WO1997024115A1

WO1997024115A1 - Naphthalenecarboxylic acid derivatives

Info

Publication number: WO1997024115A1
Application number: PCT/JP1996/003859
Authority: WO
Inventors: Tetsuya Matsuno; Yasuyuki Matsumoto; Junji Morikawa; Masahiro Saito
Original assignee: Eiken Kagaku Kabushiki Kaisha
Priority date: 1995-12-29
Filing date: 1996-12-27
Publication date: 1997-07-10
Also published as: JPH09183724A

Abstract

Antitumor agents containing as the active ingredient 1,2,3,4,4a,7,8,8a-octahydro-1,4a,5-trimethyl-1-(3-methyl-5-oxo-3-pentenyl)-2-naphthalenecarboxylic acids represented by chemical formula (I) or isomers or salts thereof.

Description

Description Naphthalene carboxylic acid derivative Technical field

The present invention relates to a compound of the formula (I)

1,2,3,4,4a, 7,8,8a-octyhydro-1, 4a, 5-trimethyl-11- (3-methyl-5-oxo-1-3-pentenyl) 1. 2-Naphthalenecarboxylic acid (hereinafter abbreviated as naphthylene carboxylic acid derivative) as a drug, particularly as an antitumor agent. Background art

Antineoplastic agents used to treat cancer can be broadly classified into chemotherapeutic agents and immunotherapy agents.

Chemotherapeutic agents are used not only for unresectable cancers, but also in combination with surgery, given before and after surgery. For example, cancer chemotherapeutic agents include alkylating agents (nitrogen mass compounds, ethyleneimines, sulfonic esters, etc.), antimetabolites (folate antagonists, pyrimidine antagonists, etc.), plant nuclear fission Toxins (colcemid, vinblastine, etc.), antibiotics (sarcomycin, carcinophilin, mitomycin, etc.), hormonal drugs (adrenocortical steroids, male hormones) And porphyrin complex (morphylin, copp) etc. are used clinically.

However, chemotherapeutic agents are often cytotoxic substances and not only attack cancer cells, but also act on normal cells, and generally have strong side effects. For example, side effects such as vomiting, nausea, anorexia, malaise, neuropathy, myelopathy (leukopenia), hair loss, and stomatitis may be seen. Therefore, caution is required for long-term administration. On the other hand, immunotherapeutic agents are mainly immunostimulants, and increase the immunity to make immune response cells recognize cancer cells as foreign substances and treat them. However, immunostimulants have few side effects, but generally have mild antitumor effects and are often used as adjuvant therapy.

As described above, conventional antitumor agents have advantages and disadvantages. Therefore, development of an antitumor agent which has high antitumor activity, has few side effects, and can be administered for a long time has been desired. In addition, since antitumor drugs are generally expensive, it is also desirable that they be supplied at low cost to reduce the economic burden on patients.

Propolis, which is used as a folk remedy, is a resinous substance that is a mixture of vegetation collected by bees and saliva, honey, pollen, etc., and has a pharmacological action such as antibacterial action and anti-inflammatory action. It has been known. It is also known that drinking propolis as a health supplement has an antitumor effect, and an attempt has been made to extract and purify an antitumor active substance from propolis (Japanese Patent Application Laid-Open No. 5-58943). However, the extract has been found to have antibacterial activity and antitumor activity. Disclosure of the invention

Means for Solving the Problems The present inventors have intensively studied to find a physiologically active substance having an antitumor activity from propolis, and as a substance having an antitumor activity, the following chemical formula (I)

A naphthene carboxylic acid derivative represented by the formula was found.

The naphthenic carboxylic acid derivative represented by the chemical formula (I) is a plant of the species Symphyopappus (PHYTOCHEMISTRY, 20 (7), 1657-1663, 1981) ゃ Zenigoke (PHYTOCHEMISTRY, 28 (12) , 3415-3419, 1989). In addition, the following chemical formulas (I I) and (I I I)

)

Is registered in the Chemical Abstracts (CHEMICAL ABSTRACT) as registration numbers 126239-179-0 and 1 239239-80-3. However, it is unknown what kind of pharmacological action this compound has or whether it can be used as a medicament, and it is a new finding by the present inventors that these compounds have an antitumor effect.

Specifically, the present invention provides a pharmaceutical composition comprising a naphthylene carboxylic acid derivative represented by the above formula (I) as an active ingredient, particularly a pharmaceutical composition for treating or preventing a tumor. .

That is, the present invention

[1] A pharmaceutical composition comprising, as an active ingredient, a naphthylene carboxylic acid derivative represented by the chemical formula (I) or a salt thereof.

[2] The pharmaceutical composition according to [1], wherein the stereostructure of the naphthylene carboxylic acid derivative is represented by the above chemical formula (II) or (III).

[3] A pharmaceutical composition for treating or preventing tumors, comprising a naphthylene carboxylic acid derivative represented by the chemical formula (I) or a salt thereof as an active ingredient.

[4] The pharmaceutical composition according to [3], wherein the three-dimensional structure of the naphthylene carboxylic acid derivative is represented by the above chemical formula (II) or (III).

[5] use of a naphthalene carboxylic acid derivative represented by the above formula (I) or a salt thereof for the preparation of a pharmaceutical composition for treating or preventing a tumor;

[6] The use according to [5], wherein the stereostructure of the naphthylene carboxylic acid derivative is represented by the above chemical formula (II) or (III).

[7] a method for treating or preventing a tumor, which comprises administering a composition comprising a naphthalenecarboxylic acid derivative represented by the chemical formula (I) or a salt thereof; and

[8] The method according to [7], wherein the three-dimensional structure of the naphthalenecarboxylic acid derivative is represented by the above chemical formula (II) or (III).

About. The salt of the naphthylene carboxylic acid derivative of the present invention includes pharmaceutically acceptable salts.

Since the naphthenic carboxylic acid derivative of the present invention has an antitumor cell effect on tumor cells as shown in the examples, it is effective to administer it in various modes to treat or prevent tumors. It is considered to be effective. The method of administering the compound may be parenteral or oral, and the composition administered may include a therapeutically effective amount of the compound and a pharmacologically acceptable diluent, stabilizer, excipient, etc. Is contained. Examples of the administration form include intravenous injection, subcutaneous injection, intramuscular injection, parenteral administration such as suppository, soft and tincture, and oral administration using tablets, powders, capsules, granules and the like.

Since the naphthylene carboxylic acid derivative of the present invention is obtained as an oily substance or a crystalline powder, when administered in a dry form such as a tablet or powder, it is preferable to use the isomer on the powder, and to administer the liquid as a liquid. If used, they may be dissolved or suspended in oils or powders and used if appropriate for the dosage form. In the case of a form such as a double powder, even if it is oily, it can be used as a powder.

Oral administration of propolis itself to dogs, rats and guinea pigs at doses of 10 to 15 g / kg for several months showed no toxicity (PROPOL IS: 2nd ed., Y. D0NAD I EU Therefore, it is considered that the safety of administering naphthene carboxylic acid derivatives to humans is high.

In addition, alcoholic extracts of propolis are known to have antibacterial, antioxidant, anti-inflammatory, virus growth inhibitory, macrophage activating, hair-growth effects, etc. No. 316968, JP-A-7-298833, etc.), and the naphthylene carboxylic acid derivative of the present invention obtained by purifying an alcoholic extract of propolis can also be expected to have these effects. BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a diagram showing a mass spectrum of a naphthene carboxylic acid derivative (II).

FIG. 2 is a diagram showing a NMR spectrum of the naphthylene carboxylic acid derivative (II).

FIG. 3 is a diagram showing a mass spectrum of a naphthene carboxylic acid derivative (III).

FIG. 4 is a diagram showing a NMR spectrum of a naphthylene carboxylic acid derivative (III). BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in more detail with reference to Examples. The following examples are merely for explanation, and do not limit the present invention in any way.

Example 1 Extraction of the present compound from propolis

Extraction of Naphthylene Carboxylic Acid Derivative from Propolis' Purification was performed through the following [Step 1] to [Step 5].

[Step 1]

100 g of propolis was mixed with 10 times the amount of 99.5% ethanol, and the mixture was stirred at room temperature using a magnetic stirrer. The suspension obtained was filtered under reduced pressure to obtain an ethanol extract of propolis. Then, the solvent of the obtained ethanol extract was removed by evaporation using a rotary evaporator overnight, about 30 g of the obtained residue was dispersed in a mixture of water and ethyl acetate (1: 1), and the upper ethyl acetate layer was separated. An ethyl acetate extract of propolis was obtained. The solvent of the obtained ethyl acetate extract was evaporated and removed on a rotary evaporator, and about 20 g of the obtained residue was dispersed in 99.5% methanol.Insoluble matters were removed by low-speed centrifugation, and methanol extraction of propolis was performed. A liquid was obtained.

[Step 2]

High-performance liquid obtained by equilibrating the methanol extract obtained in step 1 with 70% methanol The mixture was injected into an ODS column for chromatography and an ODS 80TM column (manufactured by Tosoh Corporation), and gradient elution was performed with a methanol concentration gradient of 70% to 100%. A fraction having a methanol concentration of 90 to 95% was collected.

[Step 3]

The fractions having a methanol concentration of 90 to 95% obtained in Step 2 were collected into one, and the solvent was removed by evaporation on a rotary evaporator to obtain about 2 g of a residue. Next, the obtained residue is dissolved in a gel form, and the obtained solution is injected into an Inertsil series column for high-performance liquid chromatography and an Inertsil SIL column (manufactured by GE Science) equilibrated with the gel form and retained. Two fractions with different times were collected.

[Step 4]

The two fractions of the solvent obtained in step 3 were evaporated off on a rotary evaporator to obtain about 70 mg and 60 mg of the residue, respectively. Next, each of the obtained residues was dissolved in a chromate form, and the resulting solution was injected into a GPC column for high performance liquid chromatography and a Shodex GPC-H2000 column (manufactured by Showa Denko KK) equilibrated with the chromate form. Then, the main fraction was fractionated.

[Step 5]

The solvent of each main fraction obtained in step 4 was removed by evaporation using a rotary evaporator to obtain about 50 mg and 40 mg of a residue as colorless crystalline substances, respectively. From the physicochemical properties of the residues, they were 1,2,3,4,4a, 7,8,8a—Hydrohydro-1,4a, 5-trimethyl-1- (3-methyl-5-oxo-3-) (Pentenyl) isomers of 2-naphthylene carboxylic acid (II) and (III).

Further, in step 3, the isomers were not separated, and the subsequent operation was performed in the same manner to obtain compound (I). Compound (I) was obtained as an oil.

Example 2 Physicochemical properties of naphthalene carboxylic acid derivative (II)

Compound name: 2-Naphtha 1 enec arbox 1 ic acid, l, 2,3,4,4a, 7,8,8a-octahydro-l, 4 a, 5-trimethyl-l- (3-methyl-5-oxo-3-pentenyl)-, [lR- [l.alpha. (Z), 2.beta., 4 a.beta., 8a.alpha. ]]-(9CI) (Chemical Abstract (CA) Index Name) Chemical Abstract (CA) Registration No .: 126239-79-0

Molecular weight: 318.46

Molecule ί¾: C20H3O 3

Chemical formula:

Appearance: Colorless needle-like crystals or crystalline powder, odor is aromatic.

Solubility: Easily soluble in chloroform and dimethylformamide, ethanol and methanol

Soluble in water, insoluble in water and cyclohexane.

Mass spectrum:

Equipment: Finnigan MAT I NCOS 50

Ionization: EI

Acceleration port number: 70 eV

The results are shown in Figure 1.

NMR spectrum:

JEOL JNM-GSX400

Resonance Ή (400 MHz)

Sambruhase 7.7 mg / 0.55 ml CDCh Internal control: TMS

Temperature: 30. C

The result is shown in figure 2.

HP L C:

Equipment: SHIMADZU CL-4A

Separation conditions and retention times are as follows, Table 1.

From the analysis of these data, the naphthylene carboxylic acid derivative of the present invention was identified as the compound represented by the chemical formula (II).

Example 3 Physicochemical properties of naphthalenecarboxylic acid derivative (II)

Name of compound: 2-Naphthalenecarboxyl ic acid, 1,2,3,4, a, 7,8,8a-octahydro-l, 4a, 5-tr imethy 1 -1-(3-methyl -5- oxo-3-penten 1)-, [1R- [1. alpha. (E), 2. beta., 4 a. beta., 8a. alpha.]]-(9CI) (Chemical Abstract (CA) Index name) Chemical Abstracts (CA) Registration No. 126239-80-3

Molecular weight: 3 1 8.46

Molecular formula: C _2. H _3. 0 ₃

Chemical formula:

Soluble in water, insoluble in water and cyclohexane.

Mass spectrum:

Equipment: Finnigan MAT INCOS 50

Ionization: EI

Acceleration volts: 70 eV

The results are shown in Figure 3.

NMR spectrum:

JEOL JNM-GSX400

Resonance 1H (400 MHz)

Samburuzu: 4.05 mg / 0.55 ml CDC 13

Internal control TMS

Temperature 25. C

The results are shown in FIG. HP LC:

Equipment: SHIMADZU CL-4A

Separation conditions and retention times are as follows. Table 2

From these analyzes, the naphthene carboxylic acid derivative of the present invention was identified as the compound represented by the chemical formula (III).

Example 4 Antitumor cell action on cultured tumor cells

Using the naphthalenecarboxylic acid derivatives (I), (II) and (III) obtained in Example 1 as test substances, a cell damage activity test of tumor cells was performed as follows.

To each well of a 96-well microtiter plate, 0.1 ml of a test substance appropriately diluted with a MEM medium containing 10% fetal calf serum and 2 mM glucumin was added, and then a human liver cancer HuH treated with tribcine was added. 13 shares (J. CELLULAR PHYSYOL., VOL 148, 290-294, 1991) was adjusted to 3 × 10 ⁴ cells / ml with the above culture solution, and dispensed in 0.05 ml portions.

After culturing the plate in a carbon dioxide incubator at 37 ° C for 72 hours, the culture supernatant was removed, and 0.1 ml of a culture solution containing 0.02% neutral red was added to each well at 37 ° C. The cells were cultured in a carbon dioxide incubator for 1 hour, and the cells were stained. After removing the culture supernatant, the residue was washed once with physiological saline. Next, the dye was extracted with 0.001N hydrochloric acid / 30% ethanol, and the absorbance at 550 nm was measured using a microplate reader. The absorbance of untreated cells was compared with that of cells treated with a known concentration of the test substance, and the cell growth inhibition rate was calculated according to the following equation. Growth inhibition rate (%) = 100-[(A-B) / (C-B)] X 100

A: Absorbance of drug-treated cells

B: Absorbance of cell-free well

C: Absorbance of untreated cells From the obtained growth inhibition rate, the concentration of a test substance that inhibits cell growth by 50% (ID ₅₀ ) was calculated. Table 3 shows the results. Table 3

Test tumor cells

Compound (I) Compound (II) Compound (III)

HuH 1 3 shares 30 30 30 As is evident from Table 3, the naphthylene carboxylic acid derivatives (I), (II) and (III) exhibited excellent growth inhibitory effects on tumor cells. Industrial applicability

The naphthylene carboxylic acid derivative (1, 2, 3, 4, 4a, 7, 8, 8a-octahydro-1,4-a, 5-trimethyl_1) of the present invention which is easily obtained by extraction from propolis Mono (3-methyl-5-oxo_3-pentenyl) -1-naphthylene carboxylic acid and its isomers exhibit antitumor activity and are useful as antitumor agents. It is expected to be an antitumor agent with high safety and few side effects.

Claims

The scope of the claims

1. The following chemical formula (I)

A pharmaceutical composition comprising, as an active ingredient, a naphthalene carboxylic acid derivative represented by the formula: or a salt thereof.

2. The three-dimensional structure of the naphthalenecarboxylic acid derivative has the following chemical formula (II)

Or chemical formula (III)

The pharmaceutical composition according to claim 1, represented by:

3. A pharmaceutical composition for treating or preventing a tumor, comprising a naphthylene carboxylic acid derivative represented by the above formula (I) or a salt thereof as an active ingredient.

4. The pharmaceutical composition according to claim 3, wherein the three-dimensional structure of the naphthylene carboxylic acid derivative is represented by the chemical formula (II) or (III).

5. Use of a naphthylene carboxylic acid derivative represented by the above formula (I) or a salt thereof for the preparation of a pharmaceutical composition for treating or preventing a tumor.

6. The use according to claim 5, wherein the stereostructure of the naphthylene carboxylic acid derivative is represented by the chemical formula (II) or (III).

7. A method for treating or preventing a tumor, comprising administering a composition comprising a naphthylene carboxylic acid derivative represented by the above formula (I) or a salt thereof.

8. The method according to claim 7, wherein the stereostructure of the naphthylene carboxylic acid derivative is represented by the above chemical formula (II) or (III).