Compound for detecting beta-amyloid oligomer and method for diagnosing neurodegenerative disease by using compound

Abstract

The present invention relates to a novel compound for detecting beta-amyloid protein and a method for diagnosing neurodegenerative diseases using the same, in which a compound represented by chemical formula 1 or a salt thereof according to the present invention can specifically bind to beta-amyloid protein. Since the compound or the salt thereof has high selectivity, beta-amyloid protein in plasma can be detected with high sensitivity even if detection is blocked due to the presence of other proteins in plasma, and thus the compound or the salt thereof can be used as a therapeutic agent for diagnosing neurodegenerative diseases or screening for neurodegenerative diseases.

Description

Technical Field

[0001] This invention relates to a novel compound for detecting β-amyloid oligomers and a method for diagnosing neurodegenerative diseases using the compound. Background Technology

[0002] Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by the deposition of amyloid beta (Aβ) peptides in the brain. Aβ peptides are produced when amyloid precursor proteins are cleaved by enzymes, and these peptides aggregate in AD patients to form oligomers and plaques. Although there is no clear correlation between Aβ plaque burden and cognitive impairment, Aβ oligomers are known to be involved in the pathogenesis of AD symptoms. Furthermore, in the brains of AD patients, Aβ oligomers contribute to neurotoxicity, neuronal death, and synaptic dysfunction. Therefore, detecting Aβ oligomers is crucial for developing AD therapeutics and diagnostic methods.

[0003] Given that the main Aβ species in plasma are soluble Aβ oligomers, detecting plasma Aβ can be used to monitor changes in Aβ oligomer levels and predict the onset of Alzheimer's disease (AD). Furthermore, blood Aβ testing is a non-invasive and cost-effective method for detecting Aβ oligomers in clinical trials, which could benefit AD patients. However, accurately measuring Aβ oligomer levels in bloodstream remains a challenge in terms of accuracy and sensitivity. The controversy surrounding accurate plasma Aβ measurement is well-known due to the heterogeneity and polymorphic properties of Aβ oligomers. Specifically, Aβ oligomers are unstable intermediates with various shapes, structures, and compositions, unlike the β-sheet-rich structure of Aβ plaques. The transient and polymorphic nature of Aβ oligomers leads to the masking of Aβ oligomer-targeting compounds and creates steric hindrance, resulting in low diagnostic accuracy and sensitivity. Therefore, there is an urgent need to develop new imaging agents to selectively target Aβ oligomers.

[0004] Therefore, the inventors of this invention provide a compound that not only significantly improves detection efficiency by interacting with Aβ aggregates, but also maintains and does not reduce the detection efficiency for Aβ-modified complex forms. Summary of the Invention

[0005] Technical issues

[0006] On the one hand, it provides compounds represented by the following chemical formula 1 or their salts:

[0007] [Chemical Formula 1]

[0008] .

[0009] On the other hand, a method for preparing the compound represented by the chemical formula 1 is provided.

[0010] On the other hand, a composition for detecting β-amyloid (Aβ) is provided, comprising a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0011] On the other hand, a method for detecting β-amyloid protein (Aβ) is provided, comprising the steps of contacting an isolated biological sample with a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0012] Another aspect provides a composition for diagnosing neurodegenerative diseases, comprising a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0013] Another approach provides a method for providing information on the diagnosis of neurodegenerative diseases, comprising the step of contacting an isolated biological sample with a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0014] Another approach provides a method for diagnosing neurodegenerative diseases, comprising the steps of administering a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof to a biological sample.

[0015] Technical solution

[0016] On the one hand, it provides compounds represented by the following chemical formula 1 or their salts:

[0017] [Chemical Formula 1]

[0018]

[0019] In the chemical formula 1,

[0020] The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0021] The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2.

[0022] [Chemical Formula 2]

[0023]

[0024] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus,

[0025] The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0026] The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0027] The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

[0028] In one specific example, the R 1a and R 1b Each can be independently hydrogen, deuterium, or halogen;

[0029] The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-5 Alkyl, substituted or unsubstituted C 2-5 alkenyl, substituted or unsubstituted C 2-5 Alkyne, substituted or unsubstituted C 1-5 Alkoxy, substituted or unsubstituted 6-membered aryl, or substituted or unsubstituted 6-membered heteroaryl, or said R 1c and R 1d They connect to each other to form the compound represented by chemical formula 2.

[0030] The X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus,

[0031] The R 4 It can be hydrogen, deuterium, or halogen;

[0032] The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, substituted or unsubstituted C. 1-5 Alkyl, substituted or unsubstituted C 2-5 alkenyl, substituted or unsubstituted C 2-5 Alynyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0033] The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-5 Alkyl), substituted or unsubstituted C 1-5 Alkyl or substituted or unsubstituted 6- to 10-membered aryl groups.

[0034] In another specific example, the R 1a and R 1b Each can be either hydrogen or halogen;

[0035] The R 1c and R 1dEach is independently hydrogen, halogen, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted C 1-5 Alkoxy or substituted or unsubstituted 6-membered aryl group, or the R 1c and R 1d They connect to each other to form the compound represented by chemical formula 2.

[0036] The X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ),

[0037] The R 4 It is hydrogen;

[0038] The R 2a and R 2b Each is independently hydrogen, halogen, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 6-membered heteroaryl;

[0039] The R 3a and R 3b They can each independently be hydrogen, -CN, -C(O)NH2, -C(O)O(C 1-5 Alkyl), substituted or unsubstituted C 1-5 Alkyl or substituted or unsubstituted 6-membered aryl groups.

[0040] The term "alkyl" refers to a functional group in which a hydrogen atom has been removed from a carbon atom in a saturated hydrocarbon group, where the carbon atoms are linked by a single bond. The alkyl group can contain 1 to 20 carbon atoms (C1 to C2). 1-20 ), 1 to 10 carbon atoms (C 1-10 ) or 1 to 5 carbon atoms (C 1-5 ( ) is a straight-chain or chain hydrocarbon. For example, it can be methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t-butyl, etc.

[0041] The term "Cm" - n (where m and n are each an integer greater than or equal to 1) means having m to n carbon atoms.

[0042] The term "alkenyl" refers to a functional group consisting of an unsaturated hydrocarbon group having a hydrogen atom removed from one carbon atom, the unsaturated hydrocarbon group being connected by one or more double bonds between carbon atoms. The alkenyl group can contain 2 to 20 carbon atoms (C2+10+2 ... 2-20 ), 2 to 10 carbon atoms (C 2-10 ) or 2 to 5 carbon atoms (C 2-5 Straight-chain or chain hydrocarbons. For example, they can be vinyl, propenyl, butenyl, etc.

[0043] The term "alkynyl" refers to a functional group consisting of an unsaturated hydrocarbon group having a hydrogen atom removed from one carbon atom, the unsaturated hydrocarbon group being connected by one or more triple bonds between carbon atoms. The alkynyl group can contain 2 to 20 carbon atoms (C2+12+20 ... 2-20 ), 2 to 10 carbon atoms (C 2-10 ) or 2 to 5 carbon atoms (C 2-5 ( ) are straight-chain or chain hydrocarbons. For example, they can be ethynyl, propynyl, butinyl, etc.

[0044] The term "alkoxy group" refers to an alkyl group (-O-alkyl) bonded to oxygen. The alkoxy group may contain 2 to 20 carbon atoms (C60-C60). 2-20 ), 2 to 10 carbon atoms (C 2-10 ) or 2 to 5 carbon atoms (C 2-5 For example, it can be methoxy, ethoxy, propoxy, butoxy, etc.

[0045] The term "aryl" refers to a functional group formed by removing a hydrogen atom from a carbon atom in a parent aromatic hydrocarbon. Depending on the number of atoms forming the ring, the aryl group can be 5- to 18-membered, 5- to 14-membered, 5- to 10-membered, or 6- to 10-membered. For example, it can be phenyl, biphenyl, naphthalenyl, fluorenyl, indenyl, indanyl, etc.

[0046] The term "heteroaryl" refers to an aryl group in which one or more carbon atoms constituting an aromatic ring are replaced by atoms selected from the group consisting of nitrogen, oxygen, and sulfur. Depending on the number of atoms constituting the ring, the heteroaryl group can be 5-membered to 18-membered, 5-membered to 14-membered, or 5-membered to 10-membered. For example, it can be thiopheneyl, pyrrolyl, pyrazolyl, pyridinyl, imidazolyl, triazolyl, oxazolyl, etc.

[0047] The alkyl, alkenyl, alkynyl, alkoxy, aryl, or heteroaryl groups may or may not be substituted. The term "substituted" means that one or more hydrogen atoms bonded to a carbon atom are replaced by another atom or functional group, and "unsubstituted" means that none of the hydrogen atoms bonded to a carbon atom are replaced by another atom or functional group. Examples of other substituted atoms may be fluorine, chlorine, bromine, or iodine; examples of substituted functional groups may be hydroxyl, thiol, nitro, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy, substituted or unsubstituted alkylthio, substituted or unsubstituted aryloxy, substituted or unsubstituted arylthio, substituted or unsubstituted heteroaryloxy, or substituted or unsubstituted heteroarylthio, etc.

[0048] Examples of substituted alkyl groups include trifluoromethyl (-CF3) or (2-(2-(2-fluoroethoxy)ethoxy)ethoxy)methyl (-CH2OC2H4OC2H4OC2H5F), etc.

[0049] Examples of aryl group substitution include methoxy-phenyl, fluoro-phenyl, chloro-phenyl, or tolyl.

[0050] In one specific example, the compound represented by chemical formula 1 may be a compound represented by one of the chemical formulas selected from the group consisting of chemical formulas 3 to 43:

[0051] [Chemical Formula 3]

[0052] ,

[0053] [Chemical Formula 4]

[0054] ,

[0055] [Chemical Formula 5]

[0056] ,

[0057] [Chemical Formula 6]

[0058] ,

[0059] [Chemical Formula 7]

[0060] ,

[0061] [Chemical Formula 8]

[0062] ,

[0063] [Chemical Formula 9]

[0064] ,

[0065] [Chemical Formula 10]

[0066] ,

[0067] [Chemical Formula 11]

[0068] ,

[0069] [Chemical Formula 12]

[0070] ,

[0071] [Chemical Formula 13]

[0072] ,

[0073] [Chemical Formula 14]

[0074] ,

[0075] [Chemical Formula 15]

[0076] ,

[0077] [Chemical Formula 16]

[0078] ,

[0079] [Chemical Formula 17]

[0080] ,

[0081] [Chemical Formula 18]

[0082] ,

[0083] [Chemical Formula 19]

[0084] ,

[0085] [Chemical Formula 20]

[0086] ,

[0087] [Chemical Formula 21]

[0088] ,

[0089] [Chemical Formula 22]

[0090] ,

[0091] [Chemical Formula 23]

[0092] ,

[0093] [Chemical Formula 24]

[0094] ,

[0095] [Chemical Formula 25]

[0096] ,

[0097] [Chemical Formula 26]

[0098] ,

[0099] [Chemical Formula 27]

[0100] ,

[0101] [Chemical Formula 28]

[0102] ,

[0103] [Chemical Formula 29]

[0104] ,

[0105] [Chemical Formula 30]

[0106] ,

[0107] [Chemical Formula 31]

[0108] ,

[0109] [Chemical Formula 32]

[0110] ,

[0111] [Chemical Formula 33]

[0112] ,

[0113] [Chemical Formula 34]

[0114] ,

[0115] [Chemical Formula 35]

[0116] ,

[0117] [Chemical Formula 36]

[0118] ,

[0119] [Chemical Formula 37]

[0120] ,

[0121] [Chemical Formula 38]

[0122] ,

[0123] [Chemical Formula 39]

[0124] ,

[0125] [Chemical Formula 40]

[0126] ,

[0127] [Chemical Formula 41]

[0128] ,

[0129] [Chemical Formula 42]

[0130] as well as

[0131] [Chemical Formula 43]

[0132] .

[0133] The salt can be an acid addition salt formed from a free acid. When the compound is used to diagnose a disease, it can be in the form of a pharmaceutically acceptable salt or a diagnostically acceptable salt. A pharmaceutically or diagnostically acceptable salt refers to any organic or inorganic addition salt of the base compound of the active substance, at a concentration that is relatively non-toxic and harmless to the patient, and whose side effects do not reduce the beneficial efficacy of the base compound of the active substance. The free acid forming the acid addition salt can be an inorganic acid or an organic acid. Inorganic acids can include: hydrochloric acid, bromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc.; organic acids can include: citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, mesylic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, pamoic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, mesylic acid, ethanesulfonic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid, or malonic acid, etc. Furthermore, the salts include alkali metal salts (sodium salts, potassium salts, etc.) and alkaline earth metal salts (calcium salts, magnesium salts, etc.). For example, acid addition salts may include acetates, aspartates, benzoates, benzenesulfonates, bicarbonates / carbonates, bisulfates / sulfates, borates, camphorates, citrates, ethanedisulfonates, ethanesulfonates, formates, fumarates, gluconate, gluconate, glucuronates, hexafluorophosphates, hydroxybenzoylbenzene salts, hydrochlorides / chlorides, hydrobromates / bromines, hydroiodides / iodides, hydroxyethanesulfonates, lactates, malates, maleates, malonates, and so on. Sulfonates, methyl sulfates, naphthalates, 2-naphthalenesulfonates, nicotinates, nitrates, orotates, oxalates, palmitates, barmonates, phosphates / hydrogen phosphates / dihydrogen phosphates, glycosides, stearates, succinates, tartrates, toluenesulfonates, trifluoroacetates, aluminum, arginine, benzathine penicillin, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, zinc salts, etc., specifically hydrochloride or trifluoroacetate.

[0134] The acid addition salt can be prepared by conventional methods, such as dissolving the active substance in organic solvents such as methanol, ethanol, acetone, dichloromethane, and acetonitrile, then filtering and drying the precipitate obtained by adding organic or inorganic acids, or by distilling the solvent and excess acid under reduced pressure and then drying, or by crystallizing in an organic solvent.

[0135] In addition, metal salts can also be prepared using bases. For example, alkali metal or alkaline earth metal salts can be obtained by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved salt of the compound, and then evaporating and drying the filtrate. In this case, from a pharmaceutical perspective, it is suitable to produce sodium, potassium, or calcium salts in the form of metal salts. Furthermore, corresponding silver salts can be prepared by reacting alkali metal or alkaline earth metal salts with suitable silver salts (e.g., silver nitrate).

[0136] Furthermore, the compound represented by chemical formula 1 or its salt may include all solvates, hydrates and isomers that can be prepared therefrom.

[0137] The term "isomer" refers to compounds that have the same chemical or molecular formula but different structures or spatial arrangements. Examples of isomers can be structural isomers (such as tautomers), stereoisomers (such as R or S isomers with an asymmetric carbon center, geometric isomers (trans, cis), etc.), and optical isomers.

[0138] On the other hand, the present invention provides a method for preparing a compound represented by the following chemical formula 1, as shown in the following reaction formula 1, the method comprising the following steps:

[0139] Compound 1001 and compound 1002 were reacted in an organic solvent to obtain compound 1003 (step 1).

[0140] Compound 1003 and NH4OAc were reacted in an organic solvent to obtain compound 1004 (step 2).

[0141] Compound 1005-1, Compound 1005-2 or Compound 1005-3, and Compound 1004 are reacted in an organic solvent to obtain Compound 1006 (Step 3); and Compound 1006 is reacted with a catalyst in an organic solvent to obtain the compound represented by the following chemical formula 1 (Compound 1007) (Step 4).

[0142] [Reaction Formula 1]

[0143]

[0144] In the reaction formula 1,

[0145] The R is R 1c or X q ;

[0146] The X p and X q Each can be independently hydrogen, deuterium, or halogen;

[0147] The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0148] The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or

[0149] The R 1c and R 1d They connect to form the compound represented by the following chemical formula 2.

[0150] [Chemical Formula 2]

[0151]

[0152] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus,

[0153] The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0154] The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl;

[0155] The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

[0156] The terms “alkyl,” “alkoxy,” “aryl,” “heteroaryl,” and “salt,” etc., may fall within the above scope.

[0157] In one specific example, after step 4, the preparation method may further include the step of reacting the compound of chemical formula 1 with sulfuric acid. For example, a reaction as shown in the following reaction formula 2 can be carried out:

[0158] [Reaction 2]

[0159] .

[0160] In one specific example, the preparation method may further include the following step: after reacting compound 1004 and compound 1005-2 in an organic solvent in step 3, reacting an oxidant with an intermediate generated prior to obtaining compound 1006. For example, a reaction as shown in reaction formula 3 can be performed:

[0161] [Reaction 3]

[0162] .

[0163] In one specific example, the preparation method may further include the following step: after reacting compound 1004 and compound 1005-3 in an organic solvent in step 3, reacting an oxidant with an intermediate previously generated to obtain compound 1006. For example, a reaction as shown in the following reaction formula 4 can be performed:

[0164] [Reaction 4]

[0165] .

[0166] In one specific example, the oxidant may be one or more selected from the group consisting of pyridinium chlorochromate (PCC), pyridinium dichromate (PDC), Dess–Martin periodinane (DMP), 2-iodoxybenzoic acid (IBX), Swern oxidant, and Parikh-Doering oxidant.

[0167] In a specific example, when R is X q The preparation method may further include the following steps: after reacting compound 1006 with the catalyst in an organic solvent in step 4, reacting the Stille coupling starting material with the intermediate generated before obtaining the compound represented by formula 1. Specifically, the Stille coupling starting material may be (tributylstannyl)methanol.

[0168] In one specific example, the following steps may be further included: when R is X q At that time, after reacting with the Stille coupling reaction initiator, it reacts with R 1c (OTs) can react. For example, a reaction as shown in the following reaction formula 5 can be carried out:

[0169] [Reaction 5]

[0170] .

[0171] In one specific example, the organic solvents in steps 1 to 4 may be independently selected from one or more of the group consisting of methanol, ethanol, propanol, acetonitrile, dichloroethane (DCE), ethyl acetate, dimethyl sulfoxide, and dimethylformamide.

[0172] In one specific example, the catalyst in step 4 may be one or more selected from the group consisting of trifluoromethanesulfonic acid (TfOH), methanesulfonic acid (MsOH), p-toluenesulfonic acid (p-TsOH), and scandium triflate (Sc(OTf)3), but is not limited thereto.

[0173] On the other hand, a composition for detecting β-amyloid (Aβ) is provided, comprising a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0174] The term "β-amyloid protein" or "β-amyloid protein" refers to proteins that are intertwined and aggregated in the form of β-sheet structures, and refers to proteins or fragments capable of forming amyloid protein deposits. In this invention, β-amyloid protein can refer to a single protein constituting an aggregate of β-amyloid protein.

[0175] In one specific example, the β-amyloid protein may include aggregates of β-amyloid proteins, such as oligomers, protofibrils, fibrils, plaques, protein monomers and aggregates bound to homeobox proteins. Specifically, the β-amyloid protein may be one or more selected from the group consisting of Aβ oligomers, Aβ protofibrils, Aβ fibrils, and Aβ plaques. More specifically, the β-amyloid protein may be an Aβ oligomer.

[0176] In one specific example, the β-amyloid protein may have been mutated.

[0177] In one specific example, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may interact with one or more domains selected from the group consisting of: a domain containing the amino acid sequence of SEQ ID NO. 1, a domain containing the amino acid sequence of SEQ ID NO. 2, and a domain containing the amino acid sequence of SEQ ID NO. 3.

[0178] The term "domain" refers to the domain that makes up β-amyloid (Aβ). 1-42 In the amino acid sequence of a protein, the amino acid sequence or structural region to which a specific compound binds.

[0179] Specifically, the "structural domain containing the amino acid sequence of SEQ ID NO. n (where n is 1, 2 or 3)" may further include 0 to 5 (0, 1, 2, 3, 4 or 5) amino acid sequences starting from the N-terminal amino acid and / or C-terminal amino acid sequence of the amino acid.

[0180] For example, a domain containing the amino acid sequence of SEQ ID NO.1 may be a domain composed of SEQ ID NO.1, or a domain composed of the amino acid sequence of SEQ ID NO.1 and a sequence in which 1 to 5 amino acids are added to the N-terminus of the amino acid in SEQ ID NO.1, or a domain composed of the amino acid sequence of SEQ ID NO.1 and a sequence in which 1 to 5 amino acids are added to the C-terminus of the amino acid in SEQ ID NO.1, or a domain composed of the amino acid sequence of SEQ ID NO.1 and a sequence in which 1 to 5 amino acids are added to both the N-terminus and C-terminus of the amino acid in SEQ ID NO.1.

[0181] If the domain contains the amino acid sequence of SEQ ID NO.2, the domain may be a domain composed of SEQ ID NO.2, or a domain composed of the amino acid sequence of SEQ ID NO.2 and a sequence in which 1 to 5 amino acids are added to the N-terminus of the amino acid of SEQ ID NO.2.

[0182] Furthermore, for the structural domain containing the amino acid sequence of SEQ ID NO.3, the structural domain may be a structural domain composed of SEQ ID NO.3, or it may be a structural domain composed of the amino acid sequence of SEQ ID NO.3 and a sequence in which 1 to 5 amino acids are added to the C-terminus of the amino acid of SEQ ID NO.3.

[0183] The amino acid sequences of SEQ ID NO.1 to SEQ ID NO.3 are as follows:

[0184] SEQ ID NO.1: FFAEDVG, SEQ ID NO.2: MVGGVVIA, SEQ ID NO.3: DAEFRHDS.

[0185] In one specific example, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof interacts with one or more domains selected from the group consisting of: a domain consisting of an amino acid sequence having about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 92% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more sequence homology with the amino acid sequence of SEQ ID NO. 1; a domain consisting of an amino acid sequence having about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 92% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more sequence homology with the amino acid sequence of SEQ ID NO. 2; and a domain consisting of an amino acid sequence having about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 92% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more sequence homology with the amino acid sequence of SEQ ID NO. 1 ... The amino acid sequence of NO.3 has a domain consisting of amino acid sequences with approximately 70% or higher, approximately 75% or higher, approximately 80% or higher, approximately 85% or higher, approximately 90% or higher, approximately 92% or higher, approximately 95% or higher, approximately 97% or higher, approximately 98% or higher, or approximately 99% or higher sequence homology.

[0186] In one specific example, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may interact with one or more domains selected from the group consisting of: a domain consisting of the amino acid sequence of SEQ ID NO. 1, a domain consisting of the amino acid sequence of SEQ ID NO. 2, and a domain consisting of the amino acid sequence of SEQ ID NO. 3.

[0187] The composition for detection can detect β-amyloid protein by binding the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to an indicator. The indicator can be used to measure the degree of binding between β-amyloid protein and the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof; examples of the indicator may be fluorescent substances or fluorescent protein fragments.

[0188] The fluorescent substance may be a fluorescent dye with a basic backbone of rhodamine, coumarin, EvoBlue, oxazine, carbocyanine, naphthalene, biphenyl, anthracene, phenanthrene, pyrene or carbazole, or a derivative thereof. Specifically, the fluorescent substance may be selected from the group consisting of the following substances: Fluorescein, CR110: Rhodamine 110: Rhodamine Green (trade name), TAMRA: CarboxytetramethylRhodamine: TMR, Carboxyrhodamine 6G: CR6G, BODIPY FL (trade name): 4,4-difluoro-5,7-dimethyl-4-boron-3a,4a-diaza-s-indarsen-3-propionic acid, BODIPY 493 / 503 (trade name): 4,4-difluoro-1,3,5,7-tetramethyl-4-boron-3a,4a-diaza-s-indarsen-8-propionic acid, BODIPY R6G (trade name): 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-boron-3a,4a-diaza-s-indarsen-3-propionic acid, BODIPY 558 / 568 (trade name): 4,4-difluoro-5-(2-thienyl)-4-boron-3a,4a-diaza-s-indarsen-3-propionic acid; BODIPY 564 / 570 (trade name): 4,4-difluoro-5-styryl-4-boron-3a,4a-diaza-s-indarsen-3-propionic acid; BODIPY 576 / 589 (trade name): 4,4-difluoro-5-(2-pyrrole)-4-boron-3a,4a-diaza-s-indarsen-3-propionic acid; BODIPY 581 / 591 (trade name): 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-boron-3a.4a-diaza-s-indarin-3-propionic acid, EvoBlue10, EvoBlue30, MR121, ATTO 655, ATTO 680, ATTO 700, ATTO MB2, Alexa Fluor 350, Alexa Fluor 405, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 680, Alexa Fluor 700, Alexa Fluor 750, Alexa Fluor 790, Flamma 496, Flamma 507 (trade name), Flamma 530 (trade name), Flamma 552 (trade name), Flamma 560 (trade name), Flamma 575 (trade name), Flamma 581 (trade name), Flamma 648 (trade name), Flamma 675 (trade name), Flamma 749 (trade name), Flamma 774 (trade name), Flamma 775 (trade name), Rhodamine Red-X (trade name), Texas Red-X (trade name), 5(6)-TAMRA-X (trade name), 5-Carboxytetramethylrhodamine (5TAMRA) (trade name), Indocyanine Green (Indocyanine) green,ICG) and 2-((E)-2-((E)-2-(4-(2-carboxyethyl)phenoxy)-3-((E)-2-(3,3-dimethyl-5-sulfonato-1-(3-(tri-methylammonio)-propyl)indolin-2-ylidene)ethylidene)cyclohex-1-enyl)vinyl)-3,3-dimethyl-1-(3-(tri-methylammonio)-propyl)-3H-indolin-5-sulfonic acid sodium dibromide (2-((E)-2-((E)-2-(4-(2-carboxyethyl)phenoxy)-3-((E)-2-(3,3-dimethyl-5-sulfonato-1-(3-(tri-methyl ammonio)-propyl)indolin-2-ylidene)ethylidene)cyclohex-1-enyl)vinyl)-3,3-dimethyl-1-(3-(trimethyl ammonio)-propyl)-3H-indolium-5-sulfonate disodium bromide) (ZW800-1). ,

[0189] The fluorescent protein fragments include venus, Cerulean, Citrine, and mKate, which can be fluorescent proteins of different colors or partial structures thereof.

[0190] In one specific example, the composition used for detection can be used to confirm whether β-amyloid protein is detected in a sample isolated from an individual, and the isolated sample can be a biologically isolated sample.

[0191] The individual may be a mammal, such as a mouse, human, pig, cow, horse or sheep, and more specifically, a person suspected of having a disease mediated by the accumulation of β-amyloid protein.

[0192] The biologically isolated sample can be isolated from tissues where β-amyloid protein is distributed, specifically, it can be selected from one or more of the group consisting of blood, plasma, cerebrospinal fluid, brain tissue lysates, cortical lysates, and hippocampal lysates.

[0193] Another approach provides a method for detecting β-amyloid protein (Aβ), comprising the steps of contacting an isolated biological sample with a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0194] The method for detecting β-amyloid protein (Aβ) may further include the following steps: confirming the presence of β-amyloid protein or quantifying the amount of β-amyloid protein in a biological sample treated with the compound represented by the chemical formula 1 or its pharmaceutically acceptable salt.

[0195] The steps for confirming the presence or quantifying the amount of β-amyloid protein may include: confirming the binding of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof to the β-amyloid protein, and confirming the binding by measuring the emission spectrum emitted after the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof binds to the β-amyloid protein. Specifically, when an excitation source is irradiated into a biological sample treated with the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof, the presence or amount of β-amyloid protein can be analyzed by measuring the fluorescence produced by the compound represented by Chemical Formula 1 or its salt bound to the β-amyloid protein.

[0196] In one specific example, a microplate reader, spectrophotometer, microscope-based fluorescence detector, or flow cytometer can be used to measure the fluorescence signal. Alternatively, measurements can be performed using commercially available or known fluorescence detection systems, but are not limited to these.

[0197] The terms “pharmaceutically acceptable salt” and “β-amyloid protein” fall within the above scope.

[0198] On the other hand, a composition for diagnosing neurodegenerative diseases is provided, comprising a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0199] The term "neurodegenerative disease" refers to all diseases associated with degenerative changes in the brain, particularly all brain diseases caused by factors such as the aggregation and accumulation of β-amyloid protein in brain and / or brain nerve cells. Furthermore, some prodromal symptoms not considered as diseases are also included in the aforementioned β-amyloid-related diseases. Examples of neurodegenerative diseases may be selected from the group consisting of: mild cognitive impairment, dementia, Alzheimer's disease, hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D), preclinical Alzheimer's disease, Parkinson's disease, Huntington's disease, mild cognitive impairment, mild cognitive impairment due to Alzheimer's disease, and cerebral amyloid angiopathy. Angiopathy, Down syndrome, tauopathy, stroke due to amyloidosis, Luggage disease, systemic amyloidosis, Dutch amyloidosis, Niemann-Pick disease, Alzheimer's disease, amyotrophic lateral sclerosis, spinocerebellar atrophy, Tourette's syndrome, Friedrich's ataxia, Machado-Joseph's disease, Lewy body dementia, dystonia, progressive supranuclear palsy, and frontotemporal dementia.

[0200] The term "Alzheimer's disease" and "senile dementia" are used interchangeably. It is a disease accompanied by mental decline associated with a specific neurodegenerative disorder characterized by senile plaques, neuroinflammatory tangles, and progressive neuronal damage.

[0201] The term "Parkinson's disease" refers to a chronic and progressive degenerative disease of the central nervous system, whose most common symptoms are impaired motor function and language ability.

[0202] The term "huntington disease" refers to a neurodegenerative disease caused by the amplification of trinucleotide repeats in the gene encoding the huntingtin protein, accompanied by symptoms such as chorea, mental abnormalities, and dementia.

[0203] The term "Lugrigian disease" refers to a disease that selectively kills motor neurons, with symptoms including the gradual destruction of upper motor neurons in the cerebral cortex and lower motor neurons in the brainstem and spinal cord.

[0204] The term "Niemann-Pick disease" refers to a condition characterized by progressive damage to neurons in the brain.

[0205] The term "tau protein disease" can be a neurodegenerative disease in which neurons are damaged due to the abnormal accumulation of tau proteins (a family of proteins closely associated with intracellular microtubules) in brain tissue.

[0206] Compared to individuals without the disease, individuals with the neurodegenerative disease may have higher levels of β-amyloid protein. Specifically, the β-amyloid protein may be one or more selected from the group consisting of Aβ oligomers, Aβ fibrils, Aβ filaments, and Aβ plaques. More specifically, the β-amyloid protein may be Aβ oligomers. Furthermore, high levels of β-amyloid protein may be present in one or more tissues selected from the group consisting of plasma, blood, cerebrospinal fluid, cortex, and hippocampus.

[0207] The terms “pharmaceutically acceptable salt” and “β-amyloid protein” fall within the above scope.

[0208] Another approach provides a method for diagnosing neurodegenerative diseases, comprising the steps of contacting a biological sample with a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0209] The terms "pharmaceutically acceptable salt" and "neurodegenerative disease" both fall within the above scope.

[0210] Another aspect provides a method for providing information on the diagnosis of diseases associated with β-amyloid (e.g., neurodegenerative diseases), comprising the step of contacting an isolated biological sample with a compound represented by said chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0211] The method of providing information may include the following steps: confirming the presence of β-amyloid protein or quantifying the amount of β-amyloid protein in a biological sample that has come into contact with the compound represented by the chemical formula 1 or its pharmaceutically acceptable salt.

[0212] Furthermore, the method of providing information may further include the following steps: comparing the amount of β-amyloid protein obtained quantitatively with the amount of β-amyloid protein in a biological sample isolated from an individual who has never had a neurodegenerative disease (hereinafter referred to as the normal control group); and when the amount of β-amyloid protein obtained quantitatively is greater than the amount of β-amyloid protein in the normal control group, the method of providing information may further include determining that the individual has a neurodegenerative disease.

[0213] The terms “pharmaceutically acceptable salt,” “diseases associated with β-amyloid,” “neurodegenerative diseases,” and “β-amyloid” all fall within the above scope.

[0214] Another approach provides a method for screening β-amyloid aggregation inhibitors or aggregation dissolving agents, comprising the steps of contacting a sample containing β-amyloid with a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof.

[0215] The sample containing β-amyloid protein may be, for example, plasma, blood, cerebrospinal fluid, cortical lysate, or hippocampal lysate from a model animal of neurodegenerative disease, or it may be a cultured cell from a model animal of neurodegenerative disease, or it may be a solution containing purified β-amyloid protein.

[0216] The screening method can be used to screen for therapeutic agents for diseases associated with β-amyloid or neurodegenerative diseases. For example, a test substance and a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof can be administered to Alzheimer's disease model animals (in vivo); or a sample from an Alzheimer's disease model animal (ex vivo) or a test tube containing β-amyloid protein (in vitro) can be treated with the test substance and a compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof, and the amount of β-amyloid protein can be quantified. If the quantification of β-amyloid protein in the experimental group treated with the specific test substance is lower than that in the control group not treated with the specific test substance, the test substance can be selected as a disease therapeutic agent.

[0217] The terms “β-amyloid protein” and “pharmaceuticalally acceptable salt” fall within the above scope.

[0218] Beneficial effects

[0219] The compound represented by chemical formula 1 according to the present invention, or a salt thereof, can specifically bind to β-amyloid protein (Aβ). The compound, or a pharmaceutically acceptable salt thereof, due to its high selectivity, can detect β-amyloid protein in plasma with high sensitivity even if the presence of other proteins in the plasma obstructs detection, and can be used for the diagnosis of neurodegenerative diseases or for screening therapeutic agents for neurodegenerative diseases.

[0220] Brief description of the attached diagram

[0221] Figures 1A to 1F This is a graph showing the fluorescence intensity measured based on the binding of a compound to an Aβ monomer (mAβ), an Aβ oligomer (oAβ), or an Aβ filament (fAβ). Figure 1A and 1B IK15 compounds; Figures 1C to 1F :AMD compound).

[0222] Figure 2A and 2B It is a graph showing the fluorescence intensity over time after a compound is bound to an Aβ monomer (mAβ), an Aβ oligomer (oAβ), or an Aβ filament (fAβ).

[0223] Figure 3A It shows the fluorescence intensity of the combination of a compound with Aβ monomers (mAβ), Aβ oligomers (oAβ), and Aβ filaments (fAβ).

[0224] Figure 3B It is a graph showing the fluorescence intensity over time after a compound is bound to an Aβ monomer (mAβ), an Aβ oligomer (oAβ), or an Aβ filament (fAβ).

[0225] Figure 3C This is a diagram showing the binding of a compound to an Aβ oligomer (oAβ).

[0226] Figure 3D This is a diagram showing the Aβ binding sites of compounds confirmed according to one aspect.

[0227] Figure 3E This demonstrates confirmation based on one aspect of the compound and tau aggregates or Aβ oligomers (Aβ... 1-42 A diagram combining ).

[0228] Figure 3F This is a schematic diagram of an experiment used to identify the Aβ oligomer target sites of compounds according to one aspect.

[0229] Figures 3G to 3J This is a diagram showing the binding affinity of a compound to an Aβ fragment or Aβ complex, based on one aspect.

[0230] Figure 4A This is a schematic diagram of an experiment used to confirm the efficacy of Aβ based on the detection of one of the compounds in a 5XFAD transgenic mouse model.

[0231] Figure 4B The figure shows whether Aβ contained in the cerebral cortex lysate of a 5XFAD transgenic mouse model was detected according to the IK15j compound.

[0232] Figure 4C The figure shows whether Aβ contained in the hippocampal lysate of the 5XFAD transgenic mouse model was detected according to the IK15j compound of one side.

[0233] Figure 4D The figure shows whether Aβ contained in the cerebrospinal fluid (hereinafter referred to as CSF) of the 5XFAD transgenic mouse model was detected according to the IK15j compound of one side.

[0234] Figure 4E The figure shows whether Aβ contained in the plasma of a 5XFAD transgenic mouse model was detected based on the IK15j compound.

[0235] Figure 4F This is a schematic diagram showing the brain regions of mice treated with immunofluorescence according to a compound and an anti-Aβ antibody.

[0236] Figure 4G This is a diagram showing the immunofluorescence treatment of the brain of a 5XFAD transgenic mouse model using a compound according to one aspect and an anti-Aβ antibody.

[0237] Figure 5A and Figure 5B This is a diagram showing whether Aβ contained in the cerebral cortex lysate of a 5XFAD transgenic mouse model was detected based on one of the compounds.

[0238] Figure 6A and Figure 6B The figure shows whether Aβ contained in the hippocampal lysate of the 5XFAD transgenic mouse model was detected based on one of the compounds.

[0239] Figure 7A and Figure 7B This is a diagram showing whether Aβ contained in the cerebrospinal fluid of a 5XFAD transgenic mouse model was detected based on one of the compounds.

[0240] Figure 8The figure shows the confirmation of whether Aβ contained in the plasma of a 5XFAD transgenic mouse model was detected based on one of the compounds.

[0241] Figure 9A This is a graph showing the sensitivity of a compound to Aβ oligomers based on one aspect.

[0242] Figure 9B This is a diagram showing the binding of a compound to fibrinogen or albumin.

[0243] Figure 9C This is a diagram showing the binding of a compound, based on one aspect, to Aβ oligomers in the presence of albumin.

[0244] Figure 9D This is a schematic diagram showing an experiment used to confirm the binding of a compound according to one aspect to Aβ oligomers in the presence of albumin.

[0245] Figure 9E This is a diagram showing the binding of a compound, based on one aspect, to Aβ oligomers in the presence of albumin. Detailed Implementation

[0246] The present invention will now be described in more detail through the following embodiments. However, the following embodiments are merely illustrative of the invention, and the scope of the invention is not limited to these embodiments.

[0247] <Reference Example>

[0248] <Reference Example 1> Fluorescence Spectroscopy Scan

[0249] Fluorescence spectroscopy of IK15 was performed using a SpectraMax M5 molecular device. For excitation scanning, the compounds of this invention were dissolved in DMSO to prepare a 1 mM stock solution, which was then serially diluted with deionized water. 150 μL of the sample (100 μM) was then aliquoted into 96-well transparent round plates, and excitation spectral data points of the compounds were measured in increments of 5 nm from 200 nm to 900 nm. The peak value of each compound was recorded at the excitation wavelength. For emission scanning, 150 μL of the same sample prepared above was transferred to 96-well opaque round plates. The excitation wavelength was set according to the compound under study, and emission was scanned in 5 nm increments across various spectral ranges. Next, the emission spectra of the candidate compounds mixed with Aβ were measured to determine whether these compounds exhibited a fluorescence shift in the presence of Aβ.

[0250] <Reference Example 2> Preparation of MAP

[0251] A method for studying the compound of the present invention in Aβ was prepared with reference to Korean Patent Application No. 10-2023-0133625. 1-42 Fragment or Aβ 1-42 Mapping amyloid plate (MAP) for targeting sites in oligomers, the entire contents of which are incorporated herein by reference.

[0252] To remove bovine serum albumin coated on the culture plate, simply wash each well three times with 200 μL of washing buffer (0.1 M sodium phosphate, 0.15 M sodium chloride, 0.05% Tween-20, pH 7.2). Add Aβ... 1-42 Full-length and fragmented peptides were dissolved separately in DMSO to prepare 1.0 M stock solutions, which were then diluted with binding buffer (0.1 M sodium phosphate, 0.15 M sodium chloride, 10 mM EDTA, pH 7.2) to prepare 50 μg / mL peptide solutions (5% DMSO). 100 μL of the peptide solution was added to each well and reacted with maleimide overnight at room temperature. After peptide fixation, unbound peptides were washed three times with 200 μL of wash buffer. To inactivate residual maleimide groups, 200 μL of cysteine ​​solution (containing 10 μg / mL of binding buffer) was added to each well and incubated at room temperature for 1 hour. After capping with cysteine, all wells of the culture plate were washed three times with 200 μL of wash buffer.

[0253] <Reference Example 3> Study on the effect of IK15 on Aβ 1-42 Oligomer target sites

[0254] To confirm whether the compound of this invention targets the structure or sequence of the Aβ oligomer, two conditions were prepared. First, the compound of this invention (50 μM) was prepared in binding buffer (10% DMSO), and 100 μL of the compound was added to MAP and treated at room temperature for 24 hours. After incubation, all wells were washed three times with 200 μL of washing buffer. Second, the full-length Aβ oligomer was... 1-42 The peptide was added to the MAP and incubated at 37°C for 8 hours to form full-length Aβ. 1-42 —Aβ 1-42 Fragment complex. After incubation, all wells were washed three times with 200 μL of wash buffer and treated in MAP with 100 μL of the compound of the present invention (50 μM) for 24 hours at room temperature. After treatment, all wells were washed three times with 200 μL of wash buffer. The fluorescence intensity of the two culture plates was measured using a microplate reader (λex = 285 nm / λem = 475 nm).

[0255] <Reference Example 4> Animals

[0256] To investigate the interaction between the compounds of this invention and Aβ oligomers in brain lysate, cerebrospinal fluid (CSF), and plasma in an AD mouse model, 6-month-old female 5XFAD transgenic mice (strain name: B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas / -Mmjax expressing Swedish (K670N / M671L), Florida (I716V), and London (V717I) mutations of APP, as well as M146L and L286V mutations of PSEN1) and wild-type mice (C57BL / 6 xSJL) were obtained from Jackson Laboratory (Bar Harbor, Maine, USA). All animals were handled in accordance with the National Institutes of Health (NIH) Guidelines for the Household and Use of Laboratory Animals. The research protocol has been approved by the Yonsei University Animal Care and Use Committee (Seoul, South Korea, IACUC-202103-1221-01). All animal studies were conducted in accordance with the ARRIVE reporting guidelines.

[0257] <Reference Example 5> Analysis of brain tissue lysates

[0258] To prepare brain tissue lysates, mice were sacrificed, and the hippocampus and cortex regions of the mouse brain were dissected separately. Each brain region was homogenized in ice-cold RIPA buffer (20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 0.5% NP-40, 4 mM EDTA, 0.1% SDS, 0.5% sodium deoxycholate) containing a 1X protease inhibitor mixture (Roche Diagnostics, Switzerland). The homogenized brain tissue was incubated on ice for 20 min, then centrifuged at 14,000 rpm for 30 min at 4°C, and the supernatant (soluble fraction) of the brain tissue lysates was collected. To analyze the interaction between the compound of the present invention and mouse brain tissue lysate samples, diluted brain tissue lysate samples were mixed with the compound of the present invention (50 μM). The lysate samples containing the compound were loaded into the wells of 96-well half-region black microplates (Corning Electron, USA). The samples were distributed and detected at λex = 285 nm / λem = 475 nm.

[0259] <Reference Example 6> Analysis of Cerebrospinal Fluid (CSF)

[0260] To analyze the interaction between the compound of the present invention and mouse cerebrospinal fluid (CSF), CSF samples from euthanized mice were used. Each CSF sample was treated with the compound of the present invention (50 μM). Individual samples and the compound of the present invention were added to the wells of a 96-well half-zone black microplate. The total volume of each sample was 50 μL. The signal of each sample was detected using a microplate reader (λex = 285 nm / λem = 475 nm).

[0261] <Reference Example 7> Analysis of plasma

[0262] To analyze the interaction between the compounds of this invention and mouse plasma, plasma samples from euthanized mice were used. Individual samples and the compounds of this invention were added to the wells of a 96-well half-zone black microplate. The total volume of each sample was 50 μL. The signal of each sample was detected using a microplate reader (λex = 285 nm / λem = 475 nm).

[0263] <Reference Example 8> Staining brain tissue from an AD mouse model

[0264] Mouse brain tissue was fixed overnight in 4% paraformaldehyde (Biosesang, Korea) at 4°C, and then cryoprotected for 48 hours in 30% sucrose. Brain tissue sections (25 μm) were cut using a cryostat (CM1860, Leica) and adhered to glass slides. The compound of this invention (500 μM) was treated with the brain tissue overnight at room temperature. After washing with 1X PBS, 1% SDS (Biosesang, Korea) was added to 1X PBS (Gibco, Korea) for antigen retrieval of the fixed brain tissue sections for 10 minutes, followed by blocking with 20% horse serum in 1X PBS. After 1 hour, the slides were incubated with mouse monoclonal antibody 6E10 (1:200, BioLegend, USA) at room temperature for 1 hour. After washing with 1X PBS, the slides were incubated with goat anti-mouse IgG labeled with Alexa Fluor 555 (1:200, Invitrogen, USA) for 1 hour at room temperature. As a positional marker, brain tissue sections were stained with Hoechst 33342 (10 μg / mL in PBS) for 3 minutes at room temperature. After staining the cortical and hippocampal regions of the fixed brain tissue, each brain tissue section was visualized using a fluorescence microscope (DM500, Leica) along with a filter cube containing excitation and emission filters: the N2.1 filter cube was used to detect 6E10 staining (excitation filter: BP515-560; dichroic mirror: 580; emission filter: LP 590) and the L5 filter cube was used to detect the compounds of this invention (excitation filter: BP 480 / 40; dichroic mirror: 505; emission filter: BP 527 / 30).

[0265] <Reference Example 9> Plasma Aβ oligomers were detected.

[0266] Aβ 1-42 Peptides (1000, 100, 10, 1, 0.1, 0.01, 0.001 pg / mL) were aggregated at 37°C for 8 hours and then added to human plasma buffer (Sigma-Aldrich, USA). Each plasma sample was then treated with the compound of this invention (100 μM) at room temperature for 8 hours. The signal of each sample was detected using a microplate reader (λex = 285 nm / λem = 475 nm).

[0267] <Reference Example 10> Selectively study the effect of IK15 on blood proteins

[0268] Aβ 1-42 The peptides were aggregated at 37°C for 8 hours, and then 25 μM of Aβ was added. 1-42Oligomers, albumin (Sigma-Aldrich, USA), and fibrinogen (Sigma-Aldrich, USA) were seeded into black microplates. Each plasma sample was treated with IK15j (100 μM) at room temperature for 8 hours. The signal of each sample was detected using a microplate reader (λex = 285 nm / λem = 475 nm).

[0269] <Reference Example 11> Selectively study the effect of IK15 on the albumin-Aβ oligomer complex.

[0270] The full length Aβ 1-42 The peptide was immobilized on a maleimide-activated microplate. Then, additional Aβ was added. 1-42 The peptide (10 μM) was treated at 37 °C for 8 hours. After peptide fixation, unbound peptides were washed three times with 200 μL of washing buffer. To form the albumin-Aβ oligomer complex, albumin (10 μM, Sigma-Aldrich, USA) was treated on a culture plate at room temperature for 24 hours, followed by washing three times with 200 μL of washing buffer for unbound residues. The Aβ oligomer wells, serving as a control, were not treated with albumin. The compound of the present invention (100 μM) was applied to each well, and the signal for each sample was detected using a microplate reader (λex = 285 nm / λem = 475 nm).

[0271] <Reference Example 12> Statistical Analysis

[0272] All graphical data were analyzed using GraphPad Prism 9.0 software. Statistical analysis was performed using one-way ANOVA, followed by Bonferroni post-hoc comparisons or Student's unpaired t-test. Error bars represent the standard error (SEM) of the mean.

[0273] <Preparation Example>

[0274] <Preparation Example 1> Preparation Examples 1 to 13

[0275] Examples of synthesizing compounds according to one aspect according to the following reaction formula 6 (hereinafter referred to as Examples 1 to 13).

[0276] [Reaction Formula 6]

[0277] .

[0278] Several commercially available α-bromoketones were treated with NaCN to obtain compound 2001. Then, β-ketonitrile was alkylated with 2-chloroacetone in a base-mediated manner to give 1,4-diketo compound 2002. These were then treated with NH4OAc to convert them into the corresponding 2-arylpyrrole compound 2003. Subsequently, several α-haloketones and 2-arylpyrrole compound 2003 were N-alkylated to give compound 2004. Then, under optimized conditions, compound 2004 was synthesized into pyrrolo[2,1-a]isoquinoline compound 2005, which is a variety of N-substituted 2-arylpyrrole derivatives 2004. By modifying the α-bromoketone or the α-haloketone, compounds of 13 embodiments were prepared (Example 1 (the compound represented by chemical formula 3; IK15a), Example 2 (the compound represented by chemical formula 4; IK15b), Example 3 (the compound represented by chemical formula 5; IK15c), Example 4 (the compound represented by chemical formula 6; IK15d), Example 5 (the compound represented by chemical formula 7; IK15e), Example 6 (the compound represented by chemical formula 8; IK15c), and Example 7 (the compound represented by chemical formula 8; IK15c). 15f), Example 7 (the compound represented by chemical formula 9; IK15l), Example 8 (the compound represented by chemical formula 10; IK15n), Example 9 (the compound represented by chemical formula 11; IK15o), Example 10 (the compound represented by chemical formula 12; IK15p), Example 11 (the compound represented by chemical formula 13; IK15q), Example 12 (the compound represented by chemical formula 14; IK15r), and Example 13 (the compound represented by chemical formula 15; IK15s) .

[0279] For the compounds synthesized in Examples 1 to 13, H-NMR (400 MHz, CDCl3) and C-NMR (100 MHz, CDCl3) analyses were performed according to known nuclear magnetic resonance spectroscopy methods to analyze the structure of the substances, and the results are as follows:

[0280] [Example 1 (The compound represented by the chemical formula 3; IK15a; 3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0281] 1 H NMR (400 MHz, CDCl3)δ 8.91(d, J = 8.0 Hz, 1H), 7.62 (s, 3H), 7.54-7.46 (m, 6H), 6.76 (s, 1H), 2.49 (s, 3H),

[0282] 13 C{1H} NMR (100 MHz, CDCl3) δ 136.5, 133.3, 130.2, 128.7, 128.3,128.2, 127.6, 127.5, 126.7, 125.9, 125.0, 123.6, 123.1, 120.1, 118.9, 114.3,82.4, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 20 H 15 N2 283.1230, found280.1239.

[0283] [Example 2 (the compound represented by the chemical formula 4; IK15b; 6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0284] 1 H NMR (400 MHz, CDCl3)δ 8.89 (d, J = 7.6 Hz, 1H), 7.67-7.62 (m, 2H), 7.59 (s, 1H), 7.50-7.45 (m, 1H), 7.40 (d, J = 7.6 Hz, 2H), 7.06 (d, J = 7.6Hz, 2H), 6.75 (s, 1H), 3.91 (s, 3H), 2.48 (s, 3H),

[0285] 13C{1H} NMR (100 MHz, CDCl3) δ 159.6, 133.2, 131.3, 128.6, 128.2,127.8, 127.6, 126.3, 125.9, 125.0, 123.5, 123.1, 120.0, 119.0, 114.2, 114.1,82.3, 55.4, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 21 H 17 N2O 313.1336, found 313.1340.

[0286] [Example 3 (The compound represented by the chemical formula 5; IK15c; 3-methyl-6-(p-tolyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0287] 1 H NMR (400 MHz, CDCl3)δ 8.90 (d, J = 7.6 Hz, 1H), 7.66-7.62 (m, 2H), 7.60 (s, 1H), 7.50-7.43 (m, 1H), 7.39-7.33 (m, 4H), 6.75 (s, 1H), 2.47 (s,6H),

[0288] 13 C{1H} NMR (100 MHz, CDCl3) δ 138.1, 133.5, 133.2, 130.0, 129.4,128.3, 127.7, 127.6, 126.6, 125.9, 125.0, 123.6, 123.1, 120.0, 119.0, 114.2,82.4, 21.3, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 21 H 17 N2 297.1386, found297.1392.

[0289] [Example 4 (the compound represented by the chemical formula 6; IK15d; 6-(4-chlorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0290] 1 H NMR (400 MHz, CDCl3)δ 8.89 (d, J = 8.0 Hz, 1H), 7.65-7.60 (m, 1H), 7.59 (s, 1H), 7.56 (s, 1H), 7.52-7.47 (m, 3H), 7.42 (d, J = 8.4 Hz, 2H), 6.76(s, 1H), 2.49 (s, 3H),

[0291] 13 C{1H} NMR (100 MHz, CDCl3) δ 134.9, 134.4, 133.2, 131.5, 129.0,128.5, 127.7, 127.2, 125.6, 125.5, 125.0, 123.7, 123.2, 120.2, 118.8, 114.4,82.7, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 20 H 14 N2Cl 317.0840, found317.0828.

[0292] [Example 5 (The compound represented by the chemical formula 7; IK15e; 3-methyl-6-(naphthalen-2-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0293] 1 H NMR (400 MHz, CDCl3)δ 8.93 (d, J = 8.0 Hz, 1H), 8.01-7.91 (m, 4H), 7.71 (s, 1H), 7.68-7.63 (m, 2H), 7.59 (s, 3H), 7.50-7.43 (m, 1H), 6.78 (s,1H), 2.51 (s, 3H),

[0294] 13 C{1H} NMR (100 MHz, CDCl3) δ 134.0, 133.4, 133.3, 132.9, 129.0,128.4, 128.2, 128.1, 128.0, 127.8, 127.7, 127.6, 126.69, 126.65, 126.6,126.0, 125.1, 123.7, 123.1, 120.4, 118.9, 114.3, 82.5, 11.6; HRMS (ESI-QTOF)m / z [M+H]+ calcd for C 24 H 17 N2 333.1386, found 333.1396.

[0295] [Example 6 (The compound represented by the chemical formula 8; IK15f; 3,6-dimethylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0296] 1 H NMR (400 MHz, CDCl3) δ 8.83 (d, J = 7.2 Hz, 1H), 7.77 (d, J = 6.8Hz, 1H), 7.63-7.56 (m, 2H), 7.51 (s, 1H), 6.67 (s, 1H), 2.50 (s, 3H), 2.46(s, 3H),

[0297] 13 C{1H} NMR (100 MHz, CDCl3) δ 133.2, 128.3, 128.0, 127.6, 124.8,123.8, 123.1, 123.0, 119.5, 119.2, 119.1, 113.6, 82.0, 16.7, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 15 H 13 N2 221.1073, found 221.1084.

[0298] [Example 7 (the compound represented by the chemical formula 9; IK15l; 5-(4-methoxyphenyl)-8-methylbenzo[f]pyrrolo[2,1-a]isoquinoline-10-carbonitrile)]

[0299] 1 H NMR (400 MHz, CDCl3) δ 8.95 (d, J = 7.2 Hz, 1H), 8.80 (d, J = 7.6Hz, 1H), 7.89 (d, J = 7.2 Hz, 1H), 7.65 (s, 2H), 7.45 (s, 1H), 7.34 (d, J =6.4 Hz, 2H), 7.15 (s, 1H), 7.04 (d, J = 6.4 Hz, 2H), 6.85 (s, 1H), 3.93 (s,3H), 2.50 (s, 3H),

[0300] 13 C{1H} NMR (100 MHz, CDCl3) δ 159.4, 133.6, 133.2, 130.4, 130.1,129.5, 128.8, 127.5, 126.2, 125.5, 124.5, 123.6, 122.6, 120.7, 119.3, 115.7,114.5, 81.7, 55.4, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 25 H 19 N2O363.1492, found 363.1502.

[0301] [Example 8 (the compound represented by the chemical formula 10; IK15n; 3-methyl-6,8-diphenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0302] 1H NMR (400 MHz, CDCl3) δ 8.97 (d, J = 8.4 Hz, 1H), 7.88 (d, J = 8.4Hz, 1H), 7.84 (s, 1H), 7.64 (s, 1H), 7.59-7.52 (m, 7H), 7.43 (t, J = 7.2 Hz,2H), 7.36 (d, J = 6.0 Hz, 1H), 6.78 (s, 1H), 2.50 (s, 3H),

[0303] 13 C{1H} NMR (100 MHz, CDCl3) δ 140.3, 135.4, 133.2, 130.1, 128.9,128.8, 128.3, 128.0, 127.7, 127.5, 127.2, 126.8, 124.1, 123.9, 123.8, 123.6,120.5, 118.9, 114.4, 93.3, 82.5, 11.6; HRMS (ESI-QTOF) m / z [M+H]+ calcd forC 26 H 19 N2 359.1543, found 359.2311.

[0304] [Example 9 (the compound represented by the chemical formula 11; IK15o; 6-(4-methoxyphenyl)-3-methyl-8-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0305] 1 H NMR (400 MHz, CDCl3) δ 8.94 (d, J = 8.4 Hz, 1H), 7.88-7.84 (m, 2H), 7.61 (s, 1H), 7.57 (d, J = 7.6 Hz, 2H), 7.46-7.41 (m, 4H), 7.37 (d, J = 6.8Hz, 1H), 7.06 (d, J = 8.4 Hz, 2H), 6.75 (s, 1H), 3.91 (s, 3H), 2.49 (s, 3H),

[0306] 13C{1H} NMR (100 MHz, CDCl3) δ 159.6, 140.3, 130.2, 133.1, 131.3,128.9, 128.6, 128.3, 127.7, 127.3, 127.2, 126.4, 124.03, 123.96, 123.7,123.6, 120.4, 119.0, 114.3, 114.2, 82.4, 55.4, 11.6; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 27 H 21 N2O 389.1649, found 389.1658.

[0307] [Example 10 (the compound represented by the chemical formula 12; IK15p; 3-methyl-8-phenyl-6-(p-tolyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0308] 1 H NMR (400 MHz, CDCl3) δ 8.96 (d, J = 8.8 Hz, 1H), 7.89-7.84 (m, 2H), 7.62 (s, 1H), 7.57 (d, J = 7.6 Hz, 2H), 7.46-7.41 (m, 3H), 7.40 (s, 1H),7.37-7.33 (m, 3H), 6.77 (s, 1H), 2.49 (s, 3H), 2.48 (s, 3H),

[0309] 13 C{1H} NMR (100 MHz, CDCl3) δ 140.4, 140.3, 138.1, 133.4, 133.1,130.0, 129.5, 128.9, 128.1, 127.7, 127.4, 127.3, 126.7, 124.1, 124.0, 123.7,123.6, 120.4, 119.0, 114.3, 82.4, 21.3, 11.6; HRMS (ESI-QTOF) m / z [M+H]+calcd for C 27 H 21 N2 373.1699, found 373.1693.

[0310] [Example 11 (The compound represented by the chemical formula 13; IK15q; 3,6-dimethyl-8-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0311] 1 H NMR (400 MHz, CDCl3) δ 8.87 (d, J = 8.4 Hz, 1H), 7.93 (s, 1H), 7.85 (d, J = 6.8 Hz, 1H), 7.70 (d, J = 6.8 Hz, 2H), 7.52 (d, J = 6.0 Hz, 2H), 7.49(s, 1H), 7.42 (d, J = 6.4 Hz, 1H), 6.67 (s, 1H), 2.54 (s, 3H), 2.47 (s, 3H),

[0312] 13 C{1H} NMR (100 MHz, CDCl3) δ 140.5, 140.3, 133.1, 129.0, 128.7,127.8, 127.3, 127.2, 123.9, 123.6, 131.2, 122.0, 119.7, 119.5, 119.1, 113.8,82.1, 16.8, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 21 H 17 N2 297.1386, found297.1391.

[0313] [Example 12 (the compound represented by the chemical formula 14; IK15r; 8-chloro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0314] 1H NMR (400 MHz, CDCl3) δ 8.80 (d, J = 6.4 Hz, 1H), 7.60 (d, J = 4.8Hz, 2H), 7.55 (d, J = 7.6 Hz, 1H), 7.38 (d, J = 6.0 Hz, 2H), 7.07 (d, J = 6.0Hz, 2H), 6.75 (s, 1H), 3.91 (s, 3H), 2.48 (s, 3H),

[0315] 13 C{1H} NMR (100 MHz, CDCl3) δ 159.8, 133.5, 132.5, 131.2, 129.3,128.6, 127.8, 125.4, 125.3, 124.5, 123.9, 123.4, 121.0, 118.6, 114.4, 114.3,82.7, 55.4, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 21 H 16 ClNO 347.0946, found 347.0952.

[0316] [Example 13 (the compound represented by the chemical formula 15; IK15s; 8-bromo-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile)]

[0317] 1 H NMR (400 MHz, CDCl3) δ 8.74 (d, J = 8.8 Hz, 1H), 7.75 (s, 1H), 7.69 (d, J = 7.2 Hz, 1H), 7.60 (s, 1H), 7.37 (d, J = 8.4 Hz, 2H), 7.07 (d, J = 8.0Hz, 2H), 6.76 (s, 1H), 3.91 (s, 3H), 2.48 (s, 3H),

[0318] 13C{1H} NMR (100 MHz, CDCl3) δ 159.8, 132.6, 131.3, 131.2, 129.5,128.4, 127.8, 125.3, 123.6, 123.9, 123.7, 121.7, 121.0, 118.6, 114.5, 114.3,82.9, 55.4, 11.5; HRMS (ESI-QTOF) m / z [M+H]+ calcd for C 21 H 16 BrN 2O 391.0441, found 391.0444.

[0319] <Preparation Example 2> Preparation Example 14 (the compound represented by chemical formula 16; AMD-D-253)

[0320] The compound of Example 14 was synthesized according to the following reaction formula 7.

[0321] [Reaction Formula 7]

[0322] .

[0323] Step 1. Preparation of 2-(4-fluorobenzoyl)-4-oxopentanenitrile

[0324] 3-(4-fluorophenyl)-3-oxopropionitrile (5 g, 30.60 mmol) and 1-chloropropane-2-one (3.1 g, 33.66 mmol) were mixed in ethanol (80 mL), potassium carbonate (10.5 g, 76.50 mmol) was added, and the mixture was stirred at 25 °C for 2 hours under a nitrogen atmosphere. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (120 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v = 5:95) to give 2-(4-fluorobenzoyl)-4-oxopentanonitrile (3 g, 44% yield).

[0325] Step 2. Preparation of 2-(4-fluorophenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0326] 2-(4-fluorobenzoyl)-4-oxopentanilonitrile (3 g, 13.70 mmol) and ammonium acetate (2.1 g, 27.40 mmol) were mixed in ethanol (50 mL) and stirred at 80 °C for 5 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 12:88) to give 2-(4-fluorophenyl)-5-methyl-1H-pyrrole-3-onitrile (2.2 g, 80% yield) as a yellow solid.

[0327] Step 3. Preparation of 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrole-3-carbonitrile

[0328] 2-(4-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.99 mmol) and 2-bromo-1-phenylethane-1-one (238 mg, 1.20 mmol) were mixed in acetonitrile (60 mL), and cesium carbonate (651 mg, 1.99 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (40 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (80 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (220 mg, 69% yield) as a white solid.

[0329] Step 4. Preparation of 8-fluoro-3-methyl-6-phenylpyrrolo[2,1-a] isoquinoline-1-carbonitrile (Example 14)

[0330] 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (200 mg, 0.63 mmol), trifluoromethanesulfonic acid (471 mg, 3.14 mmol), and 1,2-dichloroethane (30 mL) were mixed and stirred at 60 °C for 2 hours. Water (25 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by preparative high-performance liquid chromatography (Prep-HPLC) (25%-95% acetonitrile / 0.1% ammonia) to give 8-fluoro-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (19.8 mg, 10% yield) as a white solid.

[0331] LCMS (ESI-MS): Calculation of molecular weight (mass calcd) C 20 H 13 FN2 300.1 m / z, Found 301.1 [M+H]⁺.

[0332] 1 H NMR (400 MHz, CDCl3) δ: 8.91 (dd, J = 9.2, 5.6 Hz, 1H), 7.65 (s,1H), 7.57–7.50 (m, 3H), 7.49–7.45 (m, 2H), 7.39–7.33 (m, 1H), 7.30–7.27 (m,1H), 6.76 (s, 1H), 2.49 (s, 3H).

[0333] <Preparation Example 3> Preparation Example 15 (the compound represented by chemical formula 17; AMD-D-254)

[0334] The compound of Example 15 was synthesized according to the following reaction formula 8.

[0335] [Reaction Equation 8]

[0336] .

[0337] Step 1. Preparation of 2-(4-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0338] The intermediate 2-(4-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.99 mmol) from step 2 of Example 14 and 2-bromo-1-(4-methoxyphenyl)ethane-1-one (275 mg, 1.20 mmol) were mixed in acetonitrile (60 mL), and cesium carbonate (651 mg, 1.99 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (40 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (80 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(4-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (220 mg, yield 63%), which was a white solid.

[0339] Step 2. Preparation of 8-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a] isoquinoline-1-carbonitrile (Example 15)

[0340] 2-(4-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.29 mmol), trifluoromethanesulfonic acid (430 mg, 2.87 mmol), and 1,2-dichloroethane (30 mL) were mixed and stirred at 80 °C for 2 hours. Water (25 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by preparative high-performance liquid chromatography (Prep-HPLC) (25%–95% acetonitrile / 0.1% ammonia) to give 8-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (25.9 mg, 27% yield) as a white solid.

[0341] LCMS (ESI-MS): Calculation of molecular weight C 21 H 15 FN2O 330.1 m / z, observed value 330.9 [M+H]⁺.

[0342] 1H NMR (400 MHz, CDCl3) δ: 8.89 (dd, J = 8.8, 5.2 Hz, 1H), 7.62 (s,1H), 7.40–7.37 (m, 2H), 7.36–7.27 (m, 2H), 7.08–7.05 (m, 2H), 6.75 (s, 1H), 3.91 (s, 3H), 2.48 (s, 3H).

[0343] <Preparation Example 4> Preparation Example 16 (the compound represented by chemical formula 18; AMD-D-255)

[0344] The compound of Example 16 was synthesized according to the following reaction formula 9.

[0345] [Reaction Formula 9]

[0346] .

[0347] Step 1. Preparation of 2-(4-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0348] The intermediate 2-(4-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (300 mg, 1.50 mmol) from step 2 of Example 14 and 2-bromo-1-(4-fluorophenyl)ethane-1-one (390 mg, 1.80 mmol) were mixed in acetonitrile (40 mL), and cesium carbonate (976 mg, 3.00 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (40 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (80 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio = 10:90) to give 2-(4-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (150 mg, yield 29%), which was a white solid.

[0349] Step 2. Preparation of 8-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a] isoquinoline-1-carbonitrile (Example 16)

[0350] 2-(4-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.29 mmol), trifluoromethanesulfonic acid (223 mg, 1.49 mmol), and 1,2-dichloroethane (30 mL) were mixed and stirred at 80 °C for 1 hour. Water (25 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by preparative high-performance liquid chromatography (Prep-HPLC) (25%–65% acetonitrile / 0.1% ammonia) to give 8-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (20.1 mg, 21% yield) as a white solid.

[0351] LCMS (ESI-MS): Calculation of molecular weight C 20 H 12 F2N2 318.1 m / z, observed value 318.6 [M+H]⁺.

[0352] 1 H NMR (400 MHz, CDCl3) δ: 8.91 (dd, J = 9.2, 5.6 Hz, 1H), 7.63 (s,1H), 7.46–7.42 (m, 2H), 7.39–7.34 (m, 1H), 7.25–7.19 (m, 3H), 6.77 (s, 1H),2.49 (s, 3H).

[0353] <Preparation Example 5> Preparation Example 17 (the compound represented by chemical formula 19; AMD-D-257)

[0354] The compound of Example 17 was synthesized according to the following reaction formula 10.

[0355] [Reaction Formula 10]

[0356] .

[0357] Step 1. Preparation of 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-carbonitrile

[0358] The intermediate 2-(4-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (300 mg, 1.50 mmol) from step 2 of Example 14 and 2-bromo-1-(thiophen-2-yl)ethane-1-one (369 mg, 1.80 mmol) were mixed in acetonitrile (50 mL), and cesium carbonate (976 mg, 3.00 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (40 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (80 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio = 10:90) to give 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-onitrile (160 mg, yield 33%), which was a white solid.

[0359] Step 2. Preparation of 8-fluoro-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 17)

[0360] 2-(4-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrolo-3-onitrile (100 mg, 0.31 mmol), trifluoromethanesulfonic acid (231 mg, 1.54 mmol), and 1,2-dichloroethane (30 mL) were mixed and stirred at 80 °C for 2 hours. Water (25 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by preparative high-performance liquid chromatography (Prep-HPLC) (25%–70% acetonitrile / 0.1% ammonia) to give 8-fluoro-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-onitrile (AMD-D-257) (18.5 mg, 19% yield), as a white solid.

[0361] LCMS (ESI-MS): Calculation of molecular weight C 18 H 11 FN2S 306.1 m / z, observed value 307.0 [M+H]⁺.

[0362] 1 H NMR (400 MHz, CDCl3) δ: 8.89 (dd, J = 8.8, 5.2 Hz, 1H), 7.79 (s,1H), 7.58–7.56 (m, 1H), 7.51–7.48 (m, 1H), 7.41–7.35 (m, 1H), 7.25–7.21 (m,2H), 6.76 (s, 1H), 2.50 (s, 3H).

[0363] <Preparation Example 6> Preparation Example 18 (the compound represented by chemical formula 20; AMD-D-258)

[0364] The compound of Example 18 was synthesized according to the following reaction formula 11.

[0365] [Reaction Formula 11]

[0366] .

[0367] Step 1. Preparation of 4-oxo-2-(4-(trifluoromethyl)benzoyl)pentanenitrile

[0368] 3-Oxo-3-[4-(trifluoromethyl)phenyl]propionitrile (5 g, 0.023 mol) and potassium carbonate (8.12 g, 0.058 mol) were mixed in ethanol (150 mL), followed by the addition of 1-chloropropane-2-one (2.39 g, 0.025 mol). The mixture was stirred at 25 °C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 4-oxo-2-(4-(trifluoromethyl)benzoyl)pentanonitrile (5 g, 79% yield) as a yellow solid.

[0369] Step 2. Preparation of 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-carbonitrile

[0370] 4-Oxo-2-(4-(trifluoromethyl)benzoyl)pentanonitrile (2.5 g, 0.009 mol) and ammonium acetate (1.51 g, 0.019 mol) were mixed in ethanol (50 mL) and stirred at 80 °C for 2 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (1.1 g, 43% yield) as a white solid.

[0371] Step 3. Preparation of 5-methyl-1-(2-oxo-2-phenylethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-carbonitrile

[0372] A mixture of 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (200 mg, 0.799 mmol), 2-bromo-1-phenylethane-1-one (190.92 mg, 0.959 mmol), and acetonitrile (9 mL) was added, along with cesium carbonate (520.86 mg, 1.598 mmol). The mixture was stirred at 60 °C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 8:92) to give 5-methyl-1-(2-oxo-2-phenylethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (168 mg, 45% yield) as a yellow solid.

[0373] Step 4. Preparation of 3-methyl-6-phenyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 18)

[0374] A mixture of 5-methyl-1-(2-oxo-2-phenylethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (100 mg, 0.271 mmol), trifluoromethanesulfonic acid (407.47 mg, 2.715 mmol), and 1,2-dichloroethane (5 mL) was stirred at 60 °C for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) to give 3-methyl-6-phenyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-onitrile (27 mg, 27% yield) as a white solid.

[0375] LCMS (ESI-MS): Calculation of molecular weight C 21 H 13 F3N2 350.1 m / z, observed value 350.7 [M+H]⁺.

[0376] 1 H NMR (400 MHz, CDCl3) δ: 9.00 (d, J = 8.4 Hz, 1H), 7.89 (s, 1H), 7.84–7.81 (m, 1H), 7.70 (s, 1H), 7.58–7.51 (m, 3H), 7.48–7.46 (m, 2H), 6.84(s, 1H), 2.52(s, 3H).

[0377] <Preparation Example 7> Preparation Example 19 (the compound represented by chemical formula 21; AMD-D-259)

[0378] The compound of Example 19 was synthesized according to the following reaction formula 12.

[0379] [Reaction 12]

[0380] .

[0381] Step 1. Preparation of 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-carbonitrile)

[0382] The intermediate 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (200 mg, 0.799 mmol) from step 2 of Example 18 and 2-bromo-1-(4-methoxyphenyl)ethane-1-one (219.72 mg, 0.959 mmol) were mixed in acetonitrile (9 mL), and cesium carbonate (520.86 mg, 1.598 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 8:92) to give 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-onitrile (150 mg, yield 37%), which was a yellow solid.

[0383] Step 2. Preparation of 6-(4-methoxyphenyl)-3-methyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 19)

[0384] A mixture of 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (100 mg, 0.251 mmol), trifluoromethanesulfonic acid (376.7 mg, 2.51 mmol), and 1,2-dichloroethane (5 mL) was stirred at 60 °C for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) to give 6-(4-methoxyphenyl)-3-methyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-onitrile (14 mg, 14% yield) as a white solid.

[0385] LCMS (ESI-MS): Calculation of molecular weight C 22 H 15 F3N 2O 380.1 m / z, observed value 380.7 [M+H]⁺.

[0386] 1 H NMR (400 MHz, CDCl3) δ: 8.99 (d, J = 8.4 Hz, 1H), 7.91 (s, 1H), 7.83–7.80 (m, 1H), 7.67 (s, 1H), 7.43–7.36 (m, 2H), 7.12–7.05 (m, 2H), 6.83(s, 1H), 3.92(s, 3H), 2.51(s, 3H).

[0387] <Preparation Example 8> Preparation Example 20 (the compound represented by chemical formula 22; AMD-D-260)

[0388] The compound of Example 20 was synthesized according to the following reaction formula 13.

[0389] [Reaction Formula 13]

[0390] .

[0391] Step 1. Preparation of 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-carbonitrile)

[0392] The intermediate 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (200 mg, 0.799 mmol) from step 2 of Example 18 and 2-bromo-1-(4-fluorophenyl)ethane-1-one (208.17 mg, 0.959 mmol) were mixed in acetonitrile (9 mL), and cesium carbonate (520.86 mg, 1.598 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-onitrile (100 mg, yield 29%), which was a yellow solid.

[0393] Step 2. Preparation of 6-(4-fluorophenyl)-3-methyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 20)

[0394] A mixture of 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (100 mg, 0.258 mmol), trifluoromethanesulfonic acid (388.41 mg, 2.588 mmol), and 1,2-dichloroethane (5 mL) was stirred at 60 °C for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 6:94) to give 6-(4-fluorophenyl)-3-methyl-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-onitrile (10 mg, 10% yield) as a white solid.

[0395] LCMS (ESI-MS): Calculation of molecular weight C 21 H 12 F4N2 368.1 m / z, observed value 368.7 [M+H]⁺.

[0396] 1H NMR (400 MHz, CDCl3) δ: 9.00 (d, J = 8.4 Hz, 1H), 7.84–7.82 (m,2H), 7.68 (s, 1H), 7.48–7.40 (m, 2H), 7.28–7.27 (m, 1H), 7.24 (s, 1H), 6.85(s, 1H), 2.53(s, 3H).

[0397] <Preparation Example 9> Preparation Example 21 (the compound represented by chemical formula 23; AMD-D-262)

[0398] The compound of Example 21 was synthesized according to the following reaction formula 14.

[0399] [Reaction Formula 14]

[0400] .

[0401] Step 1. Preparation of 5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-carbonitrile

[0402] The intermediate 5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (200 mg, 0.799 mmol) from step 2 of Example 18 and 2-bromo-1-(thiophen-2-yl)ethane-1-one (196.7 mg, 0.959 mmol) were mixed in acetonitrile (9 mL), and cesium carbonate (520.86 mg, 1.598 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio 10:90) to give 5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrole-3-onitrile (80 mg, yield 24%), which is a yellow solid.

[0403] Step 2. Preparation of 3-methyl-6-(thiophen-2-yl)-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 21)

[0404] A mixture of 5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-2-(4-(trifluoromethyl)phenyl)-1H-pyrrolo-3-onitrile (80 mg, 0.213 mmol), trifluoromethanesulfonic acid (320.72 mg, 2.137 mmol), and 1,2-dichloroethane (4 mL) was stirred at 60 °C for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) to give 3-methyl-6-(thiophen-2-yl)-8-(trifluoromethyl)pyrrolo[2,1-a]isoquinoline-1-onitrile (11 mg, 13% yield) as a white solid.

[0405] LCMS (ESI-MS): Calculation of molecular weight C 19 H 11 F3N2S 356.1 m / z, observed value 356.7 [M+H]⁺.

[0406] 1 H NMR (400 MHz, CDCl3) δ: 8.99 (d, J = 8.8 Hz, 1H), 8.18 (s, 1H), 7.85–7.84 (m, 2H), 7.52–7.51 (m, 1H), 7.27 (s, 1H), 7.24–7.23 (m, 1H), 6.84(s, 1H), 2.53(s, 3H).

[0407] <Preparation Example 10> Preparation Example 22 (Compound represented by chemical formula 24; IK 15g)

[0408] The compound of Example 22 was synthesized according to the following reaction formula 15.

[0409] [Reaction Formula 15]

[0410] .

[0411] Step 1. Preparation of 2-(4-methoxybenzoyl)-4-oxopentanenitrile

[0412] 3-(4-methoxyphenyl)-3-oxopropionitrile (2 g, 0.0114 mol), 1-chloropropane-2-one (1.16 g, 0.0125 mol), and potassium carbonate (3.94 g, 0.0285 mol) were mixed in ethanol (24 mL) and the mixture was stirred at 25 °C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 40:60) to give 2-(4-methoxybenzoyl)-4-oxopentanonitrile (2.3 g, 78% yield) as a white solid.

[0413] Step 2. Preparation of 2-(4-methoxyphenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0414] 2-(4-methoxybenzoyl)-4-oxopentanilonitrile (1.8 g, 0.0078 mol) and ammonium acetate (1.26 g, 0.0164 mol) were mixed in ethanol (15 mL) and stirred at 80 °C for 5 hours. Water (200 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (200 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The concentrate was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 30:70) to give 2-(4-methoxyphenyl)-5-methyl-1H-pyrrole-3-onitrile (1.7 g, 92% yield) as a yellow solid.

[0415] Step 3. Preparation of 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrole-3-carbonitrile

[0416] 2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.9423 mmol), 2-bromo-1-phenylethane-1-one (225.07 mg, 1.1307 mmol), and cesium carbonate (614.04 mg, 1.8846 mmol) were mixed in acetonitrile (5 mL) and stirred at 60 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 30:70) to give 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (140 mg, 40% yield) as a yellow solid.

[0417] Step 4. Preparation of 8-methoxy-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 22)

[0418] A mixture of 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (70 mg, 0.2119 mmol), trifluoromethanesulfonic acid (318.02 mg, 2.119 mmol), and 1,2-dichloroethane (6 mL) was stirred at 25 °C for 1 hour. The reaction was quenched with a saturated aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by preparative high-performance liquid chromatography (Prep-HPLC) (Gemini 5 μm C18, 150 × 21.2 mm, 70%–95% acetonitrile / 0.1% formic acid) to give 8-methoxy-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (AMD-D-263) (23 mg, 33% yield) as a white solid.

[0419] LCMS (ESI-MS): Calculation of molecular weight C 21 H 16 N₂O 312.1 m / z, observed value 312.7 [M+H]⁺.

[0420] 1H NMR (400 MHz, CDCl3) δ: 8.84 (d, J = 9.0 Hz, 1H), 7.59 (s, 1H), 7.56–7.49 (m, 5H), 7.26–7.22 (m, 1H), 7.05 (s, 1H), 6.71 (s, 1H), 3.79 (s,3H), 2.46 (s,3H).

[0421] <Preparation Example 11> Preparation Example 23 (the compound represented by chemical formula 25; AMD-D-264)

[0422] The compound of Example 23 was synthesized according to the following reaction formula 16.

[0423] [Reaction Formula 16]

[0424] .

[0425] Step 1. Preparation of 2-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0426] The intermediate 2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.9423 mmol), 2-bromo-1-(4-methoxyphenyl)ethane-1-one (259.0 mg, 1.1307 mmol), and cesium carbonate (614.04 mg, 1.8846 mmol) from step 2 of Example 22 were mixed in acetonitrile (5 mL) and stirred at 60 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 30:70) to give 2-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (340 mg, 80% yield), which was a yellow solid.

[0427] Step 2. Preparation of 8-methoxy-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 23)

[0428] A mixture of 2-(4-methoxyphenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.2775 mmol), trifluoromethanesulfonic acid (416.47 mg, 2.775 mmol), and 1,2-dichloroethane (6 mL) was stirred at 25 °C for 3 hours. The reaction was quenched with a saturated aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by preparative high performance liquid chromatography (Prep-HPLC) (Gemini 5 μm C18, 150 × 21.2 mm, 70%–95% acetonitrile / 0.1% formic acid) to give 8-methoxy-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (AMD-D-264) (17 mg, yield 17%), which was a white solid.

[0429] LCMS (ESI-MS): Calculation of molecular weight C 22 H 18 N2O2 342.1 m / z, observed value 342.7 [M+H]⁺.

[0430] 1 H NMR (400 MHz, CDCl3) δ: 8.83 (d, J = 9.0 Hz, 1H), 7.57 (s, 1H), 7.43–7.37 (m, 2H), 7.24 (m, 1H), 7.09–7.02 (m, 3H), 6.70 (s, 1H), 3.91 (s, 3H), 3.79 (s, 3H), 2.46 (s, 3H).

[0431] <Preparation Example 12> Preparation Example 24 (the compound represented by chemical formula 26; AMD-D-265)

[0432] The compound of Example 24 was synthesized according to the following reaction formula 17.

[0433] [Reaction Formula 17]

[0434] .

[0435] Step 1. Preparation of 1-(2-(4-fluorophenyl)-2-oxoethyl)-2-(4-methoxyphenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0436] The intermediate 2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.9423 mmol), 2-bromo-1-(4-fluorophenyl)ethane-1-one (245.41 mg, 1.1307 mmol), and cesium carbonate (614.04 mg, 1.8846 mmol) from step 2 of Example 22 were mixed in acetonitrile (15 mL) and stirred at 60 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 30:70) to give 1-(2-(4-fluorophenyl)-2-oxoethyl)-2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, yield 27%), which was a yellow oil.

[0437] Step 2. Preparation of 6-(4-fluorophenyl)-8-methoxy-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 24)

[0438] A mixture of 1-(2-(4-fluorophenyl)-2-oxoethyl)-2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (80 mg, 0.2296 mmol), trifluoromethanesulfonic acid (344.58 mg, 2.296 mmol), and 1,2-dichloroethane (6 mL) was stirred at 25 °C for 2 hours. The reaction was quenched with a saturated aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by preparative high performance liquid chromatography (Prep-HPLC) (Gemini 5 μm C18, 150 × 21.2 mm, 70%–95% acetonitrile / 0.1% formic acid) to give 6-(4-fluorophenyl)-8-methoxy-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (14.3 mg, yield 18%), which was a white solid.

[0439] LCMS (ESI-MS): Calculation of molecular weight C 21 H15 FN2O 330.1 m / z, observed value 330.7 [M+H]⁺.

[0440] 1 H NMR (400 MHz, CDCl3) δ: 8.84 (d, J = 9.0 Hz, 1H), 7.57 (s, 1H), 7.48–7.42 (m, 2H), 7.24–7.18 (m, 3H), 6.97 (s, 1H), 6.71 (s, 1H), 3.80 (s,3H), 2.47 (s,3H).

[0441] <Preparation Example 13> Preparation Example 25 (the compound represented by chemical formula 27; AMD-D-267)

[0442] The compound of Example 25 was synthesized according to the following reaction formula 18.

[0443] [Reaction Formula 18]

[0444] .

[0445] Step 1. Preparation of 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-carbonitrile

[0446] The intermediate 2-(4-methoxyphenyl)-5-methyl-1H-pyrrolo-3-onitrile (350 mg, 1.649 mmol), 2-bromo-1-(thiophen-2-yl)ethane-1-one (405.8 mg, 1.979 mmol), and cesium carbonate (1074.5 mg, 3.298 mmol) from step 2 of Example 22 were mixed in acetonitrile (15 mL) and stirred at 60 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 30:70) to give 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrolo-3-onitrile (100 mg, yield 16%), which was a yellow oil.

[0447] Step 2. Preparation of 8-methoxy-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 25)

[0448] 2-(4-methoxyphenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrolo-3-onitrile (90 mg, 0.2675 mmol), trifluoromethanesulfonic acid (401.5 mg, 2.675 mmol), and 1,2-dichloroethane (6 mL) were mixed and stirred at 25 °C for 3 hours. The reaction was quenched with a saturated aqueous sodium bicarbonate solution and extracted with dichloroethane. The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by preparative high performance liquid chromatography (Prep-HPLC) (Gemini 5 μm C18, 150 × 21.2 mm, 60%–95% acetonitrile / 0.1% formic acid) to give 8-methoxy-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-onitrile (15 mg, yield 17%), which was a white solid.

[0449] LCMS (ESI-MS): Calculation of molecular weight C 19 H 14 N2OS 318.1 m / z, observed 319.1 [M+H]⁺.

[0450] 1 H NMR (400 MHz, CDCl3) δ: 8.82 (d, J = 9.0 Hz, 1H), 7.74 (s, 1H), 7.49–7.47 (m, 1H), 7.35 (s, 1H), 7.27 (s, 1H), 7.25–7.21 (m, 2H), 6.70 (s,1H), 3.84 (s, 3H), 2.47 (s, 3H).

[0451] <Preparation Example 14> Preparation Example 26 (the compound represented by chemical formula 28; IK15j)

[0452] The compound of Example 26 was synthesized according to the following reaction formula 19.

[0453] [Reaction Formula 19]

[0454] .

[0455] Step 1. Preparation of 2-(4-methylbenzoyl)-4-oxopentanenitrile

[0456] 3-Oxo-3-(p-Tolyl)propionitrile (5 g, 31.44 mmol) and 1-chloropropane-2-one (4.3 g, 47.17 mmol) were mixed in ethanol (80 mL) and potassium carbonate (8.7 g, 62.88 mmol) was added. The mixture was stirred at 25 °C for 2 hours under a nitrogen atmosphere. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (120 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) to give 2-(4-methylbenzoyl)-4-oxopentanonitrile (3.1 g, 49% yield).

[0457] Step 2. Preparation of 5-methyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0458] 2-(4-methylbenzoyl)-4-oxopentanilonitrile (3.1 g, 14.42 mmol) and ammonium acetate (2.2 g, 28.83 mmol) were mixed in ethanol (50 mL) and stirred at 80 °C for 5 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 5-methyl-2-(p-tolyl)-1H-pyrrole-3-onitrile (2.0 g, 71% yield) as a yellow solid.

[0459] Step 3. Preparation of 5-methyl-1-(2-oxo-2-phenylethyl)-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0460] 5-Methyl-2-(p-Tolyl)-1H-pyrrolo-3-onitrile (1.0 g, 5.1 mmol) and 2-bromo-1-phenylethane-1-one (2.0 g, 10.2 mmol) were mixed in acetonitrile (60 mL), and cesium carbonate (3.3 g, 10.2 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (40 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (80 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 5-methyl-1-(2-oxo-2-phenylethyl)-2-(p-Tolyl)-1H-pyrrolo-3-onitrile (0.5 g, 31% yield) as a yellow solid.

[0461] Step 4. Preparation of 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 26)

[0462] A mixture of 5-methyl-1-(2-oxo-2-phenylethyl)-2-(p-tolyl)-1H-pyrrolo-3-onitrile (500 mg, 1.6 mmol), trifluoromethanesulfonic acid (2.4 g, 16.0 mmol), and 1,2-dichloroethane (30 mL) was stirred at room temperature for 2 hours. Water (25 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 8:92) to give 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (AMD-D-268) (300 mg, 63% yield) as a white solid.

[0463] LCMS (ESI-MS): Calculation of molecular weight C 21 H 16 N2 296.1 m / z, observed value 296.7 [M+H]⁺.

[0464] 1 H NMR (400 MHz, CDCl3) δ: 8.80 (d, J = 8.4 Hz, 1H), 7.66–7.35 (m,8H), 6.73 (s, 1H), 2.47 (s, 3H), 2.42 (s, 3H).

[0465] <Preparation Example 15> Preparation Example 27 (the compound represented by chemical formula 29; IK15k)

[0466] The compound of Example 27 was synthesized according to the following reaction formula 20.

[0467] [Reaction 20]

[0468] .

[0469] Step 1. Preparation of 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile)

[0470] The intermediate 5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (250 mg, 1.28 mmol) from step 2 of Example 26 and 2-bromo-1-(4-methoxyphenyl)ethane-1-one (440 mg, 1.92 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (832 mg, 2.56 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-onitrile (120 mg, yield 27%), which was a yellow solid.

[0471] Step 2. Preparation of 6-(4-methoxyphenyl)-3,8-dimethylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 27)

[0472] 1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (50 mg, 0.15 mmol), trifluoromethanesulfonic acid (225 mg, 1.5 mmol), and 1,2-dichloroethane (2 mL) were mixed and stirred at room temperature for 2 hours. Water (4 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 8:92) to give 6-(4-methoxyphenyl)-3,8-dimethylpyrrolo[2,1-a]isoquinoline-1-onitrile (10.7 mg, 22% yield) as a white solid.

[0473] LCMS (ESI-MS): Calculation of molecular weight C 22 H 18 N2O 326.1 m / z, observed value 326.8 [M+H]⁺.

[0474] 1 H NMR (400 MHz, CDCl3) δ: 8.79 (d, J = 8.4 Hz, 1H), 7.56 (s, 1H), 7.48–7.33 (m, 4H), 7.09–7.01 (m, 2H), 6.72 (s, 1H), 3.91 (s, 3H), 3.00–2.25(m, 6H).

[0475] <Preparation Example 16> Preparation Example 28 (the compound represented by chemical formula 30; AMD-D-270)

[0476] The compound of Example 28 was synthesized according to the following reaction formula 21.

[0477] [Reaction 21]

[0478] .

[0479] Step 1. Preparation of 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0480] The intermediate 5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (250 mg, 1.28 mmol) from step 2 of Example 26 and 2-bromo-1-(4-fluorophenyl)ethane-1-one (416 mg, 1.92 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (832 mg, 2.56 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (110 mg, 26% yield) as a yellow solid.

[0481] Step 2. Preparation of 6-(4-fluorophenyl)-3,8-dimethylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 28)

[0482] 1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (50 mg, 0.15 mmol), trifluoromethanesulfonic acid (225 mg, 1.5 mmol), and 1,2-dichloroethane (2 mL) were mixed and stirred at room temperature for 2 hours. Water (4 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 9:91) to give 6-(4-fluorophenyl)-3,8-dimethylpyrrolo[2,1-a]isoquinoline-1-onitrile (10.9 mg, 23% yield) as a white solid.

[0483] LCMS (ESI-MS): Calculation of molecular weight C 21 H 15 FN2 314.1 m / z, observed value 314.7 [M+H]⁺.

[0484] 1H NMR (400 MHz, CDCl3) δ: 8.80 (d, J = 8.4 Hz, 1H), 7.56 (s, 1H), 7.50–7.38 (m, 3H), 7.33 (s, 1H), 7.25–7.14 (m, 2H), 6.74 (s, 1H), 2.48–2.43(m, 6H).

[0485] <Preparation Example 17> Preparation Example 29 (the compound represented by chemical formula 31; AMD-D-272)

[0486] The compound of Example 29 was synthesized according to the following reaction formula 22.

[0487] [Reaction 22]

[0488] .

[0489] Step 1. Preparation of 5-methyl-2-(4-methylcyclohexa-1,3-dien-1-yl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-carbonitrile

[0490] The intermediate 5-methyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (250 mg, 1.28 mmol) from step 2 of Example 26 and 2-bromo-1-(thiophene-2-yl)ethane-1-one (394 mg, 1.92 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (832 mg, 2.56 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 20:80) to give 5-methyl-2-(4-methylcyclohexyl-1,3-dien-1-yl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-onitrile (100 mg, yield 24%), which was a yellow solid.

[0491] Step 2. Preparation of 3,8-dimethyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 29)

[0492] A mixture of 5-methyl-2-(4-methylcyclohexyl-1,3-dien-1-yl)-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrolo-3-onitrile (50 mg, 0.16 mmol), trifluoromethanesulfonic acid (240 mg, 1.6 mmol), and 1,2-dichloroethane (2 mL) was stirred at room temperature for 2 hours. Water (4 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 9:91) to give 3,8-dimethyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-onitrile (10.4 mg, 22% yield) as a white solid.

[0493] LCMS (ESI-MS): Calculation of molecular weight C 19 H 14 N2S 302.1 m / z, observed value 302.6 [M+H]⁺.

[0494] 1 H NMR (400 MHz, CDCl3) δ: 8.78 (d, J = 8.4 Hz, 1H), 7.73–7.67 (m,2H), 7.53–7.42 (m, 2H), 7.27–7.11 (m, 2H), 6.73 (s, 1H), 2.63–2.22 (m, 6H).

[0495] <Preparation Example 18> Preparation Example 30 (the compound represented by chemical formula 32; AMD-D-273)

[0496] The compound of Example 30 was synthesized according to the following reaction formula 23.

[0497] [Reaction 23]

[0498] .

[0499] Step 1. Preparation of 2-(3-fluorobenzoyl)-4-oxopentanenitrile

[0500] 1-Chloropropane-2-one (2.19 g, 23.65 mmol) was added to 3-(3-fluorophenyl)-3-oxopropionitrile (3.5 g, 21.5 mmol), potassium carbonate (7.42 g, 23.65 mmol), and ethanol (50 mL). The mixture was stirred at 25 °C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (60 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(3-fluorobenzoyl)-4-oxopentanilonitrile (4.6 g, 97% yield), which was a yellow oil.

[0501] Step 2. Preparation of 2-(3-fluorophenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0502] 2-(3-fluorobenzoyl)-4-oxopentanilonitrile (4.7 g, 21.4 mmol), ammonium acetate (3.3 g, 42.8 mmol), and ethanol (50 mL) were mixed and stirred at 80 °C for 5 hours. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(3-fluorophenyl)-5-methyl-1H-pyrrole-3-onitrile (2.84 g, 66% yield) as a yellow solid.

[0503] Step 3. Preparation of 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrole-3-carbonitrile

[0504] 2-(3-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (300 mg, 1.5 mmol), 2-bromo-1-phenylethane-1-one (450 mg, 2.25 mmol), and cesium carbonate (975 mg, 3.0 mmol) were mixed in acetonitrile (17 mL) and stirred at 60 °C for 24 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (40 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (120 mg, 38% yield) as a yellow solid.

[0505] Step 4. Preparation of 9-fluoro-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 30)

[0506] 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (60 mg, 0.19 mmol), trifluoromethanesulfonic acid (285 mg, 1.9 mmol), and 1,2-dichloroethane (3 mL) were mixed and stirred at room temperature for 2 hours. Water (4 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 9-fluoro-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (10.9 mg, 19% yield) as a white solid.

[0507] LCMS (ESI-MS): Calculation of molecular weight C 20 H 13 FN2 300.1 m / z, observed value 300.7 [M+H]⁺.

[0508] 1 H NMR (400 MHz, CDCl3) δ: 8.55 (dd, J = 7.2, 2.4 Hz, 1H), 7.73–7.38(m, 7H), 7.24–7.11 (m, 1H), 6.78 (s, 1H), 2.49 (s, 3H).

[0509] <Preparation Example 19> Preparation Example 31 (the compound represented by chemical formula 33; AMD-D-274)

[0510] The compound of Example 31 was synthesized according to the following reaction formula 24.

[0511] [Reaction 24]

[0512] .

[0513] Step 1. Preparation of 2-(3-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0514] The intermediate 2-(3-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (270 mg, 1.35 mmol) from step 2 of Example 30 and 2-bromo-1-(4-methoxyphenyl)ethane-1-one (462 mg, 2.03 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (877 mg, 2.7 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (40 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio 16:84) to give 2-(3-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (150 mg, yield 43%), which is a yellow solid.

[0515] Step 2. Preparation of 9-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 31)

[0516] 2-(3-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.29 mmol), trifluoromethanesulfonic acid (435 mg, 2.9 mmol), and 1,2-dichloroethane (5 mL) were mixed and stirred at room temperature for 2 hours. Water (5 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 12:88) to give 9-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (AMD-D-274) (10.6 mg, 11% yield) as a white solid.

[0517] LCMS (ESI-MS): Calculation of molecular weight C 21 H 15 FN2O 330.1 m / z, observed value 330.7 [M+H]⁺.

[0518] 1 H NMR (400 MHz, CDCl3) δ: 8.53 (dd, J = 9.6, 2.4 Hz, 1H), 7.65–7.62(m, 1H), 7.56 (s, 1H), 7.45–7.34 (m, 2H), 7.24–7.13 (m, 1H), 7.10–7.02 (m,2H), 6.77 (s, 1H), 3.91 (s, 3H), 2.49 (s, 3H).

[0519] <Preparation Example 20> Preparation Example 32 (the compound represented by chemical formula 34; AMD-D-275)

[0520] The compound of Example 32 was synthesized according to the following reaction formula 25.

[0521] [Reaction 25]

[0522] .

[0523] Step 1. Preparation of 2-(3-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0524] The intermediate 2-(3-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (270 mg, 1.35 mmol) from step 2 of Example 30 and 2-bromo-1-(4-fluorophenyl)ethane-1-one (440 mg, 2.03 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (877 mg, 2.7 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (40 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio 16:84) to give 2-(3-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (100 mg, yield 22%), which is a yellow solid.

[0525] Step 2. Preparation of 9-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 32)

[0526] 2-(3-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.30 mmol), trifluoromethanesulfonic acid (450 mg, 3.0 mmol), and 1,2-dichloroethane (5 mL) were mixed and stirred at room temperature for 2 hours. Water (5 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 13:87) to give 9-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (10.6 mg, 11% yield) as a white solid.

[0527] LCMS (ESI-MS): Calculation of molecular weight C 20 H 12 F2N2 318.1 m / z, observed value 318.7 [M+H]⁺.

[0528] 1H NMR (400 MHz, CDCl3) δ: 8.54 (dd, J = 9.6, 2.4 Hz, 1H), 7.58–7.54(m, 2H), 7.49–7.42 (m, 2H), 7.28–7.10 (m, 3H), 6.78 (s, 1H), 2.50 (s, 3H).

[0529] <Preparation Example 21> Preparation Example 33 (the compound represented by chemical formula 35; AMD-D-277)

[0530] The compound of Example 33 was synthesized according to the following reaction formula 26.

[0531] [Reaction 26]

[0532] .

[0533] Step 1. Preparation of 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-carbonitrile

[0534] The intermediate 2-(3-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (270 mg, 1.35 mmol) from step 2 of Example 30 and 2-bromo-1-(thiophen-2-yl)ethane-1-one (416 mg, 2.03 mmol) were mixed in acetonitrile (15 mL), and cesium carbonate (877 mg, 2.7 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (40 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 16:84) to give 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrole-3-onitrile (150 mg, yield 46%), which was a yellow solid.

[0535] Step 2. Preparation of 9-fluoro-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitril (Example 33)

[0536] 2-(3-fluorophenyl)-5-methyl-1-(2-oxo-2-(thiophen-2-yl)ethyl)-1H-pyrrolo-3-onitrile (80 mg, 0.25 mmol), trifluoromethanesulfonic acid (375 mg, 2.5 mmol), and 1,2-dichloroethane (3 mL) were mixed and stirred at room temperature for 2 hours. Water (5 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 20:80) to give 9-fluoro-3-methyl-6-(thiophen-2-yl)pyrrolo[2,1-a]isoquinoline-1-onitrile (10.9 mg, 14% yield) as a white solid.

[0537] LCMS (ESI-MS): Calculation of molecular weight C 18 H 11 FN2S 306.1 m / z, observed value 306.7 [M+H]⁺.

[0538] 1 H NMR (400 MHz, CDCl3) δ: 8.52 (dd, J = 9.6, 2.4 Hz, 1H), 7.92–7.88(m, 1H), 7.73 (s, 1H), 7.49 (d, J = 4.8 Hz, 1H), 7.24–7.21 (m, 3H), 6.78 (s,1H), 2.50 (s,3H).

[0539] <Preparation Example 22> Preparation Example 34 (the compound represented by chemical formula 36; AMD-D-278)

[0540] The compound of Example 34 was synthesized according to the following reaction formula 27.

[0541] [Reaction 27]

[0542] .

[0543] Step 1. Preparation of 2-(2-fluorobenzoyl)-4-oxopentanenitrile

[0544] 1-Chloropropane-2-one (2.02 g, 21.78 mmol) was added to 3-(2-fluorophenyl)-3-oxopropionitrile (3.23 g, 19.8 mmol), potassium carbonate (6.83 g, 49.5 mmol), and ethanol (50 mL). The mixture was stirred at 25 °C for 2 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(2-fluorobenzoyl)-4-oxopentanonitrile (4.3 g, 98% yield) as a yellow oil.

[0545] Step 2. Preparation of 2-(2-fluorophenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0546] 2-(2-fluorobenzoyl)-4-oxopentanilonitrile (4.66 g, 21.3 mmol), ammonium acetate (3.28 g, 42.6 mmol), and ethanol (50 mL) were mixed and stirred at 80 °C for 5 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 2-(2-fluorophenyl)-5-methyl-1H-pyrrole-3-onitrile (3.04 g, 71% yield) as a yellow solid.

[0547] Step 3. Preparation of 2-(2-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrole-3-carbonitrile

[0548] 2-(2-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.999 mmol) and 2-bromo-1-phenylethane-1-one (238.6 mg, 1.20 mmol) were dissolved in acetonitrile (10 mL), and cesium carbonate (651 mg, 1.998 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (30 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (30 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 2-(2-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (230 mg, 72% yield) as a yellow oil.

[0549] Step 4. Preparation of 10-fluoro-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 34)

[0550] 2-(2-fluorophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (100 mg, 0.314 mmol) was dissolved in 1,2-dichloroethane (8 mL), and then trifluoromethanesulfonic acid (235.7 mg, 1.57 mmol) was added. The mixture was stirred at 60 °C for 1 hour. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 8:92) to give 10-fluoro-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (10.6 mg, 11% yield) as a white solid.

[0551] LCMS (ESI-MS): Calculation of molecular weight C 20 H 13 FN2 300.1 m / z, observed value 301.1 [M+H]⁺.

[0552] 1H NMR (400 MHz, CDCl3) δ: 7.64 (s, 1H), 7.56–7.46 (m, 5H), 7.43–7.39 (m, 2H), 7.38–7.33 (m, 1H), 6.90 (s, 1H), 2.50 (s, 3H).

[0553] <Preparation Example 23> Preparation Example 35 (the compound represented by chemical formula 37; AMD-D-279)

[0554] The compound of Example 35 was synthesized according to the following reaction formula 28.

[0555] [Reaction 28]

[0556] .

[0557] Step 1. Preparation of 2-(2-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0558] The intermediates 2-(2-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.999 mmol) and 2-bromo-1-(4-methoxyphenyl)ethane-1-one (343.23 mg, 1.50 mmol) from step 2 of Example 34 were dissolved in acetonitrile (10 mL), and cesium carbonate (651 mg, 1.998 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (30 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (30 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, volume ratio 15:85) to give 2-(2-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (330 mg, yield 94%), which was a yellow oil.

[0559] Step 2. Preparation of 10-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 35)

[0560] 2-(2-fluorophenyl)-1-(2-(4-methoxyphenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (800 mg, 2.299 mmol), trifluoromethanesulfonic acid (3.448 g, 22.99 mmol), and 1,2-dichloroethane (50 mL) were mixed and stirred at 80 °C for 1 hour. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (70 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 10-fluoro-6-(4-methoxyphenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (22 mg, 1% yield) as a white solid.

[0561] LCMS (ESI-MS): Calculation of molecular weight C 21 H 15 FN2O 330.1 m / z, observed value 330.7 [M+H]⁺.

[0562] 1 H NMR (400 MHz, DMSO-d6) δ: 8.02 (s, 1H), 7.63–7.55 (m, 2H), 7.51–7.44 (m, 3H), 7.16–7.06 (m, 3H), 3.85 (s, 3H), 2.54 (s, 3H).

[0563] <Preparation Example 24> Preparation Example 36 (the compound represented by chemical formula 38; AMD-D-280)

[0564] The compound of Example 36 was synthesized according to the following reaction formula 29.

[0565] [Reaction 29]

[0566] .

[0567] Step 1. Preparation of 2-(2-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0568] The intermediates 2-(2-fluorophenyl)-5-methyl-1H-pyrrolo-3-onitrile (200 mg, 0.999 mmol) and 2-bromo-1-(4-fluorophenyl)ethane-1-one (325.19 mg, 1.50 mmol) from step 2 of Example 34 were dissolved in acetonitrile (10 mL), and cesium carbonate (651 mg, 1.998 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (30 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (30 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 2-(2-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrole-3-onitrile (230 mg, yield 68%), which was a yellow oil.

[0569] Step 2. Preparation of 10-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 36)

[0570] 2-(2-fluorophenyl)-1-(2-(4-fluorophenyl)-2-oxoethyl)-5-methyl-1H-pyrrolo-3-onitrile (100 mg, 0.297 mmol), trifluoromethanesulfonic acid (446.19 mg, 2.973 mmol), and 1,2-dichloroethane (5 mL) were mixed and stirred at 70 °C for 1 hour. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 10-fluoro-6-(4-fluorophenyl)-3-methylpyrrolo[2,1-a]isoquinoline-1-onitrile (15 mg, 15% yield) as a white solid.

[0571] LCMS (ESI-MS): Calculation of molecular weight C 20 H 12 F2N2 318.1 m / z, observed value 318.7 [M+H]⁺.

[0572] 1H NMR (400 MHz, DMSO-d6) δ: 8.08 (s, 1H), 7.62–7.57 (m, 4H), 7.46–7.35 (m, 3H), 7.08 (s, 1H), 2.54 (s, 3H).

[0573] <Preparation Example 25> Preparation Example 37 (the compound represented by chemical formula 39; AMD-D-284)

[0574] The compound of Example 37 was synthesized according to the following reaction formula 30.

[0575] [Reaction 30]

[0576] .

[0577] Step 1. Preparation of 2-(4-methylbenzoyl)-4-oxo-4-phenylbutanenitrile

[0578] 3-Oxo-3-(p-Tolyl)propionitrile (500 mg, 3.14 mmol) and 2-chloro-1-phenylethane-1-one (534 mg, 3.45 mmol) were mixed in ethanol (8 mL), and potassium carbonate (1085 mg, 7.85 mmol) was added under a nitrogen atmosphere. The mixture was stirred at 25 °C for 2 hours. Water (5 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (15 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) to give 2-(4-methylbenzoyl)-4-oxo-4-phenylbutyronitrile (260 mg, 29% yield).

[0579] Step 2. Preparation of 5-phenyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0580] 2-(4-methylbenzoyl)-4-oxo-4-phenylbutyronitrile (260 mg, 0.94 mmol), ammonium acetate (145 mg, 1.88 mmol), and ethanol (5 mL) were mixed and stirred at 80 °C for 5 hours. Water (5 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 10:90) to give 5-phenyl-2-(p-tolyl)-1H-pyrrole-3-onitrile (166 mg, 66% yield) as a yellow solid.

[0581] Step 3. Preparation of 1-(2-oxo-2-phenylethyl)-5-phenyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0582] 5-Phenylacetyl-2-(p-Tolyl)-1H-pyrrolo-3-onitrile (160 mg, 0.62 mmol) and 2-bromo-1-phenylethane-1-one (185 mg, 0.93 mmol) were dissolved in acetonitrile (6 mL), and cesium carbonate (404 mg, 1.24 mmol) was added. The mixture was stirred at 60 °C for 24 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) to give 1-(2-oxo-2-phenylethyl)-5-phenyl-2-(p-Tolyl)-1H-pyrrolo-3-onitrile (160 mg, 68% yield) as a yellow solid.

[0583] Step 4. Preparation of 8-methyl-3,6-diphenylpyrrolo[2,1-a] isoquinoline-1-carbonitrile (Example 37)

[0584] A mixture of 1-(2-oxo-2-phenylethyl)-5-phenyl-2-(p-tolyl)-1H-pyrrolo-3-onitrile (100 mg, 0.26 mmol), trifluoromethanesulfonic acid (199 mg, 1.33 mmol), and 1,2-dichloroethane (10 mL) was stirred at 80 °C for 0.5 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated. The residue was purified by preparative high-performance liquid chromatography (Prep-HPLC) (20%–60% acetonitrile / 0.1% ammonia) to give 8-methyl-3,6-diphenylpyrrolo[2,1-a]isoquinoline-1-onitrile (23.3 mg, 24% yield) as a white solid.

[0585] LCMS (ESI-MS): Calculation of molecular weight C 26 H 18 N2 358.1 m / z, observed value 359.0 [M+H]⁺.

[0586] 1 H NMR (400 MHz, CDCl3) δ: 8.11 (d, J = 7.6 Hz, 1H), 7.79 (s, 1H), 7.57–7.50 (m, 4H), 7.47–7.45 (m, 2H), 7.45–7.40 (m, 3H), 7.39–7.33 (m, 3H), 7.29 (s, 1H), 2.43 (s, 3H).

[0587] <Preparation Example 26> Preparation Example 38 (the compound represented by chemical formula 40; AMD-D-286)

[0588] The compound of Example 38 was synthesized according to the following reaction formula 31.

[0589] [Reaction Formula 31]

[0590] .

[0591] Step 1. Preparation of 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carboxamide (Example 38)

[0592] 3,8-Dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-nitrile (Example 26, 300 mg, 1.01 mmol) and sulfuric acid (3 mL) were mixed and stirred at 25 °C for 16 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). After washing, the mixture was dried over sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (methanol / dichloromethane, v / v 10:90) to give 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carboxamide (150 mg, 47% yield) as a white solid.

[0593] LCMS (ESI-MS): Calculation of molecular weight C 21 H 18 N 2O 314.1 m / z, observed value 314.8 [M+H]⁺.

[0594] 1 H NMR (400 MHz, DMSO-d6) δ: 9.59 (d, J = 8.4 Hz, 1H), 7.80 (s, 1H), 7.62–7.45 (m, 5H), 7.35 (d, J = 8.4 Hz, 1H), 7.28 (s, 1H), 6.90 (s, 1H), 2.47(s, 3H), 2.34(s, 3H)

[0595] <Preparation Example 27> Preparation Example 39 (the compound represented by chemical formula 41; AMD-D-287)

[0596] The compound of Example 39 was synthesized according to the following reaction formula 32.

[0597] [Reaction 32]

[0598] .

[0599] Step 1. Preparation of ethyl 2-(4-methylbenzoyl)-4-oxopentanoate

[0600] Ethyl 3-(4-methylphenyl)-3-oxopropionate (7 g, 33.9 mmol), sodium iodide (1.02 g, 6.70 mmol), and potassium carbonate (11.71 g, 84.7 mmol) were mixed in acetone (50 mL), and 1-chloropropane-2-one (3.45 g, 37.29 mmol) was added. The mixture was stirred at 60 °C for 5 hours. The reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was purified by silica gel column chromatography (ethyl acetate / hexane, v / v 5:95) to give ethyl 2-(4-methylbenzoyl)-4-oxopentanoate (8 g, 90% yield), which was a yellow solid.

[0601] Step 2. Preparation of ethyl 5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylate

[0602] Ethyl 2-(4-methylbenzoyl)-4-oxovalerate (8 g, 30.5 mmol) was dissolved in ethanol (80 mL), and then ammonium acetate (4.70 g, 61.0 mmol) was added. The mixture was stirred at 80 °C for 5 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (ethyl acetate / hexane, v / v 10:90) to give ethyl 5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylic acid (7 g, 94% yield) as a yellow solid.

[0603] Step 3. Preparation of ethyl 1-(2-hydroxy-2-phenylethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylate

[0604] Ethyl 5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylate (2 g, 8 mmol), 2-phenylethylene oxide (1.48 g, 12 mmol), and potassium tert-butoxide (2.76 g, 24 mmol) were mixed in dimethylformamide (DMF, 60 mL) and heated at 80 °C for 2 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated. Crystallization, filtration, and vacuum drying with an ethyl acetate / hexane (1:1) mixture gave ethyl 1-(2-hydroxy-2-phenylethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylate (500 mg, 17% yield), as a white solid.

[0605] Step 4. Preparation of ethyl 5-methyl-1-(2-oxo-2-phenylethyl)-2-(p-tolyl)-1H-pyrrole-3-carboxylate

[0606] Ethyl 1-(2-hydroxy-2-phenylethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carboxylic acid (500 mg, 1.375 mmol) and iodobenzoic acid (577.84 mg, 2.063 mmol) were dissolved in ethyl acetate (10 mL) and stirred at 70 °C for 16 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated. Purification by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 15:85) yielded ethyl 5-methyl-1-(2-oxo-2-phenylethyl)-2-(p-tolyl)-1H-pyrrole-3-carboxylic acid (300 mg, 60% yield) as a white solid.

[0607] Step 5. Preparation of ethyl 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carboxylate (Example 39)

[0608] Ethyl 5-methyl-2-(4-methylphenyl)-1-(2-oxo-2-phenylethyl)-pyrrolo-3-carboxylate (100 mg, 0.276 mmol), trifluoromethanesulfonic acid (415.27 mg, 2.767 mmol), and 1,2-dichloroethane (5 mL) were mixed and stirred at 60 °C for 1 hour. The reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (15 mL × 3). The organic layer was collected, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated. Purification by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 5:95) yielded ethyl 3,8-dimethyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carboxylate (7 mg, 7% yield) as a white solid.

[0609] LCMS (ESI-MS): Calculated value C 23 H 21 NO2 343.2 m / z, measured value 343.8 [M+H]⁺.

[0610] 1 H NMR (400 MHz, CDCl3) δ: 9.85 (d, J = 8.4 Hz, 1H), 7.58 (s, 1H), 7.56–7.47 (m, 5H), 7.46–7.34 (m, 2H), 7.06–7.05 (m, 1H), 4.40 (q, J = 7.2 Hz, 2H), 2.47 (s, 3H), 2.40 (s, 3H), 1.44 (t, J = 7.2 Hz, 3H).

[0611] <Preparation Example 28> Preparation Example 40 (the compound represented by chemical formula 42; AMD-D-289)

[0612] The compound of Example 40 was synthesized according to the following reaction formula 33.

[0613] [Reaction 33]

[0614] .

[0615] Step 1. Preparation of 2-(2-(2-fluoroethoxy)ethoxy)ethyl 4-methylbenzenesulfonate

[0616] 2-(2-(2-fluoroethoxy)ethoxy)ethanol (300 mg, 1.97 mmol) and an aqueous solution of sodium hydroxide (5 N, 1 mL) were mixed in tetrahydrofuran (10 mL), followed by the addition of 4-methylbenzenesulfonyl chloride (570 mg, 3.0 mmol). The mixture was stirred at 0 °C for 6 hours. The reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (10 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 20:80) to give the target compound as a yellow oil (220 mg, 36% yield).

[0617] Step 2. Preparation of 2-(4-bromobenzoyl)-4-oxopentanenitrile

[0618] 3-(4-bromophenyl)-3-oxopropionitrile (6.0 g, 26.8 mmol), 1-chloropropane-2-one (4.9 g, 53.6 mmol), and potassium carbonate (7.4 g, 53.6 mmol) were mixed in ethanol (100 mL) and stirred at room temperature (25 °C) for 1 hour. The reaction mixture was diluted with water (100 mL) and extracted with dichloromethane (80 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 13:87) to give a yellow solid (4.1 g, 55% yield).

[0619] Step 3. Preparation of 2-(4-bromophenyl)-5-methyl-1H-pyrrole-3-carbonitrile

[0620] 2-(4-bromobenzoyl)-4-oxopentanilonitrile (4.1 g, 14.6 mmol) and ammonium acetate (2.26 g, 29.3 mmol) were mixed in ethanol (60 mL) and stirred at 80 °C for 5 hours. The reaction mixture was diluted with water (80 mL) and extracted with dichloromethane (60 mL × 3). The organic layer was washed, dried, concentrated, and purified by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 20:80) to give a yellow solid (3.0 g, 79% yield).

[0621] Step 4. Preparation of 2-(4-bromophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrole-3-carbonitrile

[0622] 2-(4-bromophenyl)-5-methyl-1H-pyrrolo-3-onitrile (3.0 g, 11.5 mmol), 2-bromo-1-phenylethane-1-one (4.6 g, 23.0 mmol), and cesium carbonate (7.5 g, 23.0 mmol) were mixed in acetonitrile (150 mL) and stirred at 60 °C for 16 hours. The reaction mixture was diluted with water (200 mL) and extracted with dichloromethane (150 mL × 3). Purification by silica gel column chromatography (ethyl acetate / petroleum ether, v / v 40:60) gave a yellow solid (2.0 g, 46% yield).

[0623] Step 5. Preparation of 8-bromo-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile

[0624] 2-(4-bromophenyl)-5-methyl-1-(2-oxo-2-phenylethyl)-1H-pyrrolo-3-onitrile (2.0 g, 7.9 mmol) and trifluoromethanesulfonic acid (11.9 g, 79.0 mmol) were dissolved in 1,2-dichloroethane (150 mL) and stirred at 60 °C for 6 hours. The reaction mixture was diluted with water (100 mL) and extracted with dichloromethane (80 mL × 3). Purification by silica gel column chromatography (methanol / dichloromethane, v / v 2:98) gave a yellow oil (1.2 g, 42% yield).

[0625] Step 6. Preparation of 8-(hydroxymethyl)-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile

[0626] 8-Bromo-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (300 mg, 0.83 mmol), (tributyltin)methanol (533 mg, 1.66 mmol), and tetrakis(triphenylphosphine)palladium (92 mg, 0.08 mmol) were dissolved in dioxane (10 mL) and stirred at 80 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was diluted with water (20 mL) and extracted with dichloromethane (20 mL × 3) to obtain a red solid (170 mg, 66% yield).

[0627] Step 7. Preparation of 8-((2-(2-(2-fluoroethoxy)ethoxy)ethoxy)methyl)-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 40)

[0628] 8-(hydroxymethyl)-3-methyl-6-phenylpyrrolo[2,1-a]isoquinoline-1-onitrile (60 mg, 0.19 mmol) and sodium hydride (38 mg, 0.95 mmol) were mixed in N,N-dimethylformamide (DMF, 5 mL), followed by the addition of 2-(2-(2-fluoroethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (116 mg, 0.38 mmol). The mixture was stirred at 0 °C for 2 hours, diluted with water (10 mL), and extracted with ethyl acetate (10 mL × 3). Purification by preparative high-performance liquid chromatography (prep-HPLC) (Gemini 5 μm C18 column, 150 × 21.2 mm, acetonitrile / purified water containing 0.1% formic acid 30%–90%) yielded a yellow solid (10.3 mg, 12% yield).

[0629] LCMS (ESI-MS): Calculated value C 27 H 27 FN2O3 446.2 m / z, measured value 446.8 [M+H]⁺.

[0630] 1H NMR (400 MHz, CDCl3): δ 8.89 (d, J = 8.4 Hz, 1H), 7.75–7.43 (m,8H), 6.76 (s, 1H), 4.70–4.55 (m, 3H), 4.53–4.44 (m, 1H), 3.82–3.74 (m, 1H), 3.74–3.54 (m, 9H), 2.49 (s, 3H).

[0631] <Preparation Example 29> Preparation Example 41 (the compound represented by chemical formula 43; AMD-D-271)

[0632] The compound of Example 41 was synthesized according to the following reaction formula 34.

[0633] [Reaction Formula 34]

[0634] .

[0635] Step 1. Preparation of 2-bromo-1-(pyridin-3-yl)ethan-1-ol

[0636] Sodium borohydride (1.6 g, 42 mmol) was suspended in methanol (90 mL) with stirring. Then, a solution of 2-bromo-1-(pyridin-3-yl)ethane-1-one hydrobromide (3.0 g, 10.5 mmol) suspended in methanol (60 mL) was slowly added at -60 °C for 1 hour. After the addition was complete, the reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (100 mL × 3). The organic layer was concentrated under reduced pressure to give 600 mg (28% yield) of 2-bromo-1-(pyridin-3-yl)ethane-1-ol. Since this compound is unstable at 25 °C, it was used directly in the next step without purification.

[0637] Step 2. Preparation of 1-(2-hydroxy-2-(pyridin-3-yl)ethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0638] 5-Methyl-2-(p-Tolyl)-1H-pyrrolo-3-onitrile (400 mg, 2.04 mmol) and potassium tert-butoxy (120 mg, 4.08 mmol) were dissolved in ethanol (10 mL), and then 2-bromo-1-(pyridin-3-yl)ethane-1-ol (494 mg, 2.45 mmol) prepared in step 1 was added. The reaction mixture was stirred at 80 °C for 2 hours. After the reaction was complete, the reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1:1, v / v) to give 160 mg (25% yield) of the target compound 1-(2-hydroxy-2-(pyridin-3-yl)ethyl)-5-methyl-2-(p-tolyl)-1H-pyrrole-3-onitrile in yellow oil form.

[0639] Step 3. Preparation of 5-methyl-1-(2-oxo-2-(pyridin-3-yl)ethyl)-2-(p-tolyl)-1H-pyrrole-3-carbonitrile

[0640] The compound prepared in step 2 (140 mg, 0.38 mmol) and Dess-Martin oxidant (240 mg, 0.57 mmol) were dissolved in dichloromethane (5 mL) and reacted at 0 °C for 2 hours. The reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (10 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 4:6 v / v) to give 60 mg (43% yield) of the target compound 5-methyl-1-(2-oxo-2-(pyridin-3-yl)ethyl)-2-(p-tolyl)-1H-pyrrole-3-onitrile as a white solid.

[0641] Step 4. Preparation of 3,8-dimethyl-6-(pyridin-3-yl)pyrrolo[2,1-a]isoquinoline-1-carbonitrile (Example 41)

[0642] The compound prepared in step 3 (50 mg, 0.15 mmol) and trifluoromethanesulfonic acid (112 mg, 0.75 mmol) were dissolved in 1,2-dichloroethane (5 mL) and reacted at room temperature (25 °C) for 2 hours. After the reaction was complete, the reaction solution was diluted with water (10 mL) and extracted with dichloromethane (10 mL × 3). The organic layer was washed with a saturated sodium chloride solution, dried over sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by preparative high-performance liquid chromatography (prep-HPLC) (Gemini 5 µm C18 column, 150 × 21.2 mm, mobile phase: 30%–90% acetonitrile / water containing 0.1% formic acid) to give the final compound 3,8-dimethyl-6-(pyridin-3-yl)pyrrolo[2,1-a]isoquinoline-1-onitrile (10.2 mg, 23% yield), which was a white solid.

[0643] LCMS (ESI-MS): Measured molecular weight C 20 H 15 N3, calculated value 297.1, measured value [M+H]+ = 297.6.

[0644] 1 H NMR (400 MHz, CDCl3): δ 8.95–8.43 (m, 3H), 7.98–7.87 (m, 1H), 7.68–7.53 (m, 2H), 7.50–7.48 (m, 1H), 7.24 (s, 1H), 6.76 (s, 1H), 2.49 (s, 3H), 2.44 (s, 3H).

[0645] <Experimental Example 1> Optical properties of IK15 used to detect Aβ oligomers

[0646] To evaluate the potential of the 41 compounds of the present invention (IK15 compounds: Examples 1 to 13, 22, 26 and 27; and AMD compounds: Examples 14 to 21, 23 to 25 and 28 to 41) as imaging agents for detecting Aβ oligomers, the changes in the fluorescence spectra of IK15 in Aβ monomers (mAβ), Aβ oligomers (oAβ) and Aβ filaments (fAβ) were analyzed.

[0647] To prepare monomeric Aβ samples, Aβ aggregates were treated with 20 mM EPPS (4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid), which dissociates the Aβ aggregates into monomeric forms. To prepare oligomers and filamentous Aβ, Aβ peptides were cultured according to a previously known Aβ aggregation protocol (Lee, HY et al., Amyloid Against Amyloid: Dimeric Amyloid Fragment Ameliorates Cognitive Impairments by Direct Clearance of Oligomers and Plaques, AngewChem Int Ed Engl 62, e202210209 (2023)).

[0648] Emission scanning analysis confirmed that the fluorescence intensity of the candidate compounds mixed with Aβ in IK15o or IK15p was lower than that of the compounds themselves. It was observed that, among the IK15 compounds, excluding the aforementioned substances, the emission spectra of the remaining 14 IK15 compounds were further enhanced upon binding with Aβ monomers, Aβ oligomers, or Aβ filaments. Among these, the emission spectra of IK15a, IK15b, IK15c, IK15d, IK15e, IK15g, IK15j, IK15k, IK15l, and IK15n showed the most significant specific enhancement upon binding with Aβ oligomers. Figure 1A , Figure 1B and Figure 3A ).

[0649] Furthermore, it was confirmed that, similar to IK-15j, AMD-D-255 exhibits a specific enhancement in fluorescence intensity upon binding to Aβ oligomers. It was also confirmed that AMD-D-253, AMD-D-254, AMD-D-257, AMD-D-264, AMD-D-272, AMD-D-273, AMD-D-277, AMD-D-278, AMD-D-279, and AMD-D-280 compounds exhibit low fluorescence intensity when alone, but show a specific enhancement in fluorescence signal upon binding to Aβ monomers, Aβ oligomers, and Aβ filaments. Additionally, it was confirmed that AMD-D-270 is an Aβ-selective fluorescent material, specifically exhibiting fluorescence only when bound to Aβ monomers, Aβ oligomers, and Aβ filaments, while almost no fluorescence is observed when used alone. Figures 1C to 1F ).

[0650] Furthermore, to analyze the fluorescence changes of 16 IK15 substances over time, the fluorescence intensity of candidate compounds mixed with Aβ monomers (mAβ), Aβ oligomers (oAβ), and Aβ filaments (fAβ) was analyzed over 24 hours. This confirmed that the fluorescence intensity decreased over time due to quenching of the fluorescent substances, but when bound to Aβ, the form was stably maintained with a slight decrease in fluorescence intensity. Consistent with previous emission spectral data, a slight decrease in fluorescence intensity was observed when IK15a, IK15b, IK15c, IK15d, IK15e, IK15g, IK15j, IK15k, IK15l, or IK15n substances were bound to Aβ oligomers, while maintaining a stable fluorescence intensity. Figure 2A , Figure 2B and Figure 3B ).

[0651] Furthermore, considering that EPPS might interfere with the interaction between IK15j and Aβ monomers, following the Aβ immobilization plate protocol of Reference Example 2, Aβ monomers and oligomers were immobilized on a culture plate and then incubated with IK15j. Simply immobilizing 10 μM Aβ monomer (mAβ) samples and 10 μM Aβ oligomer (oAβ) samples separately on culture plates, treating them with IK15j (100 μM) for 24 hours, and then measuring the fluorescence intensity, while blank wells were treated only with IK15j (100 μM), without immobilizing the Aβ species. As a result, compared to Aβ monomers, the fluorescence signal of IK15j was significantly enhanced when using Aβ oligomers. Figure 3C These results confirm the existence of a specific interaction between IK15j and the Aβ oligomer.

[0652] <Experimental Example 2> Aβ was detected 1-42 Specificity of IK15 oligomers

[0653] Aβ peptides exhibit variations in length or N-terminal / C-terminal modifications, among which Aβ 1-40 Aβ 1-42 and pyroglutamic acid Aβ 3-42 (Aβ) pE3-42 Aβ is most commonly found in the brains of AD patients. 1-40 and Aβ 1-42 The difference lies in Aβ 1-42 It has two extra residues at its C-terminus. Aβ pE3-42 Through Aβ 1-42 It is produced by multistep protein modification, which converts glutamic acid at the third position of the N-terminus into pyroglutamic acid.

[0654] Therefore, Aβ mutant oligomers were prepared and incubated with IK15j to investigate the selectivity of the compound for the Aβ mutant. Simply adding 10 μM of the oligomer Aβ... 1-40 Sample, 10 μM Aβ 1-42 Samples and 10 μM Aβ pE3-42 Samples were treated with IK15j (10 μM), and the fluorescence intensity of each Aβ mutant sample was measured. The results were compared with those of Aβ... 1-40 and Aβ pE3-42 Compared to oligomers, Aβ 1-42 The fluorescence intensity of IK15j in the oligomer was significantly increased ( Figure 3D ).

[0655] Given that tau protein aggregation is one of the main pathological features of AD, this further confirms the role of IK15j in Aβ. 1-42 The selectivity of the tau domain. Tau protein has four repeat domains, with repeat domain 3 associated with the microtubule-binding domain of tau protein (Le, L. et al., Self-Aggregating Tau Fragments Recapitulate Pathologic Phenotypes and Neurotoxicity of Alzheimer's Disease in Mice, Adv Sci (Weinh), e2302035(2023)). Since repeat domain 3 is the core region of tau aggregates, IK15j is associated with the tau domain or Aβ. 1-42They were incubated together, and changes in fluorescence intensity were compared. Aβ was simply treated separately with IK15j. 1-42 Samples of tau aggregates were analyzed, and fluorescence intensity was measured. Therefore, compared to the tau domain, the signal of IK15j in Aβ... 1-42 Enhancement in the presence of oligomers ( Figure 3E The results described above indicate that IK15j selectively interacts with Aβ. 1-42 Oligomer binding.

[0656] <Experimental Example 3> Identification of Aβ 1-42 IK15 binding site in oligomers

[0657] Aβ aggregates are considered neuropathological biomarkers for Alzheimer's disease (AD), but the transient and polymorphic nature of Aβ makes it difficult to determine the target sites for Aβ imaging probes. To identify Aβ... 1-42 The IK15 binding site of the oligomer was tested according to Reference Examples 2 and 3. Simply, 37 Aβ molecules were prepared. 1-42 Aβ n-(n+5) (n = 1-37) hexameric fragments were obtained and immobilized on culture plates to prepare MAP. This was to investigate whether IK15 targets Aβ. 1-42 Specific sequences or Aβ 1-42 The structure of the oligomer was determined, and two MAP conditions were prepared. First, to confirm the targeting sequence of IK15j, Aβ was prepared in MAP. n-(n+5) + IK15j was treated for 24 hours. Secondly, to induce Aβ-Aβ formation, additional Aβ was... 1-42 The peptide was incubated in MAP for 8 hours. Subsequently, it was treated with IK15j for 24 hours to determine the compound's activity in Aβ. 1-42 Target sites in oligomers ( Figure 3F ).

[0658] The result is consistent with Aβ 1-42 In comparison, in 10 Aβ fragments (Aβ 14-19 To Aβ 23-28 An enhancement of the IK15j signal was observed in ( ) Figure 3G Furthermore, a two-color gradient heatmap was used to show the relationship between IK15j and Aβ. 19-25 Interactions exist between regions (SEQ ID NO.1; FFAEDVG), but the signal of IK15j is relatively low in all segments. Figure 3H This indicates that IK15j is not related to Aβ. 1-42The results described above indicate that the compound does not bind to a specific sequence of Aβ. 1-42 The specific sequences form interactions, therefore IK15j cannot recognize monomeric Aβ. 1-42 .

[0659] In addition, the effect of IK15j on Aβ was measured. 1-42 Fragments and with Aβ 1-42 The fluorescence intensity changes of each complex were observed. As a result, changes in fluorescence intensity were observed in Aβ. 1-42 -Aβ n-(n+5) Under the +IK15j condition, the overall signal of IK15j increases, indicating a structural interaction between IK15j and the Aβ oligomer. This compound interacts with Aβ... 1-42 and Aβ as two main structural domains 19-25 (SEQ ID NO.1; FFAEDVG) and Aβ 35-42 The complex of (SEQ ID NO.2; MVGGVVIA) is firmly bound ( Figure 3I and Figure 3J ).

[0660] The central hydrophobic region containing the KLVFFA domain is known to be Aβ. 1-42 The most readily aggregated sequence is the GGVVIA domain, which is a hydrophobic C-terminal core that stabilizes the hydrophobic interactions of Aβ. This confirms that IK15j targets the hydrophobic core region of the Aβ oligomer. In particular, the compound exhibits appropriate binding strength to the N-terminus of Aβ. The hydrophilic region of Aβ is known to be... 1-8 (SEQ ID NO.3; DAEFRHDS) alters the aggregation tendency of Aβ and affects the secondary structure of Aβ oligomers. IK15j can interact with both the hydrophobic and amphiphilic regions of the Aβ oligomer, forming a stable interaction with it. In summary, these results indicate that IK15j interacts structurally with the Aβ oligomer, and that the compound specifically targets the hydrophobic core of Aβ aggregation and the hydrophilic N-terminus of the Aβ oligomer.

[0661] <Experimental Example 4> Soluble Aβ oligomers were detected in brain tissue lysates, cerebrospinal fluid, and plasma of a 5XFAD mouse model.

[0662] 5XFAD is a transgenic AD mouse model that expresses human APP and presenilin-1 and exhibits the main pathological features of AD, including the formation of Aβ aggregates in the brain.

[0663] The experimental protocol for IK15 ex vivo using the 5XFAD mouse model is as follows: Figure 4AAs shown. Specifically, as described in Reference Example 4, soluble Aβ species were detectable as early as 2 months after birth in the 5XFAD transgenic mouse model, and their levels increased significantly with age. Therefore, 6-month-old female 5XFAD transgenic mouse models (n=3) and age-matched female wild-type mice (n=3) were prepared. Subsequently, each mouse was sacrificed, and brain tissue lysates (cortex and hippocampus), cerebrospinal fluid, and plasma were obtained according to the methods described in Reference Examples 5 to 7.

[0664] Brain tissue lysates (cortex and hippocampus) and cerebrospinal fluid from 5XFAD mouse models or wild-type mice were treated with each of the 16 IK15 example compounds at 50 μM. Each sample was then placed into the wells of a 96-well half-region black microplate. The total volume of each sample was 100 μL. After treatment with the example compounds for 24 hours, the unique excitation / emission values ​​of each of the four substances were scanned to confirm changes in fluorescence signal, and the fluorescence scan results were detected using a microplate reader.

[0665] The results showed that in cortical analyses treated with IK15d, IK15g, IK15j, IK15q, IK15r, or IK15s, significant differences in fluorescence intensity were observed between the 5XFAD transgenic mouse cortical group and the normal mouse cortical group. In analyses using IK15d, IK15j, IK15q, IK15r, and IK15s, the fluorescence intensity of the 5XFAD transgenic mouse cortex was enhanced; in analyses using IK15g, the fluorescence intensity of the normal mouse cortex was enhanced. This method confirmed that differences could be distinguished between 5XFAD transgenic mice and normal mice in cortical analyses using a total of six IK15 substances (IK15d, IK15g, IK15j, IK15q, IK15r, or IK15s). Figure 4B , Figure 5A and Figure 5B ).

[0666] Furthermore, in hippocampal analyses treated with IK15a, IK15b, IK15j, IK15k, IK15n, IK15r, or IK15s, significant differences in fluorescence intensity were observed between the 5XFAD transgenic mouse hippocampus and the normal mouse hippocampus. In analyses using IK15a, IK15b, IK15j, IK15k, IK15r, or IK15s, fluorescence intensity was enhanced in the hippocampus of 5XFAD transgenic mice; in analyses using IK15n, fluorescence intensity was enhanced in the hippocampus of normal mice. This method confirms that differences can be distinguished between 5XFAD transgenic mice and normal mice in hippocampal analyses using a total of seven IK15 substances (IK15a, IK15b, IK15j, IK15k, IK15n, IK15r, or IK15s). Figure 4C , Figure 6A and Figure 6B ).

[0667] Furthermore, significant differences in fluorescence intensity were observed between 5XFAD transgenic mice and normal mice in cerebrospinal fluid analysis treated with IK15a, IK15j, IK15n, or IK15r. In analyses using IK15a, IK15j, or IK15r, fluorescence intensity was enhanced in the cerebrospinal fluid of 5XFAD transgenic mice; in analyses using IK15n, fluorescence intensity was enhanced in the cerebrospinal fluid of normal mice. This method confirms that differences can be distinguished between 5XFAD transgenic mice and normal mice in cerebrospinal fluid analysis using a total of four IK15 substances (IK15a, IK15j, IK15n, or IK15r). Figure 4D , Figure 7A and Figure 7B ).

[0668] To analyze the relationship between Aβ oligomers and IK15 substances in plasma using a total of four IK15 substances (IK15a, IK15j, IK15r, and IK15s), plasma samples were prepared from 6-month-old female 5XFAD transgenic (TG) mouse models (n=3) and age-matched female normal (WT) mice (n=3). The four IK15 substances (IK15a, IK15j, IK15r, and IK15s) were selected in the previous experiments as having high binding capacity to Aβ oligomers. After collecting each plasma sample, it was treated with EDTA and then with each IK15 substance (50 μM). Each sample was then added to the wells of a 96-well half-zone black microplate. The total volume of each sample was 50 μL. To determine the changes in fluorescence signal after 24 hours, the unique excitation / emission values ​​of each of the 16 substances were scanned, and the fluorescence scan results were detected using a microplate reader.

[0669] The results showed significant differences in fluorescence intensity between the 5XFAD transgenic mouse group and the normal mouse group in plasma analysis using IK15a, IK15j, or IK15r of the four candidate substances. With IK15a, fluorescence intensity was enhanced in the plasma of normal mice; conversely, with IK15j and IK15r, fluorescence intensity was enhanced in the plasma of 5XFAD transgenic mice. This method confirms that differences can be distinguished between 5XFAD transgenic mice and normal mice in plasma analysis using a total of three IK15 substances (IK15a, IK15j, or IK15r). Figure 4E and Figure 8 ).

[0670] In addition, to confirm whether IK15j precisely targets Aβ oligomers, brain staining was performed in the hippocampus and cortical regions using IK15j and anti-Aβ antibody 6E10, as described in Reference Example 8, to visualize the colocalization of the signal. Figure 4F ).

[0671] The result, such as Figure 4F and 4G As shown, no IK15j signal was observed in the hippocampus or cortical region, confirming that IK15j cannot detect insoluble plaques in the brains of 5XFAD mice. Figure 4G ).

[0672] This method confirmed that IK15j can selectively detect Aβ oligomers in brain tissue lysates, cerebrospinal fluid, and plasma from a 5XFAD mouse model.

[0673] <Experimental Example 5> confirmed the interaction between IK15 and plasma Aβ.

[0674] To investigate whether the compounds of the present invention interact with Aβ oligomers in plasma, as described in Example 9, Aβ oligomers were spiked into human plasma (100, 10, 1, 0.1, 0.01, 0.001, 0 pg / mL) and treated with IK15j (100 μM) for 8 hours.

[0675] The results showed that, compared with blank human plasma (0 pg / mL), the IK15j signal was significantly enhanced in spiked samples at 1 pg / mL. Compared with the control group, the fluorescence intensity of Aβ-spiked samples at 10-100 pg / mL was increased by 52% ( Figure 9A ).

[0676] <Experimental Example 6> confirms the interaction between the compound of the present invention and plasma proteins.

[0677] The solid portion of plasma, excluding water, consists of coagulants and plasma proteins. Albumin is the most abundant protein in plasma, and fibrinogen is the main coagulant. Considering that fibrinogen or albumin may interact with AD diagnostic tools and interfere with the selectivity of compounds, in order to analyze the selectivity of the compounds of the present invention for such proteins, as described in Reference Example 10, IK15j (25 μM) and Aβ were... 1-42 The sample was incubated with albumin or fibrinogen, and the fluorescence signal of each sample was analyzed.

[0678] Therefore, compared to other proteins, Aβ 1-42 The strength of IK15j was significantly enhanced in the oligomers. Conversely, studies have found that IK15j does not interact with fibrinogen or albumin. Figure 9B ).

[0679] In addition, to investigate whether albumin interferes with IK15j and Aβ 1-42 The binding between oligomers, as described in Reference Example 11, involves IK15j and Aβ. 1-42 The oligomers were incubated with albumin, and the fluorescence signal of each sample was analyzed.

[0680] The results showed that, compared with Aβ alone... 1-42 Compared to oligomer samples, Aβ 1-42 The signal intensity decreased in both oligomer and albumin samples, but the difference was not significant. Figure 9C ).

[0681] Furthermore, to confirm that albumin does not mask the target site of IK15j, Aβ oligomer-albumin complexes and Aβ oligomers immobilized on a culture plate were prepared as described in Example 11. Subsequently, IK15j (25 μM) was added to both wells for treatment, and signal differences were compared. Figure 9D ).

[0682] Therefore, no significant difference in fluorescence intensity was observed between the Aβ oligomer-albumin complex and the Aβ oligomer. This confirms that IK15j interacts with both the Aβ oligomer and the Aβ oligomer-albumin complex in plasma, indicating that albumin does not mask the target site of IK15j. Figure 9E ).

[0683] The above description of the present invention is for illustrative purposes only. Those skilled in the art should understand that the present invention can be readily modified into other specific forms without altering its technical concept or essential features. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

Claims

1. A compound represented by the following chemical formula 1, or a salt thereof: [Chemical Formula 1] In the chemical formula 1, The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

2. The compound or a salt thereof according to claim 1, wherein, The R 1a and R 1b Each can be independently hydrogen, deuterium, or halogen; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-5 Alkyl, substituted or unsubstituted C 2-5 alkenyl, substituted or unsubstituted C 2-5 Alkyne, substituted or unsubstituted C 1-5 Alkoxy, substituted or unsubstituted 6-membered aryl, or substituted or unsubstituted 6-membered heteroaryl, or said R 1c and R 1d They connect to each other to form the compound represented by chemical formula 2. The X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It can be hydrogen, deuterium, or halogen; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, substituted or unsubstituted C. 1-5 Alkyl, substituted or unsubstituted C 2-5 alkenyl, substituted or unsubstituted C 2-5 Alynyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-5 Alkyl), substituted or unsubstituted C 1-5 Alkyl or substituted or unsubstituted 6- to 10-membered aryl groups.

3. The compound or a salt thereof according to claim 1, wherein, The R 1a and R 1b Each can be either hydrogen or halogen; The R 1c and R 1d Each is independently hydrogen, halogen, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted C 1-5 Alkoxy or substituted or unsubstituted 6-membered aryl groups, or The R 1c and R 1d They connect to each other to form the compound represented by chemical formula 2. The X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), The R 4 It is hydrogen; The R 2a and R 2b Each is independently hydrogen, halogen, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 6-membered heteroaryl; The R 3a and R 3b Each can be independently represented as hydrogen, -CN, -C(O)NH2, or -C(O)O(C 1-5 Alkyl), substituted or unsubstituted C 1-5 Alkyl or substituted or unsubstituted 6-membered aryl groups.

4. A method for preparing a compound represented by the following chemical formula 1, as shown in the following reaction formula 1, comprising the following steps: Compound 1001 and compound 1002 were reacted in an organic solvent to obtain compound 1003 (step 1). Compound 1003 and NH4OAc were reacted in an organic solvent to obtain compound 1004 (step 2). Compound 1005-1, Compound 1005-2 or Compound 1005-3, and Compound 1004 are reacted in an organic solvent to obtain Compound 1006 (Step 3); and Compound 1006 is reacted with a catalyst in an organic solvent to obtain the compound represented by the following chemical formula 1 (Compound 1007) (Step 4). [Reaction Formula 1] In the reaction formula 1, The R is R 1c or X q ; The X p and X q Each can be independently hydrogen, deuterium, or halogen; The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

5. The preparation method according to claim 4, The organic solvents used in steps 1 to 4 are each independently selected from one or more of the group consisting of methanol, ethanol, propanol, acetonitrile, dichloroethane, ethyl acetate, dimethyl sulfoxide, and dimethylformamide.

6. The preparation method according to claim 4, The catalyst in step 4 is selected from one or more of the group consisting of trifluoromethanesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, and scandium trifluoromethanesulfonate.

7. A composition for detecting β-amyloid protein, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the chemical formula 1, The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

8. The composition for detecting β-amyloid protein according to claim 7, wherein, The Aβ is selected from one or more of the group consisting of Aβ oligomers, Aβ fibrils, Aβ fibers, and Aβ plaques.

9. The composition for detecting β-amyloid protein according to claim 7, wherein, The Aβ contains mutants of Aβ.

10. The composition for detecting β-amyloid protein according to claim 7, wherein, The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof interacts with one or more domains selected from the group consisting of: a domain containing the amino acid sequence of SEQ ID NO. 1, a domain containing the amino acid sequence of SEQ ID NO. 2, and a domain containing the amino acid sequence of SEQ ID NO.

3.

11. A method for detecting β-amyloid protein, comprising the following steps: Contact the isolated biological sample with a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the chemical formula 1, The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

12. The method for detecting β-amyloid protein according to claim 11, wherein, The isolated biological samples are selected from one or more of the group consisting of blood, plasma, cerebrospinal fluid, cortical lysates, and hippocampal lysates.

13. A composition for diagnosing neurodegenerative diseases, comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the chemical formula 1, The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

14. A method for providing information on the diagnosis of neurodegenerative diseases, comprising the following steps: Contact the isolated biological sample with a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the chemical formula 1, The R 1a and R 1b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 1c and R 1d Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl, or substituted or unsubstituted 5- to 10-membered heteroaryl, or the R 1c and R 1d They connect to form the compound represented by the following chemical formula 2. [Chemical Formula 2] In the chemical formula 2, X 1 X 2 X 3 and X 4 Each independently is C(R) 4 ), nitrogen, oxygen, sulfur or phosphorus, The R 4 It is hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 2a and R 2b Each is independently hydrogen, deuterium, halogen, amino, hydroxyl, substituted or unsubstituted C. 1-10 Alkyl, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted C 2-10 Alkyne, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl; The R 3a and R 3b Each can be independently hydrogen, deuterium, halogen, amino, hydroxyl, -CN, -C(O)NH2, -C(O)O(C 1-10 Alkyl), substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted 6- to 10-membered aryl or substituted or unsubstituted 5- to 10-membered heteroaryl.

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