JPH06306035A - Aphidicolane derivative - Google Patents

Abstract

PURPOSE:To provide a novel 16beta-[2-(acylamino)ethoxy]-3alpha,17,18- trihydroxyaphidicolane having antitumor action. CONSTITUTION:This is a compound expressed by formula I (R is aryl, pyridyl or aryl-substitutedalkenyl), e.g. 16beta-[2-(benzoylamino)ethoxy]-3alpha,17,18- trihydroxyaphidicolane. The compound of formula I can be produced by reacting 3alpha,18-dihydroxy-16beta,17-epoxyaphidicolane with a 2-halogenoethanol, reacting the resultant compound of formula II (X is halogen) with 2,2-dimethoxypropane to obtain a novel 16beta-(2-halogenoethoxy)-17-hydroxy-3alpha,18- isopropylidenoxyaphidicolane of formula III (R<1> is H; R<2> is halogen), protecting the 17-hydroxyl group, introducing 2-(acylamino)ethoxy group to 16beta-site via 3 steps and eliminating the protecting group.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an affidicorane derivative. More specifically, it relates to a novel affidicorane derivative having an antitumor effect.

[0002]

2. Description of the Related Art Aphidicolin, which is a typical Aphidicolin derivative, is a type of fungus, Verticillium.
It was discovered as an antiviral agent obtained from lecanii, is a poorly water-soluble substance having the structure of tetracyclic diterpene tetraol, and shows a high specific inhibitory action on eukaryotic cell-derived DNA polymerase α and δ types. It is used to discriminate the type of DNA, is ineffective for DNA polymerases derived from prokaryotic cells except for special viruses, and is mainly used for analysis of DNA replication and repair mechanism. Further, it is also known to have an antitumor action, an antiviral action, and a plant seed growth inhibitory action, and its use for such purposes has been attempted.

In recent years, attention has been paid to the antitumor activity of aphidicolin, and various derivatives have been synthesized (Japanese Patent Laid-Open Nos. 59-88438, 61-83138, and 62-155232). 62-155239, Nucleic Acid Research No. 1
Volume 7, pp. 6339-6348, 1989, J. Chem. Soc., Perkin T
rans. I 41-46, 1992). In particular, for aphidicolin-17-glycinate, which is a water-soluble ester derivative synthesized as a prodrug, clinical trials are in progress, producing aphidicolin in blood,
It is expected to show antitumor activity, and recently, an optimal clinical test method has been proposed (J. of Pharm. Sc.
i. Vol. 78, No. 5, pp. 399-401, 1989 and J. Am. of the
National Cancer Institute Vol. 83, No. 16, 1991 8
21st of the month).

By the way, there are reports that discuss the structural activity of aphidicolin on the inhibitory action of DNA polymerase α, and various necessity and configurational significance of the four hydroxyl groups contained in aphidicolin have been discussed. , 16β-hydroxyl group is presumed to have a small contribution to the expression of DNA polymerase α inhibitory action (Chem. Pharm.
Bull. (Vol. 35, No. 4, 1641 to 1644, 1987).

However, almost no affidicolane derivative in which only the 16β-hydroxyl group is substituted with another functional group has been reported, and 16β- was introduced for the purpose of protecting the hydroxyl group.
Only methoxy methoxy derivatives are known, 16
The role of the β-position substituent is not clear. Thus, it can be said that the field of affidichorane derivatives having a functional group other than the hydroxyl group at the 16β-position and their biological activities are undeveloped fields.

[0006]

The object of the present invention is to provide 16β
An object is to provide an affidicorane derivative having an amide bond at the -position.

[0007]

According to the present invention, 16β- [2- (acylamino) ethoxy] -3α represented by the following general formula:
Provided is 17,18-trihydroxyaphidicolane.

[Chemical 2] (Wherein R is an aryl group, a pyridyl group or an aryl-substituted alkenyl group)

Such a novel compound is produced by using 3α, 18-dihydroxy-16β, 17-epoxyaffidichorane as a starting material through the following series of steps.

(1) 3α, 18-dihydroxy-16β, 17-epoxyaffidicolane is reacted with 2-halogenoethanol to give 16β- (2-halogenoethoxy) -3α, 17,18-trihydroxyaffidicolane.

[Chemical 3] (X: halogen atom) The reaction between the two, using a large excess of 2-chloroethanol, 2-bromoethanol and the like also serving as a solvent, in the presence of a catalytic amount of an acidic substance, about 0 ~ 100 ℃, Preferably about 10-30 ° C
At the reaction temperature of. At this time, tetrahydrofuran,
It is also possible to dilute and react with a solvent that does not react with the former epoxy group under reaction conditions such as dioxane. Examples of the acidic substance used as a catalyst include p-toluenesulfonic acid, organic acids such as trifluoroacetic acid, hydrochloric acid,
Examples thereof include inorganic acids such as sulfuric acid and π acids such as tetracyanoethylene.

(2) 16β- (2-halogenoethoxy) -3α, 17,18
16- (2-halogenoethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyafidicoranes-reacted with 2,2-dimethoxypropane

[Chemical 4] (Wherein R ¹ = H, R ² = halogen) The reaction between the two is carried out by suspending or dissolving the raw material aphidicolan compound in a solvent that does not interfere with the reaction such as acetone, and On the other hand, about 2 to 100 times the molar amount of 2,2-dimethoxypropane and a catalytic amount of the acidic substance catalyst as used in the above (1) are added, and the amount is about -5 to 40 ° C, preferably about 0 to 20. It is carried out by reacting at ° C.

(3) 16β- (2-halogenoethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyaphidicorane is sequentially reacted with 3,4-dihydro-2H-pyran and potassium phthalimide, 3α, 18-isopropylidenedioxy-
16β- [2- (phthalimido) ethoxy] -17- (tetrahydropyran-2-yloxy) affidicorane

[Chemical 5] (Wherein R ¹ = tetrahydropyran-2-yl group, R ² = phthalimido group) is reacted with hydrazine monohydrate to give 16β- (2-aminoethoxy) -3α, 18- Isopropylidenedioxy-17- (tetrahydropyran-2-yloxy)
Affilicolan

[Chemical 6] (Where R ¹ = tetrahydropyran-2-yl group, R ² = NH ₂ ).
Get

The reaction with 3,4-dihydro-2H-pyran is carried out by dissolving the raw material aphidicolan compound in a solvent inert to the reaction such as dichloromethane or chloroform, and adding about 2 to 10 to the raw material compound. A double molar amount of 3,4-dihydro-2H-pyran and a catalytic amount of an acidic substance catalyst as used in (1) above are added, and the reaction is carried out at about -5 to 40 ° C, preferably about 0 to 20 ° C. It is carried out by The reaction of this reaction product with potassium phthalimide is carried out in an aprotic polar solvent such as N, N-dimethylformamide or dimethylsulfoxide with about 1 to 10 times the molar amount of potassium phthalimide.
It is carried out by reacting at ˜140 ° C., preferably at 70˜100 ° C. Subsequent reaction with hydrazine monohydrate dissolves or suspends the obtained affidicorane compound in an organic solvent such as methanol or ethanol, and the hydrazine monohydrate of about 1 to 10 times is added thereto. It is carried out by adding a hydrate and reacting at about 20 to 100 ° C, preferably about 30 to 50 ° C.

(4) 16β- (2-aminoethoxy) -3α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) afidicorane, with the general formula RCOY (R: aryl group, pyridyl group Or an aryl-substituted alkenyl group, Y: O
16β- [2- (acylamino) ethoxy] -3 by reacting a carboxylic acid (halide) represented by H group or halogen group)
α, 18-Isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) affidicorane

[Chemical 7] (Where R ¹ = tetrahydropyran-2-yl group, R ² = NHCO
R) is obtained, and the protecting group is removed to give the desired product.

[Chemical 8] Get

As the carboxylic acid (halide), benzoic acid, 2-mercaptobenzoic acid, 2-methylbenzoic acid, 3-methoxybenzoic acid, 3-chlorobenzoic acid, 3-cyanobenzoic acid,
4-nitrobenzoic acid, 4-fluorobenzoic acid, nicotinic acid,
Isonicotinic acid, picolinic acid, trans-cinnamic acid or their acid halides are used. Therefore, the R group at the 16β-position which is substituted by reacting these is an aryl group, a pyridyl group or an aryl-substituted alkenyl group which may be unsubstituted or may have a substituent.

When a carboxylic acid is used, N, N'-dicyclohexylcarbodiimide is used as a dehydrating agent for the dehydration condensation reaction with the amino group in an amount of about 1 to 5 times the molar amount of the affidicorane compound. In a solvent such as dimethylformamide, about 1
The condensation reaction with ˜5 times the molar amount of carboxylic acid is carried out at a reaction temperature of about 10-40 ° C. When a carboxylic acid halide is used, the dehydrohalogenation condensation reaction with the amino group is carried out under basic conditions such that pyridine is also used as a solvent to give an affidicorane compound. On the other hand, the condensation reaction with a carboxylic acid halide in a molar amount of about 2 to 10 times is carried out at a reaction temperature of about 0 to 80 ° C, preferably about 20 to 50 ° C.

Then, by stirring in an organic solvent such as methanol or ethanol or a water-containing organic solvent thereof in the presence of the acidic substance catalyst as used in the above (1), at about 0 to 40 ° C., The elimination of the protecting groups 3α, 18-isopropylidene and 17- (tetrahydropyran-2-yl) is carried out.

Although the above-mentioned four kinds of intermediates having a 17- (tetrahydropyran-2-yl) group are a mixture of two kinds of diastereomers, they show one point in thin layer chromatography, None of them was separated in each step and used as the raw material affidicorane compound in the next step.

[0018]

INDUSTRIAL APPLICABILITY The present invention provides a novel aphidicolin derivative having an amide bond at the 16β-position. Also provided is 16β- (2-halogenoethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyafidicorane, which is useful as a synthetic intermediate thereof.

The 16β- [2- (acylamino) ethoxy] afidicorane derivative according to the present invention has an antitumor activity, and its activity generally depends on the chemical structure, but the concentration is generally 0.78 to 3.13 μg. Within the range of / ml, it completely suppresses the growth of mouse tumor cells YAC-1 in vitro.

[0020]

EXAMPLES The present invention will now be described with reference to examples.

Reference Example 3 α, 18-dihydroxy-16β, 17-epoxy affidicolane 650 mg was dissolved in 2-chloroethanol 5 ml, p-toluenesulfonic acid monohydrate 3 mg was added, and at room temperature. Stir for 1 hour. The reaction mixture was poured into 100 ml of ice water, the precipitate was collected by filtration and dried under reduced pressure to obtain 730 mg of powder. This powder was flash chromatographed (particle size 1
Purified with 5-40 μm Merck silica gel 60 40 g, eluent: methanol / chloroform = 1/49) and recrystallized with a diethyl ether-cyclohexane mixed solvent to give 16β- (2-chloroethoxy) -3α, 17, 196 mg of 18-trihydroxyaffidichorane was obtained as colorless needles. Melting point: 136-140 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270 MHz) δ: 0.80 (3H, s), 1.
03 (3H, s), 2.49 (1H, dd J = 3.6,13.2Hz), 2.59 (1H, t J = 6.
9Hz), 2.85 (1H, dd J = 3.5,12.2Hz), 3.61 ~ 3.85 (6H, m),
3.91 (2H, t J = 5.6Hz), 3.93 (1H, b) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 80.0, 76.4, 7
1.9, 64.1, 62.5, 49.3, 45.1, 40.98, 40.55, 40.1, 3
9.8, 34.1, 32.9, 31.7, 27.4, 27.24, 27.15, 25.31,
25.26, 23.5, 18.1, 15.4

Example 1 16β- (2-chloroethoxy) -3α, 17,18-obtained in Reference Example
3.71 g of trihydroxyaffidicholane, 18 m of acetone
It was suspended in ice and cooled on ice. To this suspension, 4.5 ml of 2,2-dimethoxypropane and a catalytic amount of p-toluenesulfonic acid monohydrate were added, and the mixture was stirred for 30 minutes under ice cooling. The reaction mixture was poured into 200 ml of a 2% sodium hydrogen carbonate aqueous solution and then extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was subjected to column chromatography (Wako Gel C-300 500
g, eluent: chloroform) to obtain 16β- (2-chloroethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyafidicorane (2.37 g) as colorless powder. ¹ H NMR (CDCl ₃ , TMS) (270 MHz) δ: 0.73 (3H, s), 0.99 (3H,
s), 1.41 (6H, s), 1.62 (2H, s), 3.21 to 3.67 (9H, m) ¹³ C NMR (CDCl ₃ , TMS) (68MHz) δ: 15.5, 17.4, 19.1, 2
2.6, 24.3, 24.6, 25.0, 26.7, 26.9, 30.0, 31.2, 32.
7, 33.4, 35.2, 39.4, 39.6, 40.4, 44.4, 48.8, 61.
3, 62.5, 68.8, 73.5, 79.9, 98.0

Example 2 2.37 g of 16β- (2-chloroethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyaphidicolane obtained in Example 1 was dissolved in 12 ml of chloroform. To this solution, 1.5 ml of 3,4-dihydro-2H-pyran and a catalytic amount of p-
Toluenesulfonic acid monohydrate was added, and the mixture was stirred at room temperature for 10 minutes. The reaction mixture was poured into ice-cooled 2% aqueous sodium hydrogencarbonate solution (150 ml) and then extracted with chloroform. The chloroform layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was subjected to column chromatography (Merck silica gel 60 400 with a particle size of 40 to 63 μm).
g, eluent: chloroform), and collect fractions exhibiting an Rf value of 0.82 by thin layer chromatography (silica gel 60 Art.5715, manufactured by Merck & Co., Inc .; developing solvent: benzene / ethyl acetate = 2/1) to obtain 16β- 1.49 g of (2-chloroethoxy) -3α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) afidicorane was obtained as a colorless candy-like substance.

After dissolving 1.49 g of this colorless candy-like substance in 15 ml of N, N-dimethylformamide, 3.69 g of potassium phthalimide was added, and the mixture was stirred at 90 to 100 ° C. for 10 hours. The reaction mixture was poured into 150 ml of ice water and then extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. Chloroform (30 ml) was added to the residue, the insoluble matter was removed by filtration, and the filtrate was concentrated to dryness to give 3α, 18-isopropylidenedioxy-16β- [2- (phthalimido) ethoxy] -17- (tetrahydropyran-2. 3.24 g of -yloxy) affidichorane was obtained as a colorless candy-like residue. This material was analyzed by thin layer chromatography (Merck silica gel 6
0 Art.5715, developing solvent: benzene / ethyl acetate = 2/1) R
An f value of 0.67 is shown.

3α, 18-isopropylidenedioxy-16β- [2- (phthalimido) ethoxy] -1 thus obtained
After dissolving 3.24 g of 7- (tetrahydropyran-2-yloxy) affidicolane in 35 ml of ethanol, 3.7 ml of hydrazine monohydrate was added, and the mixture was stirred at room temperature for 2 hours. After filtering the reaction mixture, the filtrate was concentrated to dryness. In the residue,
After 50 ml of water and 50 ml of chloroform were added for liquid separation, the chloroform layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was treated with ether to give 16β- (2-aminoethoxy) -3α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) affidichorane 1.33.
g was obtained as a colorless powder. This substance shows an Rf value of 0.14 by thin layer chromatography (Silica gel 60 Art.5715, manufactured by Merck, developing solvent: methanol / chloroform = 1/9).

Example 3 436 mg of nicotinic acid was dissolved in 7 ml of N, N-dimethylformamide and then N, N'-dicyclohexylcarbodiimide 73
0 mg was added, and the mixture was stirred at room temperature for 3 hours. The precipitate is removed,
16β- (2-aminoethoxy) -3 obtained in Example 2 in the solution
300 mg of α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) afidicolane was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was poured into 100 ml of ice water and extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and then concentrated to dryness. To the residue were added 5 ml of methanol and a catalytic amount of p-toluenesulfonic acid monohydrate, and the mixture was stirred at room temperature for 13 hours. The reaction mixture was neutralized with sodium hydrogen carbonate and then concentrated to dryness. After water and ethyl acetate were added to the obtained residue to separate the layers, the ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. Column chromatography of the residue
(Purified with Merck silica gel 60 10 g having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 7/100),
Further HPLC (Inertosyl PREP-ODS, eluent: methanol /
Purified with water = 7/3), 3α, 17,18-trihydroxy-16β-
[2- (nicotinoylamino) ethoxy] affidichorane 3
2.4 mg was obtained as a colorless powder. ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.77 (3H, s), 0.
99 (3H, s), 3.63 to 3.93 (11H, m), 7.27 (1H, dd J = 5.0Hz),
8.50 (1H, d J = 7.6Hz), 8.63 (1H, d J = 4.9Hz), 9.66 (1H, b) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 15.4, 18.1, 2
3.5, 25.3, 26.0, 27.2, 27.4, 33.1, 34.0, 34.3, 39.
9, 40.0, 40.5, 41.0, 41.7, 49.2, 59.9, 62.5, 71.
9, 76.3, 79.5, 131.3, 135.4, 152.1, 166.0

Example 4 16β- (2-aminoethoxy) -3α, 18-obtained in Example 2
Isopropylidene dioxy-17- (tetrahydropyran-2-
(Yloxy) affidicorane 200 mg was dissolved in pyridine 2 ml, benzoyl chloride 326 μl was added, and the mixture was allowed to stand at room temperature.
Stir for 10 minutes. The reaction mixture was poured into ice water (20 ml) and then extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was subjected to column chromatography (15 g of silica gel 60 manufactured by Merck Co. having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 1/100), and thin layer chromatography (silica gel 60 Art.5715 manufactured by Merck Co., development). Solvent: Methanol /
After collecting the fractions showing Rf value 0.87 with chloroform = 1/9),
It was concentrated to dryness. To the obtained residue, 4 ml of methanol and a catalytic amount of p-toluenesulfonic acid monohydrate were added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was neutralized with sodium hydrogen carbonate and then concentrated to dryness. After water and ethyl acetate were added to the residue for liquid separation, the ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was subjected to column chromatography (10 g of silica gel 60 manufactured by Merck & Co., Inc. having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 1/2).
After purification in 5), the product was recrystallized from methanol to obtain 36 mg of 16β- [2- (benzoylamino) ethoxy] -3α, 17,18-trihydroxyafidicorane as a colorless powder. ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.79 (3H, s), 0.
99 (3H, s), 3.62 to 3.93 (10H, m), 7.43 to 7.46 (3H, m), 8.3
1 ~ 8.34 (2H, m) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 15.5, 18.2, 2
3.6, 25.6, 25.9, 27.29, 27.34, 27.5, 31.7, 33.3, 3
4.2, 40.0, 40.2, 40.7, 41.1, 41.7, 49.3, 60.2, 62.
9, 72.0, 76.4, 79.5, 128.1, 128.7, 131.3, 135.9, 1
67.7

Example 5 432 mg of thiosalicylic acid was added to 4 ml of N, N-dimethylformamide.
After dissolving in the solution, 572 mg of N, N'-dicyclohexylcarbodiimide was added, and the mixture was stirred at room temperature for 2 hours. The precipitate was removed by filtration, and 200 mg of 16β- (2-aminoethoxy) -3α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) affidicorane obtained in Example 2 was added to the filtrate. The mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into 40 ml of ice water and extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and then concentrated to dryness.
The residue was subjected to column chromatography (15 g of silica gel 60 manufactured by Merck Co. having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 1/100), and thin layer chromatography (silica gel 60 Art.5715 manufactured by Merck Co., development). Fractions showing an Rf value of 0.81 were collected with a solvent: methanol / chloroform = 1/9) and then concentrated to dryness. To the obtained residue, 4 ml of methanol and a catalytic amount of p-toluenesulfonic acid monohydrate were added,
The mixture was stirred at room temperature for 5 hours. The reaction mixture was neutralized with sodium hydrogen carbonate and then concentrated to dryness. After water and ethyl acetate were added to the residue for liquid separation, the ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue,
After purification by column chromatography (10 g of silica gel 60 manufactured by Merck Co. having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 1/20), preparative thin layer chromatography (Silica gel 60 Art.13895 manufactured by Merck, Purification with developing solvent: methanol / chloroform = 1/9), 3
30 mg of α, 17,18-trihydroxy-16β- [2- (2-mercaptobenzoylamino) ethoxy] affidicolane was obtained as a colorless powder. ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.79 (3H, s), 1.
01 (3H, s), 3.63 to 3.94 (10H, m), 7.09 to 7.17 (2H, m), 8.02
~ 8.05 (2H, m) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 15.4, 18.1, 1
9.2, 23.5, 25.5, 25.9, 27.2, 27.5, 31.6, 33.2, 34.
1, 39.9, 40.1, 40.6, 41.0, 41.6, 49.2, 60.1, 62.
8, 71.9, 76.3, 79.5, 125.9, 126.7, 128.4, 131.2, 1
34.9, 138.8, 168.3

Example 6 411 mg of trans-cinnamic acid was added to N, N-dimethylformamide.
After dissolving in 4 ml, 572 mg of N, N'-dicyclohexylcarbodiimide was added, and the mixture was stirred at room temperature for 2 hours. The precipitate was removed by filtration, and 200 mg of 16β- (2-aminoethoxy) -3α, 18-isopropylidenedioxy-17- (tetrahydropyran-2-yloxy) afidicolane obtained in Example 2 was added to the filtrate. In addition, the mixture was stirred at room temperature for 2 hours. 40 ml of ice water in the reaction mixture
After pouring in, it was extracted with ethyl acetate. The ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was subjected to column chromatography (particle size 40-6
20 μg of 3 μm Merck silica gel 60, eluent: methanol / chloroform = 1/100), and thin layer chromatography (Merck silica gel 60 Art.5715, developing solvent: methanol / chloroform = 1/9). Fractions showing an Rf value of 0.73 were collected and concentrated to dryness. Methanol was added to the obtained residue.
4 ml and a catalytic amount of p-toluenesulfonic acid monohydrate were added, and the mixture was stirred at room temperature for 15 hours. The reaction mixture was neutralized with sodium hydrogen carbonate and then concentrated to dryness. After water and ethyl acetate were added to the residue for liquid separation, the ethyl acetate layer was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by column chromatography (10 g of silica gel 60 manufactured by Merck & Co., Inc. having a particle size of 40 to 63 μm, eluent: methanol / chloroform = 1/25) and then further purified by HPLC (Inert Syl PR
EP-ODS, eluent: Methanol / water = 4/1), 16β-
75 mg of [2- (cinnamoylamino) ethoxy] -3α, 17,18-trihydroxyafidicolane was obtained as a colorless powder. ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.78 (3H, s), 0.
99 (3H, s), 3.62 to 3.92 (11H, m), 6.94 (1H, d J = 15.8Hz),
7.26 ~ 7.29 (3H, m), 7.47 ~ 7.51 (2H, m), 8.03 (1H, d J = 15.5
Hz) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 15.3, 18.0, 2
3.4, 25.4, 25.8, 27.2, 27.3, 31.6, 33.1, 34.0, 39.
7, 40.0, 40.5, 40.9, 41.1, 49.2, 60.2, 63.0, 71.
8, 76.2, 79.6, 123.0, 127.9, 129.1, 129.5, 135.8,
139.6, 166.2

[Tumor cell proliferation inhibition test] Mouse tumor cell YA
C-1 10 ⁶ cells / ml containing RPMI 1640 medium (10% FCS, 100 U /
ml penicillin and 100 μg / ml streptomycin added) 100 μl was dispensed into each well of a 96-well microplate, and 16β-acylaminoethoxy-3α, 17,18-tri obtained in each of the above-mentioned Examples at various concentrations. Hydroxyaffidichorane (0.5% dimethyl sulfoxide solution was added to the RPMI
100 μl diluted in 1640 medium and adjusted to various concentrations) was added to each well and cultured at 37 ° C. in a 5% CO ₂ incubator for 72 hours. The change in the color of the medium was compared with that of the control to determine the presence or absence of tumor cell proliferation. As a result, the following results were obtained as the concentration at which tumor cell proliferation was completely suppressed. 16β-acylaminoethoxy derivative concentration (μg / ml) Example 3 3.13 〃 4 0.78 〃 5 1.56 〃 6 1.56

Claims (3)

[Claims] 1. A general formula: (Wherein R is an aryl group, a pyridyl group, or an aryl-substituted alkenyl group), and 16β- [2- (acylamino) ethoxy] -3α, 17,18-trihydroxyaffidicolane. 2. 16β- (2-halogenoethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyafidicolin. 3. 16β- (2-halogenoethoxy) -3α, 17,18-
A process for producing 16β- (2-halogenoethoxy) -17-hydroxy-3α, 18-isopropylidenedioxyafidicoranes, which comprises reacting trihydroxyaphidicoranes with 2,2-dimethoxypropane.