JP2000034289A - New compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound showing an inhibitory effect on the genesis of microtubule and useful for an antineoplastic agent, an antifungal agent, or the like. SOLUTION: This compound is expressed by formula I or II [R1 and R2 are each hydroxyl or a group capable of being converted into hydroxyl; R3 is O, S or =NR6 (R6 is an alkyl); R4 and R5 are each an alkyl or an aralkyl] and is e.g. a compound of formula III. The compound of formula I is obtained e.g. by culturing molds belonging to Paecilamyces genus in a medium where a nutrient broth containing 50 wt.% PD agar is prepared using mixed water of purified water with natural seawater in a ratio of 1:1.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel compound having an inhibitory effect on microtubule formation.

[0002]

BACKGROUND OF THE INVENTION Microtubules are small hollow cylindrical structures,
Electron microscopy has confirmed that it is present in all eukaryotic cells. Microtubules are thought to perform two types of interrelated activities. That is, (1) it provides structural support to the cells and acts as a kind of skeleton, and (2) it becomes part of the equipment necessary for some kind of movement.

[0003] The wall of a microtubule is usually composed of thirteen subunits, which are parallel to the long axis of the tube, forming a protofilament. The subunit consists of two proteins, α
It consists of a heterodimer consisting of tubulin and β-tubulin; That is, the microtubule has a tubular structure composed of 13 protofilaments.

[0004] The formation of microtubules in cells is governed by the hydrolysis reaction of guanine nucleotides bound to tubulin at the ends of microtubules, and this dynamic instability (microtubule dynamics) is caused by mitosis and flagella.・ It is known to be important in performing a wide variety of central functions such as hair movement, organelle transport, secretory function, cytoskeleton formation, and biological membrane function [Annual Review of Cell Bio (Annual Reveiw of Cell Biology),
7, 93 (1991); Annual Review of
Biophysics and Biomolecular Structure (Annual Review of Biophysics and Biom)
olecular Structure), 21, 145 (1992)].

[0005] In particular, in the progress of mitosis (M phase) in eukaryotes, changes in microtubule dynamics play a fundamental role in biological phenomena. That is, long and stable interphase microtubules become unstable and disappear in M phase, and short and unstable spindle microtubules appear instead.
This is because eukaryotic cells cannot undergo mitosis unless spindle formation due to the reorganization of microtubules occurs [Science, 246, 622 (1989);
ell), 71, 547 (1992)].

[0006] Drugs that act on microtubules and inhibit mitosis have, with a few exceptions, the ability to bind tubulin, but there are many clinically useful anticancer drugs. Further, it is applied to pharmaceuticals or agricultural chemicals as antifungal agents based on the same mechanism of action. These drugs can be classified into two types, polymerization inhibitors and polymerization accelerators, based on the difference in the mechanism of action on microtubules. Many inhibitors of microtubule polymerization have been found in plants, microorganisms, and marine animals, and improved semisynthetic derivatives thereof have been developed. Clinically applied vinblastines (Cancer Medicine, 1, 78)
2 (1993)].

[0007]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a novel compound having a microtubule formation-inhibiting effect that provides an anticancer effect and an antifungal effect.

[0008]

The object of the present invention is to provide (1) a compound represented by the following general formula [I] or a salt thereof.

[0009]

Embedded image [Wherein, R ₁ and R ₂ represent a hydroxyl group or a group that can be converted to a hydroxyl group, R ₃ represents an oxygen atom, a sulfur atom, ＝ NR ₆ ,
R ₄ and R ₅ represent an alkyl group, an aryl group, or a substituted alkyl group. R ₆ represents an alkyl group. (2) A compound represented by the following general formula [II] or a salt thereof.

[0010]

Embedded image [In the formula, R ₁ and R ₂ represent a hydroxyl group or a group that can be converted to a hydroxyl group. (3) A compound represented by the following formula or a salt thereof. (3) A compound represented by the following formula [III] or a salt thereof.

[0011]

Embedded image Achieved by

Hereinafter, the present invention will be described in detail.

In the general formulas [I] and [II], the group which can be converted into a hydroxyl group represented by R ₁ and R ₂ is a group which can be converted into a hydroxyl group in a living body, for example, a group having 1 to 6 carbon atoms.
(E.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, hexyl) An aryloxy group having 6 to 10 carbon atoms (eg, phenoxy group, naphthoxy group, etc.), an aralkyloxy group having 7 to 12 carbon atoms (eg, benzyloxy group, phenylethyloxy, naphthylmethyloxy group, etc.) ), An alkylcarbonyloxy group having 1 to 6 carbon atoms (eg, acetyloxy group, propionyloxy group, etc.), an arylcarbonyloxy group having 6 to 10 carbon atoms (eg, benzoyloxy group, naphthylcarbonyloxy group, etc.), carbon Aralkylcarbonyloxy of the formulas 7 to 12 Xy group (for example, benzylcarbonyloxy group, phenylethylcarbonyloxy group, etc.). The group convertible to the hydroxyl group represented by R ₁ and R ₂ may have a substituent, and examples of the substituent include a halogen, an alkoxy group, a carboxyl group, and a sulfo group. As R ₁ and R ₂ , a hydroxyl group is preferable.

In the general formula [I], examples of the alkyl group represented by R ₄ and R ₅ include a heptyl group and an octyl group. The alkyl group represented by R ₄ is preferably a heptyl group, and the alkyl group represented by R ₅ is preferably an octyl group. Examples of the aryl group include a phenyl group and a naphthyl group. Further, examples of the substituent in the substituted alkyl group include a halogen, an alkoxy group, a carboxyl group, and a sulfo group.

The alkyl group represented by R ₆ is preferably a lower alkyl group, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl. Groups.

Formulas (I), (II) and (II
The salt of the compound represented by I) is preferably a pharmaceutically acceptable salt, for example, a salt with a base such as an inorganic base, an organic base, or a basic amino acid, an inorganic acid, an organic acid, or an acid such as an acidic amino acid. And salts thereof.

The salt with an inorganic base includes, for example, alkali metal salts (eg, sodium salt, potassium salt, etc.), alkaline earth metal salts (eg, calcium salt, magnesium salt, etc.), aluminum salt, ammonium salt, etc. Examples of the salt with an organic base include salts with trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N'-dibenzylethylenediamine, and the like. Examples of the salt with an acid include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like, and examples of the salt with an organic acid include:
Formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid,
Tartaric acid, maleic acid, citric acid, succinic acid, malic acid,
Salts with methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like, and salts with basic amino acids include, for example, salts with arginine, lysine, ornithine, and the like. And the like, for example, salts with aspartic acid and glutamic acid. As such a salt, a sodium salt, a potassium salt, an ammonium salt, a hydrochloride, an acetate, an oxalate, a citrate and the like are preferable, and a sodium salt, a potassium salt, a hydrochloride and the like are more preferable.

The compound represented by the formula [III] of the present invention is
A filamentous fungus of the genus aecilomyces is cultured, and isolated and
It can be obtained by purification. The compounds represented by the general formulas [I] and [II] can be synthesized by a usual alkylation method, salt exchange method and the like.

[0019]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples.

Example 1 [Culture of a producing bacterium and extraction and isolation of a compound] 1/2 nutrition PD
The medium is adjusted with a mixture of purified water and natural seawater (1: 1),
100 mL was put into a 500 mL Erlenmeyer flask (autoclave sterilization), and a filamentous fungus of the Paecilomyces family was inoculated from the strand. In 15 flasks (1.5 L total), 2
The culture was allowed to stand at 0 ° C. for 3 weeks, and filtered to obtain 11.1 g (wet weight) of cells. The cells were extracted three times with methanol, and the extract (1.5 g) obtained by concentrating to dryness was suspended in water and adsorbed on an ODS column (50 g).
After washing with a 25%, 50%, and 75% methanol-water mixture of each mL, the mixture was extracted with 100% methanol (300 mL) and concentrated (350.6 mg was obtained after concentration). This was concentrated using a benzene-acetone (11: 1) mixed solvent and a silica gel column to concentrate the biologically active substance (206.3 m).
L), and then 8.5 mg of a compound (compound represented by the formula [III]) was isolated using a benzene-acetone (98: 2) mixed solvent and a silica gel column.

The physical properties of the compound represented by the formula [III] are shown in Table 1 below.

[0022]

[Table 1] Table 2 shows the spectrum data for determining the structure of the compound represented by the formula [III]. Formula 3 shows the positions in Table 2.

[0023]

[Table 2]

[0024]

Embedded image FIG. 1 shows the microtubule polymerization inhibitory activity of the obtained compound represented by the formula [III].

The microtubule protein used was obtained by performing two cycles of polymerization-standing polymerization from pig brain and purifying it.
The microtubule polymerization inhibitory activity was determined by the turbidity method at 37 ° C. and 400 n
The change in absorption at m was observed. The protein concentration is 1.3m
g / mL. In order to confirm the presence or absence of depolymerization inhibitory activity, 4 mM of CaCl ₂ was added 20 minutes after the start of polymerization.
No depolymerization inhibitory activity was observed.

[0026]

The compounds of the present invention exhibit a microtubule formation inhibitory effect and are useful as antitumor and antifungal agents.

[Brief description of the drawings]

FIG. 1 is a drawing explaining the microtubule polymerization inhibitory activity of a compound represented by the formula [III] obtained in Example 1.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Soga Takahashi 1 Sakuracho, Hino-shi, Tokyo Konica Corporation (72) Inventor Kenichiro Okaba 1 Sakuracho, Hino-shi, Tokyo Konica Corporation F-term (reference) 4C071 AA04 AA07 BB01 BB06 CC12 EE05 FF17 GG01 HH05 LL01 4C086 AA02 AA03 GA02 MA01 MA04 NA14 ZB26 ZB35 ZC02

Claims (3)

[Claims] 1. A compound represented by the following general formula [I] or a salt thereof. Embedded image [Wherein, R ₁ and R ₂ represent a hydroxyl group or a group that can be converted to a hydroxyl group, R ₃ represents an oxygen atom, a sulfur atom, ＝ NR ₆ ,
R ₄ and R ₅ represent an alkyl group, an aryl group, or a substituted alkyl group. R ₆ represents an alkyl group. ] 2. A compound represented by the following general formula [II] or a salt thereof. Embedded image [In the formula, R ₁ and R ₂ represent a hydroxyl group or a group that can be converted to a hydroxyl group. ] 3. A compound represented by the following formula [III] or a salt thereof. Embedded image