JPS5832874A - Diterpenoid and antitumor agent comprising it as active ingredient - Google Patents

Abstract

NEW MATERIAL:A diterpenoid shown by the formulaI(R<1> and R<2> are H or hydroxy; R<3> is H or acetoxy). EXAMPLE:Trichorabdal A shown by the formula II. USE:An antitumor agent. PROCESS:Leaves of Isodon Kameba Okuyama are dried, extracted with an organic solvent such as an ether, acetic acid ethyl ester, acetonitrile, acetone, methanol, ethanol, dichloromethane, chloroform, etc. under reflux, and concentrated to give a concentrated residue, which is washed with water, saccharides are removed, and the resultant substance is subjected to column gel chromatography, to give a diterpenoid shown by the formulaI.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel diterpenoid and an antitumor agent containing the same as an active ingredient.

The present inventors have been conducting repeated research on the extract from Kurobananahikiokoshi, a plant of the genus Yamahathuka of the family Sinaceae, located in Kiyamahiko, Hyogo Prefecture, and have succeeded in isolating novel diterbenoids.Moreover, these diterpenoids have excellent anticancer effects. The present invention was completed based on this discovery.

The novel diterpenoids of the present invention are represented by the following general formula.

(In the formula, R1 and R2 are hydrogen or hydroxy groups, R3 is water 1
A large number of diterbenoids have been isolated from plants of the Lamiaceae family and genus Yamahata, and compounds with physiological activity are also known. The Black Banana Hikiokoshi used in the present invention belongs to the Lamiaceae family, and is a perennial plant that grows naturally in the mountains and is also known as Enmeiso. Oridonin, a diterpenoid with anticancer effects,
It is known that lasiocaulin, enmain, etc. exist. The compound of the present invention is a novel compound different from these diterpenoids, and has a superior antitumor effect than these conventionally known diterpenoids.

The diterpenoid of the present invention is specifically represented by the following structural formula.

5- Tricholabdal A Tricholabdal B Tricholabdal C Tricholabdal D To produce the nositerpenoids of the present invention, the leaves of the black-and-white plant are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried, and the leaves are dried. , extracted under reflux with an organic solvent such as methane dichloride or chloroform, and then concentrated. After washing the residue with water to remove sugars,
Obtained by column chromatography.

The details of the above production method and the physical properties of the compound of the present invention obtained thereby will be clarified in the Examples below.

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, it is prepared in the form of tablets, capsules, granules, powders, liquids, etc., and for parenteral administration, it is prepared in the form of injections, tablets, 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, 11' starch, talc, magnesium stearate, crystalline cellulose, methyl cellulose, carboxymethyl cellulose, glycerin, sodium alginate, gum arabic, etc., and binders include polyvinyl alcohol, holvinyl ether, ethyl cellulose, Gum arabic, shellac, sucrose, etc. can be used, lubricants include magnesium stearate, talc, etc., and colorants and disintegrants that are commonly known can be used. Additionally, 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, a pH adjuster, a buffer, a stabilizer, a tonicity agent, a local anesthetic, etc. are added to the compound of the present invention, and an injection for subcutaneous, intramuscular, or intravenous use is produced by a conventional method. can do.

As a base for manufacturing the planing, an oily base such as cacao butter, polyethylene glycol, lanolin, fatty acid triglyceride, Witepsol (registered trademark: Dynamite Nobel), etc. can be used.

The dosage of the preparation thus prepared depends on the patient's symptoms, body weight,
It varies depending on the age etc. - Although it cannot be generally limited, the compound of the present invention is usually administered in doses of about 50 to 1,000 doses per day for adults.
The amount should preferably be in the range of mf/, which is usually 1
It is preferable to administer the drug in 1 to 4 divided doses per day.

Next, the production method, physical properties, and pharmacological effects of diterpenoids, which are the compounds of the present invention, will be explained in more detail with reference to Examples.

Example Kyoko Prefecture Ice Birth Dry leaves of Black Banana Hikiokoshi 9- 1.52Y4! is refluxed with 12/ml of methanol for 12.5 hours. The reflux liquid was concentrated to obtain a residue 1289. This residue was mixed with methanol 8I! After dissolving the impurities in hexane II
! Wash 8 times. After concentrating the methanol layer, it was dissolved in 81 ml of ethyl acetate and washed 8 times with 800 ml of water each time to remove water-soluble components. Concentrate the ethyl acetate layer to obtain a residue 505'
I got it. This residue was subjected to the following first column chromatography for crude separation.

First column chromatography 50 g of the residue obtained above was dissolved in a mixture of methylene chloride and acetone (7:8 volume ratio), and silica gel (70-28
0 mesh) 1.81$ column (diameter 10cm x length 4
5 (m), and the developing solvent was sequentially changed to obtain the following two components.

10- Elution fraction Developing solvent Target product 2500-1
4500ml! Methylene chloride -1
4500-18500ml! methylene chloride
Trico Labdal A2800 G-29500ml
〃-2950ω-86500n17? Both acetone and trichorabdal D and above were further separated again as follows.

Trichorhabdal A: 8.94y of trichorhabdal A obtained in the first column chromatography was added to silica gel 60.
Column chromatography of F, eluted with methylene chloride, 600 ml to 750 ml of eluate! The fractions were concentrated to obtain crude crystals, which were recrystallized from methanol to obtain 48 mg of trichotorabdal A in the form of needle-like crystals.

Melting point: 198-201°C 1S [α) Knee 68.9° (cmO, 01, ethanol) Ultraviolet absorption spectrum λmax (ethanol) 282nm (#8
500) Infrared absorption spectrum v m5zx (KBr) 8600,8450
．． 2820.2750°1?40, 1710, 16
40, 1275an nuclear magnetic resonance spectrum (ds-pyridine, 25°C) 629m0.97.1.00 (ea
ch 81 (, s, -CH8) 2.90 (IH, d,
J-4Hz, 5-H) 8.10 (IIl,, dd,
J=10.4Hz, 1B-H)8.50 (IH,d
, J=12Hz, 14-Ha)4.62 (2H, m
, 2O-HAand 1l-H)5.14 (IH
, d, J=12Hz, 20-H/)5, f35.5.9
9 (esich IH, s, 17-It) 6.82
(IH, d, J-4, -0H) 10.04 (IH, d,
J=4Hz, -CIO) Tricholabdal B Residue 16.775 obtained by concentrating both fractions containing Tricholabdal B obtained in the first column chromatography under reduced pressure
'Silica gel column chromatography (silica gel 8
0(1) and developed with a mixture of methylene chloride and acetone (9:l volume ratio). Collect 150 ml for each portion, 6
Concentrate both fractions and recrystallize from methanol to obtain columnar or prismatic tricholabdar B20.
5m5+ was obtained.

Melting point: 161-162℃ 18-! 6 [σ): -tgo, so(cmO, 86
, ethanol) UV absorption spectrum λmax (ethanol) 282 nm (g=9
100) Infrared absorption spectrum v max (KBr) 8600.8450.28
20.2720°1740.1710,1640.12
20an nuclear magnetic resonance spectrum (d+s-pyridine, 6
0℃) 899m1.19 (8H, s, -CHa) 2.00 (8B, s, -0COCHs) 8
．． 11 (IH, d, J=4Hz, 5-H)8.11
(IH, m, 18-H) 8.50 (IH, d, J=12Hz, 14-Ha)
4.08, 4.16 (each IH, ABQ, J-
12H2,19-Hs) 4.50 (IH, m, 1l
-H) 4.50, 5.15 (each IH, ABqJ=1
2Hz, 20-Hri 14-5.4G, 6.05 (eacb IH,s
, 17-Hg) 6.40 (LII, br, s,
-0f()10.19(IH, d, J=4Hz, -CH
o) Tricholabdal C 8.4v of the residue obtained by concentrating both the 9th to 14th fractions obtained by silica gel column chromatography of the above Tricholabdal B was subjected to silica gel column chromatography ((Silica gel 2009)) and diluted with methylene chloride. -Acetone mixture (
9:1 volume ratio) and expand to 250 ml! Aliquots were taken. @26th to 84th fractions were concentrated to obtain amorphous tricholabdar C1,72.

Ultraviolet absorption spectrum Infrared absorption spectrum pmax (KBr) 8450, 2750, 1745
．． 1715°1640.1280cm Nuclear magnetic resonance spectrum (CDC/, , 25℃) 89
9m1.21 (8H,s, -CHg) 2.06 (8H,s, -0COCHs)ca
2.80 (IH, m, -0H) 2.86 (I
H, d, J = 8.4Hz, 5-H) 8.10 (IH
, dd, J=9and8T(z,18-H)8.65
(IH, m, 1l-H) 4.07, 4.20 (each IH, ABq,
J=12Hz, 2O-Hg) 4.58, 4.69
(each IH, ABq, J=12Hz, 19
-Hg) 5.50, 6.01 (each IH,s
, 17-Hg) 9.85 (IH, d, J=8
．． 4Hz, -CHO) Tricholabdar D No. 65 to 72 obtained in the first column chromatography
Both fractions were concentrated to obtain crude crystals, which were recrystallized from methanol to form needle-like crystals of Tricolabdal D50mf!
I got it.

Melting point: 218-216℃ 15 [α: l knee 89.2° (c=gO, o07.Ethanol) Ultraviolet absorption spectrum λmaz (ethanol) 280nm (g-187
00) Infrared absorption spectrum v max (KBr) 8550.8470,28
40,050°1745.1780.171G, 164
0°240cm nuclear magnetic resonance spectrum (drh-pyridine, 60'O)
899m1.40 (8H, s, -CHg) 17- 2.02 (8B, s, -0COCHI) 8.12
(1B, dd, J=10and4Hz, 1B-H)
8.55 (IH, d, J-4Hz, 5-H) 8.5
4 (IH, d, J=12Hz, 14-Hcr)4.
20 (IH, m, 8-H) 4.82, 4.66 (each IH, ABq,
J=11.81tg.

19-Hl or 2O-Hl) 4.58 (IH, m, 1l-H) 4.8 B, 15.16 (each IH, AB
q, J-11,7Hz20-Hi or 19
H! ) 5.42, 6.05 (each IH,
17-Hg)6.54 (2H,br,s,-0H
) 10.22 (IH, d, J=4Hz, -CHo)
Comparative studies were carried out with the known compound oridonin, which is a diterbenoide of the same genus as the tricholabdal B and tricholabdal 18-ru B of the present invention and has already been reported to have anticancer activity.

2×10′ Ehrlitsu rutinoma vesicles/mouse were implanted intraperitoneally into male ddY mice (25-28y). Tricolapdal B or oridonin was dissolved or suspended in physiological saline and administered at a dose of 0.1 ml per group of 7 mice.
The drug was administered intraperitoneally once a day for 7 consecutive days starting the day after tumor implantation at a volume ratio corresponding to the body weight of IOy mice. The dosage was 10.15 for each of tricholabdal B and oridonin.
and 20 m9A! /day, the average number of days of survival for each dose was determined, and these were compared with the average number of days of survival in a control group to which only physiological saline was administered, and the rate of increase in survival (ri) was calculated according to the following formula.

Table 1 (Continued) Kazuo Wakasugi, Commissioner of the Japan Patent Office1, Indication of the case, 1982 Patent Application No. 129840 20 Title of the invention: Diterpenoids and antitumor agents containing the same as active ingredients3.
Relationship with the case of the person making the amendment Patent applicant Taiho Pharmaceutical Co., Ltd. 4, Agent Address: 1-2-8 Maruville, Yotsutanezaki, Kita-ku, Osaka 530 Tel: 06
(365) No. 0170 (Main) Voluntary Issue 6. Number of Inventions Increased by Amendment Contents of Amendment 1 The scope of claims is revised in the attached document.

2 Specification, page 6, line 10 [ 3 Specification, page 6, lines 3-4 "Trichorapugur C"" 4 The following is added next to Nuclear Magnetic Resonance Subek 1 Hell Dark on page 16 of the specification.

"Amorphous tricolapgol C could be recrystallized from ethyl acetate. Melting point 1/13.5~+45°C" (
2) 2-(3) A diterpenoid represented by the general formula (1) ('ill'n', R2 means hydrogen or a hydroxy group, and 3 means hydrogen or an acetoxy group).

The diderpenoid according to claim 1, which is represented by formula (2).

1- The diterpemade according to claim 1, which is represented by formula (5).

The diterpenoid according to claim 1, which is represented by formula (4).

The diterpenoid according to claim 1, which is represented by:

(6) An antitumor agent containing as an active ingredient a diterpenoid represented by the general formula (wherein R2 means hydrogen or a hydroquine group, and R3 means hydrogen or an acetoxy group).

Claims (1)

[Scope of Claims] (1) A diterpenoid represented by the general formula [wherein R1 and R2 are hydrogen or a hydroxy group, and R8 is hydrogen or an acetoxy group]. The diterpenoid according to claim 1, which is represented by formula (2). The diterpenoid according to claim 1, which is represented by: (The diterpenoid according to claim 1, which is represented by the formula 41. The diterpenoid according to claim 1, which is represented by the formula (5). (6) General formula (wherein R", T?" are Hydrogen or hydroxy group, R8
An antitumor agent containing as an active ingredient a diterpenoid represented by halogen or acetoxy group.