JP2005306865A - Cis- and trans-stilbene compounds each containing boric acid group and their preparation methods - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a compound which has a basic skeleton of combretastatin, a basic skeleton of Resveratrol or a skeleton similar to the basic skeleton of Resveratrol, whose physiological activity hardly deteriorates or whose in vivo behavior can be easily analyzed, and to provide a preparation method for the compound. <P>SOLUTION: The compound is a cis-stilbene compound containing a boric acid group, which is represented by formula (1) (wherein R1 is hydrogen atom or methoxy group; R2 is hydrogen atom, a boric acid group or methoxy group; R3 is hydrogen atom, a boric acid group or methoxy group, provided that R1, R2 and R3 are identical to or different from each other and either R2 or R3 is a boric acid). <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  TECHNICAL FIELD The present invention relates to a boric acid group-containing cis-stilbene compound and a method for producing the same, and a boric acid group-containing trans-stilbene compound and a method for producing the same. Boric acid group-containing cis-stilbene compound having skeleton and method for producing the same, and boric acid having a basic skeleton of resveratrol having various antitumor activities such as apoptosis-inducing action or a skeleton similar to the basic skeleton of resveratrol The present invention relates to a group-containing trans-stilbene compound and a method for producing the same.

  Tubulin polymerization inhibitors have the effect of inhibiting cancer cell cell division and killing cancer cells by damaging vascular endothelial cells and blocking blood flow. Has attracted attention as a new target substance.

  Currently, as a compound having an inhibitory effect on tubulin polymerization, combretastatin A-4 (the following formula (16)):

And combretastatin A-2 (the following formula (17)):

(Hereinafter, both compounds are collectively referred to as combretastatin compounds). However, the combretastatin compound is poor in water solubility and cannot be administered as a drug as it is. Therefore, conventionally, in order to use the combretastatin compound as a pharmaceutical, it has been performed to impart water solubility to the combretastatin compound.

  Non-Patent Document 1 discloses a combretastatin A-4 prodrug in which the hydroxyl group of combretastatin A-4 is converted to a sodium phosphate salt as a compound that imparts water solubility to the combretastatin compound. As of 2002, the prodrug has progressed to clinical trial Phase II.

Non-Patent Document 2 discloses a prodrug obtained by converting the hydroxyl group of combretastatin A-2 to amino hydrochloride.
Pettit, GR et.al, J. Med. Chem., 2003, 46, 525. Pettit, GR et.al, Anti-Cancer Drug Design, 2001, 16, 185.

  In addition, resveratrol (Resevertrol, (E) -3,4 ', 5-trihydroxystilbene, the following formula (18)), which is contained in a lot of grape plant skins;

Is a trans-stilbene compound that has recently attracted attention as a compound that has been recognized for various antitumor activities such as apoptosis-inducing action that promotes the death of tumor cells and prevents proliferation.

  A conventional drug exhibits physiological activity by forming a hydrogen bond with an amino acid, a hydroxyl group, a thiol group, or the like present in a target receptor protein. However, since the hydrogen bond is reversible, the drug is likely to be released from the target receptor protein, so that the conventional drug has a problem that the physiological activity is likely to be lowered.

  In addition, analysis of drug behavior in vivo is very important in order to clarify problems such as side effects and safety. However, most of the drugs currently being developed are composed of elements existing in the living body, that is, elements such as hydrogen, carbon, nitrogen, oxygen, sulfur, and phosphorus. It was not easy. For example, when the prodrug disclosed in Non-Patent Document 1 or 2 is administered with a drug, it is difficult to analyze the behavior in vivo.

  Accordingly, an object of the present invention is to provide a compound having a basic skeleton of combretastatin, which is difficult to reduce physiological activity, or a compound whose behavior in vivo can be easily analyzed, and a method for producing the compound Is to provide. Another object of the present invention is a compound having a basic skeleton of resveratrol or a skeleton similar to the basic skeleton of resveratrol, a compound in which physiological activity is difficult to decrease, or a compound whose behavior in vivo is easy to analyze And a method for producing the compound.

  As a result of intensive studies to solve the above-described problems in the prior art, the present inventors have found that a boric acid group-containing cis-stilbene compound in which a boric acid group is introduced into the basic skeleton of combretastatin, or resveratrol The boric acid group-containing trans-stilbene compound in which a boric acid group is introduced into a basic skeleton of the above or a basic skeleton of resveratrol has a solubility in water as compared with (1) a combretastatin compound or resveratrol. (2) Since the boron atom can be traced by cross-sectional imaging by α-autoradiograph or the like, the boric acid group-containing cis-stilbene compound or the boric acid group-containing trans-stilbene compound The inventors have found that behavioral analysis in a living body can be easily performed, and have completed the present invention.

  That is, the present invention (1) includes the following general formula (1):

(In the formula, R ¹ represents a hydrogen atom or a methoxy group, R ² represents a hydrogen atom, a boric acid group or a methoxy group, R ³ represents a hydrogen atom, a boric acid group or a methoxy group, and R ¹ , R ² And R ³ may be the same or different, and either R ² or R ³ is a boric acid group.) Provides a boric acid group-containing cis-stilbene compound.

  Further, the present invention (2) includes the following general formula (2):

(Wherein R ¹ represents a hydrogen atom or a methoxy group) and the following general formula (3):

(Wherein X represents a leaving group, R ⁵ represents a hydrogen atom, a protected boric acid group or a methoxy group, R ⁶ represents a hydrogen atom, a protected boric acid group or a methoxy group, and R ⁵ or R ⁶ Any one is a protected boric acid group.) A coupling step (A) for obtaining a protected boric acid group-containing disubstituted alkyne compound by reacting a protected boric acid group-containing benzene compound represented by Hydrogenation of the protected boric acid group-containing disubstituted alkyne compound obtained in A) to obtain a protected boric acid group-containing cis-stilbene compound, and the protected boric acid group-containing cis obtained in the hydrogenation step -The manufacturing method of the boric acid group containing cis-stilbene compound which has the deprotection process which remove | eliminates the protective group of the boric acid group of a stilbene compound, and obtains a boric acid group containing cis-stilbene compound is provided.

  Further, the present invention (3) includes the following general formula (4):

(Wherein R ⁷ represents a methoxy group, and n is 2 or 3). This provides a boric acid group-containing trans-stilbene compound (A) represented by:

  Further, the present invention (4) includes the following general formula (5):

(Wherein R ⁸ represents a hydroxyl group, and m is 2 or 3). A boric acid group-containing trans-stilbene compound (B) represented by formula (B) is provided.

  Further, the present invention (5) includes the following general formula (6):

(Wherein R ⁷ represents a methoxy group, Y represents a halogenated phosphonium group, a phosphoric acid group or a phosphoric acid ester group, and n is 2 or 3) and the following general formula ( 7):

(Wherein Z represents a leaving group). A coupling step (B) for obtaining a leaving group-containing trans-stilbene compound by reacting with a benzaldehyde derivative represented by the following formula: obtained in the coupling step (B). A boric acid group-containing trans-stilbene compound having a boric acid group introduction step in which the leaving group of the leaving group-containing trans-stilbene compound is substituted with a boric acid group to obtain a boric acid group-containing trans-stilbene compound (A) The manufacturing method of (A) is provided.

  Further, the present invention (6) includes the following general formula (6):

(Wherein R ⁷ represents a methoxy group, Y represents a halogenated phosphonium group, a phosphoric acid group or a phosphoric acid ester group, and n is 2 or 3) and the following general formula ( 7):

(Wherein Z represents a leaving group). A coupling step (B) for obtaining a leaving group-containing trans-stilbene compound by reacting with a benzaldehyde derivative represented by the following formula: obtained in the coupling step (B). The leaving group of the leaving group-containing trans-stilbene compound is substituted with a boric acid group to obtain a boric acid group-containing trans-stilbene compound (A). A boric acid group-containing trans-stilbene having a hydroxylation step in which a methoxy group of the boric acid group-containing trans-stilbene compound (A) is converted into a hydroxyl group to obtain a boric acid group-containing trans-stilbene compound (B) The manufacturing method of a compound (B) is provided.

  The boric acid group-containing cis-stilbene compound of the present invention (hereinafter also referred to as a boric acid group-containing cis-stilbene compound (1)) has a basic skeleton of combretastatin and is unlikely to have decreased physiological activity, or Behavioral analysis in vivo is easy. Moreover, the manufacturing method of the boric acid group containing stilbene compound of this invention is used suitably for manufacturing this boric acid group containing cis-stilbene compound (1). Further, the boric acid group-containing trans-stilbene compound (A) of the present invention (hereinafter also simply referred to as a boric acid group-containing trans-stilbene compound (A)) and the boric acid group-containing trans-stilbene compound of the present invention (B ) (Hereinafter also referred to simply as a boric acid group-containing trans-stilbene compound (B)) has a basic skeleton of resveratrol or a skeleton similar to the basic skeleton of resveratrol, and its physiological activity is unlikely to decrease. Or, behavioral analysis in a living body is easy. Moreover, the manufacturing method of the boric acid group containing trans-stilbene compound (A) of this invention is used suitably for manufacturing this boric acid group containing trans-stilbene compound (A), and the boric acid of this invention is used. The method for producing the group-containing trans-stilbene compound (B) is suitably used for producing the boric acid group-containing trans-stilbene compound (B).

As shown in the general formula (1), the boric acid group-containing cis-stilbene compound (1) according to the present invention has a basic skeleton of combretastatin (in the general formula (1), R ¹ , R ² and R ³ having a portion) excluding and to either the 3'-position or 4'-position of a boric acid group. The boric acid group-containing cis-stilbene compound (1) may have a methoxy group at the 4-position, 3′-position or 4′-position. Specific examples of the boric acid group-containing cis-stilbene compound (1) include 3,4,4 ′, 5-tetramethoxy-3 ′-(dihydroxylboryl) cis-stilbene, 3,3 ′, 4,5 -Tetramethoxy-4 '-(dihydroxylboryl) cis-stilbene, 3,4,5-trimethoxy-4'-(dihydroxylboryl) cis-stilbene, 3,4,5-trimethoxy-3 '-(dihydroxyl Boryl) cis-stilbene, 3,3 ′, 5-trimethoxy-4 ′-(dihydroxylboryl) cis-stilbene, 3,4 ′, 5-trimethoxy-3 ′-(dihydroxylboryl) cis-stilbene, 3, 5-Dimethoxy-4 '-(dihydroxylboryl) cis-stilbene and 3,5-dimethoxy-3'-(dihydroxylboryl) cis-stilbene. Since the boric acid group-containing cis-stilbene compound (1) has the basic skeleton of the combretastatin A-4, it has tubulin polymerization inhibitory activity.

  The boric acid group-containing cis-stilbene compound (1) is a cis form in which two phenyl groups are on the same side with respect to a double bond in the molecule, but contains a trans form in which the phenyl group is on the opposite side. You may do. The content ratio of the trans isomer relative to the cis isomer is preferably 0.1 to 10% from the viewpoint of few side effects when administered as a drug, and particularly preferably 0.1 to 1%.

  Further, the purity of the boric acid group-containing cis-stilbene compound (1) is preferably 0.1 to 10% from the viewpoint of few side effects when administered as a drug, particularly preferably 0.1 to 1%. It is.

As shown in the general formula (4), the boric acid group-containing trans-stilbene compound (A) according to the present invention has a skeleton similar to the basic skeleton of resveratrol (in the general formula (4), R ⁷ , And a portion excluding a boric acid group), a boric acid group at the 3′-position, and a benzene ring different from the benzene ring having the boric acid group has 2 or 3 methoxy groups . That is, the boric acid group-containing trans-stilbene compound (A) has a methoxy group at two or three positions among the 2-position, 3-position, 4-position, 5-position, and 6-position. It has a boric acid group at the 3′-position and a methoxy group at the 4′-position. Specific examples of the boric acid group-containing trans-stilbene compound (A) include 2,3,4'-trimethoxy-3 '-(dihydroxylboryl) trans-stilbene, 2,4,4'-trimethoxy-3'. -(Dihydroxylboryl) trans-stilbene, 2,4 ', 5-trimethoxy-3'-(dihydroxylboryl) trans-stilbene, 2,4 ', 6-trimethoxy-3'-(dihydroxylboryl) trans- Stilbene, 3,4,4′-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 3,4 ′, 5-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 3,4 ′, 6 -Trimethoxy-3 '-(dihydroxylboryl) trans-stilbene, 4,4', 5-trimethoxy-3 '-(dihydroxylboryl) tolane Stilbene, 4,4 ′, 6-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 4 ′, 5,6-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 2,3 4,4′-tetramethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 2,3,4 ′, 5-tetramethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 2,3,4 ′ , 6-Tetramethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 3,4,4 ′, 5-tetramethoxy-3 ′-(dihydroxylboryl) trans-stilbene, 3,4,4 ′, 6 -Tetramethoxy-3 '-(dihydroxylboryl) trans-stilbene, 4,4', 5,6-tetramethoxy-3 '-(dihydroxylboryl) trans-stilbene Can be mentioned. Since the boric acid group-containing trans-stilbene compound (A) has a skeleton similar to the basic skeleton of resveratrol, it has antitumor activity.

  The boric acid group-containing trans-stilbene compound (A) is a trans isomer, but may also contain a cis isomer. The content ratio of the cis isomer to the trans isomer is preferably 0.1 to 10% in terms of few side effects when administered as a drug, and particularly preferably 0.1 to 1.0%.

  Further, the purity of the boric acid group-containing trans-stilbene compound (A) is preferably 0.1 to 10% from the viewpoint of few side effects when administered as a drug, and particularly preferably 0.1 to 1.%. 0%.

As shown in the general formula (5), the boric acid group-containing trans-stilbene compound (B) according to the present invention has a basic skeleton of resveratrol (in the general formula (5), R ⁸ and a boric acid group). And a boric acid group at the 3′-position, and a benzene ring different from the benzene ring having the boric acid group has 2 or 3 hydroxyl groups. That is, the boric acid group-containing trans-stilbene compound (B) has a hydroxyl group at two or three positions among the 2-position, 3-position, 4-position, 5-position, and 6-position. It has a boric acid group at the 3′-position and a hydroxyl group at the 4′-position. Specific examples of the boric acid group-containing trans-stilbene compound (B) include 2,3,4'-trihydroxyl-3 '-(dihydroxylboryl) trans-stilbene, 2,4,4'-trihydroxyl- 3 ′-(dihydroxylboryl) trans-stilbene, 2,4 ′, 5-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 2,4 ′, 6-trihydroxyl-3 ′-(dihydroxyl Boryl) trans-stilbene, 3,4,4′-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 3,4 ′, 5-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 3,4 ′, 6-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 4,4 ′, 5-trihydroxyl-3 ′ -(Dihydroxylboryl) trans-stilbene, 4,4 ', 6-trihydroxyl-3'-(dihydroxylboryl) trans-stilbene, 4 ', 5,6-trihydroxyl-3'-(dihydroxylboryl) Trans-stilbene, 2,3,4,4′-tetrahydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 2,3,4 ′, 5-tetrahydroxyl-3 ′-(dihydroxylboryl) trans- Stilbene, 2,3,4 ', 6-tetrahydroxyl-3'-(dihydroxylboryl) trans-stilbene, 3,4,4 ', 5-tetrahydroxyl-3'-(dihydroxylboryl) trans-stilbene, 3,4,4 ′, 6-tetrahydroxyl-3 ′-(dihydroxylboryl) trans-stilbene, 4,4 ′, 5,6-te La hydroxyl -3 '- (dihydroxyl boryl) trans - include stilbene. Since the boric acid group-containing trans-stilbene compound (B) has the basic skeleton of resveratrol, it has antitumor activity.

  The boric acid group-containing trans-stilbene compound (B) is a trans form, but may contain a cis form. The content ratio of the cis isomer to the trans isomer is preferably 0.1 to 10% in terms of few side effects when administered as a drug, and particularly preferably 0.1 to 1.0%.

  Further, the purity of the boric acid group-containing trans-stilbene compound (B) is preferably 0.1 to 10% from the viewpoint of few side effects when administered as a drug, and particularly preferably 0.1 to 1.%. 0%.

  The boric acid group-containing cis-stilbene compound (1) has a higher water solubility than the combretastatin A-4 because either the 3'-position or the 4'-position is a boric acid group. Further, the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) have a higher water solubility than the resveratrol because the 3'-position is a boric acid group. . Furthermore, the boric acid group reacts with a basic compound such as sodium hydroxide, is easily converted into a highly water-soluble salt, and reacts with a polyhydric alcohol compound to easily form a complex. Can do. Therefore, the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) can easily improve water solubility. It is easy to make a medicinal product by forming a complex with a saccharide or the like.

  Moreover, the boron atom in the boric acid group of the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) is: Since it has a vacant orbit without electrons, the vacant orbit receives an unshared electron pair such as a nitrogen atom of the amino group of the target receptor protein, an oxygen atom of the hydroxyl group, or a sulfur atom of the thiol group, A covalent bond can be formed. As shown in the following reaction formula (8), a compound having a boric acid group reacts with a primary amine compound, a secondary amine compound, or a compound having a hydroxyl group, and by a covalent bond, the primary amine compound, It binds to the secondary amine compound or the compound having a hydroxyl group.

  Since the covalent bond is an irreversible bond, the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A), and the boric acid group-containing trans-stilbene Compound (B) can bind strongly to the target receptor protein as compared with a conventional drug bound by a reversible hydrogen bond. Therefore, the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) are covalently bonded to the target receptor protein. It is considered that it is a compound capable of inducing a new physiological activity that the effect as a drug is hardly lowered by forming.

  Further, since the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) contain a boron atom. After administration into a living body, the boric acid group-containing cis-stilbene compound (1), boric acid can be obtained by, for example, imaging with α-autoradiograph or measuring the boron concentration in each tissue using ICP. The group-containing trans-stilbene compound (A) or the boric acid group-containing trans-stilbene compound (B) can be traced. Therefore, the boric acid group-containing cis-stilbene compound (1), the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) can be easily analyzed in vivo. is there.

  In addition, since boric acid groups are easily converted into other functional groups, the boric acid group-containing cis-stilbene compound (1) is suitable for production of derivatives having the basic skeleton of combretastatin A-4. In addition, the boric acid group-containing trans-stilbene compound (A) and the boric acid group-containing trans-stilbene compound (B) are a skeleton similar to the basic skeleton of resveratrol or the basic skeleton of resveratrol. It is suitably used for the production of derivatives having Examples of the functional group conversion method include Suzuki coupling reaction. The Suzuki coupling reaction is a reaction for producing a compound having a target functional group introduced by reacting a boric acid group-containing compound with a halogenated organic compound as shown in the following reaction formula (9). .

(In the formula, R and R ″ represent an aryl group, an alkenyl group or an alkyl group, and R ′ represents a hydrogen atom or an alkyl group.)

  Next, the manufacturing method of the boric acid group containing cis-stilbene compound of this invention is demonstrated with reference to following Reaction formula (12).

  The method for producing a boric acid group-containing cis-stilbene compound of the present invention includes a monosubstituted alkyne compound represented by the general formula (2) (hereinafter also referred to as a monosubstituted alkyne compound (2)) and the general formula (3). A coupling step (A) in which a protected boric acid group-containing benzene compound (hereinafter also referred to as a protected boric acid group-containing benzene compound (3)) is reacted to give a protected boric acid group-containing disubstituted alkyne compound (10) (A) ), Hydrogenating the protected boric acid group-containing disubstituted alkyne compound (10) obtained in the coupling step (A) to obtain a protected boric acid group-containing cis-stilbene compound (11), and A boric acid group-containing cis-stilbene compound (1) is obtained by deprotecting the protective group of the boric acid group of the protected boric acid group-containing cis-stilbene compound (11) obtained in the hydrogenation step. With a deprotection step that.

  The reaction of the monosubstituted alkyne compound (2) and the protected boric acid group-containing benzene compound (3) in the coupling step (A) is bonded to a carbon atom forming a triple bond of the monosubstituted alkyne compound (2). And the leaving group X bonded to the carbon atom of the benzene ring of the protected boric acid group-containing benzene compound (3) is removed and the carbon atom to which the hydrogen atom is bonded. And a coupling reaction (A) in which the carbon atoms to which the leaving group X is bonded are bonded (coupled).

  The monosubstituted alkyne compound (2) is a compound having a methoxy group at the 3-position and 5-position of the benzene ring of phenylethyne, as shown in the general formula (2). The monosubstituted alkyne compound (2) can have a methoxy group at the 4-position. The monosubstituted alkyne compound (2) is, for example, anhydrous methylene chloride in which carbon tetrabromide and triphenylphosphine are dissolved in a benzaldehyde compound such as 3,4,5-trimethoxybenzaldehyde or 3,5-dimethoxybenzaldehyde. In the reaction at 0 ° C. for 10 minutes, and 1,1-dibromo-2- (3 ′, 4 ′, 5′-trimethoxyphenyl) ethylene or 1,1-dibromo-2- (3 ′, 5 ′) After obtaining -dimethoxyphenyl) ethylene, it is obtained by further reacting with butyllithium in an anhydrous tetrahydrofuran solvent at -78 ° C.

  The protected boric acid group-containing benzene compound (3) is a compound having a protected boric acid group at either the 3-position or 4-position of the benzene compound as shown in the general formula (3). Further, the protected boric acid group-containing benzene compound (3) can have a methoxy group at the 3-position or the 4-position. In the general formula (3), the leaving group for X is not particularly limited as long as it is a leaving group generally used in organic synthesis reactions, but a halogen atom such as a chlorine atom, a bromine atom, or an iodine atom. A sulfonic acid ester group such as a tosyl group and a triflate group is preferable because it is easily eliminated and a coupling reaction easily occurs. The method for producing the protected boric acid group-containing benzene compound (3) is not particularly limited, and for example, reacting phenylboric acid having a leaving group with pinacol, more specifically, for example, 4-iodo The protected boric acid group-containing benzene compound (3) can be produced by dissolving phenylboric acid and pinacol in methylene chloride, further adding anhydrous sodium sulfate, and reacting for about 12 hours.

  In the present invention, the protected boric acid group is a functional group that is converted into a boric acid group by performing the deprotection step, and the boric acid group has a protective group bonded thereto. Refers to the functional group. The protective boric acid group is not particularly limited. For example, a pinacolate borate group in which a boric acid group is protected using pinacol as a protective group, and a boric acid group is protected using catechol as a protective group. Catecholate borate group, pinandiolate borate group in which boric acid group is protected using pinanediol, 2,3-butanediolate borate group in which boric acid group is protected using 2,3-butanediol, 2,4-pentanediolate borate group protected with 2,4-pentanediol, borate group protected with 1,3-diphenyl-1,3-propanediol Examples include 1,3-diphenyl-1,3-propanediolate borate group.

  In the coupling reaction (A) according to the coupling step (A), a reaction agent is used to advance the reaction of the monosubstituted alkyne compound (2) and the protected boric acid group-containing benzene compound (3). Can do. The reactant is not particularly limited, and examples thereof include triethylamine, tributylamine, triisopropylamine, pyridine, piperazine, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like. Moreover, a catalyst can be used as needed. Although it does not restrict | limit especially as this catalyst, For example, tetrakis-triphenylphosphine palladium and monovalent copper (I) iodide cocatalyst; dibenzylideneacetone palladium, dichlorobistriphenylphosphine palladium, palladium acetate, palladium chloride, etc. Palladium catalysts; phosphine ligands such as triphenylphosphine, tricyclohexylphosphine, tributylphosphine, bisdiphenylphosphinomethane, bisdiphenylphosphinoethane, bisdiphenylphosphinopropane, bisdiphenylphosphinobutane, bisdiphenylphosphinoferrocene, and Examples thereof include a monovalent copper (I) iodide cocatalyst.

  The solvent in the coupling reaction (A) is appropriately selected depending on the reactant and the catalyst. Moreover, as this solvent, although a commercially available thing can be used, the thing from which the water | moisture content was removed by drying operation or dehydration operation is preferable at the point from which reaction efficiency increases.

  The reaction temperature of the coupling reaction (A) is usually 20 to 70 ° C. When the reaction temperature is less than 20 ° C., the reaction does not proceed easily, and when it exceeds 70 ° C., side reactions tend to occur. The reaction time for the coupling reaction (A) is usually 1 to 24 hours. In addition, it is preferable to perform the coupling reaction (A) in an atmosphere of an inert gas such as argon gas using a reaction apparatus that has been dried in advance from the viewpoint of increasing the reaction efficiency.

  Further, the protected boric acid group-containing disubstituted alkyne compound (10) obtained by the coupling reaction (A) can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  An example of the coupling reaction (A) will be described. The monosubstituted alkyne compound (2) and the protected boric acid group-containing benzene compound (3) are dissolved in an acetonitrile solvent, and further, copper (I) iodide and Add tetrakis-triphenylphosphine palladium and stir. Next, a reaction is performed by adding triethylamine to these mixtures. And after predetermined time progress, ammonium chloride aqueous solution is added, reaction is complete | finished, The produced | generated said substituted boric acid group containing disubstituted alkyne compound (10) is extracted using a methylene chloride, An anhydrous magnesium sulfate is extracted. After dehydrating with filtration and the like and filtering, the solvent of the filtrate is removed to obtain the protected boric acid group-containing disubstituted alkyne compound (10).

  The hydrogenation in the hydrogenation step is a stereoselective hydrogenation that converts a triple bond of the protected boric acid group-containing disubstituted alkyne compound (10) into a cis isomer double bond.

  The hydrogenation is not particularly limited as long as the triple bond is stereoselectively converted into a cis isomer, and a method using a hydrogenating agent capable of performing stereoselective hydrogenation, stereoselective hydrogen For example, a method using a hydrogenation catalyst that can perform the conversion is included.

  The solvent for the hydrogenation is appropriately selected depending on the hydrogenating agent or the hydrogenation catalyst. Moreover, as this solvent, although a commercially available thing can be used, the thing from which the water | moisture content was removed by drying operation or dehydration operation is preferable at the point from which reaction efficiency increases.

  The hydrogenation temperature at which the hydrogenation is performed is not particularly limited, but is preferably −10 to 25 ° C., particularly preferably −5 to 5 ° C. When the hydrogenation temperature is less than −10 ° C., hydrogenation hardly occurs, and when it exceeds 25 ° C., stereoselectivity is deteriorated. The hydrogenation time for performing the hydrogenation is not particularly limited, but is generally 1 to 5 hours. If the hydrogenation time is less than 1 hour, the hydrogenation does not occur sufficiently, and if it exceeds 5 hours, the hydrogenation does not proceed any further, which is inefficient. In addition, it is preferable to perform the reaction in an atmosphere of an inert gas such as argon gas using a reaction apparatus that has been dried in advance, from the viewpoint of increasing the reaction efficiency.

  Further, the protected boric acid group-containing cis-stilbene compound (11) obtained by the hydrogenation can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  An example of the hydrogenation method will be described. First, a dicyclohexylborane complex is prepared by reacting a borane-dimethylsulfide complex and cyclohexene in a tetrahydrofuran solvent.

  Next, to the tetrahydrofuran solution of the dicyclohexylborane complex, the protected boric acid group-containing disubstituted alkyne compound (10) is added and reacted at −5 to 5 ° C. for 2 to 4 hours. The reaction is carried out for 0.4 to 0.6 hours. Next, the reaction system is set to -5 to 5 ° C, acetic acid is added, and the reaction is further performed for 1.5 to 3 hours.

  Thereafter, an aqueous sodium hydrogen carbonate solution was added to terminate the hydrogenation, and the resulting protected boric acid group-containing cis-stilbene compound (11) was extracted using methylene chloride, and the extract was dehydrated with anhydrous magnesium sulfate or the like. Then, after filtration, the solvent of the filtrate is removed to obtain the protected boric acid group-containing cis-stilbene compound (11).

  The protected boric acid group-containing cis-stilbene compound (11) obtained by the hydrogenation step has a trans isomer content ratio of 20% or less, preferably 15% or less, particularly preferably 5% or less.

  The deprotection step is a so-called deprotection step in which the protective group of the protected boric acid group-containing cis-stilbene compound (11) is removed and converted to a boric acid group.

  The method for performing the deprotection is not particularly limited, and may be any method generally used for removing the protecting group of the boric acid group. For example, a method using diethanolamine and hydrochloric acid, potassium fluoride and Examples include a method using hydrochloric acid.

  Further, the boric acid group-containing cis-stilbene compound (1) obtained by the deprotection step can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  The boric acid group-containing cis-stilbene compound (1) thus obtained has a trans isomer content ratio of 0.1 to 5% and a purity of 95 to 99.9%. Therefore, the method for producing a boric acid group-containing cis-stilbene compound of the present invention is suitably used for the production of the boric acid group-containing cis-stilbene compound (1) of the present invention.

  Next, a method for producing the boric acid group-containing trans-stilbene compound (A) of the present invention and the boric acid group-containing trans-stilbene compound (B) of the present invention will be described with reference to the following reaction formula (14).

  The method for producing the boric acid group-containing trans-stilbene compound (A) of the present invention includes a toluene derivative represented by the general formula (6) (hereinafter also referred to as a toluene derivative (6)) and the general formula (7). The benzaldehyde derivative shown (hereinafter also referred to as benzaldehyde derivative (7)) is reacted to obtain a leaving group-containing trans-stilbene compound (13), which is obtained in the coupling step (B). A boric acid group introducing step of obtaining a boric acid group-containing trans-stilbene compound (A) by replacing the leaving group of the leaving group-containing trans-stilbene compound (13) with a boric acid group.

  Moreover, the manufacturing method of the boric acid group containing trans-stilbene compound (B) of this invention makes this toluene derivative (6) and this benzaldehyde derivative (7) react, and a leaving group containing trans-stilbene compound (13) is made. The leaving group of the leaving group-containing trans-stilbene compound (13) obtained in the coupling step (B) and the coupling step (B) is substituted with a boric acid group, and the boric acid group-containing trans-stilbene is obtained. A boric acid group introduction step for obtaining a compound (A), a methoxy group of the boric acid group-containing trans-stilbene compound (A) obtained in the boric acid group introduction step is converted into a hydroxyl group, and a boric acid group-containing trans A hydroxylation step for obtaining a stilbene compound (B);

  The reaction of the toluene derivative (6) and the benzaldehyde derivative (7) in the coupling step (B) involves the reaction of the carbon atom to which the phosphonium group or phosphate group of the toluene derivative (6) is bonded with the benzaldehyde. Coupling reaction (B) in which the carbon atom of the carbonyl group of the derivative (7) is bonded, and the phosphonium group or phosphate group and the oxygen atom of the carbonyl group are eliminated to form a double bond. And a reaction for stereoselectively forming a trans stilbene skeleton.

As shown in the general formula (6), the toluene derivative (6) is 2 or 3 of 2-position, 3-position, 4-position, 5-position and 6-position of the benzene ring of the toluene derivative. It has a methoxy group at the site. In the general formula (6), Y represents a phosphonium halide group (—P ⁺ R ₃ X ⁻ ), a phosphate group (—PO (OH) ₂ ), or a phosphate ester group (—PO (OR) OH). Or -PO (OR) ₂ ). Examples of the phosphonium halide group include a triphenylphosphonium bromide group. The toluene derivative (6) is produced, for example, by the method described in Non-Patent Document, J. Org. Chem. 62, 4821-4826 (1997) (Meier, H .; Dullweber, U). . Specifically, for example, it is obtained by reacting 3,5-dimethoxybenzyl bromide and triethyl phosphite (P (OEt) ₃ ) at 160 ° C. for 12 hours without solvent.

  The benzaldehyde derivative (7) is a compound having a methoxy group at the 4-position of benzaldehyde as shown in the general formula (7). In the general formula (7), the leaving group for Z is not particularly limited as long as it is a leaving group generally used in organic synthesis reaction, but halogen atoms such as chlorine atom, bromine atom, iodine atom, etc. However, since it is easy to lithiate, it is preferable at the point which is easy to substitute with a boric acid group. The method for producing the benzaldehyde derivative (7) is not particularly limited. For example, the benzaldehyde derivative (7) can be produced by reacting 4-anisaldehyde with a halogen molecule such as bromine.

  In the coupling reaction (B) according to the coupling step (B), a reactant can be used in order to advance the reaction of the toluene derivative (6) and the benzaldehyde derivative (7). The reactant is not particularly limited, and examples thereof include tert-butoxy potassium, sodium hydride, potassium hydride, lithium hydride, n-butyl lithium, lithium diisopropylamide, sodium methoxide and the like.

  The solvent in the coupling reaction (B) is appropriately selected depending on the reactant. Moreover, as this solvent, although a commercially available thing can be used, the thing from which the water | moisture content was removed by drying operation or dehydration operation is preferable at the point from which reaction efficiency increases.

  The reaction temperature of the coupling reaction (B) is usually 0 to 25 ° C. When the reaction temperature is less than 0 ° C., the reaction hardly proceeds, and when the reaction temperature exceeds 25 ° C., side reactions tend to occur. The reaction time for the coupling reaction (B) is usually 2 to 12 hours. In addition, it is preferable to perform the coupling reaction (B) in an atmosphere of an inert gas such as argon gas using a reaction apparatus that has been dried in advance from the viewpoint of increasing the reaction efficiency.

  Further, the leaving group-containing trans-stilbene compound (13) obtained by the coupling reaction (B) can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  To explain an example of the coupling reaction (B), the toluene derivative (6) and the benzaldehyde derivative (7) are dissolved in a dimethylformamide solvent, and tert-butoxypotassium dissolved in dimethylformamide is further added and stirred. , React. Then, after a predetermined time has elapsed, the resulting leaving group-containing trans-stilbene compound (13) is extracted using diethyl ether, the extract is dehydrated with anhydrous sodium sulfate and the like, filtered, and then the solvent of the filtrate. Is removed, the leaving group-containing trans-stilbene compound (13) can be obtained.

  In the leaving group-containing trans-stilbene compound (13) obtained by the coupling step (B), the content ratio of the cis isomer to the trans isomer is 10% or less, preferably 1% or less, particularly preferably 0.5%. It is as follows.

  The boric acid group introduction step is a step of performing a substitution reaction in which the leaving group Z of the leaving group-containing trans-stilbene compound (13) is substituted with a boric acid group.

In the substitution reaction, the boric acid group introducing agent for substituting the leaving group Z with a boric acid group is not particularly limited as long as it is usually used to substitute the leaving group with a boric acid group. Examples thereof include alkyl borate (B (OR) ₃ ) such as triethyl borate and triisopropyl borate; borane halide (BH ₂ X) and the like.

  In the substitution reaction, a reactive agent can be used to advance the reaction. Although it does not restrict | limit especially as this reactant, For example, n-bryl lithium, sec-butyl lithium, t-butyl lithium etc. are mentioned. Moreover, a catalyst can be used as needed. The catalyst is not particularly limited, and examples thereof include tetramethylethylenediamine (TMEDA), dimethylformamide (DMF), hexamethylphosphorylamide (HMPA), and hexamethylphosphoryltriamide (HMPT).

  The solvent in the substitution reaction is appropriately selected depending on the boric acid group introducing agent, the reactant, or the catalyst. Moreover, as this solvent, although a commercially available thing can be used, the thing from which the water | moisture content was removed by drying operation or dehydration operation is preferable at the point from which reaction efficiency increases.

  The reaction temperature for carrying out the substitution reaction is not particularly limited, but is preferably -100 to -20 ° C, particularly preferably -80 to -60 ° C. When the reaction temperature is less than −100 ° C., the substitution reaction hardly occurs, and when it exceeds −20 ° C., the production of a reduced form (a compound in which the leaving group Z is substituted with a hydrogen atom) increases. The reaction time for carrying out the substitution reaction is not particularly limited, but is generally 0.5 to 1 hour. If the reaction time is less than 0.5 hours, the substitution reaction does not occur sufficiently, and if the reaction time exceeds 1 hour, the substitution reaction does not proceed any further, which is inefficient. In addition, it is preferable to perform the reaction in an atmosphere of an inert gas such as argon gas using a reaction apparatus that has been dried in advance, from the viewpoint of increasing the reaction efficiency.

  Further, the boric acid group-containing trans-stilbene compound (A) obtained by the substitution reaction can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  To explain an example of the substitution reaction, the leaving group-containing trans-stilbene compound (13) is dissolved in a tetrahydrofuran solvent, and further a toluene solvent is added, followed by cooling to −80 to −60 ° C. Then, isopropyl borate is dropped and stirred for a predetermined time. Next, n-butyllithium is added, and the mixture is stirred at −80 to −60 ° C. for a predetermined time, gradually raised in temperature, and further stirred at 0 to 20 ° C. for a predetermined time.

  Thereafter, an aqueous sulfuric acid solution was added to complete the substitution reaction, and the resulting boric acid group-containing trans-stilbene compound (A) was extracted using methylene chloride, and the extract was dehydrated with anhydrous sodium sulfate and filtered. Then, the boric acid group-containing trans-stilbene compound (A) is obtained by removing the solvent of the filtrate.

  The boric acid group-containing trans-stilbene compound (A) thus obtained has a cis isomer content ratio of 0.1 to 1% and a purity of 99 to 99.9%. Therefore, the method for producing the boric acid group-containing trans-stilbene compound (A) of the present invention is suitably used for the production of the boric acid group-containing trans-stilbene compound (A) of the present invention.

  The hydroxylation step is a step of performing a hydroxylation reaction in which the methoxy group of the boric acid group-containing trans-stilbene compound (A) is converted to a hydroxyl group.

  The method for performing the hydroxylation reaction is not particularly limited, and may be any method generally used for converting a methoxy group into a hydroxyl group. For example, a method using boron tribromide, boron trichloride , A method using trimethylsilane iodide, a method using sodium ethanethiolate, and the like.

  The solvent in the hydroxylation reaction is appropriately selected. Moreover, as this solvent, although a commercially available thing can be used, the thing from which the water | moisture content was removed by drying operation or dehydration operation is preferable at the point from which reaction efficiency increases.

  The reaction temperature of the hydroxylation reaction is usually −80 to −20 ° C. When the reaction temperature is less than −80 ° C., the reaction hardly proceeds, and when it exceeds −20 ° C., side reactions are liable to occur. The reaction time of the hydroxylation reaction is usually 6 to 12 hours. In addition, it is preferable to perform the hydroxylation reaction in an atmosphere of an inert gas such as argon gas using a reaction apparatus that has been dried in advance from the viewpoint of increasing the reaction efficiency.

  Further, the boric acid group-containing trans-stilbene compound (B) obtained by the hydroxylation step can be purified by silica gel chromatography, recrystallization or the like, if necessary.

  The boric acid group-containing trans-stilbene compound (B) thus obtained has a cis isomer content ratio of 0.1 to 1% and a purity of 99 to 99.9%. Therefore, the method for producing the boric acid group-containing trans-stilbene compound (B) of the present invention is suitably used for the production of the boric acid group-containing trans-stilbene compound (B) of the present invention.

  EXAMPLES Next, although an Example is given and this invention is demonstrated more concretely, this is only an illustration and does not restrict | limit this invention.

(Coupling step (A))
2.9 g (15 mmol) of 3,4,5-trimethoxyphenylethyne (a compound in which R ¹ is a methoxy group in the general formula (2)) and 4-methoxy-3- (pinaco) at room temperature under an argon atmosphere (Rate borate) Bromobenzene (in the general formula (3), R ⁵ is a pinacolate borate group, R ⁶ is a methoxy group, and X is a bromine atom) 4.7 g (15 mmol) was dissolved in dehydrated acetonitrile solvent. . Next, 0.22 g (1.2 mmol) of copper chloride (I) (CuI) and 0.87 g (0.75 mmol) of tetrakis-triphenylphosphine palladium (Pd (PPh ₃ ) ₄ ) were added to the acetonitrile solution. . Next, 6.3 ml (45 mmol) of triethylamine was added with stirring, and the mixture was stirred at 70 ° C. for 20 hours to carry out a coupling reaction.

  After stirring for a predetermined time, an aqueous ammonium chloride solution was added to terminate the coupling reaction, and extraction was performed using methylene chloride. The organic layer was dehydrated with a saturated aqueous sodium chloride solution, further dried over anhydrous magnesium sulfate, and then filtered. The solvent in the obtained filtrate was removed by an evaporator to obtain a yellowish white solid. The yellowish white solid was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 4/1) to give a pale yellow solid of 3,4,5-trimethoxyphenyl-4′-methoxy-3 ′-( 1.2 g of pinacolate borate) phenylethyne (hereinafter referred to as protected boric acid group-containing disubstituted alkyne compound (D1)) was obtained. At this time, the yield was 20%. Table 1 shows the physical properties of the protected boric acid group-containing disubstituted alkyne compound (D1).

(Hydrogenation process)
0.61 ml (6.0 mmol) of cyclohexene was dissolved in 8.0 ml of tetrahydrofuran, and the tetrahydrofuran solution was brought to 0 ° C. Next, 3.0 ml (3.0 mmol) of a 1M solution of borane-dimethylsulfide complex was added dropwise to the tetrahydrofuran solution over 1 minute, followed by stirring for 1 hour, followed by the protection boron dissolved in 12 ml of tetrahydrofuran. 1.1 g (2.5 mmol) of the acid group-containing disubstituted alkyne compound (D1) was added, stirred at 0 ° C. for 2.5 hours, and further stirred at room temperature for 30 minutes. Thereafter, the tetrahydrofuran solution was again brought to 0 ° C., 1.0 ml (17 mmol) of acetic acid was added and the mixture was stirred for 2 hours. Next, a saturated aqueous sodium hydrogen carbonate solution was added, followed by extraction with methylene chloride. The organic layer was dehydrated with an aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and then filtered. The solvent in the obtained filtrate was removed, and the residue was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 4: 1) to obtain 3,4,4 ′, 5-tetramethoxy-3 ′-( 0.44 g of pinacolate borate) cis-stilbene (hereinafter referred to as protected boric acid group-containing cis-stilbene compound (E1)) was obtained. At this time, the yield was 40%. Table 2 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (E1).

(Deprotection process)
Diethanolamine (0.12 g, 1.1 mmol) dissolved in 2-propanol (0.3 ml) and the protected boric acid group-containing cis-stilbene compound (E1) (0.44 g, 1.0 mmol) were mixed, followed by diethyl ether 3 0.0 ml was added and stirred at room temperature for 48 hours. After stirring for a predetermined time, filtration was performed, and the solid separated by filtration was washed with diethyl ether to obtain a white solid. The white solid was dissolved in 3.0 ml of tetrahydrofuran, 3.0 ml of 1 mol / L hydrochloric acid aqueous solution was added, and the mixture was stirred at room temperature for 1 hour. After stirring, distilled water and ethyl acetate were added for extraction. The organic layer was brought into contact with a saturated aqueous sodium hydrogen carbonate solution to make it weakly basic, then dehydrated with a saturated aqueous sodium chloride solution, and further dried over anhydrous magnesium sulfate. After filtration, the solvent in the filtrate was removed and the residue was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 3/1) to obtain 3,4,4 ′, 5-tetramethoxy-3 as a white solid. '-(Dihydroxylboryl) cis-stilbene (compound in which R ¹ is a methoxy group, R ² is a boric acid group, and R ³ is a methoxy group in the general formula (1), hereinafter referred to as a boric acid group-containing cis-stilbene compound) (Described as (C1).) 0.35 g was obtained. At this time, the yield was 10%. Table 3 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (C1).

(Water solubility of boric acid group-containing cis-stilbene compound)
The solubility of the boric acid group-containing cis-stilbene compound (C1) in water was 2.36 × 10 ⁻⁴ mol / L at 23 ° C. For comparison, the solubility of combretastatin A-4 was determined and found to be 3.86 × 10 ⁻⁵ mol / L at 23 ° C.

(Evaluation of physiological activity of boric acid group-containing cis-stilbene compound)
(I) Evaluation of cell growth inhibitory activity against B16 cells Boric acid group-containing cis-stilbene compound (C1) was dissolved in dimethyl sulfoxide. Next, to a 96-well plate “3072” (manufactured by Falcon), 0.5 × 10 ⁴ B16 cells, 200 μL of medium and the dimethyl sulfoxide solution are added per well, and the content of dimethyl sulfoxide is finally 0. The boric acid group-containing cis-stilbene compound (C1) was adjusted to have a concentration of 0.01 to 100 μg / mL. This is cultured at 37 ° C. in a 5% carbon dioxide atmosphere for 3 days, and then the medium is removed and the plate is washed 3 times with 50 μL of a phosphate buffer solution PBS (−). Cells were stained with 20 μL of 0.4% methanol solution of crystal violet, and absorbance was measured with a microplate reader (wavelength 490 nm) to determine 50% cell growth inhibitory concentration (IC ₅₀ ). As a result, the cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C1) with respect to B16 cells was 6.3 × 10 ⁻⁹ mol / L.
(Ii) Evaluation of cell growth inhibitory activity against 1-87 cells The above (i) cells against B16 cells except that 0.5 × 10 ⁴ B16 cells are used instead of 0.5 × 10 ⁴ cells As a result of performing the same method as the evaluation of the growth inhibitory activity, the cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C1) with respect to 1-87 cells was 5.1 × 10 ⁻⁹ mol / L. Met.

(Evaluation of physiological activity of combretastatin A-4)
For comparison, instead of the boric acid group-containing cis-stilbene compound (C1), the compound represented by the above formula (16) in which the boric acid group of the boric acid group-containing cis-stilbene compound (C1) is a hydroxyl group is used. The physiological activity was evaluated in the same manner as described above except that lettastatin A-4 was used. The cell growth inhibitory concentration (IC50) of combretastatin A-4 against B16 cells was 4.6 × 10 ^{−. 9} is a mol / L, cell growth inhibitory concentration of combretastatin a-4 for 1-87 cells (IC50) was 8.5 × ¹⁰ -9 mol / L.

  Thus, it was found that the boric acid group-containing cis-stilbene compound (C1) is about 10 times more soluble in water than combretastatin A-4 and has tubulin polymerization inhibitory activity.

(Coupling step (A))
Under an argon atmosphere at room temperature, 0.74 g (3.8 mmol) of 3,4,5-trimethoxyphenylethyne and 4- (pinacolate borate) iodobenzene (in the general formula (3), R ⁵ represents a hydrogen atom. , R ⁶ is a pinacolate borate group, X is an iodine atom compound) 1.2 g (3.8 mmol) was dissolved in dehydrated acetonitrile solvent. Next, 0.072 g (0.038 mmol) of copper chloride (I) (CuI) and 0.22 g (0.019 mmol) of tetrakis-triphenylphosphine palladium (Pd (PPh ₃ ) ₄ ) were added to the acetonitrile solution. . Next, 0.93 ml (6.7 mmol) of triethylamine was added with stirring, and the mixture was stirred at 70 ° C. for 1.5 hours to carry out a coupling reaction.

  After stirring for a predetermined time, a saturated aqueous ammonium chloride solution was added to terminate the coupling reaction, and extraction was performed using methylene chloride. The organic layer was dehydrated with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and then filtered. The solvent in the obtained filtrate was removed by an evaporator to obtain a yellowish white solid. The yellowish white solid was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 10/1) to give 3,4,5-trimethoxyphenyl-4 ′-(pinacolate borate) phenyl as a pale yellow solid. 1.1 g of ethyne (hereinafter referred to as a protected boric acid group-containing disubstituted alkyne compound (D2)) was obtained. At this time, the yield was 71%. Table 4 shows the physical properties of the protected boric acid group-containing disubstituted alkyne compound (D2).

(Hydrogenation process)
Example except that 1.1 g (2.5 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D1) was replaced with 0.59 g (1.5 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D2). 1 was carried out in the same manner as in No. 1, but the oily liquid 3,4,5-trimethoxy-4 ′-(pinacolatoborate) cis-stilbene (hereinafter referred to as protected boric acid group-containing cis-stilbene compound (E2) is also described. .) 0.40 g was obtained. At this time, the yield was 67%. Table 5 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (E2).

(Deprotection process)
The protected boric acid group-containing cis-stilbene compound (E2) is dissolved in 3.0 ml of methanol, and 0.5 ml (2.0 mmol) of a 4 mol / L potassium hydrogen fluoride aqueous solution is added to the obtained methanol solution, and then at room temperature for 30 minutes. Stir. After stirring, filtration was performed, and the filtered solid was washed with diethyl ether to obtain a white solid. The white solid was dissolved in 3.0 ml of ethyl acetate, 3.0 ml of 1 mol / L hydrochloric acid aqueous solution was added, and the mixture was stirred at room temperature for 1 hour. The organic layer obtained by removing the aqueous layer was brought into contact with a saturated aqueous sodium hydrogen carbonate solution to make it weakly basic, then dehydrated with a saturated aqueous sodium chloride solution, and further dried over anhydrous magnesium sulfate. After filtration, the solvent in the obtained filtrate was removed, recrystallized using hexane-methylene chloride, and 3,4,5-trimethoxy-4 ′-(dihydroxylboryl) cis-stilbene (the above general formula) In (1), R ¹ is a methoxy group, R ² is a hydrogen atom, and R ³ is a boric acid group, hereinafter referred to as a boric acid group-containing cis-stilbene compound (C2).) 0.054 g was obtained. . At this time, the yield was 46%. Table 6 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (C2).

(Evaluation of physiological activity of boric acid group-containing cis-stilbene compound)
(I) Evaluation of cell growth inhibitory activity against B16 cells The same method as in Example 1, except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C2) with respect to B16 cells was 2.6 × 10 ⁻⁶ mol / L.
(Ii) Evaluation of cell growth inhibitory activity against 1-87 cells As in Example 1, except that the boric acid group-containing cis-stilbene compound (C1) was used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C2) with respect to 1-87 cells was 3.2 × 10 ⁻⁶ mol / L.

(Coupling step (A))
Instead of 1.2 g (3.8 mmol) of 4- (pinacolate borate) iodobenzene, 3- (pinacolate borate) iodobenzene (in the general formula (3), R ⁵ is a pinacolate borate group, and R ⁶ is hydrogen. Except for 2.0 g (6.0 mmol) of a compound having an atom and X being an iodine atom), a yellowish white solid of 3,4,5-trimethoxyphenyl-3 was obtained in the same manner as in Example 2. 1.7 g of '-(pinacolate borate) phenylethyne (hereinafter referred to as a protected boric acid group-containing disubstituted alkyne compound (D3)) was obtained. At this time, the yield was 72%. Table 7 shows the physical properties of the protected boric acid group-containing disubstituted alkyne compound (D3).

(Hydrogenation process)
Example except that 1.1 g (2.5 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D1) was replaced with 1.2 g (3.1 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D3). 1 was carried out in the same manner as in No. 1, and was described as an oily liquid 3,4,5-trimethoxy-3 ′-(pinacolate borate) cis-stilbene (hereinafter referred to as a protected boric acid group-containing cis-stilbene compound (E3)). .) 0.6 g was obtained. At this time, the yield was 53%. Table 8 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (E3).

(Deprotection process)
Except for replacing the protected boric acid group-containing cis-stilbene compound (E1) 0.44 g (1.0 mmol) with the protected boric acid group-containing cis-stilbene compound (E3) 0.82 g (2.1 mmol), When carried out in the same manner as in Example 1, 3,4,5-trimethoxy-3 ′-(dihydroxylboryl) cis-stilbene (in the general formula (1), R ¹ is a methoxy group, R A compound in which ² is a boric acid group and R ³ is a hydrogen atom, hereinafter referred to as a boric acid group-containing cis-stilbene compound (C3). At this time, the yield was 29%. Table 9 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (C3).

(Evaluation of physiological activity of boric acid group-containing cis-stilbene compound)
(I) Evaluation of cell growth inhibitory activity against B16 cells The same method as in Example 1, except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C3) with respect to B16 cells was 4.9 × 10 ⁻⁷ mol / L.
(Ii) Evaluation of cell growth inhibitory activity against 1-87 cells As in Example 1, except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C3) with respect to 1-87 cells was 2.1 × 10 ⁻⁶ mol / L.

(Coupling step (A))
0.88 g (5.4 mmol) of 3,5-dimethoxyphenylethyne (a compound in which R ¹ is a hydrogen atom in the general formula (2)) and 4- (pinacolate borate) iodobenzene at room temperature under an argon atmosphere 1.8 g (5.4 mmol) was dissolved in dehydrated acetonitrile solvent. Next, 0.10 g (0.54 mmol) of copper (I) chloride (CuI) and 0.31 g (0.27 mmol) of tetrakis-triphenylphosphine palladium (Pd (PPh ₃ ) ₄ ) were added to the acetonitrile solution. . Next, 2.2 ml (16 mmol) of triethylamine was added with stirring, and the mixture was stirred at 50 ° C. for 2 hours to carry out a coupling reaction.

  After stirring for a predetermined time, an aqueous ammonium chloride solution was added to terminate the coupling reaction, and extraction was performed using methylene chloride. The organic layer was dehydrated with a saturated aqueous sodium chloride solution, further dried over anhydrous magnesium sulfate, and then filtered. The solvent in the obtained filtrate was removed by an evaporator to obtain a yellowish white solid. The yellowish white solid was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 30/1) to give 3,5-dimethoxyphenyl-4 ′-(pinacolatoborate) phenylethyne (hereinafter referred to as “yellowish white solid”). , Described as a protected boric acid group-containing disubstituted alkyne compound (D4).) 1.4 g was obtained. At this time, the yield was 73%. Table 10 shows the physical properties of the protected boric acid group-containing disubstituted alkyne compound (D4).

(Hydrogenation process)
Example 1 except that 1.1 g (2.5 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D1) is replaced with 0.80 g (2.2 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D4). When conducted in the same manner as in No. 1, 3,5-dimethoxy-4 ′-(pinacolatoborate) cis-stilbene (hereinafter referred to as a protected boric acid group-containing cis-stilbene compound (E4)). 0.55 g was obtained. At this time, the yield was 68%. Table 11 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (E4).

(Deprotection process)
Example except that 0.4 g (1.0 mmol) of the protected boric acid group-containing cis-stilbene compound (E1) was used instead of 0.5 g (1.4 mmol) of the protected boric acid group-containing cis-stilbene compound (E4) 1 was carried out in the same manner as in Example 1. As a result, white solid 3,5-dimethoxy-4 ′-(dihydroxylboryl) cis-stilbene (in the general formula (1), R ¹ was a hydrogen atom and R ² was a hydrogen atom). , R ³ is a compound having a boric acid group, hereinafter referred to as a boric acid group-containing cis-stilbene compound (C4).) 0.1 g was obtained. At this time, the yield was 25%. Table 12 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (C4).

(Evaluation of physiological activity of boric acid group-containing cis-stilbene compound)
(I) Evaluation of cell growth inhibitory activity against B16 cells The same method as in Example 1 except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C4) for B16 cells was 1.2 × 10 ⁻⁵ mol / L.
(Ii) Evaluation of cell growth inhibitory activity against 1-87 cells As in Example 1, except that the boric acid group-containing cis-stilbene compound (C1) was used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C4) for 1-87 cells was 9.1 × 10 ⁻⁶ mol / L.

(Coupling step (A))
In the same manner as in Example 4, except that 1.8 g (5.4 mmol) of 4- (pinacolate borate) iodobenzene was used and 1.3 g (4.0 mmol) of 3- (pinacolate borate) iodobenzene was used. As a result, 1.0 g of 3,5-dimethoxyphenyl-3 ′-(pinacolate borate) phenylethyne (hereinafter referred to as a protected boric acid group-containing disubstituted alkyne compound (D5)) was obtained. It was. At this time, the yield was 70%. Table 13 shows the physical properties of the protected boric acid group-containing disubstituted alkyne compound (D5).

(Hydrogenation process)
Example 1 except that 1.1 g (2.5 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D1) is replaced with 0.88 g (2.4 mmol) of the protected boric acid group-containing disubstituted alkyne compound (D5). When conducted in the same manner as in No. 1, 3,5-dimethoxy-3 ′-(pinacolate borate) cis-stilbene (hereinafter referred to as a protected boric acid group-containing cis-stilbene compound (E5)). 0.41 g was obtained. At this time, the yield was 47%. Table 14 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (E5).

(Deprotection process)
Example except that 0.44 g (1.0 mmol) of the protected boric acid group-containing cis-stilbene compound (E1) was used instead of 0.33 g (0.89 mmol) of the protected boric acid group-containing cis-stilbene compound (E5) 1 was carried out in the same manner as in Example 1. As a result, white solid 3,5-dimethoxy-3 ′-(dihydroxylboryl) cis-stilbene (in the general formula (1), R ¹ was a hydrogen atom and R ² was boric acid). 0.19 g of a group, a compound in which R ³ is a hydrogen atom, hereinafter referred to as a boric acid group-containing cis-stilbene compound (C5). At this time, the yield was 68%. Table 15 shows the physical properties of the protected boric acid group-containing cis-stilbene compound (C5).

(Evaluation of physiological activity of boric acid group-containing cis-stilbene compound)
(I) Evaluation of cell growth inhibitory activity against B16 cells The same method as in Example 1 except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the boric acid group-containing cis-stilbene compound (C5) with respect to B16 cells was 1.8 × 10 ⁻⁶ mol / L.
(Ii) Evaluation of cell growth inhibitory activity against 1-87 cells The same as Example 1 except that the boric acid group-containing cis-stilbene compound (C1) is used instead of the boric acid group-containing cis-stilbene compound (C1). The cell growth inhibitory concentration (IC50) of the borate group-containing cis-stilbene compound (C5) with respect to 1-87 cells was 2.0 × 10 ⁻⁶ mol / L.

(Coupling step (B))
3,5-dimethoxybenzyl phosphoric acid diethyl ester (in general formula (6), n is 2, Y is phosphoric acid diethyl ester group (—PO (OEt) ₂ )) And a compound having a methoxy group at the 5-position) 1.44 g (5 mmol), and 3-bromo-4-methoxybenzaldehyde (a compound in which Z is a bromine atom in the general formula (7)) 1.08 (5 mmol) Was dissolved in 20 ml of dehydrated dimethylformamide solvent. Next, 2.00 g (15 mmol) of tert-butoxy potassium dissolved in 10 ml of dehydrated dimethylformamide solvent was added dropwise at 0 ° C. and stirred at room temperature for 12 hours to carry out a coupling reaction. And after stirring for 12 hours, it extracted using diethyl ether.

  Water was added to the diethyl ether layer to remove dimethylformamide, washed with a 5% sodium hydrogen carbonate solution, dehydrated with a saturated aqueous sodium chloride solution, further dried over sodium sulfate, and filtered. The solvent in the obtained filtrate was removed with an evaporator to obtain a yellow solid.

  The yellow solid was purified by silica gel column chromatography (developing solvent: hexane / acetic acid = 10/1) to obtain 3,4 ′, 5-trimethoxy-3′-bromo-trans-stilbene (hereinafter referred to as “dehydrated”) as a yellowish white solid. It is described as a leaving group-containing trans-stilbene compound (F1).) 0.99 g (13.2 mmol) was obtained. At this time, the yield was 63%. The physical properties of the leaving group-containing trans-stilbene compound (F1) are shown below.

(Boric acid group introduction process)
Under an argon atmosphere, 1.75 g (5.0 mmol) of the leaving group-containing trans-stilbene compound (F1) was dissolved in 5 ml of dehydrated tetrahydrofuran, and further 20 ml of toluene solvent was added to bring the temperature to −78 ° C. Subsequently, 1.74 ml (7.5 mmol) of isopropyl borate (B (iso-C ₃ H ₇ O) ₃ ) was added dropwise and stirred for 30 minutes.

  Subsequently, 4.38 ml (7.0 mmol) of n-butyllithium was added, and the mixture was stirred at -78 ° C for 1 hour, gradually brought to room temperature, and stirred for 1 hour. Then, it was made -20 degreeC, 2 ml of 10% sulfuric acid aqueous solution was added, it was made room temperature, and reaction was terminated.

  Subsequently, extraction was performed using methylene chloride. The organic layer was dehydrated with a saturated aqueous sodium chloride solution, dried using sodium sulfate, and then filtered. The solvent in the obtained filtrate was removed with an evaporator to obtain a yellow solid. The yellow solid was purified by silica gel column chromatography (developing solvent: hexane / ethyl acetate = 4/1), and 3,4 ', 5-trimethoxy-3'-(dihydroxylboryl) trans-stilbene was obtained as a yellowish white solid. (In the formula (4), n is 2, a compound having a methoxy group at the 3-position and the 5-position, hereinafter referred to as a boric acid group-containing trans-stilbene compound (A1)) 0.84 g ( 1.67 mmol) was obtained. At this time, the yield was 53%. The physical properties of the boric acid group-containing trans-stilbene compound (A1) are shown below.

(Hydroxylation step)
Under an argon atmosphere, 0.63 g (2.0 mmol) of the boric acid group-containing trans-stilbene compound (A1) was dissolved in a dehydrated methylene chloride solvent and stirred at -78 ° C. 7.6 ml (6.0 mmol) of a boron bromide solution (BBr ₃ , 0.783 M) was added dropwise thereto and reacted for 1 hour, then the temperature was gradually raised to −40 ° C. and the mixture was further stirred for 8 to 12 hours. The reaction was performed. After the reaction, the solution was dropped into ice water to terminate the reaction, followed by extraction with ethyl acetate. The organic layer was washed with water and dehydrated with a saturated aqueous sodium chloride solution, and then the solvent was removed with an evaporator to obtain a brown solid. The brown solid was purified by silica gel column chromatography (developing solvent: methylene chloride / methanol = 5/1), and the brown solid 3,4 ', 5-trihydroxyl-3'-(dihydroxylboryl) trans-stilbene (In the formula (5), m is 2, a compound having a hydroxyl group at the 3-position and the 5-position, hereinafter referred to as a boric acid group-containing trans-stilbene compound (B1)) 0.41 g ( 1.5 mmol) was obtained. At this time, the yield was 75%. The physical properties of the boric acid group-containing trans-stilbene compound (B1) are shown below. The boric acid group-containing trans-stilbene compound (B1) has a hydroxyl group of a 3′-position dihydroxyboryl group converted to a methoxy group in methanol, which is a solvent for MS spectrum analysis. 4 ', 5-trihydroxyl-3'-(dimethoxyboryl) trans-stilbene was detected.
・ MS (ESI negative) m / z; 299 (C ₁₄ H ₁₀ O ₃ B (OMe) ₂ )

Furthermore, in order to confirm the structure of the compound obtained by hydroxylation, (1S, 2S, 3R, 5S)-(+)-pinanediol was used to convert the compound after hydroxylation and purification into stable boric acid. When the molecular weight of the resulting boric acid ester compound induced by ester was measured by MS (ESI positive), m / z; 407 (MH ⁺ ) was confirmed. From this, it was confirmed that the boric acid ester compound has a structure of the following formula (15) having a boric acid group and all methoxy groups being hydroxylated. Therefore, it was confirmed that the structure of the compound obtained by hydroxylation and purification was 3,4 ', 5-trihydroxyl-3'-(dihydroxylboryl) trans-stilbene.
・ MS (ESI positive) m / z 407 (C ₂₄ H ₂₇ O ₃ BO ₅ )

(Evaluation of physiological activity of boric acid group-containing trans-stilbene compound)
(I) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (A) on B-16 cells Boric acid group-containing trans-stilbene compound (A1) was dissolved in dimethyl sulfoxide. Next, to a 96-well plate “3072” (Falcon), 0.5 × 10 ⁴ B-16 cells per well, 200 μL of medium, and the dimethyl sulfoxide solution were added, and finally the content of dimethyl sulfoxide Of 0.5% or less, and the concentration of the boric acid group-containing trans-stilbene compound (A1) was adjusted to 0.01 to 100 μg / mL. This is cultured at 37 ° C. in a 5% carbon dioxide atmosphere for 3 days, and then the medium is removed and the plate is washed 3 times with 50 μL of a phosphate buffer solution PBS (−). Cells were stained with 20 μL of 0.4% methanol solution of crystal violet, and absorbance was measured with a microplate reader (wavelength 490 nm) to determine 50% cell growth inhibitory concentration (IC ₅₀ ). As a result, the cell growth inhibitory concentration (IC50) of the boric acid group-containing trans-stilbene compound (A1) with respect to B-16 cells was 1.1 × 10 ⁻⁵ mol / L.

(Ii) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (B) against B-16 cells Instead of boric acid group-containing trans-stilbene compound (A1), boric acid group-containing trans-stilbene compound (A1) Except for B1), boric acid for B-16 cells was obtained by the same method as the evaluation of the cell growth inhibitory activity of the boric acid group-containing trans-stilbene compound (A) for B-16 cells. The cell growth inhibitory concentration (IC50) of the group-containing trans-stilbene compound (B1) was 9.0 × 10 ⁻⁶ mol / L.

(Evaluation of physiological activity of resveratrol)
(I) Inhibition of cell growth of boric acid group-containing trans-stilbene compound (A) on B-16 cells except that resveratrol (18) is used instead of boric acid group-containing trans-stilbene compound (A1) When the same method as the evaluation of activity was performed, the cell growth inhibitory concentration (IC50) of resveratrol (18) for B-16 cells was 8.8 × 10 ⁻⁵ mol / L.

(Evaluation of physiological activity of resveratrol derivative)
Instead of the boric acid group-containing trans-stilbene compound (A1), a trimethyl ether form of resveratrol (18) (3,4 ', 5-trimethoxy-trans-stilbene, a compound described in the following formula (19)) and Except that (i) the evaluation of the cell growth inhibitory activity of the boric acid group-containing trans-stilbene compound (A) on B-16 cells was carried out in the same manner as above, and the trimethyl of resveratrol against B-16 cells The cell growth inhibitory concentration (IC50) of the ether derivative (19) was 2.4 × 10 ⁻⁵ mol / L.

(Coupling step (B))
In place of 3,5-dimethoxybenzyl phosphoric acid diethyl ester, 3,4-dimethoxybenzyl phosphoric acid diethyl ester (in the general formula (6), n is 2, Y is phosphoric acid diethyl ester group (-PO ( OEt) ₂ ), which is a compound having a methoxy group at the 3-position and 4-position)) except that 1.44 g (5.0 mmol) is obtained, and is carried out in the same manner as in Example 6, and 3, 4, 4 ′ 0.74 g (2.1 mmol) of -trimethoxy-3′-bromo-trans-stilbene (hereinafter referred to as a leaving group-containing trans-stilbene compound (F2)) was obtained in a yield of 42%. The physical properties of the leaving group-containing trans-stilbene compound (F2) are shown below.

(Boric acid group introduction process)
The same procedure as in Example 6 was performed except that 1.05 g (3.0 mmol) of the leaving group-containing trans-stilbene compound (F2) was used instead of the leaving group-containing trans-stilbene compound (F1). As a result, 3,4,4′-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene (in the above formula (4), n is 2 and methoxy at the 3-position and 4-position) A compound having a group, hereinafter referred to as a boric acid group-containing trans-stilbene compound (A2).) 0.74 g (2.3 mmol) was obtained. At this time, the yield was 78%. The physical properties of the boric acid group-containing trans-stilbene compound (A2) are shown below.

(Hydroxylation step)
The same procedure as in Example 6 was performed except that the boric acid group-containing trans-stilbene compound (A1) was replaced with 0.27 g (0.86 mmol) of the boric acid group-containing trans-stilbene compound (A2). As a result, 3,4,4′-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene (in the above formula (5), m is 2, and hydroxyl groups are 3- and 4-positions) A compound having a group, hereinafter referred to as a boric acid group-containing trans-stilbene compound (B2).) 0.20 g (0.71 mmol) was obtained. At this time, the yield was 83%. The physical properties of the boric acid group-containing trans-stilbene compound (B2) are shown below. The boric acid group-containing trans-stilbene compound (B2) was obtained by converting one or two hydroxyl groups of the 3′-position dihydroxyboryl group into a methoxy group in a methanol solvent. It was detected with 4′-trihydroxyl-3 ′-(dimethoxyboryl) trans-stilbene and 3,4,4′-trihydroxyl-3 ′-(hydroxyl-methoxyboryl) trans-stilbene.
· MS (ESI negative) m / z; 299 (C 14 H 10 O 3 B (OMe) 2), 285 (C 14 H 10 O 3 B (OMe) OH), 271 (MH)

(Evaluation of physiological activity of boric acid group-containing trans-stilbene compound)
(I) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (A) on B-16 cells Instead of boric acid group-containing trans-stilbene compound (A1), boric acid group-containing trans-stilbene compound (A1) The cell growth inhibitory concentration (IC50) of the boric acid group-containing trans-stilbene compound (A2) with respect to B-16 cells was 6.0 × 10 6 except that A2) was used. It was ^-5 mol / L.

(Ii) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (B) against B-16 cells Instead of boric acid group-containing trans-stilbene compound (A1), boric acid group-containing trans-stilbene compound (A1) The cell growth inhibitory concentration (IC50) of the boric acid group-containing trans-stilbene compound (B2) with respect to B-16 cells was 4.8 × 10 4 except that B2) was used. It was ^-5 mol / L.

(Coupling step (B))
Instead of 3,5-dimethoxybenzyl phosphoric acid diethyl ester, 2,5-dimethoxybenzyl phosphoric acid diethyl ester (in the general formula (6), n is 2, Y is phosphoric acid diethyl ester group (-PO ( OEt) ₂ ), a compound having a methoxy group at the 2-position and 5-position)) except that 1.44 g (5.0 mmol) is used, and the same procedure as in Example 6 is carried out to obtain 2,4 ′, 5 0.91 g (2.6 mmol) of -trimethoxy-3′-bromo-trans-stilbene (hereinafter referred to as a leaving group-containing trans-stilbene compound (F3)) was obtained in a yield of 52%. The physical properties of the leaving group-containing trans-stilbene compound (F3) are shown below.

(Boric acid group introduction process)
The same procedure as in Example 6 was performed except that 0.35 g (1.0 mmol) of the leaving group-containing trans-stilbene compound (F3) was used instead of the leaving group-containing trans-stilbene compound (F1). As a result, 2,4 ′, 5-trimethoxy-3 ′-(dihydroxylboryl) trans-stilbene (in the above formula (4), n is 2 and methoxy at 2- and 5-positions) A compound having a group, hereinafter referred to as a boric acid group-containing trans-stilbene compound (A3).) 0.21 g (0.67 mmol) was obtained. At this time, the yield was 67%. The physical properties of the boric acid-containing trans-stilbene compound (A3) are shown below.

(Hydroxylation step)
The same procedure as in Example 6 was performed except that 0.31 g (1.0 mmol) of the boric acid group-containing trans-stilbene compound (A3) was used instead of the boric acid group-containing trans-stilbene compound (A1). As a result, 2,4 ′, 5-trihydroxyl-3 ′-(dihydroxylboryl) trans-stilbene of brown solid (in the above formula (5), m is 2, and hydroxyl groups are 2- and 5-positions). A compound having a group, hereinafter referred to as a boric acid group-containing trans-stilbene compound (B3).) 0.17 g (0.62 mmol) was obtained. At this time, the yield was 62%. The physical properties of the boric acid group-containing trans-stilbene compound (B3) are shown below. The boric acid group-containing trans-stilbene compound (B3) had a hydroxyl group of the dihydroxyboryl group at the 3′-position converted to a methoxy group, 2,4 ′, 5-trihydroxyl-3 ′-( Dimethoxyboryl) trans-stilbene was detected.
・ MS (ESI negative) m / z; 299 (C ₁₄ H ₁₀ O ₃ B (OMe) ₂ )

(Evaluation of physiological activity of boric acid group-containing trans-stilbene compound)
(I) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (A) on B-16 cells Instead of boric acid group-containing trans-stilbene compound (A1), boric acid group-containing trans-stilbene compound (A1) A cell growth inhibitory concentration (IC50) of the boric acid group-containing trans-stilbene compound (A3) for B-16 cells was 6.0 × 10 6 except that A3) was used. It was ⁻⁶ mol / L.

(Ii) Evaluation of cell growth inhibitory activity of boric acid group-containing trans-stilbene compound (B) against B-16 cells Instead of boric acid group-containing trans-stilbene compound (A1), boric acid group-containing trans-stilbene compound (A1) Except for B3), the cell growth inhibitory concentration (IC50) of the boric acid group-containing trans-stilbene compound (B3) for B-16 cells was 6.0 × 10. It was ⁻⁶ mol / L.

  The boric acid group-containing cis-stilbene compound (1) of the present invention is a pharmaceutical product having a tubulin polymerization inhibitory effect and is a pharmaceutical product whose physiological activity is difficult to decrease in vivo, or a pharmaceutical product whose behavioral analysis in vivo is easy. It is suitably used for production. The boric acid group-containing trans-stilbene compound (A) of the present invention and the boric acid group-containing trans-stilbene compound (B) of the present invention are drugs having an apoptosis-inducing action, and their physiological activities are unlikely to decrease in vivo. It is suitably used for the manufacture of pharmaceuticals or pharmaceuticals that can be easily analyzed in vivo.

Claims (6)

The following general formula (1):

(In the formula, R ¹ represents a hydrogen atom or a methoxy group, R ² represents a hydrogen atom, a boric acid group or a methoxy group, R ³ represents a hydrogen atom, a boric acid group or a methoxy group, and R ¹ , R ² And R ³ may be the same or different, and either R ² or R ³ is a boric acid group.) A boric acid group-containing cis-stilbene compound. The following general formula (2):

(Wherein R ¹ represents a hydrogen atom or a methoxy group) and the following general formula (3):

(Wherein X represents a leaving group, R ⁵ represents a hydrogen atom, a protected boric acid group or a methoxy group, R ⁶ represents a hydrogen atom, a protected boric acid group or a methoxy group, and R ⁵ or R ⁶ Any one is a protected boric acid group.) A coupling step (A) for obtaining a protected boric acid group-containing disubstituted alkyne compound by reacting a protected boric acid group-containing benzene compound represented by Hydrogenation of the protected boric acid group-containing disubstituted alkyne compound obtained in A) to obtain a protected boric acid group-containing cis-stilbene compound, and the protected boric acid group-containing cis obtained in the hydrogenation step -A method for producing a boric acid group-containing cis-stilbene compound, comprising a deprotection step of removing a boric acid group-protecting group of the stilbene compound to obtain a boric acid group-containing cis-stilbene compound. The following general formula (4):

(Wherein R ⁷ represents a methoxy group and n is 2 or 3). A boric acid group-containing trans-stilbene compound (A) represented by: The following general formula (5):

(Wherein R ⁸ represents a hydroxyl group, and m is 2 or 3). A boric acid group-containing trans-stilbene compound (B) represented by: The following general formula (6):

(Wherein R ⁷ represents a methoxy group, Y represents a halogenated phosphonium group, a phosphoric acid group or a phosphoric acid ester group, and n is 2 or 3) and the following general formula ( 7):

(Wherein Z represents a leaving group). A coupling step (B) for obtaining a leaving group-containing trans-stilbene compound by reacting with a benzaldehyde derivative represented by the following formula: obtained in the coupling step (B). A boric acid group having a boric acid group introduction step of substituting the leaving group of the leaving group-containing trans-stilbene compound with a boric acid group to obtain a boric acid group-containing trans-stilbene compound (A) A process for producing a containing trans-stilbene compound (A). The following general formula (6):

(Wherein R ⁷ represents a methoxy group, Y represents a halogenated phosphonium group, a phosphoric acid group or a phosphoric acid ester group, and n is 2 or 3) and the following general formula ( 7):

(Wherein Z represents a leaving group). A coupling step (B) for obtaining a leaving group-containing trans-stilbene compound by reacting with a benzaldehyde derivative represented by the following formula: obtained in the coupling step (B). The leaving group of the leaving group-containing trans-stilbene compound is substituted with a boric acid group to obtain a boric acid group-containing trans-stilbene compound (A). Boric acid characterized by having a hydroxylation step of converting a methoxy group of the boric acid group-containing trans-stilbene compound (A) to a hydroxyl group to obtain a boric acid group-containing trans-stilbene compound (B) A method for producing a group-containing trans-stilbene compound (B).