JP5861182B2 - Flavanol derivative-acetone derivative adduct, production method thereof, amyloid β protein aggregation inhibitor and Alzheimer preventive or therapeutic agent using the same - Google Patents

Description

  The present invention relates to a novel flavanol derivative-acetone derivative adduct. Specifically, the present invention relates to a flavanol derivative-acetone derivative adduct having an excellent amyloid β protein aggregation inhibitory action.

  Alzheimer's disease is a type of dementia whose main symptoms are cognitive decline and personality changes, such as memory impairment, disorientation, learning disabilities, attention disabilities, spatial cognitive impairment, and problem-solving disabilities. Gradually progresses and interferes with daily life. When the severity increases and the patient becomes more advanced, they cannot eat, change clothes or communicate, and eventually become bedridden.

  Currently, cholinesterase inhibitors and NMDA receptor antagonists are used as therapeutic agents for Alzheimer's disease for the purpose of reducing neuronal damage in patients. Alzheimer's major pathological change is the formation of senile plaques. In recent years, research has been conducted on drugs targeting β- and γ-secretases involved in the production of amyloid β protein, which is a constituent protein. Yes.

  On the other hand, catechins are a kind of flavonoids contained in the astringent components of tea and have various physiological activities such as blood pressure elevation inhibiting action, blood cholesterol regulating action, blood sugar level regulating action, antioxidant action, etc. It has been known. Regarding such flavonoids and derivatives thereof, prenyl flavonols having a prenyl group introduced at the C-8 position are known to have enzyme inhibitory activities such as secretase and aldose reductase (see Non-Patent Document 1). It is not known about the action of inhibiting the aggregation of amyloid β protein.

Jung HA et al., "Antidiabetic complications and anti-Alzheimer activities of sophoflavescenol, a prenylated flavonol from Sophora flavescens, and its structure-activity relationship.", Phytother Res., 2011, 25 (5), 709

  The present invention has been made in view of the above-described problems of the prior art, and an object thereof is to provide a novel flavanol derivative-acetone derivative adduct.

  Another object of the present invention is to provide a novel agent for preventing or treating Alzheimer's disease that can remarkably inhibit the aggregation of amyloid β protein in the brain.

  The above object of the present invention is achieved by the following means.

（式中、Ｒ^１〜Ｒ^８は、それぞれ独立して水素原子又は置換基を有してもよい水酸基、アミノ基若しくはチオール基であり、Ｒ^９及びＲ^１０は、それぞれ独立して水素原子若しくは置換基を有してもよい水酸基、アミノ基若しくはチオール基、又は一緒になって酸素原子であり、波線を伴う実線で表された結合（ａ）は、単結合又は二重結合であり、Ｒ^１１及びＲ^１２は、それぞれ独立して水素原子又は一般式（II）

（式中、Ｒ^１３及びＲ^１４は、それぞれ独立して水素原子又は炭素数１〜５のアルキル基であり、波線を伴う実線で表された結合（ｂ）〜（ｄ）は、何れか１つが二重結合で残りが単結合であり、ｎは１〜４の整数である）で示されるアルケニル基であり、かつＲ^１１とＲ^１２が同時に水素原子になることはない）で示される、フラバノール誘導体−アセトン誘導体付加物である。 (1) That is, the present invention relates to the general formula (I)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or The bond (a) which is a hydroxyl group, an amino group or a thiol group which may have a substituent, or an oxygen atom together and is represented by a solid line with a wavy line is a single bond or a double bond, and R ¹¹ and R ¹² are each independently a hydrogen atom or the general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the remaining is a single bond, and n is an integer of 1 to 4, and R ¹¹ and R ¹² do not simultaneously become hydrogen atoms). A flavanol derivative-acetone derivative adduct.

(2) In the present invention, R ^{1 to} R ^{8 in} the general formula (I) are each independently a hydrogen atom or a hydroxyl group which may have a substituent, and R ⁹ and R ¹⁰ are a hydrogen atom. The flavanol derivative-acetone derivative adduct according to (1), wherein the bond (a) is a single bond.

  (3) The present invention also relates to (1) or (2), wherein the bonds (b) and (d) in the general formula (II) are single bonds and the bond (c) is a double bond. A flavanol derivative-acetone derivative adduct.

  (4) The present invention is also obtained by reacting catechins selected from the group consisting of catechin, epicatechin, gallocatechin and epigallocatechin or a derivative thereof with prenylacetone or a derivative thereof, The flavanol derivative-acetone derivative adduct according to any one of (1) to (3).

  (5) The present invention also provides 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3-b] [1] benzopyran -2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnona-3,7-dienyl) [2] Benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4 , 8,12-trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol or 6aS, 12aR-5,6a , 7, 1 a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenyl) [2] benzopyrano [4,3- b] [1] The flavanol derivative-acetone derivative adduct according to any one of (1) to (4), which is benzopyran-2,3,8,10-tetrol.

（式中、Ｒ^１〜Ｒ^８は、それぞれ独立して水素原子又は置換基を有してもよい水酸基、アミノ基若しくはチオール基であり、Ｒ^９及びＲ^１０は、それぞれ独立して水素原子若しくは置換基を有してもよい水酸基、アミノ基若しくはチオール基、又は一緒になって酸素原子であり、波線を伴う実線で表された結合（ａ）は、単結合又は二重結合である）で示されるフラバノール誘導体と、一般式（IV）

（式中、Ｒ^１１及びＲ^１２は、それぞれ独立して水素原子又は一般式（II）

（式中、Ｒ^１３及びＲ^１４は、それぞれ独立して水素原子又は炭素数１〜５のアルキル基であり、波線を伴う実線で表された結合（ｂ）〜（ｄ）は、何れか１つが二重結合で残りが単結合であり、ｎは１〜４の整数である）で示されるアルケニル基であり、かつＲ^１１とＲ^１２が同時に水素原子になることはない）で示されるアセトン誘導体と、を反応させることを特徴とする、一般式（Ｉ）

（式中、Ｒ^１〜Ｒ^１２及び結合（ａ）は、前記定義通りである）で示されるフラバノール誘導体−アセトン誘導体付加物の製造方法である。 (6) Furthermore, the present invention provides a compound of the general formula (III)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or A hydroxyl group, an amino group or a thiol group, which may have a substituent, or an oxygen atom together, and a bond (a) represented by a solid line with a wavy line is a single bond or a double bond) The flavanol derivatives shown and the general formula (IV)

(Wherein R ¹¹ and R ¹² are each independently a hydrogen atom or a general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the rest is a single bond, n is an integer of 1 to 4), and R ¹¹ and R ¹² do not simultaneously become a hydrogen atom) A derivative of the compound represented by the general formula (I)

(Wherein R ^{1 to} R ¹² and the bond (a) are as defined above), a flavanol derivative-acetone derivative adduct production method.

(7) In the present invention, R ^{1 to} R ^{8 in} the general formula (III) are each independently a hydrogen atom or a hydroxyl group which may have a substituent, and R ⁹ and R ¹⁰ are a hydrogen atom. The method for producing an flavanol derivative-acetone derivative adduct according to (6), wherein the bond (a) is a single bond.

  (8) The present invention also relates to (6) or (7), wherein the bonds (b) and (d) in the general formula (II) are single bonds and the bond (c) is a double bond. It is a manufacturing method of a flavanol derivative-acetone derivative adduct.

  (9) In the present invention, the flavanol derivative is a catechin selected from the group consisting of catechin, epicatechin, gallocatechin and epigallocatechin, or a derivative thereof, any one of (6) to (8) The flavanol derivative-acetone derivative adduct described in 1. above.

  (10) The present invention is also the method for producing a flavanol derivative-acetone derivative adduct according to any one of (6) to (9), wherein the acetone derivative is prenylacetone or a derivative thereof.

（式中、Ｒ^１〜Ｒ^８は、それぞれ独立して水素原子又は置換基を有してもよい水酸基、アミノ基若しくはチオール基であり、Ｒ^９及びＲ^１０は、それぞれ独立して水素原子若しくは置換基を有してもよい水酸基、アミノ基若しくはチオール基、又は一緒になって酸素原子であり、波線を伴う実線で表された結合（ａ）は、単結合又は二重結合であり、Ｒ^１１及びＲ^１２は、それぞれ独立して水素原子又は一般式（II）

（式中、Ｒ^１３及びＲ^１４は、それぞれ独立して水素原子又は炭素数１〜５のアルキル基であり、波線を伴う実線で表された結合（ｂ）〜（ｄ）は、何れか１つが二重結合で残りが単結合であり、ｎは１〜４の整数である）で示されるアルケニル基であり、かつＲ^１１とＲ^１２が同時に水素原子になることはない）で示されるフラバノール誘導体−アセトン誘導体付加物又はその生理学的に許容される塩を有効成分とする、アミロイドβ蛋白凝集阻害剤である。 (11) Furthermore, the present invention provides a compound of the general formula (I)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or The bond (a) which is a hydroxyl group, an amino group or a thiol group which may have a substituent, or an oxygen atom together and is represented by a solid line with a wavy line is a single bond or a double bond, and R ¹¹ and R ¹² are each independently a hydrogen atom or the general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the rest is a single bond, n is an integer of 1 to 4, and R ¹¹ and R ¹² are not hydrogen atoms at the same time). Derivative-acetone derivative adduct or a physiologically acceptable salt thereof as an active ingredient is an amyloid β protein aggregation inhibitor.

(12) In the present invention, the flavanol derivative-acetone derivative adduct is a hydroxyl group in which R ^{1 to} R ^{8 in} the general formula (I) may independently have a hydrogen atom or a substituent, The amyloid β protein aggregation inhibitor according to (11), wherein R ⁹ and R ¹⁰ are hydrogen atoms, and the bond (a) is a single bond.

  (13) The present invention is also directed to the flavanol derivative-acetone derivative adduct, wherein the bonds (b) and (d) of the general formula (II) are single bonds, and the bond (c) is a double bond. The amyloid β protein aggregation inhibitor according to (11) or (12).

  (14) In the present invention, the flavanol derivative-acetone derivative adduct is obtained by reacting catechins selected from the group consisting of catechin, epicatechin, gallocatechin, and epigallocatechin or a derivative thereof with prenylacetone or a derivative thereof. It is obtained by making it an amyloid beta protein aggregation inhibitor as described in any one of (11)-(13) characterized by the above-mentioned.

(15) The present invention also provides that the flavanol derivative-acetone derivative adduct is 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [ 4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnona -3,7-dienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12-trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10 − Trol or 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11,15 -Tetraenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, the amyloid β protein aggregation according to any one of (11) to (14) An inhibitor .

  (16) Furthermore, the present invention is an agent for preventing or treating Alzheimer's disease comprising the amyloid β protein aggregation inhibitor according to any one of (11) to (15).

  Since the flavanol derivative-acetone derivative adduct of the present invention inhibits aggregation of amyloid β protein produced in the brain, it can remarkably suppress the generation of senile plaques due to aggregation of amyloid β protein.

  Moreover, since the flavanol derivative-acetone derivative adduct of the present invention has an excellent radical scavenging ability, it can remarkably erase active oxygen generated from senile plaques.

  Furthermore, since the flavanol derivative-acetone derivative adduct of the present invention is excellent in the permeability to the brain blood barrier, it can easily reach the brain even by systemic administration by oral or injection.

  Therefore, when the flavanol derivative-acetone derivative adduct of the invention is used as an agent for preventing or treating Alzheimer's disease, the onset and progression of Alzheimer's disease can be remarkably suppressed or delayed.

（式中、Ｒ^１〜Ｒ^８は、それぞれ独立して水素原子又は置換基を有してもよい水酸基、アミノ基若しくはチオール基であり、Ｒ^９及びＲ^１０は、それぞれ独立して水素原子若しくは置換基を有してもよい水酸基、アミノ基若しくはチオール基、又は一緒になって酸素原子であり、波線を伴う実線で表された結合（ａ）は、単結合又は二重結合であり、Ｒ^１１及びＲ^１２は、それぞれ独立して水素原子又は一般式（II）

（式中、Ｒ^１３及びＲ^１４は、それぞれ独立して水素原子又は炭素数１〜５のアルキル基であり、波線を伴う実線で表された結合（ｂ）〜（ｄ）は、何れか１つが二重結合で残りが単結合であり、ｎは１〜４の整数である）で示されるアルケニル基であり、かつＲ^１１とＲ^１２が同時に水素原子になることはない）で示されるものである。 The flavanol derivative-acetone derivative adduct of the present invention (hereinafter referred to as “flavanol derivative-acetone derivative adduct (I)”) has the following general formula (I):

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or The bond (a) which is a hydroxyl group, an amino group or a thiol group which may have a substituent, or an oxygen atom together and is represented by a solid line with a wavy line is a single bond or a double bond, and R ¹¹ and R ¹² are each independently a hydrogen atom or the general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the rest is a single bond, and n is an integer of 1 to 4, and R ¹¹ and R ¹² do not simultaneously become hydrogen atoms). It is.

In the flavanol derivative-acetone derivative adduct (I) of the present invention, the substituents for R ^{1 to} R ¹⁰ include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert- Examples thereof include a linear, branched or cyclic alkyl group having 1 to 5 carbon atoms such as a butyl group, a cyclopropyl group and a cyclobutyl group, a hydroxyl group which may have a substituent, an amino group or a thiol group.

In addition, examples of the alkyl group of R ¹³ and R ¹⁴ include carbon numbers such as methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, sec-butyl group, tert-butyl group, cyclopropyl group, and cyclobutyl group. Examples include 1 to 5 linear, branched or cyclic alkyl groups.

（式中、Ｒ^１〜Ｒ¹⁰は、前記定義通りである）で示されるフラバノール誘導体（以下「フラバノール誘導体（III）」という）と、 The flavanol derivative-acetone derivative adduct (I) of the present invention has the following general formula (III):

(Wherein R ^{1 to} R ¹⁰ are as defined above) (hereinafter referred to as “flavanol derivative (III)”);

（式中、Ｒ^１１及びＲ^１２は、前記定義通りである）で示されるアセトン誘導体（以下「アセトン誘導体（IV）」という）と、を反応させることにより得ることができる。 Formula (IV)

(Wherein R ¹¹ and R ¹² are as defined above) and can be obtained by reacting with an acetone derivative (hereinafter referred to as “acetone derivative (IV)”).

The flavanol derivative (III) used in the present invention is not particularly limited as long as it is represented by the above general formula (III). Preferably, R ^{1 to} R ⁸ are each independently a hydrogen atom. Or a hydroxyl group that may have a substituent, R ⁹ and R ¹⁰ are hydrogen atoms, and the bond (a) is a single bond, more preferably catechins or derivatives thereof, Preferably, catechin, epicatechin, gallocatechin, epigallocatechin or derivatives thereof represented by the following formulas (V) to (VIII) are preferable.

  Further, the acetone derivative (IV) used in the present invention is not particularly limited as long as it is represented by the above general formula (IV), but preferably the bond (b) of the above general formula (II) ) And (d) are single bonds and the bond (c) is preferably a double bond, and particularly preferably a dimethylallyl group (n = 1), a geranyl group (n = 2), a farnesyl group (n = 3), prenylacetone having a prenyl group such as geranylgeranyl group (n = 4) or a derivative thereof is preferable.

  The production method of the flavanol derivative-acetone derivative adduct (I) of the present invention will be specifically described. First, the flavanol derivative (III) is dissolved in an appropriate solvent such as THF, dioxane, diethyl ether, and the like. Acetone derivative (IV) is added dropwise at −5 to −30 ° C. with stirring and reacted for 1 to 24 hours, and then the reaction is stopped by adding a weakly basic substance solution such as water or sodium hydrogencarbonate. The obtained reaction mixture is extracted with an organic solvent such as ethyl acetate and diethyl ether, washed with water, evaporated to remove the solvent, and purified by silica gel column chromatography to obtain the desired product.

  Specific examples of the thus obtained flavanol derivative-acetone derivative adduct (I) of the present invention include, for example, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl) 3-pentenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- ( (E) -4,8-dimethylnona-3,7-dienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12-trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [ Benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetra Methylheptadeca-3,7,11,15-tetraenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aR, 12aR-5, 6a, 7, 12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aR, 12aR-5,6a , 7,12a-Tetrahydro-5-methyl 5-((E) -4,8-dimethylnona-3,7-dienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8 , 0-tetrol, 6aR, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12-trimethyltrideca-3,7,11-trienyl) [2 Benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aR, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E, 11E ) -4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,4, 8 10-pentol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnona-3,7-dienyl) [2] benzopyrano [4,3-b ] [1] Benzopyran-2,3,4,8,10-pentol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12- Trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,4,8,10-pentol, 6aS, 12aR-5,6a, 7, 12a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenyl) [2] benzopyrano [4,3- b] [1 Benzopyran-2,3,4,8,10-pentol, 6aR, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3 -B] [1] benzopyran-2,3,4,8,10-pentol, 6aR, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnon- 3,7-dienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,4,8,10-pentol, 6aR, 12aR-5,6a, 7,12a-tetrahydro-5 Methyl 5-((3E, 7E) -4,8,12-trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,4 8,10-pen 6aR, 12aR-5, 6a, 7, 12a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11, 15-tetraenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,4,8,10-pentol and the like.

  The flavanol derivative-acetone derivative adduct (I) of the present invention inhibits the aggregation of amyloid β protein produced in the brain, remarkably suppresses the production of senile plaques due to the aggregation of amyloid β protein, and is generated from senile plaques Since the active oxygen is also eliminated, it can be used as an amyloid β protein aggregation inhibitor and an Alzheimer's disease preventive or therapeutic agent.

  The amyloid β protein aggregation inhibitor of the present invention is an effective amount of the above flavanol derivative-acetone derivative adduct (I) alone, or a tablet, pill, powder, powder, granule, syrup, liquid, suspension. It can be orally administered to a patient as an agent, emulsion, capsule or the like. Moreover, it can be administered as an injection intravenously, intramuscularly, intradermally, subcutaneously, intraperitoneally, intraarterially, intrathecally, or the like. Among the above-mentioned, a preferable formulation form, administration form, and the like are appropriately selected by a doctor according to the patient's age, sex, constitution, symptoms, treatment timing, and the like.

  When this preparation is made into a solid preparation such as tablets, pills, powders, powders, granules, etc., the above flavanol derivative-acetone derivative adduct (I) is added to an appropriate additive such as lactose, sucrose according to a conventional method. Sugar, mannitol, corn starch, synthetic or natural gums, excipients such as crystalline cellulose, starch, cellulose derivatives, binders such as gum arabic, gelatin, polyvinylpyrrolidone, carboxymethyl cellulose calcium, carboxymethyl cellulose Mix appropriately with disintegrants such as sodium, starch, corn starch, sodium alginate, lubricants such as talc, magnesium stearate, sodium stearate, fillers or diluents such as calcium carbonate, sodium carbonate, calcium phosphate, sodium phosphate Can be manufactured. Tablets and the like may be coated with sugar coating, gelatin, enteric coating, film coating, etc. using an appropriate coating base as necessary.

  When the preparation is a liquid preparation such as injection, eye drop, nasal drop, inhalant, spray, lotion, syrup, solution, suspension, emulsion, etc., the above flavanol derivative-acetone derivative adduct (I) purified water, suitable buffer solution such as phosphate buffer, physiological saline solution, physiological salt solution such as Ringer solution, lock solution, vegetable oil such as cacao butter, sesame oil, olive oil, mineral oil, higher alcohol , Dissolved in organic solvents such as higher fatty acids and ethanol, emulsifiers such as cholesterol, suspending agents such as gum arabic, dispersion aids, wetting agents, polyoxyethylene hydrogenated castor oil, polyethylene glycol Surfactants such as sodium phosphate, solubilizers such as sodium phosphate, stabilizers such as sugar, sugar alcohol, albumin, preservatives such as parabens, sodium chloride, glucose, glycerin Isotonic agent, buffer, soothing agent, anti-adsorption agent, moisturizer, antioxidant, colorant, sweetener, flavor, fragrance, etc. It can be prepared as a suspension, liposome or emulsion. At this time, the injection preferably has a physiological pH, preferably in the range of 6-8.

  In order to make this preparation into a semi-solid preparation such as a lotion, cream or ointment, the above flavanol derivative-acetone derivative adduct (I) is added to fat, fatty oil, lanolin, petrolatum, paraffin, wax, plaster, resin. It can be produced by appropriately mixing with plastics, glycols, higher alcohols, glycerin, water, emulsifiers, suspending agents and the like.

  The content of the flavanol derivative-acetone derivative adduct (I) contained in the amyloid β protein aggregation inhibitor of the present invention varies depending on the dosage form, severity, target dose, etc. Is 10 to 80% by weight, preferably 20 to 40% by weight, based on the total weight of the preparation.

  The dosage of the amyloid β protein aggregation inhibitor of the present invention varies depending on the age, weight and symptoms of the patient, the intended dosage form and method, therapeutic effect, treatment period, etc., and the exact amount is determined by a doctor. However, in general, the above flavanol derivative-acetone derivative adduct (I) in terms of dose is 10 to 500 mg for adults per day for oral administration, and 0.5 to 0.5 for intravenous administration. 100 mg is administered in one or several divided doses.

  Next, the flavanol derivative-acetone derivative adduct of the present invention and the amyloid β protein aggregation inhibitor using the same will be described in more detail with reference to examples, but the present invention is limited to such examples. is not.

Synthesis of catechin geranylacetone adducts

  (+)-Catechin 5.00 g (17.2 mmol) was dried with a vacuum pump for 3 hours and then purged with argon, and 130 ml of dry tetrahydrofuran (THF) was slowly added and stirred. The reaction vessel was cooled to −30 ° C., 5.78 ml (25.8 mmol) of geranyl acetone and 3.74 ml (20.7 mmol) of trimethylsilyl trifluoromethanesulfonate (TMSOTf) were added to the reaction solution, and the reaction was conducted at −30 ° C. went. After 3 hours and 15 minutes, thin layer chromatography (Silica Gel F254, 0.25 mm, MERCK, toluene: acetone: methanol = 7: 3: 1) was followed by TMSOTf 1.87 ml (10.3 mmol) The mixture was further stirred for 16 hours and 15 minutes (total reaction time 19 hours and 30 minutes).

  The reaction mixture was quenched with saturated aqueous sodium hydrogen carbonate solution, poured into ice water, extracted three times with ethyl acetate, the organic layer was washed three times with saturated brine, dehydrated with anhydrous sodium sulfate, and evaporated. , 9.44 g of reaction mixture was obtained. The residue was purified with 700 g of silica gel column chromatography (Silica Gel 60, chemical, 63-200 μM, MERCK), and 6.89 g of catechingeranyl acetone adduct was added from the eluate of toluene: acetone: methanol (7: 3: 0.25) ( Yield 85.7%) was obtained.

The NMR spectrum (Varian Mercury AS-400) and Mass spectrum (JEOL JMS-SX102A, Operating Mode, EI +) of the obtained catechin geranylacetone adduct are shown below.
1H NMR (DMSO) δ: 1.35 to 1.63 (s, 12H), 1.83 to 2.10 (m, 8H), 2.29 to 2.35 (m, 1H), 2.79 to 2.82 (m, 1H), 3.65 to 3.74 (m, 1H), 4.34 (d, J = 9.0 Hz, 1H), 4.36 (d, J = 9.0 Hz, 1H) ), 4.42 (d, J = 9.0 Hz, 1H), 4.43 (d, J = 9.0 Hz, 1H), 5.01 (m, 1H), 5.03 (m, 1H), 5.03 (m, 1H), 5.16 (m, 1H), 5.79 (m, 1H), 5.94 (m, 1H), 6.48 to 6.51 (s, 1H), 6 .91 to 6.92 (s, 1H), 8.78 to 8.80 (s, 1H), 9.02 to 9.03 (s, 1H), 9.03 to 9.05 (s, 1H) , 9.29 to 9.31 (s, 1H)
_{HR-MS m / z: cald} for C 28 H 34 O 6
[M +]: 466.2355 found 466.2353

Synthesis of catechingeranylgeranylacetone adducts

  3.65 g (12.6 mmol) of (+)-catechin was dried with a vacuum pump for 2 hours, purged with argon, and slowly added dry THF (100 ml) and stirred. The reaction vessel was cooled to −50 ° C., 5.61 ml (15.1 mmol) of geranylgeranylacetone (teprenone) and 2.73 ml (15.1 mmol) of TMSOTf were added to the reaction solution, and the reaction was performed at −50 ° C. After 16 hours, thin layer chromatography (Silica Gel F254, 0.25 mm, MERCK, toluene: acetone: methanol = 7: 3: 1) was confirmed to confirm the disappearance of the raw materials.

  The reaction mixture was quenched with saturated aqueous sodium hydrogen carbonate solution, poured into ice water, extracted three times with ethyl acetate, the organic layer was washed three times with saturated brine, dehydrated with anhydrous sodium sulfate, and the solvent was removed. After leaving, a reaction mixture was obtained. The residue was purified by 600 g of silica gel column chromatography (Silica Gel 60, chemical, 63-200 μM, MERCK), and catechingeranylgeranylacetone adduct from the eluate of dichloromethane: hexane: mertanol (7: 3: 0.6) 12. 3g8 yield 67.5%) was obtained.

The NMR spectrum (Varian Mercury AS-400) and Mass spectrum (SHIMAZU LCMS-IT-TOF Operating Mode, EI-) of the obtained catechingeranylgeranylacetone adduct were shown below.
1H NMR (CD3OD) δ: 1.43 to 2.17 (m, 34H), 2.45 (dd, J = 8.0 and 15.2 Hz, 1 H), 2.96 (dd, J = 6.0 and 15.6 Hz) , 1H), 3.79 (m, 1H), 4.41 (m, 1H), 5.06 (m, 4H), 5.94 (d, J = 1.6 Hz, 1H), 5.96 ( d, J = 1.6 Hz, 1H), 6.53 (s, 1H), 7.04 (s, 1H)
HR-MS m / z: caldfor C28H34O6 [M +]: 602.3607 found 601.3574

Evaluation of aggregation inhibitory activity against amyloid β (1-42) by thioflavin T assay (ThT) method

  Aggregation inhibitory activity against Aβ42 by the ThT method of the catechingeranylacetone adducts and catechingeranylgeranylacetone adducts obtained in Examples 1 and 2 was evaluated according to the following literature.

  Hironobu Naiki, et al., "Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavine T", Analytical Biochemistry, Vol. 177, Issue 2, Pages 244-249, 1989

  Kazuma Murakami, et al., "Neurotoxicity and Physicochemical Properties of Aβ Mutant Peptides from Cerebral Amyloid Angiopathy", THE JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 278, No. 46, Pages 46179-46187, 2003

  A test compound was dissolved in 50% DMSO / 0.02% NH4OH to prepare a 2.5 mM stock solution. 1 mg of Aβ42 was dissolved in 221.5 μl of 0.02% NH 4 OH to prepare a 1.0 mM stock solution, diluted with 50 mM MPB (pH 7.4) containing 100 mM NaCl, 500 μl of 25 μM Aβ42 solution containing only Aβ42, 1% DMSO ( v / v) and 25 μM Aβ42 solution 500 μl containing 10 μM or 50 μM of the test compound and DMSO 1% (v / v) were prepared in duplicate. The blank was Aμ42 replaced with 0.02% NH4OH.

  The prepared sample was incubated at 37 ° C., and 10 μl was taken at 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 24, 48 hr, and 5 μM Th-T / 50 mM Gly-NaOH prepared in advance. (PH 8.9) In addition to 1 ml, stirring, and the fluorescence intensity of 487 nm by excitation light of 435 nm was measured with a fluorometer (Shimadzu spectrofluorometer RF-5300PC series, manufactured by Shimadzu Corporation).

  Table 1 shows the measurement results of the aggregation inhibitory activity of the test compound. For comparison, catechin geranylacetone adducts, catechins, catechins, epicatechin, epicatechin gallate, epigallocatechin and epigallocatechin gallate, and typical antioxidants myricetin, pyrogallol, ellag The measurement results of acid and α-tocopherol are also shown in Table 1. Further, the brain blood barrier permeability of these compounds was calculated with reference to the following literature, and is also shown in Table 1.

Hou.TJ et al., "ADME Evaluation in Drug Discovery. 3. Modeling Blood-Brain Barrier Partitioning Using Simple Molecular Descriptors", J. Chem. Inf. Comput. Sci., 2003, 43, 2137-2152

  As is apparent from Table 1, the catechingeranylacetone adduct and the catechingeranylgeranylacetone adduct, which are the flavanol derivative-acetone derivative adducts of the present invention, have extremely excellent amyloid β protein aggregation inhibitory activity and also have high radical scavenging activity. I found out that I have it. In addition, since the flavanol derivative-acetone derivative adduct of the present invention has excellent cerebral blood barrier permeability, it reaches the brain by systemic administration by oral or injection, and is excellent for Alzheimer's disease. Expected to have a preventive or therapeutic effect.

  As described above, the flavanol derivative-acetone derivative adduct of the present invention inhibits the aggregation of amyloid β protein produced in the brain, remarkably suppresses the formation of senile plaques due to the aggregation of amyloid β protein, and Since it also eliminates active oxygen generated from water, it is extremely useful when used as an inhibitor of amyloid β protein aggregation and as a preventive or therapeutic agent for Alzheimer's disease.

Claims (16)

Formula (I)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or The bond (a) which is a hydroxyl group, an amino group or a thiol group which may have a substituent, or an oxygen atom together and is represented by a solid line with a wavy line is a single bond or a double bond, and R ¹¹ and R ¹² are each independently a hydrogen atom or the general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the remaining is a single bond, and n is an integer of 1 to 4, and R ¹¹ and R ¹² do not simultaneously become hydrogen atoms). Flavanol derivative-acetone derivative adduct. R ^{1 to} R ^{8 in} the general formula (I) are each independently a hydrogen atom or a hydroxyl group that may have a substituent, R ⁹ and R ¹⁰ are hydrogen atoms, and the bond (a) is a single bond. The flavanol derivative-acetone derivative adduct according to claim 1, wherein   The flavanol derivative-acetone derivative adduct according to claim 1 or 2, wherein the bonds (b) and (d) of the general formula (II) are single bonds and the bond (c) is a double bond.   The catechin selected from the group consisting of catechin, epicatechin, gallocatechin and epigallocatechin or a derivative thereof and prenylacetone or a derivative thereof are obtained, and obtained by reacting any one of claims 1 to 3. The flavanol derivative-acetone derivative adduct according to item 1.   6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10 -Tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnona-3,7-dienyl) [2] benzopyrano [4,3-b] [1] Benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12-trimethyltrideca -3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol or 6aS, 12aR-5,6a, 7,12a-tetrahydro 5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenyl) [2] benzopyrano [4,3-b] [1 The flavanol derivative-acetone derivative adduct according to any one of claims 1 to 4, which is benzopyran-2,3,8,10-tetrol. General formula (III)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or A hydroxyl group, an amino group or a thiol group, which may have a substituent, or an oxygen atom together, and a bond (a) represented by a solid line with a wavy line is a single bond or a double bond) The flavanol derivative shown,
Formula (IV)

(Wherein R ¹¹ and R ¹² are each independently a hydrogen atom or a general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the rest is a single bond, n is an integer of 1 to 4), and R ¹¹ and R ¹² do not simultaneously become a hydrogen atom) A derivative;
In which general formula (I) is reacted

(Wherein R ^{1 to} R ¹² and bond (a) are as defined above)
The manufacturing method of the flavanol derivative-acetone derivative adduct shown by these. R ^{1 to} R ^{8 in} the general formula (III) are each independently a hydrogen atom or a hydroxyl group that may have a substituent, R ⁹ and R ¹⁰ are hydrogen atoms, and the bond (a) is a single bond. The manufacturing method of the flavanol derivative-acetone derivative adduct of Claim 6 which is these.   The method for producing a flavanol derivative-acetone derivative adduct according to claim 6 or 7, wherein the bonds (b) and (d) of the general formula (II) are single bonds and the bond (c) is a double bond. .   The flavanol derivative-acetone derivative adduct according to any one of claims 6 to 8, wherein the flavanol derivative is a catechin selected from the group consisting of catechin, epicatechin, gallocatechin and epigallocatechin or a derivative thereof. Manufacturing method.   The said acetone derivative is a manufacturing method of the flavanol derivative-acetone derivative adduct of any one of Claims 6-9 which is prenyl acetone or its derivative (s). Formula (I)

(Wherein R ^{1 to} R ⁸ are each independently a hydrogen atom or a hydroxyl group, amino group or thiol group which may have a substituent, and R ⁹ and R ¹⁰ are each independently a hydrogen atom or The bond (a) which is a hydroxyl group, an amino group or a thiol group which may have a substituent, or an oxygen atom together and is represented by a solid line with a wavy line is a single bond or a double bond, and R ¹¹ and R ¹² are each independently a hydrogen atom or the general formula (II)

(In the formula, R ¹³ and R ¹⁴ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and any one of the bonds (b) to (d) represented by a solid line with a wavy line is 1 One is a double bond and the rest is a single bond, n is an integer of 1 to 4, and R ¹¹ and R ¹² are not hydrogen atoms at the same time). Derivative-acetone derivative adduct or a physiologically acceptable salt thereof as an active ingredient is an amyloid β protein aggregation inhibitor. In the flavanol derivative-acetone derivative adduct, R ^{1 to} R ^{8 in} the general formula (I) are each independently a hydrogen atom or a hydroxyl group which may have a substituent, and R ⁹ and R ¹⁰ are hydrogen atoms. The amyloid β protein aggregation inhibitor according to claim 11, wherein the bond (a) is a single bond.   13. The flavanol derivative-acetone derivative adduct according to claim 11, wherein the bonds (b) and (d) of the general formula (II) are single bonds, and the bond (c) is a double bond. Amyloid β protein aggregation inhibitor.   The flavanol derivative-acetone derivative adduct is obtained by reacting a catechin selected from the group consisting of catechin, epicatechin, gallocatechin and epigallocatechin or a derivative thereof with prenylacetone or a derivative thereof. The amyloid β protein aggregation inhibitor according to any one of claims 11 to 13. The flavanol derivative-acetone derivative adduct is 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5- (4-methyl 3-pentenyl) [2] benzopyrano [4,3-b] [1. Benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((E) -4,8-dimethylnona-3,7-dienyl) [ 2] Benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol, 6aS, 12aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E) -4,8,12-trimethyltrideca-3,7,11-trienyl) [2] benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol or 6aS, 1 aR-5,6a, 7,12a-tetrahydro-5-methyl 5-((3E, 7E, 11E) -4,8,12,16-tetramethylheptadeca-3,7,11,15-tetraenyl) [ 2] The amyloid β protein aggregation inhibitor according to any one of claims 11 to 14, which is benzopyrano [4,3-b] [1] benzopyran-2,3,8,10-tetrol.   An agent for preventing or treating Alzheimer's disease comprising the amyloid β protein aggregation inhibitor according to any one of claims 11 to 15.