KR102611847B1 - Novel pyrrolidinium compounds having antagonistic activity against muscarinic receptors and use thereof - Google Patents

Abstract

The present invention relates to novel pyrrolidinium compounds, stereoisomers, hydrates or solvates thereof, which exhibit excellent binding ability and antagonistic activity to muscarinic receptors, especially muscarinic M3 receptors. The compounds of the present invention are directed to muscarinic acetylcholine receptors. It is useful in the treatment of vector diseases such as hyperhidrosis, hypersalivation, chronic obstructive pulmonary disease, chronic bronchitis, asthma, rhinitis, urinary incontinence, overactive bladder, reflux esophagitis, and irritable bowel syndrome.

Description

NOVEL PYRROLIDINUM COMPOUNDS HAVING ANTAGONISTIC ACTIVITY AGAINST MUSCARINIC RECEPTORS AND USE THEREOF}

The present invention relates to a novel pyrrolidinium compound having muscarinic receptor antagonistic activity, its stereoisomer, hydrate or solvate, a pharmaceutical composition containing it as an active ingredient, and its medicinal use.

Muscarinic acetylcholine receptors (mAChRs) belong to the superfamily of G-protein-coupled receptors, with seven transmembrane domains. There are five subtypes of mAChR, referred to as M1 to M5, each of which exhibits unique pharmacological properties. For example, in the airways, stimulation of M3 receptors causes contraction of airway smooth muscles, resulting in bronchoconstriction, and in salivary and sweat glands, stimulation of M3 receptors increases fluid and mucous membrane secretion, resulting in increased secretion of saliva and sweat.

Muscarinic receptor antagonists act by inhibiting the binding of acetylcholine to muscarinic cholinergic receptors at various points of action in the body, such as smooth muscle, myocardium, peripheral ganglia, and central nervous system. Therefore, muscarinic receptor antagonists are effective in treating various diseases, such as chronic obstructive pulmonary disease (COPD), chronic bronchitis, asthma, rhinitis, hypersalivation, hyperhidrosis, urinary incontinence, and overactive bladder. It is known to be useful in the treatment of bladder syndrome, gastroesophogeal reflux disease, and irritable bowel syndrome.

Accordingly, the present inventors completed the present invention by preparing a novel pyrrolidinium compound that exhibits excellent antagonistic activity against muscarinic receptors, especially muscarinic M3 receptors.

The object of the present invention is to provide a compound represented by formula (I), a stereoisomer, hydrate or solvate thereof.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating muscarinic acetylcholine receptor-mediated diseases, comprising the compound represented by formula (I), its stereoisomer, hydrate or solvate, and a pharmaceutically acceptable carrier. It is done.

Another object of the present invention is to provide a method for preventing or treating muscarinic acetylcholine receptor-mediated diseases, comprising the step of administering or topically applying a compound represented by Formula I, a stereoisomer, hydrate or solvate thereof to an individual. will be.

Each description and embodiment disclosed in this application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Additionally, the scope of the present application cannot be considered limited by the specific description described below.

The present invention provides compounds of formula (I), stereoisomers, hydrates or solvates thereof:

R ¹ and R ² are each independently hydrogen, halogen, hydroxy, cyano, amino, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl _, C ₁ - selected from the group consisting of C ₆ alkoxy, C ₁ -C ₆ haloalkoxy, C ₁ -C ₆ alkylamino, dialkylamino and aminocarbonyl;

R ³ is hydrogen or C ₁ -C ₆ alkyl;

R ⁴ is absent, A, -OC(O)-A, -OA or -C(O)-A;

A is 5- to 6-membered heteroaryl or 5- to 6-membered heterocycloalkyl containing 1 to 3 heteroatoms selected from N, O or S, wherein the 5- to 6-membered heteroaryl and 5-membered to 6-membered heterocycloalkyl may be optionally substituted with oxo, halogen, hydroxy, cyano, amino, C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

n and m are each independently integers from 0 to 3;

k is an integer from 1 to 3,

p is an integer from 1 to 5,

X ^- is a monovalent anion.

In Formula I of the present invention, R ¹ and R ² are each independently hydrogen, halogen, hydroxy, cyano, amino, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ It may be selected from the group consisting of hydroxyalkyl _, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy, C ₁ -C ₆ alkylamino, dialkylamino and aminocarbonyl. In some embodiments, R ¹ and R ² can each independently be hydrogen, halogen, hydroxy, cyano, or C ₁ -C ₃ alkoxy. In one embodiment, R ¹ and R ² may each independently be hydrogen or halogen. In this case, the halogen may be F, Cl, Br or I. For example, the halogen may be F or Cl. In one embodiment, R ¹ may be a halogen (eg, F, Cl, Br, I) and R ² may be hydrogen. In one embodiment, R ¹ may be hydrogen and R ² may be halogen (eg, F, Cl, Br, I). In one embodiment, R ¹ and R ² can both be hydrogen.

In Formula I, n and m are each independently 0, 1, 2, or 3. In some embodiments, n and m can each independently be 0, 1, or 2. When n and m are each independently 2 or 3, a plurality of R ¹ and R ² may be the same or different from each other.

In formula I of the present invention, R ³ is hydrogen or C ₁ -C ₆ alkyl. In one embodiment, R ³ may be C ₁ -C ₆ alkyl. For example, R ³ may be C ₁ -C ₃ alkyl. For example, R ³ may be methyl, ethyl, propyl or isopropyl.

In Formula I of the present invention, p is an integer from 1 to 5. In some embodiments, p can be 1, 2, or 3.

In formula I of the present invention, R ⁴ is absent, -A, -OC(O)-A, -OA or -C(O)-A. In this case, A is 5-6 membered heteroaryl or 5-6 membered heterocycloalkyl. The 5- to 6-membered heteroaryl or 5- to 6-membered heterocycloalkyl may include 1 to 3 heteroatoms selected from N, O, or S. For example, the 5- to 6-membered heteroaryl may include pyridine, pyrimidine, pyrrolyl, pyrazolyl, furanyl, thiophenyl, etc., but is not limited thereto. For example, the 5- to 6-membered heterocycloalkyl is tetrahydropyranyl, dihydropyranyl, 1,3-dioxolyl, 1,3-dioxolanyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, etc. It may include, but is not limited to. The A may be optionally substituted with one or more substituents selected from the group consisting of oxo, halogen, hydroxy, cyano, amino, C ₁ -C ₆ alkyl, and C ₁ -C ₆ alkoxy.

In some embodiments, R ⁴ can be absent, -A, or -OC(O)-A. In this case, A is pyridinyl, or 2-oxo-1,3-dioxol-4-yl ( ) or 1,3-dioxol-4-yl ( ) can be. The A may be optionally substituted with halogen, hydroxy, cyano, amino, C ₁ -C ₃ alkyl or C ₁ -C ₃ alkoxy. In one embodiment, A may be optionally substituted with C ₁ -C ₃ alkyl, such as methyl or ethyl. In one embodiment, R ⁴ may be absent. In this case, in formula I may be in the methyl ester, ethyl ester or propyl ester structure.

In Formula I of the present invention, X ^- is a monovalent anion. X ^- is not particularly limited as long as it can form a salt with N ⁺ of pyrrolidinium. In one embodiment ^, It may be benzoate or p-toluenesulfonate. For example, X ^- can be chloride, bromide, 4-toluenesulfonate or methanesulfonate. Preferably, X ^- may be bromide.

In one embodiment, the compounds of the invention may be selected from:

Justice

All technical and scientific terms used herein have meanings commonly understood by those skilled in the art and, unless otherwise stated, are based on conventional measurement methods such as pharmacology, pharmaceutical manufacturing, mass spectrometry, NMR, HPLC, biochemistry, etc. , manufacturing methods, and conventional ingredients or materials are used.

Individual features and components of each implementation described and illustrated herein may be combined with features and components of any other implementation without departing from the scope or spirit of the disclosure.

Unless otherwise specified, in this specification and the appended claims, “or” and “and” mean “and/or.” The terms “comprise” and “included” are open-ended and mean that a compound, composition or method may include additional features or ingredients in addition to the specific features or ingredients listed.

In this specification, the numerical range indicated using the term “to” refers to a range that includes the numerical values written before and after the term “to” as the lower limit and upper limit, respectively.

As used herein, the term “optional” or “optionally” means that a subsequently described event or circumstance may or may not occur, and that the description includes instances in which said event or circumstance occurs and instances in which it does not occur. means that For example, the term “optionally substituted” means both substitution and unsubstitution with the specified substituents.

compound

As used herein, unless otherwise stated, the term “alkyl”, whether used alone or as part of a substituent, includes 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms. It refers to saturated straight and branched carbon chains having. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, etc.

The term “alkoxy” refers to the group -O-alkyl. Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, sec-butoxy, iso-butoxy, tert-butoxy, etc.

The term “heteroatom” refers to nitrogen, oxygen or sulfur, and any oxidized form of nitrogen, such as N(O) (N ⁺ -O ^- ) and sulfur, such as S(O) and S(O) ₂ and any quaternized form of the basic nitrogen.

The term “heteroaryl” refers to a heteroaromatic group containing one or more heteroatoms and the remaining ring atom being carbon. The heteroaryl group may include, for example, 1 to 3, 1, or 2 heteroatoms. The heteroaryl group may contain 5 to 10 ring elements, 5 to 7 ring elements, or 5 or 6 ring elements. “Heteroaryl” includes, for example, pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazole, oxazole, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrazolyl. It may be mincedyl, indolyl, benzofuranyl, benzothiophenyl, benzopyrazolyl, benzoimidazolyl, benzooxazolyl, benzoisoxazolyl, benzothiazolyl, or benzoisothiazolyl.

The terms “heterocycloalkyl,” “heterocyclic,” or “heterocycle” refer to a saturated or partially-unsaturated cyclic group containing one or more heteroatoms and the remaining ring atoms being carbon. The heterocycloalkyl group may include, for example, 1 to 3, 1, or 2 heteroatoms. The heterocycloalkyl group may contain 5 to 10 ring elements, 5 to 7 ring elements, or 5 or 6 ring elements. The heterocycloalkyl group is, for example, tetrahydropyranyl, dihydropyranyl, 1,3-dioxolyl, 1,3-dioxolanyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, 2 -Includes, but is not limited to, pyrrolidon-1-yl, morpholinyl, etc.

The term “oxo” refers to the (=O) group.

The term “halogen” as used herein refers to an atom belonging to group 17 of the periodic table. Halogen atoms include fluorine, chlorine, bromine, and iodine, and can be used interchangeably with the term “halo”, which refers to a monovalent functional group composed of halogen.

As used herein, the term “hydroxy” refers to the -OH functional group (hydroxyl group).

The term “cyano” used in this specification is -CN, which refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

As used herein, the term “amino” means -NH ₂ .

As used herein, the term “alkylamino” refers to a group in which one of the two H of the amino group is replaced with an alkyl group. Examples of alkylamino groups include, but are not limited to, methylamino, ethylamino, and propylamino.

As used herein, the term “dialkylamino” means -N(alkyl) ₂ . Here the two alkyls may be the same or different. Examples of dialkylamino substituents include, but are not limited to, dimethylamino, diethylamino, ethylmethylamino, and dipropylamino.

As used herein, the term “nitro” means -NO ₂ .

As used herein, the term “haloalkyl” refers to an alkyl group substituted with one or more halogen atoms. The halogens may be the same (eg, CHF ₂ , -CF ₃ ) or different (eg, CF ₂ Cl). Where otherwise indicated, a haloalkyl group may be optionally substituted with one or more substituents other than halogen. Examples of haloalkyl groups may include, but are not limited to, fluoromethyl, dichloroethyl, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl groups.

As used herein, the term “haloalkoxy” refers to an alkoxy group substituted with one or more halogen atoms, where alkoxy is as defined above. Non-limiting examples of haloalkoxy groups may include fluoromethoxy, dichloroethoxy, trifluoromethoxy, trichloromethoxy, etc.

As used herein, the term “hydroxyalkyl” refers to an alkyl group substituted with one or more -OH groups, where alkyl is as defined above.

As used herein, the term “aminocarbonyl” means amino attached to a carbonyl group through a nitrogen atom.

In this specification " ", "*" or "-" are used to indicate the position at which the substituent is bonded to the remaining residue of the compound. For example, when - is indicated at the end of the substituent, it means that the terminal is bonded to the remaining residue of the compound. In addition, when two or more substituents are connected with "-", it means that the substituent immediately before "-" is bonded to the substitutable atom of the substituent immediately after "-".

The term “solvate” used herein may refer to the compound of the present invention or a salt thereof containing a stoichiometric or non-stoichiometric solvent bound by non-covalent intermolecular forces. Preferred solvents therefor may be volatile, non-toxic, and/or solvents suitable for administration to humans. The solvent may be water, in which case the “solvate” is referred to as “hydrate.”

As used herein, the term “stereoisomer” may refer to a compound of the present invention or a salt thereof that has the same chemical or molecular formula but is optically or sterically different, and specifically includes diastereomers and enantiomers. , or may be geometric isomers.

In some embodiments, the compounds of the invention contain one or more asymmetric centers and may be in the form of a racemate, a single enantiomer, a mixture of enantiomers, a single diastereomer, a mixture of diastereomers, etc. In one embodiment, due to the nature of the asymmetric center or limited rotation, the compounds of the present invention may exist in the form of enantiomers or diastereomers.

When two or more asymmetric centers are present in a compound of the present invention, multiple diastereomers and enantiomers of the chemical structures disclosed herein may exist, including pure isomers, isolated isomers, partially pure isomers, or racemic mixtures. All are intended to fall within the scope of the present invention.

Purification of the isomers and separation of isomer mixtures can be accomplished by standard techniques known in the art. For example, a diastereomeric mixture can be separated into its individual diastereomers by a chromatographic process or crystallization, and a racemate can be separated into its respective enantiomers by a chromatographic process or resolution of the chiral phase.

The compounds of formula I according to the invention are quaternary ammonium salts and can be converted into various salt forms by ion exchange chromatography. The compound may be obtained in the form of a hydrate or solvate. Compounds of formula I can be recovered from the reaction mixture and purified by known methods. When compounds of formula I contain chiral carbons, they can be used as diastereomeric mixtures or as single enantiomers or diastereomers.

The compounds of the present invention can be used in the form of pharmaceutically acceptable salts derived from inorganic acids or organic acids, for example, salts such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malic acid, etc. Lonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methane. It may be a salt derived from sulfonic acid, benzenesulfonic acid, or toluenesulfonic acid.

Pharmaceutically acceptable salts of the compounds are prepared by dissolving the compound of formula (I) in a water-miscible organic solvent, such as acetone, methanol, ethanol, or acetonitrile, adding an excess amount of organic acid or adding an aqueous acid solution of an inorganic acid, and then precipitating it. Alternatively, it can be manufactured by crystallization. Then, the solvent or excess acid in this mixture is evaporated and dried to obtain an addition salt, or the precipitated salt can be prepared by suction filtration.

General manufacturing method of compounds

Compounds according to the present invention can be easily prepared from commercially available starting materials, compounds known in the literature, or intermediates easily prepared therefrom through standard synthetic methods and procedures in the related field.

The methods described herein can be monitored according to any suitable method known in the art. For example, product formation can be determined by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., ¹ H or ¹³ C), infrared spectroscopy, photophotometry (e.g., UV-vis), mass spectrometry, or chromatography. , can be monitored, such as by high performance liquid chromatography (HPLC), gas chromatography (GC), gel-permeation chromatography (GPC), or thin layer chromatography (TLC).

The following general reaction formula generally illustrates a representative method for preparing a compound of formula (I), and those skilled in the art can use starting materials, reaction temperature, reaction conditions suitable for the desired compound based on the manufacturing method specifically disclosed in the Examples herein, The compound of Formula I can be easily prepared by appropriately selecting a catalyst, solvent, processing method, etc. Hereinafter, unless otherwise limited, the designation of each substituent in the formula (I) in the reaction formula is the same as that of the substituent at the corresponding position in the formula (I).

In one aspect, a compound of Formula I can be prepared by reacting intermediate A1 of Formula A1 below with a bromoacetate compound of Formula B1 below.

[Formula A1]

[Formula B1]

BrCH ₂ COO-(CH ₂ ) _p -R ⁴

The bromoacetate compound of Formula B1 can be appropriately selected depending on R ⁴ of the final target, for example, ethyl 2-bromoacetate, methyl 2-bromoacetate, (5-methyl-2-oxo-1,3-di) It may be oxol-4-yl)methyl 2-bromoacetate, etc.

The bromoacetate compound of formula B1 can be changed to other halogenated acetate compounds if necessary. In this case, a halide (eg, I ^- , Cl ^- ) where X ^- is a halogen anion other than bromide can be obtained.

In one aspect, compounds of Formula I can be prepared by reacting intermediate A2 of Formula A2 with a hydroxyalkyl ester compound of Formula B2:

[Formula A2]

[Formula B2]

HO-(CH ₂ ) _p -OC(O)-A

The hydroxyalkyl ester compound of Formula B2 can be appropriately selected depending on R ⁴ of the final target product, and may be, for example, 3-hydroxypropylnicotinate.

Medicinal uses, pharmaceutical compositions and administration methods

Another aspect provides a pharmaceutical composition for preventing or treating muscarinic acetylcholine receptor-mediated diseases, comprising a compound of formula (I), a stereoisomer, hydrate or solvate thereof. The compounds, stereoisomers, hydrates, and solvates of Formula I are as described above.

As used herein, the term "preventing" or "prevention" refers to preventing a disease, e.g., in an individual who may be predisposed to the disease, condition, or disorder but who has not yet experienced or exhibited symptoms or pathology of the disease. , refers to preventing a condition or disorder.

As used herein, the term “treating” or “treatment” refers to inhibiting a disease, e.g., inhibiting a disease, condition or disorder in an individual experiencing or exhibiting pathology or symptoms of the disease, condition or disorder. , preventing further development of pathology and/or symptoms, or improving a disease, e.g., improving a disease, condition or disorder in an individual experiencing or exhibiting pathology or symptoms of the disease, condition or disorder, i.e. , refers to reversing pathology and/or symptoms, such as reducing disease severity.

The muscarinic acetylcholine receptor-mediated disease refers to a disease that can be prevented or treated by inhibiting the binding of acetylcholine to muscarinic receptors, especially muscarinic M3 receptors. The compounds of the present invention have excellent antagonistic activity against muscarinic acetylcholine receptors, especially muscarinic M3 receptors. Specifically, the compound of the present invention was confirmed to have an excellent IC ₅₀ value at the nM level when measuring its antagonistic activity against the human muscarinic M3 receptor according to the method described in Experimental Example 1 herein.

Therefore, the compound of the present invention has excellent antagonistic activity against muscarinic acetylcholine receptors, especially muscarinic M3 receptors, and is useful for the prevention or treatment of various muscarinic acetylcholine receptor-mediated diseases. In some embodiments, the compounds of the present invention can be useful for the prevention or treatment of hyperhidrosis, hypersalivation, chronic obstructive pulmonary disease, chronic bronchitis, asthma, rhinitis, urinary incontinence, overactive bladder, reflux esophagitis, or irritable bowel syndrome.

In one embodiment, the pharmaceutical composition may include conventional pharmaceutically acceptable carriers, excipients, or additives. The pharmaceutical composition can be formulated according to conventional methods, and can be administered in various oral dosage forms such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, and parenteral dosage forms such as intramuscular, intravenous, or subcutaneous administration. , or may be prepared in a form for topical application to the skin. The pharmaceutical composition may be a single composition or separate compositions. The pharmaceutical composition includes a compound, stereoisomer, hydrate or solvate according to one aspect as an active ingredient of the pharmaceutical composition.

When the pharmaceutical composition is prepared in the form of an oral dosage form, examples of additives or carriers used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, and stearic acid. Examples include calcium, gelatin, talc, surfactants, suspending agents, emulsifiers, and diluents. When the pharmaceutical composition of the present invention is prepared in the form of an injection, the additives or carriers include water, saline solution, aqueous glucose solution, similar aqueous sugar solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester. , glycerides, surfactants, suspending agents, emulsifiers, etc.

The dosage of the pharmaceutical composition is an amount effective for treatment or prevention of an individual or patient, and can be administered orally or parenterally depending on the purpose. When administered orally, the dosage is 0.01 to 1000 mg per kg of body weight per day, more specifically, 0.1 to 300 mg per day based on the active ingredient, and when administered parenterally, the dosage is administered per kg of body weight per day based on the active ingredient. It can be administered once to several times in doses of 0.01 to 100 mg, more specifically, 0.1 to 50 mg. The administered dose for a specific individual or patient must be determined in light of various related factors such as the patient's weight, age, gender, health status, diet, administration time, administration method, and severity of the disease, and can be adjusted appropriately by an expert. It should be understood that the above dosage is not intended to limit the scope of the present invention in any way. A physician or veterinarian having ordinary skill in the relevant art can easily determine and prescribe an effective amount of the required pharmaceutical composition. For example, a physician or veterinarian may start the dose of a compound of the invention used in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. can be increased.

In one embodiment, the pharmaceutical composition includes within its scope a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of at least one of the compounds according to one embodiment, alone or in combination with a pharmaceutical carrier. The term “therapeutically effective amount” or “effective amount” means an amount sufficient to produce a beneficial or desired clinical outcome, e.g., an amount sufficient to alleviate, ameliorate, stabilize, reverse, slow or delay the progression of a disease.

When the composition of the present invention is used for the prevention or treatment of hyperhidrosis, the composition of the present invention can be applied topically to the skin as a solid, semi-solid, powder, gel, ointment, cream, lotion, foam, solution, suspension, aerosol, patch, emulsion, etc. It can be formulated into an application preparation. In one embodiment, topical application formulations of the invention may be gels, creams, emulsions, lotions, or sprays.

In one embodiment, gels, ointments and creams may be formulated using, for example, suitable thickening and/or gelling agents and/or aqueous or oily bases. For example, water and/or oils such as liquid paraffin, or vegetable oils such as arachis oil or castor oil, or glycol solvents such as propylene glycol or 1,3-butanediol. For example, thickeners include soft paraffin, aluminum stearate, cetostearyl alcohol, polyethylene glycol, wool fat, hydrogenated lanolin and beeswax and/or glyceryl monostearate and/or nonionic emulsifiers. In one embodiment, lotions may be formulated using an aqueous or oily base and may also include one or more of emulsifiers, dispersants, suspending agents, thickening agents, solvents, colorants, and flavoring agents. The powder may be formed with the aid of any suitable powder base, for example talc, lactose or starch. Spray compositions can, for example, be formulated as aerosols using a suitable propellant, such as dichlorodifluoromethane or trichlorofluoromethane.

The proportions of active ingredients in topical application formulations according to the invention will vary depending on the compound used, the type of formulation, and the specific condition for which the composition is to be administered. For example, the topically applied formulation may comprise from about 0.1% to about 50%, from about 1% to about 30%, or from about 5% to about 20% of the compound of the invention.

When used to treat hyperhidrosis, the topical application formulation can be applied topically as needed to the area of the subject's skin in need of sweating reduction, such as the subject's palms, soles, groin, armpits, or face. The topically applied preparation may be applied to the subject as needed, for example, more than once a week, 3 to 4 times a week, once a day, twice a day, or three times a day.

Another aspect provides a method of preventing or treating a muscarinic acetylcholine receptor mediated disease comprising administering or topically applying to a subject a compound of Formula (I), a stereoisomer, hydrate or solvate thereof.

Among the terms or elements mentioned in the description of the method, those that are the same as those mentioned above are the same as those mentioned above.

The administration may be oral administration or parenteral administration. When administered orally, the dosage is 0.01 to 1000 mg per kg of body weight per day, more specifically, 0.1 to 300 mg per day based on the active ingredient, and when administered parenterally, the dosage is administered per kg of body weight per day based on the active ingredient. It can be administered once to several times in doses of 0.01 to 100 mg, more specifically, 0.1 to 50 mg.

The administered dose for a specific individual or patient must be determined in light of various related factors such as the patient's weight, age, gender, health status, diet, administration time, administration method, and severity of the disease, and can be adjusted appropriately by an expert. there is.

In some embodiments, when the methods of the invention are performed to treat hyperhidrosis in a subject, the formulations of the invention may be applied topically to areas of the subject's skin in need of reduced sweating. In one embodiment, the subject's skin area may include the subject's palm, sole, groin, armpit, or face. The compounds of the present invention can be applied topically to the subject as needed, for example, once a week or more, 3 to 4 times a week, once a day, twice a day, or three times a day. Depending on the method of the present invention, sweating can be reduced by about 10% to about 99%, about 30% to about 80%, or at least 50%.

As used herein, the term "individual" refers to a subject in need of treatment for a disease, and more specifically, mammals such as humans or non-human primates, mice, dogs, cats, horses, and cattle. means.

Another aspect is the medicinal use of a compound of formula (I), a stereoisomer, hydrate or solvate thereof, for the prevention or treatment of muscarinic acetylcholine receptor mediated diseases; or the use of a compound of formula (I), a stereoisomer, hydrate or solvate thereof for preparing a medicament for preventing or treating muscarinic acetylcholine receptor-mediated diseases. Among the terms or elements mentioned in the description of the above use, those that are the same as those already mentioned are as described above.

Compounds of formula (I), their stereoisomers, hydrates or solvates exhibit excellent muscarinic M3 receptor antagonistic activity, which can be used to treat hyperhidrosis, hypersalivation, chronic obstructive pulmonary disease, asthma, irritable bowel syndrome, urinary incontinence, rhinitis, glaucoma, cardiac arrhythmias, etc. It is useful for preventing or treating various diseases, especially hyperhidrosis.

Figure 1 shows the number of sweating spots per hindfoot in the normal group (vehicle), the hyperhidrosis-induced group, the positive control group administered 20% sophironium, and the experimental group administered 20% Compound 1 and 20% Compound 3, respectively. This is a graph showing the results. ## p<0.01 vs normal group; ** p<0.01 vs induced group (one-way ANOVA, LSD post hoc test)

This will be described in more detail through examples below. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Manufacturing example

Preparation Example 1-1: (R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carba synthesis of mates

3'-Chloro-4'-fluoro-[1,1'-biphenyl]-2-carboxylic acid (0.80 g, 3.19 mmol) and (R)-(1-methylpyrrolidin-3-yl) Methanol (0.40 g, 3.51 mmol) was dissolved in toluene (8 mL), triethylamine (0.97 g, 9.57 mmol) and diphenylphosphorylazide (1.32 g, 4.79 mmol) were added at room temperature, and then placed outside under nitrogen atmosphere. It was stirred at a temperature of 120°C for 12 hours. After completion of the reaction, the temperature of the reactant was cooled to room temperature and then 1N hydrochloric acid solution (30 mL) was added. This mixed solution was extracted with ethyl acetate and washed with sodium bicarbonate. The organic layer was dehydrated with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain the title compound (0.50 g, 43%).

MS: m/z [MH] ^- = 362.8

¹ HNMR (400 MHz, CDCl3): δ 7.89 (d, J = 7.6 Hz, 1H), 7.44-7.39 (m, 1H), 7.38-7.33 (m, 1H), 7.25-7.12 (m, 4H), 6.97 (t, J = 6.8 Hz, 1H), 4.11-3.97 (m, 2H), 2.92-2.85 (m, 1H), 2.83-2.58 (m, 4H), 2.50 (s, 3H), 2.10-2.03 (m , 1H), 1.71-1.63 (m, 1H)

Preparation Example 1-2: (R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carba Synthesis of mates (alternative method)

Step 1: Synthesis of 3’-chloro-4’fluoro-[1,1’-biphenyl]-2-amine

(3-chloro-4-fluoro)phenylboronic acid (5.0 g, 28.7 mmol), sodium carbonate (6.6 g, 63.8 mmol) and tetrakistriphenylphosphinepalladium (1.8 g, 1.56 mmol) were added to the reactor in toluene (40 mL) and water (20 mL), then 2-bromo-4-fluoroaniline (3.8 g, 22.1 mmol) was added and stirred for 18 hours at an external temperature of 95 degrees. After completion of the reaction, saturated ammonium solution (50 mL) and dichloromethane (50 mL) were added, stirred, and the aqueous layer and organic layer were separated. The aqueous layer was extracted with dichloromethane (2 .

Step 2: Synthesis of (R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate

Triphosgene (1.2 g, 4.0 mmol) was dissolved in dichloromethane (30 mL) and then 3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-amine (2.5 g, 11.3 mmol). and triethylamine (3.4 g, 33.9 mmol) were added at 0°C. After reaction at 0°C for 1 hour, (R)-(1-methylpyrrolidin-3-yl)methanol (1.2 g, 10.7 mmol) was added dropwise and stirred at room temperature for 1 hour. After completion of the reaction, the extract was extracted using water and dichloromethane, dehydrated with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain the title compound (1.2 g, 31%).

MS: m/z [M+H] ⁺ = 363.2

Preparation Example 2: (S)-(1-methylpyrrolidin-2-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate synthesis

As a starting material, 3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-carboxylic acid was reacted with (S)-(1-methylpyrrolidin-2-yl)methanol. The title compound (0.50 g, 43%) was obtained in the same manner as Preparation Example 1-1.

MS: m/z [MH] ^- = 362.8

^1H NMR (400 MHz, DMSO-d6): δ 8.06 (d, J = 7.6 Hz, 1H), 7.47-7.36 (m, 2H), 7.30-7.22 (m, 2H), 7.21-7.11 (m, 2H) ), 6.56 (s, 1H), 4.27-4.08 (m, 2H), 3.13-3.07 (m, 1H), 2.56-2.46 (m, 1H), 2.42 (s, 3H), 2.32-2.22 (m, 1H) ), 1.99-1.59 (m, 4H)

Preparation Example 3: Synthesis of 2-(1-methylpyrrolidin-2-yl)ethyl(3’-chloro-4’-fluoro-[1,1’-biphenyl]-2-yl)carbamate

As a starting material, 3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-carboxylic acid is mixed with 2-(1-methylpyrrolidin-2-yl)ethan-1-ol. The reaction was performed in the same manner as Preparation Example 1-1 to obtain the title compound (0.67 g, 45%).

MS: m/z [MH] ^- = 376.8

^1H NMR (400 MHz, CDCl3): δ 7.84 (d, J = 7.6 Hz, 1H), 7.58-7.38 (m, 1H), 7.32-7.28 (m, 1H), 7.24-7.19 (m 1H), 7.14 -7.03 (m, 1H), 6.95 (t, J = 6.4 Hz, 2H), 6.88 (s, 1H), 4.17-4.00 (m, 2H), 3.55-3.51 (m, 1H), 2.79-2.49 (m , 5H), 2.16-2.05 (m, 2H), 1.95-1.54 (m, 4H)

Preparation Example 4: Synthesis of 2-(1-methylpyrrolidin-2-yl)ethyl[1,1’-biphenyl]-2-ylcarbamate

2-Isocyanato-1,1'-biphenyl (0.50 g, 2.96 mmol) was dissolved in tetrahydrofuran (10 mL) and then 2-(1-methylpyrrolidin-2-yl)ethanol (0.38 g) , 2.96 mmol) was added dropwise. The reaction was stirred at room temperature for 14 hours to complete the reaction, concentrated under reduced pressure, and then purified by column chromatography to obtain the title compound (0.70 g, 72%).

MS: m/z [M+H] ⁺ = 325.3

Preparation Example 5: Synthesis of (S)-(1-methylpyrrolidin-2-yl)methyl[1,1’-biphenyl]-2-ylcarbamate

The title compound ( 0.40 g, 84%) was obtained.

MS: m/z [M+H] ⁺ = 311.1

Preparation Example 6: Synthesis of (R)-(1-methylpyrrolidin-2-yl)methyl[1,1’-biphenyl]-2-ylcarbamate

The title compound ( 0.45 g, 94%) was obtained.

MS: m/z [M+H] ⁺ = 311.0

Preparation Example 7: Synthesis of 2-(1-methylpyrrolidin-2-yl)ethyl(5-fluoro-[1,1’-biphenyl]-2-yl)carbamate

Step 1: Synthesis of 5-fluoro-[1,1'-biphenyl]-2-amine

Phenylboronic acid (3.9 g, 31.9 mmol), potassium carbonate (8.8 g, 63.8 mmol), and tetrakistriphenylphosphinepalladium (1.8 g, 1.56 mmol) were added to the reactor in toluene (40 mL), water (20 mL), After dissolving in ethanol (10 mL) mixed solvent, 2-bromo-4-fluoroaniline (3.0 g, 15.9 mmol) was added and stirred at an external temperature of 95°C for 16 hours. After completion of the reaction, saturated ammonium solution (50 mL) and dichloromethane (50 mL) were added, stirred, and the aqueous layer and organic layer were separated. The aqueous layer was extracted with dichloromethane (2 * 50 mL), the organic layer was collected, dehydrated with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain the title compound (2.9 g, 97%). .

Step 2: Synthesis of 2-(1-methylpyrrolidin-2-yl)ethyl(5-fluoro-[1,1'-biphenyl]-2-yl)carbamate

As a starting material, 5-fluoro-[1,1'-biphenyl]-2-amine was reacted with 2-(1-methylpyrrolidin-2-yl)ethanol to produce the same reaction as step 2 of Preparation Example 1-2. The title compound (2.15 g, 50%) was obtained by this method.

MS: m/z [M+H] ⁺ = 343.2

Preparation Example 8: Synthesis of (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl 2-bromoacetate

4-(Hydroxymethyl)-5-methyl-1,3-dioxol-2-one (1.0 g, 7.7 mmol) was dissolved in dichloromethane (10 mL) and then 2-bromoacetyl bromide (1.5 g) , 7.7 mmol) and pyridine (1.8 g, 23.1 mmol) were added dropwise at 0°C. The reaction mixture was stirred at room temperature for 18 hours and then extracted with saturated ammonium solution and dichloromethane. The organic layer was dehydrated with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by column chromatography to obtain the title compound (1.0 g, 53%).

Preparation Example 9: (3R)-1-(carboxymethyl)-3-((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl-carbamoyl)oxy ) Synthesis of methyl)-1-methylpyrrolidin-1-ium bromide

(3R)-3-((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl-carbamoyl)oxy)methyl)-1-(2- Ethoxy-2-oxomethyl)1-methylpyrrolidin-1-ium bromide (0.30 g, 0.69 mmol) was dissolved in methanol (10 mL), and then sodium hydroxide solution (0.69 mL, 2N) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature for 2 hours and concentrated under reduced pressure. The concentrated residue was purified by prep-HPLC with acetonitrile and water containing 0.1% hydrogen bromide to obtain the title compound (0.20 g, 69%). .

Example

Example 1: (3R)-3-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamoyl)oxy)methyl)-1-( 2-Ethoxy-2-oxoethyl)-1-methylpyrrolidin-1-ium bromide (Compound 1)

(R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.50 g, 1.38 mmol) was dissolved in acetonitrile (10 mL), and then ethyl 2-bromoacetate (0.46 g, 2.76 mmol) was added dropwise. After the reaction was completed by stirring for 12 hours under nitrogen atmosphere at 25°C, the reaction mixture was concentrated under reduced pressure. The concentrated residue was purified by column chromatography to obtain compound 1 (0.38 g, 52%).

MS: m/z [M-Br] ^- = 449.2

1H NMR (400 MHz, DMSO-d6): δ 9.00 (d, J = 5.6 Hz, 1H), 7.57 (d, J = 6.4 Hz, 1H), 7.52-7.46 (m, 1H), 7.41-7.33 (m , 3H), 7.31-6.95 (m, 2H), 4.56 (d, J= 21.2 Hz, 2H), 4.29-4.18 (m, 2H), 4.14-3.42 (m, 6H), 3.23-3.19 (m, 3H) ), 2.88-2.81 (m, 1H), 2.28-2.21 (m, 1H), 1.90-1.75 (m, 1H), 1.25 (t, J = 6.8 Hz, 3H)

Example 2: 2-(2-(([1,1'-biphenyl]-2-ylcarbamoyl)oxy)ethyl)-1-(2-ethoxy-2-oxoethyl)-1-methylp Rolidine-1-ium bromide (Compound 2)

Compound 2 ( 0.33 g, 65%) was obtained.

MS: m/z [M-Br] ^- = 411.3

^1H NMR (400 MHz, DMSO-d6): δ 8.70 (s, 1H), 7.50-7.27 (m, 9H), 4.67-4.27 (m, 1H), 4.26-4.18 (m, 3H), 4.11-3.83 (m, 3H), 3.77-3.46 (m, 2H), 3.10 (d, J = 112.4 Hz, 3H), 2.32-2.13 (m, 2H), 2.08-1.99 (m, 2H), 1.92-1.69 (m , 2H), 1.25 (t, J = 7.2 Hz, 3H)

Example 3: (2S)-2-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamoyl)oxy)methyl)-1-( 2-Ethoxy-2-oxoethyl)-1-methylpyrrolidin-1-ium bromide (Compound 3)

(S)-(1-methylpyrrolidin-2-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.40 g, 3.51 Compound 3 (0.40 g, 69%) was obtained in the same manner as in Example 1 using mmol).

MS: m/z [M-Br] ^- = 448.0

^1H NMR (400 MHz, DMSO-d6): δ 7.63-7.57 (m, 1H), 7.50-7.29 (m, 6H), 4.48-4.31 (m, 3H), 4.26-4.09 (m, 3H), 3.95 -3.65 (m, 2H), 3.43-3.41 (m, 2H), 3.09 (s, 2H), 2.30-1.99 (m, 3H), 1.95-1.82 (m, 1H), 1.27-1.22 (m, 3H)

Example 4: 1-(2-ethoxy-2-oxoethyl)-2-(2-(((5-fluoro-[1,1'-biphenyl]-2-yl)carbamoyl)oxy) Ethyl)-1-methylpyrrolidin-1-ium bromide (Compound 4)

Example 1 using 2-(1-methylpyrrolidin-2-yl)ethyl(5-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.50 g, 1.46 mmol) Compound 4 (0.53 g, 70%) was obtained in the same manner as above.

MS: m/z [M-Br] ^- = 429.2

¹ H NMR (400 MHz, CDCl3): δ 7.84 (dd, J = 15.6, 10.4 Hz, 1H), 7.55-7.30 (m, 5H), 7.19-6.79 (m, 3H), 5.40-4.80 (m, 1H) ), 4.68 (dd, J = 20.0, 17.2 Hz, 1H), 4.43 (m, 1H), 4.33-4.05 (m, 6H), 3.60 (s, 2H), 3.13 (s, 1H), 2.60-1.78 ( m, 6H), 1.29 (td, J = 7.2, 1.6 Hz, 3H)

Example 5: (2S)-2-((([1,1'-biphenyl]-2-ylcarbamoyl)oxy)methyl)-1-(2-ethoxy-2-oxoethyl)-1- Methylpyrrolidin-1-ium bromide (Compound 5)

Compound was prepared in the same manner as in Example 1 using (S)-(1-methylpyrrolidin-2-yl)methyl[1,1'-biphenyl]-2-ylcarbamate (0.32 g, 1.93 mmol). 5 (0.35 g, 57%) was obtained.

MS: m/z [M-Br] ^- = 397.3

¹ H NMR (400 MHz, DMSO-d6): δ 9.06-8.94 (m, 1H), 7.65-7.12 (m, 9H), 4.69-4.04 (m, 7H), 3.96-3.57 (m, 2H), 3.28 -2.98 (m, 3H), 2.36-1.72 (m, 4H), 1.28-1.22 (m, 3H)

Example 6: (3R)-3-((([1,1'-biphenyl]-2-ylcarbamoyl)oxy)methyl)-1-(2-ethoxy-2-oxoethyl)-1- Methylpyrrolidin-1-ium bromide (Compound 6)

Compound was prepared in the same manner as in Example 1 using (R)-(1-methylpyrrolidin-2-yl)methyl[1,1'-biphenyl]-2-ylcarbamate (0.45 g, 1.45 mmol). 6 (0.30 g, 52%) was obtained.

MS: m/z [M-Br] ^- = 397.2

^1H NMR (400 MHz, CDCl3) δ 8.81-8.78 (m, 1H), 7.46-7.33 (m, 9H), 4.52-4.49 (m, 2H), 4.26-4.21 (m, 2H), 4.02-3.98 ( m, 2H), 3.88-3.61 (m, 3H), 3.31-3.26 (m, 1H), 3.19-3.17 (m, 3H), 2.85-2.78 (m, 1H), 2.28-2.14 (m, 1H), 1.94-1.74 (m, 1H), 1.27-1.24 (m, 3H).

Example 7: 2-(2-(((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamoyl)oxy)ethyl)-1-(2 Toxy-2-oxoethyl)-1-methylpyrrolidin-1-ium bromide (Compound 7)

2-(1-methylpyrrolidin-2-yl)ethyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.67 g, 1.78 mmol) Compound 7 (0.45 g, 47%) was obtained in the same manner as in Example 1.

MS: m/z [M-Br] ^- = 463.2

¹ H NMR (400 MHz, DMSO-d6): δ 8.94 (s, 1H), 7.58 (d, J = 6.8 Hz, 1H), 7.51-7.30 (m, 6H), 4.68-4.31 (m, 1H), 4.30-4.20 (m, 3H), 4.15-3.90 (m, 3H), 3.75-3.64 (m, 1H), 3.63-3.51 (m, 1H), 3.26-2.97 (m, 3H), 2.28-2.20 (m) , 2H), 2.08-2.00 (m, 2H), 1.93-1.75 (m, 2H), 1.25 (t, J = 6.8 Hz, 3H).

Example 8: (3R)-3-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamoyl)oxy)methyl)-1-( 2-methoxy-2-oxoethyl)-1-methylpyrrolidin-1-ium bromide (Compound 8)

(R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.30 g, 0.83 mmol) was dissolved in acetonitrile (10 mL), and then methyl 2-bromoacetate (0.19 g, 1.25 mmol) was added dropwise. After the reaction was completed by stirring for 12 hours under nitrogen atmosphere at 25°C, the reaction mixture was concentrated under reduced pressure. The concentrated residue was purified by column chromatography to obtain compound 8 (66 mg, 16%).

MS: m/z [M-Br] ^- = 435.2

^1H NMR (400 MHz, DMSO): δ 8.99 (d, J = 6.0 Hz, 1H), 7.58 (dd, J = 7.2, 1.2 Hz, 1H), 7.52-7.46 (m, 1H), 7.44-7.30 ( m, 5H), 4.58 (d, J = 21.6 Hz, 2H), 4.15-3.82 (m, 3H), 3.76-3.59 (m, 2H), 3.48-3.35 (m, 1H), 3.27-3.12 (m, 3H), 2.99-2.78 (m, 1H), 2.37-2.11 (m, 1H), 2.01-1.66 (m, 1H)

Example 9: (3R)-3-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]2-yl)carbamoyl)oxy)methyl)-1-methyl- 1-(2-((5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy)2-oxoethyl)pyrrolidin-1-ium bromide (Compound 9)

(R)-(1-methylpyrrolidin-3-yl)methyl(3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl)carbamate (0.30 g, 0.83 mmol) was dissolved in acetonitrile (10 mL), and then (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl 2-bromoacetate (0.31 g, 1.25 mmol) was added dropwise. After the reaction was completed by stirring for 12 hours under nitrogen atmosphere at 25°C, the reaction mixture was concentrated under reduced pressure. The concentrated residue was purified by prep-HPLC with acetonitrile and water containing 0.1% hydrogen bromide to obtain compound 9 (104 mg, 20%).

MS: m/z [M-Br] ^- = 533.2

^1H NMR (400 MHz, DMSO): δ 8.96 (d, J = 8.4 Hz, 1H), 7.84-7.16 (m, 7H), 5.14 (s, 2H), 4.57 (d, J = 15.6 Hz, 2H) , 4.17-3.56 (m, 5H), 3.45-3.34 (m, 1H), 3.26-3.11 (m, 3H), 2.90-2.73 (m, 1H), 2.34-2.12 (m, 4H), 1.98-1.78 ( m, 1H)

Example 10: (3R)-3-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]2-yl)carbamoyl)oxy)methyl)-1-methyl- 1-(1-(2-(3-(nicotinoyloxy)propoxy)-2-oxoethyl)pyrrolidin-1-ium bromide (Compound 10)

(3R)-1-(carboxymethyl)-3-(((((3'-chloro-4'-fluoro-[1,1'-biphenyl]-2-yl-carbamoyl)oxy)methyl)- 1-Methylpyrrolidin-1-ium bromide (0.20 g, 0.48 mmol), 1-hydroxybenzotriazole (71 mg, 0.53 mmol), 3-hydroxypropylnicotinate (96 mg, 0.53 mmol) After dissolving in dichloromethane (20 mL), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (101 mg, 0.53 mmol) was added and stirred at room temperature for 12 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the concentrated residue was purified by prep-HPLC with acetonitrile and water containing 0.1% hydrogen bromide to obtain compound 10 (50 mg, 16%).

MS: m/z [M-Br] ^- = 583.8

¹ H NMR (400 MHz, DMSO): δ 9.20-9.09 (m, 1H), 9.01-8.90 (m, 1H), 8.87-8.79 (m, 1H), 8.51-8.32 (m, 1H), 7.68-7.60 (m, 1H), 7.57-7.50 (m, 1H), 7.50-7.42 (m, 1H), 7.41-7.24 (m, 5H), 4.75-4.45 (m, 3H), 4.45-4.29 (m, 5H) , 4.11-3.83 (m, 8H), 3.38-3.27 (m, 2H), 3.24-3.08 (m, 3H), 2.90-2.76 (m, 1H), 2.32-2.04 (m, 3H), 1.92-1.76 ( m, 1H).

Experiment example

Experimental Example 1: In vitro muscarinic M3 receptor antagonism

1. Human muscarinic M3 receptor binding ability test

The affinity (Ki) of the compounds of the invention for the human muscarinic acetylcholine M3 receptor was determined by a competitive filtration binding assay using the radio-labeled M3 receptor antagonist, [3H]-methylscopolamine.

Cell membrane protein derived from CHO-K1 overexpressing human muscarinic M3 receptor (PerkinElmer), [3H]-methyl scopolamine, and various concentrations of test substances were incubated in 0.1 mL of Tris-HCl buffer at 25°C. The culture was incubated for 120 minutes, suction filtered through a glass filter (Whatman GF/B), and the filter was washed six times with 0.5 mL of cold Tris-HCl buffer. The radioactivity of [3H]-methylscopolamine adsorbed on the filter was measured by adding 50 uL of microscint 20 (Packard) to each well, incubating in an orbital shaker for 15 minutes, and using TopCount ^TM . Non-specific binding was evaluated in the presence of N-methylscopolamine at a 200- to 300-fold concentration. To evaluate the binding ability of the example compounds to the muscarinic M3 receptor, the method of Cheng and Prusoff, Biochem. Pharmacol., 22, 3099, 1973], the dissociation constant (Ki) was calculated from the concentration (IC ₅₀ ) of the test substance that inhibits the binding of the labeling ligand, [3H]-methylscopolamine, by 50%. The lower the dissociation constant (Ki), the stronger the compound's binding ability to the human muscarinic M3 receptor.

The M3 receptor binding ability of each test substance is shown in Table 1 below.

Binding capacity to human muscarinic M3 receptor compound IC ₅₀ (nM) M3 receptor binding capacity Ki (nM) Compound 1 46.6 5.17 Compound 3 100 11.1 Compound 5 173 19.3

As can be seen from Table 1, the compounds of the present invention were confirmed to have excellent IC ₅₀ values at the nM level for the human muscarinic M3 receptor.

2. Human muscarinic M3 receptor antagonism test

Antagonism against the M3 receptor was evaluated using CHO-K1 cells (CHO-K1 mt aequorin, PerkinElmer) transfected with the human muscarinic M3 receptor.

Recombinant cells grown 18 hours before testing in antibiotic-free medium were separated by gentle flushing with PBS-EDTA (5mM EDTA), recovered by centrifugation, and incubated in buffer (DMEM/HAM's F12 containing HEPES + 0.1% BSA protease). It was resuspended. Cells were incubated with Coelenterazine h (Molecular Probes) at room temperature for at least 4 hours. A dose response curve was obtained using a reference compound (4-DAMP) prior to compound evaluation.

For agonist evaluation, 50 μL of cell suspension was injected into 50 μL of test or reference compound plated in a 96-well plate. Luminescence was recorded using a Hamamatsu Functional Drug Screening System 6000 (FDSS 6000).

After incubation for 15 minutes after the first injection, 100 μL of the reference agonist (acetylcholine) at a concentration corresponding to EC ₈₀ was injected into 100 μL of the mixture of cell suspension and test compound for antagonist testing. Luminescence was recorded using FDSS6000.

The antagonist activity of the test compound was calculated as the percentage inhibition of the reference activity at the reference agonist (acetylcholine) EC ₈₀ concentration, and then the IC ₅₀ value was calculated. The M3 receptor antagonistic ability for each test substance is shown in the table below.

Human muscarinic M3 receptor antagonistic activity compound IC ₅₀ (nM) Compound 1 49.3 Compound 2 221 Compound 3 91.7 Compound 4 >250 Compound 5 46.8 Compound 6 152 Compound 7 160

Experimental Example 2: Evaluation of drug efficacy in pilocarpine-induced mouse hyperhidrosis model

An iodine-starch sweat test was performed to confirm the antiperspirant efficacy of the test substance in the pilocarpine-induced hyperhidrosis model.

Six-week-old male ICR mice (Orient Bio) were purchased, acclimatized for one week, and then grouped using body weight. The test group is shown in Table 3 below.

Test group composition test group Pilocarpine
5 mg/kg, i.p. Dosage
(%) Administration volume
(μL/hind footpad) population Normal group (vehicle) - - 10 8 Inducer group (vehicle) + - 10 8 Positive control group (Sofpironium) + 20 10 8 Compound 1 + 20 10 8 Compound 3 + 20 10 8

Vehicle: absolute ethanol

Four hours before pilocarpine administration, the test substance was prepared at 20% in absolute ethanol (Daejung) and applied at 10 μL to the hind footpads of both sides of the mouse. Four hours after application of the test substance, an anesthetic (rompum:ketamine = 1:4) was administered intraperitoneally, and then pilocarpine (5 mg/kg/5 mL) was administered into the abdominal cavity of the anesthetized animal. Iodine (Sigma-Aldrich) was dissolved in ethanol at 3.5% and applied to the right and left hindlimbs using a brush. After drying for about 1 minute, starch (Milipore) was suspended in castor oil (Sigma-Aldrich) at a concentration of 10% and applied to the same location using a brush. Ten minutes after pilocarpine administration, photos of the plantar skin were taken and sweating spots/foodpad per plantar of each subject were analyzed. Sopyrronium bromide (JHCHEM) was used as a positive control.

All data obtained from the experiment are expressed as mean ± standard error, and all results were analyzed by one-way ANOVA using SPSS (version 20, IBM SPSS Statistics, USA), and significance was verified by post hoc test using LSD test. .

The test results are shown in Table 4 below.

Number of sweating spots on the left and right hindfoot plantar test group Number of sweating spots (mean ± standard error) Normal group (vehicle) 17.00 ± 4.21 Induced group (vehicle) 116.88 ± 17.51 ^## Positive control (20% sophironium) 42.13 ± 8.35** 20% compound 1 32.25 ± 6.14** 20% compound 3 37.50 ± 13.02**

## p<0.01 vs normal group; ** p<0.01 vs induced group (one-way ANOVA, LSD post hoc test)

The number of sweating spots was 17.00 ± 11.90 in the normal group (vehicle) not administered pilocarpine, and 116.88 ± 49.54 in the sweating-induced group administered pilocarpine, confirming the induction of hyperhidrosis (p<0.01).

The number of sweating spots in the Compound 1 and Compound 3 application groups was 32.25 ± 6.14 and 37.50 ± 13.02, respectively, which was significantly reduced compared to the sweating-inducing group (p<0.01; see Table 4 and Figure 1).

In addition, the number of sweating spots of the positive control (sophironium) at the same concentration was confirmed to be 42.13 ± 8.35, confirming that the compounds of the present invention exhibit an equally excellent antiperspirant ability compared to the positive control (sophironium).

Although the present invention has been described herein based on exemplary embodiments, the present invention is not limited to the described embodiments, and all changes, modifications, modifications and alternatives that do not depart from the spirit and essential characteristics of the present invention are within the scope of the present invention. It should be understood as being included in .

Claims (11)

Compounds represented by the following formula (I), stereoisomers, hydrates or solvates thereof:
Formula I

R ¹ and R ² are each independently hydrogen or halogen;
R ³ is hydrogen or C ₁ -C ₆ alkyl;
R ⁴ is H, -A or -OC(O)-A;
A is pyridinyl or 2-oxo-1,3-dioxol-4-yl, wherein the pyridinyl or 2-oxo-1,3-dioxol-4-yl is optionally substituted with C ₁ -C ₆ alkyl. can;
n and m are each independently integers from 0 to 3;
k is an integer from 1 to 3,
p is an integer from 1 to 5,
X ^- is a monovalent anion. According to claim 1,
n and m are each independently 0, 1 or 2. The compound, stereoisomer, hydrate or solvate thereof. According to claim 1,
R ³ is C ₁ -C ₃ alkyl, a compound, a stereoisomer, hydrate or solvate thereof. According to claim 1,
p is an integer from 1 to 3. A compound, stereoisomer, hydrate or solvate thereof. According to claim 1,
X ^-silk chloride, bromide, yiodide, sulfate, phosphate, methane sulfonate, knitrate, maleate, acetate, citrate, fumarate, tart, oxalate A compound selected from the group consisting of toluenesulfonates, a stereoisomer, hydrate or solvate thereof. According to claim 5,
X ^- is a bromide, a compound, a stereoisomer, hydrate or solvate thereof. According to claim 1,
Compounds selected from the following group, stereoisomers, hydrates or solvates thereof:

A pharmaceutical composition for preventing or treating muscarinic acetylcholine receptor-mediated diseases, comprising the compound of any one of claims 1 to 7, a stereoisomer, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, A pharmaceutical composition, wherein the muscarinic acetylcholine receptor-mediated disease is selected from the group consisting of hyperhidrosis, hypersalivation, chronic obstructive pulmonary disease, chronic bronchitis, asthma, rhinitis, urinary incontinence, overactive bladder, reflux esophagitis, and irritable bowel syndrome. According to claim 8,
A pharmaceutical composition wherein the muscarinic acetylcholine receptor-mediated disease is hyperhidrosis.
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