KR102104804B1 - Novel acetylcholinesterase inhibitory derivatives, preparation method thereof, and pharmaceutical composition for use in preventing or treating nervous system diseases containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel acetylcholinesterase inhibitor derivative, a method for preparing the same, and a pharmaceutical composition for preventing or treating a nervous system disease containing the same as an active ingredient, a compound according to the present invention, a stereoisomer thereof, or a pharmaceutical thereof As an acceptable salt, it can excellently inhibit the activity of acetylcholinesterase, such as acetylcholinesterase related diseases, such as diseases that have been correlated with acetylcholinesterase to date, such as It has a useful effect as a pharmaceutical composition for preventing or treating a nervous system disease or a health functional food composition for preventing or improving a nervous system disease.

Description

Novel acetylcholinesterase inhibitory derivatives, preparation method thereof, and pharmaceutical composition for use in preventing or treating nervous system diseases containing the same as an active ingredient}

The present invention relates to a novel acetylcholinesterase inhibitor derivative, a preparation method thereof, and a pharmaceutical composition for the prevention or treatment of a nervous system disease containing the same as an active ingredient.

Acetylcholine degrading enzyme inhibitors such as donepezil are used as a label drug for Alzheimer's dementia or Alzheimer's disease. 'Huperzine A' or its derivative 'BIS-001' is an acetylcholine inhibitor that is desirable for non-clinical studies on refractory epilepsy, and has a BBB (Blood-Brain-Barrier) transmittance and a different action from existing therapeutics It is predicting entry into the Phase 1 clinical trial, showing the mode of action (MOA).

In addition, some acetylcholine inhibitors exhibit a neuroprotective effect that expects a prophylactic effect as well as a therapeutic effect. Although acetylcholine inhibitors have been proven to prevent and treat neurological diseases, there are currently no underlying therapeutic agents (DMDs, disease modifying drugs) that will control Alzheimer's disease, and are clinically proven drugs such as donepezil and galatamine. , Tacrine, etc. have the effect of alleviating symptoms that slow the prognosis of the disease, but cannot eliminate or resolve the etiology.

In other words, the current Alzheimer's market has high unmet medical needs, as existing commercial therapies do not completely block Alzheimer's progress and cannot be used for early detection and prevention.

In recent academic papers, acetylcholine degrading enzyme inhibitors not only contribute to the increase in the initial choline concentration when used, but also increase the acetylcholine receptors in the nervous system (up-regulation) and enhance the PI3K-AKT signal transduction system to suppress cell death, etc. It has been reported that it contributes to the treatment of glutamate neurotoxic effects by the mechanism of (non-patent document 1).

Thus, despite the limitations of current clinical drugs, the drug class still has the value of developing new drugs. In particular, since the drug group of the same mechanism of action (MOA) has a parental nucleus different from the existing therapeutic agent, the development of an acetylcholine inhibitor of the new parental nucleus is further required because it has a unique pharmacological profiling.

While trying to develop an acetylcholine degrading enzyme inhibitor having a differential parental structure, the inventor built a 'in silico pharmacological model' on a computerized computer, predicted activity through a virtual search, and attempted a method of selecting a compound. , By selecting a novel acetylcholinesterase inhibitor derivative according to the present invention, and confirming that acetylcholinesterase can be excellently inhibited, preventing acetylcholinesterase related diseases, for example, nervous system disease or The present invention was completed by confirming that it can be used as an active ingredient of a therapeutic pharmaceutical composition.

Akaike, Akinori, et al. Journal of Molecular Neuroscience 40.1-2 (2010): 211-216.

It is an object of the present invention to provide novel inhibitory compounds for acetylcholinesterase, stereoisomers thereof and pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide a method for designing the acetylcholinesterase inhibitory compound.

Another object of the present invention is to provide a method for predicting activity of the acetylcholinesterase inhibitory compound.

Another object of the present invention is to provide a method for preparing the acetylcholinesterase inhibitory compound, a stereoisomer thereof, and a pharmaceutically acceptable salt thereof.

Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of acetylcholinesterase-related diseases containing the acetylcholinesterase inhibitory compound, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient Is to do.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of neurological diseases containing the acetylcholinesterase inhibitory compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is an acetylcholinesterase inhibitory compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, as an active ingredient, an acetylcholinesterase-related disease preventing or improving health functional food composition Is to provide.

Another object of the present invention is the acetylcholinesterase inhibitory compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, comprising administering to a subject in a therapeutically effective amount (subject). It is to provide a method of treatment.

In order to achieve the above object,

The present invention provides a compound represented by Formula 1 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

R ¹ is the group consisting of -H, substituted or unsubstituted C _1-10 ring of a linear or branched alkyl, substituted or unsubstituted C _{6- 10} aryl -C _{1- 3} alkyl, or N, O, and S being a 5- to 10-ring heteroaryl each -C _{1- 3} alkyl-substituted or unsubstituted ring containing a hetero atom at least one member selected from,

Wherein the substituted alkyl, substituted aryl-alkyl, and substituted heteroaryl-alkyl are -OH, halogen, nitro, cyano, amino, C _1-5 straight or branched chain alkyl, and C _{1- 5} is substituted with one or more substituents selected from the group consisting of straight-chain or branched alkoxy;

When the alkyl portion in the substituted alkyl, substituted aryl-alkyl, and substituted heteroaryl-alkyl has 2 or more carbon atoms, optionally a single bond may be substituted with one or more double or triple bonds;

R ² is = O or -OH; And

R ^3's is a substituted or unsubstituted C _{6- 10} aryl, or N, O, and a substituted or unsubstituted containing one or more heteroatoms selected from the group consisting of S ring of 5 to 10, each ring heteroaryl,

Here, the substituted aryl and substituted heteroaryl are -OH, halogen, nitro, cyano, substituted or unsubstituted amino, substituted or unsubstituted C _1-5 linear or branched alkyl, substituted or unsubstituted Substituted or unsubstituted containing at least one hetero atom selected from the group consisting of substituted C _1-5 linear or branched alkoxy, substituted or unsubstituted C _3-10 cycloalkyl, N, O, and S Substituted or unsubstituted 5 comprising one or more heteroatoms selected from the group consisting of substituted 5- to 10-membered heterocycloalkyl, substituted or unsubstituted C _6-10 aryl, or N, O, and S. To 10 or more rings may be substituted with one or more substituents selected from the group consisting of heteroaryl, and when two or more substituents are present, two or more substituents may or may not be connected to each other,

Here again, the substituted amino, substituted alkyl, substituted alkoxy, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, and substituted heteroaryl are -OH, halogen, nitro, cyano, amino, It is substituted with one or more substituents selected guneu roptu consisting of alkoxy of straight- or branched-chain alkyl, and C _1-5 straight or branched-chain C _1-5.

In addition, the present invention derives a pharmacological group model from acetylcholinesterase (step 1); And

A novel acetylcholinesterase inhibitory compound comprising the step of deriving a virtual active substance by performing a virtual search using the pharmacological model derived in step 1 (step 2), that is, of the compound represented by the formula (1) Provide a design method.

Furthermore, the present invention derives a pharmacological group model from acetylcholinesterase (step 1); And

A novel acetylcholinesterase inhibitory compound, that is, a method for predicting activity of a compound represented by Formula 1, comprising the step of predicting the activity of the virtual active substance using the pharmacological model derived in Step 1 (Step 2). Is to provide

In addition, the present invention, as shown in Scheme 1 below,

Preparing a compound represented by Chemical Formula 3 from a compound represented by Chemical Formula 2 (step 1);

Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in Step 1 (Step 2); And

It provides a method for producing a compound represented by the formula (1) comprising the step (step 3) of preparing a compound represented by the formula (1) from the compound represented by the formula (4) prepared in step 2.

[Scheme 1]

In Reaction Scheme 1,

R ¹ , R ² , and R ³ are as defined in Formula 1 above.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating acetylcholinesterase related diseases containing the compound, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of a nervous system disease containing the compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a health functional food composition for preventing or improving diseases related to acetylcholinesterase containing the compound, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

In addition, the present invention provides a method for treating an acetylcholinesterase related disease, comprising administering the compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject in a therapeutically effective amount.

A compound according to the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, can excellently inhibit the activity of acetylcholinesterase, acetylcholinesterase related diseases such as acetylcholinesters to date It has a useful effect as a pharmaceutical composition for the prevention or treatment of diseases that have been correlated with a terase, for example, a prevention or treatment of a nervous system disease, or a health functional food composition for the prevention or improvement of a nervous system disease.

Figure 1 shows a model of the acetylcholinesterase pharmacological group, produced with Schrodinger's "Phase" f, to derive the structure of a novel acetylcholinesterase inhibitory compound.
Figure 2 is a graph showing the acetylcholinesterase inhibitory activity of the Example compound of the present invention.

Hereinafter, the present invention will be described in detail.

The following description is provided to aid the understanding of the invention, and the present invention is not limited to the contents of the following description.

The present invention provides a compound represented by Formula 1 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Chemical Formula 1,

R ¹ is -H, substituted or unsubstituted C _1-10 straight or branched chain alkyl, substituted or unsubstituted C _6-10 aryl-C _1-3 alkyl, or N, O, and S A substituted or unsubstituted 5- to 10-membered heteroaryl-C _1-3 alkyl containing at least one hetero atom selected from

Wherein the substituted alkyl, substituted aryl-alkyl, and substituted heteroaryl-alkyl are -OH, halogen, nitro, cyano, amino, C _1-5 straight or branched chain alkyl, and C _{1- 5} is substituted with one or more substituents selected from the group consisting of straight-chain or branched alkoxy;

When the alkyl portion in the substituted alkyl, substituted aryl-alkyl, and substituted heteroaryl-alkyl has 2 or more carbon atoms, optionally a single bond may be substituted with one or more double or triple bonds;

R ² is = O or -OH; And

R ^3's is a substituted or unsubstituted C _{6- 10} aryl, or N, O, and a substituted or unsubstituted containing one or more heteroatoms selected from the group consisting of S ring of 5 to 10, each ring heteroaryl,

Here, the substituted aryl and substituted heteroaryl are -OH, halogen, nitro, cyano, substituted or unsubstituted amino, substituted or unsubstituted C _1-5 linear or branched alkyl, substituted or unsubstituted Substituted or unsubstituted containing at least one hetero atom selected from the group consisting of substituted C _1-5 linear or branched alkoxy, substituted or unsubstituted C _3-10 cycloalkyl, N, O, and S Substituted or unsubstituted 5 comprising one or more heteroatoms selected from the group consisting of substituted 5- to 10-membered heterocycloalkyl, substituted or unsubstituted C _6-10 aryl, or N, O, and S. To 10 or more rings may be substituted with one or more substituents selected from the group consisting of heteroaryl, and when two or more substituents are present, two or more substituents may or may not be connected to each other,

Here again, the substituted amino, substituted alkyl, substituted alkoxy, substituted cycloalkyl, substituted heterocycloalkyl, substituted aryl, and substituted heteroaryl are -OH, halogen, nitro, cyano, amino, It is substituted with one or more substituents selected guneu roptu consisting of alkoxy of straight- or branched-chain alkyl, and C _1-5 straight or branched-chain C _1-5.

Preferably,

Wherein R ¹ is C _{6- 10} aryl -C _{1- 2} alkyl, or N, O, and from 5 to 10 ring heteroaryl each -C _{1- 2} alkyl containing one or more heteroatoms selected from the group consisting of S to be.

Meanwhile preferably,

,

,

, 또는

이다.R ³ is

,

,

, or

to be.

Most preferably,

Preferred examples of the compound represented by Formula 1 according to the present invention include the following compounds.

(1) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (9H-carbazol-4-yl) -4-hydroxybutan-2-one;

(2) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4-hydroxy-4- (9-methyl-9H-carbazol-4-yl) butan-2-one;

(3) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (biphenyl-3-yl) -4-hydroxybutan-2-one;

(4) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (biphenyl-4-yl) -4-hydroxybutan-2-one;

(5) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (9H-carbazol-4-yl) butane-1,3-diol;

(6) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (9-methyl-9H-carbazol-4-yl) butane-1,3-diol;

(7) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (biphenyl-3-yl) butane-1,3-diol; And

(8) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (biphenyl-4-yl) butane-1,3-diol.

On the other hand, the compounds of the present invention may exist in the form of salts, particularly pharmaceutically acceptable salts. As the salt, salts commonly used in the art may be used without limitation, such as acid addition salts formed by pharmaceutically acceptable free acids. The term "pharmaceutically acceptable salt" of the present invention is a concentration having a relatively non-toxic and harmless effective action to a patient, and any side effect of the compound whose side effects caused by this salt do not degrade the beneficial efficacy of the compound represented by Formula 1 Means all organic or inorganic addition salts.

Acid addition salts are prepared by dissolving the compound in a conventional method, for example, an aqueous solution of excess acid, and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The molar amount of the compound and the acid or alcohol in water (eg, glycol monomethyl ether) can be heated and then the mixture is evaporated to dryness or the precipitated salt can be suction filtered.

At this time, organic and inorganic acids can be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, and tartaric acid can be used as the inorganic acid, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, and maleic acid can be used as the organic acid. (maleic acid), succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid (gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, etc. can be used. , Not limited to these.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal salt or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the inexpensive compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to manufacture sodium, potassium, or calcium salts, but is not limited thereto. The corresponding silver salt can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

Pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compounds of Formula 1, unless indicated otherwise. For example, pharmaceutically acceptable salts may include sodium, calcium, and potassium salts of hydroxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, and hydrogen phosphate. , Dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, etc., and preparation of salts known in the art It can be prepared through a method.

Pharmaceutically acceptable salts of the 'N-benzyl and N-phenoxyethyl substituted 4-hydroxy-4-aryl butan-2-one' compounds of the present invention include 'N-benzyl and N-phenoxyethyl substitution Any salt can be used as long as it is a salt of a compound that exhibits a therapeutic effect of a nervous system disease by inhibiting acetylcholinesterase activity equivalent to the 4-hydroxy-4-aryl butan-2-one 'derivative compound.

In addition, the compound represented by Formula 1 according to the present invention includes, without limitation, solvates such as pharmaceutically acceptable salts thereof and possible hydrates that can be prepared therefrom. Solvates of the compound represented by Formula 1 may be prepared from a compound represented by Formula 1 using methods known in the art.

In addition, the compound represented by Formula 1 according to the present invention may be prepared in a crystalline form or an amorphous form, and when prepared in a crystalline form, may be optionally hydrated or solvated. In the present invention, a compound containing various amounts of water as well as a stoichiometric hydrate of the compound represented by Chemical Formula 1 may be included. Solvates of the compound represented by Formula 1 according to the present invention include both stoichiometric solvates and non-stoichiometric solvates.

In addition, the present invention derives a pharmacological group model from acetylcholinesterase (step 1); And

A novel acetylcholinesterase inhibitory compound comprising the step of deriving a virtual active substance by performing a virtual search using the pharmacological model derived in step 1 (step 2), that is, of the compound represented by the formula (1) Provide a design method.

Hereinafter, a method for designing a novel acetylcholinesterase inhibitory compound of the present invention, that is, a compound represented by Formula 1 will be described in detail.

Step 1 of the design method of a novel acetylcholinesterase inhibitory compound according to the present invention is a step of deriving a pharmacological model from acetylcholinesterase.

At this time, the derivation of the pharmacological group model is the enzyme (protein) targeted by the present invention, the binding site of the acetylcholinesterase and the expected inhibitory compound, the compound structure of the acetylcholinesterase and its active modulator (ligand), and In particular, by analyzing the changed binding site of acetylcholinesterase when the ligand is at the binding site, a pharmacological model is derived. Here, analysis using a computer is common, and a program that conforms to this can be used. That is, for example, but not limited to, it may be used by adopting a program that is used in a commercially available or conventional pharmacological research, and in one embodiment, in the present invention, the pharmacist model using 'Phase' of Schrödinger Was derived.

On the other hand, the pharmacological group model, even at the binding site of acetylcholinesterase, according to the amino acid sequence constituting the binding site or a substituent in contact with the outside, when the ligand (activity modulator) compound is located a compound having a desirable binding force To derive, it calculates different cohesion at each position and derives a pharmacological model based on this. In the present invention, a pharmacological group model for acetylcholinesterase was derived as shown in FIG. 2 using 'Phase' from Schrödinger.

Step 2 of the design method of the novel acetylcholinesterase inhibitory compound according to the present invention is a step of deriving a virtual active substance by performing a virtual search using the pharmacological model derived in step 1 above.

In this case, the virtual search derives a virtual active substance based on the pharmacological group model derived in step 1, or, preferably, utilizes a compound of a library, that is, a predetermined database, among them, excellent binding strength suggested by the pharmacological model It is to search for compounds expected to exhibit (active to acetylcholinesterase) to derive virtually active substances. Again, but not limited to, it is possible to use a conventional computer program, that is, a program capable of predicting or calculating activity by introducing a compound library into a pharmacological model, but is not limited thereto, in one embodiment, Schrödinger's' Phase 'Can be used to derive a virtual effective substance.

In the present invention, the compound represented by Chemical Formula 1 was designed using the above-described design method of the present invention, specifically, the pharmacological model of FIG. 1.

Furthermore, the present invention derives a pharmacological group model from acetylcholinesterase (step 1); And

A novel acetylcholinesterase inhibitory compound, that is, a method for predicting activity of a compound represented by Formula 1, comprising the step of predicting the activity of the virtual active substance using the pharmacological model derived in Step 1 (Step 2). Is to provide

The method for predicting activity of the present invention is to derive a pharmacological model as in the design method of the present invention described above, and to predict activity by substituting a compound of a library or a compound of an active candidate group. Although not limited thereto, the present invention predicted the activity of a virtual active substance, that is, a compound represented by Chemical Formula 1 using Schrodinger's 'Phase', and as a result, a novel acetylcholinesterase inhibitory compound of the present invention, namely , It was predicted that the compound represented by Formula 1 can excellently inhibit acetylcholinesterase (see Experimental Example 1 and Figure 1 below).

In addition, the present invention, as shown in Scheme 1 below,

Preparing a compound represented by Chemical Formula 3 from a compound represented by Chemical Formula 2 (step 1);

Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in Step 1 (Step 2); And

It provides a method for producing a compound represented by the formula (1) comprising the step (step 3) of preparing a compound represented by the formula (1) from the compound represented by the formula (4) prepared in step 2.

[Scheme 1]

In Reaction Scheme 1,

R ¹ , R ² , and R ³ are as defined in Formula 1 above.

Hereinafter, a method for preparing a compound represented by Chemical Formula 1 of Reaction Scheme 1 according to the present invention will be described in detail step by step.

In the method for preparing a compound represented by Chemical Formula 1 according to the present invention, step 1 is a step of preparing a compound represented by Chemical Formula 3 from a compound represented by Chemical Formula 2.

At this time, it can be understood that step 1 is a reaction of introducing a -R ¹ substituent to the terminal amino group represented by Chemical Formula 2. Here, the reaction of the above step 1 is in the desired compound of the formula If the third reaction to produce the compound represented by being included in the present invention without limitation, for but And there limiting, one example of the first step is acid catalyzed conditions, R ¹ - After the imine condensation reaction between the aldehyde and the compound represented by Chemical Formula 2, it may be obtained through reduction, and as another example, a compound represented by Chemical Formula 3 may be obtained through a substitution reaction with R ¹ -bromide. Furthermore, although not limited thereto, in one specific embodiment, step 1 may be performed by reacting a compound represented by Chemical Formula 2, R1-Br, ET ₃ N in toluene.

At this time, the solvent used in Step 1 of Reaction Scheme 1 may preferably be toluene as an example, but the solvent usable in the step is dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydrofuran (THF). ), Methylene chloride, toluene, acetonitrile and mixtures thereof.

In addition, in the above step, the reaction temperature is not particularly limited, but is preferably performed at 0 to 30 ° C, and the reaction time is not particularly limited, but it is preferable to react for 0.5-20 hours.

As another specific example in which step 1 may be performed, R ¹ -aldehyde and PTSA are reacted in toluene, and then, a reduction reaction is performed with methanol to NABH ₄ to prepare a target compound represented by Chemical Formula 3. have.

At this time, the solvent used in Step 1 of Reaction Scheme 1 may preferably be toluene as an example, but the solvent usable in the step is dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydrofuran (THF). ), Methylene chloride, toluene, acetonitrile and mixtures thereof.

In addition, in the above step, the reaction temperature is not particularly limited, but is preferably performed at 0 to 30 ° C, and the reaction time is not particularly limited, but it is preferable to react for 0.5-20 hours.

In the method for preparing a compound represented by Chemical Formula 1 according to the present invention, Step 2 is a step of preparing a compound represented by Chemical Formula 4 from a compound represented by Chemical Formula 3.

At this time, step 2 may be understood as a reaction in which a substituent extended to the terminal amino group of the compound represented by Chemical Formula 3 is introduced. Here, the reaction of step 2 is included in the present invention without limitation as long as it is a reaction capable of preparing a compound represented by the formula (4), which is a target compound, but is not limited thereto. In one specific example, step 2 is 1-chloropropane- The compound represented by Chemical Formula 4 may be prepared through the substitution reaction of Chemical Formula 3 using 2-one.

At this time, the solvent used in Step 2 of Reaction Scheme 1 may be, for example, Tetrahydrofuran, but as a solvent usable in the step, dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydro Furan (THF), methylene chloride, toluene, acetonitrile and mixtures thereof can be used.

In addition, in the above step, the reaction temperature is not particularly limited, but is preferably performed at 0 to 30 ° C, and the reaction time is not particularly limited, but it is preferable to react for 0.5-20 hours.

In the method for preparing a compound represented by Formula 1 according to the present invention, step 3 is a step for preparing a compound represented by Formula 1 from a compound represented by Formula 4.

At this time, the step 3 is a step performed to introduce a selected substituent, so as to provide the desired binding force to the acetylcholinesterase binding portion to be provided in the present invention in the terminal group of the compound represented by the formula (4) The compound represented by Formula 1 of the present invention, which can be understood and prepared by performing step 3 above, prepares a new acetylcholinesterase inhibitory compound according to the purpose of the method of the present invention.

Here, the reaction of step 3 is included in the present invention without limitation as long as it is a reaction capable of preparing a compound represented by Formula 1, that is, the target compound of the present invention, that is, a compound designed according to an acetylcholinesterase binding moiety. Although not limited, for example, step 3 may be performed through an aldol reaction of an aryl-aldehyde or heteroaryl-aldehyde with a compound represented by Chemical Formula 4, and in one embodiment, LDA (lithium diisopropylamide), It can be carried out by reacting a compound represented by the formula (4), arylaldehyde or heteroarylaldehyde in THF.

In particular, the LDA used in step 4 may use other bases, such as NaOH, NaHMDS, and may preferably select and use a conventionally known base in consideration of the production yield of the target compound. In the present invention, NaHMDS, NaOH, LDA (solid), and LDA (20% in THF) were used, respectively. As a result, the highest yield was observed in LDA (20% in THF). However, the example of the base described above is only one specific example in performing the manufacturing step of the present invention, and a base of a known technique may be used without limitation, which is included in the present invention.

Specifically, the present inventors performed a reaction by adjusting different bases, solvents, and temperatures, as shown in Table 1 below, to select the more preferable conditions of step 4, and then compared the yields of the compounds prepared therefrom.

base Temperature (℃) menstruum yield(%) One NaHMDS -78 THF 11.4 2 NaOH 0 Ethanol: water (1: 2) - 3 LDA (solid) -78 THF - 4 LDA (20% of THF) -78 THF 26

As can be seen from Table 1, it was confirmed that step 4 is preferably performed as the reaction conditions represented by number 4. As described above, a person skilled in the art changes or modifies factors caused by variables in performing the reaction, such as the above-described base, reagent, temperature, solvent, catalyst, etc., and manufactures considering the production yield of the desired compound and the shape of the target compound, etc. Method can be selected and used.

Accordingly, the manufacturing method of the present invention is to be included in the present invention without limitation, as long as it is a method capable of manufacturing the same target compound as the present invention, that is, a compound represented by Formula 1 by changing or modifying it based on known technology and common knowledge. I can understand.

At this time, the solvent used in Step 3 of Reaction Scheme 1 may be, for example, tetrahydrofuran, but the solvent usable in the step is dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydro. Furan (THF), methylene chloride, toluene, acetonitrile and mixtures thereof can be used.

Further, in the above step, the reaction temperature is not particularly limited, but preferably, it is preferably performed at a boiling point in the solvent at -90 to -50 ° C, and the reaction time is not particularly limited, but it is preferable to react for 0.5-20 hours. .

In the method for preparing a compound represented by Chemical Formula 1 according to the present invention, after performing Step 3, optionally, as in Scheme 2 below, Step 4 may be additionally performed.

[Scheme 2]

As confirmed in Scheme 2 above,

When R ² in Chemical Formula 1 is = O, it is possible to prepare a compound in which R ² is a -OH group by performing as in step 4 above.

At this time, the reaction of step 4 is carried out a reaction to reduce the = O group of the compound represented by the formula 1 'of the reaction formula ² to prepare a compound represented by the formula 1 "R ² is -OH group. The reduction reaction is not limited thereto, but may be carried out by reacting a compound represented by Chemical Formula 1 ', Me ₄ NB (OAc) ₃ H (tetramethylammonium triacetoxyborohydride).

At this time, the solvent used in step 4 may be preferably a mixed solvent of acetic anhydride and acetonitrile anhydride, but is not limited thereto.

In addition, in the above step, the reaction temperature is not particularly limited, but is preferably performed at a boiling point in the solvent at -70 to -10 ° C, and the reaction time is not particularly limited, but it is preferable to react for 5-30 hours. .

As described above, the preparation method according to the present invention should be understood as a description for performing the preparation of the compound of the present invention. In addition, all the above-described steps may be performed as a single step or a method performed through two or more steps, and may be performed as inverting each step in consideration of a desired compound, an achievement yield, and the like. If it is possible to change or modify in the manufacturing method of the invention, it will be easily understood that it is included in the present invention. Furthermore, it should be understood that if it is a method that can be performed by common knowledge or a known technique in the meaning of being further included in the spirit of the present invention, it should be understood as being easily included in the manufacturing method of the present invention.

That is, if a person skilled in the art can easily derive by modifying or modifying the production method of the present invention in consideration of the desired compound, achievement efficiency, and the like from known techniques, the present invention is included in the present invention. Any method that can be manufactured in the same way is included in the present invention.

Furthermore, the present invention provides a pharmaceutical composition for preventing or treating acetylcholinesterase related diseases containing the compound, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Here, the acetylcholinesterase-related disease, as in the background art of the present invention described above, has been found to have a correlation with acetylcholinesterase to date, preferably inhibiting the activity of acetylcholinesterase Any disease that can be improved, improved, prevented or treated.

In addition, the compound of Formula 1 according to the present invention, its isomer, or a pharmaceutically acceptable salt, hydrate, or solvate thereof has an inhibitory activity of acetylcholinesterase itself, but after being absorbed into the body, a special in vivo environment It does not exclude the possibility of pharmacological action as an agonist or by products of metabolic processes.

At this time, the acetylcholinesterase-related disease is not limited to this, but includes all diseases that can be expressed from a phenomenon other than normal activity such as enzyme abnormality, modification, and overexpression, and in particular, for example, nervous system disease. , When the nervous system disease is due to the abnormal activity of acetylcholinesterase, the compound of the present invention, its stereoisomers, and acceptable salts thereof excellently inhibit enzymatic activity and are referred to as acetylcholinesterase related diseases. It can be useful for the improvement, prevention or treatment of symptoms or diseases.

For example, the acetylcholinesterase-related disease may be a nervous system disease, and the neurological disease may be a brain nervous system disease or a central nervous system disease.

Most preferably, the acetylcholinesterase related diseases are Alzheimer's disease, stroke, cerebral infarct, head trauma, cerebral arteriosclerosis, and Parkinson's disease disease Huntington's disease, Creutzfeldt-Jakob disease, Pick disease, Lewy body disease, Amyotrophic lateral sclerosis, multiple sclerosis and Ischemic brain disease, cerebrovascular disease, cranial nerve disease, cognitive disease or disorder, schizophrenia, attention deficit hyperactivity disorder (ADHD), attention deficit disease (ADD), central nervous system (CNS) or peripheral nervous system (PNS) ) Disease, Guillain-Barre syndrome, one or more selected from the group consisting of progressive dementia and progressive ataxia due to gradual death of nerve cells.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of a nervous system disease containing the compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

Here, the compound of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof may be characterized by improving, preventing or treating a nervous system disease by inhibiting acetylcholinesterase, and the chemical formula 1 according to the present invention Compounds, isomers thereof, or pharmaceutically acceptable salts, hydrates, or solvates thereof have an inhibitory activity of acetylcholinesterase in themselves, but after being absorbed into the body, the product may undergo metabolic processes, or by a special in vivo environment. It does not exclude the possibility of pharmacological action as an agonist. The nervous system diseases include Alzheimer's disease, stroke, cerebral infarct, head trauma, cerebral arteriosclerosis, and Parkinson's disease, Huntington's disease, Creutz Creutzfeldt-Jakob disease, Pick disease, Lewy body disease, Amyotrophic lateral sclerosis, multiple sclerosis and ischemic brain disease, Cerebrovascular disease, cranial nerve disease, cognitive disease or disorder, schizophrenia, attention deficit hyperactivity disorder (ADHD), attention deficit disease (ADD), central nervous system (CNS) or peripheral nervous system (PNS) disease, Guillain-Barre syndrome, neuro It is one or more selected from the group consisting of progressive dementia and progressive ataxia due to the gradual death of cells.

As used herein, "nervous system disease" refers to a disorder of the subject's (subject) or individual (organic) nervous system, wherein the nervous system is largely classified into the central nervous system of the brain and spinal cord, except for the peripheral nervous system do.

Diseases of the nervous system are classified as injuries, vascular disorders, infections, degeneration, structural defects, and swelling. Symptoms include chronic or acute headache, loss or tingling of the sensation, weakness and fatigue of muscles, and decreased vision, Memory loss, ataxia, muscle stiffness, and muscle cramps.

In the pharmaceutical composition according to the present invention, the compound represented by Chemical Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof may be administered in various dosage forms, oral and parenteral, during clinical administration. It can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc., which are usually used.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules, liquids, suspensions, emulsifiers, syrups, granules, elixirs, troches, etc., and these formulations include diluents (e.g., lactose) , Dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycol). Tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and, if desired, a boron such as starch, agar, alginic acid or its sodium salt, etc. It can contain an releasing or boiling mixture and / or absorbent, colorant, flavoring agent, and sweetening agent.

The pharmaceutical composition using the compound represented by Formula 1 as an active ingredient may be administered parenterally, and parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

At this time, in order to formulate a formulation for parenteral administration, a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed with water with a stabilizer or a buffer to prepare a solution or suspension, and it is administered in ampoules or vials. Can be produced. The composition may be sterile and / or contain preservatives, stabilizers, hydrating or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances. It can be formulated according to the chemical or coating method.

Examples of formulation are as follows.

<Production Example 1> Preparation of powder

Derivative represented by Formula 1: 2 g

Lactose: 1 g

A powder was prepared by mixing the above components and filling the gas-tight fabric.

<Formulation Example 2> Preparation of tablets

Derivative represented by Chemical Formula 1: 100 mg

Corn starch: 100 mg

Lactose: 100 mg

Magnesium stearate: 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional tablet manufacturing method.

<Formulation Example 3> Preparation of capsules

Derivative represented by Chemical Formula 1: 100 mg

Corn starch: 100 mg

Lactose: 100 mg

Magnesium stearate: 2 mg

After mixing the above components, the capsules were prepared by filling the gelatin capsules according to a conventional capsule preparation method.

<Formulation Example 4> Preparation of injection

Derivative represented by Chemical Formula 1: 100 mg

Mannitol: 180 mg

Na ₂ HPO ₄ ㆍ 2H ₂ O: 26 mg

Distilled water: 2974 mg

According to the conventional method of preparing an injection, the above ingredients were contained in the indicated amounts to prepare an injection.

The example of the formulation is related to the conventional formulation example, it can be easily understood by those of ordinary skill in the art that the formulation of the present invention is not limited thereto.

Furthermore, the dosage of the compound represented by Formula 1 of the present invention or a pharmaceutically acceptable salt thereof to the human body may vary depending on the age, body weight, sex, dosage form, health condition and disease degree of the subject, and the body weight Based on an adult subject of 70 Kg, it is generally 0.1-1000 mg / day, preferably 1-500 mg / day, and is divided into once to several times a day at regular time intervals according to the judgment of a doctor or pharmacist It may be administered.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, chemotherapy, and biological response modifiers for the prevention or treatment of diseases caused by Eurotensin-II receptor hyperactivity. have.

Furthermore, the present invention provides a health functional food composition for preventing or improving diseases related to acetylcholinesterase containing the compound, stereoisomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Here, the acetylcholinesterase-related disease, as in the background art of the present invention described above, has been found to have a correlation with acetylcholinesterase to date, preferably inhibiting the activity of acetylcholinesterase Any disease that can be improved, improved, prevented or treated.

At this time, the acetylcholinesterase-related disease is not limited to this, but includes all diseases that can be expressed from a phenomenon other than normal activity such as enzyme abnormality, modification, and overexpression, and in particular, for example, nervous system disease. , When the nervous system disease is due to the abnormal activity of acetylcholinesterase, the compound of the present invention, its stereoisomers, and acceptable salts thereof excellently inhibit enzymatic activity and are referred to as acetylcholinesterase related diseases. It can be useful for the improvement, prevention or treatment of symptoms or diseases.

For example, the acetylcholinesterase-related disease may be a nervous system disease, and the neurological disease may be a brain nervous system disease or a central nervous system disease.

Most preferably, the acetylcholinesterase related diseases are Alzheimer's disease, stroke, cerebral infarct, head trauma, cerebral arteriosclerosis, and Parkinson's disease disease Huntington's disease, Creutzfeldt-Jakob disease, Pick disease, Lewy body disease, Amyotrophic lateral sclerosis, multiple sclerosis and Ischemic brain disease, cerebrovascular disease, cranial nerve disease, cognitive disease or disorder, schizophrenia, attention deficit hyperactivity disorder (ADHD), attention deficit disease (ADD), central nervous system (CNS) or peripheral nervous system (PNS) ) Disease, Guillain-Barre syndrome, one or more selected from the group consisting of progressive dementia and progressive ataxia due to gradual death of nerve cells.

In addition, the present invention provides a method for treating an acetylcholinesterase related disease, comprising administering the compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof to a subject in a therapeutically effective amount.

In this method of treatment, an effective amount of one or more compounds prepared according to pharmaceutical and veterinary practice is administered to a subject to be treated, preferably a mammal, especially a human, a production animal or a livestock. The indication and performance of these treatments depends on the individual case (subject), medical evaluation taking into account existing signs, syndromes and / or dysfunctions, specific signs, risks of developing syndromes and / or dysfunctions, and other factors. (Diagnosis).

In general, treatment is carried out in single or repeated administration, and if appropriate, administered together or alternately with other active compounds or active compound-containing formulations, preferably orally at about 0.1 to 1000 mg / kg body weight, or For parenteral administration at about 0.1 to 100 mg / kg body weight, it is supplied to subjects to be treated daily.

The invention also relates to the manufacture of a pharmaceutical composition for treating a subject, preferably a mammal, in particular a human, a production animal or a livestock. Accordingly, the ligand is usually administered in the form of a pharmaceutical composition comprising a pharmaceutically acceptable excipient with one or more compounds of the invention, and, where appropriate, with other active compounds. These compositions can be administered, for example, by oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, intranasal or intracerbroventricular injection.

Examples of suitable pharmaceutical formulations are solid pharmaceutical forms, such as powders, granules, tablets, in particular capsules, lozenges, chasing agents, quesing agents, dragees, capsules, for example hard gelatin capsules and soft gelatin. Capsules, suppositories or vaginal pharmaceutical forms, semi-solid pharmaceutical forms, such as ointments, creams, hydrogels, pastes or plasters, and also liquid pharmaceutical forms, such as solvents, emulsifiers, especially oil-in-water emulsifiers, suspensions, for example For example, lotions, injections and infusions, eye drops and ear drops. Implanted release devices can also be used to administer inhibitors according to the invention. In addition, liposomes or microspheres can be used.

When preparing a formulation, the compound of the invention is optionally mixed or diluted with one or more excipients. The excipient can be a solid, semi-solid liquid material, and is used as a vehicle, carrier, or medium for the active compound.

Suitable excipients are described in the Professional Medicine Book. In addition, the formulation may be a pharmaceutically acceptable carrier or conventional auxiliary material, such as a lubricant; Wetting agents; Emulsifiers and suspending agents; Preservatives; Antioxidants; Anti-irritants; Chelating agents; Coating aids; Emulsifying stabilizers; Film formers; Gel formers; Odor blockers; Taste correctors; Suzy; Hydrocolloids; menstruum; Stabilizer; Neutralizing agent; Diffusion accelerators; Pigment; Quaternary ammonia compounds; Reattaching agents and overfatting agents; Ointment, cream or oil raw material; Silicone derivatives; Spray aids; Stabilizer; Sterilizers; Suppository base; Tablet auxiliaries, such as binders, fillers, lubricants, disintegrants or coating agents; Propellant; drier; Opacifiers; Thickeners; Wax; Plasticizers and white mineral oils. Formulations related thereto are based on, for example, expert notices described in the literature (Fiedler, HP, Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik und angrenzende Gebiete [Encyclopedia of auxiliary substances for pharmacy, cosmetics and related fields], 4th edition, Aulendorf: ECV-Editio-Kantor-Verlag, 1996].

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form may contain the following examples of inert diluents commonly used in the art: water or other solvents, solubilizers, and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate , Benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (e.g. cottonseed, peanut, corn, germ, olive, caster, and sesame oil), glycerol, tetrahydro Fatty acid esters of furfuryl alcohol, polyethylene glycol and sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents, and fragrances.

Injectable formulations, such as sterile injectable aqueous or lipid-producing suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in non-toxic parenterally acceptable diluents or solvents, for example solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used, water, Ringer's solution, U.S.P. And isotonic sodium chloride solution. In addition, sterile, fixed oil is conventionally used as a solvent or dispersion medium. Any blend fixed oil can be used for this purpose, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the manufacture of injections.

Injectable formulations can be sterilized, for example, by filtration through a bacterial-fixed filter, or by incorporating a yellowing agent into the presence of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. have.

To sustain the effectiveness of the compounds of the present invention, slow absorption of the compounds from subcutaneous or intramuscular injection is often desired. This can be achieved by the use of liquid suspensions of crystalline or amorphous substances with poor water solubility. The absorption rate of a compound depends on the crystal size and its dissolution rate, which can depend on the crystal form. Alternatively, delayed absorption of a parenterally administered compound form is achieved by dissolving or suspending the compound in an oil vehicle. Injectable formulations are made by forming microencapsule matrices of compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions compatible with body tissue.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with: at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) filler or bulking agent such as starch, lactose , Sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrants Agar--agar, calcium dehydrate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarders such as paraffins, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, Cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricant examples For talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also include a buffer.

Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules using lactose or lactose as well as excipients such as high molecular weight polyethylene glycols and the like. Tablets, dragees, capsules, pills, and granules in solid dosage form can be prepared with coatings and shells such as enteric coatings and other coatings well known in pharmaceutical formulation technology. It may optionally contain an opacifying agent, and may also be a composition that releases only the active ingredient (s), eg, in certain parts of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Similar types of solid compositions can also be used as fillers in soft and hard-filled gelatin capsules using lactose or lactose as well as excipients such as high molecular weight polyethylene glycols and the like.

The active compound can also be in micro-encapsulated form with one or more excipients as described above. Tablets, dragees, capsules, pills, and granules in solid dosage form can be prepared with coatings and shells such as enteric coatings, controlled release coatings and other coatings well known in pharmaceutical formulation technology. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms can also include, as in normal practice, additional substances other than inert diluents, such as tableting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form may also include a buffer. It may be a composition that may optionally contain an opacifying agent and also releases only the active ingredient (s), eg, in certain parts of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed with a pharmaceutically acceptable carrier and any necessary preservatives or buffers under sterile conditions, as required. It is also contemplated that ophthalmic formulations, ear drops, and eye drops are within the scope of the present invention. Additionally, the present invention contemplates the use of transdermal patches with the added advantage of providing controlled cleavage of the compound to the body. Such dosage forms can be made by dissolving or dispersing the compound in the proper medium. Absorption accelerators can also be used to increase the flow of compounds across the skin. The rate can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Additional agents may be administered separately from the combination therapy provided as part of multiple dose regimen. Alternatively, the formulations can be part of a single dosage form mixed with a compound of the invention. If administered as part of a combination therapy, the two therapeutic agents can be given simultaneously, sequentially, or intermittently. The combination therapy can be used for any of the therapeutic indications described herein.

Another aspect of the invention relates to inhibiting acetylcholinesterase activity in a biological sample or subject, the method comprising administering a compound represented by Formula 1, or a composition comprising the compound, or to the biological Contacting the sample. The term "biological sample", as used herein, generally includes in vivo, in vitro, and ex vivo substances, and also includes, without limitation, cell cultures or extracts thereof; Biopsied material obtained from a mammal or extracts thereof; And blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

Another aspect of the invention is to provide a kit comprising a separate container in a single package, wherein the compound or pharmaceutical composition disclosed herein and / or a salt thereof is one or more of which act as an acetylcholinesterase. It is provided with one or more pharmaceutically acceptable carriers for use in treating disorders, symptoms and diseases.

In the present invention, the inventor developed a compound that inhibits acetylcholinesease (acetylcholinesterase), which is known as an effective target of a nervous system disease, as described in the background of the present specification, in order to develop a drug that is meaningful for the treatment of nervous system diseases. As part of the method, a pharmacological model for acetylcholinesterase was constructed using Schrödinger's 'Phase' as part of the method, and the structure of the compound expected to be inhibited was derived (1 and 1 in the experiment below). Reference).

By performing the above-described method, the present inventors synthesized the compounds of Examples 1-8, and to evaluate the specific inhibitory activity, the inhibitory activity of each compound with respect to the acetylcholinesease enzyme was evaluated and compared. As a result, surprisingly, the compounds of Examples 1-8 of the present invention were confirmed to have an inhibitory effect of about 65% or more, based on the treatment of 10 μM of acetylculin esterase (see Experimental Example 2 and FIG. 2).

Accordingly, the compounds according to the present invention, stereoisomers thereof and pharmaceutically acceptable salts thereof can be used as target inhibitors of novel structures of acetylcholinesterase, from which neurological diseases such as brain neurological diseases, central nervous system It was found that the disease, a more specific disease, has a useful effect as a pharmaceutical composition for improving, preventing or treating Alzheimer's disease.

Hereinafter, the present invention will be described in detail by examples and experimental examples.

However, the following examples and experimental examples are only to illustrate and illustrate the present invention, but the content of the present invention is not limited thereto.

< Example  1> 1- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) -4- (9H- Carbazole Preparation of -4-yl) -4-hydroxybutan-2-one

Step 1: benzyl -[2- (2- Methoxy - Phenoxy )-ethyl]- Amine  Produce

As described above of the present invention, the target compound of step 1 could be prepared by the synthesis method of two preparation examples.

[Production Example 1]

After adding triethylamine (6051 mg, 59.80 mmol) to toluene (15 ml) in which benzyl bromide (4100 mg, 23.92 mmol) was dissolved, the temperature was lowered to 0 ° C. Toluene (10 ml) in which 2- (2-methoxyphenoxy) ethanamine (5000 mg, 29.90 mmol) was dissolved was slowly added dropwise at 0 ° C, and the temperature was increased to 25 ° C. After stirring the reaction mixture at room temperature for 8 hours, water (25 ml) was added dropwise to the reaction mixture, and stirred for 15 minutes. Then, the organic layer was separated, washed with water (15 ml), dried over sodium sulfate, and concentrated in vacuo to give a crude product. Subsequently, the crude product was purified under silica gel and 15-80% ethyl acetate / hexane conditions using column chromatography to obtain a target compound as a final group.

¹ H-NMR (600 MHz, CDCl ₃ ) δ 7.37-7.32 (m, 4H), 7.27-7.24 (m, 1H), 6.93-6.88 (m, 4H), 4.17-4.15 (t, 2H), 3.88 ( s, 2H), 3.84 (s, 3H), 3.07-3.05 (t, 2H), 1.97 (s, 1H) ppm;

[Production Example 2]

2- (2-methoxyphenoxy) ethanamine (851 mg, 5.1 mmol), benzaldehyde (540 mg, 5.1 mmol), para-toluenesulfonic acid (PTSA, p-Toluenesulfonic acid) (105 mg, 0.55 mmol) Was dissolved in 5 mL of toluene. The mixture was refluxed to maintain it, and stirred. After confirming the completion of the reaction using TLC (Thin Layer Chromatography), the temperature was lowered to room temperature. The mixed solution was separated using water and ethyl acetate, and washed with saturated aqueous sodium chloride solution. The organic layer was dried over magnesium sulfate and concentrated in vacuo to yield the crude product.

The crude product prepared from the above was again dissolved in methanol (10 ml) at 0 ° C. Then, sodium borohydride (250 mg, 6.6 mmol) was slowly added dropwise, and the temperature was raised to room temperature. After the reaction for 24 hours, the mixture was diluted with dichloromethane and water. The diluted solution was extracted three times with saturated aqueous sodium chloride solution, separated and washed. Then, dried over magnesium sulfate, concentrated in vacuo, and purified using flash column chromatography under silica gel, 10-50% ethyl acetate / hexane conditions, to finally obtain the target compound.

¹ H-NMR (600 MHz, CDCl ₃ ) δ 7.37-7.32 (m, 4H), 7.27-7.24 (m, 1H), 6.93-6.88 (m, 4H), 4.17-4.15 (t, 2H), 3.88 ( s, 2H), 3.84 (s, 3H), 3.07-3.05 (t, 2H), 1.97 (s, 1H) ppm;

step 2: 1 -( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) Preparation of propan-2-one

Dissolve the compound (2400 mg, 9.32 mmol) and 1-chloropropan-2-one (1790 mg, 18.74 mmol) prepared in step 1 in tetrahydrofuran (15 ml) containing triethylamine at room temperature. After stirring for 15 hours, it was filtered. The filtrate was then diluted with ethyl acetate (80 ml), and washed with saturated aqueous sodium hydrogen carbonate solution and aqueous sodium chloride solution. Then dried over anhydrous magnesium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography under silica gel, 15-80% ethyl acetate / hexane conditions, and finally a yellow target compound was obtained.

¹ H-NMR (600 MHz, CDCl ₃ ) δ 7.38-7.37 (m, 2H), 7.33-7.31 (m, 2H), 7.27-7.25 (m, 1H), 6.94-6.84 (m, 4H), 4.12- 4.11 (t, 2H), 3.84 (s, 3H), 3.81 (s, 2H), 3.06-3.04 (t, 2H), 2.12 (s, 3H) ppm;

step 3: 1 -( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) -4- (9H- Carbazole Preparation of -4-yl) -4-hydroxybutan-2-one

First, a tetrahydrofuran solution (133.4 mg, 1.24 mmol) of 1 mol concentration of lithium diisopropylamide (LDA) was lowered to -78 ° C for 10 minutes. To this, the compound prepared in step 2 (300 mg, 0.95 mmol) was dissolved in tetrahydrofuran (1.5 ml), and then slowly added dropwise. After the mixture was stirred for 20 minutes, 9H-carbazole-4-carbaldehyde (187.0 mg, 0.95 mmol) was quickly added dropwise thereto, followed by stirring for 1 hour. The reactivity was reduced using a diluted ammonium chloride solution (pH 4-5), and the organic layer was separated using ethyl acetate and water. The separated organic layer was washed again with ammonium chloride solution. Subsequently, silica gel inactivated with ammonia water, purified under 1-15% ethyl acetate / hexane conditions using column chromatography, dried under sodium sulfate and vacuum conditions, and finally obtained the target compound of Example 1.

¹ H-NMR (600 MHz, Acetone-d ₆ ) δ 10.45 (s, 1H), 8.28-8.27 (d, J = 8.28 Hz, 1H), 7.53-7.52 (d, 1H), 7.43-7.36 (m, 6H), 7.28-7.26 (m, 2H), 7.22-7.21 (m, 1H), 7.18-7.15 (m, 1H), 6.88-6.81 (m, 4H), 6.11-6.10 (d, J = 9.6 Hz, 1H), 4.09-4.08 (m, 2H), 3.90-3.83 (m, 2H), 3.67 (s, 3H), 3.17-3.13 (m, 1H), 3.03-3.01 (t, 2H), 2.92-2.88 ( dd, J = 2.92 Hz, J = 2.89 Hz, 1H), 2.84-2.76 (m, 3H) ppm.

< Example  2> 1- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) -4-hydroxy-4- (9- methyl Preparation of -9H-carbazol-4-yl) butan-2-one

In place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 1, except that 9-methyl-9H-carbazole-4-carbaldehyde was used, it was carried out in the same manner as in Example 1 above. The desired compound was prepared.

¹ H-NMR (600 MHz, Acetone-d ₆ ) δ 8.30-8.29 (d, J = 8.00 Hz, 1H), 7.56-7.54 (d, J = 7.55 Hz, 1H) 7.47-7.45 (m, 4H), 7.42-7.41 (m, 2H), 7.28-7.25 (m, 2H), 7.22-7.19 (m, 2H), 6.90-6.79 (m, 4H), 6.14-6.12 (dd, J = 2.5 Hz, J = 2.5 Hz, 1H), 4.08-4.06 (m, 2H), 3.93-3.91 (s, 3H), 3.85-3.78 (m, 4H), 3.67-3.66 (s, 3H), 3.65 (s, 1H), 3.17- 3.11 (dd, J = 9.8 Hz, J = 9.8 Hz, 1H), 3.02-3.00 (m, 2H), 2.93-2.87 (dd, J = 2.8 Hz, J = 2.8 Hz, 1H) ppm.

< Example  3> 1- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino )-4-( Biphenyl Preparation of -3-yl) -4-hydroxybutan-2-one

In the same manner as in Example 1, except that [1,1'-biphenyl] -3-carbaldehyde was used in place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 1 above. The desired compound was prepared.

¹ H-NMR (600 MHz, Acetone-d) δ 7.65-7.68 (m, 3H), 7.53-7.55 (m, 1H), 7.38-7.49 (m, 7H), 7.31-7.34 (m, 2H), 7.24 -7.28 (m, 1H), 6.84-6.94 (m, 4H), 5.24 (dd, J = 8.7 Hz, J = 3.9 Hz, 1H), 4.47 (s, 1H), 4.07-4.13 (m, 2H), 3.82-3.90 (m, 3H), 3.76 (s, 3H), 3.55-3.64 (m, 2H), 2.99-3.02 (m, 2H), 2.91 (dd, J = 15.6 Hz, J = 16.2 Hz, J = 4.2 Hz, 1H) ppm.

< Example  4> 1- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino )-4-( Biphenyl Preparation of -4-yl) -4-hydroxybutan-2-one

In the same manner as in Example 1, except that [1,1'-biphenyl] -4-carbaldehyde was used in place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 1 above. The desired compound was prepared.

¹ H-NMR (600 MHz, CDCl ₃ ) δ 7.58-7.53 (m, 4H), 7.45-7.42 (m, 2H), 7.36-7.26 (m, 8H), 6.95-6.84 (m, 4H), 5.12 ( dd, J = 3.7 Hz, J = 3.6 Hz, 1H), 4.14-4.12 (m, 2H), 3.83-3.81 (m, 2H), 3.77 (s, 3H), 3.53 (s, 2H), 3.07-3.05 (m, 2H), 2.95-2.87 (m, 2H) ppm.

< Example  5> 4- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) -1- (9H- Carbazole -4-yl) Preparation of butane-1,3-diol

After performing the same manner as in Example 1 to prepare a compound, it was performed as in Preparation Example 3 to prepare a target compound.

[Production Example 3]

First, tetramethylammonium triacetoxyborohydride (206 mg, 0.08 mmol) was added to a mixed solvent of acetic anhydride (0.5 mL) and acetonitrile anhydride (0.5 mL), and stirred at -50 ° C under argon gas. The solution (50 mg, 0.01 mmol) prepared in Example 1 was dissolved in anhydrous acetonitrile (0.2 mL), the temperature was raised to -20 ° C, and the mixture was stirred for 20 hours. The reactivity was reduced with potassium tartrate and extracted three times with methylene chloride. The organic layer was dehydrated with anhydrous sodium sulfate, and concentrated in vacuo to give a crude product. The crude product was purified by flash column chromatography under silica gel, 15-50% ethyl acetate / hexane conditions, to finally obtain the target compound.

¹ H-NMR (600 MHz, Acetone-d) δ 10.24 (s, 1H), 8.36 (d, J = 7.8 Hz, 1H), 7.32-7.37 (m, 2H), 7.21-7.27 (m, 5H), 7.06-7.14 (m, 3H), 6.96-6.99 (m, 1H), 6.71-6.82 (m, 4H), 5.85 (d, J = 8.4 Hz, 1H), 4.18-4.22 (m, 1H), 3.90- 3.95 (m, 3H), 3.67 (s, 3H), 3.65-3.78 (m, 2H), 3.59 (d, J = 13.8 Hz, 1H), 2.87-2.91 (m, 1H), 2.73-2.77 (m, 1H, shadowed with H2O), 2.49-2.55 (m, 2H) 1.82-1.86 (m, 1H, shadowed with H2O), 1.72-1.76 (m, 1H) ppm

< Example  6> 4- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino ) -1- (9- methyl -9H- Carbazole -4-yl) Preparation of butane-1,3-diol

In place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 5, it was carried out in the same manner as in Example 5, except that 9-methyl-9H-carbazole-4-carbaldehyde was used. The desired compound was prepared.

¹ H-NMR (600 MHz, Acetone-d ₆ ) δ 8.53-8.51 (d, J = 8.0 Hz, 1H), 7.53-7.50 (m, 2H), 7.46-7.44 (m, 2H), 7.41-7.38 ( m, 3H), 7.27-7.25 (m, 2H), 7.21-7.19 (m, 1H), 7.16-7.13 (m, 1H), 6.95-6.94 (m, 1H), 6.91-6.83 (m, 3H), 6.01-5.99 (dd, J = 2.0 Hz, J = 2.1 Hz, 1H), 4.35-4.31 (m, 1H), 4.06-4.04 (m, 2H), 3.89 (s, 3H), 3.80 (s, 3H) , 3.74-3.71 (d, 1H), 3.04-3.00 (m, 1H), 2.89-2.85 (m, 2H), 2.66-2.64 (m, 2H), 1.99-1.95 (m, 1H), 1.89-1.84 ( m, 1H) ppm.

< Example  7> 4- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino )-One-( Biphenyl -3-yl) Preparation of butane-1,3-diol

In the same manner as in Example 5, except that [1,1'-biphenyl] -3-carbaldehyde was used in place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 5 above. The desired compound was prepared.

¹ H-NMR (600 MHz, Acetone-d) δ 7.50-7.54 (m, 3H), 7.35-7.37 (m, 1H), 7.21-7.32 (m, 8H), 7.13-7.16 (m, 1H), 7.06 -7.09 (m, 1H), 6.70-6.82 (m, 4H), 4.92 (dd, J = 9.3 Hz, J = 2.7 Hz, 1H), 4.41-4.62 (m, 2H), 3.95-3.97 (m, 2H ), 3.67 (s, 3H), 3.33-3.48 (m, 2H), 2.85-2.90 (m, 1H), 2.75-2.79 (m, 1H), 2.50-2.57 (m, 2H), 1.72-1.77 (m , 1H), 1.59-1.64 (m, 1H) ppm.

< Example  8> 4- ( Benzyl (2- (2-methoxyphenoxy) ethyl) amino )-One-( Biphenyl -4-yl) Preparation of butane-1,3-diol

In the same manner as in Example 5, except that [1,1'-biphenyl] -4-carbaldehyde was used in place of 9H-carbazole-4-carbaldehyde used in Step 3 of Example 5 above. The desired compound was prepared.

The structural formulas of the compounds prepared in Examples 1-8 are shown in Table 2 below.

Example constitutional formula Example constitutional formula One

5

2

6

3

7

4

8

< Experimental example  1> Acetylcholine esterase Pharmacist  Model construction and predictive evaluation of new inhibitory compounds using the same

In the present invention, in order to predict a new acetylcholinesterase compound, experiments were performed in the following manner.

Specifically, in order to identify a novel inhibitory compound for acetylcholinesterase, which is an effective target of a nervous system disease, a pharmacophore model represented by FIG. 1 is used using Schrödinger's pharmacological modeling program "Phase". Was constructed, and virtual search was performed using the produced pharmacological model, and more specifically, by predicting the acetylcholinesterase inhibitory efficacy of the compound represented by Formula 1 of the present invention based on the input data to the library, finally The compound represented by Chemical Formula 1 of the present invention was derived as a candidate group for a virtual active material.

Figure 1 shows a model of the acetylcholinesterase pharmacological group, manufactured with Schrodinger's "Phase" f, to derive the structure of a novel acetylcholinesterase inhibitory compound.

From Experimental Example 1 and FIG. 1, it can be seen that the compound represented by Formula 1 of the present invention derived has a useful effect as a novel compound that inhibits acetylcholinesterase, and furthermore, from the inhibitory effect of acetylcholinesterase. It can be seen that it can be used as an active ingredient of a nervous system disease that can be improved, prevented or treated.

< Experimental example  2> Acetylcholine esterase  Inhibition activity evaluation

In order to evaluate the inhibitory activity of the example compound prepared according to the present invention for acetylcholinesterase, it was tested as follows.

Specifically, the acetylcholine degrading enzyme (eeAChE, EC 3.1.1.7), acetylcholine chloride (ACh), and acetylcholine chloride (ACh quantification colorimetric assay kit from BioVision) of the electric eel (Electrophorus electricus) After being prepared for this measurement, in order to measure the enzyme inhibitory ability of the example compound, experiments were performed in the following order based on the specified specification method of the Ach kit manufacturer (Draczkowski, P. et al. Bba-Gen Subjects 1860 , 967-974, doi: 10.1016 / j.bbagen.2015.11.006 (2016)).

First, acetylcholine degrading enzyme (eeAChE, EC 3.1.1.7) was dissolved in 20 μM Tris-HCL buffer, and the concentration of acetylcholine degrading enzyme was prepared to be 5 μM. Acetylcholine chloride was dissolved in 20 mM Tris-HCL buffer to prepare an aqueous solution of Tris-HCL buffer at a concentration of 10 μM acetylcholine chloride. Example compounds were prepared in two concentrations of 10 μM and 20 μM. The prepared acetylcholine chloride solution and the example solution were mixed with 20 μM acetylcholine degrading enzyme. After mixing, measurement of the inhibitory efficacy of the acetylcholine degrading agent (Example compound), which decomposes acetylcholine to choline at 20 and 30 minutes, was evaluated by measuring the color change of the quantitative kit at a wavelength of 570 nm. The results of all experiments were conducted twice or more to derive the mean and standard deviation, and the results are shown in Table 3 and FIG. 2 below.

Example After the experiment, the residual activity of acetylcholinesterase (%) 10 μM 20 μM Untreated 100 (reference active) One 150 51 2 82 53 3 57 40 5 68 39 6 51 35 7 52 35

Figure 2 is a graph showing the acetylcholinesterase inhibitory activity of the compound of the present invention,

As can be seen from Table 3 and Figure 2, it can be seen that the activity of the acetylcholine esterase is excellently inhibited to at least 65% or more from the treatment of the example compound.

Therefore, the compound according to the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, can excellently inhibit the activity of acetylcholinesterase, including it as an active ingredient, a nervous system disease, preferably a brain nervous system It can be useful as a pharmaceutical composition for the prevention or treatment of diseases, central nervous system diseases.

Claims (15)

A compound represented by Formula 1 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In the above formula 1,
R ¹ is unsubstituted or substituted benzyl,
Here, the substituted benzyl is substituted with one or more substituents selected from the group consisting of halogen and C _1-3 linear or branched alkyl;

R ² is = O or -OH; And

R ³ is unsubstituted or substituted biphenyl or unsubstituted or substituted carbazole,
Here, the substituted biphenyl or substituted carbazolyl is substituted with one or more substituents selected from the group consisting of halogen and C _1-3 linear or branched alkyl).
delete According to claim 1,
R ³ is

,

,

, or

A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
According to claim 1,
The compound represented by Formula 1 is a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, characterized in that it is any one selected from the following group of compounds:
(1) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (9H-carbazol-4-yl) -4-hydroxybutan-2-one;
(2) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4-hydroxy-4- (9-methyl-9H-carbazol-4-yl) butan-2-one;
(3) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (biphenyl-3-yl) -4-hydroxybutan-2-one;
(4) 1- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -4- (biphenyl-4-yl) -4-hydroxybutan-2-one;
(5) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (9H-carbazol-4-yl) butane-1,3-diol;
(6) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (9-methyl-9H-carbazol-4-yl) butane-1,3-diol;
(7) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (biphenyl-3-yl) butane-1,3-diol; And
(8) 4- (benzyl (2- (2-methoxyphenoxy) ethyl) amino) -1- (biphenyl-4-yl) butane-1,3-diol.
According to claim 1,
The compound represented by Chemical Formula 1 is a compound characterized by being derived using a pharmacophore model for acetylcholinesterase, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
delete delete As shown in Scheme 1 below,
Preparing a compound represented by Chemical Formula 3 from a compound represented by Chemical Formula 2 (step 1);
Preparing a compound represented by Formula 4 from the compound represented by Formula 3 prepared in Step 1 (Step 2); And
Preparing a compound represented by Formula 1 from the compound represented by Formula (4) prepared in Step 2 (Step 3); Method for producing a compound represented by Formula 1 of claim 1 comprising:
[Scheme 1]

(In Scheme 1 above,
R ¹ , R ² , and R ³ are as defined in Formula 1 of claim 1).
delete A pharmaceutical composition for the prevention or treatment of diseases of the nervous system, comprising the compound of claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 10,
The nervous system disease is a pharmaceutical composition characterized in that the brain nervous system disease or central nervous system disease.
The method of claim 10,
The nervous system diseases include Alzheimer's disease, stroke, cerebral infarct, head trauma, cerebral arteriosclerosis, and Parkinson's disease, Huntington's disease, Creutz Creutzfeldt-Jakob disease, Pick disease, Lewy body disease, Amyotrophic lateral sclerosis, multiple sclerosis and ischemic brain disease, Cerebrovascular disease, cranial nerve disease, cognitive disease or disorder, schizophrenia, attention deficit hyperactivity disorder (ADHD), attention deficit disease (ADD), central nervous system (CNS) or peripheral nervous system (PNS) disease, Guillain-Barre syndrome, neuro A pharmaceutical composition comprising at least one member selected from the group consisting of progressive dementia and progressive ataxia due to gradual death of cells.
delete A composition for the prevention or improvement of a nervous system disease, comprising the compound of claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.
The method of claim 14,
The nervous system disease is a functional brain food composition characterized in that the brain nervous system disease or central nervous system disease.

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