JPH0310631B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel diterpenoids. The present inventors have been conducting repeated research on extracts from Kurobana Hikiokoshi, a plant belonging to the Lamiaceae family, genus Yamahathuka, grown in Hyogo Prefecture, and have already filed a patent application for the trichorrhabdals obtained by the extraction. 56-129840 (see JP-A-58-32874)).
The present inventors have succeeded in synthesizing a new diterpenoid with even more excellent antitumor activity by further subjecting these tricholabdals to acetylation, catalytic reduction, hydrolysis, etc., and have completed the present invention. . The novel diterpenoids of the present invention are represented by the general formula: (In the formula, R ¹ means a methyl group or a methylene group, R ² means a hydrogen or hydroxyl group, and R ³ means a hydrogen or acetoxy group.) The tricholabdals used as the raw material of the present invention are components of black-and-white plant. Kurobana Hikiokoshi belongs to the Lamiaceae family and is also called Enmeisou.It is a perennial plant that grows naturally in the mountains and has been used in folk medicine since ancient times as a bitter-tasting stomachic herb, and is said to be especially effective for abdominal pain. The compound of the present invention is a novel tricholabdal derivative obtained by subjecting these extracted components tricholabdals to reactions such as acetylation, catalytic reduction, and hydrolysis, and has excellent antitumor effects. A representative example of the diterpenoid of the present invention is specifically represented by the following structural formula. As mentioned above, the diterpenoids of the present invention can be obtained, for example, by subjecting tricholabdal B, tricholabdal C, etc. synthesized in Japanese Patent Application No. 129840/1984 to reactions such as acetylation, catalytic reduction, and hydrolysis. Here, the raw material tricholabdar B or C has the following chemical structural formula, and these are obtained by drying the leaves of the black-and-white plant and using an organic solvent such as ether, ethyl acetate, acetonitrile, acetone, methanol, ethanol, dichloride methane, or chloroform. After extraction under reflux, the concentrated residue is washed with water to remove sugars, and then subjected to column chromatography. The above acetylation is carried out by reacting acetic anhydride, acetyl chloride, etc. in the presence of a catalyst or in the absence of a catalyst. As the catalyst, commonly used pyridine, triethylamine, dimethylaminopyridine, etc. are used, and as the solvent, although not particularly limited as long as it does not participate in the reaction, pyridine, chloroform, dichloromethane, etc. are generally used. Catalytic reduction is carried out by adding hydrogen in the presence of a catalyst such as palladium on carbon, Raney nickel, or platinum. The solvent is not particularly limited as long as it does not participate in the reaction, but acetic acid, ethyl acetate, ethanol, methanol, etc. are generally used. For hydrolysis, commonly used acids such as dilute hydrochloric acid and dilute sulfuric acid are used in a conventional manner. The solvent is not particularly limited as long as it does not participate in the reaction, but water-soluble solvents such as dimethylformamide, tetrahydrofuran, and dioxane are generally used. When the compound of the present invention thus obtained is used as a medicine, it can be formulated into a formulation depending on the route of administration, together with a conventional pharmaceutical carrier. For example, for oral administration, tablets, capsules, granules, powders,
For parenteral administration, it is prepared in the form of an injection, a suppository, etc. As carriers that can be used in preparing solid preparations for oral administration, conventional excipients, binders, lubricants, coloring agents, disintegrants, etc. can be used. Examples of excipients include lactose, sucrose,
Starch, talc, magnesium stearate,
Crystalline cellulose, methyl cellulose, carboxymethyl cellulose, glycerin, sodium alginate, gum arabic, etc. are used as binders, polyvinyl alcohol, polyvinyl ether, ethyl cellulose, gum arabic, citric acid, white sugar, etc. are used as lubricants, magnesium stearate,
In addition to talc and other coloring agents and disintegrants, commonly known ones can be used. Furthermore, the tablets may be coated by a well-known method. Liquid preparations may be aqueous or oily suspensions, solutions, syrups, elixirs, etc., and are prepared by commonly used methods. When preparing an injection, the compound of the present invention is added with a PH adjusting agent, a buffering agent, a stabilizer, an isotonic agent, and a local disintegrating agent, and the injection is prepared for subcutaneous, intramuscular, or intravenous use using a conventional method. can be manufactured.
Bases for producing suppositories include, for example, cocoa butter, polyethylene glycol, lanolin, fatty acid triglycerides, Huitepsol (registered trademark),
An oil-based base such as Dynamite Nobel Co., Ltd.) can be used. The dosage of the preparation thus prepared varies depending on the patient's symptoms, body weight, age, etc., and cannot be absolutely limited, but it is usually in the range of about 50 to 1000 mg of the compound of the present invention per day for adults. This is usually preferably administered in 1 to 4 divided doses per day. Next, the production method, physical properties, and pharmacological action of the tricholabdal derivative, which is the compound of the present invention, will be described in more detail with reference to Examples. Example 1 Tricolabdar B (621 mg) was dissolved in 30 ml of distilled tetrahydrofuran, 30 drops of 5% hydrochloric acid were added dropwise, and the mixture was heated under reflux for 7 hours under a nitrogen stream. Pour the reaction solution into 50 ml of ice water and dilute with ethyl acetate for 200 ml.
Each ml was extracted three times. After washing with saturated brine to make it neutral, it was dried over sodium sulfate, and the solvent was removed under reduced pressure. 620 mg of residue was obtained, which was separated by silica gel column chromatography to obtain 239 mg (43%) of hemiacetal product and 156 mg (25%) of raw material. After reacting 190 mg of this hemiacetal product with 4 ml of acetic anhydride and 4 ml of pyridine at room temperature overnight,
The solvent was removed under reduced pressure. The residue is chloroform-
Recrystallization was performed from 10 ml of a methanol mixture (1:1) to obtain 170 mg of the desired tricholabdal B derivative [compound ()] having a melting point of 280°C. Elemental analysis value (as C ₂₂ H ₂₈ O ₇ ) C H Theoretical value (%) 65.33 6.98 Actual value (%) 65.29 6.93 NMR spectrum (d ₅ -pyridine, TMS) δppm 1.00 (3H, s, -CH ₃ ) 2.19 (3H, s, -COCH ₃ ) 2.65 (1H, d, J=6Hz 5-H) 3.10 (1H, dd, J=10,4Hz, 13-
H) 3.46, 3.84 (each 1H, ABq, J=8Hz, 19−
H ₂ ) 3.63 (1H, d, J=11Hz, 14−Hα) 4.02 (1H, dd, J=12,3Hz, 20−
H _A ) 4.46 (1H, m, 11-H) 5.28 (1H, d, J=12Hz, 20-H _B ) 5.46, 6.12 (each 1H, br, s, exomethylene) 6.81 (1H, d, J=6Hz , 6-H) 6.88 (1H, d, J=4Hz, -OH,
IR spectrum ν _nax (KBr) ₃₅₄₀ , 1740, 1720, 1700, 1640,
1240cm ^-1 Example 2 After dissolving Tricolabdar B (1g) in 40ml of distilled tetrahydrofuran, 50 drops of 5% hydrochloric acid were added dropwise, and the mixture was heated under reflux for 8.5 hours under a nitrogen stream. Thereafter, 709 mg (79%) of hemiacetal product was obtained in the same manner as in Example 1. After reacting 423 mg of this with 8 ml of acetic anhydride and 8 ml of pyridine at room temperature overnight, the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate to obtain 200 mg of crystals. 200 mg of this acetylated product was dissolved in 20 ml of distilled methanol, 40 mg of palladium-carbon (10%) was added, and the mixture was brought into contact with hydrogen at room temperature overnight. After the reaction is complete,
Filter through paper (filter No. 6), concentrate the liquid, and recrystallize from 10 ml of a chloroform-methanol mixture (1:1) to obtain the desired tricholabdal B derivative [compound ()] with a melting point of 199-201°C. Obtained 153mg. Mass spectrum analysis results m/e = 406 (M ⁺ ) NMR spectrum (CDCl ₃ + d ₅ -pyridine,
TMS) δppm 1.00 (3H, s, 18−CH ₃ ) 1.13 (3H, d, J=7Hz, 17−CH ₃ ) 2.15 (3H, s, −COCH ₃ ) 2.26 (1H, d, J=5Hz, 5 −H) 2.00 (1H, dd, J=10,4Hz, 14−
Hβ) 2.50 (1H, q-d, J=7.7Hz,
16-H) 3.44, 3.81 (each 1H, ABq, J=8Hz, 19-
_H2 ) 3.79, 5.00 (each 1H, dd, J=12, 2Hz, 20
−H _A ) d, J=12Hz, 20−
H _B ) 3.52 (1H, d, J = 10Hz, 14-Hα) 4.31 (1H, m, 11-H) 5.73 (disappeared with 1H, -OH, D ₂ O) 6.40 (1H, d, J = 5Hz, 6-H) IR spectrum ν _nax (KBr) 3540, 3500-3420, 1745, 1730,
1695, 1230cm ^-1 Example 3 Tricholabdal C (320mg) was added to acetic acid (special grade) 5
ml and heated at 90°C for 10 hours under a nitrogen stream. After the reaction, acetic acid was removed under reduced pressure, and the residue was separated by silica gel column chromatography. Fraction 130mg of the target compound () and mixed fraction 69 with other components
Both mg were crystallized from ethyl acetate, melting point 218.5.
143 mg of the desired tricholabdal C derivative [compound (2)] was obtained at ~220.5°C. Mass spectrum analysis results m/e = 446 (M ⁺ ) NMR spectrum (d ₅ -pyridine, TMS) δppm 1.17 (3H, s, -CH ₃ ) 2.04 (3H, s, 3-OCOCH ₃ ) 2.18 (3H, s , 6-OCOCH ₃ ) 2.43 (1H, d, J=12Hz, 14-Hα) 2.64 (1H, dd, J=5, 1Hz, 5-
H) 2.96 (1H, dd, J=9.4Hz, 13-
H) 3.67, 3.80 (each 1H, ABq, J=8Hz, 19−
H ₂ ) 3.92 (1H, dd, J = 12, 2Hz, 20−
H _A ) 4.27 (1H, d, J=12Hz, 20−H _B ) 5.08 (1H, dd, J=11,4Hz, 3−
H) 5.43, 6.09 (each 1H, br, s, exomethylene) 6.78 (1H, d, J=5.2Hz, 6-H) IR spectrum ν _nax (KBr) 1740 (sh), 1730, 1720, 1700,
1638, 1245 cm ^-1 Pharmacological Test The anticancer activity of the trichorabdal B derivative [compound ()] of the present invention and tricholabdal B was compared. 2 x 10 ⁶ of Ehrlitsu carcinoma ascites cells
cells/mouse were implanted intraperitoneally into male ddY mice (25-28 g). Compound () or tricholabdar B was dissolved or suspended in physiological saline and intraperitoneally administered to 7 mice per group at a volume ratio of 0.1 ml/10 g mouse body weight once a day for 7 consecutive days starting the day after tumor implantation. . The dosage was 5, 10, and 20 mg/Kg/day for each of compound () and tricholabdal B, respectively, and the average survival days at each dosage were determined.
These were compared with the average survival days in the control group to which only physiological saline was administered, and the rate of increase in survival (%) was calculated according to the formula below. Increase in survival rate (%) = average survival days of sample administration group - average survival days of control group / average survival days of control group x 100 The results are shown in the table below. 【table】

Claims (1)

[Claims] 1. General formula (In the formula, R ¹ means a methyl group or methylene group, R ² means hydrogen or a hydroxyl group, and R ³ means hydrogen or an acetoxy group.) 2 formulas The diterpenoid according to claim 1, which is represented by: 3 formulas The diterpenoid according to claim 1, which is represented by: 4 formula The diterpenoid according to claim 1, which is represented by: