WO2009054653A2 - Isoindolones with high binding affinity to beta-amyloid aggregates and fibrils, and its use and preparation method - Google Patents

Abstract

The present invention relates to isoindolone compounds having high binding affinity to beta-amyloid fibrils, thereby being useful for an early diagnosis of degenerative cerebral diseases including dementia, and prevention and treatment of such diseases, its use and a method for preparing the same.

Description

ISOINDOLONES WITH HIGH BINDING AFFINITY

TO BETA-AMYLOID AGGREGATES AND FIBRILS. AND

ITS USE AND PREPARATION METHOD

TECHNICAL FIELD

The present invention relates to isoindolones, which have high binding affinity to beta-amyloid fibrils and thus are useful for an early diagnosis of degenerative cerebral diseases including dementia, and for the prevention and treatment thereof, its preparation method and a pharmaceutical composition comprising the same.

BACKGROUND ART

According to the continuous improvement of living standards and the development of health and medical techniques, the average life span of people lengthens and the population of elderly people is rapidly increasing. As the population structure is aging, a resultant natural health problem, in particular, senile dementia, a degenerative neurological disorder frequently observed with the aged people, emerges as a major social problem as one of the most serious geriatric diseases.

Dementia is a clinic syndrome appearing due to various causal diseases, not a disease resulting from a single cause, and up to now, 70 or more causal diseases have been known. Among them, the causal disease known to induce the senile dementia is Alzheimer's disease, cerebrovascular dementia, Parkinsons disease, etc.

Alzheimer's disease (AD) takes the largest portion of the causal diseases of dementia, which was first discovered by Alois Alzheimer, a Germany doctor, in 1907. Alzheimer's disease, a degenerative neurological disorder, has the risk increasing as people get older. The statistics in the United States shows that one of ten people over the age of 65 and four of ten people over the age of 85 suffer Alzheimer's disease.

Patients suffering from Alzheimer's disease are subject to reduction in the volume of cerebral tissues and cerebral activity metabolism, and the size of brain of such patients is merely about one-fourth because the number of nerve cells of the cerebrum decreases. Its symptom includes a loss of memory, a loss of perception, a disorder of thinking and judgment, a change in humanity, a feeling disorder, etc. At a final stage, the patients cannot take care of themselves to end in death. The accurate cause of Alzheimer's disease or its treatment method has not yet been found, but it can be found that the brains of the patients show that the nerve cells secreting acetylcholine, which is a neurotransmitter, are selectively degenerated.

Acetylcholine is a substance critically used for a brain function such as memory and learning, and in the past, it has been believed that the reduction of acetylcholine induces Alzheimer's disease. Thus, drugs having various mechanisms for resolving the deficiency problem of acetylcholine have been developed. Representative drugs thereof are Tacrine (product name: Cognex), Donepezil (product name: Aricept), Rivastigmine (product name: Exelong), etc., which were approved by FDA of the United States, estrogen that can inhibit or prevent nerval denaturalization, antioxidant such as vitamin C or E, anti-inflammatory drugs such as non-steroidal anti-inflammatory drug (NSAID), etc. However, theses drugs are inhibitors of acetylcholine breakdown enzyme, which cannot be a fundamental treatment means and have a limitation in that they merely improve a damaged perceptive function temporarily. Thus, it is necessary to develop drugs allowing causal treatment.

Pathological opinions appearing in relation to Alzheimer's disease as known up to now may include senile plaques (SPs) (or neuritic plaques), a protein deposit observed outside the nerve cells, and neurofibrillary tangles (NFTs) seen as an entangled thread bundle within the nerve cells. Most of the senile plaques are formed with beta-amyloid protein consisting of about 40-42 amino acids, and the neurofibrillary tangles are formed by intra-cell protein aggregation due to hyperphosphorylation of tau, which is microtubule-binding protein. It is assumed that abnormal protein aggregation commonly identified in the above two phenomena has a close relationship with the development of diseases. Thus, a beta-amyloid hypothesis that beta-amyloid would act as a cause of developing Alzheimer's disease has been strongly admitted according to various experimental evidences, after the acetylcholine hypothesis that reduction of acetylcholine was a cause of Alzheimer's disease in early 1980s. The senile plaques have one or two types of proteins entangled at its central portion. The proteins consist of 40 or 42 amino acids. The protein consisting of 40 amino acids is called as Aβ40, and the protein consisting of 42 amino acids is called as Aβ42. Aβ40 has little virulence, while Aβ42 has much virulence and it is a major cause of Alzheimer's disease because it takes most of the amyloid deposition existing at the central portion of the plaques.

As for a generation process of Aβ42, first, beta-amyloid protein is generated from amyloid precursor protein (APP). α-, β-, and γ-secretases exist in brains, and Aβ42 is generated when the β-, and γ-secretases decompose both ends of Aβ42 amino acid sequence of APP. APP is a protein present in the brains of normal people, and it is mostly metabolized by α-secretase and mostly exists as secretor-type sAPPα. sAPPα acts like a growth factor in brains, accelerates the growth of brain cells, and plays a key role for memory and learning ability. In case of the patients of Alzheimer's disease, such metabolism becomes abnormal, by which Aβ42, which should exist in a small amount, is generated in a large amount to form fibrils, and the fibrils gather to form plaques.

The significance of medical images in today's clinical medicines is hugely increasing, and the medical images checking such as X-ray, sonogram, computer tomography (CT), etc. have remarkably contributed to the development of the 20 century medical science, and diagnoses and treatments of patients. However, since such anatomic image technology has been aimed at an early discovery of an anatomic change, it has a limitation on discovering an abnormality at the level of molecules or cells and on applying it to treatment. Recently, a gene therapy and a cloning technique have been developed depending on the development of molecular biology, and molecular images allowing to image the change of a molecule or cell level in vivo have received much attention since a reporter gene technology was introduced in 1990.

Molecular imaging is to image life phenomena of living organisms in units of cells or molecules with a non-invasive method. The difference of a minute function can be imaged at an initial stage with no anatomic change according to a disease, which enables to help diagnose a disease. Thus, molecular imaging allows early discovery of a pre-disease stage and treatment the same, proposes a new possibility of developing treatment drugs, and allows early evaluation of reaction on the treatment, by which customized treatment suitable for each patient can be provided with minimized toxicity of the treatment. Inspecting methods for obtaining such images include a single photon emission computed tomography (SPECT) and positron emission tomography (PET) using a radioactive element. In order to evaluate the function of the central nerve system, the imaging technique using the PET or SPECT has been developed at a quite fast pace, and it is actually useful technique for basic medical researches and a clinical medicine. These two techniques are all useful for discriminating a normal condition and information on diseases. In general, the PET is more useful for quantitative analysis because it has a better sensitivity and resolution, and reflects better a biochemical change. Recently, researches on the brain images of dementia patents are actively ongoing. Among them, anatomic images such as CT, MRI, etc. are capable of discovering structural abnormalities of brains but have a limitation in that it cannot sensitively discover a typical degenerative change in the brains at an early stage. Thus, molecular images using a nuclear medical technique such as SPECT or PET have been actively studied as a tool for diagnosing Alzheimer's disease. In particular, researches have been actively ongoing to develop a radioactive probe for PET to image beta-amyloid accumulation which has been believed as a cause material of Alzheimer's disease. Compounds known to be well bound with β-amyloid fibrils include

Congo red (CR) derivative, benzothiazole derivative, naphthalene derivative, etc. The CR derivative is a dye strongly binding to β-amyloid fibrils, but it cannot pass through brain blood barrier (BBB) because it has organic carboxyl groups which are too much polar and hydrophilic. Thioflavin-T (Th-T), a benzothiazole derivative, also has a difficulty in passing through the brain blood barrier because it is charged with positive and negative ions, and thus, there is a problem to be developed as a radioactive probe.

In order to resolve this problem, recently, studies on the derivatives of the compounds have been actively conducted. Among them, representative compounds such as N-methyl-[¹¹C]2-(4'-methylaminophenyl) 6-hydroxy- benzothiazole [Pittsburgh Compound-B (PIB compound)] [See Klunk, W. E. et al., Annals of Neurology, 55, 306 (2004)], 2-(4'-dimethylamino- phenyl)-6-iodoimidazo[1 ,2-a]pyridine (IMPY compound) [See Kung, H. F. et al., Brain Research, 956, 202 (2002)], 2-(4'-dimethylaminophenyl)- 6-iodobenzoxazole (IBOX compound) [See Kung, H. F. et al., Nuclear Medicine and Biology, 28, 887 (2001 )], (E,E)-1-iodo-2,5-bis(3-hydroxy- carbonyl-4-methoxy)styrylbenzene (IMSB compound) [See Kung, H. F. et al., Journal of Medicinal Chemistry, 44, 2270 (2001 )]), etc., were reported to be well bound with β-amyloid and easily pass through the brain blood barrier. Because these compounds specifically recognize β-amyloid fibrils, they have been much studied as a reagent for an early diagnosis to diagnose dementia before the formation of the senile plaques or the β-amyloid fibrils in brains.

PIB, a benzothiazole derivative, which is obtained by chemically changing Th-T so as to increase liposolubility, can pass through the brain blood barrier and strongly bound with β-amyloid, so that it can be used as a diagnosing reagent. However, because PIB is labeled with the radioactive isotope ¹¹C, its half-life is very short as 20 minutes. Thus, its practicability is low, which results in limitation on its use. 2-(1-(6-[(2-[¹⁸F]- fluoroethyl)- (methyl)amino]-2-naphthyl}ethylidene}malononitrile (FDDNP), which is a naphthalene derivative, is the first hydrophobic molecular image probe developed by chemically changing naproxen. This compound has been reported that it can pass through the brain blood barrier, has a high affinity to β-amyloid fibrils and makes it possible to detect the amount and position of the senile plaques (SPs) and neurofibrillary tangles (NFTs). However, FDDNP has shortcomings that it is not specifically bound only to the β-amyloid but nonspecifically bound to other tissues.

The cause of the Alzheimer's disease has not yet been clearly discovered up to date, but when various evidences are generalized, accumulation of β-amyloid, a virulent protein and the resultant degeneration of nerve cells have been known to be a typical symptom or cause. Recently, it has been reported that oligomers are virulent more than five times compared with the β-amyloid fibrils and it has been discovered that only Aβ42 forms such oligomers. Thus, oligomers, as well as fibrils generated by Aβ42, emerge as new targets for treating Alzheimer's disease.

DISCLOSURE OF THE INVENTION

Therefore, it is an object of the present invention to provide a compound with high binding affinity to beta-amyloid fibrils or its pharmaceutically acceptable salt, and a preparation method.

In addition, it is another object of the present invention to provide a pharmaceutical composition for diagnosing dementia at an early stage, or for preventing or treating the dementia, comprising said compound as an effective component.

In addition, it is still another object of the present invention to provide an agent for inhibiting formation of a beta-amyloid fibrils, comprising said compound as an effective component.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the detection results of the beta-amyloid fibril formation using fluorescence of thioflavin-T (Th-T). FIG. 2 is a graph showing TZDM dissociation constant (Kd) values which were obtained using beta-amyloid fibrils and 2-(4'-dimethylamino- phenyl)-6-[¹²⁵ l]iodobenzothiazole (¹²⁵I-TZDM).

MODES FOR CARRYING OUT THE PREFERRED EMBODIMENTS The inventors of the present invention discovered that isoindolone compounds including isoindol-1 ,3-diones and isoindol-1-ones, which are new compounds synthesized based on an structure of indoprofen, non-steroidal anti-inflammatory drugs, exhibit high binding affinity specifically to beta-amyloid and easy penetration brain blood barrier. In addition, the inventors of the present invention completed the invention by discovering that said compounds can inhibit formation of fibrils or oligomers, as well as plaques, so that they can be useful for the prevention or treatment of the diseases related to accumulation of beta-amyloid and can be used for an early diagnosis of such diseases if they are labeled with a radioactive isotope, for example, ¹¹C and ¹⁸F.

Therefore, the present invention relates to a compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof:

Formula 1 :

wherein A is C=O or CH₂ group; Ri is H, OH, a halogen atom selected from fluorine, chlorine, bromine and iodine, a CrC₈ alkyl group, a Ci-Cs alkoxy group, a tosyloxy-CrC₈ alkoxy group, a mesyloxy-d-Cs alkoxy group, a nosyloxy-d-Cβ alkoxy group, CrC₈ alkoxy group substituted with halogen atoms selected from fluorine, chlorine, bromine and iodine, or a CrCe alkylamino group; R₂ is H, a halogen atom selected from fluorine, chlorine, bromine and iodine, or a CrC₈ alkyl group; and R₃ and R₄ are independently H, a CrC₈ alkoxy group or a CrCs alkylamino group.

The compound of Formula 1 includes a compound labeled with a radioactive element, for example, a compound in which one or more halogen or carbon atoms within its molecular structure are radioactive isotopes, for example, ¹⁸F Or ¹¹C.

In addition, the present invention provides a preparation method of the compound of Formula 1 and a pharmaceutical composition for treating a degenerative cerebral disease, comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an effective component. In addition, present invention provides a pharmaceutical composition for preventing, treating and early diagnosing a disease directly related to beta-amyloid fibrils, comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an effective component and.

When the pharmaceutical composition is used as a diagnosing agent, the compound of Formula 1 may be a compound labeled with a radioactive element, for example, a compound in which one or more halogen or carbon atoms within its molecular structure are radioactive isotopes, for example, ¹⁸F

Or ¹¹C.

The present invention will be described in more detail below. The following Formula 1a represents isoindol-1 ,3-dione derivatives that

A of the compound of Formula 1 is a C=O group: Formula 1a:

wherein Ri, R₂, R₃, and R₄ are the same as defined in Formula 1.

The following Compounds 1a-1 to 1a-27 are preferred embodiments of the isoindol-1 ,3-dione derivative compound represented by Formula 1a.

(1 a-1 ) 5-dimethylamino-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1 a-2) 5-methoxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1 a-3) 5-hydroxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1a-4) 5-methyl-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

(1 a-5) 2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1a-6) 5-fluoro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1 a-7) 5-chloro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1a-8) 5-bromo-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; (1a-9) 5-dimethylamino-2-(4-dimethylamino-3-methoxyphenyl)isoindol-

1 ,3-dione;

(1a-10) 5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

(1a-11 ) 5-methyl-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

(1a-12) 2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-ione; (1 a-13) 5-fluoro-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

(1a-14) 5-chloro-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

(1a-15) 5-bromo-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

(1 a-16) 2-(3-dimethylamino-4-methoxyphenyl)isoindol-1 ,3-dione; (1 a-17) 2-(3<limethylaminophenyl)isoindol-1 ,3-dione; (1a-18) 5-methoxy-2-(4-aminophenyl)isoindol-1 ,3-dione; (1 a-19) 5-methoxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione; (1a-20) 5-hydroxy-2-(4-aminophenyl)isoindol-1 ,3-dione;

(1 a-21 ) 5-hydroxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione; (1 a-22) 5-(2-fluoroethoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione; (1 a-23) 5-(2-fluoroethoxy)-2-(4-methylaminophenyl)isoindol-1 ,3-dione; (1 a-24) 5-(2-fluoroethoxy)-2-(4-dimethylaminophenyl)isoindol-1 ,3- dione;

(1 a-25) 5-(3-fluoropropoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione; (1 a-26) 5-(3-fluoropropoxy)-2-(4-methylaminophenyl)isoindol-1 ,3- dione;

(1a-27) 5-(3-fluoropropoxy)-2-(4-dimethylaminophenyl)isoindol-1 ,3- dione.

The following Formula 1b represents isoindol-1-one derivatives in which A of the compound of Formula 1 is CH₂ group: Formula 1b:

wherein R-i, R₂, R₃, and R₄ are the same as defined in Formula 1.

The following Compounds'! b-1 to 1b-33 are preferred embodiments of the isoindol-1-one derivative compound represented by Formula 1b:

(1 b-1 ) 5-methoxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1- one;

( 1 b-2) 5-hydroxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1 -one; (1b-3) 5-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

(1b-4) 2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1 b-5) 5-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1b-6) 5-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1b-7) 5-bromo-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1 b-8) 6-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

(1b-9) 6-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1 b-10) 6-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; (1 b-11 ) 5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one; (1 b-12) 5-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1 b-13) 2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol- 1-one;

(1 b-14) 5-fluoro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1 b-15) 5-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1 b-16) 5-bromo-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1 b-17) 6-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one; (1 b-18) 6-fluoro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1b-19) 6-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one;

(1 b-20) 2-(3-dimethylamino-4-methoxyphenyl)-2,3-dihydroisoindol- 1-one;

(1b-21 ) 2-(3-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

(1 b-22) 5-methoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one;

(1 b-23) 5-methoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1-one;

(1b-24) 5-methoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one; (1 b-25) 5-methoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1 -one;

(1 b-26) 5-(2-fluoroethoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one;

(1 b-27) 5-(2-fluoroethoxy)-2-(4-methylaminophenyl)-2,3-dihydro- isoindol-1-one;

(1b-28) 5-(2-fluoroethoxy)-2-(4-dimethylaminophenyl)-2,3-dihydro- isoindol-1-one; (1b-29) 5-(3-fluoropropoxy)-2-(4-aminophenyl)-2,3-dihydroisoindol- 1-one;

(1 b-30) 5-(3-fluoropropoxy)-2-(4-methylaminophenyl)-2,3-dihydro- isoindol-1-one; (1 b-31 ) 5-(3-fluoropropoxy)-2-(4~dimethylaminophenyl)-2,3-dihydro- isoindol-1-one;

(1b-32) 5-methoxy-2-(4-¹¹methylaminophenyl)-2,3-dihydroisoindol- 1-one;

(1 b-33) 5-(2-¹⁸fluoroethoxy)-2-(4-dimethylaminophenyl)-2,3-dihydro- isoindol-1-one.

The compound of Formula 1 includes any type of pharmaceutically acceptable salts derived from a conventional inorganic or organic acid well known to the skilled person in the art. The examples of the inorganic or organic acid which may form pharmaceutically acceptable salt with the compound of Formula 1 may include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, etc.

A preparation method of the compound of Formula 1 according to the present invention will be described below.

The preparation method of the compound of Formula 1 according to the present invention comprises a step of reacting a phthalic anhydride, for example, a compound represented by Formula 2a below, or an ester of alkylbenzoic acid having an appropriate leaving group, for example, a compound represented by Formula 2b below, with an aromatic amine compound represented by Formula 3 below: Formula 2a:

Formula 2b:

Formula 3:

wherein R-i, R₂, R₃ and R₄ are respectively the same as defined in

Formula 1.

The following Reaction Scheme 1 illustrates the preparation method of the compound of Formula 1 according to the present invention in which the compound of Formula 2a or 2b is reacted with the compound of Formula 3 to prepare the compound of Formula 1 : Reaction Scheme 1 :

2b wherein A and Ri, R₂, R₃ and R₄ are respectively the same as defined in Formula 1.

The reaction shown in Reaction Scheme 1 is preferably carried out with heating and refluxing in an acidic solvent. As the acidic solvent, acetic acid, hydrochloric acid or p-toluenesulfonic acid, and preferably acetic acid may be used. The compound of Formula 2a or 2b used in the present invention as a starting material may be prepared according to the following Reaction Schemes 2a and 2b with a known method [See Kazemi, F. et al, Phosphorus, Sulfur and Silicon, 2003, 178, 2287-2291 ; Prasad, C. S. N. et al, Heterocycl. Commun. 2002, 8, 281-286]) from a known compound phthalic acid or 2-methylbenzoic acid:

Reaction Scheme 2a:

2]octane

Reaction Scheme 2b:

wherein Ri and R₂ are respectively the same as defined in Formula 1.

As shown in Reaction Scheme 2a, phthalic acid is cyclized starting from 0⁰C to a room temperature in the presence of thionyl chloride, phosphorus oxychloride or phosphorus pentachloride, and 1 ,4-diazabicyclic- [2.2.2]octane in a suitable solvent, for example, dichloromethane or chloroform, so as to obtain the compound of Formula 2a. Alternatively, as shown in Reaction Scheme 2b, 2-methylbenzoic acid is refluxed with thionyl chloride, phosphorus oxychloride or phosphorus pentachloride in an alcohol, preferably, methanol or ethanol, to obtain ethyl 2-methyl benzoate compound, and the ethyl 2-methyl benzoate compound is refluxed in the presence of N-bromosuccinimide and benzoylperoxide in a suitable organic solvent so as to brominate to obtain the ethyl 2-bromoethyl benzoate compound of Formula 2b.

The following Reaction Scheme 3 shows a method for preparing Compounds 1a-18, 1a-20, 1b-22, and 1 b-24, the examples of the compounds of Formula 1 of the present invention from the compound of Formula 2a or 2b in which R2 is hydrogen and Ri is a methoxy group: Reaction Scheme 3:

wherein A and R₄ are respectively the same as defined in Formula 1.

As shown in Reaction Scheme 3, the compound of Formula 2a or 2b is reacted with the compound of Formula 3 to obtain a 5-methoxy-2-(4-nitrophenyl)isoindol-1 ,3-dione or 5-methoxy-2-(4-nitrophenyl)- 2,3-dihydroisoindol-1-one compound, and this compound is reacted with tin (Sn), zinc (Zn), iron (Fe), hydrazine (H₂N₄/FeCI₃), copper sulfate (CuSO₄) or tin chloride (SnCI₂), preferably, with tin chloride (SnCI₂), at 64°C to 78°C for 8 to 12 hours in an alcohol solvent such as methanol, ethanol, etc., under an acidic condition, to obtain Compounds 1a-18 and 1b-22.

In addition, boron trichloride, boron, trifluroide, boron tribromide, boron triiodo or iodotrimethylsilane, preferably, boron tribromide is added to Compounds 1a-18 and 1 b-22, respectively, and then stirred for 8 to 12 hours at room temperature to 70⁰C for demethylation, to obtain Compounds 1a-20 and 1 b-24.

The following Reaction Scheme 4 illustrates a representative method for preparing the compounds of Formula 1 in which R₃ is an alkylamino group, i.e., Compounds 1a-19, 1a-21 , 1b-23, and 1b-25. This reaction process comprises a reductive amination of the amino compound obtained from the reduction step of a nitro group as illustrated in Reaction Scheme 3 with sodium methoxide, paraformaldehyde and sodium borohydride at O⁰C to 65°C for two or three hours:

Reaction Scheme 4:

wherein A and R₄ are respectively the same as defined in Formula 1 , and R₅ is a methoxy or hydroxyl group.

The compounds of Formula 1 in which R₃ or R₄ is respectively a dialkylamino group, i.e., Compounds 1a-1 to 1a-17 and 1 b-1 to 1 b-21 , can be directly prepared according to Reaction Scheme 1.

The following Reaction Scheme 5 illustrates a preparation process of Compounds 1a-22 to 1a-27 and 1 b-26 to 1b-31 of the present invention. This reaction comprises a nucleophilic substitution of the 5-hydroxy- isoindol-1 ,3-dione or 5-hydroxy-2,3-dihydroisoindol-1-one compound obtained by the method as illustrated in Reaction Scheme 3 with a compound of a general formula F(CH₂)n0R₆ (wherein Re is a tosyl (Ts), mesyl (Ms), or nosyl (Ns) group, and n is 2 or 3) at 70⁰C for one to two hours in the presence of a base, for example, potassium carbonate, in dimethylformamide.

Reaction Scheme 5:

wherein A and R₃ and R₄ are respectively the same as defined in

Formula 1 , R_& is a tosyl, mesyl or nosyl group, and n is 2 or 3. The tosyl, mesyl, and nosyl groups represent p-toluenesulfonyl, methanesulfonyl and p-nitrobenzenesulfonyl groups, respectively.

The compounds labeled with a radioactive isotope ¹¹C can be obtained by reacting the compound of Formula 1 with [¹¹C]methoxytriflate ([¹¹C]CH₃OTf) generated from a [¹¹C]carbon dioxide ([¹¹C]CO₂) gas, which is formed in a cyclotron, at 8O⁰C for one minute, followed by purifying the resultant with a high performance liquid chromatography. This reaction can be illustrated in Reaction Scheme 6 below:

Reaction Scheme 6:

wherein A and Ri, R2 and R₄ are respectively the same as defined in Formula ! Meanwhile, the compounds labeled with a radioactive isotope ¹⁸F can be prepared through two steps comprising the reactions as illustrated in Reaction Scheme 7 and a Reaction Scheme 8 below. First, as shown in Reaction Scheme 7, the 5-hydroxyisoindol-1 ,3-dione or 5-hydroxy-2,3- dihydroisoindol-1-one compound is subjected to a nucleophilic substitution with a compound of a general formula ReO(CHb)_nORe (wherein R_& is a tosyl (Ts), mesyl (Ms) or nosyl (Ns) group, and n is 2 or 3) at 70°C for one to two hours in the presence of a base, for example, potassium carbonate, in dimethylformamide, to obtain a reaction intermediate.

Reaction formula:

wherein A and R₃ and R₄ are respectively the same as defined in Formula 1 , R₆ is a tosyl, mesyl, or nosyl group, and n is 2 or 3.

Subsequently, the intermediate obtained by the reaction as shown in Reaction Scheme 7 is reacted with tetrabutyl ammonium ¹⁸fluoride (TBAF¹⁸) at 120⁰C for 10 minutes and then purified with a high performance liquid chromatography, to obtain a compound labeled with a radioactive isotope ¹⁸F. This reaction can be illustrated in Reaction Scheme 8 as follows: Reaction Scheme 8:

wherein A and R₃ and R₄ are respectively the same as defined in Formula 1 , Re is a tosyl, mesyl, or nosyl group, and n is 2 or 3.

The compounds represented by Formulae 4 to 10 below are specific embodiments of the compound of Formula 1 which can be synthesized according to any of the methods as illustrated in Reaction Schemes 1 to 8.

Formula 4:

Formula 5:

Formula 6:

Formula 7:

Formula 8:

Formula 9:

Formula 10:

wherein A, R-i, R2, R3, R₄ and Re are respectively the same as described above, and X is a halogen atom selected from fluorine, chlorine, bromine and iodine.

EXAMPLES

Hereinafter, the present invention will be described in more detail through examples. However, the provided examples are only for illustrative and not intended to limit the scope of the present invention thereto. Example 1 : Preparation of 4-methoxyphthalic anhydride (2a) 1 ,4-Diazabicyclic[2,2,2]octane (336 mg, 3 mmol) was dissolved in anhydrous dichloromethane (3 ml_) and then stirred for five minutes while slowly adding distilled thionyl chloride (0.2 ml_, 3 mmol) at O⁰C. And then,

4-methoxyphthalic acid (200 mg, 1 mmol) was slowly added dropwise at room temperature for 50 minutes. 10% sodium bicarbonate was added to neutralize the reaction solution and the resultant was extracted with dichloromethane.

Organic layers were dried with anhydrous sodium sulfate and filtered, and the filtrate was distilled under reduced pressure, separated and then purified by a column chromatography on silica gel (hexane/ethyl acetate=3:1), to obtain

139 mg of the desired compound (yield 78%).

¹H NMR (300 MHz, CDCI₃) δ 7.92 (d, J=5.6 Hz, 1 H) 7.43 (d, J=2.9, 1 H),

7.36 (dd, J=11.2, 3.0 Hz, 1 H), 3.99 (s, 1 H). 1³C NMR (75 Hz, CDCI₃) δ 166.17, 163.00, 162.43, 134.14, 127.28,

123.22, 123.98, 108.89, 56.44.

Example 2: Preparation of ethyl 2-bromomethyl-4-methoxybenzoate (2b) Ethyl 2-nnethyl-4-methoxybenzoate (583 mg, 3 mmol) and

N-bromosuccinimide (1335 mg, 7.5 mmol) were added to carbon tetrachloride (8 mL), and the resultant was refluxed for six hours. After the reaction was completed, N-bromosuccinimide was filtered out and carbon tetrachloride was distilled under a reduced pressure. The resultant was purified by a column chromatography on silica gel (hexane/ethyl acetate=40:1 ) to obtain 340 mg of the desired compound(yield 48%).

¹H NMR (400 MHz, CDCI₃) δ 7.99 (d, J=2.6 Hz, 1 H), 6.96 (d, J=2.6 Hz, 1 H), 6.85 (dd, J=8.8, 2.6 Hz, 1 H), 4.96 (s, 2H), 4.37 (q, J=7.1 Hz, 2H), 3.86 (s, 3H), 1.40 (t, J=7.1 Hz, 3H).

¹³C NMR (100 MHz, CDCI₃) δ 166.15, 162.46, 141.56, 133.63, 121 .33, 117.03, 113.56, 60.93, 55.50, 31.91 , 14.29.

Example 3: Preparation of 5-methoxy-2-(4-nitrophenyl)isoindol- 1 ,3-dione

1.1 g (6.2 mmol) of anhydrous 4-methoxyphthalic acid compound obtained in Example 1 was dissolved in 10 ml_ of acetic acid, to which 856 mg (6.2 mmol) of 4-nitroaniline was added dropwise, and then the resultant was refluxed for 12 hours. When the reaction was completed, 10% NaHCO₃ solution was added for neutralization. The resultant was extracted with ethyl acetate, and the extract was dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography to obtain 1.7 g (92%) of the desired compound as a pale yellow solid. 1H NMR (300 MHz, DMSOd₆) δ 3.97 (s, 3H), 7.41 (dd, J=2.2, 8.4 Hz,

1 H), 7.53 (d, =2.1 Hz, 1 H), 7.78 (d, J=9.0 Hz, 2H), 7.94 (d, J=8.3 Hz, 1 H), 8.40 (d, J=9.0 Hz, 2H).

Example 4: Preparation of 5-methoxy-2-(4-nitrophenyl)-2,3-dihydro- isoindol-1-one 1.72 g of ethyl 2-bromomethyl-4-methoxybenzoate compound obtained in Example 2 was dissolved in 20 ml_ of acetic acid, to which 870 mg (6.3 mmol) of 4-nitroaniline was added dropwise under a nitrogen atmosphere, and the resultant was refluxed for five hours. When the reaction was completed, 10% NaHCO₃ solution was added for neutralization. The resultant was extracted with ethyl acetate, and the extract was dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography to obtain 900 g (50%) of the desired compound as a yellow solid. 1H NMR (300 MHz, DMSO-d₆) δ 3.89 (s, 3H), 5.06 (s, 2H), 7.11 (d,

J=8.5 Hz, 1 H), 7.23 (s, 1 H), 7.75 (d, J=8.5 Hz, 1 H), 7.16 (d, J=9.4 Hz, 2H), 8.32 (d, J=9.4 Hz, 2H).

Example 5: Preparation of 5-methoxy-2-(4-aminophenyl)-2,3-dihydro- isoindol-1-one (1b-22)

25 ml_ of ethanol was added dropwise to 531 mg (1.86 mmol) of 5-methoxy-2-(4-nitrophenyl)-2,3-dihydroisoindol-1-one compound obtained in Example 4, to which 2.1 g (9.3 mmol) of tin chloride was added, and the resultant was then refluxed for 24 hours. When the reaction was completed, 1 N sodium hydroxide solution was added to adjust pH of the solution to 10. The resultant was distilled under a reduced pressure to remove ethanol, and the residue was filtered using ethyl acetate, and extracted with water and ethyl acetate. Organic layers were dried with anhydrous sodium sulfate and then filtered to obtain 450 mg (yield 95%) of the desired compound as a yellow solid. ¹H NMR (300 MHz, DMSO-CZ₆) δ 3.85 (s, 3H), 5.02 (s, 2H), 5.33 (s, NH₂), 6.60 (d, J=8.6 Hz, 2H), 7.05 (d, J=8.4 Hz, 1 H), 7.16 (s, 1 H), 7.43 (d, J=8.6 Hz, 2H), 7.63 (d, J=8.4 Hz, 1 H).

Example 6: Preparation of 5-hydroxy-2-(4-aminophenyl)-2,3-dihydro- isoindol-1-one (1b-24)

50.8 mg (0.2 mmol) of 5-methoxy-2-(4-aminophenyl)-2,3- dihydroisoindol-1-one compound obtained in Example 5 was dissolved in 8 ml_ of anhydrous dichloromethane, to which 2 ml_ (2 mmol) of boron tribromide (BBr₃) was added under a nitrogen atmosphere at O⁰C, and the resultant was then refluxed for 12 hours. When the reaction was completed, the resultant was neutralized with 1 N sodium hydroxide, extracted with ethyl acetate, and then, the extract was dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography to obtain 28 mg (58%) of the desired compound as a beige colored solid.

¹H NMR (300 MHz, DMSOd₆) δ 4.75 (s, 2H), 5.00 (s, NH₂), 6.59 (d, J=8.1 Hz, 2H), 6.86 (d, J=8.5 Hz, 1H), 6.91 (s, 1H), 7.42 (d, J=8.2 Hz, 2H), 7.51 (d, J=8.7 Hz, 1 H), 10.39 (bs, OH).

Example 7: Preparation of 5-hydroxy-2-(4-methylaminophenyl)-2,3- dihydroisoindol-1-one (1 b-25)

50.8 mg (0.2 mmol) of

5-hydroxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1 -one compound obtained in Example 6 was dissolved in 8 ml_ of anhydrous dichloromethane, to which 2 ml_ (2 mmol) of boron tribromide (BBr₃) was added under a nitrogen atmosphere at 0⁰C, and the resultant was then refluxed for 12 hours. When the reaction was completed, the resultant was neutralized with 1N sodium hydroxide, extracted with ethyl acetate, and then, the extract was dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography to obtain 28 mg (58%) of the desired compound as a beige colored solid.

¹H NMR (300 MHz, DMSOd₆) δ 4.75 (s, 2H), 5.00 (s, NH₂), 6.59 (d, J=8.1 Hz, 2H), 6.86 (d, J=8.5 Hz, 1H), 6.91 (s, 1H), 7.42 (d, J=8.2 Hz, 2H), 7.51 (d, J=8.7 Hz, 1 H), 10.39 (bs, OH).

Example 8: Preparation of 5-methoxy-2-(4-dimethylaminophenyl)-2,3- dihydroisoindol-1-one (1b-1 ) Ethyl 2-bromomethyl-4-methoxybenzoate compound (273 mg, 1 mmol) obtained in Example 2 and N,N-dimethylbenzene-1 ,4-diamine (136 mg, ammol) was refluxed in acetic acid (1 ml_) at 12O⁰C for four hours. 10% sodium hydrogen carbonate was added to neutralize the reaction solution, which was then extracted using water and dichloromethane. Organic layers were dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography on silica gel (hexane/ethyl acetate=3:1 ) to obtain 78 mg of the desired compound (yield 9%).

¹H NMR (300 MHz, CDCI₃) δ 7.82 (d, J=11.1 Hz, 1 H), 7.63 (d, J=12.1 Hz, 2H), 7.00 (dd, J=11.2, 2.8 Hz, 1 H), 6.97 (s, 1 H), 6.79 (d, J=12.2 Hz, 2H), 4.74 (s, 2H), 3.89 (s, 3H), 2.95 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.02, 162.84, 147.99, 142.51 , 129.48, 126.20, 125.27, 121.50, 114.78, 113.13, 107.39, 55.67. 51.09, 40.88.

Example 9: Preparation of 5-(2-fluoroethoxy)-2-(4-aminophenyl)-2,3- dihydroisoindol-1-one (1b-26)

7.2 mg (30 mmol) of 5-hydroxy-2-(4-aminophenyl)-2,3-dihydro- isoindol-1-one compound obtained in Example 6 was dissolved in 3 mL of anhydrous dimethylformamide, to which 6.2 mg (45 mmol) of potassium carbonate (K₂CO₃) and 9.8 mg (45 mmol) of 1-fluoro-2-tosyloxyethane were added under a nitrogen atmosphere at O⁰C, and the resultant was then reacted at 7O⁰C for two hours. When the reaction was completed, the resultant was extracted using an excess amount of water and ethyl acetate, and organic layers were dried with anhydrous sodium sulfate and filtered. The solvent was distilled off under a reduced pressure, and the residue was purified by a column chromatography to obtain 5 mg (59%) of the desired compound as a beige colored solid.

¹H NMR (300 MHz, DMSO-c/₆) δ 4.30 (m, 1 H), 4.39 (m, 1 H), 4.69 (m, 1 H), 4.82 (s, 2H), 4.85 (m, 1 H), 5.03 (s, NH₂), 6.50 (d, J=8.6 Hz, 2H), 7.09 (d, J=8.1 Hz, 1 H), 7.19 (s, 1 H), 7.44 (d, J=7.9 Hz, 2H), 7.63 (d, J=8.4 Hz, 1 H).

Example 10: Preparation of 5-methoxy-2-(4-¹¹methylaminophenyl)-2,3- dihydroisoindol-1-one (1b-32) [¹¹C]CO₂ gas was generated from nitrogen gas in a cyclotron and then transferred from the target of the cyclotron to a [¹¹C]CHaOTf synthesizing device using helium gas. The [¹¹C]CO₂ gas was converted into [¹¹C]CH₄ with an oven in the [¹¹C]CH₃OTf synthesizing device, and then [¹¹C]CH₄ was then reacted with I₂ gas at 720⁰C to generate [¹¹C]CH₃I. The generated [¹¹C]CH₃I was reacted with silver triflate at 200⁰C to generate [¹¹C]CH₃OTf. 3.0 mg of 5-methoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1 -one compound obtained in Example 5 was dissolved in 400 μl_ of methyl ethyl ketone, and then, the generated [¹¹C]CH₃OTf was collected at 125°C. After the collection was completed, the reaction was carried out for one minute at 80⁰C, and then purified through a high performance liquid chromatography to obtain the pure title compound.

For the purification using the high performance liquid chromatography, a high performance liquid chromatography pump of the [¹¹C]CH₃OTf synthesizing device, a UV detecting device, a sodium iodide (NaI) pin detector system were used. In injecting a sample, an automatic injecting device in an automatic synthetic device was used, and elution was performed at 4.0 mL/min with 50 mM TEAP:CH₃CN = 8:2. As a column, prodigy ODS-prep 10 mm (250 x 10 mm) was used.

Example 11 : Preparation of 5-(2-tosyloxyethoxy)-2-(4-dimethylamino- phenyl)-2,3-dihydroisoindol-1-one

Anhydrous dimethylformamide (3 ml_) was added to 5-hydroxy-2-(4- aminophenyl)-2,3-dihydroisoindol-1-one compound (48 mg, 0.18 mmol) obtained in Example 6, and the resultant was stirred under a nitrogen atmosphere. Potassium carbonate (37 mg, 0.27 mmol) was added thereto, to which 1 ,2-bis(tosyloxy)ethane (100 mg, 0.27 mmol) was added, and the resultant was heated at 80⁰C. When the reaction was completed after two hours, the resultant was cooled and then extracted with ethyl acetate and water. Organic layers were dried with anhydrous sodium sulfate and filtered, and the solvent was distilled off under a reduced pressure. The residue was purified by a column chromatography to obtain the desired compound (33 mg, yield 40%). 1H NMR (300 MHz, DMSO-d₆) δ 7.80 (d, J=8.1 Hz, 2H), 7.62 (m, 3H),

7.65 (d, J=8.1 Hz, 2H), 7.07 (s, 1 H), 6.95 (d, J=8.3 Hz, 1 H), 6.78 (d, J=9.1 Hz, 2H), 4.84 (s, 2H), 4.37 (d, J=3.1 Hz, 2H), 4.28 (d, J=3.1 Hz, 2H), 2.89 (s, 6H), 2.40 (s, 3H).

Example 12: Preparation of 5-(2-¹⁸ fluoroethoxy)-2-(4-dimethylamino- phenyl)-2,3-dihydroisoindol-1-one (1b-33)

Fluorine-18 (319.2 MBq) in water (0.5 mL) was maintained at a quaternary ammonium salt-supported polymer cartridge (Chromafix^®), and then eluted with 620 mL of mixed solution of 300 mL of water and 300 mL of acetonitrile containing an aqueous solution in which tetrabutylammonium salt and bicarbonate ion were dissolved. Azeotropic distillation was performed with acetonitrile (3 x 500 mL) at 100⁰C while blowing nitrogen gas so as to completely remove moisture from this solution. The obtained tetrabutylammonium ¹⁸fluoride (TBAF¹⁸) was dissolved in 200 mL of acetonitrile and 500 ml_ of t-amyl alcohol, to which 4.0 mg (8.57 mmol) of 5-(2-tosyloxyethoxy)-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one obtained in Example 11 was added. The reaction mixture was heated at 12O⁰C for ten minutes to obtain 5-(2-¹⁸fluoroethoxy)-2-(4-dimethyl- aminophenyl)-2,3-dihydroisoindol-1-one compound. The reaction yield (39.07%) was checked by a thin layer chromatography. The reaction solvent was removed by blowing nitrogen gas at 120⁰C, and the residue was dissolved in 1 ml. of acetonitrile and then purified by a high performance liquid chromatography. The resultant was injected together with 5-(2-fluoroethoxy)-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one compound obtained in Example 9, so as to confirm that the desired compound was synthesized. A peak having identical retention time at 9.5 minutes in UV and radioactive chromatogram were obtained.

For the purification using the high performance liquid chromatography, a high performance liquid chromatography pump of, a UV detector (TSP, USA), a sodium iodide (NaI) pin detector system (Bioscan, Washington DC, USA) were used. In injecting the sample, 2 mL of a high performance liquid chromatography loop and a manual injector were used. Elution was performed at 4.0 mL/min with water:acetonitrile=60:40. As a column, prodigy ODS-prep 10 mm (250 x 10 mm) was used.

Examples 13 to 65

The compounds of Examples 13 to 65 below were prepared in a similar manner to that of Examples 1 to 9. The structure identification data of those compounds are provided below.

Example 13: 5-dimethylamino-2-(4-dimethylaminophenyl)isoindol- 1 ,3-dione (1a-1) 1H NMR (400 MHz, CDCI₃) δ 7.73 (d, J=8.5 Hz, 1 H), 7.24 (d, J=8.6 Hz,

2H), 7.15 (s, 1 H), 6.84 (d, J=9.4 Hz, 1 H) 6.80 (d, J=8.6 Hz, 2H), 3.13 (s, 6H), 2.98 (s, 6H).

¹³C NMR (100 MHz, CDCI₃) δ 168.75, 168.37, 154.46, 150.06, 134.65, 127.62, 125.06, 120.86, 117.54, 114.86, 112.60, 105.85, 40.62, 40.52.

Example 14: 5-methoxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-2)

¹H NMR (400 MHz, CDCI₃) δ 7.83 (d, J=8.3 Hz, 1 H), 7.41 (d, J=2.2 Hz 1 H), 7.27-7.19 (m, 3H), 6.80 (dd, J=7.0, 2.0 Hz), 3.95 (s, 3H), 3.00 (s, 6H). 1³C NMR (100 MHz, CDCI₃) δ 167.74, 164.79, 150.21 , 134.62, 127.54,

125.23, 123.88, 120.27, 120.11 , 112.52, 108.06, 56.13, 40.55.

Example 15: 5-hydroxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-3) 1H NMR (300 MHz, DMSOd₆) δ 11.0 (s, 1 H), 7.75 (d, J=10.7 Hz, 1 H),

7.18-7.15 (m, 4H), 6.80 (d, J=11.9 Hz, 2H), 2.94 (s, 6H).

¹³C NMR (75 MHz, DMSO-d₆) δ 167.67, 163.81 , 134.80, 128.54, 125.92, 122.17, 121.04, 113.57, 110.27, 41.25.

Example 16: 5-methyl-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-4)

¹H NMR (300 MHz, CDCI₃) δ 7.82 (d, J=I. Q Hz, 1 H)₁ 7.75 (s, 1 H), 7.56 (d, J=10.2 Hz, 1 H), 7.27-7.25 (m, 2H), 6.85 (bs, 2H), 3.01 (s, 6H), 2.55 (s, 3H). 1³C NMR (75 MHz, CDCI₃) δ 168.11 , 168.00, 150.16, 145.41 , 134.69,

132.35, 129.38, 127.52, 124.03, 123.44, 120.31 , 112.55, 40.58, 22.05.

Example 17: 2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-5) 1H NMR (300 MHz, CDCI₃) δ 7.97-7.92 (m, 2H), 7.80-7.76 (m, 2H), 7.26 (d, J=7.1 Hz, 2H), 6.83 (d, J=8.2 Hz, 2H), 3.02 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.95, 150.20, 134.16, 131.97, 127.54, 123.54, 120.10, 112.56, 40.60.

Example 18: 5-fluoro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-6)

¹H NMR (300 MHz, CDCI₃) δ 7.94 (dd, J=8.2, 4.5 Hz, 1 H), 7.62-7.59 (m, 1 H), 7.47-7.40 (m, 1 H), 7.24 (d, J=8.9 Hz, 2H), 6.82 (d, J=8.8 Hz, 2H), 3.01 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 168.19, 166.86, 166.54, 166.51 , 164.79, 150.25, 134.88, 134.76, 127.76, 127.72, 127.43, 125.98, 125.85, 121.31 , 121.00, 119.93, 112.53, 111.45, 111.12, 40.55.

Example 19: 5-chloro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione (1a-7) 1H NMR (300 MHz, CDCI₃) δ 7.90 (d, J=1.1 Hz, 1 H), 7.86 (d, J=6.0 Hz, 1H), 7.73 (dd, J=6.0, 1.3 Hz, 1H), 7.24 (d, J=6.7 Hz, 2H), 6.84 (d, J=5.0 Hz, 2H), 3.00 (s, 6H).

¹³C NMR (75 MHz, DMSO-d₆) δ 167.00, 166.67, 150.34, 140.80, 134.19, 133.65, 130.03, 127.42, 124.80, 123.97, 119.65, 112.44, 40.51.

Example 20: 5-bromo-2-(4-dimethylaminophenyl)isoindol-1,3-dione (1a-8)

¹H NMR (300 MHz, CDCI₃) δ 8.07 (d, J=1.3 Hz, 1H), 7.91 (dd, J=7.9, 1.6 Hz, 1H), 7.80 (d, J=7.9 Hz, 1H), 7.25-7.22 (m, 2H), 6.81 (d, J=9.0 Hz, 2H), 3.01 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.09, 166.55, 150.35, 137.13, 133.63, 130.52, 129.00, 127.40, 126.86, 124.90, 119.72, 112.45,40.49.

Example 21 : 5-dimethylamino-2-(4-dimethylamino-3-methoxyphenyl)- isoindol-1,3-dione (1a-9)

¹H NMR (300 MHz, CDCI₃) δ 7.74 (d, J=8.5 Hz, 1H), 7.15 (d, J=2.3 Hz, 1H), 7.02 (d, J= 8.4 Hz, 1H), 6.95 (dd, J=8.4, 2.1 Hz, 1H), 6.88 (d. J=2.1 Hz, 1H), 6.85 (dd, J=8.6, 2.4 Hz, 1H), 3.90 (s, 3H), 3.16 (s, 6H), 2.82 (s, 6H).

¹³C NMR (75 MHz, DMSO-d₆) δ 168.51, 168.08, 154.59, 152.45, 134.50, 125.26, 119.18, 118.22, 117.26, 115.06, 109.96, 105.90, 55.57, 43.31, 40.56.

Example 22: 5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)- isoindol-1,3-dione(1a-10) 1H NMR (300 MHz, CDCI₃) δ 7.86 (d, J=8.3 Hz, 1H), 7.43 (d, J=2.2 Hz, 1 H), 7.23 (dd, J= 8.3, 2.3 Hz, 1 H), 7.03 (d, J=8.4 Hz, 1 H), 3.96 (dd, J= 8.4, 2.1 Hz, 1 H), 6.88 (d, J=2.0 Hz, 1 H), 3.97 (s, 3H), 3.91 (s, 3H), 2.83 (s, 6H).

¹³C NMR (75 MHz, DMSO-d₆) δ 167.50, 165.02, 152.51 , 142.26, 134.51 , 125.92, 125.47, 123.73, 120.38, 119.18, 118.25, 109.90, 108.25, 56.23, 55.64, 43.28.

Example 23: 5-methyl-2-(4-dimethylamino-3-methoxyphenyl)isoindol- 1 ,3-dione (1a-11 )

¹H NMR (300 MHz, CDCI₃) δ 7.82 (d, J=7.6 Hz, 1H), 7.57 (d, J=7.1 Hz, 1 H), 7.02 (d, J=8.4 Hz, 1H), 6.95 (dd, J=8.4, 2.1 Hz, 1 H), 6.87 (d, J=2.0 Hz, 1 H), 3.90 (s, 3H), 2.82 (s, 6H), 2.55 (s, 3H).

¹³C NMR (75 MHz, DMSO-d₆) δ 167.81 , 167.70, 152.43, 145.65, 134.87, 132.22, 129.25, 124.16, 123.57, 120.42, 119.12, 118.11 , 109.88, 55.57, 43.20, 22.05.

Example 24: 2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione (1a-12).

¹H NMR (300 MHz, CDCI₃) δ 7.97-7.80 (m, 2H), 7.78-7.77 (m, 2H), 7.03 (d, J=8.4 Hz, 1 H), 6.96 (dd, J=8.4, 2.2 Hz, 1 H), 6.88 (d, J=2.1 Hz, 1 H), 3.90 (s, 3H), 2.83 (s, 6H).

¹³C NMR (75 MHz, DMSOd₆) δ 167.64, 152.45, 142.46, 134.32, 131.84, 125.67, 123.67, 119.14, 118.11 , 109.86, 55.58, 43.19.

Example 25: 2-(4-aminostilbene)-5-(3-fluoropropyloxy)benzoxazole (1a-13) ¹H NMR (300 MHz, CDCI₃) δ 7.98 (dd, J=8.2, 4.5 Hz, 1 H), 7.63 (dd, J=I Λ, 2.1 Hz, 1 H), 7.46 (dt, J=8.5, 2.3 Hz, 1 H), 6.98 (dd, J=8.4, 2.1 Hz, 1 H), 6.91 (d, J=1.7 Hz, 1 H), 3.92 (s, 3H), 2.88 (s, 6H).

¹³C NMR (75 MHz, DMSOd₆) δ 166.60, 166.05, 152.45, 142.63, 127.56, 126.14, 121.53, 121.29, 119.05, 118.13, 111.58, 111.33, 109.66, 55.59, 43.17.

Example 26: 5-chloro-2-(4-dimethylamino-3-methoxyphenyl)isoindol- 1 ,3-dione (1a-14) 1H NMR (300 MHz, CDCI₃) δ 7.93 (d, J=1.4 Hz, 1 H), 7.89 (d, J=7.9 Hz,

1 H), 7.76 (dd, J=8.0, 1.8 Hz, 1 H), 7.03 (d, J=8.4 Hz, 1 H), 6.96 (dd, J=8.4, 2.2 Hz, 1 H), 6.88 (d, J=2A Hz, 1 H), 3.91 (s, 3H), 2.84 (s, 6h).

¹³C NMR (75 MHz, DMSO-d₆) δ 166.67, 166.17, 152.48, 141.10, 134.45, 133.46, 129.84, 124.95, 124.12, 119.08, 109.81 , 55.65, 43.25.

Example 27: 5-bromo-2-(4-dimethylamino-3-methoxyphenyl)isoindol- 1 ,3-dione (1a-15)

¹H NMR (300 MHz, CDCI₃) δ 8.09 (d, J=I .2 Hz, 1 H), 7.93 (dd, J=7.9, 7.9 Hz, 1 H), 7.82 (dd, J=7.9, 0.3 Hz, 1H), 7.03 (d, J=8.4 Hz, 1 H), 6.95 (dd, J= 8.4, 2.2 Hz, 1 H), 6.87 (d, J=2.2 Hz, 1 H).

¹³C NMR (75 MHz, DMSO-d₆) δ 166.80, 166.26, 152.45, 137.36, 133.45, 130.42, 129.25, 127.01 , 125.03, 119.06, 118.16, 109.77, 55.61 , 43.15.

MS m/z 375 (M⁺). Example 28: 2-(3-dimethylamino-4-methoxyphenyl)isoindol-1 ,3-dione (1a-16)

¹H NMR (300 MHz, CDCI₃) δ 7.97-7.94 (m, 2H), 7.81-7.78 (m, 2H), 7.02 (dd, J=8.6, 2.2 Hz, 1 H), 6.95 (d, J=2.1 Hz, 1 H), 3.94 (s, 3H), 2.83 (s, 6H).

¹³C NMR (75 MHz, DMSOd₆) δ 167.71 , 152.16, 143.02, 134.26, 131.90, 124.42, 123.62, 120.65, 117.03, 111.14, 55.63, 43.15.

Example 29: 2-(3-dimethylaminophenyl)isoindol-1 ,3-dione (1a-17). 1H NMR (300 MHz, CDCI₃) δ 7.97-7.92 (m, 2H), 7.80-7.75 (m, 2H)

7.34 (t, J=8.0 Hz, 1 H), 6.78-6.70 (m, 3H), 2.98 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.52, 151.16, 134.24, 132.43, 131.94, 129.56, 123.64, 114.61 , 114.40, 110.78, 40.46.

Example 30: 5-methoxy-2-(4-aminophenyl)isoindol-1 ,3-dione (1a-18)

¹H NMR (300 MHz, DMSO-Cf₆) δ 3.94 (s, 3H), 5.33 (s, NH₂), 6.62 (d, J=8.6 Hz, 2H), 6.98 (d, J=8.6 Hz, 2H), 7.35 (dd, J=2.2, 8.3 Hz, 1 H), 7.43 (d, J=2.1 Hz, 1 H), 7.84 (d, J=8.3 Hz, 1 H).

Example 31 : 5-methoxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione

(1a-19)

¹H NMR (300 MHz, DMSO-c/₆) δ 2.69 (d, J=4.7 Hz, 3H), 3.69 (s, 3H)

5.91 (d, J=4.5 Hz, NH), 6.60 (d, J=8.4 Hz, 2H), 7.07 (d, J=8.3 Hz, 2H), 7.35 (d,

J=8.2 Hz, 1 H), 7.43 (s, 1 H), 7.84 (d, J=8.2 Hz, 1 H). Example 32: 5-hydroxy-2-(4-aminophenyl)isoindol-1 ,3-dione (1a-20) 1H NMR (300 MHz, DMSO-cfe) δ 5.31 (s, NH₂), 6.61 (d, J=8.6 Hz, 2H), 6.96 (d, J=8.5 Hz, 2H), 7.13 (m, 2H), 7.72 (d, J=8.0 Hz, 1 H), 11.24 (bs, OH).

Example 33: 5-hydroxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione

(1a-21 ).

¹H NMR (300 MHz, DMSO-d₆) δ 2.69 (d, J=4.8 Hz, 3H), 5.90 (m, NH), 6.59 (d, J=8.6 Hz, 2H), 7.05 (d, J=8.6 Hz, 2H), 7.15 (m, 2H), 7.74 (d, J=7.9 Hz, 1 H), 10.97 (bs, OH).

Example 34: 5-(2-fluoroethoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione (1a-22).

¹H NMR (300 MHz, DMSO-d₆) δ 4.42 (m, 1 H), 4.51 (m, 1H), 4.71 (m, 1 H), 4.87 (m, 1 H), 5.33 (s, NH₂), 6.62 (d, J=8.6 Hz, 2H), 6.98 (d, J=8.5 Hz, 2H), 7.39 (dd, J=2.0, 8.3 Hz, 1 H), 7.47 (d, J=2.0 Hz, 1 H), 7.84 (d, J=8.3 Hz, 1 H).

Example 35: 5-(2-fluoroethoxy)-2-(4-methylaminophenyl)isoindol- 1 ,3-dione (1a-23) 1H NMR (300 MHz, DMSO-d₆) δ 2.70 (d, J=4.9 Hz, 3H), 4.42 (m, 1 H),

4.52 (m, 1 H), 4.71 (m, 1 H), 4.87 (m, 1 H), 5.91 (s, NH), 6.60 (d, J=8.7 Hz, 2H), 7.07 (d, J=8.5 Hz, 2H), 7.39 (dd, J=2.1 , 8.3 Hz, 1 H), 7.48 (d, J=2.0 Hz, 1 H), 7.85 (d, J=8.3 Hz, 1 H).

Example 36: 5-(2-fluoroethoxy)-2-(4-dimethylaminophenyl)isoindol- 1,3-dione(1a-24).

¹H NMR (300 MHz, DMSO-^₆) δ 2.94 (s, 6H), 4.43 (m, 1H), 4.52 (m, 1H), 4.71 (m, 1H), 4.88 (m, 1H), 6.79 (d, J=8.9 Hz, 2H), 7.18 (d, J=8.9 Hz, 2H), 7.40 (dd, J=2.1, 8.3 Hz, 1H), 7.49 (d, J=2.0 Hz, 1H), 7.86 (d, J=8.3 Hz, 1H).

Example 37: 5-(3-fluoropropoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione (1a-25).

¹H NMR (300 MHz, DMSO-cfe) δ 2.11 (t, J=5.9 Hz, 1H), 2.19 (t, J=5.9 Hz, 1H), 4.28 (t, J=6.0 Hz, 2H), 4.55 (t, J=5.6 Hz, 1H), 4.70 (t, J=5.7 Hz, 1H), 5.31 (s, 2H), 6.62 (d, J=8.5 Hz, 2H), 6.98 (d, J=8.4 Hz, 2H), 7.36 (d, J=8.2 Hz, 1H), 7.43 (s, 1H), 7.83 (d, J=8.3 Hz, 1H).

Example 38: 5-(3-fluoropropoxy)-2-(4-methylaminophenyl)isoindol- 1,3-dione(1a-26).

¹H NMR (300 MHz, DMSO-d₆) δ 2.11 (t, J=5.7, 6.1 Hz, 1H), 2.20 (t,

J=6.0, 6.1 Hz, 1H), 2.70 (d, J=5.0 Hz, 3H), 4.28 (t, J=6.2 Hz, 2H), 4.55 (t,

J=5.9 Hz, 1H), 4.71 (t, J=5.9 Hz, 1H), 5.91 (m, NH), 6.60 (d, J=8.8 Hz, 2H),

7.07 (d, J=8.7 Hz, 2H), 7.36 (dd, J=2.2, 8.3 Hz, 1H), 7.44 (d, J=2.1 Hz, 1H), 7.84 (d, >8.3Hz, 1H).

Example 39: 5-(3-fluoropropoxy)-2-(4-dimethylaminophenyl)isoindol- 1,3-dione(1a-27).

¹H NMR (300 MHz, DMSO-d₆) δ 2.11 (t, J=5.7, 6.2 Hz, 1H), 2.20 (t, J=5.7, 6.1 Hz, 1H), 2.94 (s, 6H), 4.28 (t, J=6.0, 6.2 Hz, 2H), 4.55 (t, J=5.6 Hz, 1 H)₁ 4.71 (t, J=5.5, 5.7 Hz, 1 H), 6.79 (d, J=8.8 Hz, 2H), 7.17 (d, J=8.6 Hz, 2H), 7.37 (d, J=8.3 Hz, 1 H), 7.45 (s, 1 H), 7.84 (d, J=8.3 Hz, 1 H).

Example 40: 5-hydroxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol- 1-one (1 b-2)

¹H NMR (400 MHz, CDCI₃) δ 10.20, 7.60 (d, J=8.9 Hz, 2H), 7.52 (d, J=8.3 Hz, 1 H), 6.92 (s, 1H), 6.86 (d, J=8.2 Hz, 1H), 6.76 (s, 2H), 4.78 (s, 2H), 2.86 (s, 6H).

¹³C NMR (100 MHz, CDCI₃) δ 166.53, 161.44, 147.67, 143.75, 130.07, 124.94, 124.31 , 121.21 , 116.22, 113.21 , 109.73, 50.78, 40.91.

Example 41 : 5-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1 b-3)

¹H NMR (300 MHz, CDCI₃) δ 7.78 (d, J=8.2 Hz, 1 H), 7.66-7.64 (m, 2H), 7.36-7.28 (m, 2H), 6.80 (d, J=8.9 Hz, 2H), 4.75 (s, 2H), 2.96 (s, 6H), 2.48 (s, 3H).

¹³C NMR (75 MHz, CDCI₃) δ 167.23, 148.05, 142.18, 140.65, 131.02, 129.23, 123.70, 122.97, 121.59, 113.10, 51.16, 40.89.

Example 42: 2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one

(1b-4).

¹H NMR (300 MHz, CDCI₃) δ 7.94-7.91 (m, 1 H), 7.69 (d, J=9.1 Hz, 2H),

7.58-7.56 (m, 1 H), 7.53-7.49 (m, 2H), 6.85 (d, J=7.8 Hz, 2H), 4.82 (s, 2H),

2.98 (s, 6H). 1³C NMR (75 MHz, CDCI₃) δ 167.08, 148.16, 140.27, 133.58, 131.50, 129.19, 128.19, 123.90, 122.48, 121.66, 113.04, 51.33, 40.81.

Example 43: 5-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1b-5) 1H NMR (300 MHz, CDCI₃) δ 7.88 (dd, J=9.1 , 5.1 Hz, 1 H), 7.61 (dd,

J=6.9, 2.2 Hz, 2H), 7.21-7.16 (m, 2H), 6.78 (dd, J=6.9, 2.2 Hz, 2H), 4.78 (s, 2H), 2.96 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 166.83, 166.09, 163.50, 148.24, 142.64, 142.50, 129.59, 128.88, 126.00, 125.87, 121.70, 116.20, 115.89, 113.00, 110.08, 109.76, 51.01 , 50.98, 40.80.

Example 44: 5-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1 b-6)

¹H NMR (300 MHz, CDCI₃) δ 7.82 (d, J=8.6 Hz, 1H), 7.63 (d, J=9.0 Hz, 2H), 7.46 (m, 2H), 6.79 (d, J=8.6 Hz, 2H), 4.75 (s, 2H), 2.96 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 166.01 , 148.26, 141.78, 137.80, 132.08, 128.86, 128.72, 125.10, 122.94, 121.66, 112.94, 50.87, 40.76.

Example 45: 5-bromo-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1b-7)

¹H NMR (400 MHz, CDCI₃) δ 7.87 (s, 1 H), 7.61 (d, J=8.9 Hz, 2H), 7.52 (dd, J=8.0, 1.2 Hz, 1 H), 7.41 (d, J=8.0 Hz, 1 H), 6.78 (d, J=8.9 Hz, 2H), 4.77 (s, 2H), 2.96 (s, 6H).

¹³C NMR (100 MHz, CDCI₃) δ 165.74, 148.33, 138.36, 135.33, 134.54, 131.68, 128.68, 124.04, 123.78, 121.74, 112.94, 51.05, 40.76. Example 46: 6-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1b-8)

¹H NMR (300 MHz, CDCI₃) δ 7.71-7.67 (m, 3H), 7.37 (s, 2H), 6.88 (bs, 2H), 4.75 (s, 2H), 2.97 (s, 6H), 2.46 (s, 3H).

Example 47: 6-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1 b-9)

¹H NMR (400 MHz, CDCI₃) δ 7.67-7.63 (m, 2H), 7.58 (dd, J=7.7, 2.4 Hz, 1 H), 7.49-7.44 (m, 1 H), 7.31-7.24 (m, 1H), 6.83-6.79 (m, 2H), 4.79 (s, 2H), 2.98 (s, 6H).

¹³C NMR (100 MHz, CDCI₃) δ 166.06, 164.22, 161.77, 148.31 , 135.68, 128.79, 124.05, 123.96, 121.74, 119.20, 118.96, 112.95, 110.75, 110.51 , 50.98, 40.78.

Example 48: 6-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol- 1-one (1 b-10)

^*312¹H NMR (300 MHz, CDCI₃) δ 7.88 (d, J=I .7 Hz, 1 H), 7.65 (m, 2H), 7.53 (dd, J=8.0, 1.7 Hz, 1 H), 7.43 (d, J=8.1 Hz, 1 H), 6.81-6.78 (m, 2H), 4.78 (s, 2H), 2.98, (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 165.71 , 148.29, 138.36, 135.33, 134.52, 131.67, 128.70, 124.02, 123.78, 121.69, 112.94, 51.02, 40.76.

Example 49: 5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1 b-11 ) ¹H NMR (300 MHz, CDCI₃) δ 7.89 (s, 1 H), 7.81 (d, J=8.3 Hz, 1 H), 7.02 (d, J=8.5 Hz, 1 H), 6.99 (s, 1H), 3.96 (s, 2H), 4.78 (s, 2H), 3.96 (s, 3H), 3.90 (s, 3H), 2.80 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.33, 163.23, 152.70, 142.34, 125.84, 125.38, 118.79, 115.10, 110.49, 107.36, 104.09, 55.70, 55.57, 50.77, 43.61.

Example 50: 5-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1 b-12)

¹H NMR (300 MHz, CDCI₃) δ 8.00 (d, J=8.1 Hz, 1 H), 7.79 (s, 1 H), 7.74 (d, J=8.4 Hz, 2H), 6.95 (d, J=8.2 Hz, 1 H), 6.87-6.85 (m, 2H), 4.78 (s, 2H), 3.94 (s, 3H), 2.83 (s, 6H), 2.67 (s, 3H).

Example 51 : 2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydro- isoindol-1-one (1 b-13) 1H NMR (300 MHz, CDCI₃) δ 7.93-7.90 (m, 2H), 7.62-7.58 (m, 1 H),

7.53-7.52 (m, 2H), 7.04-6.96 (m, 2H), 4.85 (s, 2H), 3.97 (s, 3H), 2.81 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 167.41 , 152.64, 140.08, 134.79, 466.32, 131.94, 130.81 , 128.38, 123.96, 122.56, 118.10, 110.98, 104.24, 55.53, 51.13, 43.45.

Example 52: 5-fluoro-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1b-14)

¹H NMR (300 MHz, CDCI₃) δ 7.91-7.87 (m, 1 H), 7.83 (s, 1 H), 7.26-7.18 (m, 2H), 6.97 (s, 2H), 4.83 (s, 2H), 3.96 (s, 3H), 2.80 (s, 6H). 1³C NMR (75 MHz, CDCI₃) δ 167.04, 166.33, 152.65, 142.47, 142.39, 139.54, 134.39, 129.40, 126.13, 126.00, 117.98, 116.44, 116.13, 110.98, 110.16, 109.84, 104.16, 55.53, 50.79, 43.42.

Example 53: 5-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1b-15)

¹H NMR (300 MHz, CDCI₃) δ 7.82 (m, 2H), 7.48 (m, 2H), 6.96 (m, 2H), 4..81 (s, 2H), 3.95 (s, 3H), 2.80 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 166.27, 152.63, 14.60, 139.65, 138.24, 134.23, 131.88, 129.05, 125.18, 123.01 , 117.96, 111.05, 104.22, 55.54, 50.67, 43.39.

Example 54: 5-bromo-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1 b-16)

¹H NMR (300 MHz, CDCI₃) δ 7.86 (s, 2H), 7.23 (m, 2H), 7.00 (m, 2H), 4.82 (s, 2H), 3.96 (s, 3H), 2.82 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 166.36, 152.67, 129.34, 126.02, 118.30, 116.41 , 116.19, 110.92, 110.13, 109.89, 104.21 , 55.56, 50.74, 43.48.

Example 55: 6-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1b-17)

¹H NMR (300 MHz, CDCI₃) δ 7.90 (s, 1 H), 7.69 (s, 1 H), 7.38 (s, 2H), 6.70 (s, 2H), 4.78 (s, 2H), 3.96 (s, 3H), 2.81 (s, 6H), 2.46 (s, 3H).

¹³C NMR (75 MHz, CDCI₃) δ 167.54, 152.64, 138.41 , 137.30, 135.16, 133.44, 132.99, 124.12, 122.27, 118.25, 110.86, 104.19, 55.54, 50.90, 43.50, 21.40. MS m/z 296 (M⁺).

Example 56: 6-fluoro-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1 b-18) 1H NMR (300 MHz, CDCI₃) δ 7.87 (s, 1H), 7.57 (dd, J=7.6, 2.3 Hz, 1H),

7.51-7.47 (m, 1 H), 7.31 (dd, J=8.4, 2.3 Hz, 1 H), 7.01 (s, 2H), 4.83 (s, 2H), 3.97 (s, 3H), 2.83 (s, 6H).

¹³C NMR (75 MHz, CDCI₃) δ 161.41 , 152.66, 135.44, 124.17, 124.06, 119.72, 119.40, 111.06, 110.83, 104.35, 55.57, 50.73, 43.45.

Example 57: 6-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3- dihydroisoindol-1-one (1b-19)

¹H NMR (300 MHz, CDCI₃) δ 7.88 (d, J=1.5 Hz, 1 H), 7.81 (d, J=1.8 Hz, 1 H), 7.56 (dd, J=8.0, 1.7 Hz, 1 H), 7.45 (d, J=8.1 Hz, 1 H), 7.03-6.95 (m, 2H). 1³C NMR (75 MHz, CDCI₃) δ 165.94, 152.64, 139.74, 138.18, 135.16,

134.72, 134.19, 132.07, 124.10, 123.83, 117.97, 111.19, 104.34, 55.55, 50.83, 43.37.

Example 58: 2-(3-dimethylamino-4-methoxyphenyl)-2,3-dihydro- isoindol-1-one (1b-20)

¹H NMR (300 MHz, CDCI₃) δ 7.93 (d, J=7.3 Hz, 1 H), 7.67 (d, J=2.6 Hz, 1 H), 7.59 (d, J=6.8 Hz, 1 H), 7.58-7.49 (m, 2H), 7.22 (dd, J=8.7, 2.6 Hz, 1 H), 6.89 (d, J=8.7 Hz, 1 H), 4.85 (s, 2H), 3.92 (s, 3H), 2.86 (s, 3H).

¹³C NMR (75 MHz, CDCI₃) δ 167.51 , 149.62, 140.17, 134.15, 132.78, 131.89, 128.37, 124.00, 122.56, 114.30, 111.70, 111.27. Example 59: 2-(3-dimethylaminophenyl)-2,3-dihydroisoindol-1-one (1 b-21 )

¹H NMR (400 MHz, CDCI₃) δ 7.97 (d, J=7.6 Hz, 1 H), 7.72 (td, J=7.5, 1.0 Hz, 1 H), 7.59 (t, J=7Λ Hz, 1 H), 7.50 (dd, J=7.0, 0.7 Hz, 1 H), 6.68 (d, J=8.3 Hz, 1 H), 6.57 (s, 1 H), 6.10 (d, J=2.4 Hz, 1 H), 6.07 (dd, J=8.5, 2.5 Hz, 1 H), 3.93 (s, 2H), 2.93 (s, 6H).

¹³C NMR (100 MHz, CDCI₃) δ 170.44, 152.41 , 148.01 , 147.48, 133.78, 129.21 , 129.03, 127.16, 125.79, 123.86, 108.95, 103.32, 100.12, 80.79, 51.30, 40.29.

Example 60: 5-methoxy-2-(4-methylaminophenyl)-2,3-dihydro- isoindol-1-one (1b-23)

¹H NMR (300 MHz, DMSO-c/₆) δ 2.67 (s, 3H), 3.85 (s, 3H), 4.82 (s, 2H), 5.59 (s, NH), 6.57 (d, J=8.3 Hz, 2H), 7.05 (d, J=7.6 Hz, 1 H), 7.16 (s, 1 H), 7.51 (d, J=8.4 Hz, 2H)₁ 7.62 (d, J=7.5 Hz, 1H).

Example 61 : 5-(2-fluoroethoxy)-2-(4-methylaminophenyl)-2,3-dihydro- isoindol-1-one (1b-27) 1H NMR (300 MHz, DMSO-d₆) δ 2.68 (d, J=4.7 Hz, 3H), 4.30 (m, 1 H),

4.40 (m, 1 H), 4.70 (m, 1 H), 4.83 (m, 3H), 5.59 (m, NH), 6.58 (d, J=8.2 Hz, 2H), 7.09 (d, J=8.4 Hz, 1 H), 7.20 (s, 1 H), 7.52 (d, J=8.0 Hz, 2H), 7.64 (d, J=8.6 Hz, 1 H).

Example 62: 5-(2-fluoroethoxy)-2-(4-dimethylaminophenyl)-2,3- dihydroisoindol-1-one (1b-28)

¹H NMR (300 MHz, DMSOd₆) δ 2.88 (s, 6H), 4.29 (m, 1 H), 4.39 (m, 1 H), 4.69 (m, 1H), 4.86 (m, 3H), 6.78 (d, J=8.9 Hz, 2H), 7.09 (dd, J=1.9, 8.5 Hz, 1 H), 7.20 (d, J=1.9 Hz, 1 H), 7.62 (d, J=8.7 Hz, 2H), 7.68 (d, J=8.9 Hz, 1 H).

Example 63: 5-(3-fluoropropoxy)-2-(4-aminophenyl)-2,3-dihydro- isoindol-1-one (1 b-29)

¹H NMR (300 MHz, DMSO-d₆) δ 2.10 (t, J=5.6, 6.6 Hz, 1 H), 2.19 (t, J=5.5, 6.0 Hz, 1 H), 4.17 (t, J=6.0, 6.5 Hz, 2H), 4.55 (t, J=5.3, 5.9 Hz, 1 H),

4.71 (t, J=5.3 Hz, 1 H), 4.81 (s, 2H), 5.03 (s, NH₂), 6.60 (d, J=8.6 Hz, 2H),

7.06 (d, J=8.4 Hz, 1 H), 7.18 (s, 1 H), 7.43 (d, J=8.6 Hz, 2H), 7.62 (d, J=8.6 Hz,

1H).

Example 64: 5-(3-fluoropropoxy)-2-(4-methylaminophenyl)-2,3-dihydro- isoindol-1-one (1b-30)

¹H NMR (300 MHz, DMSOd₆) δ 2.10 (t, J=5.9 Hz, 1H), 2.19 (t, J=6.0 Hz, 1 H), 2.68 (s, 3H), 4.18 (t, J=6.0 Hz, 2H), 4.55 (t, J=5.7 Hz, 1 H), 4.71 (t, J=5.7 Hz, 1 H), 4.82 (s, 2H), 5.59 (m, NH), 6.58 (d, J=8.5 Hz, 2H), 7.07 (d, J=8.0 Hz, 1 H), 7.18 (s, 1H), 7.52 (d, J=8.0 Hz, 2H), 7.62 (d, J=8.0 Hz, 1 H).

Example 65: 5-(3-fluoropropoxy)-2-(4-dimethylaminophenyl)-2,3- dihydroisoindol-1-one (1b-31 )

¹H NMR (300 MHz, DMSO-d₆) δ 2.09 (t, J=6.1 Hz, 1 H), 2.18 (t, J=6.0 Hz, 1 H), 2.88 (s, 6H), 4.17 (t, J=6.2 Hz, 2H), 4.55 (t, J=6.1 Hz, 1 H), 4.70 (t, J=6.2 Hz, 1 H), 4.85 (s, 2H), 6.78 (d, J=9.1 Hz, 2H), 7.07 (dd, J=1.9, 8.4 Hz, 1 H), 7.19 (d, J=1.9 Hz, 1 H), 7.61 (d, J=9.0 Hz, 2H), 7.63 (d, J=8.3 Hz, 1 H).

Example 66: Pharmacological Activity Test - In Vitro Analysis of Binding Affinity to beta-Amyloid Fibrils, IC₅0 and Ki Values Using Gamma Rays of ¹²⁵I-TZDM

1 mg of beta-amyloid peptide (Aβi_-42 peptide, Bachem) was dissolved in 1 ml_ of dimethylsulfoxide (DMSO), to which 9 mL of phosphate buffer (pH

7.4) was added. The obtained solution was cultivated at 37⁰C for 60 minutes to obtain beta-amyloid fibril (Aβ-ι_-42 fibril), which was divided into 500 μL to each e-tube and kept in a freezer at -8O⁰C.

In order to check whether the beta-amyloid fibrils were formed well,

150 μL of thioflavine-T (Th-T) (5 μM) was added to 50 μL of the cultivated beta-amyloid peptide, and fluorescence density of the Th-T bound to the beta-amyloid fibrils were observed with a multi-label fluorescence counter

(LS-55 Luminescence spectrometer: Perkin Elmer) at λex/λem (450/480 nm).

As can be seen from Fig. 1, the fluorescent density of the group (control group) in which monomer beta-amyloid protein (Aβi_-4o) was bound with Th-T was 2,134, whereas that of the group in which beta-amyloid fibril (Aβ-ι_-42 fibril) was bound with Th-T was 176,619±22, 605, which is quite high. Thus, it was confirmed that beta-amyloid fibrils were formed.

In order to obtain a dissociation constant (K_d) of 2-(4'- dimethylaminophenyl)-6-[¹²⁵l]iodobenzothiazole (¹²⁵I-TZDM), beta-amyloid fibril (Aβ-ι_-42 fibril) at a concentration 10 nM (final reaction concentration) was prepared in a borosilicate glass tube of 12 mm x 75 mm, to which 50 μL (0.046-5.9 pM) of TZDM labeled with ¹²⁵I was added, and the resultant was then cultivated at room temperature for three hours after adjusting its final volume to 1 ml. with 10% ethanol. After the cultivation of three hours, ¹²⁵I-TZDM which was bound with the beta-amyloid fibrils and ¹²⁵I-TZDM which was not bound with the beta-amyloid fibrils were separated using a cell harvester (Brandel M-24R). Nonspecific binding was performed with 2 μM of Th-T, counting was performed with a gamma counter (Cobra-2), and then a dissociation coefficients (K_d) were determined by GraphPad Prism (GraphPad Software, San Diego, CA). As shown in Fig. 2, the dissociation coefficients (K_d) obtained with beta-amyloid fibrils and ¹²⁵I-TZDM was 0.13 nM.

In an experiment of binding affinity to beta-amyloid fibrils, 850 μL of 10% ethanol was put into a borosilicate glass tube of 12 mm x 75 mm, to which 50 μL of beta-amyloid fibril (Ap_1-42 fibril) was added (the final reaction concentration was 11.5 nM), and then 50 μL (the concentration of the final reaction solution was 1 nM) of the compound according to the Examples of the present invention was added thereto. 50 μL of ¹²⁵I-TZDM (the final reaction concentration was 0.05 nM) was added to the resultant, which was then cultivated for three hours. After the cultivation of three hours, ¹²⁵I-TZDM which was bound with beta-amyloid fibrils and ¹²⁵I-TZDM which was not bound with beta-amyloid fibrils were separated with a cell harvester (Brandel M-24R). Nonspecific binding was performed with 2 μM of Th-T and counting was performed using a gamma counter.

Binding affinity of the compounds according to examples of the present invention to beta-amyloid fibrils and IC₅₀ values with respect to inhibition of the association of beta-amyloid fibrils with ¹²⁵I-TZDM were determined, and inhibition coefficients (Kj) were determined according to Cheng-Prusoff formula (Ki = IC₅₀/(1+[L]/K_d) [See Cheng, Y.; Prusoff, W. H. Biochem. Pharmacol, 22, 3099(1973)] using GraphPad Prism (GraphPad Software, San Diego, CA). N-Methyl-[¹¹C]2-(4'-methylaminophenyl)-6-hydroxybenzothiazole (Pittsburgh Compound-B, PIB) [See Klunk, W. E. et al. Ann. Neurol. 55, 306(2004)] prepared by the inventors of the present invention was used as a control substance. Below Table 1 to Table 3 shows binding affinities and Kj values of the compounds of the present invention to beta-amyloid fibrils determined according to the methods as described above.

Table 1 :

a: Since binding affinity at 10 nM was low, test at 1nM was not performed.

Table 2:

a: Since binding affinity at 10 nM was low, test at 1 nM was not performed.

Table 3:

As can be seen from Tables 1 and 2, at the final concentration of 1 nM, three of the compounds according to the examples of the present invention exhibit superior binding affinities to beta-amyloid fibrils to the commercially available PIB (control substance) exhibiting strong binding affinity to beta-amyloid fibrils, and other three compounds exhibit almost the same binding affinities to beta-amyloid fibrils as that of PIB.

Meanwhile, as can be seen from Table 3, Ki values of nine compounds according to the present invention are superior to that of PIB (Ki=O.77 nM) as a control substance.

According to the present invention, the compounds of Formula 1 having excellent binding affinity to beta-amyloid fibrils and excellent efficiency of inhibiting the association between beta-amyloid fibrils and ¹²⁵I-TZDM, and its preparation method were provided. Because the compounds of the present invention have excellent binding affinities to beta-amyloid fibrils and the efficiency of inhibiting the association between beta-amyloid fibrils and ¹²⁵I-TZDM, the compounds according to the present invention can be favorably used for an early diagnosing, preventing or treating diseases related to beta-amyloid fibrils, including dementia.

Claims

1. A compound represented by the following Formula 1 or a pharmaceutically acceptable salt thereof: Formula 1

wherein A is C=O or CH₂ group; Ri is H, OH, a halogen atom selected from fluorine, chlorine, bromine and iodine, a CrC₈ alkyl group, a Ci-C₈ alkoxy group, a tosyloxy-CrC₈ alkoxy group, a mesyloxy-Ci-C₈ alkoxy group, a nosyloxy-CrC₈ alkoxy group, a CrC₈ alkoxy group substituted with halogen atoms selected from fluorine, chlorine, bromine and iodine, or a CrC₈ alkylamino group; R₂ is H, a halogen atom selected from fluorine, chlorine, bromine and iodine, or a CrC₈ alkyl group; and R₃ and R₄ are independently H, a CrC₈ alkoxy group or a CrC₈ alkylamino group.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1 , having one or more radioactive isotopes within its molecular structure.

3. The compound or a pharmaceutically acceptable salt thereof according to claim 1 , wherein the radioactive isotope is ¹¹C or ¹⁸F.

4. The compound or a pharmaceutically acceptable salt thereof according to claim 1 , which is selected from the group consisting of the following compounds:

5-dimethylamino-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-methoxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-hydroxy-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; 5-methyl-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-fluoro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-chloro-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-bromo-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; 5-dimethylamino-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3- dione;

5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione;

5-methyl-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione;

2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione; 5-fluoro-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione;

5-chloro-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione;

5-bromo-2-(4-dimethylamino-3-methoxyphenyl)isoindol-1 ,3-dione;

2-(3-dimethylamino-4-methoxyphenyl)isoindol-1 ,3-dione;

2-(3-dimethylaminophenyl)isoindol-1 ,3-dione; 5-methoxy-2-(4-aminophenyl)isoindol-1 ,3-dione;

5-methoxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione;

5-hydroxy-2-(4-aminophenyl)isoindol-1 ,3-dione;

5-hydroxy-2-(4-methylaminophenyl)isoindol-1 ,3-dione;

5-(2-fluoroethoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione; 5-(2-fluoroethoxy)-2-(4-methylaminophenyl)isoindol-1 ,3-dione;

5-(2-fluoroethoxy)-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione; 5-(3-fluoropropoxy)-2-(4-aminophenyl)isoindol-1 ,3-dione;

5-(3-fluoropropoxy)-2-(4-methylaminophenyl)isoindol-1 ,3-dione;

5-(3-fluoropropoxy)-2-(4-dimethylaminophenyl)isoindol-1 ,3-dione;

5-methoxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; 5-hydroxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

5-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

2-(4-dimethyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

5-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

5-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one; 5-bromo-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1 -one;

6-methyl-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

6-fluoro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

6-chloro-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

5-methoxy-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol- 1-one;

5-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1- one;

2-4(-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1-one;

5-fluoro-2-(4-dimethylamino-3-mθthoxyphenyl)-2,3-dihydroisoindol-1- one;

5-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1- one;

5-bromo-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1- one; 6-methyl-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1 - one; 6-fluoro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1- one;

6-chloro-2-(4-dimethylamino-3-methoxyphenyl)-2,3-dihydroisoindol-1- one; 2-(3-dimethylamino-4-methoxyphenyl)-2,3-dihydroisoindol-1-one;

2-(3-dimethylaminophenyl)-2,3-dihydroisoindol-1-one;

5-methoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one;

5-methoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1-one;

5-methoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one; 5-methoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1 -one;

5-(2-fluoroethoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1-one;

5-(2-fluoroethoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1-one;

5-(2-fluoroethoxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1-one

5-(3-fluoropropoxy-2-(4-aminophenyl)-2,3-dihydroisoindol-1 -one;

5-(3-fluoropropoxy-2-(4-methylaminophenyl)-2,3-dihydroisoindol-1-one; 5-(3-fluoropropoxy-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1- one;

5-methoxy-2-(4-¹¹methylaminophenyl)-2,3-dihydroisoindol-1-one; and 5-(2-¹⁸fluoroethoxy)-2-(4-dimethylaminophenyl)-2,3-dihydroisoindol-1 - one.

5. The compound or a pharmaceutically acceptable salt thereof according to claim 1 , which is a salt of an inorganic or organic acid selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, benzenesulfonic acid, and toluenesulfonic acid.

6. The compound or a pharmaceutically acceptable salt thereof according to claim 1 , which is used for the diagnosis, prevention or treatment of a degenerative cerebral disease.

7. A pharmaceutical composition for the diagnosis, prevention or treatment of a degenerative cerebral disease, comprising the compound or a pharmaceutically acceptable salt thereof according to claim 1.

8. The pharmaceutical composition according to claim 7, wherein the degenerative cerebral disease is Alzheimer's disease, cerebrovascular dementia or Parkinson's disease.

9. A preparation method of a compound represented by Formula 1 according to claim 1 , comprising reacting a compound represented by Formula 2a or 2b with an aromatic amine compound represented by Formula 3: Formula 1 :

Formula 2a:

Formula 2b:

Formula 3:

wherein A is C=O or CH₂ group; Ri is H, OH, a halogen atom selected from fluorine, chlorine, bromine and iodine, CrC₈ alkyl group, a CrCs alkoxy group, a tosyloxy-CrC₈ alkoxy group, a mesyloxy-Ci-C₈ alkoxy group, a nosyloxy-C-i-Cβ alkoxy group, a CrC₈ alkoxy group substituted with halogen atoms selected from fluorine, chlorine, bromine and iodine, or a Ci-C₈ alkylamino group; R₂ is H, a halogen atom selected from fluorine, chlorine, bromine and iodine, or a CrC₈ alkyl group; and R₃ and R₄ are independently H, a Ci-C₈ alkoxy group or a CrC₈ alkylamino group.

10. The preparation method according to claim 9, wherein the reaction is conducted in an acidic solvent.

11. The preparation method according to claim 10, wherein the acidic solvent is acetic acid, hydrochloric acid or p-toluenesulfonic acid.

12. The preparation method according to claim 9, further comprising a reduction of a product of the reaction of the compound of Formula 2a or 2b with the compound of Formula 3 using tin, zinc, iron, hydrazine, copper sulfate or tin chloride under an acid condition.

13. The preparation method according to claim 12, further comprising a reductive amination of a product of the reduction.

14. The preparation method according to claim 13, wherein the reductive amination is carried out with sodium methoxide, p-formaldehyde and sodium borohydride.

15. The preparation method according to claim 9, further comprising dealkylation of a product obtained from the reaction of the compound of Formula 2a or 2b with the compound of Formula 3 using boron tribromide.

16. The preparation method according to claim 15, further comprising a nucleophilic substitution of a product of the dealkylation with a compound having a general formula of X(CH₂)_nOR₆ (wherein RQ is a tosyl, mesyl, or nosyl group, n is 2 or 3 and X is a halogen atom selected from fluorine, chlorine, bromine and iodine).

17. The preparation method according to claim 12, further comprising a nucleophilic substitution of a product of the reduction with [¹¹C] methoxytriflate.

18. The preparation method according to claim 15, further comprising a nucleophilic substitution of a product of the dealkylation with a compound having a general formula of R₆O(CH₂)_POR₆ (wherein R₆ is a tosyl, mesyl, or nosyl group, and n is 2 or 3).

19. The preparation method according to claim 18, further comprising a nucleophilic substitution of a product of the nucleophilic substitution with tetrabutylammonium ¹⁸fluoride.