JPH0143756B2 - - Google Patents

Description

[Detailed Description of the Invention] (A) Background and Summary of the Invention The present invention is directed to a new antitumor agent, more specifically, as an intermediate for producing a carboxylic acid ester of 15β-hydroxyylanthone represented by the following formula (). Useful new ylanthone compound, 15β-hydroxy ylanthone triacylate (formula below)
Regarding.

(In the formula, R is an acyl group or benzoyl group having 2 to 5 carbon atoms substituted with halogen or not,
R' represents an alkene residue having 4 to 17 carbon atoms. ) Various natural products and compounds derived from natural products have been tested for their antitumor effects, and a small number of these have been put into practical use as anticancer agents, but they all have advantages and disadvantages in terms of efficacy, toxicity, etc. However, there is still nothing that is satisfactory. The present inventors have discovered that Ailanthus altissima Swingle, Ailanthus altissima Swingle,
Iylanthone 11β, 20-yupoxy, a triterpene contained in the plant body of Simarubaceae)
1β, 11α, 12α-trihydroxypiclaser-3,
13 (21) Jiyun-2,16-Zion (CHEMISTRY LETTERS, PP.661-662
(1982), the following formula), and its various derivations. As a result of synthesizing the compound and conducting pharmacological tests, it was discovered that the carboxylic acid ester of 15β-hydroxyylanthone represented by the above formula has a remarkable antitumor effect. The compound () of the present invention relates to a novel ylanthone compound that is important as an intermediate for obtaining the compound ().

The compound () of the present invention is produced using the above-mentioned ylanthone () as a starting material, for example, through the following steps.

That is, ylanthone () extracted and isolated from C. chinensis by a conventional method is acylated using an acylating agent such as acetic anhydride, acetyl chloride, chloroacetyl chloride, or benzoyl chloride in a conventional manner to obtain 1,12,20 - Obtain triacyl compound (). Since this reaction essentially seeks to protect the free hydroxyl groups in the compound () from the effects of reagents in subsequent steps such as reduction, dehydration, and oxidation, all conventional acylating agents can be used for this purpose. However, there is no need to limit the type of acylating agent. However, it is generally advantageous to use customary acylating agents such as those exemplified above.

Next, the triacyl compound ( ) protected with an acyl group is treated with a selective reducing reagent such as sodium boron hydride to reduce only the ketone group at position 16 and convert it into a secondary alcohol ( ). When this alcohol is treated with a dehydrating agent such as phosphorus acid chloride or phosphorus pentoxide, △ ₁₅ compound () is obtained, which is further oxidized using N-methylmorpholine-N-oxide and osmium tetroxide to give 15, Finally, when the latter is oxidized, only the hydroxyl group at the 16th position is oxidized, resulting in 15β-ylanthone-triacylate (1,12,20-triacylaylanthone-15β-), which is the object of the present invention.
All) () is obtained.

(B) Production example (i) Ilanthone-triacetate (1, 12,
20-Triacetyl ylanthone) () Production of ylanthone () 87.4g (0.23M)
Dissolve in 870 ml of anhydrous pyridine and then add 1700 ml of acetic anhydride. After the reaction mixture was left at room temperature for 20 hours, the solvent was distilled off under reduced pressure to obtain a crystalline product. When this crystalline substance is recrystallized using methanol, the desired product ()108.6
g is obtained as colorless needle crystals. Yield 93
%. The melting point and other physicochemical data of this product () are consistent with known ylanthone triacetate, and it does not show a drop in melting point even when mixed with a standard specimen of the latter.

(ii) Production of 16-hydroxyylanthone-triacetate (1,12,20-triacetyl ylanthone-16-ol) () 15 g of ylanthone triacetate ()
(30mM) 1.5% tetrahydrofuran and 1.5%
Dissolved in a mixture of ethanol and
2.25g of sodium borohydride while cooling on ice
(60mM). After stirring under ice cooling for 2 hours,
After adding a saturated ammonium chloride aqueous solution and further adding water to dissolve the formed precipitate, the organic solvent was removed under reduced pressure. The operation of adding methylene chloride to the resulting aqueous solution (residual liquid) and extracting it was repeated five times, and the separated organic solvent layer was washed twice with water, then once with saturated saline, and finally with magnesium sulfate. After drying, the solvent was removed under reduced pressure to obtain 14.7 g of the above target compound () as a colorless powder. Yield 98%.

Crystal form: Amorphous Optical rotation: [α] ²⁴ _D +5.00 (c=0.14, CHCl ₃ ) Infrared absorption (KBr, cm ^-1 ): 3450, 1750, 1740, 1680 Mass spectrum (m/e): M ⁺ 504, 462, 444, 420, 402, 378, 360 High resolution mass spectrum: Actual value m/e:
504.2013 Calculated value (as C ₂₆ H ₃₂ O ₁₀ ) m/e:
504.1996 Elemental analysis: C H Actual value: 61.76 6.39 Calculated value: 61.89 6.39 (iii) Production of 15,16-dehydroylanthone-triacetate (1,12,20-triacetyl ylanthone-15-ene) () 1. 12,20-Triacetyl ylanthone-
16-ol (16-hydroxyylanthone-
Dissolve 3.48 g (6.9 mM) of triacetate in 35 ml of anhydrous pyridine, and add 2.1 g (12.9 mM) of phosphorus oxychloride. The reaction solution was heated under reflux for 5 minutes under an argon stream, then water was added to the reaction solution under ice cooling to stop the reaction, and the pyridine was removed under reduced pressure. The remaining oil is diluted with water, extracted five times with ethyl acetate, and the organic solvent layer is separated. The obtained organic solvent layer is washed 2' times with saturated saline, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure to obtain colorless crystals. When the crystals are recrystallized from ethanol, 2.01 g of the desired _Δ15 compound () is obtained as colorless crystals. Yield 60%.

Crystal form: Colorless needles Melting point: 186-187℃ Optical rotation: [α] ²⁵ _D −2.86 (c=0.21, CHCl ₃ ) Infrared absorption (KBr, cm ^-1 ): 1760, 1680 Ultraviolet absorption: ( λ) 206nm (ε=12000), 238nm (ε=11400) Proton NMR (CDCl ₃ δppm): 1.44, s
(3H, 10-CH ₃ ); 1.94, s (3H, 4-CH ₃ ); 2.04, s (9H,
-OAc×3); 3.24, d(1H, 5α-H), 3.44; t(1H,
14β-H); 3.68, s(1H, 9α-H); 3.62, d, J=12Hz 1H 4.64, d, J=12Hz 1H(-CH ₂ -OAc): 4.16, t(1H, 7β-H) ;4.66, dd, J=7Hz,
3Hz (1H, 15-H); 5, 15, s (1H, 12-H);
5.24, s (1H, 1-H) 6.41, dd J=3Hz, 7Hz (1H, 16-H).

Mass spectrum (m/e): M ⁺ 486, 444, 426, 402, 384, 360, 342 Elemental analysis: C H Actual value 64.08 6.20 Calculated value 64.19 6.22 (iv) 15,16-dihydroxyylanthone-triacetate (1 , 12,20-triacetyl ylanthone-15,16-diol) () First, prepare a solution of the oxidizing agent using the following method. N-methylmorpholine-N-oxide
5.06g (43mM) in 72.5ml of water and 29.0ml of acetone
Dissolve in ml of mixed liquid and add 21.8 mg to this.
Add a solution of (0.0857 mM) osmium tetroxide in 10 ml of t-butanol.

Add 13.45 g (27.6 mM) of 1,12,20-triacetyl ylanthone-15 to the above oxidant solution.
-ene (15,16-dehydroylanthone-triacetate) () was added to the reaction solution to 145 ml.
Add acetone to dilute. After stirring at room temperature for 24 hours, add 290% sodium bisulfite to the reaction solution.
mg (2.79mM) and then add water to make a homogeneous solution. Concentrate this under reduced pressure to remove the solvent, and add 2N hydrochloric acid to the residue to lower the pH to 2. After adding salt to this to saturate the water, it was extracted 5 times with ethyl acetate, each extract (organic solvent layer) was combined, and after washing with saturated brine,
Dry with anhydrous sodium sulfite. When the solvent is removed from this dry organic solvent layer, the desired diol compound () is obtained as a colorless powder (this compound cannot be crystallized because it is a mixture of isomers regarding the 16-position hydroxyl group).

The above powder was coated with Mallinckrodt's silica gel (CC-7).
Apply 25g to column chromatography and elute with ethyl acetate to obtain 13g of pure product. Yield 90%.

Crystal form: Amorphous Optical rotation: [α] ²⁵ _D +3.81 (c = 0.21, CHCl ₃ ) Infrared absorption (cm ^-1 ): 3450, 1760, 1740, 1680 Mass spectrum (m/e): M ⁺ 520, 502, 478, 460, 436, 418, 400, 376, 358 High resolution mass spectrum: Actual value m/
e520, 1958; Calculated value (as C ₂₆ H ₃₂ O ₁₁ ) m/
e520.1943 Elemental analysis: C H Actual value: 59.82 6.21 Calculated value: 59.99 6.20 (v) Production of 15β-hydroxyylanthone-triacetate (1,12,20-triacetoxyylanthone-15β-ol) () Diol compound ()17.4g (33.4mM)
was dissolved in 850 ml of anhydrous acetonitrile, and 52.5 g (421 mM) of activated silver oxide was added thereto. After the mixture is heated and refluxed for 1 hour, it is allowed to cool to room temperature and the silver salt is removed using Celite. The solvent is removed from the solution under reduced pressure, leaving 22.6 g of oil. This oily substance was subjected to chromatographic purification using 300 g of the above-mentioned silica gel (CC-7), and the eluted portion with a mixture of ethyl acetate and chloroform = 4:6 was collected.
After concentrating the eluate, recrystallization from acetone-ether yields the final target compound 1, 12, 20-
10.4 g of triacetyl ylanthone-15β-ol (15β-hydroxyylanthone-triacetate) () are obtained as crystals.
Yield 60% (total yield from compound () 31.7
%). When the hydroxyl group at the 15th position of this product (the same applies to other acylated homologs) is changed to an unsaturated carboxylic acid ester, and the acetyl group is removed by further hydrolysis, the compound shown in the above formula () is obtained. This substance has a remarkable growth-inhibiting effect on tumor cells, and is expected to be effective as a new anticancer drug in the future.

Melting point: 231° to 232°C Crystal form: Colorless needle crystals Optical rotation: [α] ²⁵ _D +20.5 (c = 0.20, CHCl ₃ ) Infrared absorption (KBr, cm ⁺¹ ): 3540, 1760,
1740, 1680 Ultraviolet absorption λ ^EtoH _MAX : 238nm (ε=9000),
203nm (ε=5400) Proton NMR (CDCl ₃ , δppm): 1.41, s
(3H, 10-CH ₃ ); 1.95, s (3H, 4-CH ₃ ); 2.03 2.08 2.10s (3H, O ‖ -OC-CH ₃ ); 3.01, brd (1H, 5-H); 3.14, d, J=12Hz
(1H, 14β-H); 3.18, d, J = 2Hz (1H, 15β
-OH); 3.56, s (1H, 9α-H); 3.56, d, J = 12Hz (1H) 4.52, d, J = 12Hz (1H) - CH ₂ - OAc; 4.69, h (1H, 7β-H ); 5.16, s (1H, 12-
H); 5.04, dd, J=2Hz, 12HZ (1H, 15α-H); 6.04, q (1H, 3-H) Mass spectrum (m/e): M ⁺ 518, 476, 458,
434, 416, 392, 374 Elemental analysis: C H Actual value: 60.27 5.80 Calculated value: 60.23 5.86 X-ray analysis: R value 7.1% (excluding |F _OBS | = 0) 0.071 Crystal Recrystallized with ethanol and water Specific gravity 1.37 Space group p2, (single needle) Z=2 Lattice constant a=10.254A (Q(a)=0.001) b=14.560A (Q(b)=0.002) c=8.4670A (Q(c)=0.0009) β=101.450 (Q(β)=0.01) (C) Reference example (i) General production of 15β-hydroxyylanthone-triacetate unsaturated carboxylic acid esters Unsaturated carboxylic acid (1.24 equivalent), 1-ethyl-2- Fluoropyridinium tetrafluoroborate or 1-methyl-2-fluoropyridinium tosylate (1.50 equivalents) is dissolved in anhydrous methylene chloride, and cesium fluoride (4.87 equivalents) is added thereto. The solution was stirred for 30 min at room temperature and 1.00 g of 15β-hydroxyylanthone-triacetate () was added under a stream of _N2 .
(1.93mM) all at once. then at room temperature
After stirring for 20 hours, water is added to the reaction solution to terminate the reaction. The reaction mixture is extracted three times with methylene chloride, and the resulting solvent layer is washed twice with saturated sodium bicarbonate, then twice with saturated brine, and then dried over anhydrous magnesium sulfate. This solvent layer is concentrated under reduced pressure to leave an oil. After adsorbing this residue on a column packed with 30 g of the above-mentioned silica gel (cc-7), benzene:ethyl acetate=
Elution with a 9:1 mixture yields the title compound as a colorless oil.

(ii) Production of 15β-hydroxy ylanthone-unsaturated carboxylic acid ester () 1.0 mM of the triacetyl ylanthone-carboxylic acid ester obtained in Reference Example (i) was added to a methanol solution containing 0.02 equivalents of potassium methoxide. Dissolve and stir at room temperature for 2 hours under _N2 . Add 1N hydrochloric acid to the reaction solution to adjust the pH to 4-5.
After methanol is distilled off, water is added to the residue and extracted three times with methylene chloride. The separated methylene chloride layer is washed twice with saturated saline and dried using anhydrous magnesium sulfate. The solvent was removed from this dried extract under reduced pressure, and the residue was transferred to a silica gel column (Merck Kieselgel 60, 70-230 mesh ASTM).
After loading, ethyl acetate:hexane=5:
When eluted with a mixture of 2 and 2, the above compound () is obtained. This one is a mouse (BDF ₁ ,
In animal experiments using male mice, 4-5 weeks old, average weight 18g), it has a remarkable survival effect on murine leukemia P388 cell infection.

The mixture of monoacetate and diacetate separated by the above chromatography was stirred for 48 hours at room temperature in a mixture of 10 times the amount of anhydrous pyridine and 20 times the amount of clear acetic acid. After acetylation, the target compound can be derived by hydrolyzing it in the same manner as above.

Claims (1)

[Claims] 1. General formula (In the formula, R is an acyl group having 2 to 5 carbon atoms or a benzoyl group, which may or may not be substituted with halogen). 2 formulas 2. The compound according to claim 1, represented by: