JP3163391B2 - Stilbene derivatives and anticancer agents containing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to cis-stilbene derivatives and anticancer drugs containing these compounds as active ingredients.

[0002]

BACKGROUND OF THE INVENTION Combretastatins having cis-stilbene as a basic skeleton are known to exhibit strong mitotic inhibitory activity and have strong cytotoxicity, but the compounds are hardly soluble in water. For reasons, it has not been put to practical use as a pharmaceutical. For this reason, various studies have been made on the development of derivatives thereof (CHII M LIN et al, Molecular Phar
macology 34, 200-206, 1988. Mark Cushman et al, J.
Med. Chem., 1991, 34, 2579-2588, International Patent Publication WO92 / 16486, Mark Cushman et al, J. Med. C
hem., 1992, 35, 2293-2306, International Patent Publication WO
93/23357, Mark Cushman et al, J. Med. Che.
m., 1993, 36, 2817-2821, Ryuichi Shiraiet al, Bioo
rg. Med. Chem. Let., Vol. 4, No. 5, pp699-704, 199
4) No effective compound has been found yet.

[0003]

DISCLOSURE OF THE INVENTION An object of the present invention is to find a combretastatin derivative which is easy to synthesize, has low toxicity and has a high therapeutic effect, and provides it as an anticancer agent.

[0004]

Means for Solving the Problems The present inventors have conducted an extensive search for compounds having an anticancer activity by binding an amino acid acyl group to various stilbene derivatives, and as a result, a compound represented by the following general formula (1) was obtained. It has been found that it has a remarkable anticancer effect and low toxicity in animal tests, and has completed the present invention. These compounds have not been synthesized so far and are completely novel cis stilbene derivatives.

[0005]

Embedded image (In the formula, X represents a hydrogen atom or a nitrile group, and Y represents an amino acid acyl group.)

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION In the general formula (1), an amino acid acyl group is an acyl group derived from an amino acid, and the amino acids include α-amino acids, β-amino acids, and γ
-Amino acids. Preferred amino acids are glycine, alanine, leucine, serine, lysine, glutamic acid, aspartic acid, threonine, valine, isoleucine, ornithine, glutamine, asparagine, tyrosine, phenylalanine, cysteine, methionine, arginine, β-alanine, tryptophan, proline,
Histidine and the like are preferred, and threonine or serine is particularly preferred from the viewpoint of drug efficacy and safety. These amino acids are L
Both the isomer and the D-form may be used, but the L-form is preferred.

[0007] Preferred compounds include the following compounds. (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Glycinamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-alanineamide, (Z) -1-
(3-amino-4-methoxyphenyl) -2- (3,
4,5-trimethoxyphenyl) -ethene-β-alanineamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl)
-Ethene-L-leucinamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-serinamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
L-threonine amide, (Z) -1- (3-amino-4
-Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-valinamide, (Z) -1
-(3-Amino-4-methoxyphenyl) -2- (3,
4,5-trimethoxyphenyl) -ethene-L-isoleucinamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L -Prolinamide, (Z) -1- (3
-Amino-4-methoxyphenyl) -2- (3,4,5
-Trimethoxyphenyl) -ethene-L-methioninamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-glutamineamide , (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-glutamylamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
L-aspartylamide, (Z) -1- (3-amino-
4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-asparaginamide.

(Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-lysine amide, (Z) -1- (3-
Amino-4-methoxyphenyl) -2- (3,4,5-
Trimethoxyphenyl) -ethene-L-histidineamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-argininamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-cysteinamide;
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
L-tryptophanamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-thylocyanamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
L-phenylalaninamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-L-ornithinamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
D-alanine amide, (Z) -1- (3-amino-4-
Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-leucinamide, (Z) -1
-(3-Amino-4-methoxyphenyl) -2- (3,
4,5-trimethoxyphenyl) -ethene-D-serinamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl)-
Ethene-D-threonamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-valinamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
D-isoleucinamide, (Z) -1- (3-amino-
4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-prolinamide, (Z)
-1- (3-amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -ethene-D-
Methioninamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-glutamine amide, (Z) -1
-(3-Amino-4-methoxyphenyl) -2- (3,
4,5-Trimethoxyphenyl) -ethene-D-glutamylamide.

(Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-aspartylamide, (Z) -1-
(3-amino-4-methoxyphenyl) -2- (3,
4,5-trimethoxyphenyl) -ethene-D-asparaginamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D -Lysine amide, (Z) -1- (3-
Amino-4-methoxyphenyl) -2- (3,4,5-
(Trimethoxyphenyl) -ethene-D-histidine amide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-argininamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-cysteinamide;
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
D-tryptophanamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-tyrosinamide,
(Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
D-phenylalaninamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -ethene-D-ornithinamide,
(E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-glycinamide, (E) -3- (3
-Amino-4-methoxyphenyl) -2- (3,4,5
-Trimethoxyphenyl) -prop-2-ennitrile-L-alaninamide, (E) -3- (3-amino-4
-Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-β-alaninamide, (E) -3- (3-amino-4-methoxyphenyl) -2- ( 3,4,5-trimethoxyphenyl)
-Prop-2-ennitrile-L-leucinamide,
(E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-L-isoleucinamide, (E)-
3- (3-amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-L-serinamide, (E) -3- (3-
Amino-4-methoxyphenyl) -2- (3,4,5-
Trimethoxyphenyl) -prop-2-ennitrile-
L-threoninamide, (E) -3- (3-amino-4
-Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-phenylalanine amide.

(E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-tyrosinamide, (E) -3 -(3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-prolinamide, (E)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-L-lysine amide, (E) -3- (3-
Amino-4-methoxyphenyl) -2- (3,4,5-
Trimethoxyphenyl) -prop-2-ennitrile-
L-histidine amide, (E) -3- (3-amino-4
-Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-argininamide, (E) -3- (3-amino-4-methoxyphenyl) -2- ( 3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-cysteinamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxy Phenyl) -prop-2-ennitrile-L-methioninamide,
(E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-L-tryptophanamide, (E)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-L-α-aspartylamide, (E)-
3- (3-amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-L-β-aspartylamide, (E)-
3- (3-amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-L-asparaginamide, (E) -3-
(3-amino-4-methoxyphenyl) -2- (3,
4,5-Trimethoxyphenyl) -prop-2-ennitrile-L-α-glutamylamide, (E) -3- (3
-Amino-4-methoxyphenyl) -2- (3,4,5
-Trimethoxyphenyl) -prop-2-ennitrile-L-γ-glutamylamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl)- Prop-2-ennitrile-L-
Glutamide, (Z) -1- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-L-ornithinamide.

(E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-alaninamide, (E) -3 -(3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-leucinamide, (E)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-D-isoleucinamide, (E) -3-
(3-amino-4-methoxyphenyl) -2- (3,
4,5-trimethoxyphenyl) -prop-2-ennitrile-D-serinamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl)- Prop-2-ennitrile-D-threonamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D- Phenylalanine amide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-tyrosinamide, (E) -3 -(3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-prolinamide, (E)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-D-lysine amide, (E) -3- (3-
Amino-4-methoxyphenyl) -2- (3,4,5-
Trimethoxyphenyl) -prop-2-ennitrile-
D-histidine amide, (E) -3- (3-amino-4
-Methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-argininamide, (E) -3- (3-amino-4-methoxyphenyl) -2- ( 3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-cysteinamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxy Phenyl) -prop-2-ennitrile-D-methioninamide,
(E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-D-tryptophanamide.

(E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D-α-aspartylamide, (E ) -3- (3-Amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl)
-Prop-2-ennitrile-D-β-aspartylamide, (E) -3- (3-amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2- Ennitrile-D-asparaginamide,
(E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-D-α-glutamylamide, (E)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
Ennitrile-D-γ-glutamylamide, (E) -3
-(3-Amino-4-methoxyphenyl) -2- (3,
4,5-Trimethoxyphenyl) -prop-2-ennitrile-D-glutamine amide. (Z) -1- (3-Amino-4-methoxyphenyl) -2- (3,4,5-trimethoxyphenyl) -prop-2-ennitrile-D
-Ornithinamide.

The compound of the present invention represented by the general formula (1) can be synthesized, for example, according to the following production route.

[0014]

Embedded image (In the formula, Fmoc represents an N-α-9-fluorenylmethoxycarbonyl group, and AA represents an amino acid acyl group.)

[0015]

Embedded image (Wherein, Fmoc and AA have the same meanings as described above, and Boc
Represents a t-butoxycarbonyl group. )

The compound represented by the formula (5) belonging to the compound of the present invention is, for example, a compound represented by the formula (2): (Z) -1-
(3-amino-4-methoxyphenyl) -2- (3,
4,5-Trimethoxyphenyl) -ethene and an N-α-9-fluorenylmethoxycarbonyl amino acid derivative represented by the formula (3) are added at room temperature in dimethylformamide to dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzoic acid. 6-1 in the presence of triazole (HOBt)
After reacting for 2 hours, purification is performed using chromatography or the like to obtain an intermediate (4), which can be obtained by deprotecting the intermediate with aqueous sodium hydroxide.

The compound represented by the formula (10) belonging to the compound of the present invention is, for example, (E) represented by the formula (6)
-3- (3-Amino-4-methoxyphenyl) -2-
(3,4,5-trimethoxyphenyl) -prop-2-
An ennitrile and an N-α-t-butoxycarbonyl amino acid derivative represented by the formula (7) are combined with 1-ethyl-3- (3-dimethylaminopropyl) in dimethylformamide.
The compound (9) is synthesized by reacting at 50 ° C. for 4 hours in the presence of carbodiimide (WSCI) and then deprotected with hydrochloric acid-dioxane. Alternatively, a compound represented by the formula (6) and an amino acid derivative represented by the formula (3) are mixed with benzotriazol-1-yl-oxy-in acetonitrile.
Synthesis of (8) by reacting at 60 ° C. for 24 hours in the presence of tris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent) and triethylamine, followed by deprotection with aqueous sodium hydroxide or piperidine Can be.

The stilbene derivative of the present invention produced by the above method can be easily separated and purified from the reaction mixture by a conventional isolation and purification means, for example, extraction with a solvent, chromatography, crystallization and the like. .

In the present invention, when the stilbene derivative is used as an anticancer drug, it is administered orally or parenterally (intramuscularly, subcutaneously, intravenously, suppository, etc.). The dose varies depending on the symptoms, but is usually 1 to 9000 mg per day in several divided doses, generally in the range of 1 to 3000 mg per adult.

When an oral preparation of the stilbene derivative of the present invention is prepared, an excipient and, if necessary, a binder, a disintegrant, a lubricant, a coloring agent, a flavoring agent and the like are added. Into tablets, coated tablets, granules, capsules and the like in a conventional manner. As an excipient, for example, lactose, corn starch, sucrose, sugar, sorbitol, crystalline cellulose, etc., as a binder, for example, polyvinyl alcohol, polyvinyl ether, ethyl cellulose, methyl cellulose, gum arabic, tragacanth, gelatin, shellac, hydroxypropyl Examples include cellulose, hydroxypropyl starch, and polyvinylpyrrolidone.

Disintegrators include, for example, starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium bicarbonate, calcium citrate, dextran, pectin and the like. Lubricants include, for example, magnesium stearate, talc, polyethylene glycol. , Silica, hydrogenated vegetable oils, etc. are permitted to be added to pharmaceuticals as coloring agents, but as flavoring agents, cocoa powder, peppermint brain, aromatic acid, peppermint oil, dragon brain, cinnamon powder and the like are used. Can be In these tablets, the granules may be sugar-coated, gelatin-coated, or appropriately coated as needed.

When preparing an injection, the pH may be adjusted as necessary.
Adjustments, buffers, stabilizers, preservatives, and the like are added, and subcutaneous, intramuscular, and intravenous injections are prepared in a conventional manner. The stilbene derivative of the present invention may be, if necessary, pharmaceutically acceptable with an inorganic salt such as hydrochloric acid, sulfuric acid or phosphoric acid, and an organic salt such as oxalic acid, fumaric acid, maleic acid, malic acid, citric acid, tartaric acid or glutamic acid. And acid addition salts which can be used.

[0023]

EXAMPLES The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

Example 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of glycinamide

Step 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of Fmoc-glycinamide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene 2.0 g (6.3 mmol), Fmoc-Gly2.3
g, BOP reagent 11 g (25 mmol) was dissolved in dimethylformamide 40 ml and heated at 60 ° C. for 2 hours.
After cooling, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted three times with dichloromethane. Dried over anhydrous sodium sulfate,
The residue was dried under reduced pressure. Purification by silica gel column chromatography (ethyl acetate: hexane / 1: 2) gave 1.63 g of the desired product. (Yield 43.5%)

1H-NMR (CDCl3) δ; 8.29 (1H, s), 8.11 (1H
, s), 7.76 (2H, d, J = 7.5), 7.60 (2H, d, J = 7.5), 7.39
(2H, t, J = 7.2), 7.30 (2H, m), 7.00 (1H, dd, J = 1.8, 8.
7), 6.70 (1H, d, J = 8.7), 6.51 (1H, d, J = 12, 3), 6.44
(1H, d, J = 12.3), 4.44 (2H, d, J = 6.6), 4.25 (1H, m),
4.04 (2H, 2H, br), 3.84 (3H, s), 3.79 (3H, s), 3.68 (6
H, s); Mass spectrum m / z: 594 (M ⁺ )

Step 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of glycinamide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
1.08 g of Fmoc-glycinamide (1.82 mmol
1) was dissolved in methanol (20 ml), 2N aqueous sodium hydroxide (1.0 ml, 2.0 mmol) was added, and the mixture was stirred for 3 hours. Saturated aqueous sodium bicarbonate was added, extracted three times with dichloromethane, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Purification on a silica gel plate (5% methanol / dichloromethane) gave 479 mg of the desired product. (70.7% yield)

1H-NMR (CDCl3) δ; 9.61 (1H, brs), 8.36 (1
H, d, J = 1.8), 7.00 (1H, dd, J = 1.8,8.4), 6.72 (1H, d,
J = 8.4), 6.51 (2H, s), 6.53 (1H, d, J = 12.0), 6.42 (1
H, d, J = 12.0), 3.87 (3H, s), 3.83 (3H, s), 3.68 (6H,
s); Mass spectrum m / z: 373 (MH ⁺ ); High-resolution mass spectrum Calculated 373.1763, Measured value 373.1751

Example 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-alanine amide

Step 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of Fmoc-L-alanine amide (Z) -1- (3-amino-4-methoxyphenyl)-
2.2 g (6.9 mmol) of 2- (3,4,5-trimethoxyphenyl) -ethene, Fmoc-L-Ala
2.7 g (8.3 mmol), BOP reagent 12.1 g
(27.6 mmol) in 22 ml of dimethylformamide
And heated at 60 ° C. for 4 hours. After cooling, saturated aqueous sodium hydrogen carbonate was added, and the mixture was extracted three times with dichloromethane. It was dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. Purification by silica gel column chromatography (ethyl acetate: hexane / 1: 2) gave 1.79 g of the desired product.
(Yield 41.4%)

1H-NMR (CDCl3) δ; 8.32 (1H, d, J = 1.8),
8.19 (1H, brs), 7.76 (2H, d, J = 7.2), 7.59 (2H, d, J = 7.
2), 7.39 (2H, t, J = 6.9), 7.32 (2H, m), 7.01 (1H, dd, J
= 1.8,8.7), 6.69 (1H, d, J = 8.4), 6.52 (2H, s), 6.51 (1
H, d, J = 12.0), 6.44 (1H, d, J = 12.0), 5.35 (1H, brs),
4.42 (3H, br), 4.24 (1H, m), 3.84 (3H, s), 3.79 (3H,
s), 3.69 (6H, s), 1.48 (3H, d, J = 6.9); mass spectrum m / z: 608 (M ⁺ )

Step 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-alanine amide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
1.0 g of Fmoc-L-alaninamide (1.6 mmo
l) was dissolved in 10 ml of methanol, and 0.9 ml (1.76 mmol) of 2N aqueous sodium hydroxide was added thereto, followed by stirring for 3 hours. Saturated saline was added, extracted with dichloromethane three times, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Purification by silica gel plate (5% methanol / dichloromethane) gave 543 mg of the desired product. (87.9% yield)

1H-NMR (CDCl3) δ; 9.72 (1H, brs), 8.39 (1
H, d, J = 2.1), 6.99 (1H, dd, J = 2.1,8.4), 6.71 (1H, d,
J = 8.4), 6.52 (1H, d, J = 12.3), 6.52 (2H, s), 6.42 (1
H, d, J = 12.3), 3.86 (3H, s), 3.83 (3H, s), 3.68 (6H,
s), 3.64 (1H, m), 1.43 (3H, d, J = 7.2); Mass spectrum
m / z: 387 (MH ⁺ ); high-resolution mass spectrum calculated 38
7.1920, measured value 387.1922

Example 3 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-leucinamide

Step 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of Fmoc-L-leucinamide (Z) -1- (3-amino-4-methoxyphenyl)-
1.92 g (6.1 mmol) of 2- (3,4,5-trimethoxyphenyl) -ethene, Fmoc-L-Leu
2.58 g (7.3 mmol), DCC 1.5 g (7.
3 mmol), HOBt.H ₂ O 1.1 g (7.3 mm
ol) was dissolved in 40 ml of dimethylformamide and reacted at room temperature for 12 hours. After dilution with ethyl acetate, the mixture was filtered and concentrated. Purification by silica gel column chromatography (ethyl acetate: hexane / 1: 2) gave the desired product 3.05.
g was obtained. (76.9% yield)

1H-NMR (CDCl3) δ; 8.32 (1H, d, J = 2.1),
8.19 (1H, s), 7.75 (2H, d, J = 7.5), 7.58 (2H, d, J = 7.
5), 7.39 (2H, t, J = 6.9), 7.29 (2H, m), 7.00 (1H, dd, J
= 2.1, 8.4), 6.69 (1H, d, J = 8.4), 6.51 (2H, s), 6.50
(1H, d, J = 12.3), 6.43 (1H, d, J = 12.3), 5.29 (1H, br
s), 4.43 (2H, d, J = 6.9), 4.23 (1H, t, J = 6.9), 3.83 (3
H, s), 3.79 (3H, s), 3.68 (6H, s), 1.75 (2H, br), 11.
55 (1H, br), 0.95 (6H, br); mass spectrum m / z: 650 (
M ⁺ )

Step 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-leucinamide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
1.0 g of Fmoc-L-leucinamide (1.54 mm
ol) in 10 ml of methanol and 10 ml of dichloromethane.
0.9N 2N sodium hydroxide solution (1.7m
mol) was added and stirred for 3 hours. Saturated saline was added, extracted with dichloromethane three times, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Purification by a silica gel column (10% methanol / dichloromethane) gave 560 mg of the desired product. (84.9% yield)

1H-NMR (CDCl3) δ; 9.78 (1H, brs), 8.41 (1
H, d, J = 1.8), 6.99 (1H, dd, J = 1.81,8.4), 6.70 (1H,
d, J = 8.4), 6.52 (1H, d, J = 12.3), 6.52 (2H, s), 6.42
(1H, d, J = 8.4), 3.87 (3H, s), 3.83 (3H, s), 3.68 (6H,
s), 3.51 (1H, m), 1.80 (2H, m), 1.42 (1H, m), 0.98 (6
(H, t, J = 6.6) Mass spectrum m / z: 429 (MH ⁺ ); High resolution mass spectrum Calculated 429.2389, Measured 429.2
391

Example 4 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-serinamide

Step 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of Fmoc-L-serinamide (Z) -1- (3-amino-4-methoxyphenyl)-
1.5 g (4.76 mmol) of 2- (3,4,5-trimethoxyphenyl) -ethene, Fmoc-L-Ser
(Ac) 2.1 g (5.7 mmol), DCC 1.2 g
(5.7 mmol), 0.87 g of HOBt.H ₂ O
(5.7 mmol) was dissolved in 30 ml of dimethylformamide and reacted at room temperature for 5 hours. After dilution with ethyl acetate, the mixture was filtered and concentrated. Purification by silica gel column chromatography (ethyl acetate: hexane / 1: 2) gave 1.96 g of the desired product. (Yield 61.8%)

1H-NMR (CDCl ₃ ) δ; 8.38 (1H, br), 8.30 (1H,
d, J = 1.8), 7.76 (2H, d, J = 7.8), 7.59 (2H, d, J = 7.
8), 7.40 (2H, t, J = 7.2), 7.32 (2H, m), 7.03 (1H, dd, J
= 1.8, 8.7), 6.71 (1H, d, J = 8.7), 6.51 (2H, s), 6.51
(1H, d, J = 12.3), 6.45 (1H, d, J = 12.3), 5.53 (1H, br
s), 4.62 (1H, br), 4.45 (2H, d, J = 6.9), 4.25 (1H, m),
3.83 (3H, s), 3.80 (3H, s), 3.69 (6H, s), 2.65 (2H,
d, J = 9.3), 2.1 (3H, s); mass spectrum m / z: 666 (M
⁺ )

Step 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-serinamide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
1.04 g of Fmoc-L-serinamide (1.56 mm
ol) in 10 ml of methanol and 10 ml of dichloromethane.
1.7 ml of 2N aqueous sodium hydroxide (3.4 m
mol) and stirred at room temperature for 24 hours. Saturated saline was added, extracted with dichloromethane three times, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Silica gel plate (5
% Methanol / dichloromethane).
5 mg were obtained. (Yield 50.2%)

1H-NMR (CDCl3) δ; 9.77 (1H, brs), 8.34 (1
H, d, J = 2.1), 7.01 (1H, dd, J = 2.1,8.7), 6.73 (1H, d,
J = 8.7), 6.52 (2H, s), 6.51 (1H, d, J = 12.3), 6.43 (1
H, d, J = 12.3), 3.98 (1H, dd, J = 4.8, 11.1), 3.87 (3H,
s), 3.84 (3H, s), 3.79 (1H, dd, J = 5.4, 11.1), 3.69 (6
H, s), 3.59 (1H, dd, J = 5.1, 5.4); mass spectrum m /
z: 403 (MH +); High-resolution mass spectrum calculated value 403.18
69, measured value 403.1862

Example 5 (Z) -1- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-threonine amide

Step 1 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of Fmoc-L-threonine (Ac) amide (Z) -1- (3-amino-4-methoxyphenyl)-
1.5 g (4.76 mmol) of 2- (3,4,5-trimethoxyphenyl) -ethene, Fmoc-L-Thr
(Ac) 2.2 g (5.7 mmol), DCC 1.2 g
(5.7 mmol), 0.87 g of HOBt.H ₂ O
(5.7 mmol) was dissolved in 30 ml of dimethylformamide and reacted at room temperature for 6 hours. 50 ml of ethyl acetate
, And concentrated by filtration. Purification by silica gel column chromatography (ethyl acetate: hexane / 1: 2) gave 2.97 g of the desired product. (Yield 91%)

1H-NMR (CDCl3) δ; 8.36 (1H, brs), 8.29 (1
H, d, J = 2.4), 7.77 (2H, m), 7.61 (2H, m), 7.28-7.44
(4H, m), 7.02 (1H, dd, J = 2.1, 8.7), 6.72 (1H, d, J =
8.7), 6.51 (2H, s), 6.51 (1H, d, J = 12.0), 6.45 (1H,
d, J = 12.0), 5.72 (1H, m), 5.40 (1H, m), 4.48 (2H, m),
4.25 (1H, m), 3.83 (3H, s), 3.82 (3H, s), 3.69 (6H,
s), 2.08 (3H, s), 1.24 (3H, m); mass spectrum m / z: 6
80 (M ⁺ )

Step 2 (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
Synthesis of L-threonine amide (Z) -1- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -ethene-
1.0 g of Fmoc-L-threonine (Ac) amide
(1.47 mmol) in 20 ml of dioxane
1.76 ml of N sodium hydroxide aqueous solution (3.5 mmol)
Was added and stirred for 24 hours. Saturated saline was added, extracted with dichloromethane three times, dried over anhydrous sodium sulfate,
The residue was dried under reduced pressure. Silica gel plate (7.5% methano-
(Dichloromethane / dichloromethane) to obtain 448 mg of the desired product. (Yield 73.4%)

1H-NMR (CDCl3) δ; 9.86 (1H, brs), 8.37 (1
H, d, J = 2.1), 7.01 (1H, dd, J = 2.1,8.7), 6.72 (1H, d,
J = 8.7), 6.52 (2H, s), 6.52 (1H, d, J = 12.0), 6.43 (1
H, d, J = 12.0), 4.42 (1H, m), 3.87 (3H, s), 3.84 (3H,
s), 3.69 (6H, s), 3.38 (1H, m), 1.25 (3H, d, J = 6.3);
Mass spectrum m / z: 417 (MH ⁺ ); High resolution mass spectrum Calculated 417.2026, Measured 417.2050

Example 6 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-glycinamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-glycinamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
700 mg of 2-ennitrile hydrochloride (1.86 mmol
l), WSCI 478 mg, HOBt.H ₂ O 375 m
g, Boc-Gly486mg were dissolved in dimethylformamide 100ml, triethylamine 0.35ml was added, and reacted at 50 ° C for 3.5 hours. After adding 700 ml of water and extracting with ethyl acetate, the ethyl acetate layer was washed three times with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification on a silica gel column (developing solvent ether),
Dissolved in a small amount of dichloromethane, and crystallized by adding ether to obtain 851 mg of the desired product (1.71 mmol,
92%).

1H-NMR (CDCl3) δ; 1.481 (s, 9H), 3.759 (s,
6H), 3.855 (s, 3H), 3.883 (s, 3H), 3.901 (d, J = 5.7Hz),
5.1 (br, 1H), 6.603 (s, 2H), 6.696 (d, J = 8.5Hz, 1H), 6.89
2 (dd, J = 1.8Hz, 8.5Hz, 1H), 7.245 (s, 1H), 8.295 (br.s, 1
H), 8.333 (d, J = 1.8 Hz, 1H); Mass spectrum: 497 (M ⁺ ).

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-glycinamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
800 mg of 2-ennitrile-Boc-glycinamide
Was dissolved in 3 ml of dichloromethane, 3 ml of a 4 M hydrochloric acid-dioxane solution was added, and the mixture was reacted at room temperature for 2 hours. 30 ml of ether was added and the mixture was filtered.
It was washed with isopropanol-toluene (6: 8: 20) while hot, and 483 mg (1.11 mmol, 65%) of the desired product was obtained.
I got

1H-NMR (CD3OD) δ; 3.735 (s, 6H), 3.807 (b
r, 2H), 3.812 (s, 3H), 3.888 (s, 3H), 6.662 (s, 2H), 6.97
8 (d, J = 8.6Hz, 1H), 7.102 (dd, J = 2.1Hz, 8.6Hz, 1H), 7.3
46 (s, 1H), 8.018 (d, J = 2.1Hz, 1H) ； High-resolution mass spectrum calculated 398.1716, measured 398.1723

Example 7 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-ornithinamide dihydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-ornithinamide (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
700 mg of 2-ennitrile hydrochloride (1.86 mmol
l), WSCI 463 mg, HOBt.H ₂ O 463 m
g, Boc2-L-Orn 767 mg was dissolved in dimethylformamide 70 ml, and triethylamine 0.35 m
1 was added and reacted at 50 ° C. for 41 hours. 400 ml of water
After extraction with ether, the ether layer was washed three times with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure.
After purification with a silica gel column (developing solvent ether), the residue was dissolved in a small amount of dichloromethane and crystallized by adding ether to obtain 737 mg (1.13 mmol, 61%) of the desired product.

1H-NMR (CDCl3) δ; 1.432 (s, 9H), 1.451 (s,
9H), 1.5 (m, 2H), 1.65 (m, 1H), 1.9 (m, 1H), 3.2 (m, 2H),
3.764 (s, 6H), 3.857 (s, 3H), 3.875 (s, 3H), 4.2 (br, 1H),
4.8 (br, 1H), 5.1 (br, 1H), 6.600 (s, 2H), 6.704 (d, J = 8.6
Hz, 1H), 6.901 (dd, J = 2.1Hz, 8.6Hz, 1H), 7.236 (s, 1H),
8.266 (d, J = 2.1Hz, 1H), 8.329 (br.s, 1H); Mass spectrum: 654 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-ornithinamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Boc-L-ornithinamide 73
0 mg (1.11 mmol) was dissolved in 5 ml of dichloromethane, 5 ml of a 4 M hydrochloric acid-dioxane solution was added, and the mixture was reacted at room temperature for 1 hour. 100 ml of ether was added and filtered,
The obtained powder was recrystallized from methanol-ethyl acetate (1: 1) to give 286 mg (0.542 mmol,
8%).

1H-NMR (CDCl3) δ; 1.7 (m, 2H), 1.9 (m, 2H),
2.973 (d, J = 6.3Hz, 1H), 3.003 (d, J = 6.3Hz, 1H), 3.768
(s, 6H), 3.820 (s, 3H), 3.898 (s, 3H), 4.176 (t, J = 6.3Hz,
1H), 6.675 (s, 2H), 7.014 (d, J = 8.5Hz, 1H), 7.173 (dd,
J = 2.0Hz, 8.5Hz, 1H), 7.368 (s, 1H), 7.801 (d, J = 2.0Hz, 1
H); High-resolution mass spectrum; Calculated 455.2288, Measured 455.2300

Example 8 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-phenylalaninamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-phenylalanine amide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
998 mg of 2-ennitrile hydrochloride (2.65 mmol
1), BOP reagent 1290 mg, Boc-L-Phe7
77 mg was dissolved in 50 ml of acetonitrile, and 0.8 ml of triethylamine was added. After adding 100 ml of water, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification with a silica gel column (developing solvent: dichloromethane), the residue was dissolved in a small amount of dichloromethane, and crystallized by adding ether and hexane to give 1082 mg of the desired product (1.
84 mmol, 69%).

1H-NMR (CDCl3) δ; 1.426 (s, 9H), 3.12 (br.
t, 2H), 3.744 (s, 3H), 3.766 (s, 6H), 3.888 (s, 3H), 4.4
(br, 1H), 5.1 (br, 1H), 6.613 (s, 2H), 6.639 (d, J = 8.8Hz,
1H), 6.875 (dd, J = 2.1Hz, 8.8Hz, 1H), 7.18-7.36 (m, 5H),
8.030 (br.s, 1H), 8.345 (d, J = 2.1Hz, 1H); FAB mass spectrum: 587 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-phenylalaninamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1082 mg (1.11 mmol) of 2-ennitrile-Boc-L-phenylalaninamide is dissolved in 10 ml of dichloromethane, and 5 ml of a 4 M hydrochloric acid-dioxane solution is dissolved.
The mixture was reacted at room temperature for 1 hour. 100 ml of ether was added and the mixture was filtered. The obtained powder was chloroform-methanol-
Recrystallization from ethyl acetate (4: 1: 4) gave the desired product 450.
mg (0.859 mmol, 77%).

1H-NMR (CD3OD) δ; 3.106 (d, J = 7.3 Hz, 1H),
3.119 (d, J = 7.3Hz, 1H), 4.312 (t, J = 7.3Hz, 1H), 3.751 (s,
6H), 3.792 (s, 3H), 3.819 (s, 3H), 6.672 (s, 2H), 6.936
(d, J = 8.7Hz, 1H), 7.173 (dd, J = 2.2Hz, 8.7Hz, 1H), 7.2
(m, 2H), 7.3 (m, 3H), 7.339 (s, 1H), 7.878 (d, J = 2.2Hz, 1
H); High-resolution mass spectrum calculated 488.2186, measured 488.2162

Example 9 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-prolinamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-prolinamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
998 mg of 2-ennitrile hydrochloride (2.65 mmol
l), 1300 mg of BOP reagent, Boc-L-Pro6
05 mg was dissolved in 50 ml of acetonitrile, and 0.8 ml of triethylamine was added. After 100 ml of water and a small amount of sodium hydrogen carbonate were added, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification on a silica gel column (developing solvent: dichloromethane), the mixture was concentrated to give the desired product 1
310 mg (2.44 mmol, 92%) were obtained.

1H-NMR (CDCl3) δ; 1.4-1.5 (br, 9H), 1.9 (b
r, 2H), 2.1-2.3 (br, 1H), 2.3-2.5 (br, 1H), 3.3-3.5 (br,
2H), 3.753 (s, 6H), 3.838 (s, 3H), 3.876 (s, 3H), 4.2-4.
5 (br, 1H), 6.609 (s, 2H), 6.677 (d, J = 8.4Hz, 1H), 6.871
(m, 1H), 7.238 (s, 1H), 8.39 (br.s, 1H), 9.2 (br, 1H); FA
B mass spectrum: 537 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-prolinamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Boc-L-prolinamide 125
0 mg (2.33 mmol) in dichloromethane 10 ml
And 5 ml of a 4 M hydrochloric acid-dioxane solution was added thereto, and the mixture was reacted at room temperature for 1 hour. 100 ml of ether was added and the mixture was filtered.
S, water-acetonitrile 70:30) and recrystallization three times with chloroform-ethyl acetate 1:10 to obtain 465 mg (0.980 mmol, 42%) of the desired product.

1H-NMR (CDCl3) δ; 2.0 (m, 3H), 2.4 (m, 1H),
3.4 (m, 2H), 3.745 (s, 6H), 3.805 (s, 3H), 3.895 (s, 3H),
4.45 (m, 1H), 6.660 (s, 2H), 6.997 (d, J = 8.6Hz, 1H), 7.
143 (dd, J = 2.1Hz, 8.6Hz, 1H), 7.349 (s, 1H), 7.839 (d, J
= 2.1Hz, 1H); High-resolution mass spectrum calculated value 438.202
9, measured value 438.2033

Example 10 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-alanine amide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-alanine amide (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1053 mg (2.65 mmol of 2-ennitrile hydrochloride)
l), 1300 mg of BOP reagent, Boc-L-Ala5
54 mg was dissolved in 50 ml of acetonitrile, 0.8 ml of triethylamine was added, and the mixture was reacted at 60 ° C. for 17 hours. Add 100 ml of water and a small amount of sodium bicarbonate,
The mixture was extracted with ethyl acetate, washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification on a silica gel column (developing solvent dichloromethane-ethyl acetate 20:
1) Concentrate and concentrate 1085 mg (2.12 mmol
1, 80%).

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-alanine amide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Boc-L-alaninamide 100
0 mg (1.95 mmol) in 10 ml of dichloromethane
And 5 ml of a 4 M hydrochloric acid-dioxane solution was added thereto, and the mixture was reacted at room temperature for 1 hour. After adding 100 ml of ether and filtering, the obtained powder is recrystallized twice with chloroform-methanol-ethyl acetate 20: 2: 30, and the obtained powder is subjected to medium pressure liquid chromatography (ODS, water-acetonitrile 75:25). The precipitate formed by adding ether to a small amount of methanol was collected by filtration, and 280 mg of the desired product was obtained.
(0.625 mmol, 32%).

1H-NMR (CD3OD) δ; 1.503 (d, J = 7.0 Hz, 3H),
3.736 (s, 6H), 3.808 (s, 3H), 3.888 (s, 3H), 4.129 (q, J =
7.0Hz, 1H), 6.662 (s, 2H), 6.985 (d, J = 8.6Hz, 1H), 7.12
2 (dd, J = 2.3Hz, 8.6Hz, 1H), 7.345 (s, 1H), 7.900 (d, J =
2.3Hz, 1H); High-resolution mass spectrum Calculated value 412.187
3, measured value 412.1873

Example 11 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-threonamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-threonamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1000 mg of 2-ennitrile hydrochloride (2.65 mmol
l), 1300 mg of BOP reagent, Boc-L-Thr
Dissolve 880 mg of (OtBu) in 50 ml of acetonitrile, add 0.8 ml of triethylamine, and add 2 ml at 60 ° C.
The reaction was performed for 1 hour. 100 ml of water and a small amount of sodium bicarbonate were added, extracted with ethyl acetate, washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure.
After purification on a silica gel column (developing solvent: ethyl acetate-hexane 5: 1), the mixture was concentrated, and 870 mg of the desired product (1.46) was obtained.
mmol, 55%).

1H-NMR (CD3OD) δ; 1.044 (d, J = 6.0 Hz, 3H),
1.315 (s, 9H), 1.463 (s, 9H), 3.760 (s, 6H), 3.844 (s, 3
H), 3.887 (s, 3H), 4.15 (br.m, 1H), 4.22 (br, 1H), 5.64
(br.d, 1H), 6.617 (s, 2H), 6.857 (d, J = 8.5Hz, 1H), 6.897
(dd, J = 2.2Hz, 8.5Hz, 1H), 7.228 (s, 1H), 8.404 (d, J = 2.
2Hz, 1H), 9.3 (br.s, 1H); FAB mass spectrum: 597
(M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-threonamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Boc-L-threonamide 81
0 mg (1.95 mmol) in 10 ml of dichloromethane
, And 5 ml of a 4 M hydrochloric acid-dioxane solution was added thereto.
The reaction was carried out at a temperature of 3 ° C. for 3 hours. 100 ml of ether was added and the mixture was filtered. The obtained powder was subjected to medium pressure liquid chromatography (OD
S, purified twice with water-acetonitrile (75:25), dissolved in a small amount of methanol and added with acetonitrile-ethyl acetate, and the resulting precipitate was collected by filtration to give 290 mg (0.60 mg) of the desired product.
7 mmol, 31%).

1H-NMR (CD3OD) δ; 1.240 (d, J = 6.3 Hz, 3H),
3.9 (1H), 3.739 (s, 6H), 3.810 (s, 3H), 3.892 (s, 3H), 4.0
12 (m, 1H), 6.658 (s, 2H), 6.996 (d, J = 8.5Hz, 1H), 7.133
(dd, J = 2.2Hz, 8.5Hz, 1H), 7.350 (s, 1H), 7.923 (d, J = 2.
2Hz, 1H); High-resolution mass spectrum Calculated value 442.1978,
Measured value 442.1973

Example 12 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-lysine amide dihydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Boc-L-lysine amide (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1000 mg of 2-ennitrile hydrochloride (2.65 mmol
l), 1462 mg of Boc2-L-LysOSu was dissolved in 50 ml of acetonitrile, and 0.8 ml of triethylamine was dissolved.
of HOBt60.
0 mg, 1300 mg of BOP reagent and further 60 ° C
For 21 hours. 100 ml of water and a small amount of sodium bicarbonate were added, extracted with ethyl acetate, washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification with a silica gel column (developing solvent: dichloromethane), the mixture was concentrated and 1170 mg (1.74 mmol) of the desired product was obtained.
1, 66%).

1H-NMR (CDCl3) δ; 1.438 (s, 9H), 1.450 (s,
9H), 1.4-1.5 (br, 4H), 1.7 (br, 1H), 1.9 (br, 1H), 3.1 (b
r, 2H), 3.756 (s, 6H), 3.852 (s, 3H), 3.874 (s, 3H), 4.2
(br, 1H), 4.7 (br, 1H), 5.2 (br, 1H), 6.604 (s, 2H), 6.68
5 (d, J = 8.7Hz, 1H), 6.884 (dd, J = 2.2Hz, 8.7Hz, 1H), 7.23
1 (s, 1H), 8.348 (br, 1H); FAB mass spectrum: 668 (
M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-lysine amide dihydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Boc-L-lysine amide 1100
mg (1.64 mmol) was dissolved in 10 ml of dichloromethane, and 5 ml of a 4 M hydrochloric acid-dioxane solution was added thereto.
For 3 hours. 100 ml of ether was added and the mixture was filtered. The obtained powder was subjected to medium pressure liquid chromatography (OD
S, water-acetonitrile 95: 5 → 85: 15), dissolved in a small amount of methanol, acetonitrile-ethyl acetate was added, and the resulting precipitate was collected by filtration.
(0.554 mmol, 34%).

1H-NMR (CD3OD) δ; 1.4 (m, 2H), 1.7 (m, 2H),
1.9 (m, 2H), 2.95 (m, 2H), 3.756 (s, 6H), 3.811 (s, 3H),
3.896 (s, 3H), 4.131 (t, J = 6.3), 6.667 (s, 2H), 7.010 (d,
J = 8.9Hz, 1H), 7.164 (dd, J = 2.3Hz, 8.9Hz, 1H), 7.361 (s,
1H), 7.834 (d, J = 2.3Hz, 1H); High-resolution mass spectrum Calculated 469.2451, Measured 469.2454

Example 13 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-serinamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Fmoc-L-serinamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1007 mg of 2-ennitrile hydrochloride (2.65 mmol
l), Fmoc-L-Ser (OtBu) OH1105
mg, BOP reagent 1370mg, HOBt · H ₂ O6
Dissolve 18 mg in 50 ml of acetonitrile, add 0.8 ml of triethylamine, react at 42 ° C. for 42 hours, add 100 ml of water and a small amount of sodium hydrogen carbonate,
The mixture was extracted with ethyl acetate, washed with saturated saline, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification on a silica gel column (developing solvent: ethyl acetate-hexane 2: 3),
After concentration, 1486 mg (2.14 mmol, 81
%).

1H-NMR (CDCl3) δ; 1.241 (s, 9H), 3.243 (t,
J = 8.5Hz, 1H), 3.760 (s, 6H), 3.832 (s, 3H), 3.874 (s, 3
H), 4.247 (m, 1H), 4.33 (br, 1H), 4.42 (m, 2H), 5.8 (br, 1
H), 6.617 (s, 2H), 6.704 (d, J = 8.8Hz, 1H), 6.904 (dd, J =
2.2Hz, 8.8Hz, 1H), 7.252 (s, 1H), 7.32 (m, 2H), 7.407 (t, J
= 7.5Hz, 2H), 7.612 (d, J = 7.5Hz, 2H), 7.772 (d, J = 7.2Hz, 2
H), 8.406 (d, J = 2.2 Hz), 9.0 (br.s, 1H) FAB; Mass spectrum: 705 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-serinamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-ennitrile-Fmoc-L-serinamide 143
0 mg (2.07 mmol) was dissolved in 5 ml of chloroform and 2 ml of piperidine, reacted for 1 hour, and purified by a silica gel column (developing solvent: ethyl acetate-dichloromethane 1: 1). After concentrating to dryness under reduced pressure, the residue was dissolved in 4 ml of a 4M hydrochloric acid-dioxane solution, reacted at 70 ° C. for 1 hour, and 100 ml of ether was added. The resulting powder was purified by medium pressure liquid chromatography (ODS, water-acetonitrile 75:25), dissolved in chloroform-methanol 5: 1 while hot, ethyl acetate was added, and the resulting precipitate was collected by filtration. 460mg (0.992mm
ol, 48%).

1H-NMR (CD3OD) δ; 3.737 (s, 6H), 3.813 (s,
3H), 3.892 (s, 3H), 3.9 (m, 2H), 4.123 (dd, J = 5.1Hz, 6.3
Hz, 1H), 6.662 (s, 2H), 6.981 (d, J = 8.5Hz, 1H), 7.109 (d-
d, J = 2.2Hz, 8.5Hz, 1H), 7.344 (s, 1H), 7.998 (d, J = 2.2H
z, 1H); High-resolution mass spectrum calculated 428.1822, measured 428.1806

Example 14 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-aspartylamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-Fmoc-L-aspartylamide (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
900 mg (2.65 mmol) of 2-ennitrile, F
1400 mg of moc-L-Asp (OBn), 1300 mg of BOP reagent and 660 mg of HOBt.H ₂ O were dissolved in 50 ml of acetonitrile, and 0.5 ml of triethylamine was dissolved.
was added, and the mixture was reacted at room temperature for 86 hours. 100 ml of water and a small amount of sodium hydrogen carbonate were added, extracted with ethyl acetate, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. After purification on a silica gel column (developing solvent,
Ethyl acetate-dichloromethane 1:10) and concentrated to obtain 1319 mg (1.80 mmol, 68%) of the desired product.

1H-NMR (CDCl3) δ; 2.76 (br.dd, 1H), 3.15
(br.d, 1H), 3.747 (s, 9H), 3.869 (s, 3H), 4.231 (t, J = 7.0
Hz, 1H), 4.457 (m, 2H), 4.72 (br, 1H), 5.133 (d, J = 12.3H
z, 1H), 5.206 (d, J = 12.3Hz, 1H), 6.607 (s, 2H), 6.662 (d, J
= 9.0Hz, 1H), 6.896 (dd, J = 2.1Hz, 9.0Hz, 1H), 7.20-7.45
(m, 4H), 7.342 (s, 1H), 7.58 (br.d, 2H), 7.762 (dd, J = 2.
5Hz, 7.3Hz, 2H), 8.327 (d, J = 2.1Hz), 8.7 (br.s, 1H); FAB
Mass spectrum: 767 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-aspartylamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
1210 mg (1.65 mmol) of 2-ennitrile-Fmoc-L-aspartylamide was added to dioxane 30
Then, 2 ml of 2M aqueous sodium hydroxide was added, and the mixture was reacted at room temperature for 1 hour. Then, 100 ml of ether was added, and the deposited precipitate was collected by filtration. 30 ml of dioxane again
And 0.5 ml of 2M aqueous sodium hydroxide and 1.5 ml of water were added and reacted at room temperature for 1 hour.
0 ml was added, and the deposited precipitate was collected by filtration. The filtrate is purified little by little by medium pressure liquid chromatography (ODS, water-methanol-hydrochloric acid (12N) 75: 25: 0.3). : After dissolving in 200 ml, NaOHaq (2M)
Was added to neutralize the mixture, and the mixture was allowed to stand for 40 minutes, and the deposited precipitate was collected by filtration. The obtained residue was dissolved in 4M hydrochloric acid-dioxane solution 0.3
The precipitate was dissolved in a small amount of methanol including ethyl acetate, and ethyl acetate was added.
94 mmol, 36%).

1H-NMR (CD3OD) δ; 3.08 (m, 2H), 3.752 (s,
6H), 3.812 (s, 3H), 3.868 (s, 3H), 4.256 (t, J = 5.4Hz,
1H), 6.646 (s, 2H), 6.948 (d, J = 8.6Hz, 1H), 7.086 (d
-d, J = 2.0Hz, 8.6Hz, 1H), 7.330 (s, 1H), 7.821 (d, J =
2.0Hz, 1H); High-resolution mass spectrum Calculated value 456.17
71, measured 456.1775

Example 15 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-glutamylamide hydrochloride

Step 1 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-Ennitrile-Fmoc-L-glutamyl (OB
n) Synthesis of amide hydrochloride (E) -3- (3-amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
900 mg (2.65 mmol) of 2-ennitrile, F
moc-L-Glu (OBn) 1500 mg, BOP reagent 1300 mg, HOBt.H ₂ O 643 mg were dissolved in acetonitrile 50 ml, and triethylamine 0.5 m
l, and reacted at room temperature for 64 hours. 100 ml of water and a small amount of sodium bicarbonate were added, and the mixture was extracted with dichloromethane.
After washing with saturated saline, drying over anhydrous sodium sulfate,
It was concentrated under reduced pressure. After purification with a silica gel column (developing solvent: dichloromethane), the mixture was concentrated and 1950 mg of the desired product (2.
55 mmol, 97%).

1H-NMR (CDCl3) δ; 1.9-2.1 (br.m, 1H), 2.
1-2.3 (br.m, 1H), 2.4-2.7 (br.m, 2H), 3.745 (s, 6H),
3.788 (s, 3H), 3.868 (s, 3H), 3.85-3.95 (m, 1H), 4.20
7 (t, J = 6.9Hz, 1H), 4.408 (d, J = 6.9Hz, 2H), 5.137 (s,
2H), 5.6-5.7 (br.s, 1H), 6.603 (s, 2H), 6.675 (d, J =
8.7Hz, 1H), 6.899 (dd, J = 2.0Hz, 8.7Hz, 1H), 7.2-7.
4 (m, 10H), 7.577 (d, J = 7.5Hz, 2H), 7.754 (d, J = 7.5H
z, 2H), 8.320 (m, 2H); FAB mass spectrum: 781 (M ⁺ )

Step 2 (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
Synthesis of 2-ennitrile-L-glutamylamide hydrochloride (E) -3- (3-Amino-4-methoxyphenyl)-
2- (3,4,5-trimethoxyphenyl) -prop-
2-Ennitrile-Fmoc-L-glutamyl (OB
n) 1940 mg (2.55 mmol) of the amide was dissolved in 50 ml of dioxane, and 3.7 ml of 2M aqueous sodium hydroxide was used.
After reacting at room temperature for 1 hour, ether 100 ml
Was added, and the deposited precipitate was collected by filtration. This was again dissolved in 30 ml of dioxane, and 0.5 ml of 2M sodium hydroxide.
After adding 1.5 ml of water and 1.5 hours of reaction at room temperature, 20 ml of methanol was added to the reaction solution, the mixture was poured into 250 ml of ether, and the deposited precipitate was collected by filtration. The residue was purified little by little by medium pressure liquid chromatography (ODS, water-acetonitrile-hydrochloric acid (12N) 75: 25: 0.3), and concentrated so as not to dryness. When the solution became about 50 ml, it was added to ethyl acetate-ether (1: 1) to precipitate. After discarding the supernatant, 110 ml of acetonitrile and 350 ml of ether were sequentially added, and the precipitated precipitate was filtered and eluted. Wash with tel and dry under reduced pressure.
(0.838 mmol, 33%) was obtained.

1H-NMR (CD3OD) δ; 2.120 (q, J = 7.0 Hz, 2
H), 2.468 (m, 2H), 3.735 (s, 6H), 3.808 (s, 3H), 3.88
8 (s, 3H), 4.131 (t, J = 6.3Hz, 1H), 6.658 (s, 2H), 6.9
95 (d, J = 8.6Hz, 1H), 7.143 (dd, J = 2.2Hz, 8.6Hz, 1
H), 7.349 (s, 1H), 7.861 (d, J = 2.2Hz, 1H); high-resolution mass spectrum calculated 470.1927, measured 470.1914

Example 16 Evaluation of Cytotoxicity Mouse P388 leukemia cells were used as cancer cells, and RPMI-1640 medium containing 5 μM 2-mercaptoethanol and 10% fetal bovine serum was used for culture. The cells were seeded at 1 × 10 4 cells / 50 μl / well on a 96-well microplate, and 25% aqueous solution of the test compound solution (4 μg / ml) was added.
After adding each μl and culturing at 37 ° C. for 2 days, the number of viable cells was measured by the MTT method, and a dose-response curve was prepared. From this, 50% growth inhibitory concentration (IC5
0) was calculated. The IC50 values of each compound are shown in the table.
Tables 1 to 4 show doses at which acute deaths occur immediately after drug administration as toxicity expression doses.

Example 17 Drug Efficacy Test in Mice Colon 26 subcutaneously passaged in mice was crushed with scissors and transplanted subcutaneously in mice with a trocar. One week later, the tumor was measured with a caliper and the tumor volume was calculated to perform grouping (n = 3 in each group). After the test compound was dissolved in dimethyl sulfoxide, it was diluted with a 5% Tween 80-physiological saline solution, and 0.2 ml of the solution was administered by intravenous injection.
Drug administration is once a day on days 7, 11, and 15 after transplantation
I went there. On the 21st day after the transplantation, the tumor volume was measured, and the tumor volume and the tumor growth inhibition rate (IR) were calculated by Equations 1 and 2, respectively.

[0100]

(Equation 1)

[0101]

(Equation 2)

[0102]

[Table 1]

[0103]

[Table 2]

[0104]

[Table 3]

[0105]

[Table 4]

[0106]

Industrial Applicability The stilbene derivative of the present invention has excellent anticancer activity and is extremely useful in the pharmaceutical industry.

────────────────────────────────────────────────── ─── Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI C07C 255/44 C07C 255/44 C07D 207/16 C07D 207/16 (72) Inventor Yukio Nihei 1 Suzukicho, Kawasaki-ku, Kawasaki-shi, Kanagawa Prefecture -1 Ajinomoto Co., Ltd. Central Research Laboratory (72) Inventor Ryusuke Nakagawa 1-1 Azinomoto Co., Ltd. Suzukicho, Kawasaki City, Kawasaki City, Kanagawa Prefecture (72) Inventor Kazuo Oishi Suzuki, Kawasaki City, Kawasaki City, Kanagawa Prefecture Machi 1-1 Ajinomoto Co., Inc. Central Research Laboratory (56) References JP-A-7-228558 (JP, A) (58) Field surveyed (Int. Cl. ⁷ , DB name) C07C 237/00-409 / 44 A61K 31/16-31/655 CA (STN) REGISTRY (STN)

Claims (6)

(57) [Claims] 1. A stilbene derivative represented by the following general formula (1). Embedded image (In the formula, X represents a hydrogen atom or a nitrile group, and Y represents an amino acid acyl group.) 2. The stilbene derivative according to claim 1, wherein X in the general formula (1) is a hydrogen atom. 3. The stilbene derivative according to claim 1, wherein X in the general formula (1) is a nitrile group. 4. Y in the general formula (1) is L-α-
The stilbene derivative according to claim 1 or 2, which is an amino acid acyl group. 5. The stilbene derivative according to claim 4, wherein Y in the general formula (1) is an amino acid acyl group derived from threonine or serine. 6. An anti-cancer agent comprising the cis-stilbene derivative according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof.