JPH0552292B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION A. Field of Industrial Application of the Invention The present invention relates to an antitumor agent containing achimylic acid A lactone represented by the formula ().

Conventional technology As an antitumor higher plant component, tripdiolide [J.Amer.Chem.Soc., 94 ,
7194, (1972)], bruceantin
There have been many reports such as [J.Org.Chem., 38 , 178, (1973)], but the ones that have been put into practical use are the vinca alkaloid vinblastine [The Merck Index 10th edition 9784],
Examples include vincristine [Biologically Active Natural Substances 431 (1978)]. As structurally similar substances, stereoisomers of seco-tanapartholide A and B (seco-tanapartholide AandB)
There are examples such as [Phytochemistry 21 , 2543 (1982)], but no antitumor effects have been reported.

Problems to be Solved by the Invention Various natural products and compounds derived from natural products have been tested for their antitumor effects, and some alkaloids and protein-bound polysaccharides have not been put to practical use as antitumor agents. However, natural polyoxides have many problems in terms of efficacy and toxicity, and none have yet been put to practical use.

B. Means for Solving the Constituent Problems of the Invention The present inventors have been conducting research on natural products that have antitumor effects. As a result, Achillea millefolium L,
We discovered an active substance with antitumor effects in polyoxides contained in plants of the Compositae and named it achimylic acid. Achimylic acid A lactone represented by the formula () can be isolated from Yarrow as follows. Dried yarrow flowers are extracted with a polar organic solvent at room temperature for several days, and then the extract is separated with water using a lower alkyl alcohol that is immiscible with water, such as n-butanol or pentanol. To obtain acidic substances, first extraction is performed with a basic solution, and then the aqueous layer is made acidic and extracted using a suitable organic solvent that is immiscible with water. Further, by separating the obtained acidic substance using silica gel column chromatography, an active substance against P388 can be obtained. Examples of polar organic solvents include methanol, ethanol, isopropanol,
n-butanol, sec-butanol or tret-
Examples include lower alcohols such as butanol, acetone, and acetonitrile. As the basic solution, a moderate base such as a saturated aqueous solution of sodium bicarbonate, sodium carbonate or sodium acetate is preferred. Organic solvents that are immiscible with water include dichloromethane, ethyl acetate, n-
Examples include butanol or pentanol. To obtain achimylic acid A lactone from the active substance, the active substance is dissolved in a suitable solvent such as chloroform, dichloromethane or methanol and left at room temperature for several days. When the solvent is distilled off and the residue is developed and eluted using silica gel column chromatography, a crude lactone can be obtained. The crude lactone is separated by silica gel column chromatography for further purification. As the elution solvent, for example, chloroform, dichloromethane, ether, ethyl acetate or benzene may be used alone or in combination depending on the polarity of the substance to be eluted.

The antitumor agent of the present invention may be formulated according to conventional methods. Specifically, it is produced by using a compound represented by the formula () as an active ingredient, mixing this with a suitable base, and then shaping the mixture as necessary.
When the antitumor agent is an injection, it is manufactured by sterilization together with an appropriate carrier. The formulation is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and lubricants. Typical formulations include tablets, pills, powders, solutions, suspensions,
Examples include emulsions, granules, capsules, suppositories, injections (solutions, suspensions, etc.). For example, lactose,
Excipients such as white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid; Binders such as water, ethanol, propanol, simple syrup, glucose solution, starch solution; dried starch, alginic acid Disintegrants such as sodium, agar powder, sodium bicarbonate, calcium carbonate, and sodium lauryl sulfate; disintegration inhibitors such as sucrose and stearin; humectants such as glycerin and starch; lubricants such as purified talc, stearate, and boric acid powder; It may be used with additives known in the art such as brighteners. In addition, when preparing an injection such as a solution, emulsion, or suspension, diluents such as water, aqueous lactic acid solution, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, and polyoxylated isostearyl alcohol may be used. , polyoxyethylene sorbitan fatty acid esters, etc. may be used, and if desired, salt, glucose, or glycerin may be added to obtain isotonicity, and conventional solubilizing agents, buffers, soothing agents, etc. may be added. Further additions may also be made.

The dosage of the preparation of the present invention varies depending on the usage, patient's age, sex, degree of disease, etc., and cannot be unconditionally prescribed.
The amount of lactone is usually 100 to 2000 mg per day for adults orally, and 10 to 200 mg per day parenterally.
may be administered in 1 to 3 divided doses.

Examples and physical constants are shown below to clarify aspects of the present invention, but the scope of the present invention is not limited thereto.

Examples Example 1 Extraction of active substances from the flowers of Yarrow yarrow Dried Yarrow (scientific name:
Achillea millefolium L.) flowers were soaked in methanol (500 ml) for 3 days at room temperature for extraction. This operation was repeated three times, and methanol was distilled off under reduced pressure to obtain 9.6 g of methanol extract as a brown oil. This methanol extract is divided between n-butanol (100 ml) and water (50 ml), and the n-butanol layer is extracted with 5% sodium hydrogen carbonate solution (20 ml x 6). Add dilute hydrochloric acid to the alkaline solution layer to adjust the solution to pH 3, and extract with ethyl acetate (20 ml x 5). After washing the solvent layer with water and drying, the solvent was distilled off under reduced pressure to obtain 324 mg of the acidic substance. This acidic substance is subjected to adsorption chromatography using 20 g of Merck's silica gel (0.063-0.200 mm). Chloroform-methanol-water (85:
15:1.5) 80ml followed by the same solvent (80:20:2)
When the solution is developed and eluted with 30 ml, 70 mg of active substance against P388 is obtained as a pale yellow oil from the fraction eluted in the latter half.

Example 2 Achimylic acid A lactone 200 mg of active substance is dissolved in 2 ml of chloroform,
After standing at room temperature for 2 days, the solvent was distilled off. residue
200 mg is chromatographed using 15 g of silanized silica gel (0.063-0.200 mm) manufactured by Merck. Methylene chloride-n-hexane (4:1) 80ml
After development, 142 mg of the crude lactone compound was obtained by elution with 50 ml of the same solvent (9:1). The crude product was again subjected to chromatography using 15 g of silanized silica gel (same as above) as a carrier, developed and eluted with benzene-ethyl acetate (95:5). 107 mg of achimylic acid A lactone are obtained as a colorless oil. [α] _D 6.1゜ (CHCl ₃ ), IR: νmax (film) 3440, 1765, 1710,
1660, 1593, 1370, 1270, 1240, 1120, 940, 810
cm ^-1 , MS: 278 [M ⁺ ], ¹³ CNMR: δppm (CDCl ₃ )
208.8, 202.8, 170.2, 167.0, 137.7, 130.9,
124.9, 80.5, 78.7, 62.6, 42.1, 39.7, 30.0,
28.6, 25.0.

C. Effects of the Invention We conducted an activity test of the compounds handled in the present invention against P388 leukemia mice. Six or seven 5- or 6-week-old BDF1 mice (male) were used for each administration as test animals. P388 leukemia was used as the test cancer type, and 10 ⁶ cancer cells were intraperitoneally administered to each test animal. The specimens were administered as suspensions at predetermined doses one day after cancer transplantation, or for 5 days in the case of continuous administration. In the control group, only the suspension solvent was administered intraperitoneally. ILS% at each dose after study completion
(Increased life spen%, life extension rate) was calculated.
ILS of 30% or higher was judged to be effective.

ILS=S _T −S _C /S _C ×100 S _T : Days of survival of experimental animals S _C : Number of days of survival of control animals When achimylate A lactone was administered once at 2 mg/Kg and 5 mg/Kg, respectively, the ILS was 38%,
When 1 mg/Kg and 4 mg/Kg were administered continuously, ILS was 48% and 40%, respectively. The above results prove that achimylic acid A lactone has a remarkable antitumor effect.

Claims (1)

[Claims] 1 Formula: An antitumor agent containing achimylic acid A lactone represented by: