JPH0578536B2 - - Google Patents

Description

[Detailed description of the invention]

(Technical Field) The present invention relates to a novel dihydrochalcone compound and an anticancer agent containing the compound as an active ingredient. (Background of the Invention) Conventionally, alkylating agents (nitrodiene mustards, ethyleneimines,
sulfonic acid esters), antimetabolites (folate antagonists, purine antagonists, pyrimidine antagonists), plant fission toxins (colcemid, pinblastin, etc.),
Antibiotics (galcomycin, carcinophilin,
mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and porphyrin complex salts (marphyrin, COPP), etc. are used. However, most of them are cytotoxic substances and exhibit serious side effects, so there is a strong desire to develop anticancer agents with low toxicity and excellent anticancer activity. In view of the above purpose, the present inventors have been searching for substances with low toxicity and anticancer properties from a wide range of organisms in the animal, plant, and microbial worlds.
The present invention was completed based on the discovery that a novel dihydrochalcone compound extracted from Artocarpus communis (Indonesian name: Kulur) has excellent anticancer activity. Artocarpus commienis is a plant that grows wild in various parts of Indonesia, and its cluster flowers are said to have been used as a substitute for mosquito coils since ancient times. However, there have been no reports on research on the components of this plant. This time, the inventors
As a result of exploring the physiological activity of the components of the petals of Artocarpus comienis, we obtained the novel dihydrochalcone compound of the present invention, and obtained the novel finding that this compound exhibits excellent anticancer activity. (Objective of the Invention) An object of the present invention is to extract and isolate a novel dihydrochalcone compound having more anticancer activity. Another object of the present invention is to provide an anticancer agent containing the above novel dihydrochalcone compound as an active ingredient. (Structure of the invention) The novel dihydrochalcone compound of the present invention, 2-
Geranyl-3,4,2',4'-tetrahydroxychalcone has the physical properties described below and is extracted and isolated from Artocarpus commienis. below,
An example of the extraction/isolation method is shown below. The flower parts of Artocarpus communis collected on Jiyawa Island, Indonesia are extracted several times with 90% ethanol solution and concentrated under reduced pressure to obtain a crude extract. Next, the obtained crude extract is subjected to adsorption chromatography and sequentially eluted with aqueous methanol, methanol, and acetone. The methanol elution fraction and the acetone elution fraction are combined and concentrated under reduced pressure to obtain a brown oil. This product exhibits a significant growth inhibitory effect on cultured Yoshida sarcoma cells. Therefore, the above oil was further subjected to silica gel column chromatography (solvent: hexane/ethyl acetate = 10:1 to 0:100), and 11 fractions (AC1 to
11). Each of the obtained fractions was examined for its growth inhibitory effect on cultured Yoshida sarcoma cells. Next, among these fractions, AC-
The fractions of 5 and AC-6 were collected and further subjected to high performance liquid chromatography (solvent: hexane/ethyl acetate =
Separation at a ratio of 2:1) yields the novel dihydrochalcone compounds of the present invention. The dihydrochalcone compound of the present invention has the following structural formula and physical properties.

[Physical properties]

MS: m/e410 (M ⁺ ), C ₂₅ H ₃₀ O ₅ IR (KBr): 3450 (OH), 1620 (hydrogen-bonded C=
O) cm ^-11 H-NMR (90MHz, CDCl ₃ , δ):
1.56 (3H, bs), 1.65 (3H, bs), 1.78 (3H, bs),
2.04 (4H, bs), 2.80−3.20 (4H, m), 3.40
(2H, d, J=6.6Hz), 4.88−5.30 (2H, m),
6.32 (1H, dd, J=2.4, 9.5Hz), 6.37 (1H,
bs), 6.62, 6.73 (2H, ABq, JP9.2Hz) ¹³ C−
NMR (22.5MHz, CDCl ₃ , ε): 16.1(q), 17.4
(q), 25.4(q), 26.3(t), 27.7(t), 39.4(two
carbon, t), 58.6(t), 103.4(d), 108.1(d),
112.9(d), 113.3(s), 121.0(d), 121.8(d), 123.7(d),
126.3(s), 131.3(s), 131.7(s), 132.2(d), 137.7(s),
142.2(s), 142.4(s), 163.2(S), 164.8(s), 204.3(s) This compound shows no toxicity to mice even after continuous administration of 50mg/Kg. The anticancer agent of the present invention can be administered either orally or parenterally.
Administered as hard capsules or tablets, granules, fine granules, or powders, and for parenteral administration, subcutaneous or intravenous injections such as aqueous suspensions and oil-based preparations, infusions, and solid or suspended viscosity. It can be administered in the form of a suppository so that sustained mucosal absorption can be maintained in the form of a thick liquid. The formulation of the active ingredient of the present invention includes surfactants, excipients, lubricants, adjuvants, and, if necessary, pharmaceutically acceptable film-forming substances and coating aids to form enteric-coated formulations. Specific examples thereof are as follows. In order to improve disintegration and elution of the composition of the present invention, one or two surfactants such as alcohols, esters, polyethylene glycol derivatives, sorbitan fatty acid esters, sulfated fatty alcohols, etc. are used. The above can be added. In addition, excipients such as sucrose, lactose, starch, crystalline cellulose, mannitol, light silicic anhydride, magnesium aluminate, magnesium aluminate metasilicate, synthetic aluminum silicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, carboxylic Methylcellulose calcium and the like can be added alone or in combination of two or more. As a lubricant, for example, one or more types of magnesium stearate, talc, hydrogenated oil, etc. can be added, and as a flavoring agent and a flavoring agent,
Sweeteners such as salt, saccharin, sugar, mannitrite, orange oil licorice extract, citric acid, glucose, menthol, eucalyptus oil, and malic acid, fragrances, colorants, preservatives, and the like may be included. Adjuvants such as suspending agents and lubricants may include, for example, coconut oil, olive oil, sesame oil, peanut oil, potassium lactate, safflower oil, soybean phospholipid, and the like. Film-forming substances include cellulose acetate phthalate (CPA) as a carbohydrate derivative such as cellulose and sugars, and methyl acrylate and methacrylate as polyvinyl derivatives such as acrylic acid copolymers and dibasic acid monoesters. Examples include acid copolymers and methyl methacrylate/methacrylic acid copolymers. In addition, when coating the above-mentioned film-forming substance, in addition to commonly used coating aids such as plasticizers, various additives to prevent chemicals from adhering to each other during coating operations can be added to the film-forming agent. properties and make coating operations easier. In addition,
It is also possible to form a dosage form in which the active ingredient is microprocapsulated using a film-forming substance and then mixed with excipients and the like. Next, the compounding ratio in typical dosage forms is as follows. Particularly preferred range Active ingredients 0.1-90% by weight 0.3-15% by weight Excipients 10-99.8 〃 85-99.4 〃 Lubricants 0-50 〃 0-20 〃 Surfactants 0-50 〃 0-20 〃 Film formation Substance 0.1-50 0.3-20 Particularly preferred excipients are lactose, crystalline cellulose, calcium carboxymethyl cellulose. The dosage is sufficient to effectively treat the target tumor, and depends on the symptoms of the tumor, route of administration, dosage form, etc., but in general, when administered orally, the amount per day for adults is , in the range of about 0.01 to 100 mg/Kg body weight (0.01 to 60 mg/Kg body weight for children), and the upper limit is preferably about 50 mg/Kg body weight, more preferably about 10 mg/Kg body weight, and the upper limit is about 10 mg/Kg body weight. In this case, the upper limit is approximately 10 mg/Kg body weight, preferably 5 mg/Kg body weight, and more preferably 2 mg/Kg body weight. Next, the anticancer activity test method for confirming the anticancer activity of the compound of the present invention will be described. Yoshida sarcoma cells taken from rat ascites were inoculated into MEM medium (manufactured by Nitsusui) containing 15% calf serum and cultured.
When the cell number reaches approximately 10 x 10 ⁵ cells/ml, dilute it 5 times with the above medium (20 x 10 ⁴ cells/ml) and add 1 ml.
Dispense into vials. Next, a methanol or acetone solution of the compound of the present invention at each concentration is added, a rubber stopper is placed, and the cells are cultured at 37°C. After about 3 days, the number of living cells is counted after staining with trypan blue. The inhibition rate of test cell proliferation was determined by the following formula. Inhibition rate (%) = (1 - number of living cells in vial administered with test compound/number of living cells in vial not administered) x 100 The present invention will be specifically described below with reference to Examples, Formulation Examples, and Test Examples. explain. Example: Approximately 500 g of the flower part of the Indonesian medicinal plant Artocarpus commienis was extracted several times with 90% ethanol, and the extract was concentrated under reduced pressure to obtain approximately 20 g of an oily substance.
get. Next, this oily substance was mixed with DiaionHP-20
It was subjected to column chromatography packed with 40
Elute sequentially with % hydrated methanol, 60% hydrated methanol, 80% hydrated methanol, methanol, and acetone. Next, the methanol elution fraction and the acetone elution fraction were combined and concentrated under reduced pressure to obtain 15.7 g of a brown oil. This product shows a remarkable inhibitory effect on cultured Yoshida sarcoma cells. Next, the above oily substance was further subjected to silica gel column chromatography (solvent: hexane/ethyl acetate = 10:1 to 0:100, methanol), and 11
fraction (AC-1: 2.0g, AC-2: 0.6g, AC
-3: 0.8g, AC-4: 0.3g, AC-5: 1.0g,
AC-6: 3.0g, AC-7: 0.3g, AC-8: 2.4
g, AC-9: 0.2g, AC-10: 3.0g, AC-
11:1.0g). For each fraction obtained,
We investigated the growth inhibitory effect on cultured Yoshida sarcoma cells.
Among these fractions, AC-
Fractionation of 5 and AC-6 (hexane:ethyl acetate=
5:2) was further fractionated using high performance liquid chromatography (Nucleosil 50-5, solvent: hexane:ethyl acetate = 2:1, flow rate 9 ml/min), and the retention time
A peak around 28 minutes was collected and concentrated to obtain 2-geranyl-3,4,2',4'-tetrahydroxychalcone (950 mg). Formulation Example 1 (Injection/Drop) Add 5 g of powdered glucose to contain 10 mg of the compound of the present invention, dispense aseptically into vials, seal them, fill with inert gas such as nitrogen or helium, and store in a cool, dark place. saved. Before use, dissolve in ethanol and add 100 ml of 0.85% physiological saline to make an intravenous injection, and administer 10 to 100 ml per day by intravenous injection or drip depending on the symptoms. Formulation Example 2 (Injection/Drop) Using 2 mg of the compound of the present invention, an intravenous injection for mild symptoms was prepared in the same manner as in Formulation Example 1, and administered for 10 to 10 days.
Administer 100ml by intravenous injection or drip depending on symptoms. Formulation Example 3 (Enteric Capsules) 5 g of the compound of the present invention, 2.46 g of lactose and 0.04 g of hydroxypropylcellulose were each taken and mixed well, then formed into granules according to a conventional method, dried thoroughly and sieved. We have manufactured granules suitable for separate bottles, heat seal packaging, etc. Next, 0.5 g of cellulose acetate phthalate and 0.5 g of hydroxypropyl cellulose phthalate were dissolved to form a coating base material, and the granules were coated with this base material while floating and flowing to obtain an enteric granule material. This composition is filled into capsules to produce 100 enteric capsule formulations. Test Example (Anticancer Activity Test) When the growth inhibition rate (%) of Yoshida sarcoma cells was calculated using the compound of the present invention according to the test method described above, the results shown in Table 1 were obtained.

【table】

Claims (1)

[Claims] 1. A dihydrochalcone compound represented by the structural formula: [Claims] 2 Structural formula: An anticancer agent containing a dihydrochalcone compound represented by the following formula as an active ingredient.