JPH045228A - Carcinostatic agent and carcinostatic enhancer - 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 carcinostatic agent and carcinogenic enhancer without exhibiting serious side effects on living bodies, because the compound is derived from the living bodies. PREPARATION:For example, a natural substance such as wool grease is subjected to saponifying reaction in the presence of an alkali. After completing the reaction, a solvent is removed to afford wool alcohol as a solid substance. The resultant wool alcohol is then fractionated by molecular distillation and column chromatography and further fractionated by high-performance liquid chromatography, etc., to afford the objective compound.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to novel anticancer agents and anticancer enhancers.

[Conventional technology] Conventionally, anticancer drugs include alkylating agents that exert anticancer properties by alkylating biopolymers with important functions such as nucleic acids and enzymes, and alkylating agents that inhibit nucleic acid metabolism. antimetabolites, which are involved in the biosynthesis of nucleic acids in cells before cell division;
There are anticancer antibiotics that have a cell-killing effect on actively dividing and proliferating cells, as well as plant-derived anticancer substances and hormones. On the other hand, amphotericin B is being researched as an anticancer enhancer that is effective in improving the absorption of drugs that are difficult to absorb by cancer cells and strengthens the anticancer effects of anticancer drugs such as pleomycin and adriamycin. has not reached significant effect.

[Problems to be Solved by the Invention] Conventional anticancer drugs all have side effects and few have satisfactory efficacy, and there is a demand for the development of better anticancer drugs. In addition, in order to fully exhibit the effects of anticancer drugs, excellent anticancer enhancers are required.

[Means for Solving the Problems] As a result of extensive research into drugs without side effects, the present inventors found that the general formula is 0H3CH(CH2)I, CH2OH, where R is 1 carbon number. It has been discovered that isomonols having an alkyl group of ˜5 and n being an integer of 4 to 22 have a remarkable anticancer effect. That is, the present invention provides compounds of the general formula (I): (in the formula , R represents an alkyl group having 1 to 5 carbon atoms, and n represents an integer of 4 to 22. The present invention also provides an anticancer enhancer containing the isomonol as an active ingredient.

[Example] In the present invention, the chain length of the alkyl chain of the isomonol is such that n in the general formula (1) 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 anticancer effect or anticancer enhancing effect is exhibited.

Moreover, when it is larger than 22, there is a problem that the anticancer effect and the anticancer enhancing effect are not exhibited.

When R is an alkyl group having 6 or more carbon atoms, there is a problem that it does not exhibit anticancer effect or anticancer enhancing effect. In the general formula (Il), the anticancer effect and the anticancer enhancing effect are rapidly increased when the n(7) value is 10 to 16, and for the same reason, R is a methyl group or an ethyl group.

Among them, 14-methyl-1-pentadecanol of the formula (■): CH3CHxCH(CH2)r2CH20H(I) and the formula (I[l).

CH3 CH3CH2CH(CH2) 12 CH2OH(I[
14-Methyl-1-hexadecanol represented by [l shows particularly high anticancer effect and anticancer enhancing effect, and is preferable for ().

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 1.18 times mol of alkali (NaOH), and a saponification reaction is carried out under pressure at 135±5°C with stirring for 3 hours. (using an autoclave).

(Separation of alcohol and fatty acid) Add 820 and C)IsC0C2)Is (hereinafter referred to as MEK) to the saponification reaction product (mixture of Na salt of higher fatty acid and higher alcohol), transfer to a separatory funnel, and add 70~7
The alcohol is extracted and separated into MEK by heating to 5°C.

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 I X 10-2 Torr) is obtained. This fraction is designated as MDI-M'c.

(Fractionation by reverse phase column chromatography of MDI-Ai)c) Fractionate into 6 fractions by reverse phase column chromatography (oven column) of MDI-M)e.

Fractionation conditions packing material: ODS crushing wire pore size 60, particle size 60/
200 mesh (product name: YMC-GEL, manufactured by Yamamura Chemical Research Institute) Eluent acid diCHCR3/CH30H/H
20=5/15/1 (volume ratio) The third eluted fraction from the beginning, in other words, the third most polar fraction is designated as ODS 3.

(Fractionation of 0DStt3 by normal phase column chromatography) ODSS3 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 Mal I Incrodt, imported and sold by:
Daiichi Chemical #) Eluent: Liquid CH (J s / CH3COCH3
-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. Both the second eluted fractions were 0DSJt3-
Set it to 2.

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

HPLC condition column: TSK get 0DS-120T (product name, manufactured by Tosoh ■, (21.5 (inner diameter) x 300 mm
)) Mobile phase: CHsOH/H20-9515 (volume 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 Takahamakoto ◆ Yamanaka, Kikuhiko Naka Okamoto and Fumio Cyto-Allergens of Lanolin: Bart I and ■,
Part I: Isolation and Identification of Allergens of Hydrogenated Lanolin, Journal of the
Society of Cosmetic Chemistry, 34
Volume, pp. 99-11B (1983) (SATO3HI
TAKANO, MAKOTOYAMANAKA.

KIKUHIKOOKAMOTO, and FU former O5
AITO, Allergensof' 1anolin
: parts I and II, PART
I :l5OLATION AND IDENTIFI
CATION OF THEALLERGENS OF
HYDROGENATED LAN0LIN, JOU
RNALOF THE 5OCIETY OF CO3
METICCHEMISTS, 34゜p99-116
, 1983).

Isomonols include, for example, when R is a methyl group, iso-CI4-OH[12-methyl-1-)ridecanol], iso-〇+s-OH[13-methyl-1-tetradecanol], iso-CI4-OH[12-methyl-1-tetradecanol], -CI6-OH[14-methyl-
1-pentadecanol] iso-CI7-OH [15-methyl-1-hexadecanol coiso〇+s -OH[
1B-methyl-1-heptadecanol], and when R is an ethyl group, iso-CI5-Of([12-methyl-1-tetradecanol], iso-〇ps -OH[ 13-methyl-1-
pentadecanol]iso-CI7-OH[14-methyl-1-hexadecanol coiso-〇ps-OH[15
-Methyl-1-hexadecanolcoiso-CI9-OH
Examples include [1B-methyl-1-octadecanol].

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

To prepare the anticancer agent and anticancer enhancer of the present invention for injection or infusion, use Pluronic F-68 (trade name, manufactured by Asahi Denka Kogyo ■), HCO-130 (trade name, manufactured by Nikko Chemicals Co., Ltd.). Add a surfactant such as, disperse using ultrasound, or form a liposome or oil-in-water emulsion, preservative such as methyl p-hydroxybenzoate, stabilizer such as lecithin, linoleic acid, or non-aqueous vehicle such as coconut oil. Suspending agents such as , glucose, etc. may 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, suppositories, nebulizers, and buccal preparations.

When the anticancer agent of the present invention is administered parenterally, such as by intravenous, subcutaneous injection, or infusion, the dosage of the active ingredient (1 kg of adult body weight)
, per day) 5-1200■, especially 20-300■
is preferable, and for oral administration agents such as capsules, 0.1 to
40g1 is particularly preferable, 0.5 to 8g. When used as an anticancer enhancer, the parenteral dosage is 0.45~
102■, preferably 1 to 28 mg s For oral administration, 0.005 to 5.2 g, preferably 0.07 to 1
．． It is 2g.

The anticancer agent and anticancer enhancer of the present invention can be used to treat ascites cancer,
It is effective against not only leukemia but also solid cancers, and has a wide range of indications including adenocarcinoma, squamous cell carcinoma, undifferentiated cancer, and sarcoma of various tissues.

In addition, it is effective not only against cancer-transplanted animals but also against 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 as a cancer chemotherapeutic agent in 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.

was subcutaneously injected into rats at a dose of 5.1 to 17 g/kg, and 1 to 17 g/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 anticancer agent and anticancer enhancer of the present invention, will be explained below by giving 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 MDI-A.
Rc (temperature: <80℃, pressure crab 1X1O-2Torr)
I gained 5.2g of L.

MDI-M'e was subjected to reverse phase column chromatography (oven column), and the mobile phase was CHCj3/CH30H/
It was fractionated into 6 fractions using H2O-5/15/1 (volume ratio). Of these, the third eluted fraction (ODS$3) was
I got it.

This ODS$3 was subjected to normal phase column chromatography (oven column), and mobile phase liquid: CHCl 3/
CH3COCH3-98/4 (volume ratio), second liquid:
It was fractionated into 3 fractions with CH30H.

Among these, the second eluted fraction (0DSt13-2)
0.98g was obtained.

0DStI3-2 was subjected to 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.

C-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.

FIG. 2 shows the 13C-NMR spectrum (100,40 MHz) of 14-methyl-1-pentadecanol.

CDCNx) δ (ppm), and GC-MS data are shown in FIGS. 3 and 4. Also 14-methyl-1-
Regarding hexadecanol, Fig. 5 shows the 13C-NMR spectrum (100,40MHz SCDCa3) δ (p
pm), and GC-MS data are shown in FIGS. 6 and 7.

Note that the data in FIGS. 3 and 6 are for modes El,
Measurements were made under the conditions of an ionization voltage of 70 eV and an ion source temperature of 250°C. The data in Figures 4 and 7 are based on CI mode, isobutane reaction gas, ionization voltage of 200 eV, and ion source temperature of 250 °C. It was measured under the condition of ℃.

The isomonol isolated here was tested as an anticancer agent and an anticancer enhancer.

Test Example 1 106 Ehrlich ascites cancer cells were inoculated into the peritoneal cavity of a 5-week-old ddY mouse, and 24 hours later, 50111 g of each sample was added to a 0.25% (wt%) Pluronic F68 physiological saline solution. The suspension at a concentration of ml was injected intraperitoneally to 10 animals per group for 5 days at lomg/)cg/day.

The control group administered the same wave without the sample had an average survival period of 14.0 days after inoculation, whereas 14-methyl-1-
Pentadecanol was administered for 60 days or more, 14-methyl-1-hexadecanol was administered for 57.5 days, and a mixture of these equal amounts was administered for 60 days or more, indicating a significant survival effect.

Test Example 2 106 adenocarcinoma 755 cells were subcutaneously transplanted into the back of a 6-week-old mouse, a one-generation hybrid of C57BL/8 and DBA/2, and 24 hours later, each cell was injected into a 0.25% HCO-30 saline solution. A suspension of the sample at a concentration of 50 μ/ml was subcutaneously injected to 8 animals per group at a dose of 10 mg/kg/day for 5 days. Ten days after transplantation of adenocarcinoma 755 cells, the mice were sacrificed and the tumors were excised.

The mean tumor weight (g) was 16- compared to 6.5 in the control group.
Methyl-1-heptadecanol and 16-methyl-1
- 1.6 and 2.0% for the octadecanol-treated groups, respectively.
3, and a significant tumor suppressive effect was observed.

Test Example 3 Using three different types of cancer cell lines (mouse ascites cancer Ehrlich cells, human lung carcinoma 549 cells, and mouse neuroblastoma NAs-1 cells), 14-methyl-1-pentadecanol and The growth inhibitory effect of 18-methyl-1-nonadecanol was investigated. As a result, the above 2
The different isomonols were effective against all three types of cancer cells.

When treated at IO μM for 6 hours, these isomonols reduced the colony formation rate of human lung carcinoma A349 cells by 2.
2X10-3 and 4. It was suppressed to lX10-3.

In addition, these isomonols inhibited the cell proliferation of Ehrlich cells by 4.5x10 after treatment at 10gM for 5 days.
-3 and 1.5X10-2.

The proliferation number of mouse neuroblastoma NAs-1 cells was 7.
It was suppressed to 8X10-3 and 6.5X10-3.

Test Examples 4 and 5 and Comparative Test Example 1 When bleomycin is used alone and when pleomycin and 14-methyl-1-pentadecanol or 14-methyl-1-
When hexadecanol was used in combination, the cell killing effect (IC9o) was investigated using mouse ascites cancer Ehrlich cells and human lung cancer type A349 cells. The results are shown in Table 1.

[Margins below] As is clear from the results shown in Table 1, when the isomonol of the present invention is used in combination with an anticancer drug, compared to when the anticancer drug is used alone. It can be seen that the cell killing effect is significantly improved.

Test Example 6 and Comparative Test Example 2 When pleomycin (60 μg/ml) and 14-methyl-1-pentadecanol (0.05 μM) were used together, the amount of pleomycin taken into Ehrlich cells was measured over time. I looked into it. The results are shown in Table 2.

[As is clear from the results shown in Table 2 below, when the isomonol of the present invention is used in combination with an anticancer drug, compared to when the anticancer drug is used alone, It can be seen that the amount of anticancer drug delivered into Ehrlich cells increases in a short period of time.

Test Example 7 and Comparative Test Examples 3 and 4C57BL/
106 Ehrlich ascites cancer cells were inoculated intraperitoneally into a 5-week-old male mouse, and after 24 hours, 0.2
50μ of pleomycin in 5% HCO-30 saline
solutions prepared in g / ml as well as 0,25%) IC
Pleomycin 50μg/O-6CI saline
ml and 14-methyl-1-pentadecanol at 5 μg/ml.
The latter was intraperitoneally injected at a rate of 11 kg/day for 5 days. Next, the number of days the injected mice survived was determined. The results are shown in Table 3.

Table 3 shows that the group administered with the pleomycin solution that does not contain the anticancer enhancer of the present invention (Comparative Test Example 3) had an average survival period of 33.3 days after inoculation, whereas It can be seen that in the group to which a pleomycin solution containing isomonol as an anticancer enhancer (Test Example 7) was administered, the average survival period after inoculation was 54.3 days.

Therefore, it is recognized that the isomonol of the present invention is useful as a control enhancer.

[Effects of the Invention] The anticancer agent and anticancer enhancer of the present invention exhibit excellent anticancer effects and anticancer enhancing effects, and furthermore, their active ingredients are derived from living organisms (higher animals). The drug does not show any serious side effects.

[Brief explanation of the drawing]

FIG. 1 is a flowchart showing the method for producing isomonol according to the present invention. Figure 2 shows "C" of 14-methyl-1-pentadecanol.
-NMI? It is a 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 anticancer drug 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
Anticancer drugs listed. 3. The anticancer agent according to claim 1, wherein the active ingredient is 14-methyl-1-pentadecanol represented by the formula (II): ▲Mathematical formula, chemical formula, table, etc.▼(II). 4. The anticancer drug according to claim 1, wherein the active ingredient is 14-methyl-1-hexadecanol represented by formula (III): ▲Mathematical formula, chemical formula, table, etc.▼(III). 5 General formula (I): ▲Mathematical 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 anticancer enhancer containing as an active ingredient an isomonolol represented by the formula (representing an integer from 22 to 22). 6. In the general formula (I), R is a methyl group or an ethyl group, and n is an integer of 10 to 16. 5.
The anticancer enhancer mentioned above. 7. The anticancer enhancer according to claim 5, wherein the active ingredient is 14-methyl-1-pentadecanol represented by the formula (II): ▲Mathematical formula, chemical formula, table, etc.▼(II). 8. The anticancer enhancer according to claim 5, wherein the active ingredient is 14-methyl-1-hexadecanol represented by the formula (III): ▲Mathematical formula, chemical formula, table, etc.▼(III).