JP2005314236A - New quassinoid-based compound and use thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a new anticancer agent since a conventional quassinoid-based compound is not sufficient as an anticancer agent so far in terms of toxicity and efficacy and a new anticancer agent is required. <P>SOLUTION: The medicine, especially the anticancer agent comprises bruceanol A reduction body of a physiologically active substance, bruceanol B reduction body of a physiologically active substance, bruceantinoside B reduction body of a physiologically active substance, ailanthone reduction body of a physiologically active substance or its pharmaceutically permissible salt as an active ingredient. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a novel quasinoid compound or a pharmacologically acceptable salt thereof, and a medicament, cell growth inhibitor or anticancer agent containing the compound as an active ingredient.

Bruceanthin has been extracted and isolated as an active ingredient of Brucea antidiscenterica Mill., A long-necked plant that has long been used for cancer treatment in Ethiopia (Non-patent Document 1). With this as an opportunity, bruceanthalin, brucein B, dehydrobruseanthin, brucein D, brucein E, bruceoside A, dehydroairantinone, glaucarbinone, ilanton, yadangioside N, yadangioside O, etc. from the same plant or the same genus Organic compounds and quasinoid compounds that are derivatives thereof have been found. For example, Non-Patent Document 2 and Non-Patent Document 3 also report anticancer effects of these compounds. However, these quasinoid compounds have not been sufficient as therapeutic anticancer agents in terms of toxicity, efficacy and the like (Non-patent Document 4).
Non-patent document 5 describes the production of ailanthone reductant, but no pharmacological activity has been found.

S. M.M. Kupchan et al. , J .; Org. Chem. , 38, 178 (1973) S. M.M. Kupchan et al. , J .; Org. Chem. , 40, 648 (1975) S. M.M. Kupchan et al. , J .; Org. Chem. , 40, 654 (1975) J. et al. C. Arseneau et al. , Investigative New Drugs, 1,239 (1983) H. Tada et al. , Eur. J. et al. Med. Chem. , 26, 345 (1991)

  As described above, there are no known quasinoid compounds that are satisfactory in clinical practice, and there is a need for quasinoid compounds that can be used as new therapeutic agents.

  In order to solve the above-mentioned problems, the present inventors have conducted intensive research and found that each reduced form of bruceanol A, bruceanol B, brucetinside B and ailanthone, which are plant components, becomes a novel antitumor agent with reduced toxicity. The headline and the present invention have been completed.

That is, the present invention
(1) The following formula (I)

A bioactive substance bruceanol A reduced form represented by the following formula (II)

A physiologically active substance bruceanol B reduced form represented by the formula:
(2) The following formula (III)

Or a pharmacologically acceptable salt thereof, or a physiologically active substance bruceantinoside B reductant represented by:
(3) A medicament comprising the compound according to (1) or (2) above or a pharmacologically acceptable salt thereof as an active ingredient;
(4) A cell growth inhibitor comprising the compound according to (1) or (2) above or a pharmacologically acceptable salt thereof as an active ingredient;
(5) An anticancer agent comprising the compound according to (1) or (2) above or a pharmacologically acceptable salt thereof as an active ingredient;

(6) The following formula (IV)

A pharmaceutical comprising, as an active ingredient, a reduced form of the physiologically active substance ailanthone represented by the formula:
(7) a cell growth inhibitor comprising as an active ingredient the compound according to (6) or a pharmacologically acceptable salt thereof;
(8) An anticancer agent comprising as an active ingredient the compound according to (6) or a pharmacologically acceptable salt thereof;
About.

  The bruceanol A reductant, bruceanol B reductant, bruceantinoside B reductant or ailanthone reductant of the present invention, or a pharmacologically acceptable salt thereof has a cell growth inhibitory action and has anticancer agents, particularly multidrug resistance. It is useful as a chemotherapeutic agent for the indicated cancer.

  The bruceanol A reductant of the present invention has the structural formula of the above formula (I), the bruceanol B reductant has the structural formula of the above formula (II), and the bruceantinoside B reductant has the structural formula of the above formula (III). The novel quasinoid compounds represented by formula (I), formula (II) or formula (III) can be produced, for example, by subjecting known quasinoid compounds which are plant components to a reduction reaction. In addition, an ailanthone reductant can also be produced, for example, under the following reducing conditions.

  Specifically, bruceanol A, bruceanol B or ailanthone can be dissolved in a solvent such as methanol, and as a catalytic hydrogen reduction catalyst, for example, Pd / C (5%; palladium supported on activated carbon) is added, The reaction was stirred for 24 hours under a hydrogen atmosphere, the catalyst was filtered off, the solvent was distilled off, the residue was purified by silica gel column chromatography using a mixed solvent of ethyl acetate-diethyl ether, and recrystallized as necessary to regenerate bruceanol A. , Bruceanol B reductant or ailanthone reductant is obtained.

  Further, bruceantinoside B is dissolved in a solvent that can dissolve B, such as methanol, and, for example, Pd / C (5%; palladium supported on activated carbon) is added as a catalytic hydrogen reduction catalyst, followed by stirring and reaction in a hydrogen atmosphere for 24 hours. After removing the catalyst by filtration, the solvent was distilled off, and the residue was purified using preparative HPLC (high resolution liquid chromatography) using a mixed solvent of methanol and water, and recrystallized as necessary to obtain a reduced form of bruceantinoside B. It is done.

  The present invention also includes stereoisomers of quasinoid compounds represented by the above formula (I), formula (II), formula (III), and formula (IV), and these stereoisomers are included in the use of the present invention. Can also be used.

  In the present invention, the pharmacologically acceptable salt is not particularly limited as long as it is a salt that can be usually taken. Examples thereof include salts with alkali metals, that is, sodium salts, potassium salts and the like.

  The medicament comprising the bruceanol A reduced form, bruceanol B reduced form, bruceantinoside B reduced form or ailanthone reduced form of the present invention, or a pharmacologically acceptable salt thereof, can preferably be used as a cell growth inhibitor. It can also be used as an anticancer agent.

  Applicable cancer types when used as anticancer agents include, for example, stomach cancer, lung cancer, colon cancer, pancreatic cancer, liver cancer, ovarian cancer, breast cancer, prostate cancer, brain tumor, leukemia and the like.

  For preparations for use as pharmaceuticals, for example, injections, powders, granules, tablets, suppositories, capsules and the like can be applied, which are prepared by methods according to various conventionally known methods. It is used for pharmaceutical preparations as long as it does not adversely affect the pharmacological activity of bruceanol A reduced form, bruceanol B reduced form, bruceantinoside B reduced form or ailanthone reduced form, or pharmacologically acceptable salts thereof. Various pharmaceutical additives, that is, carriers and other auxiliaries such as stabilizers, preservatives, soothing agents, emulsifiers and the like may be used. In the preparation, the content of bruceanol A reduced form, bruceanol B reduced form, bruceantinoside B reduced form or ailanthone reduced form, or pharmacologically acceptable salt thereof can be changed widely depending on the dosage form and the like. In general, it is about 0.01 to 100% by weight, preferably about 0.1 to 70% by weight.

  As a method for administering the medicament of the present invention, various conventionally known methods such as injection, oral or rectal administration can be applied. The dose varies depending on the indication, symptoms, administration method, age, etc., but is about 0.01 to 800 mg per day per adult.

  The physiological activity of bruceanol A reductant, bruceanol B reductant, bruceantinoside B reductant, or ailanthone reductant will be described below.

Test Example 1 Cancer Cell Growth Inhibitory Action Human colon cancer cell HCT116, human kidney cancer cell A498, human lung cancer cell NCI-H460 using RPMI 1640 medium (manufactured by Iwaki) supplemented with 10% fetal bovine serum (manufactured by Moregate) Alternatively, human ovarian cancer cell OVCAR-3 was cultured at 37 ° C. under 5% CO ₂ . These cells were seeded in a 96-well plate and cultured for 1 day, after which a quasinoid compound was added. As a control, cisplatin used clinically was used.
After further culturing for 3 days, the cells were fixed with methanol and stained with methylene blue dye. After staining, the dye was extracted with 0.3% aqueous hydrochloric acid, the absorbance at 660 nm was measured, and the growth inhibitory activity was determined using the dose-response curve from the rate of decrease in the absorbance of the compound addition group relative to the absorbance of the cells to which no compound was added. The (IC ₅₀ ) value was determined and shown in Table 1.

Table 1 IC ₅₀ values (μM)

Results As is apparent from the above results, each quasinoid reductant has a strong cell growth inhibitory action against many types of human cancer cells.

Test Example 2 Adriamycin-resistant cancer cell growth inhibitory effect Using RPMI 1640 medium supplemented with 10% fetal bovine serum, mouse leukemia-derived cancer cells P388 or adriamycin-resistant strain P388 / ADR were cultured at 37 ° C. under 5% CO ₂ . . The cells were seeded in a 96-well plate and cultured for 1 day, and then bruceanol A reduced form, bruceanol B reduced form or adriamycin was added.
After further culturing for 2 days, 0.16 mg / ml WST-1 and 3.3 μg / ml 1-methyl-5-methylphenylmethylsulfate were added to the culture and cultured for 4 hours. A value obtained by subtracting the absorbance at 660 nm from the absorbance at 450 nm was determined. For the growth inhibitory activity, IC ₅₀ values were determined using a dose-response curve from the rate of decrease in the absorbance of the compound addition group relative to the absorbance of the cells to which no compound was added, and the values are shown in Table 2.

Table 2 IC ₅₀ values (μM)

Results As is clear from the above results, the quassinoid compounds of the present invention also exhibited cell growth inhibitory activity almost equal to that of sensitive cells (P388) against multidrug resistant cancer cells (P388 / ADR). This indicates that the quasinoid compound of the present invention can be a useful therapeutic agent for multi-drug resistant cancer cells which are clinically problematic.

  Although the Example of this invention is shown below, this is only an illustration, Comprising: This invention is not limited, Various modifications are possible.

Reference Example Extraction and isolation of bruceanol A, bruceanol B, bruceantinoside B, ailanthone 1) Isolation of bruceanol A and bruceanol B Elution was carried out sequentially using a mixed solvent to obtain 28 fractions. Of these, Fr16 was again subjected to silica gel column chromatography and purified using a mixed solvent of ethyl acetate-diethyl ether (1: 1 v / v) to obtain 6.7 g of a gum-like substance. This was subjected to preparative HPLC, and separation and purification were repeated to obtain 210 mg of bruceanol A and 103 mg of bruceanol B.

2) Isolation of bruceantinoside B Chloroform extract of 325 g of Brucea antioxidantidera above was subjected to silica gel column chromatography and eluted sequentially using a mixed solvent of chloroform-methanol to obtain 28 fractions. Furthermore, elution was successively performed with a chloroform-methanol-water mixed solvent (50: 14: 3 volume ratio) to obtain 7 fractions. Among these, the Fr3-7 fractions were subjected to Sephadex LH20 column chromatography and eluted with methanol. Of this, 26.2 g from which the dye portion had been removed was separated and purified by preparative HPLC to obtain 27 mg of bruceantinoside B.

3) Isolation of ailanthone 33 kg of Shinju (Ailanthus altissima) bark was extracted with methanol to obtain 2538 g of a crude extract. The solvent was partitioned sequentially using hexane, chloroform, and butanol. The solvent of the chloroform partition fraction was distilled off, and the residue (221 g) was subjected to silica gel column chromatography. Elution with a mixed solvent of ethyl acetate-diethyl ether (1: 1 v / v) gave 27 fractions. Fr12 was further separated and purified using preparative HPLC to obtain 32.2 mg of ailanthone.

Example 1 Bruceanol A Reductant Bruceanol A (20.2 mg) isolated above was dissolved in 15 ml of methanol, 30 mg of Pd / C (5%) was added, and the mixture was stirred under a hydrogen atmosphere for 24 hours. After completion of the reaction, the catalyst was filtered off and the reaction solution was concentrated and then subjected to silica gel column chromatography. Elution with a mixed solvent of ethyl acetate-diethyl ether (1: 1 v / v) gave 12.1 mg of bruceanol A reductant.
1. Appearance: colorless amorphous material Molecular formula: C ₂₈ H ₃₂ O ₁₁
3. Molecular weight: 544 (M ⁺ = 544 measured by EI-MS method)
4). 4. Infrared absorption spectrum (KBr method): 3400 (OH), 1740 (C═O), 1720 (C═O) cm ⁻¹ showing maximums Hydrogen nuclear magnetic resonance spectrum (solvent; C ₅ D ₅ N, internal standard; TMS) δ (ppm)
0.84 (3H, d, J = 6 Hz), 1.38 (3H, s), 1.54 (1 H, d, J = 13 Hz), 1.67 (1 H, m, H-4). 90 (1H, brd, J = 2Hz), 2.15 (1H, d, J = 13Hz), 2.21 (1H, d, J = 13Hz), 2.52 (1H, d, J = 13Hz), 2.66 (1H, d, J = 5 Hz), 3.84 (3H, s), 3.85 (1 H, d, J = 7 Hz), 3.91 (1H, d, J = 7 Hz), 4. 16 (1H, brs), 4.97 (1H, d, J = 7 Hz), 5.20 (1H, brs), 5.22 (1H, brd, J = 5 Hz), 5.29 (1H, d, J = 5 Hz), 6.65 (1H, brd, J = 13 Hz)
6). Solubility: soluble in water, acetone, methanol

Example 2 Bruceanol B Reductant Bruceanol B (20.3 mg) isolated above was dissolved in 15 ml of methanol, 30 mg of Pd / C (5%) was added, and the mixture was stirred under a hydrogen atmosphere for 24 hours. After completion of the reaction, the catalyst was filtered off and the reaction solution was concentrated and subjected to silica gel column chromatography. Elution with a mixed solvent of ethyl acetate-diethyl ether (1: 1 v / v) gave 14.2 mg of bruceanol B reductant.
1. Appearance: colorless amorphous material Molecular formula: C ₂₇ H ₃₈ O ₁₁
3. Molecular weight: 538 (M ⁺ = 538 measured by EI-MS method)
4). 4. Infrared absorption spectrum (KBr method): 3400 (OH), 1740 (C═O), 1720 (C═O) cm ⁻¹ showing maximums Hydrogen nuclear magnetic resonance spectrum (solvent; C ₅ D ₅ N, internal standard; TMS) δ (ppm)
0.85 (3H, t, J = 7 Hz), 0.86 (3H, d, J = 6 Hz), 1.38 (3H, s), 1.56 (1H, d, J = 13 Hz). 63 (1H, m), 1.97 (1H, brd, J = 2 Hz), 2.18 (1H, d, J = 13 Hz), 2.20 (1H, d, J = 13 Hz), 2.50 ( 1H, d, J = 13 Hz), 2.75 (1H, d, J = 5 Hz), 3.60 (1H, d, J = 7 Hz), 3.85 (3H, s), 3.91 (1H, d, J = 7 Hz), 4.18 (1H, brs), 4.92 (1H, d, J = 7 Hz), 5.08 (1H, brs), 5.18 (1H, brd, J = 5 Hz) 5.25 (1H, d, J = 5 Hz), 6.62 (1H, brd, J = 13 Hz)
6) Solubility: Soluble in water, acetone and methanol

Example 3 Bruceatinide B reductant Bruceatinide B (20.2 mg) isolated above was dissolved in 15 ml of methanol, 30 mg of Pd / C (5%) was added, and the mixture was stirred under a hydrogen atmosphere for 24 hours. After completion of the reaction, the catalyst was filtered off, the reaction solution was concentrated, and then separated and purified by preparative HPLC (ODS column, methanol-water 1: 1 v / v) to obtain 5.1 mg of bruceinoside B reductant. .
1) Appearance: colorless amorphous material 2) Molecular formula: C ₃₄ H ₄₈ O ₁₆
3) Molecular weight: 712 (M ⁺ = 712 measured by FD-MS method)
4) Infrared absorption spectrum (KBr method): 3400 (OH), 1740 (C═O), 1720 (C═O) cm ⁻¹ showing maximums 5) Hydrogen nuclear magnetic resonance spectrum (solvent; C ₅ D ₅ N , Internal standard; TMS) δ (ppm)
0.99 (3H, d, J = 7Hz), 1.45 (3H, s), 1.71 (1H, m), 2.97 (1H, d, J = 4Hz), 3.30 (1H, d, J = 13 Hz), 3.76 (1H, d, J = 8 Hz), 3.84 (3H, s), 4.29 (1H, brs), 4.66 (1H, d, J = 8 Hz) 4.80 (1H, brd, J = 5 Hz), 5.10 (1H, d, J = 5 Hz), 5.10 (1H, d, J = 5 Hz), 6.33 (1H, brd, J = 13Hz)
6) Solubility: Soluble in water, acetone and methanol

Example 4 alanthone reductant The alanthone (16.6 mg) isolated above was dissolved in 15 ml of methanol, 28.7 mg of Pd / C (5%) was added, and the mixture was stirred under a hydrogen atmosphere for 24 hours. After completion of the reaction, the catalyst was filtered off and the reaction solution was concentrated and purified by silica gel column chromatography. Elution was performed with a mixed solvent of ethyl acetate-diethyl ether (1: 1 v / v) to obtain 8.6 mg of reduced alanthone.
1) Appearance: colorless amorphous material 2) Molecular formula: C ₂₀ H ₂₈ O ₇
3) Molecular weight: 380 (M ⁺ = 380 measured by EI-MS method)
4) Infrared absorption spectrum (KBr method): 3380 (OH), 1740 (C = O), maximum at 1720 (C = O) cm ^-1 5) Hydrogen nuclear magnetic resonance spectrum (solvent CDCl ₃ , internal standard; TMS) δ (ppm)
0.86 (3H, d, J = 7 Hz), 1.24 (3H, d), 1.57 (3H, s), 1.70 (1H, m), 1.88 (1H, brd, J = 2 Hz), 2.05 (1 H, dd, J = 14 & 13 Hz), 2.18 (1 H, d, J = 13 Hz), 2.22 (1 H, d, J = 14 Hz), 2.48 (1 H, d, J = 13 Hz), 2.83 (1H, dd, J = 13 & 5 Hz), 2.90 (1H, dd, J = 18 & 5 Hz), 3.53 (1H, s), 3.66 (1H, d, J = 8 Hz), 3.69 (1 H, dd, J = 18 & 13 Hz), 4.11 (1 H, d, J = 8 Hz), 4.46 (1 H, brs), 4.54 (1 H, brs), 4.64 (1H, brs)
6) Solubility: soluble in water, acetone, methanol

Claims (8)

Formula (I)

A bioactive substance bruceanol A reduced form represented by the following formula (II)

Or a pharmacologically acceptable salt thereof. Formula (III) below

Or a pharmacologically acceptable salt thereof. A pharmaceutical comprising the compound according to claim 1 or 2 or a pharmacologically acceptable salt thereof as an active ingredient. A cell growth inhibitor comprising the compound according to claim 1 or 2 or a pharmacologically acceptable salt thereof as an active ingredient. The anticancer agent which uses the compound of Claim 1 or 2 or its salt accept | permitted pharmacologically as an active ingredient. Formula (IV) below

The pharmaceutical which uses the bioactive substance alanthone reductant represented by this, or its pharmacologically acceptable salt as an active ingredient. A cell growth inhibitor comprising the compound according to claim 6 or a pharmacologically acceptable salt thereof as an active ingredient. An anticancer agent comprising the compound according to claim 6 or a pharmacologically acceptable salt thereof as an active ingredient.