (s, 3 H)。 Step A: 6- (Difluoromethoxy) -1-methyl-1H-indazole-3-amine

To 4- (difluoromethoxy) -2-fluorobenzonitrile (1 g, 5.3 mmol) was added methylhydrazine (4.92 g, 107 mmol). The mixture was heated at 50 ° C. for 5 hours and cooled to room temperature. The crude product was purified by column chromatography (40-100% ethyl acetate / hexane) to give 6- (difluoromethoxy) -1-methyl-1H-indazol-3-amine as a white powder (0.5 g , 2.35 mmol, 44% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 7.70 (d, J = 8.55 Hz, 1 H), 7.14 (s, 1 H), 6.73 (d, J = 8.55 Hz, 1 H), 5.49 ( s, 1 H), 3.70 (s, 3 H).

(Ｒ)−Ｎ−(６−(ジフルオロメトキシ)−１−メチル−１Ｈ−インダゾール−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、６−(ジフルオロメトキシ)−１−メチル−１Ｈ−インダゾール−３−アミン(工程Ａ)を出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 7.64 (d, J=8.55 Hz, 1 H), 7.28 (d, J=5.19 Hz, 1 H), 6.83 (dd, J=8.55, 1.83 Hz, 1 H), 3.88 (s, 3 H), 3.77 - 3.84 (m, 1 H), 3.51 - 3.64 (m, 1 H), 3.00 (s, 2 H), 2.72 - 2.84 (m, 2 H), 2.67 (t, J=7.48 Hz, 2 H), 2.02 (br. s., 1 H), 1.85 - 1.98 (m, 1 H), 1.59 (d, J=5.80 Hz, 2 H), 1.36 - 1.53 (m, 1 H), 1.09 (d, J=6.41 Hz, 1 H). MS (LC/MS) R.T. = 1.04; [M+H]⁺ = 378.19。 Step B: (R) -N- (6- (Difluoromethoxy) -1-methyl-1H-indazol-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2. 2] Octane] -2-amine

(R) -N- (6- (Difluoromethoxy) -1-methyl-1H-indazol-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-amine according to the general procedure of Example 23 Steps AB, using 6- (difluoromethoxy) -1-methyl-1H-indazol-3-amine (Step A) as starting material Manufactured.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 7.64 (d, J = 8.55 Hz, 1 H), 7.28 (d, J = 5.19 Hz, 1 H), 6.83 (dd, J = 8.55, 1.83 Hz , 1 H), 3.88 (s, 3 H), 3.77-3.84 (m, 1 H), 3.51-3.64 (m, 1 H), 3.00 (s, 2 H), 2.72-2.84 (m, 2 H) , 2.67 (t, J = 7.48 Hz, 2 H), 2.02 (br. S., 1 H), 1.85-1.98 (m, 1 H), 1.59 (d, J = 5.80 Hz, 2 H), 1.36- 1.53 (m, 1 H), 1.09 (d, J = 6.41 Hz, 1 H). MS (LC / MS) RT = 1.04; [M + H] ⁺ = 378.19.

Example 32
(R) -N- (5- (Difluoromethoxy) thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine

アセトニトリル(５００ml)中の２−ヒドロキシ−５−ニトロピリジン(７ｇ, ５０mmol)に、硫酸ナトリウム(１.５ｇ, １０.６mmol)および２,２−ジフルオロ−２−(フルオロスルホニル)酢酸(６.２ml, ６０mmol)を加え、反応物を室温で１６時間撹拌した。飽和水性重炭酸ナトリウムで反応を停止させ、アセトニトリルを真空で除去した。残った水性成分を酢酸エチルで抽出し、塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。薄褐色の油状固体をエーテル／ヘキサンで磨砕し、濾過し、濾液を濃縮し、２−(ジフルオロメトキシ)−５−ニトロピリジンを黄色の油状物として得た(４.７ｇ, ２４.７mmol, 収率４９％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.14 (d, J=2.76 Hz, 1 H), 8.68 (dd, J=9.03, 2.76 Hz, 1 H), 7.98 (s, 0.5 H), 7.62 (s, 0.5 H), 7.34 (d, J=9.03 Hz, 1 H)。 Step A: 2- (Difluoromethoxy) -5-nitropyridine

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 stirred at room temperature for 16 hours. The reaction was quenched with saturated aqueous sodium bicarbonate and 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 light brown oily solid was triturated with ether / hexanes, filtered, and the filtrate was concentrated to give 2- (difluoromethoxy) -5-nitropyridine as a yellow oil (4.7 g, 24.7 mmol, Yield 49%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.14 (d, J = 2.76 Hz, 1 H), 8.68 (dd, J = 9.03, 2.76 Hz, 1 H), 7.98 (s, 0.5 H), 7.62 (s, 0.5 H), 7.34 (d, J = 9.03 Hz, 1 H).

脱気したメタノール(１００ml)中の２−(ジフルオロメトキシ)−５−ニトロピリジン(４.７ｇ, ２４.７mmol)に、１０％ パラジウム／炭素(５００mg, ０.４７mmol)を加え、反応物を、大気圧で、１時間水素化した。これに酢酸(２.８３ml, ４９.４mmol)を加え、反応物をセライトで濾過し、真空で濃縮し、６−(ジフルオロメトキシ)ピリジン−３−アミンをオリーブグリーン色の液体として得た(６.３３ｇ, ２５.９mmol, 収率１０５％)。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 7.60 (d, J=2.76 Hz, 1 H), 7.37 (s, 0.5 H), 7.15 (dd, J=8.66, 2.89 Hz, 1 H), 7.00 (s, 0.5 H), 6.71 (d, J=8.78 Hz, 1 H)。 Step B: 6- (Difluoromethoxy) pyridin-3-amine

To 2- (difluoromethoxy) -5-nitropyridine (4.7 g, 24.7 mmol) in degassed methanol (100 ml) was added 10% palladium / 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 give 6- (difluoromethoxy) pyridin-3-amine as an olive green liquid (6 .33 g, 25.9 mmol, 105% yield).
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 7.60 (d, J = 2.76 Hz, 1 H), 7.37 (s, 0.5 H), 7.15 (dd, J = 8.66, 2.89 Hz, 1 H), 7.00 (s, 0.5 H), 6.71 (d, J = 8.78 Hz, 1 H).

氷浴中で冷却した酢酸(１０ml)に、チオシアン酸カリウム(３.１８ｇ, ３２.８mmol)および６−(ジフルオロメトキシ)ピリジン−３−アミン(１ｇ, ４.１mmol)を加えた。反応物の温度が＜０℃に至るまで、反応物を氷−塩浴中で冷却した。反応物の温度が＜０℃に維持される速度で、酢酸(３ml)中の臭素(０.６５ml, １２.７mmol)の溶液を２時間かけて滴下した。これにより非常に濃厚な混合物を得た。添加完了後、混合物を撹拌し、一夜かけてゆっくりと室温まで温めた。一夜撹拌した後、水(５ml)を加え、混合物を油浴中で８５℃に加熱した。この混合物を熱いままで濾過した。黄色のフィルターケーキを反応フラスコに戻し、さらに５mlの酢酸を加えた。混合物を再度８５℃に加熱し、熱いままで濾過した。合わせた濾液を氷浴中で冷却し、濃水酸化アンモニウムでｐＨ ８まで中和した。黄色の沈殿物が形成し、これを濾過によって集めた。この粗製の物質を、カラムクロマトグラフィーによって精製し(１２〜１００％ 酢酸エチル／ヘキサン)、５−(ジフルオロメトキシ)チアゾロ[５,４−ｂ]ピリジン−２−アミンを黄色の固体として得た(３２１mg, １.４８mmol, 収率３６.１％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.64 - 7.81 (m, 2 H), 6.92 (d, J=8.53 Hz, 1 H). MS (LC/MS) R.T. = 1.66; [M+H]⁺ = 218.10。 Step C: 5- (Difluoromethoxy) thiazolo [5,4-b] pyridin-2-amine

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 temperature of the reaction reached <0 ° 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 such that the temperature of the reaction was maintained at <0 ° C. This gave a very thick mixture. After the addition was complete, the mixture was stirred and allowed to warm slowly to room temperature overnight. After stirring overnight, water (5 ml) was added and the mixture was heated to 85 ° C. in an oil bath. The mixture was filtered hot. The yellow filter cake was returned to the reaction flask and an additional 5 ml of acetic acid was added. The mixture was heated again to 85 ° C. and filtered hot. The combined filtrate was cooled in an ice bath and neutralized to pH 8 with concentrated ammonium hydroxide. A yellow precipitate formed and was collected by filtration. This crude material was purified by column chromatography (12-100% ethyl acetate / hexane) to give 5- (difluoromethoxy) thiazolo [5,4-b] pyridin-2-amine as a yellow solid ( 321 mg, 1.48 mmol, yield 36.1%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.64-7.81 (m, 2 H), 6.92 (d, J = 8.53 Hz, 1 H). MS (LC / MS) RT = 1.66; (M + H] ⁺ = 218.10.

５−(ジフルオロメトキシ)チアゾロ[５,４−ｂ]ピリジン−２−アミン(３１０mg, １.４３mmol)およびジ(１Ｈ−イミダゾール−１−イル)メタンチオン(３１１mg, １.７５mmol)を、アセトニトリル(５ml)に溶解し、密封したバイアル中で、７０℃で１０時間加熱した。バイアルを冷凍庫中で１６時間保管した。固体を濾過によって集め、アセトニトリルで洗浄し、Ｎ−(５−(ジフルオロメトキシ)チアゾロ[５,４−ｂ]ピリジン−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミドを得た(２９６mg, ０.７２mmol, 収率５１％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.77 (d, J=4.77 Hz, 1 H), 8.06 (d, J=8.53 Hz, 1 H), 8.01 (s, 1 H), 7.72 (s, 1 H), 7.54 (s, 1 H), 6.94 - 7.13 (m, 1 H)。 Step D: N- (5- (Difluoromethoxy) thiazolo [5,4-b] pyridin-2-yl) -1H-imidazole-1-carbothioamide

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 added to acetonitrile (5 ml). ) And heated in a sealed vial at 70 ° C. for 10 hours. Vials were stored in a freezer for 16 hours. The solid was collected by filtration and washed with acetonitrile to give N- (5- (difluoromethoxy) thiazolo [5,4-b] pyridin-2-yl) -1H-imidazole-1-carbothioamide (296 mg, 0 0.72 mmol, 51% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.77 (d, J = 4.77 Hz, 1 H), 8.06 (d, J = 8.53 Hz, 1 H), 8.01 (s, 1 H), 7.72 ( s, 1 H), 7.54 (s, 1 H), 6.94-7.13 (m, 1 H).

Ｎ,Ｎ−ジメチルホルムアミド(４ml)中の、Ｎ−(５−(ジフルオロメトキシ)チアゾロ[５,４−ｂ]ピリジン−２−イル)−１Ｈ−イミダゾール−１−カルボチオアミド(２９６mg, ０.７２mmol)に、密封バイアル中で、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１７５mg, ０.７６mmol)およびトリエチルアミン(０.３ml, ２.２mmol)を加え、混合物を７０℃で一夜加熱した。これに、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(３５０μｌ, ２.２５mmol)を加え、反応物を７０℃で６時間加熱した。それを環境温度まで冷却し、水／クロロホルムに注ぎ、さらにクロロホルムで抽出し、真空で濃縮した。残渣をトルエンに溶かし、水で洗浄して、残ったＮ,Ｎ−ジメチルホルムアミドを除去した。残渣をカラムクロマトグラフィーによって精製し(２％〜２０％ (１０％ 水酸化アンモニウム／メタノール)／クロロホルム)、(Ｒ)−Ｎ−(５−(ジフルオロメトキシ)チアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを薄いクリーム色の固体として得た(１０４mg, ０.２７mmol, 収率３７％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.99 (d, J=8.78 Hz, 1 H), 7.68 (s, 1 H), 7.03 (d, J=8.53 Hz, 1 H), 3.87 (d, J=10.04 Hz, 1 H), 3.62 (d, J=10.04 Hz, 1 H), 3.03 (d, J=5.02 Hz, 2 H), 2.80 (d, J=9.03 Hz, 2 H), 2.65 (t, J=7.65 Hz, 2 H), 2.08 (br. s., 1 H), 1.83 - 1.99 (m, 1 H), 1.43 - 1.68 (m, 3 H). MS (LC/MS) R.T. = 1.73; [M+H]⁺ = 382.20。 Step E: (R) -N- (5- (difluoromethoxy) thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2 .2] Octane] -2-amine

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, (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (175 mg, 0.76 mmol) and triethylamine (0.3 ml, 2.2 mmol) are added and the mixture is added to 70 Heated at 0 C overnight. To this was added N, N′-diisopropylcarbodiimide (350 μl, 2.25 mmol) and the reaction was heated at 70 ° C. for 6 hours. It was cooled to ambient temperature, poured into water / chloroform, further extracted with chloroform and concentrated in vacuo. The residue was dissolved in toluene and washed with water to remove residual N, N-dimethylformamide. The residue was purified by column chromatography (2% -20% (10% ammonium hydroxide / methanol) / chloroform) and (R) -N- (5- (difluoromethoxy) thiazolo [5,4-b] pyridine- 2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a light cream solid (104 mg, 0.27 mmol, yield). Rate 37%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.99 (d, J = 8.78 Hz, 1 H), 7.68 (s, 1 H), 7.03 (d, J = 8.53 Hz, 1 H), 3.87 ( d, J = 10.04 Hz, 1 H), 3.62 (d, J = 10.04 Hz, 1 H), 3.03 (d, J = 5.02 Hz, 2 H), 2.80 (d, J = 9.03 Hz, 2 H), 2.65 (t, J = 7.65 Hz, 2 H), 2.08 (br. S., 1 H), 1.83-1.99 (m, 1 H), 1.43-1.68 (m, 3 H). MS (LC / MS) RT = 1.73; [M + H] ⁺ = 382.20.

(Ｒ)−Ｎ−(６−イソプロポキシベンゾ[ｄ]チアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−イソプロポキシアニリンから、実施例３２の工程Ｃ〜Ｅの一般的な手順に従って製造した。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.49 (d, J=8.78 Hz, 1 H), 7.38 (d, J=2.51 Hz, 1 H), 6.90 (dd, J=8.78, 2.51 Hz, 1 H), 4.50 - 4.68 (m, 1 H), 3.87 (d, J=10.04 Hz, 1 H), 3.62 (d, J=9.79 Hz, 1 H), 2.79 (d, J=9.03 Hz, 2 H), 2.66 (t, J=7.78 Hz, 2 H), 2.06 (br. s., 1 H), 1.85 - 1.96 (m, 1 H), 1.42 - 1.67 (m, 3 H), 1.17 - 1.34 (m, 6 H), 0.96 - 1.19 (m, 2 H)。 Example 33
(R) -N- (6-Isopropoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Isopropoxybenzo [d] thiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Was prepared from 4-isopropoxyaniline according to the general procedure of steps C-E of Example 32.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.49 (d, J = 8.78 Hz, 1 H), 7.38 (d, J = 2.51 Hz, 1 H), 6.90 (dd, J = 8.78, 2.51 Hz , 1 H), 4.50-4.68 (m, 1 H), 3.87 (d, J = 10.04 Hz, 1 H), 3.62 (d, J = 9.79 Hz, 1 H), 2.79 (d, J = 9.03 Hz, 2 H), 2.66 (t, J = 7.78 Hz, 2 H), 2.06 (br. S., 1 H), 1.85-1.96 (m, 1 H), 1.42-1.67 (m, 3 H), 1.17- 1.34 (m, 6 H), 0.96-1.19 (m, 2 H).

Example 34
(R) -N- (5- (pyrrolidin-1-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

５−ブロモピラジン−２−アミン(１.２ｇ, ６.９mmol)およびピロリジン(４ml, ４８mmol)の混合物を、１８０℃で２００Ｗで２時間マイクロ波照射した。反応物を１２５mlの酢酸エチルで希釈し、水(３×５０ml)および塩水(５０ml)で抽出した。それを硫酸ナトリウムで乾燥させ、濾過し、濃縮した。粗生成物をカラムクロマトグラフィーによって精製し(０〜３％ メタノール／塩化メチレン)、５−(ピロリジン−１−イル)ピラジン−２−アミンを得た(４９５mg, ３.mmol, 収率４３.７％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.52 (1 H, d, J=1.51 Hz), 7.36 (1 H, d, J=1.76 Hz), 5.21 (2 H, s), 3.24 - 3.29 (4 H, m), 1.90 (4 H, ddd, J=6.48, 3.53, 3.34 Hz). MS (LC/MS) R.T. = 0.52; [M+H]⁺ = 165.29。 Step A: 5- (Pyrrolidin-1-yl) pyrazin-2-amine

A mixture of 5-bromopyrazin-2-amine (1.2 g, 6.9 mmol) and pyrrolidine (4 ml, 48 mmol) was microwaved at 180 ° C. and 200 W for 2 hours. The reaction was diluted with 125 ml ethyl acetate and extracted with water (3 × 50 ml) and brine (50 ml). It was dried over sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (0-3% methanol / methylene chloride) to give 5- (pyrrolidin-1-yl) pyrazin-2-amine (495 mg, 3. mmol, yield 43.7). %).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.52 (1 H, d, J = 1.51 Hz), 7.36 (1 H, d, J = 1.76 Hz), 5.21 (2 H, s), 3.24- 3.29 (4 H, m), 1.90 (4 H, ddd, J = 6.48, 3.53, 3.34 Hz). MS (LC / MS) RT = 0.52; [M + H] ⁺ = 165.29.

(Ｒ)−Ｎ−(５−(ピロリジン−１−イル)ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、５−(ピロリジン−１−イル)ピラジン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, CDCl₃) δ ppm 8.51 (1 H, br. s.), 8.02 (1 H, s), 7.44 (1 H, d, J=0.92 Hz), 3.84 (1 H, d, J=9.16 Hz), 3.51 (1 H, d, J=8.85 Hz), 3.39 - 3.45 (4 H, m), 3.31 (1 H, dd, J=14.80, 1.07 Hz), 2.69 - 3.04 (5 H, m), 2.15 - 2.25 (1 H, m), 2.09 (1 H, br. s.), 1.98 - 2.02 (4 H + HOD, m), 1.64 - 1.74 (1 H, m), 1.42 - 1.61 (2 H, m). MS (LC/MS) R.T. = 0.76; [M+H]⁺ = 329.40。 Step B: (R) -N- (5- (pyrrolidin-1-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5- (pyrrolidin-1-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- The amine was prepared from 5- (pyrrolidin-1-yl) pyrazin-2-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.51 (1 H, br. S.), 8.02 (1 H, s), 7.44 (1 H, d, J = 0.92 Hz), 3.84 (1 H, d , J = 9.16 Hz), 3.51 (1 H, d, J = 8.85 Hz), 3.39-3.45 (4 H, m), 3.31 (1 H, dd, J = 14.80, 1.07 Hz), 2.69-3.04 (5 H, m), 2.15-2.25 (1 H, m), 2.09 (1 H, br.s.), 1.98-2.02 (4 H + HOD, m), 1.64-1.74 (1 H, m), 1.42- 1.61 (2 H, m). MS (LC / MS) RT = 0.76; [M + H] ⁺ = 329.40.

Example 35
(R) -1- (5- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) pyrazin-2-yl) pyrrolidin-2-one

５−ブロモピラジン−２−アミン(５ｇ, ２９mmol)、ピロリジン−２−オン(１１ml, １４４mmol)、ヨウ化銅(I)(１.１ｇ, ５.７５mmol)、炭酸カリウム(７.９４ｇ, ５７.５mmol)および(１Ｒ,２Ｒ)−シクロヘキサン−１,２−ジアミン(１.３８ml, １１.４９mmol)の混合物を、窒素下、ジオキサン(１００ml)中で１８時間還流した。冷却後、２００mlの酢酸エチルおよび２０mlのメタノールを反応物に加えた。これをセライトで濾過し、濃縮し、カラムクロマトグラフィーによって精製(０〜５％ メタノール／塩化メチレン)するために、硫酸ナトリウムに吸着させた。精製した生成物をエーテル／酢酸エチルから再結晶し、１−(５−アミノピラジン−２−イル)ピロリジン−２−オンを得た(１.５７ｇ, ８.８１mmol, 収率３０.７％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.74 (1 H, d, J=1.51 Hz), 7.69 (1 H, d, J=1.51 Hz), 6.20 (2 H, s), 3.84 (2 H, t, J=7.05 Hz), 2.49 (2 H, t, J=7.93 Hz), 2.04 (2 H, dq, J=7.68, 7.51 Hz). MS (LC/MS) R.T. = 0.48; [M+H]⁺ = 179.27。 Step A: 1- (5-Aminopyrazin-2-yl) pyrrolidin-2-one

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) were refluxed in dioxane (100 ml) for 18 hours under nitrogen. After cooling, 200 ml ethyl acetate and 20 ml methanol were added to the reaction. This was filtered through celite, concentrated and adsorbed onto sodium sulfate for purification by column chromatography (0-5% methanol / methylene chloride). The purified product was recrystallized from ether / ethyl acetate to give 1- (5-aminopyrazin-2-yl) pyrrolidin-2-one (1.57 g, 8.81 mmol, yield 30.7%). .
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.74 (1 H, d, J = 1.51 Hz), 7.69 (1 H, d, J = 1.51 Hz), 6.20 (2 H, s), 3.84 ( 2 H, t, J = 7.05 Hz), 2.49 (2 H, t, J = 7.93 Hz), 2.04 (2 H, dq, J = 7.68, 7.51 Hz). MS (LC / MS) RT = 0.48; [ M + H] ⁺ = 179.27.

(Ｒ)−１−(５−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)ピラジン−２−イル)ピロリジン−２−オンを、１−(５−アミノピラジン−２−イル)ピロリジン−２−オンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.20 (1 H, d, J=1.22 Hz), 8.91 (1 H, br. s.), 8.13 (1 H, s), 4.02 (2 H, t, J=7.02 Hz), 3.92 (1 H, d, J=9.46 Hz), 3.58 (1 H, d, J=9.16 Hz), 3.33 (1 H, dd, J=14.95, 1.53 Hz), 2.71 - 3.01 (5 H, m), 2.63 (2 H, t, J=8.09 Hz), 2.08 - 2.23 (4 H, m), 1.66 - 1.76 (1 H, m), 1.42 - 1.61 (2 H, m). MS (LC/MS) R.T. = 0.67; [M+H]⁺ = 343.30。 Step B: (R) -1- (5- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) pyrazin-2-yl) pyrrolidine- 2-on

(R) -1- (5- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) pyrazin-2-yl) pyrrolidin-2-one Was prepared from 1- (5-aminopyrazin-2-yl) pyrrolidin-2-one according to the general procedure of Example 23 Steps AB.
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.20 (1 H, d, J = 1.22 Hz), 8.91 (1 H, br. S.), 8.13 (1 H, s), 4.02 (2 H, t , J = 7.02 Hz), 3.92 (1 H, d, J = 9.46 Hz), 3.58 (1 H, d, J = 9.16 Hz), 3.33 (1 H, dd, J = 14.95, 1.53 Hz), 2.71- 3.01 (5 H, m), 2.63 (2 H, t, J = 8.09 Hz), 2.08-2.23 (4 H, m), 1.66-1.76 (1 H, m), 1.42-1.61 (2 H, m) MS (LC / MS) RT = 0.67; [M + H] ⁺ = 343.30.

Example 36
(R) -N- (5- (Pyridin-3-yl) pyrazin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- Amine

ピリジン−３−イルボロン酸(３０７mg, ２.５０mmol)、５−ブロモピラジン−２−アミン(３９１mg, ２.２５mmol)およびジクロロビス(トリフェニルホスフィン)−パラジウム(II)(８８mg, ０.１３mmol)を、脱気したジオキサン(１２ml)に加え、混合物を３０分間撹拌した。炭酸ナトリウム(７９５mg, ７.５０mmol)および脱気した水(８ml)を加え、反応物を、密封した反応バイアル中で、１００℃で８時間加熱した。反応物を環境温度で週末の間静置した。それを酢酸エチル(１００ml)で希釈し、塩水(３×２５ml)で抽出した。有機層を硫酸ナトリウムで乾燥させ、真空で濃縮した。粗生成物をカラムクロマトグラフィーによって精製し(０〜５％ メタノール／酢酸エチル)、５−(ピリジン−３−イル)ピラジン−２−アミンを得た(２３５mg, １.３７mmol, 収率５４.６％)。
¹H NMR (400 MHz, アセトン) δ ppm 9.13 (1 H, d, J=1.51 Hz), 8.54 (1 H, d, J=1.26 Hz), 8.50 (1 H, dd, J=4.78, 1.51 Hz), 8.22 - 8.28 (1 H, m), 8.07 (1 H, d, J=1.26 Hz), 7.39 (1 H, ddd, J=7.87, 4.72, 0.76 Hz), 6.05 (2 H, br. s.). MS (LC/MS) R.T. = 0.58; [M+H]⁺ = 173.20。 Step A: 5- (Pyridin-3-yl) pyrazin-2-amine

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. Degassed dioxane (12 ml) was added and the mixture was stirred for 30 minutes. Sodium carbonate (795 mg, 7.50 mmol) and degassed water (8 ml) were added and the reaction was heated in a sealed reaction vial at 100 ° C. for 8 hours. The reaction was left at ambient temperature for the weekend. It was diluted with ethyl acetate (100 ml) and extracted with brine (3 × 25 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude product was purified by column chromatography (0-5% methanol / ethyl acetate) to give 5- (pyridin-3-yl) pyrazin-2-amine (235 mg, 1.37 mmol, 54.6 yield). %).
¹ H NMR (400 MHz, acetone) δ ppm 9.13 (1 H, d, J = 1.51 Hz), 8.54 (1 H, d, J = 1.26 Hz), 8.50 (1 H, dd, J = 4.78, 1.51 Hz ), 8.22-8.28 (1 H, m), 8.07 (1 H, d, J = 1.26 Hz), 7.39 (1 H, ddd, J = 7.87, 4.72, 0.76 Hz), 6.05 (2 H, br. S .). MS (LC / MS) RT = 0.58; [M + H] ⁺ = 173.20.

(Ｒ)−Ｎ−(５−(ピリジン−３−イル)ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、５−(ピリジン−３−イル)ピラジン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.12 (1 H, dd, J=2.27, 0.76 Hz), 9.08 (1 H, br. s.), 8.59 (1 H, dd, J=4.78, 1.76 Hz), 8.55 (1 H, d, J=1.51 Hz), 8.45 (1 H, d, J=1.26 Hz), 8.25 (1 H, dt, J=7.99, 1.92 Hz), 7.38 (1 H, ddd, J=8.06, 4.78, 0.76 Hz), 3.97 (1 H, d, J=9.57 Hz), 3.63 (1 H, d, J=9.32 Hz), 3.36 (1 H, dd, J=14.86, 1.76 Hz), 2.69 - 3.06 (5 H, m), 2.14 - 2.24 (1 H, m), 2.12 (1 H, br. s.), 1.66 - 1.77 (1 H, m, J=13.94, 9.66, 4.31, 4.31 Hz), 1.44 - 1.62 (2 H, m). MS (LC/MS) R.T. = 0.65; [M+H]⁺ = 337.30。 Step B: (R) -N- (5- (Pyridin-3-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5- (Pyridin-3-yl) pyrazin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- The amine was prepared from 5- (pyridin-3-yl) pyrazin-2-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.12 (1 H, dd, J = 2.27, 0.76 Hz), 9.08 (1 H, br.s.), 8.59 (1 H, dd, J = 4.78, 1.76 Hz), 8.55 (1 H, d, J = 1.51 Hz), 8.45 (1 H, d, J = 1.26 Hz), 8.25 (1 H, dt, J = 7.99, 1.92 Hz), 7.38 (1 H, ddd , J = 8.06, 4.78, 0.76 Hz), 3.97 (1 H, d, J = 9.57 Hz), 3.63 (1 H, d, J = 9.32 Hz), 3.36 (1 H, dd, J = 14.86, 1.76 Hz ), 2.69-3.06 (5 H, m), 2.14-2.24 (1 H, m), 2.12 (1 H, br.s.), 1.66-1.77 (1 H, m, J = 13.94, 9.66, 4.31, 4.31 Hz), 1.44-1.62 (2 H, m). MS (LC / MS) RT = 0.65; [M + H] ⁺ = 337.30.

(Ｒ)−Ｎ−(５−(６−メトキシピリジン−３−イル)ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、５−ブロモピラジン−２−アミンから、実施例３６の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.04 (1 H, br. s.), 8.66 (1 H, d, J=2.01 Hz), 8.46 (1 H, d, J=1.51 Hz), 8.41 (1 H, d, J=1.01 Hz), 8.15 (1 H, dd, J=8.81, 2.52 Hz), 6.82 (1 H, d, J=8.31 Hz), 3.97 (3 H, s), 3.95 (1 H, d, J=9.57 Hz), 3.61 (1 H, d, J=9.32 Hz), 3.35 (1 H, dd, J=14.86, 1.51 Hz), 2.68 - 3.07 (5 H, m), 2.14 - 2.24 (1 H, m, J=13.27, 9.93, 3.53, 3.38, 3.38 Hz), 2.11 (1 H, br. s.), 1.67 - 1.77 (1 H, m), 1.45 - 1.61 (2 H, m). MS (LC/MS) R.T. = 0.81; [M+H]⁺ = 367.40。 Example 37
(R) -N- (5- (6-Methoxypyridin-3-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5- (6-Methoxypyridin-3-yl) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] 2-Amine was prepared from 5-bromopyrazin-2-amine according to the general procedure of Example 36, Steps AB.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.04 (1 H, br.s.), 8.66 (1 H, d, J = 2.01 Hz), 8.46 (1 H, d, J = 1.51 Hz), 8.41 (1 H, d, J = 1.01 Hz), 8.15 (1 H, dd, J = 8.81, 2.52 Hz), 6.82 (1 H, d, J = 8.31 Hz), 3.97 (3 H, s), 3.95 ( 1 H, d, J = 9.57 Hz), 3.61 (1 H, d, J = 9.32 Hz), 3.35 (1 H, dd, J = 14.86, 1.51 Hz), 2.68-3.07 (5 H, m), 2.14 -2.24 (1 H, m, J = 13.27, 9.93, 3.53, 3.38, 3.38 Hz), 2.11 (1 H, br. S.), 1.67-1.77 (1 H, m), 1.45-1.61 (2 H, m). MS (LC / MS) RT = 0.81; [M + H] ⁺ = 367.40.

Example 38
(R) -N- (6-Methoxyquinoxalin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

トルエン中５０％溶液の２−オキソ酢酸エチル(１８.４７ml, ９３mmol)を、エタノール(１００ml)中の４−メトキシベンゼン−１,２−ジアミン(１０.７３ｇ, ７８mmol)の溶液に環境温度で加え、反応物を２時間還流した。反応物を真空で濃縮し、エタノールから結晶化し、６−メトキシキノキサリン−２(１Ｈ)−オンおよび７−メトキシキノキサリン−２(１Ｈ)−オンの混合物を得た(５.７３ｇ, ３２.５０mmol, 収率４２％)。
MS (LC/MS) R.T. = 0.68; [M+H]⁺ = 177.10。 Step A: 7-methoxyquinoxalin-2 (1H) -one and 6-methoxyquinoxalin-2 (1H) -one

A 50% solution of ethyl 2-oxoacetate (18.47 ml, 93 mmol) in toluene is added to a solution of 4-methoxybenzene-1,2-diamine (10.73 g, 78 mmol) in ethanol (100 ml) at ambient temperature. The reaction was refluxed for 2 hours. The reaction was concentrated in vacuo and crystallized from ethanol to give 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) RT = 0.68; [M + H] ⁺ = 177.10.

６−メトキシキノキサリン−２(１Ｈ)−オンおよび７−メトキシキノキサリン−２(１Ｈ)−オン(５.６７ｇ, ３２.２０mmol)の混合物を、オキシ塩化リン(１２０ml)中で、１時間還流した。反応物を濃縮し、氷を添加することによって反応停止させ、炭酸ナトリウムで塩基性にして、酢酸エチル(３×２００ml)で抽出した。有機層を合わせて、真空で濃縮した。粗生成物を硫酸ナトリウムに吸着させ、カラムクロマトグラフィーによって精製し(０〜５％ 酢酸エチル／ヘキサン)、２−クロロ−６−メトキシキノキサリンを得た(２.２１ｇ, １１.３６mmol, 収率３５％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.69 (s, 1 H), 7.88 (d, J=9.32 Hz, 1 H), 7.43 (dd, J=9.32, 2.77 Hz, 1 H), 7.37 (d, J=2.77 Hz, 1 H), 3.95 (s, 3 H)。 Step B: 2-Chloro-6-methoxyquinoxaline

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 hour. The reaction was concentrated and quenched by the addition of ice, basified with sodium carbonate and extracted with ethyl acetate (3 × 200 ml). The organic layers were combined and concentrated in vacuo. The crude product was adsorbed on sodium sulfate and purified by column chromatography (0-5% ethyl acetate / hexane) to give 2-chloro-6-methoxyquinoxaline (2.21 g, 11.36 mmol, yield 35). %).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.69 (s, 1 H), 7.88 (d, J = 9.32 Hz, 1 H), 7.43 (dd, J = 9.32, 2.77 Hz, 1 H), 7.37 ( d, J = 2.77 Hz, 1 H), 3.95 (s, 3 H).

２−クロロ−６−メトキシキノキサリン(０.９３ｇ, ４.７７mmol)および(２,４−ジメトキシフェニル)メタンアミン(２.２ml, １４.６４mmol)を、ジメチルスルホキシド(５ml)中で、１５０℃で３０分間マイクロ波照射した。反応物を酢酸エチル(２５０ml)で希釈し、塩水(３×１００ml)で抽出した。粗生成物をカラムクロマトグラフィーによって精製し(２０〜８０％ 酢酸エチル／ヘキサン)、Ｎ−(２,４−ジメトキシベンジル)−６−メトキシキノキサリン−２−アミンを得た(１.４６ｇ, 収率８７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.13 (1 H, s), 7.59 - 7.63 (1 H, m), 7.30 (1 H, d, J=8.31 Hz), 7.21 - 7.24 (2 H, m), 6.47 (1 H, d, J=2.27 Hz), 6.42 (1 H, dd, J=8.31, 2.27 Hz), 5.10 (1 H, t, J=5.92 Hz), 4.61 (2 H, d, J=5.79 Hz), 3.88 (3 H, s), 3.84 (3 H, s), 3.78 (3 H, s). MS (LC/MS) R.T. = 1.95; [M+H]⁺ = 326.23。 Step C: N- (2,4-dimethoxybenzyl) -6-methoxyquinoxalin-2-amine

2-Chloro-6-methoxyquinoxaline (0.93 g, 4.77 mmol) and (2,4-dimethoxyphenyl) methanamine (2.2 ml, 14.64 mmol) were added in dimethyl sulfoxide (5 ml) at 150 ° C. for 30 minutes. Microwave irradiation for minutes. The reaction was diluted with ethyl acetate (250 ml) and extracted with brine (3 × 100 ml). The crude product was purified by column chromatography (20-80% ethyl acetate / hexane) to give N- (2,4-dimethoxybenzyl) -6-methoxyquinoxalin-2-amine (1.46 g, yield). 87%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.13 (1 H, s), 7.59-7.63 (1 H, m), 7.30 (1 H, d, J = 8.31 Hz), 7.21-7.24 (2 H, m), 6.47 (1 H, d, J = 2.27 Hz), 6.42 (1 H, dd, J = 8.31, 2.27 Hz), 5.10 (1 H, t, J = 5.92 Hz), 4.61 (2 H, d , J = 5.79 Hz), 3.88 (3 H, s), 3.84 (3 H, s), 3.78 (3 H, s). MS (LC / MS) RT = 1.95; [M + H] ⁺ = 326.23.

Ｎ−(２,４−ジメトキシベンジル)−６−メトキシキノキサリン−２−アミン(２.８ｇ, ８.６１mmol)を、ＴＦＡ(１０ml, １３０mmol)およびジクロロメタン(１０ml)中で、環境温度で３０分間撹拌した。溶媒を真空で除去した。飽和水性炭酸水素ナトリウム(２００ml)を赤色の残渣に加えると、黄色の固体が沈殿した。混合物を大量のジクロロメタンで抽出した。有機層を硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、６−メトキシキノキサリン−２−アミンを得た(１.５０ｇ, ８.５６mmol, 収率９９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.27 (1 H, s), 7.54 - 7.58 (1 H, m), 7.25 - 7.29 (2 H, m), 4.71 (2 H, br. s.), 3.90 (3 H, s). MS (LC/MS) R.T. = 0.86; [M+H]⁺ = 176.23。 Step D: 6-Methoxyquinoxalin-2-amine

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 minutes. did. The solvent was removed in vacuo. Saturated aqueous sodium bicarbonate (200 ml) was added to the red residue and a yellow solid precipitated. The mixture was extracted with a large amount of dichloromethane. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give 6-methoxyquinoxalin-2-amine (1.50 g, 8.56 mmol, 99% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.27 (1 H, s), 7.54-7.58 (1 H, m), 7.25-7.29 (2 H, m), 4.71 (2 H, br. S.) 3.90 (3 H, s). MS (LC / MS) RT = 0.86; [M + H] ⁺ = 176.23.

(Ｒ)−Ｎ−(６−メトキシキノキサリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−メトキシキノキサリン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.61 (1 H, br. s.), 8.57 (1 H, s), 7.62 (1 H, d, J=9.06 Hz), 7.23 - 7.32 (2 H, m), 4.01 (1 H, d, J=9.32 Hz), 3.90 (3 H, s), 3.66 (1 H, d, J=9.32 Hz), 3.37 (1 H, dd, J=14.86, 1.51 Hz), 2.68 - 3.08 (5 H, m), 2.15 - 2.25 (1 H, m), 2.10 - 2.14 (1 H, m), 1.67 - 1.77 (1 H, m), 1.42 - 1.63 (2 H, m). MS (LC/MS) R.T. = 0.81; [M+H]⁺ = 340.30。 Step E: (R) -N- (6-Methoxyquinoxalin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Methoxyquinoxalin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 6-methoxy. Prepared from quinoxalin-2-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.61 (1 H, br. S.), 8.57 (1 H, s), 7.62 (1 H, d, J = 9.06 Hz), 7.23-7.32 (2 H , m), 4.01 (1 H, d, J = 9.32 Hz), 3.90 (3 H, s), 3.66 (1 H, d, J = 9.32 Hz), 3.37 (1 H, dd, J = 14.86, 1.51 Hz), 2.68-3.08 (5 H, m), 2.15-2.25 (1 H, m), 2.10-2.14 (1 H, m), 1.67-1.77 (1 H, m), 1.42-1.63 (2 H, m). MS (LC / MS) RT = 0.81; [M + H] ⁺ = 340.30.

Example 39
(R) -N- (5- (Difluoromethoxy) pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)および水(０.２ml)中の、２−クロロピリミジン−５−オール(１ｇ, ７.６６mmol)および２−クロロ−２,２−ジフルオロ酢酸ナトリウム(３.５０ｇ, ２２.９８mmol)を、９０℃で２４時間加熱し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(５〜３０％ 酢酸エチル／ヘキサン)、２−クロロ−５−(ジフルオロメトキシ)ピリミジンを薄黄色の油状物として得た(５４９mg, ３.０４mmol, 収率３９.７％)。
MS (LC/MS) R.T. = 1.32; [M+H]⁺ = 181.14。 Step A: 2-Chloro-5- (difluoromethoxy) pyrimidine

2-chloropyrimidin-5-ol (1 g, 7.66 mmol) and sodium 2-chloro-2,2-difluoroacetate (3.50 g) in N, N-dimethylformamide (20 ml) and water (0.2 ml). , 22.98 mmol) was heated at 90 ° C. for 24 h and concentrated in vacuo. The residue was purified by column chromatography (5-30% ethyl acetate / hexane) to give 2-chloro-5- (difluoromethoxy) pyrimidine as a pale yellow oil (549 mg, 3.04 mmol, yield 39. 7%).
MS (LC / MS) RT = 1.32; [M + H] ⁺ = 181.14.

(Ｒ)−Ｎ−(５−(ジフルオロメトキシ)ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、２−クロロ−５−(ジフルオロメトキシ)ピリミジンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeO-d₄) δ ppm 8.46 (2 H, s), 6.85 (1 H, t), 4.02 (1 H, d, J=10.07 Hz), 3.73 (1 H, d, J=10.32 Hz), 3.28 (1 H, d, J=1.01 Hz), 3.16 (1 H, d), 2.81 - 3.05 (4 H, m), 2.08 - 2.25 (2 H, m), 1.46 - 1.89 (3 H, m). MS (LC/MS) R.T. = 0.53; [M+H]⁺ = 326.30。 Step B: (R) -N- (5- (Difluoromethoxy) pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (5- (difluoromethoxy) pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared from 2-chloro-5- (difluoromethoxy) pyrimidine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, MeO-d ₄ ) δ ppm 8.46 (2 H, s), 6.85 (1 H, t), 4.02 (1 H, d, J = 10.07 Hz), 3.73 (1 H, d, J = 10.32 Hz), 3.28 (1 H, d, J = 1.01 Hz), 3.16 (1 H, d), 2.81-3.05 (4 H, m), 2.08-2.25 (2 H, m), 1.46-1.89 (3 H, m). MS (LC / MS) RT = 0.53; [M + H] ⁺ = 326.30.

Example 40
(R) -N- (4,5-Dimethylpyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

メタノール中２Ｍ アンモニア(１００ml)中の、４−クロロ−５,６−ジメチルピリミジン−２−アミン(０.３５ｇ, ２.２２mmol)の溶液に、窒素を吹きつけ、パラジウム／炭素(０.０３５ｇ, ０.３３mmol)を加え、窒素を吹きつけ、反応物を、１atmで、環境温度で１８時間水素化した。反応混合物に窒素を吹きつけ、セライトで濾過し、セライトのパッドをメタノールで洗浄した。濾液を真空で乾固するまで蒸発させ、４,５−ジメチルピリミジン−２−アミンを得た(０.３５ｇ, ２.５６mmol, 収率９０％)。これをさらに精製することなく用いた。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.89 (1 H, s), 6.19 (2 H, s), 2.18 (3 H, s), 1.99 (3 H, s). MS (LC/MS) R.T. = 0.56; [M+H]⁺ = 124.20。 Step A: 4,5-dimethylpyrimidin-2-amine

A solution of 4-chloro-5,6-dimethylpyrimidin-2-amine (0.35 g, 2.22 mmol) in 2M ammonia in methanol (100 ml) was blown with nitrogen to produce palladium / carbon (0.035 g, 0.33 mmol) was added, nitrogen was blown, and the reaction was hydrogenated at 1 atm and ambient temperature for 18 hours. The reaction mixture was sparged with nitrogen, filtered through celite, and the celite pad was washed with methanol. The filtrate was evaporated to dryness in vacuo to give 4,5-dimethylpyrimidin-2-amine (0.35 g, 2.56 mmol, 90% yield). This was used without further purification.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.89 (1 H, s), 6.19 (2 H, s), 2.18 (3 H, s), 1.99 (3 H, s). MS (LC / MS) RT = 0.56; [M + H] ⁺ = 124.20.

(Ｒ)−Ｎ−(４,５−ジメチルピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４,５−ジメチルピリミジン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.22 (1 H, s), 4.01 (1 H, d, J=10.07 Hz), 3.73 (1 H, d, J=10.32 Hz), 3.40 (1 H, d), 3.25 (1 H, d), 2.91 - 3.12 (4 H, m), 2.41 (3 H, s), 2.09 - 2.28 (5 H, m), 1.62 - 1.97 (3 H, m). MS (LC/MS) R.T. = 0.47; [M+H]⁺ = 288.31。 Step B: (R) -N- (4,5-Dimethylpyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4,5-dimethylpyrimidin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 4 , 5-dimethylpyrimidin-2-amine was prepared according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.22 (1 H, s), 4.01 (1 H, d, J = 10.07 Hz), 3.73 (1 H, d, J = 10.32 Hz), 3.40 ( 1 H, d), 3.25 (1 H, d), 2.91-3.12 (4 H, m), 2.41 (3 H, s), 2.09-2.28 (5 H, m), 1.62-1.97 (3 H, m ). MS (LC / MS) RT = 0.47; [M + H] ⁺ = 288.31.

Example 41
(R) -N- (6-Phenylpyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(０.３２ｇ, ２.５mmol)、フェニルボロン酸(０.３８ｇ, ３.１３mmol)、飽和水性炭酸ナトリウム(０.８０ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５mmol)の混合物を、ジメトキシエタン(１５ml)／エタノール(２ml)／水(２ml)の混合物に懸濁した。混合物を１２５℃で２０分間マイクロ波で加熱し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(１０〜６０％ 酢酸エチル／ヘキサン)、６−フェニルピリミジン−４−アミンを灰白色の固体として得た(１６７mg, ０.９８mmol, 収率３９％)。
MS (LC/MS) R.T. = 0.99; [M+H]⁺ = 172.23。 Step A: 6-Phenylpyrimidin-4-amine

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 (triphenyl chloride) A mixture of phosphine) palladium (II) (0.035 g, 0.05 mmol) was suspended in a mixture of dimethoxyethane (15 ml) / ethanol (2 ml) / water (2 ml). The mixture was heated in the microwave at 125 ° C. for 20 minutes and concentrated in vacuo. The residue was purified by column chromatography (10-60% ethyl acetate / hexane) to give 6-phenylpyrimidin-4-amine as an off-white solid (167 mg, 0.98 mmol, 39% yield).
MS (LC / MS) RT = 0.99; [M + H] ⁺ = 172.23.

(Ｒ)−Ｎ−(６−フェニルピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−フェニルピリミジン−４−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 9.54 (1 H, d, J=1.01 Hz), 8.75 - 8.86 (2 H, m), 8.18 - 8.28 (3 H, m), 8.00 (1 H, br. s.), 4.70 (1 H, d, J=10.32 Hz), 4.43 (1 H, d, J=10.58 Hz), 3.72 - 3.88 (2 H, m), 3.41 - 3.63 (4 H, m), 2.63 - 2.87 (2 H, m), 2.18 - 2.48 (3 H, m). MS (LC/MS) R.T. = 1.36; [M+H]⁺ = 336.24。 Step B: (R) -N- (6-Phenylpyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-phenylpyrimidin-4-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 6-phenyl Prepared from pyrimidine-4-amine according to the general procedure of Example 23 Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 9.54 (1 H, d, J = 1.01 Hz), 8.75-8.86 (2 H, m), 8.18-8.28 (3 H, m), 8.00 (1 H, br. S.), 4.70 (1 H, d, J = 10.32 Hz), 4.43 (1 H, d, J = 10.58 Hz), 3.72-3.88 (2 H, m), 3.41-3.63 (4 H , m), 2.63-2.87 (2 H, m), 2.18-2.48 (3 H, m). MS (LC / MS) RT = 1.36; [M + H] ⁺ = 336.24.

(Ｒ)−Ｎ−(６−(４−メトキシフェニル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−クロロピリミジン−４−アミンから、実施例４１の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.72 (1 H, d, J=1.26 Hz), 7.90 - 8.00 (2 H, m), 7.16 (1 H, br. s.), 6.96 - 7.05 (2 H, m), 4.04 (1 H, d, J=10.07 Hz), 3.84 (3 H, s), 3.73 (1 H, d, J=10.07 Hz), 3.22 (1 H, d), 3.09 (1 H, d), 2.73 - 2.98 (4 H, m), 2.02 - 2.21 (2 H, m), 1.51 - 1.85 (3 H, m). MS (LC/MS) R.T. = 1.44; [M+H]⁺ = 366.28。 Example 42
(R) -N- (6- (4-Methoxyphenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (4-Methoxyphenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Was prepared from 6-chloropyrimidin-4-amine according to the general procedure of Example 41, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.72 (1 H, d, J = 1.26 Hz), 7.90-8.00 (2 H, m), 7.16 (1 H, br.s.), 6.96- 7.05 (2 H, m), 4.04 (1 H, d, J = 10.07 Hz), 3.84 (3 H, s), 3.73 (1 H, d, J = 10.07 Hz), 3.22 (1 H, d), 3.09 (1 H, d), 2.73-2.98 (4 H, m), 2.02-2.21 (2 H, m), 1.51-1.85 (3 H, m). MS (LC / MS) RT = 1.44; (M + H] ⁺ = 366.28.

(Ｒ)−Ｎ−(６−(６−メトキシピリジン−３−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−クロロピリミジン−４−アミンから、実施例４１の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.79 (2 H, dd, J=19.01, 1.89 Hz), 8.27 (1 H, dd, J=8.56, 2.52 Hz), 7.18 (1 H, br. s.), 6.89 (1 H, d, J=8.81 Hz), 4.05 (1 H, d, J=10.32 Hz), 3.96 (3 H, s), 3.74 (1 H, d, J=10.32 Hz), 3.23 (1 H, d), 3.10 (1 H, d), 2.73 - 2.99 (4 H, m), 2.02 - 2.21 (2 H, m), 1.53 - 1.84 (3 H, m). MS (LC/MS) R.T. = 1.34; [M+H]⁺ = 367.25。 Example 43
(R) -N- (6- (6-Methoxypyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (6-Methoxypyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-Amine was prepared from 6-chloropyrimidin-4-amine according to the general procedure of Example 41, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.79 (2 H, dd, J = 19.01, 1.89 Hz), 8.27 (1 H, dd, J = 8.56, 2.52 Hz), 7.18 (1 H, br s.), 6.89 (1 H, d, J = 8.81 Hz), 4.05 (1 H, d, J = 10.32 Hz), 3.96 (3 H, s), 3.74 (1 H, d, J = 10.32 Hz) ), 3.23 (1 H, d), 3.10 (1 H, d), 2.73-2.99 (4 H, m), 2.02-2.21 (2 H, m), 1.53-1.84 (3 H, m). LC / MS) RT = 1.34; [M + H] ⁺ = 367.25.

(Ｒ)−Ｎ−(６−(ナフタレン−２−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−クロロピリミジン−４−アミンから、実施例４１の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.83 (1 H, s), 8.56 (1 H, s), 7.83 - 8.13 (4 H, m), 7.49 - 7.58 (2 H, m), 7.37 (1 H, br. s.), 4.06 (1 H, d, J=10.32 Hz), 3.76 (1 H, d, J=10.32 Hz), 3.23 (1 H, s), 3.12 (1 H, d), 2.75 - 3.00 (4 H, m), 2.02 - 2.24 (2 H, m), 1.56 - 1.84 (3 H, m). MS (LC/MS) R.T. = 1.93; [M+H]⁺ = 386.31。 Example 44
(R) -N- (6- (Naphthalen-2-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (6- (Naphthalen-2-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- The amine was prepared from 6-chloropyrimidin-4-amine according to the general procedure of Example 41, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.83 (1 H, s), 8.56 (1 H, s), 7.83-8.13 (4 H, m), 7.49-7.58 (2 H, m), 7.37 (1 H, br. S.), 4.06 (1 H, d, J = 10.32 Hz), 3.76 (1 H, d, J = 10.32 Hz), 3.23 (1 H, s), 3.12 (1 H, d), 2.75-3.00 (4 H, m), 2.02-2.24 (2 H, m), 1.56-1.84 (3 H, m). MS (LC / MS) RT = 1.93; [M + H] ⁺ = 386.31.

  The compounds in Table 2 were synthesized according to the method of Example 1 using the appropriate commercially available isothiocyanate or amine. The amide-containing intermediate was obtained by the procedure described in Example 3.

Table 2

  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 dihydrochloride.

Table 3

  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 dihydrochloride.

Table 3

Example 218
(R) -N- (6- (methoxymethyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

メタノール(３０ml)中の４−メトキシ−３−オキソブタン酸メチル(３.５４ml, ２６.５mmol)の溶液に、ホルムアミジン アセテート(３.０７ｇ, ２９.２mmol)およびナトリウム メトキシド(１３ml, ５８.４mmol)を加えた。混合物を１８時間還流し、環境温度まで冷却し、濃縮した。残渣を水に溶かし、１Ｎ ＨＣｌでｐＨを７に調節した。水性混合物をクロロホルムで抽出した。合わせた有機層を硫酸マグネシウムで乾燥させ、濾過し、濃縮し、６−(メトキシメチル)ピリミジン−４−オールを得た(１.３８ｇ, ９.８５mmol, 収率３７.１％)。
MS (LC/MS) R.T. = 0.19; [M+H]⁺ = 141.20。 Step A: 6- (Methoxymethyl) pyrimidin-4-ol

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). Was added. The mixture was refluxed for 18 hours, cooled to ambient temperature and concentrated. The residue was dissolved in water and the pH was 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 give 6- (methoxymethyl) pyrimidin-4-ol (1.38 g, 9.85 mmol, 37.1% yield).
MS (LC / MS) RT = 0.19; [M + H] ⁺ = 141.20.

６−(メトキシメチル)ピリミジン−４−オール(１.３８ｇ, ９.８５mmol)をジクロロメタン(１４ml)に溶かし、オキシ塩化リン(９ml, ９７mmol)を環境温度で加えた。混合物を環境温度で１８時間撹拌し、濃縮した。残渣を氷水に溶かし、１Ｎ 水酸化ナトリウムでｐＨを７に調節した。混合物をクロロホルムで抽出し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(５〜１０％ 酢酸エチル／クロロホルム)、４−クロロ−６−(メトキシメチル)ピリミジンを薄黄色の油状物として得た(１.２ｇ, ７.５７mmol, 収率７７％)。これを静置すると固化した。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.88 (1 H, d, J=1.01 Hz), 7.52 (1 H, d, J=1.01 Hz), 4.53 (2 H, s), 3.50 (3 H, s). MS (LC/MS) R.T. = 0.98; [M+H]⁺ = 159.10。 Step B: 4-Chloro-6- (methoxymethyl) pyrimidine

6- (Methoxymethyl) pyrimidin-4-ol (1.38 g, 9.85 mmol) was dissolved in dichloromethane (14 ml) and phosphorus oxychloride (9 ml, 97 mmol) was added at ambient temperature. The mixture was stirred at ambient temperature for 18 hours and concentrated. The residue was dissolved in ice water and the pH was adjusted to 7 with 1N sodium hydroxide. The mixture was extracted with chloroform, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatography (5-10% ethyl acetate / chloroform) to give 4-chloro-6- (methoxymethyl) pyrimidine as a pale yellow oil (1.2 g, 7.57 mmol, yield). 77%). This solidified upon standing.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.88 (1 H, d, J = 1.01 Hz), 7.52 (1 H, d, J = 1.01 Hz), 4.53 (2 H, s), 3.50 (3 H , s). MS (LC / MS) RT = 0.98; [M + H] ⁺ = 159.10.

４−クロロ−６−(メトキシメチル)ピリミジン(１.２ｇ, ７.５７mmol)および水酸化アンモニウム(２０ml)の混合物を、密封管中、３時間加熱した。混合物を、環境温度まで冷却し、濃縮した。残渣をエーテルで磨砕し、６−(メトキシメチル)ピリミジン−４−アミンを薄黄色の固体として得た(０.５０ｇ, ３.５９mmol, 収率４８％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.47 (1 H, s), 6.55 (1 H, s), 5.12 (2 H, br. s.), 4.38 (2 H, s), 3.45 (3 H, s). MS (LC/MS) R.T. = 0.42; [M+H]⁺ = 140.20。 Step C: 6- (Methoxymethyl) pyrimidin-4-amine

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 give 6- (methoxymethyl) pyrimidin-4-amine as a pale yellow solid (0.50 g, 3.59 mmol, 48% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.47 (1 H, s), 6.55 (1 H, s), 5.12 (2 H, br.s.), 4.38 (2 H, s), 3.45 (3 H, s). MS (LC / MS) RT = 0.42; [M + H] ⁺ = 140.20.

１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.８４ｇ, ３.５９mmol)の明橙色の溶液に、ジクロロメタン中、環境温度で、６−(メトキシメチル)ピリミジン−４−アミン(０.５ｇ, ３.５９mmol)を加えた。橙色の溶液を環境温度で１８時間撹拌した。溶液をカラムクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル／ヘキサン)、４−イソチオシアナト−６−(メトキシメチル)ピリミジンを黄色の固体として得た(０.３２ｇ, １.７７mmol, 収率４９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.91 (1 H, d, J=1.26 Hz), 7.19 (1 H, d, J=1.01 Hz), 4.52 (2 H, s), 3.49 (3 H, s). MS (LC/MS) R.T. = 1.39; [M+H]⁺ = 182.10。 Step D: 4-isothiocyanato-6- (methoxymethyl) pyrimidine

To a light orange solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.84 g, 3.59 mmol) in dichloromethane at ambient temperature, 6- (methoxymethyl) pyrimidin-4-amine. (0.5 g, 3.59 mmol) was added. The orange solution was stirred at ambient temperature for 18 hours. The solution was purified by column chromatography (0-40% ethyl acetate / hexane) to give 4-isothiocyanato-6- (methoxymethyl) pyrimidine as a yellow solid (0.32 g, 1.77 mmol, 49% yield). ).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.91 (1 H, d, J = 1.26 Hz), 7.19 (1 H, d, J = 1.01 Hz), 4.52 (2 H, s), 3.49 (3 H , s). MS (LC / MS) RT = 1.39; [M + H] ⁺ = 182.10.

ジメチルホルムアミド中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.３９ｇ, １.７１mmol)に、炭酸セシウム(１.３９ｇ, ４.２８mmol)および４−イソチオシアナト−６−(メトキシメチル)ピリミジン(０.３１ｇ, １.７１mmol)を加えた。懸濁液を環境温度で１５分間撹拌した。反応混合物に、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.８０ml, ５.１３mmol)を加え、混合物を一夜撹拌し、濃縮した。残渣をカラムクロマトグラフィーによって精製し(酢酸エチル中５〜２５％ ９：１ メタノール／水酸化アンモニウム)、(Ｒ)−Ｎ−(６−(メトキシメチル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.１８ｇ, ０.５８mmol, 収率３４％)。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.65 (1 H, d, J=1.01 Hz), 6.92 (1 H, br. s.), 4.40 (3 H, s), 4.03 (1 H, d, J=10.32 Hz), 3.72 (1 H, d, J=10.32 Hz), 3.44 (2 H, s), 3.18 - 3.26 (1 H, m), 3.06 - 3.13 (1 H, m), 2.69 - 2.96 (4 H, m), 1.94 - 2.19 (2 H, m), 1.45 - 1.86 (3 H, m). MS (LC/MS) R.T. = 0.76; [M+H]⁺ = 304.30。 Step E: (R) -N- (6- (methoxymethyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.39 g, 1.71 mmol) in dimethylformamide was added to cesium carbonate (1.39 g, 4.28 mmol) and 4-isothiocyanato-6. -(Methoxymethyl) pyrimidine (0.31 g, 1.71 mmol) was added. The suspension was stirred at ambient temperature for 15 minutes. To the reaction mixture was added N, N′-diisopropylcarbodiimide (0.80 ml, 5.13 mmol) and the mixture was stirred overnight and concentrated. The residue was purified by column chromatography (5-25% 9: 1 methanol / ammonium hydroxide in ethyl acetate) and (R) -N- (6- (methoxymethyl) pyrimidin-4-yl) -4H-1 ′. -Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.18 g, 0.58 mmol, 34% yield).
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.65 (1 H, d, J = 1.01 Hz), 6.92 (1 H, br. S.), 4.40 (3 H, s), 4.03 (1 H , d, J = 10.32 Hz), 3.72 (1 H, d, J = 10.32 Hz), 3.44 (2 H, s), 3.18-3.26 (1 H, m), 3.06-3.13 (1 H, m), 2.69-2.96 (4 H, m), 1.94-2.19 (2 H, m), 1.45-1.86 (3 H, m). MS (LC / MS) RT = 0.76; [M + H] ⁺ = 304.30.

Example 219
(R) -N- (5- (cyclopentyloxy) pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

Ｎ,Ｎ−ジメチルホルムアミド中の、２−クロロピリミジン−５−オール(１ｇ, ７.６６mmol)、クロロシクロペンタン(２.３９ml, ２２.９８mmol)および炭酸カリウム(３.１８ｇ, ２２.９８mmol)の混合物を、６５℃で１６時間環境温度で加熱した。水を加え、混合物を酢酸エチルで抽出した。有機層を、水で、そして塩水で洗浄し、硫酸ナトリウムで乾燥させ、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(０〜２５％ 酢酸エチル／ヘキサン)、２−クロロ−５−(シクロペンチルオキシ)ピリミジンを白色の固体として得た(８３１mg, ４.１８mmol, 収率５４.６％)。
MS (LC/MS) R.T. = 2.32; [M+H]⁺ = 199.23。 Step A: 2-Chloro-5- (cyclopentyloxy) pyrimidine

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. The mixture was heated at 65 ° C. for 16 hours 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 / hexane) to give 2-chloro-5- (cyclopentyloxy) pyrimidine as a white solid (831 mg, 4.18 mmol, 54.6% yield). ).
MS (LC / MS) RT = 2.32; [M + H] ⁺ = 199.23.

５−(シクロペンチルオキシ)ピリミジン−２−アミンを、２−クロロ−５−(シクロペンチルオキシ)ピリミジンから、実施例２１８の工程Ｃの一般的な手順に従って製造した。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.99 (2 H, s), 6.18 (2 H, s), 4.54 - 4.75 (1 H, m), 1.30 - 1.91 (8 H, m). MS (LC/MS) R.T. = 1.47; [M+H]⁺ = 180.24。 Step B: 5- (Cyclopentyloxy) pyrimidin-2-amine

5- (Cyclopentyloxy) pyrimidin-2-amine was prepared from 2-chloro-5- (cyclopentyloxy) pyrimidine according to the general procedure of Example 218, Step C.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.99 (2 H, s), 6.18 (2 H, s), 4.54-4.75 (1 H, m), 1.30-1.91 (8 H, m). MS (LC / MS) RT = 1.47; [M + H] ⁺ = 180.24.

(Ｒ)−Ｎ−(５−(シクロペンチルオキシ)ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、５−(シクロペンチルオキシ)ピリミジン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.24 (2 H, s), 4.76 - 4.86 (1 H, m), 3.98 (1 H, d, J=10.07 Hz), 3.69 (1 H, d, J=10.07 Hz), 3.33 (1 H, d), 3.20 (1 H, d), 2.77 - 3.08 (4 H, m), 2.04 - 2.26 (2 H, m), 1.49 - 2.03 (11 H, m). MS (LC/MS) R.T. = 1.56; [M+H]⁺ = 344.32。 Step C: (R) -N- (5- (cyclopentyloxy) pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (5- (cyclopentyloxy) pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared from 5- (cyclopentyloxy) pyrimidin-2-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.24 (2 H, s), 4.76-4.86 (1 H, m), 3.98 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J = 10.07 Hz), 3.33 (1 H, d), 3.20 (1 H, d), 2.77-3.08 (4 H, m), 2.04-2.26 (2 H, m), 1.49-2.03 (11 H m). MS (LC / MS) RT = 1.56; [M + H] ⁺ = 344.32.

  The compounds in Table 5 were synthesized according to the method of Example 218 using the appropriate commercially available chloride as in Step C of Example 218.

Table 4

Example 228
(R) -N- (6- (2,2,2-trifluoroethoxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane ] -2-Amine

テトラヒドロフラン(１２ml)中の２,２,２−トリフルオロエタノール(２.６１ｇ, ２６.１０mmol)の溶液を、テトラヒドロフラン(４８ml)中の水素化ナトリウム(１.３１ｇ, ３２.６０mmol)の懸濁液に、０℃で滴下した。混合物を０℃で３０分間撹拌し、テトラヒドロフラン(１２ml)中の４,６−ジクロロピリミジン(３.６ｇ, ２４.１６mmol)の溶液を、０℃で加えた。反応混合物を環境温度で３時間撹拌し、飽和水性塩化アンモニウムに注ぎ、酢酸エチルで抽出した。酢酸エチル抽出物を水で洗浄し、硫酸マグネシウムで乾燥させ、真空で濃縮した。橙色の残渣をカラムクロマトグラフィーによって精製し(１０〜４０％ 酢酸エチル／ヘキサン)、４−クロロ−６−(２,２,２−トリフルオロエトキシ)ピリミジンを薄黄色の油状物として得た(２.０ｇ, ２.４１mmol, 収率３８.９％)。
MS (LC/MS) R.T. = 2.78; [M+H]⁺ = 213.12。 Step A: 4-Chloro-6- (2,2,2-trifluoroethoxy) pyrimidine

A solution of 2,2,2-trifluoroethanol (2.61 g, 26.10 mmol) in tetrahydrofuran (12 ml) was added to a suspension of sodium hydride (1.31 g, 32.60 mmol) in tetrahydrofuran (48 ml). The solution was added dropwise at 0 ° C. The mixture was stirred at 0 ° C. for 30 minutes and a solution of 4,6-dichloropyrimidine (3.6 g, 24.16 mmol) in tetrahydrofuran (12 ml) was added at 0 ° C. The reaction mixture was stirred at ambient temperature for 3 hours, poured into saturated 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 / hexane) to give 4-chloro-6- (2,2,2-trifluoroethoxy) pyrimidine as a pale yellow oil (2 0.0 g, 2.41 mmol, yield 38.9%).
MS (LC / MS) RT = 2.78; [M + H] ⁺ = 213.12.

６−(２,２,２−トリフルオロエトキシ)ピリミジン−４−アミンを、４−クロロ−６−(２,２,２−トリフルオロエトキシ)ピリミジンから、実施例２１８の工程Ｃの一般的な手順に従って製造した。
MS (LC/MS) R.T. = 1.16; [M+H]⁺ = 194.07。 Step B: 6- (2,2,2-trifluoroethoxy) pyrimidin-4-amine

6- (2,2,2-trifluoroethoxy) pyrimidin-4-amine was obtained from 4-chloro-6- (2,2,2-trifluoroethoxy) pyrimidine by the general procedure of Example 218, Step C. Manufactured according to the procedure.
MS (LC / MS) RT = 1.16; [M + H] ⁺ = 194.07.

(Ｒ)−Ｎ−(６−(２,２,２−トリフルオロエトキシ)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−(２,２,２−トリフルオロエトキシ)ピリミジン−４−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.30 (1 H, br. s.), 8.39 (1 H, s), 6.38 (1 H, br. s.), 4.59 - 4.84 (2 H, m), 3.94 (1 H, d, J=9.32 Hz), 3.59 (1 H, d, J=9.57 Hz), 3.33 (1 H, d, J=16.62 Hz), 2.61 - 3.01 (5 H, m), 1.97 - 2.25 (2 H, m), 1.37 - 1.81 (3 H, m). MS (LC/MS) R.T. = 1.42; [M+H]⁺ = 358.33。 Step C: (R) -N- (6- (2,2,2-trifluoroethoxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2. 2] Octane] -2-amine

(R) -N- (6- (2,2,2-trifluoroethoxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane ] -2-amine was prepared from 6- (2,2,2-trifluoroethoxy) pyrimidin-4-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.30 (1 H, br. S.), 8.39 (1 H, s), 6.38 (1 H, br. S.), 4.59-4.84 (2 H, m ), 3.94 (1 H, d, J = 9.32 Hz), 3.59 (1 H, d, J = 9.57 Hz), 3.33 (1 H, d, J = 16.62 Hz), 2.61-3.01 (5 H, m) , 1.97-2.25 (2 H, m), 1.37-1.81 (3 H, m). MS (LC / MS) RT = 1.42; [M + H] ⁺ = 358.33.

Example 229
(R) -N- (5-Bromo-4-isopropylpyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

４−イソプロピルピリミジン−２−アミンを、２−クロロ−４−イソプロピルピリミジンから、実施例２１８の工程Ｃの一般的な手順に従って製造した。
¹H NMR (400 MHz, DMSO-d₆) d ppm 8.10 (1 H, d, J=5.04 Hz), 6.45 (3 H, d, J=5.04 Hz), 2.59 - 2.80 (1 H, m), 1.15 (6 H, d, J=7.05 Hz). MS (LC/MS) R.T. = 0.76; [M+H]⁺ = 138.12。 Step A: 4-Isopropylpyrimidin-2-amine

4-Isopropylpyrimidin-2-amine was prepared from 2-chloro-4-isopropylpyrimidine according to the general procedure of Example 218, Step C.
¹ H NMR (400 MHz, DMSO-d ₆ ) d ppm 8.10 (1 H, d, J = 5.04 Hz), 6.45 (3 H, d, J = 5.04 Hz), 2.59-2.80 (1 H, m), 1.15 (6 H, d, J = 7.05 Hz). MS (LC / MS) RT = 0.76; [M + H] ⁺ = 138.12.

Ｎ−ブロモスクシンイミド(０.５ｇ, ２.８mmol)を、クロロホルム中の４−イソプロピルピリミジン−２−アミン(０.３９ｇ, ２.８１mmol)の溶液に加えた。得られた黄色の溶液を環境温度で１時間撹拌し、真空で濃縮した。残渣をカラムクロマトグラフィーによって精製し(クロロホルム中３〜１０％ ９：１ メタノール：水酸化アンモニウム)、５−ブロモ−４−イソプロピルピリミジン−２−アミンを薄黄色の固体として得た(０.６９ｇ, ３.１８mmol, １１３％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.21 (1 H, s), 6.75 (2 H, s), 3.10 - 3.23 (1 H, m), 1.14 (6 H, d, J=6.80 Hz). MS (LC/MS) R.T. = 2.58; [M]⁺ = 216.09。 Step B: 5-Bromo-4-isopropylpyrimidin-2-amine

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 resulting yellow solution was stirred at ambient temperature for 1 hour and concentrated in vacuo. The residue was purified by column chromatography (3-10% 9: 1 methanol: ammonium hydroxide in chloroform) to give 5-bromo-4-isopropylpyrimidin-2-amine as a pale yellow solid (0.69 g, 3.18 mmol, 113%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.21 (1 H, s), 6.75 (2 H, s), 3.10-3.23 (1 H, m), 1.14 (6 H, d, J = 6.80 Hz). MS (LC / MS) RT = 2.58; [M] ⁺ = 216.09.

(Ｒ)−Ｎ−(５−ブロモ−４−イソプロピルピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、５−ブロモ−４−イソプロピルピリミジン−２−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.49 (1 H, s), 4.02 (1 H, d, J=10.07 Hz), 3.72 (1 H, d, J=10.07 Hz), 3.34 - 3.44 (1 H, m), 3.23 (1 H, s), 3.06 - 3.15 (1 H, m), 2.95 (2 H, t, J=7.55 Hz), 2.75 - 2.89 (2 H, m), 2.00 - 2.21 (2 H, m), 1.52 - 1.83 (3 H, m), 1.24 (6 H, d, J=6.80 Hz). MS (LC/MS) R.T. = 1.84; [M+H]⁺ = 382.24。 Step C: (R) -N- (5-Bromo-4-isopropylpyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (5-Bromo-4-isopropylpyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine , 5-bromo-4-isopropylpyrimidin-2-amine, prepared according to the general procedure of Example 23 Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.49 (1 H, s), 4.02 (1 H, d, J = 10.07 Hz), 3.72 (1 H, d, J = 10.07 Hz), 3.34- 3.44 (1 H, m), 3.23 (1 H, s), 3.06-3.15 (1 H, m), 2.95 (2 H, t, J = 7.55 Hz), 2.75-2.89 (2 H, m), 2.00 -2.21 (2 H, m), 1.52-1.83 (3 H, m), 1.24 (6 H, d, J = 6.80 Hz). MS (LC / MS) RT = 1.84; [M + H] ⁺ = 382.24 .

(Ｒ)−Ｎ−(５−ブロモ−４−(ピリジン−３−イル)ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−(ピリジン−３−イル)ピリミジン−２−アミンから、実施例２２９の工程Ｂ〜Ｃの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.92 (1 H, d, J=1.51 Hz), 8.73 (1 H, s), 8.64 (1 H, dd, J=4.91, 1.64 Hz), 8.23 (1 H, dt, J=8.06, 1.89 Hz), 7.56 (1 H, dd, J=7.93, 4.91 Hz), 4.00 (1 H, d, J=10.07 Hz), 3.69 (1 H, d, J=10.07 Hz), 3.22 (1 H, d), 3.07 (1 H, d), 2.60 - 2.99 (4 H, m), 2.00 - 2.21 (2 H, m), 1.50 - 1.83 (3 H, m). MS (LC/MS) R.T. = 0.76; [M+H]⁺ = 416.30。 Example 230
(R) -N- (5-Bromo-4- (pyridin-3-yl) pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane ] -2-Amine

(R) -N- (5-Bromo-4- (pyridin-3-yl) pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane ] -2-amine was prepared from 4- (pyridin-3-yl) pyrimidin-2-amine according to the general procedure of Examples 229, steps B-C.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.92 (1 H, d, J = 1.51 Hz), 8.73 (1 H, s), 8.64 (1 H, dd, J = 4.91, 1.64 Hz), 8.23 (1 H, dt, J = 8.06, 1.89 Hz), 7.56 (1 H, dd, J = 7.93, 4.91 Hz), 4.00 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J = 10.07 Hz), 3.22 (1 H, d), 3.07 (1 H, d), 2.60-2.99 (4 H, m), 2.00-2.21 (2 H, m), 1.50-1.83 (3 H, m ). MS (LC / MS) RT = 0.76; [M + H] ⁺ = 416.30.

Example 231
(R) -N- (6- (cyclopentyloxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

テトラヒドロフラン(１２ml)中のシクロペンタノール(２.２５ｇ, ２６.１mmol)の溶液を、テトラヒドロフラン(４８ml)中の水素化ナトリウム(１.３１ｇ, ３２.６mmol)の懸濁液に、０℃で滴下した。混合物を０℃で３０分間撹拌し、テトラヒドロフラン(１２ml)中の４,６−ジクロロピリミジン(３.６ｇ, ２４.１６mmol)の溶液を、０℃で加えた。反応混合物を環境温度で３時間撹拌し、飽和水性塩化アンモニウムに注ぎ、酢酸エチルで抽出した。有機層を水で洗浄し、硫酸マグネシウムで乾燥させ、真空で濃縮した。有機残渣をカラムクロマトグラフィーによって精製し(１０〜４０％ 酢酸エチル／ヘキサン)、４−クロロ−６−(シクロペンチルオキシ)ピリミジンを薄黄色の油状物として得た。この物質を次の反応に直接用いた。 Step A: 4-Chloro-6- (cyclopentyloxy) pyrimidine

A solution of cyclopentanol (2.25 g, 26.1 mmol) in tetrahydrofuran (12 ml) was added dropwise at 0 ° C. to a suspension of sodium hydride (1.31 g, 32.6 mmol) in tetrahydrofuran (48 ml). did. The mixture was stirred at 0 ° C. for 30 minutes and a solution of 4,6-dichloropyrimidine (3.6 g, 24.16 mmol) in tetrahydrofuran (12 ml) was added at 0 ° C. The reaction mixture was stirred at ambient temperature for 3 hours, poured into saturated aqueous ammonium chloride and extracted with ethyl acetate. The organic layer was washed with water, dried over magnesium sulfate and concentrated in vacuo. The organic residue was purified by column chromatography (10-40% ethyl acetate / hexane) to give 4-chloro-6- (cyclopentyloxy) pyrimidine as a pale yellow oil. This material was used directly in the next reaction.

６−(シクロペンチルオキシ)ピリミジン−４−アミンを、４−クロロ−６−(シクロペンチルオキシ)ピリミジンから、実施例２１８の工程Ｃの一般的な手順に従って製造した。
MS (LC/MS) R.T. = 1.64; [M+H]⁺ = 180.22。 Step B: 6- (Cyclopentyloxy) pyrimidin-4-amine

6- (Cyclopentyloxy) pyrimidin-4-amine was prepared from 4-chloro-6- (cyclopentyloxy) pyrimidine according to the general procedure of Example 218, Step C.
MS (LC / MS) RT = 1.64; [M + H] ⁺ = 180.22.

(Ｒ)−Ｎ−(６−(シクロペンチルオキシ)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、６−(シクロペンチルオキシ)ピリミジン−４−アミンから、実施例２３の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.37 (1 H, s), 6.16 (1 H, br. s.), 5.27 (1 H, br. s.), 3.98 (1 H, d, J=10.32 Hz), 3.67 (1 H, d, J=10.32 Hz), 3.15 - 3.24 (1 H, m), 3.02 - 3.12 (1 H, m), 2.71 - 2.97 (4 H, m), 2.01 - 2.14 (2 H, m), 1.87 - 2.00 (3 H, m), 1.49 - 1.84 (8 H, m). MS (LC/MS) R.T. = 1.96; [M+H]⁺ = 344.34。 Step C: (R) -N- (6- (cyclopentyloxy) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (6- (cyclopentyloxy) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared from 6- (cyclopentyloxy) pyrimidin-4-amine according to the general procedure of Example 23, Steps AB.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.37 (1 H, s), 6.16 (1 H, br. S.), 5.27 (1 H, br. S.), 3.98 (1 H, d , J = 10.32 Hz), 3.67 (1 H, d, J = 10.32 Hz), 3.15-3.24 (1 H, m), 3.02-3.12 (1 H, m), 2.71-2.97 (4 H, m), 2.01-2.14 (2 H, m), 1.87-2.00 (3 H, m), 1.49-1.84 (8 H, m). MS (LC / MS) RT = 1.96; [M + H] ⁺ = 344.34.

(Ｒ)−Ｎ−(６−イソプロポキシピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４,６−ジクロロピリミジンから、実施例２３１の工程Ａ〜Ｃの一般的な手順に従って製造した。
¹H NMR (400 MHz, MeOD-d₄) δ ppm 8.37 (1 H, s), 6.13 (1 H, br. s.), 5.09 - 5.30 (1 H, m), 3.98 (1 H, d, J=10.32 Hz), 3.67 (1 H, d, J=10.32 Hz), 3.13 - 3.24 (1 H, m), 3.01 - 3.09 (1 H, m), 2.68 - 2.98 (4 H, m), 1.98 - 2.17 (2 H, m), 1.49 - 1.83 (3 H, m), 1.30 (6 H, d, J=6.04 Hz). MS (LC/MS) R.T. = 1.36; [M+H]⁺ = 318.24。 Example 232
(R) -N- (6-Isopropoxypyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Isopropoxypyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 4, Prepared from 6-dichloropyrimidine according to the general procedure of Examples 231, Steps AC.
¹ H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.37 (1 H, s), 6.13 (1 H, br.s.), 5.09-5.30 (1 H, m), 3.98 (1 H, d, J = 10.32 Hz), 3.67 (1 H, d, J = 10.32 Hz), 3.13-3.24 (1 H, m), 3.01-3.09 (1 H, m), 2.68-2.98 (4 H, m), 1.98 -2.17 (2 H, m), 1.49-1.83 (3 H, m), 1.30 (6 H, d, J = 6.04 Hz). MS (LC / MS) RT = 1.36; [M + H] ⁺ = 318.24 .

(Ｒ)−Ｎ−(６−(２,２−ジフルオロエトキシ)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４,６−ジクロロピリミジンから、実施例２３１の工程Ａ〜Ｃの一般的な手順に従って製造した。
融点 83〜8℃. 1H NMR (400 MHz, MeOD-d₄) δ ppm 8.42 (1 H, s), 5.93 - 6.37 (2 H, m), 4.52 (2 H, td, J=13.98, 3.78 Hz), 3.99 (1 H, d, J=10.32 Hz), 3.68 (1 H, d, J=10.32 Hz), 3.19 (1 H, d), 3.07 (1 H, d), 2.67 - 2.97 (4 H, m), 1.99 - 2.19 (2 H, m), 1.51 - 1.82 (3 H, m). MS (LC/MS) R.T. = 0.99; [M+H]⁺ = 340.26。 Example 233
(R) -N- (6- (2,2-difluoroethoxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (6- (2,2-difluoroethoxy) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 The amine was prepared from 4,6-dichloropyrimidine according to the general procedure of steps 231 C of Example 231.
Melting point 83-8 ° C. 1H NMR (400 MHz, MeOD-d ₄ ) δ ppm 8.42 (1 H, s), 5.93-6.37 (2 H, m), 4.52 (2 H, td, J = 13.98, 3.78 Hz ), 3.99 (1 H, d, J = 10.32 Hz), 3.68 (1 H, d, J = 10.32 Hz), 3.19 (1 H, d), 3.07 (1 H, d), 2.67-2.97 (4 H , m), 1.99-2.19 (2 H, m), 1.51-1.82 (3 H, m). MS (LC / MS) RT = 0.99; [M + H] ⁺ = 340.26.

(Ｒ)−Ｎ−(ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、(Ｒ)−Ｎ−(６−ブロモピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(実施例１５５より)から、実施例１９の工程Ｃの一般的な手順に従って製造した。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 8.26 (d, J=4.88 Hz, 1 H), 7.58 - 7.74 (m, 1 H), 6.85 - 7.02 (m, 2 H), 4.00 (d, J=10.07 Hz, 1 H), 3.70 (d, J=10.07 Hz, 1 H), 3.26 - 3.35 (m, 1 H), 3.14 - 3.21 (m, 1 H), 3.02 (d, J=8.24 Hz, 2 H), 2.84 - 2.97 (m, 2 H), 2.11 - 2.25 (m, 2 H), 1.58 - 1.92 (m, 3 H). MS (LC/MS) R.T. = 0.30; [M+H]⁺ = 259.16。 Example 234
(R) -N- (Pyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to (R) —N— From (6-bromopyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine (from Example 155) to Example 19 Prepared according to the general procedure of Step C.
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 8.26 (d, J = 4.88 Hz, 1 H), 7.58-7.74 (m, 1 H), 6.85-7.02 (m, 2 H), 4.00 (d , J = 10.07 Hz, 1 H), 3.70 (d, J = 10.07 Hz, 1 H), 3.26-3.35 (m, 1 H), 3.14-3.21 (m, 1 H), 3.02 (d, J = 8.24 Hz, 2 H), 2.84-2.97 (m, 2 H), 2.11-2.25 (m, 2 H), 1.58-1.92 (m, 3 H). MS (LC / MS) RT = 0.30; (M + H ] ⁺ = 259.16.

(Ｒ)−Ｎ−(ピリジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、(Ｒ)−Ｎ−(２−ブロモピリジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(実施例１５４より)から、実施例１９の工程Ｃの手順に従って製造した。
¹H NMR (500 MHz, MeOD-d₄) δ ppm 8.31 (d, J=6.41 Hz, 2 H), 7.39 (d, J=3.97 Hz, 2 H), 4.01 (d, J=12.21 Hz, 1 H), 3.70 (d, J=11.90 Hz, 1 H), 3.37 (s, 1 H), 3.29 (s, 1 H), 3.18 (d, J=1.83 Hz, 1 H), 3.15 (d, J=2.14 Hz, 1 H), 2.86 - 3.09 (m, 3 H), 2.02 - 2.20 (m, 1 H), 1.59 - 1.88 (m, 3 H). MS (LC/MS) R.T. = 0.22; [M+H]⁺ = 259.16。 Example 235
(R) -N- (pyridin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (pyridin-4-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to (R) —N— From (2-bromopyridin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine (from Example 154) to Example 19 Prepared according to the procedure of Step C.
¹ H NMR (500 MHz, MeOD-d ₄ ) δ ppm 8.31 (d, J = 6.41 Hz, 2 H), 7.39 (d, J = 3.97 Hz, 2 H), 4.01 (d, J = 12.21 Hz, 1 H), 3.70 (d, J = 11.90 Hz, 1 H), 3.37 (s, 1 H), 3.29 (s, 1 H), 3.18 (d, J = 1.83 Hz, 1 H), 3.15 (d, J = 2.14 Hz, 1 H), 2.86-3.09 (m, 3 H), 2.02-2.20 (m, 1 H), 1.59-1.88 (m, 3 H). MS (LC / MS) RT = 0.22; (M + H] ⁺ = 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] Octane] -2-amine

チオシアン酸カリウム(１２.４２ｇ, １２８mmol)を酢酸(４５.０ml)に懸濁し、０℃に冷却した。国際公開公報第2006/044707号に従って製造した６−(ベンジルオキシ)ピリジン−３−アミン(３.２ｇ, １５.９８mmol)を加えた。酢酸(１５ml)中の臭素(２.５５ml, ４９.５mmol)を３０分かけて滴下し、その間に反応混合物は非常に濃厚になった。それをゆっくりと室温まで温め、一夜撹拌した。水(２０ml)を加え、反応混合物を９０℃に加熱し、熱濾過した。濾液を残して、フィルターケーキを反応フラスコに戻し、これにさらに４０mlのＨＯＡｃを加えた。混合物を再度９０℃に加熱し、熱濾過した。合わせた濾液を氷浴で冷却し、ｐＨ＞８になるまでＮＨ_４ＯＨを滴下した。黄色の沈殿物が形成し、これを濾過によって集めた。固体を真空で１時間乾燥させ、５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−アミンを得た(１.９５ｇ, ７.５８mmol, 収率４７.４％)。これをさらに精製することなく用いた。
¹H NMR (400 MHz, CDCl₃) δ ppm 7.70 (d, J=8.78 Hz, 1 H) 7.48 (d, J=7.28 Hz, 2 H) 7.39 (t, J=7.28 Hz, 2 H) 7.30 - 7.36 (m, 1 H) 6.78 (d, J=8.78 Hz, 1 H) 5.39 (s, 2 H) 5.14 (br. s., 2 H)。 Step A: 5- (Benzyloxy) thiazolo [5,4-b] pyridin-2-amine

Potassium thiocyanate (12.42 g, 128 mmol) was suspended in acetic acid (45.0 ml) and cooled to 0 ° C. 6- (Benzyloxy) pyridin-3-amine (3.2 g, 15.98 mmol) prepared according to WO 2006/044707 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 slowly warmed to room temperature and stirred overnight. Water (20 ml) was added and the reaction mixture was heated to 90 ° C. and filtered hot. The filter cake was returned to the reaction flask, leaving the filtrate, to which an additional 40 ml of HOAc was added. The mixture was heated again to 90 ° C. and filtered hot. The combined filtrates were cooled in an ice bath and NH ₄ OH was added dropwise until pH> 8. A yellow precipitate formed and was collected by filtration. The solid was dried in vacuo for 1 hour to give 5- (benzyloxy) thiazolo [5,4-b] pyridin-2-amine (1.95 g, 7.58 mmol, 47.4% yield). This was used without further purification.
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.70 (d, J = 8.78 Hz, 1 H) 7.48 (d, J = 7.28 Hz, 2 H) 7.39 (t, J = 7.28 Hz, 2 H) 7.30- 7.36 (m, 1 H) 6.78 (d, J = 8.78 Hz, 1 H) 5.39 (s, 2 H) 5.14 (br. S., 2 H).

ＤＭＦ(３.１ml)中の５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−アミン(８００mg, ３.１１mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(０.３ml, ６.２２mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で、二硫化炭素(０.４７ml, ７.７７mmol)を加え、混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(０.３ml, ６.２２mmol)を加え、混合物を再度１０分間撹拌した。最終的にヨードメタン(０.４７ml, ７.４６mmol)を滴下した。混合物を１時間撹拌し、この時点で、それを水に注ぎ、ＥｔＯＡｃ(３×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。粗製の混合物をシリカゲルのクロマトグラフィーによって精製し(２〜２０％ ＥｔＯＡｃ／ＣＨＣｌ_３)、５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルを黄色の結晶性固体として得た(１.０２ｇ, 収率９１％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.02 (d, J=8.78 Hz, 1 H) 7.51 (d, J=7.28 Hz, 2 H) 7.38 - 7.45 (m, 2 H) 7.32 - 7.38 (m, 1 H) 6.89 (d, J=8.53 Hz, 1 H) 5.46 (s, 2 H) 2.65 (s, 6 H)。 Step B: Dimethyl 5- (benzyloxy) thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate

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 20.0 M sodium hydroxide (0.3 ml). , 6.22 mmol). The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (0.47 ml, 7.77 mmol) was added and the mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (0.3 ml, 6.22 mmol) was added and the mixture was stirred again for 10 minutes. Finally iodomethane (0.47 ml, 7.46 mmol) was added dropwise. The mixture was stirred for 1 hour, at which point it was poured into water and extracted with EtOAc (3x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by chromatography on silica gel (2-20% EtOAc / CHCl ₃ ), and dimethyl 5- (benzyloxy) thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate was obtained as yellow crystals. (1.02 g, 91% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.02 (d, J = 8.78 Hz, 1 H) 7.51 (d, J = 7.28 Hz, 2 H) 7.38-7.45 (m, 2 H) 7.32-7.38 (m , 1 H) 6.89 (d, J = 8.53 Hz, 1 H) 5.46 (s, 2 H) 2.65 (s, 6 H).

ＤＭＦ(７ml)中の、５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(５００mg, １.３８mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(３１７mg, １.３８mmol)および炭酸セシウム(１.０ｇ, ３.０７mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。混合物をシリカゲルのクロマトグラフィーによって精製し(２〜２０％ [９：１ メタノール：水酸化アンモニウム]−クロロホルム)、(Ｒ)−Ｎ−(５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(３９０mg, 収率６７％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.18 (br. s., 1 H) 7.76 (d, J=8.85 Hz, 1 H) 7.50 (d, J=7.32 Hz, 2 H) 7.41 (t, J=7.32 Hz, 2 H) 7.32 - 7.37 (m, 1 H) 5.42 (s, 2 H) 4.02 (d, J=9.46 Hz, 1 H) 3.68 (d, J=9.46 Hz, 1 H) 3.37 - 3.44 (m, 1 H) 2.75 - 3.06 (m, 5 H) 2.13 - 2.25 (m, 2 H) 1.73 - 1.82 (m, 1 H) 1.49 - 1.70 (m, 3 H). MS (LC/MS) R.T. = 1.69; [M+H]⁺ = 421.98。 Step C: (R) -N- (5- (Benzyloxy) thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2 .2] Octane] -2-amine

Dimethyl 5- (benzyloxy) thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate (500 mg, 1.38 mmol), (S) -3- (aminomethyl) quinuclidine in DMF (7 ml). A mixture of -3-ol dihydrochloride (317 mg, 1.38 mmol) and cesium carbonate (1.0 g, 3.07 mmol) was heated at 100 ° C. for 2 hours. The reaction mixture was cooled to ambient temperature, poured into water and extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The mixture was purified by chromatography on silica gel (2-20% [9: 1 methanol: ammonium hydroxide] -chloroform) and (R) -N- (5- (benzyloxy) thiazolo [5,4-b] pyridine. -2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (390 mg, 67% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.18 (br.s., 1 H) 7.76 (d, J = 8.85 Hz, 1 H) 7.50 (d, J = 7.32 Hz, 2 H) 7.41 (t, J = 7.32 Hz, 2 H) 7.32-7.37 (m, 1 H) 5.42 (s, 2 H) 4.02 (d, J = 9.46 Hz, 1 H) 3.68 (d, J = 9.46 Hz, 1 H) 3.37- 3.44 (m, 1 H) 2.75-3.06 (m, 5 H) 2.13-2.25 (m, 2 H) 1.73-1.82 (m, 1 H) 1.49-1.70 (m, 3 H). MS (LC / MS) RT = 1.69; [M + H] ⁺ = 421.98.

(Ｒ)−Ｎ−(５−(ベンジルオキシ)チアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(３９０mg, ０.９２５mmol)をＴＦＡに溶解し、環境温度で４時間反応させた。この時点で、ＬＣＭＳおよびＴＬＣにより、出発物質がほとんど消費されたことが示された。ＴＦＡを真空で除去し、粗製の混合物を分取ＨＰＬＣによって精製した。合わせた生成物のフラクションを真空で濃縮し、エーテルで磨砕し、(Ｒ)−２−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)チアゾロ[５,４−ｂ]ピリジン−５(４Ｈ)−オンのＴＦＡ塩を得た(１６４mg, ０.３６８mmol, 収率３９.８％)。
融点 245℃(分解). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.99 (br. s., 1 H) 9.05 (br. s., 1 H) 7.81 (d, J=8.53 Hz, 1 H) 6.65 (d, J=8.78 Hz, 1 H) 3.96 (d, J=10.29 Hz, 1 H) 3.82 (d, J=10.54 Hz, 1 H) 3.63 - 3.78 (m, 2 H) 3.36 - 3.47 (m, 1 H) 3.16 - 3.34 (m, 3 H) 2.43 (br. s., 1 H) 2.16 (br. s., 1 H) 1.76 - 2.07 (m, 3 H). MS (LC/MS) R.T. = 0.50; [M+H]⁺ = 332.15。 Example 237
(R) -2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) thiazolo [5,4-b] pyridine-5 (4H) -ON

(R) -N- (5- (Benzyloxy) thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine (390 mg, 0.925 mmol) was dissolved in TFA and allowed to react for 4 hours at ambient temperature. At this point, LCMS and TLC showed that most of the starting material was consumed. TFA was removed in vacuo and the crude mixture was purified by preparative HPLC. The combined product fractions were concentrated in vacuo, triturated with ether and (R) -2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- The TFA salt of 2-ylamino) thiazolo [5,4-b] pyridin-5 (4H) -one was obtained (164 mg, 0.368 mmol, yield 39.8%).
Mp 245 ° C. ^{(decomposition). 1 H NMR (400 MHz} , DMSO-d 6) δ ppm 9.99 (br. S., 1 H) 9.05 (br. S., 1 H) 7.81 (d, J = 8.53 Hz, 1 H) 6.65 (d, J = 8.78 Hz, 1 H) 3.96 (d, J = 10.29 Hz, 1 H) 3.82 (d, J = 10.54 Hz, 1 H) 3.63-3.78 (m, 2 H) 3.36- 3.47 (m, 1 H) 3.16-3.34 (m, 3 H) 2.43 (br. S., 1 H) 2.16 (br. S., 1 H) 1.76-2.07 (m, 3 H). MS (LC / MS) RT = 0.50; [M + H] ⁺ = 332.15.

Example 238
(R) -N- (6- (3-methoxyphenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(０.３２４ｇ, ２.５mmol)、３−メトキシフェニルボロン酸(０.４７５ｇ, ３.１３mmol)、Ｎａ_２ＣＯ_３(０.７９５ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５０mmol)の混合物を、ＤＭＥ／ＥｔＯＨ／水(１５：２：３ml)に懸濁し、マイクロ波合成装置中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(１０〜６０％ 酢酸エチル−ヘキサン)、６−(３−メトキシフェニル)ピリミジン−４−アミンを灰白色の固体として得た(０.３５ｇ, １.７４mmol, 収率７０％)。
LCMS R.T. = 1.28; [M+H]⁺ = 201.98。 Step A: 6- (3-Methoxyphenyl) pyrimidin-4-amine

6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol), 3-methoxyphenylboronic acid (0.475 g, 3.13 mmol), Na ₂ CO ₃ (0.795 g, 7.50 mmol) and bis chloride A mixture of (triphenylphosphine) palladium (II) (0.035 g, 0.050 mmol) is suspended in DME / EtOH / water (15: 2: 3 ml) and is kept in a microwave synthesizer at 125 ° C. for 20 minutes. Heated and concentrated. The residue was purified by chromatography on silica gel (10-60% ethyl acetate-hexane) to give 6- (3-methoxyphenyl) pyrimidin-4-amine as an off-white solid (0.35 g, 1.74 mmol, yield). (Rate 70%).
LCMS RT = 1.28; [M + H] ⁺ = 201.98.

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.９７０ｇ, ４.１７mmol)の溶液に、室温で、６−(３−メトキシフェニル)ピリミジン−４−アミン(０.７ｇ, ３.４８mmol)を加えた。反応物を室温で１８時間撹拌した。ＬＣ／ＭＳにより、主要なピークとして望ましい生成物のピークが示された。深橙色の溶液を濃縮し、残った残渣を濾過した。濾液をシリカゲルのクロマトグラフィーによって精製し(０〜１０％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−(３−メトキシフェニル)ピリミジンを黄色の油状物として得た(０.３９ｇ, ４.３１mmol, 収率４６％)。
LCMS R.T. = 2.91; [M+H]⁺ = 244.03。 Step B: 4-isothiocyanato-6- (3-methoxyphenyl) pyrimidine

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.970 g, 4.17 mmol) in dichloromethane at room temperature, 6- (3-methoxyphenyl) pyrimidin-4-amine (0 0.7 g, 3.48 mmol) was added. The reaction was stirred at room temperature for 18 hours. LC / MS showed the desired product peak as the main peak. The deep orange solution was concentrated and the remaining residue was filtered. The filtrate was purified by chromatography on silica gel (0-10% ethyl acetate-hexane) to give 4-isothiocyanato-6- (3-methoxyphenyl) pyrimidine as a yellow oil (0.39 g, 4.31 mmol, Yield 46%).
LCMS RT = 2.91; [M + H] ⁺ = 244.03.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.３６３ｇ, １.５８３mmol)に、Ｃｓ_２ＣＯ_３(１.２８９ｇ, ３.９６mmol)および４−イソチオシアナト−６−(３−メトキシフェニル)ピリミジンを加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.７４０ml, ４.７５mmol)を加え、混合物を室温で１８時間撹拌し続けた。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(６−メトキシピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを薄黄色の溶液として得た(０.２９４ｇ, ０.７８８mmol, 収率５０％)。
融点 80〜5℃. ¹H NMR (400 MHz, MeOD) δ ppm 8.77 (1 H, s), 7.47 - 7.58 (2 H, m), 7.39 (1 H, t), 7.20 (1 H, br. s.), 7.04 (1 H, dd), 4.05 (1 H, d), 3.85 (3 H, s), 3.74 (1 H, d), 3.23 (1 H, d), 3.10 (1 H, d), 2.71 - 3.00 (4 H, m), 2.03 - 2.22 (2 H, m), 1.53 - 1.85 (3 H, m). MS (LC/MS) R.T. = 1.58; [M+H]⁺ = 366.15。 Step C: (R) -N- (6- (3-methoxyphenyl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane]- 2-Amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.363 g, 1.583 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (1.289 g, 3 .96 mmol) and 4-isothiocyanato-6- (3-methoxyphenyl) pyrimidine were added. The suspension was stirred at room temperature for 30 minutes. N, N′-diisopropylcarbodiimide (0.740 ml, 4.75 mmol) was added and the mixture continued to stir at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (6-methoxypyrimidin-4-yl) -4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a pale yellow solution (0.294 g, 0.788 mmol, 50% yield).
Melting point 80-5 ° C. ¹ H NMR (400 MHz, MeOD) δ ppm 8.77 (1 H, s), 7.47-7.58 (2 H, m), 7.39 (1 H, t), 7.20 (1 H, br. s.), 7.04 (1 H, dd), 4.05 (1 H, d), 3.85 (3 H, s), 3.74 (1 H, d), 3.23 (1 H, d), 3.10 (1 H, d ), 2.71-3.00 (4 H, m), 2.03-2.22 (2 H, m), 1.53-1.85 (3 H, m). MS (LC / MS) RT = 1.58; [M + H] ⁺ = 366.15 .

Example 239
(R) -N- (isoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.８０５ｇ, ３.４７mmol)の溶液に、室温で、イソキノリン−３−アミン(０.５ｇ, ３.４７mmol)を加えた。反応物を室温で１８時間撹拌した。ＬＣ／ＭＳにより、主要なピークとして望ましい生成物のピークが示された。深橙色の溶液を濃縮し、濾過した。濾液をシリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−(３−メトキシフェニル)ピリミジンを白色の固体として得た(０.５５ｇ, ２.９６mmol, 収率８５％)。
LCMS R.T. = 2.47; [M+H]⁺ = 187.23。 Step A: 3-isothiocyanatoisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.805 g, 3.47 mmol) in dichloromethane at room temperature is isoquinolin-3-amine (0.5 g, 3.47 mmol). added. The reaction was stirred at room temperature for 18 hours. LC / MS showed the desired product peak as the main peak. The deep orange solution was concentrated and filtered. The filtrate was purified by chromatography on silica gel (0-40% ethyl acetate-hexane) to give 4-isothiocyanato-6- (3-methoxyphenyl) pyrimidine as a white solid (0.55 g, 2.96 mmol, yield). (Rate 85%).
LCMS RT = 2.47; [M + H] ⁺ = 187.23.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２ｇ, ０.８７３mmol)に、Ｃｓ_２ＣＯ_３(０.７１１ｇ, ２.１８２mmol)および３−イソチオシアナトイソキノリン(０.１６３ｇ, ０.８７３mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.４０８ml, ２.６２mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ [９：１ メタノール：水酸化アンモニウム]−酢酸エチル)、(Ｒ)−Ｎ−(イソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを灰白色の固体として得た(０.１６ｇ, ０.５０８mmol, 収率５８％)。
融点 196〜200℃. ¹H NMR (400 MHz, MeOD) δ ppm 9.00 (1 H, s), 7.92 (1 H, d), 7.71 (1 H, d), 7.59 (1 H, t), 7.20 - 7.45 (2 H, m), 3.96 (1 H, d), 3.65 (1 H, d), 3.22 (1 H, d), 3.08 (1 H, d), 2.66 - 3.00 (4 H, m), 2.05 - 2.23 (2 H, m), 1.50 - 1.86 (3 H, m). R.T. = 1.37; [M+H]⁺ = 309.31。 Step B: (R) -N- (isoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.2 g, 0.873 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.711 g, 2 .182 mmol) and 3-isothiocyanatoisoquinoline (0.163 g, 0.873 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-diisopropylcarbodiimide (0.408 ml, 2.62 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% [9: 1 methanol: ammonium hydroxide] -ethyl acetate), (R) -N- (isoquinolin-3-yl) -4H-1 ′. -Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as an off-white solid (0.16 g, 0.508 mmol, 58% yield).
Melting point 196-200 ° C. ¹ H NMR (400 MHz, MeOD) δ ppm 9.00 (1 H, s), 7.92 (1 H, d), 7.71 (1 H, d), 7.59 (1 H, t), 7.20 -7.45 (2 H, m), 3.96 (1 H, d), 3.65 (1 H, d), 3.22 (1 H, d), 3.08 (1 H, d), 2.66-3.00 (4 H, m) , 2.05-2.23 (2 H, m), 1.50-1.86 (3 H, m). RT = 1.37; [M + H] ⁺ = 309.31.

Example 240
(R) -N- (6-phenoxypyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(３.００ｇ, ２３.１４mmol)を、フェノール(１１.２９ｇ, １２０mmol)中のナトリウム(０.１９７ｇ, ８.５７mmol)の溶液に、５５℃で加えた。混合物を１４０℃で２時間加熱し、室温で２０時間維持した。反応混合物を、氷／水上で、混合物の温度を２０℃未満に保ちながら、３２％ 水性ＮａＯＨに注いだ。混合物をクロロホルムで抽出し、有機抽出物を塩化カルシウムで乾燥させ、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中２〜２０％ 酢酸エチル)、６−フェノキシピリミジン−４−アミンを白色の固体として得た(０.６ｇ, ３.２１mmol, 収率７５％)。
LCMS R.T. = 1.37; [M+H]⁺ = 197.95。 Step A: 6-Phenoxypyrimidin-4-amine

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 ° C. The mixture was heated at 140 ° C. for 2 hours and maintained at room temperature for 20 hours. The reaction mixture was poured onto 32% aqueous NaOH on ice / water, keeping the temperature of the mixture below 20 ° C. The mixture was extracted with chloroform and the organic extract was dried over calcium chloride and concentrated. The residue was purified by chromatography on silica gel (2-20% ethyl acetate in hexanes) to give 6-phenoxypyrimidin-4-amine as a white solid (0.6 g, 3.21 mmol, 75% yield).
LCMS RT = 1.37; [M + H] ⁺ = 197.95.

ＤＣＭ中の６−フェノキシピリミジン−４−アミン(０.２８８ｇ, １.５３８mmol)および１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.３５７ｇ, １.５３８mmol)の混合物を、室温で１８時間撹拌した。薄橙色の混合物をシリカゲルのクロマトグラフィーによって精製し(５〜３５％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−フェノキシピリミジンを黄色の油状物として得た(０.５５ｇ, ２.９６mmol, 収率８５％)。
LCMS R.T. = 2.78; [M+H]⁺ = 229.94。 Step B: 4-isothiocyanato-6-phenoxypyrimidine

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 added. Stir at room temperature for 18 hours. The pale orange mixture was purified by chromatography on silica gel (5-35% ethyl acetate-hexane) to give 4-isothiocyanato-6-phenoxypyrimidine as a yellow oil (0.55 g, 2.96 mmol, yield). 85%).
LCMS RT = 2.78; [M + H] ⁺ = 229.94.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１７０ｇ, ０.７４２mmol)に、Ｃｓ_２ＣＯ_３(０.６０４ｇ, １.８５４mmol)および４−イソチオシアナト−６−フェノキシピリミジン(０.１７ｇ, ０.７４２mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.３４７ml, ２.２２５mmol)を加え、混合物を室温で１８時間撹拌し続けた。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(６−フェノキシピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを薄黄色の固体として得た(０.２１ｇ, ０.７２mmol, 収率４８.２％)。
¹H NMR (500 MHz, MeOD) δ ppm 8.43 (1 H, s), 7.47 (2 H, t), 7.30 (1 H, t), 7.16 (2 H, d), 6.21 (1 H, br. s.), 4.03 (1 H, d), 3.72 (1 H, d), 3.22 (1 H, d), 3.11 (1 H, d), 2.73 - 2.99 (4 H, m), 2.00 - 2.18 (2 H, m), 1.54 - 1.88 (3 H, m). LCMS R.T. = 1.46; [M+H]⁺ = 352.19。 Step C: (R) -N- (6-phenoxypyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.170 g, 0.742 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.604 g, 1 .854 mmol) and 4-isothiocyanato-6-phenoxypyrimidine (0.17 g, 0.742 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.347 ml, 2.225 mmol) was added and the mixture was kept stirring at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (6-phenoxypyrimidin-4-yl) -4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a pale yellow solid (0.21 g, 0.72 mmol, 48.2% yield).
¹ H NMR (500 MHz, MeOD) δ ppm 8.43 (1 H, s), 7.47 (2 H, t), 7.30 (1 H, t), 7.16 (2 H, d), 6.21 (1 H, br. s.), 4.03 (1 H, d), 3.72 (1 H, d), 3.22 (1 H, d), 3.11 (1 H, d), 2.73-2.99 (4 H, m), 2.00-2.18 ( 2 H, m), 1.54-1.88 (3 H, m). LCMS RT = 1.46; [M + H] ⁺ = 352.19.

Example 241
(R) -N- (7-methoxyquinoxalin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

J. Chem. Soc. Perk Trans. 1, 2001, 978-984に従って製造した２−クロロ−７−メトキシキノキサリン(０.５１ｇ, ２.６２mmol)、および、(２,４−ジメトキシフェニル)メタンアミン(１.１８１ml, ７.８６mmol)を、ＤＭＳＯ(２.５ml)中で、１５０℃で３０分間マイクロ波照射した。これを１５０mlのＥｔＯＡｃで希釈し、１００mlの塩水で３回抽出した。粗生成物を、９０ｇのシリカゲルのカートリッジで、ヘキサン中２０〜８０％ ＥｔＯＡｃ、５０分、４０ml/分でフラッシュクロマトグラフィーによって精製し、Ｎ−(２,４−ジメトキシベンジル)−７−メトキシキノキサリン−２−アミンを得た(７９５mg, ２.４４３mmol, 収率９３％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.00 (1 H, s), 7.70 (1 H, d, J=8.81 Hz), 7.29 (1 H, d, J=8.31 Hz), 7.05 (1 H, d, J=2.77 Hz), 6.97 (1 H, dd, J=9.06, 2.77 Hz), 6.47 (1 H, d, J=2.27 Hz), 6.43 (1 H, dd, J=8.18, 2.39 Hz), 5.22 (1 H, t, J=5.29 Hz), 4.63 (2 H, d, J=5.54 Hz), 3.91 (3 H, s), 3.83 (3 H, s), 3.78 (3 H, s). LCMS: RT = 1.91分, MH⁺ = 326.15。 Step A: N- (2,4-Dimethoxybenzyl) -7-methoxyquinoxalin-2-amine

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) was microwaved in DMSO (2.5 ml) at 150 ° C. for 30 minutes. This was diluted with 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 20-80% EtOAc in hexanes, 50 min, 40 ml / min to give N- (2,4-dimethoxybenzyl) -7-methoxyquinoxaline- 2-Amine was obtained (795 mg, 2.443 mmol, 93% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.00 (1 H, s), 7.70 (1 H, d, J = 8.81 Hz), 7.29 (1 H, d, J = 8.31 Hz), 7.05 (1 H , d, J = 2.77 Hz), 6.97 (1 H, dd, J = 9.06, 2.77 Hz), 6.47 (1 H, d, J = 2.27 Hz), 6.43 (1 H, dd, J = 8.18, 2.39 Hz ), 5.22 (1 H, t, J = 5.29 Hz), 4.63 (2 H, d, J = 5.54 Hz), 3.91 (3 H, s), 3.83 (3 H, s), 3.78 (3 H, s LCMS: RT = 1.91 min, MH ⁺ = 326.15.

Ｎ−(２,４−ジメトキシベンジル)−７−メトキシキノキサリン−２−アミン(０.７９ｇ, ２.４２８mmol)を、ＴＦＡ(１０ml, １３０mmol)／ＣＨ_２Ｃｌ_２(１０ml)中で、室温で３０分間撹拌した。溶媒をロータリーエバポレーターで除去した。飽和水性ＮａＨＣＯ_３(２００ml)を赤色の残渣に加えると、黄色の固体が沈殿した。混合物を大量のＤＣＭで抽出した。有機層を濃縮し、真空で乾燥させ、７−メトキシキノキサリン−２−アミン ２,２,２−トリフルオロアセテートを得た(０.７０ｇ, ２.４mmol, 収率９９％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.10 (1 H, s), 7.63 (1 H, d, J=9.07 Hz), 6.95 (1 H, dd, J=9.06, 2.77 Hz), 6.89 (1 H, d, J=2.77 Hz), 6.85 (2 H, br. s.), 3.84 (3 H, s). LCMS: RT = 1.04 分, MH⁺ = 176.14。 Step B: 7-Methoxyquinoxalin-2-amine 2,2,2-trifluoroacetate

N- (2,4-dimethoxybenzyl) -7-methoxyquinoxalin-2-amine (0.79 g, 2.428 mmol) was added in TFA (10 ml, 130 mmol) / CH ₂ Cl ₂ (10 ml) at room temperature for 30 minutes. Stir for minutes. The solvent was removed on a rotary evaporator. Saturated aqueous NaHCO ₃ (200 ml) was added to the red residue and a yellow solid precipitated. The mixture was extracted with a large amount of DCM. The organic layer was concentrated and dried in vacuo to give 7-methoxyquinoxalin-2-amine 2,2,2-trifluoroacetate (0.70 g, 2.4 mmol, 99% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.10 (1 H, s), 7.63 (1 H, d, J = 9.07 Hz), 6.95 (1 H, dd, J = 9.06, 2.77 Hz), 6.89 (1 H, d, J = 2.77 Hz), 6.85 (2 H, br. S.), 3.84 (3 H, s). LCMS: RT = 1.04 min, MH ⁺ = 176.14.

７−メトキシキノキサリン−２−アミン ２,２,２−トリフルオロアセテート(５７８mg, ２mmol)、トリエチルアミン(３３５μｌ, ２.４００mmol)および１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(５５７mg, ２.４００mmol)の混合物を、５mlのＤＣＭ中で２４時間撹拌した。反応物を、１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分で直接溶出し、２−イソチオシアナト−７−メトキシキノキサリンを得た(８４mg, ０.３８７mmol, 収率１９％)。
LCMS: RT = 2.49分, MH⁺ = 218.06。 Step C: 2-isothiocyanato-7-methoxyquinoxaline

7-methoxyquinoxalin-2-amine 2,2,2-trifluoroacetate (578 mg, 2 mmol), triethylamine (335 μl, 2.400 mmol) and 1,1′-thiocarbonyldipyridin-2 (1H) -one (557 mg) , 2.400 mmol) was stirred in 5 ml DCM for 24 hours. The reaction was eluted directly on a 120 g silica gel cartridge with 0-25% EtOAc in hexanes, 25 min, 35 ml / min to give 2-isothiocyanato-7-methoxyquinoxaline (84 mg, 0.387 mmol, yield). 19%).
LCMS: RT = 2.49 min, MH ⁺ = 218.06.

(Ｒ)−Ｎ−(７−メトキシキノキサリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ｂの方法によって合成した。フラッシュクロマトグラフィーを、１２０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜４％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分で行い、２４mgを得た(収率１７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.73 (1 H, br. s.), 8.45 (1 H, s), 7.80 (1 H, d, J=9.07 Hz), 7.12 (1 H, dd, J=9.06, 2.77 Hz), 7.03 (1 H, d, J=2.52 Hz), 4.02 (1 H, d, J=9.32 Hz), 3.90 (3 H, s), 3.67 (1 H, d, J=9.32 Hz), 3.36 (1 H, dd, J=14.86, 1.51 Hz), 2.69 - 3.06 (5 H, m), 2.10 - 2.26 (2 H, m), 1.66 - 1.80 (1 H, m), 1.43 - 1.63 (2 H, m). LCMS: RT = 0.835分, MH^- = 338.2, MH⁺ = 340.1。 Step D: (R) -N- (7-Methoxyquinoxalin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (7-methoxyquinoxalin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 23. It was synthesized by the method of Step B. Flash chromatography was performed on a 120 g silica gel cartridge with 1-4% [9: 1 MeOH / NH ₄ OH] in CHCl _{3 for} 50 minutes to give 24 mg (17% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.73 (1 H, br.s.), 8.45 (1 H, s), 7.80 (1 H, d, J = 9.07 Hz), 7.12 (1 H, dd , J = 9.06, 2.77 Hz), 7.03 (1 H, d, J = 2.52 Hz), 4.02 (1 H, d, J = 9.32 Hz), 3.90 (3 H, s), 3.67 (1 H, d, J = 9.32 Hz), 3.36 (1 H, dd, J = 14.86, 1.51 Hz), 2.69-3.06 (5 H, m), 2.10-2.26 (2 H, m), 1.66-1.80 (1 H, m) , 1.43 - 1.63 (2 H, 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] octane] -2-amine

J. Chem. Soc. 1948 ,1310-1313に従って製造した２−クロロ−６−メチルキノキサリン(０.５１ｇ, ２.８６mmol)、および、(２,４−ジメトキシフェニル)メタンアミン(１.２９ml, ８.５７mmol)を、ＤＭＳＯ(２.５ml)中、１５０℃で３０分間マイクロ波照射した。これを１５０mlのＥｔＯＡｃで希釈し、１００mlの塩水で３回抽出した。粗生成物を、フラッシュクロマトグラフィーによって、９０ｇのシリカゲルのカートリッジで、ヘキサン中２０〜６０％ ＥｔＯＡｃ、５０分、４０ml/分で精製し、Ｎ−(２,４−ジメトキシベンジル)−６−メチルキノキサリン−２−アミンを得た(８４８mg, ２.７４mmol, 収率９６％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.12 (1 H, s), 7.60 (1 H, s), 7.59 (1 H, d, J=5.79 Hz), 7.38 (1 H, dd, J=8.56, 1.76 Hz), 7.30 (1 H, d, J=8.31 Hz), 6.46 (1 H, d, J=2.27 Hz), 6.42 (1 H, dd, J=8.18, 2.39 Hz), 5.20 (1 H, t, J=5.41 Hz), 4.62 (2 H, d, J=5.54 Hz), 3.83 (3 H, s), 3.77 (3 H, s), 2.46 (3 H, s). LCMS: RT = 1.93分, MH⁺ = 310.20。 Step A: N- (2,4-dimethoxybenzyl) -6-methylquinoxalin-2-amine

2-Chloro-6-methylquinoxaline (0.51 g, 2.86 mmol) and (2,4-dimethoxyphenyl) methanamine (1.29 ml, 8.10) prepared according to J. Chem. Soc. 1948, 1310-1313. 57 mmol) was microwaved in DMSO (2.5 ml) at 150 ° C. for 30 minutes. This was diluted with 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 20-60% EtOAc in hexanes, 50 min, 40 ml / min, and N- (2,4-dimethoxybenzyl) -6-methylquinoxaline. 2-Amine was obtained (848 mg, 2.74 mmol, 96% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.12 (1 H, s), 7.60 (1 H, s), 7.59 (1 H, d, J = 5.79 Hz), 7.38 (1 H, dd, J = 8.56, 1.76 Hz), 7.30 (1 H, d, J = 8.31 Hz), 6.46 (1 H, d, J = 2.27 Hz), 6.42 (1 H, dd, J = 8.18, 2.39 Hz), 5.20 (1 H, t, J = 5.41 Hz), 4.62 (2 H, d, J = 5.54 Hz), 3.83 (3 H, s), 3.77 (3 H, s), 2.46 (3 H, s). LCMS: RT = 1.93 min, MH ⁺ = 310.20.

Ｎ−(２,４−ジメトキシベンジル)−６−メチルキノキサリン−２−アミン (０.８４ｇ, ２.７２mmol)を、ＴＦＡ(１０ml, １３０mmol)／ＣＨ_２Ｃｌ_２(１０ml)中で、室温で３０分間撹拌した。溶媒をロータリーエバポレーターで除去した。飽和水性Ｎａ_２ＣＯ_３(２００ml)を赤色の残渣に加えると、黄褐色の固体が沈殿した。混合物を大量のＤＣＭで抽出した。有機層を硫酸ナトリウムで乾燥させ、濃縮し、真空で乾燥させ、６−メチルキノキサリン−２−アミン ２,２,２−トリフルオロアセテートを得た(６４０mg, ２.３４３mmol, 収率８６％)。
1H NMR (400 MHz, DMSO-d₆) δ ppm 8.24 (1 H, s), 7.55 (1 H, s), 7.34 - 7.45 (2 H, m), 6.82 (2 H, s), 2.41 (3 H, s). LCMS: RT = 1.07 分, MH⁺ = 160.12。 Step B: 6-Methylquinoxalin-2-amine 2,2,2-trifluoroacetate

N- (2,4-dimethoxybenzyl) -6-methylquinoxalin-2-amine (0.84 g, 2.72 mmol) was added in TFA (10 ml, 130 mmol) / CH ₂ Cl ₂ (10 ml) at room temperature for 30 minutes. Stir for minutes. The solvent was removed on a rotary evaporator. Saturated aqueous Na ₂ CO ₃ (200 ml) was added to the red residue and a tan solid precipitated. The mixture was extracted with a large amount of DCM. The organic layer was dried over sodium sulfate, concentrated and dried in vacuo to give 6-methylquinoxalin-2-amine 2,2,2-trifluoroacetate (640 mg, 2.343 mmol, 86% yield).
1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.24 (1 H, s), 7.55 (1 H, s), 7.34-7.45 (2 H, m), 6.82 (2 H, s), 2.41 (3 H, s). LCMS: RT = 1.07 min, MH ⁺ = 160.12.

６−メチルキノキサリン−２−アミン ２,２,２−トリフルオロアセテート(５４６mg, ２mmol)、トリエチルアミン(２４３mg, ２.４００mmol)および１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(５５７mg, ２.４０mmol)の混合物を、５mlのＤＣＭ中で４時間撹拌した。反応物を、直接、１２０ｇのシリカゲルのカートリッジから、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分で溶出し、２−イソチオシアナト−６−メチルキノキサリンを得た(１５３mg, ０.７６０mmol, 収率３８％)。
LCMS: RT = 2.59分, MH⁺ = 202.04。 Step C: 2-isothiocyanato-6-methylquinoxaline

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 hours. The reaction was eluted directly from a 120 g silica gel cartridge with 0-25% EtOAc in hexanes, 25 min, 35 ml / min to give 2-isothiocyanato-6-methylquinoxaline (153 mg, 0.760 mmol, yield). (Rate 38%).
LCMS: RT = 2.59 min, MH ⁺ = 202.04.

(Ｒ)−Ｎ−(６−メチルキノキサリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ｂの方法によって合成した。１２０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜４％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分でフラッシュクロマトグラフィーを行い、４６mgを得た(収率１９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.75 (1 H, br. s.), 8.57 (1 H, s), 7.71 (1 H, s), 7.60 (1 H, d, J=8.56 Hz), 7.42 (1 H, dd, J=8.44, 1.89 Hz), 4.01 (1 H, d, J=9.57 Hz), 3.68 (1 H, d, J=9.32 Hz), 3.38 (1 H, dd, J=14.86, 1.01 Hz), 2.73 - 3.08 (5 H, m), 2.50 (3 H, s), 2.16 - 2.26 (1 H, m), 2.14 (1 H, br. s.), 1.67 - 1.79 (1 H, m), 1.45 - 1.65 (2 H, m). LCMS: RT = 0.838分, MH^- = 322.2, MH⁺ = 324.2。 Step D: (R) -N- (6-Methylquinoxalin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Methylquinoxalin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 23. It was synthesized by the method of Step B. Flash chromatography on a 120 g silica gel cartridge with 1-4% [9: 1 MeOH / NH ₄ OH] in CHCl _{3 for} 50 minutes yielded 46 mg (19% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.75 (1 H, br. S.), 8.57 (1 H, s), 7.71 (1 H, s), 7.60 (1 H, d, J = 8.56 Hz ), 7.42 (1 H, dd, J = 8.44, 1.89 Hz), 4.01 (1 H, d, J = 9.57 Hz), 3.68 (1 H, d, J = 9.32 Hz), 3.38 (1 H, dd, J = 14.86, 1.01 Hz), 2.73-3.08 (5 H, m), 2.50 (3 H, s), 2.16-2.26 (1 H, m), 2.14 (1 H, br.s.), 1.67-1.79 . (1 H, m), 1.45 - 1.65 (2 H, m) LCMS: RT = 0.838 ^{min, MH - = 322.2, MH +} = 324.2.

Example 243
(R) -N- (7-Methylquinoxalin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

J. Chem. Soc. 1948, 1310-1313に従って製造した２−クロロ−７−メチルキノキサリン(０.５１ｇ, ２.８６mmol)、および、(２,４−ジメトキシフェニル)メタンアミン(１.２９ml, ８.５７mmol)に、ＤＭＳＯ(２.５ml)中、１５０℃で３０分間マイクロ波照射した。これを１５０mlのＥｔＯＡｃで希釈し、１００mlの塩水で３回抽出した。粗生成物を、フラッシュクロマトグラフィーによって、９０ｇのシリカゲルのカートリッジで、ヘキサン中２０〜８０％ ＥｔＯＡｃ、５０分、４０ml/分で精製し、Ｎ−(２,４−ジメトキシベンジル)−７−メチルキノキサリン−２−アミンを得た(８６０mg, ２.７８mmol, 収率９７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.08 (1 H, s), 7.70 (1 H, d, J=8.31 Hz), 7.49 (1 H, s), 7.30 (1 H, d, J=8.06 Hz), 7.17 (1 H, dd, J=8.31, 2.01 Hz), 6.47 (1 H, d, J=2.52 Hz), 6.42 (1 H, dd, J=8.31, 2.52 Hz), 5.23 (1 H, t, J=5.16 Hz), 4.63 (2 H, d, J=5.79 Hz), 3.83 (3 H, s), 3.78 (3 H, s), 2.48 (3 H, s). LCMS: RT = 1.93分, MH⁺ = 310.20。 Step A: N- (2,4-Dimethoxybenzyl) -7-methylquinoxalin-2-amine

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, 8.10). 57 mmol) was microwaved in DMSO (2.5 ml) at 150 ° C. for 30 minutes. This was diluted with 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 20-80% EtOAc in hexanes, 50 min, 40 ml / min, and N- (2,4-dimethoxybenzyl) -7-methylquinoxaline. 2-Amine was obtained (860 mg, 2.78 mmol, 97% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.08 (1 H, s), 7.70 (1 H, d, J = 8.31 Hz), 7.49 (1 H, s), 7.30 (1 H, d, J = 8.06 Hz), 7.17 (1 H, dd, J = 8.31, 2.01 Hz), 6.47 (1 H, d, J = 2.52 Hz), 6.42 (1 H, dd, J = 8.31, 2.52 Hz), 5.23 (1 H, t, J = 5.16 Hz), 4.63 (2 H, d, J = 5.79 Hz), 3.83 (3 H, s), 3.78 (3 H, s), 2.48 (3 H, s). LCMS: RT = 1.93 min, MH ⁺ = 310.20.

Ｎ−(２,４−ジメトキシベンジル)−７−メチルキノキサリン−２−アミン(０.８５ｇ, ２.７５mmol)を、ＴＦＡ(１０ml, １３０mmol)／ＣＨ_２Ｃｌ_２(１０ml)中で、室温で３０分間撹拌した。溶媒をロータリーエバポレーターで除去した。飽和水性ＮａＨＣＯ_３(２００ml)を赤色の残渣に加えると、桃色の固体が沈殿した。混合物を大量のＤＣＭで抽出した。有機層を硫酸ナトリウムで乾燥させ、濃縮し、真空下で乾燥させ、７−メチルキノキサリン−２−アミン ２,２,２−トリフルオロアセテートを得た(６４０mg, ２.３４mmol, 収率８５％)。
1H NMR (400 MHz, DMSO-d₆) δ ppm 8.20 (1 H, s), 7.64 (1 H, d, J=8.31 Hz), 7.28 (1 H, s), 7.15 (1 H, dd, J=8.31, 1.76 Hz), 6.89 (2 H, s), 2.42 (3 H, s). LCMS: RT = 1.07分, MH⁺ = 160.12。 Step B: 7-Methylquinoxalin-2-amine 2,2,2-trifluoroacetate

N- (2,4-dimethoxybenzyl) -7-methylquinoxalin-2-amine (0.85 g, 2.75 mmol) was added in TFA (10 ml, 130 mmol) / CH ₂ Cl ₂ (10 ml) at room temperature for 30 minutes. Stir for minutes. The solvent was removed on a rotary evaporator. Saturated aqueous NaHCO ₃ (200 ml) was added to the red residue and a pink solid precipitated. The mixture was extracted with a large amount of DCM. The organic layer was dried over sodium sulfate, concentrated and dried under vacuum to give 7-methylquinoxalin-2-amine 2,2,2-trifluoroacetate (640 mg, 2.34 mmol, 85% yield). .
1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.20 (1 H, s), 7.64 (1 H, d, J = 8.31 Hz), 7.28 (1 H, s), 7.15 (1 H, dd, J = 8.31, 1.76 Hz), 6.89 (2 H, s), 2.42 (3 H, s). LCMS: RT = 1.07 min, MH ⁺ = 160.12.

７−メチルキノキサリン−２−アミン ２,２,２−トリフルオロアセテート(５４６mg, ２mmol)(78263-058-01)、トリエチルアミン(２４３mg, ２.４０mmol)および１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(５５７mg, ２.４０mmol)の混合物を、５mlのＤＣＭ中で２時間撹拌した。反応物を、直接、１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分で溶出し、２−イソチオシアナト−７−メチルキノキサリンを得た(１８５mg, ０.９１９mmol, 収率４６％)。
LCMS: RT = 2.58分, MH⁺ = 202.04。 Step C: 2-isothiocyanato-6-methylquinoxaline

7-Methylquinoxalin-2-amine 2,2,2-trifluoroacetate (546 mg, 2 mmol) (78263-058-01), triethylamine (243 mg, 2.40 mmol) and 1,1′-thiocarbonyldipyridine-2 A mixture of (1H) -one (557 mg, 2.40 mmol) was stirred in 5 ml DCM for 2 hours. The reaction was directly eluted with a 120 g silica gel cartridge eluting with 0-25% EtOAc in hexanes, 25 min, 35 ml / min to give 2-isothiocyanato-7-methylquinoxaline (185 mg, 0.919 mmol, yield). Rate 46%).
LCMS: RT = 2.58 min, MH ⁺ = 202.04.

(Ｒ)−Ｎ−(７−メチルキノキサリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ｂの方法によって合成した。１２０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜３％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分でフラッシュクロマトグラフィーを行い、２２mgを得た(収率７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.80 (1 H, br. s.), 8.53 (1 H, s), 7.81 (1 H, d, J=8.31 Hz), 7.50 (1 H, s), 7.31 (1 H, dd, J=8.56, 1.76 Hz), 4.01 (1 H, d, J=9.57 Hz), 3.66 (1 H, d, J=9.32 Hz), 3.36 (1 H, d, J=14.86 Hz), 2.70 - 3.04 (5 H, m), 2.50 (3 H, s), 2.15 - 2.24 (1 H, m), 2.13 (1 H, br. s.), 1.66 - 1.79 (1 H, m), 1.44 - 1.63 (2 H, m). LCMS: RT = 8.67分, MH^- = 322.6, MH⁺ = 324.1。 Step D: (R) -N- (7-Methylquinoxalin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (7-Methylquinoxalin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 23. It was synthesized by the method of Step B. Flash chromatography on a 120 g silica gel cartridge with 1-3% [9: 1 MeOH / NH ₄ OH] in CHCl _{3 for} 50 minutes yielded 22 mg (7% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.80 (1 H, br.s.), 8.53 (1 H, s), 7.81 (1 H, d, J = 8.31 Hz), 7.50 (1 H, s ), 7.31 (1 H, dd, J = 8.56, 1.76 Hz), 4.01 (1 H, d, J = 9.57 Hz), 3.66 (1 H, d, J = 9.32 Hz), 3.36 (1 H, d, J = 14.86 Hz), 2.70-3.04 (5 H, m), 2.50 (3 H, s), 2.15-2.24 (1 H, m), 2.13 (1 H, br.s.), 1.66-1.79 (1 . H, m), 1.44 - 1.63 (2 H, 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′-bicyclo [2.2.2] octane] -2- Amine

６−クロロピリミジン−４−アミン(０.３２４ｇ, ２.５mmol)、ピリジン−３−イルボロン酸(０.３８４ｇ, ３.１３mmol)、Ｎａ_２ＣＯ_３(０.７９５ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５０mmol)の混合物を、ＤＭＥ／ＥｔＯＨ／水の混合物に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中１０〜６０％ 酢酸エチル、次に５〜２５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、６−(ピリジン−３−イル)ピリミジン−４−アミンを灰白色の固体として得た(０.１７ｇ, ０.９８７mmol, 収率４０％)。
LCMS R.T. = 0.31; [M+H]⁺ = 173.11。 Step A: 6- (Pyridin-3-yl) pyrimidin-4-amine

6-Chloro-pyrimidin-4-amine (0.324g, 2.5mmol), 3-ylboronic acid _{(0.384g, 3.13mmol), Na 2} CO 3 (0.795g, 7.50mmol) and bis A mixture of (triphenylphosphine) palladium (II) (0.035 g, 0.050 mmol) was suspended in a mixture of DME / EtOH / water. The mixture was heated in a microwave synthesizer at 125 ° C. for 20 minutes and concentrated. The residue was purified by chromatography on silica gel (10-60% ethyl acetate in hexane, then 5-25% 9: 1 methanol: ammonium hydroxide-ethyl acetate), 6- (pyridin-3-yl) pyrimidine-4 -The amine was obtained as an off-white solid (0.17 g, 0.987 mmol, 40% yield).
LCMS RT = 0.31; [M + H] ⁺ = 173.11.

ジクロロメタン／Ｎ,Ｎ−ジメチルホルムアミド中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.６８２ｇ, ２.９４mmol)の溶液に、室温で、６−(ピリジン−３−イル)ピリミジン−４−アミン(０.３３７ｇ, １.９５７mmol)を加えた。混合物を６０℃で１８時間加熱した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物を、シリカゲルのクロマトグラフィーによって精製し(１〜４０％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−メトキシピリミジンを橙色の油状物として得た(０.１２ｇ, ０.５６mmol, 収率２８.６％)。 Step B: 4-Isothiocyanato-6- (pyridin-3-yl) pyrimidine

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.682 g, 2.94 mmol) in dichloromethane / N, N-dimethylformamide at room temperature, 6- (pyridin-3-yl Pyrimidin-4-amine (0.337 g, 1.957 mmol) was added. The mixture was heated at 60 ° C. for 18 hours. LC / MS showed the desired product peak as the major peak. The deep orange mixture was purified by chromatography on silica gel (1-40% ethyl acetate-hexane) to give 4-isothiocyanato-6-methoxypyrimidine as an orange oil (0.12 g, 0.56 mmol, yield). (Rate 28.6%).

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１３ｇ, ０.５６０mmol)に、Ｃｓ_２ＣＯ_３(０.４６ｇ, １.４mmol)および４−イソチオシアナト−６−(ピリジン−３−イル)ピリミジン(０.１２ｇ, ０.５６mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.２６ml, １.７mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(０〜１０％ [９：１ メタノール：水酸化アンモニウム]−酢酸エチル)、(Ｓ)−Ｎ−(５−クロロピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを灰白色の固体として得た(０.１８２ｇ, ０.６１３mmol, 収率３５％)。
¹H NMR (400 MHz, MeOD) δ ppm 9.13 - 9.21 (1 H, m), 8.82 (1 H, d), 8.63 (1 H, dd), 8.44 (1 H, dt), 7.56 (1 H, dd), 7.31 (1 H, s), 4.06 (1 H, d), 3.76 (1 H, d), 3.20 - 3.28 (1 H, m), 3.08 - 3.16 (1 H, m), 2.72 - 3.01 (4 H, m), 2.00 - 2.24 (2 H, m), 1.52 - 1.83 (3 H, m). LCMS R.T. = 0.72; [M+H]⁺ = 337.2。 Step C: (R) -N- (6- (Pyridin-3-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.13 g, 0.560 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.46 g, 1 0.4 mmol) and 4-isothiocyanato-6- (pyridin-3-yl) pyrimidine (0.12 g, 0.56 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.26 ml, 1.7 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (0-10% [9: 1 methanol: ammonium hydroxide] -ethyl acetate), (S) -N- (5-chloropyrazin-2-yl) -4H. -1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as an off-white solid (0.182 g, 0.613 mmol, 35% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 9.13-9.21 (1 H, m), 8.82 (1 H, d), 8.63 (1 H, dd), 8.44 (1 H, dt), 7.56 (1 H, dd), 7.31 (1 H, s), 4.06 (1 H, d), 3.76 (1 H, d), 3.20-3.28 (1 H, m), 3.08-3.16 (1 H, m), 2.72-3.01 (4 H, m), 2.00-2.24 (2 H, m), 1.52-1.83 (3 H, m). LCMS RT = 0.72; [M + H] ⁺ = 337.2.

Example 245
(R) -N- (2′-methoxy-4,5′-bipyrimidin-6-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

６−クロロピリミジン−４−アミン(０.３５ｇ, ２.７０mmol)、２−メトキシピリミジン−５−イルボロン酸(０.５２０ｇ, ３.３８mmol)、Ｎａ_２ＣＯ_３(０.８５９ｇ, ８.１１mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３８ｇ, ０.０５４mmol)を、ＤＭＥ／ＥｔＯＨ／水の混合物(１５：２：３ml)に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(０〜５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、６−(ピリジン−３−イル)ピリミジン−４−アミンを灰白色の固体として得た(０.２８ｇ, １.３７８mmol, 収率５１％)。
LCMS R.T. = 0.53; [M+H]⁺ = 204.11。 Step A: 2'-methoxy-4,5'-bipyrimidin-6-amine

6-chloropyrimidin-4-amine (0.35 g, 2.70 mmol), 2-methoxypyrimidin-5-ylboronic acid (0.520 g, 3.38 mmol), Na ₂ CO 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 a microwave synthesizer at 125 ° C. for 20 minutes and concentrated. The residue was purified by chromatography on silica gel (0-5% 9: 1 methanol: ammonium hydroxide-ethyl acetate) to give 6- (pyridin-3-yl) pyrimidin-4-amine as an off-white solid (0 .28 g, 1.378 mmol, 51% yield).
LCMS RT = 0.53; [M + H] ⁺ = 204.11.

ジクロロメタン／Ｎ,Ｎ−ジメチルホルムアミド中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.８３２ｇ, ３.５８mmol)の溶液に、室温で、２'−メトキシ−４,５'−ビピリミジン−６−アミン(０.５６ｇ, ２.７６mmol)を加えた。橙色の混合物を６０℃で１８時間加熱した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物を、シリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−メトキシピリミジンを橙色の固体として得た(０.１ｇ, ０.４０８mmol, 収率１５％)。
LCMS R.T. = 2.29; [M+H]⁺ = 246.03。 Step B: 6-Isothiocyanato-2′-methoxy-4,5′-bipyrimidine

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.832 g, 3.58 mmol) in dichloromethane / N, N-dimethylformamide at room temperature, 2′-methoxy-4,5 '-Bipyrimidin-6-amine (0.56 g, 2.76 mmol) was added. The orange mixture was heated at 60 ° C. for 18 hours. LC / MS showed the desired product peak as the major peak. The deep orange mixture was purified by chromatography on silica gel (0-40% ethyl acetate-hexane) to give 4-isothiocyanato-6-methoxypyrimidine as an orange solid (0.1 g, 0.408 mmol, yield). 15%).
LCMS RT = 2.29; [M + H] ⁺ = 246.03.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.０９３ｇ, ０.４１mmol)に、Ｃｓ_２ＣＯ_３(０.３３ｇ, １mmol)および６−イソチオシアナト−２'−メトキシ−４,５'−ビピリミジン(０.１ｇ, ０.４１mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１９ml, １.２mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ [９：１ メタノール：水酸化アンモニウム]−酢酸エチル)、(Ｒ)−Ｎ−(２'−メトキシ−４,５'−ビピリミジン−６−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを灰白色の固体として得た(０.０７２ｇ, ０.１８８mmol, 収率４６％)。
¹H NMR (400 MHz, MeOD) δ ppm 9.19 (2 H, s), 8.80 (1 H, d), 7.24 (1 H, br. s.), 4.00 - 4.09 (4 H, m), 3.76 (1 H, d), 3.23 (1 H, s), 3.08 - 3.15 (1 H, m), 2.72 - 3.04 (4 H, m), 1.97 - 2.22 (2 H, m), 1.38 - 1.85 (3 H, m). R.T. = 1.22; [M+H]⁺ = 368.22。 Step C: (R) -N- (2′-methoxy-4,5′-bipyrimidin-6-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.093 g, 0.41 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.33 g, 1 mmol) and 6-isothiocyanato-2′-methoxy-4,5′-bipyrimidine (0.1 g, 0.41 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.19 ml, 1.2 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% [9: 1 methanol: ammonium hydroxide] -ethyl acetate), (R) -N- (2′-methoxy-4,5′-bipyrimidine. -6-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as an off-white solid (0.072 g, 0.188 mmol, Yield 46%).
¹ H NMR (400 MHz, MeOD) δ ppm 9.19 (2 H, s), 8.80 (1 H, d), 7.24 (1 H, br.s.), 4.00-4.09 (4 H, m), 3.76 ( 1 H, d), 3.23 (1 H, s), 3.08-3.15 (1 H, m), 2.72-3.04 (4 H, m), 1.97-2.22 (2 H, m), 1.38-1.85 (3 H m). RT = 1.22; [M + H] ⁺ = 368.22.

Example 246
(R) -N- (6- (Pyridin-4-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- Amine

６−クロロピリミジン−４−アミン(０.３２４ｇ, ２.５mmol)、ピリジン−４−イルボロン酸(０.３８４ｇ, ３.１３mmol)、Ｎａ_２ＣＯ_３(０.７９５ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５０mmol)を、ＤＭＥ／ＥｔＯＨ／水(１５：２：３ml)の混合物に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(５〜２５％ [９：１ メタノール：水酸化アンモニウム]−酢酸エチル)、６−(ピリジン−３−イル)ピリミジン−４−アミンを灰白色の固体として得た(０.１５ｇ, ０.８７１mmol, 収率３５％)。
LCMS R.T. = 0.30; [M+H]⁺ = 173.11。 Step A: 6- (Pyridin-4-yl) pyrimidin-4-amine

6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol), pyridin-4-ylboronic acid (0.384 g, 3.13 mmol), Na ₂ CO ₃ (0.795 g, 7.50 mmol) and bis chloride (Triphenylphosphine) palladium (II) (0.035 g, 0.050 mmol) was suspended in a mixture of DME / EtOH / water (15: 2: 3 ml). The mixture was heated in a microwave synthesizer at 125 ° C. for 20 minutes and concentrated. The residue was purified by chromatography on silica gel (5-25% [9: 1 methanol: ammonium hydroxide] -ethyl acetate) to give 6- (pyridin-3-yl) pyrimidin-4-amine as an off-white solid. (0.15 g, 0.871 mmol, 35% yield).
LCMS RT = 0.30; [M + H] ⁺ = 173.11.

ジクロロメタン／Ｎ,Ｎ−ジメチルホルムアミド中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.６０１ｇ, ２.５９mmol)の溶液に、室温で、６−(ピリジン−４−イル)ピリミジン−４−アミン(０.２９７ｇ, １.７２５mmol)を加えた。橙色の混合物を６０℃で１８時間加熱した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物をシリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、４−イソチオシアナト−６−(ピリジン−４−イル)ピリミジンを橙色の固体として得た(０.０５５ｇ, ０.２５７mmol, 収率１５％)。
LCMS R.T. = 1.46; [M+H]⁺ = 215.09。 Step B: 4-Isothiocyanato-6- (pyridin-4-yl) pyrimidine

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) was added. The orange mixture was heated at 60 ° C. for 18 hours. LC / MS showed the desired product peak as the major peak. The deep orange mixture was purified by chromatography on silica gel (0-40% ethyl acetate-hexane) to give 4-isothiocyanato-6- (pyridin-4-yl) pyrimidine as an orange solid (0.055 g, 0 .257 mmol, 15% yield).
LCMS RT = 1.46; [M + H] ⁺ = 215.09.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.０５９ｇ, ０.２５７mmol)に、Ｃｓ_２ＣＯ_３(０.２０９ｇ, ０.６４２mmol)および４−イソチオシアナト−６−(ピリジン−４−イル)ピリミジン(０.０５５ｇ, ０.２５７mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１２ml, ０.７７mmol)を加え、混合物を室温で１８時間撹拌し続けた。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(０〜１０％ [９：１ メタノール：水酸化アンモニウム]−酢酸エチル)、(Ｒ)−Ｎ−(６−(ピリジン−４−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色のフィルム状物質として得た(０.０１４ｇ, ０.０４mmol, 収率１６％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.85 (1 H, d), 8.67 (2 H, dd), 8.04 (2 H, dd), 7.34 (1 H, br. s.), 4.06 (1 H, d), 3.76 (1 H, d), 3.23 (1 H, d), 3.10 (1 H, d), 2.70 - 2.99 (4 H, m), 2.01 - 2.22 (2 H, m), 1.53 - 1.86 (3 H, m). LCMS R.T. = 0.42; [M+H]⁺ = 337.14。 Step C: (R) -N- (6- (Pyridin-4-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.059 g, 0.257 mmol) in N, N-dimethylformamide (15 ml) was added to Cs ₂ CO ₃ (0.209 g, 0 .642 mmol) and 4-isothiocyanato-6- (pyridin-4-yl) pyrimidine (0.055 g, 0.257 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-diisopropylcarbodiimide (0.12 ml, 0.77 mmol) was added and the mixture was kept stirring at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (0-10% [9: 1 methanol: ammonium hydroxide] -ethyl acetate), (R) -N- (6- (pyridin-4-yl) pyrimidine- 4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow film (0.014 g, 0.04 mmol). , Yield 16%).
¹ H NMR (400 MHz, MeOD) δ ppm 8.85 (1 H, d), 8.67 (2 H, dd), 8.04 (2 H, dd), 7.34 (1 H, br.s.), 4.06 (1 H , d), 3.76 (1 H, d), 3.23 (1 H, d), 3.10 (1 H, d), 2.70-2.99 (4 H, m), 2.01-2.22 (2 H, m), 1.53- 1.86 (3 H, m). LCMS RT = 0.42; [M + H] ⁺ = 337.14.

Example 247
(R) -6- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) -2-methylnicotinonitrile

ジクロロメタン(２０ml)中の６−アミノ−２−メチルニコチノニトリル(０.４１ｇ, ３.０８mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.７５ｇ, ３.２３mmol)を加えた。反応物を４０℃で３時間撹拌した。反応物を室温まで冷却した。粗製の物質をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、６−イソチオシアナト−２−メチルニコチノニトリルを得た(０.５２ｇ, ２.９７mmol, 収率９６％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.36 (d, J=8.24 Hz, 1 H), 7.39 (d, J=8.24 Hz, 1 H), 2.65 (s, 3 H). MS (LC/MS) R.T. = 2.09; [M+H]⁺ = 176.0。 Step A: 6-Isothiocyanato-2-methylnicotinonitrile

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) was added. The reaction was stirred at 40 ° C. for 3 hours. The reaction was cooled to room temperature. The crude material was purified by chromatography (Biotage, 25-100% ethyl acetate / hexane) to give 6-isothiocyanato-2-methylnicotinonitrile (0.52 g, 2.97 mmol, 96% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.36 (d, J = 8.24 Hz, 1 H), 7.39 (d, J = 8.24 Hz, 1 H), 2.65 (s, 3 H). MS (LC / MS) RT = 2.09; [M + H] ⁺ = 176.0.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の６−イソチオシアナト−２−メチルニコチノニトリル(０.２５ｇ, １.４３mmol)に、トリエチルアミン(０.５ml, ３.６６６mmol)および３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.３３ｇ, １.４６mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製の尿素をクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.６７ml, ４.２８mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、真空で濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−６−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)−２−メチルニコチノニトリルを白色の粉末として得た(０.０９ｇ, ０.３mmol, 収率２１％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 9.11 (s, 1 H), 7.87 (d, J=7.93 Hz, 2 H), 6.69 (s, 1 H), 3.89 (d, J=10.38 Hz, 2 H), 3.63 (d, J=10.38 Hz, 3 H), 3.00 (s, 5 H), 2.72 - 2.80 (m, 4 H), 2.64 - 2.69 (m, 5 H), 2.60 (s, 7 H), 2.00 (d, J=2.14 Hz, 3 H), 1.91 (s, 1 H), 1.87 (s, 2 H), 1.58 (s, 5 H), 1.42 - 1.50 (m, 2 H). MS (LC/MS) R.T. = 0.48; [M+H]⁺ = 298.13。 Step B: (R) -6- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) -2-methylnicotinonitrile

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) quinuclidine. -3-ol dihydrochloride (0.33 g, 1.46 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (20 ml) and N, N′-diisopropylcarbodiimide (0.67 ml, 4.28 mmol). The reaction was heated at 70 ° C. for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -6- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-ylamino) -2-methylnicotinonitrile was obtained as a white powder (0.09 g, 0.3 mmol, 21% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 9.11 (s, 1 H), 7.87 (d, J = 7.93 Hz, 2 H), 6.69 (s, 1 H), 3.89 (d, J = 10.38 Hz , 2 H), 3.63 (d, J = 10.38 Hz, 3 H), 3.00 (s, 5 H), 2.72-2.80 (m, 4 H), 2.64-2.69 (m, 5 H), 2.60 (s, 7 H), 2.00 (d, J = 2.14 Hz, 3 H), 1.91 (s, 1 H), 1.87 (s, 2 H), 1.58 (s, 5 H), 1.42-1.50 (m, 2 H) MS (LC / MS) RT = 0.48; [M + H] ⁺ = 298.13.

Example 248
(R) -6- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) -2,4-dimethylnicotinonitrile

ジクロロメタン(２０ml)中の６−アミノ−２,４−ジメチルニコチノニトリル(０.１４ｇ, ０.９５mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.２３ｇ, ０.１mmol)を加えた。反応物を４０℃で３時間撹拌した。反応物を室温まで冷却した。粗製の物質をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、６−イソチオシアナト−２,４−ジメチルニコチノニトリルを得た(０.１５ｇ, ０.７９mmol, 収率８３％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 7.37 (s, 1 H), 2.63 (s, 3 H). MS (LC/MS) R.T. = 2.40; [M+H]⁺ = 190。 Step A: 6-Isothiocyanato-2,4-dimethylnicotinonitrile

6-Amino-2,4-dimethylnicotinonitrile (0.14 g, 0.95 mmol) in dichloromethane (20 ml) was added to 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.23 g, 0.1 mmol) was added. The reaction was stirred at 40 ° C. for 3 hours. The reaction was cooled to room temperature. The crude material was purified by chromatography (Biotage, 25-100% ethyl acetate / hexane) to give 6-isothiocyanato-2,4-dimethylnicotinonitrile (0.15 g, 0.79 mmol, 83% yield). ).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 7.37 (s, 1 H), 2.63 (s, 3 H). MS (LC / MS) RT = 2.40; [M + H] ⁺ = 190.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の６−イソチオシアナト−２,４−ジメチルニコチノニトリル(０.０９ｇ, ０.４８mmol)に、トリエチルアミン(０.１７ml, １.１９mmol)および３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１１ｇ, ０.４９mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.２２ml, １.４３mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、真空で濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−６−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)−２,４−ジメチルニコチノニトリルを白色の粉末として得た(０.１０ｇ, ０.３２mmol, 収率６６％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 9.08 (s, 1 H), 6.60 (s, 1 H), 3.88 (d, J=10.38 Hz, 1 H), 3.61 (d, J=10.38 Hz, 1 H), 2.98 (s, 2 H), 2.70 - 2.79 (m, 2 H), 2.63 - 2.69 (m, 2 H), 2.55 - 2.60 (m, 4 H), 2.31 - 2.39 (m, 4 H), 1.99 (s, 1 H), 1.89 (s, 1 H), 1.54 - 1.62 (m, 2 H), 1.41 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 0.78; [M+H]⁺ = 312.1。 Step B: (R) -6- (4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-ylamino) -2,4-dimethylnicotinonitrile

6-isothiocyanato-2,4-dimethylnicotinonitrile (0.09 g, 0.48 mmol) in N, N-dimethylformamide (20 ml) was added to triethylamine (0.17 ml, 1.19 mmol) and 3- (aminomethyl). Quinuclidin-3-ol dihydrochloride (0.11 g, 0.49 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (20 ml) and N, N′-diisopropylcarbodiimide (0.22 ml, 1.43 mmol). The reaction was heated at 70 ° C. for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -6- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-ylamino) -2,4-dimethylnicotinonitrile was obtained as a white powder (0.10 g, 0.32 mmol, 66% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 9.08 (s, 1 H), 6.60 (s, 1 H), 3.88 (d, J = 10.38 Hz, 1 H), 3.61 (d, J = 10.38 Hz , 1 H), 2.98 (s, 2 H), 2.70-2.79 (m, 2 H), 2.63-2.69 (m, 2 H), 2.55-2.60 (m, 4 H), 2.31-2.39 (m, 4 H), 1.99 (s, 1 H), 1.89 (s, 1 H), 1.54-1.62 (m, 2 H), 1.41-1.49 (m, 1 H). MS (LC / MS) RT = 0.78; [ M + H] ⁺ = 312.1.

Example 249
(R) -N- (6-Phenylpyridazin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

３−イソチオシアナト−６−フェニルピリダジンを、実施例２３の工程Ｂの方法によって合成した。１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分でフラッシュクロマトグラフィーを行い、４２０mgを得た(収率４９％)。
LCMS: RT = 2.17分, MH⁺ = 214.06。 Step A: 3-isothiocyanato-6-phenylpyridazine

3-isothiocyanato-6-phenylpyridazine was synthesized by the method of Example 23, Step B. Flash chromatography on a 120 g silica gel cartridge with 0-25% EtOAc in hexanes, 25 min, 35 ml / min yielded 420 mg (49% yield).
LCMS: RT = 2.17 min, MH ⁺ = 214.06.

(Ｒ)−Ｎ−(６−フェニルピリダジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ｂの方法によって合成した。１６０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜４％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分、４０ml/分で、フラッシュクロマトグラフィーを行い、６７mgの(Ｒ)−Ｎ−(６−フェニルピリダジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(収率１７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.61 (1 H, br. s.), 7.95 - 7.99 (2 H, m), 7.74 (1 H, d, J=9.32 Hz), 7.39 - 7.52 (3 H, m), 7.22 (1 H, partial d), 3.97 (1 H, d, J=9.32 Hz), 3.64 (1 H, d, J=9.32 Hz), 3.37 (1 H, dd, J=14.73, 1.38 Hz), 2.69 - 3.06 (5 H, m), 2.16 - 2.26 (1 H, m), 2.14 (1 H, br. s.), 1.66 - 1.79 (1 H, m), 1.45 - 1.60 (2 H, m)
¹H NMR (400 MHz, MeOD) δ ppm 7.87 - 8.02 (3 H, m), 7.44 - 7.55 (3 H, m), 7.13 - 7.29 (1 H, m), 4.05 (1 H, d, J=9.82 Hz), 3.74 (1 H, d, J=10.07 Hz), 3.17 (2 H, dd, J=49.35, 14.60 Hz), 2.73 - 3.04 (4 H, m), 2.16 (2 H, br. s.), 1.55 - 1.85 (3 H, m)
LCMS: RT = 0.82分, MH^- = 334.2, MH⁺ = 336.2。 Step B: (R) -N- (6-Phenylpyridazin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Phenylpyridazin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 23. It was synthesized by the method of Step B. Flash chromatography was performed on a 160 g silica gel cartridge with 1-4% [9: 1 MeOH / NH ₄ OH] in CHCl ₃ , 50 min, 40 ml / min, and 67 mg (R) -N- (6- Phenylpyridazin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (yield 17%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.61 (1 H, br.s.), 7.95-7.99 (2 H, m), 7.74 (1 H, d, J = 9.32 Hz), 7.39-7.52 ( 3 H, m), 7.22 (1 H, partial d), 3.97 (1 H, d, J = 9.32 Hz), 3.64 (1 H, d, J = 9.32 Hz), 3.37 (1 H, dd, J = 14.73, 1.38 Hz), 2.69-3.06 (5 H, m), 2.16-2.26 (1 H, m), 2.14 (1 H, br.s.), 1.66-1.79 (1 H, m), 1.45-1.60 (2 H, m)
¹ H NMR (400 MHz, MeOD) δ ppm 7.87-8.02 (3 H, m), 7.44-7.55 (3 H, m), 7.13-7.29 (1 H, m), 4.05 (1 H, d, J = 9.82 Hz), 3.74 (1 H, d, J = 10.07 Hz), 3.17 (2 H, dd, J = 49.35, 14.60 Hz), 2.73-3.04 (4 H, m), 2.16 (2 H, br. S .), 1.55-1.85 (3 H, 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] octane] -2-amine

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の５−ブロモピラジン−２−アミン(２ｇ, １１.４９mmol)の溶液に、ナトリウム チオメトキシド(１.６１１ｇ, ２２.９９mmol)を加えた。混合物を、窒素下、１００℃で１８時間加熱し、濃縮した。残渣を水で処理し、混合物をジクロロメタンで抽出した。合わせた有機物を硫酸ナトリウムで乾燥させ、濾過し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(０〜１０％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、６−(ピリジン−３−イル)ピリミジン−４−アミンを黄色の固体として得た(０.１５ｇ, ０.８７１mmol, 収率３５％)。
LCMS R.T. = 0.91; [M+H]⁺ = 141.89。 Step A: 5- (Methylthio) pyrazin-2-amine

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 heated at 100 ° C. under nitrogen for 18 hours and concentrated. The residue was treated with water and the mixture was extracted with dichloromethane. The combined organics were dried over sodium sulfate, filtered and concentrated. The residue was purified by chromatography on silica gel (0-10% 9: 1 methanol: ammonium hydroxide-ethyl acetate) to give 6- (pyridin-3-yl) pyrimidin-4-amine as a yellow solid (0 .15 g, 0.871 mmol, 35% yield).
LCMS RT = 0.91; [M + H] ⁺ = 141.89.

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１.０６９ｇ, ４.６０mmol)の溶液に、室温で、５−(メチルチオ)ピラジン−２−アミン(０.５０ｇ, ３.５４mmol)を加えた。反応物を室温で１８時間撹拌した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物をシリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、２−イソチオシアナト−５−(メチルチオ)ピラジンを橙色の油状物として得た(０.５４５ｇ, ０.２５７mmol, 収率８４％)。
LCMS R.T. = 2.65; [M+H]⁺ = 184.02。 Step B: 2-isothiocyanato-5- (methylthio) pyrazine

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (1.069 g, 4.60 mmol) in dichloromethane at room temperature, 5- (methylthio) pyrazin-2-amine (0.50 g, 3.54 mmol) was added. 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 chromatography on silica gel (0-40% ethyl acetate-hexane) to give 2-isothiocyanato-5- (methylthio) pyrazine as an orange oil (0.545 g, 0.257 mmol, Yield 84%).
LCMS RT = 2.65; [M + H] ⁺ = 184.02.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.３７５ｇ, １.６３７mmol)に、Ｃｓ_２ＣＯ_３(１.３３３ｇ, ４.０９mmol)および２−イソチオシアナト−５−(メチルチオ)ピラジン(０.３ｇ, １.６３７mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.７６５ml, ４.９mmol)を加え、混合物を室温で１８時間撹拌し続けた。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(５−(メチルチオ)ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.０１４ｇ, ０.０４mmol, 収率１６％)。
融点 155〜60℃. ¹H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, d), 8.06 (1 H, s), 3.96 (1 H, d), 3.66 (1 H, d), 3.20 (1 H, d), 3.08 (1 H, d), 2.87 - 2.96 (2 H, m), 2.72 - 2.82 (2 H, m), 2.52 (3 H, s), 2.00 - 2.19 (2 H, m), 1.51 - 1.81 (3 H, m). LCMS R.T. = 1.01; [M+H]⁺ = 306.12。 Step C: (R) -N- (5- (methylthio) pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.375 g, 1.637 mmol) in N, N-dimethylformamide (15 ml) was added to Cs ₂ CO ₃ (1.333 g, 4 0.09 mmol) and 2-isothiocyanato-5- (methylthio) pyrazine (0.3 g, 1.637 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.765 ml, 4.9 mmol) was added and the mixture continued to stir at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (5- (methylthio) pyrazin-2-yl) -4H -1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.014 g, 0.04 mmol, 16% yield).
Melting point 155-60 ° C. ¹ H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, d), 8.06 (1 H, s), 3.96 (1 H, d), 3.66 (1 H, d), 3.20 (1 H, d), 3.08 (1 H, d), 2.87-2.96 (2 H, m), 2.72-2.82 (2 H, m), 2.52 (3 H, s), 2.00-2.19 (2 H, m), 1.51-1.81 (3 H, m). LCMS RT = 1.01; [M + H] ⁺ = 306.12.

Example 251
(R) -N- (5,6-Dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

クロロホルム(２５ml)中の、J. Org. Chem 2005, 70, 1771の通りに合成したＮ−(６−クロロピリジン−２−イル)ピバル酸アミド(１.０２ｇ, ４.８０mmol)の溶液に、１−クロロピロリジン−２,５−ジオン(０.６２ｇ, ４.６７mmol)を加え、混合物を油浴中で３時間還流した。それを一夜かけて室温まで冷却した。反応混合物を真空で蒸発させ、ＤＭＦ(１５ml)に再度溶解した。さらに４８０mgの１−クロロピロリジン−２,５−ジオンを加え、得られた溶液を油浴中で９５〜１００℃で一夜加熱し、再度室温まで冷却した。溶媒を真空で除去し、残渣を水と酢酸エチルの層間に分配した。水相を酢酸エチルで２回以上洗浄し、合わせた有機相を塩水で洗浄し、硫酸マグネシウムで乾燥させ、真空で蒸発させた。ＴＬＣ(１０％ 酢酸エチル／ヘキサン)により、Ｒｆ＝０.６の濃いスポットと、Ｒｆ＝０.４および０.２のより小さいスポットが示された。この物質を５〜１０％ 酢酸エチル／ヘキサンでBiotageに流し、Ｒｆ＝０.６のフラクションを集めて、７９０mg(６６％)の白色の固体であるＮ−(５,６−ジクロロピリジン−２−イル)ピバル酸アミドを得た。
¹H NMR (500 MHz, CDCl₃) δ ppm 8.17 (s, 1 H), 7.96 (s, 1 H), 7.72 (s, 1 H), 1.31 (s, 10 H). MS (LC/MS) R.T. = 1.85; [M+H]⁺ = 248.8。 Step A: N- (5,6-dichloropyridin-2-yl) pivalic acid amide

To a solution of N- (6-chloropyridin-2-yl) pivalic acid amide (1.02 g, 4.80 mmol) synthesized as in J. Org. Chem 2005, 70 , 1771 in chloroform (25 ml), 1-Chloropyrrolidine-2,5-dione (0.62 g, 4.67 mmol) was added and the mixture was refluxed in an oil bath for 3 hours. It was cooled to room temperature overnight. The reaction mixture was evaporated in vacuo and redissolved in DMF (15 ml). An additional 480 mg of 1-chloropyrrolidine-2,5-dione was added and the resulting solution was heated in an oil bath at 95-100 ° C. overnight and again cooled 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 / hexanes) showed a dark spot with Rf = 0.6 and smaller spots with Rf = 0.4 and 0.2. This material was flushed into the Biotage with 5-10% ethyl acetate / hexanes and the Rf = 0.6 fraction was collected to yield 790 mg (66%) of a white solid N- (5,6-dichloropyridine-2- Yl) pivalic acid amide was obtained.
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.17 (s, 1 H), 7.96 (s, 1 H), 7.72 (s, 1 H), 1.31 (s, 10 H). MS (LC / MS) RT = 1.85; [M + H] ⁺ = 248.8.

Ｎ−(５,６−ジクロロピリジン−２−イル)ピバル酸アミド(７９０mg, ３.２０mmol)、塩酸 ３７％(１.２５ml)、水(１.２５ml)およびＥｔＯＨ(３ml)の混合物を、油浴中で８５〜９０℃で４時間加熱した。ＬＣＭＳにより、生成物への変換がほとんど完了したことが示された。反応物を室温まで冷却し、反応混合物を少量になるまで蒸発させ、分液漏斗に移し、それを水性炭酸ナトリウムと酢酸エチルの層間に分配した。層を分離し、水相を酢酸エチルで再度洗浄し、合わせた有機相を塩水で洗浄し、ＭｇＳＯ_４で乾燥させ、濾過し、蒸発させ、白色の固体を得た。該物質を２０％ 酢酸エチル／ヘキサンでBiotage カラムに流し、主要な成分を集めて、５,６−ジクロロピリジン−２−アミンを白色の固体として得た(０.４９ｇ, ２.９８mmol, ９３％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 7.44 (d, J=8.55 Hz, 1 H), 6.36 (d, J=8.24 Hz, 1 H), 4.58 (s, 2 H). MS (LC/MS) R.T. = 1.28; [M+H]⁺ = 164.8。 Step B: 5,6-Dichloropyridin-2-amine

A mixture of N- (5,6-dichloropyridin-2-yl) pivalic acid amide (790 mg, 3.20 mmol), hydrochloric acid 37% (1.25 ml), water (1.25 ml) and EtOH (3 ml) was added to the oil. Heated in a bath at 85-90 ° C. for 4 hours. LCMS showed almost complete conversion to product. The reaction was cooled to room temperature and the reaction mixture was evaporated to a small volume and transferred to a separatory funnel which 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 ₄ , filtered and evaporated to give a white solid. The material was run through a Biotage column with 20% ethyl acetate / hexanes and the major components were collected to give 5,6-dichloropyridin-2-amine as a white solid (0.49 g, 2.98 mmol, 93% ).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.44 (d, J = 8.55 Hz, 1 H), 6.36 (d, J = 8.24 Hz, 1 H), 4.58 (s, 2 H). MS (LC / MS) RT = 1.28; [M + H] ⁺ = 164.8.

ジクロロメタン(２５ml)中の５,６−ジクロロピリジン−２−アミン(０.４７ｇ, ２.８８mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.６８ｇ, ２.９４mmol)を加えた。反応物を４０℃で３時間撹拌し、室温まで冷却した。粗製の物質をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、２,３−ジクロロ−６−イソチオシアナトピリジンを白色の粉末として得た(０.４８ｇ, ２.３４mmol, 収率８１％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.26 (d, J=8.55 Hz, 1 H), 7.47 (d, J=8.24 Hz, 1 H). MS (LC/MS) R.T. = 2.83; [M+H]⁺ = 204.8。 Step C: 5,6-Dichloro-2-isothiocyanatopyridine

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) was added. The reaction was stirred at 40 ° C. for 3 hours and cooled to room temperature. The crude material was purified by chromatography (Biotage, 25-100% ethyl acetate / hexane) to give 2,3-dichloro-6-isothiocyanatopyridine as a white powder (0.48 g, 2.34 mmol, Yield 81%).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.26 (d, J = 8.55 Hz, 1 H), 7.47 (d, J = 8.24 Hz, 1 H). MS (LC / MS) RT = 2.83; [ M + H] ⁺ = 204.8.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の２,３−ジクロロ−６−イソチオシアナトピリジン(０.４７ｇ, ２.２９mmol)に、トリエチルアミン(０.８ml, ５.７mmol)および３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.５４ｇ, ２.３４mmol)を室温で加えた。反応物を７０℃で２時間撹拌し、室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(１.０７ml, ６.８８mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、真空で濃縮した。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(５,６−ジクロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.３６ｇ, １.０８mmol, 収率４７％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.31 (d, J=1.22 Hz, 1 H), 7.84 (s, 1 H), 6.80 (s, 1 H), 3.85 (d, J=10.07 Hz, 1 H), 3.57 (d, J=10.38 Hz, 2 H), 2.98 (s, 3 H), 2.69 - 2.78 (m, 3 H), 2.65 (t, J=7.78 Hz, 3 H), 2.00 (s, 2 H), 1.86 (s, 2 H), 1.58 (dd, J=7.48, 2.90 Hz, 2 H), 1.56 (s, 1 H), 1.41 - 1.49 (m, 2 H). MS (LC/MS) R.T. = 0.81; [M+H]⁺ = 327.1。 Step D: (R) -N- (5,6-Dichloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

2,3-Dichloro-6-isothiocyanatopyridine (0.47 g, 2.29 mmol) in N, N-dimethylformamide (20 ml) was added to triethylamine (0.8 ml, 5.7 mmol) and 3- (aminomethyl). Quinuclidin-3-ol dihydrochloride (0.54 g, 2.34 mmol) was added at room temperature. The reaction was stirred at 70 ° C. for 2 hours, cooled to room temperature and concentrated in vacuo. The crude urea was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (20 ml) and N, N′-diisopropylcarbodiimide (1.07 ml, 6.88 mmol). The reaction was heated at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH), (R) -N- (5,6-dichloropyridin-2-yl) -4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.36 g, 1.08 mmol, 47% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.31 (d, J = 1.22 Hz, 1 H), 7.84 (s, 1 H), 6.80 (s, 1 H), 3.85 (d, J = 10.07 Hz , 1 H), 3.57 (d, J = 10.38 Hz, 2 H), 2.98 (s, 3 H), 2.69-2.78 (m, 3 H), 2.65 (t, J = 7.78 Hz, 3 H), 2.00 (s, 2 H), 1.86 (s, 2 H), 1.58 (dd, J = 7.48, 2.90 Hz, 2 H), 1.56 (s, 1 H), 1.41-1.49 (m, 2 H). MS ( LC / MS) RT = 0.81; [M + H] ⁺ = 327.1.

Example 252
(R) -N- (4,5-dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

ジクロロメタン(２５ml)中の４,５−ジクロロピリジン−２−アミン(０.２５ｇ, １.５３mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.３６ｇ, １.５６mmol)を加えた。反応物を４０℃で３時間撹拌し、室温まで冷却した。粗製の物質をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、４,５−ジクロロ−２−イソチオシアナトピリジンを黄色の粉末として得た(０.２６ｇ, １.２７mmol, 収率８３％)。生成物を、次の工程に直接用いた。 Step A: 4,5-dichloro-2-isothiocyanatopyridine

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) was added. The reaction was stirred at 40 ° C. for 3 hours and cooled to room temperature. The crude material was purified by chromatography (Biotage, 25-100% ethyl acetate / hexane) to give 4,5-dichloro-2-isothiocyanatopyridine as a yellow powder (0.26 g, 1.27 mmol, Yield 83%). The product was used directly in the next step.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の４,５−ジクロロ−２−イソチオシアナトピリジン(０.２５ｇ, １.２２mmol)に、トリエチルアミン(０.４３ml, ３.０５mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２９ｇ, １.２４mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.５７ml, ３.６６mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(４,５−ジクロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０９ｇ, ０.２７mmol, 収率２２％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.81 (s, 1 H), 8.32 (d, J=6.10 Hz, 1 H), 7.03 (s, 1 H), 3.83 (d, J=9.46 Hz, 1 H), 3.57 (d, J=9.77 Hz, 1 H), 2.98 (s, 2 H), 2.71 - 2.79 (m, 2 H), 2.65 (t, J=7.78 Hz, 2 H), 1.99 (s, 1 H), 1.86 (s, 1 H), 1.53 - 1.61 (m, 2 H), 1.41 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 0.78; [M+H]⁺ = 327.0。 Step B: (R) -N- (4,5-dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

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) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (20 ml) and N, N′-diisopropylcarbodiimide (0.57 ml, 3.66 mmol). The reaction was heated at 70 ° C. for 2 hours. The reaction was cooled to room temperature and concentrated to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -N- (4,5-dichloropyridin-2-yl) -4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.09 g, 0.27 mmol, 22% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.81 (s, 1 H), 8.32 (d, J = 6.10 Hz, 1 H), 7.03 (s, 1 H), 3.83 (d, J = 9.46 Hz , 1 H), 3.57 (d, J = 9.77 Hz, 1 H), 2.98 (s, 2 H), 2.71-2.79 (m, 2 H), 2.65 (t, J = 7.78 Hz, 2 H), 1.99 (s, 1 H), 1.86 (s, 1 H), 1.53-1.61 (m, 2 H), 1.41-1.49 (m, 1 H). MS (LC / MS) RT = 0.78; [M + H] ⁺ = 327.0.

Example 253
(R) -N- (5-Chloro-4-methylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ジクロロメタン(２５ml)中の５−クロロ−４−メチルピリジン−２−アミン(０.４１ｇ, ２.８８mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.７０ｇ, ３.０mmol)を加えた。反応物を４０℃で３時間撹拌した。反応物を室温まで冷却した。粗製の混合物をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、５−クロロ−２−イソチオシアナト−４−メチルピリジンを得た(０.４５ｇ, ２.４４mmol, 収率８５％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.45 (s, 1 H), 7.47 (s, 1 H), 2.37 (s, 3 H). LC/MS RT=2.79; [M+H]⁺ = 184.9。 Step A: 5-Chloro-2-isothiocyanato-4-methylpyridine

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) was added. The reaction was stirred at 40 ° 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 give 5-chloro-2-isothiocyanato-4-methylpyridine (0.45 g, 2.44 mmol, 85% yield). ).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.45 (s, 1 H), 7.47 (s, 1 H), 2.37 (s, 3 H). LC / MS RT = 2.79; [M + H] ⁺ = 184.9.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の５−クロロ−２−イソチオシアナト−４−メチルピリジン(０.３７ｇ, ２.０mmol)に、トリエチルアミン(０.７ml, ５.０mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(実施例１７の工程Ｂより)(０.４７ｇ, ２.０mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.９４ml, ６.０mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(５−クロロ−４−メチルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.１９ｇ, ０.６１mmol, 収率３０.３％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.79 (s, 1 H), 8.08 - 8.15 (m, 2 H), 6.78 (s, 1 H), 3.82 (d, J=8.55 Hz, 2 H), 3.55 (d, J=10.38 Hz, 2 H), 2.93 - 3.02 (m, 5 H), 2.71 - 2.80 (m, 5 H), 2.66 (t, J=7.63 Hz, 4 H), 2.23 - 2.29 (m, 7 H), 1.94 - 2.02 (m, 2 H), 1.92 (s, 1 H), 1.86 (s, 2 H), 1.53 - 1.62 (m, 5 H), 1.41 - 1.49 (m, J=12.55, 9.88, 7.02, 2.29 Hz, 2 H). MS (LC/MS) R.T. = 0.72; [M+H]⁺ = 307.1。 Step B: (R) -N- (5-Chloro-4-methylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

5-Chloro-2-isothiocyanato-4-methylpyridine (0.37 g, 2.0 mmol) in N, N-dimethylformamide (20 ml) was added to 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) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (20 ml) and N, N′-diisopropylcarbodiimide (0.94 ml, 6.0 mmol). The reaction was heated at 70 ° C. for 2 hours. The reaction was cooled to room temperature and concentrated to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -N- (5-chloro-4-methylpyridin-2-yl) -4H. -1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.19 g, 0.61 mmol, yield 30.3%). .
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.79 (s, 1 H), 8.08-8.15 (m, 2 H), 6.78 (s, 1 H), 3.82 (d, J = 8.55 Hz, 2 H ), 3.55 (d, J = 10.38 Hz, 2 H), 2.93-3.02 (m, 5 H), 2.71-2.80 (m, 5 H), 2.66 (t, J = 7.63 Hz, 4 H), 2.23- 2.29 (m, 7 H), 1.94-2.02 (m, 2 H), 1.92 (s, 1 H), 1.86 (s, 2 H), 1.53-1.62 (m, 5 H), 1.41-1.49 (m, J = 12.55, 9.88, 7.02, 2.29 Hz, 2 H). MS (LC / MS) RT = 0.72; [M + H] ⁺ = 307.1.

Example 254
(R) -N- (6-Chloropyridazin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

３−クロロ−６−イソチオシアナトピリダジンを、実施例２１８の工程Ｄの方法によって合成した。１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分でフラッシュクロマトグラフィーを行い、２１３mgを得た(収率３１％)。
LCMS: RT = 1.25分, MH⁺ = 172.00。 Step A: 3-Chloro-6-isothiocyanatopyridazine

3-Chloro-6-isothiocyanatopyridazine was synthesized by the method of Example 218, Step D. Flash chromatography on a 120 g silica gel cartridge with 0-25% EtOAc in hexanes, 25 min, 35 ml / min yielded 213 mg (31% yield).
LCMS: RT = 1.25 min, MH ⁺ = 172.00.

(Ｒ)−Ｎ−(６−クロロピリダジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２１８の工程Ｅの方法によって合成した。１６０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜３％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分、４０ml/分でフラッシュクロマトグラフィーを行い、２９mgを得た(収率８％)。
1H NMR (400 MHz, CDCl₃) δ ppm 9.27 (1 H, br. s.), 7.28 (1 H, d, J=9.07 Hz), 7.10 (1 H, d, J=9.07 Hz), 3.95 (1 H, d, J=9.57 Hz), 3.62 (1 H, d, J=9.57 Hz), 3.34 (1 H, dd, J=14.98, 1.64 Hz), 2.67 - 3.04 (5 H, m), 2.14 - 2.21 (1 H, m), 2.12 (1 H, br. s.), 1.65 - 1.79 (1 H, m), 1.45 - 1.61 (2 H, m). LCMS: RT = 0.62分, MH^- = 292.1, MH⁺ = 294.1。 Step B: (R) -N- (6-Chloropyridazin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) —N- (6-Chloropyridazin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 218. It was synthesized by the method of Step E. Flash chromatography on a 160 g silica gel cartridge with 1-3% [9: 1 MeOH / NH ₄ OH] in CHCl ₃ , 50 min, 40 ml / min, gave 29 mg (8% yield).
1H NMR (400 MHz, CDCl ₃ ) δ ppm 9.27 (1 H, br.s.), 7.28 (1 H, d, J = 9.07 Hz), 7.10 (1 H, d, J = 9.07 Hz), 3.95 ( 1 H, d, J = 9.57 Hz), 3.62 (1 H, d, J = 9.57 Hz), 3.34 (1 H, dd, J = 14.98, 1.64 Hz), 2.67-3.04 (5 H, m), 2.14 - 2.21 (1 H, m) , 2.12, 1.65 - 1.79 (1 H, m), 1.45 - 1.61 (2 H, m) LCMS (1 H, br s..):. 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] octane] -2-amine

３−ブロモ−６−イソチオシアナトピリダジンを、実施例２１８の工程Ｄの方法によって合成した。１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２５％ ＥｔＯＡｃ、２５分、３５ml/分でフラッシュクロマトグラフィーを行い、３６４mgを得た(収率４２％)。
LCMS: RT = 1.34分, MH⁺ = 215.92。 Step A: 3-Bromo-6-isothiocyanatopyridazine

3-Bromo-6-isothiocyanatopyridazine was synthesized by the method of Example 218, Step D. Flash chromatography on a 120 g silica gel cartridge with 0-25% EtOAc in hexanes, 25 min, 35 ml / min gave 364 mg (42% yield).
LCMS: RT = 1.34 min, MH ⁺ = 215.92.

(Ｒ)−Ｎ−(６−ブロモピリダジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２１８の工程Ｅの方法によって合成した。１６０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中１〜３％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]、５０分、４０ml/分でフラッシュクロマトグラフィーを行い、２１１mgを得た(収率３７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.23 (1 H, br. s.), 7.37 (1 H, d, J=9.07 Hz), 6.97 (1 H, d, J=9.07 Hz), 3.92 (1 H, d, J=9.57 Hz), 3.59 (1 H, d, J=9.82 Hz), 3.29 (1 H, dd, J=14.98, 1.64 Hz), 2.63 - 2.99 (5 H, m), 2.04 - 2.19 (2 H, m), 1.59 - 1.74 (1 H, m), 1.39 - 1.58 (2 H, m). LCMS: RT= 0.64分, MH^- 336.1, MH⁺ 338.0。 Step B: (R) -N- (6-Bromopyridazin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Bromopyridazin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 218. It was synthesized by the method of Step E. Flash chromatography on a 160 g silica gel cartridge with 1-3% [9: 1 MeOH / NH ₄ OH] in CHCl ₃ , 50 min, 40 ml / min yielded 211 mg (37% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.23 (1 H, br.s.), 7.37 (1 H, d, J = 9.07 Hz), 6.97 (1 H, d, J = 9.07 Hz), 3.92 (1 H, d, J = 9.57 Hz), 3.59 (1 H, d, J = 9.82 Hz), 3.29 (1 H, dd, J = 14.98, 1.64 Hz), 2.63-2.99 (5 H, m), 2.04-2.19 (2 H, m), 1.59-1.74 (1 H, m), 1.39-1.58 (2 H, m). 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′-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(０.３２４ｇ, ２.５mmol)、４−クロロフェニルボロン酸(０.４８９ｇ, ３.１３mmol)、Ｎａ_２ＣＯ_３(０.７９５ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５０mmol)の混合物を、ＤＭＥ／ＥｔＯＨ／水(１５：２：３ml)の混合物に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(２〜１５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、６−(ピリジン−３−イル)ピリミジン−４−アミンを灰白色の固体として得た(０.３ｇ, ０.８７１mmol, 収率５８.４％)。
LCMS R.T. = 1.42; [M+2H]⁺ = 207.91。 Step A: 6- (Pyridin-4-yl) pyrimidin-4-amine

6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol), 4-chlorophenylboronic acid (0.489 g, 3.13 mmol), Na ₂ CO ₃ (0.795 g, 7.50 mmol) and bis chloride ( A mixture of triphenylphosphine) palladium (II) (0.035 g, 0.050 mmol) was suspended in a mixture of DME / EtOH / water (15: 2: 3 ml). The mixture was heated in a microwave synthesizer at 125 ° C. for 20 minutes and concentrated. The residue was purified by chromatography on silica gel (2-15% 9: 1 methanol: ammonium hydroxide-ethyl acetate) to give 6- (pyridin-3-yl) pyrimidin-4-amine as an off-white solid (0 .3 g, 0.871 mmol, yield 58.4%).
LCMS RT = 1.42; [M + 2H] ⁺ = 207.91.

ジクロロメタン／Ｎ,Ｎ−ジメチルホルムアミド中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.６６６ｇ, ２.８７mmol)の明橙色の溶液に、室温で、６−(４−クロロフェニル)ピリミジン−４−アミン(０.５９ｇ, ２.８７mmol)を加えた。橙色の混合物を６０℃で１８時間加熱した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物をシリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、４−(４−クロロフェニル)−６−イソチオシアナトピリミジンを橙色の油状物として得た(０.３２２ｇ, １.３００mmol, 収率４５％)。
LCMS R.T. = 2.82; [M]⁺ = 248.03。 Step B: 4- (4-Chlorophenyl) -6-isothiocyanatopyrimidine

To a light orange solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.666 g, 2.87 mmol) in dichloromethane / N, N-dimethylformamide at room temperature, 6- (4- Chlorophenyl) pyrimidin-4-amine (0.59 g, 2.87 mmol) was added. The orange mixture was heated at 60 ° C. for 18 hours. LC / MS showed the desired product peak as the major peak. The deep orange mixture was purified by chromatography on silica gel (0-40% ethyl acetate-hexane) to give 4- (4-chlorophenyl) -6-isothiocyanatopyrimidine as an orange oil (0.322 g, 1.300 mmol, 45% yield).
LCMS RT = 2.82; [M] ⁺ = 248.03.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２９８ｇ, １.３００mmol)に、Ｃｓ_２ＣＯ_３(１.０５９ｇ, ３.２５mmol)および４−(４−クロロフェニル)−６−イソチオシアナトピリミジン(０.３２２ｇ, １.３００mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.６０８ml, ３.９０mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(６−(ピリジン−４−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.１０４ｇ, ０.２７６mmol, 収率２１％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.54 (1 H, br. s.), 8.83 (1 H, d), 7.89 - 8.04 (2 H, m), 7.41 - 7.54 (2 H, m), 7.33 (1 H, br. s.), 4.02 (1 H, d), 3.71 (1 H, d), 3.42 (1 H, d), 2.73 - 3.15 (5 H, m), 2.10 - 2.31 (2 H, m), 1.46 - 1.89 (3 H, m). LCMS R.T. = 1.92; [M]⁺ = 370.35。 Step C: (R) -N- (6- (4-Chlorophenyl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.298 g, 1.300 mmol) in N, N-dimethylformamide (15 ml) was added to Cs ₂ CO ₃ (1.059 g, 3 .25 mmol) and 4- (4-chlorophenyl) -6-isothiocyanatopyrimidine (0.322 g, 1.300 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.608 ml, 3.90 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (6- (pyridin-4-yl) pyrimidine-4- Yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.104 g, 0.276 mmol, yield 21). %).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.54 (1 H, br. S.), 8.83 (1 H, d), 7.89-8.04 (2 H, m), 7.41-7.54 (2 H, m) , 7.33 (1 H, br. S.), 4.02 (1 H, d), 3.71 (1 H, d), 3.42 (1 H, d), 2.73-3.15 (5 H, m), 2.10-2.31 ( 2 H, m), 1.46-1.89 (3 H, m). LCMS RT = 1.92; [M] ⁺ = 370.35.

Example 257
(R) -N- (6- (3-Chlorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(０.３２４ｇ, ２.５mmol)、３−クロロフェニルボロン酸(０.４８９ｇ, ３.１３mmol)、Ｎａ_２ＣＯ_３(０.７９５ｇ, ７.５０mmol)および塩化 ビス(トリフェニルホスフィン)パラジウム(II)(０.０３５ｇ, ０.０５０mmol)の混合物を、ＤＭＥ／ＥｔＯＨ／水(１５：２：３ml)の混合物に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮した。残渣をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中３０〜７０％ 酢酸エチル)、６−(３−クロロフェニル)ピリミジン−４−アミンを黄色の固体として得た(０.４７ｇ, ２.２８６mmol, 収率９１％)。
LCMS R.T. = 1.45; [M+2H]⁺ = 208.05。 Step A: 6- (3-Chlorophenyl) pyrimidin-4-amine

6-chloropyrimidin-4-amine (0.324 g, 2.5 mmol), 3-chlorophenylboronic acid (0.489 g, 3.13 mmol), Na ₂ CO ₃ (0.795 g, 7.50 mmol) and bis ( A mixture of triphenylphosphine) palladium (II) (0.035 g, 0.050 mmol) was suspended in a mixture of DME / EtOH / water (15: 2: 3 ml). The mixture was heated in a microwave synthesizer at 125 ° C. for 20 minutes and concentrated. The residue was purified by chromatography on silica gel (30-70% ethyl acetate in hexanes) to give 6- (3-chlorophenyl) pyrimidin-4-amine as a yellow solid (0.47 g, 2.286 mmol, yield). 91%).
LCMS RT = 1.45; [M + 2H] ⁺ = 208.05.

ジクロロメタン／Ｎ,Ｎ−ジメチルホルムアミド中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.４８６ｇ, ２.０９１mmol)の明橙色の溶液に、室温で、６−(３−クロロフェニル)ピリミジン−４−アミン(０.４３ｇ, ２.０９１mmol)を加えた。橙色の混合物を６０℃で１８時間加熱した。ＬＣ／ＭＳにより、望ましい生成物のピークが主要なピークとして示された。深橙色の混合物をシリカゲルのクロマトグラフィーによって精製し(０〜４０％ 酢酸エチル−ヘキサン)、４−(３−クロロフェニル)−６−イソチオシアナトピリミジンを橙色の油状物として得た(０.１２ｇ, ０.４８４mmol, 収率２３％)。
LCMS R.T. = 2.15; [M]⁺ = 248.31。 Step B: 4- (3-Chlorophenyl) -6-isothiocyanatopyrimidine

To a light orange solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.486 g, 2.091 mmol) in dichloromethane / N, N-dimethylformamide at room temperature, 6- (3- Chlorophenyl) pyrimidin-4-amine (0.43 g, 2.091 mmol) was added. The orange mixture was heated at 60 ° C. for 18 hours. LC / MS showed the desired product peak as the major peak. The deep orange mixture was purified by chromatography on silica gel (0-40% ethyl acetate-hexane) to give 4- (3-chlorophenyl) -6-isothiocyanatopyrimidine as an orange oil (0.12 g, 0.484 mmol, 23% yield).
LCMS RT = 2.15; [M] ⁺ = 248.31.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１１１ｇ, ０.４８４mmol)に、Ｃｓ_２ＣＯ_３(０.３９５ｇ, １.２１１mmol)および４−(３−クロロフェニル)−６−イソチオシアナトピリミジン(０.１２ｇ, ０.４８４mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.２２６ml, １.４５３mmol)を加え、混合物を室温で１８時間撹拌し続けた。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％、次に２〜１０％の９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(６−(３−クロロフェニル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.０８６ｇ, ０.２２１mmol, 収率４６％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.82 (1 H, d), 8.05 (1 H, d), 7.93 (1 H, ddd), 7.43 - 7.52 (2 H, m), 7.35 (1 H, br. s.), 4.13 (1 H, d), 3.93 (1 H, d), 3.63 - 3.81 (2 H, m), 3.42 - 3.53 (1 H, m), 3.30 - 3.40 (3 H, m), 2.46 (1 H, d), 2.26 - 2.40 (1 H, m), 1.88 - 2.16 (3 H, m). LCMS R.T. = 1.90; [M]⁺ = 370.28。 Step C: (R) -N- (6- (3-chlorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.111 g, 0.484 mmol) in N, N-dimethylformamide (15 ml) was added to Cs ₂ CO ₃ (0.395 g, 1 0.21 mmol) and 4- (3-chlorophenyl) -6-isothiocyanatopyrimidine (0.12 g, 0.484 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-diisopropylcarbodiimide (0.226 ml, 1.453 mmol) was added and the mixture was kept stirring at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25%, then 2-10% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (6- (3- Chlorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.086 g, 0 .221 mmol, 46% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.82 (1 H, d), 8.05 (1 H, d), 7.93 (1 H, ddd), 7.43-7.52 (2 H, m), 7.35 (1 H, br. s.), 4.13 (1 H, d), 3.93 (1 H, d), 3.63-3.81 (2 H, m), 3.42-3.53 (1 H, m), 3.30-3.40 (3 H, m ), 2.46 (1 H, d), 2.26-2.40 (1 H, m), 1.88-2.16 (3 H, m). LCMS RT = 1.90; [M] ⁺ = 370.28.

Example 258
(R) -N- (5-Methyl-1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ＤＭＦ(１０ml)中の５−メチル−１,３,４−オキサジアゾール−２−アミン(１.９２ｇ, ２０mmol)の溶液に、ＮａＯＨ(２０Ｍ, ２ml)、ＣＳ_２(３ml)、ＮａＯＨ(２０Ｍ, ２ml)およびヨードメタン(３ml)を、ゆっくりと１０分かけて加えた。混合物を室温で１時間撹拌し、２０mlの水に注いだ。沈殿した固体を濾過し、水で洗浄し、望ましい生成物を乾燥させ、５−メチル−１,３,４−オキサジアゾール−２−イルカルボンイミドジチオ酸ジメチルを白色の固体として得た(１.４５ｇ, ３５.７％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 2.63 (s, 6H), 2.50 (s, 3H). LCMS R.T. 1.66分; [M+H] = 203.91。 Step A: Dimethyl 5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate

To a solution of 5-methyl-1,3,4-oxadiazol-2-amine (1.92 g, 20 mmol) in DMF (10 ml) was added NaOH (20 M, 2 ml), CS ₂ (3 ml), NaOH (20 M , 2 ml) and iodomethane (3 ml) were slowly added over 10 minutes. The mixture was stirred at room temperature for 1 hour and poured into 20 ml water. The precipitated solid was filtered, washed with water and the desired product was dried to give dimethyl 5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate as a white solid (1 .45 g, 35.7%).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 2.63 (s, 6H), 2.50 (s, 3H). LCMS RT 1.66 min; [M + H] = 203.91.

ＤＭＦ(５ml)中の、５−メチル−１,３,４−オキサジアゾール−２−イルカルボンイミドジチオ酸ジメチル(２６０mg, １.２８mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２００mg, １.２８mmol)および炭酸セシウム(８７６mg, ２.６９mmol)の混合物を、室温で一夜撹拌した。混合物を濃縮し、Biotageシリカゲルカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−(５−メチル−１,３,４−オキサジアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(１９２mg, ５４.１％)。
¹H NMR (500 MHz, MeOD) δ 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分。 Step B: (R) -N- (5-Methyl-1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

Dimethyl 5-methyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate (260 mg, 1.28 mmol), (S) -3- (aminomethyl) quinuclidine-3 in DMF (5 ml) A mixture of -ol dihydrochloride (200 mg, 1.28 mmol) and cesium carbonate (876 mg, 2.69 mmol) was stirred overnight at room temperature. The mixture is concentrated and purified on a Biotage silica gel column (100% ethyl acetate, then 10-35% 9: 1 methanol: ammonium hydroxide-chloroform), which is the desired product (R) -N- (5-methyl -1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine (192 mg, 54.1%).
¹ H NMR (500 MHz, MeOD) δ 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. RT 0.16 min.

Example 259
(R) -N- (3-Methyl-1,2,4-thiadiazol-5-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

ＤＭＦ(１０ml)中の、３−メチル−１,２,４−チアジアゾール−５−アミン(２.３ｇ, ２０mmol)の溶液に、ＮａＯＨ(２０Ｍ, ２ml)、ＣＳ_２(３ml)、ＮａＯＨ(２０Ｍ, ２ml)およびヨードメタン(３ml)を１０分かけてゆっくりと加えた。混合物を室温で１時間撹拌し、２０mlの水に注いだ。沈殿した固体を濾過し、水で洗浄し、乾燥させ、純粋でない３−メチル−１,２,４−チアジアゾール−５−イルカルボンイミドジチオ酸ジメチルを黄色の固体として得た(２.３ｇ, ５２.５％)。
¹H NMR (500 MHz, CDCl3) δ ppm 2.67 (s), 2.62 (s). MS [M+H] = 219.85。 Step A: Dimethyl 3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate

To a solution of 3-methyl-1,2,4-thiadiazol-5-amine (2.3 g, 20 mmol) in DMF (10 ml) was added NaOH (20 M, 2 ml), CS ₂ (3 ml), NaOH (20 M, 2 ml) and iodomethane (3 ml) were added slowly over 10 minutes. The mixture was stirred at room temperature for 1 hour and poured into 20 ml water. The precipitated solid was filtered, washed with water and dried to give impure dimethyl 3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate as a yellow solid (2.3 g, 52 .5%).
< ¹ > H NMR (500 MHz, CDCl3) [delta] ppm 2.67 (s), 2.62 (s). MS [M + H] = 219.85.

ＤＭＦ(５ml)中の、３−メチル−１,２,４−チアジアゾール−５−イルカルボンイミドジチオ酸ジメチル(２８１mg, １.２８mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２００mg, １.２８mmol)および炭酸セシウム(８７６mg, ２.６９mmol)の混合物を、室温で一夜撹拌した。混合物を濃縮し、Biotageシリカゲルカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−(３−メチル−１,２,４−チアジアゾール−５−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(１４７.８mg, ４０.５％)。
¹H NMR (500 MHz, MeOD) δ 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分。 Step B: (R) -N- (3-Methyl-1,2,4-thiadiazol-5-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 3-methyl-1,2,4-thiadiazol-5-ylcarbonimidodithioate (281 mg, 1.28 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol in DMF (5 ml) A mixture of dihydrochloride (200 mg, 1.28 mmol) and cesium carbonate (876 mg, 2.69 mmol) was stirred at room temperature overnight. The mixture was concentrated and purified on a Biotage silica gel column (100% ethyl acetate, then 10-35% 9: 1 methanol: ammonium hydroxide-chloroform), the desired product (R) -N- (3-methyl -1,2,4-thiadiazol-5-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine (147.8 mg, 40.5%).
¹ H NMR (500 MHz, MeOD) δ 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; RT = 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] octane] -2-amine

ＤＭＦ(５ml)中の３−メチル−１,２,４−オキサジアゾール−５−アミン(４９０mg, ４.９４mmol)の溶液に、ＮａＯＨ(２０Ｍ, ０.５ml)、ＣＳ_２(１ml)、ＮａＯＨ(２０Ｍ, ０.５ml)およびヨードメタン(１ml)を、１０分かけてゆっくりと加えた。混合物を室温で１時間撹拌した。混合物は非常に濃厚となった。２０mlの水を加えた。固体を濾過して取り、水で洗浄し、乾燥させ、純粋でない３−メチル−１,２,４−オキサジアゾール−５−イルカルボンイミドジチオ酸ジメチルである黄色の固体を得た(７７０mg, ７７％)。
MS (LCMS) [M+H] = 203.91; R.T. = 1.84分。
生成物を次の工程に直接用いた。 Step A: Dimethyl 3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate

To a solution of 3-methyl-1,2,4-oxadiazol-5-amine (490 mg, 4.94 mmol) in DMF (5 ml) was added NaOH (20 M, 0.5 ml), CS ₂ (1 ml), NaOH. (20M, 0.5 ml) and iodomethane (1 ml) were added slowly over 10 minutes. The mixture was stirred at room temperature for 1 hour. The mixture became very thick. 20 ml of water was added. The solid was filtered off, washed with water and dried to give a yellow solid which was impure dimethyl 3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate (770 mg, 77%).
MS (LCMS) [M + H] = 203.91; RT = 1.84 min.
The product was used directly in the next step.

ＤＭＦ(５ml)中の３−メチル−１,２,４−オキサジアゾール−５−イルカルボンイミドジチオ酸ジメチル(２８０mg, １.３７mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２１５mg, １.３７mmol)および炭酸セシウム(９４２mg, ２.８９mmol)の混合物を、室温で一夜撹拌した。混合物を濃縮し、Biotageシリカゲルカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−(３−メチル−１,２,４−オキサジアゾール−５−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(１９８mg, ５１.９％)。
¹H NMR (500 MHz, MeOD) δ 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分。 Step B: (R) -N- (3-Methyl-1,2,4-oxadiazol-5-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2 ] Octane] -2-amine

Dimethyl 3-methyl-1,2,4-oxadiazol-5-ylcarbonimidodithioate (280 mg, 1.37 mmol), (S) -3- (aminomethyl) quinuclidine-3- in DMF (5 ml) A mixture of all dihydrochloride (215 mg, 1.37 mmol) and cesium carbonate (942 mg, 2.89 mmol) was stirred at room temperature overnight. The mixture was concentrated and purified on a Biotage silica gel column (100% ethyl acetate, then 10-35% 9: 1 methanol: ammonium hydroxide-chloroform), the desired product (R) -N- (3-methyl -1,2,4-oxadiazol-5-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine (198 mg, 51.9%).
¹ H NMR (500 MHz, MeOD) δ 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; RT = 0.26 min.

Example 261
(R) -N- (6- (Methylthio) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

Ｎ,Ｎ−ジメチルホルムアミド(１２ml)中の６−クロロピリミジン−４−アミン(１.２９５ｇ, １０mmol)の溶液に、ＮａＯＨ(１ml, ２０.００mmol, ２０Ｍ)、ＣＳ_２(１.５ml, ２４.８８mmol)、ＮａＯＨ(１ml, ２０.００mmol, ２０Ｍ)およびヨードメタン(１.５ml, ２３.９９mmol)を、１５分間で滴下した。撹拌を１.５時間続けて、混合物を水に注いだ。橙色の固体を分離し、水で洗浄し、乾燥させ、メタノールから再結晶し、６−クロロピリミジン−４−イルカルボンイミドジチオ酸ジメチルを黄色の固体として得た(０.９６６ｇ, ４.１３mmol, 収率４１.３％)。
LCMS R.T. = 2.39; [M+H]⁺ = 234.08。 Step A: Dimethyl 6-chloropyrimidin-4-ylcarbonimidodithioate

To a solution of 6-chloropyrimidin-4-amine (1.295 g, 10 mmol) in N, N-dimethylformamide (12 ml) was added NaOH (1 ml, 20.00 mmol, 20M), CS ₂ (1.5 ml, 24. 88 mmol), NaOH (1 ml, 20.00 mmol, 20 M) and iodomethane (1.5 ml, 23.99 mmol) were added dropwise over 15 minutes. Stirring was continued for 1.5 hours and the mixture was poured into water. The orange solid was separated, washed with water, dried and recrystallized from methanol to give dimethyl 6-chloropyrimidin-4-ylcarbonimidodithioate as a yellow solid (0.966 g, 4.13 mmol, Yield 41.3%).
LCMS RT = 2.39; [M + H] ⁺ = 234.08.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.９ｇ, ３.９３mmol)に、Ｃｓ_２ＣＯ_３(２.６９ｇ, ８.２５mmol)および６−クロロピリミジン−４−イルカルボンイミドジチオ酸ジメチル(０.９６４ｇ, ４.１２mmol)を加えた。懸濁液を室温で１８時間撹拌し、１００℃で３時間加熱した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜１５％ ９：１ メタノール：水酸化アンモニウム−酢酸エチル)、(Ｒ)−Ｎ−(６−(メチルチオ)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを薄黄色の固体として得た(０.２１ｇ, ０.７２mmol, 収率４８.２％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.43 (1 H, br. s.), 8.55 (1 H, d), 6.78 (1 H, br. s.), 3.98 (1 H, d), 3.64 (1 H, d), 3.37 (1 H, dd), 2.72 - 3.06 (5 H, m), 2.51 (3 H, s), 2.08 - 2.24 (2 H, m), 1.69 - 1.81 (1 H, m), 1.41 - 1.64 (2 H, m). LCMS R.T. = 0.93; [M+H]⁺ = 306.29。 Step B: (R) -N- (6- (Methylthio) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.9 g, 3.93 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (2.69 g, 8.25 mmol) and dimethyl 6-chloropyrimidin-4-ylcarbonimidodithioate (0.964 g, 4.12 mmol) were added. The suspension was stirred at room temperature for 18 hours and heated at 100 ° C. for 3 hours. The mixture was concentrated and purified by chromatography on silica gel (5-15% 9: 1 methanol: ammonium hydroxide-ethyl acetate), (R) -N- (6- (methylthio) pyrimidin-4-yl) -4H -1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a pale yellow solid (0.21 g, 0.72 mmol, 48.2% yield). ).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.43 (1 H, br.s.), 8.55 (1 H, d), 6.78 (1 H, br.s.), 3.98 (1 H, d), 3.64 (1 H, d), 3.37 (1 H, dd), 2.72-3.06 (5 H, m), 2.51 (3 H, s), 2.08-2.24 (2 H, m), 1.69-1.81 (1 H , m), 1.41-1.64 (2 H, m). LCMS RT = 0.93; [M + H] ⁺ = 306.29.

Example 262
(R) -N-([1,2,4] triazolo [4,3-a] pyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

ジクロロメタン(１Ｌ)中の１Ｈ−イミダゾール(４２ｇ, ６１７mmol)の溶液に、臭化シアン(２２.５, ２１２mmol)を加え、混合物を３０分間還流し、室温まで冷却し、白色の固体を濾過して取った。濾液を１００mlまで濃縮し、冷蔵庫で３日間保存した。沈殿した固体を濾過して取り、８ｇのジ(１Ｈ−イミダゾール−１−イル)メタンイミンを得た(４９.６mmol, ８％)。
¹H NMR (500 MHz, DMSO) δ ppm 8.09 (s, 1H), 7.55 (s, 1H), 7.13 (s, 1H)。 Step A: Di (1H-imidazol-1-yl) methanimine

To a solution of 1H-imidazole (42 g, 617 mmol) in dichloromethane (1 L) was added cyanogen bromide (22.5, 212 mmol), the mixture was refluxed for 30 min, cooled to room temperature, and the white solid was filtered. I took it. The filtrate was concentrated to 100 ml and stored in the refrigerator for 3 days. The precipitated solid was filtered off to give 8 g of di (1H-imidazol-1-yl) methanimine (49.6 mmol, 8%).
¹ H NMR (500 MHz, DMSO) δ ppm 8.09 (s, 1H), 7.55 (s, 1H), 7.13 (s, 1H).

ＴＨＦ(７０ml)中の２−ヒドラジニルピリジン(５.２ｇ, ４７.６mmol)の溶液に、ジ(１Ｈ−イミダゾール−１−イル)メタンイミン(７.８ｇ, ４８.４mmol)を加えた。混合物を一夜還流した。粗製の混合物を蒸発させ、Biotageシリカゲルカラムで精製し(０〜２５％, メタノール−塩化メチレン)、紫色のスポットを集め、[１,２,４]トリアゾロ[４,３−ａ]ピリジン−３−アミンを得た(４.７ｇ, ３５mmol, ７３.５％)。
¹H NMR (500 MHz, DMSO) δ 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分。 Step B: [1,2,4] Triazolo [4,3-a] pyridin-3-amine

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 refluxed overnight. The crude mixture was evaporated and purified on a Biotage silica gel column (0-25%, methanol-methylene chloride) and the purple spot was collected and [1,2,4] triazolo [4,3-a] pyridine-3- The amine was obtained (4.7 g, 35 mmol, 73.5%).
¹ H NMR (500 MHz, DMSO) δ 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 MS (LCMS) [M + H] = 134.98; RT = 0.1 min.

ＤＭＦ(５ml)中の[１,２,４]トリアゾロ[４,３−ａ]ピリジン−３−アミン(３００mg, ２.２４mmol)の溶液に、ＮａＯＨ(２０Ｍ, ０.２５ml)、ＣＳ_２(０.５ml)、ＮａＯＨ(２０Ｍ, ０.２５ml)およびヨードメタン(０.５ml)を１０分かけてゆっくりと加えた。混合物を室温で１時間撹拌し、１０mlの水を反応混合物に加えた。沈殿した固体を濾過して取り、水(１００ml)で洗浄し、乾燥させ、２３０mgの[１,２,４]トリアゾロ[４,３−ａ]ピリジン−３−イルカルボンイミドジチオ酸ジメチルを白色の固体として得た(０.９６mmol, ４３.１％)。
¹H NMR (500 MHz, CDCl3) δ 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分。 Step C: [1,2,4] Triazolo [4,3-a] pyridin-3-ylcarbonimidodithioate dimethyl

To a solution of [1,2,4] triazolo [4,3-a] pyridin-3-amine (300 mg, 2.24 mmol) in DMF (5 ml) was added NaOH (20 M, 0.25 ml), CS ₂ (0 0.5 ml), NaOH (20M, 0.25 ml) and iodomethane (0.5 ml) were added slowly over 10 minutes. The mixture was stirred at room temperature for 1 hour and 10 ml of water was added to the reaction mixture. The precipitated solid was filtered off, washed with water (100 ml), dried and 230 mg of [1,2,4] triazolo [4,3-a] pyridin-3-ylcarbonimidodithioate dimethyl was added to a white solution. Obtained as a solid (0.96 mmol, 43.1%).
¹ H NMR (500 MHz, CDCl3) δ 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; RT = 1.26 min.

ＤＭＦ(５ml)中の、[１,２,４]トリアゾロ[４,３−ａ]ピリジン−３−イルカルボンイミドジチオ酸ジメチル(１２０mg, ０.５０mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１２０mg, ０.７６mmol)および炭酸セシウム(４９２mg, １.５mmol)の混合物を、７０℃で６時間加熱した。混合物を濃縮し、Biotageシリカゲルカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−([１,２,４]トリアゾロ[４,３−ａ]ピリジン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(９７.２mg, ６１.５％)。
¹H NMR (500 MHz, CDCl₃) δ 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分。 Step D: (R) -N-([1,2,4] triazolo [4,3-a] pyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2. 2.2] Octane] -2-amine

Dimethyl [1,2,4] triazolo [4,3-a] pyridin-3-ylcarbonimidodithioate (120 mg, 0.50 mmol), (S) -3- (aminomethyl) in DMF (5 ml) A mixture of quinuclidin-3-ol dihydrochloride (120 mg, 0.76 mmol) and cesium carbonate (492 mg, 1.5 mmol) was heated at 70 ° 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), the desired product (R) -N-([1, 2,4] Triazolo [4,3-a] pyridin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained. (97.2 mg, 61.5%).
¹ H NMR (500 MHz, CDCl ₃ ) δ 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; RT = 1.22 min.

Example 263
(R) -N- (6-bromothiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

ＤＭＦ(３ml)中の６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−アミン(７００mg, ３.０３mmol)の懸濁液に、１６.０Ｍ 水酸化ナトリウム(４００μｌ, ６.４０mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で二硫化炭素(４５０μｌ, ７.５７mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに１６.０Ｍ 水酸化ナトリウム(４００μｌ, ６.４０mmol)を加え、混合物を再度１０分間撹拌した。最後に、ヨードメタン(４５０μｌ, ７.２７mmol)を滴下した。混合物を５分間撹拌し、この時点で多量の黄色の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、さらに精製することなく用いるのに十分な純度の黄色の固体として６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−イルカルボンイミドジチオ酸ジメチルを得た(６８０mg, 収率６７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.63 (s, 1 H) 2.68 (s, 6 H)。 Step A: Dimethyl 6-bromothiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate

To a suspension of 6-bromothiazolo [5,4-b] pyrazin-2-amine (700 mg, 3.03 mmol) in DMF (3 ml) was added 16.0 M sodium hydroxide (400 μl, 6.40 mmol). . The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (450 μl, 7.57 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. Further 16.0M sodium hydroxide (400 μl, 6.40 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (450 μl, 7.27 mmol) was added dropwise. The mixture was stirred for 5 minutes, at which point a large amount of yellow precipitate had formed. The mixture is poured into water and the solid is collected by filtration to give dimethyl 6-bromothiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate as a yellow solid of sufficient purity to be used without further purification. (680 mg, 67% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.63 (s, 1 H) 2.68 (s, 6 H).

アセトニトリル(２５ml)中の、６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−イルカルボンイミドジチオ酸ジメチル(３００mg, ０.８９５mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２１０mg, ０.８９５mmol)および炭酸セシウム(６００mg, １.７９mmol)の混合物を、開放フラスコ中で、メタンチオールの除去を助けるために全反応時間窒素を溶液に通気しながら、１００℃の油浴で２時間加熱した。２時間後、ＴＬＣにより反応が完了したことが示され、混合物を環境温度まで冷却し、水で希釈し、真空で濃縮した。混合物をクロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、粗製の残渣をシリカゲルのクロマトグラフィーによって精製した(２〜４０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)。生成物のフラクションを合わせ、真空で濃縮し、(Ｒ)−Ｎ−(６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２００mg, 収率５７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.39 (br. s., 1 H) 8.48 (s, 1 H) 4.05 (d, J=9.79 Hz, 1 H) 3.72 (d, J=9.79 Hz, 1 H) 3.42 (dd, J=15.06, 1.76 Hz, 1 H) 2.73 - 3.08 (m, 5 H) 2.10 - 2.22 (m, 2 H) 1.73 - 1.84 (m, J=14.09, 9.94, 4.17, 4.17 Hz, 1 H) 1.52 - 1.65 (m, 2 H). MS (LC/MS) R.T. = 1.29; [M+H]⁺ = 394.99。 Step B: (R) -N- (6-Bromothiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 6-bromothiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate (300 mg, 0.895 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol in acetonitrile (25 ml) A mixture of dihydrochloride (210 mg, 0.895 mmol) and cesium carbonate (600 mg, 1.79 mmol) was placed in an open flask at 100 ° C. while bubbling nitrogen through the solution for a total reaction time to help remove methanethiol. In an oil bath for 2 hours. After 2 hours, TLC showed that the reaction was complete and the mixture was cooled to ambient temperature, diluted with water and concentrated in vacuo. The mixture was extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered, concentrated in vacuo, and the crude residue was purified by chromatography on silica gel (2-40% 9: 1 methanol: ammonium hydroxide-chloroform). . The product fractions were combined, concentrated in vacuo, and (R) -N- (6-bromothiazolo [5,4-b] pyrazin-2-yl) -4H-1'-azaspiro [oxazole-5,3'- Bicyclo [2.2.2] octane] -2-amine was obtained (200 mg, 57% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.39 (br. S., 1 H) 8.48 (s, 1 H) 4.05 (d, J = 9.79 Hz, 1 H) 3.72 (d, J = 9.79 Hz, 1 H) 3.42 (dd, J = 15.06, 1.76 Hz, 1 H) 2.73-3.08 (m, 5 H) 2.10-2.22 (m, 2 H) 1.73-1.84 (m, J = 14.09, 9.94, 4.17, 4.17 Hz, 1 H) 1.52-1.65 (m, 2 H). MS (LC / MS) RT = 1.29; [M + H] ⁺ = 394.99.

ＤＭＦ(１.５ml)中の、６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−イルカルボンイミドジチオ酸ジメチル(実施例２６３の工程Ａより)(１００mg, ０.２９８mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(６８mg, ０.２９８mmol)および炭酸セシウム(１００mg, ０.６０mmol)の混合物を、１ドラムのバイアルに入れ、１００℃の油浴で１時間加熱し、この時点で、ナトリウム チオメトキシド(１００mg, １.４３mmol)を加え、混合物を一夜加熱した。混合物を環境温度まで冷却し、水(２０ml)に注ぎ、得られた固体を濾過によって集め、シリカゲルのクロマトグラフィーによって精製した(２〜４０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)。生成物のフラクションを合わせて真空で濃縮し、(Ｒ)−Ｎ−(６−(メチルチオ)チアゾロ[５,４−ｂ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(５２mg, 収率４６％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.39 (br. s., 1 H) 8.31 (s, 1 H) 4.03 (d, J=9.77 Hz, 1 H) 3.70 (d, J=9.77 Hz, 1 H) 3.41 (dd, J=14.95, 1.83 Hz, 1 H) 2.73 - 3.10 (m, 5 H) 2.63 (s, 3 H) 2.10 - 2.25 (m, 2 H) 1.47 - 1.86 (m, 3 H). MS (LC/MS) R.T. = 1.04; [M+H]⁺ = 363.04。 Example 264
(R) -N- (6- (Methylthio) thiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 6-bromothiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate (from Step A of Example 263) (100 mg, 0.298 mmol), (S)-in DMF (1.5 ml) A mixture of 3- (aminomethyl) quinuclidin-3-ol dihydrochloride (68 mg, 0.298 mmol) and cesium carbonate (100 mg, 0.60 mmol) was placed in a 1-dram vial and placed in a 100 ° C. oil bath for 1 hour. At this point, sodium thiomethoxide (100 mg, 1.43 mmol) was added and the mixture was heated overnight. The mixture was cooled to ambient temperature, poured into water (20 ml) and the resulting solid was collected by filtration and purified by chromatography on silica gel (2-40% 9: 1 methanol: ammonium hydroxide-chloroform). The product fractions were combined and concentrated in vacuo to give (R) -N- (6- (methylthio) thiazolo [5,4-b] pyrazin-2-yl) -4H-1'-azaspiro [oxazole-5, 3'-bicyclo [2.2.2] octane] -2-amine was obtained (52 mg, 46% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.39 (br. S., 1 H) 8.31 (s, 1 H) 4.03 (d, J = 9.77 Hz, 1 H) 3.70 (d, J = 9.77 Hz, 1 H) 3.41 (dd, J = 14.95, 1.83 Hz, 1 H) 2.73-3.10 (m, 5 H) 2.63 (s, 3 H) 2.10-2.25 (m, 2 H) 1.47-1.86 (m, 3 H ). MS (LC / MS) RT = 1.04; [M + H] ⁺ = 363.04.

Example 265
(R) -N- (5-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

５−クロロチアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチル(２５０mg, ０.９６６mmol)をＭｅＯＨ(１０ml)に懸濁し、メタノール中２５％(w/w) ナトリウム メトキシドの溶液(１０ml, ４６.３mmol)を加えた。得られた溶液を一夜還流し、環境温度まで冷却し、等量の水に注ぎ、クロロホルム(４×)で抽出した。水相に大量の化合物が残っており、そのためこれを濃縮して残渣を得て、少量の１Ｎ ＨＣｌ(得られた溶液を酸性にする程ではない)に溶解し、ＥｔＯＡｃ(５×)で再度抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。５−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−アミンを白色の固体として得た(１４４mg, ０.７９０mmol, 収率８２％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.43 (s, 1 H) 7.81 (s, 2 H) 3.90 (s, 3 H). MS (LC/MS) R.T. = 0.73; [M+H]⁺ = 183.03。 Step A: 5-Methoxythiazolo [5,4-d] pyrimidin-2-amine

Ethyl 5-chlorothiazolo [5,4-d] pyrimidin-2-ylcarbamate (250 mg, 0.966 mmol) was suspended in MeOH (10 ml) and a solution of 25% (w / w) sodium methoxide in methanol (10 ml, 46.3 mmol) was added. The resulting solution was refluxed overnight, cooled to ambient temperature, poured into an equal volume of water and extracted with chloroform (4 ×). A large amount of compound remains in the aqueous phase, so it is concentrated to give a residue, dissolved in a small amount of 1N HCl (not enough to acidify the resulting solution) and again with EtOAc (5 ×). Extracted. The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. 5-Methoxythiazolo [5,4-d] pyrimidin-2-amine was obtained as a white solid (144 mg, 0.790 mmol, 82% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.43 (s, 1 H) 7.81 (s, 2 H) 3.90 (s, 3 H). MS (LC / MS) RT = 0.73; (M + H ] ⁺ = 183.03.

ＤＭＦ(５ml)中の５−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−アミン(９１１mg, ５.００mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(５００μｌ, １０.００mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で、二硫化炭素(７５０μｌ, １２.５０mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(５００μｌ, １０.００mmol)を加え、混合物を再度１０分間撹拌した。最後に、ヨードメタン(７５０μｌ, １２.００mmol)を滴下した。混合物を５分間撹拌し、この時点で、多量の黄色の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、黄色の固体を得た。これをさらにシリカゲルのクロマトグラフィーによって精製し(２〜２０％ ＥｔＯＡｃ／ＣＨＣｌ_３)、５−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチルを得た(３８０mg, 収率２７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.90 (s, 1 H) 4.09 (s, 3 H) 2.66 (s, 6 H)。 Step B: Dimethyl 5-methoxythiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate

To a suspension of 5-methoxythiazolo [5,4-d] pyrimidin-2-amine (911 mg, 5.00 mmol) in DMF (5 ml) was added 20.0 M sodium hydroxide (500 μl, 10.00 mmol). added. The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (750 μl, 12.50 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (500 μl, 10.00 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (750 μl, 12.00 mmol) was added dropwise. The mixture was stirred for 5 minutes, at which point a large amount of yellow precipitate had formed. The mixture was poured into water and the solid was collected by filtration to give a yellow solid. This was further purified by chromatography on silica gel (2-20% EtOAc / CHCl ₃ ) to give dimethyl 5-methoxythiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (380 mg, yield). Rate 27%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.90 (s, 1 H) 4.09 (s, 3 H) 2.66 (s, 6 H).

ＤＭＦ(１.７ml)中の、５−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチル(１００mg, ０.３４９mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(８０mg, ０.３４９mmol)および炭酸セシウム(２２８mg, ０.６９８mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、固体を濾過によって集め、(Ｒ)−Ｎ−(５−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(７８mg, 収率６４％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.12 (br. s., 1 H) 8.63 (s, 1 H) 3.95 - 4.18 (m, 4 H) 3.71 (d, J=9.77 Hz, 1 H) 3.41 (d, J=15.26 Hz, 1 H) 2.74 - 3.10 (m, 5 H) 2.11 - 2.27 (m, 2 H) 1.71 - 1.86 (m, 1 H) 1.50 - 1.70 (m, 2 H). MS (LC/MS) R.T. = 1.66; [M+H]⁺ = 347.0。 Step C: (R) -N- (5-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

Dimethyl 5-methoxythiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (100 mg, 0.349 mmol), (S) -3- (aminomethyl) quinuclidine in DMF (1.7 ml). A mixture of -3-ol dihydrochloride (80 mg, 0.349 mmol) and cesium carbonate (228 mg, 0.698 mmol) was heated at 100 ° C. for 2 hours. The reaction mixture is cooled to ambient temperature, poured into water, the solid is collected by filtration, and (R) -N- (5-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1′- Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (78 mg, 64% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.12 (br. S., 1 H) 8.63 (s, 1 H) 3.95-4.18 (m, 4 H) 3.71 (d, J = 9.77 Hz, 1 H) 3.41 (d, J = 15.26 Hz, 1 H) 2.74-3.10 (m, 5 H) 2.11-2.27 (m, 2 H) 1.71-1.86 (m, 1 H) 1.50-1.70 (m, 2 H). MS (LC / MS) RT = 1.66; [M + H] ⁺ = 347.0.

Example 266
(R) -N- (5-Ethyl-1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ＤＭＦ(１０ml)中の５−エチル−１,３,４−オキサジアゾール−２−アミン(２.２６ｇ, ２０mmol)の溶液に、ＮａＯＨ(２０Ｍ, ２ml)、ＣＳ_２(３ml)、ＮａＯＨ(２０Ｍ, ２ml)およびヨードメタン(３ml)を１０分かけてゆっくりと加えた。混合物を室温で２時間撹拌し、３０mlの水に注いだ。沈殿した黄色の固体を濾過して取り、水で洗浄し、乾燥させ、望ましい生成物である５−エチル−１,３,４−オキサジアゾール−２−イルカルボンイミドジチオ酸ジメチルを白色の固体として得た(２.６ｇ, ５９.８％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 2.86-2.83 (q, 2H), 2.63 (s, 6H), 1.39-1.35 (t, 3H). MS (LCMS) [M+H] = 217.95; R.T. = 1.93分。 Step A: Dimethyl 5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate

To a solution of 5-ethyl-1,3,4-oxadiazol-2-amine (2.26 g, 20 mmol) in DMF (10 ml) was added NaOH (20 M, 2 ml), CS ₂ (3 ml), NaOH (20 M , 2 ml) and iodomethane (3 ml) were added slowly over 10 minutes. The mixture was stirred at room temperature for 2 hours and poured into 30 ml water. The precipitated yellow solid is filtered off, washed with water, dried and the desired product, dimethyl 5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate, is a white solid (2.6 g, 59.8%).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 2.86-2.83 (q, 2H), 2.63 (s, 6H), 1.39-1.35 (t, 3H). MS (LCMS) [M + H] = 217.95; RT = 1.93 minutes.

ＤＭＦ(１０ml)中の、５−エチル−１,３,４−オキサジアゾール−２−イルカルボンイミドジチオ酸ジメチル(３２７mg, １.５mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２３５mg, １.５mmol)および炭酸セシウム(１０００mg, ３.１６mmol)の混合物を、室温で一夜撹拌した。混合物を濃縮し、Biotageシリカゲルのカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−(５−エチル−１,３,４−オキサジアゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２９０mg, ６６％)。
¹H NMR (500 MHz, MeOD) δ 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分。 Step B: (R) -N- (5-Ethyl-1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

Dimethyl 5-ethyl-1,3,4-oxadiazol-2-ylcarbonimidodithioate (327 mg, 1.5 mmol), (S) -3- (aminomethyl) quinuclidine-3 in DMF (10 ml) A mixture of -ol dihydrochloride (235 mg, 1.5 mmol) and cesium carbonate (1000 mg, 3.16 mmol) was stirred overnight at room temperature. The mixture was concentrated and purified on a column of Biotage silica gel (100% ethyl acetate, then 10-35% 9: 1 methanol: ammonium hydroxide-chloroform), the desired product (R) -N- (5- Ethyl-1,3,4-oxadiazol-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (290 mg). , 66%).
¹ H NMR (500 MHz, MeOD) δ 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). MS (LCMS) [M + H] = 278.09; RT = 0.48 min.

Example 267
(R) -N- (3,5-dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

ジクロロメタン(２５ml)中の３,５−ジクロロピリジン−２−アミン(０.３６ｇ, ２.２０９mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.５２３ｇ, ２.２５３mmol)を加えた。反応物を４０℃で３時間撹拌した。反応物を室温まで冷却し、粗製の物質をクロマトグラフィーによって精製し(Biotage, ２５〜１００％ 酢酸エチル／ヘキサン)、３,５−ジクロロ−２−イソチオシアナトピリジンを得た(０.４ｇ, １.９５１mmol, 収率８８％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.50 (t, J=2.59 Hz, 1 H), 8.45 (t, J=2.59 Hz, 1 H). MS (LC/MS) R.T. = 2.07; [M+H]⁺ = 204.8。 Step A: 3,5-dichloro-2-isothiocyanatopyridine

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) was added. The reaction was stirred at 40 ° C. for 3 hours. The reaction was cooled to room temperature and the crude material was purified by chromatography (Biotage, 25-100% ethyl acetate / hexanes) to give 3,5-dichloro-2-isothiocyanatopyridine (0.4 g, 1.951 mmol, 88% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.50 (t, J = 2.59 Hz, 1 H), 8.45 (t, J = 2.59 Hz, 1 H). MS (LC / MS) RT = 2.07; [ M + H] ⁺ = 204.8.

Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中の３,５−ジクロロ−２−イソチオシアナトピリジン(０.１１ｇ, ０.５５mmol)に、Ｅｔ_３Ｎ(０.１７ml, １.２１mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１３ｇ, ０.５６mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＮ,Ｎ−ジメチルホルムアミド(１０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.２６ml, １.６５mmol)で処理した。反応物を７０℃で２時間加熱した。反応物を室温まで冷却し、真空で濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(３,５−ジクロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０８ｇ, ０.２４mmol, 収率４４％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.91 (s, 1 H), 8.11 - 8.17 (m, 1 H), 7.97 (d, J=2.44 Hz, 1 H), 3.84 (d, J=9.77 Hz, 1 H), 3.59 (d, J=9.77 Hz, 1 H), 2.95 - 3.04 (m, 2 H), 2.72 - 2.81 (m, 2 H), 2.66 (t, J=7.63 Hz, 2 H), 2.01 (s, 1 H), 1.89 (s, 1 H), 1.54 - 1.62 (m, 2 H), 1.42 - 1.50 (m, 1 H). MS (LC/MS) R.T. = 0.78; [M+]⁺ = 326.1。 Step B: (R) -N- (3,5-Dichloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

3,5-Dichloro-2-isothiocyanatopyridine (0.11 g, 0.55 mmol) in N, N-dimethylformamide (10 ml) was added to Et ₃ N (0.17 ml, 1.21 mmol) and (S). -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.13 g, 0.56 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with N, N-dimethylformamide (10 ml) and N, N′-diisopropylcarbodiimide (0.26 ml, 1.65 mmol). The reaction was heated at 70 ° C. for 2 hours. The reaction was cooled to room temperature and concentrated in vacuo to give the crude product. The crude product was purified by chromatography _{(Biotage, 85% CHCl 3,} 14% MeOH, 1% NH 4 OH), (R) -N- (3,5- dichloro-pyridin-2-yl)-4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.08 g, 0.24 mmol, 44% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.91 (s, 1 H), 8.11-8.17 (m, 1 H), 7.97 (d, J = 2.44 Hz, 1 H), 3.84 (d, J = 9.77 Hz, 1 H), 3.59 (d, J = 9.77 Hz, 1 H), 2.95-3.04 (m, 2 H), 2.72-2.81 (m, 2 H), 2.66 (t, J = 7.63 Hz, 2 H), 2.01 (s, 1 H), 1.89 (s, 1 H), 1.54-1.62 (m, 2 H), 1.42-1.50 (m, 1 H). MS (LC / MS) RT = 0.78; [ M +] ⁺ = 326.1.

Example 268
(R) -N- (5-chlorothiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

ＤＭＦ(５ml)中の５−クロロチアゾロ[５,４−ｂ]ピリジン−２−アミン(９３０mg, ５.００mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(５００μｌ, １０.００mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で、二硫化炭素(７５０μｌ, １２.５０mmol)を加え、混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(５００μｌ, １０.０mmol)を加え、混合物を再度１０分間撹拌した。最後に、ヨードメタン(７５０μｌ, １２.００mmol)を滴下した。添加中は発熱に注意した。混合物を１５分間撹拌し、この時点で、多量の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集めた。集めた固体の大部分が薄黄色であり、結晶性であった。少量の塊の僅かに暗い橙色のゴム状固体が存在し、これらを手で除いて廃棄した。残ったものが表題化合物５−クロロチアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルであった(１.００ｇ, 収率６９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.04 (d, J=8.53 Hz, 1 H) 7.38 (d, J=8.53 Hz, 1 H) 2.66 (s, 6 H)。 Step A: Dimethyl 5-chlorothiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate

To a suspension of 5-chlorothiazolo [5,4-b] pyridin-2-amine (930 mg, 5.00 mmol) in DMF (5 ml) was added 20.0 M sodium hydroxide (500 μl, 10.00 mmol). . The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (750 μl, 12.50 mmol) was added and the mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (500 μl, 10.0 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (750 μl, 12.00 mmol) was added dropwise. Care was taken to avoid exotherm during the addition. The mixture was stirred for 15 minutes, at which point a large amount of precipitate had formed. The mixture was poured into water and the solid was collected by filtration. Most of the collected solid was light yellow and crystalline. There were small chunks of slightly dark orange rubbery solids that were removed by hand and discarded. What remained was the title compound dimethyl 5-chlorothiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate (1.00 g, 69% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.04 (d, J = 8.53 Hz, 1 H) 7.38 (d, J = 8.53 Hz, 1 H) 2.66 (s, 6 H).

ＤＭＦ(１.７ml)中の、５−クロロチアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(１００mg, ０.３５mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(７９mg, ０.３５mmol)および炭酸セシウム(２２５mg, ０.６９mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、固体を濾過によって集めた。粗製の固体をシリカゲルのクロマトグラフィーによって精製し(２〜４０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、(Ｒ)−Ｎ−(５−クロロチアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(６２mg, 収率５１％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.13 (br. s., 1 H) 7.93 (d, J=8.53 Hz, 1 H) 7.45 (d, J=8.28 Hz, 1 H) 3.92 (d, J=10.29 Hz, 1 H) 3.67 (d, J=10.29 Hz, 1 H) 3.00 - 3.14 (m, 2 H) 2.77 - 2.93 (m, 2 H) 2.69 (t, J=7.65 Hz, 2 H) 2.12 (br. s., 1 H) 1.95 (br. s., 1 H) 1.43 - 1.72 (m, 3 H). MS (LC/MS) R.T. = 1.10; [M+H]⁺ = 350.10。 Step B: (R) -N- (5-chlorothiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 5-chlorothiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate (100 mg, 0.35 mmol), (S) -3- (aminomethyl) quinuclidine-3 in DMF (1.7 ml) A mixture of -ol dihydrochloride (79 mg, 0.35 mmol) and cesium carbonate (225 mg, 0.69 mmol) was heated at 100 ° C. for 2 h. The reaction mixture was cooled to ambient temperature, poured into water and the solid was collected by filtration. The crude solid was purified by chromatography on silica gel (2-40% 9: 1 methanol: ammonium hydroxide-chloroform) and (R) -N- (5-chlorothiazolo [5,4-b] pyridin-2-yl. ) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine (62 mg, 51% yield).
^{1 H NMR (400 MHz, DMSO} -d 6) δ ppm 9.13 (br. S., 1 H) 7.93 (d, J = 8.53 Hz, 1 H) 7.45 (d, J = 8.28 Hz, 1 H) 3.92 ( d, J = 10.29 Hz, 1 H) 3.67 (d, J = 10.29 Hz, 1 H) 3.00-3.14 (m, 2 H) 2.77-2.93 (m, 2 H) 2.69 (t, J = 7.65 Hz, 2 H) 2.12 (br. S., 1 H) 1.95 (br. S., 1 H) 1.43-1.72 (m, 3 H). MS (LC / MS) RT = 1.10; [M + H] ⁺ = 350.10 .

Example 269
(R) -N ⁵ , N ⁵ -Dimethyl-N ² -(4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-yl) thiazolo [5,4-d] pyrimidine-2,5-diamine

エトキシカルボニル イソチオシアネート(４.３２ml, ３６.６mmol)および２,４−ジクロロ−ピリミジン−５−イルアミン(３.００ｇ, １８.２９mmol)をそのまま混合し、溶解を助けるために５分間超音波処理した。混合物を環境温度で１０分間撹拌し、この時点で全混合物が固化した。メタノール(１００ml)を加え、混合物を３０分間還流し、環境温度まで冷却し、固体を濾過によって集め、５−クロロチアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチルを得た(３.８ｇ, 収率８０％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.64 (s, 1 H) 9.05 (s, 1 H) 4.31 (q, J=7.19 Hz, 2 H) 1.32 (t, J=7.15 Hz, 3 H)。 Step A: Ethyl 5-chlorothiazolo [5,4-d] pyrimidin-2-ylcarbamate

Ethoxycarbonyl isothiocyanate (4.32 ml, 36.6 mmol) and 2,4-dichloro-pyrimidin-5-ylamine (3.00 g, 18.29 mmol) were mixed as is and sonicated for 5 minutes to aid dissolution. . The mixture was stirred at ambient temperature for 10 minutes, at which point the entire mixture solidified. Methanol (100 ml) was added, the mixture was refluxed for 30 minutes, cooled to ambient temperature, and the solid was collected by filtration to give ethyl 5-chlorothiazolo [5,4-d] pyrimidin-2-ylcarbamate (3. 8 g, 80% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 12.64 (s, 1 H) 9.05 (s, 1 H) 4.31 (q, J = 7.19 Hz, 2 H) 1.32 (t, J = 7.15 Hz, 3 H).

５−クロロチアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチル(３００mg, １.１６mmol)を、メタノール中２.０Ｍ ジメチルアミンの溶液(５.０ml, １０.００mmol)に、圧力容器中で懸濁し、この容器を密封し、７５℃の油浴で加熱した。混合物を環境温度まで冷却し、溶媒を蒸発させ、残渣を、水性重炭酸塩とクロロホルムの層間に分配し、３回抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、５−(ジメチルアミノ)チアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチルを得た(２３６mg, 収率９９％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.97 (s, 1 H) 8.67 (s, 1 H) 4.26 (q, J=7.03 Hz, 2 H) 3.17 (s, 6 H) 1.16 - 1.40 (m, 3 H). MS (LC/MS) R.T. = 1.88; [M+H]⁺ = 268.09。 Step B: Ethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbamate

Ethyl 5-chlorothiazolo [5,4-d] pyrimidin-2-ylcarbamate (300 mg, 1.16 mmol) was added to a solution of 2.0 M dimethylamine in methanol (5.0 ml, 10.00 mmol) in a pressure vessel. The vessel was sealed and heated in a 75 ° C. oil bath. The mixture was cooled to ambient temperature, the solvent was evaporated, the residue was partitioned between aqueous bicarbonate and chloroform and extracted three times. The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo to give ethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbamate (236 mg). , Yield 99%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 11.97 (s, 1 H) 8.67 (s, 1 H) 4.26 (q, J = 7.03 Hz, 2 H) 3.17 (s, 6 H) 1.16-1.40 (m, 3 H). MS (LC / MS) RT = 1.88; [M + H] ⁺ = 268.09.

５−(ジメチルアミノ)チアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチル(２３６mg, ０.８８mmol)を、メタノール中２５％(w/w) ナトリウム メトキシドの溶液(５ml, ２３.０mmol)に懸濁し、混合物を一夜還流した。反応混合物を乾固するまで蒸発させ、残渣を水とクロロホルムの層間に分配し、３回抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、エチル Ｎ^５,Ｎ^５−ジメチルチアゾロ[５,４−ｄ]ピリミジン−２,５−ジアミンを白色の固体として得た(１７０mg, 収率９９％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.27 (s, 1 H) 7.44 (s, 2 H) 3.10 (s, 6 H)。 Step C: Ethyl ^{N 5,} N ⁵ - dimethyl-thiazolo [5,4-d] pyrimidine-2,5-diamine

Ethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbamate (236 mg, 0.88 mmol) was added to a solution of 25% (w / w) sodium methoxide in methanol (5 ml, 23.0 mmol). ) And the mixture was refluxed overnight. The reaction mixture was evaporated to dryness and the residue was partitioned between water and chloroform and extracted three times. The combined organics were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo, ethyl N ^5, N ⁵ - dimethyl-thiazolo [5,4-d] pyrimidine-2,5-diamine as a white Obtained as a solid (170 mg, 99% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.27 (s, 1 H) 7.44 (s, 2 H) 3.10 (s, 6 H).

ＤＭＦ(１ml)中のエチル Ｎ^５,Ｎ^５−ジメチルチアゾロ[５,４−ｄ]ピリミジン−２,５−ジアミン(１６０mg, ０.８１９mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(１００μｌ, ２.００mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で二硫化炭素(１２０μｌ, ２mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(１００μｌ, ２.０mmol)を加え、混合物を１０分間再度撹拌した。最後に、ヨードメタン(１２０μｌ, １.９mmol)を滴下した。混合物を５分間撹拌し、この時点で多量の黄色の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、５−(ジメチルアミノ)チアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチルを得た(１９４mg, 収率７９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.75 (s, 1 H) 3.26 (s, 6 H) 2.64 (s, 6 H)。 Step D: Dimethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate

Ethyl N ⁵ in DMF (1ml), ^{N 5} - dimethyl-thiazolo [5,4-d] pyrimidine-2,5-diamine (160mg, 0.819mmol) to a suspension of, 20.0 m sodium hydroxide ( 100 μl, 2.00 mmol) was added. The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (120 μl, 2 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (100 μl, 2.0 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (120 μl, 1.9 mmol) was added dropwise. The mixture was stirred for 5 minutes, at which point a large amount of yellow precipitate had formed. The mixture was poured into water and the solid was collected by filtration to give dimethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (194 mg, 79% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.75 (s, 1 H) 3.26 (s, 6 H) 2.64 (s, 6 H).

ＤＭＦ(１.０ml)中の、５−(ジメチルアミノ)チアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチル(９０mg, ０.３０１mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(８３mg, ０.３６１mmol)および炭酸セシウム(１９６mg, ０.６０mmol)の混合物を、１００℃で１.５時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、粗製の残渣をシリカゲルのクロマトグラフィーによって精製し(２〜４０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、(Ｒ)−Ｎ^５,Ｎ^５−ジメチル−Ｎ^２−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イル)チアゾロ[５,４−ｄ]ピリミジン−２,５−ジアミンを黄褐色の固体として得た(８１mg, 収率７１％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.07 (br. s., 1 H) 8.50 (s, 1 H) 4.01 (d, J=9.54 Hz, 1 H) 3.67 (d, J=9.54 Hz, 1 H) 3.39 (dd, J=14.93, 1.63 Hz, 1 H) 3.23 (s, 6 H) 2.71 - 3.10 (m, 5 H) 2.10 - 2.24 (m, 2 H) 1.68 - 1.84 (m, 1 H) 1.46 - 1.68 (m, 2 H). MS (LC/MS) R.T. = 0.87; [M+H]⁺ = 360.23。 Step ^{E: (R) -N 5,} N 5 - dimethyl -N ² - (4H-1'- azaspiro [oxazole -5,3'- bicyclo [2.2.2] octan] -2-yl) thiazolo [ 5,4-d] pyrimidine-2,5-diamine

Dimethyl 5- (dimethylamino) thiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (90 mg, 0.301 mmol), (S) -3- (aminomethyl) in DMF (1.0 ml). A mixture of quinuclidin-3-ol dihydrochloride (83 mg, 0.361 mmol) and cesium carbonate (196 mg, 0.60 mmol) was heated at 100 ° C. for 1.5 hours. The reaction mixture was cooled to ambient temperature, poured into water and extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered, concentrated in vacuo, and the crude residue was purified by chromatography on silica gel (2-40% 9: 1 methanol: ammonium hydroxide-chloroform). , (R) -N ^{5, N 5} - dimethyl -N ² - (4H-1'- azaspiro [oxazole -5,3'- bicyclo [2.2.2] octan] -2-yl) thiazolo [5, 4-d] pyrimidine-2,5-diamine was obtained as a tan solid (81 mg, 71% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.07 (br.s., 1 H) 8.50 (s, 1 H) 4.01 (d, J = 9.54 Hz, 1 H) 3.67 (d, J = 9.54 Hz, 1 H) 3.39 (dd, J = 14.93, 1.63 Hz, 1 H) 3.23 (s, 6 H) 2.71-3.10 (m, 5 H) 2.10-2.24 (m, 2 H) 1.68-1.84 (m, 1 H ) 1.46-1.68 (m, 2 H). MS (LC / MS) RT = 0.87; [M + H] ⁺ = 360.23.

Example 270
(R) -N-([1,2,4] triazolo [1,5-a] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

ジオキサン(３００ml)中のピラジン−２−アミン(２５ｇ, ２６０mmol)の溶液に、室温で、エトキシカルボニル−イソチオシアネート(３７.９ｇ, ２８９mmol)をゆっくりと加えた。混合物を１８時間撹拌し、溶媒を真空下で蒸発させた。残渣固体をメタノール(１５０ml)およびエタノール(１５０ml)の混合物に溶解した。この溶液に、ＴＥＡ(１０９ml, ７８０mmol)およびヒドロキシルアミン塩酸塩(７２.５ｇ, １０４０mmol)を加えた。混合物を室温で２時間撹拌し、４時間還流した。粗製の混合物を室温まで冷却し、溶媒を蒸発させた。残渣固体をカラムクロマトグラフィーによって精製し(０〜２０％ メタノール／ＣＨ_２Ｃｌ_２)、白色の固体を得た(６０ｇ)。固体をＥｔＯＡｃおよび水に溶かした。水層をＥｔＯＡｃで２回抽出した。合わせた有機層を塩水で洗浄し、硫酸ナトリウムで乾燥させ、[１,２,４]トリアゾロ[１,５−ａ]ピラジン−２−アミンを白色の固体として得た(１２ｇ, ８８mmol, ３３％)。
MS (LCMS) [M+H] = 135.96; R.T. = 0.21分。 Step A: [1,2,4] Triazolo [1,5-a] pyrazin-2-amine

To a solution of pyrazin-2-amine (25 g, 260 mmol) in dioxane (300 ml) was slowly added ethoxycarbonyl-isothiocyanate (37.9 g, 289 mmol) at room temperature. 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 refluxed 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 ₂ Cl ₂ ) to give a white solid (60 g). The solid was dissolved in EtOAc and water. The aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with brine and dried over sodium sulfate to give [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; RT = 0.21 min.

ＤＭＦ(１０ml)中の[１,２,４]トリアゾロ[１,５−ａ]ピラジン−２−アミン(６７６mg, ５mmol)の溶液に、ＮａＯＨ(２０Ｍ, ０.５ml)、ＣＳ_２(１ml)、ＮａＯＨ(２０Ｍ, ０.５ml)およびヨードメタン(１ml)を１０分かけてゆっくりと加えた。混合物を室温で１時間撹拌し、１０mlの水を反応混合物に加えると濁った。混合物をＥｔＯＡｃ(１００ml×３)で抽出した。合わせた有機層を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、蒸発させた。残渣をBiotageシリカゲルカラムで精製し(酢酸エチル−ヘキサン １０〜３０％)、[１,２,４]トリアゾロ[１,５−ａ]ピラジン−２−イルカルボンイミドジチオ酸ジメチルを黄色の固体としてを得た(７２０mg, ３mmol, ６０％)。
¹H NMR (500 MHz, CDCl3) δ 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分。 Step B: [1,2,4] Triazolo [1,5-a] pyrazin-2-ylcarbonimidodithioate dimethyl

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 (20M, 0.5 ml), CS ₂ (1 ml), NaOH (20M, 0.5 ml) and iodomethane (1 ml) were added slowly over 10 minutes. The mixture was stirred at room temperature for 1 hour and became cloudy when 10 ml of water was added to the reaction mixture. The mixture was extracted with EtOAc (100 ml x 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%) and dimethyl [1,2,4] triazolo [1,5-a] pyrazin-2-ylcarbonimidodithioate as a yellow solid. Obtained (720 mg, 3 mmol, 60%).
¹ H NMR (500 MHz, CDCl3) δ 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; RT = 1.55 minutes.

ＤＭＦ(５ml)中の、[１,２,４]トリアゾロ[１,５−ａ]ピラジン−２−イルカルボンイミドジチオ酸ジメチル(１２０mg, ０.５０mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１２０mg, ０.７６mmol)および炭酸セシウム(４９２mg, １.５mmol)の混合物を、７０℃で６時間加熱した。混合物を濃縮し、Biotageシリカゲルカラムで精製し(１００％ 酢酸エチル、次に１０〜３５％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、望ましい生成物である(Ｒ)−Ｎ−([１,２,４]トリアゾロ[１,５−ａ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の固体として得た(８５mg, ２６.７％)。
¹H NMR (500 MHz, MeOD) δ 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). MS (LCMS) [M+H] = 300.06; R.T. = 0.2分。 Step C: (R) -N-([1,2,4] triazolo [1,5-a] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2. 2.2] Octane] -2-amine

Dimethyl [1,2,4] triazolo [1,5-a] pyrazin-2-ylcarbonimidodithioate (120 mg, 0.50 mmol), (S) -3- (aminomethyl) in DMF (5 ml) A mixture of quinuclidin-3-ol dihydrochloride (120 mg, 0.76 mmol) and cesium carbonate (492 mg, 1.5 mmol) was heated at 70 ° 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), the desired product (R) -N-([1, 2,4] triazolo [1,5-a] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Obtained as a solid (85 mg, 26.7%).
¹ H NMR (500 MHz, MeOD) δ 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). MS (LCMS) [M + H] = 300.06; RT = 0.2 min.

Example 271
(R) -N- (thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ＤＭＦ(２ml)中のチアゾロ[５,４−ｂ]ピリジン−２−アミン(３００mg, １.９８mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(２００μｌ, ４.０mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で二硫化炭素(３００μｌ, ４.９６mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(２００μｌ, ４.０mmol)を加え、混合物を再度１０分間撹拌した。最後に、ヨードメタン(３００μｌ, ４.７６mmol)を滴下した。混合物を５分間撹拌し、この時点で、多量の黄色の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、さらに精製することなく用いるのに十分な純度の黄色の固体として、チアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルを得た(１９０mg, 収率３８％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 8.47 (d, J=4.58 Hz, 1 H) 8.11 (dd, J=8.24, 1.53 Hz, 1 H) 7.37 (dd, J=8.24, 4.88 Hz, 1 H) 2.66 (s, 6 H)。 Step A: Dimethyl thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate

To a suspension of thiazolo [5,4-b] pyridin-2-amine (300 mg, 1.98 mmol) in DMF (2 ml) was added 20.0 M sodium hydroxide (200 μl, 4.0 mmol). The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (300 μl, 4.96 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (200 μl, 4.0 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (300 μl, 4.76 mmol) was added dropwise. The mixture was stirred for 5 minutes, at which point a large amount of yellow precipitate had formed. The mixture was poured into water and the solid was collected by filtration to give dimethyl thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate as a yellow solid of sufficient purity to be used without further purification. (190 mg, 38% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 8.47 (d, J = 4.58 Hz, 1 H) 8.11 (dd, J = 8.24, 1.53 Hz, 1 H) 7.37 (dd, J = 8.24, 4.88 Hz, 1 H) 2.66 (s, 6 H).

ＤＭＦ(１ml)中の、チアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(９０mg, ０.３５mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(９７mg, ０.４２mmol)および炭酸セシウム(２３０mg, ０.７１mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。混合物をシリカゲルのクロマトグラフィーによって精製した(２〜４０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)。生成物のフラクションを合わせ、真空で濃縮し、(Ｒ)−Ｎ−(チアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(８４mg, 収率７６％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.30 (br. s., 1 H) 8.37 (dd, J=4.77, 1.51 Hz, 1 H) 7.82 (dd, J=8.03, 1.51 Hz, 1 H) 7.28 (dd, J=8.03, 4.77 Hz, 1 H) 4.05 (d, J=9.54 Hz, 1 H) 3.70 (d, J=9.54 Hz, 1 H) 3.42 (dd, J=15.06, 1.76 Hz, 1 H) 2.75 - 3.07 (m, 5 H) 2.14 - 2.26 (m, 2 H) 1.71 - 1.84 (m, J=13.99, 9.79, 4.17, 4.17 Hz, 1 H) 1.48 - 1.68 (m, 2 H). MS (LC/MS) R.T. = 0.64; [M+H]⁺ = 316.15。 Step B: (R) -N- (thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- 2-Amine

Dimethyl thiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate (90 mg, 0.35 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride in DMF (1 ml) A mixture of salt (97 mg, 0.42 mmol) and cesium carbonate (230 mg, 0.71 mmol) was heated at 100 ° C. for 2 hours. The reaction mixture was cooled to ambient temperature, poured into water and extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The mixture was purified by chromatography on silica gel (2-40% 9: 1 methanol: ammonium hydroxide-chloroform). The product fractions were combined, concentrated in vacuo, and (R) -N- (thiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine was obtained (84 mg, 76% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.30 (br. S., 1 H) 8.37 (dd, J = 4.77, 1.51 Hz, 1 H) 7.82 (dd, J = 8.03, 1.51 Hz, 1 H) 7.28 (dd, J = 8.03, 4.77 Hz, 1 H) 4.05 (d, J = 9.54 Hz, 1 H) 3.70 (d, J = 9.54 Hz, 1 H) 3.42 (dd, J = 15.06, 1.76 Hz, 1 H) 2.75-3.07 (m, 5 H) 2.14-2.26 (m, 2 H) 1.71-1.84 (m, J = 13.99, 9.79, 4.17, 4.17 Hz, 1 H) 1.48-1.68 (m, 2 H). MS (LC / MS) RT = 0.64; [M + H] ⁺ = 316.15.

(Ｒ)−Ｎ−(６−ブロモチアゾロ[５,４−ｂ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(４４mg, ０.１１１mmol)を、ＭｅＯＨ(５０ml)に懸濁し、全ての固体が溶解するまで３Ｎ ＨＣｌを加えた(約１０ml)。反応フラスコに窒素を吹きつけ、１０％ パラジウム／炭素(３５mg)を加え、フラスコに水素バルーンを取り付けた。混合物を一夜反応させ、この時点でＴＬＣにより出発物質が消費されたことが示された。フラスコに窒素を吹きつけ、セライトで濾過し、メタノールで洗浄した。合わせた濾液を約９０％まで濃縮して、大部分のメタノールを除去し、重炭酸ナトリウムの飽和溶液を添加することによって、溶液を塩基性にした。塩基性の水相をクロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。混合物をシリカゲルのクロマトグラフィーによって精製した(２〜４０％ [９：１ メタノール：水酸化アンモニウム]−クロロホルム)。生成物のフラクションを合わせ、真空で濃縮し、(Ｒ)−Ｎ−(チアゾロ[５,４−ｂ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２４mg, ０.０７５mmol, 収率６７.５％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.50 (br. s., 1 H) 8.41 (d, J=2.76 Hz, 1 H) 8.27 (d, J=2.76 Hz, 1 H) 4.06 (d, J=9.79 Hz, 1 H) 3.72 (d, J=9.79 Hz, 1 H) 3.43 (dd, J=15.06, 1.76 Hz, 1 H) 2.74 - 3.09 (m, 5 H) 2.12 - 2.25 (m, 2 H) 1.71 - 1.86 (m, 1 H) 1.49 - 1.67 (m, 2 H). MS (LC/MS) R.T. = 0.75; [M+H]⁺ = 317.13。 Example 272
(R) -N- (thiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-bromothiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 Amine (44 mg, 0.111 mmol) was suspended in MeOH (50 ml) and 3N HCl was added until all solids were dissolved (ca. 10 ml). Nitrogen was bubbled through the reaction flask, 10% palladium / carbon (35 mg) was added, and a hydrogen balloon was attached to the flask. The mixture was allowed to react overnight, at which point TLC showed the starting material was consumed. The flask was flushed with nitrogen, filtered through celite, and washed with methanol. The combined filtrate was concentrated to about 90% to remove most of the methanol and the solution was made basic by adding a saturated solution of sodium bicarbonate. The basic aqueous phase was extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The mixture was purified by chromatography on silica gel (2-40% [9: 1 methanol: ammonium hydroxide] -chloroform). The product fractions were combined, concentrated in vacuo, and (R) -N- (thiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine was obtained (24 mg, 0.075 mmol, 67.5% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.50 (br.s., 1 H) 8.41 (d, J = 2.76 Hz, 1 H) 8.27 (d, J = 2.76 Hz, 1 H) 4.06 (d, J = 9.79 Hz, 1 H) 3.72 (d, J = 9.79 Hz, 1 H) 3.43 (dd, J = 15.06, 1.76 Hz, 1 H) 2.74-3.09 (m, 5 H) 2.12-2.25 (m, 2 H) 1.71-1.86 (m, 1 H) 1.49-1.67 (m, 2 H). MS (LC / MS) RT = 0.75; [M + H] ⁺ = 317.13.

Example 273
(R) -N- (7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] Octane] -2-amine

４,６−ジクロロ−２−メチルピリミジン−５−アミン(１ｇ, ５.６２mmol)およびＯ−エチル カルボンイソチオシアナチデート(O-ethyl carbonisothiocyanatidate)(０.６６ml, ５.６２mmol)の混合物に、トルエン(２ml)を加え、固体を完全に湿らせた。混合物を１００℃の油浴上に１.５時間置き、この時点で混合物が固着し(seized)、固体を得た。固体を環境温度まで冷却し、エーテルで磨砕し、得られた固体を濾過によって集め、７−クロロ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチルを得た(１.０８ｇ, ３.９６mmol, 収率７０.５％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.70 (br. s., 1 H) 4.30 (q, J=7.19 Hz, 2 H) 2.69 (s, 3 H) 1.30 (t, J=7.15 Hz, 3 H)。 Step A: Ethyl 7-chloro-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbamate

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) was added to completely wet the solid. The mixture was placed on a 100 ° C. oil bath for 1.5 hours, at which point the mixture seized and a solid was obtained. The solid was cooled to ambient temperature, triturated with ether, and the resulting solid was collected by filtration to give ethyl 7-chloro-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbamate (1 0.08 g, 3.96 mmol, yield 70.5%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 12.70 (br. S., 1 H) 4.30 (q, J = 7.19 Hz, 2 H) 2.69 (s, 3 H) 1.30 (t, J = 7.15 Hz, 3 H).

７−クロロ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イルカルバミン酸エチル(３００mg, １.１００mmol)を、メタノール中２５％(w/w)のナトリウム メトキシドの溶液(５ml, ２３.１４mmol)に懸濁し、混合物を一夜還流した。混合物を環境温度まで冷却し、水で希釈し、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮し、７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−アミンを得た(１２０mg, ０.６１２mmol, 収率５５.６％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.71 (s, 2 H) 3.98 (s, 3 H) 2.52 (s, 3 H)。 Step B: 7-Methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-amine

Ethyl 7-chloro-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbamate (300 mg, 1.100 mmol) was added to a solution of 25% (w / w) 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 (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo to give 7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-amine (120 mg, 0. 612 mmol, yield 55.6%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.71 (s, 2 H) 3.98 (s, 3 H) 2.52 (s, 3 H).

ＤＭＦ(０.５ml)中の７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−アミン(１００mg, ０.５１mmol)の懸濁液に、１６.０Ｍ 水酸化ナトリウム(７５μｌ, １.２mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で、二硫化炭素(８０μｌ, １.２７mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに１６.０Ｍ 水酸化ナトリウム(７５μｌ, １.２mmol)を加え、混合物を再度１０分間撹拌した。最後にヨードメタン(８０μｌ, １.２９mmol)を滴下した。混合物を５分間撹拌し、この時点で、多量の黄色の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、粗製の黄色の固体を得た。これをさらにシリカゲルのクロマトグラフィーによって精製した(２〜２０％ 酢酸エチル−クロロホルム)。生成物のフラクションを合わせて、真空で濃縮し、７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチルを黄色の固体として得た。(９０mg, 収率５９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 4.17 (s, 3 H) 2.71 (s, 3 H) 2.64 (s, 6 H)。 Step C: 7-Methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate dimethyl ester

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 16.0 M sodium hydroxide (75 μl, 1 0.2 mmol) was added. The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (80 μl, 1.27 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. A further 16.0M sodium hydroxide (75 μl, 1.2 mmol) was added and the mixture was stirred again for 10 minutes. Finally iodomethane (80 μl, 1.29 mmol) was added dropwise. The mixture was stirred for 5 minutes, at which point a large amount of yellow precipitate had formed. The mixture was poured into water and the solid was collected by filtration to give a crude yellow solid. This was further purified by chromatography on silica gel (2-20% ethyl acetate-chloroform). The product fractions were combined and concentrated in vacuo to give dimethyl 7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate as a yellow solid. (90 mg, 59% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 4.17 (s, 3 H) 2.71 (s, 3 H) 2.64 (s, 6 H).

ＤＭＦ(０.５ml)中の７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチル(５６mg, ０.１９mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(５１mg, ０.２２mmol)および炭酸セシウム(１７５mg, ０.５４mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。混合物をシリカゲルのクロマトグラフィーによって精製した(２〜４０％ [９：１ メタノール：水酸化アンモニウム]−クロロホルム)。生成物のフラクションを合わせ、真空で濃縮し、(Ｒ)−Ｎ−(７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(３４mg, 収率５０％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.10 (br. s., 1 H) 4.14 (s, 3 H) 4.03 (d, J=9.54 Hz, 1 H) 3.68 (d, J=9.54 Hz, 1 H) 3.39 (dd, J=14.93, 1.63 Hz, 1 H) 2.74 - 3.07 (m, 5 H) 2.68 (s, 3 H) 2.04 - 2.28 (m, 2 H) 1.70 - 1.86 (m, 1 H) 1.44 - 1.67 (m, 2 H). MS (LC/MS) R.T. = 1.10; [M+H]⁺ = 361.32。 Step D: (R) -N- (7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2 .2] Octane] -2-amine

Dimethyl 7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (56 mg, 0.19 mmol), (S) -3- (aminomethyl) in DMF (0.5 ml) A mixture of quinuclidin-3-ol dihydrochloride (51 mg, 0.22 mmol) and cesium carbonate (175 mg, 0.54 mmol) was heated at 100 ° C. for 2 hours. The reaction mixture was cooled to ambient temperature, poured into water and extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The mixture was purified by chromatography on silica gel (2-40% [9: 1 methanol: ammonium hydroxide] -chloroform). The product fractions were combined, concentrated in vacuo, and (R) -N- (7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5 , 3′-bicyclo [2.2.2] octane] -2-amine was obtained (34 mg, 50% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.10 (br. S., 1 H) 4.14 (s, 3 H) 4.03 (d, J = 9.54 Hz, 1 H) 3.68 (d, J = 9.54 Hz, 1 H) 3.39 (dd, J = 14.93, 1.63 Hz, 1 H) 2.74-3.07 (m, 5 H) 2.68 (s, 3 H) 2.04-2.28 (m, 2 H) 1.70-1.86 (m, 1 H ) 1.44-1.67 (m, 2 H). MS (LC / MS) RT = 1.10; [M + H] ⁺ = 361.32.

Example 274
(R) -N- (7-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ＤＭＦ(１.５ml)中の７−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−アミン(３００mg, １.６７mmol)の懸濁液に、１６.０Ｍ 水酸化ナトリウム(２１０μｌ, ３.４mmol)を加えた。混合物を室温で１０分間撹拌し、この時点で二硫化炭素(２５０μｌ, ４.１５mmol)を加え、得られた帯赤褐色の混合物を１０分間撹拌した。さらに１６.０Ｍ 水酸化ナトリウム(２１０μｌ, ３.４mmol)を加え、混合物を再度１０分間撹拌した。最後にヨードメタン(２５０μｌ, ４.００mmol)を滴下した。混合物を１０分間撹拌し、この時点で多量の黄色の沈殿物が形成した。物質を水に注ぎ、固体を濾過によって集め、７−メトキシ−５−メチルチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチ酸ジメチルを黄色の固体として得た(３２４mg, 収率６９％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 8.60 (s, 1 H) 4.20 (s, 3 H) 2.65 (s, 6 H)。 Step A: Dimethyl 7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate

To a suspension of 7-methoxythiazolo [5,4-d] pyrimidin-2-amine (300 mg, 1.67 mmol) in DMF (1.5 ml) was added 16.0 M sodium hydroxide (210 μl, 3.4 mmol). ) Was added. The mixture was stirred at room temperature for 10 minutes, at which point carbon disulfide (250 μl, 4.15 mmol) was added and the resulting reddish brown mixture was stirred for 10 minutes. A further 16.0M sodium hydroxide (210 μl, 3.4 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (250 μl, 4.00 mmol) was added dropwise. The mixture was stirred for 10 minutes, at which point a large amount of yellow precipitate had formed. The material was poured into water and the solid was collected by filtration to give dimethyl 7-methoxy-5-methylthiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate as a yellow solid (324 mg, 69% yield). ).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 8.60 (s, 1 H) 4.20 (s, 3 H) 2.65 (s, 6 H).

ＤＭＦ(３ml)中の、７−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−イルカルボンイミドジチオ酸ジメチル(１５０mg, ０.５２mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１３２mg, ０.５８mmol)および炭酸セシウム(４２７mg, １.３１mmol)の混合物を、１００℃で２時間加熱した。反応混合物を環境温度まで冷却し、水に注ぎ、クロロホルム(４×)で抽出した。合わせた有機物を塩水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。混合物をシリカゲルのクロマトグラフィーによって精製した(２〜４０％ [９：１ メタノール：水酸化アンモニウム]−クロロホルム)。生成物のフラクションを合わせ、真空で濃縮し、(Ｒ)−Ｎ−(７−メトキシチアゾロ[５,４−ｄ]ピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(９５mg, 収率５１％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.12 (br. s., 1 H) 8.52 (s, 1 H) 4.16 (s, 3 H) 4.05 (d, J=9.54 Hz, 1 H) 3.70 (d, J=9.54 Hz, 1 H) 3.40 (dd, J=14.93, 1.88 Hz, 1 H) 2.70 - 3.07 (m, 5 H) 2.08 - 2.27 (m, 2 H) 1.68 - 1.85 (m, 1 H) 1.48 - 1.66 (m, 2 H). MS (LC/MS) R.T. = 0.90; [M+H]⁺ = 347.34。 Step B: (R) -N- (7-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

Dimethyl 7-methoxythiazolo [5,4-d] pyrimidin-2-ylcarbonimidodithioate (150 mg, 0.52 mmol), (S) -3- (aminomethyl) quinuclidine-3 in DMF (3 ml) A mixture of -ol dihydrochloride (132 mg, 0.58 mmol) and cesium carbonate (427 mg, 1.31 mmol) was heated at 100 ° C. for 2 hours. The reaction mixture was cooled to ambient temperature, poured into water and extracted with chloroform (4x). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The mixture was purified by chromatography on silica gel (2-40% [9: 1 methanol: ammonium hydroxide] -chloroform). The product fractions were combined, concentrated in vacuo and (R) -N- (7-methoxythiazolo [5,4-d] pyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3 '-Bicyclo [2.2.2] octane] -2-amine was obtained (95 mg, 51% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.12 (br. S., 1 H) 8.52 (s, 1 H) 4.16 (s, 3 H) 4.05 (d, J = 9.54 Hz, 1 H) 3.70 ( d, J = 9.54 Hz, 1 H) 3.40 (dd, J = 14.93, 1.88 Hz, 1 H) 2.70-3.07 (m, 5 H) 2.08-2.27 (m, 2 H) 1.68-1.85 (m, 1 H ) 1.48-1.66 (m, 2 H). MS (LC / MS) RT = 0.90; [M + H] ⁺ = 347.34.

(Ｒ)−２−(４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−イルアミノ)チアゾール−５−カルボニトリルを、実施例２７４の方法によって、２−アミノ−５−シアノチアゾールから出発して合成した。
¹H NMR (500 MHz, DMSO-D6) δ ppm 9.05 (s, 1 H), 8.13 (s, 1 H), 3.86 (d, J=10.38 Hz, 1 H), 3.61 (d, J=10.38 Hz, 1 H), 3.01 - 3.10 (m, 2 H), 2.83 (t, J=7.63 Hz, 2 H), 2.62 - 2.71 (m, 2 H), 2.09 (s, 1 H), 1.90 - 1.97 (m, 2 H), 1.54 - 1.62 (m, 3 H). MS (LC/MS) R.T. = 0.52; [M+H]⁺ = 290.0。 Example 275
(R) -2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) thiazole-5-carbonitrile

(R) -2- (4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-ylamino) thiazole-5-carbonitrile was prepared by the method of Example 274. Synthesized starting from 2-amino-5-cyanothiazole.
¹ H NMR (500 MHz, DMSO-D6) δ ppm 9.05 (s, 1 H), 8.13 (s, 1 H), 3.86 (d, J = 10.38 Hz, 1 H), 3.61 (d, J = 10.38 Hz , 1 H), 3.01-3.10 (m, 2 H), 2.83 (t, J = 7.63 Hz, 2 H), 2.62-2.71 (m, 2 H), 2.09 (s, 1 H), 1.90-1.97 ( m, 2 H), 1.54-1.62 (m, 3 H). MS (LC / MS) RT = 0.52; [M + H] ⁺ = 290.0.

(Ｒ)−Ｎ−(７−ブロモピロロ[１,２−ｆ][１,２,４]トリアジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７４の方法によって、７−ブロモピロロ[１,２−ｆ][１,２,４]トリアジン−４−アミンから出発して合成した。
¹H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, s), 7.04 (1 H, d, J=4.53 Hz), 6.77 (1 H, d, J=4.53 Hz), 4.09 (1 H, d, J=10.32 Hz), 3.79 (1 H, d, J=10.58 Hz), 3.24 (1 H, d), 3.12 (1 H, d), 2.70 - 3.00 (4 H, m), 2.06 - 2.25 (2 H, m), 1.52 - 1.86 (3 H, m). MS (LC/MS) R.T. = 1.62; [M+H]⁺ = 377.2。 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] Octane] -2-amine

(R) -N- (7-bromopyrrolo [1,2-f] [1,2,4] triazin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2 .2] Octane] -2-amine was synthesized by the method of Example 274 starting from 7-bromopyrrolo [1,2-f] [1,2,4] triazine-4-amine.
¹ H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, s), 7.04 (1 H, d, J = 4.53 Hz), 6.77 (1 H, d, J = 4.53 Hz), 4.09 (1 H, d, J = 10.32 Hz), 3.79 (1 H, d, J = 10.58 Hz), 3.24 (1 H, d), 3.12 (1 H, d), 2.70-3.00 (4 H, m), 2.06-2.25 (2 H, m), 1.52-1.86 (3 H, m). MS (LC / MS) RT = 1.62; [M + H] ⁺ = 377.2.

(Ｒ)−Ｎ−(１,６−ナフチリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７４の方法によって、１,６−ナフチリジン−２−アミンから出発して合成した。
¹H NMR (400 MHz, MeOD) δ ppm 8.99 (1 H, s), 8.48 (1 H, d, J=6.04 Hz), 8.20 (1 H, d, J=8.56 Hz), 7.77 (1 H, d, J=6.04 Hz), 7.12 (1 H, d, J=8.81 Hz), 4.12 (1 H, d, J=10.32 Hz), 3.82 (1 H, d, J=10.32 Hz), 3.36 (1 H, d), 3.21 (1 H, d), 2.79 - 3.09 (4 H, m), 2.08 - 2.30 (2 H, m), 1.56 - 1.95 (3 H, m). (LC/MS) R.T. = 0.38; [M+H]⁺ = 310.3。 Example 277
(R) -N- (1,6-Naphthyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (1,6-naphthyridin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared as an example. Synthesized by the method of 274 starting from 1,6-naphthyridin-2-amine.
¹ H NMR (400 MHz, MeOD) δ ppm 8.99 (1 H, s), 8.48 (1 H, d, J = 6.04 Hz), 8.20 (1 H, d, J = 8.56 Hz), 7.77 (1 H, d, J = 6.04 Hz), 7.12 (1 H, d, J = 8.81 Hz), 4.12 (1 H, d, J = 10.32 Hz), 3.82 (1 H, d, J = 10.32 Hz), 3.36 (1 H, d), 3.21 (1 H, d), 2.79-3.09 (4 H, m), 2.08-2.30 (2 H, m), 1.56-1.95 (3 H, m). (LC / MS) RT = 0.38; [M + H] ⁺ = 310.3.

(Ｒ)−Ｎ−(キナゾリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７４の方法によって、２−アミノキナゾリンから出発して合成した。
¹H NMR (400 MHz, MeOD) δ ppm 9.27 (1 H, s), 7.72 - 7.99 (3 H, m), 7.47 (1 H, dd, J=7.55, 3.78 Hz), 4.07 (1 H, d, J=10.07 Hz), 3.76 (1 H, d, J=10.07 Hz), 3.26 (1 H, br. s.), 3.13 (1 H, d), 2.70 - 3.03 (4 H, m), 2.17 (2 H, br. s.), 1.50 - 1.88 (3 H, m). (LC/MS) R.T. = 1.11; [M+H]⁺ = 310.3。 Example 278
(R) -N- (Quinazolin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (Quinazolin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared by the method of Example 274. Synthesized starting from 2-aminoquinazoline.
¹ H NMR (400 MHz, MeOD) δ ppm 9.27 (1 H, s), 7.72-7.99 (3 H, m), 7.47 (1 H, dd, J = 7.55, 3.78 Hz), 4.07 (1 H, d , J = 10.07 Hz), 3.76 (1 H, d, J = 10.07 Hz), 3.26 (1 H, br. S.), 3.13 (1 H, d), 2.70-3.03 (4 H, m), 2.17 (2 H, br. S.), 1.50-1.88 (3 H, m). (LC / MS) RT = 1.11; [M + H] ⁺ = 310.3.

Example 279
(R) -N- (6,8-Dichloroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(２,４−ジクロロフェニル)メタンアミン(２ｇ, １１.４mmol)を、メタノール(１０ml)中の２,２−ジエトキシアセトイミド酸メチル(２.０４ｇ, １２.６mmol)の溶液に加えた。混合物を７０℃で１時間加熱した。混合物をクロマトグラフィーによって精製した(Biotage, １００％ 酢酸エチル)。望ましいフラクションを濃縮し、Ｎ−(２,４−ジクロロベンジル)−２,２−ジエトキシアセトイミドアミドを無色の粘性の油状物として得た(２.８ｇ, ９.２mmol, 収率７２.７％)。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 7.27 - 7.70 (m, 3 H), 4.77 (s, 1 H), 4.14 - 4.35 (m, 2 H), 3.45 - 3.68 (m, 4 H), 1.09 - 1.29 (m, 6 H). LC/MS RT=2.03; [M+H]⁺ = 304.9。 Step A: N- (2,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide

(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 ° C. for 1 hour. The mixture was purified by chromatography (Biotage, 100% ethyl acetate). The desired fraction was concentrated to give N- (2,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide as a colorless viscous oil (2.8 g, 9.2 mmol, 72.7 yield). %).
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 7.27-7.70 (m, 3 H), 4.77 (s, 1 H), 4.14-4.35 (m, 2 H), 3.45-3.68 (m, 4 H ), 1.09-1.29 (m, 6 H). LC / MS RT = 2.03; [M + H] ⁺ = 304.9.

硫酸(４ml, ７５mmol)に、Ｎ−(２,４−ジクロロベンジル)−２,２−ジエトキシアセトイミドアミド(２ｇ, ６.６mmol)を室温で加えた。反応物を４０℃で１８時間加熱した。ＴＬＣおよびＬＣ／ＭＳにより、生成物の存在が示された。反応物を室温まで冷却し、反応混合物がｐＨ 約７になるまで、水性ＮａＯＨ(約１５Ｍ)で反応停止させた。粗生成物を酢酸エチル(２×５０ml)で抽出し、有機物をＭｇＳＯ_４で乾燥させ、濾過し、真空で濃縮し、生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, １０〜８０％ 酢酸エチル／ヘキサン)、５,７−ジクロロイソキノリン−１−アミンを暗黄色の粉末として得た(０.３２ｇ, １.５０mmol, 収率２２.９％)。
¹H NMR (400 MHz, DMSO-D₆) δ ppm 8.99 (s, 1 H), 7.64 - 7.73 (m, 1 H), 7.30 (d, J=2.01 Hz, 1 H), 6.61 (s, 1 H), 6.43 (s, 2 H). MS (LC/MS) R.T. = 1.40; [M+H]⁺ = 213.1。 Step B: 6,8-Dichloroisoquinolin-3-amine

To sulfuric acid (4 ml, 75 mmol), N- (2,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide (2 g, 6.6 mmol) was added at room temperature. The reaction was heated at 40 ° C. for 18 hours. TLC and LC / MS indicated the presence of product. The reaction was cooled to room temperature and quenched with aqueous NaOH (about 15M) until the reaction mixture was about pH 7. The crude product was extracted with ethyl acetate (2 × 50 ml) and the organics were dried over MgSO ₄ , filtered and concentrated in vacuo to give the product. The crude product was purified by chromatography (Biotage, 10-80% ethyl acetate / hexane) to give 5,7-dichloroisoquinolin-1-amine as a dark yellow powder (0.32 g, 1.50 mmol, yield). Rate 22.9%).
¹ H NMR (400 MHz, DMSO-D ₆ ) δ ppm 8.99 (s, 1 H), 7.64-7.73 (m, 1 H), 7.30 (d, J = 2.01 Hz, 1 H), 6.61 (s, 1 H), 6.43 (s, 2 H). MS (LC / MS) RT = 1.40; [M + H] ⁺ = 213.1.

ジクロロメタン(２０ml)中の６,８−ジクロロイソキノリン−３−アミン(０.２７ｇ, １.２８mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.３０ｇ, １.２９mmol)を加え、反応混合物を４０℃で４時間撹拌した。反応物を室温まで冷却し、クロマトグラフィーを行い(Biotage, １０〜１００％ 酢酸エチル／ヘキサン)、６,８−ジクロロ−３−イソチオシアナトイソキノリンを粉末として得た(０.２ｇ, ０.７８mmol, 収率６１.９％)。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 9.42 (s, 1 H), 8.16 (d, J=1.83 Hz, 1 H), 8.02 (d, J=2.14 Hz, 1 H), 7.92 (s, 1 H). MS (LC/MS) R.T. = 3.63; [M+H]⁺ = 255.0。 Step C: 6,8-Dichloro-3-isothiocyanatoisoquinoline

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, 1.30 mmol). 29 mmol) was added and the reaction mixture was stirred at 40 ° C. for 4 h. The reaction was cooled to room temperature and chromatographed (Biotage, 10-100% ethyl acetate / hexanes) to give 6,8-dichloro-3-isothiocyanatoisoquinoline as a powder (0.2 g, 0.78 mmol). Yield 61.9%).
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 9.42 (s, 1 H), 8.16 (d, J = 1.83 Hz, 1 H), 8.02 (d, J = 2.14 Hz, 1 H), 7.92 ( s, 1 H). MS (LC / MS) RT = 3.63; [M + H] ⁺ = 255.0.

ＤＭＦ(１０ml)中の６,８−ジクロロ−３−イソチオシアナトイソキノリン(０.１７ｇ, ０.６７mmol)に、炭酸セシウム(０.５４３ｇ, １.６７mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１５ｇ, ０.６７mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。生成物をＮ,Ｎ'−ジイソプロピルカルボジイミド(０.３１ml, ２.０mmol)で処理した。反応物を９０℃で４時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物を少量の酢酸エチルに溶かすと、沈殿物が形成した。それを濾過して取り、少量の酢酸エチルで洗浄し、真空オーブン中で乾燥させ、(Ｒ)−Ｎ−(６,８−ジクロロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０９４ｇ, ０.２４mmol, 収率３６.６％)。
¹H NMR (400 MHz, DMSO-D₆) δ ppm 9.23 (s, 1 H), 8.71 - 8.83 (m, 1 H), 7.90 - 8.00 (m, 1 H), 7.57 - 7.67 (m, 1 H), 7.13 - 7.24 (m, 1 H), 3.79 - 3.90 (m, 1 H), 3.53 - 3.64 (m, 1 H), 2.93 - 3.04 (m, 2 H), 2.72 - 2.82 (m, 2 H), 2.61 - 2.70 (m, 2 H), 1.99 (s, 1 H), 1.90 (s, 1 H), 1.59 (d, J=4.78 Hz, 2 H), 1.40 - 1.50 (m, 1 H). MS (LC/MS) R.T. = 1.68; [M+H]⁺ = 377.1。 Step D: (R) -N- (6,8-Dichloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

6,8-Dichloro-3-isothiocyanatoisoquinoline (0.17 g, 0.67 mmol) in DMF (10 ml) was added to cesium carbonate (0.543 g, 1.67 mmol) and (S) -3- (aminomethyl). Quinuclidin-3-ol dihydrochloride (0.15 g, 0.67 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 at 90 ° C. for 4 hours. The reaction was cooled to room temperature and concentrated to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH). The product was dissolved in a small amount of ethyl acetate and a precipitate formed. It is filtered off, washed with a small amount of ethyl acetate, dried in a vacuum oven and (R) -N- (6,8-dichloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole- 5,3′-Bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (0.094 g, 0.24 mmol, 36.6% yield).
¹ H NMR (400 MHz, DMSO-D ₆ ) δ ppm 9.23 (s, 1 H), 8.71-8.83 (m, 1 H), 7.90-8.00 (m, 1 H), 7.57-7.67 (m, 1 H ), 7.13-7.24 (m, 1 H), 3.79-3.90 (m, 1 H), 3.53-3.64 (m, 1 H), 2.93-3.04 (m, 2 H), 2.72-2.82 (m, 2 H ), 2.61-2.70 (m, 2 H), 1.99 (s, 1 H), 1.90 (s, 1 H), 1.59 (d, J = 4.78 Hz, 2 H), 1.40-1.50 (m, 1 H) MS (LC / MS) RT = 1.68; [M + H] ⁺ = 377.1.

Example 280

(R) -N- (6,7-Dichloroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(３,４−ジクロロフェニル)メタンアミン(２ｇ, １１.４mmol)を、メタノール(１０ml)中の２,２−ジエトキシアセトイミド酸メチル(２.０４ｇ, １２.６mmol)の溶液に加えた。混合物を７０℃で１時間加熱した。混合物をクロマトグラフィーによって精製した(Biotage, １００％ 酢酸エチル)。望ましいフラクションを濃縮し、Ｎ−(３,４−ジクロロベンジル)−２,２−ジエトキシアセトイミドアミドを無色の粘性の油状物として得た(２.８ｇ, ９.２mmol, 収率７２.７％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 7.45 (m, 1 H), 7.40 (m, 1 H), 7.19 (dd, J=8.09, 1.98 Hz, 1 H), 4.94 (s, 1 H), 4.43 (s, 2 H), 3.47 - 3.77 (m, 4 H), 1.41 - 1.79 (m, 6 H). LC/MS RT=2.15; [M+H]⁺ = 305.1。 Step A: N- (3,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide

(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 ° C. for 1 hour. The mixture was purified by chromatography (Biotage, 100% ethyl acetate). The desired fraction was concentrated to give N- (3,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide as a colorless viscous oil (2.8 g, 9.2 mmol, 72.7 yield). %).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 7.45 (m, 1 H), 7.40 (m, 1 H), 7.19 (dd, J = 8.09, 1.98 Hz, 1 H), 4.94 (s, 1 H) , 4.43 (s, 2 H), 3.47-3.77 (m, 4 H), 1.41-1.79 (m, 6 H). LC / MS RT = 2.15; [M + H] ⁺ = 305.1.

硫酸(４ml, ７５mmol)に、Ｎ−(３,４−ジクロロベンジル)−２,２−ジエトキシアセトイミドアミド(２ｇ, ６.６mmol)を室温で加えた。反応物を４０℃で４９時間加熱した。ＴＬＣおよびＬＣ／ＭＳにより、生成物の存在が示された。反応物を室温まで冷却し、反応混合物がｐＨ 約７になるまで水性ＮａＯＨ(約１５Ｍ)で反応停止させた。粗生成物を酢酸エチル(２×５０ml)で抽出し、有機物をＭｇＳＯ_４で乾燥させ、濾過し、真空で濃縮し、生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, １００％ 酢酸エチルから[９０／１０％ 酢酸エチル／ＭｅＯＨ])、位置異性体６,７−ジクロロイソキノリン−１−アミンおよび５,６−ジクロロイソキノリン−３−アミンの約１：１混合物を、暗黄色の粉末として得た(１.２ｇ, ５.６４mmol, 収率８６.０％)。位置異性体を分離せずに用いた。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 8.90 (s, 1 H), 8.83 (s, 1 H), 8.12 (s, 1 H), 7.89 (s, 1 H), 7.84 (d, J=8.54 Hz, 1 H), 7.28 (d, J=8.85 Hz, 1 H), 6.83 (s, 1 H), 6.58 (s, 1 H), 6.48 (s, 2 H), 6.25 (s, 2 H). MS (LC/MS) R.T. = 1.59; [M+H]⁺ = 213.0。 Step B: 6,7-Dichloroisoquinolin-3-amine and 5,6-Dichloroisoquinolin-3-amine

To sulfuric acid (4 ml, 75 mmol), N- (3,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide (2 g, 6.6 mmol) was added at room temperature. The reaction was heated at 40 ° C. for 49 hours. TLC and LC / MS indicated the presence of product. The reaction was cooled to room temperature and quenched with aqueous NaOH (about 15M) until the reaction mixture was pH ˜7. The crude product was extracted with ethyl acetate (2 × 50 ml) and the organics were dried over MgSO ₄ , filtered and concentrated in vacuo to give the product. The crude product was purified by chromatography (Biotage, 100% ethyl acetate to [90/10% ethyl acetate / MeOH]) and the regioisomers 6,7-dichloroisoquinolin-1-amine and 5,6-dichloroisoquinoline- An approximately 1: 1 mixture of 3-amine was obtained as a dark yellow powder (1.2 g, 5.64 mmol, 86.0% yield). The regioisomer was used without separation.
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 8.90 (s, 1 H), 8.83 (s, 1 H), 8.12 (s, 1 H), 7.89 (s, 1 H), 7.84 (d, J = 8.54 Hz, 1 H), 7.28 (d, J = 8.85 Hz, 1 H), 6.83 (s, 1 H), 6.58 (s, 1 H), 6.48 (s, 2 H), 6.25 (s, 2 H). MS (LC / MS) RT = 1.59; [M + H] ⁺ = 213.0.

ジクロロメタン(２０ml)中の６,７−ジクロロイソキノリン−３−アミンおよび５,６−ジクロロイソキノリン−３−アミン(０.４１０ｇ, １.９２４mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.４６９ｇ, ２.０２１mmol)を加え、反応混合物を４０℃で４時間撹拌した。反応物を室温まで冷却し、クロマトグラフィーを行い(Biotage, １０〜１００％ 酢酸エチル／ヘキサン)、分離した位置異性体６,７−ジクロロ−３−イソチオシアナトイソキノリン(０.２ｇ, ０.７８４mmol, 収率４０.７％)および５,６−ジクロロ−３−イソチオシアナトイソキノリンを黄色の固体として得た(０.２３ｇ, ０.９０２mmol, 収率４６.８％)。
５,６−ジクロロ−３−イソチオシアナトイソキノリン： ¹H NMR (500 MHz, CDCl₃) δ ppm 9.10 (s, 1 H), 7.87 (d, J=8.85 Hz, 1 H), 7.82 (s, 1 H), 7.66 (d, J=8.55 Hz, 1 H). MS (LC/MS) R.T. = 3.63; [M+H]⁺ = 255.0。
６,７−ジクロロ−３−イソチオシアナトイソキノリン： ¹H NMR (500 MHz, CDCl₃) δ ppm 9.04 (s, 1 H), 8.11 (s, 1 H), 7.94 (s, 1 H), 7.37 (s, 1 H). MS (LC/MS) R.T. = 3.42; [M+H]⁺ = 255.0。 Step C: 6,7-dichloro-3-isothiocyanatoisoquinoline and 5,6-dichloro-3-isothiocyanatoisoquinoline

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′-thiocarbonyldipyridine-2 ( 1H) -one (0.469 g, 2.021 mmol) was added and the reaction mixture was stirred at 40 ° C. for 4 h. The reaction was cooled to room temperature and chromatographed (Biotage, 10-100% ethyl acetate / hexanes), and the regioisomer 6,7-dichloro-3-isothiocyanatoisoquinoline (0.2 g, 0.784 mmol) isolated. , Yield 40.7%) and 5,6-dichloro-3-isothiocyanatoisoquinoline as a yellow solid (0.23 g, 0.902 mmol, 46.8% yield).
5,6-dichloro-3-isothiocyanatoisoquinoline: ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.10 (s, 1 H), 7.87 (d, J = 8.85 Hz, 1 H), 7.82 (s, 1 H), 7.66 (d, J = 8.55 Hz, 1 H). MS (LC / MS) RT = 3.63; [M + H] ⁺ = 255.0.
6,7-dichloro-3-isothiocyanatoisoquinoline: ¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.04 (s, 1 H), 8.11 (s, 1 H), 7.94 (s, 1 H), 7.37 (s, 1 H). MS (LC / MS) RT = 3.42; [M + H] ⁺ = 255.0.

ＤＭＦ(１０ml)中の６,７−ジクロロ−３−イソチオシアナトイソキノリン(０.１３ｇ, ０.５１０mmol)に、炭酸セシウム(０.４２ｇ, １.２７mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１１８ｇ, ０.５１５mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物を、ＤＭＦ(１０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.２３８ml, １.５２９mmol)で処理した。反応物を９０℃で１８時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。これをクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(６,７−ジクロロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(０.１２ｇ, ０.３１２mmol, 収率６１.２％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.03 (s, 1 H), 8.87 (s, 1 H), 7.96 (s, 1 H), 7.83 (s, 1 H), 7.24 (s, 1 H), 3.88 - 4.06 (m, 1 H), 3.60 - 3.74 (m, 1 H), 3.42 (d, J=14.65 Hz, 1 H), 2.82 - 3.15 (m, 5 H), 2.23 - 2.34 (m, 1 H), 2.18 (s, 1 H), 1.72 - 1.87 (m, 1 H), 1.48 - 1.70 (m, 2 H). MS (LC/MS) R.T. = 1.63; [M+H]⁺ = 377.1。 Step D: (R) -N- (6,7-Dichloroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

6,7-Dichloro-3-isothiocyanatoisoquinoline (0.13 g, 0.510 mmol) in DMF (10 ml) was added to cesium carbonate (0.42 g, 1.27 mmol) and (S) -3- (aminomethyl). Quinuclidin-3-ol dihydrochloride (0.118 g, 0.515 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with DMF (10 ml) and N, N′-diisopropylcarbodiimide (0.238 ml, 1.529 mmol). The reaction was heated at 90 ° C. for 18 hours. The reaction was cooled to room temperature and concentrated to give the crude product. This was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -N- (6,7-dichloroisoquinolin-3-yl) -4H-1′- Azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (0.12 g, 0.312 mmol, yield 61.2%).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.03 (s, 1 H), 8.87 (s, 1 H), 7.96 (s, 1 H), 7.83 (s, 1 H), 7.24 (s, 1 H ), 3.88-4.06 (m, 1 H), 3.60-3.74 (m, 1 H), 3.42 (d, J = 14.65 Hz, 1 H), 2.82-3.15 (m, 5 H), 2.23-2.34 (m , 1 H), 2.18 (s, 1 H), 1.72-1.87 (m, 1 H), 1.48-1.70 (m, 2 H). MS (LC / MS) RT = 1.63; [M + H] ⁺ = 377.1.

ＤＭＦ(１０ml)中の５,６−ジクロロ−３−イソチオシアナトイソキノリン(０.１１ｇ, ０.４３１mmol)に、炭酸セシウム(０.３５１ｇ, １.０７８mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１００ｇ, ０.４３５mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製した(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)。生成物をＤＭＦ(１０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.２０２ml, １.２９３mmol)で処理した。反応物を９０℃で１８時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、(Ｒ)−Ｎ−(５,６−ジクロロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の粉末として得た(０.０８４ｇ, ０.２１８mmol, 収率５０.６％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.09 (s, 1 H), 8.93 (s, 1 H), 7.63 - 7.82 (m, 2 H), 7.40 (d, J=8.55 Hz, 1 H), 3.99 (d, J=9.16 Hz, 1 H), 3.78 (d, J=8.85 Hz, 1 H), 3.51 (d, J=14.65 Hz, 1 H), 3.30 (d, J=14.65 Hz, 1 H), 2.90 - 3.23 (m, 4 H), 2.33 - 2.48 (m, 1 H), 2.29 (s, 1 H), 1.83 - 1.94 (m, 1 H), 1.62 - 1.83 (m, J=42.12 Hz, 2 H). MS (LC/MS) R.T. = 1.57; [M+H]⁺ = 377.1。 Example 281
(R) -N- (5,6-Dichloroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

5,6-Dichloro-3-isothiocyanatoisoquinoline (0.11 g, 0.431 mmol) in DMF (10 ml) was added to cesium carbonate (0.351 g, 1.078 mmol) and (S) -3- (aminomethyl). Quinuclidin-3-ol dihydrochloride (0.100 g, 0.435 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH). The product was treated with DMF (10 ml) and N, N′-diisopropylcarbodiimide (0.202 ml, 1.293 mmol). The reaction was heated at 90 ° C. for 18 hours. The reaction was cooled to room temperature and concentrated to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH), (R) -N- (5,6-dichloroisoquinolin-3-yl) -4H-1 '-Azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow powder (0.084 g, 0.218 mmol, 50.6% yield).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.09 (s, 1 H), 8.93 (s, 1 H), 7.63-7.82 (m, 2 H), 7.40 (d, J = 8.55 Hz, 1 H) , 3.99 (d, J = 9.16 Hz, 1 H), 3.78 (d, J = 8.85 Hz, 1 H), 3.51 (d, J = 14.65 Hz, 1 H), 3.30 (d, J = 14.65 Hz, 1 H), 2.90-3.23 (m, 4 H), 2.33-2.48 (m, 1 H), 2.29 (s, 1 H), 1.83-1.94 (m, 1 H), 1.62-1.83 (m, J = 42.12 Hz, 2 H). MS (LC / MS) RT = 1.57; [M + H] ⁺ = 377.1.

(Ｒ)−Ｎ−(３,４−ジクロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の方法によって、２−アミノ−３,４−ジクロロピリジンから出発して製造した。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 9.10 (s, 1 H), 8.08 (d, J=5.49 Hz, 1 H), 7.13 (d, J=5.49 Hz, 1 H), 3.86 (d, J=9.77 Hz, 1 H), 3.60 (d, J=9.77 Hz, 1 H), 2.96 - 3.05 (m, 2 H), 2.77 (t, J=7.63 Hz, 2 H), 2.66 (t, J=7.78 Hz, 2 H), 1.97 - 2.05 (m, 1 H), 1.86 - 1.94 (m, 1 H), 1.54 - 1.63 (m, 2 H), 1.43 - 1.51 (m, 1 H). MS (LC/MS) R.T. = 0.78; [M+H]⁺ = 327.0。 Example 282
(R) -N- (3,4-Dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (3,4-Dichloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was carried out. Prepared by the method of Example 267 starting from 2-amino-3,4-dichloropyridine.
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 9.10 (s, 1 H), 8.08 (d, J = 5.49 Hz, 1 H), 7.13 (d, J = 5.49 Hz, 1 H), 3.86 ( d, J = 9.77 Hz, 1 H), 3.60 (d, J = 9.77 Hz, 1 H), 2.96-3.05 (m, 2 H), 2.77 (t, J = 7.63 Hz, 2 H), 2.66 (t , J = 7.78 Hz, 2 H), 1.97-2.05 (m, 1 H), 1.86-1.94 (m, 1 H), 1.54-1.63 (m, 2 H), 1.43-1.51 (m, 1 H). MS (LC / MS) RT = 0.78; [M + H] ⁺ = 327.0.

(Ｒ)−Ｎ−(３−クロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の方法によって、２−アミノ−３−クロロピリジンから出発して製造した。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 9.06 (s, 1 H), 8.14 - 8.19 (m, 1 H), 7.74 - 7.79 (m, J=7.78, 1.83, 1.83, 1.68 Hz, 1 H), 6.86 - 6.91 (m, 1 H), 3.81 - 3.89 (m, 1 H), 3.55 - 3.63 (m, 1 H), 2.96 - 3.04 (m, 2 H), 2.78 (t, J=7.63 Hz, 2 H), 2.67 (t, J=7.63 Hz, 2 H), 1.96 - 2.02 (m, 1 H), 1.86 - 1.92 (m, J=5.65, 3.20 Hz, 1 H), 1.54 - 1.63 (m, J=6.87, 3.66, 3.51 Hz, 2 H), 1.42 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 0.26; [M+H]⁺ = 293.0。 Example 283
(R) -N- (3-chloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (3-Chloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 267. Prepared starting from 2-amino-3-chloropyridine.
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 9.06 (s, 1 H), 8.14-8.19 (m, 1 H), 7.74-7.79 (m, J = 7.78, 1.83, 1.83, 1.68 Hz, 1 H), 6.86-6.91 (m, 1 H), 3.81-3.89 (m, 1 H), 3.55-3.63 (m, 1 H), 2.96-3.04 (m, 2 H), 2.78 (t, J = 7.63 Hz, 2 H), 2.67 (t, J = 7.63 Hz, 2 H), 1.96-2.02 (m, 1 H), 1.86-1.92 (m, J = 5.65, 3.20 Hz, 1 H), 1.54-1.63 ( m, J = 6.87, 3.66, 3.51 Hz, 2 H), 1.42-1.49 (m, 1 H). MS (LC / MS) RT = 0.26; [M + H] ⁺ = 293.0.

(Ｒ)−Ｎ−(５−クロロ−３−フルオロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の方法によって、２−アミノ−３−フルオロ−５−クロロピリジンから出発して製造した。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 8.81 (s, 1 H), 8.05 (s, 1 H), 7.79 (d, J=10.07 Hz, 1 H), 3.83 (d, J=9.46 Hz, 1 H), 3.58 (d, J=9.46 Hz, 1 H), 2.99 (s, 2 H), 2.71 - 2.80 (m, 2 H), 2.61 - 2.70 (m, 2 H), 2.00 (s, 1 H), 1.83 - 1.92 (m, 1 H), 1.53 - 1.62 (m, 2 H), 1.41 - 1.50 (m, 1 H). MS (LC/MS) R.T. = 0.52; [M+]⁺ = 311.0。 Example 284
(R) -N- (5-Chloro-3-fluoropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Chloro-3-fluoropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 267 starting from 2-amino-3-fluoro-5-chloropyridine.
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 8.81 (s, 1 H), 8.05 (s, 1 H), 7.79 (d, J = 10.07 Hz, 1 H), 3.83 (d, J = 9.46 Hz, 1 H), 3.58 (d, J = 9.46 Hz, 1 H), 2.99 (s, 2 H), 2.71-2.80 (m, 2 H), 2.61-2.70 (m, 2 H), 2.00 (s , 1 H), 1.83-1.92 (m, 1 H), 1.53-1.62 (m, 2 H), 1.41-1.50 (m, 1 H). MS (LC / MS) RT = 0.52; [M +] ⁺ = 311.0.

(Ｒ)−Ｎ−(６−クロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の方法によって、２−アミノ−６−クロロピリジンから出発して製造した。
¹H NMR (500 MHz, DMSO-D₆) δ ppm 8.33 - 8.42 (m, 1 H), 7.60 - 7.68 (m, 1 H), 6.94 (d, J=7.02 Hz, 1 H), 6.72 - 6.81 (m, 1 H), 3.86 (d, J=9.46 Hz, 1 H), 3.57 (d, J=10.07 Hz, 1 H), 2.97 (s, 2 H), 2.69 - 2.78 (m, 2 H), 2.63 - 2.68 (m, J=7.63, 7.63 Hz, 2 H), 1.95 - 2.03 (m, 1 H), 1.83 - 1.92 (m, 1 H), 1.53 - 1.62 (m, 2 H), 1.41 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 0.43; [M+H]⁺ = 293.0。 Example 285
(R) -N- (6-Chloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Chloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 267. Prepared starting from 2-amino-6-chloropyridine.
¹ H NMR (500 MHz, DMSO-D ₆ ) δ ppm 8.33-8.42 (m, 1 H), 7.60-7.68 (m, 1 H), 6.94 (d, J = 7.02 Hz, 1 H), 6.72-6.81 (m, 1 H), 3.86 (d, J = 9.46 Hz, 1 H), 3.57 (d, J = 10.07 Hz, 1 H), 2.97 (s, 2 H), 2.69-2.78 (m, 2 H) , 2.63-2.68 (m, J = 7.63, 7.63 Hz, 2 H), 1.95-2.03 (m, 1 H), 1.83-1.92 (m, 1 H), 1.53-1.62 (m, 2 H), 1.41- 1.49 (m, 1 H). MS (LC / MS) RT = 0.43; [M + H] ⁺ = 293.0.

(Ｒ)−Ｎ−(４,６−ジクロロピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の方法によって、２−アミノ−４,６−ジクロロピリジンから出発して製造した。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.43 (s, 1 H), 7.13 (s, 1 H), 6.84 (s, 1 H), 3.86 (d, J=9.77 Hz, 1 H), 3.59 (d, J=10.07 Hz, 1 H), 2.98 (s, 2 H), 2.58 - 2.86 (m, 4 H), 1.94 - 2.13 (m, 1 H), 1.78 - 1.95 (m, 1 H), 1.36 - 1.65 (m, 3 H). MS (LC/MS) R.T. = 0.87; [M+H]⁺ = 327.0。 Example 286
(R) -N- (4,6-dichloropyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4,6-dichloropyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was carried out. Prepared by the method of Example 267 starting from 2-amino-4,6-dichloropyridine.
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.43 (s, 1 H), 7.13 (s, 1 H), 6.84 (s, 1 H), 3.86 (d, J = 9.77 Hz, 1 H), 3.59 (d, J = 10.07 Hz, 1 H), 2.98 (s, 2 H), 2.58-2.86 (m, 4 H), 1.94-2.13 (m, 1 H), 1.78-1.95 (m, 1 H) , 1.36-1.65 (m, 3 H). MS (LC / MS) RT = 0.87; [M + H] ⁺ = 327.0.

Example 287
(R) -N- (2-methoxy-3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine

ＤＭＦ(２５ml)中の、４−ブロモピリジン−２−アミン(０.５ｇ, ２.８mmol)、２−メトキシピリジン−３−イルボロン酸(０.５２ｇ, ３.４mmol)に、１Ｎ 炭酸ナトリウム(１０ml, ２.３mmol)を、次に塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体(０.２１ｇ, ０.２６mmol)を加えた。反応混合物を８５℃で３時間撹拌し、室温まで冷却した。生成物を酢酸エチル(２×５０ml)で抽出し、ＭｇＳＯ_４で乾燥させ、濾過し、真空で濃縮した。粗生成物をクロマトグラフィーによって精製し(Biotage, １００から９０／１０％ 酢酸エチル−酢酸エチル／メタノール)、２−メトキシ−３,４'−ビピリジン−２'−アミンを褐色の粉末として得た(０.５３ｇ, ２.６３mmol, 収率９３％)。生成物を次の工程に直接用いた。 Step A: N- (2,4-dichlorobenzyl) -2,2-diethoxyacetimidoamide

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 to 1N sodium carbonate (10 ml). , 2.3 mmol) followed by 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex (0.21 g, 0.26 mmol). The reaction mixture was stirred at 85 ° C. for 3 hours and cooled to room temperature. The product was extracted with ethyl acetate (2 × 50 ml), dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (Biotage, 100 to 90/10% ethyl acetate-ethyl acetate / methanol) to give 2-methoxy-3,4'-bipyridin-2'-amine as a brown powder ( 0.53 g, 2.63 mmol, 93% yield). The product was used directly in the next step.

ジクロロメタン(２０ml)中の２−メトキシ−３,４'−ビピリジン−２'−アミン(０.５３ｇ, ２.６３mmol)に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.６２ｇ, ２.７mmol)を加え、反応混合物を４０℃で４時間撹拌した。反応物を室温まで冷却し、クロマトグラフィーを行い(Biotage, １０〜１００％ 酢酸エチル／ヘキサン)、２'−イソチオシアナト−２−メトキシ−３,４'−ビピリジンを得た(０.４６ｇ, １.９mmol, 収率７１.８％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 8.67 (d, J=2.44 Hz, 1 H), 8.26 (dd, J=4.88, 1.53 Hz, 1 H), 8.15 (dd, J=8.24, 2.44 Hz, 1 H), 7.89 (dd, J=7.32, 1.53 Hz, 1 H), 7.48 (d, J=8.24 Hz, 1 H), 7.15 (dd, J=7.32, 4.88 Hz, 1 H), 3.91 (s, 3 H). MS (LC/MS) R.T. = 2.87; [M+H]⁺ = 244.9。 Step B: 2'-isothiocyanato-2-methoxy-3,4'-bipyridine

2-Methoxy-3,4'-bipyridin-2′-amine (0.53 g, 2.63 mmol) in dichloromethane (20 ml) was added to 1,1′-thiocarbonyldipyridin-2 (1H) -one (0 .62 g, 2.7 mmol) was added and the reaction mixture was stirred at 40 ° C. for 4 hours. The reaction was cooled to room temperature and chromatographed (Biotage, 10-100% ethyl acetate / hexane) to give 2′-isothiocyanato-2-methoxy-3,4′-bipyridine (0.46 g, 1. 9 mmol, yield 71.8%).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 8.67 (d, J = 2.44 Hz, 1 H), 8.26 (dd, J = 4.88, 1.53 Hz, 1 H), 8.15 (dd, J = 8.24, 2.44 Hz, 1 H), 7.89 (dd, J = 7.32, 1.53 Hz, 1 H), 7.48 (d, J = 8.24 Hz, 1 H), 7.15 (dd, J = 7.32, 4.88 Hz, 1 H), 3.91 (s, 3 H). MS (LC / MS) RT = 2.87; [M + H] ⁺ = 244.9.

ＤＭＦ(２０ml)中の２'−イソチオシアナト−２−メトキシ−３,４'−ビピリジン(０.０９ｇ, ０.３７mmol)に、Ｅｔ_３Ｎ(０.１１ml, ０.８１mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.０９ｇ, ０.３７mmol)を室温で加えた。反応物を７０℃で２時間撹拌した。反応物を室温まで冷却し、真空で濃縮した。粗製のウレアをクロマトグラフィーによって精製し(biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、純粋なウレア中間体を得た。生成物をＤＭＦ(２０ml)およびＮ,Ｎ'−ジイソプロピルカルボジイミド(０.１７ml, １.１mmol)で処理した。反応物を９０℃で１８時間加熱した。反応物を室温まで冷却し、濃縮し、粗生成物を得た。粗生成物をクロマトグラフィーによって精製し(Biotage, ８５％ ＣＨＣｌ_３, １４％ ＭｅＯＨ, １％ ＮＨ_４ＯＨ)、生成物を含むフラクションを合わせた。ＬＣ／ＭＳおよび^１Ｈ−ＮＭＲにより、幾らかの不純物が存在するかもしれないことが示された。純粋でない生成物について逆相ＨＰＬＣを行い、(Ｒ)−Ｎ−(２−メトキシ−３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(０.０２ｇ, ０.０５mmol, 収率１４.０５％)。
¹H NMR (500 MHz, DMSO-D6) δ ppm 9.07 (s, 1 H), 8.43 (s, 1 H), 8.17 (dd, J=4.73, 1.68 Hz, 1 H), 7.79 (d, J=7.32 Hz, 2 H), 7.10 (dd, J=7.32, 4.88 Hz, 1 H), 6.79 - 6.92 (m, 1 H), 3.76 - 3.97 (m, 4 H), 3.51 - 3.66 (m, 1 H), 2.92 - 3.09 (m, 2 H), 2.59 - 2.82 (m, 4 H), 1.85 - 2.03 (m, 2 H), 1.53 - 1.71 (m, 2 H), 1.35 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 1.05; [M+H]⁺ = 366.1。 Step C: (R) -N- (2-methoxy-3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane ] -2-Amine

2'-isothiocyanato-2-methoxy-3,4'-bipyridine (0.09 g, 0.37 mmol) in DMF (20 ml) was added to Et ₃ N (0.11 ml, 0.81 mmol) and (S) -3. -(Aminomethyl) quinuclidin-3-ol dihydrochloride (0.09 g, 0.37 mmol) was added at room temperature. The reaction was stirred at 70 ° 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 ₃ , 14% MeOH, 1% NH ₄ OH) to give the pure urea intermediate. The product was treated with DMF (20 ml) and N, N′-diisopropylcarbodiimide (0.17 ml, 1.1 mmol). The reaction was heated at 90 ° C. for 18 hours. The reaction was cooled to room temperature and concentrated to give the crude product. The crude product was purified by chromatography (Biotage, 85% CHCl ₃ , 14% MeOH, 1% NH ₄ OH) and the fractions containing the product were combined. LC / MS and ¹ H-NMR indicated that some impurities may be present. The impure product was subjected to reverse phase HPLC and (R) -N- (2-methoxy-3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] Octane] -2-amine was obtained as a white powder (0.02 g, 0.05 mmol, 14.05% yield).
¹ H NMR (500 MHz, DMSO-D6) δ ppm 9.07 (s, 1 H), 8.43 (s, 1 H), 8.17 (dd, J = 4.73, 1.68 Hz, 1 H), 7.79 (d, J = 7.32 Hz, 2 H), 7.10 (dd, J = 7.32, 4.88 Hz, 1 H), 6.79-6.92 (m, 1 H), 3.76-3.97 (m, 4 H), 3.51-3.66 (m, 1 H ), 2.92-3.09 (m, 2 H), 2.59-2.82 (m, 4 H), 1.85-2.03 (m, 2 H), 1.53-1.71 (m, 2 H), 1.35-1.49 (m, 1 H ). MS (LC / MS) RT = 1.05; [M + H] ⁺ = 366.1.

Example 288
(R) -N- (Benzo [d] oxazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

オーブン乾燥した丸底フラスコに、ジ(１Ｈ−イミダゾール−１−イル)メタンイミン (５００mg, ３.１０mmol)、２−アミノフェノール(１８８mg, １.７２４mmol)および無水ＴＨＦ(２０ml)を室温で入れた。得られた懸濁液をＮ_２下で２時間還流し、ＬＣ／ＭＳに基づいて変換の完了を見た。溶媒を真空で除去し、残渣を、Biotage Flash Collectorで精製し、３０〜８０％ ＥｔＯＡｃ／ヘキサン(１２００ml)で溶出し、予測された生成物であるベンゾ[ｄ]オキサゾール−２−アミンを白色の固体として得た(２００mg, １.５mmol, 収率８７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 6.20 (br. s., 2 H) 7.02 - 7.11 (m, 1 H) 7.17 - 7.22 (m, 1 H) 7.29 (d, J=7.53 Hz, 1 H) 7.36 (d, J=7.03 Hz, 1 H ). MS (LC/MS) R.T. = 1.05; [M+H]⁺ = 134.96。 Step A: Benzo [d] oxazol-2-amine

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 for 2 h under N ₂ and saw complete conversion based on LC / MS. The solvent is removed in vacuo and the residue is purified on a Biotage Flash Collector, eluting with 30-80% EtOAc / hexane (1200 ml) to give the expected product, benzo [d] oxazol-2-amine, as white. Obtained as a solid (200 mg, 1.5 mmol, 87% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 6.20 (br. S., 2 H) 7.02-7.11 (m, 1 H) 7.17-7.22 (m, 1 H) 7.29 (d, J = 7.53 Hz, 1 H) 7.36 (d, J = 7.03 Hz, 1 H). MS (LC / MS) RT = 1.05; [M + H] ⁺ = 134.96.

ＤＭＦ(１０ml)中のベンゾ[ｄ]オキサゾール−２−アミン(２００mg, １.４９１mmol)の無色の溶液に、水酸化ナトリウム(２０Ｎ, １４９μｌ, ２.９８mmol)を加え、緑色の懸濁液を得た。混合物を室温で１５分間撹拌した。二硫化炭素(２２５μｌ, ３.７３mmol)を加え、暗褐色の溶液を得た。反応物を室温で１５分間撹拌し、水酸化ナトリウム(２０Ｎ, １４９μｌ, ２.９８mmol)を添加し、さらに１０分間撹拌した。ヨードメタン(２２４μｌ, ３.５８mmol)を滴下した。１２分後に緑色の固体が沈殿した。反応物をさらに２時間撹拌した。固体を濾過によって集め、ＤＭＦ(２×１ml)で、そしてＨ_２Ｏ(２×１ml)で洗浄し、ハウスバキューム(house vacuum)下で３０分間乾燥させ、さらに、オーブン中で、真空で一夜乾燥させ、予測された生成物であるベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチルを白色の固体として得た(２５８.５mg, １.０８５mmol, 収率７２.７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 2.70 (s, 6 H) 7.24 - 7.34 (m, 2 H) 7.45 - 7.50 (m, 1 H) 7.66 - 7.74 (m, 1 H). MS (LC/MS) R.T. = 1.76, [M+H]⁺ = 238.96。 Step B: Benzo [d] oxazol-2-ylcarbonimidodithioate dimethyl

To a colorless solution of benzo [d] oxazol-2-amine (200 mg, 1.491 mmol) in DMF (10 ml), sodium hydroxide (20N, 149 μl, 2.98 mmol) is added to give a green suspension. It was. The mixture was stirred at room temperature for 15 minutes. Carbon disulfide (225 μl, 3.73 mmol) was added to give a dark brown solution. The reaction was stirred at room temperature for 15 minutes, sodium hydroxide (20N, 149 μl, 2.98 mmol) was added and stirred for an additional 10 minutes. Iodomethane (224 μl, 3.58 mmol) was added dropwise. A green solid precipitated after 12 minutes. The reaction was stirred for an additional 2 hours. The solid was collected by filtration, washed with DMF (2 × 1 ml) and with H ₂ O (2 × 1 ml), dried under house vacuum for 30 minutes and further dried in an oven overnight in vacuo. The expected product, dimethyl benzo [d] oxazol-2-ylcarbonimidodithioate, was obtained as a white solid (258.5 mg, 1.085 mmol, yield 72.7%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 2.70 (s, 6 H) 7.24-7.34 (m, 2 H) 7.45-7.50 (m, 1 H) 7.66-7.74 (m, 1 H). MS (LC / MS) RT = 1.76, [M + H] ⁺ = 238.96.

Step C: (R) -N- (Benzo [d] oxazol-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine 10 ml Into a vial of (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (69.5 mg, 0.361 mmol), DMF (2 ml), DIEA (0.063 ml, 0.361 mmol) and Cs ₂ CO ₃ (235 mg, 0.722 mmol) was charged 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 hour. LC / MS showed that the starting material was consumed. The mixture was diluted with MeOH and purified by preparative HPLC, and the expected product (R) -N- (benzo [d] oxazol-2-yl) -4H-1′-azaspiro [oxazole-5, 3'-Bicyclo [2.2.2] octane] -2-amine was obtained as a tan gum (101.5 mg, 0.323 mmol, 90% yield).
¹ H NMR (400 MHz, acetone-d ₆ ) δ ppm 2.07-2.14 (m, 2 H) 2.20 (ddd, J = 8.78, 5.27, 3.26 Hz, 2 H) 2.33-2.45 (m, 1 H) 2.62 ( d, J = 2.26 Hz, 1 H) 3.34-3.47 (m, 3 H) 3.48-3.58 (m, 1 H) 3.75-3.88 (m, 2 H) 4.15 (d, J = 10.54 Hz, 1 H) 4.32 (d, J = 10.54 Hz, 1 H) 7.13-7.26 (m, 2 H) 7.41 (td, J = 3.70, 1.63 Hz, 1 H) 9.24 (br, s, 1H). MS (LC / MS) RT = 0.792, [M + H] ⁺ = 299.17.

Example 289
(R) -N- (5-Chlorobenzo [d] oxazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

ＤＭＦ(５ml)中の５−クロロベンゾ[ｄ]オキサゾール−２−アミン(７００mg, ４.１５mmol)の褐色の溶液に、水酸化ナトリウム(２０Ｎ, ４１５μｌ, ８.３０mmol)を加え、灰色の懸濁液を得た。混合物を室温で１５分間撹拌した。二硫化炭素(６２６μｌ, １０.３８mmol)を室温で加え、褐色の溶液を得た。混合物を室温で１５分間撹拌し、水酸化ナトリウム(２０Ｎ, ２０８μｌ, ４.１６mmol)を加えた。１０分後、ヨードメタン(６２３μｌ, ９.９７mmol)を滴下した。溶液から灰色の固体が生じた。反応物をさらに室温で２時間撹拌した。固体を濾過によって集め、ＤＭＦ／Ｈ_２Ｏ(５０：５０, ２×２ml)で洗浄し、ハウスバキューム下で３０分間乾燥させ、さらに、オーブン中、真空下で６５℃で１.５時間乾燥させ、予測された生成物である５−クロロベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチルを、次の工程に用いるのに十分な程純粋な灰白色の固体として得た(７８０mg, ２.８６mmol, 収率６８.９％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.69 (s, 6 H) 7.37 (dd, J=8.53, 2.26 Hz, 1 H) 7.66 (d, J=9.03 Hz, 1 H) 7.75 (d, J=1.76 Hz, 1 H). MS (LC/MS) R.T. = 1.44; [M+H]⁺ = 272.9。 Step A: Dimethyl 5-chlorobenzo [d] oxazol-2-ylcarbonimidodithioate

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 μl, 8.30 mmol) and a gray suspension. Got. The mixture was stirred at room temperature for 15 minutes. Carbon disulfide (626 μl, 10.38 mmol) was added at room temperature to give a brown solution. The mixture was stirred at room temperature for 15 minutes and sodium hydroxide (20N, 208 μl, 4.16 mmol) was added. After 10 minutes, iodomethane (623 μl, 9.97 mmol) was added dropwise. A gray solid resulted from the solution. The reaction was further stirred at room temperature for 2 hours. The solid was collected by filtration, washed with DMF / H ₂ O (50:50, 2 × 2 ml), dried under house vacuum for 30 minutes, and further dried in an oven at 65 ° C. under vacuum for 1.5 hours. The expected product, dimethyl 5-chlorobenzo [d] oxazol-2-ylcarbonimidodithioate, was obtained as an off-white solid (780 mg, 2.86 mmol) sufficient to be used in the next step. Yield 68.9%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 2.69 (s, 6 H) 7.37 (dd, J = 8.53, 2.26 Hz, 1 H) 7.66 (d, J = 9.03 Hz, 1 H) 7.75 (d , J = 1.76 Hz, 1 H). MS (LC / MS) RT = 1.44; [M + H] ⁺ = 272.9.

１０mlのバイアルに、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１０６mg, ０.５５０mmol)、ＤＭＦ(２ml)、ＤＩＥＡ(０.０９６ml, ０.５５０mmol)およびＣｓ_２ＣＯ_３(３５８mg, １.１００mmol)を室温で入れ、次に５−クロロベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチル(１５０mg, ０.５５０mmol)を入れた。得られた懸濁液を室温で１時間撹拌した。ＬＣ／ＭＳにより、出発物質が消費されたことが示された。反応混合物をＭｅＯＨで希釈し、分取ＨＰＬＣによって精製し、予測された生成物である(Ｒ)−Ｎ−(５−クロロベンゾ[ｄ]オキサゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の固体として得た(１０６.５mg, ０.３mmol, 収率５３.０％)。
¹H NMR (400 MHz, アセトン-d₆) δ ppm 2.08 - 2.30 (m, 3 H) 2.35 - 2.48 (m, 1 H) 2.60 - 2.73 (m, 1 H) 3.48 (qd, J=7.53, 7.28 Hz, 3 H) 3.54 - 3.68 (m, 1 H) 3.79 - 4.00 (m, 2 H) 4.18 (d, J=10.54 Hz, 1 H) 4.35 (d, J=10.54 Hz, 1 H) 7.19 (dd, J=8.53, 2.01 Hz, 1 H) 7.30 - 7.47 (m, 2 H) 9.10 (br. s., 1 H). MS(LC/MS) R.T. = 1.56; [M+H]⁺ = 333.13。 Step B: (R) -N- (5-Chlorobenzo [d] oxazol-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

In a 10 ml vial (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (106 mg, 0.550 mmol), DMF (2 ml), DIEA (0.096 ml, 0.550 mmol) and Cs ₂ CO ₃ (358 mg, 1.100 mmol) was charged 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 hour. LC / MS showed that the starting material was consumed. The reaction mixture was diluted with MeOH and purified by preparative HPLC and the expected product (R) -N- (5-chlorobenzo [d] oxazol-2-yl) -4H-1′-azaspiro [oxazole -5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a white solid (106.5 mg, 0.3 mmol, 53.0% yield).
¹ H NMR (400 MHz, acetone-d ₆ ) δ ppm 2.08-2.30 (m, 3 H) 2.35-2.48 (m, 1 H) 2.60-2.73 (m, 1 H) 3.48 (qd, J = 7.53, 7.28 Hz, 3 H) 3.54-3.68 (m, 1 H) 3.79-4.00 (m, 2 H) 4.18 (d, J = 10.54 Hz, 1 H) 4.35 (d, J = 10.54 Hz, 1 H) 7.19 (dd , J = 8.53, 2.01 Hz, 1 H) 7.30-7.47 (m, 2 H) 9.10 (br. S., 1 H). MS (LC / MS) RT = 1.56; [M + H] ⁺ = 333.13.

Example 290
(R) -N- (Oxazolo [4,5-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

オーブン乾燥した丸底フラスコに、ジ(１Ｈ−イミダゾール−１−イル)メタンイミン(５００mg, ３.１０mmol)、２−アミノピリジン−３−オール(１７１mg, １.５５１mmol)および無水ＴＨＦ(２０ml)を室温で入れた。得られた懸濁液を、Ｎ_２下、１時間還流した。ＬＣ／ＭＳにより、出発物質が消費されたことが示された。溶媒を真空で除去し、残渣をさらに精製することなく次の工程に用いた。
MS(LC/MS) R.T. = 0.235; [M+H]⁺ = 136.09。 Step A: Oxazolo [4,5-b] pyridin-2-amine

To an oven-dried round bottom flask was added 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. I put it in. The resulting suspension, N ₂ under, and refluxed for 1 hour. LC / MS showed that the starting material was consumed. The solvent was removed in vacuo and the residue was used in the next step without further purification.
MS (LC / MS) RT = 0.235; [M + H] ⁺ = 136.09.

ＤＭＦ(１２ml)中の粗製のオキサゾロ[４,５−ｂ]ピリジン−２−アミン(８１１mg, ６mmol)(工程Ａより)に、ＮａＯＨ(２０Ｎ, ６００μｌ, １２.００mmol)を加え、黄褐色の溶液を得た。これを室温で１５分間撹拌した。二硫化炭素(９０４μｌ, １５.００mmol)を加え、橙色の溶液を得た。混合物を室温で１５分間撹拌し、ＮａＯＨ(２０Ｎ, ６００μｌ, １２.００mmol)を加え、１０分間撹拌を続け、暗赤色の溶液を得た。ヨードメタン(９００μｌ, １４.４０mmol)を滴下し、１時間後に黄色の固体の沈殿が起こり、約８０％が変換された。混合物をＭｅＯＨで希釈し、分取ＨＰＬＣによって精製し、予測された生成物であるオキサゾロ[４,５−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルを白色の固体として得た(３５mg, ０.１４６mmol, 収率２.４％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 2.71 (s, 6 H) 7.22 (dd, J=8.03, 5.02 Hz, 1 H) 7.72 (dd, J=8.03, 1.25 Hz, 1 H) 8.49 (dd, J=5.02, 1.51 Hz, 1 H). MS (LC/MS) R.T. = 1.358; [M+H] = 240.04。 Step B: Dimethyl oxazolo [4,5-b] pyridin-2-ylcarbonimidodithioate

To the crude oxazolo [4,5-b] pyridin-2-amine (811 mg, 6 mmol) (from step A) in DMF (12 ml) was added NaOH (20N, 600 μl, 12.00 mmol) to give a tan solution Got. This was stirred at room temperature for 15 minutes. Carbon disulfide (904 μl, 15.00 mmol) was added to give an orange solution. The mixture was stirred at room temperature for 15 minutes, NaOH (20N, 600 μl, 12.00 mmol) was added and stirring was continued for 10 minutes to give a dark red solution. Iodomethane (900 μl, 14.40 mmol) was added dropwise and after 1 hour a yellow solid precipitated and about 80% was converted. The mixture was diluted with MeOH and purified by preparative HPLC to give the expected product dimethyl oxazolo [4,5-b] pyridin-2-ylcarbonimidodithioate as a white solid (35 mg, 0 .146 mmol, 2.4% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 2.71 (s, 6 H) 7.22 (dd, J = 8.03, 5.02 Hz, 1 H) 7.72 (dd, J = 8.03, 1.25 Hz, 1 H) 8.49 (dd , J = 5.02, 1.51 Hz, 1 H). MS (LC / MS) RT = 1.358; [M + H] = 240.04.

１０mlのバイアルに、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(８.５３mg, ０.０４４mmol)、ＤＭＦ(２ml)、ＤＩＥＡ(７.７４μｌ, ０.０４mmol)およびＣｓ_２ＣＯ_３(２８.９mg, ０.０８９mmol)を室温で入れ、次にオキサゾロ[４,５−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(１０.６mg, ０.０４４mmol)を入れた。得られた懸濁液を室温で１時間撹拌した。ＬＣ／ＭＳにより、出発物質が完全に消費されたことが示された。反応混合物をＭｅＯＨで希釈し、分取ＨＰＬＣによって精製し、予測された生成物である(Ｒ)−Ｎ−(オキサゾロ[４,５−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の固体として得た(１３mg, ０.０３８mmol, 収率８６％)。
¹H NMR (400 MHz, アセトン-d₆) δ ppm 1.54 - 1.60 (m, 1 H) 1.71 - 1.77 (m, 2 H) 2.16 - 2.24 (m, 1 H) 2.77 - 2.82 (m, 2 H) 2.89 (t, J=7.91 Hz, 4 H) 3.13 - 3.25 (m, 2 H) 3.90 (d, J=10.29 Hz, 1 H) 4.22 (d, J=10.29 Hz, 1 H) 7.13 (dd, J=8.03, 5.02 Hz, 1 H) 7.70 (dd, J=7.91, 1.13 Hz, 1 H) 8.27 (dd, J=5.14, 1.13 Hz, 1 H) 9.11 (br. s., 1 H). MS (LC/MS) R.T. = 0.443; [M+H]⁺ = 300.16。 Step C: (R) -N- (Oxazolo [4,5-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- 2-Amine

To a 10 ml vial was added (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (8.53 mg, 0.044 mmol), DMF (2 ml), DIEA (7.74 μl, 0.04 mmol) and Cs. ₂ CO ₃ (28.9 mg, 0.089 mmol) was charged 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 hour. LC / MS showed that the starting material was completely consumed. The reaction mixture was diluted with MeOH and purified by preparative HPLC and the expected product (R) -N- (oxazolo [4,5-b] pyridin-2-yl) -4H-1′-azaspiro. [Oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white solid (13 mg, 0.038 mmol, 86% yield).
¹ H NMR (400 MHz, acetone-d ₆ ) δ ppm 1.54-1.60 (m, 1 H) 1.71-1.77 (m, 2 H) 2.16-2.24 (m, 1 H) 2.77-2.82 (m, 2 H) 2.89 (t, J = 7.91 Hz, 4 H) 3.13-3.25 (m, 2 H) 3.90 (d, J = 10.29 Hz, 1 H) 4.22 (d, J = 10.29 Hz, 1 H) 7.13 (dd, J = 8.03, 5.02 Hz, 1 H) 7.70 (dd, J = 7.91, 1.13 Hz, 1 H) 8.27 (dd, J = 5.14, 1.13 Hz, 1 H) 9.11 (br. S., 1 H). MS ( LC / MS) RT = 0.443; [M + H] ⁺ = 300.16.

Example 291
(2R) -N- (6,8-Dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole] -2 '-Amine

メタノール(８ml)中の２,２−ジエトキシアセトイミド酸メチル(１.１ｇ, ６.８２mmol)の溶液に、ｐ−トリルメタンアミン(０.７８８ｇ, ６.５０mmol)を環境温度で滴下した。反応フラスコを、予め加熱した油浴に入れ、７０℃で１６時間撹拌し、取り外して冷却した。揮発性成分を減圧下で除去し、粗製の物質を硫酸(５ml)に環境温度で滴下した。反応混合物を７２時間撹拌し、フラスコを氷−水浴に入れ、水(５０ml)で希釈し、水酸化ナトリウム(１０Ｎ)でｐＨ＝１０までゆっくりと中和した。反応混合物が塩基性になったとき、灰色の沈殿物が形成した。この沈殿物を濾過し、水で洗浄し、乾燥させ、６−メチルイソキノリン−３−アミンを灰色の粉末として得た(０.６５ｇ, ６３％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.73 (s, 1 H), 7.69 (d, J=8.28 Hz, 1 H), 7.29 (s, 1 H), 7.00 (d, J=8.28 Hz, 1 H), 5.81 (s, 1 H), 2.40 (s, 3 H). MS (LC/MS) R.T. = 1.37; [M+H]⁺ = 159.10。 Step A: 6-Methylisoquinolin-3-amine

To a solution of methyl 2,2-diethoxyacetimidate (1.1 g, 6.82 mmol) in methanol (8 ml), p-tolylmethanamine (0.788 g, 6.50 mmol) was added dropwise at ambient temperature. The reaction flask was placed in a preheated oil bath, stirred at 70 ° C. for 16 hours, removed and cooled. Volatile components 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 hours, the flask was placed in an ice-water bath, diluted with water (50 ml) and slowly neutralized to pH = 10 with sodium hydroxide (10N). A gray precipitate formed when the reaction mixture became basic. The precipitate was filtered, washed with water and dried to give 6-methylisoquinolin-3-amine as a gray powder (0.65 g, 63%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.73 (s, 1 H), 7.69 (d, J = 8.28 Hz, 1 H), 7.29 (s, 1 H), 7.00 (d, J = 8.28 Hz, 1 H), 5.81 (s, 1 H), 2.40 (s, 3 H). MS (LC / MS) RT = 1.37; [M + H] ⁺ = 159.10.

ジクロロメタン(１０ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(２２０mg, ０.９４８mmol)の溶液に、環境温度で、６−メチルイソキノリン−３−アミン(１２５mg, ０.７９０mmol)を加えた。反応混合物を予め加熱した油浴中に置き、４０℃で１８時間撹拌し、油浴から取り出し、環境温度まで冷却した。混合物を濃縮し、粗製の物質をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中５〜３０％ 酢酸エチル)、３−イソチオシアナト−６−メチルイソキノリンを灰白色の固体として得た(７５mg, ０.３７５mmol, 収率４７.４％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.03 (s, 1 H), 7.88 (d, J=8.24 Hz, 1 H), 7.56 (s, 1 H), 7.46 (dd, J=8.24, 1.53 Hz, 1 H) , 7.39 (s, 1 H) 2.57 (s, 3 H). MS (LC/MS) R.T. = 1.92; [M+H]⁺ = 201.13。 Step B: 3-isothiocyanato-6-methylisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (220 mg, 0.948 mmol) in dichloromethane (10 ml) at ambient temperature 6-methylisoquinolin-3-amine (125 mg, 0.0. 790 mmol) was added. The reaction mixture was placed in a preheated oil bath and stirred at 40 ° C. for 18 hours, removed from the oil bath and cooled to ambient temperature. The mixture was concentrated and the crude material was purified by chromatography on silica gel (5-30% ethyl acetate in hexanes) to give 3-isothiocyanato-6-methylisoquinoline as an off-white solid (75 mg, 0.375 mmol, yield). (Rate 47.4%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.03 (s, 1 H), 7.88 (d, J = 8.24 Hz, 1 H), 7.56 (s, 1 H), 7.46 (dd, J = 8.24, 1.53 Hz, 1 H), 7.39 (s, 1 H) 2.57 (s, 3 H). MS (LC / MS) RT = 1.92; [M + H] ⁺ = 201.13.

Ｎ,Ｎ−ジメチルホルムアミド(６ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(６５.８mg, ０.２８５mmol)に、トリエチルアミン(０.０９０ml, ０.６３mmol)および３−イソチオシアナト−６−メチルイソキノリン(５７mg, ０.２８５mmol)を加えた。懸濁液を、予め加熱した油浴中に置き、７０℃で２時間３０分撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１７７ml, １.１４mmol)を加え、混合物を８５℃で１６時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(クロロホルム中０〜４０％ [９：１ メタノール：水酸化アンモニウム])、次に逆相分取ＨＰＬＣによって精製した(０〜４０％ ＴＦＡ−メタノール−水)。生成物の溶液を、UCT Clean-up CHQAX15M25カートリッジでＭｅＯＨ(３×１０ml)を用いて濾過し、濃縮し、予測された生成物である(Ｓ)−１−((３−ヒドロキシキヌクリジン−３−イル)メチル)−３−(６−メチルイソキノリン−３−イル)チオウレアを黄褐色のゴム状物質として得た。
¹H NMR (400 MHz, MeOD) δ ppm 9.11 (s, 1 H), 8.02 (d, J=8.53 Hz, 1 H), 7.80 (s, 1 H), 7.74 (s, 1 H), 7.47 (d, J=8.53 Hz, 1 H), 4.47 (d, J=10.54 Hz, 1 H), 4.31 (d, J=10.54 Hz, 1 H), 4.10 (d, J=14.81 Hz, 1 H), 3.92 (d, J=14.81 Hz, 1 H), 3.53 - 3.71 (m, 2 H), 3.27 - 3.51 (m, 3 H), 2.84 (br.s, 1 H), 2.55 (s, 3H), 2.48 (m, 1 H), ) 2.10 - 2.30 (m, 3H). MS (LC/MS) R.T. = 1.24; [M+H]⁺ = 323.2。 Step C: (R) -N- (6-Methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (65.8 mg, 0.285 mmol) in N, N-dimethylformamide (6 ml) was added to triethylamine (0.090 ml, 0.63 mmol). And 3-isothiocyanato-6-methylisoquinoline (57 mg, 0.285 mmol) was added. The suspension was placed in a preheated oil bath and stirred at 70 ° C. for 2 hours 30 minutes. N, N′-Diisopropylcarbodiimide (0.177 ml, 1.14 mmol) was added and the mixture was stirred at 85 ° C. for 16 hours. The mixture was concentrated and purified by chromatography on silica gel (0-40% in chloroform [9: 1 methanol: ammonium hydroxide]) and then purified by reverse phase preparative HPLC (0-40% TFA-methanol- water). The product solution was filtered through a UCT Clean-up CHQAX15M25 cartridge using MeOH (3 × 10 ml), concentrated and the expected product (S) -1-((3-hydroxyquinuclidine- 3-yl) methyl) -3- (6-methylisoquinolin-3-yl) thiourea was obtained as a tan gum.
¹ H NMR (400 MHz, MeOD) δ ppm 9.11 (s, 1 H), 8.02 (d, J = 8.53 Hz, 1 H), 7.80 (s, 1 H), 7.74 (s, 1 H), 7.47 ( d, J = 8.53 Hz, 1 H), 4.47 (d, J = 10.54 Hz, 1 H), 4.31 (d, J = 10.54 Hz, 1 H), 4.10 (d, J = 14.81 Hz, 1 H), 3.92 (d, J = 14.81 Hz, 1 H), 3.53-3.71 (m, 2 H), 3.27-3.51 (m, 3 H), 2.84 (br.s, 1 H), 2.55 (s, 3H), 2.48 (m, 1 H),) 2.10-2.30 (m, 3H). MS (LC / MS) RT = 1.24; [M + H] ⁺ = 323.2.

Example 292
(R) -N- (6-Bromoisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(４−ブロモフェニル)メタンアミン塩酸塩(２.３５９ｇ, １０.３９mmol)およびナトリウム メトキシド(２.３７６ml, １０.３９mmol)を、メタノール(１０ml)中の２,２−ジエトキシアセトイミド酸メチル(３.３５ｇ, ２０.７８mmol)の溶液に加えた。濁った混合物を７０℃で１.５時間加熱し、得られた黄色の混合物を濃縮した。残渣をシリカゲルによって精製し、１００％ 酢酸エチル、黄色の粘性の油状物を得た(２.０４ｇ, ６２％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 7.47 (2 H, d, J=8.56 Hz), 7.23 (2 H, d, J=8.06 Hz), 6.76 (1 H, br. s.), 5.31 (1 H, br. s.), 4.94 (1 H, br. s.), 4.45 (2 H, br. s.), 3.47 - 3.77 (4 H, m), 1.26 (6 H, t, J=7.05 Hz). LCMS:R.T. = 2.12; [M+2]⁺ = 317.2。 Step A: N- (4-Bromobenzyl) -2,2-diethoxyacetimidoamide

(4-Bromophenyl) methanamine hydrochloride (2.359 g, 10.39 mmol) and sodium methoxide (2.376 ml, 10.39 mmol) were added to methyl 2,2-diethoxyacetimidate (3 ml) in methanol (10 ml). .35 g, 20.78 mmol). The cloudy mixture was heated at 70 ° C. for 1.5 hours and the resulting yellow mixture was concentrated. The residue was purified by silica gel to give 100% ethyl acetate, a yellow viscous oil (2.04 g, 62%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.47 (2 H, d, J = 8.56 Hz), 7.23 (2 H, d, J = 8.06 Hz), 6.76 (1 H, br.s.), 5.31 (1 H, br. S.), 4.94 (1 H, br. S.), 4.45 (2 H, br. S.), 3.47-3.77 (4 H, m), 1.26 (6 H, t, J = 7.05 Hz). LCMS: RT = 2.12; [M + 2] ⁺ = 317.2.

硫酸(４ml, ９５〜９８％)中のＮ−(４−ブロモベンジル)−２,２−ジエトキシアセトイミドアミド(１.５３ｇ, ４.８５mmol)を、４０℃で１４時間加熱した。混合物を１Ｍ ＮａＯＨでｐＨ ７まで中和し、得られた懸濁液を濾過した。残渣を、シリカゲルのクロマトグラフィーによって、ヘキサン中２０〜５５％ 酢酸エチルで精製した。望ましいフラクションを濃縮し、帯黄褐色の固体を得た(０.４３４ｇ, ４０％)。
LCMS:R.T. = 1.62; [M+2]⁺ = 225.1。 Step B: 6-Bromoisoquinolin-3-amine

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 hours. The mixture was neutralized with 1M NaOH to pH 7 and the resulting suspension was filtered. The residue was purified by chromatography on silica gel with 20-55% ethyl acetate in hexane. The desired fractions were concentrated to give a tan solid (0.434 g, 40%).
LCMS: RT = 1.62; [M + 2] ⁺ = 225.1.

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.２５１ｇ, １.０８０mmol)の溶液に、室温で、６−ブロモイソキノリン−３−アミン(０.２４１ｇ, １.０８０mmol)を加えた。溶液を室温で３時間撹拌した。ＬＣ／ＭＳにより望ましい生成物の形成が示された。深橙色の溶液をシリカゲルのクロマトグラフィーによって精製し(０〜１０％ 酢酸エチル−ヘキサン)、６−ブロモ−３−イソチオシアナトイソキノリンを黄色の油状物として得た(０.１ｇ, ０.３７７mmol, 収率３５％)。
R.T. = 2.54; [M+H]⁺ = 267.04。 Step C: 6-Bromo-3-isothiocyanatoisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.251 g, 1.080 mmol) in dichloromethane at room temperature, 6-bromoisoquinolin-3-amine (0.241 g, 1. 080 mmol) was added. The solution was stirred at room temperature for 3 hours. LC / MS showed the formation of the desired product. The deep orange solution was purified by chromatography on silica gel (0-10% ethyl acetate-hexane) to give 6-bromo-3-isothiocyanatoisoquinoline as a yellow oil (0.1 g, 0.377 mmol, Yield 35%).
RT = 2.54; [M + H] ⁺ = 267.04.

Ｎ,Ｎ−ジメチルホルムアミド(１５ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.０８６ｇ, ０.３７７mmol)に、Ｃｓ_２ＣＯ_３(０.３０７ｇ, ０.９４３mmol)および６−ブロモ−３−イソチオシアナトイソキノリン(０.１ｇ, ０.３７７mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.１７６ml, １.１３２mmol)を加え、混合物をさらに１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(５〜２５％ [９.５：０.５ メタノール：水酸化アンモニウム]−酢酸エチル)、(Ｒ)−Ｎ−(６−ブロモイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.０５４ｇ, ０.１３５mmol, 収率３６％)。
¹H NMR (400 MHz, MeOD) δ ppm 9.04 (1 H, s), 7.80 - 8.05 (2 H, m), 7.55 (1 H, dd, J=8.81, 1.76 Hz), 7.37 (1 H, br. s.), 4.10 (1 H, d, J=10.58 Hz), 3.87 (1 H, d, J=10.83 Hz), 3.68 - 3.77 (1 H, m), 3.56 - 3.67 (1 H, m), 3.29 - 3.49 (4 H, m), 2.45 (1 H, br. s.), 2.28 - 2.41 (1 H, m), 1.86 - 2.15 (3 H, m). LCMS:R.T. = 1.76; [M+]⁺ = 387.21。 Step D: (R) -N- (6-Bromoisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.086 g, 0.377 mmol) in N, N-dimethylformamide (15 ml) was added to Cs ₂ CO ₃ (0.307 g, 0 0.943 mmol) and 6-bromo-3-isothiocyanatoisoquinoline (0.1 g, 0.377 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-diisopropylcarbodiimide (0.176 ml, 1.132 mmol) was added and the mixture was stirred for a further 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-25% [9.5: 0.5 methanol: ammonium hydroxide] -ethyl acetate), (R) -N- (6-bromoisoquinoline-3- Yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.054 g, 0.135 mmol, yield 36). %).
¹ H NMR (400 MHz, MeOD) δ ppm 9.04 (1 H, s), 7.80-8.05 (2 H, m), 7.55 (1 H, dd, J = 8.81, 1.76 Hz), 7.37 (1 H, br s.), 4.10 (1 H, d, J = 10.58 Hz), 3.87 (1 H, d, J = 10.83 Hz), 3.68-3.77 (1 H, m), 3.56-3.67 (1 H, m) , 3.29-3.49 (4 H, m), 2.45 (1 H, br. S.), 2.28-2.41 (1 H, m), 1.86-2.15 (3 H, m). LCMS: RT = 1.76; (M + ] ⁺ = 387.21.

Example 293
(R) -N- (7-bromoisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(３−ブロモフェニル)メタンアミン塩酸塩(３.６２ｇ, １５.６４mmol)およびナトリウム メトキシド(３.５８ml, １５.６４mmol)を、メタノール(１５ml)中の２,２−ジエトキシアセトイミド酸メチル(５.０４２ｇ, ３１.３mmol)の溶液に加えた。濁った混合物を７０℃で１.５時間加熱し、得られた黄色の混合物を濃縮した。残渣を、シリカゲルのクロマトグラフィーによって、１００％ 酢酸エチルで精製した。望ましいフラクションを濃縮し、黄色の粘性の油状物を得た(２.５ｇ, ５１％)。
LCMS: R.T. = 2.11; [M+2]⁺ = 317.06。 Step A: N- (3-bromobenzyl) -2,2-diethoxyacetimidoamide

(3-Bromophenyl) methanamine hydrochloride (3.62 g, 15.64 mmol) and sodium methoxide (3.58 ml, 15.64 mmol) were added to methyl 2,2-diethoxyacetimidate (5 ml in methanol (15 ml)). 0.042 g, 31.3 mmol). The cloudy mixture was heated at 70 ° C. for 1.5 hours and the resulting yellow mixture was concentrated. The residue was purified by chromatography on silica gel with 100% ethyl acetate. The desired fraction was concentrated to give a yellow viscous oil (2.5 g, 51%).
LCMS: RT = 2.11; [M + 2] ⁺ = 317.06.

硫酸(５ml, ９５〜９８％)中のＮ−(３−ブロモベンジル)−２,２−ジエトキシアセトイミドアミド(２.５ｇ, ７.９３mmol)を、４０℃で５４時間加熱した。混合物を１０Ｍ 水性ＮａＯＨで、ｐＨ ７まで中和し、得られた懸濁液を濾過した。残渣を、シリカゲルのクロマトグラフィーによって、ヘキサン中２０〜５５％ 酢酸エチルで、次に１００％ 酢酸エチルで精製した。フラクションを濃縮し、生成物の混合物を含む褐色の固体を得た(１.０ｇ, ５７％)。
LCMS:R.T. = 1.56; [M+2]⁺ = 225.1。
混合物をそのまま次の工程に用いた。 Step B: 7-bromoisoquinolin-3-amine and 5-bromoisoquinolin-3-amine

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 hours. The mixture was neutralized with 10M aqueous NaOH to pH 7, and the resulting suspension was filtered. The residue was purified by chromatography on silica gel with 20-55% ethyl acetate in hexane and then with 100% ethyl acetate. Fractions were concentrated to give a brown solid containing the product mixture (1.0 g, 57%).
LCMS: RT = 1.56; [M + 2] ⁺ = 225.1.
The mixture was used as such for the next step.

ジクロロメタン中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１.１４５ｇ, ４.９３mmol)の溶液に、室温で、７−ブロモイソキノリン−３−アミンおよび５−ブロモイソキノリン−３−アミンの混合物(工程Ｂより)(１.０ｇ, ４.５mmol)を加えた。橙色の溶液を室温で１８時間撹拌した。ＬＣＭＳにより、生成物の形成が示された。深橙色の溶液をシリカゲルのクロマトグラフィーによって精製した(０〜５％ 酢酸エチル−ヘキサン)。最初の生成物のフラクションを合わせて、真空で濃縮し、７−ブロモ−３−イソチオシアナトイソキノリンを黄色の固体として得た(０.２７ｇ, ０.３７７mmol, 収率２２％)。
¹H NMR (400 MHz, アセトン) δ ppm 9.19 (1 H, s), 8.41 (1 H, s), 7.86 - 8.02 (2 H, m), 7.76 (1 H, s). R.T. = 4.28; [M+H]⁺ = 267.04。 Step C: 7-Bromo-3-isothiocyanatoisoquinoline and 5-bromo-3-isothiocyanatoisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (1.145 g, 4.93 mmol) in dichloromethane at room temperature, 7-bromoisoquinolin-3-amine and 5-bromoisoquinoline-3 -A mixture of amines (from Step B) (1.0 g, 4.5 mmol) was added. The orange solution was stirred at room temperature for 18 hours. LCMS indicated product formation. The deep orange solution was purified by silica gel chromatography (0-5% ethyl acetate-hexane). The first product fractions were combined and concentrated in vacuo to give 7-bromo-3-isothiocyanatoisoquinoline as a yellow solid (0.27 g, 0.377 mmol, 22% yield).
¹ H NMR (400 MHz, acetone) δ ppm 9.19 (1 H, s), 8.41 (1 H, s), 7.86-8.02 (2 H, m), 7.76 (1 H, s). RT = 4.28; [ M + H] ⁺ = 267.04.

The second product fractions were combined and concentrated in vacuo to give 5-bromo-3-isothiocyanatoisoquinoline as a yellow solid (0.25 g, 0.377 mmol, 21% yield).
¹ H NMR (400 MHz, acetone) δ ppm 9.24 (1 H, s), 8.23 (1 H, d), 8.15 (1 H, d), 7.81 (1 H, s), 7.64 (1 H, t) LCMS: RT = 4.61; [M + H] ⁺ = 267.04.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２０７ｇ, ０.９０５mmol)に、Ｃｓ_２ＣＯ_３(０.７３７ｇ, ２.２６３mmol)および７−ブロモ−３−イソチオシアナトイソキノリン(０.２４ｇ, ０.９０５mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.４２３ml, ２.７２mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって、酢酸エチル中５〜１５％ [９：１ メタノール：水酸化アンモニウム]を用いて精製した。望ましいフラクションを濃縮し、さらに酢酸エチル中５〜１５％ [９.５：０.５ メタノール：水酸化アンモニウム]を用いて精製し、((Ｒ)−Ｎ−(７−ブロモイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.０６１ｇ, ０.１５６mmol, 収率１７％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.96 (1 H, s), 8.12 (1 H, s), 7.66 (2 H, s), 7.32 (1 H, s), 4.01 (1 H, d), 3.73 (1 H, d), 3.35 - 3.42 (1 H, m), 3.22 - 3.29 (1 H, m), 2.84 - 3.17 (4 H, m), 2.13 - 2.35 (2 H, m), 1.62 - 1.96 (3 H, m). LCMS:R.T. = 1.76; [M+]⁺ = 387.21。 Step D: (R) -N- (7-Bromoisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.207 g, 0.905 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.737 g, 2.263 mmol) and 7-bromo-3-isothiocyanatoisoquinoline (0.24 g, 0.905 mmol) were added. The suspension was stirred at room temperature for 30 minutes. N, N′-Diisopropylcarbodiimide (0.423 ml, 2.72 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel using 5-15% [9: 1 methanol: ammonium hydroxide] in ethyl acetate. The desired fraction was concentrated and further purified using 5-15% in ethyl acetate [9.5: 0.5 methanol: ammonium hydroxide] to give ((R) -N- (7-bromoisoquinolin-3-yl). ) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.061 g, 0.156 mmol, 17% yield). ).
¹ H NMR (400 MHz, MeOD) δ ppm 8.96 (1 H, s), 8.12 (1 H, s), 7.66 (2 H, s), 7.32 (1 H, s), 4.01 (1 H, d) , 3.73 (1 H, d), 3.35-3.42 (1 H, m), 3.22-3.29 (1 H, m), 2.84-3.17 (4 H, m), 2.13-2.35 (2 H, m), 1.62 -1.96 (3 H, m). LCMS: RT = 1.76; [M +] ⁺ = 387.21.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.２０７ｇ, ０.９０５mmol)に、Ｃｓ_２ＣＯ_３(０.７３７ｇ, ２.２６３mmol)および５−ブロモ−３−イソチオシアナトイソキノリン(０.２４ｇ, ０.９０５mmol)を加えた。懸濁液を室温で３０分間撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.４２３ml, ２.７２mmol)を加え、混合物を室温で１８時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製した(５〜１５％ [９.５：０.５ メタノール：水酸化アンモニウム]−酢酸エチル)。望ましいフラクションを濃縮し、さらに酢酸エチル中５〜１５％ [９.５：０.５ メタノール：水酸化アンモニウム]を用いて精製し、(Ｒ)−Ｎ−(５−ブロモイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄色の固体として得た(０.２４８ｇ, ０.６３４mmol, 収率７０％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.99 (1 H, s), 7.91 (2 H, dd), 7.55 (1 H, br. s.), 7.28 (1 H, t), 3.96 (1 H, d), 3.65 (1 H, d), 3.17 - 3.26 (1 H, m), 3.03 - 3.13 (1 H, m), 2.70 - 2.99 (4 H, m), 2.03 - 2.30 (2 H, m), 1.47 - 1.87 (3 H, m). LCMS:R.T. = 1.69; [M+2]⁺ = 389.21。 Example 294
((R) -N- (5-Bromoisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (0.207 g, 0.905 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO 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 added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated and purified by chromatography on silica gel (5-15% [9.5: 0.5 methanol: ammonium hydroxide] -ethyl acetate). The desired fraction was concentrated and further purified using 5-15% in ethyl acetate [9.5: 0.5 methanol: ammonium hydroxide] to give (R) -N- (5-bromoisoquinolin-3-yl). -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained as a yellow solid (0.248 g, 0.634 mmol, 70% yield). .
¹ H NMR (400 MHz, MeOD) δ ppm 8.99 (1 H, s), 7.91 (2 H, dd), 7.55 (1 H, br.s.), 7.28 (1 H, t), 3.96 (1 H , d), 3.65 (1 H, d), 3.17-3.26 (1 H, m), 3.03-3.13 (1 H, m), 2.70-2.99 (4 H, m), 2.03-2.30 (2 H, m ), 1.47-1.87 (3 H, m). LCMS: RT = 1.69; [M + 2] ⁺ = 389.21.

Example 295
(2R) -N- (6,8-Dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole] -2 '-Amine

メタノール(４.９ml)中の２,２−ジエトキシアセトイミド酸メチル(１.５ｇ, ９.３mmol)の溶液に、(２,４−ジメチルフェニル)メタンアミン(１.２ｇ, ８.９mmol)を環境温度で滴下した。反応フラスコを予め加熱した油浴に入れ、７０℃で１６時間撹拌し、冷却し、揮発性成分を減圧下で除去した。粗製の物質を硫酸(１９.７ml)に環境温度で滴下し、７２時間撹拌した。フラスコを氷−水浴に入れ、水(５０ml)で希釈し、水酸化ナトリウム(１０Ｎ)でｐＨ＝１０までゆっくりと中和した。反応混合物を塩基性にしたとき、橙色の沈殿物が形成した。この沈殿物を濾過し、水で洗浄し、乾燥させ、６,８−ジメチルイソキノリン−３−アミンを得た(１.３７ｇ, ７.９５mmol, ９０％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.82 (s, 1 H), 7.17 (s, 1 H), 6.90 (s, 1 H), 6.72 (s, 1 H), 2.61 (s, 3 H), 2.37 (s, 3 H). MS (LC/MS) R.T. = 0.77; [M+H]⁺ = 173.15。 Step A: 6,8-dimethylisoquinolin-3-amine

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). Dropped at ambient temperature. The reaction flask was placed in a preheated oil bath, stirred at 70 ° C. for 16 hours, cooled, and volatile components were removed under reduced pressure. The crude material was added dropwise to sulfuric acid (19.7 ml) at ambient temperature and stirred for 72 hours. The flask was placed in an ice-water bath, diluted with water (50 ml) and slowly neutralized to pH = 10 with sodium hydroxide (10N). An orange precipitate formed when the reaction mixture was basified. The precipitate was filtered, washed with water and dried to give 6,8-dimethylisoquinolin-3-amine (1.37 g, 7.95 mmol, 90%).
¹ H NMR (400 MHz, MeOD) δ ppm 8.82 (s, 1 H), 7.17 (s, 1 H), 6.90 (s, 1 H), 6.72 (s, 1 H), 2.61 (s, 3 H) 2.37 (s, 3 H). MS (LC / MS) RT = 0.77; [M + H] ⁺ = 173.15.

ジクロロメタン(１９ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１.３５ｇ, ５.８mmol)の溶液に、環境温度で、６,８−ジメチルイソキノリン−３−アミン(１ｇ, ５.８mmol)を加えた。反応混合物を予め加熱した油浴中に置き、４０℃で１８時間撹拌し、冷却し、濃縮し、粗製の物質をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中１０〜３５％ 酢酸エチル)、３−イソチオシアナト−６,８−ジメチルイソキノリンを黄色の固体として得た(９３.７mg, ０.４３７mmol, ８％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 9.20 (s, 1 H), 7.36 - 7.44 (m, 2 H), 7.23 (s, 1 H), 2.75 (s, 3 H), 2.51 (s, 3 H). MS (LC/MS) R.T. = 2.03; [M+H]⁺ = 215.1。 Step B: 3-isothiocyanato-6,8-dimethylisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (1.35 g, 5.8 mmol) in dichloromethane (19 ml) at ambient temperature, 6,8-dimethylisoquinolin-3-amine ( 1 g, 5.8 mmol) was added. The reaction mixture was placed in a preheated oil bath, stirred at 40 ° C. for 18 hours, cooled and concentrated, and the crude material was purified by chromatography on silica gel (10-35% ethyl acetate in hexane), 3- Isothiocyanato-6,8-dimethylisoquinoline was obtained as a yellow solid (93.7 mg, 0.437 mmol, 8%).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 9.20 (s, 1 H), 7.36-7.44 (m, 2 H), 7.23 (s, 1 H), 2.75 (s, 3 H), 2.51 (s, 3 H). MS (LC / MS) RT = 2.03; [M + H] ⁺ = 215.1.

Ｎ,Ｎ−ジメチルホルムアミド(１.４ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(９９mg, ０.４３mmol)に、トリエチルアミン(０.１８ml, １.３mmol)および３−イソチオシアナト−６,８−ジメチルイソキノリン(９３mg, ０.４３mmol)を加えた。懸濁液を、予め加熱した油浴中に置き、７０℃で２時間３０分撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.２７ml, １.７mmol)を加え、混合物を８５℃で１６時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(クロロホルム中０〜４０％ [９：１ メタノール：水酸化アンモニウム])、次に逆相分取ＨＰＬＣによって精製し(０〜４０％ [０.１％ ＴＦＡ]−メタノール−水)、トリフルオロ酢酸塩として(２Ｒ)−Ｎ−(６,８−ジメチル−３−イソキノリニル)−４'Ｈ−スピロ[４−アザビシクロ[２.２.２]オクタン−２,５'−[１,３]オキサゾール]−２'−アミンを白色の固体として得た(２４mg, ０.０５３mmol, 収率１２％)。
¹H NMR (400 MHz, MeOD) δ ppm 9.29 (s, 1 H), 7.54 (s, 1 H), 7.43 (s, 1 H), 7.34 (s, 1 H), 4.38 (d, J=11.04 Hz, 1 H), 4.17 (d, J=11.04 Hz, 1 H), 3.94- 4.10 (m, 1 H), 3.86 (dd, J=15.06, 2.26 Hz, 1 H), 3.51 - 3.67 (m, 1 H), 3.37 - 3.51 (m, 3 H), 2.77 (s, 3 H), 2.73 (d, J=3.51 Hz, 1 H), 2.48 - 2.57 (m, 3 H), 2.30 - 2.46 (m, 1 H), 1.94 - 2.28 (m, 3 H). MS (LC/MS) R.T. = 0.90; [M+H]⁺ = 337.38。 Step C: (2R) -N- (6,8-Dimethyl-3-isoquinolinyl) -4′H-spiro [4-azabicyclo [2.2.2] octane-2,5 ′-[1,3] oxazole ] -2'-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (99 mg, 0.43 mmol) in N, N-dimethylformamide (1.4 ml) was added to triethylamine (0.18 ml, 1.3 mmol). ) And 3-isothiocyanato-6,8-dimethylisoquinoline (93 mg, 0.43 mmol). The suspension was placed in a preheated oil bath and stirred at 70 ° C. for 2 hours 30 minutes. N, N′-diisopropylcarbodiimide (0.27 ml, 1.7 mmol) was added and the mixture was stirred at 85 ° C. for 16 hours. The mixture was concentrated and purified by chromatography on silica gel (0-40% in chloroform [9: 1 methanol: ammonium hydroxide]), then purified by reverse phase preparative HPLC (0-40% [0.1 % TFA] -methanol-water), (2R) -N- (6,8-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane- as trifluoroacetate 2,5 ′-[1,3] oxazole] -2′-amine was obtained as a white solid (24 mg, 0.053 mmol, 12% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 9.29 (s, 1 H), 7.54 (s, 1 H), 7.43 (s, 1 H), 7.34 (s, 1 H), 4.38 (d, J = 11.04 Hz, 1 H), 4.17 (d, J = 11.04 Hz, 1 H), 3.94-4.10 (m, 1 H), 3.86 (dd, J = 15.06, 2.26 Hz, 1 H), 3.51-3.67 (m, 1 H), 3.37-3.51 (m, 3 H), 2.77 (s, 3 H), 2.73 (d, J = 3.51 Hz, 1 H), 2.48-2.57 (m, 3 H), 2.30-2.46 (m , 1 H), 1.94-2.28 (m, 3 H). MS (LC / MS) RT = 0.90; [M + H] ⁺ = 337.38.

(Ｒ)−Ｎ−(３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、ピリジン−３−イルボロン酸および塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.95 (br. s., 2 H), 8.66 (d, J=4.6 Hz, 1 H), 8.31 (d, J=5.5 Hz, 1 H), 8.14 (br. s., 1 H), 7.54 (d, J=7.6 Hz, 1 H), 7.26 (br. s., 2 H), 3.85 (br. s., 1 H), 3.58 (s, 1 H), 3.00 (br. s., 2 H), 2.67 (br. s., 4 H), 1.90 (br. s., 2 H), 1.59 (br. s., 2 H), 1.45 (br. s., 1 H). MS (LC/MS) R.T. = 0.12; [M+H]⁺ = 336.18。 Example 296
(R) -N- (3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine From the 4-bromopyridin-2-amine, pyridin-3-ylboronic acid and 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex, the general procedure of Example 28, steps AB Manufactured according to the procedure.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.95 (br. S., 2 H), 8.66 (d, J = 4.6 Hz, 1 H), 8.31 (d, J = 5.5 Hz, 1 H) , 8.14 (br. S., 1 H), 7.54 (d, J = 7.6 Hz, 1 H), 7.26 (br. S., 2 H), 3.85 (br. S., 1 H), 3.58 (s , 1 H), 3.00 (br. S., 2 H), 2.67 (br. S., 4 H), 1.90 (br. S., 2 H), 1.59 (br. S., 2 H), 1.45 (br. s., 1 H). MS (LC / MS) RT = 0.12; [M + H] ⁺ = 336.18.

(Ｒ)−Ｎ−(５−クロロ−３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、５−クロロピリジン−３−イルボロン酸および塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.92 (s, 2 H), 8.71 (d, J=2.1 Hz, 1 H), 8.19 - 8.39 (m, 2 H), 7.30 (d, J=4.3 Hz, 2 H), 3.77 - 3.95 (m, 1 H), 3.59 (d, J=10.1 Hz, 1 H), 3.01 (d, J=4.9 Hz, 2 H), 2.61 - 2.86 (m, 4 H), 2.00 (s, 1 H), 1.83 - 1.96 (m, 1 H), 1.55 - 1.68 (m, 2 H), 1.40 - 1.54 (m, 1 H). MS (LC/MS) R.T. = 0.91; [M+H]⁺ = 370.09。 Example 297
(R) -N- (5-Chloro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (5-Chloro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 Step of Example 28 from 4-bromopyridin-2-amine, 5-chloropyridin-3-ylboronic acid and 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex Prepared according to the general procedure of AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.92 (s, 2 H), 8.71 (d, J = 2.1 Hz, 1 H), 8.19-8.39 (m, 2 H), 7.30 (d, J = 4.3 Hz, 2 H), 3.77-3.95 (m, 1 H), 3.59 (d, J = 10.1 Hz, 1 H), 3.01 (d, J = 4.9 Hz, 2 H), 2.61-2.86 (m, 4 H), 2.00 (s, 1 H), 1.83-1.96 (m, 1 H), 1.55-1.68 (m, 2 H), 1.40-1.54 (m, 1 H). MS (LC / MS) RT = 0.91; [M + H] ⁺ = 370.09.

(Ｒ)−Ｎ−(６−メトキシ−３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、６−メトキシピリジン−３−イルボロン酸および塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.79 - 9.21 (m, 1 H), 8.46 - 8.66 (m, 1 H), 8.26 (s, 1 H), 7.97 - 8.14 (m, 1 H), 6.99 - 7.27 (m, 2 H), 6.83 - 6.99 (m, 1 H), 3.92 (s, 4 H), 3.49 - 3.62 (m, 1 H), 2.99 (d, J=4.3 Hz, 2 H), 2.66 (d, J=7.6 Hz, 4 H), 1.79 - 2.05 (m, 2 H), 1.33 - 1.63 (m, 3 H). MS (LC/MS) R.T. = 0.98; [M+H]⁺ = 366.17。 Example 298
(R) -N- (6-Methoxy-3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (6-Methoxy-3,4'-bipyridin-2'-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2 Step of Example 28 from 4-bromopyridin-2-amine, 6-methoxypyridin-3-ylboronic acid and 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex Prepared according to the general procedure of AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.79-9.21 (m, 1 H), 8.46-8.66 (m, 1 H), 8.26 (s, 1 H), 7.97-8.14 (m, 1 H ), 6.99-7.27 (m, 2 H), 6.83-6.99 (m, 1 H), 3.92 (s, 4 H), 3.49-3.62 (m, 1 H), 2.99 (d, J = 4.3 Hz, 2 H), 2.66 (d, J = 7.6 Hz, 4 H), 1.79-2.05 (m, 2 H), 1.33-1.63 (m, 3 H). MS (LC / MS) RT = 0.98; (M + H ] ⁺ = 366.17.

(Ｒ)−Ｎ−(６−フルオロ−３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、２−フルオロ−５−(４,４,５,５−テトラメチル−１,３,２−ジオキサボロラン−２−イル)ピリジンおよび塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.90 - 9.17 (m, 1 H), 8.57 - 8.71 (m, 1 H), 8.31 (d, J=5.5 Hz, 2 H), 7.31 - 7.40 (m, 1 H), 7.01 - 7.29 (m, 2 H), 3.76 - 3.95 (m, 1 H), 3.59 (d, J=10.4 Hz, 1 H), 3.01 (br. s., 2 H), 2.68 (br. s., 4 H), 1.95 - 2.05 (m, 1 H), 1.81 - 1.95 (m, 1 H), 1.59 (br. s., 2 H), 1.37 - 1.53 (m, 1 H). MS (LC/MS) R.T. = 0.68; [M+H]⁺ = 354.09。 Example 299
(R) -N- (6-Fluoro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (6-Fluoro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 The amine is 4-bromopyridin-2-amine, 2-fluoro-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine and 1,1 ′ chloride Prepared from the bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.90-9.17 (m, 1 H), 8.57-8.71 (m, 1 H), 8.31 (d, J = 5.5 Hz, 2 H), 7.31-7.40 (m, 1 H), 7.01-7.29 (m, 2 H), 3.76-3.95 (m, 1 H), 3.59 (d, J = 10.4 Hz, 1 H), 3.01 (br. s., 2 H) , 2.68 (br. S., 4 H), 1.95-2.05 (m, 1 H), 1.81-1.95 (m, 1 H), 1.59 (br. S., 2 H), 1.37-1.53 (m, 1 H). MS (LC / MS) RT = 0.68; [M + H] ⁺ = 354.09.

(Ｒ)−Ｎ−(４−(３−クロロ−４−フルオロ−フェニル)ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、３−クロロ−４−フルオロフェニルボロン酸および塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.92 - 9.16 (m, 1 H), 8.28 (d, J=5.2 Hz, 1 H), 7.88 - 8.09 (m, 1 H), 7.66 - 7.81 (m, 1 H), 7.43 - 7.63 (m, 1 H), 7.19 - 7.30 (m, 1 H), 6.99 - 7.17 (m, 1 H), 3.77 - 3.94 (m, 1 H), 3.53 - 3.66 (m, 1 H), 2.99 (br. s., 2 H), 2.67 (br. s., 4 H), 1.82 - 2.03 (m, 2 H), 1.52 - 1.66 (m, 2 H), 1.37 - 1.53 (m, 1 H). MS (LC/MS) R.T. = 1.65; [M+H]⁺ = 387.10。 Example 300
(R) -N- (4- (3-Chloro-4-fluoro-phenyl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

(R) -N- (4- (3-Chloro-4-fluoro-phenyl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-amine was prepared from 4-bromopyridin-2-amine, 3-chloro-4-fluorophenylboronic acid and 1,1'-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex Prepared according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.92-9.16 (m, 1 H), 8.28 (d, J = 5.2 Hz, 1 H), 7.88-8.09 (m, 1 H), 7.66-7.81 (m, 1 H), 7.43-7.63 (m, 1 H), 7.19-7.30 (m, 1 H), 6.99-7.17 (m, 1 H), 3.77-3.94 (m, 1 H), 3.53-3.66 (m, 1 H), 2.99 (br. s., 2 H), 2.67 (br. s., 4 H), 1.82-2.03 (m, 2 H), 1.52-1.66 (m, 2 H), 1.37 -1.53 (m, 1 H). MS (LC / MS) RT = 1.65; [M + H] ⁺ = 387.10.

(Ｒ)−Ｎ−(４−ｍ−トリルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、ｍ−トリルボロン酸および塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.86 - 9.25 (m, 1 H), 8.26 (d, J=5.5 Hz, 1 H), 7.45 - 7.57 (m, 2 H), 7.39 (t, J=7.5 Hz, 1 H), 7.27 (d, J=7.3 Hz, 1 H), 7.18 (d, J=4.3 Hz, 2 H), 3.76 - 3.98 (m, 1 H), 3.58 (d, J=10.4 Hz, 1 H), 2.99 (d, J=5.8 Hz, 2 H), 2.72 - 2.85 (m, 2 H), 2.67 (t, J=7.6 Hz, 2 H), 2.39 (s, 3 H), 1.83 - 2.05 (m, 2 H), 1.52 - 1.64 (m, 2 H), 1.36 - 1.51 (m, 1 H). MS (LC/MS) R.T. = 1.54; [M+H]⁺ = 349.19。 Example 301
(R) -N- (4-m-Tolylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4-m-Tolylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 4 Prepared from the bromopyridin-2-amine, m-tolylboronic acid and 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex according to the general procedure of Example 28, steps AB did.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.86-9.25 (m, 1 H), 8.26 (d, J = 5.5 Hz, 1 H), 7.45-7.57 (m, 2 H), 7.39 (t , J = 7.5 Hz, 1 H), 7.27 (d, J = 7.3 Hz, 1 H), 7.18 (d, J = 4.3 Hz, 2 H), 3.76-3.98 (m, 1 H), 3.58 (d, J = 10.4 Hz, 1 H), 2.99 (d, J = 5.8 Hz, 2 H), 2.72-2.85 (m, 2 H), 2.67 (t, J = 7.6 Hz, 2 H), 2.39 (s, 3 H), 1.83-2.05 (m, 2 H), 1.52-1.64 (m, 2 H), 1.36-1.51 (m, 1 H). MS (LC / MS) RT = 1.54; [M + H] ⁺ = 349.19.

(Ｒ)−Ｎ−(４−フェニルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミンから、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.91 - 9.29 (m, 1 H), 8.27 (d, J=5.5 Hz, 1 H), 7.73 (d, J=7.3 Hz, 2 H), 7.38 - 7.58 (m, 3 H), 6.92 - 7.27 (m, 2 H), 3.78 - 3.94 (m, 1 H), 3.59 (d, J=10.4 Hz, 1 H), 2.91 - 3.08 (m, 2 H), 2.59 - 2.87 (m, 4 H), 1.82 - 2.05 (m, 2 H), 1.59 (br. s., 2 H), 1.36 - 1.53 (m, 1 H). MS (LC/MS) R.T. = 1.24; [M+H]⁺ = 335.15。 Example 302
(R) -N- (4-Phenylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4-Phenylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is converted to 4-bromo Prepared from pyridin-2-amine according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.91-9.29 (m, 1 H), 8.27 (d, J = 5.5 Hz, 1 H), 7.73 (d, J = 7.3 Hz, 2 H), 7.38-7.58 (m, 3 H), 6.92-7.27 (m, 2 H), 3.78-3.94 (m, 1 H), 3.59 (d, J = 10.4 Hz, 1 H), 2.91-3.08 (m, 2 H), 2.59-2.87 (m, 4 H), 1.82-2.05 (m, 2 H), 1.59 (br. S., 2 H), 1.36-1.53 (m, 1 H). MS (LC / MS) RT = 1.24; [M + H] ⁺ = 335.15.

(Ｒ)−Ｎ−(４−(１−メチル−１Ｈ−ピラゾール−４−イル)ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、１−メチル−４−(４,４,５,５−テトラメチル−１,３,２−ジオキサボロラン−２−イル)−１Ｈ−ピラゾールおよび塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.80 - 9.17 (m, 1 H), 8.26 (s, 1 H), 8.14 (d, J=5.2 Hz, 1 H), 7.94 (br. s., 1 H), 7.06 (d, J=4.3 Hz, 2 H), 3.88 (s, 4 H), 3.56 (d, J=9.8 Hz, 1 H), 2.98 (br. s., 2 H), 2.60 - 2.86 (m, 4 H), 1.79 - 2.07 (m, 2 H), 1.58 (br. s., 2 H), 1.46 (dd, J=9.6, 2.6 Hz, 1 H). MS (LC/MS) R.T. = 0.7; [M+H]⁺ = 339.18。 Example 303
(R) -N- (4- (1-Methyl-1H-pyrazol-4-yl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2 ] Octane] -2-amine

(R) -N- (4- (1-Methyl-1H-pyrazol-4-yl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2 ] Octane] -2-amine with 4-bromopyridin-2-amine, 1-methyl-4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H -Prepared from the pyrazole and 1,1'-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex according to the general procedure of steps 28-B of Example 28.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.80-9.17 (m, 1 H), 8.26 (s, 1 H), 8.14 (d, J = 5.2 Hz, 1 H), 7.94 (br. S ., 1 H), 7.06 (d, J = 4.3 Hz, 2 H), 3.88 (s, 4 H), 3.56 (d, J = 9.8 Hz, 1 H), 2.98 (br. S., 2 H) , 2.60-2.86 (m, 4 H), 1.79-2.07 (m, 2 H), 1.58 (br. S., 2 H), 1.46 (dd, J = 9.6, 2.6 Hz, 1 H). MS (LC / MS) RT = 0.7; [M + H] ⁺ = 339.18.

(Ｒ)−Ｎ−(４−(チアゾール−４−イル)ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、４−(トリブチルスタンニル)チアゾールおよび塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.25 (d, J=1.8 Hz, 2 H), 8.40 (s, 1 H), 8.27 (d, J=5.5 Hz, 1 H), 7.23 - 7.53 (m, 2 H), 3.85 (br. s., 1 H), 3.58 (d, J=10.4 Hz, 1 H), 3.00 (d, J=6.7 Hz, 2 H), 2.73 - 2.85 (m, 2 H), 2.68 (t, J=7.8 Hz, 2 H), 1.84 - 2.04 (m, 2 H), 1.53 - 1.68 (m, 2 H), 1.47 (d, J=7.0 Hz, 1 H). MS (LC/MS) R.T. = 0.78; [M+H]⁺ = 342.17。 Example 304
(R) -N- (4-thiazol-4-yl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4- (thiazol-4-yl) pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- The amine was obtained from 4-bromopyridin-2-amine, 4- (tributylstannyl) thiazole and 1,1′-bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex from Example 28, steps A- Prepared according to the general procedure for B.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.25 (d, J = 1.8 Hz, 2 H), 8.40 (s, 1 H), 8.27 (d, J = 5.5 Hz, 1 H), 7.23- 7.53 (m, 2 H), 3.85 (br. S., 1 H), 3.58 (d, J = 10.4 Hz, 1 H), 3.00 (d, J = 6.7 Hz, 2 H), 2.73-2.85 (m , 2 H), 2.68 (t, J = 7.8 Hz, 2 H), 1.84-2.04 (m, 2 H), 1.53-1.68 (m, 2 H), 1.47 (d, J = 7.0 Hz, 1 H) MS (LC / MS) RT = 0.78; [M + H] ⁺ = 342.17.

(Ｒ)−Ｎ−(６−ニトロ−３,４'−ビピリジン−２'−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、４−ブロモピリジン−２−アミン、２−ニトロ−５−(４,４,５,５−テトラメチル−１,３,２−ジオキサボロラン−２−イル)ピリジンおよび塩化 １,１'−ビス(ジフェニルホスフィノ)フェロセン−パラジウム(II)ジクロロメタン錯体から、実施例２８の工程Ａ〜Ｂの一般的な手順に従って製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.06 (br. s., 2 H), 8.52 - 8.67 (m, 1 H), 8.31 - 8.46 (m, 2 H), 7.13 - 7.44 (m, 2 H), 3.78 - 3.94 (m, 1 H), 3.60 (d, J=10.1 Hz, 1 H), 3.00 (br. s., 2 H), 2.59 - 2.84 (m, 4 H), 2.00 (br. s., 2 H), 1.59 (br. s., 2 H), 1.38 - 1.53 (m, 1 H). MS (LC/MS) R.T. = 0.81; [M+H]⁺ = 381.2。 Example 305
(R) -N- (6-Nitro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (6-Nitro-3,4′-bipyridin-2′-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 The amine is 4-bromopyridin-2-amine, 2-nitro-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine and 1,1 ′ chloride Prepared from the bis (diphenylphosphino) ferrocene-palladium (II) dichloromethane complex according to the general procedure of Example 28, Steps AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.06 (br. S., 2 H), 8.52-8.67 (m, 1 H), 8.31-8.46 (m, 2 H), 7.13-7.44 (m , 2 H), 3.78-3.94 (m, 1 H), 3.60 (d, J = 10.1 Hz, 1 H), 3.00 (br. S., 2 H), 2.59-2.84 (m, 4 H), 2.00 (br. s., 2 H), 1.59 (br. s., 2 H), 1.38-1.53 (m, 1 H). MS (LC / MS) RT = 0.81; [M + H] ⁺ = 381.2.

(Ｒ)−Ｎ−(５−クロロ−６−メチルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、５−クロロ−６−メチルピリジン−２−アミンを出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.81 (br. s., 1 H), 7.59 (br. s., 1 H), 6.64 (br. s., 1 H), 3.82 (br. s., 1 H), 3.56 (d, J=10.4 Hz, 1 H), 2.97 (s, 2 H), 2.58 - 2.82 (m, 4 H), 2.48 (br. s., 3 H), 1.98 (br. s., 1 H), 1.81 - 1.93 (m, 1 H), 1.58 (br. s., 2 H), 1.28 - 1.51 (m, 1 H). MS (LC/MS) R.T. = 0.89; [M+H]⁺ = 307.08。 Example 306
(R) -N- (5-Chloro-6-methylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Chloro-6-methylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine Prepared according to the general procedure of Example 23, Steps AB, using 5-chloro-6-methylpyridin-2-amine as the starting material.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.81 (br. S., 1 H), 7.59 (br. S., 1 H), 6.64 (br. S., 1 H), 3.82 (br s., 1 H), 3.56 (d, J = 10.4 Hz, 1 H), 2.97 (s, 2 H), 2.58-2.82 (m, 4 H), 2.48 (br. s., 3 H), 1.98 (br. S., 1 H), 1.81-1.93 (m, 1 H), 1.58 (br. S., 2 H), 1.28-1.51 (m, 1 H). MS (LC / MS) RT = 0.89; [M + H] ⁺ = 307.08.

(Ｒ)−Ｎ−(５−フルオロ−４−メチルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、５−フルオロ−４−メチルピリジン−２−アミンを出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.59 - 8.97 (m, 1 H), 8.04 (s, 1 H), 6.50 - 6.95 (m, 1 H), 3.79 (br. s., 1 H), 3.53 (d, J=10.4 Hz, 1 H), 2.96 (br. s., 2 H), 2.60 - 2.84 (m, 4 H), 2.21 (s, 3 H), 1.95 (br. s., 2 H), 1.52 - 1.66 (m, 2 H), 1.45 (ddd, J=6.9, 2.9, 2.7 Hz, 1 H). MS (LC/MS) R.T. = 0.63; [M+H]⁺ = 291.12。 Example 307
(R) -N- (5-Fluoro-4-methylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Fluoro-4-methylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared according to the general procedure of Example 23, Steps AB, using 5-fluoro-4-methylpyridin-2-amine as the starting material.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.59-8.97 (m, 1 H), 8.04 (s, 1 H), 6.50-6.95 (m, 1 H), 3.79 (br. S., 1 H), 3.53 (d, J = 10.4 Hz, 1 H), 2.96 (br.s., 2 H), 2.60-2.84 (m, 4 H), 2.21 (s, 3 H), 1.95 (br. S ., 2 H), 1.52-1.66 (m, 2 H), 1.45 (ddd, J = 6.9, 2.9, 2.7 Hz, 1 H). MS (LC / MS) RT = 0.63; [M + H] ⁺ = 291.12.

(Ｒ)−Ｎ−(３,５−ジクロロ−６−メチルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、３,５−ジクロロ−６−メチルピリジン−２−アミンを出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.83 (br. s., 1 H), 7.86 (d, J=2.1 Hz, 1 H), 3.85 (d, J=9.8 Hz, 1 H), 3.59 (d, J=10.1 Hz, 1 H), 3.00 (br. s., 1 H), 2.73 - 2.84 (m, 1 H), 2.67 (t, J=7.5 Hz, 1 H), 2.49 (dd, J=14.5, 2.0 Hz, 6 H), 2.01 (br. s., 1 H), 1.89 (br. s., 1 H), 1.59 (br. s., 2 H), 1.37 - 1.51 (m, 1 H). MS (LC/MS) R.T. = 1.07; [M+H]⁺ = 341.06。 Example 308
(R) -N- (3,5-Dichloro-6-methylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (3,5-Dichloro-6-methylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- The amine was prepared according to the general procedure of Example 23, Steps AB using 3,5-dichloro-6-methylpyridin-2-amine as the starting material.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.83 (br. S., 1 H), 7.86 (d, J = 2.1 Hz, 1 H), 3.85 (d, J = 9.8 Hz, 1 H) , 3.59 (d, J = 10.1 Hz, 1 H), 3.00 (br. S., 1 H), 2.73-2.84 (m, 1 H), 2.67 (t, J = 7.5 Hz, 1 H), 2.49 ( dd, J = 14.5, 2.0 Hz, 6 H), 2.01 (br. s., 1 H), 1.89 (br. s., 1 H), 1.59 (br. s., 2 H), 1.37-1.51 ( m, 1 H). MS (LC / MS) RT = 1.07; [M + H] ⁺ = 341.06.

(Ｒ)−Ｎ−(４,５−ジメチルピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、４,５−ジメチルピリジン−２−アミンを出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.71 - 9.29 (m, 1 H), 7.92 (s, 1 H), 6.28 - 7.02 (m, 1 H), 3.80 (br. s., 1 H), 3.53 (d, J=10.1 Hz, 1 H), 2.87 - 3.03 (m, 2 H), 2.59 - 2.85 (m, 4 H), 2.04 - 2.26 (m, 6 H), 1.80 - 1.99 (m, 2 H), 1.33 - 1.64 (m, 3 H). MS (LC/MS) R.T. = 0.67; [M+H]⁺ = 287.20。 Example 309
(R) -N- (4,5-Dimethylpyridin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4,5-dimethylpyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared according to the general procedure of Example 23, Steps AB, using 4,5-dimethylpyridin-2-amine as the starting material.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.71-9.29 (m, 1 H), 7.92 (s, 1 H), 6.28-7.02 (m, 1 H), 3.80 (br. S., 1 H), 3.53 (d, J = 10.1 Hz, 1 H), 2.87-3.03 (m, 2 H), 2.59-2.85 (m, 4 H), 2.04-2.26 (m, 6 H), 1.80-1.99 ( m, 2 H), 1.33-1.64 (m, 3 H). MS (LC / MS) RT = 0.67; [M + H] ⁺ = 287.20.

(Ｒ)−Ｎ−(５−メトキシピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３の工程Ａ〜Ｂの一般的な手順に従って、５−メトキシピリジン−２−アミンを出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.57 - 9.10 (m, 1 H), 7.91 (d, J=3.1 Hz, 1 H), 7.30 (dd, J=8.9, 3.4 Hz, 1 H), 6.54 - 7.04 (m, 1 H), 3.72 - 3.85 (m, 4 H), 3.52 (d, J=10.1 Hz, 1 H), 2.88 - 3.03 (m, 2 H), 2.76 (d, J=5.8 Hz, 2 H), 2.66 (t, J=7.6 Hz, 2 H), 1.81 - 2.02 (m, 2 H), 1.51 - 1.64 (m, 2 H), 1.38 - 1.49 (m, 1 H). MS (LC/MS) R.T. = 0.46; [M+H]⁺ = 289.17。 Example 310
(R) -N- (5-Methoxypyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Methoxypyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared according to Example 23. Prepared using 5-methoxypyridin-2-amine as the starting material according to the general procedure of Steps AB.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.57-9.10 (m, 1 H), 7.91 (d, J = 3.1 Hz, 1 H), 7.30 (dd, J = 8.9, 3.4 Hz, 1 H ), 6.54-7.04 (m, 1 H), 3.72-3.85 (m, 4 H), 3.52 (d, J = 10.1 Hz, 1 H), 2.88-3.03 (m, 2 H), 2.76 (d, J = 5.8 Hz, 2 H), 2.66 (t, J = 7.6 Hz, 2 H), 1.81-2.02 (m, 2 H), 1.51-1.64 (m, 2 H), 1.38-1.49 (m, 1 H) MS (LC / MS) RT = 0.46; [M + H] ⁺ = 289.17.

Example 311
(R) -N- (4-Chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

メタノール(６０ml)中のイソキノリン−３−アミン(２.０２ｇ, １４.０mmol)に、Ｎ−クロロスクシンイミド(２.１９ｇ, １６.４mmol)を室温で加えた。反応物を室温で３時間撹拌し、真空で濃縮した。粗製の物質についてクロマトグラフィーを行い(Biotage, 10〜１００％ 酢酸エチル／ヘキサン)、４−クロロイソキノリン−３−アミンを固体として得た(２.１ｇ, １１.８mmol, 収率８４％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.86 (s, 1 H), 7.93 (d, J=7.6 Hz, 1 H), 7.79 (d, J=8.5 Hz, 1 H), 7.68 (td, J=6.7, 1.8 Hz, 1 H), 7.24 - 7.35 (m, 1 H), 6.32 (br. s., 2 H). MS (LC/MS) R.T. = 1.50; [M+H]⁺ = 178.96。 Step A: 4-Chloroisoquinolin-3-amine

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 and concentrated in vacuo. The crude material was chromatographed (Biotage, 10-100% ethyl acetate / hexane) to give 4-chloroisoquinolin-3-amine as a solid (2.1 g, 11.8 mmol, 84% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.86 (s, 1 H), 7.93 (d, J = 7.6 Hz, 1 H), 7.79 (d, J = 8.5 Hz, 1 H), 7.68 ( td, J = 6.7, 1.8 Hz, 1 H), 7.24-7.35 (m, 1 H), 6.32 (br. s., 2 H). MS (LC / MS) RT = 1.50; [M + H] ⁺ = 178.96.

(Ｒ)−Ｎ−(４−クロロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の工程Ａ〜Ｂに従って、４−クロロイソキノリン−３−アミン(上記の工程Ａより)を出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.98 - 9.10 (m, 1 H), 8.57 - 8.95 (m, 1 H), 7.99 - 8.18 (m, 2 H), 7.79 (s, 1 H), 7.51 (s, 1 H), 3.84 (d, J=9.8 Hz, 1 H), 3.35 - 3.51 (m, 1 H), 3.02 (d, J=5.5 Hz, 2 H), 2.78 (s, 2 H), 2.61 - 2.73 (m, 2 H), 2.00 (s, 2 H), 1.55 - 1.65 (m, 2 H), 1.42 - 1.54 (m, 1 H). MS (LC/MS) R.T. = 0.92; [M+H]⁺ = 343.09。 Step B: (R) -N- (4-Chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4-Chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 267. Prepared using 4-chloroisoquinolin-3-amine (from Step A above) as a starting material according to Steps A-B.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.98-9.10 (m, 1 H), 8.57-8.95 (m, 1 H), 7.99-8.18 (m, 2 H), 7.79 (s, 1 H ), 7.51 (s, 1 H), 3.84 (d, J = 9.8 Hz, 1 H), 3.35-3.51 (m, 1 H), 3.02 (d, J = 5.5 Hz, 2 H), 2.78 (s, 2 H), 2.61-2.73 (m, 2 H), 2.00 (s, 2 H), 1.55-1.65 (m, 2 H), 1.42-1.54 (m, 1 H). MS (LC / MS) RT = 0.92; [M + H] ⁺ = 343.09.

Example 312
(R) -N- (7-chloro-8-fluoroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(３−クロロ−２−フルオロフェニル)メタンアミン(１.０ｇ, ６.２７mmol)を、メタノール(１０ml)中の２,２−ジエトキシアセトイミド酸メチル(１.１２ｇ, ６.９６mmol)の溶液に加えた。混合物を７０℃で１時間加熱した。混合物をクロマトグラフィーによって精製し(Biotage, １００％ 酢酸エチル)、Ｎ−(３−クロロ−２−フルオロベンジル)−２,２−ジエトキシアセトイミドアミドを無色の粘性の油状物として得た(１.３ｇ, ４.５mmol, 収率６５％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 7.23 - 7.40 (m, 2 H), 7.07 (t, J=7.9 Hz, 1 H), 4.93 (s, 1 H), 4.54 (s, 2 H), 3.50 - 3.74 (m, 4 H), 1.18 - 1.35 (m, 6 H). MS (LC/MS) R.T. = 1.78; [M+H]⁺ = 289.17。 Step A: N- (3-Chloro-2-fluorobenzyl) -2,2-diethoxyacetimidoamide

(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). added. The mixture was heated at 70 ° C. for 1 hour. The mixture was purified by chromatography (Biotage, 100% ethyl acetate) to give N- (3-chloro-2-fluorobenzyl) -2,2-diethoxyacetimidoamide as a colorless viscous oil (1 .3 g, 4.5 mmol, 65% yield).
¹ H NMR (500 MHz, chloroform-d) δ ppm 7.23-7.40 (m, 2 H), 7.07 (t, J = 7.9 Hz, 1 H), 4.93 (s, 1 H), 4.54 (s, 2 H ), 3.50-3.74 (m, 4 H), 1.18-1.35 (m, 6 H). MS (LC / MS) RT = 1.78; [M + H] ⁺ = 289.17.

Ｎ−(３−クロロ−２−フルオロベンジル)−２,２−ジエトキシアセトイミドアミド(０.９７ｇ, ３.３６mmol)に、硫酸(４ml, ７５mmol)を加えた。反応物を４０℃で２８時間加熱した。反応物を室温まで冷却し、反応混合物がｐＨ 約７になるまで水性水酸化ナトリウム(約１５Ｍ)で反応停止させた。粗生成物を酢酸エチル(２×５０ml)で抽出し、有機物をＭｇＳＯ_４で乾燥させ、濾過し、真空で濃縮した。粗生成物をクロマトグラフィーによって精製し(Biotage, １００％ 酢酸エチルから９０／１０％ 酢酸エチル／ＭｅＯＨ)、７−クロロ−８−フルオロイソキノリン−３−アミンを粉末として得た(０.５８ｇ, ２.９５mmol, 収率８８％)。
¹H NMR (500 MHz, DMSO-d₆) d ppm 8.97 (s, 1 H), 7.47 - 7.57 (m, 1 H), 7.38 - 7.45 (m, 1 H), 6.67 (s, 1 H), 6.33 (s, 2 H). MS (LC/MS) R.T. = 1.18; [M+H]⁺ = 196.95。 Step B: 7-Chloro-8-fluoroisoquinolin-3-amine

To N- (3-chloro-2-fluorobenzyl) -2,2-diethoxyacetimidoamide (0.97 g, 3.36 mmol) was added sulfuric acid (4 ml, 75 mmol). The reaction was heated at 40 ° C. for 28 hours. The reaction was cooled to room temperature and quenched with aqueous sodium hydroxide (about 15M) until the reaction mixture was about pH 7. The crude product was extracted with ethyl acetate (2 × 50 ml) and the organics were dried over MgSO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography (Biotage, 100% ethyl acetate to 90/10% ethyl acetate / MeOH) to give 7-chloro-8-fluoroisoquinolin-3-amine as a powder (0.58 g, 2 .95 mmol, 88% yield).
¹ H NMR (500 MHz, DMSO-d ₆ ) d ppm 8.97 (s, 1 H), 7.47-7.57 (m, 1 H), 7.38-7.45 (m, 1 H), 6.67 (s, 1 H), 6.33 (s, 2 H). MS (LC / MS) RT = 1.18; [M + H] ⁺ = 196.95.

(Ｒ)−Ｎ−(７−クロロ−８−フルオロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６７の工程Ａ〜Ｂの一般的な手順に従って、７−クロロ−８−フルオロイソキノリン−３−アミン(上記の工程Ｂより)を出発物質として用いて製造した。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 9.19 (d, J=2.1 Hz, 1 H), 8.30 - 8.85 (m, 1 H), 7.68 (br. s., 2 H), 6.93 - 7.43 (m, 1 H), 3.73 - 4.10 (m, 1 H), 3.48 - 3.70 (m, 1 H), 3.02 (br. s., 2 H), 2.59 - 2.86 (m, 4 H), 1.80 - 2.08 (m, 2 H), 1.60 (br. s., 2 H), 1.37 - 1.53 (m, 1 H). MS (LC/MS) R.T. = 1.22; [M+H]⁺ = 361.06。 Step C: (R) -N- (7-chloro-8-fluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(R) -N- (7-chloro-8-fluoroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine Prepared according to the general procedure of Example 267, Steps AB, using 7-chloro-8-fluoroisoquinolin-3-amine (from Step B above) as the starting material.
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 9.19 (d, J = 2.1 Hz, 1 H), 8.30-8.85 (m, 1 H), 7.68 (br. S., 2 H), 6.93- 7.43 (m, 1 H), 3.73-4.10 (m, 1 H), 3.48-3.70 (m, 1 H), 3.02 (br. S., 2 H), 2.59-2.86 (m, 4 H), 1.80 -2.08 (m, 2 H), 1.60 (br. S., 2 H), 1.37-1.53 (m, 1 H). MS (LC / MS) RT = 1.22; [M + H] ⁺ = 361.06.

(Ｒ)−Ｎ−(１−メチルイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例３１２の工程Ａ〜Ｃの一般的な手順に従って、１−フェニルエタンアミンを出発物質として用いて製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.10 (d, J=8.2 Hz, 1 H), 7.72 (d, J=8.2 Hz, 1 H), 7.60 (t, J=7.5 Hz, 1 H), 7.44 (t, J=7.6 Hz, 1 H), 7.03 - 7.33 (m, 1 H), 4.01 (d, J=10.1 Hz, 1 H), 3.69 (d, J=10.1 Hz, 1 H), 3.19 - 3.31 (m, 2 H), 3.12 (d, J=15.0 Hz, 1 H), 2.90 - 3.01 (m, 5 H), 2.77 - 2.90 (m, 2 H), 2.17 (br. s., 2 H), 1.58 - 1.87 (m, 3 H). MS (LC/MS) R.T. = 1.09; [M+H]⁺ = 323.16。 Example 313
(R) -N- (1-Methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 312. Prepared according to the general procedure of Steps A to C using 1-phenylethanamine as the starting material.
¹ H NMR (500 MHz, MeOD) δ ppm 8.10 (d, J = 8.2 Hz, 1 H), 7.72 (d, J = 8.2 Hz, 1 H), 7.60 (t, J = 7.5 Hz, 1 H), 7.44 (t, J = 7.6 Hz, 1 H), 7.03-7.33 (m, 1 H), 4.01 (d, J = 10.1 Hz, 1 H), 3.69 (d, J = 10.1 Hz, 1 H), 3.19 -3.31 (m, 2 H), 3.12 (d, J = 15.0 Hz, 1 H), 2.90-3.01 (m, 5 H), 2.77-2.90 (m, 2 H), 2.17 (br. S., 2 H), 1.58-1.87 (m, 3 H). MS (LC / MS) RT = 1.09; [M + H] ⁺ = 323.16.

(Ｒ)−Ｎ−(６−(４−フルオロフェニル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２５６の工程Ａ〜Ｃの方法によって、６−クロロピリミジン−４−アミンおよび４−フルオロフェニルボロン酸から出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 8.75 (1 H, s), 7.93 - 8.11 (2 H, m), 7.06 - 7.30 (3 H, m), 4.03 (1 H, d, J=10.32 Hz), 3.73 (1 H, d, J=10.32 Hz), 3.18 - 3.27 (1 H, m), 3.05 - 3.16 (1 H, m), 2.66 - 3.00 (4 H, m), 2.01 - 2.20 (2 H, m), 1.49 - 1.87 (3 H, m). LC/MS RT=1.53; [M+H]⁺ = 354.24。 Example 314
(R) -N- (6- (4-Fluorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (4-Fluorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Was prepared by the method of Example 256, Steps AC, starting from 6-chloropyrimidin-4-amine and 4-fluorophenylboronic acid.
¹ H NMR (400 MHz, MeOD) δ ppm 8.75 (1 H, s), 7.93-8.11 (2 H, m), 7.06-7.30 (3 H, m), 4.03 (1 H, d, J = 10.32 Hz ), 3.73 (1 H, d, J = 10.32 Hz), 3.18-3.27 (1 H, m), 3.05-3.16 (1 H, m), 2.66-3.00 (4 H, m), 2.01-2.20 (2 H, m), 1.49-1.87 (3 H, m). LC / MS RT = 1.53; [M + H] ⁺ = 354.24.

(Ｒ)−Ｎ−(６−(３−フルオロフェニル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２５６の工程Ａ〜Ｃの方法によって、６−クロロピリミジン−４−アミンおよび３−フルオロフェニルボロン酸から出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 8.80 (1 H, d, J=1.26 Hz), 7.71 - 7.85 (2 H, m), 7.50 (1 H, td, J=7.99, 5.92 Hz), 7.11 - 7.36 (2 H, m), 4.08 (1 H, d, J=10.58 Hz), 3.80 (1 H, d, J=10.58 Hz), 3.39 - 3.48 (1 H, m), 3.33 (1 H, s), 2.83 - 3.20 (4 H, m), 2.10 - 2.33 (2 H, m), 1.65 - 1.96 (3 H, m). LC/MS RT=1.59; [M+H]⁺ = 354.24。 Example 315
(R) -N- (6- (3-Fluorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (3-Fluorophenyl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Was prepared by the method of Example 256, steps AC starting from 6-chloropyrimidin-4-amine and 3-fluorophenylboronic acid.
¹ H NMR (400 MHz, MeOD) δ ppm 8.80 (1 H, d, J = 1.26 Hz), 7.71-7.85 (2 H, m), 7.50 (1 H, td, J = 7.99, 5.92 Hz), 7.11 -7.36 (2 H, m), 4.08 (1 H, d, J = 10.58 Hz), 3.80 (1 H, d, J = 10.58 Hz), 3.39-3.48 (1 H, m), 3.33 (1 H, s), 2.83-3.20 (4 H, m), 2.10-2.33 (2 H, m), 1.65-1.96 (3 H, m). LC / MS RT = 1.59; [M + H] ⁺ = 354.24.

(Ｒ)−Ｎ−(５−フルオロピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３１の工程Ｂ〜Ｃの方法によって、２−クロロ−５−フルオロピリミジンから出発して製造した。
¹H NMR (400 MHz, MeOD)δ ppm 8.50 (2 H, d, J=0.76 Hz), 4.03 (1 H, d, J=10.07 Hz), 3.77 (1 H, d, J=10.32 Hz), 3.47 - 3.57 (1 H, m), 3.36 - 3.43 (1 H, m), 3.01 - 3.24 (4 H, m), 2.15 - 2.40 (2 H, m), 1.68 - 2.03 (3 H, m). LC/MS RT=0.30; [M+H]⁺ = 278.19。 Example 316
(R) -N- (5-Fluoropyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Fluoropyrimidin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 231. Prepared starting from 2-chloro-5-fluoropyrimidine by the method of steps B-C.
¹ H NMR (400 MHz, MeOD) δ ppm 8.50 (2 H, d, J = 0.76 Hz), 4.03 (1 H, d, J = 10.07 Hz), 3.77 (1 H, d, J = 10.32 Hz), 3.47-3.57 (1 H, m), 3.36-3.43 (1 H, m), 3.01-3.24 (4 H, m), 2.15-2.40 (2 H, m), 1.68-2.03 (3 H, m). LC / MS RT = 0.30; [M + H] ⁺ = 278.19.

(Ｒ)−Ｎ−(４−クロロ−５−メトキシピリミジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３１の工程Ｂ〜Ｃの方法によって、２,４−ジクロロ−５−メトキシピリミジンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 7.99 (1 H, s), 4.13 (1 H, d, J=10.83 Hz), 3.91 (4 H, d, J=10.83 Hz), 3.76 - 3.84 (1 H, m), 3.64 - 3.71 (1 H, m), 3.30 - 3.51 (4 H, m), 2.43 - 2.52 (1 H, m), 2.37 (1 H, d, J=3.27 Hz), 2.09 (1 H, dd, J=9.32, 4.78 Hz), 1.88 - 2.03 (2 H, m). LC/MS RT=0.30; [M+H]⁺ = 278.19。 Example 317
(R) -N- (4-Chloro-5-methoxypyrimidin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (4-Chloro-5-methoxypyrimidin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared starting from 2,4-dichloro-5-methoxypyrimidine, by the method of Example 231, Steps BC.
¹ H NMR (400 MHz, MeOD) δ ppm 7.99 (1 H, s), 4.13 (1 H, d, J = 10.83 Hz), 3.91 (4 H, d, J = 10.83 Hz), 3.76-3.84 (1 H, m), 3.64-3.71 (1 H, m), 3.30-3.51 (4 H, m), 2.43-2.52 (1 H, m), 2.37 (1 H, d, J = 3.27 Hz), 2.09 ( 1 H, dd, J = 9.32, 4.78 Hz), 1.88-2.03 (2 H, m). LC / MS RT = 0.30; [M + H] ⁺ = 278.19.

(Ｒ)−Ｎ−(６,８−ジフルオロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、(３,５−ジフルオロフェニル)メタンアミンから出発して製造した。
MS (LC/MS) R.T. = 1.42; [M+H]⁺ = 345.26。 Example 318
(R) -N- (6,8-difluoroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6,8-difluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 279, steps AD, starting from (3,5-difluorophenyl) methanamine.
MS (LC / MS) RT = 1.42; [M + H] ⁺ = 345.26.

(Ｒ)−Ｎ−(６,７−ジフルオロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、(３,４−ジフルオロフェニル)メタンアミンから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.97 (1 H, s), 7.73 - 7.91 (1 H, m), 7.58 (1 H, dd, J=11.14, 7.78 Hz), 7.30 (1 H, s), 4.01 (1 H, d, J=10.07 Hz), 3.71 (1 H, d, J=10.07 Hz), 3.31 (1 H, br. s.), 3.14 - 3.24 (1 H, m), 3.03 (2 H, d, J=7.32 Hz), 2.83 - 2.98 (2 H, m), 2.20 (2 H, br. s.), 1.61 - 1.91 (3 H, m). MS (LC/MS) R.T. = 1.45; [M+H]⁺ = 345.26。 Example 319
(R) -N- (6,7-difluoroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6,7-difluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine is carried out Prepared by the method of Example 279, steps AD, starting from (3,4-difluorophenyl) methanamine.
¹ H NMR (500 MHz, MeOD) δ ppm 8.97 (1 H, s), 7.73-7.91 (1 H, m), 7.58 (1 H, dd, J = 11.14, 7.78 Hz), 7.30 (1 H, s ), 4.01 (1 H, d, J = 10.07 Hz), 3.71 (1 H, d, J = 10.07 Hz), 3.31 (1 H, br.s.), 3.14-3.24 (1 H, m), 3.03 (2 H, d, J = 7.32 Hz), 2.83-2.98 (2 H, m), 2.20 (2 H, br. S.), 1.61-1.91 (3 H, m). MS (LC / MS) RT = 1.45; [M + H] ⁺ = 345.26.

(Ｒ)−Ｎ−(５,８−ジフルオロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、(２,５−ジフルオロフェニル)メタンアミンから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 9.25 (1 H, s), 7.43 (1 H, br. s.), 7.20 - 7.34 (1 H, m), 6.91 - 7.12 (1 H, m), 4.04 (1 H, d, J=10.07 Hz), 3.74 (1 H, d, J=10.07 Hz), 3.30 (1 H, br. s.), 3.13 - 3.23 (1 H, m), 3.03 (2 H, d, J=7.63 Hz), 2.82 - 2.96 (2 H, m), 2.21 (2 H, br. s.), 1.58 - 1.92 (3 H, m). MS (LC/MS) R.T. = 1.37; [M+H]⁺ = 345.33。 Example 320
(R) -N- (5,8-difluoroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5,8-difluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine is carried out Prepared by the method of Example 279, steps AD, starting from (2,5-difluorophenyl) methanamine.
¹ H NMR (500 MHz, MeOD) δ ppm 9.25 (1 H, s), 7.43 (1 H, br. S.), 7.20-7.34 (1 H, m), 6.91-7.12 (1 H, m), 4.04 (1 H, d, J = 10.07 Hz), 3.74 (1 H, d, J = 10.07 Hz), 3.30 (1 H, br. S.), 3.13-3.23 (1 H, m), 3.03 (2 H, d, J = 7.63 Hz), 2.82-2.96 (2 H, m), 2.21 (2 H, br. S.), 1.58-1.92 (3 H, m). MS (LC / MS) RT = 1.37 ; [M + H] ⁺ = 345.33.

(Ｒ)−Ｎ−(７−フルオロキナゾリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３９の工程Ａ〜Ｂの方法によって、７−フルオロキナゾリン−２−アミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.22 (1 H, s), 7.95 (1 H, dd, J=8.81, 6.04 Hz), 7.47 (1 H, dd, J=10.45, 2.14 Hz), 7.24 (1 H, td, J=8.75, 2.39 Hz), 4.07 (1 H, d, J=10.07 Hz), 3.77 (1 H, d, J=10.07 Hz), 3.25 (1 H, s), 3.07 - 3.17 (1 H, m), 2.89 - 3.02 (2 H, m), 2.75 - 2.88 (2 H, m), 2.04 - 2.26 (2 H, m), 1.50 - 1.85 (3 H, m). MS (LC/MS) R.T. =1.06; [M+H]⁺= 328.33。 Example 321
(R) -N- (7-Fluoroquinazolin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (7-Fluoroquinazolin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 239. Prepared starting from 7-fluoroquinazolin-2-amine by the method of Steps AB.
¹ H NMR (400 MHz, MeOD) δ ppm 9.22 (1 H, s), 7.95 (1 H, dd, J = 8.81, 6.04 Hz), 7.47 (1 H, dd, J = 10.45, 2.14 Hz), 7.24 (1 H, td, J = 8.75, 2.39 Hz), 4.07 (1 H, d, J = 10.07 Hz), 3.77 (1 H, d, J = 10.07 Hz), 3.25 (1 H, s), 3.07- 3.17 (1 H, m), 2.89-3.02 (2 H, m), 2.75-2.88 (2 H, m), 2.04-2.26 (2 H, m), 1.50-1.85 (3 H, m). MS ( LC / MS) RT = 1.06; [M + H] ⁺ = 328.33.

(Ｒ)−Ｎ−(５−クロロキナゾリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３９の工程Ａ〜Ｂの方法によって、５−クロロキナゾリン−２−アミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.53 (1 H, s), 7.62 - 7.84 (2 H, m), 7.47 (1 H, dd, J=6.80, 1.51 Hz), 4.09 (1 H, d, J=10.32 Hz), 3.80 (1 H, d, J=10.32 Hz), 3.36 (1 H, s), 3.15 - 3.24 (1 H, m), 3.00 (2 H, t, J=7.68 Hz), 2.79 - 2.95 (2 H, m), 2.05 - 2.32 (2 H, m), 1.53 - 1.87 (3 H, m). MS (LC/MS) R.T. =1.36; [M+H]⁺= 344.29。 Example 322
(R) -N- (5-chloroquinazolin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Chloroquinazolin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared according to Example 239. Prepared starting from 5-chloroquinazolin-2-amine by the method of Steps AB.
¹ H NMR (400 MHz, MeOD) δ ppm 9.53 (1 H, s), 7.62-7.84 (2 H, m), 7.47 (1 H, dd, J = 6.80, 1.51 Hz), 4.09 (1 H, d , J = 10.32 Hz), 3.80 (1 H, d, J = 10.32 Hz), 3.36 (1 H, s), 3.15-3.24 (1 H, m), 3.00 (2 H, t, J = 7.68 Hz) , 2.79-2.95 (2 H, m), 2.05-2.32 (2 H, m), 1.53-1.87 (3 H, m). MS (LC / MS) RT = 1.36; [M + H] ⁺ = 344.29.

(Ｒ)−Ｎ−(５−フルオロキナゾリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３９の工程Ａ〜Ｂの方法によって、５−フルオロキナゾリン−２−アミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.46 (1 H, s), 7.77 (1 H, td, J=8.18, 6.30 Hz), 7.63 (1 H, d, J=8.56 Hz), 7.00 - 7.20 (1 H, m), 4.08 (1 H, d, J=10.07 Hz), 3.77 (1 H, d, J=10.07 Hz), 3.25 (1 H, s), 3.07 - 3.17 (1 H, m), 2.94 (2 H, t, J=7.68 Hz), 2.72 - 2.88 (2 H, m), 2.05 - 2.21 (2 H, m), 1.48 - 1.83 (3 H, m). MS (LC/MS) R.T. =1.10; [M+H]⁺= 328.33。 Example 323
(R) -N- (5-Fluoroquinazolin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (5-Fluoroquinazolin-2-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared in Example 239. Prepared starting from 5-fluoroquinazolin-2-amine by the method of Steps AB.
¹ H NMR (400 MHz, MeOD) δ ppm 9.46 (1 H, s), 7.77 (1 H, td, J = 8.18, 6.30 Hz), 7.63 (1 H, d, J = 8.56 Hz), 7.00-7.20 (1 H, m), 4.08 (1 H, d, J = 10.07 Hz), 3.77 (1 H, d, J = 10.07 Hz), 3.25 (1 H, s), 3.07-3.17 (1 H, m) , 2.94 (2 H, t, J = 7.68 Hz), 2.72-2.88 (2 H, m), 2.05-2.21 (2 H, m), 1.48-1.83 (3 H, m). MS (LC / MS) RT = 1.10; [M + H] ⁺ = 328.33.

(Ｒ)−Ｎ−(７−フルオロ−４−メチルキナゾリン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３９の工程Ａ〜Ｂの方法によって、７−フルオロ−４−メチルキナゾリン−２−アミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, dd, J=9.07, 6.04 Hz), 7.46 (1 H, dd, J=10.45, 2.64 Hz), 7.22 (1 H, td, J=8.81, 2.77 Hz), 4.06 (1 H, d, J=10.07 Hz), 3.75 (1 H, d, J=10.07 Hz), 3.23 (1 H, s), 3.06 - 3.15 (1 H, m), 2.94 (2 H, t, J=7.68 Hz), 2.73 - 2.88 (5 H, m), 2.06 - 2.24 (2 H, m), 1.51 - 1.86 (3 H, m). MS (LC/MS) R.T. =1.25; [M+H]⁺= 342.29。 Example 324
(R) -N- (7-Fluoro-4-methylquinazolin-2-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(R) -N- (7-Fluoro-4-methylquinazolin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 239, steps AB, starting from 7-fluoro-4-methylquinazolin-2-amine.
¹ H NMR (400 MHz, MeOD) δ ppm 8.13 (1 H, dd, J = 9.07, 6.04 Hz), 7.46 (1 H, dd, J = 10.45, 2.64 Hz), 7.22 (1 H, td, J = 8.81, 2.77 Hz), 4.06 (1 H, d, J = 10.07 Hz), 3.75 (1 H, d, J = 10.07 Hz), 3.23 (1 H, s), 3.06-3.15 (1 H, m), 2.94 (2 H, t, J = 7.68 Hz), 2.73-2.88 (5 H, m), 2.06-2.24 (2 H, m), 1.51-1.86 (3 H, m). MS (LC / MS) RT = 1.25; [M + H] ⁺ = 342.29.

(Ｒ)−Ｎ−(６−フルオロ−１−メチルイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、(Ｓ)−１−(４−フルオロフェニル)エタンアミンから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.23 (1 H, dd, J=9.16, 5.49 Hz), 7.44 (1 H, dd, J=9.92, 2.59 Hz), 7.22 - 7.38 (2 H, m), 4.17 (1 H, d, J=10.99 Hz), 3.94 (1 H, d, J=10.68 Hz), 3.71 - 3.79 (1 H, m), 3.61 - 3.69 (1 H, m), 3.39 - 3.49 (1 H, m), 3.33 - 3.39 (3 H, m), 2.95 (3 H, s), 2.49 (1 H, br. s.), 2.37 (1 H, tt, J=10.07, 3.36 Hz), 2.05 - 2.16 (1 H, m), 1.93 - 2.05 (2 H, m). MS (LC/MS) R.T. =1.47; [M+H]⁺= 341.25。 Example 325
(R) -N- (6-Fluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Fluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 279, steps AD, starting from (S) -1- (4-fluorophenyl) ethanamine.
¹ H NMR (500 MHz, MeOD) δ ppm 8.23 (1 H, dd, J = 9.16, 5.49 Hz), 7.44 (1 H, dd, J = 9.92, 2.59 Hz), 7.22-7.38 (2 H, m) , 4.17 (1 H, d, J = 10.99 Hz), 3.94 (1 H, d, J = 10.68 Hz), 3.71-3.79 (1 H, m), 3.61-3.69 (1 H, m), 3.39-3.49 (1 H, m), 3.33-3.39 (3 H, m), 2.95 (3 H, s), 2.49 (1 H, br. S.), 2.37 (1 H, tt, J = 10.07, 3.36 Hz) , 2.05-2.16 (1 H, m), 1.93-2.05 (2 H, m). MS (LC / MS) RT = 1.47; [M + H] ⁺ = 341.25.

(Ｒ)−Ｎ−(６−クロロ−１−メチルイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、１−(４−クロロフェニル)エタンアミンから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.23 (1 H, dd, J=9.16, 5.49 Hz), 7.44 (1 H, dd, J=9.92, 2.59 Hz), 7.22 - 7.38 (2 H, m), 4.17 (1 H, d, J=10.99 Hz), 3.94 (1 H, d, J=10.68 Hz), 3.71 - 3.79 (1 H, m), 3.61 - 3.69 (1 H, m), 3.39 - 3.49 (1 H, m), 3.33 - 3.39 (3 H, m), 2.95 (3 H, s), 2.49 (1 H, br. s.), 2.37 (1 H, tt, J=10.07, 3.36 Hz), 2.05 - 2.16 (1 H, m), 1.93 - 2.05 (2 H, m). MS (LC/MS) R.T. =1.79; [M+H]⁺= 357.28。 Example 326
(R) -N- (6-Chloro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6-Chloro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 279, steps AD, starting from 1- (4-chlorophenyl) ethanamine.
¹ H NMR (500 MHz, MeOD) δ ppm 8.23 (1 H, dd, J = 9.16, 5.49 Hz), 7.44 (1 H, dd, J = 9.92, 2.59 Hz), 7.22-7.38 (2 H, m) , 4.17 (1 H, d, J = 10.99 Hz), 3.94 (1 H, d, J = 10.68 Hz), 3.71-3.79 (1 H, m), 3.61-3.69 (1 H, m), 3.39-3.49 (1 H, m), 3.33-3.39 (3 H, m), 2.95 (3 H, s), 2.49 (1 H, br. S.), 2.37 (1 H, tt, J = 10.07, 3.36 Hz) , 2.05-2.16 (1 H, m), 1.93-2.05 (2 H, m). MS (LC / MS) RT = 1.79; [M + H] ⁺ = 357.28.

(Ｒ)−Ｎ−(７−フルオロ−１−メチルイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、１−(３−フルオロフェニル)エタンアミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 7.55 - 7.81 (2 H, m), 7.38 (1 H, t, J=7.55 Hz), 7.16 (1 H, br. s.), 3.95 (1 H, d, J=9.82 Hz), 3.64 (1 H, d, J=9.82 Hz), 3.20 (1 H, d, J=14.86 Hz), 3.01 - 3.13 (1 H, m), 2.63 - 2.97 (7 H, m), 2.11 (2 H, br. s.), 1.41 - 1.81 (3 H, m). MS (LC/MS) R.T. =1.63; [M+H]⁺= 341.32。 Example 327
(R) -N- (7-Fluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (7-Fluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine Prepared by the method of Example 279, steps AD, starting from 1- (3-fluorophenyl) ethanamine.
¹ H NMR (400 MHz, MeOD) δ ppm 7.55-7.81 (2 H, m), 7.38 (1 H, t, J = 7.55 Hz), 7.16 (1 H, br.s.), 3.95 (1 H, d, J = 9.82 Hz), 3.64 (1 H, d, J = 9.82 Hz), 3.20 (1 H, d, J = 14.86 Hz), 3.01-3.13 (1 H, m), 2.63-2.97 (7 H , m), 2.11 (2 H, br. s.), 1.41-1.81 (3 H, m). MS (LC / MS) RT = 1.63; [M + H] ⁺ = 341.32.

(Ｒ)−Ｎ−(５,７−ジフルオロ−１−メチルイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２７９の工程Ａ〜Ｄの方法によって、１−(３,５−ジフルオロフェニル)エタンアミンから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 7.78 (1 H, d, J=9.77 Hz), 7.39 - 7.55 (2 H, m), 4.31 (1 H, d, J=10.99 Hz), 4.11 (1 H, d, J=10.99 Hz), 3.88 - 3.97 (1 H, m), 3.79 - 3.86 (1 H, m), 3.55 (1 H, t, J=11.90 Hz), 3.37 - 3.47 (3 H, m), 3.00 (3 H, s), 2.64 (1 H, br. s.), 2.32 - 2.45 (1 H, m), 2.18 (1 H, dddd, J=11.71, 7.13, 6.94, 6.56 Hz), 1.99 - 2.13 (2 H, m). MS (LC/MS) R.T. =1.67; [M+H]⁺= 359.22。 Example 328
(R) -N- (5,7-Difluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

(R) -N- (5,7-Difluoro-1-methylisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- The amine was prepared by the method of Example 279, steps AD, starting from 1- (3,5-difluorophenyl) ethanamine.
¹ H NMR (500 MHz, MeOD) δ ppm 7.78 (1 H, d, J = 9.77 Hz), 7.39-7.55 (2 H, m), 4.31 (1 H, d, J = 10.99 Hz), 4.11 (1 H, d, J = 10.99 Hz), 3.88-3.97 (1 H, m), 3.79-3.86 (1 H, m), 3.55 (1 H, t, J = 11.90 Hz), 3.37-3.47 (3 H, m), 3.00 (3 H, s), 2.64 (1 H, br.s.), 2.32-2.45 (1 H, m), 2.18 (1 H, dddd, J = 11.71, 7.13, 6.94, 6.56 Hz) , 1.99-2.13 (2 H, m). MS (LC / MS) RT = 1.67; [M + H] ⁺ = 359.22.

(Ｒ)−Ｎ−(６−(６−(メチルチオ)ピリジン−３−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２６１の工程Ａ〜Ｂの方法によって、６−(６−フルオロピリジン−３−イル)ピリミジン−４−アミンから出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.02 (1 H, d, J=2.52 Hz), 8.78 (1 H, s), 8.20 (1 H, dd, J=8.44, 2.39 Hz), 7.36 (1 H, d, J=8.56 Hz), 7.26 (1 H, br. s.), 4.06 (1 H, d, J=10.58 Hz), 3.77 (1 H, d, J=10.32 Hz), 3.34 (1 H, s), 3.14 - 3.22 (1 H, m), 2.82 - 3.08 (4 H, m), 2.58 (3 H, s), 2.08 - 2.24 (2 H, m), 1.60 - 1.88 (3 H, m). MS (LC/MS) R.T. =3.09; [M+H]⁺= 383.2。 Example 329
(R) -N- (6- (6- (6- (Methylthio) pyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine

(R) -N- (6- (6- (6- (Methylthio) pyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine was prepared by the method of Example 261, Steps AB, starting from 6- (6-fluoropyridin-3-yl) pyrimidin-4-amine.
¹ H NMR (400 MHz, MeOD) δ ppm 9.02 (1 H, d, J = 2.52 Hz), 8.78 (1 H, s), 8.20 (1 H, dd, J = 8.44, 2.39 Hz), 7.36 (1 H, d, J = 8.56 Hz), 7.26 (1 H, br.s.), 4.06 (1 H, d, J = 10.58 Hz), 3.77 (1 H, d, J = 10.32 Hz), 3.34 (1 H, s), 3.14-3.22 (1 H, m), 2.82-3.08 (4 H, m), 2.58 (3 H, s), 2.08-2.24 (2 H, m), 1.60-1.88 (3 H, m). MS (LC / MS) RT = 3.09; [M + H] ⁺ = 383.2.

Example 330
(R) -N- (6- (2-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

６−クロロピリミジン−４−アミン(１.５ｇ, １１.５８mmol)、２−フルオロピリジン−３−イルボロン酸(２.０３９ｇ, １４.４７mmol)およびＮａ_２ＣＯ_３(３.６８ｇ, ３４.７mmol)の混合物を、ジオキサン(１５ml)／ＥｔＯＨ(２ml)／水(３ml)の混合物に懸濁した。混合物を、マイクロ波合成機中で、１２５℃で２０分間加熱し、濃縮し、シリカゲルのカートリッジで、ヘキサン中１０〜６０％ 酢酸エチル、次に酢酸エチル中５〜２５％ ９：１ メタノール：水酸化アンモニウムを用いて精製し、灰白色の固体を得た(１.５ｇ, ７.８９mmol, ６８％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.57 (1 H, ddd, J=9.95, 7.68, 2.01 Hz), 8.48 (1 H, d, J=1.26 Hz), 8.33 (1 H, td, J=3.02, 2.01 Hz), 7.51 (1 H, ddd, J=7.37, 4.97, 2.01 Hz), 7.12 (2 H, br. s.), 6.96 (1 H, d, J=1.26 Hz). MS (LC/MS) R.T. =0.43; [M+H]⁺= 191.15。 Step A: 6- (2-Fluoropyridin-3-yl) pyrimidin-4-amine

6-chloropyrimidin-4-amine (1.5 g, 11.58 mmol), 2-fluoropyridin-3-ylboronic acid (2.039 g, 14.47 mmol) and Na ₂ CO 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 a microwave synthesizer at 125 ° C. for 20 minutes, concentrated and silica gel cartridge with 10-60% ethyl acetate in hexane, then 5-25% in ethyl acetate 9: 1 methanol: water. Purification using ammonium oxide gave an off-white solid (1.5 g, 7.89 mmol, 68%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.57 (1 H, ddd, J = 9.95, 7.68, 2.01 Hz), 8.48 (1 H, d, J = 1.26 Hz), 8.33 (1 H, td , J = 3.02, 2.01 Hz), 7.51 (1 H, ddd, J = 7.37, 4.97, 2.01 Hz), 7.12 (2 H, br.s.), 6.96 (1 H, d, J = 1.26 Hz). MS (LC / MS) RT = 0.43; [M + H] ⁺ = 191.15.

ＤＭＦ中の６−(２−フルオロピリジン−３−イル)ピリミジン−４−アミン(０.３ｇ, １.５７７mmol)の溶液に、ＮａＨ(０.１２６ｇ, ３.１５mmol)を加えた。１.５時間撹拌を続け、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.３６６ｇ, １.５７７mmol)を加えた。混合物を室温で２時間撹拌し、シリカゲルで、ヘキサン中１０〜１５％ 酢酸エチルを用いて精製し、橙色の固体を得た(０.１３３ｇ, ０.５７３mmol, ３６％)。
MS (LC/MS) R.T. = 2.48; [M+H]⁺= 233.08。 Step B: 4- (2-Fluoropyridin-3-yl) -6-isothiocyanatopyrimidine

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 hours and 1,1′-thiocarbonyldipyridin-2 (1H) -one (0.366 g, 1.577 mmol) was added. The mixture was stirred at room temperature for 2 hours and purified on silica gel with 10-15% ethyl acetate in hexanes to give an orange solid (0.133 g, 0.573 mmol, 36%).
MS (LC / MS) RT = 2.48; [M + H] ⁺ = 233.08.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の４−(２−フルオロピリジン−３−イル)−６−イソチオシアナトピリミジン(０.１８ｇ, ０.７７５mmol)に、Ｃｓ_２ＣＯ_３(０.６３１ｇ, １.９３８mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１７８ｇ, ０.７７５mmol)を加えた。懸濁液を室温で１８時間撹拌し、濃縮した。残渣を、シリカゲルで、酢酸エチル中０〜１０％ ９.５：０.５ メタノール：水酸化アンモニウムを用いて精製し、黄色の固体を得た(０.２ｇ, ０.５１５mmol, ６６％)。
MS (LC/MS) R.T. = 1.82; [M+H]⁺= 389.27。 Step C: (S) -1- (6- (2-Fluoropyridin-3-yl) pyrimidin-4-yl) -3-((3-hydroxyquinuclidin-3-yl) methyl) thiourea

4- (2-Fluoropyridin-3-yl) -6-isothiocyanatopyrimidine (0.18 g, 0.775 mmol) in N, N-dimethylformamide (20 ml) was added to Cs ₂ CO ₃ (0.631 g, 1.938 mmol) and (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (0.178 g, 0.775 mmol) were added. The suspension was stirred at room temperature for 18 hours and concentrated. The residue was purified on silica gel using 0-10% in ethyl acetate 9.5: 0.5 methanol: ammonium hydroxide to give a yellow solid (0.2 g, 0.515 mmol, 66%).
MS (LC / MS) RT = 1.82; [M + H] ⁺ = 389.27.

Ｎ,Ｎ−ジメチルホルムアミド(２０ml)中の(Ｓ)−１−(６−(２−フルオロピリジン−３−イル)ピリミジン−４−イル)−３−((３−ヒドロキシキヌクリジン−３−イル)メチル)チオウレア(０.２ｇ, ０.５１５mmol)の懸濁液に、Ｃｓ_２ＣＯ_３(０.１６８ｇ, ０.５１５mmol)およびＤＩＣ(０.２４１ml, １.５４５mmol)を加えた。懸濁液を、室温で１８時間、次に６０℃で３時間撹拌した。混合物を濃縮し、残渣を、シリカゲルで、酢酸エチル中２〜７％ ９.５：０.５ メタノール：水酸化アンモニウムを用いて精製し、(Ｒ)−Ｎ−(６−(２−フルオロピリジン−３−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを薄黄色の固体として得た(０.０２９ｇ, ０.０８１mmol, １６％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.83 (1 H, d, J=1.26 Hz), 8.55 (1 H, ddd, J=9.76, 7.62, 2.01 Hz), 8.29 (1 H, dt, J=3.02, 1.51 Hz), 7.46 (1 H, ddd, J=7.37, 5.10, 1.89 Hz), 7.33 (1 H, s), 4.06 (1 H, d, J=10.32 Hz), 3.76 (1 H, d, J=10.32 Hz), 3.25 (1 H, s), 3.09 - 3.19 (1 H, m), 2.95 (2 H, t, J=7.68 Hz), 2.74 - 2.91 (2 H, m), 2.04 - 2.26 (2 H, m), 1.55 - 1.87 (3 H, m). MS (LC/MS) R.T. = 1.24; [M+H]⁺= 355.28。 Step D: (R) -N- (6- (2-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine)

(S) -1- (6- (2-Fluoropyridin-3-yl) pyrimidin-4-yl) -3-((3-hydroxyquinuclidine-3-) in N, N-dimethylformamide (20 ml) To a suspension of (yl) methyl) thiourea (0.2 g, 0.515 mmol) was added Cs ₂ CO ₃ (0.168 g, 0.515 mmol) and DIC (0.241 ml, 1.545 mmol). The suspension was stirred at room temperature for 18 hours and then at 60 ° C. for 3 hours. The mixture is concentrated and the residue is purified on silica gel using 2-7% in ethyl acetate 9.5: 0.5 methanol: ammonium hydroxide and (R) -N- (6- (2-fluoropyridine). -3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a pale yellow solid (0 0.029 g, 0.081 mmol, 16%).
¹ H NMR (400 MHz, MeOD) δ ppm 8.83 (1 H, d, J = 1.26 Hz), 8.55 (1 H, ddd, J = 9.76, 7.62, 2.01 Hz), 8.29 (1 H, dt, J = 3.02, 1.51 Hz), 7.46 (1 H, ddd, J = 7.37, 5.10, 1.89 Hz), 7.33 (1 H, s), 4.06 (1 H, d, J = 10.32 Hz), 3.76 (1 H, d , J = 10.32 Hz), 3.25 (1 H, s), 3.09-3.19 (1 H, m), 2.95 (2 H, t, J = 7.68 Hz), 2.74-2.91 (2 H, m), 2.04- 2.26 (2 H, m), 1.55-1.87 (3 H, m). MS (LC / MS) RT = 1.24; [M + H] ⁺ = 355.28.

(Ｒ)−Ｎ−(６−(６−フルオロピリジン−３−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例３３０の工程Ａ〜Ｄの方法によって、６−フルオロピリジン−３−イルボロン酸から出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 8.88 (1 H, d, J=2.52 Hz), 8.82 (1 H, s), 8.51 - 8.60 (1 H, m), 7.25 (1 H, s), 7.19 (1 H, dd, J=8.56, 2.52 Hz), 4.06 (1 H, d, J=10.32 Hz), 3.76 (1 H, d, J=10.32 Hz), 3.22 (1 H, s), 3.07 - 3.14 (1 H, m), 2.86 - 2.98 (2 H, m), 2.70 - 2.87 (2 H, m), 2.03 - 2.20 (2 H, m), 1.54 - 1.87 (3 H, m). MS (LC/MS) R.T. = 1.21; [M+H]⁺= 355.28。 Example 331
(R) -N- (6- (6-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (6-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] The 2-amine was prepared by the method of Example 330, steps AD starting from 6-fluoropyridin-3-ylboronic acid.
¹ H NMR (400 MHz, MeOD) δ ppm 8.88 (1 H, d, J = 2.52 Hz), 8.82 (1 H, s), 8.51-8.60 (1 H, m), 7.25 (1 H, s), 7.19 (1 H, dd, J = 8.56, 2.52 Hz), 4.06 (1 H, d, J = 10.32 Hz), 3.76 (1 H, d, J = 10.32 Hz), 3.22 (1 H, s), 3.07 -3.14 (1 H, m), 2.86-2.98 (2 H, m), 2.70-2.87 (2 H, m), 2.03-2.20 (2 H, m), 1.54-1.87 (3 H, m). MS (LC / MS) RT = 1.21; [M + H] ⁺ = 355.28.

(Ｒ)−Ｎ−(６−(５−フルオロピリジン−３−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例３３０の工程Ａ〜Ｄの方法によって、５−フルオロピリジン−３−イルボロン酸から出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.06 (1 H, d, J=1.26 Hz), 8.81 (1 H, d, J=1.01 Hz), 8.56 (1 H, d, J=2.77 Hz), 8.26 (1 H, dt, J=9.82, 2.27 Hz), 7.30 (1 H, br. s.), 4.06 (1 H, d, J=10.32 Hz), 3.75 (1 H, d, J=10.32 Hz), 3.16 - 3.26 (1 H, m), 3.04 - 3.15 (1 H, m), 2.86 - 3.00 (2 H, m), 2.81 (2 H, t, J=7.30 Hz), 2.01 - 2.22 (2 H, m), 1.52 - 1.84 (3 H, m). MS (LC/MS) R.T. = 1.20; [M+H]⁺= 355.28。 Example 332
(R) -N- (6- (5-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(R) -N- (6- (5-Fluoropyridin-3-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] The 2-amine was prepared by the method of Example 330, steps AD starting from 5-fluoropyridin-3-ylboronic acid.
¹ H NMR (400 MHz, MeOD) δ ppm 9.06 (1 H, d, J = 1.26 Hz), 8.81 (1 H, d, J = 1.01 Hz), 8.56 (1 H, d, J = 2.77 Hz), 8.26 (1 H, dt, J = 9.82, 2.27 Hz), 7.30 (1 H, br.s.), 4.06 (1 H, d, J = 10.32 Hz), 3.75 (1 H, d, J = 10.32 Hz) ), 3.16-3.26 (1 H, m), 3.04-3.15 (1 H, m), 2.86-3.00 (2 H, m), 2.81 (2 H, t, J = 7.30 Hz), 2.01-2.22 (2 H, m), 1.52-1.84 (3 H, m). MS (LC / MS) RT = 1.20; [M + H] ⁺ = 355.28.

((Ｒ)−Ｎ−(６−(１−メチル−１Ｈ−ピラゾール−５−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例３３０の工程Ａ〜Ｄの方法によって、１−メチル−５−(４,４,５,５−テトラメチル−１,３,２−ジオキサボロラン−２−イル)−１Ｈ−ピラゾールから出発して製造した。
¹H NMR (500 MHz, MeOD) δ ppm 8.89 (1 H, s), 7.54 (1 H, d, J=2.14 Hz), 7.28 (1 H, br. s.), 6.84 (1 H, d, J=2.14 Hz), 4.23 (3 H, s), 4.18 (1 H, d, J=10.99 Hz), 3.98 (1 H, d, J=10.68 Hz), 3.81 - 3.88 (1 H, m), 3.72 - 3.80 (1 H, m), 3.53 (1 H, t, J=11.75 Hz), 3.34 - 3.46 (3 H, m), 2.53 (1 H, br. s.), 2.33 - 2.45 (1 H, m), 2.10 - 2.22 (1 H, m), 1.97 - 2.12 (2 H, m). MS (LC/MS) R.T. = 1.15; [M+H]⁺= 340.29。 Example 333
(R) -N- (6- (1-Methyl-1H-pyrazol-5-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2 ] Octane] -2-amine

((R) -N- (6- (1-Methyl-1H-pyrazol-5-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2. 2] Octane] -2-amine was prepared by 1-methyl-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- Yl) -1H-pyrazole.
¹ H NMR (500 MHz, MeOD) δ ppm 8.89 (1 H, s), 7.54 (1 H, d, J = 2.14 Hz), 7.28 (1 H, br.s.), 6.84 (1 H, d, J = 2.14 Hz), 4.23 (3 H, s), 4.18 (1 H, d, J = 10.99 Hz), 3.98 (1 H, d, J = 10.68 Hz), 3.81-3.88 (1 H, m), 3.72-3.80 (1 H, m), 3.53 (1 H, t, J = 11.75 Hz), 3.34-3.46 (3 H, m), 2.53 (1 H, br.s.), 2.33-2.45 (1 H , m), 2.10-2.22 (1 H, m), 1.97-2.12 (2 H, m). MS (LC / MS) RT = 1.15; [M + H] ⁺ = 340.29.

Example 334
(R) -N- (6- (pyrazin-2-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2- Amine

６−クロロピリミジン−４−アミン(４.１ｇ, ３１.６mmol)をアセトニトリルに懸濁し、ＤＭＡＰ(０.９６６ｇ, ７.９１mmol)およびジ−ｔｅｒｔ−ブチル ジカーボネート(１４.７４ｇ, ６７.５mmol)を加え、混合物を室温で３日間撹拌した。溶媒を蒸発させ、残渣を、シリカゲルによって、ヘキサン中０〜１５％ 酢酸エチルを用いて精製し、６−[ビス(ｔｅｒｔ−ブトキシカルボニル)アミノ]−４−クロロピリミジンを白色の固体として得た(６.２７ｇ, １９.０１mmol, ６０％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.69 (1 H, d, J=0.76 Hz), 7.88 (1 H, d, J=1.01 Hz), 1.58 (18 H, s)。 Step A: 6- [Bis (tert-butoxycarbonyl) amino] -4-chloropyrimidine

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). And the mixture was stirred at room temperature for 3 days. The solvent was evaporated and the residue was purified by silica gel with 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%).
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.69 (1 H, d, J = 0.76 Hz), 7.88 (1 H, d, J = 1.01 Hz), 1.58 (18 H, s).

６−[ビス(ｔｅｒｔ−ブトキシカルボニル)アミノ]−４−クロロピリミジン(１.６０８ｇ, ４.８８mmol)、２−(トリブチルスタンニル)ピラジン(１.５ｇ, ４.０６mmol)およびＰｄ(Ｐｈ_３Ｐ)_４(０.２２５ｇ, ０.１９５mmol)を、トルエン中で合わせて、窒素を吹きつけ、１７時間還流し、室温まで冷却し、濃縮し、シリカゲルで、ヘキサン中１０％ 酢酸エチル、次にヘキサン中２０〜５０％ 酢酸エチルを用いて精製し、黄色の固体を得た(０.９４ｇ, ２.５０７mmol, ６２％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.67 (1 H, d, J=1.01 Hz), 9.05 (1 H, d, J=1.26 Hz), 8.60 - 8.77 (3 H, m), 1.59 (18 H, s)。 Step B: 6- [Bis (tert-butoxycarbonyl) amino] -4- (2′-pyrazinyl) pyrimidine

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 ₃ P ) ₄ (0.225 g, 0.195 mmol) were combined in toluene, blown with nitrogen, refluxed for 17 hours, cooled to room temperature, concentrated, and silica gel with 10% ethyl acetate in hexane, then hexane. Purification with medium 20-50% ethyl acetate gave a yellow solid (0.94 g, 2.507 mmol, 62%).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.67 (1 H, d, J = 1.01 Hz), 9.05 (1 H, d, J = 1.26 Hz), 8.60-8.77 (3 H, m), 1.59 (18 H, s).

トリフルオロ酢酸(２.７７ml, ３６.０mmol)を、ジクロロメタン中の６−[ビス(ｔｅｒｔ−ブトキシカルボニル)アミノ]−４−(２'−ピラジニル)ピリミジン(１.３４４ｇ, ３.６０mmol)の溶液に室温で加えた。混合物を室温で１８時間撹拌し、濃縮した。残渣を酢酸エチルに溶かし、飽和ＮａＨＣＯ_３を注意深く加えた。有機層を単離し、Ｎａ_２ＳＯ_４で乾燥させ、濃縮した。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.44 (1 H, d, J=1.01 Hz), 8.63 - 8.86 (2 H, m), 8.51 (1 H, d, J=1.01 Hz), 7.38 (1 H, d, J=1.26 Hz), 7.17 (2 H, br. s.). MS (LC/MS) R.T. = 0.37; [M+H]⁺= 174.23。 Step C: 6- (pyrazin-2-yl) pyrimidin-4-amine

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 room temperature. The mixture was stirred at room temperature for 18 hours and concentrated. The residue was dissolved in ethyl acetate and saturated NaHCO ₃ was carefully added. The organic layer was isolated, dried over Na ₂ SO ₄ and concentrated.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.44 (1 H, d, J = 1.01 Hz), 8.63-8.86 (2 H, m), 8.51 (1 H, d, J = 1.01 Hz), 7.38 (1 H, d, J = 1.26 Hz), 7.17 (2 H, br. S.). MS (LC / MS) RT = 0.37; [M + H] ⁺ = 174.23.

ＤＭＦ中の６−(ピラジン−２−イル)ピリミジン−４−アミン(０.２ｇ, １.１５５mmol)の溶液に、ＮａＯＨ(０.１１５ml, ２.３１０mmol)、ＣＳ_２(０.１７４ml, ２.８９mmol)、ＮａＯＨ(０.１１５ml, ２.３１０mmol)およびＭｅＩ(０.１８１ml, ２.８９mmol)を、１５分間で加えた。１.５時間撹拌を続け、混合物を水に注いだ。橙色の固体を分離し、水で洗浄し、乾燥させ、明黄色の固体を得た(０.１４ｇ, ０.５０５mmol, ４４％)。
MS (LC/MS) R.T. = 3.19; [M+H]⁺= 278.19。 Step D: Dimethyl 6- (pyrazin-2-yl) pyrimidin-4-ylcarbonimidodithioate

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 ₂ (0.174 ml, 2.155). 89 mmol), NaOH (0.115 ml, 2.310 mmol) and MeI (0.181 ml, 2.89 mmol) were added over 15 minutes. Stirring was continued for 1.5 hours and the mixture was poured into water. The orange solid was separated, washed with water and dried to give a light yellow solid (0.14 g, 0.505 mmol, 44%).
MS (LC / MS) RT = 3.19; [M + H] ⁺ = 278.19.

Ｎ,Ｎ−ジメチルホルムアミド(１０ml)中の、６−(ピラジン−２−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(０.１３９ｇ, ０.５０１mmol)、Ｃｓ_２ＣＯ_３(０.４０８ｇ, １.２５３mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１１５ｇ, ０.５０１mmol)の混合物を、８０℃で３時間加熱した。混合物を濃縮し、シリカゲルで、酢酸エチル中０〜１０％ ９.５：０.５ メタノール：水酸化アンモニウムを用いて精製し、(Ｒ)−Ｎ−(６−(ピラジン−２−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを灰白色の固体として得た(０.０６６ｇ, ０.１９４mmol, ３９％)。
¹H NMR (400 MHz, MeOD) δ ppm 9.46 (1 H, d, J=1.51 Hz), 8.83 (1 H, d, J=1.26 Hz), 8.67 - 8.72 (1 H, m), 8.66 (1 H, d, J=2.52 Hz), 7.77 (1 H, br. s.), 4.05 (1 H, d, J=10.32 Hz), 3.75 (1 H, d, J=10.32 Hz), 3.18 - 3.26 (1 H, m), 3.04 - 3.15 (1 H, m), 2.87 - 3.00 (2 H, m), 2.70 - 2.85 (2 H, m), 2.04 - 2.19 (2 H, m), 1.54 - 1.83 (3 H, m). MS (LC/MS) R.T. = 0.97; [M+H]⁺= 338.29。 Step E: (R) -N- (6- (pyrazin-2-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

N, N- dimethylformamide in (10 ml), 6- (pyrazin-2-yl) pyrimidin-4-yl-carboxylic imide dithio dimethyl _{(0.139g, 0.501mmol), Cs 2} CO 3 (0.408g, 1.253 mmol) and (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (0.115 g, 0.501 mmol) were heated at 80 ° C. for 3 h. The mixture was concentrated and purified on silica gel using 0-10% in ethyl acetate 9.5: 0.5 methanol: ammonium hydroxide and (R) -N- (6- (pyrazin-2-yl) pyrimidine. -4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as an off-white solid (0.066 g, 0.194 mmol, 39%).
¹ H NMR (400 MHz, MeOD) δ ppm 9.46 (1 H, d, J = 1.51 Hz), 8.83 (1 H, d, J = 1.26 Hz), 8.67-8.72 (1 H, m), 8.66 (1 H, d, J = 2.52 Hz), 7.77 (1 H, br.s.), 4.05 (1 H, d, J = 10.32 Hz), 3.75 (1 H, d, J = 10.32 Hz), 3.18-3.26 (1 H, m), 3.04-3.15 (1 H, m), 2.87-3.00 (2 H, m), 2.70-2.85 (2 H, m), 2.04-2.19 (2 H, m), 1.54-1.83 (3 H, m). MS (LC / MS) RT = 0.97; [M + H] ⁺ = 338.29.

Example 335
(2R) -N- (6,7-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole] -2 '-Amine

メタノール(４.９ml)中の２,２−ジエトキシアセトイミド酸メチル(１.５０ｇ, ９.３２mmol)の溶液に、(３,４−ジメチルフェニル)メタンアミン(１.２０ｇ, ８.８８mmol)を環境温度で滴下した。反応フラスコを予め加熱した油浴に入れ、７０℃で１６時間撹拌した。この後、フラスコを油浴から取り出し、環境温度まで冷却した。揮発性成分を減圧下で除去し、粗製の物質を硫酸(１９.７ml)に環境温度で滴下した。反応混合物を７２時間撹拌し、フラスコを氷−水浴に入れ、水(５０ml)で希釈し、水酸化ナトリウム(１０Ｎ)でｐＨ＝１０までゆっくりと中和した。反応混合物が塩基性になったとき、橙色の沈殿物が形成した。この沈殿物を濾過し、水で洗浄し、乾燥させ、６,７−ジメチルイソキノリン−３−アミンおよび５,６−ジメチルイソキノリン−３−アミンの混合物(２：１)を得た(１.６８ｇ, ９.７５mmol, ＞１００％)。
MS (LC/MS) R.T. = 0.32; [M+H]⁺ = 173.13。 Step A: 6,7-dimethylisoquinolin-3-amine

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). Dropped at ambient temperature. The reaction flask was placed in a preheated oil bath and stirred at 70 ° C. for 16 hours. After this, the flask was removed from the oil bath and cooled to ambient temperature. Volatile components 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 hours, the flask was placed in an ice-water bath, diluted with water (50 ml) and slowly neutralized to pH = 10 with sodium hydroxide (10N). An orange precipitate formed when the reaction mixture became basic. The precipitate was filtered, washed with water and dried to give 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) RT = 0.32; [M + H] ⁺ = 173.13.

ジクロロメタン(１９ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１.３５ｇ, ５.８１mmol)の溶液に、環境温度で、６,７−ジメチルイソキノリン−３−アミンおよび５,６−ジメチルイソキノリン−３−アミンの混合物(２：１)(１.００ｇ, ５.８１mmol)を加えた。反応混合物を予め加熱した油浴中に置き、４０℃で１８時間撹拌し、冷却し、濃縮し、シリカゲルのクロマトグラフィーによって精製し(ヘキサン中１０〜３５％ 酢酸エチル)、３−イソチオシアナト−６,７−ジメチルイソキノリンおよび３−イソチオシアナト−５,６−ジメチルイソキノリンの混合物(２：１)を黄色の固体として得た(１８６mg, ０.８６９mmol, １５％)。
MS (LC/MS) R.T. = 2.06; [M+H]⁺ = 215.1。 Step B: 3-isothiocyanato-6,7-dimethylisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (1.35 g, 5.81 mmol) in dichloromethane (19 ml) at ambient temperature 6,7-dimethylisoquinolin-3-amine and A mixture of 5,6-dimethylisoquinolin-3-amine (2: 1) (1.00 g, 5.81 mmol) was added. The reaction mixture was placed in a preheated oil bath, stirred at 40 ° C. for 18 hours, cooled, concentrated, purified by chromatography on silica gel (10-35% ethyl acetate in hexane), 3-isothiocyanato-6, A mixture (2: 1) of 7-dimethylisoquinoline and 3-isothiocyanato-5,6-dimethylisoquinoline was obtained as a yellow solid (186 mg, 0.869 mmol, 15%).
MS (LC / MS) RT = 2.06; [M + H] ⁺ = 215.1.

Ｎ,Ｎ−ジメチルホルムアミド(１.４ml)中の(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(９９mg, ０.４３mmol)に、トリエチルアミン(０.１８１ml, １.３０mmol)と、３−イソチオシアナト−６,７−ジメチルイソキノリンおよび３−イソチオシアナト−５,６−ジメチルイソキノリンの混合物(２：１)(９３mg, ０.４３mmol)を加えた。懸濁液を、予め加熱した油浴中に置き、８０℃で２時間３０分撹拌した。Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.２７０ml, １.７４mmol)を加え、混合物を８０℃で１６時間撹拌した。混合物を濃縮し、シリカゲルのクロマトグラフィーによって精製し(クロロホルム中０〜４０％ ９：１ メタノール：水酸化アンモニウム)、次に逆相分取ＨＰＬＣによって精製した(０〜４０％ ＴＦＡ−メタノール−水)。生成物のフラクションを合わせ、真空で濃縮し、(２Ｒ)−Ｎ−(６,７−ジメチル−３−イソキノリニル)−４'Ｈ−スピロ[４−アザビシクロ[２.２.２]オクタン−２,５'−[１,３]オキサゾール]−２'−アミンおよび(２Ｒ)−Ｎ−(５,６−ジメチル−３−イソキノリニル)−４'Ｈ−スピロ[４−アザビシクロ[２.２.２]オクタン−２,５'−[１,３]オキサゾール]−２'−アミンの混合物(２：１)をトリフルオロ酢酸塩として得た。Chiralpak AD-H (4.6×250mm, 5μｍ)カラムを用いて、ＣＯ_２中４０％ メタノール(０.１％ ＤＥＡ)からなる移動相で、２１５nmで検出しながら２種の位置異性体を分離し、(２Ｒ)−Ｎ−(６,７−ジメチル−３−イソキノリニル)−４'Ｈ−スピロ[４−アザビシクロ[２.２.２]オクタン−２,５'−[１,３]オキサゾール]−２'−アミンを白色の固体として得た(５.０mg, ０.０１５mmol, 収率３％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.87 (1 H, s), 7.90 (1 H, s), 7.67 (1 H, s), 7.50 (1 H, s), 3.97 (1 H, d, J=10.0 Hz), 3.65 (1 H, d, J=10.0 Hz), 3.19 - 3.27 (1 H, m), 3.04 - 3.17 (1 H, m), 2.94 (2 H, d, J=7.5 Hz), 2.76 - 2.90 (2 H, m), 2.38 - 2.50 (6 H, m), 2.11 - 2.21 (2 H, m), 1.57 - 1.85 (3 H, m). MS (LC/MS) R.T. = 0.92; [M+H]⁺ = 337.22。 Step C: (2R) -N- (6,7-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole ] -2'-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (99 mg, 0.43 mmol) in N, N-dimethylformamide (1.4 ml) was added to triethylamine (0.181 ml, 1.30 mmol). And a mixture of 2-isothiocyanato-6,7-dimethylisoquinoline and 3-isothiocyanato-5,6-dimethylisoquinoline (2: 1) (93 mg, 0.43 mmol). The suspension was placed in a preheated oil bath and stirred at 80 ° C. for 2 hours 30 minutes. N, N′-diisopropylcarbodiimide (0.270 ml, 1.74 mmol) was added and the mixture was stirred at 80 ° C. for 16 hours. The mixture was concentrated and purified by chromatography on silica gel (0-40% 9: 1 methanol: ammonium hydroxide in chloroform) and then purified by reverse phase preparative HPLC (0-40% TFA-methanol-water). . The product fractions were combined and concentrated in vacuo to give (2R) -N- (6,7-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2, 5 '-[1,3] oxazole] -2'-amine and (2R) -N- (5,6-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] A mixture (2: 1) of octane-2,5 ′-[1,3] oxazole] -2′-amine was obtained as the trifluoroacetate salt. Using a Chiralpak AD-H (4.6 × 250 mm, 5 μm) column, a mobile phase consisting of 40% methanol in CO ₂ (0.1% DEA) was used to separate the two regioisomers while detecting at 215 nm, (2R) -N- (6,7-dimethyl-3-isoquinolinyl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole] -2 The '-amine was obtained as a white solid (5.0 mg, 0.015 mmol, 3% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.87 (1 H, s), 7.90 (1 H, s), 7.67 (1 H, s), 7.50 (1 H, s), 3.97 (1 H, d, J = 10.0 Hz), 3.65 (1 H, d, J = 10.0 Hz), 3.19-3.27 (1 H, m), 3.04-3.17 (1 H, m), 2.94 (2 H, d, J = 7.5 Hz ), 2.76-2.90 (2 H, m), 2.38-2.50 (6 H, m), 2.11-2.21 (2 H, m), 1.57-1.85 (3 H, m). MS (LC / MS) RT = 0.92; [M + H] ⁺ = 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 ] Oxazole] -2'-amine

オーブン乾燥した丸底フラスコに、ジ(１Ｈ−イミダゾール−１−イル)メタンイミン(１.４０ｇ, ８.６９mmol)、２−アミノ−５−メチルフェノール(７１３mg, ５.７９mmol)および無水ＴＨＦ(２０ml)を環境温度で入れた。得られた懸濁液をＮ_２(ｇ)下で２時間還流した。溶媒を真空で除去し、残渣をシリカゲルのクロマトグラフィーによって精製し(０〜３０％ ９：１ メタノール：水酸化アンモニウム−クロロホルム)、ベンゾ[ｄ]オキサゾール−２−アミンを灰色の固体として得た(７９２mg, ５.３５mmol, 収率９２％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 7.19 - 7.34 (1 H, m), 7.11 (1 H, s), 7.01 (1 H, d, J=7.8 Hz), 5.60 (2 H, br. s.), 2.43 (3 H, s). MS (LC/MS) R.T. = 0.89; [M+H]⁺ = 149.09。 Step A: 6-Methylbenzo [d] oxazol-2-amine

An oven-dried round bottom 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 ₂ (g) for 2 hours. The solvent was removed in vacuo and the residue was purified by chromatography on silica gel (0-30% 9: 1 methanol: ammonium hydroxide-chloroform) to give benzo [d] oxazol-2-amine as a gray solid ( 792 mg, 5.35 mmol, 92% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 7.19-7.34 (1 H, m), 7.11 (1 H, s), 7.01 (1 H, d, J = 7.8 Hz), 5.60 (2 H, br s.), 2.43 (3 H, s). MS (LC / MS) RT = 0.89; [M + H] ⁺ = 149.09.

ＤＭＦ(１.４ml)中の６−メチルベンゾ[ｄ]オキサゾール−２−アミン(２００mg, １.３５mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(１３５μｌ, ２.７０mmol)を加えた。混合物を室温で１０分間撹拌した後、二硫化炭素(２０３μｌ, ３.３７mmol)を加え、混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(１３５μｌ, ２.７０mmol)を加え、混合物を再度１０分間撹拌した。最後に、ヨードメタン(２０３μｌ, ３.２４mmol)を滴下した。添加完了後、混合物を１５分間撹拌し、この時点で多量の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、水で洗浄し、乾燥させ、６−メチルベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチルを褐色の固体として得た(２８９mg, １.１４mmol, 収率８５％)。
MS (LC/MS) R.T. = 1.88; [M+H]⁺ = 252.95。 Step B: Dimethyl 6-methylbenzo [d] oxazol-2-ylcarbonimidodithioate

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 μl, 2.70 mmol). After the mixture was stirred at room temperature for 10 minutes, carbon disulfide (203 μl, 3.37 mmol) was added and the mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (135 μl, 2.70 mmol) was added and the mixture was stirred again for 10 minutes. Finally, iodomethane (203 μl, 3.24 mmol) was added dropwise. After the addition was complete, the mixture was stirred for 15 minutes, at which point a large amount of precipitate formed. The mixture was poured into water and the solid was collected by filtration, washed with water and dried to give dimethyl 6-methylbenzo [d] oxazol-2-ylcarbonimidodithioate as a brown solid (289 mg, 1.14 mmol, Yield 85%).
MS (LC / MS) RT = 1.88; [M + H] ⁺ = 252.95.

１０mlのバイアルに、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール塩酸塩(３８.２mg, ０.１９８mmol)、ＤＭＦ(２ml)、Ｃｓ_２ＣＯ_３(１２９mg, ０.３９６mmol)および６−メチルベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチル(５０mg, ０.１９８mmol)を環境温度で入れた。得られた懸濁液を１時間撹拌した後、それをメタノールで希釈し、逆相分取ＨＰＬＣによって精製し(０〜１００％ ＴＦＡ−メタノール−水)、トリフルオロ酢酸塩として(２Ｒ)−Ｎ−(６−メチル−１,３−ベンゾオキサゾール−２−イル)−４'Ｈ−スピロ[４−アザビシクロ[２.２.２]オクタン−２,５'−[１,３]オキサゾール]−２'−アミンを黄褐色のゴム状物質として得た(５５.３mg, ０.１２３mmol, 収率６２％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.89 (br. s., 1 H), 9.15 (br. s., 1 H), 7.33 (d, J=8.03 Hz, 1 H), 7.31 (s, 1 H), 7.06 (d, J=8.78 Hz, 1 H), 4.00 (d, J=8.0 Hz, 1 H), 3.88 (d, J=10.29 Hz, 1 H), 3.64 - 3.80 (m, 2 H), 3.34 - 3.44 (m, 1 H), 3.20 - 3.34 (m, 3 H), 2.44 (br. s., 1 H), 2.40 (s, 3 H), 2.08 - 2.20 (m, 1 H), 1.80 - 2.03 (m, 3 H). MS (LC/MS) R.T. = 0.367, [M+H]⁺ = 313.2。 Step C: (2R) -N- (6-Methyl-1,3-benzoxazol-2-yl) -4′H-spiro [4-azabicyclo [2.2.2] octane-2,5 ′-[ 1,3] oxazole] -2′-amine

Vial 10 ml, (S) -3- (aminomethyl) quinuclidine-3-ol hydrochloride (38.2mg, 0.198mmol), DMF ( 2ml), Cs 2 CO 3 (129mg, 0.396mmol) and 6 -Dimethyl benzo [d] oxazol-2-ylcarbonimidodithioate (50 mg, 0.198 mmol) was charged at ambient temperature. The resulting suspension was stirred for 1 hour before it was diluted with methanol and purified by reverse phase preparative HPLC (0-100% TFA-methanol-water) as (2R) -N as the trifluoroacetate salt. -(6-Methyl-1,3-benzoxazol-2-yl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3] oxazole] -2 The '-amine was obtained as a tan gum (55.3 mg, 0.123 mmol, 62% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.89 (br. S., 1 H), 9.15 (br. S., 1 H), 7.33 (d, J = 8.03 Hz, 1 H), 7.31 (s, 1 H), 7.06 (d, J = 8.78 Hz, 1 H), 4.00 (d, J = 8.0 Hz, 1 H), 3.88 (d, J = 10.29 Hz, 1 H), 3.64-3.80 ( m, 2 H), 3.34-3.44 (m, 1 H), 3.20-3.34 (m, 3 H), 2.44 (br. s., 1 H), 2.40 (s, 3 H), 2.08-2.20 (m , 1 H), 1.80-2.03 (m, 3 H). MS (LC / MS) RT = 0.367, [M + H] ⁺ = 313.2.

Example 337
(2R) -N- (6-Methoxy-1,3-benzoxazol-2-yl) -4'H-spiro [4-azabicyclo [2.2.2] octane-2,5 '-[1,3 ] Oxazole] -2'-amine

オーブン乾燥した丸底フラスコに、ジ(１Ｈ−イミダゾール−１−イル)メタンイミン(５００mg, ３.１０mmol)、２−アミノ−５−メトキシフェノール−ＨＣｌ(３６５mg, ２.０７mmol)、トリエチルアミン(２８８μｌ, ２.０７mmol)および無水ＴＨＦ(２０ml)を環境温度で入れた。得られた懸濁液をＮ_２(ｇ)下で２０時間還流し、ＬＣ／ＭＳに基づいて変換の完了を見た。溶媒を真空で除去し、残渣をシリカゲルのクロマトグラフィーによって精製し(ヘキサン中３０〜８０％ 酢酸エチル)、６−メトキシベンゾ[ｄ]オキサゾール−２−アミンを褐色の固体として得た(３０７mg, １.８７mmol, 収率９０％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 7.28 (1 H, d, J=9.5 Hz), 6.92 (1 H, d, J=2.4 Hz), 6.80 (1 H, dd, J=8.5, 2.1 Hz), 5.28 (2 H, br. s.). MS (LC/MS) R.T. = 1.51; [M+H]⁺ = 165.00。 Step A: 6-Methoxybenzo [d] oxazol-2-amine

In an oven-dried round bottom flask, di (1H-imidazol-1-yl) methanimine (500 mg, 3.10 mmol), 2-amino-5-methoxyphenol-HCl (365 mg, 2.07 mmol), triethylamine (288 μl, 2 0.07 mmol) and anhydrous THF (20 ml) were charged at ambient temperature. The resulting suspension was refluxed under N ₂ (g) for 20 hours and saw 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 give 6-methoxybenzo [d] oxazol-2-amine as a brown solid (307 mg, 1 .87 mmol, 90% yield).
¹ H NMR (500 MHz, chloroform-d) δ ppm 7.28 (1 H, d, J = 9.5 Hz), 6.92 (1 H, d, J = 2.4 Hz), 6.80 (1 H, dd, J = 8.5, 2.1 Hz), 5.28 (2 H, br. S.). MS (LC / MS) RT = 1.51; [M + H] ⁺ = 165.00.

ＤＭＦ(２.０ml)中の、６−メトキシベンゾ[ｄ]オキサゾール−２−アミン(２３８mg, １.４５mmol)の懸濁液に、２０.０Ｍ 水酸化ナトリウム(１４５μｌ, ２.９０mmol)を加えた。混合物を室温で１０分間撹拌した後、二硫化炭素(２１９μｌ, ３.６２mmol)を加え、混合物を１０分間撹拌した。さらに２０.０Ｍ 水酸化ナトリウム(１４５μｌ, ２.９０mmol)を加え、混合物を再度１０分間撹拌した。最後にヨードメタン(２１８μｌ, ３.４８mmol)を滴下した。混合物をさらに１５分間撹拌し、この時点で多量の沈殿物が形成した。混合物を水に注ぎ、固体を濾過によって集め、水で洗浄し、乾燥させ、６−メトキシベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチルを得た。この物質を特性決定せずに用いた。 Step B: Dimethyl 6-methoxybenzo [d] oxazol-2-ylcarbonimidodithioate

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 μl, 2.90 mmol). . After the mixture was stirred at room temperature for 10 minutes, carbon disulfide (219 μl, 3.62 mmol) was added and the mixture was stirred for 10 minutes. Further 20.0M sodium hydroxide (145 μl, 2.90 mmol) was added and the mixture was stirred again for 10 minutes. Finally iodomethane (218 μl, 3.48 mmol) was added dropwise. The mixture was stirred for an additional 15 minutes, at which point a large amount of precipitate formed. The mixture was poured into water and the solid was collected by filtration, washed with water and dried to give dimethyl 6-methoxybenzo [d] oxazol-2-ylcarbonimidodithioate. This material was used without characterization.

１０mlのバイアルに、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール塩酸塩(２４.５mg, ０.１２７mmol)、ＤＭＦ(２ml)、Ｃｓ_２ＣＯ_３(８３mg, ０.２５mmol)および６−メトキシベンゾ[ｄ]オキサゾール−２−イルカルボンイミドジチオ酸ジメチル(３４mg, ０.１３mmol)を環境温度で入れた。得られた懸濁液を環境温度で１時間撹拌し、メタノールで希釈し、逆相分取ＨＰＬＣによって精製し(０〜１００％ ＴＦＡ−メタノール−水)、トリフルオロ酢酸塩として(Ｒ)−Ｎ−(６−メトキシベンゾ[ｄ]オキサゾール−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを黄褐色のゴム状物質として得た(７.２mg, ０.０１４mmol, 収率１１％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.95 (br. s., 1 H), 9.10 (br. s., 1 H), 7.34 (d, J=8.78 Hz, 1 H), 7.17 (d, J=2.51 Hz, 1 H), 6.84 (dd, J=8.53, 2.51 Hz, 1 H), 3.99 (d, J=10.54 Hz, 1 H), 3.87 (d, J=10.54 Hz, 1 H), 3.79 (s, 3 H), 3.63 - 3.78 (m, 2 H), 3.34 - 3.44 (m, 1 H), 3.20 - 3.32 (m, 3 H), 2.43 (m, 1 H), 2.09 - 2.19 (m, 1 H), 1.91 - 2.02 (m, 1 H), 1.80 - 1.91 (m, 2 H). MS (LC/MS) R.T. = 0.867, [M+H]⁺ = 329.28。 Step C: (2R) -N- (6-Methoxy-1,3-benzoxazol-2-yl) -4′H-spiro [4-azabicyclo [2.2.2] octane-2,5 ′-[ 1,3] oxazole] -2′-amine

Vial 10 ml, (S) -3- (aminomethyl) quinuclidine-3-ol hydrochloride (24.5mg, 0.127mmol), DMF ( 2ml), Cs 2 CO 3 (83mg, 0.25mmol) and 6 -Dimethyl methoxybenzo [d] oxazol-2-ylcarbonimidodithioate (34 mg, 0.13 mmol) was charged at ambient temperature. The resulting suspension was stirred at ambient temperature for 1 hour, diluted with methanol, purified by reverse phase preparative HPLC (0-100% TFA-methanol-water) and (R) -N as the trifluoroacetate salt. -(6-Methoxybenzo [d] oxazol-2-yl) -4H-1'-azaspiro [oxazol-5,3'-bicyclo [2.2.2] octane] -2-amine as a tan gum Obtained as material (7.2 mg, 0.014 mmol, 11% yield).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.95 (br. S., 1 H), 9.10 (br. S., 1 H), 7.34 (d, J = 8.78 Hz, 1 H), 7.17 (d, J = 2.51 Hz, 1 H), 6.84 (dd, J = 8.53, 2.51 Hz, 1 H), 3.99 (d, J = 10.54 Hz, 1 H), 3.87 (d, J = 10.54 Hz, 1 H), 3.79 (s, 3 H), 3.63-3.78 (m, 2 H), 3.34-3.44 (m, 1 H), 3.20-3.32 (m, 3 H), 2.43 (m, 1 H), 2.09 -2.19 (m, 1 H), 1.91-2.02 (m, 1 H), 1.80-1.91 (m, 2 H). MS (LC / MS) RT = 0.867, [M + H] ⁺ = 329.28.

Examples 338A and 338B
(R) -N- (5-chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine (A) and ( R) -N- (7-Chloroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine (B)

窒素を吹き付けた密封した試験管に、メタノール(６ml)および(３−クロロフェニル)メタンアミン(１.４４４ml, １１.８２mmol)中の２,２−ジエトキシアセトイミド酸メチル(２ｇ, １２.４１mmol)を入れた。試験管に蓋をして、７０℃で加熱した。１８時間後、反応物を室温まで冷却し、反応物を濃縮し、Ｎ−(３−クロロベンジル)−２,２−ジエトキシアセトイミドアミドを黄色の油状物として得た(２.３ｇ, ８.５mmol, ７１％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.14 - 7.50 (m, 4 H) 4.80 (br. s., 1 H) 4.29 (br. s., 2 H) 4.05 (q, J=7.11 Hz, 1 H) 3.40 - 3.73 (m, 4 H) 2.01 (s, 1 H) 1.05 - 1.29 (m, 6 H). LC/MS RT = 1.12分, [M+H] = 271.09。 Step A: N- (3-chlorobenzyl) -2,2-diethoxyacetimidoamide

To a sealed tube, sparged with nitrogen, methyl 2,2-diethoxyacetimidate (2 g, 12.41 mmol) in methanol (6 ml) and (3-chlorophenyl) methanamine (1.444 ml, 11.82 mmol) is added. I put it in. The test tube was capped and heated at 70 ° C. After 18 hours, the reaction was cooled to room temperature and the reaction was concentrated to give N- (3-chlorobenzyl) -2,2-diethoxyacetimidoamide as a yellow oil (2.3 g, 8 .5 mmol, 71%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 7.14-7.50 (m, 4 H) 4.80 (br. S., 1 H) 4.29 (br. S., 2 H) 4.05 (q, J = 7.11 Hz, 1 H) 3.40-3.73 (m, 4 H) 2.01 (s, 1 H) 1.05-1.29 (m, 6 H). LC / MS RT = 1.12 min, [M + H] = 271.09.

フラスコに、Ｎ−(３−クロロベンジル)−２,２−ジエトキシアセトイミドアミド(１.９ｇ, ７.０２mmol)および硫酸(１４ml, ２６３mmol)を入れた。これを室温で２４時間撹拌した。１０Ｎ ＮａＯＨを用いてｐＨ ９まで反応物を中和した。得られた沈殿物をジクロロメタンに溶解し、水層をジクロロメタンで洗浄した。有機層を合わせ、濃縮し、２：１比の７−クロロイソキノリン−３−アミン：５−クロロイソキノリン−３−アミンで、上記の位置異性体(１.１ｇ, ６.１mmol, ８８％)を得た。この位置異性体混合物をさらに分離することなく用いた。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.89 (s, 1 H) 8.81 (s, 2 H) 7.90 (d, J=2.26 Hz, 2 H) 7.82 (d, J=8.28 Hz, 1 H) 7.65 (dd, J=7.28, 1.00 Hz, 1 H) 7.58 (d, J=9.03 Hz, 2 H) 7.44 (dd, J=9.03, 2.26 Hz, 2 H) 7.07 - 7.19 (m, 1 H) 6.83 (s, 1 H) 6.64 (s, 2 H) 6.28 (s, 2 H) 6.05 (s, 4 H). LC/MS R.T. = 0.955分, [M+H] = 181.03。 Step B: 5- and 7-chloro-isoquinolin-3-amine

A flask was charged with N- (3-chlorobenzyl) -2,2-diethoxyacetimidoamide (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 neutralized to pH 9 with 10N NaOH. The obtained precipitate was dissolved in dichloromethane, and the aqueous layer was washed with dichloromethane. The organic layers were combined and concentrated to give the above regioisomer (1.1 g, 6.1 mmol, 88%) with a 2: 1 ratio of 7-chloroisoquinolin-3-amine: 5-chloroisoquinolin-3-amine. Obtained. This regioisomer mixture was used without further separation.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.89 (s, 1 H) 8.81 (s, 2 H) 7.90 (d, J = 2.26 Hz, 2 H) 7.82 (d, J = 8.28 Hz, 1 H) 7.65 (dd, J = 7.28, 1.00 Hz, 1 H) 7.58 (d, J = 9.03 Hz, 2 H) 7.44 (dd, J = 9.03, 2.26 Hz, 2 H) 7.07-7.19 (m, 1 H ) 6.83 (s, 1 H) 6.64 (s, 2 H) 6.28 (s, 2 H) 6.05 (s, 4 H). LC / MS RT = 0.955 min, [M + H] = 181.03.

バイアルに、ジクロロメタン(１０ml)中の７−クロロイソキノリン−３−アミン／５−クロロイソキノリン−３−アミン(５５０mg, ３.０８mmol)および１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(８５８mg, ３.７０mmol)を入れた。反応物を室温で撹拌した。１８時間後、反応物をシリカゲルのカートリッジで精製し、ヘキサン中５０％ 酢酸エチルで溶出し、７−クロロ−３−イソチオシアナトイソキノリンおよび５−クロロ−３−イソチオシアナトイソキノリンの混合物を得た(４００mg, １.８mmol, ５８％)。これをすぐに次の反応に用いた。
LC/MS RT = 3.808分, [M+H] = 221.0。 Step C: 5- and 7-chloro-3-isothiocyanato-isoquinoline

A vial is charged with 7-chloroisoquinolin-3-amine / 5-chloroisoquinolin-3-amine (550 mg, 3.08 mmol) and 1,1′-thiocarbonyldipyridin-2 (1H) -one in dichloromethane (10 ml). (858 mg, 3.70 mmol) was added. The reaction was stirred at room temperature. After 18 hours, the reaction was purified on a silica gel cartridge eluting with 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%). This was used immediately in the next reaction.
LC / MS RT = 3.808 min, [M + H] = 221.0.

バイアルに、ＤＭＦ(８ml)中の７−クロロ−３−イソチオシアナトイソキノリン／５−クロロ−３−イソチオシアナトイソキノリン(４００mg, １.８１３mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール(３４０mg, ２.１７５mmol)を入れた。これに炭酸セシウム(１４１７mg, ４.３５mmol)を加えた。反応物を５０℃で１時間撹拌し、Ｎ,Ｎ'−ジイソプロピルカルボジイミド(０.８４７ml, ５.４４mmol)を反応物に加え、反応物を一夜撹拌した。１８時間後、反応物を水およびクロロホルムに注いだ。有機物を集めて濃縮し、Biotageで精製し、クロロホルム中５〜４０％ (１０％ ＮＨ_４ＯＨ／メタノール)で溶出した。生成物の位置異性体を集め、分取ＳＦＣによって分離し、(Ｒ)−Ｎ−(５−クロロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミン(１２.５mg)を明黄色の固体として単離した。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.14 (s, 1 H) 8.67 (br. s., 1 H) 8.03 (d, J=8.28 Hz, 1 H) 7.82 (d, J=7.28 Hz, 1 H) 7.41 (t, J=8.03 Hz, 1 H) 3.90 (br. s., 1 H) 3.63 (br. s., 1 H) 2.91 - 3.16 (m, 2 H) 2.65 - 2.92 (m, 4 H) 1.85 - 2.19 (m, 2 H) 1.16 - 1.81 (m, 4 H). LC/MS RT=1.578分, [M+H]=343.08。 Step D: (R) -N- (5-Chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine and ( R) -N- (7-Chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

A vial is charged with 7-chloro-3-isothiocyanatoisoquinoline / 5-chloro-3-isothiocyanatoisoquinoline (400 mg, 1.813 mmol) and (S) -3- (aminomethyl) quinuclidine-in DMF (8 ml). 3-ol (340 mg, 2.175 mmol) was added. To this was added cesium carbonate (1417 mg, 4.35 mmol). The reaction was stirred at 50 ° C. for 1 hour, N, N′-diisopropylcarbodiimide (0.847 ml, 5.44 mmol) was added to the reaction and the reaction was stirred overnight. After 18 hours, the reaction was poured into water and chloroform. The organics were collected, concentrated, and purified by Biotage, eluting with 5-40% in chloroform (10% _NH 4 OH / methanol). The product regioisomers were collected and separated by preparative SFC, and (R) -N- (5-chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2 2.2.2] Octane] -2-amine (12.5 mg) was isolated as a light yellow solid.
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.14 (s, 1 H) 8.67 (br. S., 1 H) 8.03 (d, J = 8.28 Hz, 1 H) 7.82 (d, J = 7.28 Hz, 1 H) 7.41 (t, J = 8.03 Hz, 1 H) 3.90 (br. S., 1 H) 3.63 (br. S., 1 H) 2.91-3.16 (m, 2 H) 2.65-2.92 ( m, 4 H) 1.85-2.19 (m, 2 H) 1.16-1.81 (m, 4 H). LC / MS RT = 1.578 min, [M + H] = 343.08.

And (R) -N- (7-chloroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine (54. 9 mg) was isolated as a light yellow solid.
1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.06 (s, 1 H) 8.43-8.93 (m, 1 H) 8.13 (br. S., 1 H) 7.82 (br. S., 1 H) 7.61 (d, J = 8.28 Hz, 1 H) 3.89 (br. s., 1 H) 3.61 (br. s., 1 H) 3.31 (br. s., 1 H) 2.90-3.18 (m, 2 H) 2.62-2.95 (m, 4 H) 1.82-2.13 (m, 2 H) 1.34-1.80 (m, 3 H). LC / MS RT = 0.898 min, [M + H] = 343.32.

Example 339
(R) -N- (6-chlorothiazolo [4,5-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

アセトン(２０ml)中の３,５−ジクロロピリジン−２−アミン(３.８７ｇ, ２３.７３mmol)に、ベンゾイル イソチオシアネート(３.２ml, ２３.７３mmol)を加えた。混合物を環境温度で１時間撹拌し、得られた沈殿物を濾過によって集めた(３.８９ｇ)。濾液を反応フラスコに戻し、１時間還流し、この時点で、反応物が乾固した。固体をアセトンに懸濁し、濾過によって集め、２.２４ｇのＮ−(３,５−ジクロロピリジン−２−イルカルバモチオイル)ベンズアミドを得た(５.９３ｇ, １８.１８mmol, 収率７７％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 12.78 (br. s., 1 H) 9.25 (br. s., 1 H) 8.46 (br. s., 1 H) 7.94 (d, J=7.63 Hz, 2 H) 7.88 (d, J=2.44 Hz, 1 H) 7.69 (t, J=7.48 Hz, 1 H) 7.58 (t, J=7.78 Hz, 2 H)。 Step A: N- (3,5-dichloropyridin-2-ylcarbamothioyl) benzamide

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 hour and the resulting precipitate was collected by filtration (3.89 g). The filtrate was returned to the reaction flask and refluxed for 1 hour, at which point the reaction was dry. The solid was 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). .
¹ H NMR (500 MHz, chloroform-d) δ ppm 12.78 (br. S., 1 H) 9.25 (br. S., 1 H) 8.46 (br. S., 1 H) 7.94 (d, J = 7.63 Hz, 2 H) 7.88 (d, J = 2.44 Hz, 1 H) 7.69 (t, J = 7.48 Hz, 1 H) 7.58 (t, J = 7.78 Hz, 2 H).

ＮＭＰ(４０ml)中のＮ−(３,５−ジクロロピリジン−２−イルカルバモチオイル)ベンズアミド(５.２ｇ, １６mmol)に、ナトリウム メトキシド(１.７３ｇ, ３２.０mmol)を加えた。混合物を１２０℃の油浴中で４時間加熱し、環境温度まで冷却し、水に注いだ。固体を濾過によって集め、シリカゲルのクロマトグラフィーによって精製し(０〜５％ (９：１ ＭｅＯＨ：ＮＨ_４ＯＨ)／クロロホルム)、Ｎ−(６−クロロチアゾロ[４,５−ｂ]ピリジン−２−イル)ベンズアミドを得た(３６４mg, １.２５６mmol, 収率７.８５％)。
¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.16 (br. s., 1 H) 8.48 - 8.58 (m, 2 H) 8.13 - 8.23 (m, 2 H) 7.61 - 7.69 (m, 1 H) 7.56 (t, J=7.53 Hz, 2 H). MS (LC/MS) R.T. = 2.8; [M+H]⁺ = 290.1。 Step B: N- (6-chlorothiazolo [4,5-b] pyridin-2-yl) benzamide

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 an oil bath at 120 ° C. for 4 hours, cooled to ambient temperature and poured into water. The solid was collected by filtration and purified by chromatography on silica gel (0-5% (9: 1 MeOH: NH ₄ OH) / chloroform) and N- (6-chlorothiazolo [4,5-b] pyridin-2-yl. Benzamide was obtained (364 mg, 1.256 mmol, yield 7.85%).
¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 13.16 (br. S., 1 H) 8.48-8.58 (m, 2 H) 8.13-8.23 (m, 2 H) 7.61-7.69 (m, 1 H ) 7.56 (t, J = 7.53 Hz, 2 H). MS (LC / MS) RT = 2.8; [M + H] ⁺ = 290.1.

Ｈ_２ＳＯ_４(４ml, ７５mmol)中のＮ−(６−クロロチアゾロ[４,５−ｂ]ピリジン−２−イル)ベンズアミド(３６４mg, １.２５６mmol)を、１２０℃で３０分間加熱し、環境温度まで冷却し、１０Ｎ ＮａＯＨで塩基性にし、得られた固体を濾過によって集め、６−クロロチアゾロ[４,５−ｂ]ピリジン−２−アミンを得た(１４８mg, ０.７９７mmol, 収率６３.５％)。
¹H NMR (500 MHz, DMSO-d₆) δ ppm 8.19 - 8.24 (m, 2 H) 8.09 (br. s., 2 H)。 Step C: 6-chlorothiazolo [4,5-b] pyridin-2-amine

N- (6-chlorothiazolo [4,5-b] pyridin-2-yl) benzamide (364 mg, 1.256 mmol) in H ₂ SO ₄ (4 ml, 75 mmol) was heated at 120 ° C. for 30 minutes to ambient temperature. And cooled to 10 N NaOH, the resulting solid was collected by filtration to give 6-chlorothiazolo [4,5-b] pyridin-2-amine (148 mg, 0.797 mmol, 63.5 yield). %).
¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 8.19-8.24 (m, 2 H) 8.09 (br. S., 2 H).

６−クロロチアゾロ[４,５−ｂ]ピリジン−２−アミン(１４８mg, ０.７９７mmol)をＤＭＦ(１ml)に懸濁し、１６Ｎ ＮａＯＨ(０.１００ml, １.５９５mmol)を加えた。混合物を環境温度で１０分間撹拌し、この時点で、ＣＳ_２(０.１２０ml, １.９９３mmol)を加えた。混合物を環境温度でさらに１０分間撹拌し、１６Ｎ ＮａＯＨ(０.１００ml, １.５９５mmol)を加えた。混合物を環境温度で１０分間撹拌し、この時点で、ヨードメタン(０.１２０ml, １.９１３mmol)を加えた。１５分間後、混合物を水で希釈し、得られた固体を濾過によって集めた。濾液をクロロホルムで３回抽出し、硫酸ナトリウムで乾燥させた。粗製の混合物をシリカゲルのクロマトグラフィーによって精製し(２〜５０％ ＥｔＯＡｃ／ＣＨＣｌ_３)、６−クロロチアゾロ[４,５−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルを得た(５０mg, ０.１７３mmol, 収率２１.６４％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.57 (s, 1 H) 8.09 (s, 1 H) 2.66 (s, 6 H)。 Step D: Dimethyl 6-chlorothiazolo [4,5-b] pyridin-2-ylcarbonimidodithioate

6-chlorothiazolo [4,5-b] pyridin-2-amine (148 mg, 0.797 mmol) was suspended in DMF (1 ml) and 16N NaOH (0.100 ml, 1.595 mmol) was added. The mixture was stirred at ambient temperature for 10 minutes, at which point CS ₂ (0.120 ml, 1.993 mmol) was added. The mixture was stirred at ambient temperature for an additional 10 minutes and 16N NaOH (0.100 ml, 1.595 mmol) was added. The mixture was stirred at ambient temperature for 10 minutes, at which point iodomethane (0.120 ml, 1.913 mmol) was added. After 15 minutes, the mixture was diluted with water and the resulting solid was collected by filtration. The filtrate was extracted 3 times with chloroform and dried over sodium sulfate. The crude mixture was purified by chromatography on silica gel (2-50% EtOAc / CHCl ₃ ) to give dimethyl 6-chlorothiazolo [4,5-b] pyridin-2-ylcarbonimidodithioate (50 mg, 0.0. 173 mmol, yield 21.64%).
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.57 (s, 1 H) 8.09 (s, 1 H) 2.66 (s, 6 H).

６−クロロチアゾロ[４,５−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(５０mg, ０.１７mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(４０mg, ０.１７mmol)および炭酸セシウム(１１２mg, ０.３５mmol)をＤＭＦ(１ml)に懸濁し、開放フラスコ中、１００℃の油浴で加熱した。２.５時間後、混合物を環境温度まで冷却し、水に注いだ。混合物をクロロホルム(４×)で抽出し、硫酸ナトリウムで乾燥させ、濾過し、濃縮して残渣を得た。粗製の残渣を、シリカゲルのクロマトグラフィーによって、５〜４０％ (９：１ ＭｅＯＨ／ＮＨ_４ＯＨ)／ＣＨＣｌ_３で精製し、(Ｒ)−Ｎ−(６−クロロチアゾロ[４,５−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(２０mg, ０.０５６mmol, 収率３２.５％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.31 (d, J=2.51 Hz, 1 H) 7.93 (d, J=2.26 Hz, 1 H) 3.99 (d, J=10.04 Hz, 1 H) 3.67 (d, J=10.04 Hz, 1 H) 3.20 - 3.39 (m, 2 H) 2.86 - 3.05 (m, 3 H) 2.67 - 2.87 (m, 2 H) 2.03 - 2.19 (m, 2 H) 1.74 (ddd, J=13.87, 9.35, 4.64 Hz, 1 H) 1.44 - 1.65 (m, 2 H). MS (LC/MS) R.T. = 1.49; [M+H]⁺ = 350.2。 Step E: (R) -N- (6-chlorothiazolo [4,5-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 6-chlorothiazolo [4,5-b] pyridin-2-ylcarbonimidodithioate (50 mg, 0.17 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (40 mg, 0 .17 mmol) and cesium carbonate (112 mg, 0.35 mmol) were suspended in DMF (1 ml) and heated in a 100 ° C. oil bath in an open flask. After 2.5 hours, the mixture was cooled to ambient temperature and poured into water. The mixture was extracted with chloroform (4x), dried over sodium sulfate, filtered and concentrated to give a residue. The crude residue by chromatography on silica gel, 5-40% was purified by _{(9 1 MeOH / NH 4 OH} ) / CHCl 3, (R) -N- (6- chlorothiazolo [4,5-b] pyridine -2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (20 mg, 0.056 mmol, yield). 32.5%).
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.31 (d, J = 2.51 Hz, 1 H) 7.93 (d, J = 2.26 Hz, 1 H) 3.99 (d, J = 10.04 Hz, 1 H) 3.67 (d, J = 10.04 Hz, 1 H) 3.20-3.39 (m, 2 H) 2.86-3.05 (m, 3 H) 2.67-2.87 (m, 2 H) 2.03-2.19 (m, 2 H) 1.74 (ddd , J = 13.87, 9.35, 4.64 Hz, 1 H) 1.44-1.65 (m, 2 H). MS (LC / MS) RT = 1.49; [M + H] ⁺ = 350.2.

Example 340
(R) -N- (5-Fluorothiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

機械的に撹拌しながら、酢酸(８９ml)中のロダン酸カリウム(２７.７ｇ, ２８５mmol)の懸濁液を含む３ツ首フラスコに、６−フルオロピリジン−３−アミン(４ｇ, ３５.７mmol)を０℃で加えた。酢酸(２９.７ml)中の臭素(５.７０ml, １１１mmol)を入れた滴下漏斗をフラスコに取り付けた。臭素溶液を３０分かけて加えると、溶液は粘性の黄色の混合物となった。臭素の添加が完了した後、反応混合物を環境温度まで温め、１６時間撹拌した。水(３０ml)を加え、混合物を８５℃で２０分間加熱した後、固体を濾過し、水で、そしてメタノールで洗浄し、５−フルオロチアゾロ[５,４−ｂ]ピリジン−２−アミンを黄色の固体として得た(４.１８ｇ, ２４.７１mmol, 収率６９.２％)。 Step A: 5-Fluorothiazolo [5,4-b] pyridin-2-amine

To a three-necked flask containing a suspension of potassium rhodanoate (27.7 g, 285 mmol) in acetic acid (89 ml) with mechanical stirring, was added 6-fluoropyridin-3-amine (4 g, 35.7 mmol). Was added at 0 ° C. A dropping funnel containing bromine (5.70 ml, 111 mmol) in acetic acid (29.7 ml) was attached to the flask. The bromine solution was added over 30 minutes and the solution became a viscous yellow mixture. After complete addition of bromine, the reaction mixture was warmed to ambient temperature and stirred for 16 hours. After adding water (30 ml) and heating the mixture at 85 ° C. for 20 minutes, the solid is filtered, washed with water and with methanol to give 5-fluorothiazolo [5,4-b] pyridin-2-amine. Obtained as a yellow solid (4.18 g, 24.71 mmol, 69.2% yield).

５−フルオロチアゾロ[５,４−ｂ]ピリジン−２−アミン(２.０ｇ, １１.８mmol)を、実施例３３９の工程Ｄの方法に従って反応させ、５−フルオロチアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチルを黄色の固体として得た(２.１７ｇ, 収率６７％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.14 (dd, J=8.55, 7.02 Hz, 1 H) 6.98 (dd, J=8.70, 1.98 Hz, 1 H) 2.63 (s, 6 H)。 Step B: Dimethyl 5-fluorothiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate

5-Fluorothiazolo [5,4-b] pyridin-2-amine (2.0 g, 11.8 mmol) is reacted according to the method of Example 339, Step D to give 5-fluorothiazolo [5,4- b] Dimethyl pyridine-2-ylcarbonimidodithioate was obtained as a yellow solid (2.17 g, 67% yield).
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.14 (dd, J = 8.55, 7.02 Hz, 1 H) 6.98 (dd, J = 8.70, 1.98 Hz, 1 H) 2.63 (s, 6 H).

５−フルオロチアゾロ[５,４−ｂ]ピリジン−２−イルカルボンイミドジチオ酸ジメチル(１００mg, ０.３７mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(８４mg, ０.３７mmol)および炭酸セシウム(２３８mg, ０.７３mmol)を、ＤＭＦ(２ml)に懸濁し、開放フラスコ中で、１００℃の油浴で加熱した。２.５時間後、混合物を環境温度まで冷却し、水に注いだ。混合物をクロロホルム(４×)で抽出し、硫酸ナトリウムで乾燥させ、濾過し、濃縮した。粗製の残渣を、シリカゲルのクロマトグラフィーによって、５〜４０％ (９：１ ＭｅＯＨ／ＮＨ_４ＯＨ)／ＣＨＣｌ_３で精製し、((Ｒ)−Ｎ−(５−フルオロチアゾロ[５,４−ｂ]ピリジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(８５mg, 収率６６％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.17 (br. s., 1 H) 7.89 (dd, J=8.66, 6.90 Hz, 1 H) 6.91 (dd, J=8.53, 2.01 Hz, 1 H) 4.04 (d, J=9.54 Hz, 1 H) 3.73 (d, J=9.54 Hz, 1 H) 3.43 (dd, J=14.93, 1.63 Hz, 1 H) 3.08 (dd, J=15.06, 1.76 Hz, 1 H) 2.84 - 3.05 (m, 4 H) 2.16 - 2.28 (m, 2 H) 1.75 - 1.87 (m, 1 H) 1.53 - 1.70 (m, 2 H). MS (LC/MS) R.T. = 1.37; [M+H]⁺ = 334.2。 Step C: (R) -N- (5-Fluorothiazolo [5,4-b] pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

Dimethyl 5-fluorothiazolo [5,4-b] pyridin-2-ylcarbonimidodithioate (100 mg, 0.37 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (84 mg , 0.37 mmol) and cesium carbonate (238 mg, 0.73 mmol) were suspended in DMF (2 ml) and heated in a 100 ° C. oil bath in an open flask. After 2.5 hours, the mixture was cooled to ambient temperature and poured into water. The mixture was extracted with chloroform (4x), dried over sodium sulfate, filtered and concentrated. The crude residue by chromatography on silica gel, 5-40% was purified by _{(9 1 MeOH / NH 4 OH} ) / CHCl 3, ((R) -N- (5- fluoro thiazolo [5,4- b] Pyridin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (85 mg, 66 yield). %).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.17 (br. S., 1 H) 7.89 (dd, J = 8.66, 6.90 Hz, 1 H) 6.91 (dd, J = 8.53, 2.01 Hz, 1 H ) 4.04 (d, J = 9.54 Hz, 1 H) 3.73 (d, J = 9.54 Hz, 1 H) 3.43 (dd, J = 14.93, 1.63 Hz, 1 H) 3.08 (dd, J = 15.06, 1.76 Hz, 1 H) 2.84-3.05 (m, 4 H) 2.16-2.28 (m, 2 H) 1.75-1.87 (m, 1 H) 1.53-1.70 (m, 2 H). MS (LC / MS) RT = 1.37; [M + H] ⁺ = 334.2.

Example 341
(R) -N- (6-Methylthiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

クロロホルム(１００ml)中の５−メチルピラジン−２−アミン(１.０９ｇ, １０mmol)の懸濁液に、ピリジン(０.８５ml, １０.５mmol)を加えた。滴下漏斗を取り付けたホイルで包んだフラスコ中で混合物を撹拌し、クロロホルム(１０ml)中の臭素(０.５４ml, １０.５mmol)の溶液を１０分かけて滴下した。添加が完了した後、混合物をさらに２０分間反応させ、１０mlの水を含む分液漏斗に注いだ。相を分離し、有機物を再度水で洗浄し、硫酸ナトリウムで乾燥させ、濾過し、真空で濃縮した。得られた赤色の油状物を、シリカゲルのクロマトグラフィーによって、１２〜１００％ ＥｔＯＡｃ／ヘキサンで精製した。主要なＵＶ活性ピークを集め、３−ブロモ−５−メチルピラジン−２−アミンをクリーム色の固体として得た(１.０６ｇ, ５.６４mmol, 収率５６.４％)。
¹H NMR (500 MHz, DMSO-d6) δ ppm 7.85 (s, 1 H) 6.42 (br. s., 2 H) 2.25 (s, 3 H). MS (LC/MS) R.T. = 0.93; [M+H]⁺ = 189.9。 Step A: 3-Bromo-5-methylpyrazin-2-amine

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 a dropping funnel and a solution of bromine (0.54 ml, 10.5 mmol) in chloroform (10 ml) was added dropwise over 10 minutes. After the addition was complete, the mixture was allowed to react for an additional 20 minutes and poured into a separatory funnel containing 10 ml of water. The phases were separated and the organics were washed again with water, dried over sodium sulfate, filtered and concentrated in vacuo. The resulting red oil was purified by chromatography on silica gel with 12-100% EtOAc / hexanes. The major UV active peak was collected to give 3-bromo-5-methylpyrazin-2-amine as a cream solid (1.06 g, 5.64 mmol, 56.4% yield).
¹ H NMR (500 MHz, DMSO-d6) δ ppm 7.85 (s, 1 H) 6.42 (br. S., 2 H) 2.25 (s, 3 H). MS (LC / MS) RT = 0.93; (M + H] ⁺ = 189.9.

３−ブロモ−５−メチルピラジン−２−アミン(８５０mg, ４.５２mmol)に、エトキシカルボニル イソチオシアネート(０.５１ml, ４.５２mmol)を、次にトルエン(１ml)を加えた。混合物を、予め加熱した１００℃の油浴中に１分以内で置き、全ての固体を溶解させた。１５分後、全量を整粒された固体の塊とした。さらに１５分間後、メタノール(約５ml)を加え、混合物を還流して固体を破壊した。５分後に混合物を室温まで冷却し、固体を濾過によって集め、メタノールで洗浄し、６−メチルチアゾロ[５,４−ｂ]ピラジン−２−イルカルバミン酸エチルを得た(６８０mg, ２.８５mmol, 収率６３.１％)。
¹H NMR (400 MHz, DMSO-d6) δ ppm 12.48 (br. s., 1 H) 8.48 (s, 1 H) 4.30 (q, J=7.03 Hz, 2 H) 2.56 - 2.65 (m, 3 H) 1.31 (t, J=7.15 Hz, 3 H). MS (LC/MS) R.T. = 2.10 ; [M+H]⁺ = 239.17。 Step B: Ethyl 6-methylthiazolo [5,4-b] pyrazin-2-ylcarbamate

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 ° C. oil bath within 1 minute to dissolve all solids. After 15 minutes, the entire amount was made into a sized solid lump. After an additional 15 minutes, methanol (ca. 5 ml) was added and the mixture was refluxed to destroy the solid. After 5 minutes the mixture was cooled to room temperature and the solid was collected by filtration and washed with methanol to give ethyl 6-methylthiazolo [5,4-b] pyrazin-2-ylcarbamate (680 mg, 2.85 mmol, yield). (Rate 63.1%).
¹ H NMR (400 MHz, DMSO-d6) δ ppm 12.48 (br. S., 1 H) 8.48 (s, 1 H) 4.30 (q, J = 7.03 Hz, 2 H) 2.56-2.65 (m, 3 H ) 1.31 (t, J = 7.15 Hz, 3 H). MS (LC / MS) RT = 2.10; [M + H] ⁺ = 239.17.

６−メチルチアゾロ[５,４−ｂ]ピラジン−２−イルカルバミン酸エチル(６６０mg, ２.７７mmol)を、１Ｎ ＮａＯＨ(１５ml, １５.００mmol)に懸濁し、１００℃の油浴中で４時間加熱し、環境温度まで冷却した。１Ｎ ＨＣｌで混合物を酸性にして、得られた沈殿物を濾過して取り、エーテルで洗浄し、６−メチルチアゾロ[５,４−ｂ]ピラジン−２−アミンを黄色の粉末として得た(２３３mg, １.４０２mmol, 収率５０.６％)。
¹H NMR (500 MHz, DMSO-d6) δ ppm 8.26 (s, 2 H) 8.12 (s, 1 H) 2.43 (s, 3 H). MS (LC/MS) R.T. = 0.66; [M+H]⁺ = 167.0。 Step C: 6-Methylthiazolo [5,4-b] pyrazin-2-amine

Ethyl 6-methylthiazolo [5,4-b] pyrazin-2-ylcarbamate (660 mg, 2.77 mmol) is suspended in 1N NaOH (15 ml, 15.00 mmol) and heated in a 100 ° C. oil bath for 4 hours. And cooled to ambient temperature. The mixture was acidified with 1N HCl and the resulting precipitate was filtered off and washed with ether to give 6-methylthiazolo [5,4-b] pyrazin-2-amine as a yellow powder (233 mg, 1.402 mmol, 50.6% yield).
¹ H NMR (500 MHz, DMSO-d6) δ ppm 8.26 (s, 2 H) 8.12 (s, 1 H) 2.43 (s, 3 H). MS (LC / MS) RT = 0.66; [M + H] ⁺ = 167.0.

６−メチルチアゾロ[５,４−ｂ]ピラジン−２−アミン(０.２２ｇ, １.３２mmol)を、実施例３３９の工程Ｄの方法に従って反応させ、６−メチルチアゾロ[５,４−ｂ]ピラジン−２−イルカルボンイミドジチオ酸ジメチルを黄色の固体として得た(２６５mg, 収率５９％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.45 (s, 1 H) 2.66 - 2.68 (m, 9 H)。 Step D: Dimethyl 6-methylthiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate

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 give 6-methylthiazolo [5,4-b] pyrazine- Dimethyl 2-ylcarbonimidodithioate was obtained as a yellow solid (265 mg, 59% yield).
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.45 (s, 1 H) 2.66-2.68 (m, 9 H).

６−メチルチアゾロ[５,４−ｂ]ピラジン−２−イルカルボンイミドジチオ酸ジメチル(１５０mg, ０.５６mmol)、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１４０mg, ０.６１mmol)および炭酸セシウム(４５０mg, １.３９mmol)をＤＭＦ(３ml)に懸濁し、開放フラスコ中、１００℃の油浴で加熱した。２.５時間後、混合物を環境温度まで冷却し、水に注いだ。混合物をクロロホルム(４×)で抽出し、硫酸ナトリウムで乾燥させ、濾過し、濃縮して残渣を得た。粗製の残渣を、シリカゲルのクロマトグラフィーによって、５〜４０％ (９：１ ＭｅＯＨ／ＮＨ_４ＯＨ)／ＣＨＣｌ_３で精製し、(Ｒ)−Ｎ−(６−メチルチアゾロ[５,４−ｂ]ピラジン−２−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを白色の粉末として得た(５１mg, 収率２７％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.45 (br. s., 1 H) 8.26 (s, 1 H) 4.02 (d, J=9.79 Hz, 1 H) 3.69 (d, J=9.79 Hz, 1 H) 3.39 (dd, J=14.93, 1.63 Hz, 1 H) 2.72 - 3.06 (m, 5 H) 2.60 (s, 3 H) 2.11 - 2.22 (m, 2 H) 1.70 - 1.82 (m, 1 H) 1.49 - 1.64 (m, 2 H). MS (LC/MS) R.T. = 1.15; [M+H]⁺ = 331.2。 Step E: (R) -N- (6-Methylthiazolo [5,4-b] pyrazin-2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane ] -2-Amine

Dimethyl 6-methylthiazolo [5,4-b] pyrazin-2-ylcarbonimidodithioate (150 mg, 0.56 mmol), (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (140 mg, 0 .61 mmol) and cesium carbonate (450 mg, 1.39 mmol) were suspended in DMF (3 ml) and heated in a 100 ° C. oil bath in an open flask. After 2.5 hours, the mixture was cooled to ambient temperature and poured into water. The mixture was extracted with chloroform (4x), dried over sodium sulfate, filtered and concentrated to give a residue. The crude residue by chromatography on silica gel, 5-40% was purified by _{(9 1 MeOH / NH 4 OH} ) / CHCl 3, (R) -N- (6- methylthiazolo [5,4-b] pyrazine -2-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained as a white powder (51 mg, 27% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.45 (br. S., 1 H) 8.26 (s, 1 H) 4.02 (d, J = 9.79 Hz, 1 H) 3.69 (d, J = 9.79 Hz , 1 H) 3.39 (dd, J = 14.93, 1.63 Hz, 1 H) 2.72-3.06 (m, 5 H) 2.60 (s, 3 H) 2.11-2.22 (m, 2 H) 1.70-1.82 (m, 1 H) 1.49-1.64 (m, 2 H). MS (LC / MS) RT = 1.15; [M + H] ⁺ = 331.2.

Example 342
(R) -N- (6,7-dimethoxyisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

２,２−ジエトキシアセトイミド酸メチル(２９４mg, １.８２４mmol)、(３,４−ジメトキシフェニル)メタンアミン(３１２mg, １.８６６mmol)およびＭｅＯＨ(１.５ml)の混合物を、撹拌しながら、油浴中、７０℃で２時間加熱し、室温まで冷却した。揮発性成分を真空で除去した。残渣をさらに精製することなく用いた。
LCMS RT 0.96分, MH⁺= 297.2, ¹H NMR (400 MHz, クロロホルム-d) δ ppm 6.79 - 6.93 (3 H, m), 4.94 (1 H, s), 4.41 (2 H, s), 3.84 - 3.90 (6 H, m), 3.52 - 3.70 (4 H, m), 1.19 - 1.28 (6 H, m)。 Step A: N- (3,4-dimethoxybenzyl) -2,2-diethoxyacetimidoamide

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 stirred with oil. Heat in a bath at 70 ° C. for 2 hours and cool to room temperature. Volatile components were removed in vacuo. The residue was used without further purification.
LCMS RT 0.96 min, MH ⁺ = 297.2, ¹ H NMR (400 MHz, chloroform-d) δ ppm 6.79-6.93 (3 H, m), 4.94 (1 H, s), 4.41 (2 H, s), 3.84 -3.90 (6 H, m), 3.52-3.70 (4 H, m), 1.19-1.28 (6 H, m).

Ｎ−(３,４−ジメトキシベンジル)−２,２−ジエトキシアセトイミドアミド(５４１mg, １.８２５mmol)に、硫酸(０.９５ml, １.８２５mmol)を加えた。冷却後、反応物を環境温度で一夜静置した。それを氷に滴下し、得られた溶液を濃ＮａＯＨで中和し、水相を酢酸エチルで２回抽出した。合わせた酢酸エチルフラクションを硫酸マグネシウムで乾燥させた。乾燥剤を濾過して除き、溶媒を蒸発させた。残渣をカラムクロマトグラフィーによって５％ メタノール／酢酸エチルで精製し、主な成分を集めた。１０５.７mgの褐色の固体を得た(２８％)。
LCMS RT 0.72分, MH⁺ = 205.1; 1H NMR (500 MHz, クロロホルム-d) δ ppm 8.59 (1 H, s), 7.03 (1 H, s), 6.80 (1 H, s), 6.70 (1 H, s), 4.00 (3 H, s), 3.98 (3 H, s)。 Step B: 6,7-Dimethoxyisoquinolin-3-amine

To N- (3,4-dimethoxybenzyl) -2,2-diethoxyacetimidamide (541 mg, 1.825 mmol) was added sulfuric acid (0.95 ml, 1.825 mmol). After cooling, the reaction was left overnight at ambient temperature. It was added dropwise to ice, the resulting solution was neutralized with concentrated NaOH, and the aqueous phase was extracted twice with ethyl acetate. The combined ethyl acetate fractions were dried over magnesium sulfate. The desiccant was filtered off and the solvent was evaporated. The residue was purified by column chromatography with 5% methanol / ethyl acetate and the main components were collected. 105.7 mg of a brown solid was obtained (28%).
LCMS RT 0.72 min, MH ⁺ = 205.1; 1H NMR (500 MHz, chloroform-d) δ ppm 8.59 (1 H, s), 7.03 (1 H, s), 6.80 (1 H, s), 6.70 (1 H , s), 4.00 (3 H, s), 3.98 (3 H, s).

ジクロロメタン(３ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１２４.６mg, ０.５３６mmol)の溶液に、撹拌しながら、ジクロロメタン(５ml)中の６,７−ジメトキシイソキノリン−３−アミン(１０５mg, ０.５１４mmol)の懸濁液を加えた。反応混合物を直接Biotageカラムにアプライし、１５〜２５％ 酢酸エチル／ヘキサンで、次に１００％ 酢酸エチルで精製し、３−イソチオシアナト−６,７−ジメトキシイソキノリンを白色の固体として得た。収量６９.４mg(５５％)。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.81 (1 H, s), 7.31 (1 H, s), 7.16 (1 H, s), 6.98 (1 H, s), 3.99 (6 H, s)。 Step C: 3-isothiocyanato-6,7-dimethoxyisoquinoline

To a solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (124.6 mg, 0.536 mmol) in dichloromethane (3 ml) with stirring, 6,7-dimethoxy in dichloromethane (5 ml). A suspension of isoquinolin-3-amine (105 mg, 0.514 mmol) was added. The reaction mixture was applied directly to a Biotage column and purified with 15-25% ethyl acetate / hexane and then with 100% ethyl acetate to give 3-isothiocyanato-6,7-dimethoxyisoquinoline as a white solid. Yield 69.4 mg (55%).
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.81 (1 H, s), 7.31 (1 H, s), 7.16 (1 H, s), 6.98 (1 H, s), 3.99 (6 H, s).

ＤＭＦ(５ml)中の塩化 (Ｓ)−３−(アンモニオメチル)−３−ヒドロキシ−１−アゾニアビシクロ[２.２.２]オクタン(７３mg, ０.３１９mmol)および炭酸セシウム(２２１mg, ０.６７８mmol)の懸濁液に、撹拌しながら、ＤＭＦ(１.５ml)中の３−イソチオシアナト−６,７−ジメトキシイソキノリン(６９mg, ０.２８０mmol)の溶液を加え、反応混合物を室温で３日間撹拌した。この溶液に、ＤＭＦ(０.４ml)中のＮ,Ｎ'−メタンジイリデンジプロパン−２−アミン(６９mg, ０.５４７mmol)の溶液を加え、反応物を室温で１４日間静置した。さらに約０.４mlのアセトニトリル中の６９mgのジ−イソプロピルカルボジイミドを加え、反応物を１日以上静置し、それを真空で蒸発させ、残渣について分取ＨＰＬＣを行い、(Ｒ)−Ｎ−(６,７−ジメトキシイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを２０.９mg(８％)得た。
LCMS RT 0.98分, MH⁺ = 403.2. ¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.73 (1 H, s), 7.28 (1 H, s), 7.09 (1 H, s), 6.96 (1 H, s), 4.01 (3 H, s), 4.00 (3 H, s), 3.96 (1 H, d, J=8.5 Hz), 3.63 (1 H, d, J=9.2 Hz), 3.39 (1 H, d, J=15.0 Hz), 2.79 - 3.10 (5 H, m), 2.24 (1 H, br. s.), 2.15 (1 H, br. s.), 1.75 (1 H, dddd, J=13.9, 9.3, 4.7, 4.4 Hz), 1.59 - 1.68 (1 H, m), 1.48 - 1.59 (1 H, m)。 Step D: (R) -N- (6,7-dimethoxyisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

(S) -3- (ammoniomethyl) -3-hydroxy-1-azoniabicyclo [2.2.2] octane (73 mg, 0.319 mmol) and cesium carbonate (221 mg, 0.678 mmol) in DMF (5 ml) ) With stirring, a solution of 3-isothiocyanato-6,7-dimethoxyisoquinoline (69 mg, 0.280 mmol) in DMF (1.5 ml) was added and the reaction mixture was stirred at room temperature for 3 days. . To this solution was added a solution of N, N′-methanediylidenedipropan-2-amine (69 mg, 0.547 mmol) in DMF (0.4 ml) and the reaction was left at room temperature for 14 days. An additional 69 mg of di-isopropylcarbodiimide in about 0.4 ml of acetonitrile was added, the reaction was allowed to stand for more than 1 day, it was evaporated in vacuo, the residue was subjected to preparative HPLC, and (R) -N- ( 6,7-Dimethoxyisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained in 20.9 mg (8%). .
LCMS RT 0.98 min, MH ⁺ = 403.2. ¹ H NMR (500 MHz, chloroform-d) δ ppm 8.73 (1 H, s), 7.28 (1 H, s), 7.09 (1 H, s), 6.96 (1 H, s), 4.01 (3 H, s), 4.00 (3 H, s), 3.96 (1 H, d, J = 8.5 Hz), 3.63 (1 H, d, J = 9.2 Hz), 3.39 (1 H, d, J = 15.0 Hz), 2.79-3.10 (5 H, m), 2.24 (1 H, br. S.), 2.15 (1 H, br. S.), 1.75 (1 H, dddd, J = 13.9, 9.3, 4.7, 4.4 Hz), 1.59-1.68 (1 H, m), 1.48-1.59 (1 H, m).

Example 343
(R) -N- (5,6,7,8-tetrahydroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2- Amine

イソキノリン−３−アミン(２３９mg, １.６６mmol)、酸化白金(IV)(２８mg, ０.１２３mmol)およびＴＦＡ(６ml)の混合物を、Parr社製装置中で３時間水素化した。反応混合物を酢酸エチルを用いて濾過した。濾液を真空で蒸発させ、残渣を１０％ 水性炭酸ナトリウムと酢酸エチルの層間に分配した。層を分離し、再度水相を酢酸エチルで洗浄し、合わせた有機層を塩水で洗浄し、硫酸マグネシウムで乾燥させた。乾燥剤を濾過して除き、溶媒を蒸発させた。この物質をカラムクロマトグラフィーによって酢酸エチルで精製し、１３９.８mg(５７％)の５,６,７,８−テトラヒドロイソキノリン−３−アミンを黄白色の固体として得た。
LCMS RT 0.75分, MH⁺ = 149.1. ¹H NMR (400 MHz, クロロホルム-d) δ ppm 7.78 (1 H, s), 6.26 (1 H, s), 4.29 (2 H, br. s.), 2.64 (4 H, ddd, J=10.6, 5.8, 5.5 Hz), 1.64 - 1.87 (4 H, m)。 Step A: 5,6,7,8-tetrahydroisoquinolin-3-amine

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 a Parr apparatus for 3 hours. The reaction mixture was filtered using 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 and the aqueous phase was washed again with ethyl acetate and the combined organic layers were washed with brine and dried over magnesium sulfate. The desiccant was filtered off and the solvent was evaporated. This material was purified by column chromatography with ethyl acetate to give 139.8 mg (57%) of 5,6,7,8-tetrahydroisoquinolin-3-amine as a pale yellow solid.
LCMS RT 0.75 min, MH ⁺ = 149.1. ¹ H NMR (400 MHz, chloroform-d) δ ppm 7.78 (1 H, s), 6.26 (1 H, s), 4.29 (2 H, br. S.), 2.64 (4 H, ddd, J = 10.6, 5.8, 5.5 Hz), 1.64-1.87 (4 H, m).

ジクロロメタン(５ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(２２２mg, ０.９５６mmol)の溶液に、撹拌しながら、ジクロロメタン(１.５ml)中の５,６,７,８−テトラヒドロイソキノリン−３−アミン(１３９mg, ０.９３８mmol)の溶液を加え、得られた溶液を室温で一夜撹拌した。反応混合物を直接Biotageカラムにアプライし、２０〜２５％ 酢酸エチル／ヘキサンで精製し、主要なピークを集めた。１４１.８mg(７９％)の３−イソチオシアナト−５,６,７,８−テトラヒドロイソキノリンを得た。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.10 (1 H, s), 6.84 (1 H, s), 2.74 (4 H, t, J=6.3 Hz), 1.81 (4 H, dd, J=4.0, 2.7 Hz)。 Step B: 3-isothiocyanato-5,6,7,8-tetrahydroisoquinoline

A solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (222 mg, 0.956 mmol) in dichloromethane (5 ml) was stirred with 5,6,7 in dichloromethane (1.5 ml). , 8-tetrahydroisoquinolin-3-amine (139 mg, 0.938 mmol) was added and the resulting solution was stirred at room temperature overnight. The reaction mixture was applied directly to a Biotage column and purified with 20-25% ethyl acetate / hexane to collect the main peak. 141.8 mg (79%) of 3-isothiocyanato-5,6,7,8-tetrahydroisoquinoline were obtained.
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.10 (1 H, s), 6.84 (1 H, s), 2.74 (4 H, t, J = 6.3 Hz), 1.81 (4 H, dd, J = 4.0, 2.7 Hz).

ＤＭＦ(１０ml)中の、塩化 (Ｓ)−３−(アンモニオメチル)−３−ヒドロキシ−１−アゾニアビシクロ[２.２.２]オクタン(１８３mg, ０.７９９mmol)および炭酸セシウム(５５０mg, １.６８８mmol)の懸濁液に、撹拌しながら、ＤＭＦ(２ml)中の３−イソチオシアナト−５,６,７,８−テトラヒドロイソキノリン(１４１mg, ０.７４１mmol)の溶液を加え、反応混合物を室温で１０日間撹拌した。溶媒を真空で除去し、残渣をメタノールに溶かし、シリカゲルのカラムで１％ ＮＨ_４ＯＨ／９％ ＭｅＯＨ／９０％ ＣＨＣｌ_３で、次に分取ＨＰＬＣで精製した。単離した生成物を水性炭酸ナトリウムと酢酸エチルの層間に分配した。水相を酢酸エチルで再度洗浄し、合わせた有機相を塩水で洗浄し、硫酸マグネシウムで乾燥させ、濾過し、溶媒を蒸発させた。残渣を、シリカゲルのカラムで、１％ ＮＨ_４ＯＨ／９％ ＭｅＯＨ／９０％ ＣＨＣｌ_３で再度精製し、(Ｓ)−１−((３−ヒドロキシキヌクリジン−３−イル)メチル)−３−(５,６,７,８−テトラヒドロイソキノリン−３−イル)チオウレアを９８mg(３５％)得た。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 12.02 (1 H, br s), 8.23 (1 H, br s), 7.88 (1 H, s), 6.41 (1 H, s), 4.08 (1 H, dd), 3.89 (1 H, dd), 3.46 (2 H, s), 3.05-2.80 (5 H, m), 2.69 (4 H, dt), 2.10 (1 H, br s), 1.93 (1 H, br s), 1.85 (1 H, m), 1.77 (4 H, m), 1.66 (1 H, m), 1.45 (1 H, m); LCMS RT 1.11分, MH⁺ = 447.1。 Step C: (S) -1-((3-hydroxyquinuclidin-3-yl) methyl) -3- (5,6,7,8-tetrahydroisoquinolin-3-yl) thiourea

(S) -3- (Ammoniomethyl) -3-hydroxy-1-azoniabicyclo [2.2.2] octane (183 mg, 0.799 mmol) and cesium carbonate (550 mg, 1.99 ml) in DMF (10 ml). To a suspension of 688 mmol) a solution of 3-isothiocyanato-5,6,7,8-tetrahydroisoquinoline (141 mg, 0.741 mmol) in DMF (2 ml) is added and the reaction mixture is stirred at room temperature for 10 minutes. Stir for days. The solvent was removed in vacuo and the residue was dissolved in methanol and purified on a silica gel column with 1% NH ₄ OH / 9% MeOH / 90% CHCl ₃ and then by 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 was evaporated. The residue was purified again on a silica gel column with 1% NH ₄ OH / 9% MeOH / 90% CHCl ₃ to give (S) -1-((3-hydroxyquinuclidin-3-yl) methyl) -3 98 mg (35%) of-(5,6,7,8-tetrahydroisoquinolin-3-yl) thiourea were obtained.
¹ H NMR (500 MHz, chloroform-d) δ ppm 12.02 (1 H, br s), 8.23 (1 H, br s), 7.88 (1 H, s), 6.41 (1 H, s), 4.08 (1 H, dd), 3.89 (1 H, dd), 3.46 (2 H, s), 3.05-2.80 (5 H, m), 2.69 (4 H, dt), 2.10 (1 H, br s), 1.93 ( 1 H, br s), 1.85 (1 H, m), 1.77 (4 H, m), 1.66 (1 H, m), 1.45 (1 H, m); LCMS RT 1.11 min, MH ⁺ = 447.1.

ＤＭＦ(２ml)中の(Ｓ)−１−((３−ヒドロキシキヌクリジン−３−イル)メチル)−３−(５,６,７,８−テトラヒドロイソキノリン−３−イル)チオウレア(９８mg, ０.２８３mmol)の溶液に、ＤＭＦ(０.４ml)中のＮ,Ｎ'−メタンジイリデンジプロパン−２−アミン(４１mg, ０.３２５mmol)の溶液を加え、得られた溶液を室温で７日間静置した。さらに約０.４mlのアセトニトリル中の５３mgのジ−イソプロピルカルボジイミドを加え、反応物を１日以上静置した。反応混合物を真空で蒸発させ、分取ＨＰＬＣを行い、次に１％ ＮＨ_４ＯＨ／９％ ＭｅＯＨ／９０％ ＣＨＣｌ_３でシリカゲルのクロマトグラフィーを行い、(Ｒ)−Ｎ−(５,６,７,８−テトラヒドロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを６.４mg(７％)得た。
LCMS RT 0.80分, MH⁺ =313.2; ¹H NMR (500 MHz, クロロホルム-d) δ ppm 7.91 (1 H, s), 6.75 - 6.95 (1 H, m), 3.91 (1 H, d, J=9.5 Hz), 3.60 (1 H, d, J=9.5 Hz), 3.38 (1 H, s), 2.93 - 3.13 (4 H, m), 2.86 - 2.93 (2 H, m), 2.69 (4 H, td, J=11.4, 6.1 Hz), 2.21 - 2.32 (1 H, m), 2.16 (1 H, br. s.), 1.72 - 1.85 (5 H, m), 1.64 (1 H, dd, J=7.2, 4.4 Hz), 1.56 (1 H, dt, J=7.0, 2.6 Hz)。 Step D: (R) -N- (5,6,7,8-tetrahydroisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -1-((3-hydroxyquinuclidin-3-yl) methyl) -3- (5,6,7,8-tetrahydroisoquinolin-3-yl) thiourea (98 mg, DMF (2 ml)) To a solution of 0.283 mmol) 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 added at room temperature for 7 Let stand for days. A further 53 mg of di-isopropylcarbodiimide in about 0.4 ml of acetonitrile was added and the reaction was allowed to stand for more than 1 day. The reaction mixture was evaporated in vacuo, preparative HPLC was performed, followed by chromatography on silica gel with 1% NH ₄ OH / 9% MeOH / 90% CHCl ₃ and (R) -N- (5,6,7 , 8-Tetrahydroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained in an amount of 6.4 mg (7%).
LCMS RT 0.80 min, MH ⁺ = 313.2; ¹ H NMR (500 MHz, chloroform-d) δ ppm 7.91 (1 H, s), 6.75-6.95 (1 H, m), 3.91 (1 H, d, J = 9.5 Hz), 3.60 (1 H, d, J = 9.5 Hz), 3.38 (1 H, s), 2.93-3.13 (4 H, m), 2.86-2.93 (2 H, m), 2.69 (4 H, td, J = 11.4, 6.1 Hz), 2.21-2.32 (1 H, m), 2.16 (1 H, br. s.), 1.72-1.85 (5 H, m), 1.64 (1 H, dd, J = 7.2, 4.4 Hz), 1.56 (1 H, dt, J = 7.0, 2.6 Hz).

Example 344
(R) -N- (6-Chloro-7-methoxyisoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(４−クロロ−３−メトキシフェニル)メタンアミン(３９２mg, ２.２８４mmol)(塩酸塩として)、ヒューニッヒ塩基(０.５ml, ２.８６mmol)およびＭｅＯＨ(２ml)の混合物に、ＭｅＯＨ(１ml)中の２,２−ジエトキシアセトイミド酸メチル(２９５mg, １.８３mmol)の溶液を加え、得られた混合物を、撹拌しながら、油浴中、７０℃で２時間加熱し、３日かけて室温まで冷却した。溶媒を真空で除去し、残渣をさらに精製することなく用いた。 Step A: N- (4-Chloro-3-methoxybenzyl) -2,2-diethoxyacetimidoamide

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) in MeOH (1 ml). A solution of methyl 2,2-diethoxyacetimidate (295 mg, 1.83 mmol) was added and the resulting mixture was heated in an oil bath at 70 ° C. for 2 hours with stirring and allowed to reach room temperature over 3 days. Cooled down. The solvent was removed in vacuo and the residue was used without further purification.

冷却(０℃)した硫酸(０.０７１ml, １.３３０mmol)に、ジクロロメタン(５ml)中のＮ−(４−クロロ−３−メトキシベンジル)−２,２−ジエトキシアセトイミドアミド(４００mg, １.３３０mmol)の懸濁液を徐々に加え、得られた混合物を一夜かけて室温まで温めた。反応混合物を砕いた氷にゆっくりと加え、濃ＮａＯＨ溶液を添加することによって混合物を強塩基性にし、有機成分を酢酸エチルで抽出した。有機フラクションを硫酸マグネシウムで乾燥させ、濾過し、溶媒を蒸発させた。残渣についてシリカゲルのクロマトグラフィーを酢酸エチルで行い、１１０.８mg(４０％)の６−クロロ−７−メトキシイソキノリン−３−アミンを得た。
LCMS RT 0.95分, MH⁺ =209.0, 211.0; ¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.66 (1 H, s), 7.78 (1 H, s), 6.83 (1 H, s), 6.63 (1 H, s), 4.36 - 4.58 (2 H, m), 3.99 (3 H, s)。 Step B: 6-Chloro-7-methoxyisoquinolin-3-amine

To cooled (0 ° C.) sulfuric acid (0.071 ml, 1.330 mmol) was added N- (4-chloro-3-methoxybenzyl) -2,2-diethoxyacetimidoamide (400 mg, 1) in dichloromethane (5 ml). .330 mmol) suspension was added slowly and the resulting mixture was allowed to warm to room temperature overnight. The reaction mixture was slowly added to crushed ice, the mixture was made strongly basic by adding concentrated NaOH solution, and the organic components were extracted with ethyl acetate. The organic fraction was dried over magnesium sulfate, filtered and the solvent was evaporated. The residue was chromatographed on silica gel with ethyl acetate to give 110.8 mg (40%) of 6-chloro-7-methoxyisoquinolin-3-amine.
LCMS RT 0.95 min, MH ⁺ = 209.0, 211.0; ¹ H NMR (500 MHz, chloroform-d) δ ppm 8.66 (1 H, s), 7.78 (1 H, s), 6.83 (1 H, s), 6.63 (1 H, s), 4.36-4.58 (2 H, m), 3.99 (3 H, s).

ジクロロメタン(５ml)中の１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１４２mg, ０.６１１mmol)の溶液に、撹拌しながら、ジクロロメタン(５.００ml)中の６−クロロ−７−メトキシイソキノリン−３−アミン(１１０mg, ０.５２７mmol)の懸濁液を加え、反応混合物を室温で一夜撹拌した。反応混合物を直接シリカゲルカラムにアプライし、１０％ 酢酸エチル／ヘキサンで精製し、１０６mg(８０％)の６−クロロ−３−イソチオシアナト−７−メトキシイソキノリンを得た。
¹H NMR (500 MHz, クロロホルム-d) δ ppm 8.90 (1 H, s), 7.99 (1 H, s), 7.36 (1 H, s), 7.08 (1 H, s), 4.05 (3 H, s)。 Step C: 6-Chloro-3-isothiocyanato-7-methoxyisoquinoline

A solution of 1,1′-thiocarbonyldipyridin-2 (1H) -one (142 mg, 0.611 mmol) in dichloromethane (5 ml) is stirred with 6-chloro-7 in dichloromethane (5.00 ml). A suspension of -methoxyisoquinolin-3-amine (110 mg, 0.527 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was applied directly to a silica gel column and purified with 10% ethyl acetate / hexane to give 106 mg (80%) of 6-chloro-3-isothiocyanato-7-methoxyisoquinoline.
¹ H NMR (500 MHz, chloroform-d) δ ppm 8.90 (1 H, s), 7.99 (1 H, s), 7.36 (1 H, s), 7.08 (1 H, s), 4.05 (3 H, s).

ＤＭＦ(３ml)中の塩化 (Ｓ)−３−(アンモニオメチル)−３−ヒドロキシ−１−アゾニアビシクロ[２.２.２]オクタン(１１１mg, ０.４８４mmol)の懸濁液に、撹拌しながら、ヒューニッヒ塩基(０.５ml, ２.８６mmol)を加え、得られた混合物を室温で３５分間撹拌した。ＤＭＦ(３ml)中の６−クロロ−３−イソチオシアナト−７−メトキシイソキノリン(１０６mg, ０.４２３mmol)の懸濁液を加え、反応混合物を室温で一夜撹拌した。溶媒を真空で蒸発させ、残渣を、シリカゲルのカラムで、１％ ＮＨ_４ＯＨ／９％ ＭｅＯＨ／９０％ ＣＨＣｌ_３で精製し、白色の固体として、１４０.３mg(８２％)の(Ｓ)−１−(６−クロロ−７−メトキシイソキノリン−３−イル)−３−((３−ヒドロキシキヌクリジン−３−イル)メチル)チオウレアを得た。
LCMS RT 1.28分, MH⁺ = 407.1, 409.0, ¹H NMR (500 MHz, クロロホルム-d) δ ppm 11.85 (1 H, t, J=5.2 Hz), 9.15 (1 H, br. s.), 8.63 (1 H, s), 7.76 (1 H, s), 6.92 (1 H, s), 6.89 (1 H, s), 4.14 (1 H, dd, J=13.9, 5.6 Hz), 3.99 (2 H, s), 3.90 (1 H, dd, J=14.0, 5.2 Hz), 3.46 (3 H, s), 2.89 - 3.01 (2 H, m), 2.75 - 2.88 (3 H, m), 2.03 - 2.15 (1 H, m), 1.89 - 1.96 (1 H, m), 1.77 - 1.88 (1 H, m), 1.63 (1 H, ddd, J=13.4, 6.1, 3.4 Hz), 1.32 - 1.45 (1 H, m)。 Step D: (S) -1- (6-Chloro-7-methoxyisoquinolin-3-yl) -3-((3-hydroxyquinuclidin-3-yl) methyl) thiourea

To a suspension of (S) -3- (ammoniomethyl) -3-hydroxy-1-azoniabicyclo [2.2.2] octane (111 mg, 0.484 mmol) in DMF (3 ml) with stirring. Hunig's base (0.5 ml, 2.86 mmol) was added and the resulting mixture was stirred at room temperature for 35 minutes. A suspension of 6-chloro-3-isothiocyanato-7-methoxyisoquinoline (106 mg, 0.423 mmol) in DMF (3 ml) was added and the reaction mixture was stirred at room temperature overnight. The solvent was evaporated in vacuo and the residue was purified on a silica gel column with 1% NH ₄ OH / 9% MeOH / 90% CHCl ₃ as a white solid, 140.3 mg (82%) of (S)- 1- (6-Chloro-7-methoxyisoquinolin-3-yl) -3-((3-hydroxyquinuclidin-3-yl) methyl) thiourea was obtained.
LCMS RT 1.28 min, MH ⁺ = 407.1, 409.0, ¹ H NMR (500 MHz, chloroform-d) δ ppm 11.85 (1 H, t, J = 5.2 Hz), 9.15 (1 H, br.s.), 8.63 (1 H, s), 7.76 (1 H, s), 6.92 (1 H, s), 6.89 (1 H, s), 4.14 (1 H, dd, J = 13.9, 5.6 Hz), 3.99 (2 H , s), 3.90 (1 H, dd, J = 14.0, 5.2 Hz), 3.46 (3 H, s), 2.89-3.01 (2 H, m), 2.75-2.88 (3 H, m), 2.03-2.15 (1 H, m), 1.89-1.96 (1 H, m), 1.77-1.88 (1 H, m), 1.63 (1 H, ddd, J = 13.4, 6.1, 3.4 Hz), 1.32-1.45 (1 H , m).

ＤＭＦ(２.５ml)中の(Ｓ)−１−(６−クロロ−７−メトキシイソキノリン−３−イル)−３−((３−ヒドロキシキヌクリジン−３−イル)メチル)チオウレア(１４０mg, ０.３４４mmol)の溶液に、ＤＭＦ(０.５ml)中のＮ,Ｎ'−メタンジイリデンジプロパン−２−アミン(１０１mg, ０.８００mmol)の溶液を加え、得られた混合物を室温で１０日間静置した。溶媒を真空で蒸発させた。残渣を約５mlのＭｅＯＨに溶解し(幾らかの白色の固体を濾過して除いた, ０９)、分取ＨＰＬＣ精製を行い、次に１％ ＮＨ_４ＯＨ／９％ ＭｅＯＨ／９０％ ＣＨＣｌ_３でシリカゲルのクロマトグラフィーを行い、３１mg(２４％)の(Ｒ)−Ｎ−(６−クロロ−７−メトキシイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た。
LCMS RT 1.04分, MH⁺ = 373.2, 375.2, ¹H NMR (500 MHz, MeOD) δ ppm 8.78 - 8.89 (1 H, m), 7.90 - 8.01 (1 H, m), 7.14 - 7.23 (1 H, m), 4.01 (3 H, s), 3.95 - 3.99 (1 H, m), 3.62 - 3.71 (1 H, m), 3.19 - 3.25 (1 H, m), 3.06 - 3.13 (1 H, m), 2.90 - 3.00 (2 H, m), 2.76 - 2.89 (2 H, m), 2.10 - 2.21 (2 H, m), 1.69 - 1.83 (2 H, m), 1.56 - 1.68 (1 H, m)。 Step E: (R) -N- (6-Chloro-7-methoxyisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

(S) -1- (6-Chloro-7-methoxyisoquinolin-3-yl) -3-((3-hydroxyquinuclidin-3-yl) methyl) thiourea (140 mg, DMF (2.5 ml)) To a solution of 0.344 mmol) 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 stirred at room temperature for 10 minutes. Let stand for days. The solvent was evaporated in vacuo. The residue was dissolved in about 5 ml MeOH (some white solid was filtered off, 09) and preparative HPLC purification was performed followed by 1% NH ₄ OH / 9% MeOH / 90% CHCl ₃ . Chromatography on silica gel gave 31 mg (24%) of (R) -N- (6-chloro-7-methoxyisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine was obtained.
LCMS RT 1.04 min, MH ⁺ = 373.2, 375.2, ¹ H NMR (500 MHz, MeOD) δ ppm 8.78-8.89 (1 H, m), 7.90-8.01 (1 H, m), 7.14-7.23 (1 H, m), 4.01 (3 H, s), 3.95-3.99 (1 H, m), 3.62-3.71 (1 H, m), 3.19-3.25 (1 H, m), 3.06-3.13 (1 H, m) , 2.90-3.00 (2 H, m), 2.76-2.89 (2 H, m), 2.10-2.21 (2 H, m), 1.69-1.83 (2 H, m), 1.56-1.68 (1 H, m) .

Example 345
(R) -N- (5-Fluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

メタノール(１０ml)中の２,２−ジエトキシアセトイミド酸メチル(１.５ｇ, １０.２mmol)および(３−フルオロフェニル)メタンアミン(１.２７５ｇ, １０.２mmol)の混合物を、７０度で２時間撹拌した。溶媒を蒸発させて油状物である２,２−ジエトキシ−Ｎ−(３−フルオロベンジル)アセトイミドアミドを得た(２.５ｇ.１００％)。
¹H NMR (500 MHz, CDCl₃) δ 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, 酢酸アンモニウム系中) [M +H] = 254.7。 Step A: 7-Fluoroisoquinolin-3-amine and 5-Fluoroisoquinolin-3-amine

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) Stir for hours. The solvent was evaporated to give 2,2-diethoxy-N- (3-fluorobenzyl) acetimidamide as an oil (2.5 g. 100%).
¹ H NMR (500 MHz, CDCl ₃ ) δ 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.

To the oil prepared above (2.1 g) in CH ₂ Cl ₂ (3 ml) at room temperature was added 2 ml concentrated sulfuric acid very slowly and it was stirred at room temperature overnight. The mixture was poured into ice water and neutralized with NaOH (10N) to pH = 8. The mixture was extracted with EtOAc (100 ml x 3). The organic layers were combined, washed with water and brine, and dried over Na ₂ SO ₄ to give 1.05 g of a crude solid. This was used directly without further purification.

塩化メチレン(３０ml)中の上記の粗製の混合物(１０５０mg, ６.４７mmol)の溶液に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(１５００mg, ６.５mmol)を加えた。桃色の溶液を室温で２時間撹拌し、溶媒を蒸発させ、粗製の赤色の固体を得た。これをBiotageで精製し(０〜２５％ 酢酸エチル−ヘキサン)、７−フルオロ−３−イソチオシアナトイソキノリンと同定された最初のピークを単離した(６３５mg, ３.１１mmol, ４８％)。
¹H NMR (500 MHz, CDCl₃) δ ppm 9.08 (s, 1H), 7.9-7.8 (m, 1H), 7.7-7.6 (m, 1H), 7.6-7.5 (m, 2H)。 Step B: 5-Fluoro-3-isothiocyanatoisoquinoline

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 at room temperature for 2 hours and the solvent was evaporated to give a crude red solid. This was purified on Biotage (0-25% ethyl acetate-hexane) and the first peak identified as 7-fluoro-3-isothiocyanatoisoquinoline was isolated (635 mg, 3.11 mmol, 48%).
¹ H NMR (500 MHz, CDCl ₃ ) δ ppm 9.08 (s, 1H), 7.9-7.8 (m, 1H), 7.7-7.6 (m, 1H), 7.6-7.5 (m, 2H).

The second isomer, 5-fluoro-3-isothiocyanatoisoquinoline, was isolated (27.5 mg, 0.135 mmol, 2.1%).
¹ H NMR (500 MHz, CDCl ₃ ) δ 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).

ＤＭＦ(５ml)中の５−フルオロ−３−イソチオシアナトイソキノリン(２７.５mg, ０.１３５mmol)の溶液に、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(２５mg, ０.１６０mmol)および炭酸セシウム(１１０mg, ０.３３７mmol)を加えた。混合物を４０度で一夜撹拌した。反応物にＮ,Ｎ'−メタンジイリデンジプロパン−２−アミン(０.１２ｇ, ０.９５mmol)を加え、混合物を４０度で一夜撹拌した。溶媒を蒸発させ、粗製の固体をBiotageで精製し、最初に１００％ 酢酸エチルで、２番目のランで０〜２５％ ９：１ メタノール：水酸化アンモニウム−クロロホルムで、(Ｒ)−Ｎ−(５−フルオロイソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２９.４mg, ０.０８６mmol, ６４.１％)。
¹H NMR (500 MHz, MeOD) δ 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。 Step C: (R) -N- (5-Fluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine

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) were added. The mixture was stirred at 40 degrees overnight. To the reaction was added N, N′-methanediylidenedipropan-2-amine (0.12 g, 0.95 mmol) and the mixture was stirred at 40 ° C. overnight. The solvent was evaporated and the crude solid was purified by Biotage, first 100% ethyl acetate, 0-25% in the second run 9: 1 methanol: ammonium hydroxide-chloroform, (R) -N- ( 5-Fluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazol-5,3′-bicyclo [2.2.2] octane] -2-amine was obtained (29.4 mg, 0.086 mmol, 64.1%).
¹ H NMR (500 MHz, MeOD) δ 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'-bicyclo [2.2.2] octane] -2-amine

ＭｅＯＨ(３００ml)中のナトリウム メトキシド(１３.３８ｇ, ２４８mmol)の溶液に、２,２−ジエトキシアセトニトリル(１７.２２ml, １２４mmol)を滴下した。得られた混合物を室温で１６時間撹拌した。反応物を３００mlの水で希釈し、生成物を３×２５０mlのＥｔ_２Ｏで抽出した。合わせたＥｔ_２Ｏ層を１００mlの塩水で洗浄し、硫酸ナトリウムで乾燥させ、蒸発させ、望ましい生成物を無色の液体として得た(１５.３ｇ, ９５mmol, 収率７７％)。
¹H NMR (400 MHz, CDCl₃) δ ppm 7.86 (1 H, br. s.), 4.77 (1 H, s), 3.78 (3 H, s), 3.55 (4 H, qd, J=6.97, 4.28 Hz), 1.21 (7 H, t, J=7.05 Hz)。 Step A: Methyl 2,2-diethoxyacetimidate

To a solution of sodium methoxide (13.38 g, 248 mmol) in MeOH (300 ml) was added 2,2-diethoxyacetonitrile (17.22 ml, 124 mmol) dropwise. The resulting mixture was stirred at room temperature for 16 hours. The reaction was diluted with 300 ml water and the product was extracted with 3 × 250 ml Et ₂ O. The combined Et ₂ O layers were washed with 100 ml brine, dried over sodium sulfate and evaporated to give the desired product as a colorless liquid (15.3 g, 95 mmol, 77% yield).
¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.86 (1 H, br.s.), 4.77 (1 H, s), 3.78 (3 H, s), 3.55 (4 H, qd, J = 6.97, 4.28 Hz), 1.21 (7 H, t, J = 7.05 Hz).

ＴＨＦ(３０ml)中の２−ブロモ−５−フルオロベンゾニトリル(３.０ｇ, １５.００mmol)およびＮａＢＨ_４(１.４１９ｇ, ３７.５mmol)の溶液に、ＴＦＡ(３.４７ml, ４５.０mmol)を２０分かけてゆっくりと加えた。得られた混合物を室温で１６時間撹拌し、ＭｅＯＨ(１０ml)を加え、混合物をさらに３０分間撹拌した。それをＥｔＯＡｃ(２００ml)で希釈し、水で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、蒸発させた。残渣を８０ｇのThompsonシリカカートリッジで精製した(ヘキサン中３％〜１００％ Ｂ, １２００ml, Ｂ＝ＥｔＯＡｃ中１０％ ＭｅＯＨ)。望ましい生成物が、無色の油状物として得られた(１.７０ｇ, ８.３３mmol, 収率５５.５％)。
LC/MS (0.647分, MH⁺ = 205.92). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 7.59 (1 H, dd, J=8.85, 5.49 Hz), 7.43 (1 H, dd, J=10.07, 3.05 Hz), 7.05 (1 H, td, J=8.47, 3.20 Hz), 3.72 (3 H, s), 2.00 (2 H, br. s.)。 Step B: (2-Bromo-5-fluorophenyl) methanamine

To a solution of 2-bromo-5-fluorobenzonitrile (3.0 g, 15.00 mmol) and NaBH ₄ (1.419 g, 37.5 mmol) in THF (30 ml) was added TFA (3.47 ml, 45.0 mmol). Was added slowly over 20 minutes. The resulting mixture was stirred at room temperature for 16 hours, MeOH (10 ml) was added and the mixture was stirred for an additional 30 minutes. It was diluted with EtOAc (200 ml), washed with water, dried over Na ₂ SO ₄ and evaporated. The residue was purified on an 80 g Thompson silica cartridge (3% to 100% B in hexane, 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). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ ppm 7.59 (1 H, dd, J = 8.85, 5.49 Hz), 7.43 (1 H, dd, J = 10.07, 3.05 Hz), 7.05 (1 H, td, J = 8.47, 3.20 Hz), 3.72 (3 H, s), 2.00 (2 H, br. S.).

ＭｅＯＨ(２０ml)中の、２,２−ジエトキシアセトイミド酸メチル(１.２６４ｇ, ７.８４mmol)および(２−ブロモ−５−フルオロフェニル)メタンアミン(１.６ｇ, ７.８４mmol)の混合物を、７５℃で２時間撹拌した。溶媒を蒸発させ、薄黄色の固体として粗生成物を得た。このＮ−(２−ブロモ−５−フルオロベンジル)−２,２−ジエトキシアセトイミドアミド(２.６１ｇ, ７.８３mmol, 収率１００％)を、次の工程に直接用いた。
LC/MS (1.188分, MH⁺: 334.92). ¹H NMR (400 MHz, CDCl₃) δ ppm 7.44 (1 H, dd, J=8.69, 5.16 Hz), 7.15 (1 H, dd, J=9.44, 2.90 Hz), 6.81 (1 H, td, J=8.25, 3.15 Hz), 5.48 (1 H, br. s.), 4.90 (1 H, s), 4.42 (2 H, s), 3.49 - 3.67 (4 H, m), 1.17 - 1.25 (6 H, m)。 Step C: N- (2-bromo-5-fluorobenzyl) -2,2-diethoxyacetimidoamide

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). And stirred at 75 ° C. for 2 hours. The solvent was evaporated to give the crude product as a pale yellow solid. This N- (2-bromo-5-fluorobenzyl) -2,2-diethoxyacetimidamide (2.61 g, 7.83 mmol, 100% yield) was used directly in the next step.
LC / MS (1.188 min, MH ⁺ : 334.92). ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.44 (1 H, dd, J = 8.69, 5.16 Hz), 7.15 (1 H, dd, J = 9.44 , 2.90 Hz), 6.81 (1 H, td, J = 8.25, 3.15 Hz), 5.48 (1 H, br. S.), 4.90 (1 H, s), 4.42 (2 H, s), 3.49-3.67 (4 H, m), 1.17-1.25 (6 H, m).

Ｎ−(２−ブロモ−５−フルオロベンジル)−２,２−ジエトキシアセトイミドアミド(１.６ｇ, ４.８０mmol)およびジクロロメタン(２ml)の混合物に、Ｈ_２ＳＯ_４(１.２８０ml, ２４.０１mmol)を加えた。反応混合物を８０℃で１６時間撹拌し、室温まで冷却し、ＥｔＯＡｃ(１００ml)で希釈し、氷水で反応停止させ、ＮａＨＣＯ_３溶液で中和した。有機相を水で洗浄し、Ｎａ_２ＳＯ_４で乾燥させ、蒸発させた。残渣を直接８０ｇのThompsonシリカのカートリッジで精製した(ヘキサン中３％〜１００％ ＥｔＯＡｃ, １２００ml)。望ましい生成物を薄黄色の固体として得た(０.４７ｇ, １.９５０mmol, 収率４０.６％)。
LC/MS (1.052分, MH⁺: 242.99). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.95 (1 H, s), 7.36 (1 H, dd, J=8.06, 4.53 Hz), 7.23 (1 H, dd, J=10.58, 8.06 Hz), 6.68 (1 H, s), 6.52 (2 H, s), ¹⁹F NMR (376 MHz, DMSO-d₆) δ ppm -126.83 (1 F, s)。 Step D: 8-Bromo-5-fluoroisoquinolin-3-amine

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 ₂ SO ₄ (1.280 ml, 24 0.01 mmol) was added. The reaction mixture was stirred at 80 ° C. for 16 hours, cooled to room temperature, diluted with EtOAc (100 ml), quenched with ice water and neutralized with NaHCO ₃ solution. The organic phase was washed with water, dried over Na ₂ SO ₄ and evaporated. The residue was purified directly on an 80 g Thompson silica cartridge (3% to 100% EtOAc in hexane, 1200 ml). The desired product was obtained as a pale yellow solid (0.47 g, 1.950 mmol, 40.6% yield).
LC / MS (1.052 min, MH ⁺ : 242.99). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.95 (1 H, s), 7.36 (1 H, dd, J = 8.06, 4.53 Hz), 7.23 (1 H, dd, J = 10.58, 8.06 Hz), 6.68 (1 H, s), 6.52 (2 H, s), ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ ppm -126.83 (1 F , s).

８−ブロモ−５−フルオロイソキノリン−３−アミン(０.３０ｇ, １.２４５mmol)およびジクロロメタン(５ml)の混合物に、１,１'−チオカルボニルジピリジン−２(１Ｈ)−オン(０.３１８ｇ, １.３６９mmol)を加えた。得られた混合物を室温で３時間撹拌した。生成物を直接４０ｇのThompsonシリカのカートリッジで精製した(ヘキサン中３％〜１００％ ＥｔＯＡｃ, １２００ml)。望ましい生成物を灰白色の固体として得た(０.２３ｇ, ０.８１２mmol, 収率６５.３％)。
LC/MS (2.098分, MH⁺: 284.93). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.38 (1 H, s), 8.04 (1 H, dd, J=8.31, 4.78 Hz), 7.96 (1 H, s), 7.67 (1 H, dd, J=9.82, 8.31 Hz). ¹⁹F NMR (376 MHz, DMSO-d₆) δ ppm -122.13 (1 F, s)。 Step E: 8-Bromo-5-fluoro-3-isothiocyanatoisoquinoline

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 room temperature for 3 hours. The product was purified directly on a 40 g Thompson silica cartridge (3% to 100% EtOAc in hexane, 1200 ml). The desired product was obtained as an off-white solid (0.23 g, 0.812 mmol, 65.3% yield).
LC / MS (2.098 min, MH ⁺ : 284.93). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 9.38 (1 H, s), 8.04 (1 H, dd, J = 8.31, 4.78 Hz), 7.96 (1 H, s), 7.67 (1 H, dd, J = 9.82, 8.31 Hz). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ ppm -122.13 (1 F, s).

ＤＭＦ(５ml)中の、８−ブロモ−５−フルオロ−３−イソチオシアナトイソキノリン(０.２２ｇ, ０.７７７mmol)および(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(０.１９６ｇ, ０.８５５mmol)の混合物に、Ｃｓ_２ＣＯ_３(０.６３３ｇ, １.９４３mmol)を加えた。得られた混合物を４０℃で１６時間撹拌した。Ｎ,Ｎ'−メタンジイリデンジプロパン−２−アミン(０.２９４ｇ, ２.３２８mmol)を加え、４０℃でさらに１６時間撹拌を続けた。溶媒を蒸発させた。生成物を直接４０ｇのThompsonシリカのカートリッジで精製し(ヘキサン中３％〜１００％ Ｂ, １５００ml, Ｂ＝ＥｔＯＡｃ中２０％ ＭｅＯＨ)。生成物をさらにＥｔＯＡｃから再結晶した。望ましい生成物を灰白色の固体として得た(０.１１ｇ, ０.２６１mmol, 収率３３.６％)。
LC/MS (0.997分, MH⁺: 406.98). ¹H NMR (400 MHz, MeOD) δ ppm 9.30 (1 H, s), 7.60 (1 H, dd, J=8.18, 4.66 Hz), 7.22 (1 H, dd, J=10.07, 8.06 Hz), 4.00 (1 H, d, J=10.07 Hz), 3.69 (1 H, d, J=10.07 Hz), 3.23 (1 H, d, J=15.36 Hz), 3.07 - 3.13 (1 H, m), 2.90 - 2.96 (2 H, m), 2.76 - 2.85 (2 H, m), 2.15 (2 H, br. s.), 1.69 - 1.81 (2 H, m), 1.58 - 1.67 (1 H, m). ¹⁹F NMR (376 MHz, MeOD) δ ppm -126.72 (1 F, s)。 Step F: (R) -N- (8-Bromo-5-fluoroisoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2 -Amine

8-Bromo-5-fluoro-3-isothiocyanatoisoquinoline (0.22 g, 0.777 mmol) and (S) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride (5 ml) in DMF (5 ml). To a mixture of 0.196 g, 0.855 mmol) was added Cs ₂ CO ₃ (0.633 g, 1.943 mmol). The resulting mixture was stirred at 40 ° C. for 16 hours. N, N′-methanediylidenedipropan-2-amine (0.294 g, 2.328 mmol) was added and stirring was continued at 40 ° C. for a further 16 hours. The solvent was evaporated. The product was purified directly on a 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 an off-white solid (0.11 g, 0.261 mmol, 33.6% yield).
LC / MS (0.997 min, MH ⁺ : 406.98). ¹ H NMR (400 MHz, MeOD) δ ppm 9.30 (1 H, s), 7.60 (1 H, dd, J = 8.18, 4.66 Hz), 7.22 (1 H, dd, J = 10.07, 8.06 Hz), 4.00 (1 H, d, J = 10.07 Hz), 3.69 (1 H, d, J = 10.07 Hz), 3.23 (1 H, d, J = 15.36 Hz) , 3.07-3.13 (1 H, m), 2.90-2.96 (2 H, m), 2.76-2.85 (2 H, m), 2.15 (2 H, br. S.), 1.69-1.81 (2 H, m ), 1.58-1.67 (1 H, m). ¹⁹ F NMR (376 MHz, MeOD) δ ppm -126.72 (1 F, s).

Example 347
(R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- 2-Amine

４,６−ジクロロピリミジン(５.９６ｇ, ４０mmol)、イミダゾール(２.７２ｇ, ４０.０mmol)および炭酸カリウム(５.５３ｇ, ４０.０mmol)の混合物を、ＤＭＦ(５０ml)中で、室温で１８時間撹拌した。反応物を５００mlの水で希釈し、１５０mlのＥｔＯＡｃで５回抽出した。合わせた有機層を濃縮し、フラッシュクロマトグラフィーによって、２４０ｇのシリカゲルのカートリッジで、酢酸エチル中０〜１０％ メタノールで精製し、４−クロロ−６−(１Ｈ−イミダゾール−１−イル)ピリミジンを得た(４.５６ｇ, 収率６３％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.85 (1 H, d, J=0.76 Hz), 8.45 (1 H, s), 7.62 (1 H, t, J=1.51 Hz), 7.34 (1 H, d, J=0.76 Hz), 7.23 (1 H, dd, J=1.38, 0.88 Hz). LCMS: RT = 0.47分, MH⁺ = 181.1。 Step A: 4-Chloro-6- (1H-imidazol-1-yl) pyrimidine

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 added in DMF (50 ml) at room temperature for 18 hours. Stir for hours. The reaction was diluted with 500 ml water and extracted 5 times with 150 ml EtOAc. The combined organic layers were concentrated and purified by flash chromatography on a 240 g silica gel cartridge with 0-10% methanol in ethyl acetate to give 4-chloro-6- (1H-imidazol-1-yl) pyrimidine. (4.56 g, 63% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.85 (1 H, d, J = 0.76 Hz), 8.45 (1 H, s), 7.62 (1 H, t, J = 1.51 Hz), 7.34 (1 H, d, J = 0.76 Hz), 7.23 (1 H, dd, J = 1.38, 0.88 Hz). LCMS: RT = 0.47 min, MH ⁺ = 181.1.

４−クロロ−６−(１Ｈ−イミダゾール−１−イル)ピリミジン(４.０ｇ, ２２.１mmol)を、２つの等量の部分に分けて、メタノール中７Ｎ アンモニアの溶液(４０ml)と共に、２個の密封試験管に別々に入れた。試験管を密封し、６５℃で２０時間加熱し、室温で３日間静置した。反応物を合わせて、濃縮し、フラッシュクロマトグラフィーによって、２４０ｇのシリカゲルのカートリッジで、ヘキサン中４０〜１００％ ＥｔＯＡｃで、次にＥｔＯＡｃ中０〜５０％ ＭｅＯＨで精製し、６−(１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミンを得た(１.４３ｇ, ４０％)。
LCMS: RT = 0.39分, MH⁺ = 162.1。 Step B: 6- (1H-imidazol-1-yl) pyrimidin-4-amine

4-Chloro-6- (1H-imidazol-1-yl) pyrimidine (4.0 g, 22.1 mmol) was divided into two equal portions and 2 portions with a solution of 7N ammonia in methanol (40 ml). Were placed separately in sealed test tubes. The test tube was sealed, heated at 65 ° C. for 20 hours and 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 40-100% EtOAc in hexane, then 0-50% MeOH in EtOAc, and 6- (1H-imidazole- 1-yl) pyrimidin-4-amine was obtained (1.43 g, 40%).
LCMS: RT = 0.39 min, MH ⁺ = 162.1.

ＤＭＦ(１８ml)中の６−(１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミン(１.０ｇ, ６.２mmol)の溶液に、１０Ｍ ＮａＯＨ(１.２４ml, １２.４mmol)、二硫化炭素(０.９３ml, １６mmol)、ＮａＯＨ(１.２４ml, １２.４mmol)およびヨードメタン(０.９７ml, １６mmol)を１５分間で滴下した。２時間撹拌を続け、混合物を水に注いだ。濁った溶液をＥｔＯＡｃに分配し、水で洗浄し、濃縮し、フラッシュクロマトグラフィーによって、１１０ｇのシリカゲルのカートリッジで、ヘキサン中１０〜５０％ ＥｔＯＡｃで精製し、６−(１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチルを得た(４１１mg, 収率２５％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.79 (1 H, d, J=1.01 Hz), 8.65 (1 H, d, J=1.01 Hz), 7.96 (1 H, t, J=1.51 Hz), 7.33 (1 H, d, J=1.01 Hz), 7.12 - 7.19 (1 H, m), 2.58 (6 H, s). LCMS: RT = 0.63分, MH⁺ = 266.1。 Step C: Dimethyl 6- (1H-imidazol-1-yl) pyrimidin-4-ylcarbonimidodithioate

To a solution of 6- (1H-imidazol-1-yl) pyrimidin-4-amine (1.0 g, 6.2 mmol) in DMF (18 ml) was added 10M NaOH (1.24 ml, 12.4 mmol), carbon disulfide. (0.93 ml, 16 mmol), NaOH (1.24 ml, 12.4 mmol) and iodomethane (0.97 ml, 16 mmol) were added dropwise over 15 minutes. Stirring was continued for 2 hours and the mixture was poured into water. The cloudy solution was partitioned between EtOAc, washed with water, concentrated and purified by flash chromatography on a 110 g silica gel cartridge with 10-50% EtOAc in hexanes and 6- (1H-imidazol-1-yl. ) Dimethyl pyrimidin-4-ylcarbonimidodithioate was obtained (411 mg, 25% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.79 (1 H, d, J = 1.01 Hz), 8.65 (1 H, d, J = 1.01 Hz), 7.96 (1 H, t, J = 1.51 Hz), 7.33 (1 H, d, J = 1.01 Hz), 7.12-7.19 (1 H, m), 2.58 (6 H, s). LCMS: RT = 0.63 min, MH ⁺ = 266.1.

ＤＭＦ(３.８ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(４１９mg, １.８mmol)、６−(１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(４０４mg, １.５mmol)および炭酸セシウム(１.２４ｇ, ３.８mmol)の懸濁液を、７５℃で２時間撹拌した。反応物を濃縮し、フラッシュクロマトグラフィーによって、４０ｇのシリカゲルのカートリッジで、ＣＨＣｌ_３中０〜３％ [９：１ ＭｅＯＨ／ＮＨ_４ＯＨ]で精製し、(Ｒ)−Ｎ−(６−(１Ｈ−イミダゾール−１−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(４１７mg, 収率８３％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.40 (1 H, br. s.), 8.60 (1 H, d, J=1.01 Hz), 8.37 (1 H, t, J=1.01 Hz), 7.55 (1 H, t, J=1.39 Hz), 7.16 (1 H, dd, J=1.39, 0.88 Hz), 6.84 (1 H, br. s.), 3.98 (1 H, d, J=9.57 Hz), 3.64 (1 H, d, J=9.57 Hz), 3.34 (1 H, dd, J=14.98, 1.64 Hz), 2.69 - 3.04 (5 H, m), 2.08 - 2.21 (2 H, m), 1.67 - 1.78 (1 H, m), 1.43 - 1.62 (2 H, m). LCMS: RT = 0.26分, MH⁺ =326.2。 Step D: (R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (419 mg, 1.8 mmol), 6- (1H-imidazol-1-yl) pyrimidine-4-in DMF (3.8 ml) A suspension of dimethyl ylcarbonimidodithioate (404 mg, 1.5 mmol) and cesium carbonate (1.24 g, 3.8 mmol) was stirred at 75 ° C. for 2 hours. The reaction was concentrated and purified by flash chromatography on a 40 g silica gel cartridge with 0-3% [9: 1 MeOH / NH ₄ OH] in CHCl ₃ and (R) -N- (6- (1H -Imidazol-1-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazol-5,3'-bicyclo [2.2.2] octane] -2-amine was obtained (417 mg, yield). 83%).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.40 (1 H, br.s.), 8.60 (1 H, d, J = 1.01 Hz), 8.37 (1 H, t, J = 1.01 Hz), 7.55 (1 H, t, J = 1.39 Hz), 7.16 (1 H, dd, J = 1.39, 0.88 Hz), 6.84 (1 H, br. S.), 3.98 (1 H, d, J = 9.57 Hz ), 3.64 (1 H, d, J = 9.57 Hz), 3.34 (1 H, dd, J = 14.98, 1.64 Hz), 2.69-3.04 (5 H, m), 2.08-2.21 (2 H, m), 1.67-1.78 (1 H, m), 1.43-1.62 (2 H, 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 [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

４,６−ジクロロピリミジン(１１.９２ｇ, ８０mmol)、４−メチル−１Ｈ−イミダゾール(６.５７ｇ, ８０mmol)および炭酸セシウム(２６.１ｇ, ８０mmol)の混合物を、ＤＭＦ(５０ml)中で室温で１８時間撹拌した。反応物を２００mlの水に希釈し、２００mlのＥｔＯＡｃで３回抽出した。合わせた有機層を濃縮し、フラッシュクロマトグラフィーによって、３００ｇのシリカゲルのカートリッジで、ヘキサン中２５〜７５％ 酢酸エチルで精製し、２種の位置異性体生成物の混合物を得た。プールしたフラクションを、約２００mlの体積まで濃縮し、冷却し、４−クロロ−６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジンを得た(５.７０ｇ, 収率３７％)。そのＮＭＲ ＮＯＥの特性によって、４−メチルイミダゾール位置異性体の明確な同定を行った。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.80 (1 H, d, J=0.50 Hz), 8.36 (1 H, d, J=1.26 Hz), 7.29 - 7.31 (1 H, m), 7.25 (1 H, d, J=1.01 Hz), 2.27 (3 H, d, J=1.01 Hz). LCMS: RT = 0.50分, MH⁺ = 195.1。 Step A: 4-Chloro-6- (4-methyl-1H-imidazol-1-yl) pyrimidine

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 added in DMF (50 ml) at room temperature. Stir for 18 hours. The reaction was diluted in 200 ml water and extracted 3 times with 200 ml EtOAc. The combined organic layers were concentrated and purified by flash chromatography on a 300 g silica gel cartridge with 25-75% ethyl acetate in hexanes to give a mixture of two regioisomeric products. The pooled fractions were concentrated to a volume of about 200 ml and cooled to give 4-chloro-6- (4-methyl-1H-imidazol-1-yl) pyrimidine (5.70 g, 37% yield). The 4-methylimidazole positional isomer was clearly identified by the characteristics of its NMR NOE.
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.80 (1 H, d, J = 0.50 Hz), 8.36 (1 H, d, J = 1.26 Hz), 7.29-7.31 (1 H, m), 7.25 (1 H, d, J = 1.01 Hz), 2.27 (3 H, d, J = 1.01 Hz). LCMS: RT = 0.50 min, MH ⁺ = 195.1.

２Ｍ アンモニア(２０ml, ４０.０mmol)／イソプロパノール中の、４−クロロ−６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン(１.１８ｇ, ６.１mmol)の溶液を、密封バイアル中、８０℃で１８時間加熱した。冷却後、固体沈殿物を濾過し、乾燥させ、６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミンを得た(７３３mg, 収率６９.０％)。
¹H NMR (400 MHz, DMSO-d6) δ ppm 8.31 (1 H, d, J=1.26 Hz), 8.28 (1 H, d, J=0.76 Hz), 7.49 (1 H, t, J=1.13 Hz), 7.16 (2 H, s), 6.50 (1 H, d, J=0.76 Hz), 2.15 (3 H, d, J=0.76 Hz). LCMS: RT = 0.39分, MH⁺ = 176.1。 Step B: 6- (4-Methyl-1H-imidazol-1-yl) pyrimidin-4-amine

A solution of 4-chloro-6- (4-methyl-1H-imidazol-1-yl) pyrimidine (1.18 g, 6.1 mmol) in 2M ammonia (20 ml, 40.0 mmol) / isopropanol in a sealed vial. And heated at 80 ° C. for 18 hours. 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).
¹ H NMR (400 MHz, DMSO-d6) δ ppm 8.31 (1 H, d, J = 1.26 Hz), 8.28 (1 H, d, J = 0.76 Hz), 7.49 (1 H, t, J = 1.13 Hz) ), 7.16 (2 H, s), 6.50 (1 H, d, J = 0.76 Hz), 2.15 (3 H, d, J = 0.76 Hz). LCMS: RT = 0.39 min, MH ⁺ = 176.1.

ＤＭＦ(１２ml)中の６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミン(７００mg, ４.００mmol)の溶液に、１０Ｍ ＮａＯＨ(０.８ml, ８mmol)、二硫化炭素(０.６０ml, １０mmol)、ＮａＯＨ(０.８ml, ８mmol)およびヨードメタン(０.６２ml, １０mmol)を１５分間で滴下した。２時間撹拌を続け、混合物を水に注いだ。濁った溶液をＥｔＯＡｃで分配し、水で洗浄し、濃縮し、フラッシュクロマトグラフィーによって、１１０ｇのシリカゲルのカートリッジで、ヘキサン中５０〜１００％ ＥｔＯＡｃで精製し、６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチルを得た(４３８mg, 収率３９％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.76 (1 H, d, J=1.01 Hz), 8.55 (1 H, d, J=1.26 Hz), 7.62 - 7.66 (1 H, m), 7.24 (1 H, d, J=1.01 Hz), 2.57 (6 H, s), 2.24 (3 H, d, J=1.01 Hz). LCMS: RT = 0.66分, MH⁺ = 280.1。 Step C: 6- (4-Methyl-1H-imidazol-1-yl) pyrimidin-4-ylcarbonimidodithioate dimethyl

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 10M 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) were added dropwise over 15 minutes. Stirring was continued for 2 hours 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 50-100% EtOAc in hexanes and 6- (4-methyl-1H-imidazole. Dimethyl-1-yl) pyrimidin-4-ylcarbonimidodithioate was obtained (438 mg, 39% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.76 (1 H, d, J = 1.01 Hz), 8.55 (1 H, d, J = 1.26 Hz), 7.62-7.66 (1 H, m), 7.24 (1 H, d, J = 1.01 Hz), 2.57 (6 H, s), 2.24 (3 H, d, J = 1.01 Hz). LCMS: RT = 0.66 min, MH ⁺ = 280.1.

ＤＭＦ(３.８ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(４１２mg, １.８０mmol)、６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(４１９mg, １.５mmol)および炭酸セシウム(１.２２ｇ, ３.７５mmol)の懸濁液を、７５℃で２時間撹拌した。反応物を濃縮し、フラッシュクロマトグラフィーによって、４０ｇのシリカゲルのカートリッジで、ＥｔＯＡｃ中１％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]のプレラン、次にＣＨＣｌ_３中１〜２％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]で精製し、(Ｒ)−Ｎ−(６−(４−メチル−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(４７２mg, 収率９２％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.40 (1 H, br. s.), 8.56 (1 H, d, J=1.01 Hz), 8.28 (1 H, d, J=1.26 Hz), 7.21 (1 H, s), 6.76 (1 H, br. s.), 3.97 (1 H, d, J=9.57 Hz), 3.63 (1 H, d, J=9.57 Hz), 3.33 (1 H, dd, J=14.86, 1.76 Hz), 2.62 - 3.05 (5 H, m), 2.24 (3 H, d, J=1.01 Hz), 2.03 - 2.19 (2 H, m), 1.65 - 1.81 (1 H, m), 1.39 - 1.62 (2 H, m). LCMS: RT = 0.25, 0.46分, MH⁺ = 340.3。 Step D: (R) -N- (6- (4-Methyl-1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2. 2.2] Octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (412 mg, 1.80 mmol), 6- (4-methyl-1H-imidazol-1-yl) in DMF (3.8 ml) A suspension of dimethyl pyrimidin-4-ylcarbonimidodithioate (419 mg, 1.5 mmol) and cesium carbonate (1.22 g, 3.75 mmol) was stirred at 75 ° C. for 2 hours. The reaction was concentrated and flash chromatographed on a 40 g silica gel cartridge with a pre-run of 1% [95: 5 MeOH / NH ₄ OH] in EtOAc, then 1-2% in CHCl ₃ [95: 5 MeOH / NH ₄ OH] and purified by (R) -N- (6- (4-methyl-1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3 ′ -Bicyclo [2.2.2] octane] -2-amine was obtained (472 mg, 92% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.40 (1 H, br.s.), 8.56 (1 H, d, J = 1.01 Hz), 8.28 (1 H, d, J = 1.26 Hz), 7.21 (1 H, s), 6.76 (1 H, br.s.), 3.97 (1 H, d, J = 9.57 Hz), 3.63 (1 H, d, J = 9.57 Hz), 3.33 (1 H, dd, J = 14.86, 1.76 Hz), 2.62-3.05 (5 H, m), 2.24 (3 H, d, J = 1.01 Hz), 2.03-2.19 (2 H, m), 1.65-1.81 (1 H, m), 1.39-1.62 (2 H, 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 [oxazole-5,3′-bicyclo [2.2.2] ] Octane] -2-amine

４,６−ジクロロピリミジン(１.８ｇ, １２mmol)、４−クロロ−１Ｈ−イミダゾール(１.２３ｇ, １２.００mmol)および炭酸セシウム(３.９１ｇ, １２.００mmol)の混合物を、ＤＭＦ(８ml)中で、室温で１８時間撹拌した。反応物を水で希釈し、ＥｔＯＡｃで抽出した。合わせた有機層を濃縮し、フラッシュクロマトグラフィーによって、１２０ｇのシリカゲルのカートリッジで、ヘキサン中０〜４０％ 酢酸エチルで精製し、４−クロロ−６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジンを得た(１.４１ｇ, 収率５４.６％)。そのＮＭＲ ＮＯＥの特性によって、４−クロロイミダゾール位置異性体の明確な同定を行った。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.86 (1 H, d, J=0.76 Hz), 8.33 (1 H, d, J=1.51 Hz), 7.54 (1 H, d, J=1.76 Hz), 7.30 (1 H, d, J=0.76 Hz). LCMS: RT = 0.81分, MH⁺ = 215.1。 Step A: 4-Chloro-6- (4-chloro-1H-imidazol-1-yl) pyrimidine

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 added to DMF (8 ml). The mixture was stirred at room temperature for 18 hours. The reaction was diluted with water and extracted with EtOAc. The combined organic layers were concentrated and purified by flash chromatography on a 120 g silica gel cartridge with 0-40% ethyl acetate in hexane to give 4-chloro-6- (4-chloro-1H-imidazol-1-yl. ) Pyrimidine was obtained (1.41 g, yield 54.6%). The 4-chloroimidazole positional isomer was clearly identified by the characteristics of its NMR NOE.
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.86 (1 H, d, J = 0.76 Hz), 8.33 (1 H, d, J = 1.51 Hz), 7.54 (1 H, d, J = 1.76 Hz ), 7.30 (1 H, d, J = 0.76 Hz). LCMS: RT = 0.81 min, MH ⁺ = 215.1.

２Ｍ アンモニア(２０ml, ４０.０mmol)／イソプロパノール中の、４−クロロ−６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン(０.９９ｇ, ４.６０mmol)の溶液を、密封バイアル中、８０℃で１８時間加熱した。冷却後、固体沈殿物を濾過し、乾燥させ、６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミンを得た(８８４mg, 収率９８％)。
¹H NMR (400 MHz, DMSO-d6) δ ppm 8.44 (1 H, d, J=1.51 Hz), 8.32 (1 H, d, J=0.76 Hz), 7.96 (1 H, d, J=1.51 Hz), 7.30 (2 H, br. s.), 6.58 (1 H, d, J=1.01 Hz). LCMS: RT = 0.57分, MH⁺ = 196.1。 Step B: 6- (4-Chloro-1H-imidazol-1-yl) pyrimidin-4-amine

A solution of 4-chloro-6- (4-chloro-1H-imidazol-1-yl) pyrimidine (0.99 g, 4.60 mmol) in 2M ammonia (20 ml, 40.0 mmol) / isopropanol in a sealed vial. And heated at 80 ° C. for 18 hours. 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).
¹ H NMR (400 MHz, DMSO-d6) δ ppm 8.44 (1 H, d, J = 1.51 Hz), 8.32 (1 H, d, J = 0.76 Hz), 7.96 (1 H, d, J = 1.51 Hz) ), 7.30 (2 H, br. S.), 6.58 (1 H, d, J = 1.01 Hz). LCMS: RT = 0.57 min, MH ⁺ = 196.1.

ＤＭＦ(１２ml)中の６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン−４−アミン(８５０mg, ４.３５mmol)の溶液に、１０Ｍ ＮａＯＨ(０.８７ml, ８.７mmol)、二硫化炭素(０.６５ml, １０.９mmol)、ＮａＯＨ(０.８７ml, ８.７mmol)およびヨードメタン(０.６８ml, １０.９mmol)を１５分間で滴下した。２時間撹拌を続け、混合物を水に注いだ。濁った溶液をＥｔＯＡｃに分配し、水で洗浄し、濃縮し、フラッシュクロマトグラフィーによって、１１０ｇのシリカゲルのカートリッジで、ヘキサン中１０〜４０％ ＥｔＯＡｃで精製し、６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチルを得た(５５８mg, 収率４３％)。
回転異性体の約９：１混合物の¹H NMR (400 MHz, MeOD) δ ppm 8.80 (0.8 H, d, J=0.76 Hz), 8.79 (0.2 H, d, J=0.50 Hz), 8.59 (1 H, d, J=1.51 Hz), 8.50 (0.1 H, d, J=1.26 Hz), 7.97 (0.9 H, d, J=1.51 Hz), 7.84 (0.1 H, d, J=1.51 Hz), 7.32 (0.9 H, d, J=0.76 Hz), 2.63 (0.6 H, s), 2.58 (5.4 H, s). LCMS: RT = 0.94分, MH⁺ = 300.0。 Step C: Dimethyl 6- (4-chloro-1H-imidazol-1-yl) pyrimidin-4-ylcarbonimidodithioate

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 10 M NaOH (0.87 ml, 8.7 mmol), two Carbon sulfide (0.65 ml, 10.9 mmol), NaOH (0.87 ml, 8.7 mmol) and iodomethane (0.68 ml, 10.9 mmol) were added dropwise over 15 minutes. Stirring was continued for 2 hours and the mixture was poured into water. The cloudy solution was partitioned between EtOAc, washed with water, concentrated, and purified by flash chromatography on a 110 g silica gel cartridge with 10-40% EtOAc in hexanes to give 6- (4-chloro-1H-imidazole. Dimethyl -1-yl) pyrimidin-4-ylcarbonimidodithioate was obtained (558 mg, 43% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.80 (0.8 H, d, J = 0.76 Hz), 8.79 (0.2 H, d, J = 0.50 Hz), 8.59 (1 H, d, J = 1.51 Hz), 8.50 (0.1 H, d, J = 1.26 Hz), 7.97 (0.9 H, d, J = 1.51 Hz), 7.84 (0.1 H, d, J = 1.51 Hz), 7.32 (0.9 H, d, J = 0.76 Hz), 2.63 (0.6 H, s), 2.58 (5.4 H, s). LCMS: RT = 0.94 min, MH ⁺ = 300.0.

ＤＭＦ(３.８ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(４１２mg, １.８００mmol)、６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(４５０mg, １.５mmol)および炭酸セシウム(１.２２ｇ, ３.７５mmol)の懸濁液を、７５℃で２時間撹拌した。反応物を濃縮し、フラッシュクロマトグラフィーによって、４０ｇのシリカゲルのカートリッジで、ＥｔＯＡｃ中１％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]のプレラン、次にＣＨＣｌ_３中１〜２％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]で精製し、(Ｒ)−Ｎ−(６−(４−クロロ−１Ｈ−イミダゾール−１−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２６５mg, 収率４８％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.41 (1 H, br. s.), 8.60 (1 H, d, J=1.01 Hz), 8.25 (1 H, d, J=1.51 Hz), 7.44 (1 H, d, J=1.51 Hz), 6.78 (1 H, s), 4.00 (1 H, d, J=9.82 Hz), 3.67 (1 H, d, J=9.57 Hz), 3.36 (1 H, dd, J=14.86, 1.51 Hz), 2.73 - 3.06 (5 H, m), 2.09 - 2.22 (2 H, m), 1.69 - 1.79 (1 H, m), 1.46 - 1.64 (2 H, m). LCMS: RT = 0.54分, MH⁺ = 360.2。 Step D: (R) -N- (6- (4-Chloro-1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2. 2.2] Octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (412 mg, 1.800 mmol), 6- (4-chloro-1H-imidazol-1-yl) in DMF (3.8 ml) A suspension of dimethyl pyrimidin-4-ylcarbonimidodithioate (450 mg, 1.5 mmol) and cesium carbonate (1.22 g, 3.75 mmol) was stirred at 75 ° C. for 2 hours. The reaction was concentrated and flash chromatographed on a 40 g silica gel cartridge with a pre-run of 1% [95: 5 MeOH / NH ₄ OH] in EtOAc, then 1-2% in CHCl ₃ [95: 5 MeOH / NH ₄ OH] and purified by (R) -N- (6- (4-chloro-1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3 ′ -Bicyclo [2.2.2] octane] -2-amine was obtained (265 mg, 48% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.41 (1 H, br.s.), 8.60 (1 H, d, J = 1.01 Hz), 8.25 (1 H, d, J = 1.51 Hz), 7.44 (1 H, d, J = 1.51 Hz), 6.78 (1 H, s), 4.00 (1 H, d, J = 9.82 Hz), 3.67 (1 H, d, J = 9.57 Hz), 3.36 (1 H, dd, J = 14.86, 1.51 Hz), 2.73-3.06 (5 H, m), 2.09-2.22 (2 H, m), 1.69-1.79 (1 H, m), 1.46-1.64 (2 H, 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′-bicyclo [2.2.2] octane]- 2-Amine

４,６−ジクロロピリミジン(５.９６ｇ, ４０mmol)、１Ｈ−ピラゾール(２.７２ｇ, ４０.０mmol)および炭酸セシウム(１３.０３ｇ, ４０.０mmol)の混合物を、ＤＭＦ(２５ml)中で、室温で１８時間撹拌した。反応物を１００mlの水で希釈し、ＥｔＯＡｃで抽出した。合わせた有機層を濃縮し、フラッシュクロマトグラフィーによって、２４０ｇのシリカゲルのカートリッジで、ヘキサン中０〜２０％ 酢酸エチルで精製し、４−クロロ−６−(１Ｈ−ピラゾール−１−イル)ピリミジンを得た(４.８０ｇ, 収率６６％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.77 (1 H, d, J=0.76 Hz), 8.55 (1 H, d, J=2.77 Hz), 7.96 (1 H, d, J=1.01 Hz), 7.80 (1 H, d, J=1.01 Hz), 6.51 (1 H, dd, J=2.64, 1.64 Hz). LCMS: RT = 0.86分, MH⁺ = 181.1。 Step A: 4-Chloro-6- (1H-pyrazol-1-yl) pyrimidine

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) in DMF (25 ml) at room temperature. For 18 hours. The reaction was diluted with 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 0-20% ethyl acetate in hexanes to give 4-chloro-6- (1H-pyrazol-1-yl) pyrimidine. (4.80 g, 66% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 8.77 (1 H, d, J = 0.76 Hz), 8.55 (1 H, d, J = 2.77 Hz), 7.96 (1 H, d, J = 1.01 Hz ), 7.80 (1 H, d, J = 1.01 Hz), 6.51 (1 H, dd, J = 2.64, 1.64 Hz). LCMS: RT = 0.86 min, MH ⁺ = 181.1.

２Ｍ アンモニア(２０ml, ４０.０mmol)／イソプロパノール中の、４−クロロ−６−(１Ｈ−ピラゾール−１−イル)ピリミジン(１.３１ｇ, ７.２５mmol)の溶液を、密封バイアル中、８０℃で２４時間加熱した。反応物を室温で４日間保管した。固体沈殿物を濾過し、乾燥させ、６−(１Ｈ−ピラゾール−１−イル)ピリミジン−４−アミンを得た(１.１５ｇ, 収率９８％)。
¹H NMR (400 MHz, DMSO-d6) δ ppm 8.53 (1 H, dd, J=2.52, 0.50 Hz), 8.30 (1 H, d, J=0.76 Hz), 7.82 (1 H, d, J=1.01 Hz), 7.19 (2 H, br. s.), 6.88 (1 H, d, J=1.01 Hz), 6.55 (1 H, dd, J=2.64, 1.64 Hz). LCMS: RT = 0.52分, MH⁺ = 162.1。 Step B: 6- (1H-pyrazol-1-yl) pyrimidin-4-amine

A solution of 4-chloro-6- (1H-pyrazol-1-yl) pyrimidine (1.31 g, 7.25 mmol) in 2M ammonia (20 ml, 40.0 mmol) / isopropanol at 80 ° C. in a sealed vial. Heated for 24 hours. 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).
¹ H NMR (400 MHz, DMSO-d6) δ ppm 8.53 (1 H, dd, J = 2.52, 0.50 Hz), 8.30 (1 H, d, J = 0.76 Hz), 7.82 (1 H, d, J = 1.01 Hz), 7.19 (2 H, br. S.), 6.88 (1 H, d, J = 1.01 Hz), 6.55 (1 H, dd, J = 2.64, 1.64 Hz). LCMS: RT = 0.52 min, MH ⁺ = 162.1.

ＤＭＦ(１８ml)中の６−(１Ｈ−ピラゾール−１−イル)ピリミジン−４−アミン(１.０ｇ, ６.２０mmol)の溶液に、１０Ｍ ＮａＯＨ(１.２４ml, １２.４mmol)、二硫化炭素(０.９３３ml, １５.５mmol)、ＮａＯＨ(１.２４ml, １２.４mmol)およびヨードメタン(０.９６６ml, １５.５mmol)を１５分間で滴下した。一夜撹拌を続け、混合物を水に注いだ。黄褐色の沈殿物を濾過し、水で洗浄し、乾燥させ、６−(１Ｈ−ピラゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチルを得た(１３６mg, 収率８％)。
¹H NMR (400 MHz, MeOD) δ ppm 8.74 (1 H, d, J=1.01 Hz), 8.64 (1 H, d, J=2.52 Hz), 7.82 (1 H, d, J=1.26 Hz), 7.40 (1 H, d, J=1.01 Hz), 6.57 (1 H, dd, J=2.64, 1.64 Hz), 2.58 (6 H, s). LCMS: RT = 0.96分, MH⁺ = 266.1。 Step C: Dimethyl 6- (1H-pyrazol-1-yl) pyrimidin-4-ylcarbonimidodithioate

To a solution of 6- (1H-pyrazol-1-yl) pyrimidin-4-amine (1.0 g, 6.20 mmol) in DMF (18 ml) was added 10M 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) were added dropwise over 15 minutes. Stirring was continued overnight and the mixture was poured into water. The tan precipitate was filtered, washed with water and dried to give dimethyl 6- (1H-pyrazol-1-yl) pyrimidin-4-ylcarbonimidodithioate (136 mg, 8% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 8.74 (1 H, d, J = 1.01 Hz), 8.64 (1 H, d, J = 2.52 Hz), 7.82 (1 H, d, J = 1.26 Hz), 7.40 (1 H, d, J = 1.01 Hz), 6.57 (1 H, dd, J = 2.64, 1.64 Hz), 2.58 (6 H, s). LCMS: RT = 0.96 min, MH ⁺ = 266.1.

ＤＭＦ(１.２ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(１３５mg, ０.５９mmol)、６−(１Ｈ−ピラゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(１３０mg, ０.４９mmol)および炭酸セシウム(３９９mg, １.２３mmol)の懸濁液を、７５℃で２時間撹拌した。反応物を濃縮し、フラッシュクロマトグラフィーによって、４０ｇのシリカゲルのカートリッジで、ＥｔＯＡｃ中１％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]のプレランで、次に定組成のＣＨＣｌ_３中１％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]で精製し、(Ｒ)−Ｎ−(６−(１Ｈ−ピラゾール−１−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(１０５mg, 収率６６％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.46 (1 H, br. s.), 8.58 (1 H, d, J=1.01 Hz), 8.50 (1 H, dd, J=2.64, 0.63 Hz), 7.74 (1 H, d, J=1.01 Hz), 7.47 (1 H, br. s.), 6.44 (1 H, dd, J=2.64, 1.64 Hz), 3.98 (1 H, d, J=9.57 Hz), 3.65 (1 H, d, J=9.57 Hz), 3.39 (1 H, dd, J=14.86, 1.51 Hz), 2.71 - 3.09 (5 H, m), 2.10 - 2.26 (2 H, m), 1.69 - 1.80 (1 H, m), 1.48 - 1.64 (2 H, m). LCMS: RT = 0.51分, MH⁺ = 326.2。 Step D: (R) -N- (6- (1H-pyrazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (135 mg, 0.59 mmol), 6- (1H-pyrazol-1-yl) pyrimidine-4-in DMF (1.2 ml) A suspension of dimethyl ylcarbonimidodithioate (130 mg, 0.49 mmol) and cesium carbonate (399 mg, 1.23 mmol) was stirred at 75 ° C. for 2 hours. The reaction was concentrated and flash chromatographed on a 40 g silica gel cartridge with a pre-run of 1% [95: 5 MeOH / NH ₄ OH] in EtOAc, then 1% in isocratic CHCl ₃ [95: 5 Purified with MeOH / NH ₄ OH] and (R) —N- (6- (1H-pyrazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] Octane] -2-amine was obtained (105 mg, 66% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.46 (1 H, br.s.), 8.58 (1 H, d, J = 1.01 Hz), 8.50 (1 H, dd, J = 2.64, 0.63 Hz ), 7.74 (1 H, d, J = 1.01 Hz), 7.47 (1 H, br. S.), 6.44 (1 H, dd, J = 2.64, 1.64 Hz), 3.98 (1 H, d, J = 9.57 Hz), 3.65 (1 H, d, J = 9.57 Hz), 3.39 (1 H, dd, J = 14.86, 1.51 Hz), 2.71-3.09 (5 H, m), 2.10-2.26 (2 H, m ), 1.69-1.80 (1 H, m), 1.48-1.64 (2 H, 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] Octane] -2-amine

４,６−ジクロロピリミジン(５.９６ｇ, ４０mmol)、１Ｈ−１,２,４−トリアゾール(１.３８１ｇ, ２０.００mmol)および炭酸セシウム(１３.０３ｇ, ４０.０mmol)の混合物を、ＤＭＦ(２５ml)中で、室温で１６時間撹拌した。反応物を１００mlの水で希釈し、ＥｔＯＡｃで抽出した。合わせた有機抽出物を濃縮し、フラッシュクロマトグラフィーによって、１６０ｇのシリカゲルカートリッジで、ヘキサン中２５〜５０％ ＥｔＯＡｃで精製し、４−クロロ−６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジンを得た(２.７８ｇ, 収率３８％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.21 (1 H, s), 8.87 (1 H, s), 8.15 (1 H, s), 7.91 (1 H, s). LCMS: RT = 0.68分, MH⁺ = 182.1。 Step A: 4-Chloro-6- (1H-1,2,4-triazol-1-yl) pyrimidine

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 added to DMF ( 25 ml) at room temperature for 16 hours. 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 25-50% EtOAc in hexanes to give 4-chloro-6- (1H-1,2,4-triazole-1- Yl) pyrimidine was obtained (2.78 g, 38% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.21 (1 H, s), 8.87 (1 H, s), 8.15 (1 H, s), 7.91 (1 H, s). LCMS: RT = 0.68 Minutes, MH ⁺ = 182.1.

２Ｍ アンモニア(２０ml, ４０.０mmol)／イソプロパノール中の４−クロロ−６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン(１.２４ｇ, ６.８３mmol)の溶液を、８０℃で、密封バイアル中で、１８時間加熱した。冷却後、固体沈殿物を濾過し、乾燥させ、６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン−４−アミンを得た(１.０６ｇ, 収率９６％)。
¹H NMR (400 MHz, DMSO-d6) δ ppm 9.30 (1 H, s), 8.35 (1 H, s), 8.30 (1 H, s), 7.39 (2 H, br. s.), 6.83 (1 H, d, J=0.76 Hz). LCMS: RT = 0.41分, MH⁺ = 163.1。 Step B: 6- (1H-1,2,4-triazol-1-yl) pyrimidin-4-amine

A solution of 4-chloro-6- (1H-1,2,4-triazol-1-yl) pyrimidine (1.24 g, 6.83 mmol) in 2M ammonia (20 ml, 40.0 mmol) / isopropanol was added at 80 ° C. And heated in a sealed vial for 18 hours. 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).
¹ H NMR (400 MHz, DMSO-d6) δ ppm 9.30 (1 H, s), 8.35 (1 H, s), 8.30 (1 H, s), 7.39 (2 H, br.s.), 6.83 ( 1 H, d, J = 0.76 Hz). LCMS: RT = 0.41 min, MH ⁺ = 163.1.

ＤＭＦ(１８ml)中の、６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン−４−アミン(１.００ｇ, ６.１７mmol)の溶液に、１０Ｍ ＮａＯＨ(１.２３ml, １２.３mmol)、二硫化炭素(０.９２７ml, １５.４mmol)、ＮａＯＨ(１.２３ml, １２.３mmol)およびヨードメタン(０.９６ml, １５mmol)を１５分間で滴下した。２時間撹拌を続け、混合物を水に注いだ。濁った溶液をＥｔＯＡｃで分配し、水で洗浄し、濃縮し、１１０ｇのシリカゲルのカートリッジで、ヘキサン中１０〜５０％ ＥｔＯＡｃで、フラッシュクロマトグラフィーによって精製し、６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチルを得た(５４１mg, 収率３３％)。
回転異性体の４：１混合物の¹H NMR (400 MHz, MeOD) δ ppm 9.42 (0.74 H, s), 9.40 (0.35 H, s), 9.04 (0.32 H, s), 8.84 (0.76 H, d, J=1.01 Hz), 8.81 (0.33 H, d, J=1.01 Hz), 8.23 (1 H, s), 7.42 (0.75 H, d, J=1.01 Hz), 2.64 (1.2 H, s), 2.59 (4.8 H, s). LCMS: RT = 0.86分, MH⁺ = 267.1。 Step C: 6- (1H-1,2,4-triazol-1-yl) pyrimidin-4-ylcarbonimidodithioic acid dimethyl ester

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 10M 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) were added dropwise over 15 minutes. Stirring was continued for 2 hours and the mixture was poured into water. The cloudy solution was partitioned with EtOAc, washed with water, concentrated, purified by flash chromatography on a 110 g silica gel cartridge with 10-50% EtOAc in hexanes, and 6- (1H-1,2,4 -Triazol-1-yl) pyrimidin-4-ylcarbonimidodithioate dimethyl was obtained (541 mg, 33% yield).
¹ H NMR (400 MHz, MeOD) δ ppm 9.42 (0.74 H, s), 9.40 (0.35 H, s), 9.04 (0.32 H, s), 8.84 (0.76 H, d) of a 4: 1 mixture of rotamers , J = 1.01 Hz), 8.81 (0.33 H, d, J = 1.01 Hz), 8.23 (1 H, s), 7.42 (0.75 H, d, J = 1.01 Hz), 2.64 (1.2 H, s), 2.59 (4.8 H, s). LCMS: RT = 0.86 min, MH ⁺ = 267.1.

ＤＭＦ(３.８ml)中の、(Ｓ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩(４１２mg, １.８００mmol)、６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン−４−イルカルボンイミドジチオ酸ジメチル(４００mg, １.５mmol)および炭酸セシウム(１２２２mg, ３.７５mmol)の懸濁液を、７５℃で２時間撹拌した。反応物を濃縮し、４０ｇのシリカゲルのカートリッジで、ＥｔＯＡｃ中１％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]のプレランで、次にＣＨＣｌ_３中１〜３％ [９５：５ ＭｅＯＨ／ＮＨ_４ＯＨ]で、フラッシュクロマトグラフィーによって精製し、(Ｒ)−Ｎ−(６−(１Ｈ−１,２,４−トリアゾール−１−イル)ピリミジン−４−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを得た(２１０mg, 収率４３％)。
¹H NMR (400 MHz, クロロホルム-d) δ ppm 9.45 (1 H, br. s.), 9.14 (1 H, s), 8.61 (1 H, d, J=1.01 Hz), 8.08 (1 H, s), 7.39 (1 H, s), 4.00 (1 H, d, J=9.57 Hz), 3.67 (1 H, d, J=9.57 Hz), 3.38 (1 H, dd, J=14.86, 1.76 Hz), 2.71 - 3.08 (5 H, m), 2.08 - 2.23 (2 H, m), 1.68 - 1.80 (1 H, m), 1.45 - 1.64 (2 H, m). LCMS: RT = 0.46分, MH⁺ = 327.2。 Step D: (R) -N- (6- (1H-1,2,4-triazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [ 2.2.2] Octane] -2-amine

(S) -3- (Aminomethyl) quinuclidin-3-ol dihydrochloride (412 mg, 1.800 mmol), 6- (1H-1,2,4-triazole-1-) in DMF (3.8 ml) A suspension of dimethyl) pyrimidin-4-ylcarbonimidodithioate (400 mg, 1.5 mmol) and cesium carbonate (1222 mg, 3.75 mmol) was stirred at 75 ° C. for 2 hours. The reaction was concentrated and a 40 g silica gel cartridge with a pre-run of 1% [95: 5 MeOH / NH ₄ OH] in EtOAc, then 1-3% in CHCl ₃ [95: 5 MeOH / NH ₄ OH]. And purified by flash chromatography and (R) -N- (6- (1H-1,2,4-triazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5 , 3′-bicyclo [2.2.2] octane] -2-amine was obtained (210 mg, 43% yield).
¹ H NMR (400 MHz, chloroform-d) δ ppm 9.45 (1 H, br.s.), 9.14 (1 H, s), 8.61 (1 H, d, J = 1.01 Hz), 8.08 (1 H, s), 7.39 (1 H, s), 4.00 (1 H, d, J = 9.57 Hz), 3.67 (1 H, d, J = 9.57 Hz), 3.38 (1 H, dd, J = 14.86, 1.76 Hz ), 2.71-3.08 (5 H, m), 2.08-2.23 (2 H, m), 1.68-1.80 (1 H, m), 1.45-1.64 (2 H, m). LCMS: RT = 0.46 min, MH ⁺ = 327.2.

(Ｓ)−Ｎ−(イソキノリン−３−イル)−４Ｈ−１'−アザスピロ[オキサゾール−５,３'−ビシクロ[２.２.２]オクタン]−２−アミンを、実施例２３９(工程Ａ〜Ｂ)の方法によって、(Ｒ)−３−(アミノメチル)キヌクリジン−３−オール 二塩酸塩から出発して製造した。
¹H NMR (400 MHz, MeOD) δ ppm 9.00 (1 H, s), 7.91 (1 H, d, J=8.06 Hz), 7.70 (1 H, d, J=8.31 Hz), 7.58 (1 H, t, J=7.18 Hz), 7.40 (1 H, t, J=7.05 Hz), 7.30 (1 H, br. s.), 3.95 (1 H, d, J=9.82 Hz), 3.64 (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=7.30 Hz), 2.80 (2 H, t, J=7.05 Hz), 2.05 - 2.23 (2 H, m), 1.52 - 1.84 (3 H, m). MS (LC/MS) R.T. =1.39; [M+H]⁺= 309.21。 Example 352
(S) -N- (isoquinolin-3-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2.2.2] octane] -2-amine

(S) -N- (isoquinolin-3-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane] -2-amine was prepared according to Example 239 (Step A Prepared starting from (R) -3- (aminomethyl) quinuclidin-3-ol dihydrochloride by the method of ~ B).
¹ H NMR (400 MHz, MeOD) δ ppm 9.00 (1 H, s), 7.91 (1 H, d, J = 8.06 Hz), 7.70 (1 H, d, J = 8.31 Hz), 7.58 (1 H, t, J = 7.18 Hz), 7.40 (1 H, t, J = 7.05 Hz), 7.30 (1 H, br.s.), 3.95 (1 H, d, J = 9.82 Hz), 3.64 (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 = 7.30 Hz), 2.80 (2 H, t, J = 7.05 Hz), 2.05-2.23 (2 H, m), 1.52-1.84 (3 H, m). MS (LC / MS) RT = 1.39; [M + H] ⁺ = 309.21.

  It will be apparent to those skilled in the art that the present disclosure is not limited to the embodiments described above and can be embodied in other specific forms without departing from the essential characteristics thereof. Accordingly, the embodiments are illustrative in all respects, and should be construed as being non-limiting, referring to the claims rather than the foregoing embodiments, and fall within the meaning and scope equivalent to the claims. All changes are intended to be included in it.

Claims (9)

Formula Ia:

[Wherein R ¹ is pyrimidinyl substituted with one imidazolyl]
Or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. (R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1′-azaspiro [oxazole-5,3′-bicyclo [2.2.2] octane]- 2-Amine:

Or a pharmaceutically acceptable salt thereof.   A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. The compound of claim 1 is (R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2. 2.2] Octane] -2-amine:

Or the pharmaceutical composition of Claim 3 which is a pharmaceutically acceptable salt thereof.   An agent for treating schizophrenia, Alzheimer's disease, cognitive impairment, rheumatoid arthritis, osteoarthritis, ulcerative colitis, Crohn's disease or diabetes comprising the compound of claim 1.   6. The agent of claim 5 for treating schizophrenia. The compound of claim 1 is (R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2. 2.2] Octane] -2-amine:

Or the agent of Claim 6 which is its pharmaceutically acceptable salt.   The agent of claim 5 for treating Alzheimer's disease. The compound of claim 1 is (R) -N- (6- (1H-imidazol-1-yl) pyrimidin-4-yl) -4H-1'-azaspiro [oxazole-5,3'-bicyclo [2. 2.2] Octane] -2-amine:

Or the agent of Claim 8 which is the pharmaceutically acceptable salt thereof.