KR100437973B1 - Isoxazole and Isoxazoline Substituted 1,2,5,6-Tetrahydropyridine Derivatives Acting on Acetylcholinic Receptor - Google Patents

Abstract

The present invention is a muscarinic acetylcholine receptor agonist, isooxazole or isooxazolin substituted 1,2,5,6-tetrahydropyridine compound of Formula 1 useful as a recognition enhancer and a therapeutic agent for cerebral neuropathy, and a pharmaceutical agent thereof Scientifically acceptable salts, methods for their preparation, and pharmaceutical compositions containing the same.

In the above formula,

R ² represents C _1-4 -alkyl,

One of R ³ and R ⁴ , The other is H,

R ⁵ is hydrogen, hydroxy, hydroxy-C _1-6 -alkyl, C _{1-6 -alkoxymethyl} , C _1-6 -alkoxy, acetoxy, C _1-4 -alkyl ester, nitrile, aryl, phenylthio , Methylthiomethyl, phenylsulfoxide, dimethylthiomethyl, C _1-4 -alkylketo, C _1-4 -alkylketooxime, C _1-4 -alkylketo-C _1-5 -alkyloxime, N-pyrroli Din-2-one,

R ⁶ is hydrogen,

R ⁵ and R ⁶ together form —CH ₂ OCH ₂ —.

Description

Isoxazole and Isoxazoline Substituted 1,2,5,6-Tetrahydropyridine Derivatives Acting on Acetylcholinic Receptor that Acts on Acetylcholine Receptor Substituted 1,2,5,6-tetrahydropyridine Derivatives

The present invention relates to isoxoxazole or isoxoxazol substituted 1,2,5,6-tetrahydropyridine compounds and pharmaceutically acceptable compounds which are useful as muscarinic acetylcholine receptor agonists, and are useful as recognition enhancers and therapeutic agents for brain neurological diseases. Salts, methods for their preparation, and pharmaceutical compositions containing the same.

As the elderly population grows, senile diseases including dementia are increasing. Geriatric dementia disease, represented by Alzheimer's disease, is a neurodegenerative disorder that presents symptoms of intellectual disability such as memory, judgment, and cognitive function. In Alzheimer's disease, muscarinic acetylcholine neurons in the presynaptic cerebral basal ganglia projected to the frontal lobe and hippocampus involved in cognitive functions such as learning, association, hardening, and cognition degenerate up to 90%, but postsynaptic muscarinogenicity Neurons are relatively unchanged. This suggests a strategy for drug development based on the cholinergic deficiency hypothesis focused on stimulation of postsynaptic receptors [R.T. Bartus, et al. Science, 217, 408-417 (1982).

As an acetylcholine esterase inhibitor to increase useful acetylcholine, tacrine has been developed as a drug related to cognitive ability, but has side effects, and recently, aricept (aricept, donepezil, Eisai America, Inc., 1996), Exelon (Ribastigmine, Novartis Pharmaceuticals Corporation, 2000), Reminyl (galantamine hydrobromide, Janssen Research, 2001) have been developed [W. Greenlee, et al. Il Farmaco, 2001, 56, 247-250]. However, oxotremorine, RS-86 (RS-86), which are nonselective cholinergic agonists for direct stimulation of cholinergic receptors, had side effects [R. Plate et al., Bioorg. Med. Chem., 2000, 8, 449-454.

Muscarinic choline receptors have five subtypes in the central and peripheral nervous systems and play an important role in cognitive action. It is known that post-synaptic muscarinic neurons in the brain and hippocampus remain relatively unchanged in patients with Alzheimer's disease. The research focuses on the development of therapeutics and cognitive enhancers. In addition, muscarinic receptors, which play an important role in the learning and memory functions that exist before and after synapses in the cholinergic neurotransmitter system, regulate the progression of amyloid precursor proteins involved in beta amyloid precipitation in Alzheimer's patients, and muscarinic receptor efficacy Since it is known to promote the secretion of soluble amyloid precursor protein and to reduce phosphorylation of tau-protein, high drug and low cholinergic properties for the development of therapeutic agent and cognitive enhancer for Alzheimer's disease where beta amyloid plaque and nerve fiber entanglement accumulate It is important to develop muscarinic receptor agonists with side effects and selective new muscarinic acetylcholine receptor activity to other receptors [C. C. Felder et al., J. Med. Chem. 2000, 43, 23, 4334-4353.

Muscarinic agonists that are active in the central nervous system have been published: Talsaclidin (1997), YM-796 (YM-796, 1990), CI-1017 (CI-1017, 2000), Xanomelin , 1997), milameline (Milameline, 1997), sabcomelin (Sabcomeline, SB-202026, 1997), albameline (Alvameline, LU 25-109, 1997), AF-102 (AF-102, 1997), etc. [A. Fisher, Drug Dev. Res. 2000, 50, 291-297. In addition, oxadiazole compounds with high affinity and good efficacy have been reported but side effects have been announced. Recently, muscarinic agonists, pilocarpine (Pilocarpine, Salagen Tablets, MGI Pharma, Inc., 1998) and Sevimeline (Cevimeline, AF102B, EVOXAC ^™ , SnowBrand Pharmaceuticals, Inc., 2000) have been shown to affect autoimmune glands It is approved by the FDA for the treatment of dry mouth in patients with Scheren's disease (Drugs of the future 2000, 25 (6), 558-562).

However, compounds active on muscarinic acetylcholine receptors are useful for pain, glaucoma, schizophrenia, anxiety, manic depression (circulatory psychosis), bipolar psychosis, depression, sleep disorders, epilepsy, cerebral ischemia, bowel incontinence, and gastrointestinal disorders. [L. M. Merritt et al., US Pat. No. 5,998,404. Some compounds that are active on the muscarinic acetylcholine receptor have side effects such as excessive saliva, tears, and gastrointestinal disorders. Therefore, there is still a need for the development of compounds with new muscarinic acetylcholine receptor activity that exhibit high efficacy and low cholinergic side effects.

Accordingly, it is an object of the present invention to have 1,2,5,6-tetrahydropyridine and isooxazole and isoxazoline derivatives substituted thereof having a high affinity for muscarinic acetylcholine receptors and thus useful as recognition enhancers and therapeutic agents for neurological diseases. To provide a pharmaceutically acceptable salt, a method for preparing the same, and a pharmaceutical composition containing the same.

The present invention relates to substituted 1,2,5,6-tetrahydropyridine compounds of the formula (1) and their pharmaceutically acceptable salts, and methods for their preparation.

<Formula 1>

In the above formula,

R ² represents C _1-4 -alkyl,

One of R ³ and R ⁴ , The other is H,

R ⁵ is hydrogen, hydroxy, hydroxy-C _1-6 -alkyl, C _{1-6 -alkoxymethyl} , C _1-6 -alkoxy, acetoxy, C _1-4 -alkyl ester, nitrile, aryl, phenylthio , Methylthiomethyl, phenylsulfoxide, dimethylthiomethyl, C _1-4 -alkylketo, C _1-4 -alkylketooxime, C _1-4 -alkylketo-C _1-5 -alkyloxime, N-pyrroli Din-2-one,

R ⁶ is hydrogen,

R ⁵ and R ⁶ together form —CH ₂ OCH ₂ —.

As used herein, the terms "C _1-4 -alkyl", "C _1-5 -alkyl" and "C _1-6 -alkyl" each represent 1 to 4, 1 to 5, and 1 to 6 carbon atoms. It means a dog-containing straight or branched chain alkyl. Examples of the alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl and the like, with methyl or ethyl being preferred.

As used herein, the term “C _1-6 -alkoxy” means straight or branched chain alkoxy containing 1 to 6 carbon atoms. For example, there are methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy and the like, with methoxy or ethoxy being preferred.

The term "aryl" as used herein denotes phenyl, vinyl, naphthyl, with phenyl being preferred.

Acids which form pharmaceutically acceptable acid addition salts include hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, acetic acid, benzoic acid, sulfonic acid, fumaric acid, maleic acid, citric acid, lactic acid, tartaric acid, oxalic acid or similar pharmaceutically acceptable organic and inorganic acids. And the like.

Preferred examples of the compound of Formula 1 according to the present invention are

[3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -methanol,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanol,

1-methyl-3- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) isoxazole-5-carboxylic acid methyl ester,

1-methyl-3- (5-phenyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone oxime,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone O-methyl oxime,

[3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -methanol,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanol,

1-methyl-4- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

3- (1-methyl-1,2,5,6-tetrahydro-pyridin-4-yl) isoxazole-5-carboxylic acid methyl ester,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone oxime,

1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone O-methyl oxime,

1-methyl-3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

1-methyl-3- (5-butoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-ol,

1-methyl-3- (isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

1-methyl-3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

[3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methanol,

1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -ethanol,

3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazole-5-carbonitrile,

1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -pyrrolidin-2-one ,

1-methyl-3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine,

3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -3a, 4,6,6a-tetrahydro-puro [3,4-d] isoxazole,

1-methyl-3- (5-methylsulfanylmethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, or

Dimethyl- [3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methyl-sulfonium iodide Droid,

And pharmaceutically acceptable salts thereof.

In addition, the present invention is to obtain a nitrile oxide of the formula (4) by using the 3- or 4-pyridine aldehyde oxime compound of the formula (2) as a starting material, and by ring addition reaction with an alkene or alkyne compound of the formula (8) or (9) Preparation of the compound of Formula 1, comprising preparing a compound of Formula 5 or 6 and reacting the compound of Formula 5 or 6 with alkyl iodide to prepare an alkyl pyridine salt of Formula 7, and then reducing the compound Provide a method.

<Formula 1>

In the formula, R ¹ represents 3- or 4-pyridine, and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above.

The method for preparing the compound of Formula 1 of the present invention will be described in more detail with reference to Schemes 1 and 2 below.

First, as described in Scheme 1, a 3- or 4-pyridine aldehyde oxime compound of formula (2) and N-chloro succinimide are solvents such as tetrahydrofuran, dioxane, ethyl acetate, N, N-dimethylformamide, and the like. To a hydroxymoyl chloride compound represented by the following Chemical Formula 3 and treated in a solvent such as methylene chloride, chloroform, tetrahydrofuran in the presence of a base such as triethylamine to obtain an intermediate of Chemical Formula 4, which is then represented by Chemical Formula 8 or Ring addition reaction with an alkene or alkyne compound of 9 may be used to obtain a compound of Formula 5 or 6.

Alternatively, by using chlorax from the 3- or 4-pyridine aldehyde oxime compound of formula (2) and reacting with organic solvents such as chloroform, methylene chloride, ethyl acetate and the like in one step in a two-step solvent in water in the reaction system to formula (5) The compound of 6 can also be obtained.

The obtained compound of formula (5) or (6) is reacted with alkyl iodide to obtain an alkyl pyridine salt of formula (7) as shown in Scheme 2, which can be recrystallized or concentrated to be used in the next step. Subsequently, the obtained compound of formula 7 may be reduced with sodium borohydride to synthesize a compound of formula 1, which is the target compound of the present invention.

Compounds of formula (1) in the free state can be converted to acid addition salts by conventional methods of adding a solution containing a stoichiometric amount of the appropriate acid. For example, a pharmaceutically acceptable acid addition salt of Formula 1-HX may be prepared by adding an organic or inorganic acid of Formula HX to a compound of Formula 1 as described in Scheme 2.

Pharmaceutically acceptable acids include hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, acetic acid, benzoic acid, sulfonic acid, fumaric acid, maleic acid, citric acid, lactic acid, tartaric acid, oxalic acid or similar pharmaceutically acceptable organic and inorganic acids. .

The present invention also provides a pharmaceutical composition characterized by comprising, in addition to the conventional carrier, the compound of formula 1 or a pharmaceutically acceptable acid addition salt thereof as the active ingredient.

The novel compounds can be administered orally or parenterally, such as intravenously or intramuscularly, in conventional manner.

Dosage may vary depending on the age, condition, weight and mode of administration of the patient. The daily dose of the active ingredient is usually from about 0.01 to 200 mg per kg of body weight.

The novel compounds can be used in conventional solid or liquid formulations, for example in the form of uncoated tablets or (thin-coated) tablets, capsules, powders, granules or solutions. It is prepared by conventional methods. To this end, the active ingredient can be processed with conventional pharmaceutical auxiliaries such as tablet binders, extenders, preservatives, tablet disintegrants, rheology modifiers, plasticizers, wetting agents, dispersants, emulsifiers, solvents, sustained-release agents, and / or antioxidants. .

The compounds of the present invention are also useful for the treatment of neurological diseases and impaired cognition resulting from abnormalities in cholinergic neurotransmission such as Alzheimer's disease. The novel compounds of the present invention are also useful for pain, glaucoma, psychosis, schizophrenia, anxiety, bipolar disorder (bipolar psychosis), bipolar psychosis, depression, sleep disorders, epilepsy, cerebral ischemia, bowel incontinence and gastrointestinal motility disorders.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

Example 1: Preparation of 3-pyridine-carboxime

Dissolve 16.1 g (0.15 mol) of 3-pyridine carboaldehyde in 140 ml of methanol / water (1: 1) mixed solvent, add 13.5 g (1.3 equivalents) of hydroxylamine hydrochloride and 10.3 g (0.65 equivalents) of sodium carbonate. The reaction was carried out at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove methanol, extracted three times with ethyl acetate, washed with saturated aqueous sodium carbonate solution, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and recrystallized from ethanol to give 14.6 g (yield 80%) of 3 Pyridine-carboxime compound was obtained.

Example 2 Preparation of 3-Pyridine Hydroxymoyl Chloride

After dissolving 7 g of 3-pyridine-carboxime in 100 ml of tetrahydrofuran, 8.42 g (1.1 equivalents) of N-chloro succinimide was added and reacted at 50 ° C. for 1 hour. After completion of the reaction, the mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and quantitatively obtained 11.4 g of 3-pyridine hydroxymoyl chloride.

Example 3 Preparation of (3-Pyridin-3-yl-isoxazol-5-yl) -methanol

500 mg (3.19 mmol) of 3-pyridine hydroxymoyl chloride was dissolved in 10 ml of methylene chloride, 372 μl (1.1 equiv) of propazyl alcohol and 445 μl (1.2 equiv) of triethylamine were added thereto and reacted at room temperature for 1 hour. After completion of the reaction, the mixture was extracted three times with chloroform, washed twice with saturated aqueous sodium carbonate solution and once with water, and then filtered and dried over sodium sulfate and concentrated under reduced pressure. The dried compound was separated and purified by silica gel column chromatography using chloroform / methanol (30: 1 to 10: 1 gradient) as eluent, to obtain (3-pyridin-3-yl-isoxazol-5-yl) -methanol. 342 mg (61% yield) was obtained.

¹ H NMR (CDCl ₃ ): δ 8.98 (s, 1H), 8.68 (d, J = 3.6, 1H), 8.18 (d, J = 7.9, 1H), 7.44 (dd, J = 4.9, 7.7, 1H) , 6.64 (s, 1 H), 4.87 (s, 2 H), 2.90 (brs, 1 H); MS (m / e): 176 (M &lt; + &gt;), 145, 117, 78.

Example 4 Preparation of [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -methanol

291.2 mg (1.653 mmol) of (3-pyridin-3-yl-isoxazol-5-yl) -methanol obtained in Example 3 were dissolved in 5 ml of acetone, and then 206 µl (3 equivalents) of methyl iodide at 0 ° C. ) And reacted for 3 hours. The residue obtained by filtration was dried to give pyridine iodide salt. The salt compound was dissolved in 15 ml of ethanol / water (2: 1), and 75 mg (1.2 equivalents) of sodium borohydride was added at 0 ° C. for 1 hour. After completion of the reaction, 0.5 ml of acetone was added thereto, concentrated under reduced pressure, the remaining acetone and ethanol were removed, water was added to the residue, followed by extraction with chloroform three times, washed with brine and water, filtered with sodium sulfate, and concentrated under reduced pressure. The concentrated compound was separated and purified by silica gel column chromatography using chloroform / methanol (10: 1 to 5: 1 gradient) as the eluent to obtain the title compound [3- (1-methyl-1,2,5,6-tetra). 183.2 mg (57% yield) of hydro-pyridin-3-yl) -isoxazol-5-yl] -methanol were obtained. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.29 (m, 1H), 6.25 (s, 1H), 4.61 (s, 2H), 3.31 (t, J = 2.0, 2H), 2.59 (t, J = 5.7, 2H ), 2.39 (s, 3 H), 2.37 (m, 2 H); MS (m / e): 194 (M &lt; + &gt;), 193, 163.

Example 5 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanol

1- (3-pyridin-3-yl-isoxazole from 135 mg (0.862 mmol) of 3-pyridine hydroxymoyl chloride and 135 μl (2 equivalents) of 3-butyne-2-ol by the method of Example 3 above. 72.5 mg (61% yield) of -5-day) -ethanol were obtained.

¹ H NMR (CDCl ₃ ): δ 8.96 (s, 1H), 8.66 (d, J = 3.6, 1H), 5.10 (q, J = 6.7, 1H), 3.28 (brs, 1H), 1.66 (d, J) = 6.6, 3H); MS (m / e): 190 (M &lt; + &gt;), 145, 78.

From the method of Example 4 from 610.6 mg (3.21 mmol) of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanol obtained above, 1- [3- (1-methyl-1, 386.9 mg (58% yield) of 2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanol were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.20 (m, 1H), 6.12 (s, 1H), 4.88 (q, J = 6.5, 1H), 3.40 (d, J = 16, 1H), 3.18 (d, J = 16, 1H), 2.66 (q, J = 5.6, 1H), 2.6-2.3 (m, 6H), 1.50 (d, J = 6.6, 3H); MS (m / e): 194 (M &lt; + &gt;), 193, 163.

Example 6: Preparation of 1-methyl-3- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

3- (5-methoxymethyl-isoxazol-3-yl)-from 250 mg (1.60 mmol) of 3-pyridine hydroxymoyl chloride and 270 μl (2 equivalents) of methyl propazyl ether by the method of Example 3 above. 189 mg (62% yield) of pyridine was obtained.

¹ H NMR (CDCl ₃ ): δ 9.0 (d, J = 1.4, 1H), 8.71 (dd, J = 1.4, 3.5, 1H), 8.18 (ddd, J = 1.8, 3.8, 8.0, 1H), 7.45 ( dd.J = 4.9, 8.0, 1H), 6.64 (s, 1H), 4.63 (2, 3H), 3.49 (s, 3H); MS (m / e): 190 (M &lt; + &gt;), 145, 117, 78.

1-methyl-3- (5-methoxymethyl-iso by the method of Example 4 from 174 mg (0.915 mmol) of 3- (5-methoxymethyl-isoxazol-3-yl) -pyridine obtained above 95.8 mg (68% yield) of oxazol-3-yl) -1,2,5,6-tetrahydro-pyridine were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

Yield: 68%; ¹ H NMR (CDCl ₃ ): δ 6.40 (m, 1H), 6.35 (s, 1H), 4.52 (s, 2H), 3.43 (s, 3H), 2.65 (t, J = 5.8, 2H), 2.6- 2.4 (5H), 2.08 (m, 2H); MS (m / e): 208 (M &lt; + &gt; -1), 163, 120.

Example 7: Preparation of 3-pyridin-3-yl-isoxazole-5-carboxylic acid methyl ester

In addition, the 3-pyridine-carboxime compound 200 of Example 1 was cooled to 0 ° C. with 219 ml (1.5 equivalents) of methyl propiolate in a 5 ml solution of chloroform and cooled to 0 ° C. and stirred vigorously at 0 ° C. A solution of mg (1.638 mmol) 5 ml of chloroform was slowly added over 15 minutes. After reacting overnight at room temperature, the same procedure as in Example 3 was carried out to obtain 3-pyridin-3-yl-isoxazole-5-carboxylic acid methyl ester of Example 6 at 129.1 mg (39% yield). Synthesized.

¹ H NMR (CDCl ₃ ): δ 9.24 (s, 1H), 8.73 (d, J = 4.9, 1H), 8.19 (ddd, J = 1.9, 3.8, 8.0, 1H), 7.44 (dd, J = 4.9. 7.7, 1 H), 7.31 (s, 1 H), 4.02 (s, 3 H); MS (m / e): 204 (M &lt; + &gt;), 145, 117.

Example 8 Preparation of 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) isoxazole-5-carboxylic acid methyl ester

3-pyridine-3-yl-isoxazole-5-carboxylic acid methyl from 400 mg (2.56 mmol) of 3-pyridine hydroxymoyl chloride and 455 μl (2 equiv) of methyl propiolate by the method of Example 3 above. 205.1 mg (39% yield) of ester were obtained.

From 284.8 mg (1.40 mmol) of 3-pyridin-3-yl-isoxazole-5-carboxylic acid methyl ester obtained above or obtained in Example 7, 3- (1-methyl-1, 61.8 mg (25% yield) of 2,5,6-tetrahydro-pyridin-3-yl) -isoxazole-5-carboxylic acid methyl ester were prepared and obtained in Example 4, [3- (1-methyl) -1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -methanol was also partially produced by the overreduction reaction. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

Yield: 25%; ¹ H NMR (CDCl ₃ ): δ 6.33 (m, 1H), 6.27 (s, 1H), 4.58 (s, 3H), 4.58 (s, 3H), 3.29 (s, 2H), 2.56 (t, 2H, J = 6 Hz), 2.4-2.25 (m, 5H); MS (m / e): 193 (M &lt; + &gt;), 163, 152, 120.

Example 9 Preparation of 1-Methyl-3- (5-phenyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

698.0 mg (49) of 3- (5-phenyl-isoxazol-3-yl) pyridine from 1.0 g (6.39 mmol) of 3-pyridine hydroxymoyl chloride and 1.40 g (2 equivalents) of phenyl acetylene by the method of Example 3 above % Yield).

¹ H NMR (CDCl ₃ ): δ 9.09-9.08 (m, 1H), 8.73-8.70 (m, 1H), 8.24-8.20 (m, 1H), 7.88-7.85 (m, 2H), 7.51-7.41 (m , 4H).

From 498.0 mg (2.22 mmol) of 3- (5-phenyl-isoxazol-3-yl) pyridine obtained above, 1-methyl-3- (5-phenyl-isoxazol-3-yl was obtained by the method of Example 4. 196.0 mg (45% yield) of) -1,2,5,6-tetrahydro-pyridine was prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 7.78-7.27 (m, 5H), 6.60 (s, 1H), 6.47-6.44 (m, 1H), 3.45-3.43 (m, 2H), 2.48 (s, 3H), 2.47-2.43 (m, 2 H), 1.59 (s, 2 H); MS (m / e): 240 (M &lt; + &gt;), 196, 168, 135, 105, 94, 77.

Example 10 Preparation of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone

1- (3-pyridin-3-yl-isoxazole from 500 mg (3.19 mmol) of 3-pyridine hydroxymoyl chloride and 500 μl (2 equivalents) of 3-butyne-2-one by the method of Example 3 above. 387.2 mg (64% yield) of -5-day) -ethanone was obtained.

¹ H NMR (CDCl ₃ ): δ 8.74 (s, J = 4.7, 1H), 8.17 (dd J = 1.6, 7.9, 1H), 7.45 (dd, J = 4.8, 7.8, 1H), 7.45 (dd, J = 4.8, 7.8, 1H), 7.24 (s, 1H), 2.68 (s, 3H); MS (m / e): 188 (M &lt; + &gt;), 145, 117, 78.

Example 11: Preparation of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone oxime

100 mg (0.531 mmol) of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone obtained in Example 10 was dissolved in 10 ml of a methanol / water (1: 1) mixed solvent. Then, 55.4 mg (1.5 equivalents) of hydroxylamine hydrochloride and 42.2 mg (0.75 equivalents) of sodium carbonate were added and reacted at room temperature for 3 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove methanol, extracted three times with benzene / ethyl acetate, washed with saturated aqueous sodium carbonate solution, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 99.0 mg (yield 92%) of 1-. (3-Pyridin-3-yl-isoxazol-5-yl) -ethanone oxime compound was obtained as a Z / E (ratio of 16:84).

¹ H NMR (CDCl ₃ ): δ 8.63-8.67 (2d, J = 1.86, 1H), 8.25 (m, 1H), 7.9-7.8 (m, 1H), 7.2-7.1 (m, 1H), 6.61 (s , 1H), 1.95-1.88 (2s, 3H) (E / Z mixture); MS (m / e): 201 (M &lt; + &gt;), 145, 117, 78.

Example 12 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone oxime

Prepared by the method of Example 4 from 84.0 mg (0.413 mmol) of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone oxime obtained in Example 11, followed by chloroform / methanol (10 : 1 to 5: 1 gradient) was used as the eluent to separate and purify by silica gel column chromatography, and the desired compound 3-[(1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl)- Two stereoisomers of isoxazol-5-yl] -ethanone oxime were obtained in 17.3 mg (20% yield) and 19.5 mg (23% yield), respectively. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

Isomer (20% yield): ¹ H NMR (CDCl ₃ ): δ 6.52 (s, 1H), 6.37 (m, 1H), 3.38 (d, 2H, J = 2Hz), 2.61 (t, 2H, J = 5.7 Hz), 2.5-2.3 (m, 5H), 2.16 (s, 3H); MS (m / e): 221 (M &lt; + &gt;), 220, 204, 188, 179, 152, 120.

Main isomer (23% yield): ¹ H NMR (CDCl ₃ ): δ 7.26 (s, 1 H), 6.44 (m, 1 H), 3.42 (d, 2H, J = 1.9 Hz), 2.63 (t, 2H, J = 5.6 Hz), 2.5-2.3 (m, 5H), 2.26 (s, 3H); MS (m / e): 221 (M &lt; + &gt;), 220, 204, 188, 179, 152, 120.

Example 13 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone O-methyl oxime >

Example 100 was obtained from 100 mg (0.531 mmol) of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone obtained in Example 10 and 66.5 mg (1.5 equivalents) of methoxylamine hydrochloride. By method, 115.7 mg of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone O-methyl oxime was quantitatively obtained in a Z / E (ratio of 31:69) mixture.

¹ H NMR (CDCl ₃ ): δ 9.07-9.04 (2s, 1H), 8.70 (m, 1H), 8.19 (m, 1H), 7.44 (m, 1H), 6.90 (1s, 1H), 4.09-4.07 ( 2s, 3H) (E / Z mix); MS (m / e): 217 (M &lt; + &gt;), 145, 117

From the method of Example 4 from 96.8 mg (0.446 mmol) of 1- (3-pyridin-3-yl-isoxazol-5-yl) -ethanone O-methyl oxime obtained above, 1- [3- (1 -Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone O-methyl oxime 54 mg (56% yield) was converted to the Z-, E-isomer Prepared as a mixture in a ratio of 23:77. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 7.27, 6.6 (2s, 1H), 6.5-6.3 (2m, 1H), 4.04-4.03 (2s, 3H), 3.34 (m, 2H), 2.66 (m, 2H), 2.6-2.4 (m, 5H), 2.30-2.21 (2s, 3H), (E / Z mixed); MS (m / e): 235 (M &lt; + &gt;), 204, 188, 163, 135, 120.

Example 14 Preparation of 4-Pyridine Hydroxymoyl Chloride

6.67 g (yield 88%) of 4-pyridine hydroxymoyl chloride was synthesized in two steps using 7 g (65.35 mmol) of 4-pyridine carboxaldehyde as a starting material by the method of Examples 1 and 2. .

Example 15 Preparation of [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -methanol

From 400 mg (2.55 mmol) of 4-pyridine hydroxymoyl chloride obtained in Example 14, 138.6 mg (31) of (3-pyridin-4-yl-isoxazol-5-yl) -methanol was obtained. % Yield).

¹ H NMR (CDCl ₃ ): δ 8.65 (d, 2H, J = 5.79), 7.68 (d, 2H, J = 5.46), 6.61 (s, 1H), 4,77 (1s, 2H), 2.59 (brs , 1H); MS (m / e): 194 (M &lt; + &gt;), 163, 135, 120, 94, 68, 53.

From 118 mg (0.67 mmol) of (3-pyridin-4-yl-isoxazol-5-yl) -methanol obtained above, by the method of Example 4, [3- (1-methyl-1,2,5, 76.7 mg (99% yield) of 6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -methanol were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.21 (s, 1H), 6.15 (s, 1H), 4.61 (s, 2H), 3.10 (d, J = 2.61, 2H), 2.65-2.58 (m, 4H), 2.37 (s, 3 H).

Example 16 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanol

213.0 mg of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanol from the method of Example 3 from 500 mg (3.19 mmol) of 4-pyridine hydroxymoyl chloride obtained in Example 14 (35% yield) was obtained.

¹ H NMR (CDCl ₃ ): δ 8.67-8.65 (m, 2H), 7.69-7.67 (m, 2H), 6.57 (s, 1H), 5.03 (q, J = 6.60, 1H), 1.6 (d, J = 6.16, 3H); MS (m / e): 190 (M &lt; + &gt;), 145, 78, 51, 194 (M &lt; + &gt;), 163, 135, 120, 94, 68, 53.

From 193 mg (1.01 mmol) of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanol obtained above, by the method of Example 4, 1- [3- (1-methyl-1, 108.2 mg (78% yield) of 2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanol were prepared. In the following, oxalic acid and 1: 1 acid addition salt were made under diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.22 (s, 1H), 6.21 (s, 1H), 4.86 (q, J = 6.60, 1H), 3.11-3.05 (m, 2H), 2.66-2.59 (m, 4H ), 2.36 (3H, s), 1.48 (d, J = 6.66, 3H).

Example 17 Preparation of 1-methyl-4- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

400 mg (2.55 mmol) of 4-pyridine hydroxymoyl chloride obtained in Example 14 was used to prepare 150.0 mg (31) of 4- (5-methoxymethyl-isoxazol-3-yl) -pyridine. % Yield).

¹ H NMR (CDCl ₃ ): δ 8.80-8.71 (m, 2H), 7.70-7.68 (m, 2H), 6.69 (s, 1H), 4.61 (s, 2H), 3,48 (s, 3H); MS (m / e): 190 (M &lt; + &gt;), 145, 78, 51.

From 153.9 mg (0.81 mmol) of 4- (5-methoxymethyl-isoxazol-3-yl) -pyridine obtained above, by the method of Example 4, 1-methyl-4- (5-methoxymethyl-iso 80.9 mg (53% yield) of oxazol-3-yl) -1,2,5,6-tetrahydro-pyridine were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.33 (s, 1H), 6.29-6.27 (m, 1H), 4.52 (s, 2H), 3.43 (s, 2H), 3.15-3.13 (m, 2H), 2.67 ( s, 3H), 2.42 (s, 3H), 1.94 (s, 2H).

Example 18 Preparation of 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-4-yl) isoxazole-5-carboxylic acid methyl ester

97.2 mg of 3-pyridin-4-yl-isoxazol-5-yl-carboxylic acid methyl ester by the method of Example 6 from 400 mg (2.55 mmol) of 4-pyridine hydroxymoyl chloride obtained in Example 14 (17% yield) was obtained.

¹ H NMR (CDCl ₃ ): δ 8.79-8.77 (m, 2H), 7.75-7.73 (m, 2H), 7.32 (s, 1H), 4.03 (s, 3H).

From 82 mg (0.40 mmol) of 3-pyridin-4-yl-isoxazol-5-yl-carboxylic acid methyl ester obtained above by the method of Example 4, 3- (1-methyl-1,2,5, 4.4 mg (7% yield) of 6-tetrahydro-pyridin-4-yl) isoxazole-5-carboxylic acid methyl ester were prepared and obtained in Example 15 [3- (1-methyl-1,2, 5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -methanol was also produced by the overreduction reaction. And oxalic acid and 1: 1 acid addition salt were made under diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.33 (s, 1H), 6.28 (s, 1H), 4.52 (s, 2H), 3.43 (s, 2H), 3.19-3.18 (m, 2H), 2.71 (s, 3H), 2.45 (s, 3H).

Example 19 Preparation of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanone

From 800 mg (5.10 mmol) of 4-pyridine hydroxymoyl chloride obtained in Example 14, 313.1 mg of 3-pyridin-4-yl-isoxazol-5-yl-carboxylic acid methyl ester was obtained. (33% yield) was obtained.

¹ H NMR (CDCl ₃ ): δ 8.71 (d, J = 5.49, 2H), 7.72 (d, J = 5.22, 2H), 7.27 (s, 1H), 2.64 (s, 3H); MS (m / e): 188 (M &lt; + &gt;), 145, 117, 78, 51.

Example 20 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone oxime

From 18.0 (0.786 mmol) of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanone obtained in Example 19, 143.8 mg (90% yield) of yl-isoxazol-5-yl) -ethanone oxime was obtained.

¹ H NMR (CDCl ₃ ): δ 8.60-8.58 (m, 2H), 7.74-7.68 (m, 2H), 6.86 (s, 1H), 2.27, 2.21 (2s, 3H) (E / Z); MS (m / e): 203 (M &lt; + &gt;), 145, 78, 51.

The method of Example 12 from 128 mg (0.630 mmol) of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanone oxime obtained above, was obtained in the manner of 1- [3- (1-methyl- 1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone oxime, 29.7 mg (25% yield), 51.6, respectively mg (44% yield) was prepared. And oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

Isomers (25% yield, stomach); ¹ H NMR (CDCl ₃ ): δ 7.3 (s, 1H), 6.34 (s, 1H), 3.3-3.05 (m, 2H), 2.8-2.5 (m, 4H), 2.26 (s, 3H), 2.22 ( s, 3H); MS (m / e): 221 (M &lt; + &gt;), 204, 188, 179, 152, 135, 120, 94.

Main isomer (43% yield, below); ¹ H NMR (CDCl ₃ ): δ 6.66 (s, 1H), 6.35-6.31 (m, 1H), 3.15-3.13 (m, 2H), 2.71-2.62 (m, 4H), 2.39 (s, 3H), 2.19 (s, 3 H); MS (m / e): 221 (M &lt; + &gt;), 204, 188, 179, 163, 146, 135, 120, 94.

Example 21 Preparation of 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone O-methyl oxime >

From the method of Example 13, from 109.5 mg (0.582 mmol) of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanone obtained in Example 19, 1- (3-pyridine-4- 1-isoxazol-5-yl) -ethanone O-methyl oxime 123.5 mg (98% yield) were obtained.

¹ H NMR (CDCl ₃ ): δ 8.75-8.73 (m, 2H), 7.73-7.71 (m, 2H), 6.91 (s, 1H), 4.09, 4.07 (2s, 3H), 2.37, 2.28 (2s, 3H ) (E / Z); MS (m / e): 217 (M &lt; + &gt;), 145, 117, 78, 51.

108 mg (0.497 mmol) of 1- (3-pyridin-4-yl-isoxazol-5-yl) -ethanone O-methyl oxime obtained above according to Example 13 by 1- [3- (1- Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone O-methyl oxime two stereo (E / Z) isomers 15.8 mg (16%) each Yield), 55.2 mg (55% yield). Then oxalic acid and 1: 1 acid addition salt were each made in diethyl ether solution.

Isomers (16% yield, stomach); ¹ H NMR (CDCl ₃ ): δ 7.13 (s, 1H), 6.37 (m, 1H), 4.04 (s, 2H), 3.15 (d, 2H, J = 2.76), 2.8-2.6 (m, 4H), 2.42 (s, 3 H), 2.31 (s, 3 H); MS (m / e): 235 (M &lt; + &gt;), 207, 193, 188, 163, 135, 120, 94, 68, 53.

Main isomer (55% yield, below); ¹ H NMR (CDCl ₃ ): δ 6.60 (s, 1H), 6.31 (m, 1H), 4.02 (s, 2H), 3.14 (s, 2H), 2.66 (m, 4H), 2.40 (s, 3H) , 2.20 (s, 3 H); MS (m / e): 235 (M &lt; + &gt;), 207, 204, 188, 176, 163, 135, 120, 94, 68, 53.

Example 22 Preparation of 1-Methyl-3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 1.1 ml (2 equivalents) of ethyl vinyl ether by the method of Example 3, and then using hexane / ethyl acetate (1: 1) as eluent, silica gel Separation and purification by column chromatography gave 899.5 mg (81.4% yield) of 3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl) -pyridine.

¹ H NMR (CDCl ₃ ): δ 8.85-7.36 (m, 4H), 5.74 (dd, J = 1.70, 6.63, 1H), 3.97-3.89 (m, 1H), 3.70-3.59 (m, 1H), 3.42 (dd, J = 6.66, 17.76, 1H), 3.29 (d, J = 17.10, 1H), 1.27-1.22 (m, 3H); MS (m / e): 192 (M &lt; + &gt;), 175, 147, 118, 104, 78, 51.

Prepared by the method of Example 4 from 648.4 mg (3.37 mmol) of 3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl) -pyridine obtained above, followed by chloroform / methanol (30 1-methyl-3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl)-was purified by silica gel column chromatography using a: 1-15: 1 gradient) as the eluent. 463.4 mg (76% yield) of methyl-1,2,5,6-tetrahydro-pyridine were prepared. And oxalic acid and 1: 1 acid addition salt were made under diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.09-6.06 (m, 1H), 5.55 (dd, J = 1.68, 6.57, 1H), 3.90-3.82 (m, 1H), 3.61-3.51 (m, 1H), 3.32 -3.30 (m, 2H), 3.14 (dd, J = 6.57, 20.01, 1H), 2.97 (dd, J = 1.59, 1.89, 1H), 2.56-2.51 (m, 2H), 2.41-2.37 (m, 5H ), 1.21-1.18 (m, 2H); MS (m / e): 209 (M &lt; + &gt;), 150, 122, 109, 94.

Example 23 Preparation of 1-Methyl-3- (5-butoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 1.5 ml (2 equivalents) of butyl vinyl ether by the method of Example 3, and then using hexane / ethyl acetate (2: 1) as eluent, silica gel Separation and purification by column chromatography gave 934.7 mg (74% yield) of 3- (5-butoxy-4,5-dihydro-isoxazol-3-yl) -pyridine.

¹ H NMR (CDCl ₃ ): δ 8.85-7.35 (m, 4H), 5.72 (ddd, J = 1.74, 6.66, 1H), 3.92-3.84 (m, 1H), 3.60-3.53 (m, 1H), 3.42 (dd, J = 6.69, 17.25, 1H), 3.22 (ddd, J = 1.71, 17.43, 1H), 1.61-1.54 (m, 2H), 1.46-1.33 (m, 2H), 0.91 (t, J = 4.87 , 3H); MS (m / e): 220 (M &lt; + &gt;), 165, 147, 136, 78.

Prepared by the method of Example 4 from 684.3 mg (3.11 mmol) of 3- (5-butoxy-4,5-dihydro-isoxazol-3-yl) -pyridine obtained above, followed by chloroform / methanol (30 1-methyl-3- (5-butoxy-4,5-dihydro-isoxazol-3-yl)-was purified by silica gel column chromatography using a: 1-15: 1 gradient) as the eluent. 488.8 mg (74% yield) of 1,2,5,6-tetrahydro-pyridine were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.09-6.06 (m, 1H), 5.53 (dd, J = 1.59, 6.54, 1H), 3.82-3.77 (m, 1H), 3.54-3.45 (m, 1H), 3.32 -3.30 (m, 2H), 3.13 (dd, J = 6.45, 17.04, 1H), 2.96 (dd, J = 1.62, 17.04, 1H), 2.59-2.58 (m, 2H), 2.41-2.37 (m, 5H ), 1.58-1.50 (m, 2H), 1.39-1.31 (m, 2H), 0.89 (t, J = 7.35, 3H); MS (m / e): 237 (M &lt; + &gt;), 136, 122, 109, 94, 57.

Example 24 Preparation of 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-ol

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 1.06 mL (2 equiv.) Of vinyl acetate by the method of Example 3, and then using hexane / ethyl acetate (1: 1) as eluent. Chromatographic separation and purification, 526.7 mg (44.4% yield) and 3-isoxazol-3-yl-acetic acid 3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl ester 262.9 mg (31.3% yield) of pyridine was obtained.

¹ H NMR (CDCl ₃ ): δ 8.86-7.39 (m, 4H), 6.87 (dd, J = 1.44, 6.93, 1H), 3.66 (dd, J = 6.90, 17.82, 1H), 3.38 (dd, J = 1.44, 16.71, 1 H), 2.10 (s, 3 H); MS (m / e): 146 (M + -70), 118, 91, 78, 63.

From 413.7 mg (2.01 mmol) of the acetic acid 3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl ester obtained above by the method of Example 4, it is followed by chloroform / methanol (10 Isolation and purification by silica gel column chromatography using: 1) as eluent gave 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-iso 274.8 mg (86% yield) of oxazole-5-ol were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.07-6.03 (m, 1H), 5.84 (d, J = 6.75, 1H), 3.38 (d, J = 15.93, 1H), 3.23-3.10 (m, 2H), 2.92 (d, J = 17.04, 1H), 2.62-2.60 (m, 2H), 2.53-2.40 (m, 5H); MS (m / e): 182 (M &lt; + &gt;), 163, 136, 110, 94, 53.

Example 25 Preparation of 1-Methyl- (3-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

Prepared by the method of Example 4 from 154.4 mg (1.06 mmol) of 3-isoxazol-3-yl-pyridine obtained in Example 24, and then chloroform / methanol (30: 1 to 15: 1) was used as the eluent. Was purified by silica gel column chromatography to give 40.7 mg (33% yield) of 5-isoxazol-3-yl-1-methyl-1,2,5,6-tetrahydro-pyridine. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

3-isoxazol-3-yl-pyridine; ¹ H NMR (CDCl ₃ ): δ 9.04-7.40 (m, 4H), 8.58 (d, J = 1.74, 1H), 6.73 (d, J = 1.74, 1H); MS (m / e): 146 (M &lt; + &gt;), 118, 91, 66, 53.

1-methyl- (3-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine; ¹ H NMR (CDCl ₃ ): δ 8.03 (d, J = 1.74, 1H), 6.43 (d, J = 1.74, 1H), 6.42-6.40 (m, 1H), 3.42-3.40 (m, 2H), 2.62 -2.58 (m, 2 H), 2.46-2.40 (m, 5H); MS (m / e): 164 (M &lt; + &gt;), 122, 94, 66, 53.

Example 26 Preparation of 1-Methyl-3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

Prepared from 636 mg (4.06 mmol) of 3-pyridine hydroxymoyl chloride and 921 μl (2 equiv) of allyl ethyl ether by the method of Example 3, followed by hexane / ethyl acetate (1: 1) followed by chloroform / methanol ( 50: 1) was purified by silica gel column chromatography using eluent to give 948.9 mg (82% yield of 3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl) -pyridine )

¹ H NMR (CDCl ₃ ): δ 8.82-7.34 (m, 4H), 5.00-4.90 (m, 1H), 3.68-3.56 (m, 4H), 3.45-3.24 (m, 2H), 1.21 (t, J = 9.3, 3H); MS (m / e): 206 (M &lt; + &gt;), 147, 119, 105, 78, 59.

Prepared by the method of Example 4 from 690.4 mg (3.35 mmol) of 3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl) -pyridine obtained above, followed by chloroform / methanol ( 30: 1 to 15: 1 gradient) was used as the eluent, and purified by silica gel column chromatography to obtain 1-methyl-3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl 513.9 mg (75% yield) of 1,2,5,6-tetrahydro-pyridine was prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.04-6.01 (m, 1H), 4.75-4.71 (m, 1H), 3.58-3.46 (m, 4H), 3.27 (s, 2H), 3.12-2.98 (m, 2H ), 2.53 (t, J = 5.11, 2H), 2.39-2.36 (m, 5H), 1.19 (t, J = 7.03, 3H); MS (m / e): 224 (M &lt; + &gt;), 180, 165, 122, 94.

Example 27 Preparation of [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methanol

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 789 μl (2 equivalents) of allyl alcohol by the method of Example 3, and after using ethyl acetate, chloroform / methanol (20: 1) was used as an eluent. Separation and purification by column chromatography gave 786.6 mg (76% yield) of 3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl-methanol.

¹ H NMR (CDCl ₃ ): δ 8.84-7.35 (m, 4H), 4.96-4.90 (m, 1H), 3.96-3.91 (m, 1H), 3.74-3.70 (m, 1H), 3.46-3.29 (m , 2H); MS (m / e): 178 (M &lt; + &gt;), 147, 119, 78, 51.

Prepared by the method of Example 4 from 526.3 mg (2.95 mmol) of 3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl-methanol obtained above, followed by chloroform / methanol (9: 1 to 6: 1 gradient) was used as the eluent to separate and purify by silica gel column chromatography, to obtain [3- (1-methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5- 507.6 mg (96% yield) of dihydro-isoxazol-5-yl] -methanol were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 5.98-5.95M, 1H, 4.65-4.62 (m, 1H), 3.72-3.67 (m, 1H), 3.50-3.47 (m, 1H), 2.52-2.32 (m, 7H); MS (m / e): 196 (M &lt; + &gt;), 165, 152, 94.

Example 28 of 1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -ethanol Manufacture

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 1.0 mL (2 equivalents) of 3-butene-2-ol by the method of Example 3, and after chloroform / methanol (20: 1) after ethyl acetate Separation and purification by silica gel column chromatography using the eluent gave 843.9 mg (76% yield) of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanol. .

¹ H NMR (CDCl ₃ ): δ 8.82-7.32 (m, 4H), 4.74-4.61 (m, 1H), 4.16-4.13 (2m, 1H), 3.48-3.19 (m, 2H), 1.32-1.22 (M , 3H) (diaste); MS (m / e): 192 (M &lt; + &gt;), 147, 120, 105, 78, 51.

Prepared by the method of Example 4 from 547.0 mg (2.85 mmol) of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanol obtained above, followed by chloroform / Separation and purification by silica gel column chromatography using methanol (15: 1 to 7: 1 gradient) as eluent, 1- [3- (1-methyl-1,2,5,6-tetrahydropyridin-3-yl 492.2 mg (91% yield) of 4,5-dihydro-isoxazol-5-yl] -ethanol were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 5.99-5.95 (m, 1H), 4.43-4.34 (m, 1H), 4.00-3.96, 3.67-3.59 (2m, 1H), 3.19 (s, 1H), 3.13-2.79 (m, 2H), 2.49-2.43 (m, 2H), 2.34-2.30 (m, 5H), 1.17-1.08 (m, 3H); MS (m / e): 210 (M &lt; + &gt;), 165, 94, 53.

Example 29 Preparation of 3- (pyridin-3-yl) -4,5-dihydro-isoxazole-5-carboxylic acid methyl ester

Silica gel column was prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 1.03 ml (2 equiv) of methyl acrylate by the method of Example 3, using hexane / ethyl acetate (1: 1) as eluent. Separation and purification by chromatography gave 862.1 mg (73% yield) of 3-pyridin-3-yl-4,5-dihydro-isoxazole-5-carboxylic acid methyl ester.

¹ H NMR (CDCl ₃ ): δ 8.84-7.36 (m, 4H), 5.24 (dd, J = 7.44, 10.8, 1H), 3.83 (s, 3H), 3.73 (m, 2H); MS (m / e): 206 (M &lt; + &gt;), 147, 119, 78, 51.

Example 27 obtained by the method of Example 4 from 547.0 mg (2.85 mmol) of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanol obtained above; 413.2 mg (82% yield) of [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methanol Obtained.

Example 30 Preparation of 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazole-5-carbonitrile>

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 757 μl (2 equiv) of acrylonitrile by the method of Example 3, followed by hexane / ethyl acetate (1: 1) followed by chloroform / methanol (50 : 1) was separated and purified by silica gel column chromatography using eluent to obtain 776.2 mg (78% yield) of 3-pyridin-3-yl-4,5-dihydro-isoxazole-5-carbonitrile.

¹ H NMR (CDCl ₃ ): δ 8.83-7.40 (m, 4H), 5.43 (dd, J = 6.90, 10.14, 1H), 3.87-3.73 (m, 2H); MS (m / e): 173 (M &lt; + &gt;), 142, 120, 116, 78, 65, 51.

From 840.8 mg (2.67 mmol) of 3-pyridin-3-yl-4,5-dihydro-isoxazole-5-carbonitrile obtained above by the method of Example 4, followed by chloroform / methanol (30: 1 To 10: 1 gradient) as an eluent to separate and purify by silica gel column chromatography, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-di 133.0 mg (28% yield) of hydro-isoxazole-5-carbonitrile were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.18 (s, 1H), 5.07 (dd, J = 7.49, 10.90, 1H), 3.7-3.2 (m, 4H), 3.2-2.3 (m, 5H); MS (m / e): 165 (M &lt; + &gt; -CN), 147, 137, 121, 109, 94, 70, 65, 53.

Example 31 Preparation of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanone

Prepared from 900 mg (5.75 mmol) of 3-pyridine hydroxymoyl chloride and 960 μl (2 equiv) of methyl vinyl ketone by the method of Example 3, followed by hexane / ethyl acetate (1: 1) followed by chloroform / methanol (50 : 1) was purified by silica gel column chromatography using eluent to obtain 378.5 mg (35) of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanone (35 % Yield).

¹ H NMR (CDCl ₃ ): δ 8.87-7.41 (m, 4H), 5.15-5.09 (m, 1H), 4.001-3.83 (m, 1H), 3.77-3.65 (m, 1H), 2.36 (s, 3H ); MS (m / e): 190 (M &lt; + &gt;), 147, 119, 105, 78, 59.

Prepared by the method of Example 4 from 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -ethanone obtained above, and using 1- [3 of Example 28. -(1-methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -ethanol was obtained.

Example 32 1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -pyrroli Preparation of Dyn-2-one>

Prepared from 2.23 g (12.82 mmol) of 3-pyridine hydroxymoyl chloride and 2.74 mL (2 equiv) of N-vinyl-2-pyrrolidinone by the method of Example 3, followed by chloroform / methanol (30: 1 to 15 1- (3-gradient) was used as eluent to separate and purify by silica gel column chromatography, then 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -pyrrolidine-2 2.16 g (73% yield) of -on was obtained.

1 H NMR (CDCl 3): δ 8.84-7.37 (m, 4H), 6.70 (dd, J = 3.63, 9.96, 1H), 3.24-3.16 (m, 1H), 2.46-2.41 (m, 2H), 2.09-1.81 (m, 2H); MS (m / e): 231 (M &lt; + &gt;), 173, 147, 78.

From the method of Example 4 from 1.349 g (5.83 mmol) of 1- (3-pyridin-3-yl-4,5-dihydro-isoxazol-5-yl) -pyrrolidin-2-one obtained above After preparation, the mixture was purified by silica gel column chromatography using chloroform / methanol (30: 1 to 15: 1 gradient) as the eluent, and then purified by 1- [3- (1-methyl-1,2,5,6-tetra 902.5 mg (73.4% yield) of hydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -pyrrolidin-2-one were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.46 (dd, J = 3.35, 9.80, 1H), 6.30-6.01 (m, 1H), 3.27-3.11 (m, 4H), 2.86-2.83 (m, 1H), 2.52 -2.30 (m, 9H), 1.95-1.92 (m, 2H); MS (m / e): 249 (M &lt; + &gt;), 189, 164, 121, 109, 94.

Example 33 Preparation of 1-Methyl-3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine

Prepared from 1.30 g (7.47 mmol) of 3-pyridine hydroxymoyl chloride and 2.262 mg (2 equiv) of N-vinylphenyl sulfide by the method of Example 3, followed by chloroform / methanol (30: 1 to 15: 1 gradient) Was purified by silica gel column chromatography using the product as an eluent to obtain 1.22 g (57% yield) of 3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl) -pyridine.

Prepared by the method of Example 4 from 1.0 g (1.70 mmol) of 3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl) -pyridine obtained above, followed by chloroform / methanol ( 30: 1 to 15: 1 gradient) was used as eluent to separate and purify by silica gel column chromatography, and then purified by 1-methyl-3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl 929 mg (64% yield) of) -1,2,5,6-tetrahydro-pyridine were prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 7.58-7.55 (m, 5H), 6.02-5.97 (m, 2H), 3.51-3.48 (m, 2H), 3.13-3.11 (q, 2H), 2.57-2.54 (m , 2H), 2.43 (s, 3H); MS (m / e): 274 (M &lt; + &gt;), 165, 135, 121, 96, 81, 65, 53.

Example 34 3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -3a, 4,6,6a-tetrahydro-furo [3,4-d] iso Preparation of Oxazole>

Prepared from 1.30 g (7.47 mmol) of 3-pyridine hydroxymoyl chloride and 927 mg (2 equiv) of 2,4-dihydrofuran by the method of Example 3, followed by chloroform / methanol (30: 1 to 15: 1 Gradient), which was separated and purified by silica gel column chromatography using eluent to give 422.4 mg (27) of 3-pyridin-3-yl-3a, 4,6,6a-tetrahydro-furo [3,4-d] isoxazole % Yield).

¹ H NMR (CDCl ₃ ): δ 8.84-7.32 (m, 4H), 5.48-5.32 (m, 1H), 4.38-4.26 (m, 2H), 4.21-4.13 (m, 1H), 3.91-3.84 (m , 1H), 3.76-3.71 (m, 1H).

Prepared by the method of Example 4 from 322.4 mg (3.90 mmol) of 3-pyridin-3-yl-3a, 4,6,6a-tetrahydro-furo [3,4-d] isoxazole obtained above, Chloroform / methanol (30: 1 to 15: 1 gradient) was used as eluent to separate and purify by silica gel column chromatography, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl 194.2 mg (60% yield) of 3a, 4,6,6a-tetrahydro-puro [3,4-d] isoxazole was prepared. Then oxalic acid and 1: 1 acid addition salt were made in diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 5.99-5.96 (m, 1H), 5.25-5.21 (dd, J = 3.81, 9.27, 1H), 4.25-4.21 (d, J = 10.62, 1H), 4.20-4.10 ( dd, J = 1.4, 9.06, 1H), 4.03-3.97 (m, 1H), 3.81-3.70 (m, 2H), 3.28-3.23 (m, 2H), 2.59-2.34 (m, 7H); MS (m / e): 208 (M &lt; + &gt;), 179, 164, 149, 121, 106, 94, 53.

Example 35 1-Methyl-3- (5-methylsulfanylmethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine and dimethyl- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methyl-sulfonium iodide Manufacture

Prepared from 1.00 g (6.39 mmol) of 3-pyridine hydroxymoyl chloride and 861.6 mg (2 equiv) of allyl methyl sulfide by the method of Example 3, and then eluate chloroform / methanol (30: 1 to 15: 1 gradient). Separation and purification by silica gel column chromatography using to give 961.2 mg (72% yield) of 3- (5-methylsulfanylmethyl-4,5-dihydro-isoxazol-3-yl) -pyridine.

Chloroform / methanol (prepared from 861.2 mg (4.13 mmol) of 3- (5-methylsulfanylmethyl-4,5-dihydro-isoxazol-3-yl) -pyridine obtained by the method of Example 4 30: 1 to 15: 1) was purified by silica gel column chromatography using eluent to obtain 1-methyl-3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl). 259.4 mg (28% yield) of -1,2,5,6-tetrahydro-pyridine were prepared. And oxalic acid and 1: 1 acid addition salt were made under diethyl ether solution.

¹ H NMR (CDCl ₃ ): δ 6.10-6.00 (m, 1H), 4.84-4.75 (m, 1H), 3.25-3.15 (q, 2H), 3.04-2.97 (q, 2H), 2.81-2.75 (q , 2H), 2.65-2.40 (m, 4H), 2.39 (s, 3H), 2.18 (s, 3H); MS (m / e): 226 (M &lt; + &gt;), 179, 165, 137, 122, 94, 61, 53.

In this case, dimethyl- [3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl)-formed by binding methyl iodide to sulfide in reaction with methyl iodide. The 4,5-dihydro-isoxazol-5-yl-methyl-sulfonium iodide compound was separated and purified by silica gel column chromatography to obtain an additional 885.6 mg (58.3%) yield.

¹ H NMR (CDCl ₃ ): δ 6.39 (s, 1 H), 4.86-4.83 (m, 1 H), 3.51-3.48 (m, 2H), 3.32-3.29 (m, 4H), 3.17-3.11 (m, 6H ), 2.72-2.69 (m, 4H), 2.12 (s, 3H).

In order to demonstrate the usefulness of the target compound of formula 1 according to the present invention, the activity of the representative compounds as muscarinic acetylcholine receptor agonists was tested.

[Drug effect measurement]

The efficacy of the new anti-dementia candidates acting on the muscarinic receptor was first examined by affinity with the receptor through a muscarinic receptor affinity screening test, followed by [ ³ H] -oxotrimoline affinity screening test, guinea pig extraction. Organ functions using the ileum and phosphoinositide turnover measurements are used to characterize the compound's functionality (muscarin agonist / antagonist). Candidates selected from these are finally evaluated through the drug avoidance test (passive avoidance test) and general pharmacological / behavioral pharmacological measurement method for the memory and learning ability.

In vitro affinity screening test for agonist screening of muscarinic receptors

[ ³ H] N-methyl-scopolamine ([ ³ H] NMS) Affinity Search Test

To rule out possible interactions between subtypes, human recombinant mucarinic receptor subtypes 1, 2 and 3 expressed in CHO cells as receptor sources (M1, M2, M3, Biosignal, Canada). ) Was purchased and used. [ ³ H] N-methyl-scopolamine (NEN, NET-636) was used as the ligand attached to the radioisotope, and atropine was used for nonspecific binding force measurement. As the experimental instruments, liquid scintillation counter (MicroBeta 1450 plus), Inotech cell harvester (Inotech harvester, 96-well) and about 10 species were used.

To determine the effect of each compound on the M1 and M2 receptors, the ligands attached to the radioisotope were reacted with the receptors, and the remaining unbound ligands were filtered off with a glass fiber filter, followed by a washed filter disc. The amount of isotope remaining in the was determined to quantify the binding reaction of the ligand to the receptor and used to determine the affinity of the drug for the receptor.

The receptor, frozen at -70 ° C, was suspended in test buffer, followed by screening test (BioRad protein) to determine the optimal concentration of protein. 50 μl of hot-ligand and 10 μl of test drug were then added to a 96 well plate and 100 μl of 50 mM Tris buffer (pH 7.2) containing 10 mM MgCl ₂ and 1 mM EDTA as test buffer. Was added. The final volume of the reaction was 0.25 ml, the reaction was started by addition of 100 μl of diluted receptor suspension and allowed to react for 60 minutes on a 27 ° C. shake incubator. The test of the test drug was repeated four times and after 60 minutes the reaction was terminated by addition of cold Tris buffer (0.5 mL, pH 7.4). The reaction mixture was separated and washed with excess isotopes that did not bind to the receptor using an Inotech cell harvester system, and then radioactivity captured on the filter mat was measured by a liquid scintillation counter. One-step drug screening detects the affinity of the drug for the receptors for both concentrations (1 μM, 10 μM), and measures the effects at lower concentrations as a second step for drugs that showed significant effects. IC ₅₀ values were calculated by concentration gradient. The test drug was dissolved in dimethyl sulfoxide (DMSO) and diluted sequentially to the required concentration. The final concentration of 1% of the dimethyl sulfoxide reaction did not affect the binding. Kd and Bmax values from the saturation affinity screening test, and IC ₅₀ or Ki values of each test drug were calculated by observing nonlinear decline using a prism (Graphpad softwere Inc., USA).

Equilibrium affinity search test, ^[3 H] N- methyl-pole Scoring was measured using a step gradient of 12 amine, competition search test, 1 nM of ^[3 H] N- methyl-Scopolamine and 10 step gradient The binding force (IC ₅₀ ) was measured with a standard drug of. Standard drugs include 4-DAMP-methyl iodide, Pirenzepine, para-F-hexahydro-sila-defenidol, Metoctramine, etc. Used.

[ ³ H] -oxotrimorin-M ([ ³ H] Oxo-M) affinity search test

Experimental principle is the same as the ^[3 H] -NMS affinity search test, this experiment to confirm that the ^[3 H] (whether agonists or antagonists) Pharmacological functions of the compounds showing a high affinity from the affinities -NMS search test in the practice, ^[3 H] -NMS and ^[3 H] -Oxo-M (oxo-tree Maureen) affinity embodiment the choice test at the same time to measure the respective IC ₅₀ values, and when the ratio of the IC ₅₀ values obtained (NMS / The function can be known by obtaining Oxo-M). Generally, when the ratio is 180 or more, it is evaluated as a muscarinic receptor full agonist, 14 to 130 as a partial agonist, and 0.2 to 1.9 as an antagonist.

Preparation of Brain Membrane

Synaptic membrane required for Oxo-M binding receptor binding experiments of muscarinic receptors were prepared and used directly in the laboratory as follows. Sparague-Dawley Rat (Supplied from Laboratory Laboratory, Korea Research Institute of Chemical Technology, 250-300 g) was single-headed and immediately separated from brain tissue to obtain a forebrain. 10-fold ice-cold sucrose solution (0.32 M) was added followed by homogenization (10 strokes, 500 rpm) using a Teflon-glass homogenizer (Contorque, Eberbach) and 1000 g for 15 minutes. After centrifugation, the supernatant was again centrifuged at 17000 g for 20 minutes, and the pellet was stored and used at -20 ° C.

Before experimenting the above pellets, they were suspended in 10 ml of 20 mM HEPES buffer (pH 7.4) and then washed again by centrifugation at 17000 g for 15 minutes, and the last washed membrane was final concentration 1: 100 (wet wt / v). This was diluted in ice-cold 20 mM HEPES buffer (pH 7.4) and used for the experiment. Samples of all tests were repeated four times and 20 mM HEPES buffer (pH 7.4) was used as search test buffer. The final volume of the reaction was 1 ml and a 24-well plate (costar 24well) was used. 50 μl of hot-ligand (Oxo-M 5 nM) and 10 μl of test drug were included. The reaction was started by adding 750 μl of synaptic membrane and shaken at 30 ° C. for 40 minutes. In all experiments, non-specific binding was performed using 2 μM atropine, and after the reaction, a Wellac GF / C filter (WallacGF / C filter, 0.05% polyethyleneimine) was obtained using an Innotech cell harvester (24-channels, Inotech, Switzerland). Pre-soaking) under reduced pressure filtration [10x ice-cold saline] to separate the ligand bound to the receptor and the ligand that did not bind. The ligand bound to the receptor was measured for radioactivity by a liquid scintillation counter (MicroBeta Plus, Wallac, Finland).

The test drug was dissolved in dimethylsulfoxide (DMSO) and diluted sequentially to the required concentration. The final concentration of 1% of DMSO did not affect binding. Kd and Bmax values from the saturation affinity screening test, and IC ₅₀ or Ki values of each test drug were calculated by observing nonlinear decline using Prism (Graphpad softwere Icn., USA). Saturation affinity screening tests were conducted in the 0.1-10 nM [ ³ H] Oxo-M ligand range as described above.

Table 1 shows the binding of [ ³ H] -N-methyl scopolamine ([ ³ H] NMS) and [ ³ H] -oxotrimorine ([ ³ H] Oxo-M) to the mouse cortical receptors. The affinity for the muscarinic receptors of the representative compounds of the invention measured is illustrated.

Affinity of Representative Compounds for Muscarinic Receptors compound % Suppression NMS (10 μM) Oxo-M (1 μM) Example 12 21.4 - Example 24 0 46.0 Example 25 0 50.4 Example 26 17.1 33.3 Example 32 63.4 96.3 Example 33 24.6 33.2 Example 35 6.0 58.8

As can be seen from Table 1 above, the novel compounds according to the invention, in particular the compounds of Example 32, showed a high affinity for muscarinic receptors. In addition, the compounds are more potent and effective as partial agonists (agonists) (inhibition of the compound against N-methylscopolamine (NMS) and oxotrimorin (Oxo-M) Determined by rain).

The 1,2,5,6-tetrahydropyridine compound substituted with the isooxazole or isoxazoline derivative represented by the new formula (1) according to the present invention shows a good affinity for the muscarinic acetylcholine receptor, It is effective as a therapeutic agent for neurological diseases such as Alzheimer's disease.

Claims (8)

Substituted 1,2,5,6-tetrahydropyridine compounds of Formula 1 and their pharmaceutically acceptable salts. <Formula 1> In the above formula, R ² represents C _1-4 -alkyl, One of R ³ and R ⁴ , The other is H, R ⁵ is hydrogen, hydroxy, hydroxy-C _1-6 -alkyl, C _{1-6 -alkoxymethyl} , C _1-6 -alkoxy, acetoxy, C _1-4 -alkyl ester, nitrile, aryl, phenylthio , Methylthiomethyl, phenylsulfoxide, dimethylthiomethyl, C _1-4 -alkylketo, C _1-4 -alkylketooxime, C _1-4 -alkylketo-C _1-5 -alkyloxime, N-pyrroli Din-2-one, R ⁶ is hydrogen, R ⁵ and R ⁶ together form —CH ₂ OCH ₂ —. The method of claim 1, [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -methanol, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanol, 1-methyl-3- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) isoxazole-5-carboxylic acid methyl ester, 1-methyl-3- (5-phenyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone oxime, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -isoxazol-5-yl] -ethanone O-methyl oxime, [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -methanol, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanol, 1-methyl-4- (5-methoxymethyl-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-4-yl) isoxazole-5-carboxylic acid methyl ester, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone oxime, 1- [3- (1-Methyl-1,2,5,6-tetrahydro-pyridin-4-yl) -isoxazol-5-yl] -ethanone O-methyl oxime, 1-methyl-3- (5-ethoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 1-methyl-3- (5-butoxy-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-ol, 1-methyl-3- (isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 1-methyl-3- (5-ethoxymethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methanol, 1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -ethanol, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazole-5-carbonitrile, 1- [3- (1-Methyl-1,2,5,6-tetrahydropyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -pyrrolidin-2-one , 1-methyl-3- (5-phenylsulfanyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, 3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -3a, 4,6,6a-tetrahydro-puro [3,4-d] isoxazole, 1-methyl-3- (5-methylsulfanylmethyl-4,5-dihydro-isoxazol-3-yl) -1,2,5,6-tetrahydro-pyridine, or Dimethyl- [3- (1-methyl-1,2,5,6-tetrahydro-pyridin-3-yl) -4,5-dihydro-isoxazol-5-yl] -methyl-sulfonium iodide Droid, And pharmaceutically acceptable salts thereof. To obtain a nitrile oxide of the formula (4) by using the 3- or 4-pyridine aldehyde oxime compound of the formula (2) as a starting material, and by the ring addition reaction with the compound of the formula (8) or (9) to prepare a compound of the formula And reacting the compound of Formula 5 or 6 with C _1-4 -alkyl iodide to prepare a pyridine salt of Formula 7, followed by reduction reaction. <Formula 2> <Formula 4> <Formula 5> <Formula 6> <Formula 7> <Formula 8> <Formula 9> <Formula 1> Wherein R ¹ represents 3- or 4-pyridine and R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined in claim 1. The method of claim 3, wherein the nitrile oxide of Formula 4 is obtained by reacting the 3- or 4-pyridine aldehyde oxime compound of Formula 2 with chlorax. According to claim 3, wherein the nitrile oxide of formula 4 is obtained by reacting the 3- or 4-pyridine aldehyde oxime compound of formula (2) with N-chloro succinimide via a hydroxymoyl chloride compound of formula (3) Way to be. <Formula 3> Wherein R ¹ is as defined in claim 3. delete Alzheimer's disease, impaired cognitive function and oral dryness, comprising, as an active ingredient, a compound of formula 1 according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier Pharmaceutical composition for treatment. delete