JPH045229A - Carcinogenic preventive agent - Google Patents

Abstract

NEW MATERIAL:A compound expressed by the formula (R is 1-5C alkyl; n is 4-22). EXAMPLE:14-Methyl-1-pentadecanol. USE:A carcinogenic preventive agent effective against not only ascitic cancer and leukemia but also solid cancer. PREPARATION:For example, a natural substance such as wool grease is subjected to saponifying reaction in the presence of an alkali. An alcohol and a fatty acid are separated from the resultant substance after completing the reaction to provide wool alcohol, which is then subjected to molecular distillation to afford a low-boiling fraction. Fractionation is further carried out by reverse phase column chromatography and the fraction having the third highest polarity is subsequently fractionated by normal phase column chromatography and finally fractionated by high-performance liquid chromatography, etc., to afford the compound expressed by the formula.

Description

[Detailed description of the invention] [Industrial application fields] The present invention relates to a novel anti-carcinogenic agent.

[Prior art] The prevailing theory is that carcinogenesis occurs in two stages, involving an initiator and a promoter, but the mechanism of carcinogenesis in terms of its effects on the human body in daily life is unknown, and its prevention is still subject to empirical trials. The development of anti-cancer drugs has been significantly delayed compared to anti-cancer drugs, due to nothing but mistakes.

[Problems to be Solved by the Invention] First of all, anti-cancer agents must be more safe than anti-cancer and must have no side effects. Secondly, there is a need for the drug to be effective orally so that it can be easily administered in daily life.

Third, it is desired that the method not only be effective against specific experimental cancers, but also show a wide spectrum of effects on various types of cancers that are actually susceptible.

Based on these points, we believe that it is appropriate to develop anti-carcinogenic agents from among the "biological substances" that exist physiologically in the bodies of higher animals, and as a result of searching for sebum-derived substances, we found that
Among these, we found a substance that satisfies these conditions. These substances are thought to be useful not only for general carcinogenesis prevention, but also for preventing recurrence after cancer treatment is completed, and for preventing occupational cancer or cases where the cancer is genetically inherited in the family.

The compound of the present invention is chemically stable and resistant to cooking, so it can be put to practical use as a food additive, functional food, or health food.

Furthermore, since the compound of the present invention is a substance originally present in sebum, it is expected that the general public will be able to accept it without any resistance.

[Means for Solving the Problems] As a result of intensive research into anti-carcinogenic agents that have no side effects, the present inventors found that the general formula is CHi 0H (CH2) n CH2OH, where R has 1 to 5 carbon atoms. It has been discovered that an alkyl group and an isomonolol in which n is an integer of 4 to 22 have a remarkable anti-carcinogenic effect. That is, the present invention relates to the general formula (T); ■ CHI CH(CH2)11 CH2OH (1) (wherein, R is an alkyl group having 1 to 5 carbon atoms, and n is 4 to 22
The object of the present invention is to provide an anti-carcinogenic agent containing as an active ingredient an isomonolol represented by the integer .

[Example] In the present invention, the chain length of the alkyl chain of the isomonol is determined by the general formula (11, where n is an integer of 4 to 22, and R is an alkyl group having 1 to 5 carbon atoms. When n is less than 4, no anti-carcinogenic effect is exhibited at all.

Further, when the value is larger than 22, there is a problem that the anti-carcinogenic effect is not exhibited. When R is an alkyl group having 6 or more carbon atoms, there is a problem in that it does not exhibit anti-carcinogenic effects. In the general formula (I), the cancer prevention effect increases rapidly when the value of n is to-te, and when R is a methyl group or an ethyl group. Among them, the formula (■): CH3 C)lx CH(CH2) 12 CH2OH(II
) and the formula [11].

14-Methyl-1-hexadecanol represented by the formula %[1 shows particularly high anti-carcinogenic effects and is particularly preferred.

The isomonol used in the present invention can be obtained by decomposing natural products such as wax. The natural products used include waxes such as wool wax, spermaceti wax, beeswax, fresh white wax, and carnauba wax.

The isomonol used in the present invention can be obtained, for example, by the following method.

(Saponification reaction) For example, a natural product such as wool grease is suspended in water in the presence of a 1.18-01 alkali (NaOH), and a saponification reaction is carried out at 135±5°C with stirring under pressure for 3 hours. (using an autoclave).

(Separation of alcohol and fatty acids) H2O and CHI C0C2H5 (hereinafter referred to as M
EK) is added, transferred to a separatory funnel and heated to 70-75°C to extract and separate the alcohol into MEK. The solvent of the MEK solution of wool alcohol is removed under reduced pressure to obtain wool alcohol as a solid.

(Molecular distillation of wool alcohol) Solid wool alcohol is molecularly distilled to produce a low boiling point fraction (temperature:
<80°C, pressure crab IX to -2 Torr). This fraction is designated as MDl#e.

(Fractionation of MDI-Aic by reverse phase column chromatography) MDI-#c is fractionated into 6 fractions by reverse phase column chromatography (oven column).

Fractionation conditions packing material: ODS crushed, pore size 60, particle size GO/
200 meshes (product name: YMC-GEL, manufactured by Yamamura Chemical Research Institute) were removed at night: C11 (Js/CH).
JOH/H20-5/15/1 (volume ratio) The third eluted fraction from the beginning, in other words, the third most polar fraction is designated ODS$3.

(Fractionation of 0DStI3 by normal phase column chromatography) ODS$3 is fractionated into three fractions by normal phase column chromatography (oven column).

Fractionation conditions Filler Nizirika gel, pore size 60 people, TYPE 60 people 5PECIAL.

Particle size 100/200 mesh (product name: 5ILICA
R■, manufactured by Maillinckrodt, imported and sold by Dai-dai-
Chemical ■) Eluent: Liquid CHCl s / CHsC0CHz
-98/4 (volume ratio) Second solution CH30H Elute up to the second fraction with the second solution, and elute the third fraction with the second solution. The second eluted fraction was ODS$l3-
Set it to 2.

(Fractionation of ODS $3-2 by HPLC: Separation and preparative collection of isomonol) ODSt13-2 was subjected to high performance liquid chromatography (HPLC)
LC) to isolate the target compound.

11PLc condition column: TSK gel 0DS-120T (product name, Tosoh ■, (21.5 (inner diameter) x 300m
m)) Mobile phase: CI+30 H/H20-9515 (
Capacity ratio) Flow rate: 5. Oml/min Column temperature: room temperature Isomonol may be separated and fractionated by other methods. Other methods include, for example, Satoshi Taka, Makoto Yamanaka, Kikuhiko Saiki Moto, and Fumio Cyto-Allergens of Lanolin: Part I and■,
Part I: Isolation and Identification of the Allergens of Hydrogenated ψ Lanolin, Journal of the
Society of Cosmetic Φ Chemistry, 34
Volume, pages 99-11G (1983) (SATO9HI
TAKANO, MAKOTOYAMANAKA.

KIKUHIKOOKAMOTO, and FUMIO
5AITO, AIIergensorlanolin
: parts I and Il, PARTI
:l5OLATION AND IDENTIF
ICATION OF THEALLERGENS
OF HYDROGENATED LAN0L
IN, JOLIRNALOF THE 5OCIETY
OF CO8METICCHEMISTS, 34°p
99-116.1983).

Isomonols include, for example, when R is a methyl group, iso-〇M-OH[12-methyl-1-tridecanol]iso-〇ps-0)1 [13-methyl-1-tetradecanol], Iso-CI6-OH [14-methyl-1-pentadecanol], Iso-CI7-OH [15
-methyl-1-hexadecanol], iso-C+a -
Examples include OH[1[i-methyl-1-heptadecanol], and when R is an ethyl group, iso-〇+s -On [12-methyl-1-tetradecanol], iso-CI6 -OH[13-methyl-1-
pentadecanol], iso-CI7-OH[14-methyl-1-hexadecanol], iso-Cps -OH[
15-methyl-1-heptadecanol], iso-〇ps
-OH[1B-methyl-1-octadecanol] and the like.

These isomonols may be used alone or in combination of two or more.

To make the anti-carcinogenic agent of the present invention into a preparation for injection or infusion, Pluronic F-68 (trade name, manufactured by Asahi Denka Kogyo Kan Co., Ltd.), H
Add a surfactant such as CO-GO (trade name, manufactured by Nikko Chemicals) and disperse it using ultrasound, or make it into a liposome or oil-in-water emulsion, and add a preservative such as methyl p-hydroxybenzoate, lecithin, or linoleic acid. Stabilizers such as, non-aqueous vehicles such as coconut oil, suspending agents such as glucose may also be included.

In addition, for oral preparations, capsules suitable for intestinal absorption must contain a binder such as gelatin, a stabilizer such as magnesium stearate, an excipient such as lactose, and a disintegrant such as potato starch. Enteric coatings can be formed using cellulose acetate phthalate, methyl acrylate/methacrylic acid copolymers, etc. In addition, it can be formulated as granules, sustained-release implantable capsules, halved capsules, and nebulizer capsules.

The anti-carcinogenic agent of the present invention is administered parenterally, such as by intravenous, subcutaneous injection, or infusion, at a dosage of the active ingredient (adult body weight 1
kg, per day) 5-1200mg, especially 20-1200mg
The amount is preferably 3001 g, and for oral preparations such as capsules, the amount is preferably o, i to 40 g, particularly 0.5 to 8 g.

The anti-carcinogenic agent of the present invention is effective against not only ascites cancer and leukemia but also solid cancers, and has a wide range of indications including adenomas, squamous cell carcinomas, undifferentiated carcinomas, and sarcomas of various tissues. . In addition, it is effective not only on cancer-transplanted animals but also on cultured malignant cells of humans, mice, rats, hamsters, etc., so it has a direct cancer cell killing effect and is not species-specific.
It can be used in medicine and as a cancer chemotherapeutic agent for livestock and animals.

Furthermore, therapeutic effects have been observed not only by direct administration to the site of tumor implantation but also by remote administration. Toxic LDs.

is 5.1~Lag/kg by subcutaneous injection in rats, and 1~Lag/kg.
No side effects were observed even after continuous administration of 2 g/kg for 10 days.

The method for producing isomonol, which is the active ingredient of the anti-carcinogenic agent of the present invention, will be explained below with reference to Examples, but the present invention is not limited to these Examples.

Example 1 200 g of wool alcohol obtained by saponifying wool grease was subjected to molecular distillation to obtain a low boiling point fraction HDl-M.
IC (temperature: <80℃, pressure crab lXl0-2 Torr
) gained 15.2g.

MDI-Mc was subjected to reverse phase column chromatography (oven column), and the mobile phase was CHCl3/CHxOH/H.
It was divided into 6 fractions at a ratio of 20-5/15/l (volume ratio). Of these, the third eluted fraction (0DS113) was 1.68
I got it.

This ODS$3 was subjected to normal phase column chromatography (oven column), and mobile phase liquid: CHCl s
/C) 13COCH3-96/4 (volume ratio), second liquid: 3 fractions with Cll30H.

Of these, 0.96 g of the second eluted fraction (ODS $3-2) was obtained.

OD! 143-2 by high performance liquid chromatography (HP
The target compound was isolated by fractionation using LC.

14-methyl-1-pentadecanol, 14-methyl-
1-hexadecanol, 16-methyl-1-heptadecanol, 16-methyl-1-octadecanol, 18-
Methyl-1-nonadecanol was obtained.

It was then confirmed by capillary gas chromatography that each isomonol was clearly isolated.

+3C-NMR and GC-MS were used to determine the structure, and the isolated substance was identified to be an isomonol group represented by the general formula of the present invention.

For 14-methyl-1-pentadecanol, the C-NMR spectrum (100,40 MHz) is shown in Figure 2.

CD(J3)6 (ppIll) is shown, and C-MS data are shown in FIG. 3 and FIG. 4. For 14-methyl-■-hexadecanol, see Figure 5.
C-NMR spectrum (100,40Mt (zSCDC
I3) δ (ppm) is shown in Figures 6 and 7.
C-MS data is shown.

In addition, the data in FIGS. 3 and 6 are based on the mode or El,
The data in Figures 4 and 7 were measured under the conditions that the ionization voltage was 70 eL and the ion source temperature was 250°C. It was measured under the condition of ℃.

The isomonol isolated here was subjected to an in vivo test as a carcinogen.

Test Example 1 Using 14-methyl-1-pentadecanol as a carcinogenic agent, 6-week-old SD female rats (90 rats) were treated as follows.

Non-administration group = 30 animals Oriental yeast as basic feed■
They were fed MP powdered feed manufactured by

Administration group A: 30 animals As a test feed, a mixture of the above carcinogenic agent mixed with Pluronic F-68 at a rate of 50 μg/kg after emulsification was fed to the basic feed continuously (for 7 days).

Administration group B: 30 animals As test feed, a mixture of the above-mentioned anti-carcinogenic agent with Pluronic F-68 at a rate of 250a+g/kg after emulsification was continuously fed (for 7 days) to the basic feed.

There was no significant difference in the amount of food intake between the administration groups.

After the treatment, 5 μ/mouse of benzanthracene (DMBA) was administered.
: a carcinogen) was administered by subcutaneous injection into the mammary gland at 0.1 ml/mouse of a 50 II g/ml solution of benzanthracene, and then the respective administration groups were fed the respective diets for 20 weeks.

There was no significant difference in the amount of food intake between the administration groups.

After completion of the above treatment, the test rats were sacrificed and the development of breast cancer was observed. The results are shown in Table 1.

Table 1 Test Example 2 Male Wistar rat (8-9 weeks old, body weight flog) 4
187 μg/ml N-methyl-
Evening drinking water and feed containing N-nitro-N-nitrosoguanidine (MNNG: carcinogen) (dried pellets CE-2,
(manufactured by CLEA Japan) was given for 210 days.

On day 210, adenoma-like hyperbrassia was observed in the stomach.

After the 211th day, the rats were divided into 5 groups and subjected to the following treatments.

Non-administration group: 28 animals Administration group A: 28 animals Compound A of the present invention (see below, the same applies hereinafter) 250 mg/kg feed x 126 days continuous oral administration Administration group: 8.8 animals Compound of the present invention 8 250 mg/kg
Feed x 126 days continuous oral administration Group C: 8 animals Compound of the present invention C 250 mg/kg Feed x 126 days continuous oral administration Group D = 8 animals Compound D of the present invention 250 mg/kg
Feed x 126 days continuous oral administration A: 14-methyl-1-pentadecanol B: 14-methyl-1-hexadecanol C: 16-methyl-1-heptadecanol D: 16-methyl-1-octa Rats that had survived for 337 days with decanol were killed by stratification, and the stomach was (1) visually observed using Borra+Ann.
Classification and (a) Histological observation (hematoxylin and eosin staining or ASAN staining) were performed as follows, and the results are shown in Table 2.

Grade 0: Gastric carcinoma not observed Grade I: Gastric carcinoma, intramucosal grade ■: Gastric carcinoma, infiltrating into the submucosal layer ■: Gastric carcinoma, invading into the muscle layer, recommended membrane grade ■: Gastric carcinoma invading even in the vicinity Test Example 3 Five groups of 6-week-old F344 male rats were given 0.05% (wt%, same hereinafter) N-butyl-N-( 4
-Hydroxybutyl) nitrosamine (BBN: bladder irritant) aqueous solution was administered in a water bottle for 4 weeks, then 3
For two weeks, 5% sodium erythorbate aqueous solution as a promoter (carcinogenic substance) and compounds of the present invention A-D (see below) were administered at 250a+g/k to an MP powdered feed manufactured by Oriental Yeast ■ as a basic feed.
A feed containing 1.5 g was administered.

A 214-methyl-1-heptadecanol B: 1
4-Methyl-■-hexadecanol C: 16-methyl-
There was no significant difference in the amount of 1-heptadecanol D = 18-methyl-1-nonadecanol feed between the administration groups.

After the above treatment, the test rats were sacrificed and the development of bladder mucosal lesions was observed.

The results are shown in Table 3.

[Effects of the Invention] The anti-carcinogenic agent of the present invention exhibits an excellent anti-carcinogenic effect, and its active ingredients are derived from living organisms (animals such as K),
It does not show any serious side effects to living organisms.

[Brief explanation of the drawing]

FIG. 1 is a flowchart showing the method for producing isomonol according to the present invention. Figure 2 shows '3C of 14-methyl-I-pentadecanol.
-NMR spectrum. Figures 3 and 4 are mass spectra of 14-methyl-1-pentadecanol. Figure 5 shows 13C of 14-methyl-1-hexadecanol.
-NMR spectrum. Figures 6 and 7 are mass spectra of 14-methyl-1-hexadecanol.

Claims (1)

[Claims] 1 General formula (I): ▲There are numerical formulas, chemical formulas, tables, etc.▼(I) (In the formula, R is an alkyl group having 1 to 5 carbon atoms, and n is 4
An anti-carcinogenic agent containing as an active ingredient an isomonolol (representing an integer from 22 to 22). 2. In the general formula (I), R is a methyl group or an ethyl group, and n is an integer of 10 to 16. Claim 1
The listed cancer prevention agents. 3. The anti-carcinogenic agent according to claim 1, wherein the active ingredient is 14-methyl-1-pentadecanol represented by formula (II): ▲Mathematical formula, chemical formula, table, etc.▼(II) 4. The anti-carcinogenic agent according to claim 1, wherein the active ingredient is 14-methyl-1-hexadecanol represented by formula (III): ▲Mathical formula, chemical formula, table, etc.▼(III).