Compounds having O-GlcNAcase inhibitory activity and uses thereof

Abstract

The present invention provides compounds that have O-GlcNAcase inhibitory activity and can reduce tau phosphorylation in vivo, and pharmaceutical compositions containing the same for treating or preventing diseases caused by tau hyperphosphorylation.

Description

Technical Field

[0001] The present invention relates to a novel compound having O-GlcNAcase inhibitory activity and its use.

Background Art

[0002] Microtubule-associated protein tau (MAPT) is an essentially unstructured protein that is refined through various reactions such as glycosylation and phosphorylation. The Tau protein aggregates by sequential hyperphosphorylation to form paired helical filaments (PHFs), which then form neurofibrillary tangles (NFTs). The NFTs thus formed are one of the major pathological features of Alzheimer's disease (AD) and a wider range of neurodegenerative diseases called tauopathies. The normal function of phosphorylated tau is to promote and stabilize microtubule assembly. In healthy individuals, the average contains 1.9 groups of phosphate groups per tau protein molecule. In the brains of pathologically confirmed AD patients, soluble tau has an average of 2.6 phosphate groups per molecule, and tau purified from PHFs has an average of 6 to 8 phosphate groups per molecule. In AD patients, the degree of phosphorylation is increased 3 to 4 times compared with tau isolated from healthy brain tissue. Tau hyperphosphorylation not only promotes its own aggregation but also prevents the binding of tau to microtubules and reduces microtubule stability.

[0003] According to research conducted 10 years ago, bovine tau is extensively post-translationally modified by O-GlcNAc binding, and such modifications were identically observed in human tau. O-GlcNAc is found from the hydroxyl side chains of serine and threonine residues of diverse nuclear and cytoplasmic proteins, and in some cases, it was revealed to originate from residues of proteins known to have been phosphorylated. Similar to phosphorylation, O-GlcNAcylation is a dynamic modification and can be transferred to and removed from proteins several times during the lifespan of the polypeptide backbone. Such a dynamic cycle of O-GlcNAc is mediated by two enzymes. Uridine diphosphate N-acetyl-D-glucosamine polypeptidyltransferase OGT targets the hudroxyl group of the receptor protein and replaces UDP-GlcNAc with GlcNAc. Hydrolytic cleavage of O-GlcNAc from the modified protein is catalyzed by a glycoside hydrolase designated as O-GlcNAcase or OGA.

[0004] In brain slices of mice during culture or with activated metabolism, the phosphorylation and O-GlcNAcylation of tau change in an inverse manner. From this interaction, it can be inferred that the phosphorylation and O-GlcNAcylation of tau are in a dynamic equilibrium. The presence of phosphorylated and O-GlcNAcylated serine or threonine residues allows such residues to exist in one of three different states (phosphorylated, glycosylated, free hydroxylated). The formation of such states is regulated by appropriate enzymes. Also, soluble tau in the brains of diseased individuals has even less O-GlcNAc, and insoluble tau aggregates are seen to be completely lacking in O-GlcNAc. Since O-GlcNAc is sensitive to glucose availability, such decreases and deficiencies were presumed to be due to brain glucose metabolism disorders found in AD patients. The hyperphosphorylation of tau can be traced to a decrease in O-GlcNAc due to a decreased influx of the hexosamine biosynthetic pathway (HBSP) and the resulting soluble decrease in UDP-GlcNAc. The tau O-GlcNAc level may decrease and tau may be hyperphosphorylated due to decreased OGT activity or unregulated O-GlcNAcase. The gene encoding O-GlcNAcase is present in a pseudogene on chromosome 10q24.1, which is associated with an increased risk of late-onset AD, and this is consistent with the last hypothesis mentioned above. Regardless of the origin of the decreased O-GlcNAc in the AD brain, increasing the tau O-GlcNAc level can block the hyperphosphorylation of tau and generally open up the possibility of preventing the accumulation of toxic tau species.

[0005] The attenuation of tau phosphorylation levels is thought to provide a pathway to actually slow or interrupt the progression of the disease in AD patients, and thus much effort is currently focused on the development of kinase inhibitors for therapeutic effects. Considering that lower O-GlcNAc levels appear in tau in the brains of AD patients than in normal brains, it can be considered an alternative approach to restrict tau phosphorylation by utilizing the dynamic equilibrium between O-GlcNAcylation and phosphorylation. By suppressing O-GlcNAcase in vivo, the O-GlcNAc level must increase, while the tau phosphorylation level must decrease.

[0006] Several highly potent O-GlcNAcase inhibitors have been discovered, but they have various limitations to eukaryotic O-GlcNAcase compared to functionally related eukaryotic enzymes. Such limitations include appropriate selectivity and chemical stability. Also, the point that a considerable amount is required to obtain the necessary amount was one of the limitations. One inhibitor, GlcNAcstatin, was revealed to have picomolar efficacy against the bacterial homolog of O-GlcNAcase from Clostridium perfringens, but has not yet been tested against any eukaryotic O-GlcNAcase. However, a structurally related inhibitor, gluco-nagstatin, was revealed to be active against human O-GlcNAcase with a Ki of 420 nM. The reciprocal nature of O-GlcNAc and tau phosphorylation, and the possibility of utilizing this relationship in vivo to restrict tau phosphorylation, is an interesting topic as it may trigger strategies to restrict tau hyperphosphorylation in AD.

[0007] In vitro studies using the inhibitor O-(2-acetamido-2-deoxy-D-glycopyranosylidene)amino-N-phenylcarbamate (PUGNAc) have suggested that this might be possible, but it is not selective and cannot cross the blood-brain barrier. Therefore, there was a need for an inhibitor that was more potent than existing compounds while easily crossing the blood-brain barrier and was highly stable and selective. Initial studies of human O-GlcNAcase have revealed that this enzyme uses a catalytic mechanism involving the transient formation of a substrate-assisted catalytic and non-covalently bound oxazoline intermediate from the 2-acetamido group. NAG-thiazoline, which is superficially similar to this intermediate, is a potent inhibitor of O-GlcNAcase because it is geometrically similar in the transition state. By varying most of the thiazoline substituents, a potent inhibitor of O-GlcNAcase (Ki 1 / 4 600 nM) that is 800-fold more selective for human O-GlcNAcase than human lysosomal β-hexosaminidase at pH 7.4 is produced. These thiazolines have excellent selectivity and reasonable efficacy and have limited chemical stability in solution over extended periods of days to weeks.

Prior Art Documents

Patent Documents

[0008]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0009] In order to solve the above problems, the present inventors have confirmed the O-GlcNAcase inhibitory activity of novel compounds and found that they inhibit the phosphorylation of tau, thus completing the present invention.

[0010] Therefore, an object of the present invention is to provide a novel compound having O-GlcNAcase inhibitory activity, a pharmaceutical composition thereof, and uses as a functional health food.

Means for Solving the Problems

[0011] In order to achieve the above object, the present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

[0012]

Chemical Formula

[0013] The present invention also provides a pharmaceutical composition for treating or preventing a disease caused by hyperphosphorylation of tau, which contains the compound according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0014] The present invention also provides a functional health food for improving or preventing a disease caused by hyperphosphorylation of tau, which contains the compound according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

Effects of the Invention

[0015] The novel compound of the present invention has a potent eukaryotic O-GlcNAcase inhibitory efficacy and can effectively reduce the phosphorylation of tau in vivo, thus providing various uses as a therapeutic agent for tau-related diseases.

Brief Description of the Drawings

[0016]

Figure 1

Modes for Carrying Out the Invention

[0017] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description, for technologies well-known and famous to those skilled in the art, detailed descriptions thereof can be omitted. Also, when explaining the present invention, if it is determined that specific descriptions of related known functions or configurations may shift the gist of the present invention unnecessarily, detailed descriptions thereof can be omitted. Further, the terminology used in this specification is the terminology used to appropriately express the preferred embodiments of the present invention, and this can be changed depending on the intention of the user, operator, or the convention in the field to which the present invention belongs.

[0018] Therefore, the meaning of this term must be determined based on the content throughout this specification. Throughout the specification, when a certain part states that a certain component "includes", this means that, unless otherwise stated to the contrary, it does not exclude other components and may further include other components.

[0019] Throughout the specification of the present application, the meaning of the term "aromatic ring" means including at least one aromatic ring, and the meaning of "aromatic heterocycle" means including at least one aromatic ring and at least one heterocycle.

[0020] Throughout this specification, the term "hetero element" means an element other than carbon and hydrogen elements, and means, for example, an element selected from the group consisting of N, O, S, and P, but is not limited thereto.

[0021] The active substance of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. The expression "pharmaceutically acceptable salt" means an organic or inorganic addition salt of the basic compound of the active substance, which has a relatively non-toxic and harmless effective action on the patient at a concentration, and the side effects caused by this salt do not reduce the efficacy of the basic compound of the active substance. These salts may use inorganic acids and organic acids as the free acid. As the inorganic acids, hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc. may be used. As the organic acids, citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid, etc. may be used. Further, these salts include alkali metal salts (such as sodium salt, potassium salt, etc.) and alkaline earth metal salts (such as calcium salt, magnesium salt, etc.).For example, acid addition salts include acetate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, maleate, maleate, malonate, mesylate, methyl sulfate, naphthylate, 2-naphthylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, saccharate, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, zinc salts, etc. may be included, and hydrochloride or trifluoroacetate among these is preferred.

[0022] The acid addition salts according to the present invention can be produced by a usual method, for example, dissolving the active substance in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., adding an organic acid or an inorganic acid, filtering and drying the produced precipitate, or drying after distilling off the solvent and the excess acid under reduced pressure, or crystallizing under an organic solvent.

[0023] Also, pharmaceutically acceptable metal salts can be made using a base. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to produce sodium, potassium or calcium salts as the metal salts. Also, the corresponding silver salts are obtained by reacting the alkali metal or alkaline earth metal salts with a suitable silver salt (for example, silver nitrate).

[0024] Furthermore, the present invention includes not only the active substance and its pharmaceutically acceptable salts, but also all possible solvates, hydrates, isomers, optical isomers, etc. that can be produced therefrom.

[0025] The present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof:

[0026]

Chemical formula

[0027] In one embodiment of the present invention, the compound of formula (I) may be produced by chemically covalently bonding lipoic acid and naringenin via a linker. Specifically, various novel compounds with different bonding positions can be produced by selectively reacting one of the hydroxyl groups of naringenin that reacts with lipoic acid, and in particular, various compound structures can be produced by a multi-step reaction using many linkers such as ethers and amides.

[0028] In one embodiment of the present invention, the compound of formula (I) may include compounds represented by the following formula (II) to formula (IV):

[0029] [Chemistry]

[0030] [Chemistry]

[0031] [Chemistry] In the above chemical formula, A, B and C are each independently an optionally substituted C3 to C10 cycloalkyl, an optionally substituted 5-membered unsaturated or aromatic ring, an optionally substituted 6-membered unsaturated or aromatic ring, an optionally substituted 5-membered unsaturated or aromatic heterocycle, and an optionally substituted 6-membered unsaturated or aromatic heterocycle, or a polycycle in which two or more rings selected from the above group are fused. R1, R2, R3 and n are the same as defined in the above formula (I).

[0032] In one embodiment of the present invention, in the formula (II), A may be selected from, but not limited to, the following substituents:

[0033] [Chemistry]

[0034] In one embodiment of the present invention, the compound of the formula (II) may include, but not limited to, the compounds in Tables 1 to 4 below:

[0035] [Table 1]

[0036] [Table 2]

[0037]

Table 3

[0038]

Table 4

[0039] Preferably, the A-34 compound may be a compound of the (S)-form among the enantiomers, and may be the compound A-34S represented by the following formula (V).

[0040]

Chemical formula

[0041] In one embodiment of the present invention, in the formula (III), B may be selected from the following substituents, but is not limited thereto.

[0042]

Chemical formula

[0043] In one embodiment of the present invention, the compound of the formula (III) may include, but is not limited to, the compounds in Tables 5 to 6 below:

[0044]

Table 5

[0045]

Table 6

[0046] In one embodiment of the present invention, in the formula (IV), C may be selected from the following substituents, but is not limited thereto.

[0047] [Chemical]

[0048] In one embodiment of the present invention, the compound of formula (IV) may include, but is not limited to, the compounds in Table 7 below:

[0049] [Table 7]

[0050] In one embodiment of the present invention, the compound may have, but is not limited to, O-GlcNAcase inhibitory activity.

[0051] The present invention also provides a pharmaceutical composition for treating or preventing diseases caused by hyperphosphorylation of tau, which contains the compound according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0052] In one embodiment of the present invention, the diseases may include, but are not limited to, stroke, apoplexy, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeldt-Jakob disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), progressive supranuclear palsy (PSAP), corticobasal degeneration (CBD), Lewy body disease, etc.

[0053] In the pharmaceutical composition of the present invention, the compound according to the present invention may be administered in a suitable dosage form together with carriers and diluents known in the art, and may be administered orally or parenterally by the intended method, for example, in dosage forms such as intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, suppository, etc.

[0054] The dosage form can be produced by a conventional method using suitable excipients, fillers, binders, wetting agents, disintegrants, lubricants, surfactants, dispersants, buffers, preservatives, solubilizing agents, disinfectants, sweeteners, spices, analgesics, stabilizers, isotonic solutions, etc. commonly used in pharmaceutical compositions.

[0055] Each of the above-described dosage forms may contain a pharmaceutically acceptable carrier or additive. Specific examples of the carrier or additive include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidine, carboxyvinyl polymer, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerol, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, and lactic acid. One or more additives may be selected or appropriately combined depending on the dosage form. Furthermore, as a method of administering the cell therapy agent, in addition to ordinary systemic administration such as intravenous and intraarterial administration, local administration to target cells may be performed, and an administration method combined with catheter technology and surgical operation may be used.

[0056] The composition of the present invention may contain the compound according to the present invention in a pharmaceutically effective amount together with a pharmaceutically acceptable carrier.

[0057] In the present invention, the "pharmaceutically effective amount" refers to the amount of the active ingredient that exhibits a mitigating, inhibitory, ameliorating, and / or curative effect on the immune rejection disease to be treated. The dosage of the compound according to the present invention varies depending on factors such as the patient's body weight, age, gender, health status, diet, administration time, administration method, and severity of the disease. For example, the therapeutically effective dosage can initially be determined using in vitro analysis through cell culture. In the art, the therapeutically effective amount can be determined without excessive experimentation, and such information can be used to more accurately determine the dosage useful for humans. For example, the compound according to the present invention may be administered in an amount of 0.1 to 100 mg / kg / day as the active ingredient.

[0058] The present invention also provides a functional health food for improving or preventing diseases caused by hyperphosphorylation of tau containing the compound according to the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0059] The "food" means a natural product or processed product containing one or more nutrients, preferably a product that has undergone a certain processing step and can be eaten directly. In the ordinary sense, it includes all foods, food additives, functional foods, and beverages.

[0060] Examples of foods to which the food composition can be added include various foods, beverages, gums, teas, vitamin complexes, functional foods, etc. Furthermore, special nutritional foods (e.g., prepared milks, infant foods, etc.), processed meat products, fish products, tofu products, konjac products, noodles (e.g., ramen, noodles, etc.), breads, health supplements, seasoned foods (e.g., soy sauce, miso, kochujang, mixed sauces, etc.), sauces, confectioneries (e.g., snacks), candies, chocolates, gums, ice creams, milk processed products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchis, pickles, etc.), beverages (e.g., fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.), but are not limited thereto. The foods, beverages, or food additives can be manufactured by ordinary manufacturing methods.

[0061] In addition, the "functional food" or "health functional food" means a group of foods or food compositions to which value is added by using physical, biochemical, biotechnological methods, etc. to cause the functions of the corresponding foods to act and be expressed for specific purposes, and which are designed and processed so that the in-vivo regulatory functions related to biological defense rhythm regulation, disease prevention and recovery, etc. are fully expressed in the living body. Specifically, it may be a health functional food. The functional food may contain food auxiliary additives acceptable in food science, and may further contain appropriate carriers, excipients and diluents usually used in the production of functional foods.

[0062] The types of the health supplements may be, but are not limited to, powder, granule, tablet, capsule or beverage form.

[0063] The present invention also provides a method for preventing or treating a disease caused by hyperphosphorylation of tau, which includes administering to an individual a compound according to the present invention or a pharmaceutical salt thereof, or a pharmaceutical composition containing the same.

[0064] The individual may be a mammal, for example, but not limited to, a human.

[0065] Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are for more specifically explaining the present invention, and the scope of the present invention is not limited to these examples.

Examples

[0066] Production of the compound of formula (II) The compound of the following formula (II) was produced by the following steps.

[0067]

Chemical formula

[0068] Synthesis of Schemes 1.A-1 to A-41

[0069]

Chem.

[0070]

Chem.

[0071] Reagents and conditions: (i) paraformaldehyde, AcOH, 100 °C, 4 h; (ii) TFA, DCM, r.t., overnight; (iii) TEA, DCM, 0 °C - r.t., 4 h; (iv) NaH, iodomethane, DMF, 0 °C - r.t., 1 h; (v) Amine compounds, K2CO3, acetonitrile, r.t., overnight.

[0072] Synthesis of Scheme 2.1c

[0073]

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[0074]

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[0075]

Chemical formula

[0076]

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[0077]

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[0078]

Chemical Structure

[0079]

Chemical Structure

[0080]

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[0081]

Chem.

[0082]

Chem.

[0083]

Chem.

[0084]

Chem.

[0085]

Chem.

[0086]

Chem.

[0087]

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[0088]

Chem.

[0089]

Chem.

[0090]

Chem.

[0091]

Chem.

[0092]

Chemical Structure

[0093]

Chemical Structure

[0094] [Chemical formula] 2-(4-((2-acetamidothiazol-5-yl)methyl)piperazin-1-yl)-N-(4-methoxyphenethyl)acetamide (A-8): After carrying out the synthesis method of compound A-1 using the intermediate 5a8 (200 mg, 0.88 mmol), the intermediate 3b (375 mg, 1.06 mmol), and K2CO3 (166 mg, 1.06 mmol) in acetonitrile, it was purified by column chromatography (DCM:MOH = 30:1) to obtain the compound A-8 as a pale yellow solid (117 mg, 31%). Mp: 177 - 179 °C. 1HNMR (400 MHz, CDCl3) δ: 11.21 (s, 1H), 7.20 (d, J = 1.0 Hz, 1H), 7.13 - 7.09 (m, 2H), 6.85 - 6.83 (m, 2H), 3.79 (s, 3H), 3.64 (d, J = 1.0 Hz, 2H), 3.52 (q, J = 6.6 Hz, 2H), 2.95 (s, 2H), 2.77 (t, J = 6.8 Hz, 2H), 2.44 (d, J = 4.4 Hz, 4H), 2.39 (s, 4H), 2.30 (s, 3H).

[0095]

Chem.

[0096]

Chem.

[0097] [Chemical formula] 2-chloro-N-(3,4-dimethoxyphenyl)acetamide (5a10): The synthesis method of 5a1 was carried out using 3,4-dimethoxyaniline (500 mg, 3.26 mmol), triethylamine (363 mg, 3.59 mmol), and chloroacetyl chloride (405 mg, 3.59 mmol) in DCM, and then purified by column chromatography (n-hexane / EtOAc = 1:1) to obtain intermediate 5a10 as a white solid (742 mg, 99%). 11H NMR (400 MHz, CDCl3) δ: 8.14 (s, 1H), 7.28 (d, J = 2.5 Hz, 1H), 6.96 (dd, J = 8.6, 2.5 Hz, 1H), 6.84 (d, J = 8.6 Hz, 1H), 4.19 (s, 2H), 3.90 (s, 3H), 3.88 (s, 3H).

[0098]

Chem.

[0099]

Chem.

[0100]

Chemical Structure

[0101]

Chemical Structure

[0102]

Chemical formula

[0103]

Chemical formula

[0104] [Chemical formula] 2-(4-((2-Acetamidothiazol-5-yl)methyl)piperazin-1-yl)-N-(2-bromo-4-chlorophenyl)acetamide (A-13): After implementing the synthesis method of compound A-1 using intermediate 5a13 (200 mg, 0.71 mmol), intermediate 3b (298 mg, 0.85 mmol), and K2CO3 (117 mg, 0.85 mmol) in acetonitrile, it was purified by column chromatography (DCM:MOH = 40:1) to obtain the pale yellow solid compound A-13 (110 mg, 32%). Mp: 212 - 213 °C. 1HNMR (400 MHz, CDCl3) δ: 11.25 (s, 1H), 9.94 (s, 1H), 8.44 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 2.3 Hz, 1H), 7.29 (dd, J = 8.9, 2.4 Hz, 1H), 7.22 (s, 1H), 3.72 (d, J = 0.9 Hz, 2H), 3.18 (s, 2H), 2.69 (s, 4H), 2.63 (s, 4H), 2.31 (s, 3H).

[0105]

Chem.

[0106]

Chem.

[0107] [Chemical formula] 2-chloro-N-(pyridin-2-yl)acetamide (5a15): After implementing the synthesis method of 5a1 using 2-aminopyridine (500 mg, 5.31 mmol), triethylamine (591 mg, 5.84 mmol), and chloroacetyl chloride (660 mg, 5.84 mmol) in DCM, it was purified by column chromatography (n-hexane / EtOAc = 1:1) to obtain intermediate 5a15 as a white solid (475 mg, 52%). 1HNMR (400 MHz, CDCl3) δ: 8.84 (s, 1H), 8.32 (ddd, J = 4.9, 1.9, 1.0 Hz, 1H), 8.20 (d, J = 8.4 Hz, 1H), 7.74 (ddd, J = 8.3, 7.3, 1.9 Hz, 1H), 7.11 (ddd, J = 7.4, 4.9, 1.0 Hz, 1H), 4.20 (s, 2H).

[0108]

Chem.

[0109]

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[0110]

Chemical Structure

[0111]

Chem.

[0112]

Chem.

[0113] [Chemical formula] 2-chloro-N-(4-methylpyridin-2-yl)acetamide (5a18): The synthetic method of 5a1 was carried out using 2-amino-4-methylpyridine (500 mg, 4.62 mmol), triethylamine (514 mg, 5.09 mmol), and chloroacetyl chloride (574 mg, 5.13 mmol) in DCM, and then purified by column chromatography (n-hexane / EtOAc = 1:1) to obtain intermediate 5a18 as a white solid (725 mg, 85%). 1HNMR (400 MHz, CDCl3) δ: 8.86 (s, 1H), 8.17 (dd, J = 5.1, 0.8 Hz, 1H), 8.04 (s, 1H), 6.93 (ddd, J = 5.2, 1.6, 0.8 Hz, 1H), 4.19 (s, 2H), 2.39 (d, J = 0.7 Hz, 3H).

[0114]

Chem.

[0115]

Chem.

[0116]

Chemical formula

[0117]

Chem.

[0118]

Chem.

[0119]

Chem.

[0120]

Chem.

[0121]

Chemical Structure

[0122]

Chem.

[0123]

Chem.

[0124]

Chemical formula

[0125]

Chem.

[0126]

Chem.

[0127]

Chemical Structure

[0128]

Chem.

[0129]

Chem.

[0130]

Chemical formula

[0131]

Chem.

[0132]

Chem.

[0133] [Chemical formula] 2-chloro-N-(6-methoxybenzo[d]thiazol-2-yl)acetamide (5a28): Using 2-amino-6-methoxybenzothiazole (500 mg, 2.77 mmol), triethylamine (308 mg, 3.05 mmol), and chloroacetyl chloride (344 mg, 3.05 mmol) in DCM, the synthesis method of 5a1 was carried out, and then purified by column chromatography (n-hexane / EtOAc = 5:1) to obtain white solid intermediate 5a28 (425 mg, 60%). 1HNMR (400 MHz, CDCl3) δ: 9.66 (s, 1H), 7.71 (d, J = 8.9 Hz, 1H), 7.30 (d, J = 2.6 Hz, 1H), 7.07 (dd, J = 8.9, 2.6 Hz, 1H), 4.30 (s, 2H), 3.88 (s, 3H).

[0134]

Chem.

[0135]

Chem.

[0136]

Chemical formula

[0137]

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[0138]

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[0139]

Chemical formula

[0140]

Chemical formula

[0141] [Chemical formula] 2-(4-((2-acetamidothiazol-5-yl)methyl)-3-methylpiperazin-1-yl)-N-phenylacetamide (A-32): After carrying out the synthesis method of compound A-1 using intermediate 5a1 (255 mg, 1.51 mmol), intermediate 3c (460 mg, 1.81 mmol), and K2CO3 (250 mg, 1.81 mmol) in acetonitrile, it was purified by column chromatography (DCM:MOH = 30:1) to obtain compound A-32 as a white solid (173 mg, 30%). Mp: 174 - 176 °C. 1HNMR (400 MHz, CDCl3) δ: 10.91 (s, 1H), 9.06 (s, 1H), 7.56 - 7.54 (m, 2H), 7.35 - 7.31 (m, 2H), 7.22 (s, 1H), 7.13 - 7.08 (m, 1H), 4.04 (d, J = 14.3 Hz, 1H), 3.70 (d, J = 14.5 Hz, 1H), 3.10 (d, J = 2.8 Hz, 2H), 2.84 - 2.81 (m, 1H), 2.75 - 2.72 (m, 2H), 2.62 (s, 1H), 2.54 - 2.40 (m, 2H), 2.31 (s, 3H), 2.26 - 2.25 (m, 1H), 1.19 (d, J = 6.2 Hz, 3H).

[0142]

Chem.

[0143] [Chemical formula] 2-(4-((2-acetamidothiazol-5-yl)methyl)-3-methylpiperazin-1-yl)-N-(4-chlorophenyl)acetamide (A-34): After carrying out the synthesis method of compound A-1 using intermediate 5a12 (200 mg, 0.98 mmol), intermediate 3c (295 mg, 1.16 mmol), and K2CO3 (163 mg, 1.16 mmol) in acetonitrile, it was purified by column chromatography (DCM:MOH = 50:1) to obtain white solid compound A-34 (120 mg, 30%). Mp: 184 - 187 °C. 1HNMR (400 MHz, CDCl3) δ: 12.51 (d, J = 11.3 Hz, 1H), 9.09 (s, 1H), 7.54 - 7.48 (m, 2H), 7.30 - 7.26 (m, 2H), 7.23 (s, 1H), 4.05 (d, J = 14.5 Hz, 1H), 3.72 (d, J = 14.6 Hz, 1H), 3.11 (d, J = 2.9 Hz, 2H), 2.84 (dt, J = 11.6, 2.9 Hz, 1H), 2.73 (dt, J = 10.2, 4.3 Hz, 2H), 2.66 - 2.60 (m, 1H), 2.54 - 2.41 (m, 2H), 2.34 (s, 3H), 2.30 - 2.24 (m, 1H), 1.19 (d, J = 6.1 Hz, 3H).

[0144]

Chem.

[0145] Scheme 3. Synthesis of Intermediate 4a31

[0146]

Chem.

[0147]

Chem.

[0148]

Chemical Structure

[0149]

Chemical Structure

[0150]

Chem.

[0151]

Chem.

[0152]

Chemical formula

[0153]

Chem.

[0154]

Chem.

[0155]

Chemical Structure

[0156] Scheme 4. Synthesis of Intermediate 4a33

[0157] [Chemical Structure] Reagents and conditions: (i) LiAlH4, THF, 0 °C - r.t., 1.5 h.

[0158] [Chemical Structure] [1,1’-biphenyl]-4-ylmethanamine (4a33): Lithium aluminium hydride (424 mg, 11.16 mmol) was added portionwise to a solution of 4-cyanobiphenyl (500 mg, 2.79 mmol) in THF (30 mL) at 0 °C. The reaction mixture was stirred at room temperature for 0.5 h. Extraction with DCM gave the intermediate 4a33 as a white solid (450 mg, 88%). 1HNMR (400 MHz, CDCl3) δ: 7.58 (ddt, J = 7.6, 6.3, 1.7 Hz, 4H), 7.45 - 7.32 (m, 5H), 3.92 (s, 2H).

[0159]

Chem.

[0160]

Chem.

[0161]

Chemical Structure

Example

[0162] Synthesis of the compound of formula (III) The compound of the following formula (III) was produced by the following steps.

[0163]

Chem.

[0164] Synthesis of Schemes 5.B-1 to B-12

[0165]

Chem.

[0166]

Chem.

[0167] [Chemistry] 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (b1): To a suspension of 4-piperidine carboxylic acid (500 mg, 3.87 mmol) and K2CO3 (1.07 g, 7.74 mmol) in water (8 mL) at 0 °C was added dropwise di-tert-butyl dicarbonate (845 mg, 3.87 mmol) in THF (8 mL). The reaction mixture was stirred overnight at room temperature. THF was removed in vacuo, and the aqueous layer was washed with DCM and then acidified to pH = 4 with 1N HCl. Filtration gave the white solid intermediate b1 (770 mg, 87%). 1 HNMR (400 MHz, CDCl3): δ: 4.02 (d, J = 13.1 Hz, 2H), 2.86 (t, J = 12.5 Hz, 2H), 2.49 (tt, J = 10.9, 3.9 Hz, 1H), 1.65 (dtd, J = 13.4, 11.2, 4.2 Hz, 2H), 1.46 (s, 9H).

[0168] [Chemistry] tert-butyl 4-(phenylcarbamoyl)piperidine-1-carboxylate (b2a): To a solution of aniline (122 mg, 1.31 mmol) and DMAP (208 mg, 1.70 mmol) in DCM (30 mL) at room temperature was added in one portion intermediate b1 and in one portion EDCI (326 mg, 1.70 mmol). The reaction mixture was stirred overnight at room temperature. The reaction was quenched with TH2O and extracted with DCM. The DCM layer was washed with brine, dried over NaSO4, filtered in vacuo, and concentrated. Purification by flash column chromatography (DCM:MeOH = 98:2) gave the white solid intermediate b2a (374 mg, 94%). 1HNMR (400 MHz, CDCl3): δ: 7.51 (d, J = 7.9 Hz, 2H), 7.32 (dd, J = 8.5, 7.4 Hz, 2H), 7.18 (s, 1H), 7.11 (t, J = 7.4 Hz, 1H), 4.19 (bs, 2H), 2.79 (t, J = 12.6 Hz, 2H), 2.38 (tt, J = 11.5, 3.8 Hz, 1H), 1.75 (dtd, J = 13.3, 11.8, 4.4 Hz, 2H), 1.47 (s, 9H).

[0169]

Chem.

[0170]

Chem.

[0171] [Chemical formula] tert - butyl 4 - ((4 - chlorophenyl)carbamoyl)piperidine - 1 - carboxylate (b2b): After carrying out the synthetic method of intermediate b2a using intermediate b1 (500 mg, 2.18 mmol), 4 - chloroaniline (278 mg, 2.18 mmol), EDCI (502 mg, 2.62 mmol), and DMAP (320 mg, 2.62 mmol) in DCM, it was purified by flash column chromatography (n - hexane:EtOAc = 5:1) to afford the intermediate b2b as a white solid (645 mg, 87%). 1HNMR (400 MHz, CDCl3) δ: 7.51 - 7.43 (m, 2H), 7.30 - 7.27 (m, 2H), 7.18 (s, 1H), 4.18 (bs, 2H), 2.79 (t, J = 12.8 Hz, 2H), 2.37 (tt, J = 11.5, 3.8 Hz, 1H), 1.90 (d, J = 12.4 Hz, 2H), 1.74 (dtd, J = 13.3, 11.8, 4.4 Hz, 2H), 1.47 (s, 9H).

[0172]

Chem.

[0173]

Chem.

[0174]

Chemical formula

[0175]

Chem.

[0176]

Chem.

[0177]

Chemical formula

[0178]

Chem.

[0179]

Chem.

[0180]

Chemical formula

[0181]

Chem.

[0182]

Chem.

[0183]

Chemical formula

[0184]

Chem.

[0185]

Chem.

[0186]

Chemical formula

[0187]

Chem.

[0188]

Chem.

[0189]

Chemical Structure

[0190]

Chem.

[0191]

Chem.

[0192] Synthesis of Schemes 6.4i - 4j

[0193]

Chem.

[0194]

Chem.

[0195]

Chemical formula

[0196]

Chemical formula

[0197]

Chemical Structure

[0198]

Chem.

[0199]

Chem.

[0200]

Chemical formula

[0201]

Chem.

[0202]

Chem.

[0203]

Chem.

[0204]

Chemical Structure

[0205]

Chem.

[0206]

Chem.

[0207]

Chem.

[0208]

Chem.

[0209]

Chem.

[0210]

Chemical formula

[0211]

Chem.

[0212]

Chem.

[0213]

Chemical formula

[0214]

Chemical formula

[0215]

Chem.

[0216]

Chem.

[0217] [Chemical formula] 1 - ((2 - acetamidothiazol - 5 - yl)methyl)-N-(1-(naphthalen - 2 - yl)ethyl)piperidine - 4 - carboxamide (B - 12): Synthesis method of compound b - 1 was carried out using 2 - acetamidothiazole (73 mg, 0.51 mmol), b3l (130 mg, 0.34 mmol), and paraformaldehyde (51 mg, 1.71 mmol) in acetic acid (20 mL), and then purified by flash column chromatography (DCM:MeOH = 50:1) to obtain white solid compound B - 12 (75 mg, 50%). 11H NMR (400 MHz, CDCl3) δ: 11.28 (s, 1H), 7.85 - 7.71 (m, 4H), 7.51 - 7.38 (m, 3H), 7.20 (s, 1H), 5.79 (s, 1H), 5.35 - 5.24 (m, 1H), 3.70 (s, 2H), 3.00 (s, 2H), 2.28 (s, 3H), 1.58 (d, J = 6.9 Hz, 3H).

[0218] Scheme 7. Synthesis of SB - 13 to B - 19

[0219]

Chem.

[0220]

Chem.

[0221]

Chem.

[0222]

Chem.

[0223] [Chemical formula] 1-((2-Acetamidothiazol-5-yl)methyl)-N-(p-tolyl)piperidine-4-carboxamide (B-15): After implementing the synthesis method of compound B-14 using p-toluidine (74 mg, 0.69 mmol), b5a (200 mg, 0.69 mmol), EDCI (159 mg, 0.83 mmol), HOBt (113 mg, 0.83 mmol), and DIPEA (223 mg, 1.73 mmol) in DMF (30 mL), it was purified by flash column chromatography (DCM:MeOH = 40:1) to obtain compound B-15 as a white solid (28 mg, 11%). Mp: 262 - 264 °C. 1 HNMR (400 MHz, DMSO-d6): δ: 11.93 (s, 1H), 9.72 (s, 1H), 7.49 - 7.44 (m, 2H), 7.25 (s, 1H), 7.10 - 7.05 (m, 2H), 3.62 (s, 2H), 2.89 (dd, J = 7.7, 3.9 Hz, 2H), 2.34 - 2.26 (m, 1H), 2.23 (s, 3H), 2.12 (s, 3H), 2.02 - 1.90 (m, 2H), 1.77 - 1.58 (m, 4H).

[0224] [Chemical formula] 1-(Benzo[d][1,3]dioxol-5-yl)ethyl 1-((2-acetamidothiazol-5-yl)methyl)piperidine-4-carboxylate (B-13): After implementing the synthesis method of compound B-14 using intermediate b5a (400 mg, 1.38 mmol), 2i (230 mg, 1.38 mmol), EDCI (318 mg, 1.66 mmol), and HOBt (225 mg, 1.66 mmol) in DMF, it was purified by flash column chromatography (DCM:MeOH = 50:1) to obtain compound B-13 as a white solid (50 mg, 8%). Mp: 215 - 217 °C. 1HNMR (400 MHz, DMSO-d6): δ: 11.93 (s, 1H), 7.23 (s, 1H), 6.93 - 6.79 (m, 3H), 6.00 (d, J = 5.5 Hz, 2H), 5.72 (dt, J = 13.1, 6.7 Hz, 1H), 3.59 (s, 2H), 2.77 (dd, J = 10.2, 5.2 Hz, 2H), 2.30 (ddt, J = 10.7, 7.7, 3.8 Hz, 1H), 2.11 (s, 3H), 2.02 (dt, J = 11.5, 3.1 Hz, 2H), 1.78 (dd, J = 11.6, 7.2 Hz, 2H), 1.54 (tdd, J = 13.2, 8.7, 3.4 Hz, 2H), 1.41 (d, J = 6.5 Hz, 3H).

[0225]

Chem.

[0226]

Chem.

[0227]

Chem.

[0228]

Chem.

[0229]

Chem.

[0230]

Chem.

Example

[0231] Synthesis of the compound of formula (IV) The compound of formula (IV) was prepared in the following steps.

[0232]

Chem.

[0233] Synthesis of C - 1 to C - 8 in Scheme 8

[0234]

Chem.

[0235]

Chem.

[0236]

Chemistry

[0237]

Chemistry

[0238]

Chem.

[0239]

Chem.

[0240]

Chem.

[0241]

Chem.

[0242]

Chemical formula

[0243]

Chem.

[0244] [Chemistry] 2-(1-((2-acetamidothiazol-5-yl)methyl)piperidin-4-ylidene)-N-(3,4-dimethoxyphenyl)acetamide (C-6): After carrying out the synthesis method of compound B-14 using intermediate c-5 (393 mg, 1.31 mmol), 3,4-dimethoxylaniline (200 mg, 1.31 mmol), EDCI (300 mg, 1.57 mmol), and HOBt (213 mg, 1.57 mmol) in DMF, it was purified by flash column chromatography (DCM:MeOH = 20:1) to obtain compound C-6 as a white solid (20 mg, 4%). Mp: 214 - 219 °C. 1 HNMR (500 MHz, DMSO-d6): δ: 11.97 (s, 1H), 9.77 (s, 1H), 7.37 (d, J = 2.3 Hz, 1H), 7.25 (s, 1H), 7.07 (dd, J = 8.8, 2.3 Hz, 1H), 6.85 (d, J = 8.7 Hz, 1H), 5.79 (s, 1H), 3.70 (d, J = 3.8 Hz, 6H), 3.65 (s, 2H), 2.96 (s, 3H), 2.43 (d, J = 6.2 Hz, 2H), 2.25 (s, 2H), 2.10 (d, J = 3.8 Hz, 4H).

[0245] [Chemistry] 2-(1-((2-acetamidothiazol-5-yl)methyl)piperidin-4-ylidene)-N-(4-methoxybenzyl)acetamide (C-7): The synthetic method of compound B-14 was carried out using intermediate c-5 (439 mg, 1.46 mmol), 4-methoxylbenzylamine (200 mg, 1.46 mmol), EDCI (335 mg, 1.75 mmol), and HOBt (237 mg, 1.75 mmol) in DMF, and then purified by flash column chromatography (DCM:MeOH = 20:1) to obtain compound C-7 as a white solid (44 mg, 7%). Mp: 222 - 224 °C. 1 HNMR(500 MHz, DMSO-d6): δ: 11.97(s,1H),8.27(t,J = 6.0 Hz, 1H), 7.24 (s, 1H), 7.15 (d, J = 8.5 Hz, 2H), 6.86 (d, J = 8.5 Hz, 2H), 5.65 (s, 1H), 4.18 (d, J = 5.9 Hz, 2H), 3.71 (s, 3H), 3.63 (s, 2H), 2.92 (s, 2H), 2.42 (dt, J = 21.4, 5.8 Hz, 4H), 2.21 - 2.15 (m, 2H), 2.11 (s, 3H).

[0246]

Chemical formula

Example

[0247] Enzyme activity assay The human OGA (recombinant hOGA protein, advanced Protein Technologies corp., Korea) enzyme reaction was carried out in a reaction solution containing 25 mM Tris / HCl and 0.1 mg / ml bovine serum albumin (pH 7.5) using 2 mM 4-Methylumbelliferyl N-acetyl-β-D-glucosaminide (69585; Sigma) dissolved in DMSO as the substrate. The amount of human OGA enzyme used in the reaction was 8 ng / well. Before the start of the reaction, various amounts of the compounds of Examples 1 to 3 were added to the enzyme. After carrying out the reaction in a 384-well plate at 37 °C for 20 minutes, the substrate was added to start. The increase in fluorescence was measured using a SAFIRE (Tecan, Switzerland) fluorometer and detected at the excitation and emission wavelengths set at 360 nm and 460 nm, respectively. The detected enzyme activities are shown in Tables 8 to 14 below.

[0248]

Table 8

[0249]

Table 9

[0250]

Table 10

[0251]

Table 11

[0252]

Table 12

[0253]

Table 13

[0254]

Table 14

[0255] Furthermore, the inhibitory ability of A-34S and A-34R compounds corresponding to the enantiomers of the A-34 compound against human OGA enzyme at different concentrations is shown in Figure 1, and the IC50 concentration values are shown in Table 15 below.

[0256]

Table 15

[0257] As shown in Table 15 above, it was confirmed that among the two enantiomers of the A-34 compound, the inhibitory ability of the (S)-form compound of A-34 against human OGA enzyme activity was significantly excellent.

[0258] Compound name of A-34S: (S)-2-(4-((2-acetamidothiazol-5-yl)methyl)-3-methylpiperazin-1-yl)-N-(4-chlorophenyl)acetamide

[0259] Compound name of A-34R: (R)-2-(4-((2-acetamidothiazol-5-yl)methyl)-3-methylpiperazin-1-yl)-N-(4-chlorophenyl)acetamide

[0260] So far, the present invention has been studied mainly with respect to its preferred embodiments. Those having ordinary knowledge in the technical field to which the present invention pertains should be able to understand that the present invention can be modified within a range not deviating from its essential characteristics. Therefore, the disclosed embodiments should be considered from an explanatory perspective rather than a limiting perspective. Thus, the scope of the present invention is shown in the claims including the above description, and differences within an equivalent range thereto should be construed as being included in the present invention.

Industrial Applicability

[0261] The present invention relates to a novel compound having O-GlcNAcase inhibitory activity and may be useful as a pharmaceutical composition for treating diseases caused by hyperphosphorylation of tau.

Claims

1. A compound represented by the following formula (II) to (IV) or a pharmaceutically acceptable salt thereof, wherein: 【Chemical Formula 1】 【Chemical Formula 2】 【Chemical Formula 3】 In the formula (II), R1 and R2 are each independently hydrogen or C1-C5 alkyl, n is an integer from 0 to 3, X is N, A is selected from the following substituents: 【Chemical Formula 4】 In the formula (III), R3 is hydrogen or C1-C5 alkyl, n is an integer from 0 to 3, X is NH or O, B is selected from the following substituents: 【Chemical Formula 5】 In the formula (IV), n is an integer from 0 to 3, C is selected from the following substituents, a compound or a pharmaceutically acceptable salt thereof. 【Chemical Formula 6】

2. The compound of formula (II) according to claim 1, which comprises the following compounds A-1 and A-3 to A-41, or a pharmaceutically acceptable salt thereof. 【Table 1】 【Table 2】 【Table 3】 【Table 4】 【Table 5】

3. The compound of formula (III) is the compound according to claim 1 or a pharmaceutically acceptable salt thereof, which includes the following compounds B-1 to B-19. [Table 6] [Table 7]

4. The compound of formula (IV) is the compound according to claim 1 or a pharmaceutically acceptable salt thereof, which includes the following compounds C-1 to C-8. [Table 8]

5. The compound is the compound according to claim 1 or a pharmaceutically acceptable salt thereof, which has O-GlcNAcase inhibitory activity.

6. The A-34 compound is the compound A-34S represented by the following formula (V) as the (S)-form compound among enantiomers, which is characterized by the compound according to claim 2 or a pharmaceutically acceptable salt thereof. [Chemical Formula 7]

7. A pharmaceutical composition for the treatment or prevention of a disease caused by hyperphosphorylation of tau, which contains the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof as an active ingredient.

8. The disease is selected from the group consisting of stroke, apoplexy, memory loss, memory impairment, dementia, forgetfulness, Parkinson's disease, Alzheimer's disease, Pick's disease, Creutzfeldt-Jakob disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), progressive supranuclear palsy (PSAP), corticobasal degeneration (CBD), and Lou Gehrig's disease. The pharmaceutical composition for the treatment or prevention of a disease caused by hyperphosphorylation of tau according to claim 7.

9. A functional health food for improving or preventing a disease caused by hyperphosphorylation of tau, comprising, as an active ingredient, the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.

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