JPH069475A - Aphidicolan derivative and its production - Google Patents

Abstract

PURPOSE:To provide a novel compound having an antitumor action. CONSTITUTION:The compound of formula I (R is 2-5C alkylene; X is halogen, OH, cyano, lover alkoxy). For example, 3alpha,17,18-trihydroxy-16beta-(3- hydroxypropoxy)aphidicolan. The compound of formula I is obtained by reacting 3,18-dihydroxy-16,17-epoxyaphidicolan with a compound of formula: XROH.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an affidicorane derivative and a method for producing the same. More specifically, it relates to a novel affidicorane derivative having an antitumor effect and a method for producing the same.

[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
(EN) It is intended to provide an affidicorane derivative having a functional group other than a hydroxyl group at the -position and a method for producing the same.

[0007]

According to the present invention, 16β-substituted alkoxy-3α, 17,18-trihydroxyafidicolane represented by the following general formula is provided.

[Chemical 3] (Wherein R is an alkylene group having 2 to 5 carbon atoms, X is a halogen atom, a hydroxyl group, a cyano group or a lower alkoxy group) Among such novel compounds, 3α, 17,18-trihydroxy-16β- Hydroxyalkoxy aphidicolin is
3α, 18-dihydroxy-16β, 17-epoxy affidicolane and lower alkanediols such as ethylene glycol, 1,3-propanediol, 1,4-butanediol, 1,5-
It is produced by a method of reacting pentanediol or the like.

Raw material compound 3α, 18-dihydroxy-16
β, 17-epoxy aphidicolin

[Chemical 4] Is a known compound, and 3α, 16β, 18-trihydroxy-17-p-toluenesulfonyloxyaphidicorane described in JP-A-59-88438 can also be reacted with a basic substance. Obtained (see Japanese Patent Application No. 3-237489). The reaction between this raw material compound and the lower alkanediol is performed at about 0 to 100 ° C, preferably in the presence of a catalytic amount of p-toluenesulfonic acid monohydrate using a large excess of the lower alkanediol which also serves as a solvent. Is performed at about 10-30 ° C.
At this time, the reaction can be performed by diluting with a solvent such as tetrahydrofuran or dioxane that does not react with each raw material under the reaction conditions.

The thus obtained substance containing 3α, 17,18-trihydroxy-16β-hydroxyalkoxy aphidicorane as a main component is directly purified by column chromatography and recrystallization to obtain a pure object. It can be introduced into a pure product or via a derivative.

In the former case, a pure target product can be obtained by a purification method in which silica gel column chromatography is usually carried out, followed by recrystallization with ethyl acetate, methylene chloride or the like.

In the latter case, the substance containing the above main component is added with about 5 to 100 times the molar amount of 2,2-dimethoxypropane, and a catalytic amount of p-toluenesulfonic acid is added in a solvent such as acetone. 17-hydroxy-16β-hydroxyalkoxy-3α, 18-isopropylidenedioxyaffidicolane is reacted in the presence of a monohydrate or the like at about -5 to 40 ° C, preferably about 0 to 20 ° C.

[Chemical 5] After being converted to a substance containing as a main component, this substance is dissolved in pyridine, and an acylating agent such as benzoyl halide, acetyl halide, benzoic anhydride, acetic anhydride and the like in an amount of about 2 to 10 times and about 0. Reaction at a temperature of ~ 30 ℃, 17-acyloxy-16β-acyloxyalkoxy-3α, 18-isopropylidenedioxyafidicorane

[Chemical 6] (Wherein R ′ is a lower alkyl group or a phenyl group).

Then, about 70-9% in about 70-90% acetic acid aqueous solution.
By heating to 0 ° C. to remove the isopropylidene group, and then removing the acyl group with an aqueous alkali hydroxide solution,
A pure object can be obtained.

Similar to the reaction of 3α, 18-dihydroxy-16β, 17-epoxyaffidichorane with a lower alkanediol, which can be said to be a hydroxy-substituted lower alcohol,
A lower alcohol substituted with a halogen atom, a cyano group or a lower alkoxy group can also be reacted with the 16β, 17-epoxy group.

That is, 3α, 18-dihydroxy-16β, 17-epoxyaffidichorane has the general formula XROH (wherein R is an alkylene group having 2 to 5 carbon atoms, X is a halogen atom, a hydroxyl group or a cyano group). Or a lower alkoxy group) by reacting a compound represented by the general formula

[Chemical 7] 16β-Substituted alkoxy-3α, 17,18-trihydroxyaffidicholanes of the formula (where R and X are as defined above) can be obtained.

As a compound in which X is a halogen atom,
Examples include 2-chloroethanol, 2-bromoethanol, 2-fluoroethanol, 3-chloropropanol, 3-fluoropropanol, and the like, and compounds where X is a cyano group include ethylene cyanohydrin and 3-cyanopropanol. Examples of the compound in which X is a lower alkoxy group include 2-methoxyethanol, 2-ethoxyethanol, 3-ethoxypropanol and the like.

As in the case of the lower alkanediol, the reaction between the two is carried out in the presence of a catalytic amount of an acidic substance using a large excess of these substituted alcohols which also serve as a solvent.
It is carried out at a reaction temperature of -100 ° C, preferably about 0-30 ° C.
At this time, the reaction can be performed by diluting it with a solvent such as tetrahydrofuran or dioxane that does not react with the former epoxy group. Examples of the acidic substance used as a catalyst include organic acids such as p-toluenesulfonic acid and trifluoroacetic acid, inorganic acids such as hydrochloric acid and sulfuric acid, and π acids such as tetracyanoethylene.

[0017]

INDUSTRIAL APPLICABILITY The present invention provides a novel affidicorane derivative having a halogenoalkoxy group, a hydroxyalkoxy group, a cyanoalkoxy group or an alkoxyalkoxy group at the 16β-position. These derivatives have antitumor activity, and the degree of their activity depends on the chemical structure, but generally within the concentration range of about 0.2 to 1.6 μg / ml,
Completely suppresses the growth of mouse tumor cells YAC-1 in vitro. In addition, a dose of about 25 to 300 mg / kg generally shows a life-prolonging effect or a curative effect on mice transplanted with Ehrlich's ascites tumor, and thus it can be regarded as a promising cancer drug. Furthermore, the compound having a 16β-halogenoalkoxy group can be easily led to another derivative by utilizing the halogen group, and thus it is also useful as a raw material for synthesizing another affidicorane derivative expected to have an antitumor effect. Is.

[0018]

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

Example 1 3α, 16β, 18-trihydroxy-17-p-toluenesulphonyloxyaphidicolin was added to a solution prepared by dissolving 2.7 g of potassium hydroxide in 80 ml of methanol while stirring under ice cooling.
After adding 8 g and stirring for 5 minutes, the mixture was further stirred at room temperature for 1 hour.
After filtering the reaction mixture, the filtrate is concentrated under reduced pressure at 20 ° C or lower.
Water was added to the residue, and the mixture was neutralized with a 10% aqueous citric acid solution, extracted with ethyl acetate, the extract was washed with water, dried (anhydrous magnesium sulfate), and concentrated to dryness. The residue was treated with isopropyl ether to give 3.67 g of 3α, 18-dihydroxy-16β, 17-epoxy aphidicorane (melting point 138 to 145 ° C) as a colorless powder.

The identity of this compound was determined by other synthetic methods.
J. Chem. Soc., Perkin Trans. I page 1477 (1988) by comparing with the data of the NMR spectrum.

2 mg of p-toluenesulfonic acid monohydrate was added to a solution prepared by dissolving 3.6 g of the obtained 3α, 18-dihydroxy-16β, 17-epoxyafidicolane in 40 ml of 1,3-propanediol. After addition, the mixture was stirred at room temperature for 1 hour, and the reaction mixture was mixed with 50 ml of 1% aqueous sodium hydrogen carbonate solution and 600 ml of saturated saline solution.
It was poured into a mixed aqueous solution of.

The precipitate was collected by filtration, dissolved in 300 ml of ethyl acetate, washed with 200 ml of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was flash chromatographed (Merck silica gel with particle size 15-40 μm).
60 150g, eluent: Methanol / chloroform = 8/92), then recrystallize with methylene chloride to give 3α, 17,18-trihydroxy-16β- (3-hydroxypropoxy) affidicorane crude crystals 744mg Got

Example 2 To a solution of 744 mg of the crude crystal obtained in Example 1 in 3.5 ml of acetone, 1.0 ml of 2,2-dimethoxypropane and 1 mg of p-toluenesulfonic acid monohydrate were added. , Under ice cooling 1
After stirring for 5 minutes, the reaction mixture was poured into 100 ml of an ice-cooled 1% aqueous sodium hydrogen carbonate solution and extracted with 100 ml of ethyl acetate.

The organic layer was washed with water (100 ml × 2 times), dried (anhydrous sodium sulfate) and concentrated under reduced pressure, and the residue was subjected to flash chromatography (30 g of Merck silica gel 60, eluent: methanol / chloroform = 3/97). ), To obtain 489 mg of a colorless candy-like substance containing 17-hydroxy-16β- (3-hydroxypropoxy) -3α, 18-isopropylidenedioxyafidicorane.

To a solution of this candy-like substance dissolved in 4.5 ml of pyridine, 1.3 ml of benzoyl chloride was added, and after stirring at room temperature for 15 minutes, the reaction mixture was poured into 90 ml of ice water, and 90 ml of diethyl ether was added. It was extracted with.

The organic layer was washed with water (75 ml × 2 times), dried (anhydrous sodium sulfate) and concentrated under reduced pressure, and the residue was subjected to flash chromatography (using 40 g of silica gel 60 from Merck, eluent: ethyl acetate / benzene = 3 / After purification in 97), the crystals were recrystallized from methanol to obtain 303 mg of 17-benzoyloxy-16β- (3-benzoyloxypropoxy) -3α, 18-isopropylidenedioxyafidicorane as colorless needle crystals. Melting point: 124-128 ° C ¹ H NMR (CDCl ₃ , TMS) (270 MHz) δ: 0.71 (3H, s), 0.95 (3H,
s), 1.39 (3H, s), 1.40 (3H, s), 3.22 (1H, d J = 12.2Hz), 3.
54 to 3.68 (4H, m), 4.31 (2H, ABq J = 12.2Hz), 4.44 (1H, t J
= 6.27Hz), 4.45 (1H, t J = 6.27Hz), 7.34 to 7.57 (6H, m),
7.96 to 8.05 (4H, m) ¹³ C NMR (CDCl ₃ , TMS) (68MHz) δ: 15.4, 17.4, 19.1, 2
2.5, 24.3, 24.4, 25.6, 26.6, 26.9, 29.6, 29.7, 31.
4, 32.5, 33.3, 35.2, 39.5, 39.7, 40.4, 48.6, 57.
3, 62.2, 65.5, 68.8, 73.4, 77.6, 97.9, 128.3, 128.
4, 129.5, 129.6, 130.2, 130.4, 132.7, 132.9, 166.5

Example 3 200 mg of 17-benzoyloxy-16β- (3-benzoyloxypropoxy) -3α, 18-isopropylidenedioxyaphidicolane obtained in Example 2 was dissolved in 3 ml of 80% aqueous acetic acid solution. After stirring at 80 ° C for 1 hour, the mixture was concentrated to dryness under reduced pressure. To the solution obtained by dissolving the residue in 3 ml of methanol, 0.5 ml of 4N potassium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 2 hours, and then the reaction mixture was poured into 30 ml of saturated saline, and 50 ml
Was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness at room temperature.

Ethyl acetate (10 ml) was added to the obtained residue, and the mixture was sonicated, and the precipitate was collected by filtration to obtain 82 mg of 3α, 17,18-trihydroxy-16β- (3-hydroxypropoxy) affidicolane. Obtained as a colorless powder. Melting point: 176-178 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.80 (3H, s), 1.
02 (3H, s), 3.61 ~ 4.12 (9H, m), 5.58 (1H, t J = 5.9Hz), 5.
75 (1H, dd J = 3.1,7.6Hz), 5.92 (1H, t J = 5.3Hz), 6.19 (1
H, d J = 2.6Hz) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 79.1, 76.4, 7
1.9, 63.3, 59.8, 58.2, 49.2, 41.0, 40.6, 40.1, 40.
0, 34.1, 33.1, 31.7, 27.4, 27.2, 27.1, 25.5, 23.
5, 18.1, 15.4

Example 4 The purified product by flash chromatography of Example 1 was recrystallized with ethyl acetate to obtain 340 mg of 3α, 17,18-trihydroxy-16β- (3-hydroxypropoxy) affidichorane. The melting point, ¹ H NMR spectrum and ¹³ C NMR spectrum of this product were completely in agreement with those obtained in Example 3.

Example 5 1 g of 3α, 18-dihydroxy-16β, 17-epoxyaphidicolane was dissolved in a mixed solution of 9 ml of ethylene glycol and 2 ml of tetrahydrofuran, and 3 mg of p-toluenesulfonic acid monohydrate was added. Was added, and the mixture was stirred at room temperature for 2 hours.
The reaction mixture was ice-cooled with 0.5% aqueous sodium hydrogen carbonate solution.
After pouring into 20 ml, the mixture was extracted with 100 ml of ethyl acetate. The ethyl acetate layer was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wako Gel C-300 5
0 g, eluent: methanol / chloroform = 1/19) and then recrystallize with ethyl acetate to give 3α, 17,18-trihydroxy-16β- (2-hydroxyethoxy) affidicorane 220 m
g was obtained as colorless crystals. Melting point: 160-167 ° C. ¹ H NMR (pyridine-d ₅ , TMS) (270 MHz) δ: 0.80 (3 H, s), 1.
20 (3H, s), 3.67 to 3.98 (9H, m) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 79.4, 76.3, 7
1.9, 63.3, 62.9, 62.3, 49.3, 41.0, 40.6, 40.1, 34.
1, 33.2, 31.7, 27.4, 27.2, 25.9, 25.6, 23.5, 18.
1, 15.4

Example 6 200 mg of 3α, 18-dihydroxy-16β, 17-epoxyaffidicholane was dissolved in a mixed solution of 1 ml of 1,4-butanediol and 0.1 ml of tetrahydrofuran, and p-toluenesulfonic acid was added. 3 mg of monohydrate was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into ice-cooled 0.5% aqueous sodium hydrogencarbonate solution (20 ml) and then extracted with ethyl acetate (100 ml).
The ethyl acetate layer was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wako Gel C
-300 50 g, eluent: methanol / chloroform = 1/19) and then further purification by HPLC (Inertosyl PREP-ODS, eluent: methanol / water = 4/1). Recrystallize with ethyl, 3α, 17,18-trihydroxy-16β-
36 mg of (4-hydroxybutoxy) afidicorane was obtained as colorless crystals. Melting point: 161-164 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270 MHz) δ: 0.80 (3H, s), 1.
03 (3H, s), 3.63 to 4.03 (9H, m) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 79.0, 76.4, 7
1.9, 63.6, 62.3, 60.9, 49.2, 41.0, 40.6, 40.1, 39.
9, 34.1, 33.1, 31.7, 30.9, 28.0, 27.4, 27.3, 27.
1, 25.4, 25.4, 23.5, 18.1, 15.4 (hereinafter margins)

Example 7 3 mg of p-toluenesulfonic acid monohydrate was added to a solution prepared by dissolving 650 mg of 3α, 18-dihydroxy-16β, 17-epoxyaphidicolane in 5 ml of 2-chloroethanol, and then at room temperature. It was stirred 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 8 3 mg of p-toluenesulfonic acid monohydrate was added to a solution prepared by dissolving 640 mg of 3α, 18-dihydroxy-16β, 17-epoxyaphidicolane in 4 ml of 2-bromoethanol, and at room temperature. It was stirred 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 850 mg of powder. This powder was flash chromatographed (particle size 1
Purified with 37-40 g of 5-40 μm Merck silica gel 60, eluent: methanol / chloroform = 1/49), and then recrystallized with a mixed solvent of diethyl ether-cyclohexane to give 16β- (2-bromoethoxy) -3α, 17, 29 mg of 18-trihydroxyaffidichorane was obtained as colorless needles. Melting point: 128-130 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.80 (3H, s), 1.
04 (3H, s), 2.51 (1H, dd J = 3.6,12.9Hz), 2.60 (1H, t J = 6.
9Hz), 2.86 (1H, dd J = 3.3,12.5Hz), 3.60 (2H, t J = 5.9H
z), 3.67 to 3.86 (4H, m), 3.95 (1H, b), 3.97 (2H, t J = 5.9H
z) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 80.1, 76.3, 7
1.9, 64.1, 62.3, 49.2, 41.0, 40.6, 40.1, 39.8, 34.
1, 33.6, 32.9, 31.7, 27.4, 27.22, 27.15, 25.2, 23.
5, 18.1, 15.4

Example 9 3 mg of p-toluenesulfonic acid monohydrate was added to a solution prepared by dissolving 100 mg of 3α, 18-dihydroxy-16β, 17-epoxyafidicolane in 1 ml of 2-fluoroethanol, and the mixture was allowed to stand at room temperature. It was stirred for 2 hours. The reaction mixture was poured into 30 ml of ice water and then extracted with 30 ml of ethyl acetate. 10 ethyl acetate layers
The extract was washed twice with 0 ml of water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (Wako Gel C-300 10 g, eluent: methanol / chloroform = 3/100), and further HPLC (Inertosyl PREP-ODS, eluent: methanol / water = 4/1). ), And then recrystallized from ethyl acetate to give 16β- (2-fluoroethoxy) -3α, 17,18-trihydroxyaffidichorane 6mg.
Was obtained as a colorless powder. Melting point: 147 to 152 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270 MHz) δ: 0.80 (3H, s), 1.
02 (3H, s), 3.63 to 4.01 (7H, m), 4.62 (2H, dt J _HF = 47.8Hz,
J _HH = 4.29Hz) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 84.2 (d J _CF = 16
7.3Hz), 79.7, 76.3, 71.9, 64.2, 61.4 (d J _CF = 19.5H
z), 49.3, 41.0, 40.6, 40.1, 39.6, 34.0, 32.9, 31.
7, 27.4, 27.24, 27.17, 25.6, 25.3, 23.5, 18.1, 15.
Four

Example 10 To a solution of 500 mg of 3α, 18-dihydroxy-16β, 17-epoxyaphidicolane dissolved in a mixed solution of 5 ml of ethylene cyanohydrin and 0.7 ml of tetrahydrofuran, 128 mg of tetracyanoethylene as a catalyst was added. , At room temperature 1
It was stirred for 4 hours. The reaction mixture was poured into 100 ml of ice water and extracted with 100 ml of ethyl acetate. The ethyl acetate layer was washed twice with 100 ml of water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (Wakogel C-300 30 g, eluent: methanol / chloroform = 3/100), and recrystallized from ethyl acetate to give 16β- (2-cyanoethoxy)-. 50 mg of 3α, 17,18-trihydroxyaffidichorane was obtained as colorless crystals. Melting point: 181-184 ° C ¹ H NMR (pyridine-d ₅ , TMS) (270MHz) δ: 0.79 (3H, s), 1.
03 (3H, s), 2.70 (2H, t J = 6.27Hz), 3.62 ~ 3.89 (6H, m),
3.93 (1H, b) ¹³ C NMR (pyridine-d ₅ , TMS) (68MHz) δ: 119.7, 80.3, 7
6.4, 71.9, 64.3, 57.4, 49.3, 41.0, 40.5, 40.1, 39.
6, 34.1, 33.0, 31.7, 27.4, 27.2, 27.1, 25.4, 25.
3, 23.5, 19.7, 18.1, 15.4

Example 11 3 mg of p-toluenesulfonic acid monohydrate was added to a solution of 100 mg of 3α, 18-dihydroxy-16β, 17-epoxyafidicolane in 1 ml of 2-methoxyethanol, and the mixture was allowed to stand at room temperature. It was stirred for 1 hour. The reaction mixture was poured into ice-cooled 0.5% aqueous sodium hydrogen carbonate solution (30 ml), and then ethyl acetate was added.
Extracted with 50 ml. The ethyl acetate layer was washed with 100 ml of water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (Wako gel C-300 10 g, eluent: methanol / chloroform =
1/50) to obtain 20 mg of 3α, 17,18-trihydroxy-16β- (2-methoxyethoxy) afidicolane as a colorless candy-like substance. ¹ H NMR (CDCl ₃ , TMS) (90 MHz) δ: 0.68 (3H, s), 0.94 (3H,
s), 3.35 (3H, s), 3.20 to 3.80 (9H, m)

The possibility that the 16-position of the obtained affidicorane derivative is in the α-form cannot be denied, but it can be confirmed that it is in the β-form from the stereochemistry of the starting compounds and various analytical data. It is believed that there is.

[Tumor cell growth inhibition test] Mouse tumor cell YA
C-1 10⁶RPMI 1640 medium containing cells / ml (10% FCS, 100U /
ml penicillin and 100 μg / ml streptomycin
Add 100 μl to each well of 96-well microplate
And 16β-substitution obtained in each of the above examples at various concentrations.
Alkoxy-3α, 17,18-trihydroxy affidicola
Solution (5% dimethyl sulfoxide solution in RPMI 1640 medium
100 μl diluted in each well and adjusted to various concentrations)
In addition to, 37 ℃, 5% CO₂72 hours in the incubator
Cultured. Compare the change in color of the medium with the control,
As a result of determining the presence or absence of tumor cell proliferation, tumor cell proliferation
As a concentration that completely suppresses
It was16β-Substituted alkoxy derivative Concentration (μg / ml)  Example 3 0.78 〃 5 0.20 〃 6 0.78 〃 7 0.39 〃 8 1.56 〃 9 0.39 〃 10 0.39 〃 11 0.78

[Physiological activity test] About 10 ⁵ Ehrlich ascites tumor cells grown in the abdominal cavity of ICR mice / mouse
Samples were implanted intraperitoneally into week-old male ICR mice (10 mice per group).
100 mg / (of 16 β-hydroxyalkoxy derivative obtained in Examples 3 and 5 was well pulverized and then well mixed with physiological saline containing 1-2 drops of Tween80 to prepare 10 mg / ml)
The dose of kg body weight was continuously administered intraperitoneally once a day for 7 days from the 0th day, and the survival days were observed.

The results obtained are shown in FIGS. 1 and 2 below. In the control group, the same volume of lysate alone was used,
It was administered under the same conditions. From this result, it is clear that the novel affidicorane derivative according to the present invention exhibits a life prolonging effect as compared with the control administration group.

[Brief description of drawings]

FIG. 1 is a graph showing the results of a physiological activity test of the 16β-hydroxypropoxy derivative obtained in Example 3.

FIG. 2 is a graph showing the results of a physiological activity test of the 16β-hydroxyethoxy derivative obtained in Example 5.

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[Procedure amendment]

[Submission date] May 21, 1993

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] Claim 2

[Correction method] Change

[Correction content]

[Chemical 2] And reacting a compound represented by the general formula XROH (wherein R is an alkylene group having 2 to 5 carbon atoms, and X is a halogen atom, a hydroxyl group, a cyano group or a lower alkoxy group). The method for producing 16β-substituted alkoxy-3α, 17,18-trihydroxyafidicolane according to claim 1 .

Claims (6)

[Claims] 1. A general formula: (Wherein R is an alkylene group having 2 to 5 carbon atoms, X is a halogen atom, a hydroxyl group, a cyano group or a lower alkoxy group) 16β-substituted alkoxy-3α, 17,18-
Trihydroxy aphidicolin. 2. 3α, 18-dihydroxy-16β, 17-epoxy affidichorane has the general formula XROH (where R is a carbon number of 2-5).
Is an alkylene group of X, and X is a halogen atom, a hydroxyl group, a cyano group or a lower alkoxy group), and a compound represented by the general formula: (Wherein R and X are the same as defined above), and a method for producing 16β-substituted alkoxy-3α, 17,18-trihydroxyafidicorane. 3. 17-Benzoyloxy-16β-benzoyloxyalkoxy-3α, 18-isopropylidenedioxyaphidicolane. 4. 17-Hydroxy-16β, which comprises reacting 3α, 17,18-trihydroxy-16β-hydroxyalkoxy aphidicolin with 2,2-dimethoxypropane.
-A method for producing hydroxyalkoxy-3α, 18-isopropylidenedioxyafidicorane. 5. A 17-acyloxy-16β-acyloxyalkoxy-3α, 18-characterized in that 17-hydroxy-16β-hydroxyalkoxy-3α, 18-isopropylidenedioxyaphidicolin is reacted with an acylating agent. Process for producing isopropylidenedioxyafidicolin. 6. A 3α, 17,1 characterized in that a 17-acyloxy-16β-acyloxyalkoxy-3α, 18-isopropylidenedioxyaphidicolin is hydrolyzed.
A method for producing 8-trihydroxy-16β-hydroxyalkoxy aphidicorane.