JP7008299B2 - Imaging and in vitro diagnostics for intractable neurological diseases - Google Patents

Description

The present invention relates to diagnostic imaging agents and in vitro diagnostic agents for tauopathy, particularly novel compounds and salts thereof useful for diagnosing Alzheimer's disease.

Alzheimer's disease is a disease characterized by progressive dementia that occurs from presenile to old age, and it is said that the number of patients in Japan is currently more than 4.6 million. Furthermore, it is expected that the number will surely increase in the future as the population ages. Advances in research on Alzheimer's disease are energetically advancing the development of radical therapeutic agents. In order to treat Alzheimer's disease using these new drugs, a non-invasive method for early and accurate diagnosis of Alzheimer's disease is indispensable.

The clinical symptoms of Alzheimer's disease are memory impairment, higher brain dysfunction (aphasia, apraxia, agnosia, constructional apraxia) and the like. The symptoms are often seen in other dementia diseases, and it is extremely difficult to make a definitive diagnosis of Alzheimer's disease based on clinical symptoms alone.

On the other hand, characteristic histopathological findings of Alzheimer's disease include amyloid plaque and neurofibrillary tangles. The main constituent of the former is amyloid β protein with a β-sheet structure, and that of the latter is hyperphosphorylated tau protein. It is known that in Alzheimer's disease, the above-mentioned pathological tissue changes such as accumulation of aggregated amyloid β protein have started long before the onset of clinical symptoms.

From this point of view, in recent years, research on radiocontrast agents for positron emission tomography (PET) and single photon tomography (SPECT) that selectively bind to amyloid β protein in the brain has been advanced. However, since they use radionuclides such as ¹¹ C, ¹³ N, ¹⁵ O, and ¹⁸ F, there are concerns about side effects due to radiation damage, and it is necessary to install a cyclotron facility nearby, and the price of reagents is extremely high. Is a problem. Therefore, a diagnostic method that does not use radionuclides is desired.

Magnetic resonance imaging (MRI) is one of the diagnostic methods that does not use radionuclides. So far, the present inventors and a group of RIKEN have reported that they have succeeded in imaging geriatric spots using a fluorine nuclear magnetic resonance imaging method (fluorine MR imaging method) (Patent Documents 1 to 5). , Non-Patent Document 1).

Diseases with abnormal deposits of tau protein are called tauopathy. Although hyperphosphorylated tau protein accumulation is thought to occur later than amyloid β protein accumulation, neurofibrillary tangles are more closely associated with disease severity than amyloid β protein accumulation. It is believed to be related.

Therefore, research on radioactive contrast media for PET and SPECT that bind to tau protein is underway, and such contrast media are reported in, for example, Patent Documents 6 to 9. Although the radionuclides have the above-mentioned problems, there have been no reports of successful imaging of neurofibrillary tangles using MRI.

International Publication No. 2005/042461 International Publication No. 2007/111179 Japanese Unexamined Patent Publication No. 2009-67762 International Publication No. 2010/098502 International Publication No. 2014/109296 International Publication No. 2004/054978 International Publication No. 2005/016888 International Publication No. 2012/05731 International Publication No. 2014/097474

Higuchi M, Iwata N, Matsuba Y, Sato K, Sasamoto K, Saido T: 19F and 1H MRI detection of amylid β plaques in vivo, Nature Neuroscience, 8 (4), 527-533, 2005.

An object of the present invention is to provide a substance having both high binding specificity to tau protein and high detection sensitivity, which is suitable as a contrast agent for MRI diagnosis of tauopathy.

As a result of diligent research to achieve the above object, the present inventor has a high binding specificity for tau protein in a compound having a certain chemical structure, and by using the compound as a contrast agent, neurofibrillary tangles are changed by MRI. Was found to be able to be imaged.

That is, the present invention relates to a compound or a salt thereof selected from the aniline derivative represented by the formula (1) and the naphthalene derivative represented by the formula (2).

(In the formula, R ¹ and R ² independently represent a hydrogen atom or an alkyl, respectively.
One of A ¹ and A ² indicates a hydrogen atom, and the other is a fluorine atom-substituted or unsubstituted alkoxy or

Is shown. Here, R ³ indicates fluorine atom-substituted or unsubstituted ethylene, R ⁴ indicates fluorine atom-substituted or unsubstituted ethyl, and n indicates an integer of 1 to 10. If n represents an integer greater than or equal to 2, R ³ may be the same or different. )

Further, the present invention relates to a tauopathy diagnostic imaging agent and a PET diagnostic imaging agent containing a compound selected from the aniline derivative of the formula (1) and the naphthalene derivative represented by the formula (2) or a salt thereof as an active ingredient. , And in vitro diagnostic agents.

The alkyl group of R ¹ and R ² may be a linear or branched C _{1 to 6} alkyl, and specific examples thereof include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl. However, linear or branched C _1-3 alkyl is preferable.

The alkoxy groups of A ¹ and A ² may be linear or branched C _{1 to 10} alkoxy groups, and specific examples thereof include methoxy, ethoxy, propoxy, butoxy, pentyloxy, and hexyloxy. C _{1 to 6} alkoxy in the form of a straight chain or a branched chain is preferable.

Of A ¹ and A ² , preferably one is a hydrogen atom and the other is a hydrogen atom.

Is.

n is an integer of 1 to 10, preferably an integer of 5 to 8, more preferably 5 or 6, and even more preferably 6 in terms of detection accuracy.

The geometric isomers in the double bond of the aniline derivative of the formula (1) include both cis and trans isomers.

The aniline derivative of the formula (1) and the naphthalene derivative of the formula (2) are preferably labeled. Examples of the label include fluorescent substances, radionuclides and the like, and radionuclides are preferable. As the radionuclide, positron emitting nuclides such as ¹¹ C, ¹³ N, ¹⁵ O, ¹⁸ F, ⁶² Cu, ⁶⁸ Ga, and ⁷⁶ Br can be used as diagnostic imaging agents for PET. Of these, ¹⁸ F is particularly preferable. The labeling position of the aniline derivative of the formula (1) and the naphthalene derivative of the formula (2) in the radionuclide can be any position in the compound. It is also possible to replace the hydrogen in the compound with a positron emitting nuclide.

Generally, the nuclide is produced by a device called a cyclotron or a generator. Those skilled in the art can appropriately select a production method and an apparatus according to the nuclide produced. Methods for producing compounds labeled with these radiation nuclides are well known in the art. Typical methods include a chemical synthesis method, an isotope exchange method and a biosynthesis method.

The salt of the aniline derivative of the formula (1) and the naphthalene derivative of the formula (2) may be any pharmaceutically acceptable salt, for example, a salt salt, a sulfate, a nitrate, a mineral salt such as a phosphate; Organic acid salts such as tartrate, acetate, citrate, malate, fumarate, maleate, benzoate, glycolate, amber, p-toluenesulfonate, methanesulfonate, etc. And so on. In addition, some of these salts have water of crystallization.

The compound of the present invention has high affinity for tau protein and high blood-brain barrier permeability, and is useful as an active ingredient of a diagnostic imaging agent for tauopathy, for example, an active ingredient of an MRI contrast agent and a PET contrast agent. It is also useful as an active ingredient of an in vitro diagnostic agent for diagnosing tauopathy. Therefore, the compound of the present invention enables early diagnosis of tauopathy such as Alzheimer's disease.

3 is a fluorescence image of a postmortem brain section of a patient with Alzheimer's disease in Test Example 1. Scale bar: 100 μm It is a fluorescence and staining image of the brain section of the tau gene-modified mouse in Test Example 2. A, C, D, E: Fluorescent image, B: Phosphorylated tau antibody stained image, A, B: Compound 1, C: Compound 2, D: Compound 3, E: Compound 4 Head ¹ H MRI images (A, C) and fluorine MRI images (B, D) when compound 2 in Test Example 3 was administered to tau gene-modified mice and wild-type mice. Head ¹ H MRI images (A, C) and fluorine MRI images (B, D) when compound 3 in Test Example 4 was administered to tau gene-modified mice and wild-type mice. 3 is a fluorescence image of a brain section (A, C: hippocampus, B, D: cerebral cortex) when compound 2 or compound 3 in Test Example 5 was administered to a tau gene-modified mouse. A, B: Compound 2, C, D: Compound 3 3 is a fluorescence image of a postmortem brain section of a patient with Alzheimer's disease in Test Example 6. Scale bar: 50 μm 1H MRI image ( ^A ) and fluorine MRI image (B) of the head when compound 6 in Test Example 7 was administered to a tau gene-modified mouse. 3 is a fluorescent image and a stained image of a brain section when Compound 6 in Test Example 8 was administered to a tau genetically modified mouse. A: Fluorescent image, B: Phosphorylated tau antibody stained image Scale bar: 100 μm

Among the compounds of the formula (1) or salts thereof, a method for producing the compound of the formula (I) or a salt thereof is shown below.

The compound of the formula (I) or a salt thereof can be produced by the following method.

The compound of the formula (I) can be produced by reacting the compound of the formula (II) with a cinnamaldehyde derivative.

(In the equation, R ¹ , R ² and n are as described above.)

The solvent for this reaction is aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether, ligroin and petroleum benzine; diethyl ether, dipropyl ether and dibutyl ether. , Ethers such as tetrahydrofuran and dioxane; Acid amides such as dimethylformamide and dimethylacetamide; Sulfoxides such as dimethylsulfoxide; Sulfones such as sulfolane; Phosphoric acid amides such as hexamethylphosphorylamide; Examples thereof include alcohols such as butanol and mixed solvents thereof.

In order to promote this reaction, it is desirable to add a base, which is a hydroxide of an alkali metal such as sodium hydroxide and potassium hydroxide; and an alkaline earth metal such as calcium hydroxide and barium hydroxide. Hydroxide; Alkali metals such as lithium, sodium, potassium; Metal alkoxides such as sodium methoxyd, sodium ethoxydo, potassium tert-butoxide; Metal hydrides such as lithium hydride, sodium hydride, calcium hydride, etc. be able to. The base can be used in 1 to 5 times mol, preferably 1 to 2 times mol, with respect to compound (II).

This reaction can be carried out at 0 to 50 ° C., preferably 0 to 20 ° C., and the reaction time is usually about 0.5 to 6 hours.

The cinnamaldehyde derivative can be used in 1 to 5 times mol, preferably 2 to 3 times mol, with respect to the compound of formula (II).

The compound of the formula (II) can be produced by reacting the compound of the formula (IV) with the compound of the formula (III).

(In the formula, m is as described above)

The solvent for this reaction is aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether, ligroin and petroleum benzine: diethyl ether, dipropyl ether and dibutyl ether. , Ethers such as tetrahydrofuran and dioxane; nitriles such as acetonitrile and propionitrile; ketones such as acetone and methyl ethyl ketone; acid amides such as dimethylformamide and dimethylacetamide; And these mixed solvents can be mentioned.

In order to promote this reaction, it is desirable to add a base, which is a carbonate of a metal such as sodium carbonate and potassium carbonate; and water of an alkali metal such as lithium hydroxide, sodium hydroxide and potassium hydroxide. Oxides; Alkaline earth metal hydroxides such as calcium hydroxide and barium hydroxide; Metal hydrides such as sodium hydride and calcium hydride; Metal alkoxides such as sodium methoxydo, sodium ethoxydo and potassium tert-butoxide ; Metal hydrides such as lithium hydride, sodium hydride, calcium hydride and the like can be mentioned. The base can be used in 1 to 5 times mol, preferably 1 to 3 times mol, with respect to the compound of formula (III).

This reaction can be carried out at 0 to 150 ° C., preferably 20 to 100 ° C., and the reaction time is about 5 to 50 hours.

The compound of formula (III) can be used 1 to 5 times mol, preferably 2 to 3 times mol, of the compound of formula (IV).

The compound of formula (IV) can be produced by brominating the compound of formula (V).

(In the formula, m is as described above.)

Solvents for this reaction include saturated hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether, ligroin and petroleum benzine; diethyl ether, dipropyl ether and dibutyl ether, Ethers such as tetotahydrofuran and dioxane; acid amides such as dimethylformamide and dimethylacetamide; sulfoxides such as dimethylsulfoxide; phosphate amides such as hexamethylphosphorylamide; Can be mentioned.

This reaction is usually preferably carried out at 50 to 150 ° C., and ammonium bromide can be used in 2 to 5 times mol, preferably 2 to 3 times mol, with respect to the compound of formula (V).

The compound of formula (V) can be produced by p-toluenesulfonylation of the compound of formula (VI).

(In the formula, m is as described above.)

This reaction involves a two-step reaction of elimination of protecting groups by acid hydrolysis and tosylation of the produced alcohol.

First, regarding acid hydrolysis, in addition to water as a solvent, alcohols such as methanol, ethanol, propanol and butanol; ethers of hydrated acetonitrile and hydrated dioxane; acid amides such as hydrated dimethylformamide and hydrated dimethylacetamide; Nitriles such as hydrous acetonitrile and hydropropionitrile; sulfoxides such as hydrous dimethyl sulfoxide and mixed solvents thereof can be mentioned.

It is desirable to add an acid to promote this reaction, and examples of the acid include mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, and perchloric acid. The acid used should be about 0.01 to 1 times the molar amount of the compound of the formula (VI).

This reaction can be carried out at 0 to 100 ° C., preferably 0 to 50 ° C., and the reaction time is usually about 0.5 to 50 hours.

Next, regarding tosylation of alcohol, this reaction is carried out by reacting the alcohol obtained by hydrolysis with p-toluenesulfonyl chloride.

The solvent for this reaction is aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether, ligroin and petroleum benzine; diethyl ether, dipropyl ether and dibutyl ether. , Hydrocarbons, dioxane and other ethers; acetonitrile, propionitrile and other nitriles; dimethylformamide, dimethylacetamide and other acid amides; dimethylsulfoxide and other sulfoxides; chloroform, dichloromethane, carbon tetrachloride, 1,2-dichloroethane Such as halogenated hydrocarbons and mixed solvents thereof can be mentioned.

It is desirable to add a base to promote this reaction, and the bases include organic bases such as triethylamine, pyridine, N-methylmorpholin, N, N, -dimethylaniline, and diisopropylethylamine; sodium carbonate, potassium carbonate, etc. Alkaline metal carbonates; Examples thereof include alkali metal bicarbonates such as sodium hydrogencarbonate and potassium hydrogencarbonate. The base can be used in 1 to 5 times mol, preferably 1 to 2 times mol, with respect to p-toluenesulfonyl chloride.

This reaction can be carried out at 0 to 100 ° C., preferably 0 to 50 ° C., and the reaction time is about 1 to 50 hours.

The p-toluenesulfonyl chloride can be used in 1 to 5 times mol, preferably 2 to 3 times mol, with respect to the compound of formula (VI).

The compound of the formula (VI) can be produced by reacting the compound of the formula (VIII) with the compound of the formula (VII). The compound of the formula (VIII) can be produced by a known method.

(In the formula, m is as described above.)

The solvent for this reaction is aromatic hydrocarbons such as benzene, toluene and xylene; aliphatic hydrocarbons such as pentane, hexane, heptane, petroleum ether, ligroin and petroleum benzine; diethyl ether, dipropyl ether and dibutyl ether. , Ethers such as tetrahydrofuran and dioxane; Acid amides such as dimethylformamide and dimethylacetamide; Sulfoxides such as dimethylsulfoxide; Sulfones such as sulfolane; Phosphoric acid amides such as hexamethylphosphorylamide and mixed solvents thereof. be able to.

It is desirable to add a base to promote this reaction, and the bases include alkali metals such as lithium, sodium and potassium; metal hydrides such as lithium hydride, sodium hydride and calcium hydride; potassium tert- Examples thereof include metal alkoxides such as butoxide. The base can be used in 1 to 5 times mol, preferably 1 to 3 times mol, with respect to the compound of formula (VII).

This reaction can be carried out at 0 to 150 ° C., preferably 0 to 100 ° C., and the reaction time is 1 to 50 hours.

The compound of formula (VII) can be used in 1 to 10 times mol, preferably 1 to 3 times mol, with respect to the compound of formula (VIII).

The compound of formula (IX) or a salt thereof can be produced by using 5-hydroxy-2-methylbenzoxazole instead of 6-hydroxy-2-methylbenzoxazole in the above method.

(In the formula, R ¹ , R ² and m are as described above.)

Among the compounds of formula (1) or salts thereof, the compound in which A ¹ or A ² is alkoxy or a salt thereof is obtained by using an alkyl halide instead of the compound of formula (IV) in the above method. Can be manufactured.

The compound of the formula (2) or a salt thereof can be produced according to the method described in Examples described later or by appropriately modifying the above method with reference to the above method.

The compounds of formulas (1) and (2) obtained by the above-mentioned production method and the method incidental thereto can be obtained by known means, for example, concentration, concentration under reduced pressure, distillation, fractional distillation, transsolution, solvent extraction, crystallization, recrystallization. It can be isolated and purified by crystallization, chromatography and the like.

When the compounds of the formulas (1) and (2) are obtained in free form, salts can be formed by a usual method.

Table 1 shows some examples of the compounds of formulas (1) and (2).

Table 1

The compound of the compound of the formula (1) and the formula (2) or a salt thereof can be used as a diagnostic imaging agent for MRI of tauopathy, an diagnostic imaging agent for PET, and an in vitro diagnostic agent.

Desirable embodiments of the present invention are:
<1> An MRI diagnostic imaging agent for tauopathy containing a compound of formula (1) or formula (2) or a salt thereof as an active ingredient.
<2> A diagnostic imaging agent for PET of tauopathy containing the compound of the formula (1) or the formula (2) or a salt thereof as an active ingredient.
<3> An in vitro diagnostic agent for diagnosing tauopathy containing the compound of the formula (1) or the formula (2) or a salt thereof as an active ingredient.
<4> The diagnostic imaging according to any one of <1> to <3>, wherein the tauopathy is Alzheimer's disease, progressive supranuclear palsy, corticobasal degeneration, frontotemporal dementia or Pick disease. Drugs or in vitro diagnostics.
<5> A method for diagnosing tauopathy, which comprises the step of administering the diagnostic imaging agent according to <1> or <2>.
<6> A method for diagnosing tauopathy, which comprises a step of detecting tau protein in a sample using the in vitro diagnostic agent according to <3>.

As a compound to be administered to a living body, it is desirable that the compound has high water solubility, and among the compounds of the formulas (1) and (2), a compound having a salt is more preferable.

When the compound of formula (1) or formula (2) or a salt thereof is used as a diagnostic imaging agent, the compound of the present invention can specifically detect neurofibrillary tangles in the brain. In particular, when tau protein is detected non-invasively using ¹⁹ F-MRI, its detection sensitivity depends on the number of fluorine atoms, and it is desirable that the number of F atoms is large. Therefore, it is desirable that the compound of the formula (1) or the formula (2) or a salt thereof used as an diagnostic imaging agent for MRI is substituted with one or more, preferably 1 to 3 fluorine atoms. Further, the compound of the formula (1) or the formula (2) or a salt thereof used as a diagnostic imaging agent for PET is usually labeled with a positron emitting nuclide.

When the compound of the formula (1) or the formula (2) or a salt thereof is used as a diagnostic imaging agent, the administration thereof may be local or systemic. The administration method is not particularly limited, and it is administered orally or parenterally. Parenteral routes of administration include subcutaneous, intraperitoneal, intravenous, arterial or spinal fluid injections, infusions and the like.

The diagnostic imaging agent containing the compound of the formula (1) or the formula (2) or a salt thereof contains a pharmaceutically acceptable form suitable for administration to a human and a physiologically acceptable additive. Such compositions may optionally be pharmaceutically acceptable diluents, buffers, solubilizers (eg, cyclodextrin, polyethylene glycol, Pluronic ™, Tween ™, Cremofol ™ or phospholipids). Surfactants), soothing agents and the like can be added and, if necessary, contain components such as pharmaceutically acceptable solvents, stabilizers or antioxidants (eg, ascorbic acid). obtain. The dose of the compound of the present invention is appropriately selected depending on the usage, the age of the patient, the sex and other conditions, and the degree of the disease.

When the compound of the formula (1) or the formula (2) or a salt thereof is used as an in vitro diagnostic agent, the tau protein in the sample can be specifically detected by the compound of the present invention. The tau protein in the sample can be measured by measuring the fluorescence of the compound of the present invention bound to the tau protein contained in the sample. Examples of in vitro diagnostic agents include blood, cerebrospinal fluid, and body fluids such as tears, saliva, nasal discharge, and urine. The in vitro diagnostic agent may contain other additives to the extent that it does not interfere with the function of the in vitro diagnostic agent. The amount of the compound of the present invention to be used may be any amount as long as the binding amount is sufficient to the extent that the presence of tau protein can be determined, and is appropriately selected depending on the conditions such as the type and concentration of the sample.

Examples of tauopathy include, in addition to Alzheimer's disease, progressive supranuclear palsy (PSP), corticobasal degeneration, frontotemporal dementia, Pick disease and the like.

Next, synthetic examples and test examples according to the present invention will be described, but the present invention is not limited thereto.

[Synthesis Example 1] Synthesis of 1- (6-methoxybenzoxazole-2-yl) -4- (4'-dimethylaminophenyl) pig-1,3-diene (Compound 1)

(1) Add 1.24 mL (20 mmol) of methyl iodide to a solution of 1.49 g (10 mmol) of 6-hydroxy-2-methylbenzoxazole in acetone (40 mL), and further add 4.14 g (30 mmol) of potassium carbonate. The mixture was vigorously stirred at room temperature for 6 hours. After completion of the reaction, the insoluble solid matter in the reaction solution was filtered, and the filtrate was concentrated to dryness under reduced pressure. The obtained residue was extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 1: 3) and 6-methoxy-2-methylbenzoxazole 1.585 g (97%). Was obtained as colorless crystals (mp 54 ° C.).
¹ HNmr (CDCl ₃ ) δ2.60 (3H, s), δ3.85 (3H, s) δ6.90 (1H, dd, J = 2.4Hz, 8.6Hz), δ7.01 (1H, d, J = 2.4Hz), δ7.51 (1H, d, J = 8.6Hz)

(2) 163 mg (1 mmol) of 6-methoxy-2-methylbenzoxazole obtained in the above step (1) and 201 mg (1 mmol) of 4-dimethylaminocinnamaldehyde were dissolved in DMF (1.5 mL). The solution was ice-cooled and 168 mg (1.5 mmol) of potassium t-butoxide was added in small portions with stirring. After the addition was completed, the reaction solution was stirred at room temperature for 1.5 hours, and then the reaction solution was diluted with ethyl acetate. The ethyl acetate solution was washed with water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: dichloromethane), recrystallized from ethyl acetate, and (E, E) -1- (6-methoxybenzo). Oxazole-2-yl) -4- (4'-dimethylaminophenyl) porcine-1,3-diene 157 mg (45.5%) was obtained as dark reddish brown crystals (mp160-161 ° C.).
¹ HNmr (CDCl ₃ ) δ3.01 (6H, s), δ3.87 (3H, s), δ6.47 (1H, d, J = 15.5Hz), δ6.69 (2H, d, J = 8.9Hz) ), δ6.8-6.85 (2H), δ6.90 (1H, dd, J = 8.6Hz, 2.4Hz), 7.03 (1H, d, J = 2.4Hz), δ7.39 (2H, d, J = 8.9Hz), δ7.4-7.5 (1H), δ7.53 (1H, d, J = 8.6Hz)

[Synthesis Example 2] 1- [6- (3', 6', 9', 12', 15'-pentaoxa-17', 17', 17'-trifluoroheptadecanyloxy) benzoxazole-2- Il] -4- (4'-dimethylaminophenyl) porcine-1,3-diene (Compound 2) synthesis

(1) Weigh 0.49 g (12.2 mmol) of 60% sodium hydride in a flask in a nitrogen stream, and while cooling with ice, 1.92 g (13.3) of 2- (2', 2', 2'-trifluoroethoxy) ethanol. A solution of mmol) in THF (12 mL) was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 1 hour. The reaction was ice-cooled again and 1- (2H-tetrahydropyran-2'-yloxy) -11- (4'-p-toluenesulfonyloxy) -3,6,9-trioxaundecane 4.43 g (10.2 mmol). The THF (8 mL) solution was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 20 hours. The reaction mixture was diluted with ethyl acetate and washed with a small amount of water and saturated brine. After drying with anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 200) and 1- (2H-tetrahydropyran-). 2'-Iloxy) -17,17,17-trifluoro-3,6,9,12,15-pentaoxaheptadecane 4.07 g (98%) was obtained as an oil.

(2) 1- (2H-tetrahydropyran-2'-yloxy) -17,17,17-trifluoro-3,6,9,12,15-pentaoxaheptadecane 4.07 obtained in the above step (1). 1M-hydrochloric acid (0.1 mL) was added to a solution of g (10 mmol) in ethanol (40 mL), and the mixture was allowed to stand at room temperature for 13 hours. Saturated aqueous sodium hydrogen carbonate solution was added for neutralization, and the precipitated precipitate was filtered off. The filtrate was concentrated to dryness under reduced pressure, and the obtained residue was extracted with dichloromethane. After drying the extract with anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, the obtained residue (3.21 g) was dissolved in dichloromethane (25 mL), and 2.5 g (13 mmol) of p-toluenesulfonyl chloride was added. After that, 2.1 mL (15 mmol) of triethylamine was added little by little while cooling with ice. The reaction mixture was stirred at room temperature for 22 hours, diluted with dichloromethane, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1- (4'-p toluenesulfonyloxy) -17,17, 17-Trifluoro-3,6,9,12,15-pentaoxaheptadecane 4.02 g (85%) was obtained as a colorless oil.

(3) 1- (4'-p-toluenesulfonyloxy) -17,17,17-trifluoro-3,6,9,12,15-pentaoxaheptadecane 4.02 g obtained in the above step (2). A mixture of 1.67 g (17 mmol) ammonium bromide in a solution of (8.5 mmol) in DMF (15 mL) was heated to 80-85 ° C. for 6 hours with vigorous stirring. The solvent was distilled off under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 200) and 1-bromo-17,17,17-trifluoro-3,6. , 9,12,15-Pentaoxaheptadecane 3.04 g (94%) was obtained as a colorless oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.47 (2H, t, J = 6Hz), δ3.6-3.9 (18H), δ3.91 (2H) , q, J = 9Hz)

(4) 1-bromo-17,17,17-trifluoro-3,6,9,12,15-pentaoxaheptadecane 0.80 g (2.1 mmol) and 6-hydroxy- obtained in the above step (3). To a solution of 0.44 g (2.9 mmol) of 2-methylbenzoxazole (III) in acetone (8 mL), add 1.2 g (8.7 mmol) of potassium carbonate and 33 mg (0.2 mmol) of potassium iodide. The added mixture was refluxed for 10 hours with vigorous stirring. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 2: 1) and 2-methyl-6- (3', 6', 9'). , 12', 15'-pentaoxa-17', 17', 17'-trifluoroheptadecanyloxy) benzoxazole 0.90 g (95%) was obtained as an oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ2.60 (3H, s), δ3.6-3.8 (16H), δ3.88 (2H, m), δ3.90 (2H, q, J = 9Hz), 4.15 (2H, m), δ6.92 (1H, dd, J = 2.1Hz, 8.6Hz), δ7.03 (1H, d, J = 2.1Hz) , δ7.50 (1H, d, J = 8.6Hz)

(5) 2-Methyl-6- (3', 6', 9', 12', 15'-pentaoxa-17', 17', 17'-trifluoroheptadecanyl obtained in the above step (4) Oxy) In a solution of 0.27 g (0.6 mmol) of benzoxazole and 0.21 g (1.2 mmol) of 4-dimethylaminocinnamaldehyde in DMF (2.5 mL), potassium t-butoxide 0.20 g (1.8 mmol) while ice-cooling. Was added little by little. After the addition was completed, the reaction solution was stirred at room temperature for 4 hours. The reaction mixture was diluted with ethyl acetate, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 2: 1) and 1- [6- (3', 6', 9', 12', 15'-Pentaoxa-17', 17', 17'-Trifluoroheptadecanyloxy) Benzoxazole-2-yl] -4- (4'-Dimethylaminophenyl) Buta-1,3-diene 0.22 g (60%) was obtained as a dark red oil (mixture of geometric isomers).
¹⁹ FNmr (CDCl ₃ ) δ-75.55 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.01, 3.02, 3.03 (singlet, 6H in total), δ3.6-3.8 (16H), δ3. 8-4.0 (4H), 4.25 (2H, m), δ6.4-7.1 (7H), δ7.3-7.6 (4H)

[Synthesis Example 3] 1- [6- (3', 6', 9', 12', 15', 18'-hexaoxa-20', 20', 20'-trifluoroicosanyloxy) benzoxazole-2 -Il] -4- (4'-dimethylaminophenyl) porcine-1,3-diene (Compound 3) synthesis

(1) Weigh 0.40 g (10 mmol) of 60% sodium hydride in a flask in a nitrogen stream, and while cooling with ice, 1.44 g (10) of 2- (2', 2', 2'-trifluoroethoxy) ethanol. A solution of mmol) in THF (12 mL) was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 1 hour. The reaction was ice-cooled again and 1- (2H-tetrahydropyran-2'-yloxy) -14- (4'-p-toluenesulfonyloxy) -3,6,9,12-tetraoxatetradecane 4.29 g (9). A solution of mmol) in THF (8 mL) was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 15 hours. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was extracted with ethyl acetate. The extract was washed with a small amount of water and then with a small amount of saturated brine, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure using a rotary evaporator, and the obtained residue was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1:50) and 1- (2H-tetrahydropyran-2'- Iloxy) -20,20,20-trifluoro-3,6,9,12,15,18-hexaoxaicosan 3.64 g (90%) was obtained as a colorless oil.

(2) 1- (2H-tetrahydropyran-2'-yloxy) -20,20,20-trifluoro-3,6.9,12,15,18-hexaoxaicosan 3.64 obtained in the above step (1). Add 1M-hydrochloric acid (0.1 mL) to a solution of g (8.12 mmol) in ethanol (25 mL), leave it at room temperature for 24 hours, then add saturated aqueous sodium hydrogen carbonate solution to neutralize and precipitate. Was filtered out. The filtrate was concentrated to dryness under reduced pressure using a rotary evaporator, and the obtained residue was extracted with dichloromethane. After drying the extract with magnesium sulfate, the residue (2.76 g) obtained by concentrating and drying under reduced pressure was dissolved in dichloromethane (20 mL), and 1.9 g (9.9 mol) of p-toluenesulfonyl chloride was added. , 1.5 mL (10.8 mmol) of triethylamine was added little by little while cooling with ice. The reaction mixture was stirred at room temperature for 24 hours, diluted with dichloromethane, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1- (4'-p-toluenesulfonyloxy) -20,20. , 20-Trifluoro-3,6,9,12,15,18-hexaoxaicosan 3.78 g (90%) was obtained as a colorless oil.

(3) 1- (4'-p-toluenesulfonyloxy) -20,20,20-trifluoro-3,6,9,12,15,18-hexaoxaicosan obtained in the above step (2). A mixture of 1.73 g (3.34 mmol) in DMF (7 mL) with 0.66 g (6.73 mmol) ammonium bromide was heated to 80-85 ° C for 6 hours with vigorous stirring. The solvent was distilled off under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1-bromo-20,20,20-trifluoro-3,6. , 9,12,15,18-Hexaoxaicosan 1.23 g (86%) was obtained as a colorless oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.48 (2H, t, J = 6Hz), δ3.6-3.9 (22H), δ3.91 (2H) , q, J = 9Hz)

(4) 1-bromo-20,20,20-trifluoro-3,6,9,12,15,18-hexaoxaicosan 635 mg (1.49 mmol) and 6- obtained in the above step (3). Potassium carbonate 923 mg (6.7 mmol) and potassium iodide 25 mg (0.15 mmol) in a solution of hydroxy-2-methylbenzoxazole (III) 333 mg (2.23 mmol) in acetone (4 mL). The resulting mixture was refluxed for 10 hours with vigorous stirring. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 1: 1) and 2-methyl-6- (3', 6', 9'. , 12', 15', 18'-hexaoxa-20', 20', 20'-trifluoroicosanyloxy) benzoxazole 624 mg (85%) was obtained as an oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ2.60 (3H, s), δ3.6-3.8 (20H), δ3.88 (2H, m), δ3.91 (2H, q, J = 9Hz), δ4.16 (2H, m), δ6.91 (1H, dd, J = 2.1Hz, 8.6Hz), δ7.03 (1H, d, J = 2.1) Hz), δ7.50 (1H, d, J = 8.6Hz)

(5) 2-Methyl-6- (3', 6', 9', 12', 15', 18'-hexaoxa-20', 20', 20'-trifluoroico obtained in the above step (4) In a solution of 333 mg (0.673 mmol) of sanyloxy) benzoxazol and 236 mg (1.35 mmol) of 4-dimethylaminocinnamaldehyde in DMF (2.7 mL), potassium t-butoxide 224 mg (224 mg) was added to the solution while ice-cooling. 2 mmol) was added in small portions. After the addition was completed, the mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with ethyl acetate, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate: hexane = 2: 1) and 1- [6- (3', 6', 9', 12', 15', 18'-hexaoxa-20', 20', 20'-trifluoroicosanyloxy) benzoxazole-2-yl] -4- (4'-dimethylaminophenyl) pig-1,3 -Dione 267 mg (61%) was obtained as a dark red oil (mixture of geometric isomers).
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.01, 3.02, 3.03 (singlet, 6H in total), δ3.6-3.8 (20H), δ3. 8-4.0 (4H), δ4.1-4.2 (2H), δ6.4-7.1 (7H), δ7.3-7.6 (4H)

[Synthesis Example 4] 1- [6- (3', 6', 9', 12', 15', 18', 21'-Heptaoxa-23', 23', 23'-trifluorotricosanyloxy) Synthesis of benzoxazole-2-yl] -4- (4'-dimethylaminophenyl) porcine-1,3-diene (Compound 4)

(1) Weigh 0.42 g (10.5 mmol) of 60% sodium hydride in a flask in a nitrogen stream, and while cooling with ice, 1.58 g (11) of 2- (2', 2', 2'-trifluoroethoxy) ethanol. A solution of mmol) in THF (12 mL) was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 1 hour. The reaction mixture was ice-cooled again and 1- (2H-tetrahydropyran-2-yloxy) -17- (4'-p-toluenesulfonyloxy) -3,6,9,12,15-pentaoxaheptadecane 4.68 g. A solution of (9 mmol) in THF (8 mL) was added dropwise, and after completion of the addition, the reaction solution was stirred at room temperature for 13 hours. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1- (2H-tetrahydropyran-2-yloxy) -23,23. , 23-Trifluoro-3,6,9,12,15,18,21-heptaoxatricosan 4.22 g (77%) was obtained as a colorless oil.

(2) 1- (2H-tetrahydropyran-2-yloxy) -23,23,23-trifluoro-3,6,9,12,15,18,21-heptaoxa obtained in the above step (1). 1M-hydrochloric acid (0.1 mL) was added to a solution of 4.22 g (8.6 mmol) of trichosan in ethanol (40 mL), and the mixture was allowed to stand at room temperature for 22 hours. Saturated aqueous sodium hydrogen carbonate solution was added for neutralization, and the precipitated precipitate was filtered off. The filtrate was concentrated to dryness under reduced pressure, and the obtained residue was extracted with dichloromethane, and the extract was dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to dissolve the obtained residue (3.50 g) in dichloromethane (20 mL), 2.1 g (11 mmol) of p-toluenesulfonyl chloride was added, and then 1.8 mL of triethylamine (1.8 mL) was cooled with ice. 13 mmol) was added in small portions. The reaction mixture was stirred at room temperature for 24 hours, diluted with dichloromethane, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1- (4'-p-toluenesulfonyloxy) -23,23. , 23-Trifluoro-3,6,9,12,15,18,21-heptaoxatricosan 4.28 g (89%) was obtained as a colorless oil.

(3) 1- (4'-p-toluenesulfonyloxy) -23,23,23-trifluoro-3,6,9,12,15,18,21-heptaoxa obtained in the above step (2). A mixture of 4.28 g (7.6 mmol) of trichosan in DMF (16 mL) with 1.5 g (15.3 mmol) of ammonium bromide was heated to 80-85 ° C for 6 hours with vigorous stirring. The solvent was distilled off under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: methanol: dichloromethane = 1: 100) and 1-bromo-23,23,23-trifluoro-3,6. , 9,12,15,18,21-Heptaoxatricosan 3.04 g (85%) was obtained as a colorless oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.48 (2H, t, J = 6Hz), δ3.6-3.9 (22H), δ3.80 (4H) , m), δ3.91 (2H, q, J = 9Hz)

(4) With 1-bromo-23,23,23-trifluoro-3,6,9,12,15,18,21-heptaoxatricosan 431 mg (0.91 mmol) obtained in the above step (3). In a solution of 164 mg (1.1 mmol) of 6-hydroxy-2-methylbenzoxazole in acetone (4 mL), 304 mg (2.2 mmol) of potassium carbonate and 17 mg (0.1 mmol) of potassium iodide were added. The resulting mixture was refluxed for 10 hours with vigorous stirring. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate) and 2-methyl-6- (3', 6', 9', 12', 15'. , 18', 21'-Heptaoxa-23', 23', 23'-trifluorotricosanyloxy) Benzoxazole 435 mg (88%) was obtained as an oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.56 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ2.60 (3H, s), δ3.6-3.8 (24H), δ3.88 (2H, m), δ3.91 (2H, q, J = 9Hz), δ4.16 (2H, m), δ6.92 (1H, dd, J = 2.3Hz, 8.6Hz), δ7.03 (1H, d, J = 2.3) Hz), δ7.50 (1H, d, J = 8.6Hz)

(5) 2-Methyl-6- (3', 6', 9', 12', 15', 18', 21'-Heptaoxa-23', 23', 23' obtained in the above step (4) -Potassium t-butoxide in a solution of 423 mg (0.78 mmol) of trifluorotricosanyloxy) benzoxazole and 320 mg (1.6 mmol) of 4-dimethylaminocinnamaldehyde in DMF (5 mL) while cooling with ice. 260 mg (2.3 mmol) was added in small portions. After the addition was completed, the reaction solution was stirred at room temperature for 4 hours. The reaction mixture was diluted with ethyl acetate, washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate) and 1- [6- (3', 6', 9', 12', 15', 18', 21'-Heptaoxa-23', 23', 23'-Trifluorotricosanyloxy) Benzoxazole-2-yl] -4- (4'-Dimethylaminophenyl) Buta-1,3-dione 264 mg (47%) was obtained as a dark red oil (mixture of geometric isomers).
¹⁹ FNmr (CDCl ₃ ) δ-75.55 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.01, 3.02, 3.03 (singlet, 6H in total), δ3.6-3.8 (24H), δ3. 8-4.0 (4H), δ4.18 (2H, m), δ6.4-7.1 (7H), δ7.3-7.6 (4H)

[Synthesis Example 5] Synthesis of 4- (6-methoxynaphth-2-yl) bilimid [1,2-a] benzimidazole (Compound 5)

(1) 1.0 g (5.0 mmol) of 2-acetyl-6-methoxynaphthalene was heated to 135 ° C. for 5 hours in dimethylformamide diethyl acetal (5.0 mL). The crystals obtained by cooling are filtered, washed with hexane, and dried with a desiccator. 3-dimethylamino-1- (6-methoxynaphth-2-yl) prop-2-en-1-one 1.21 g (95%) ) Was obtained (mp143-144 ℃).
¹ HNmr (CDCl ₃ ) δ2.97 (3H, br.s), δ3.14 (3H, br.s), δ3.93 (3H, s), δ5.86 (1H, d, J = 12.4Hz) , δ7.1-7.2 (2H), δ7.74 (1H, d, J = 8.8Hz), δ7.82 (1H, d, J = 8.8Hz), δ7.85 (1H, d, J = 12.4Hz) ), δ7.99 (1H, dd, J = 2.0Hz, 8.8Hz), δ8.33 (1H, d, J = 2.0Hz)

(2) Diethylene glycol containing 3-dimethylamino-1- (6-methoxynaphth-2-yl) prop-2-en-1-one 370 mg (1.45 mmol) and 2-aminobenzimidazole 193 mg (1.45 mmol). It was heated to 160 ° C. for 6 hours in dimethyl ether (1.5 mL). The precipitated precipitate was collected by filtration and recrystallized from methanol to give 4- (6-methoxynaphth-2-yl) pyrimido [1,2-a] benzimidazole 117 mg (25%) (mp> 200 ° C.). ).
¹ HNmr (CDCl ₃ ) δ4.00 (3H, s), δ6.79 (1H, d, J = 8.8Hz), δ6.84 (1H, d, J = 4.4Hz), δ6.99 (1H, t) , J = 7.2Hz), δ7.25-7.35 (2H), δ7.46 (1H, t, J = 7.2Hz), δ7.58 (1H, dd, J = 2.0Hz, 8.4Hz), δ7.85 (1H, d, J = 9.2Hz), δ7.97 (1H, d, J = 8.8Hz), δ8.00 (1H, d, J = 9.2Hz), δ8.04 (1H, d, J = 2.0) Hz), δ8.82 (1H, d, J = 4.4Hz)

The synthetic route for 4- (6-methoxynaphth-2-yl) pyrimid [1,2-a] benzimidazole is shown below.

[Synthesis Example 6] 4- [6- (3', 6', 9', 12', 15', 18'-Hexaoxa-20', 20', 20'-Trifluoroeicosanyloxy) Naft-2 -Il] Pyrimid [1,2-a] Synthesis of benzimidazole (Compound 6)

(1) 2-Acetyl-6-hydroxynaphthalene 542 mg (2.91 mmol) and 1-bromo-3,6,9,12,15,18-hexaoxa-20,20,20-trifluoroicosan 957 mg (2.24 mmol) ) Was dissolved in acetone (20 mL), and the mixture obtained by adding 800 mg (5.80 mmol) of potassium carbonate and 50 mg (0.3 mmol) of potassium iodide was refluxed for 12 hours with vigorous stirring. .. The reaction mixture was concentrated to dryness under reduced pressure, and the obtained residue was extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate) and 2-acetyl-6- (3', 6', 9', 12', 15'. , 18'-Hexaoxa-20', 20', 20'-trifluoroeicosanyloxy) Naphthalene 1.10 g (92%) was obtained as a colorless oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.54 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ2.69 (3H, s), δ3.6-3.8 (20H), δ3.89 (2H, q, J) = 9Hz), δ3.93 (2H, t, J = 4.8Hz), δ4.27 (2H, t, J = 4.8Hz), δ7.15 (1H, d, J = 2.8Hz), δ7.23 ( 1H, dd, J = 2.8Hz, 9.2Hz), δ7.74 (1H, d, J = 8.4Hz), δ7.84 (1H, d, J = 8.4Hz), δ7.99 (1H, dd, J) = 1.6Hz, 8.8Hz), δ8.38 (1H, d, J = 1.6Hz)

(2) 2-Acetyl-6- (3', 6', 9', 12', 15', 18'-Hexaoxa-20', 20', 20'-trifluoroeicosanyloxy) Naphthalene 506 mg ( 0.95 mmol) was heated to 135 ° C. for 5 hours in dimethylformamide diethyl acetal (1.1 ml). The reaction mixture was cooled to room temperature and extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (eluent: ethyl acetate), and 3-dimethylamino-1- [6- (3', 6', 9', 12'). ', 15', 18'-Hexaoxa-20', 20', 20'-Trifluoroeicosanyloxy) Naft-2-yl] Prop-2-en-1-one 480 mg (85%) pale yellow Obtained as an oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.54 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.00 (3H, br.s), δ3.13 (3H, br.s), δ3.6-3.8 (20H), δ3.88 (2H, q, J = 9Hz), δ3.92 (2H, t, J = 4.8Hz), δ4.25 (2H, t, J = 4.8Hz), δ5.85 (1H) , d, J = 12Hz), δ7.14 (1H, d, J = 2.0Hz), δ7.18 (1H, dd, J = 2.0Hz, 8.8Hz), δ7.72 (1H, d J = 8.8Hz) ), δ7.82 (1H, d, J = 8.8Hz), δ7.85 (1H, d, J = 12Hz), δ7.97 (1H, dd, J = 1.6Hz, 8.8Hz), δ8.32 ( 1H, d, J = 1.6Hz)

(3) 3-dimethylamino-1- [6- (3', 6', 9', 12', 15', 18'-hexoxa-20', 20', 20'-trifluoroeicosanyloxy) Naft-2-yl] prop-2-en-1-one (390 mg (0.73 mmol)) and benzimidazole (116 mg (0.88 mmol)) were heated to 150 ° C. for 9 hours in diethylene glycol dimethyl ether (1.1 mL). The reaction mixture was cooled to room temperature and extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, the obtained residue was dissolved in ethanol, 2M-hydrochloric acid was added to adjust the pH to 2, and then the mixture was concentrated to dryness. The residue was dissolved in water, washed with diethyl ether, neutralized by adding potassium hydrogen carbonate, and extracted with ethyl acetate. The extract was washed with a small amount of water and saturated brine, and then dried over anhydrous magnesium sulfate. When the solvent is distilled off under reduced pressure, 4- [6- (3', 6', 9', 12', 15', 18'-hexoxa-20', 20', 20'-trifluoroeicosanyl) naphtho -2-yl] Pyrimid [1,2-a] benzimidazole 140 mg (31%) was obtained as a yellow oil.
¹⁹ FNmr (CDCl ₃ ) δ-75.53 (t, J = 9Hz), ¹ HNmr (CDCl ₃ ) δ3.6-3.8 (20H), δ3.89 (2H, q, J = 9Hz), δ3.97 (2H) , t, J = 4.8Hz), δ4.32 (2H, t, J = 4.8Hz), δ6.78 (1H, d, J = 8.4Hz), δ6.82 (1H, d, J = 4.0Hz) , δ6.98 (1H, t, J = 8.0Hz), δ7.29 (1H, d, J = 2.4Hz), δ7.33 (1H, dd, J = 2.4Hz, 9.2Hz), δ7.45 ( 1H, t, J = 8.4Hz), δ7.56 (1H, dd, J = 1.6Hz, 8.4Hz), δ7.84 (1H, d, J = 9.2Hz), δ7.95 (1H, d, J) = 8.8Hz), 7.99 (1H, d, J = 8.0Hz), δ8.03 (1H, d, J = 1.6Hz), δ8.82 (1H, d, J = 4.0Hz)

4- [6- (3', 6', 9', 12', 15', 18'-hexoxa-20', 20', 20'-trifluoroeicosanyl) naphth-2-yl] pyrimid [1 , 2-a] The synthetic route of benzimidazole is shown below.

[Test Example 1] Verification of the binding ability of compound to changes in human neurofibrils After dissolving compound 1 in DMSO to prepare a 10 mM solution, weigh 30 μL, add 50% ethanol, and perform a 3 mL test. It was prepared as a solution (chemical solution concentration: 100 μM). After deparaffinizing a postmortem brain tissue fixation specimen (4 μm thick) of a patient with Alzheimer's disease, 0.1% twice in 0.25% potassium permanganate solution for 20 minutes and in 10 mM phosphate buffered saline (PBS) for 2 minutes. Soaked in potassium pyrosulfite solution for 5 seconds and in 0.15% oxalic acid for 5 seconds. After washing with PBS 3 times for 2 minutes, it was immersed in the test solution for 10 minutes. After soaking in distilled water for 2 seconds, the mixture was air-dried, covered with a cover glass, and the binding of the compound to the neurofibrillary tangle was observed with a fluorescence microscope (BX61, Olympus Corporation). For the negative control, the same treatment was performed using 50% ethanol containing 1% DMSO instead of the test solution.

FIG. 1 shows a fluorescence observation image of compound 1. The fluorescence of compound 1 was observed in the neurofibrillary tangle (arrowhead), indicating that compound 1 binds to the neurofibrillary tangle. In the negative control, no neurofibrillary tangle-specific fluorescence was observed.

[Test Example 2] Verification of the binding ability of the compound to tau lesions (phosphorylated tau aggregates) After dissolving compound 1-4 in DMSO to prepare a 10 mM solution, weigh 3 μL and add 50% ethanol. In addition, 3 mL of the test solution was prepared (drug concentration: 100 μM). The test solution was immersed in a brain section of a tau genetically modified mouse (rTg4510 mouse) for 90 minutes. Then, after immersing the compound in 80% ethanol for 1 minute and in distilled water for 3 minutes, the compound was air-dried, covered with a cover glass, and the fluorescence of the compound was observed with a fluorescence microscope (Bz-8000, KEYENCE CORPORATION). For the negative control, the same treatment was performed using 50% ethanol containing 1% DMSO instead of the test solution.

Some were soaked in the test solution, washed 3 times for 10 minutes with 0.1 M phosphate buffered saline (PBST) containing 0.3% Triton X-100, and soaked in PBST containing 2% bovine serum albumin for 30 minutes. The anti-phosphorylated tau antibody (AT8, Thermo Fisher Scientific) was then diluted 1000-fold with PBST containing 0.2% bovine serum albumin and reacted with the sections overnight. After washing with PBST three times for 10 minutes, the reaction was carried out with Alexa555-labeled donkey anti-mouse IgG antibody (Thermo Fisher Scientific; 500-fold diluted) for 4 hours. After washing with PBST three times for 10 minutes, a cover glass was covered, and the binding of the compound to the tau lesion (phosphorylated tau aggregate) was observed with a fluorescence microscope (Bz-8000, KEYENCE Co., Ltd.).

FIG. 2 shows the fluorescence observation results of Compound 1-4. A and B are fluorescence observation images in the same field of view, A is the fluorescence (arrowhead) of compound 1 observed using the GFP-B filter, and B is the AT8-positive phosphorylated tau observed using the Texas Red filter (A). Arrowhead) is shown. The fluorescence of compound 1 was consistent with AT8-positive phosphorylated tau, indicating that compound 1 binds to tau lesions (phosphorylated tau aggregates) in rTg4510 mice. Similarly, in compound 2-4, fluorescence was observed in tau lesions in rTg4510 mice (arrowhead), suggesting that these compounds also bind to tau lesions.

[Test Example 3] Detection of tau lesions by fluoromagnetic resonance imaging (MRI) Compound 2 was dissolved in 20% cremofol-containing physiological saline to prepare 10 mg / mL as the administration solution. This administration solution was administered to tau gene-modified mice (rTg4510 mice) and wild-type mice under anesthesia with pentobarbital sodium (50 mg / kg, ip) from the tail vein (dose 200 mg / kg, administration rate 0.2 mL). / kg/min). After the administration was completed, MRI of the mouse head was measured using a 7 Tesla MRI device (Agilent Technologies). Fluorine MRI images were acquired by the chemical shift imaging method (CSI method).

FIG. 3 shows the measurement results. A and B show head ¹ H MRI images (A) and fluoride MRI images (B) of wild-type mice, and C and D show head ¹ H MRI images (C) and fluoride of rTg4510 mice. MRI image (D) is shown. The white lines B and D show the outline of the brain. Comparing fluoride MRI images, rTg4510 mice detected stronger MR signals in the forebrain region than wild-type mice. On the other hand, strong signals were observed around the olfactory bulb and cerebellum in both rTg4510 and wild-type mice.

[Test Example 4] Detection of tau lesions by fluoromagnetic resonance imaging (MRI) Compound 3 was dissolved in 20% cremofol-containing physiological saline to prepare 10 mg / mL as the administration solution. This administration solution was administered to tau gene-modified mice (rTg4510 mice) and wild-type mice under anesthesia with pentobarbital sodium (50 mg / kg, ip) from the tail vein (dose 200 mg / kg, administration rate 0.2 mL). / kg/min). After the administration was completed, MRI of the mouse head was measured using a 7 Tesla MRI device (Agilent Technologies). Fluorine MRI images were acquired by the chemical shift imaging method (CSI method).

FIG. 4 shows the measurement results. A and B show head ¹ H MRI images (A) and fluoride MRI images (B) of wild-type mice, and C and D show head ¹ H MRI images (C) and fluoride of rTg4510 mice. MRI image (D) is shown. The white lines B and D show the outline of the brain. Comparing fluorine MRI images, a stronger MR signal was detected in the head of rTg4510 mice than in wild-type mice, mainly in the forebrain region.

[Test Example 5] Verification that the compound crosses the blood-brain barrier and binds to tau lesions in the brain The brain was removed from the mice measured by MRI in Test Examples 3 and 4, and 0.1 M phosphate buffer containing 4% paraformaldehyde was buffered. After fixing with the solution for 24 hours, it was immersed in 0.1 M phosphate buffer containing 15% sucrose for at least 2 days. Then, a 20 μm thick section was prepared with a cryostat. This section was placed on a slide glass, air-dried, covered with a cover glass, and the fluorescence of the compound was observed with a fluorescence microscope (Bz-8000, KEYENCE CORPORATION).

FIG. 5 shows the fluorescence observation results in the hippocampus (A, C) and cerebral cortex (B, D) of rTg4510 mice treated with Compound 2 (A, B) or Compound 3 (C, D). Fluorescence (arrowheads) of the compound was observed in lesions formed in the hippocampus and cerebral cortex, indicating that the administered compound crosses the blood-brain barrier, reaches the brain, and binds to tau lesions.

[Test Example 6] Verification of the binding ability of the compound to changes in human neurofibrils After dissolving compound 5-6 in DMSO to prepare a 5 mM solution, weigh 60 μL, add 50% ethanol, and add 3 mL. (Chemical solution concentration: 100 μM). After deparaffinizing a postmortem brain tissue fixation specimen (4 μm thick) of a patient with Alzheimer's disease, 0.1% twice in 0.25% potassium permanganate solution for 20 minutes and in 10 mM phosphate buffered saline (PBS) for 2 minutes. Soaked in potassium pyrosulfite solution for 5 seconds and in 0.15% oxalic acid for 5 seconds. After washing with PBS 3 times for 2 minutes, it was immersed in the test solution for 10 minutes. After soaking in distilled water for 2 seconds, the mixture was air-dried, covered with a cover glass, and the binding of the compound to the neurofibrillary tangle was observed with a fluorescence microscope (BX61, Olympus Corporation). For the negative control, the same treatment was performed using 50% ethanol containing 1% DMSO instead of the test solution.

FIG. 6 shows fluorescence observation images of compound 5 (A) and compound 6 (B). Fluorescence of the compounds was observed in neurofibrillary tangles (arrowheads), indicating that these compounds bind to neurofibrillary tangles. No neurofibrillary tangle-specific fluorescence was observed in the negative control.

[Test Example 7] Detection of tau lesions by fluoromagnetic resonance imaging (MRI) Compound 6 was dissolved in 20% cremofol-containing physiological saline to prepare 10 mg / mL as the administration solution. This administration solution was administered from the tail vein to tau gene-modified mice (rTg4510 mice) under anesthesia with pentobarbital sodium (50 mg / kg, ip) (dose 100 mg / kg, administration rate 0.2 mL / kg / min). ). After the administration was completed, MRI of the mouse head was measured using a 7 Tesla MRI device (Agilent Technologies). Fluorine MRI images were acquired by the chemical shift imaging method (CSI method).

FIG. 7 shows the measurement results. A and B show ¹ H MRI images (A) and fluorine MRI images (B) of the sagittal section of the head. The fluorine MR signal of compound 6 was detected in the brain.

[Test Example 8] Verification that the compound crosses the blood-brain barrier and binds to tau lesions in the brain The brain was removed from the mice measured by MRI in Test Example 7 and was used with 0.1 M phosphate buffer containing 4% paraformaldehyde. After fixing for 24 hours, it was immersed in 0.1 M phosphate buffer containing 15% sucrose for at least 2 days. Then, a 20 μm thick section was prepared with a cryostat. This section was placed on a slide glass, air-dried, covered with a cover glass, and the fluorescence of the compound was observed with a fluorescence microscope (BX61, Olympus Corporation). In addition, immunohistochemical analysis with anti-phosphorylated tau antibody was performed to identify the site of tau lesion formation.

FIG. 8 shows the fluorescence observation results in the hippocampus of rTg4510 mice to which Compound 6 was administered. A and B are fluorescence observation images in the same field of view, A shows the fluorescence of compound 6, and B shows the fluorescence by the phosphorylated tau antibody. Fluorescence of compound 6 was observed in the hippocampus. Fluorescence by the phosphorylated tau antibody was also observed in the hippocampus, suggesting that the administered compound crossed the blood-brain barrier and reached the brain, binding to tau lesions.

Claims (9)

A compound or a salt thereof selected from the aniline derivative represented by the formula (1) and the naphthalene derivative represented by the formula (2).

(In the formula, R ¹ and R ² independently represent a hydrogen atom or an alkyl, respectively.
One of A ¹ and A ² indicates a hydrogen atom, and the other is a fluorine atom-substituted or unsubstituted alkoxy or

Is shown. Here, R ³ indicates a fluorine atom-substituted or unsubstituted ethylene group, R ⁴ indicates a fluorine atom substituted or unsubstituted ethyl, and n indicates an integer of 1 to 10. If n represents an integer greater than or equal to 2, R ³ may be the same or different. ) One of A ¹ and A ² indicates a hydrogen atom, and the other indicates a hydrogen atom.

The compound or salt thereof according to claim 1, wherein n indicates an integer of 5 to 8. The labeled compound according to claim 1 or a salt thereof. The compound or salt thereof according to claim 3, wherein the label is a radionuclide. An MRI diagnostic imaging agent for tauopathy containing the compound according to claim 1 or 2 or a salt thereof as an active ingredient. A diagnostic imaging agent for positron emission tomography (PET) of tauopathy containing the compound according to claim 1, 3 or 4 or a salt thereof as an active ingredient. The diagnostic imaging agent according to claim 5 or 6, wherein the tauopathy is Alzheimer's disease, progressive supranuclear palsy, corticobasal degeneration, frontotemporal dementia or Pick disease. An in vitro diagnostic agent for diagnosing tauopathy containing the compound according to any one of claims 1 to 4 or a salt thereof as an active ingredient. The in vitro diagnostic agent according to claim 8, wherein the tauopathy is Alzheimer's disease, progressive supranuclear palsy, corticobasal degeneration, frontotemporal dementia or Pick disease.

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