KR20240076709A - Diagnostic composition for diagnosing neurodegenerative disease and method for providing information for diagnosing the same - Google Patents

Abstract

A novel pyrrolopyridine derivative compound, a composition for diagnosing neurodegenerative diseases containing the same as an active ingredient, and a method of providing information for diagnosing neurodegenerative diseases using the composition, wherein the novel pyrrolopyridine derivative compounds are tau Since it has excellent selectivity and affinity with the (Tau) protein, it can be useful for early detection of degenerative brain diseases such as Alzheimer's disease when diagnosing neurodegenerative diseases using a composition for diagnosing neurodegenerative diseases containing it.

Description

Diagnostic composition for diagnosing neurodegenerative disease and method for providing information for diagnosing the same {Diagnostic composition for diagnosing neurodegenerative disease and method for providing information for diagnosing the same}

It relates to a novel pyrrolopyridine derivative compound, a composition for diagnosing neurodegenerative diseases containing the same as an active ingredient, and a method of providing information for diagnosing neurodegenerative diseases using the composition.

As the aging phenomenon due to the recent increase in average lifespan has emerged as a social problem, the importance of research on degenerative brain disease is being emphasized along with the increase in the elderly population. Degenerative brain diseases are known to be caused by neurodegeneration due to aging and the death of nerve cells due to protein aggregation due to genetic or environmental factors. Degenerative brain diseases cause disorders in motor function, memory, and cognition. Degenerative brain diseases include Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, Lou Gehrig's disease, etc., considering the main symptoms and brain areas affected.

Diagnosis of such dementia is currently conducted through cognitive tests and morphological changes using MR imaging, and in current nuclear medicine tests, beta-amyloid or tau is used to measure physiological and pathological changes within the brain. ) Radiopharmaceuticals for protein diagnosis are used clinically. Currently known representative radiopharmaceuticals for diagnosing dementia include six compounds, five of which are labeled F-18 and the remaining one is labeled C-11. The two compounds currently undergoing the most clinical research on tau protein are Lilly's AV-1451 (T807) and GE's THK5351, both of which are compounds labeled with the radioisotope F-18.

THK5351 AV-1451 MK-6240 PI2620 PI2620 C-11 PBB3

Due to the nature of radiopharmaceuticals, there are many restrictions in producing and using pharmaceuticals depending on the half-life of the labeled radioisotope. When using PET radioisotope (I-124), which has a long half-life, there are no time constraints on production and distribution and clinical use, and it has the advantage of being able to be supplied to each hospital from a GMP manufacturing plant that produces radiopharmaceuticals.

nuclide half life Collapse type (%) Gamma rays (MeV) production method video equipment F-18 110 minutes b+(97) 0.511 ¹⁸ O(p,n) ¹⁸ F PET I-124 April 18th b+(25.6), EC 74.4% 0.511 ¹²⁵ Te(p,2n) ¹²⁴ I PET I-123 13.2 hours EC 100% 0.159 ¹²⁴ Xe(p,2n) ¹²³ I SPECT

However, in the case of Alzheimer's disease, a representative degenerative disease, the development of compounds for diagnosing Tau protein, known as the causative agent, and compositions containing it as an active ingredient are currently in the early stages and are urgently needed.

Republic of Korea Patent Publication No. 10-2344437

The purpose of solving the above problem is as follows.

Degenerative diseases such as Alzheimer's disease For the diagnosis and early detection of diseases, a pyrrolopyridine derivative compound that is highly reactive and selective to the Tau protein, a diagnostic composition containing the same as an active ingredient, and a radioactive substance with a long half-life using the same The purpose is to provide a method of providing information for diagnosing neurodegenerative diseases using radioactive compounds using the isotope I-124.

The compound according to one embodiment, its stereoisomer, or its pharmaceutically acceptable salt is represented by the following formula (1).

[Formula 1]

R ¹ and R ² are each independently hydrogen, unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, or cyano group. (-CN);

When R ¹ and R ² are connected to form a ring, they are an unsubstituted or substituted homoaryl group of 5 or 10 atoms, or an unsubstituted or substituted 5 or 6 atom optionally containing N, S and/or O. It is a heteroaryl group of atoms;

R ³ is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, unsubstituted or substituted C _1-6 linear or branched alkoxy, unsubstituted or substituted C _6-10 aryl group, or unsubstituted or substituted 5- or 6-membered heteroaryl group optionally containing N, S and/or O. ego; and

The substituted substituent is substituted with one or more selected from the group consisting of halogen, hydroxy, C _1-5 straight or branched alkyl, unsubstituted or C 1-5 straight or branched alkoxy substituted with one or more halogens. .

R ¹ and R ² are connected to form an unsubstituted or substituted homoaryl group of 5 or 10 atoms; R ³ is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C _1-6 straight or branched alkyl, or unsubstituted or substituted C _1-6 straight or branched alkoxy; And the substituted substituent is one selected from the group consisting of halogen, hydroxy group (OH), C _1-5 straight or branched alkyl, unsubstituted or C _1-5 straight or branched alkoxy substituted with one or more halogens. It can be replaced with the above.

A compound according to another embodiment, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is represented by the following formula (2).

[Formula 2]

wherein R ^a1 to R ^a4 are each independently hydrogen or a halogen group;

R ^b is hydrogen, halogen, hydroxy group (OH), or unsubstituted or substituted C _1-6 straight or branched alkoxy; and

The substituted substituents are at least one selected from the group consisting of halogen, hydroxy group (OH), C _1-5 straight or branched alkyl, unsubstituted or C _1-5 straight or branched alkoxy substituted with one or more halogens. is replaced with

Wherein R ^a1 , R ^a2 , and R ^a4 are hydrogen, R ^a2 is a halogen group; And R ^b may be hydrogen.

The halogen group may be one selected from the group consisting of F, Cl, Br, and I.

The halogen group may be one selected from the group consisting of isotopically labeled ¹⁸ F, ⁸² Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I, and ¹³¹ I.

A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to another embodiment is represented by the following formula (3).

[Formula 3]

Said R is ¹²³ I, ¹²⁴ I or ¹²⁵ I.

A composition for diagnosing neurodegenerative diseases according to another embodiment contains the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound represented by Formula 1, its stereoisomer, or its pharmaceutically acceptable salt can bind to Tau protein.

The neurodegenerative disease may be a degenerative brain disease.

The neurodegenerative diseases include Alzheimer's disease, Parkinson's disease (PD), Parkinson's disease dementia (PDD), Lewy body dementia (DLB), Lewy body variant of Alzheimer's disease (LBVAD), multiple system atrophy (MSA), Pick disease, and tau. It may be one or more diseases selected from the group consisting of diseases.

According to another aspect, a method of providing information for diagnosing a neurodegenerative disease includes administering the composition for diagnosing a neurodegenerative disease of claim 8 to a subject; And measuring the signal for Tau protein.

The signal measurement step is performed using one or more methods selected from the group consisting of single photon emission computed tomography (SPECT), positron emission tomography (PET), magnetic resonance imaging, and autoradiography. can be measured and imaged.

According to one aspect, the pyrrolopyridine derivative compound has excellent selectivity and affinity with the Tau protein, so a composition for diagnosing neurodegenerative diseases containing it can be used for early detection of degenerative brain diseases such as Alzheimer's disease when diagnosing neurodegenerative diseases. It has an effect that can be useful.

Figure 1a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²⁵ I]KR201 (Example 1) pyrrolopyridine derivative compound, and Figure 1b is a graph showing the values obtained by HPLC purification after measuring the Radio TLC values of Figure 1a. This is the graph shown.
Figure 2a is a graph showing the RI peak after HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²⁵ I]KR201 (Example 1), and Figure 2b is a graph showing the HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²⁵ I]KR201 (Example 1). This is a graph showing the UV peak after.
Figure 3a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²⁵ I]6-MK (Comparative Example 1) pyrrolopyridine derivative compound, and Figure 3b is a graph obtained by HPLC purification after measuring the Radio TLC values of Figure 3a. This is a graph showing the values.
Figure 4a is a graph showing the RI peak after HPLC analysis of the [ ¹²⁵ I]6-MK (Comparative Example 1) pyrrolopyridine derivative compound, and Figure 4b is a graph showing the [ ¹²⁵ I]6-MK (Comparative Example 1) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis of the compound.
Figure 5a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²³ I]KR201 (Example 2) pyrrolopyridine derivative compound, and Figure 5b is a graph showing the values obtained by HPLC purification after measuring the Radio TLC values of Figure 5a. This is the graph shown.
Figure 6a is a graph showing the RI peak after HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²³ I]KR201 (Example 2), and Figure 6b is a graph showing the HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²³ I]KR201 (Example 2). This is a graph showing the UV peak after.
Figure 7a is a graph showing the Radio TLC values measured 30 minutes after preparing the [ ¹²³ I]6-MK (Comparative Example 2) pyrrolopyridine derivative compound, and Figure 7b is a graph obtained by HPLC purification after measuring the Radio TLC values of Figure 7a. This is a graph showing the values.
Figure 8a is a graph showing the RI peak after HPLC analysis of the [ ¹²³ I]6-MK (Comparative Example 2) pyrrolopyridine derivative compound, and Figure 8b is a graph showing the [ ¹²³ I]6-MK (Comparative Example 2) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis of the compound.
Figure 9a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²⁴ I]KR201 (Example 3) pyrrolopyridine derivative compound, and Figure 9b is a graph showing the values obtained by HPLC purification after measuring the Radio TLC values of Figure 9a. Figure 9c is a graph showing the UV peak after HPLC analysis of the [ ¹²⁴ I]KR201 (Example 3) pyrrolopyridine derivative compound.
Figure 10 shows the results of confirming the target-binding ability of Tau protein through autoradiography of the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK). This is an image showing .
Figure 11 shows the target-binding ability of Tau protein through immunohistochemistry of the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I] KR201) and Comparative Example 1 ([ ¹²⁵ I] 6-MK). This is a graph comparing the resulting radioactivity concentrations.
Figure 12 is a graph showing the results of radioactivity distribution in each organ over time after administering the pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I]KR201) to animals.
Figure 13 is a graph showing the results of radioactivity distribution in each organ over time after administering the pyrrolopyridine derivative compound according to Comparative Example 1 ([ ¹²⁵ I]6-MK) to animals.
Figure 14 is an image showing the results of positron emission tomography (PET) imaging over time after administering the pyrrolopyridine derivative compound according to Example 3 ([ ¹²⁴ I] KR201) to normal animals.
Figure 15 is an image showing the results of positron emission tomography (PET) imaging over time after administering the pyrrolopyridine derivative compound according to Comparative Example 3 ([ ¹²⁴ I]6-MK) to normal animals.
Figure 16 is an image showing the results of positron emission tomography (PET) imaging over time after administering the pyrrolopyridine derivative compound according to Comparative Example 4 ([ ¹⁸ F]AV-1451) to normal animals.
Figure 17 is an image showing the results of positron emission tomography (PET) imaging over time after administering the pyrrolopyridine derivative compound according to Example 3 (transgenic mouse [ ¹²⁴ I]KR201 for over 12 weeks) to transgenic animals. all.
Figure 18 shows the results of positron emission tomography (PET) imaging over time after administering the pyrrolopyridine derivative compound according to Comparative Example 4 (15-week transgenic mouse [ ¹⁸ F]AV-1451) to the transgenic animal. It's an image.

The above objectives, other objectives, features and advantages will be easily understood through the following preferred embodiments in conjunction with the accompanying drawings. However, it is not limited to the embodiments described here and may be embodied in other forms. Rather, the embodiments introduced herein are provided so that the disclosed content will be thorough and complete and the technical ideas can be sufficiently conveyed to those skilled in the art.

While describing each drawing, similar reference numerals are used for similar components. In the attached drawings, the dimensions of the structures are enlarged from the actual size for clarity of the present invention. Terms such as first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The above terms are used only for the purpose of distinguishing one component from another. For example, a first component may be named a second component without departing from the scope of the present invention, and similarly, the second component may also be named a first component. Singular expressions include plural expressions unless the context clearly dictates otherwise.

In this specification, terms such as “comprise” or “have” are intended to designate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, but are not intended to indicate the presence of one or more other features. It should be understood that this does not exclude in advance the possibility of the existence or addition of elements, numbers, steps, operations, components, parts, or combinations thereof.

Unless otherwise specified, all numbers, values, and/or expressions used herein expressing quantities of components, reaction conditions, polymer compositions, and formulations are intended to represent, among other things, how such numbers inherently occur in obtaining such values. Since they are approximations reflecting the various uncertainties of measurement, they should be understood in all cases as being qualified by the term "approximately". Additionally, where a numerical range is disclosed herein, such range is continuous and, unless otherwise indicated, includes all values from the minimum to the maximum of such range inclusively. Furthermore, when such range refers to an integer, all integers from the minimum value up to and including the maximum value are included, unless otherwise indicated.

In this specification, when a range is stated for a variable, the variable will be understood to include all values within the stated range, including the stated endpoints of the range. For example, the range "5 to 10" includes the values 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood that it also includes any values between integers that fall within the scope of the stated range, such as 5.5, 6.5, 7.5, 5.5 to 8.5, and 6.5 to 9, etc. Also, for example, the range "10% to 30%" includes values such as 10%, 11%, 12%, 13%, etc. and all integers up to and including 30%, as well as 10% to 15%, 12% to 12%, etc. It will be understood that it includes any subranges, such as 18%, 20% to 30%, etc., and any value between reasonable integers within the range of the stated range, such as 10.5%, 15.5%, 25.5%, etc.

In the case of Alzheimer's disease, a typical degenerative disease, the development of a compound for diagnosing Tau protein, known as the causative agent, and a composition containing it as an active ingredient is currently in the early stages, and development is urgently needed.

Accordingly, as a result of intensive research to solve the above problem, the present inventors discovered that a pyrrolopyridine derivative compound with a novel structure has excellent selectivity and affinity with the Tau protein, and was able to treat neurodegenerative diseases including it. The present invention was completed after discovering that the use of a diagnostic composition can be useful for early detection of degenerative brain diseases such as Alzheimer's disease.

Specifically, we produced a pyrrolopyridine derivative compound with good selectivity for the Tau protein and produced a radioactive compound using radioactive iodine (I-123, I-124, I-125), suggesting the possibility of non-clinical use. , the F-18 labeled compound, which is currently used in clinical practice, was applied to a non-clinical model to evaluate the superiority of the I-124 labeled compound.

A pyrrolopyridine derivative compound having a novel structure according to one embodiment is a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

X is CH, O, or S,

R ¹ and R ² are each independently hydrogen, unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, or cyano group. (-CN);

When R ¹ and R ² are connected to form a ring, they are an unsubstituted or substituted homoaryl group of 5 or 10 atoms, or an unsubstituted or substituted 5 or 6 atom optionally containing N, S and/or O. It is a heteroaryl group of atoms;

R ³ is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, unsubstituted or substituted C _1-6 linear or branched alkoxy, unsubstituted or substituted C _6-10 aryl group, or unsubstituted or substituted 5- or 6-membered heteroaryl group optionally containing N, S and/or O. ego; and

The substituted substituent is substituted with one or more selected from the group consisting of halogen, hydroxy, C _1-5 straight or branched alkyl, unsubstituted or C 1-5 straight or branched alkoxy substituted with one or more halogens. .

Preferably, X is CH;

R1 and R2 are connected to form an unsubstituted or substituted homoaryl group of 5 or 10 atoms;

R3 is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C1-6 straight or branched alkyl, or unsubstituted or substituted C1-6 straight or branched alkoxy; and

The substituted substituent is substituted with one or more selected from the group consisting of halogen, hydroxy group (OH), C1-5 straight or branched alkyl, unsubstituted or C1-5 straight or branched alkoxy substituted with one or more halogens. It may have happened.

The pyrrolopyridine derivative compound according to another embodiment may be a compound represented by the following formula (2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 2]

wherein R ^a1 to R ^a4 are each independently hydrogen or a halogen group;

R ^b is hydrogen, halogen, hydroxy group (OH), or unsubstituted or substituted C _1-6 straight or branched alkoxy; and

The substituted substituents are at least one selected from the group consisting of halogen, hydroxy group (OH), C _1-5 straight or branched alkyl, unsubstituted or C _1-5 straight or branched alkoxy substituted with one or more halogens. It may have been replaced with .

Preferably, R ^a1 , R ^a2 , and R ^a4 are hydrogen, and R ^a2 is a halogen group; and

The R ^b may be hydrogen.

At this time, the halogen group may be one selected from the group consisting of F, Cl, Br, and I, preferably isotopically labeled ¹⁸ F, ⁸² Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, It may be one selected from the group consisting of ¹²⁵ I, and ¹³¹ I.

The pyrrolopyridine derivative compound according to another embodiment may be a compound represented by the following formula (3), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 3]

Said R is ¹²³ I, ¹²⁴ I or ¹²⁵ I.

The compounds represented by Formula 1, Formula 2, or Formula 3 can be used in the form of pharmaceutically acceptable salts, and acid addition salts formed by pharmaceutically acceptable free acids are useful as salts. .

Specifically, acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, and hydroxyalkanoates. Non-toxic organic acids such as ates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids, trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid. Obtained from organic acids such as These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and nitrate. Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube. Latex, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, beta-hydroxybutyrate, Includes glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, etc.

The acid addition salt can be prepared by conventional methods. For example, a derivative of Formula 1, Formula 2, or Formula 3 is dissolved in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., and then added with an organic acid or inorganic acid. It can be prepared by filtering and drying the resulting precipitate, or by distilling the solvent and excess acid under reduced pressure, drying it, and crystallizing it in an organic solvent.

Additionally, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of 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 appropriate to prepare sodium, potassium or calcium salts as metal salts. Additionally, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).

Furthermore, it includes not only the compounds represented by Formula 1, Formula 2, or Formula 3 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, etc. that can be prepared therefrom.

The term “hydrate” refers to a compound of the present invention containing a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. or its salt. The hydrate of the compound represented by Formula 1, Formula 2, or Formula 3 may contain a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. The hydrate may contain more than 1 equivalent of water, preferably 1 to 5 equivalents of water. Such hydrates can be prepared by crystallizing the compounds represented by Formula 1, Formula 2, or Formula 3 of the present invention, isomers thereof, or pharmaceutically acceptable salts thereof from water or a solvent containing water.

The term “solvate” refers to a compound of the invention or a salt thereof containing a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Preferred solvents therefor are solvents that are volatile, non-toxic, and/or suitable for administration to humans.

The term “isomer” refers to a compound of the present invention or a salt thereof that has the same chemical or molecular formula but is structurally or sterically different. These isomers include structural isomers such as tautomers, stereoisomers such as R or S isomers with asymmetric carbon centers, geometric isomers (trans, cis), and optical isomers (enantiomers). All these isomers and mixtures thereof are also included within the scope of the present invention.

A composition for diagnosing neurodegenerative diseases according to another embodiment contains a compound represented by Formula 1, Formula 2, or Formula 3, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

At this time, the neurodegenerative disease is a degenerative brain disease, for example, Alzheimer's disease, Parkinson's disease (PD), Parkinson's disease dementia (PDD), Lewy body dementia (DLB), Lewy body variant of Alzheimer's disease (LBVAD), etc. It may be one or more diseases selected from the group consisting of systemic atrophy (MSA), Pick's disease, and tauopathy, and preferably, it may be Alzheimer's disease.

The compounds represented by Formula 1, Formula 2, or Formula 3 have excellent binding affinity to the Tau protein and are therefore useful in diagnosing neurodegenerative diseases, preferably Alzheimer's disease in which the Tau protein is expressed. It can be useful in the diagnosis of disease-related diseases.

A method of providing information for diagnosing a neurodegenerative disease according to another embodiment is

Administering a composition for diagnosing neurodegenerative diseases containing a compound represented by Formula 1, Formula 2, or Formula 3 as an active ingredient to a subject; It includes; measuring a signal for Tau protein.

The compounds represented by Formula 1, Formula 2, or Formula 3 have excellent binding affinity to the Tau protein and are therefore useful in diagnosing neurodegenerative diseases, preferably Alzheimer's disease in which the Tau protein is expressed. It can be useful in the diagnosis of disease-related diseases.

The signal measurement step is performed using one or more methods selected from the group consisting of single photon emission computed tomography (SPECT), positron emission tomography (PET), magnetic resonance imaging, and autoradiography. can be measured and imaged.

The signal is a signal in which the compound represented by Formula 1, Formula 2, or Formula 3 and tau protein or aggregates thereof are expressed, and through this, information regarding the diagnosis of neurodegenerative disease can be provided.

The neurodegenerative disease is a degenerative brain disease, for example, Alzheimer's disease, Parkinson's disease (PD), Parkinson's disease dementia (PDD), Lewy body dementia (DLB), Lewy body variant of Alzheimer's disease (LBVAD), and multiple system atrophy. It may be one or more diseases selected from the group consisting of (MSA), Pick's disease, and tauopathy, and preferably, it may be Alzheimer's disease.

The present invention will be described in more detail through examples below. The following examples are merely examples to aid understanding of the present invention, and the scope of the present invention is not limited thereto.

Example 1: [ ¹²⁵ I] KR201 pyrrolopyridine derivative compound production

As a precursor, 100 ug of the compound represented by the following formula 4 was dissolved in 250 uL of EtOH, 1.05 mCl of Na ¹²⁵ I was added, and then stirred. Next, 100uL of 1 N HCl and 3% H ₂ O ₂ were sequentially added and stirred at room temperature for 30 minutes. Next, the reaction was monitored by Radio TLC. At this time, Radio TLC conditions were that 0.1 M NaOMs was first spotted on the TLC, the reaction mixture was stirred, and then developed with 100% ethyl acetate. After the reaction was stirred, 100uL of 1 N NaOH was added to neutralize, filtered through a membrane filter, and purified by HPLC. At this time, the HPLC conditions were Xterra 10 um, 0 - 100% MeCN / 0.1% TFA in water for 25 min, 4 mL / min, 254 nm, Rt = 19.4 min. Next, after HPLC purification, diluted with 15 mL of water, passed through Light C-18 Sep-pak (used by activating with 10 mL of EtOH, 10 mL of water), captured, dried with N2 gas for 1 minute, and then washed with EtOH 1 Elution was performed in mL. [ ¹²⁵ I]KR201, a compound according to the following Chemical Formula 5, was prepared by drying EtOH with N2 gas at room temperature (RCY = 600 uCi (57.1%)).

[Formula 4]

[Formula 5]

Comparative Example 1: [ ¹²⁵ I]6-MK pyrrolopyridine derivative compound preparation

Compared to Example 1,

[ ¹²⁵ I]6-MK, a compound according to the following formula 7, was prepared in the same manner as in Example 1, except that the precursor formula 6 was used instead of the formula 4.

[Formula 6]

[Formula 7]

Example 2: [ ¹²³ I] KR201 pyrrolopyridine derivative compound production

Compared to Example 1,

The compound represented by Formula 4 was stirred after adding Na ¹²³ I 2.65 mCl instead of Na ¹²⁵ I 1.05 mCl; and

Finally, [ ¹²³ I]KR201, a compound according to the following formula 8, was prepared in the same manner as in Example 1, except that EtOH was dried with N2 gas at room temperature (RCY = 837 uCi (31.6%)). did.

[Formula 8]

Comparative Example 2: [ ¹²³ I]6-MK pyrrolopyridine derivative compound preparation

Compared to Example 2,

[ ¹²³ I]6-MK, a compound according to the following formula 9, was prepared in the same manner as in Example 2, except that formula 6 was used as a precursor instead of formula 4.

[Formula 9]

Example 3: [ ¹²⁴ I] KR201 pyrrolopyridine derivative compound production

Compared to Example 1, the compound represented by Formula 4 was stirred after adding 1.10 mCi of Na ¹²⁴ I instead of 1.05 mCi of Na ¹²⁵ I; Finally, [ ¹²⁴ I]KR201, a compound according to the following formula 10, was prepared in the same manner as in Example 1, except that EtOH was dried with N2 gas at room temperature (RCY = 320 uCi (29.0%)). .

[Formula 10]

Comparative Example 3: [ ¹²⁴ I]6-MK pyrrolopyridine derivative compound preparation

Compared to Example 3, [ ¹²⁴ I]6-MK, a compound according to the following Chemical Formula 11, was prepared in the same manner as Example 3, except that Chemical Formula 6 was used as the precursor instead of Chemical Formula 4.

[Formula 11]

Comparative Example 4: Control group of pyrrolopyridine derivative compounds [ ¹⁸ F]AV-1451

As a control for the new pyrrolopyridine derivative compound, AV-1451 (T807) from Lily company was used for comparative analysis.

[Formula 12]

Experimental Example 1-1: [ ¹²⁵ I]KR201 (Example 1) Confirmation of radioisotope labeling efficiency of pyrrolopyridine derivative compounds

[ ¹²⁵ I]KR201 (Example 1) A pyrrolopyridine derivative compound was prepared, the labeling efficiency was confirmed, and the results are shown in Figures 1A to 2B.

Specifically, Figure 1a is a graph showing the Radio TLC values measured 30 minutes after preparing the [ ¹²⁵ I]KR201 (Example 1) pyrrolopyridine derivative compound, and Figure 1b is a graph showing the Radio TLC values of Figure 1a and then HPLC purified. This is a graph showing the obtained values.

Referring to Figure 1a, as a result of checking TLC after 30 minutes, it was confirmed that most of the reaction had progressed (about 83% ROI). In addition, referring to Figure 1b, after confirming the TLC results, HPLC purification was performed and TLC was performed (the target compound with an Rf value of about 0.57) showed that a labeled compound with 100% purity was obtained after HPLC purification. .

Meanwhile, Figure 2a is a graph showing the RI peak after HPLC analysis of the [ ¹²⁵ I]KR201 (Example 1) pyrrolopyridine derivative compound, and Figure 2b is a graph showing the RI peak of the [ ¹²⁵ I]KR201 (Example 1) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis.

Referring to Figure 2a, it was confirmed that the RI peak was cleanly separated without any other impurities as a result of HPLC, and referring to Figure 2b, it was confirmed that there were no other overlapping peaks as a result of checking the UV peak, according to Example 1. [ ¹²⁵ I] It was confirmed that the KR201 pyrrolopyridine derivative compound has excellent specific activity and thus excellent labeling efficiency.

Experimental Example 1-2: [ ¹²⁵ I]6-MK (Comparative Example 1) Confirmation of radioisotope labeling efficiency of pyrrolopyridine derivative compounds

[ ¹²⁵ I]6-MK (Comparative Example 1) A pyrrolopyridine derivative compound was prepared, the labeling efficiency was confirmed, and the results are shown in Figures 3A to 4B.

Specifically, Figure 3a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²⁵ I]6-MK (Comparative Example 1) pyrrolopyridine derivative compound, and Figure 3b is a graph showing HPLC values after measuring the Radio TLC values of Figure 3a. This is a graph showing the values obtained through purification.

Referring to Figure 3a, as a result of checking TLC after 30 minutes, it was confirmed that most of the reaction had progressed (about 83% ROI). In addition, referring to Figure 3b, after confirming the TLC results, HPLC purification was performed and TLC was performed (the target compound with an Rf value of about 0.57) showed that a labeled compound with 100% purity was obtained after HPLC purification. .

Meanwhile, Figure 4a is a graph showing the RI peak after HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²⁵ I]6-MK (Comparative Example 1), and Figure 4b is a graph showing the RI peak of [ ¹²⁵ I]6-MK (Comparative Example 1) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis of a pyridine derivative compound.

Referring to Figure 4a, as a result of HPLC, it was confirmed that the RI peak was cleanly separated without any other impurities. Referring to Figure 4b, it was confirmed that there were no other overlapping peaks as a result of checking the UV peak, according to Comparative Example 1. It was confirmed that the [ ¹²⁵ I]6-MK pyrrolopyridine derivative compound has excellent specific activity and thus excellent labeling efficiency.

Experimental Example 1-3: [ ¹²³ I]KR201 (Example 2) Confirmation of radioisotope labeling efficiency of pyrrolopyridine derivative compounds

[ ¹²³ I]KR201 (Example 2) A pyrrolopyridine derivative compound was prepared, the labeling efficiency was confirmed, and the results are shown in Figures 5A to 6B.

Specifically, Figure 5a is a graph showing the Radio TLC values measured 30 minutes after preparing the [ ¹²³ I]KR201 (Example 2) pyrrolopyridine derivative compound, and Figure 5b is a graph showing the Radio TLC values of Figure 5a and then HPLC purified. This is a graph showing the obtained values.

Referring to Figure 5a, as a result of checking TLC after 30 minutes, it was confirmed that most of the reaction had progressed (about 49% ROI). In addition, referring to Figure 5b, after confirming the TLC results, HPLC purification was performed and TLC was performed (the target compound was found to have an Rf value of about 0.57), showing that a labeled compound with 94.08% purity was obtained after HPLC purification. .

Meanwhile, Figure 6a is a graph showing the RI peak after HPLC analysis of the [ ¹²³ I] KR201 (Example 2) pyrrolopyridine derivative compound, and Figure 6b is a graph showing the RI peak of the [ ¹²³ I] KR201 (Example 2) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis.

Referring to Figure 6a, it was confirmed that the RI peak was cleanly separated without other impurities as a result of HPLC, and referring to Figure 6b, it was confirmed that there were no other overlapping peaks as a result of checking the UV peak, according to Example 2. [ ¹²³ I] It was confirmed that the KR201 pyrrolopyridine derivative compound has excellent specific activity and thus excellent labeling efficiency.

Experimental Example 1-4: [ ¹²³ I] 6-MK (Comparative Example 2) Confirmation of radioisotope labeling efficiency of pyrrolopyridine derivative compounds

[ ¹²³ I]6-MK (Comparative Example 2) A pyrrolopyridine derivative compound was prepared, the labeling efficiency was confirmed, and the results are shown in Figures 7a to 8b.

Specifically, Figure 7a is a graph showing Radio TLC values measured 30 minutes after preparing the [ ¹²³ I] 6-MK (Comparative Example 2) pyrrolopyridine derivative compound, and Figure 7b is a graph showing HPLC values after measuring the Radio TLC values of Figure 7a. This is a graph showing the values obtained through purification.

Referring to Figure 7a, as a result of checking TLC after 30 minutes, it was confirmed that most of the reaction had progressed (about 78% ROI). In addition, referring to Figure 7b, after confirming the TLC results, HPLC purification was performed and TLC was performed (the target compound was found to have an Rf value of about 0.57), showing that a labeled compound with 100% purity was obtained after HPLC purification. .

Meanwhile, Figure 8a is a graph showing the RI peak after HPLC analysis of the pyrrolopyridine derivative compound of [ ¹²³ I]6-MK (Comparative Example 2), and Figure 8b is a graph showing the RI peak of [ ¹²³ I]6-MK (Comparative Example 2) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis of a pyridine derivative compound.

Referring to Figure 8a, as a result of HPLC, it was confirmed that some iodine remained in the RI peak, and it was confirmed that the peaks appearing after 20 minutes were impurities. However, referring to Figure 8b, as a result of checking the UV peak, other overlapping peaks were found. As it was confirmed that the [ ¹²³ I]6-MK pyrrolopyridine derivative compound according to Comparative Example 2 had a certain specific activity, it was confirmed that it had an average labeling efficiency.

Experimental Example 1-5: [ ¹²⁴ I]KR201 (Example 3) Confirmation of radioisotope labeling efficiency of pyrrolopyridine derivative compounds

[ ¹²⁴ I]KR201 (Example 3) A pyrrolopyridine derivative compound was prepared, the labeling efficiency was confirmed, and the results are shown in Figures 9a and 9b.

Specifically, Figure 9a is a graph showing the Radio TLC values measured 30 minutes after preparing the [ ¹²⁴ I]KR201 (Example 3) pyrrolopyridine derivative compound, and Figure 9b is a graph showing the Radio TLC values of Figure 9a and then HPLC purified. This is a graph showing the obtained values.

Referring to Figure 9a, as a result of checking TLC after 30 minutes, it was confirmed that most of the reaction had progressed (about 37% ROI). In addition, referring to Figure 9b, after confirming the TLC results, HPLC purification was performed and TLC was performed (the target compound was found to have an Rf value of about 0.57), and it can be seen that a labeled compound with a purity of 95% or more was obtained after HPLC purification. there was.

Meanwhile, Figure 9b is a graph showing the RI peak after HPLC analysis of the [ ¹²⁴ I]KR201 (Example 3) pyrrolopyridine derivative compound, and Figure 9c is a graph showing the RI peak of the [ ¹²⁴ I]KR201 (Example 3) pyrrolopyridine derivative compound. This is a graph showing the UV peak after HPLC analysis.

Referring to FIG. 9b, it was confirmed that the RI peak was cleanly separated without any other impurities as a result of HPLC, and referring to FIG. 9c, it was confirmed that there were no other overlapping peaks as a result of checking the UV peak, according to Example 3. [ ¹²⁴ I] It was confirmed that the KR201 pyrrolopyridine derivative compound has excellent specific activity and thus excellent labeling efficiency.

Experimental Example 2: Review of blood-brain barrier passage through Log P value

After preparing the pyrrolopyridine derivative compounds that underwent labeling according to Example 2 ([ ¹²³ I]KR201) and Comparative Example 2 ([ ¹²³ I]6-MK), 1 mCI of each was taken and mixed with PBS buffer (pH = 7.04). ) was dissolved in 1 mL of octanol and mixed well by vortexing. Next, 200 uL each was taken from the water layer and the octanol layer, the amount of activity was measured to obtain the log P value, and this was repeated a total of three times to obtain the average value (exp.) and the calculated and expected value ( calc.) is as shown in Table 3 below.

Comparative Example 2 ([ ¹²³ I] 6-MK) Example 2 ([ ¹²³ I] KR201) LogP (calc.) 3.87 3.87 LogP (exp.) 2.31 2.37

Usually, radiopharmaceuticals that target proteins in the brain must pass through the blood-brain barrier, and for this purpose, it is known that a log P value of about 2-3 is appropriate. Referring to Table 3 above, the log P value was measured. Results The expected value (calc.) calculated by calculating all the pyrrolopyridine derivative compounds that underwent labeling according to Example 2 ([ ¹²³ I]KR201) and Comparative Example 2 ([ ¹²³ I]6-MK) was 3.87, and the actual experiment It was confirmed that the average values were 2.31 and 2.37 for the 6-MK compound (Comparative Example 2) and the KR201 compound (Example 2), respectively.

Therefore, the average values for both the 6-MK compound (Comparative Example 2) and the KR201 compound (Example 2) are within the above range, confirming that both compounds can pass through the blood-brain barrier.

Experimental Example 3: Confirmation of target binding ability of Tau protein of Example 1 compound and Comparative Example 1 compound

The Tau protein is known as a microtubule-related protein in Alzheimer's dementia (AD) cases, and is a major component of the neurofibrillary tangle (NFT), performing axonal transport and stabilizing microtubules within nerve cells while repeating phosphorylation and dephosphorylation. plays a responsible role. Therefore, when the balance is broken by hyperphosphorylation of the tau protein, tau protein aggregation and NFTs are generated and accumulated in the brain, which can induce neuronal cell death. Therefore, in order to confirm whether the pyrrolopyridine derivative compound has the target binding ability of Tau protein according to Example 1 ([ ¹²⁵ I] KR201) and Comparative Example 1 ([ ¹²⁵ I] 6-MK), Example 1 ([ ¹²⁵ After preparing a pyrrolopyridine derivative compound according to I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK), autoradiography and immunohistochemistry were performed.

The autoradiography method was performed using 100% iced cold acetone, fixed at room temperature for 2 minutes, and washed three times for 5 minutes with PBS. Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ According to [ ¹²⁵ I]6-MK), 372 μCi and 430 μCi of pyrrolopyridine derivative compounds were treated, respectively. Next, for the blocking assay, each slide was treated with cold compounds (cold ^-125 I-6-MK, cold- ¹²⁵ I-KR201) at a concentration of 50 μM for 30 minutes, and then reacted at room temperature for 60 minutes. Afterwards, the slides were washed in the following order: PBS, 70% EtOH, 30% EtOH, and DW, and dried at room temperature. Next, after exposing for 24 hours using phosphor imaging plates, images were confirmed using a Typhoon FLA 9500 (GE Healthcare) imaging system, and the results are shown in Figure 10 below.

Specifically, Figure 10 shows the target-binding ability of Tau protein through autoradiography of the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK). This is an image showing the results of confirmation.

Referring to Figure 10, [ ¹²⁵ I]6-MK (Comparative Example 1), [ ¹²⁵ I]KR201 for Tau protein in human brain tissue (hippocampus, frontal cortex) containing NFT. (Example 1) Strong binding to the pyrrolopyridine derivative compound was confirmed.

In addition, the immunohistochemistry was performed on the tissue section slides performed in Example 1 ([ ¹²⁵ I] KR201) and Comparative Example 1 ([ ¹²⁵ I] 6-MK) by deparaffinizing and dehydrating the slides with distilled water. Washed. Next, to eliminate the activity of endogenous peroxidase, peroxide blocking (DAKO) was applied for 10 minutes at room temperature. Next, after washing twice with PBS, the antibody PHF-Tau (invitrogen, Cat. ab2539, 1:1000) was incubated at 4°C overnight, washed with Wash buffer (DAKO), and reacted with Envision+ Rabbit for 30 minutes. Next, it was washed with wash buffer (DAKO), and color was developed with DAB (3,3-diaminibenzidine tetrahydrochloride) for 3 minutes. Next, it was neutralized with distilled water, counterstained with Mayer hematoxylin, washed with tap water, and encapsulated through a dehydration and transparency process to measure the radioactivity concentration. The results are shown in Figure 11, Table 4, and Table 5. indicated.

Specifically, Figure 11 shows the target of Tau protein through immunohistochemistry of the pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK). This is a graph comparing the radioactivity concentration shown as a result of confirming the binding ability.

PSL/mm2 (Mean±SD) KR201-H 1480.51±475.24 6-MK-H (Ref.) 642.367±48.19 KR201-F (New comp.) 2460.55±442.71 6-MK-F (Ref.) 1531.54±230.07

New comp. to Ref.PSL/mm ² ratio Hippocampus 2.34±0.91 Frontal cortex 1.65±0.54

Referring to FIG. 11, Table 4, and Table 5, as a result of quantitative analysis in the region of interest (ROI), the radioactivity concentration according to Comparative Example 1 ([ ¹²⁵ I] 6-MK) was found in both the hippocampus and the frontal lobe. The pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I] KR201) showed a value about two times higher than that of the rolopyridine derivative compound.

That is, summarizing the above, the Tau protein binding site of [ ¹²⁵ I]6-MK (Comparative Example 1) and [ ¹²⁵ I]KR201 (Example 1) pyrrolopyridine derivative compounds through magnetoradiography; and immune tissue Through chemical analysis, it was confirmed that the location where Tau was overexpressed was consistent, and in particular, in the pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I]KR201), Comparative Example 1 ([ ¹²⁵ I]6-MK) It was confirmed that the level was approximately twice higher than that of the pyrrolopyridine derivative compound according to .

Experimental Example 4: Confirmation of absorption and distribution of compounds of Example 1 and Comparative Example 1

Absorption and distribution in each organ when the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK) were administered to animals (mouse) at a certain dosage The confirmation results are shown in Figures 12 and 13 and Tables 6 and 7.

Specifically, the absorption and distribution of each compound were confirmed as follows.

In normal male mice (8 weeks old, 20-25g), the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK) were administered through the tail vein. Injected. At this time, when the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK) were administered to animals at a dose of 0.1 mCi/mous, during the experiment period There were no unusual symptoms or deaths in the animals, and no abnormal symptoms were observed in the autopsy performed after completion of the test.

Animals were sacrificed at 2, 10, 30, 90, and 120 minutes after injection and blood, muscle, heart, lung, liver, spleen, stomach, intestine, kidney, bone, brain (cerebrum, striatum and others) were sacrificed. Brain tissue area) was extracted, weighed, placed in a plastic vial, measured for radioactivity, and %ID/g was calculated in the following manner. Radioactivity was measured using a gamma counter (WIZARD 1480, Perkin-Elmer, USA).

1. Prepare a syringe containing 0.1 mCi/0.1 mL [ ¹²⁵ I]6-MK and [ ¹²⁵ I]KR201.

2. Dilute the [ ¹²⁵ I]6-MK and [ ¹²⁵ I]KR201 to a total volume of 0.1 L using a 0.2 L volumetric flask.

3. Take 0.1 mL from the above solution using a syringe and measure the weight: ①

4. Prepare the standard solution in three test tubes and measure the weight of the remaining syringe: ②

5. Count the solution in three test tubes using a gamma counter: ③

6. Before injecting [ ¹²⁵ I]6-MK, [ ¹²⁵ I]KR201 into the animal, measure the weight of the syringe containing [ ¹²⁵ I]6-MK, [ ¹²⁵ I]KR201: ④

7. Measure the weight of the remaining syringe after injecting the animal: ⑤

8. Remove tissue and measure weight: ⑥

9. Count the radioactivity of the tissue: ⑦

10. How to get %ID/g:

Specifically, Figure 12 is a graph showing the results of radioactivity distribution in each organ over time after administering the pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I] KR201) to an animal. Additionally, the results are shown in Table 6 below.

%ID/g 2m 10m 30m 90m 120 m Blood 4.51±1.95 3.08±0.51 3.35±0.88 3.22±0.34 3.54±0.43 Muscle 1.34±0.76 1.50±0.17 0.97±0.15 0.84±0.29 0.82±0.23 Fat 0.72±0.08 2.78±0.95 2.61±1.42 1.33±0.22 0.99±0.21 Heart 9.83±3.49 2.58±0.20 1.56±0.26 1.29±0.22 1.20±0.19 Lung 77.88±2.52 6.76±0.89 4.18±1.18 3.18±0.50 3.85±0.57 Liver 16.08±4.44 34.86±10.17 30.67±4.88 32.34±2.97 21.65±2.22 Spleen 7.31±1.74 3.84±0.75 2.06±0.27 1.67±0.19 1.66±0.15 Stomach 4.37±2.00 5.63±0.55 16.32±4.36 23.87±3.05 24.80±3.80 Intestine 6.23±1.26 7.19±1.24 8.12±1.67 13.97±2.63 18.06±2.75 Kidney 37.29±7.98 10.87±2.17 5.37±1.91 3.35±0.74 2.71±0.36 Bone 2.27±0.36 1.88±0.55 1.58±0.32 1.36±0.24 1.44±0.25 Cerebellum 10.46±0.93 3.96±0.52 0.86±0.40 0.23±0.03 0.16±0.03 Striatum 11.91±1.08 4.56±0.60 0.99±0.44 0.25±0.03 0.19±0.04 Rest of brain 12.80±1.13 4.67±0.52 1.03±0.46 0.31±0.04 0.28±0.04

In addition, Figure 13 is a graph showing the results of radioactivity distribution in each organ over time after administering the pyrrolopyridine derivative compound according to Comparative Example 1 ([ ¹²⁵ I]6-MK) to an animal. Additionally, the results are shown in Table 7 below.

%ID/g 2m 10m 30m 90 m 120 m Blood 3.38±2.25 3.71±0.62 2.55±0.27 2.02±0.30 1.81±0.49 Muscle 0.71±0.46 1.02±0.18 0.63±0.15 0.64±0.05 0.42±0.21 Fat 0.57±0.46 0.86±0.25 1.58±0.27 1.25±0.22 0.96±0.54 Heart 8.06±8.05 2.35±0.39 1.07±0.13 0.75±0.08 0.64±0.21 Lung 13.85±9.47 8.80±4.06 3.66±0.80 2.60±0.55 1.91±0.42 Liver 14.60±9.75 22.64±3.44 10.09±0.63 5.25±0.86 3.70±1.34 Spleen 3.73±2.52 2.50±0.07 1.56±0.19 1.12±0.15 0.89±0.28 Stomach 2.96±2.15 8.49±3.29 24.51±12.14 15.18±7.01 13.22±5.89 Intestine 3.45±2.30 5.43±0.81 13.52±2.69 18.88±3.21 18.92±3.06 Kidney 21.27±15.04 14.93±1.92 8.73±1.07 3.68±1.49 2.21±1.30 Bone 1.48±0.84 1.02±0.08 1.09±0.27 0.77±0.14 0.63±0.32 Cerebellum 5.03±3.51 2.37±0.38 0.47±0.18 0.14±0.04 0.10±0.06 Striatum 5.29±3.69 2.63±0.33 0.51±0.22 0.19±0.13 0.11±0.06 Rest of brain 5.89±4.12 2.80±0.45 0.58±0.20 0.18±0.04 0.15±0.09

Referring to Figures 12 and 13 and Tables 6 and 7, most of the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I] KR201) and Comparative Example 1 ([ ¹²⁵ I] 6-MK) after the initial injection. Each organ showed high radioactivity at 2 minutes. However, at this time, the distribution of the pyrrolopyridine derivative compounds according to Example 1 ([ ¹²⁵ I]KR201) and Comparative Example 1 ([ ¹²⁵ I]6-MK) is simple diffusion through the bloodstream rather than being selectively transferred to the relevant organ. Since it is caused by , it can be seen as distributed in each organ. After 2 minutes, it tended to be discharged quickly from most organs, and after 30 minutes after injection, blood (6-MK: 2.55 ± 0.27, KR201: 3.35 ± 0.88 %ID/g) and heart (6-MK: 1.07 ± 0.13, KR201: 1.56 ± 0.26 %ID/g) showed only very low levels of radiopharmaceutical residue. In the first 2 minutes after the injection containing Comparative Example 1 ([ ¹²⁵ I]6-MK), it was rapidly absorbed and distributed to each region of the brain (cerebrum, striatum and rest of the brain), and about 5 to 6 doses were administered in each region. While it showed an intake of %ID/g, it showed an intake of about 10-13%ID/g after injection containing Example 1 ([ ¹²⁵ I]KR201), which was more than twice as high as that of the 6-MK compound. check. Meanwhile, intestine, the intake of about 18 %ID/g was maintained for both compounds from the initial injection to 120 minutes, and the liver showed 22.64 ± 3.44 %ID/g (6-MK) from the initial injection to 10 minutes after injection. Compound), shows an intake of 34.86 ± 10.17 %ID/g (KR201 compound), and over time, 3.70 ± 1.34 %ID/g (6-MK compound), 21.65 ± 2.22 %ID/g (KR201 compound) in 120 minutes. ) I was able to confirm the results being emitted. In Kidney, both compounds were rapidly excreted from 10 minutes and showed a tendency to rapidly decrease after 30 minutes, with values below 1% ID/g. In blood, [ ¹²⁵ I]6-MK and [ ¹²⁵ I]KR201 showed concentrations of 3.38 ± 2.25, 4.51 ± 1.95 %ID/g 2 minutes after injection, and 1.81 ± 0.49, 3.54 ± 0.43 %ID/g at 120 minutes. /g intake was shown. As a result, in the case of the pyrrolopyridine derivative compound according to Example 1 ([ ¹²⁵ I] KR201), the pyrrolopyridine derivative compound according to Comparative Example 1 ([ ¹²⁵ I] 6-MK) In comparison, it was confirmed that the intake was more than twice as high for each part of the brain (cerebrum, striatum, rest of the brain) compared to other organs.

That is, the pyrrolopyridine derivative compounds ([ ¹²⁵ I]KR201, [ ¹²³ I]KR201) according to one embodiment not only showed high labeling efficiency as a result of a labeling experiment using a radioisotope, but were also able to penetrate the blood-brain barrier. The LogP value, which is an indicator, was also confirmed to be a good value of 2-3.

In addition, the pyrrolopyridine derivative compounds ([ ¹²⁵ I]KR201, [ ¹²³ I]KR201) according to one embodiment are the pyrrolopyridine derivative compounds ([ ¹²⁵ I]6-MK, [ ¹²³ I]6-MK), which are comparative examples. Compared to this, it showed high uptake in each area of the brain (cerebrum, striatum, rest of brain) and high Tau binding ability in postmortem brain tissue slides of Alzheimer's patients. Based on these results, a composition containing this for diagnosing neurodegenerative diseases was used. Thus, it will be possible to easily provide information for diagnosing neurodegenerative diseases.

Experimental Example 5: PET image evaluation of compounds of Example 1 and Comparative Example 2 in normal mice

Pyrrolopyridine derivative compounds according to Example 3 ([ ¹²⁴ I]KR201) and Comparative Examples 3 ([ ¹²⁴ I]6-MK) and 4 ([ ¹⁸ F]AV-1451) and Comparative Example 2 samples in clinical use When administered to the tail vein of an animal (mouse) at a certain dosage, the results of disappearance after ingestion into the brain after positron emission tomography (PET) are shown in Figures 14, 15, and 16 and Table 8 (KR201) and Table 9 (6-MK). ) and Table 10 (AV-1451).

Specifically, biological imaging of each compound was confirmed as follows. In each of Example 3 ([ ¹²⁴ I]KR201) and Comparative Examples 3 ([ ¹²⁴ I]6-MK) and 4 ([ ¹⁸ F]AV-1451) in normal male mice (8 weeks old, 20-25 g), The following pyrrolopyridine derivative compound was injected through the tail vein. At this time, the compounds according to Example 1 ([ ¹²⁴ I]KR201) and Comparative Examples 3 ([ ¹²⁴ I]6-MK) and 4 ([ ¹⁸ F]AV-1451) were administered at doses of 150 uCi, 150 uCi, 1000. When uCi was administered to animals, no unusual symptoms or deaths occurred during the experiment, and no abnormal symptoms were observed even after imaging.

The injection was performed using an injection pump at the same time as the start of imaging, and was injected at the same rate for a total of 1 minute. Each compound was subjected to dynamic PET/CT imaging for 90 minutes in Example 3 ([ ¹²⁴ I]KR201) or Comparative Examples 3 ([ ¹²⁴ I]6-MK) and 4 ([ ¹⁸ F]AV-1451), and Examples 3 ([ ¹²⁴ I]KR201) underwent static PET/CT scanning for 30 minutes after 24 hours. Each image was reconstructed at 2-minute intervals for the first 10 minutes (2, 4, 6, 8, 10 minutes), and at 10-minute intervals after 10 minutes.

The standardized uptake value (SUV) of each image was calculated using the amount of drug injected into the mouse and its body weight. To confirm the target intake amount to the brain of normal mice of Example 3 ([ ¹²⁴ I]KR201) and Comparative Examples 3 ([ ¹²⁴ I]6-MK) and 4 ([ ¹⁸ F]AV-1451), each time A VOI (Volume of Interest) was manually drawn on the entire brain area in the image, and the corresponding value (SUV) was displayed as a graph (Time Activity Curves).

time (hour) SUV (mean±SD)
0 0 0.03 0.88±0.16 0.07 1.61±0.20 0.10 1.42±0.21 0.13 1.23±0.23 0.17 1.09±0.18 0.33 0.73±0.03 0.50 0.46±0.01 0.67 0.35±0.03 0.83 0.30±0.04 1.00 0.28±0.04 1.17 0.26±0.03 1.33 0.24±0.03 1.50 0.24±0.06 24.00 0.17±0.11

time (hour) SUV (mean±SD)
0 0 0.03 0.62±0.11 0.07 0.82±0.14 0.10 0.63±0.11 0.13 0.51±0.08 0.17 0.44±0.07 0.33 0.30±0.04 0.50 0.22±0.02 0.67 0.19±0.02 0.83 0.17±0.01 1.00 0.16±0.01 1.17 0.16±0.01 1.33 0.15±0.01 1.50 0.14±0.01

time (hour) SUV (mean±SD)
0 0 0.03 1.38±0.26 0.07 2.46±0.14 0.10 2.16±0.16 0.13 1.79±0.14 0.17 1.50±0.17 0.33 0.97±0.13 0.50 0.55±0.07 0.67 0.32±0.10 0.83 0.25±0.07 1.00 0.22±0.06 1.17 0.22±0.06 1.33 0.17±0.07 1.50 0.17±0.07

Experimental Example 6: PET image evaluation of compounds of Example 1 and Comparative Example 2 in transgenic mice

Pyrrolopyridine derivative compounds according to Example 3 ([ ¹²⁴ I]KR201) and Comparative Example 4 ([ ¹⁸ F]AV-1451) and Comparative Example 2 samples in clinical use were administered to transgenic animals (mouse) at a certain dose. When administered through the tail vein, the results of disappearance after ingestion into the brain after positron emission tomography (PET) are shown in Figures 17 and 18 and Tables 11 (KR201) and 12 (AV-1451).

Specifically, biological imaging of each compound was confirmed as follows. Each example 3 ([ ¹²⁴ I]KR201) in a transgenic mouse overexpressing human tau protein (Tg(Prnp-MAPT*P301L)JNPL3Hlmc, 23 weeks old, 54 g) and a transgenic mouse overexpressing human tau protein (15 weeks old) , 46 g, same individual), the pyrrolopyridine derivative compound according to Comparative Example 4 ([ ¹⁸ F]AV-1451) was injected through the tail vein. At this time, when the compounds according to Example 1 ([ ¹²⁴ I]KR201) and Comparative Example 4 ([ ¹⁸ F]AV-1451) were administered to animals at doses of 150 uCi and 1000 uCi, respectively, the animals' No unusual symptoms or deaths occurred, and no abnormal symptoms were observed even after imaging.

The injection was performed using an injection pump at the same time as the start of imaging, and was injected at the same rate for a total of 1 minute. Each compound was subjected to dynamic PET/CT imaging for 90 minutes in Example 3 ([ ¹²⁴ I]KR201) or Comparative Example 4 ([ ¹⁸ F]AV-1451). Each image was reconstructed at 2-minute intervals for the first 10 minutes (2, 4, 6, 8, 10 minutes), and at 10-minute intervals after 10 minutes.

The standardized uptake value (SUV) of each image was calculated using the amount of drug injected into the mouse and its body weight. In order to confirm the target intake amount to the brain of normal mice in Example 3 ([ ¹²⁴ I]KR201) and Comparative Example 4 ([ ¹⁸ F]AV-1451), whole brain VOI was performed on the entire brain area of the images at each time. (Volume of Interest) was drawn manually, and the corresponding value (SUV) was displayed as a graph (Time Activity Curves).

time (hours) SUV (mean) 0 0 0.03 1.45 0.07 2.31 0.10 1.84 0.13 1.52 0.17 1.31 0.33 0.80 0.50 0.49 0.67 0.37 0.83 0.30 1.00 0.28 1.17 0.26 1.33 0.25 1.50 0.24

time (hours) SUV (mean) 0 0 0.03 1.65 0.07 2.83 0.10 2.41 0.13 2.02 0.17 1.68 0.33 1.11 0.50 0.65 0.67 0.49 0.83 0.44 1.00 0.41 1.17 0.39 1.33 0.37 1.50 0.37

Claims (13)

A compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
[Formula 1]

X is CH, O, or S,
R ¹ and R ² are each independently hydrogen, unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, or cyano group. (-CN);
When R ¹ and R ² are connected to form a ring, they are an unsubstituted or substituted homoaryl group of 5 or 10 atoms, or an unsubstituted or substituted 5 or 6 atom optionally containing N, S and/or O. It is a heteroaryl group of atoms;
R ³ is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C _1-6 straight or branched alkyl, unsubstituted or substituted C _2-6 straight or branched alkenyl, unsubstituted or substituted C _1-6 linear or branched alkoxy, unsubstituted or substituted C _6-10 aryl group, or unsubstituted or substituted 5- or 6-membered heteroaryl group optionally containing N, S and/or O. ego; and
The substituted substituent is substituted with one or more selected from the group consisting of halogen, hydroxy, C _1-5 straight or branched alkyl, unsubstituted or C 1-5 straight or branched alkoxy substituted with one or more halogens. . According to paragraph 1,
wherein X is CH;
R ¹ and R ² are connected to form an unsubstituted or substituted 5- or 10-atom homoaryl group;
R ³ is hydrogen, halogen, hydroxy group (OH), unsubstituted or substituted C _1-6 straight or branched alkyl, or unsubstituted or substituted C _1-6 straight or branched alkoxy; and
The substituted substituents are at least one selected from the group consisting of halogen, hydroxy group (OH), C _1-5 straight or branched alkyl, unsubstituted or C _1-5 straight or branched alkoxy substituted with one or more halogens. A compound substituted with, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. According to paragraph 1,
A compound represented by the following formula (2), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
[Formula 2]

wherein R ^a1 to R ^a4 are each independently hydrogen or a halogen group;
R ^b is hydrogen, halogen, hydroxy group (OH), or unsubstituted or substituted C _1-6 straight or branched alkoxy; and
The substituted substituents are at least one selected from the group consisting of halogen, hydroxy group (OH), C _1-5 straight or branched alkyl, unsubstituted or C _1-5 straight or branched alkoxy substituted with one or more halogens. A compound substituted with or a pharmaceutically acceptable salt thereof. According to paragraph 3,
Wherein R ^a1 , R ^a2 , and R ^a4 are hydrogen, R ^a2 is a halogen group; and
wherein R ^b is hydrogen, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. According to paragraph 4,
A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the halogen group is one selected from the group consisting of F, Cl, Br, and I. According to clause 5,
The halogen group is a compound selected from the group consisting of isotopically labeled ¹⁸ F, ⁸² Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I, and ¹³¹ I, a stereoisomer thereof, or a pharmaceutical product thereof. Salts that are generally acceptable. According to paragraph 1,
A compound represented by the following formula (3), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.
[Formula 3]

Said R is ¹²³ I, ¹²⁴ I or ¹²⁵ I. A composition for diagnosing neurodegenerative diseases, containing the compound according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. According to clause 8,
A composition for diagnosing neurodegenerative diseases, wherein the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof binds to the Tau protein. According to clause 8,
A composition for diagnosing a neurodegenerative disease, wherein the neurodegenerative disease is a degenerative brain disease. According to clause 8,
The neurodegenerative diseases include Alzheimer's disease, Parkinson's disease (PD), Parkinson's disease dementia (PDD), Lewy body dementia (DLB), Lewy body variant of Alzheimer's disease (LBVAD), multiple system atrophy (MSA), Pick disease, and tau. A composition for diagnosing a neurodegenerative disease, which is one or more diseases selected from the group consisting of diseases. Administering the composition for diagnosing neurodegenerative diseases of claim 8 to a subject; and
A method of providing information for diagnosing neurodegenerative diseases, comprising: measuring a signal for Tau protein. According to clause 12,
The signal measurement step is performed using one or more methods selected from the group consisting of single photon emission computed tomography (SPECT), positron emission tomography (PET), magnetic resonance imaging, and autoradiography. A method of providing information for diagnosing neurodegenerative diseases, which involves measuring and imaging.