JP2007204380A - New compound and pharmaceutical application thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new compound useful in treating diseases related to cell motility(wandering ability), and to provide applications thereof. <P>SOLUTION: The new compound is represented by the formula A-B( wherein, A is either one group among groups(a) to (e); B is either one group among groups(f) to (j); and R<SB>1</SB>is an alkyl). <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a novel compound or a pharmacologically acceptable salt thereof, a pharmaceutical composition containing them as an active ingredient, a cell motility inhibitor, a cell infiltration inhibitor, and a cell growth inhibitor.

  A substance having an inhibitory activity on cell motility (migration ability), invasive ability, proliferation and the like is considered to be useful as a therapeutic agent for diseases related to cell motility, cell invasion, cell proliferation and the like.

Conventionally, UTKO-1 to 5 (see Patent Documents 1 and 2) and the like are known as inhibitory active substances for cell motility (migration ability), invasive ability, proliferation, and the like.
JP 2005-330265 A International Publication No. 05/102981 Pamphlet

  An object of the present invention is to provide a novel compound useful as a therapeutic agent for diseases related to cell motility (migration ability) and use thereof.

The present inventors have prepared compounds represented by the following formulas (1) to (6) (hereinafter referred to as “compound (1)”, “compound (2)”, “compound (3)”, “compound (4 ) "," Compound (5) ", and" compound (6) ") were found to have inhibitory activity on tumor cell motility (migration ability).

  In addition, the present inventors have found that the compounds (2) to (6) have an activity of inhibiting the growth of tumor cells and exhibit a strong toxicity to the tumor cells. Based on the above results, the present inventors have completed the present invention.

That is, the compound according to the present invention is a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof.

In the formula (I), A is a group represented by any of the following formulas (a) to (e), B is a group represented by any of the following formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  The compound according to the present invention is any one of the above-mentioned compounds (1) to (6) or a pharmacologically acceptable salt thereof.

The pharmaceutical composition according to the present invention contains a compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  The pharmaceutical composition according to the present invention contains any one of the above-mentioned compounds (1) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

The cell motility inhibitor according to the present invention contains the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  The cell motility inhibitor according to the present invention contains any one of the above-mentioned compounds (1) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

The agent for inhibiting cell invasion according to the present invention contains the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  In addition, the drug for inhibiting cell invasion according to the present invention contains any one of the above-mentioned compounds (2) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

The cell growth inhibitor according to the present invention contains the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  The cell growth inhibitor according to the present invention contains any one of the above-mentioned compounds (2) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

The method for inhibiting the motility of a cell according to the present invention includes a step of allowing a compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof to act. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  Moreover, the method for inhibiting the motility of a cell according to the present invention includes a step of acting any one of the above-mentioned compounds (1) to (6) or a pharmacologically acceptable salt thereof.

The method for inhibiting cell invasion according to the present invention includes a step of allowing the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof to act. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  Moreover, the method for inhibiting cell infiltration according to the present invention includes a step of allowing any one of the above-mentioned compounds (2) to (6) or a pharmacologically acceptable salt thereof to act.

The method for inhibiting cell growth according to the present invention comprises a step of allowing the compound represented by the above general formula (I) or a pharmacologically acceptable salt thereof to act. In the formula (I), A is a group represented by any one of the above formulas (a) to (e), B is a group represented by any one of the above formulas (f) to (j), R ₁ is a lower alkyl group having 1 to 4 carbon atoms.

  Moreover, the method for inhibiting cell proliferation according to the present invention includes a step of allowing any one of the above-mentioned compounds (2) to (6) or a pharmacologically acceptable salt thereof to act.

  ADVANTAGE OF THE INVENTION According to this invention, the novel compound useful as a therapeutic agent of the disease related to cell motility (migration ability) and its utilization can be provided.

  Hereinafter, embodiments of the present invention completed based on the above knowledge will be described in detail with reference to examples. Unless otherwise stated in the embodiments and examples, J. Sambrook, EF Fritsch & T. Maniatis (Ed.), Molecular cloning, a laboratory manual (3rd edition), Cold Spring Harbor Press, Cold Spring Harbor, New York (2001); FM Ausubel, R. Brent, RE Kingston, DD Moore, JG Seidman, JA Smith, K. Struhl (Ed.), Standard Protocols in Molecular Biology, John Wiley & Sons Ltd. The method described in the protocol collection, or a modified or modified method thereof is used. In addition, when using commercially available reagent kits and measuring devices, unless otherwise explained, protocols attached to them are used.

  The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of the present specification, and those skilled in the art can easily reproduce the present invention from the description of the present specification. it can. The embodiments and specific examples of the invention described below show preferred embodiments of the present invention and are shown for illustration or explanation, and the present invention is not limited to them. It is not limited. It will be apparent to those skilled in the art that various modifications and variations can be made based on the description of the present specification within the spirit and scope of the present invention disclosed herein.

== Pharmacological action of the compound according to the invention ==
Since the compound UTKO-1 represented by the following formula (7) inhibits cell motility (migration ability), invasion ability, and proliferation, diseases caused by cell movement, diseases associated with cell invasion, It is said that it is useful as a pharmaceutical composition for diseases caused by tumor cell growth (see Japanese Patent Application Laid-Open No. 2005-330265 and International Publication No. 05/102981 pamphlet).

As described above, the present inventors represent a group represented by the following formula (m) in UTKO-1 (hereinafter referred to as “group (m)”) in the above formulas (f) to (j). The compounds (2) to (6) substituted with the group represented by formula (1) have an excellent inhibitory activity on cell motility (migration ability) and cell proliferation in the same manner as UTKO-1. It was clarified that the toxicity was strong. Therefore, in these compounds, the skeletal part (group represented by the above formula (a)) obtained by removing the group (m) from UTKO-1 is important for inhibitory activities such as cell motility, cell invasion, and cell proliferation. It became clear that it has a role.

  Therefore, the compounds (2) to (6) are a cell motility inhibitor, a cell infiltration inhibitor, and a cell proliferation inhibitor, as well as diseases caused by cell movement, diseases involving cell invasion, tumor cell proliferation. It is considered useful as a pharmaceutical composition (including preventive agents, ameliorating agents, therapeutic agents, etc.) against diseases and the like caused by.

Further, the group represented by the above formula (a) in UTKO-1 (hereinafter referred to as “group (a)”) was substituted with the groups represented by the above formulas (b) to (e). Similar to UTKO-1, compound (UTKO-2 to 5) has an action to inhibit tumor cell motility (migration ability), invasion ability, proliferation, etc., cell motility inhibitor, cell infiltration inhibition Agent, cell growth inhibitor, and pharmaceutical composition for diseases caused by cell movement, diseases involving cell invasion, diseases caused by proliferation of tumor cells, etc. 33030265 and International Publication No. 05/102981 pamphlet).

  This is because UTKO-1 is composed of two independent components, groups (a) and (m), each of which inhibits the motility (migration), invasion, and proliferation of tumor cells. In order to exhibit the function, both are necessary, but it means that the group (a) and the group (m) can be independently exchanged.

Therefore, in the above general formula (I), A is a group represented by any one of the above formulas (b) to (e), and B is represented by any one of the above formulas (f) to (j). The compound (wherein R ₁ in the formula (I) is a lower alkyl group having 1 to 4 carbon atoms) is also a tumor cell motility (migration ability), as in UTKO-1 to 5; It has an invasive ability and an action to inhibit proliferation, and is considered to be highly toxic to tumor cells. Therefore, it is effective for cell motility inhibitor, cell invasion inhibitor, cell proliferation inhibitor, and cell movement. It is considered useful as a pharmaceutical composition (including preventive agents, ameliorating agents, therapeutic agents, etc.) for diseases caused, diseases involving cell infiltration, diseases caused by proliferation of tumor cells, and the like.

Furthermore, the compound (1) in which the aldehyde group in the compound represented by the following formula (8) (Movelastin) is substituted with a hydrogen atom is superior in inhibitory activity on cell motility (migration ability) compared with Movelastin. It became clear. From this, it is considered that the compound (1) is useful as a cell motility inhibitor and a pharmaceutical composition (including a preventive agent, an improving agent, a therapeutic agent, etc.) for diseases caused by cell movement.

In addition, the compound represented by the above general formula (I) (in the formula (I), A is a group represented by any one of the above formulas (a) to (e), and B is the formula (f) To (j), and R ₁ is a lower alkyl group having 1 to 4 carbon atoms). It has the ability to inhibit tumor cell motility (migration ability), invasive ability, and proliferation, and is considered highly toxic to tumor cells. Accordingly, a compound represented by the general formula (I) (in the formula (I), A is a group represented by any one of the above formulas (a) to (e), and B is a group represented by the above formulas (f) to ( j) and R ₁ is a lower alkyl group having 1 to 4 carbon atoms.) A pharmacologically acceptable salt thereof is also a cell motility inhibitor or a cell infiltration inhibitor. , Cell growth inhibitors, and pharmaceutical compositions for diseases caused by cell movement, diseases involving cell invasion, diseases caused by tumor cell growth, etc. (including prophylactic agents, improving agents, therapeutic agents, etc.) It is considered useful.

  Examples of the diseases caused by cell movement include angiogenesis, tumor metastasis, arteriosclerosis, and diabetic retinopathy. In addition, the disease involving cell infiltration is, for example, acute myocardial infarction with inflammatory cell infiltration, tumor metastasis due to tumor cell infiltration, arteriosclerosis with leukocyte infiltration, mononuclear cell infiltration, restricted lung Diseases, gastritis, or acute interstitial pneumonia.

== Method for Producing Compound According to the Present Invention ==
The compound according to the present invention is represented, for example, by the following general formula (II) produced according to the method described in the literature (see Japanese Patent Application Laid-Open No. 2005-330265 and International Publication No. 05/102981 pamphlet). A compound, a compound represented by the general formula (IV), a compound represented by the formula (9), etc. are used as starting materials, and the reaction is carried out according to the following reaction process formula or the method described in the examples. It can manufacture by reacting according to it. In the reaction process formula, R ₁ is a lower alkyl group having 1 to 4 carbon atoms, and R ₂ is a group represented by any one of the above formulas (a) to (e).

First, an example of a method for producing a compound represented by the following general formula (III), which is a compound of the present invention, using a compound represented by the following general formula (II) will be described based on the reaction process formula 1. To do.

<Step 1: Production of compound represented by general formula (III)>
Pyridinium dichromate (PDC) is added to a methylene chloride solution of the compound represented by the general formula (II) and stirred. After stirring, the reaction solution is diluted with ethyl acetate, filtered through celite, and concentrated. The obtained concentrated solution is subjected to Florisil filtration with ethyl acetate, concentrated and dissolved in THF (tetrahydrofuran), concentrated hydrochloric acid is added and stirred. Then, saturated aqueous sodium hydrogen carbonate is added and extracted with ethyl acetate. The organic layers are combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. By purifying the obtained concentrated liquid by chromatography, a compound represented by the general formula (III) can be obtained.

Next, using a compound represented by the following general formula (IV), a method for producing a compound represented by the following general formula (V), (VI), and (VII), which is a compound of the present invention: An example will be described based on reaction process formula 2.

<Step 2: Production of compound represented by general formula (V)>
Rhodium-alumina is added to an ethyl acetate solution of the compound represented by the general formula (IV), and the mixture is stirred in hydrogen gas. After stirring, dilute with ethyl acetate, filter through celite, and concentrate. By purifying the obtained concentrated liquid by chromatography, a compound represented by the general formula (V) can be obtained.

<Step 3: Production of compound represented by general formula (VI)>
Dissolve the toluene solution of diethyl azodicarboxylate (DEAD) in THF (1 ml), add the compound represented by general formula (IV) and the THF solution of triphenylphosphine (PPh ₃ ), and stir the reaction solution. To do. A compound represented by the general formula (VI) can be obtained by purifying the obtained concentrated liquid by chromatography.

<Step 4: Production of compound represented by general formula (VII)>
After adding the compound represented by the general formula (IV), potassium carbonate, and tetrabutylammonium hydrosulfate to ethyl acetate, R ₁ I (R ₁ is a lower alkyl group having 1 to 4 carbon atoms) is added to toluene. .) Is added and stirred, and the reaction solution is concentrated. By purifying the obtained concentrated liquid by chromatography, a compound represented by the general formula (VII) can be obtained.

Next, an example of a method for producing a compound represented by the following general formula (X), which is a compound of the present invention, using the compound represented by the following formula (9) will be described based on the reaction process formula 3. .

<Step 5: Production of compound represented by formula (10)>
Ozone-oxygen gas is blown into a methylene chloride solution of 4-allyl-3,5-bis (methoxymethoxy) toluene (compound represented by formula (9)) until the color of the solution becomes light blue. Subsequently, nitrogen gas is blown to remove excess ozone with nitrogen, and then triphenylphosphine is added and stirred, and the reaction solution is concentrated under reduced pressure. A compound represented by the formula (10) can be obtained by purifying the obtained concentrated liquid by chromatography.

<Step 6: Production of compound represented by general formula (IX)>
A dimethylformamide solution of a compound represented by general formula (VIII) and a compound represented by formula (10) is dropped into a dimethylformamide (DMF) solution of chromium (II) chloride and nickel chloride (II) and stirred. . After stirring, water is added to the reaction solution and extracted with diethyl ether. The organic layers are combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. A compound represented by the general formula (IX) can be obtained by purifying the obtained concentrated liquid by chromatography.

<Step 7: Production of compound represented by general formula (X)>
Camphorsulfonic acid (CSA) is added to a methanol solution of the compound represented by the general formula (IX) and stirred. After stirring, saturated aqueous sodium hydrogen carbonate solution is added, and the mixture is extracted with ethyl acetate. The organic layers are combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. A compound represented by the general formula (X) can be obtained by purifying the obtained concentrated liquid by chromatography.

  In addition, pharmacologically acceptable salts of the compounds according to the present invention, for example, alkali metal salts (sodium salt, potassium salt, etc.), alkaline earth metal salts (magnesium salt, calcium salt, etc.), other metal salts ( Aluminum salts, etc.), inorganic salts (hydrochlorides, ammonium salts, amines, etc.), organic salts (glucosamine salts, etc.) and the like can be produced according to conventional methods.

== Drug / Pharmaceutical Composition of the Present Invention ==
Compounds represented by the above general formula (I) (in the formula (I), A is a group represented by any one of the above formulas (a) to (e), and B is a group represented by the above formulas (f) to ( j) and R ₁ is a lower alkyl group having 1 to 4 carbon atoms.) or compound (1) or a pharmacologically acceptable salt thereof as an active ingredient The composition may be administered as a pharmaceutical to humans or vertebrates other than humans, or may be used as a reagent for experiments. Such pharmaceuticals may be orally administered in the form of tablets, capsules, granules, powders, syrups, etc., or in the form of injections, suppositories, etc. by intraperitoneal or intravenous injection. It may be administered parenterally. In addition, pharmaceutical preparations are carried out by conventional methods using conventionally used formulation additives (for example, excipients, binders, lubricants, disintegrating agents, flavoring agents, solvents, stabilizers, etc.). be able to.

Hereinafter, the present invention will be specifically described by way of examples. In the following examples, nuclear magnetic resonance spectra ( ¹ H-NMR) were measured using JNM-AL300 (manufactured by JEOL). Silica gel 60N (manufactured by Kanto Chemical) was used as the silica gel for chromatography. As preparative silica gel TLC, PLC plate silica gel 60 F ₂₅₄ 0.5 mm (Merck) was used. Each reaction was performed in argon unless otherwise specified.

[Example 1]
<2,4-dihydroxy-3- {2-oxo-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] but-3-enyl} -6-methylbenzaldehyde (compound (3): UTKO- 8) Preparation>
3- {2-hydroxy-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] but-3-enyl} -2,4- (bismethoxymethoxy) -6-methylbenzaldehyde (in FIG. 1) Pyridinium dichromate (46.5 mg, 0.0618 mmol) was added to a methylene chloride solution (2 ml) of compound (11)) (22 mg, 0.0476 mmol) and stirred overnight. The reaction solution was diluted with ethyl acetate, filtered through celite, and concentrated. The obtained concentrated solution was subjected to Florisil filtration with ethyl acetate, concentrated and dissolved in THF (4 ml), concentrated hydrochloric acid (1.6 ml) was added at 0 ° C., and the mixture was stirred at room temperature for 2 hours. Thereafter, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted 3 times with ethyl acetate. The organic layers were combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting concentrated liquid was purified by preparative silica gel TLC (hexane: ethyl acetate = 3: 1), and 4.8 mg of 2,4-dihydroxy-3- {2-oxo-3- [2- (2,2 , 6-Trimethylcyclohexyl) ethyl] but-3-enyl} -6-methylbenzaldehyde (compound (3) in FIG. 1: UTKO-8).

¹ H NMR (300MHz, CDCl ₃ ): δ = 13.02 (s, 1H, Ar-OH), 10.05 (s, 1H, Ar-CHO), 9.46 (s, 1H, Ar-OH), 6.74 (s, 1H , Ar-H), 6.36 (s, 1H, -C = CH ₂ ), 6.03 (s, 1H, -C = CH ₂ ), 4.02 (s, 2H, -CH ₂ -Ar), 2.50 (s, 3H , Ar-CH ₃ ), 2.23 (tm, 1H, J = 7.5Hz, C = C-CH _2- ), 1.90 (m, 1H, Cy-H-6), 1.43 to 0.99 (m, 10H, Cy- H-1, Cy-H-3, Cy-H-4, Cy-H-5, Cy-CH _2- ), 0.92 (s, 3H, Cy-Me ₂ ), 0.86 (s, 3H, Cy-Me ₂ ), 0.82 (d, 3H, J = 6.9 Hz, Cy-Me)

[Example 2]
<Preparation of 2,4-dihydroxy-3- [2-hydroxy-3-methyl-5- (2,2,6-trimethylcyclohexyl) pentyl] -6-methylbenzaldehyde (compound (2): UTKO-7)>
2,4-Dihydroxy-3- {2-hydroxy-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] but-3-enyl} -6-methylbenzaldehyde (Compound (7): UTKO-1 ) (30 mg, 0.0801 mmol) in ethyl acetate (2 ml) was added 5% rhodium-alumina (5 mg) and stirred in hydrogen gas for 2 hours. The mixture was diluted with ethyl acetate, filtered through celite, and concentrated. The resulting concentrated liquid was purified by preparative silica gel TLC (hexane: ethyl acetate = 3: 1), and 28 mg of 2,4-dihydroxy-3- [2-hydroxy-3-methyl-5- (2, 2,6-Trimethylcyclohexyl) pentyl] -6-methylbenzaldehyde (compound (2): UTKO-7 in FIG. 2) was obtained as crystals.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 12.83, 12.82, 12.782, 12.776 (m, total 1H, Ar-OH), 10.04, 10.03 (s, total 1H, Ar-CHO), 9.42 (br.s, 1H, Ar-OH), 6.31 (s, 1H, Ar-H), 3.77 (m, 1H, -CH (OH) -), 3.07 (d, 0.5H, -CH 2 -Ar, J = 15.3Hz) 3.06 (d, 0.5H, -CH ₂ -Ar, J = 15.3Hz), 2.66 (dd, 1H, J = 8.7, 15.3 Hz, -CH ₂ -Ar), 2.50 (s, 3H, Ar-CH ₃ ) , 1.90 (m, 1H, Cy-H-6), 1.72 (s, CH ₃ -CH (OH)-), 1.72-1.01 (m, 12H, Cy-H-1, Cy-H-3, Cy- H-4, Cy-H-5, Cy-CH _2- , Cy-CH ₂ -CH _2- , CH ₃ -CH-), 0.95-0.73 (m, 12H, 4 × Me)

[Example 3]
<9-formyl-2,5-dihydro-6-hydroxy-8-methyl-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] benz [b] oxepin (compound (4): UTKO-9 Preparation of>
After 2.2 mol / l azodicarboxylate diethyltoluene solution (15 μl, 0.0330 mmol) was dissolved in THF (1 ml), UTKO-1 (compound (7)) (10 mg, 0.0267 mmol) and triphenylphosphine (8.4 mg, 0.0320 mmol) in THF (2 ml) and stirred at room temperature overnight. The reaction solution was concentrated, and the obtained concentrated solution was purified by preparative silica gel TLC (hexane: ethyl acetate = 3: 1), and 28 mg of 9-formyl-2,5-dihydro-6-hydroxy-8- Methyl-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] benz [b] oxepin (compound (4) in FIG. 3: UTKO-9) was obtained as crystals.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 10.44 (s, 1H, Ar-CHO), 6.41 (s, 1H, Ar-H), 5.69 (s, 1H, Ar-OH), 5.64 (m, 1H , -C = CH-), 4.58 (s, 2H, -CH ₂ -O-Ar), 3.44 (d, 2H, J = 5.1Hz, -CH ₂ -Ar), 2.54 (s, 3H, Ar-CH ₃ ), 1.88 (m, 1H, Cy-H-6), 1.80 (t, 2H, J = 8.4Hz, -CH ₂ -CH ₂ -C =), 1.45 to 1.04 (m, 9H, Cy-H- 1, Cy-H-3, Cy-H-4, Cy-H-5, Cy-CH _2- ), 0.93 (s, 3H, Cy-Me ₂ ), 0.88 (s, 3H, Cy-Me ₂ ) , 0.82 (d, 3H, J = 6.9Hz, Cy-Me)

[Example 4]
<2-hydroxy-3- {2-hydroxy-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] but-3-enyl} -4-methoxy-6-methylbenzaldehyde (compound (5): Preparation of UTKO-10)>
UTKO-1 (compound (7)) (20 mg, 0.0534 mmol), potassium carbonate (13.6 mg, 0.0990 mmol), and tetrabutylammonium hydrosulfate (n-Bu ₄ · HSO ₄ : 21.6 mg, 0.0639 mmol) Added to ethyl acetate (2 ml). Thereafter, 1.77 ml (0.0522 mmol) of a solution in which methyl iodide (25.1 mg, 0.177 mmol) was dissolved in toluene to make 6 ml was further added, followed by stirring at 80 ° C. overnight. After stirring, the reaction solution was concentrated to room temperature. The resulting concentrated solution was purified by preparative silica gel TLC (hexane: ethyl acetate = 2: 1) to give 2.1 mg of 2-hydroxy-3- {2-hydroxy-3- [2- (2,2,2, 6-Trimethylcyclohexyl) ethyl] but-3-enyl} -4-methoxy-6-methylbenzaldehyde (compound (5) in FIG. 4: UTKO-10) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 12.57 (s, 1H, Ar-OH), 10.13 (s, 1H, Ar-CHO), 6.32 (s, 2H, Ar-H), 5.09 (s, 1H , -C = CH ₂ ), 4.87 (s, 1H, -C = CH ₂ ), 4.28 (d, 1H, J = 9.0 Hz, -CH (OH)-), 3.90 (s, 3H, Ar-OCH ₃ ), 2.994 (d, 0.5H, J = 13.8Hz, -CH ₂ -Ar), 2.986 (d, 0.5H, J = 13.8Hz, -CH ₂ -Ar), 2.83 (dd, 0.5H, J = 9.0 , 13.8 Hz, -CH ₂ -Ar), 2.58 (s, 3H, Ar-Me), 2.18 to 2.05 (m, 2H, C = C-CH _2- ), 1.95 (m, 1H, Cy-H-6 ), 1.46 to 0.90 (m, 9H, Cy-H-1, Cy-H-3, Cy-H-4, Cy-H-5, Cy-CH _2- ), 0.90 to 0.86 (m, 9H, 3 × Me)

[Example 5]
<Preparation of [2,6-bis- (methoxymethoxy) -4-methylphenyl] acetaldehyde (compound represented by formula (10))>
To -methylene chloride solution (50 ml) of 4-allyl-3,5-bis (methoxymethoxy) toluene (compound represented by formula (9) in FIG. 5) (1.50 g, 5.31 mmol) at −78 ° C. was added ozone. -Oxygen gas was blown in until the color of the solution became light blue. Next, nitrogen gas was blown for 10 minutes to replace excess ozone with nitrogen, and triphenylphosphine (1.53 g, 5.85 mmol) was added at the same temperature, followed by stirring for 1 hour. After stirring, the temperature of the reaction solution was raised to room temperature and concentrated under reduced pressure. The obtained concentrated liquid was purified by neutral silica gel column chromatography (hexane: ethyl acetate = 10: 1), and 1.03 g of [2,6-bis- (methoxymethoxy) -4-methylphenyl] acetaldehyde (FIG. 5). A colorless oily substance was obtained from the compound represented by the formula (10).

¹ H NMR (300 MHz, CDCl ₃ ): δ = 9.63 (s, 1H, Ar-CHO), 6.65 (s, 2H, Ar-H), 5.16 (s, 4H, -OCH _2- ), 3.71 (s, 2H, Ar-CH _2- ), 3.45 (s, 6H, -OCH ₃ ), 2.34 (s, 3H, Ar-CH ₃ )

[Example 6]
<1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3- [2- (2,2,6-trimethylcyclohexyl) ethyl] -3-buten-2-ol (formula (13) Preparation of a compound represented by
At room temperature, a solution of chromium (II) chloride (337 mg, 2.74 mmol) and nickel chloride (II) (17 mg, 0.131 mmol) in dimethylformamide (5 ml) was added to trifluoromethanesulfonic acid 1- [2- (2,2,2, 6-trimethyl-cyclohexyl) ethyl] vinyl ester (compound represented by formula (12) in FIG. 5) (150 mg, 0.457 mmol) and [2,6-bis- (methoxymethoxy) -4-methylphenyl] acetaldehyde (Compound represented by formula (10) in FIG. 5) (232 mg, 0.914 mmol) in dimethylformamide (5 ml) was added dropwise and stirred overnight at the same temperature. After stirring, water was added to the reaction solution and extracted three times with diethyl ether. The organic layers were combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting concentrated liquid was purified by spherical neutral silica gel chromatography (hexane: ethyl acetate = 1: 1) to give 125 mg of 1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3- [2- (2,2,6-Trimethylcyclohexyl) ethyl] -3-buten-2-ol (a compound represented by the formula (13) in FIG. 5) was obtained as a colorless oily substance.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 6.64 (s, 2H, Ar-H), 5.19 (s, 4H, -OCH ₂ O-), 5.13 (s, 1H, -C = CH ₂ ), 4.89 (s, 1H, -C = CH ₂ ), 4.27 (d, 1H, J = 9.1 Hz, -CH-OH), 3.48 (s, 6H, -OCH ₃ ), 3.05 (d, 1H, J = 13.8 Hz , -CH ₂ -Ar), 2.85 (dd, 1H, J = 9.1, 13.8Hz, -CH ₂ -Ar), 2.54 (br.s, 1H, -OH), 2.31 (s, 3H, Ar-Me) , 2.11 (m, 2H, C = C-CH _2- ), 1.95 (m, 1H, Cy-H-6), 1.48 to 1.06 (m, 9H, Cy-H-1, Cy-H-3, Cy -H-4, Cy-H-5, Cy-CH _2- ), 0.97 to 0.88 (m, 9H, 3 × Me)

[Example 7]
<2- {2-hydroxy-3- [2- (2,2,6-trimethylcyclohexyl) ethyl] but-3-ene} -5-methyl-1,3-benzenediol (compound (6): UTKO- 12) Preparation>
1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3- [2- (2,2,6-trimethylcyclohexyl) ethyl] -3-buten-2-ol (formula in FIG. 5) To a methanol solution (3 ml) of the compound represented by (13) (55 mg, 0.127 mmol), camphorsulfonic acid (17 mg, 0.0732 mmol) was added and stirred overnight. Saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted 3 times with ethyl acetate. The organic layers were combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting concentrated liquid was purified by preparative silica gel TLC (hexane: ethyl acetate = 2: 1) to give 22 mg of 2- {2-hydroxy-3- [2- (2,2,6-trimethylcyclohexyl). Ethyl] but-3-ene} -5-methyl-1,3-benzenediol (compound (6) in FIG. 5: UTKO-12) was obtained as white crystals.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 6.31 (s, 2H, Ar-OH), 6.28 (s, 2H, Ar-H), 5.09 (s, 1H, -C = CH ₂ ), 4.91 (s , 1H, -C = CH ₂ ), 4.37 (d, 1H, J = 8.7 Hz, -CH-OH), 3.11 (d, 1H, J = 14.7 Hz, -CH ₂ -Ar), 2.72 (dd, 0.5 H, J = 8.7Hz, 14.7 Hz, -CH ₂ -Ar), 2.71 (dd, 0.5H, J = 8.7, 14.7 Hz, -CH ₂ -Ar), 2.22 (s, 3H, Ar-CH ₃ ), 2.15 to 2.05 (m, 2H, C = C-CH _2- ), 1.93 (m, 1H, Cy-H-6), 1.48 to 1.01 (m, 9H Cy-H-1, Cy-H-3, Cy -H-4, Cy-H-5, Cy-CH _2- ), 0.96 (s, 3H, Cy-Me ₂ ), 0.90 (s, 3H, Cy-Me ₂ ), 0.87 (d, 3H, J = (6.9 Hz, Cy-Me)

[Example 8]
<1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3-[(6R, 7S, 8R) -6,7,8-trimethyl-1,4-dioxaspiro [4.5] dec-7 Preparation of -yl] -3-buten-2-ol (compound represented by formula (15))>
At room temperature, a solution of chromium (II) chloride (286 mg, 2.33 mmol) and nickel chloride (II) (14 mg, 0.108 mmol) in dimethylformamide (20 ml) was added to trifluoromethanesulfonic acid (6R, 7S, 8R) -1- [2- (6,7,8-trimethyl-1,4-dioxaspiro [4.5] dec-7-yl) ethyl] vinyl ester (compound represented by formula (14) in FIG. 6) (150 mg, 0.388 mmol ) And [2,6-bis- (methoxymethoxy) -4-methylphenyl] acetaldehyde (compound represented by formula (10)) (197 mg, 0.776 mmol) in dimethylformamide (20 ml) were added dropwise, and the same temperature And stirred overnight. After stirring, water was added to the reaction solution and extracted three times with diethyl ether. The organic layers were combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting concentrated liquid was purified by neutral silica gel column chromatography (hexane: ethyl acetate = 1: 1), and 110 mg of 1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3- [(6R, 7S, 8R) -6,7,8-trimethyl-1,4-dioxaspiro [4.5] dec-7-yl] -3-buten-2-ol (represented by the formula (15) in FIG. 6) Compound) was obtained as a colorless oil.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 6.65 (s, 2H, Ar-H), 5.20 (s, 4H, -OCH ₂ O-), 5.11 (s, 1H, -C = CH ₂ ), 4.86 (s, 1H, -C = CH ₂ ), 4.26 (m, 1H, -CH-OH), 4.04 to 3.77 (m, 4H, -O-CH ₂ -CH ₂ -O-), 3.48 (s, 6H , -OCH ₃ ), 3.03 (d, 1H, J = 13.5 Hz, -CH ₂ -Ar), 2.83 (dd, 1H, J = 9.6, 13.5 Hz, -CH ₂ -Ar), 2.55 (dd, 1H, J = 5.2, 6.9 Hz, -CH (Me) -CO-), 2.32 (s, 3H, Ar-Me), 2.12 to 1.75 (m, 4H, -CH ₂ -CH ₂ -CH (Me)-, C = C-CH _2- ), 1.54-1.26 (m, 5H, -CH ₂ -CH (Me)-, -CH ₂ -CH (Me)-, C = C-CH ₂ -CH _2- ), 0.91 ~ 0.79 (m, 9H, 3 × Me)

[Example 9]
<(2R, 3S, 4R) -3- [4-hydroxy-5- (2,6-dihydroxy-4-methylphenyl) -3-methylenepentyl] -2,3,4-trimethylcyclohexanone (compound (1) : Preparation of UTKO-11)>
1- [2,6-bis (methoxymethoxy) -4-methylphenyl] -3-[(6R, 7S, 8R) -6,7,8-trimethyl-1,4-dioxaspiro [4.5] dec-7- Ilf] -3-buten-2-ol (compound represented by formula (15) in FIG. 6) (20 mg, 0.0406 mmol) in methanol solution (4 ml) was added camphorsulfonic acid (20 mg, 0.0861 mmol). And stirred for 8 hours. Saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted 3 times with ethyl acetate. The organic layers were combined, washed with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained concentrated liquid was purified by preparative silica gel TLC (hexane: ethyl acetate = 2: 1), and 1.4 mg of (2R, 3S, 4R) -3- [4-hydroxy-5- (2,6- Dihydroxy-4-methylphenyl) -3-methylenepentyl] -2,3,4-trimethylcyclohexanone (compound (1): UTKO-11) was obtained as colorless crystals.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 6.29 (s, 4H, Ar-OH, Ar-H), 5.11 (d, 1H, J = 3.0 Hz, -C = CH ₂ ), 4.91 (s, 1H , -C = CH ₂ ), 4.40 (d, 1H, J = 8.1 Hz, -CH-OH), 3.12 (d, 1H, J = 14.4 Hz, -CH ₂ -Ar), 2.80 to 2.72 (m, 1H , -CH ₂ -Ar), 2.55 to 1.02 (m, 10H, protons of cyclohexanone, C = C-CH _2- , C = C-CH ₂ -CH _2- ), 0.95 to 0.86 (m, 6H, 2- Me, 4-Me), 0.61 (s, 3H, 3-Me)

[Example 10]
In this example, the effects of UTKO-7 to 12 obtained in Examples 1, 2, 3, 4, 7, and 9 on cell migration ability (motility), cell proliferation, farnesyltransferase, etc. In addition, the following experiment was conducted to examine the cytotoxicity of UTKO-7-12.

<Inhibitory effect on cell migration ability>
In order to investigate whether UTKO-7-12 has an inhibitory effect on the migration ability of tumor cells in the same manner as UTKO-1-5 described above, migration ability was measured by wound healing assay using UTKO-7-12. Was measured.

To a 48-well plate, 500 μl of a suspension of EC17 cells derived from human esophageal cancer (1.5 × 10 ⁵ cells / ml; RPMI1640 medium) was added and cultured at 37 ° C. for 24 hours. Thereafter, the center of the cell surface of each well was scratched with a micropipette tip, and the culture supernatant was immediately removed, and PBS ⁻ (Ca ²⁺ , Mg ²⁺ -free PBS; 8 g / l NaCl, 0.2 Wash with 1 μg of serum (FBS; Tissue Culture Biologicals) carefully taking care not to remove cells with 300 μl of g / l KCl, 0.916 g / l Na ₂ HPO ₄ , 0.2 g / l KH ₂ PO _4. 500 μl of RPMI1640 medium containing Then, add the above UTKO-1 ~ 5 or UTKO-7 ~ 12, incubate at 37 ° C for 24 hours, and then observe under a microscope to confirm how much the wound damaged by the micropipette tip is buried. Evaluated the ability to migrate.

<Cell toxicity test>
The above-mentioned UTKO-1 to 5 are known to be highly toxic to tumor cells as compared to movelastin. Therefore, in order to examine whether UTKO-7-12 is highly toxic to cells, the cytotoxicity of UTKO-7-12 was examined by trypan blue extracellular excretion test method. To a 96-well plate, 200 μl of EC17 cell suspension (2.5 × 10 ⁴ cells / ml; RPMI1640 medium) was added and cultured at 37 ° C. for 24 hours, and the above UTKO-1-5 or UTKO-7-12 Was added and cultured at 37 ° C. for 3 days. After culturing, replace with 50 μl trypsin solution, peel off the cells, dissociate the cells by pipetting, add 5 times concentrated trypan blue solution (9 g / l NaCl, 4 g / l trypan blue), mix well, Cells stained with a counting plate (Elma) were counted. The cell survival rate was determined by the following formula.
(Cell viability) = ((viable cell count) / (total cell count)) × 100

<Inhibitory effect on cell proliferation>
To examine the inhibitory effect of UTKO-7-12 on cell proliferation, MTT assay was performed. To a 96-well plate, 200 μl of EC17 cell suspension (2.5 × 10 ⁴ cells / ml; RPMI1640 medium) was added and cultured at 37 ° C. for 24 hours, and the above UTKO-1-5 or UTKO-7-12 Was added and cultured at 37 ° C. for 3 days. Thereafter, 10 μl of MTT (3- (4,5-dimethlthiazol-2-yl) -2,5-diphenyltetrazolium bromide; Sigma) solution (adjusted to 5 mg / ml with PBS ⁻ ) was added at 4 ° C. at 37 ° C. Incubate for hours. After incubation, remove the medium, dissolve formazan precipitate (dark blue insoluble substance in which yellow water-soluble MTT is reduced by succinate dehydrogenase present in the mitochondria of cells) with 100 μl dimethyl sulfoxide, and microplate The absorbance (measurement wavelength 570 nm, reference side 620 nm) was measured using a reader (Tosoh Corporation) to evaluate the growth inhibitory action.

<Inhibitory effect on farnesyltransferase>
In order to examine the inhibitory effect of UTKO-7-12 on farnesyltransferase, FTase assay was performed. As FTase, EC17 cells were roughly purified as follows.

EC17 cells lysed with Lysis buffer (10 mM Hepes pH 7.4, 1 mM MgCl ₂ , 1 mM EGTA, 1 mM PMSF, 1 μM leupeptin, 10 mM sodium pyrophosphate, 0.1 mM sodium orthovanadate (V), and 100 mM NaF) on ice After standing for 30 minutes, the mixture was centrifuged at 10,000 g for 15 minutes. Thereafter, the supernatant was collected and further centrifuged at 100,000 g for 45 minutes, and then 60% saturated ammonium sulfate was added to the supernatant, and the precipitated fraction was centrifuged at 10,000 g for 30 minutes. The resulting precipitate was collected, dissolved in dialysis buffer (50 mM Tris-HCl pH 7.5, 1 mM DTT, and 0.02 mM MgCl ₂ ), dialyzed against dialysis buffer for 12 hours, and stored at −80 ° C.

  Next, H-ras gene was amplified using RT-PCR method using mRNA extracted from EC17 cells and subcloned into pGEX-2T vector (TaKaRa). After introducing the subcloned plasmid into E. coli DH5-α strain, the transformed E. coli was cultured, and expression was induced with IPTG (isopropyl-1-thio-β-D-galactopyranoside; STRATEGENE). The expressed GST-H-Ras protein was then purified using glutathione-agarose beads (Sigma).

As described above, 30 μl (30 μg) of FTase roughly purified from EC17 cells and 10 μl (10 μg) of GST-H-Ras protein were added to 10 μl of reaction buffer (50 mM Tris-HCl pH 7.5, 1 mM MgCl ₂ , and 4 mM DTT). Double concentrated solution) 6 μl, [ ³ H] -FPP (final concentration 0.06 μM (596 GBq / mmol); New England Nuclear) 0.2 μl, and drug (each UTKO-1-5 or UTKO-7-12 above) ) 3 μl and 1.7 ml Eppendorf tube were mixed and incubated at 37 ° C. for 1 hour. After incubation, the reaction was stopped by adding 0.5 ml of methanol containing 1% SDS (sodium dodecyl sulfate) and 0.5 ml of 30% TCA, and left on ice for 1 hour, using a cell harvester (ADVANTEC). The acid insoluble fraction was trapped with a glass fiber filter (Whatman GF / C filter) and washed with 6% TCA. Thereafter, the filter was dried, and the radioactivity contained in the acid-insoluble fraction was measured with a liquid scintillation counter (Beckman Coulter) to calculate the FTase activity. The calculation of FTase activity was performed by subtracting the blank value from all the values using FTase whose enzyme activity was deactivated by heat treatment at 100 ° C. for 10 minutes.

The results of the above experiment are shown in Table 1. In Table 1, “a” represents the concentration at which migration ability was completely inhibited, and “b” represents the IC ₅₀ value.

表１に示すように、UTKO-7〜12は、UTKO-1〜5と同様に、FTaseに対する阻害活性を示さないものの、腫瘍細胞の遊走能に対してUTKO-1〜5より優れた阻害活性を有することがわかった。また、UTKO-7〜10及びUTKO-12は、腫瘍細胞の増殖に対してUTKO-1〜5より優れた阻害活性を有し、腫瘍細胞に対する毒性がUTKO-1〜5より高いことが明らかになった。このことから、UTKO-7〜10及びUTKO-12は薬剤として特に有用であることが示された。なお、薬剤を含まないネガティブコントロールは阻害活性が検出されなかった。 [Table 1]

As shown in Table 1, UTKO-7-12, like UTKO-1-5, show no inhibitory activity against FTase, but superior inhibitory activity to UTKO-1-5 over tumor cell migration ability. It was found to have In addition, UTKO-7-10 and UTKO-12 have superior inhibitory activity against tumor cell growth than UTKO-1-5, and are clearly more toxic to tumor cells than UTKO-1-5 became. This indicates that UTKO-7 to 10 and UTKO-12 are particularly useful as drugs. In addition, no inhibitory activity was detected in the negative control containing no drug.

In one Example of this invention, it is a figure which shows the manufacture process of a compound (3). In one Example of this invention, it is a figure which shows the manufacture process of a compound (2). In one Example of this invention, it is a figure which shows the manufacture process of a compound (4). In one Example of this invention, it is a figure which shows the manufacture process of a compound (5). In one Example of this invention, it is a figure which shows the manufacture process of a compound (6). In one Example of this invention, it is a figure which shows the manufacture process of a compound (1).

Claims (13)

A compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof.

(In the formula (I), A is a group represented by any of the following formulas (a) to (e), and B is a group represented by any of the following formulas (f) to (j): R ₁ is a lower alkyl group having 1 to 4 carbon atoms.)

A compound represented by the following formula (1) or a pharmacologically acceptable salt thereof:

Any compound represented by the following formulas (2) to (6) or a pharmacologically acceptable salt thereof.

A pharmaceutical composition comprising as an active ingredient a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof.

(In the formula (I), A is a group represented by any of the following formulas (a) to (e), and B is a group represented by any of the following formulas (f) to (j): R ₁ is a lower alkyl group having 1 to 4 carbon atoms.)

A pharmaceutical composition comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

A pharmaceutical composition comprising any compound represented by the following formulas (2) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

A cell motility inhibitor comprising a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

(In the formula (I), A is a group represented by any of the following formulas (a) to (e), and B is a group represented by any of the following formulas (f) to (j): R ₁ is a lower alkyl group having 1 to 4 carbon atoms.)

A cell motility inhibitor comprising a compound represented by the following formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

A cell motility inhibitor comprising as an active ingredient any compound represented by the following formulas (2) to (6) or a pharmacologically acceptable salt thereof:

A drug that inhibits cell invasion containing a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient.

(In the formula (I), A is a group represented by any of the following formulas (a) to (e), and B is a group represented by any of the following formulas (f) to (j): R ₁ is a lower alkyl group having 1 to 4 carbon atoms.)

A drug that inhibits cell invasion containing any compound represented by the following formulas (2) to (6) or a pharmacologically acceptable salt thereof as an active ingredient.

A cell growth inhibitor comprising as an active ingredient a compound represented by the following general formula (I) or a pharmacologically acceptable salt thereof:

(In the formula (I), A is a group represented by any of the following formulas (a) to (e), and B is a group represented by any of the following formulas (f) to (j): R ₁ is a lower alkyl group having 1 to 4 carbon atoms.)

A cell growth inhibitor comprising as an active ingredient any compound represented by the following formulas (2) to (6) or a pharmacologically acceptable salt thereof:

Images