JPH0772134B2 - Anti-cancer agent and anti-cancer enhancer - Google Patents

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel anti-cancer agent and anti-cancer enhancer.

[0002]

2. Description of the Related Art Conventionally, carcinostatic agents are alkylating agents that exert carcinostatic properties by alkylating biopolymers having important functions such as nucleic acids and enzymes, and antimetabolites that inhibit nucleic acid metabolism. Drug, a cell division toxin involved in the biosynthesis of cell nucleic acid, and an anticancer antibiotic that shows a cell-killing effect on cells that are proliferating and proliferating, as well as a plant-derived anticancer agent and hormone drug. There is. On the other hand, amphotericin B and the like have been studied as anti-tumor enhancers that are effective in improving absorption of drugs that are difficult to be absorbed by cancer cells and strengthen the anti-cancer effects of anti-cancer agents such as bleomycin and adriamycin. Not very effective.

[0003]

All of the conventional carcinostatic agents have side effects, and few of them have satisfactory efficacy, and there is a demand for the development of superior carcinostatic agents. In addition, an excellent anticancer drug is required to fully exert the effects of the anticancer drug.

[0004]

Means for Solving the Problems As a result of earnest research on an anticancer agent having no side effect, the present inventors have found that the general formula is

[0005]

[Chemical 11]

It was discovered that an alkyl group having 1 to 5 carbon atoms and a higher fatty acid in which n is an integer of 4 to 22 has a remarkable carcinostatic action. That is, the present invention has the general formula (I):

[0007]

[Chemical 12]

(In the formula, R is an alkyl group having 1 to 5 carbon atoms,
And n represents an integer of 4 to 22) to provide a carcinostatic agent containing a higher fatty acid as an active ingredient. The present invention also provides a cancer control enhancer containing the higher fatty acid as an active ingredient.

[0009]

EXAMPLES In the present invention, the chain length of the alkyl chain of the higher fatty acid is such that n in the general formula (I) is an integer of 4 to 22, and R 1 is an alkyl group having 1 to 5 carbon atoms. n
When is less than 4 and is greater than 22, it does not exert anti-tumor effect and anti-tumor enhancing effect. R is 6 carbon atoms
When it is an alkyl group as described above, it does not exert a carcinostatic effect or a carcinostatic effect. In the general formula (I), those having a particularly strong antitumor effect and antitumor enhancing effect are those in which the value of n is 9 to 14 and R is a methyl group or an ethyl group. Among them, formula (II):

[0010]

[Chemical 13]

12-methyltridecyl acid represented by the formula (I
II):

[0012]

[Chemical 14]

12-methyltetradecyl acid represented by the formula:
(IV):

[0014]

[Chemical 15]

14-methylpentadecyl acid represented by the formula: and formula (V):

[0016]

[Chemical 16]

14-Methylhexadecyl acid represented by the formula (1) shows particularly high carcinostatic action and antitumor enhancing action, and is particularly preferable.

The higher fatty acid used in the present invention is obtained by decomposing a natural product such as wax. Examples of natural products used include waxes such as wool wax, whale wax, beeswax, insect white wax, and carnauba wax.

The higher fatty acid 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 mixed with 1.18 times the molar amount of alkali (eg, NaOH).
A suspension is formed in water in the presence, and the saponification reaction is carried out at 135 ± 5 ° C under pressure for 3 hours with stirring (using an autoclave).

(Separation of fatty acid and alcohol) As a saponification reaction finished product (mixture of Na salt of higher fatty acid and higher alcohol)
Add H ₂ O and CH ₃ COC ₂ H ₅ (hereinafter MEK),
Transfer to a separatory funnel and heat to 70-75 ° C to remove alcohol from MEK.
Extract and separate into the inside. The higher fatty acid sodium salt in the aqueous layer is converted to free fatty acid with hydrochloric acid and extracted with MEK. The solvent is removed from the MEK solution of wool fatty acid under reduced pressure to obtain wool fatty acid as a solid.

(Molecular Distillation of Wool Fatty Acid) Solid wool fatty acid is subjected to molecular distillation to obtain a low boiling fraction (temperature: <80 ° C., pressure: 1
X10 ^-2 Torr). This fraction is designated as MD1-Acid.

(Fractionation of MD1-Acid by reverse phase column chromatography) Fractionation by MD1-Acid reverse phase column chromatography (open column) is performed for 5 fractions.

Fractions eluted from the third and fourth from the beginning are designated MD1-3-Acid and MD1-4-Acid, respectively.

These two fractions were each fractionated by high performance liquid chromatography (HPLC) and analyzed from MD1-3-Acid, for example, 12-methyltridecyl acid, 12-methyltetradecyl acid, MD1-4-Acid. The desired compound such as 14-methylpentadecyl acid or 14-methylhexadecyl acid is isolated.

Fractionation conditions Filler: Octyl group-modified silica gel, crushed, pore size 60
Angstrom, particle size 60/200 mesh (trade name: YM
C ・ GEL, manufactured by Yamamura Chemical Research Institute Co., Ltd. Eluent: CH ₃ OH / H ₂ O = 92/8 (volume ratio) In addition, higher fatty acids obtained in this way are, for example, R 2 being a methyl group. Sometimes, iso-C ₁₀ -COOH
(9-methyldecyl acid), iso-C ₁₁ -COOH (10-methyl undecyl acid), iso-C ₁₂ -COOH (11-methyl dodecyl acid), iso-C ₁₄ -COOH (13-methyl tetradecyl acid) ,
Examples include iso-C ₁₅ -COOH (14-methylpentadecyl acid), and when R is an ethyl group, iso-C ₁₂ -COO
H (10-methyldodecyl acid), iso-C ₁₃ -COOH (11-methyl tridecyl acid), iso-C ₁₅ -COOH (13-methyl pentadecyl acid), iso-C ₁₇ -COOH (15-methyl hepta) Decylic acid) and the like.

These higher fatty acids may be used alone or in admixture of two or more.

To prepare the carcinostatic agent and carcinostatic agent of the present invention for injection and infusion, Pluronic F-68 (trade name,
Asahi Denka Kogyo Co., Ltd., HCO-60 (trade name, Nikko Chemicals Co., Ltd.) and other surfactants are added and dispersed by ultrasonic waves, or made into liposomes or oil-in-water emulsions, and p-hydroxybenzoic acid Preservatives such as methyl acidate, stabilizers such as lecithin, linoleic acid, non-aqueous vehicles such as coconut oil, and suspending agents such as glucose can be included.

For oral preparation, a capsule suitable for intestinal absorption, a binder such as gelatin, a stabilizer such as magnesium stearate, an excipient such as lactose, and a disintegration such as potato starch. An enteric coating can be formed with a cellulose acetate phthalate, a methyl acrylate / methacrylic acid copolymer or the like by adding an agent. In addition, it can be formulated as granules, sustained-release buried capsules, suppositories, nebulizers, buccal agents.

The anticancer drug of the present invention is administered intravenously, subcutaneously,
For parenteral preparations such as infusion, the dose of active ingredient (adult body weight 1 kg, per day) 10 to 2000 mg, especially 50 to 600 m
g is preferred, and for oral administration agents such as capsules, 0.2-
70 g, especially 1-15 g is preferred. When used as a carcinostatic agent, the parenteral dose is 1.00 to 200 mg, preferably 3 to 50 mg, and the oral dose is 0.05 to 80 g, preferably 0.1 to 3.0 g.

The carcinostatic agent and carcinostatic agent of the present invention are:
It is effective against not only ascites cancer and leukemia but also solid cancer, and has a wide range of indications such as adenocarcinoma of each tissue, squamous cell carcinoma, undifferentiated cancer, and sarcoma. In addition, it is effective against not only cancer-transplanted animals but also cultured malignant cells such as humans, mice, rats, and hamsters, so that it has a direct cancer cell-killing effect and is not species-specific. It can be used as a cancer chemotherapy drug for animals.

Examples of the anticancer agent include, for example,
It can be used in combination with anticancer agents such as bleomycin, adriamycin, and vincristine.

Further, the carcinostatic agent and carcinostatic agent of the present invention have therapeutic effects not only by direct administration to the site of tumor implantation but also by remote administration. Toxicity LD ₅₀ is 8.0 to 25 g / kg by subcutaneous injection in rats, and no side effect is observed even after continuous administration of 1 to 2 g / kg for 10 days.

The production method and effectiveness of the higher fatty acid, which is the active ingredient of the anti-cancer agent and anti-tumor enhancer of the present invention, will be described below with reference to Production Examples and Test Examples. It is not limited to only.

Production Example 1 200 g of wool fatty acid obtained by saponifying wool grease was subjected to molecular distillation to obtain a low boiling fraction MD1-Acid (temperature:
<80 ° C., pressure: 1 × 10 ⁻² Torr) 26 g was obtained.

MD1-Acid was subjected to reverse phase column chromatography (open column), and mobile phase CH ₃ OH / H ₂ O: 92
Fractionation was performed at / 8 (volume ratio).

Of these, the third and fourth eluted fractions are referred to as MD1-3-Acid and MD1-4-Acid, respectively.

These MD1-3-Acid and MD1-4-Acid were fractionated by high performance liquid chromatography (HPLC) to isolate the desired compound.

From MD1-3-Acid, for example, 12-methyltridecyl acid and 12-methyltetradecyl acid, and from MD1-4-Acid, for example, 14-methylpentadecyl acid and 14-methylhexadecyl acid were obtained.

Then, it was confirmed by capillary gas chromatography that each higher fatty acid was clearly isolated.

¹³ C-NMR and GC-MS were used for the structure determination, and it was identified that the isolated substance was a higher fatty acid group represented by the general formula of the present invention.

Test Example 1 Five-week-old ddY mice were inoculated with 10 ⁶ Ehrlich's ascites tumor cells into the abdominal cavity, and 24 hours later, 0.25% (weight%, the same applies below) Pluronic F68 physiological saline solution was used to apply each sample. 50 mg
The liquid suspended at a concentration of 10 ml / ml is 10 animals per group, 5 mg at 10 mg / kg / day.
Injected ip daily.

The control group to which the same solution containing no sample was administered had an average survival time of 14.0 days after inoculation, while the group to which 12-methyltridecyl acid was administered had a survival time of 50 days or more, and the group to which 14-methylpentadecyl acid was It was 43 days, and the equivalent mixture of these was 45 days or more, showing a significant life prolonging effect.

Test Example 2 10 ⁶ adenocarcinoma 755 cells were subcutaneously transplanted to the back of 6-week-old mice of the first-generation hybrid of C57BL / 6 and DBA / 2, and 0.25% HCO-60 physiological saline solution was prepared 24 hours later. 50mg / each sample
The solution suspended in the concentration of ml was subcutaneously injected at 10 mg / kg / day for 5 days, with 8 animals per group. Adenocarcinoma 755 After transplantation of cells 10
On the day, mice were sacrificed and tumors excised.

The average tumor weight (g) was 6.3 in the control group, but was 2.5 and 3.1 in the groups administered with 12-methyltetradecyl acid and 14-methylhexadecyl acid, respectively, showing a significant tumor suppressive effect. .

Test Examples 3 and 4 and Comparative Test Example 1 In the case of using bleomycin alone and the case of using bleomycin and 12-methyltetradecyl acid or 14-methylhexadecyl acid in combination, mouse ascites tumor Ehrlich cells The cell killing effect (IC ₉₀ ) was examined using human lung cancer type A549 cells. The results are shown in Table 1.

[0047]

[Table 1]

As is clear from the results shown in Table 1, when the higher fatty acid of the present invention was used in combination with a carcinostatic agent, it was markedly killed compared with the case where the carcinostatic agent was used alone. It can be seen that the cell effect is improved.

Test Example 5 and Comparative Test Example 2 When bleomycin (60 μg / ml) and 10-methylundecyl acid (0.05 μM) were used in combination, the amount of bleomycin incorporated into Ehrlich cells was examined over time. . The results are shown in Table 2.

[0050]

[Table 2]

As is clear from the results shown in Table 2, when the higher fatty acid of the present invention is used in combination with a carcinostatic agent, the carcinostatic agent is more effective than the carcinostatic agent alone. It can be seen that the amount of the drug taken up into Ehrlich cells increases in a short time.

Formulation Example (Blending Ratio in Typical Dosage Form) Anticancer Agent (Bleomycin etc.) 0.005 to 100 mg / kg (Adult body weight 1 kg per day) Anticancer enhancer (higher fatty acid of the present invention) 1.00-200mg / kg (Adult body weight 1kg, per day) Excipients (lactose, crystalline cellulose, etc.) 10-99.8% (w / w) Lubricants (magnesium stearate, etc.) 0-50% (w / w )

[0053]

The carcinostatic agent and carcinostatic agent of the present invention exhibit excellent carcinostatic effect and carcinostatic effect,
Moreover, the active ingredient is derived from a living body (higher animal) and does not show serious side effects on the living body.

Continuation of front page (56) References JP-A-63-303925 (JP, A)

Claims (12)

[Claims] 1. A compound represented by the general formula (I): (In the formula, R is an alkyl group having 1 to 5 carbon atoms, and n is 4
A carcinostatic agent containing a higher fatty acid as an active ingredient. 2. The carcinostatic agent according to claim 1, wherein in the general formula (I), R is a methyl group or an ethyl group, and n is an integer of 9 to 14. 3. The active ingredient has the formula (II): The anticancer agent according to claim 1, which is 12-methyltridecyl acid represented by 4. The active ingredient has the formula (III): The anticancer agent according to claim 1, which is 12-methyltetradecyl acid represented by 5. The active ingredient has the formula (IV): The anticancer agent according to claim 1, which is 14-methylpentadecyl acid represented by 6. The active ingredient has the formula (V): The anticancer agent according to claim 1, which is 14-methylhexadecyl acid represented by 7. A compound represented by the general formula (I): (In the formula, R is an alkyl group having 1 to 5 carbon atoms, and n is 4
~ Represents an integer of 22), a carcinostatic agent comprising a higher fatty acid as an active ingredient. 8. The antitumor agent according to claim 7, wherein in the general formula (I), R is a methyl group or an ethyl group, and n is an integer of 9 to 14. 9. The active ingredient has the formula (II): The anticancer enhancer according to claim 7, which is 12-methyltridecyl acid represented by 10. The active ingredient has the formula (III): The anticancer enhancer according to claim 7, which is 12-methyltetradecyl acid represented by 11. The active ingredient has the formula (IV): The antitumor agent according to claim 7, which is 14-methylpentadecyl acid represented by 12. The active ingredient has the formula (V): The anticancer enhancer according to claim 7, which is 14-methylhexadecyl acid represented by