EA045865B1 - SUBSTITUTED PIPERIDINE COMPOUNDS AND THEIR APPLICATION - Google Patents

Description

Technical field

The present invention relates to substituted piperidine compounds having orexin type 2 receptor activating activity, and to pharmaceutically acceptable salts thereof, as well as to medical use. Moreover, the present invention relates to medicinal products containing the compounds as active ingredients.

State of the art

Orexin-A (OX-A) and orexin-B (OX-B), two types of intracerebral neuropeptides specifically produced by specific neurons located in the outer field of the hypothalamus of the brain, have been discovered as endogenous ligands of orexin receptors (PTL 1-4), which are G protein coupled receptors and are present primarily in the brain (see PTL 5 and NPL 1). There are two known subtypes of orexin receptors: OX ₁ 1 subtype receptors (OX1R) and OX ₂ 2 subtype receptors (OX2R). Orexins have been found to promote feeding behavior in rats (NPL 1).

Mutation of the OX2R gene has been reported to be one of the causes of narcolepsy in dogs (NPL 2), and orexin knockout mice exhibit narcolepsy-like symptoms very similar to narcolepsy in humans or dogs (NPL 3). Studies using transgenic mice with modified orexin neurons and doubly transgenic mice generated by crossing these mice with transgenic mice overexpressing orexin have shown that the narcolepsy-like symptoms seen with modification of orexin neurons are eliminated by continuous expression of orexin (NPL 4). Similarly, intracerebroventricular administration of OX-A to transgenic mice with modified orexin neurons was found to suppress cataplexy-like inhibitory effects, and improve symptoms of narcolepsy, for example, by increasing alertness (NPL 4). Studies in OX2R knockout mice also suggest that OX2R is important in maintaining alertness (NPL 5). Damage to the orexin nerves has also been suggested to be a cause of daytime sleepiness in patients with Parkinson's disease (NPL 6). In addition, it has been suggested that plasma OX-A concentration levels in patients with sleep apnea are low (NPL 7). Based on this, it has been suggested that OX2R agonists may serve as treatments for narcolepsy or therapeutic agents for other sleep disorders in which hypersomnia is observed (NPL 8).

Therefore, it is believed that compounds with OX2R activating activity can be used as therapeutic agents for the treatment of narcolepsy, idiopathic hypersomnia, hypersomnia, sleep apnea syndrome, states of impaired consciousness such as coma, narcolepsy syndrome accompanied by narcolepsy-like symptoms, and hypersomnia syndrome. accompanied by daytime hypersomnia (for example, Parkinson's disease, Guillain-Barré syndrome and Kleine-Levin syndrome).

TAK-925, a compound with OX2R-activating activity, has entered phase I trials (intravenous infusion) in healthy subjects and patients with narcolepsy.

List of cited literature Patent literature sources

PTL 1. Publication of international patent application No. WO 1996/34877.

PTL 2. Unexamined Publication of Japanese Patent Application HEI No. 10-327888.

PTL 3. Unexamined Publication of Japanese Patent Application HEI No. 10-327889.

PTL 4. Unexamined Publication of Japanese Patent Application HEI No. 11-178588.

PTL 5. Unexamined Publication of Japanese Patent Application HEI No. 10-229887.

Non-patent literature

NPL 1. Sakurai T. et al., Cell, 1998, 92, 573-585.

NPL 2. Lin L. et al., Cell, 1999, 98, 365-376.

NPL 3. Chemelli R.M. et al., Cell, 1999, 98, 437-451.

NPL 4. Mieda M. et al., Proc. Natl. Acad. Sci. USA, 2004, 101, 4649-4654.

NPL 5. Willie J.T. et al., Neuron, 2003, 38, 715-730.

NPL 6. Thannickal T.C. et al., Brain. 2007, 130, 1586-1595.

NPL 7. Busquets X. et al., Respiration, 2004, 71, 575-579.

NPL 8. Mieda M. et al., CNS Drugs, 2013, 27, 83-90.

Short description

An object of the present invention is to provide piperidine compounds having orexin type 2 receptor activating activity, and pharmaceutically acceptable salts thereof, as well as pharmaceutical compositions containing them.

In particular, the present invention relates to the following embodiments.

The first embodiment provides a compound selected from the group consisting of: (2R)-2-cyclopropyl-2-{(1R,3S,5S)-3-[(3S,4R)-1-(5-fluoropyrimidine-2 -yl)-3-methoxypiperidin-4yl]-8-azabicyclo[3.2.1]octan-8-yl}acetamide represented by the following formula (I):

- 1 045865

^)-2-((^^^)-3-(^.^)-1-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octane -8-yl)-3-methylbutanamide represented by the following formula (II):

^)-2-((^^^)-3-(^.^)-1-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8-azabicyclo-

[3.2.1]oc tan-8-yl)-2-cyclopropylacetamide represented by the following formula (III):

(R)-2-cyclopropyl-2-((1R,3S,5S)-3-((2S,4S)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8azabicyclo [3.2.1]octan-8-yl)acetamide represented by the following formula (IV):

or a pharmaceutically acceptable salt thereof.

The second embodiment provides (2R)-2-cyclopropyl-2-{(1R,3S,5S)-3-[(3S,4R)-1-(5fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl ]-8-azabicyclo[3.2.1 ]octan-8-yl}acetamide represented by the following formula (I):

or a pharmaceutically acceptable salt thereof.

In a third embodiment, (R)-2-((1R,3S,5S)-3-((3S,4R)-1-(5-fluoropyrimidin2-yl)-3-methoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-3-methylbutanamide represented by the following formula (II):

or a pharmaceutically acceptable salt thereof.

In a fourth embodiment, (R)-2-((1R,3S,5S)-3-((3S,4R)-1-(5chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8- azabicyclo[3.2.1]octan-8-yl)-2-cyclopropylacetamide represented by the following formula (III):

or a pharmaceutically acceptable salt thereof.

The fifth embodiment provides ^)-2-cyclopropyl-2-(( 1 R,3S,5S)-3-((2S,4S)-1 -(5fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl )-8-azabicyclo[3.2.1 ]octan-8-yl)acetamide represented by the following formula (IV):

- 2 045865

△ or a pharmaceutically acceptable salt thereof.

In a sixth embodiment, a pharmaceutical composition is provided containing a compound or a pharmaceutically acceptable salt thereof according to any one of options 1-5 above.

In a seventh embodiment, an orexin type 2 receptor agonist is provided, comprising a compound or a pharmaceutically acceptable salt thereof of any one of embodiments 1-5 above.

In an eighth embodiment, a therapeutic agent for treating narcolepsy is provided, comprising a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above.

In a ninth embodiment, a method of treating narcolepsy is provided, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1-5 above.

In a tenth embodiment, a method of activating an orexin receptor type 2 is provided, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1-5 above.

In an eleventh embodiment, a method of treating narcolepsy is provided, comprising administering to a subject a compound or a pharmaceutically acceptable salt thereof according to any one of options 1-5 above.

The twelfth embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above for the manufacture of a pharmaceutical composition for the treatment of narcolepsy.

A thirteenth embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof of any one of embodiments 1-5 above for the treatment of narcolepsy.

In a fourteenth embodiment, a therapeutic agent for treating cataplexy is provided, comprising a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above.

In a fifteenth embodiment, a method of treating cataplexy is provided, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of options 1-5 above.

A sixteenth embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof of any one of embodiments 1-5 above for the treatment of cataplexy.

A seventeenth embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above for the manufacture of a pharmaceutical composition for the treatment of cataplexy.

An eighteenth embodiment provides a therapeutic agent for treating hypersomnia syndrome comprising a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above.

In a nineteenth embodiment, a method of treating hypersomnia syndrome is provided, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1-5 above.

The twentieth embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof of any one of embodiments 1 to 5 above for the treatment of hypersomnia syndrome.

The twenty-first embodiment provides the use of a compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5 above for the manufacture of a pharmaceutical composition for treating hypersomnia syndrome.

Substituted piperidine compounds represented by formulas (I), (II), (III) and (IV) (hereinafter referred to as compounds (I), (II), (III) and (IV)), or pharmaceutically acceptable salts thereof, according to the present invention have orexin receptor type 2 activating activity, as demonstrated by the activity data in the example pharmacological tests described below. Compounds (I), (II), (III) and (IV) of the present invention have orexin receptor type 2 activating activity and can be used as therapeutic agents for the treatment of narcolepsy.

Brief description of graphic materials

In fig. 1 is an ORTEP diagram showing the results of X-ray crystallographic analysis of the compound prepared in Example 1.

In fig. 2 is an ORTEP diagram showing the results of X-ray crystallographic analysis of the compound prepared in Example 3 (a dimer containing

- 3 045865 two added molecules of acetonitrile).

In fig. 3 is an ORTEP diagram showing the results of X-ray crystallographic analysis of the compound obtained in Example 4.

Detailed description

Next, the present invention will be explained in detail.

Polymorphic crystals of the compounds of the present invention may also exist, and any crystalline form or mixture thereof, as well as amorphous forms, can be used without any limitation, and the compounds of the present invention also include both anhydrous and solvated (especially hydrated) forms.

Isotopically labeled forms of compounds (I), (II), (III) and (IV) are also covered by the present invention. An isotopically labeled compound is the same as any of compounds (I), (II), (III) and (IV), except that one or more atoms are replaced by an atom having a different atomic mass or mass number than the atomic mass mass or mass number commonly found in nature. Examples of isotopes that may be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, phosphorus, sulfur, iodine and chlorine, in particular ^2H , ^3H , C, ^14C , ^15N , ¹⁸ O, ¹⁸ F, ³² P, ³⁵ S, ¹²³ I and ¹²⁵ I.

Isotopically labeled compounds, such as those containing radioactive isotopes such as ³ H and/or ¹⁴ C, are useful for tissue distribution assays of drugs and/or substrates. Isotopes ³ H and ¹⁴ C are considered suitable due to the ease of their preparation and detection. The ¹¹ C and ¹⁸ F isotopes are considered suitable for PET (positron emission tomography), while the ¹²⁵ I isotope is considered suitable for SPECT (single photon emission computed tomography); and all are suitable for brain imaging. Substitution of ^2H and similar isotopes with heavier isotopes provides advantages in certain treatments, such as longer half-life in vivo and lower dosages required due to higher metabolic stability, and are therefore considered useful in some circumstances. Such isotope-labeled compounds can be prepared in a similar manner using the procedures disclosed in the examples below, using readily applicable isotope-labeled reagents instead of non-isotope-labeled reagents.

The pharmaceutically acceptable salt mentioned herein is not particularly limited as long as it is formed with the compound of the present invention, and specific examples include acid addition salts such as inorganic acid salts, organic acid salts or acidic amino acid salts.

Unless otherwise specified, the term pharmaceutically acceptable salt as used herein refers to one in which the number of acid molecules per molecule of the compound is, without limitation, preferably from about 0.5 to about 2 acid molecules per molecule compound or more preferably about 0.5, about 1 or about 2 molecules of acid relative to one molecule of salt compound formed in the appropriate proportion.

Preferable examples of inorganic acid salts include hydrochlorides, hydrobromides, sulfates, nitrates and phosphates, and preferred examples of organic acid salts include acetates, succinates, fumarates, maleates, tartrates, citrates, lactates, stearates, benzoates, methanesulfonates, n-toluenesulfonates and benzenesulfonates.

Preferable examples of acidic amino acid salts include aspartic acid salts and glutamic acid salts.

When any of the compounds (I), (II), (III) and (IV) of the present invention are obtained in free form, it can be converted into a suitable salt or hydrate of compound (I), (II), (III) or ( IV) in the usual way.

When preparing any of the compounds (I), (II), (III) and (IV) of the present invention in the form of a salt of compound (I), (II), (III) or (IV) or a hydrate of compound (I), ( II), (III) or (IV) it can be converted into the free form of compound (I), (II), (III) or (IV) in the usual way.

The various isomers (e.g., optical isomers, rotational isomers and stereoisomers) obtained for compounds (I), (II), (III) and (IV) of the present invention can be purified and isolated using conventional separation means, such as, for example, recrystallization, diastereomer salt method, enzymatic separation or chromatographic methods (eg thin layer chromatography, column chromatography or gas chromatography).

Compound

The pharmaceutical composition of the present invention can be prepared by mixing a pharmaceutically acceptable additive with a compound selected from the group of compounds (I), (II), (III) and (IV), or a pharmaceutically acceptable salt thereof. The pharmaceutical composition of the present invention can be prepared by a known method, such as the method described in the General Preparation Rules section of the Japanese Pharmacopoeia, 17th edition.

The pharmaceutical composition of the present invention can be suitably

- 4 045865 was administered to the patient in accordance with the dosage form.

The dosage of any of the compounds (I), (II), (III) and (IV) or their pharmaceutically acceptable salts of the present invention may vary depending on the severity of symptoms, age, sex and body weight of the patient, dosage form or type of salt and the specific type of disease, but generally it is from about 30 µg to 10 g, preferably from 100 µg to 5 g and even more preferably from 100 µg to 1 g for oral administration and from about 30 µg to 1 g, preferably from 100 µg to 500 mg and even more preferably 100 µg to 300 mg for administration by injection per day for an adult in a single dose or in divided doses.

As used herein, an orexin receptor type 2 agonist refers to an agent that binds to the orexin receptor type 2 and has the effect of activating the orexin receptor type 2, as well as a ligand in vivo.

The compounds of the present invention can also be used as chemical probes to capture target proteins of physiologically active small molecule compounds. In particular, the compounds of the present invention can be converted into affinity chromatography probes or photoaffinity probes by introducing tag groups or linkers into those portions of the compounds other than the structural portions necessary for their activity, using the method described in J. Mass Spectrum. Soc. Jpn. Vol. 51, No. 5, 2003, p. 492-498 or, for example, in WO 2007/139149.

The tag group or linker used in the chemical probe may be a group from, for example, any of the following (1) to (5):

(1) protein tag groups, including photoaffinity tag groups (e.g., benzoyl, benzophenone, azide, carbonyl azide, diaziridine, enone, diazo and nitro groups) and chemical affinity groups (e.g., ketone groups with an alpha carbon atom substituted by halogen, carbamoyl, ester and alkylthio groups, α,β-unsaturated ketones, Michael acceptors such as esters and oxirane);

(2) cleavable linkers such as -S-S-, -O-Si-O-, monosaccharides (such as glucose or galactose) and disaccharides (such as lactose) and oligopeptide linkers cleavable by enzymatic reactions;

(3) marker groups for fluorescence in situ hybridization, such as biotin, 3-(4,4-difluoro5,7-dimethyl-4H-3a and 4a-diaza-4-bora-indacen-3-yl)propionyl;

(4) detectable markers, including radioactive tag groups such as ¹²⁵ I, ³² P, ³ H and ¹⁴ C; fluorescent tag groups such as fluorescein, rhodamine, dansyl, umbelliferone, 7-nitrofurazanil and 3-(4,4-difluoro-5,7-dimethyl-4H-3a and 4a-diaza-4-bora-indacen-3-yl )propionyl group; chemiluminescent groups such as lumiferin and luminol; and heavy metal ions such as lanthanide metal ions and radium ion; or (5) groups associated with solid phase supports such as glass beads, glass plates, microtiter plates, agarose beads, agarose layers, polystyrene beads, polystyrene layers, nylon beads and nylon layers.

The probe obtained by introducing a tag group selected from items (1) to (5) above into the compound of the present invention in the manner described in the above-mentioned literature can be used as a chemical probe to identify tagged proteins that are useful for discovery new innovative goals in drug development.

Examples

The compounds (I), (II), (III) and (IV) of the present invention can be prepared using the methods described in the following examples, and the effects of the compounds can be confirmed using the methods described in the following test examples. However, these specific examples are illustrative only and are not intended to limit the present invention in any way, and various modifications may be made without departing from the scope of the present invention.

The compounds mentioned by reference to published documents were prepared by the method described in these documents.

Abbreviations used throughout this document are well known to those skilled in the art. In particular, the following are used:

n.: normal;

tert-: tertiary;

1H NMR: proton nuclear magnetic resonance spectrometry;

MS: mass spectrometry;

HPLC: High Performance Liquid Chromatography.

The term room temperature as used in all examples and preparation examples generally refers to the range from about 10 to 35°C. Percentages are percentages by weight unless otherwise noted.

Chemical shifts in proton nuclear magnetic resonance spectra are recorded in a single

- 5 045865 δ (ppm) with respect to tetramethylsilane, and the interaction constants are recorded in hertz (Hz). The patterns are represented as s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad and brs: broad singlet.

Mass spectrometry was performed using Acquity UPLC™ or Acquity UPC ² ™ from Waters Co.

For chromatography, Silica Gel60 from Merck (70-230 mesh or 230-400 mesh ASTM) or PSQ60B from Fuji Silysia Chemical, Ltd. was used as silica gel. or used a prepackaged column {column: Yamazen Hi-Flash™ Column (silica gel), size: S (16x60 mm), M (20x75 mm), L (26x100 mm), 2L (26x150 mm) or 3L (46x130 mm) , or Biotage™ SNAP Ultra Silica Cartridge from Biotage Co., size: 10 g, 25 g or 50 g}. Supercritical fluid chromatography (SFC) fractionation was performed using Prep100q from Waters Co.

CHROMATOREX NH-DM2035 from Fuji Silysia Chemical Ltd. was used as NH silica gel. or used a prepackaged column {column: Yamazen Hi-Flash™ Column (Amino), size: S (16x60 mm), M (20x75 mm), L (26x100 mm), 2L (26x150 mm) or 3L (46x130 mm) , or Presep™ (Luer Lock) NH ₂ (HC) from Wako Pure Chemical Industries, Ltd., size: Type M (14 g/25 ml), Type L (34 g/70 ml), Type 2L (50 g/ 100 ml) or type 3L (110 g/200 ml)}.

The nomenclature of the following compounds follows that specified in E-Notebook version 12 or 13 (Perkin-Elmer), except for commonly used reagents.

Receipt example 1.

Synthesis of 2-bromo-2-cyclopropylacetamide

Oxalyl chloride (CAS No. 79-37-8) (3.11 ml, 36.3 mmol) and N,N-dimethylformamide (60.0 µl, 0.775 mmol) were added to the cyclopropylacetic acid solution (CAS No. 5239-82- 7) (3.30 g, 33.0 mmol) in 1,2-dichloroethane (60 ml) and the mixture was stirred for 40 min at room temperature. Hydrobromic acid (56.0 mg, 0.330 mmol) and N-bromosuccinimide (CAS No. 128-08-5) (7.04 g, 39.6 mmol) were added to the reaction mixture, which was then refluxed for 18 h. The reaction mixture was added to ammonia (28% aqueous solution, 60 ml, 2.77 mol) at 0°C, and then ethyl acetate and sodium hydroxide (2N aqueous solution) were added to separate. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting solid was triturated with ethyl acetate/n-heptane and the precipitate was filtered off. The resulting solid was dried under reduced pressure to obtain the title compound (1.20 g).

MS (ESI) mass/charge: 178 [M+H]+.

Receipt example 2.

Synthesis of 3-methoxyisonicotinonitrile

Sodium ethoxide (CAS No. 124-41-4) (3.90 g, 72.2 mmol) was added to a solution of 3-chloro-4cyanopyridine (CAS No. 68325-15-5) (5.00 g, 36.1 mmol) in tetrahydrofuran (36.0 ml) at room temperature and the mixture was refluxed for 1 hour. Water and ethyl acetate were added to the reaction mixture and the organic layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting solid was triturated with ethyl acetate/n-heptane and the precipitate was filtered off. The resulting solid was dried under reduced pressure to obtain the title compound (1.91 g).

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 4.06 (s, 3H), 7.44 (d, J=5.0 Hz, 1H), 8.37 (d, J=4, 5 Hz, 1H), 8.49 (s, 1H).

MS (ESI) mass/charge: 135 [M+H] ⁺ .

Receipt example 3.

Synthesis of (R)-2-6pom-3-methylbutanamide

Oxalyl chloride (9.28 ml, 108 mmol) and N-dimethylformamide (30.0 µl, 0.387 mmol) were added to a solution of (P)-2-bromo-3-methylbutyric acid (CAS No. 76792-22-8) (9 .80 g, 54.1 mmol) in methylene chloride (100 ml) at 0°C and the mixture was stirred for 6 hours at room temperature. The reaction mixture was added to ammonia (28% aqueous solution, 50.0 ml, 2.31 mol) at 0°C and then extracted with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting solid was triturated with

- 6 045865 ethyl acetate/n-heptane and the precipitate was filtered off. The resulting solid was dried under reduced pressure to obtain the title compound (8.20 g).

1H NMR (400 MHz, CDCl3) δ (ppm): 1.01 (d, J=6.3 Hz, 3H), 1.07 (d, J=6.8 Hz, 3H), 2.34-2 .39 (m, 1H), 4.28 (d, J=4.5 Hz, 1H), 5.58 (brs, 1H), 6.41 (brs, 1H).

MS (ESI) mass/charge: 180 [M+H]+.

Receipt example 4.

Synthesis of (1R,3s,5S)-3-((3S,4R)-1-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octane

(1) Synthesis of tert-butyl-(1R,3r,5S)-3-hydroxy-3-(3-methoxyniridin-4-yl)-8-azabicyclo[3.2.1]octane8-carboxylate.

After adding 3-methoxyisonicotinonitrile (1.19 g, 8.88 mmol) and bis-(pinacolato)diboron (CAS No. 73183-34-3) (2.25 g, 8.88 mmol) to the solution butoxycarbonyl)nortropinone (CAS No. 185099-67-6) (1.00 g, 4.44 mmol) in methyl tert-butyl ether (18.0 ml) the mixture was refluxed for 16 hours. Sodium carbonate ( 2 mol/L aqueous solution, 20.0 ml) was added to the reaction mixture at 0°C, and the mixture was stirred for 20 minutes at 0°C. Ethyl acetate was added to the reaction mixture, and the organic layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 0-20% methanol/ethyl acetate) to give the title compound (1.12 g).

1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.48 (s, 9H), 1.77 (m, 2H), 1.94-1.99 (m, 2H), 2.21-2 .32 (m, 2H), 2.37-2.48 (m, 1H), 2.63-2.74 (m, 1H), 2.78 (s, 1H), 3.96 (s, 3H ), 4.22-4.29 (m, 1H), 4.31-4.42 (m, 1H), 7.28 (d, J=5.4 Hz, 1H), 8.22-8, 25 (m, 2H).

MS (ESI) mass/charge: 335 [M+H]+.

(2) Synthesis of tert-butyl-(1R,5S)-3-(3-methoxypyridin-4-yl)-8-azabicyclo[3.2.1]oct-2-ene-8carboxylate.

Concentrated sulfuric acid (1.60 mL, 30.0 mmol) was added to tert-butyl-(1R,3r,5S)-3hydroxy-3-(3-methoxypyridin-4-yl)-8-azabicyclo[3.2.1] octane-8-carboxylate (610 mg, 1.82 mmol) and the mixture was stirred for 40 minutes at room temperature. The reaction mixture was added to a solution of potassium hydroxide (5.00 g, 89.1 mmol) in water (10.0 ml) at 0°C. Tetrahydrofuran (10.0 ml) and di-tert-butyl dicarbonate (CAS No. 24424-99-5) (478 mg, 2.19 mmol) were added to the reaction mixture, which was then stirred for 10 minutes at room temperature. Ethyl acetate was added to the reaction mixture, and the organic layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 5-100% ethyl acetate/n-heptane) to give the title compound (420 mg).

1H NMR (400 MHz, CDCl3) δ (ppm): 1.46 (s, 9H), 1.72-1.81 (m, 1H), 1.97-2.03 (m, 2H), 2. 05-2.36 (m, 2H), 2.94-3.26 (m, 1H), 3.87 (s, 3H), 4.21-4.61 (m, 2H), 6.30 ( brs, 1H), 6.99 (d, J=4.5 Hz, 1H), 8.17 (d, J=5.0 Hz, 1H), 8.21 (s, 1H).

MS (ESI) mass/charge: 317 [M+H]+.

(3) Synthesis of tert-butyl-(^^)-3-(3-methoxypyridin-4-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate.

After adding 10% palladium on carbon (AD, 52.7% aqueous, 284 mg, 0.126 mmol, product of Kawaken Fine Chemicals Co., Ltd.) to a solution of tert-butyl-(^^)-3-(3-methoxypyridine -4-yl)-8azabicyclo[3.2.1]oct-2-ene-8-carboxylate (400 mg, 1.26 mmol) in methanol (3.00 ml) the mixture was stirred for 1 h at room temperature under a hydrogen atmosphere . The reaction mixture was filtered through

- 7 045865

Celite™ and the residue were washed with ethyl acetate. The resulting filtrate was concentrated under reduced pressure to obtain a mixture of endo form and exo form (endo:exo = 1:2, 398 mg).

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.49 (s, 6H), 1.50 (s, 3H), 1.61-2.13 (m, 22/3H), 2. 40-2.45 (m, 2/3H), 2.96-3.01 (m, 1/3H), 3.47-3.59 (m, 2/3H), 3.88 (s, 1H ), 3.90 (s, 2H), 4.16-4.28 (m, 1H), 4.34 (brs, 1H), 7.04 (d, J=4.5 Hz, 2/3H) , 7.06 (d, J=5.0 Hz, 1/3H), 8.13-8.23 (m, 2H).

MS (ESI) mass/charge: 319 [M+H]+.

(4) Synthesis of tert-butyl-(^^^)-3-(3-methoxypyridin-4-yl)-8-azabicyclo[3.2.1]octane-8carboxylate.

Potassium tert-Butoxide (281 mg, 2.50 mmol) was added to a solution of tert-butyl (1K^)-3-(3methoxypyridin-4-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (398 mg, 1.25 mmol) in t-butanol (4.00 mL) and the mixture was refluxed for 17 hours. Ethyl acetate and brine were added to the reaction mixture and the organic layer was separated. Then, the organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to obtain the title compound (385 mg).

^1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.49 (s, 9H), 1.58-2.06 (m, 8H), 3.49-3.56 (m, 1H), 3.90 (s, 3H), 4.24 (brs, 1H), 4.34 (brs, 1H), 7.04 (d, J=5.0 Hz, 1H), 8.17-8.19 (m, 2H).

MS (ESI) mass/charge: 319 [M+H]+.

(5) Synthesis of tert-buty.g-(1 R,3s,5S)-3 -(1 -(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo [3.2.1 ] octane-8-carboxylate.

After adding 10% palladium on carbon (AD, 52.7% aqueous, 7.91 g, 3.52 mmol, product of Kawaken Fine Chemicals Co., Ltd.) to a solution of tert-butyl-(^^^)-3 -(3-methoxypyridin-4-yl)-8azabicyclo[3.2.1]octane-8-carboxylate (11.2 g, 35.2 mmol) in acetic acid (100 ml) the mixture was stirred for 18 h at 70°C in a hydrogen atmosphere. The reaction mixture was filtered through Celite™ and the residue was washed with ethyl acetate. The filtrate was concentrated under reduced pressure. Ethyl acetate and sodium hydroxide (2N) were added to the residue, and the organic layer was separated. The organic layer was dried with ISOLUTE™ HM-N, filtered and concentrated under reduced pressure. After adding KH-dimethylformamide (50.0 ml), 2-chloro-5-fluoropyrimidine (CAS No. 62802-42-0) (5.21 ml, 42.2 mmol) and potassium carbonate (7.29 g) to the residue , 52.8 mmol) the mixture was stirred for 40 min at 80°C. Ethyl acetate and brine were added to the reaction mixture, and the organic layer was separated. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, 10-60% ethyl acetate/n-heptane) to give the title compound (9.84 g).

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.19-1.30 (m, 3H), 1.46 (s, 9H), 1.46-1.65 (m, 6H), 1.81-1.97 (m, 3H), 2.68 (d, J=14.5 Hz, 1H), 2.71-2.81 (m, 1H), 3.30 (s, 3H) , 3.35 (brs, 1H), 4.08-4.31 (m, 2H), 4.64-4.77 (m, 1H), 5.04-5.18 (m, 1H), 8 .13(s,2H).

MS (ESI) mass/charge: 421 [M+H]+.

(6) Synthesis of tert-butyl-(1R,3s,5S)-3-((3S,4R)-1-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)8-azabicyclo[3.2 .1]octane-8-carboxylate.

The compound tert-butyl-(1 R.3s.5S)-3 -(1 -(5-fluoropyrimidin-2-yl)-3 -methoxypiperidin-4-yl)-8azabicyclo[3.2.1]octane-8-carboxylate ( 7.00 g, 16.6 mmol) fractionated 100 mg at a time by supercritical liquid chromatography using CHIRALPAK® IC/SFC (3x25 cm) using Daicel (mobile phase: CO ₂ :methanol (90:10), 120 bar, 40°C, flow rate: 100 ml/min) to obtain upon subsequent elution the title compound (3.02 g) with a retention time of 8.45 min.

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.17-1.29 (m, 3H), 1.46 (s, 9H), 1.46-1.66 (m, 6H), 1.85-1.97 (m, 3H), 2.68 (d, J=15.0 Hz, 1H), 2.75 (td, J=12.9, 3.2 Hz, 1H), 3 .30 (s, 3H), 3.35 (brs, 1H), 4.11-4.31 (m, 2H), 4.664.76 (m, 1H), 5.10 (dt, J=14.4 , 2.6 Hz, 1H), 8.13 (s, 2H).

MS (ESI) mass/charge: 443 [M+Na]+.

Analysis conditions. Supercritical Liquid Chromatography using CHIRALPAK™ IC-3 (3.0x50 mm) with Daicel (mobile phase: CO ₂ :methanol (85:15), 40°C, flow rate: 1.2 ml/min, detection: UV (254 nm)).

Analysis results. The retention time of the title compound was 1.34 min and the optical purity was >99% ee.

(7) Synthesis of (1R,3S,5S)-3-((3S,4R)-1-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8azabicyclo[3.2.1]octane.

Trifluoroacetic acid (5.00 mL, 64.9 mmol) was added to tert-butyl-(^^^)-3-(^^)-1-(5fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl) -8-azabicyclo[3.2.1]octane-8-carboxylate (1.00 g, 2.38 mmol) and the mixture was stirred for 20 minutes at room temperature. The reaction mixture was concentrated under reduced pressure. Aqueous saturated 2n. Sodium hydroxide was added to the residue and extraction was performed with ethyl acetate (3 times). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to obtain

- 8 045865 without the title compound (620 mg).

MS (ESI) mass/charge: 321[M+H]+.

Receipt example 5.

Synthesis of ^)-2-bromo-2-cyclopropylacetamide

(1) Synthesis of ^)-3-(2-cyclopropylacetyl)-4-phenyloxazolidin-2-one.

Pivaloyl chloride (302 ml, 2470 mmol) was added to tetrahydrofuran (7000 ml) at room temperature. Cyclopropylacetic acid (238 ml, 2450 mmol) was added at room temperature and the mixture was cooled to 0°C. After adding triethylamine (350 ml) dropwise over 10 minutes, triethylamine (400 ml, total amount: 5340 mmol) was added and the mixture was stirred for 76 minutes at 0°C. Then, after adding L)-4-phenyloxazolidin-2-one (350 g, 2140 mmol) to the reaction mixture at one time, lithium chloride (109 g, 2570 mmol) was further added at one time. The reaction mixture was stirred for 18 hours at room temperature, then ethyl acetate (7000 ml) and water (3500 ml) were added and the mixture was stirred for 40 minutes at room temperature. The organic layer was separated and washed with aqueous 5% sodium hydrogen carbonate solution (3500 ml) and water (1750 ml) in that order. The resulting organic layer was concentrated to 1750 ml. The azeotropic procedure of adding ethyl acetate (2100 ml) and concentrating to 1750 ml was repeated 3 times, after which ethyl acetate (1050 ml) was added and the mixture was concentrated to 1750 ml. The resulting concentrate was stirred and n-heptane (1000 ml) was added dropwise. The resulting suspension was stirred for 10 minutes and additional n-heptane (2500 ml) was added dropwise. The liquid mixture was stirred overnight at room temperature and then further stirred for 3.5 hours at 0°C. The resulting solid was filtered using a glass filter and washed with ethyl acetate/nheptane (550 ml of a 1:3 mixture). The resulting solid was dried under reduced pressure to obtain the title compound (407 g).

^1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 0.09-0.24 (m, 2H), 0.46-0.58 (m, 2H), 1.00-1.13 (m , 1H), 2.74-2.83 (m, 1H), 2.88-2.98 (m, 1H), 4.25-4.32 (m, 1H), 4.70 (t, J =8.83 Hz, 1H), 5.45 (dd, J=8.83, 3.85 Hz, 1H), 7.287.43 (m, 5H).

(2) Synthesis of ^)-2-bromo-2-cyclopropylacetamide.

Add a 1 M dichloromethane solution of dibutyl boron triflate (500 ml, 500 mmol) dropwise to a solution of N)-3-(2-cyclopropylacetyl)-4-phenyloxazolidin-2-one (100 g, 408 mmol) in dichloromethane (1000 ml) over 40 minutes while cooling on ice and the mixture was stirred for 10 minutes while cooling on ice. After adding H^diisopropylethylamine (92 mL, 530 mmol) dropwise over 25 minutes while cooling on ice, the mixture was stirred for 1 hour while cooling on ice. The reaction mixture was then cooled to an internal temperature of -72°C in a dry ice-ethanol bath. N-bromosuccinimide (80 g, 448 mmol) was then added at one time and the mixture was stirred for 1 hour and 20 minutes in a dry ice-ethanol bath. After adding 28-30% aqueous ammonia (800 ml, 6390 mmol) and tetrahydrofuran (1000 ml), the mixture was stirred for 2 hours in a water bath. The organic layer and the aqueous layer were separated, and the aqueous layer was extracted 3 times with ethyl acetate (500 ml). The resulting organic layer was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (4 kg silica gel, 30-40% ethyl acetate/n-heptane) to obtain 132 g of crude product. After adding tert-butyl methyl ether (1440 ml) to the resulting crude product and stirring for 1 hour at 50°C to dissolve, the mixture was further stirred for 1 day at room temperature. The resulting solid was filtered and washed with t-butyl methyl ether (200 ml). The filtrate and wash solution were combined, ethyl acetate (700 ml) and activated carbon (SEISEI SHIRASAGI, 26 g) were added and the mixture was stirred for 30 minutes at room temperature. The activated carbon was removed by filtration through Celite® and the activated carbon was washed with ethyl acetate (700 ml). After combining the filtrate and the washing solution, the mixture was concentrated under reduced pressure to obtain the title compound (97.1 g, 65.9% content).

1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 0.40-0.52 (m, 1H), 0.63-0.73 (m, 1H), 0.77-0.86 (m, 1H), 0.86-0.97 (m, 1H), 1.41-1.50 (m, 1H), 3.59-3.83 (m, 1H), 5.34-5.71 ( m, 1H), 5.94-6.30 (m, 1H).

MS (ESI) mass/charge: 180 [M+H]+.

Analysis conditions. Chromatography using CHIRALPAK™ IA (0.46x25 cmx2) with Daicel (mobile phase: ethanol:n-hexane (10:90), 40°C, flow rate: 0.8 ml/min, detection: UV (210 nm )).

Analysis results. The retention time of the title compound was 24.5 minutes.

Example 1.

Synthesis of (K)-2-cyclopropyl-2-((1K^^)-3-(^,4K)-1-(5-fluoropyrimidin-2-yl)-3-methoxy- 9 045865 piperidin-4-yl) -8-azabicyclo[3.2.1]octan-8-yl)acetamide

After adding 2-bromo-2-cyclopropylacetamide (433 mg, 2.43 mmol), cesium carbonate (793 mg, 2.43 mmol) and silver oxide (564 mg, 2.43 mmol) to the solution of (1R,3S,5S )-3-((3S,4R)-1-(5-fluoropyrimidin2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octane (260 mg, 0.811 mmol) in acetonitrile (4 .00 ml), the mixture was stirred for 40 hours at room temperature. The reaction mixture was purified by forward column chromatography (silica gel, 20% methanol/ethyl acetate). The resulting product was dissolved in methanol (10 ml) and fed into Waters Porapak Rxn™ CX (two 2 g cartridges). Each solid phase was washed with methanol (20 ml), then each product was eluted with ammonia (2 mol/l methanol solution, 20 ml) and the eluates were combined and concentrated under reduced pressure. The residue was dissolved in methanol and fractionated by supercritical liquid chromatography using CHIRALPAK™ (OD-H/SFC (2x25 cm) using Daicel (mobile phase: OD ₂ :methanol (85:15), 120 bar, 40°C, speed flow: 70 ml/min), at 10 mg/500 μl (methanol) per cycle to obtain a compound with a retention time of 6.41 minutes (120 mg) as an enantiomeric mixture. The resulting compound was dissolved in methanol and fractionated using supercritical liquid chromatography using CHIRALPAK™ (IG/SFC (2x25 cm) with Daicel (mobile phase: CO ₂ :methanol (75:25), 120 bar, 40°C, flow rate: 70 ml/min) at 13 mg/500 µl (methanol) per cycle to produce a compound consisting primarily of a subsequently eluting component with a retention time of 9.11 minutes (75 mg).The resulting compound was dissolved in methanol and fractionated by supercritical liquid chromatography using CHIRALPAK™ (IG/SFC ( 2x25 cm) with Daicel (mobile phase: OD ₂ :methanol (75:25), 120 bar, 40°C, flow rate: 70 ml/min) at 4 mg/100 µl (methanol) per cycle to obtain the compound, consisting mainly of a subsequently eluting component, with a retention time of 7.52 minutes (52 mg). The resulting compound was dissolved in methanol and fractionated by supercritical liquid chromatography using CHIRALPAK™ (IG/SFC (2x25 cm) using Daicel (mobile phase: CO2:methanol (75:25), 120 bar, 40°C, flow rate: 70 ml/min) at 7 mg/500 µl (methanol) per cycle to obtain the title compound, consisting mainly of the subsequently eluting component, with a retention time of 7.18 min (31.3 mg).

1H NMR (400 MHz, CDClj δ (ppm): 0.28-0.37 (m, 1H), 0.45-0.52 (m, 1H), 0.53-0.58 (m, 1H) , 0.60-0.68 (m, 1H), 0.78 (dd, J=8.8, 4.3 Hz, 1H), 1.20-1.27 (m, 2H), 1.32 -1.45 (m, 2H), 1.46-1.52 (m, 2H), 1.59-1.65 (m, 3H), 1.70-1.80 (m, 2H), 1 ,81-1.88 (m, 1H), 2.08 (d, J=9.l Hz, 1H), 2.68 (dd, J=14.3, 1.1 Hz, 1H), 2, 75 (td, J=12.8, 2.9 Hz, 1H), 3.21-3.24 (m, 1H), 3.30 (s, 3H), 3.36 (brs, 1H), 3 .86-3.91 (m, 1H), 4.65-4.76 (m, 1H), 5.10 (dt, J=14.2, 2.4 Hz, 1H), 5.14-5 .24 (m, 1H), 6.92-7.02 (m, 1H), 8.14 (s, 2H).

MS (ESI) mass/charge: 418 [M+H]+.

Analysis conditions. Chromatography using CHIRALPAK™ IA (0.46x15 cm) with Daicel (mobile phase: ethanol: hexane (20:80), 40°C, flow rate: 1 ml/min, detection: UV (254 nm)).

Analysis results. The retention time of the title compound was 4.91 min and the optical purity was >99% ee.

Preparation of single crystal (R)-2-cyclopropyl-2-((1R,3S,5S)-3-((3S,4R)-1-(5-fluoropyrimidin-2-yl)3-methoxypiperidin-4-yl)- 8-azabicyclo[3.2.1]octan-8-yl)acetamide and X-ray crystallographic analysis.

The title compound obtained in Example 1 (2.97 mg) was dissolved in methanol (1 ml). After placing 500 μl of the solution into the vial, the cap was carefully closed (a method for evaporating the solvent). After 1 day, a single crystal of the title compound was obtained in a vial. The resulting single crystal was subjected to crystallographic analysis using X-rays under the following conditions. The X-ray crystal structure of the title compound is shown in FIG. 1.

Analytical instrument: XtaLAB PRO P200 MM007HF (Rigaku, Japan).

Software: CrysAlisPro (Rigaku Oxford Diffraction).

X-ray: Monochromatic Cu-Ka with multilayer mirror (40 V/30 mA). Measurement: method of oscillation of the ω axis.

Camera length: 35 mm.

Measurement temperature: -170°C.

According to IUPAC ^)-2-cyclopropyl-2-((^^,5 S)-3 -((3 S,4R)-1 -(5-fluoropyrimidin-2-yl)-3 methoxypiperidin-4-yl)- The 8-azabicyclo[3.2.1]octan-8-yl)acetamide synthesized in Example 1 is called (2R)-2-cyclopropyl-2-{ (1R,3S,5 S)-3-[(3 S,4R )-1 -(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4yl]-8-azabicyclo[3.2.1]octan-8-yl} acetamide.

- 10 045865

Example 2.

Synthesis of (R)-2-((1R,3S,5S)-3-((3S,4R)-1-(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8azabicyclo[3.2. 1]octan-8-yl)-3-methylbutanamide

After adding ^)-2-bromo-3-methylbutanamide (326 mg, 1.81 mmol), cesium carbonate (590 mg, 1.81 mmol) and silver oxide (419 mg, 1.81 mmol) to the solution (1R, 3s,5S)-3-((3S,4R)-1-(5-fluoropyrimidin2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octane (290 mg, 0.905 mmol) in acetonitrile (8.00 ml), the mixture was stirred for 40 hours at room temperature. The reaction mixture was filtered through silica gel and the residue was washed with 20% methanol/ethyl acetate. The filtrate was concentrated under reduced pressure. The residue was dissolved in methanol (5 ml) and fed to Waters Porapak Rxn™ CX (4 g batch). The solid phase was washed with methanol (40 ml), then the product was eluted with ammonia (2N methanol solution, 40 ml) and the eluate was concentrated under reduced pressure. The residue was fractionated by supercritical liquid chromatography using CHIRALPAK™ (IG/SFC (2x25 cm) using Daicel (mobile phase: OT ₂ :methanol (80:20), 120 bar, 40°C, flow rate: 70 ml/min ) at 5 mg per cycle to give the title compound with a retention time of 4.41 min (154 mg).

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 0.95 (d, J=6.8 Hz, 3H), 1.02 (d, J=6.8 Hz, 3H), 1.15 -1.56 (m, 8H), 1.75 (td, J=10.8, 6.1 Hz, 3H), 1.86-2.10 (m, 2H), 2.67 (d, J =13.1 Hz, 1H), 2.75 (td, J=12.9, 3.2 Hz, 1H), 2.95 (d, J=4.1 Hz, 1H), 3.19-3 .27 (m, 1H), 3.30 (s, 3H), 3.33-3.46 (m, 2H), 4.70 (dt, J=13.3, 2.2 Hz, 1H), 5.06-5.16 (m, 1H), 5.30-5.36 (m, 1H), 6.79 (brd, J=5.0 Hz, 1H), 8.14 (s, 2H) .

MS (ESI) mass/charge: 421 [M+H]+.

Example 3.

Synthesis of (R)-2-(( 1 R,3S,5S)-3 -((3 S,4R)-1 -(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8azabicyclo[ 3.2.1]octan-8-yl)-2-cyclopropylacetamide

(1) Synthesis of tert-butyl-(1R,3s,5S)-3-((3S,4R)-3-methoxypiperidin-4-yl)-8-azabicyclo[3.2.1]octane-8carboxylate hemi(2S,3S )-2,3-bis-((4-methylbenzoyl)oxy)succinate.

A mixture of acetic acid (160 ml) with tert-butyl-(^^^)-3-(3-methoxypyridin-4-yl)-8azabicyclo[3.2.1]octane-8-carboxylate (20 g, 62.8 mmol) and 10% palladium on carbon (type AD, approximately 50% aqueous, 6 g, product of Kawaken Fine Chemicals Co., Ltd.) were stirred overnight at 70°C under a hydrogen atmosphere. Palladium on carbon was removed by filtration, washed with methanol (10 times the volume) and the filtrate was concentrated approximately 2 times. After adding isopropyl acetate (15 times volume) and n-heptane (3 times volume) to the resulting concentrate, the mixture was washed with 48% aqueous sodium hydroxide solution (54.7 ml). It was then concentrated approximately 5 times after adding n-heptane (100 ml) and washing with water (5 times the volume). The azeotropic procedure of adding isopropyl acetate (5 times the volume) to the resulting concentrate and concentrating approximately 5 times was repeated twice to obtain tert-butyl-(1I^^)-3-(3methoxypiperidin-4-yl)-8-azabicyclo[3.2.1 ]octane-8-carboxylate (15.1 g) as an isopropyl acetate solution.

After adding (-)-di-para-toluoyl^-tartaric acid (2.70 g, 6.98 mmol) to the solution of the resulting tert-butyl-(^^^)-3-(3-methoxypiperidin-4-yl) -8-azabicyclo[3.2.1]octane-8-carboxylate (7.55 g, 23.3 mmol) and (+)-dipivaloyl-tartaric acid (1.85 g, 5.82 mmol) in isopropyl acetate (151 ml), acetonitrile (30.2 ml) and methanol (38 ml), the mixture was stirred for 3 days at room temperature to obtain a solid.

The resulting solid was filtered and then washed with a mixed solution of isopropyl acetate and acetonitrile (9/1). The resulting stock solution was concentrated 10 times and isopropyl acetate (10 times the volume), aqueous 5N was added. sodium hydroxide solution (4.65 ml,

- 11 045865

23.269 mmol) and water (4 times the volume). After removing the aqueous layer, it was washed with water (4 times the volume). Then, after concentrating by approximately 5 times, isopropyl acetate (10 times the volume) was added and azeotroped by approximately 5 times, (+)-di-para-toluoyl D-tartaric acid (3.15 g, 8.144 mmol) was added to the solution (+) -dipivaloyl-E-tartaric acid (1.852 g, 5.817 mmol) in methanol (38 ml), acetonitrile (30.2 ml) and isopropyl acetate (151 ml) and the mixture was stirred overnight at room temperature.Then compounds (+) were added -di-para-toluoylD-tartaric acid (0.05 eq.) and isopropyl acetate (10 times the volume) The total amount was recovered by concentration, neutralization with sodium hydroxide and washing with purified water, then adding (+)-di-para-toluoyl -E-tartaric acid (3.15 g, 8.144 mmol) in a solution of (+)-dipivaloyl-Etartaric acid (0.2 eq.) in isopropyl acetate (151 ml), acetonitrile (30.2 ml) and methanol (38 ml ) and the mixture was stirred overnight at room temperature.(+)-Di-para-toluoylD-tartaric acid compound (0.05 eq.) was added to the reaction mixture and the mixture was stirred for 4 hours at room temperature. Compounds of (+)-di-para-toluoyl-E-tartaric acid (0.025 eq.) and isopropyl acetate (10 times volume) were added to the reaction mixture and the mixture was stirred overnight at room temperature. The total amount was recovered by concentration, neutralization with sodium hydroxide and washing with purified water, adding (+)-di-para-toluoyl-E-tartaric acid (3.15 g, 8.14 mmol) to the (+)-dipivaloyl-E- solution tartaric acid (0.2 eq.) in isopropylacetate (151 ml), acetonitrile (30.2 ml) and methanol (38 ml) and the mixture was stirred overnight at room temperature. Isopropyl acetate was then added (10 times the volume). After 7 hours, the resulting solid was filtered and washed with isopropyl acetate to give the title compound (3.45 g).

Analysis conditions. Chromatography using CHIRALPAK™ IA (0.46x25 cm) with Daicel (mobile phase: ethanol:n-hexane (20:80), 40°C, flow rate: 1.0 ml/min, detection: UV (254 nm )).

Analysis results. The retention time of the title compound was 8.52 min and the optical purity was 99.2% ee.

(2) Synthesis of tert-butyl-(1R.3s.5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-methoxypiperidin-4-yl)8-azabicyclo [3.2.1]octane-8-carboxylate.

Ν,N-Dimethylformamide (10 ml) containing 2,5-dichloropyrimidine (288 mg, 1.93 mmol), tert-butyl-(^^^)-3-(^^)-3-methoxypiperidin-4-yl )-8-azabicyclo[3.2.1]octane-8-carboxylate hemi(2S,3S)-2,3-bis-((4-methylbenzoyl)oxy)succinate (500 mg, 0.966 mmol) and potassium carbonate (267 mg , 1.93 mmol) was stirred for 24 hours at 80°C. The reaction mixture was cooled to room temperature, then water was added and extraction was performed 3 times with ethyl acetate. The organic layer was collected and concentrated. The resulting residue was purified by column chromatography (silica gel, 10-100% ethyl acetate/n-heptane) to give the title compound (368 mg).

1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.24-1.27 (m, 2H), 1.41-1.49 (m, 2H), 1.46 (s, 9H), 1 .61 (brs, 4H), 1.82-1.98 (m, 4H), 2.68 (d, J=14.04 Hz, 1H), 2.75 (td, J=12.91, 3 .17 Hz, 1H), 3.30 (s, 3H), 3.36 (brs, 1H), 4.114.32 (m, 2H), 4.68-4.80 (m, 1H), 5.12 (dt, J=14.27, 2.61 Hz, 1H), 8.16 (s, 2H).

(3) Synthesis of tert-butyl-(1R,3s,5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-hydroxypiperidin-4-yl)8-azabicyclo[3.2 .1]octane-8-carboxylate.

Mixture of Ίret-butyl-(1R,3s,5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8azabicyclo[3.2.1]octane -8-carboxylate (300 mg, 0.687 mmol) and 48% hydrobromic acid (5 ml) were stirred for 20 minutes at room temperature and then heated for 2.5 hours at 100°C. It was then stirred overnight at room temperature and the resulting reaction mixture was concentrated under reduced pressure. To the resulting residue were added tetrahydrofuran (10 ml) and an aqueous saturated sodium hydrogen carbonate solution, and then di-tert-butyl dicarbonate (165 mg, 0.755 mmol) was added at room temperature. After confirming the completion of the reaction by UPLC, ethyl acetate and water were added and the organic layer was separated. The aqueous layer was again extracted with ethyl acetate, combined with the previously obtained organic layer, and dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The resulting residue was washed with a mixed solvent of 10% ethyl acetate/n-heptane to obtain the title compound (226 mg).

1H NMR (400 MHz, CDC13) δ (ppm): 1.28-1.34 (m, 2H), 1.46 (s, 9H), 1.59-1.72 (m, 6H), 1. 75-1.97 (m, 4H), 2.70-2.83 (m, 1H), 2.91 (dd, J=14.04, 0.91 Hz, 1H), 3.97-4, 03 (m, 1H), 4.09-4.34 (m, 2H), 4.68-4.79 (m, 1H), 4.81-4.91 (m, 1H), 8.18 ( s, 2H).

MS (ESI) mass/charge: 423 [M+H]+.

(4) Synthesis of tert-butyl-(1RAsAS)^-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8azabicyclo[3.2.1]octane -8-carboxylate.

Sodium hydride (60%, dispersed in liquid paraffin, 7.09 mg, 0.177 mmol) was added to a solution of tert-butyl-(1R,3s,5S)-3-((3S,4R)-1-(5-chloropyrimidine- 2-yl)-3-hydroxypiperidin-4-yl)-8azabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.118 mmol) in N,N-dimethylformamide (1 mL) with ice cooling. After stirring for 30 min, ethyl iodide (0.019 ml, 0.236 mmol) was added and

- 12 045865 the mixture was stirred for 16 hours at room temperature. After water and ethyl acetate were added to the reaction mixture, the organic layer was separated and concentrated. The resulting residue was purified by column chromatography (silica gel, 1-20% ethyl acetate/n-heptane) to give the title compound (42.5 mg).

MS (ESI) mass/charge: 451 [M+H]+.

(5) Synthesis of (R)-2-((1R,3S,5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8azabicyclo [3.2.1]octan-8-yl)-2-cyclopropylacetamide.

After treatment tert-butyl-(1R,3s,5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4yl)-8-azabicyclo[3.2.1] octane-8-carboxylate (42.5 mg, 0.094 mmol) with trifluoroacetic acid (1 ml) over 30 min, the mixture was concentrated. The resulting residue was dissolved in methanol and added to Waters Porapak Rxn™ CX (0.4 g). The solid phase was washed with methanol (6 ml), then the product was eluted with ammonia (2 mol/l methanol solution, 6 ml) and the eluate was concentrated under reduced pressure. A mixture of the resulting residue ^)-2-bromo-2-cyclopropylacetamide (51.6 mg, 0.188 mmol) obtained by the same procedure as Preparation Example 5, potassium carbonate (26.0 mg, 0.188 mmol) and acetonitrile (2 ml) was stirred for 17 days at room temperature. The reaction mixture was filtered using acetonitrile (3 ml) and the resulting filtrate was fractionated using supercritical liquid chromatography using CHIRALPAK® (IA/SFC (3x25 cm) using Daicel (mobile phase: CO ₂ :methanol (75:25), 120 bar , 40°C, flow rate: 100 ml/min) at 500 µl per cycle to obtain the title compound with a retention time of 7.55 min (22.2 mg).

1H NMR (400 MHz, CDCl3) δ (ppm): 0.26-0.38 (m, 1H), 0.40-0.58 (m, 2H), 0.59-0.68 (m, 1H ), 0.71-0.83 (m, 1H), 0.98-1.92 (m, 12H), 1.04 (t, J=7.02 Hz, 3H), 2.07 (brd, J=9.06 Hz, 1H), 2.61-2.81 (m, 2H), 3.13-3.30 (m, 2H), 3.45 (brs, 1H), 3.61-3 .75 (m, 1H), 3.84-3.92 (m, 1H), 4.66-4.76 (m, 1H), 4.93-5.11 (m, 1H), 5.17 -5.32 (m, 1H), 6.90-7.03 (m, 1H), 8.16 (s, 2H).

MS (ESI) mass/charge: 448 [M+H] ⁺ .

Analysis conditions. Supercritical Liquid Chromatography using CHIRALPAK™ IA-3 (0.46x25 cm) with Daicel (mobile phase: CO ₂ :methanol (80:20), 40°C, flow rate: 1.2 ml/min, detection: UV (257 nm)).

Analysis results. The retention time of the title compound was 2.19 min and the optical purity was >99.9% e. And.

Preparation of single crystal (R)-2-((1R,3S,5S)-3-((3S,4R)-1-(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8-azabicyclo [3.2.1]octan-8-yl)-2-cyclopropylacetamide and X-ray crystallographic analysis.

The title compound obtained in Example 3 (1.37 mg) was dissolved in acetonitrile (600 μl). After placing 200 μl of the solution into the vial, the cap was carefully closed (a method for evaporating the solvent). After 1 day, a single crystal of the title compound (a dimer crystal with two added acetonitrile groups) was obtained in a vial. The resulting single crystal was subjected to crystallographic analysis using X-rays under the following conditions. The X-ray crystal structure of the title compound is shown in FIG. 2.

Analytical instrument: XtaLAB PRO P200 MM007HF (Rigaku, Japan).

Software: CrysAlisPro (Rigaku Oxford Diffraction).

X-ray: Monochromatic Cu-Ka with multilayer mirror (40 V/30 mA).

Measurement: method of oscillation of the ω axis.

Camera length: 35 mm.

Measurement temperature: -170°C.

Example 4.

Synthesis of (R)-2-cyclopropyl-2-((1R,3S,5S)-3-((2S,4S)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin4-yl)-8- azabicyclo[3.2.1]octan-8-yl)acetamide

O

(1) Synthesis of ethyl-(^^^)-3-(^)-2-methyl-1,2,3,6-tetrahydropyridin-4-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate .

Added to a 13 mm diameter test tube with a screw cap

- 13 045865 ethyl-(1P+^^)-3-(tosyloxy)-8-azabicyclo[3.2.1|octane-8-carboxylate (CAS No. 2236076-85-8) (200 mg, 0.566 mmol), complex Nickel(II) bromide and ethylene glycol dimethyl ether (17.5 mg, 0.057 mmol), 4,4-di-1ret-butyl-2,2-dipyrrvdyl (15.2 mg, 0.057 mmol), potassium iodide (94.0 mg, 0.566 mmol) and manganese (62.2 mg, 1.13 mmol). Then a solution of tert-butyl-^)-2-methyl-4-(((trifluoromethyl)sulfonyl)oxy)-3,6-dihydropyridine-1(2H)-carboxylate (CAS No. 876922-74-6) was added (195 mg, 0.566 mmol) in Ν,N-dimethylacetamide (4.0 ml) and 4-ethylpyridine (0.064 ml, 0.566 mmol) under a stream of nitrogen. The resulting mixture was stirred for 12.5 hours at 80°C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and then diluted with ethyl acetate. Insoluble materials were removed by filtration through a cotton plug and the eluate was partitioned between water and ethyl acetate. The aqueous layer was separated and extracted with ethyl acetate. The organic layers were combined, washed 3 times with water, concentrated under reduced pressure, and the coupling reaction product was obtained as a crude product.

Trifluoroacetic acid (1.0 ml) was added to a solution of the resulting crude product in dichloromethane (4.0 ml) and the mixture was stirred for 1 hour at room temperature. The reaction mixture was purged with nitrogen to distill off the solvent, and the residue was diluted with methanol. The resulting methanol solution was fed into the Waters PoraPak Rxn™ CX (20 cc (2 g) cartridge). After washing the solid phase with methanol (20.0 ml), it was eluted with ammonia (2N methanol solution, 20 ml). The eluate was concentrated under reduced pressure to give the crude product of the title compound (120 mg).

MS (ESI) mass/charge: 279 [M+H]+.

(2) Synthesis of ethyl-(^^^)-3-(^^)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octane- 8-carboxylate.

Mixture of ethyl-(^^^)-3-(^)-2-methyl-1,2,3,6-tetrahydropyridin-4-yl)-8-azabicyclo[3.2.1]octane-8carboxylate (120 mg, 0.431 mmol), 10% palladium on carbon (48.47% aqueous product solution, 183 mg, 0.083 mmol, product of N.E. Chemcat Corp.) and methanol (4.0 ml) were stirred for 6.5 hours under a hydrogen atmosphere. Insolubles were removed by filtration through Celite™ and concentrated under reduced pressure to give the reduced form (113 mg) as a cis/trans mixture.

MS (ESI) mass/charge: 281 [M+H]+.

A mixture of the resulting reduced form (113 mg), 2-chloro-5-fluoropyrimidine (0.048 ml, 0.517 mmol), cesium carbonate (281 mg, 0.862 mmol) and N,N-dimethylacetamide (2.0 ml) was stirred for 8 h at 100°C. The mixture was purified by column chromatography (silica gel, 0-30% ethyl acetate/n-heptane) to give the title compound (82.0 mg) as a cis/trans mixture.

MS (ESI) mass/charge: 377 [M+H]+.

Analysis conditions. Supercritical Liquid Chromatography using CHIRALPAK™ IF-3 (3.0x50mm) with Daicel (mobile phase: CO ₂ :methanol (70:30), 40°C, flow rate: 1.2 ml/min, detection: UV (210-400 nm)).

Analysis results. The retention time of the title compound was 1.02 minutes, and the retention time of the trans form was 1.23 minutes. The cis:trans ratio was 4:5 (peak area ratio) and the optical purity was >99% ee.

The resulting cis/trans mixture was fractionated by supercritical liquid chromatography using CHIRALPAK® (IF/SFC (3x25 cm) using Daicel (mobile phase: CO ₂ :methanol (70:30), 120 bar, 40°C, flow rate: 100 ml/min) at 10 mg per cycle to obtain the first eluting title compound with a retention time of 5.55 min (34.0 mg).

Ή NMR (400 MHz, CDCl ₃ ) δ (ppm): 1.17 (d, J=6.34 Hz, 3H), 1.23 (t, J=7.25 Hz, 3H), 1.19- 2.02 (m, 14H), 3.09 (ddd, J=13.70, 10.99, 5.66 Hz, 1H), 4.10 (q, J=7.10 Hz, 2H), 4 .17-4.30 (m, 3H), 4.34 (dd, J=13.59, 7.25 Hz, 1H), 8.13 (s, 2H).

MS (ESI) mass/charge: 377 [M+H]+.

Analysis conditions. Supercritical Liquid Chromatography using CHIRALPAK™ IF-3 (3.0x50mm) with Daicel (mobile phase: CO ₂ :methanol (70:30), 40°C, flow rate: 1.2 ml/min, detection: UV (245 nm)).

Analysis results. The retention time of the title compound was 1.02 min, the cis:trans ratio was >99:1, and the optical purity was >99% ee.

(3) Synthesis of (^^^)-3-(^^)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octane.

A mixture of ethyl-(1R,3s,5S)-3-((2S,4S)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octane -8-carboxylate (34 mg, 0.09 mmol) and 48% hydrobromic acid (2.0 ml) were stirred for 1 hour at 100°C. For neutralization, aqueous 5N was added to the reaction mixture at 0°C. sodium hydroxide solution. The mixture was extracted 3 times with dichloromethane, and the organic layer was dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain the title compound (21.7 mg).

MS (ESI) mass/charge: 305 [M+H]+.

- 14 045865 (4) Synthesis of ^)-2-cyclopropyl-2-((^^^)-3-(^^)-1-(5-fluoropyrimidin-2-yl)-2-methyl-pipervdin-4- yl)-8-azabicyclo[3.2.1]octan-8-yl)acetam.

A mixture of ( ^1I.p ^^)-3-((^^)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8-azabicyclo[3.2.1]octane (5, 00 mg, 0.016 mmol), potassium carbonate (4.54 mg, 0.033 mmol), (S)-2-6poM-2cyclopropylacetamide (8.24 mg, 0.033 mmol) and acetonitrile (1.0 ml) were stirred at room temperature in within 7 days. Potassium carbonate (4.54 mg, 0.033 mmol) and N-2-bromo-2-cyclopropylacetamide (8.24 mg, 0.033 mmol) were added to the reaction mixture, which was then stirred at room temperature for 3 days. An aqueous ammonium chloride solution was added to the reaction mixture to suspend the reaction mixture. The mixture was extracted with ethyl acetate, the organic layer was washed with water and concentrated under reduced pressure to obtain the crude product.

The resulting crude product was diluted with methanol and fed into a Waters PoraPak Rxn™ CX (6 cc (400 mg) cartridge). After washing the solid with methanol (6.0 ml), it was eluted with ammonia (2N methanol solution, 6 ml). The eluate was concentrated under reduced pressure and the resulting residue was purified by thin layer chromatography (NH, dichloromethane) to give the title compound (3.74 mg).

^1H NMR (400 MHz, CDCI3) δ (ppm): 0.25-0.36 (m, 1H), 0.39-0.51 (m, 1H), 0.51-0.59 (m, 1H), 0.59-0.70 (m, 1H), 0.70-0.84 (m, 1H), 1.18 (d, J=6.34 Hz, 3H), 1.21-1 .55 (m, 9H), 1.61-1.95 (m, 5H), 2.04 (d, J=9.06 Hz, 1H), 3.10 (ddd, J=13.70.10 .99, 5.21 Hz, 1H), 3.15-3.28 (m, 1H), 3.80-3.94 (m, 1H), 4.17-4.29 (m, 1H), 4.35 (dd, J=13.59, 7.25 Hz, 1H), 5.18 (brs, 1H), 6.94 (brs, 1H), 8.14 (s, 2H).

MS (ESI) mass/charge: 402 [M+H]+.

Preparation of a single crystal of ^)-2-cyclopropyl-2-((^^^)-3-(^^)-1-(5-fluoropyrimidin-2-yl)2-methylpiperidin-4-yl)-8-azabicyclo[3.2 .1]octan-8-yl)acetamide and X-ray crystallographic analysis.

The title compound obtained in Example 4 (0.81 mg) was dissolved in methanol (600 μL). The vial containing the 200 µl portion of it was carefully placed into a slightly larger vial containing 2 ml of tert-butyl methyl ether and capped (vapor diffusion method). After 12 days, a single crystal of the title compound was obtained in a vial. The resulting single crystal was subjected to crystallographic analysis using X-rays under the following conditions. The X-ray crystal structure of the title compound is shown in FIG. 3.

Analytical instrument: XtaLAB PRO P200 MM007HF (Rigaku, Japan).

Software: CrysAlisPro (Rigaku Oxford Diffraction).

X-ray: Monochromatic Cu-Κα with multilayer mirror (40 V/30 mA). Measurement: method of oscillation of the ω axis.

Camera length: 35 mm.

Measurement temperature: -170°C.

Examples of pharmacological tests.

The following pharmacological test was carried out using the compounds of Examples 1-4. Test case 1-1. Assessment of activity against OX1R and OX2R.

HEK293 (human embryonic kidney 293) cells forced to express human OX1R (hOX1R) or human OX2R (hOX2R) were seeded in 384-well microplates (Greiner) at 10,000 per well and cultured for 1 day in high glucose DMEM (FujiFilm Corp.-Wako Pure Chemical Industries Ltd.) containing added 10% FBS (Thermo Scientific) and 1% penicillin-streptomycin (FujiFilm Corp.-Wako Pure Chemical Industries, Ltd.). After removing the medium, 40 μl of assay buffer (20 mM HEPES (Sigma-Aldrich Japan, KK.), Hanks' balanced salt solution (Gibco), 0.1% BSA (Sigma-Aldrich Japan, KK.), 0.1% Pluronic F-127 (Invitrogen) containing Calcium 4 dye (Molecular Device Corporation) and 2.5 mM probenecid (Sigma-Aldrich Japan, KK.), and the plate was incubated for 60 min. After an additional 20 μl of assay buffer was added, 20 μl of assay buffer containing the test compound was added and the reaction was initiated. The change in intracellular calcium ion concentration during the reaction was measured based on the fluorescence intensity ratio according to the fluorescence value with excitation at 480 nm and detection at 540 nm using an FDSS7000 (Hamamatsu Photonics, KK). The test compound was dissolved in DMSO to a concentration of 10 mM and diluted with assay buffer to a final concentration of 3x10 ^-10 to 1x10 ^-7 M (final DMSO concentration 0.1%). The half-maximal effective concentration (EC ₅₀ value) was determined by the fluorescence value when adding the test compound at various concentrations, while the fluorescence value in the well with the added buffer containing no compound was taken as 0%, and the fluorescence value in the well with OX-A concentration , 10 nM (Peptide Research Lab), was taken as 100%. The EC ₅₀ value of each compound is shown in Table. 1.

- 15 045865

Table 1

Example No. hOXlREC ₅₀ (nM) hOX2R EC ₅₀ (nM) 1 >100 0.99 2 >100 3.08

Test case 1-2. Evaluation of activating activity against OX1R and OX2R.

Human embryonic kidney 293 (HEK293) cells forced to express hOX1R or hOX2R were seeded in 384-well microplates (Greiner) at 10,000 per well and cultured for 1 day in high glucose DMEM (FujiFilm Corp.-Wako Pure Chemical Industries Ltd .) containing added 10% FBS (Thermo Scientific) and 1% penicillin-streptomycin (FujiFilm Corp.-Wako Pure Chemical Industries, Ltd.). After removing the medium, 30 μl of assay buffer (20 mM HEPES (Sigma-Aldrich Japan, KK.), Hanks' balanced salt solution (Gibco), 0.1% BSA (Sigma-Aldrich Japan, KK.), 0.1% Pluronic F-127 (Invitrogen) containing Calcium 4 dye (Molecular Device Corporation) and 2.5 mM probenecid (Sigma-Aldrich Japan, KK.), and the mixture was incubated for 60 min. A 30-μl portion of the assay buffer containing the test compound was added and the reaction was initiated. The change in intracellular calcium ion concentration due to the reaction was measured based on the fluorescence intensity ratio according to the fluorescence value with excitation at two wavelengths of 480 and 540 nm using an FDSS7000 (Hamamatsu Photonics, KK). The test compound was dissolved in DMSO to a concentration of 10 mM and diluted with assay buffer to a final concentration of 3 x 10 ^-11 to 1 x 10 ^-5 M (final DMSO concentration of 0.1%). The EC ₅₀ value was determined by the fluorescence value when adding the test compound at various concentrations, while the fluorescence value in the well with the added buffer containing no compound was taken as 0%, and the fluorescence value in the well with OX-A at a concentration of 30 nM ( Peptide Research Lab), was taken as 100%. The EC ₅₀ value of each compound is shown in Table. 2.

table 2

Example No. hOXlREC ₅₀ (nM) hOX2R EC ₅₀ (nM) 1 4700 2.3 3 1400 4.3 4 3900 4.4

Test case 2. Increased spontaneous movement.

Similar to an increase in body temperature or an increase in cardiovascular parameters such as blood pressure, increased movement in mice is one of the indicators of wakefulness. In this test example, the alertness effect was assessed by measuring the spontaneous movement of the mice. Male C57BL/6NCrl mice were used for the experiment (age 18-19 weeks, Charles River Laboratories, Japan Inc., 4 mice in each group). Spontaneous movement was measured using a movement measuring device (VersaMax Oven Field, AccuScan Instruments, Inc.) by irradiating the side sections of the measuring cage with infrared rays and quantifying the number of times the mice passed through the irradiation. After placing the mice in the measuring cage and conditioning for 3 hours, the compound (10 mg/kg) was administered orally. Spontaneous movement was measured 2 h after administration. The test compound-administered group was administered a solution of the test compound dissolved in a 0.1 mol/L hydrochloric acid solution containing 5% (v/v) DMSO and 5% (v/v) Kolliphor™ EL. In the control group, mice were administered only the vehicle without the test compound.

The results are shown in table. 3. Spontaneous movement was reported as a percentage for the group administered the test compound, and spontaneous movement in the control group was taken as 100%.

Table 3

Compound under test Spontaneous movement (% control) solvent 100 Example 1 865 Example 2 253

As shown in table. 3, the compounds of the present invention increased spontaneous movement in mice. In other words, the compounds of the present invention have been shown to have an alerting effect.

Test Example 3: Wake-promoting effect of oral administration of the compound of the present invention to wild-type mice.

Male wild-type (WT) C57BL/6 mice were used as experimental animals. Electrodes for measuring electroencephalogram and electromyogram were inserted surgically

- 16 045865 in 13-week-old mice under isoflurane anesthesia. The solvent (0 mg/kg) or a solution of the compound from Example 1 dissolved in the solvent (1.3 or 10 mg/kg) was administered orally 30-15 minutes before turning off the lights. The solvent used was 0.1 mol/L hydrochloric acid solution containing 5% (v/v) DMSO and 5% (v/v) Kolliphor™ EL. The electroencephalogram and electromyogram were recorded for approximately 24 hours, starting 1 hour before the lights were turned off. Mice were reused with a washout period of 2 days or more. The electroencephalogram and electromyogram data obtained for each mouse were used to estimate the sleep stage of each interval (10 s) using sleep analysis software (SleepSign: Kissei Comtec Co., Ltd.). The time (sleep onset latency) to the onset of initial light-off sleep (sleeping for 8 or more intervals beginning after non-REM sleep) was measured for each mouse. Using 16 mice in each administration group, the sleep onset latency of the vehicle administration group (control group) and the compound administration group of Example 1 was compared using Dunnett's multiple comparison test after survival time was analyzed taking into account the number of experiments and administration the same individuals with a significance level of 5% on both sides for each.

Sleep latency rates in mice administered the vehicle and compound of Example 1 at doses of 1, 3, and 10 mg/kg were 0.23, 0.28, 0.44, and 2.07 hours, respectively. In the groups that were administered the compound of Example 1 at a dose of 3 or 10 mg/kg, sleep onset latency increased significantly compared to the vehicle group. In particular, it was found that when the compound of Example 1 was administered orally, sleep onset latency was prolonged in a dose-dependent manner.

Test Example 4: Awakening effect and prolongation of cataplexy-free state upon oral administration of the compound of the present invention to orexin-deficient mice (orexin/ataxin-3 Tg/+ mice).

Orexin/ataxin-3 Tg/+ mice (hereinafter referred to as Tg mice, Hara et al., Neuron, 30, 345-54, 2001) with a C57BL/6 genetic background were used as experimental animals. Electrodes for electroencephalogram and electromyogram measurements were surgically implanted into 12-week-old mice (±2 weeks) under isoflurane anesthesia. Solvent (0 mg/kg) or sample solution (compound from Example 1 dissolved in solvent: 0.3, 1 or 3 mg/kg) was administered orally 30-15 minutes before turning off the lights. The solvent used was 0.1 mol/L hydrochloric acid solution containing 5% (v/v) DMSO and 5% (v/v) Kolliphor™ EL. The electroencephalogram and electromyogram were recorded for approximately 24 hours, starting 1 hour before the lights were turned off. Mice were reused with a washout period of 2 days or more. The electroencephalogram and electromyogram data obtained for each mouse were used to estimate the sleep stage of each interval (10 s) using sleep analysis software (SleepSign: Kissei Comtec Co., Ltd.), up to a maximum of 4 h (threshold). For the purposes of this experiment, cataplexy-like symptoms were defined as REM sleep occurring immediately after wakefulness (direct transition from wakefulness to REM sleep (DREM)) for a continuous period of 4 intervals or longer. DREM in mice is analogous to cataplexy (Exp Neurol. 2009;217:46-54). For each mouse, the time to onset of initial light-off sleep (sleeping for 8 continuous intervals or longer, excluding DREM) (sleep onset latency) and the time to onset of initial DREM (DREM latency) were measured. The number of mice was 14 in the vehicle-treated group and the compound-administered groups of Example 1. The sleep onset latency and DREM latency in the vehicle-treated control group and the compound-administered group were compared using a multiplex test. Dunnett's comparison after analysis of survival time taking into account the number of experiments and the use of the same individuals with a significance level of 5% on both sides for each.

Sleep onset latencies in Tg mice treated with the vehicle and compound of Example 1 at 0.3, 1, and 3 mg/kg were 0.21, 0.31, 0.64, and 2.42 hours, respectively, indicating to a significant increase in sleep onset latency in groups that were administered the compound from Example 1 at a dose of 1 and 3 mg/kg. In particular, it was found that when the compound of Example 1 was administered orally, sleep onset latency was prolonged in a dose-dependent manner from 1 mg/kg in orexin-deficient mice.

The DREM latency values for vehicle administration and 0.3, 1, and 3 mg/kg of Example 1 in Tg mice were 1.16, 1.50, 2.26, and 4.00 hours, respectively, confirming that the latency DREM increased significantly and in a dose-dependent manner in the groups receiving the compound of Example 1 at 0.3, 1 and 3 mg/kg compared to the vehicle group. In other words, conditions without cataplexy-like behavior were maintained in mice by administering the compounds of the present invention in a dose-dependent manner.

Claims (15)

1. A connection selected from the group consisting of: (2C)-2-cyclopropyl-2-{(1 R,3 S,5S)-3 - [(3 S,4R)-1 -(5 -fluoropyrimidin-2-yl)-3 -methoxypiperidin-4yl]- 8-azabicyclo[3.2.1]octan-8-yl}acetamide represented by the following formula (I): (R)-2-(( 1R,3S,5S)-3 -((3S,4R)-1 -(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4-yl)-8-azabicyclo- [3.2.1]ok tan-8-yl)-3-methylbutanamide represented by the following formula (II): (R)-2-(( 1R,3 S,5S)-3 -((3 S,4R)-1 -(5-chloropyrimidin-2-yl)-3-ethoxypiperidin-4-yl)-8azabicyclo[3.2 .1]octan-8-yl)-2-cyclopropylacetamide represented by the following formula (III): (R)-2-cyclopropyl-2-((1R,3S,5S)-3-((2S,4S)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl)-8azabicyclo [3.2.1]octan-8-yl)acetamide represented by the following formula (IV): or a pharmaceutically acceptable salt thereof. 2. (2R)-2-Cyclopropyl-2-{(1R,3S,5S)-3-[(3S,4R)-1 -(5-fluoropyrimidin-2-yl)-3-methoxypiperidin-4yl]- 8-azabicyclo[3.2.1]octan-8-yl}acetamide represented by the following formula (I): Δ or a pharmaceutically acceptable salt thereof. 3. (R)-2-(( 1 R,3 S,5 S)-3 -((3 S,4R)-1 -(5 -Fluoropyrimidin-2-yl)-3 -methoxypiperidin-4-yl) -8-azabicyclo[3.2.1]octan-8-yl)-3-methylbutanamide represented by the following formula (II): or a pharmaceutically acceptable salt thereof. 4. (R)-2-(( 1R,3S,5S)-3 -((3 S,4R)-1 -(5-Chloropyrimidin-2-yl)-3 -ethoxypiperidin-4-yl)-8- azabicyclo- [3.2.1]oc tan-8-yl)-2-cyclopropylacetamide represented by the following formula (III): or a pharmaceutically acceptable salt thereof. 5. (R)-2-Cyclopropyl-2-((1R,3S,5S)-3-((2S,4S)-1-(5-fluoropyrimidin-2-yl)-2-methylpiperidin-4-yl) 8-azabicyclo[3.2.1]octan-8-yl)acetamide represented by the following formula (IV): - 18 045865 or a pharmaceutically acceptable salt thereof. 6. A pharmaceutical composition containing a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 7. An orexin receptor type 2 agonist, comprising a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 8. A therapeutic agent for the treatment of narcolepsy containing a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 9. A method of treating narcolepsy, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 10. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the treatment of narcolepsy. 11. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 for the manufacture of a pharmaceutical composition intended for the treatment of narcolepsy. 12. A therapeutic agent for the treatment of cataplexy, containing a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 13. A method of treating cataplexy, comprising administering to a subject a pharmacologically effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5. 14. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the treatment of cataplexy. 15. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 for the manufacture of a pharmaceutical composition intended for the treatment of cataplexy.