JPH10259171A - Solvent extraction of taxane compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To accomplish safe and efficient solvent extraction of a taxane compound in a short time by bringing a taxane compound-contg. mixture into contact with a solvent capable of dissolving at least part of the taxane compound to effect extraction under heating and pressure. SOLUTION: A mixture containing at least one kind of taxane compound selected from taxol, cephalomannine, baccatin III, 10-deacetylbaccation III and 10-deacetylcephalomannine (pref. a plant such as a tree belonging to the genus Taxus, or oil extracted therefrom) is brought into contact with a solvent capable of dissolving at least part of the above taxane compound(s), for example, organic solvent, water or a mixture thereof, pref. methanol, water or a mixture of methanol and water at 9:1 in volume ratio [normally, in an amount of 3-50 (pref. 5-20)wt. times the mixture containing the taxane compounds(s)] to effect the objective extraction under 10-200atm at 40-190 deg.C and normally for 15-20min or so by use of pref. a high-performance solvent extractor.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a taxane compound (ta
xanes)
on). More specifically, the present invention relates to a solvent extraction method for taxol, which is known as an anticancer agent, and a taxane compound useful as an intermediate for producing taxol.

[0002]

2. Description of the Related Art Taxane compounds are diterpene compounds known to be used in the pharmaceutical field. Among them, taxol is an effective anticancer agent. Taxane compounds are contained in plants, but various taxane compounds contained in plants are trace amounts. Therefore, the ordinary liquid extraction method requires a long time and requires a large amount of the organic solvent, so that there is a concern that the solvent may be deteriorated. When water is used as a solvent instead of an organic solvent, it is difficult to extract under normal pressure. Therefore, there is a need for a safe and efficient extraction method in a short time.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a solvent extraction method for a taxane compound which can be safely and efficiently extracted in a short time. is there.

[0004]

The gist of the present invention is to provide (1)
A method for extracting a taxane compound from a mixture containing a taxane compound, comprising contacting the mixture with a solvent that dissolves at least a portion of the taxane compound, and extracting the mixture under heating and pressure. (2) wherein the taxane compound to be extracted is at least one selected from the group consisting of taxol, cephalomanine, baccatin III, 10-deacetylbaccatin III and 10-deacetylcephalomanine; (3) the extraction method according to (1), wherein the taxane compound to be extracted is taxol;
(4) The extraction method according to any one of (1) to (3), wherein the mixture containing the taxane compound is a plant or an extracted oil thereof, (5) the extraction according to (4), wherein the plant is a tree of the genus Taxus. Method, (6) the tree of the genus Taxus is T.
Casvidata, T.M. Brevifolia, T. Baccata,
T. Media, T. Warichiana and T.M. (5) the extraction method according to (5), wherein the plant is at least one selected from the group consisting of Canadensis;
The extraction method according to any one of the above (4) to (6), which has at least a part selected from the group consisting of branches, woody parts, seeds and whole seedlings, (8) the solvent is an organic solvent, water, or a mixture thereof. (9) 1 The extraction method according to (1) above,
The extraction method according to the above (1), wherein the pressure is increased to 0 to 200 atm;
(10) The extraction method according to (1), wherein the heating temperature at the time of extraction is equal to or lower than the critical temperature of the solvent. (11) The (10) wherein the heating temperature at the time of extraction is equal to or higher than 40 ° C. and equal to or lower than the critical temperature of the solvent. (12) The extraction method according to (11), wherein the heating temperature during the extraction is 70 to 190 ° C.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION In the solvent extraction method for a taxane compound of the present invention, a mixture containing the taxane compound is brought into contact with a solvent in which at least a part of the taxane compound is dissolved.
It is a method characterized by extracting under heat and pressure.

The taxane compound extracted according to the present invention is a diterpene compound having a taxane carbon skeleton, wherein the carbon skeleton contains an ethylenically unsaturated group in its ring system.

The taxane compounds include taxol, cephalomannine, baccatin III
(Baccatin III), 10-deacetylbaccatin III
(10-Deacetyl baccatin III) and 10-Deacetyl Cephlomannine
It is preferable that at least one selected from the group consisting of

[0008] In the present invention, the mixture containing the taxane compound is preferably a plant or its extracted oil.

In the present invention, a typical example of a plant to be used is a tree of the genus Taxus.

[0010] As the tree of the taxus genus, for example,
T. Casvidata, T.M. Brevifolia, T. Baccata, T. Media, T. Warichiana, T .; Canadengis, T .; Sumatrana and the like. Among these,
T. Casvidata, T.M. Brevifolia, T. Baccata, T. Media, T. Warichiana and T.M. It is preferably at least one member selected from the group consisting of Canadensis, and in particular, T. canadensis is easily available in Japan. Casvidata is preferred.

In the present invention, the plant is bark,
It is preferable to have at least a part selected from the group consisting of roots, leaves, branches, xylem, seeds and whole seedlings (seedlings). Prior to contact with the solvent, it is preferable to use a plant obtained by crushing the plant to obtain a suitable grain (10 to 2000 μm).

The solvent used in the present invention may be any solvent that dissolves at least a part of various taxane compounds, and is not particularly limited as long as the solvent can dissolve the taxane compound targeted for extraction. When selecting a solvent, a solvent having a higher solubility of the taxane compound is preferable from the viewpoint of extraction efficiency.

As the solvent, for example, an organic solvent,
Water or a mixture thereof is included. Examples of such organic solvents include alcohols such as methanol, ethanol, and isopropanol; esters such as ethyl acetate; chloroform; chloroalkanes such as methylene chloride; ketones such as acetone; diethyl ether; ethers such as tetrahydrofuran; Examples include aromatic hydrocarbons such as benzene and toluene, and nitrogen compounds such as acetonitrile. Among these solvents, methanol, water, or a mixture of methanol and water (for example, methanol: water = 9: 1 (volume ratio)) is preferable from the viewpoint of the ability to extract the target component (taxane compound). When the taxane compound targeted for extraction is provided for medical use,
It is preferable that a pharmaceutically acceptable solvent is appropriately selected.

The amount of the solvent used is usually 3 to 50 times by weight, preferably 5 to 50 times the weight of the mixture containing the taxane compound.
Desirably, it is 20 times by weight.

In the present invention, the pressure at the time of pressurization depends on the type of the solvent, the extraction temperature, and the like.
0 atm or more, preferably 50 atm or more, and 200 atm or less from the viewpoint of the pressure resistance of the device.
Preferably, it is desirable that it be 150 atm or less.

In the present invention, the mixture containing the taxane compound is brought into contact with the above-mentioned solvent, and the extraction operation is performed under heating and pressurization. In the extraction, the solvent is preferably used in a state that does not become a supercritical fluid. Therefore, the heating temperature at the time of extraction is preferably equal to or lower than the critical temperature of the solvent. Specifically, the heating temperature at the time of extraction is preferably 40 ° C. or higher, and more preferably 70 ° C. or higher for the purpose of increasing the extraction efficiency, and 190 ° C. or lower for suppressing the thermal decomposition of the target component (taxane compound). , Preferably 150 ° C. or lower.

The extraction method of the present invention is a high-speed solvent extraction method which can usually extract in a short time of about 15 to 20 minutes.

The extraction method of the present invention is preferably carried out using a high-speed solvent extraction apparatus. For example, ASE-200 (manufactured by Nippon Dionex Co., Ltd.), an apparatus having the same function, or the like can be used.

The taxane compound obtained by the solvent extraction method of the present invention is useful in the field of medicine, particularly as an anticancer agent. Especially, taxol is known as a taxane compound having an anticancer effect. Other taxane compounds are also useful as intermediates of drugs having other pharmacological actions.

[0020]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the present invention. The extraction ratio (%) indicates the weight% of the extracted component with respect to 5 g of bark.

Example 1 Japanese yew (T. cuspidata) bark (collected from a 16th year old tree in Monbetsu, Hokkaido) was pulverized with a pulverizer (Retsch, cutting mill SM11) to a particle size of 2 mm or less, and 5 g of the pulverized material was used. Take methanol 40 as solvent
together with the solvent in a high-speed solvent extraction apparatus (Model ASE-200, manufactured by Nippon Dionex Co., Ltd.).
The extraction operation was performed for 5 minutes. After the extraction, the taxane compound (taxol, baccatin III, 10-deacetylbaccatin III (hereinafter, 1)) contained in the solution taken out is subjected to high-performance liquid chromatography using the following high-performance liquid chromatograph and measurement conditions.
0-DAB))), and the extraction ratio was measured. Table 1 shows the results.

[High-performance liquid chromatograph]-Shimadzu high-performance liquid chromatograph (HPLC)-Pump: LC-10AD-Detector: SPD-M10A-Column oven: OTO-10A-Column: Senshu Kagaku, PEGASIL C8

[Measurement conditions] Eluent: acetonitrile: 10 mM ammonium acetate water = 45: 55 (volume ratio) Flow rate: 1.0 ml / min Detection: UV 228 nm Column temperature: 40 ° C.

The amounts of the extracted components were determined by using standard taxol (trade name: Paclitaxel), baccatin III (trade name: BACCATIN III) and 10-DAB, which were sold by SIGMA.
(Product name: 10-DEACETYL BACCATIN III) was performed according to a calibration curve created.

[0025]

[Table 1]

From Table 1, it can be seen that when methanol was used as a solvent and the extraction was performed at 100 atm, the taxol extraction rate was 1%.
It can be seen that taxol was extracted at the highest at 50 ° C. and 1.48 times that at 40 ° C.

Example 2 The extraction rate of a taxane compound was measured in the same manner as in Example 1 except that the temperature was changed to 150 ° C. and the pressure was changed to each pressure shown in Table 2. Table 2 shows the results.

[0028]

[Table 2]

According to Table 2, the extraction rate of taxol at 150 ° C. is the highest at 100 atm, but good results are seen at 10 atm.

Example 3 Instead of methanol, each solvent shown in Table 3 was used, and the temperature condition was 100 ° C. and the pressure condition was 100 at.
The extraction rate of the taxane compound was measured in the same manner as in Example 1 except that the value was changed to m. Table 3 shows the results.

[0031]

[Table 3]

From Table 3, it can be seen that various solvents are used at 100 ° C. and 1
When extracted at 00 atm, the extraction ratio of taxol in the case of methanol showed a maximum value of 0.087%, but in the case of water, it was found that the extraction ratio was relatively high at 0.0604%.

Example 4 Using a mixed solvent of methanol and water in the respective mixing ratios shown in Table 4, the temperature condition was 150 ° C., and the pressure condition was 100 a.
Except for tm, the extraction rate of the taxane compound was measured in the same manner as in Example 1. Table 4 shows the results.

[0034]

[Table 4]

According to Table 4, methanol: water = 90: 10.
At the time of (volume ratio), the highest taxol extraction rate (0.1279%) so far was obtained.

Comparative Example 1 A taxane compound was extracted in the same manner as in Example 1 except that the temperature condition was 24 ° C. and the pressure condition was 100 atm, and the extraction rate was measured. Table 5 shows the results.

[0037]

[Table 5]

From Table 5, the extraction rate of the taxane compound was lower than the result obtained in Example 1.

Comparative Example 2 A taxane compound was extracted in the same manner as in Example 1 except that the temperature condition was 64 ° C. and the pressure condition was 1.0 atm, and the extraction rate was measured. Table 6 shows the results.

[0040]

[Table 6]

As shown in Table 6, the extraction rate of the taxane compound was lower than the result obtained in Example 1.

Comparative Example 3 5 g of yew bark similar to that used in Example 1 was placed in a 100 cc stoppered Erlenmeyer flask containing 60 ml of each solvent shown in Table 7, and immersed for 48 hours at room temperature and atmospheric pressure. Thereafter, after filtration, the residue was washed with 60 ml of the same solvent as the solvent used for the immersion, and the taxane compound was extracted by concentrating together with the filtrate previously filtered, using a high-performance liquid chromatograph. In the same manner as in Example 1, the extraction rate of the taxol compound was measured. Table 7 shows the results.

[0043]

[Table 7]

As shown in Table 7, at room temperature and atmospheric pressure, the mixed solvent of methanol and methylene chloride has the highest taxol extraction rate, but it took 48 hours for extraction. On the other hand, in the case of water, 1
In the case of 00 ° C and 100 atm (extraction rate of taxol.
0604%) and as low as 0.0016%. This indicates that water has a good extraction rate of taxol only at high temperature and high pressure.

[0045]

According to the solvent extraction method for a taxane compound of the present invention, the taxane compound can be safely and efficiently extracted in a short time.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Tatsuro Ohira 3312-2 Ushikucho, Ushiku City, Ibaraki Pref. 3-14-10 Kiyomidai, Kawachinagano City, Osaka Prefecture

Claims (12)

[Claims] 1. A method for extracting a taxane compound from a mixture containing the taxane compound, the method comprising:
A solvent extraction method for a taxane compound, comprising bringing the taxane compound into contact with a solvent in which at least a part of the taxane compound is dissolved, and extracting the mixture under heat and pressure. 2. The extraction according to claim 1, wherein the taxane compound to be extracted is at least one selected from the group consisting of taxol, cephalomannine, baccatin III, 10-deacetylbaccatin III and 10-deacetylcephalomanine. Law. 3. The method according to claim 1, wherein the taxane compound to be extracted is taxol. 4. The extraction method according to claim 1, wherein the mixture containing the taxane compound is a plant or its extracted oil. 5. The plant according to claim 4, wherein the plant is a tree of the genus Taxus.
Extraction method as described. 6. The tree of the taxus genus T. Casvidata,
T. Brevifolia, T. Baccata, T. media,
T. Warichiana and T.M. The extraction method according to claim 5, wherein the method is at least one selected from the group consisting of Canadensis. 7. The extraction method according to claim 4, wherein the plant has at least a part selected from the group consisting of bark, roots, leaves, branches, woody parts, seeds and whole seedlings. 8. The method according to claim 1, wherein the solvent is an organic solvent, water, or a mixture thereof. 9. The method according to claim 1, wherein the pressure is increased to 10 to 200 atm.
Extraction method as described. 10. The extraction method according to claim 1, wherein the heating temperature during the extraction is lower than the critical temperature of the solvent. 11. The extraction method according to claim 10, wherein the heating temperature at the time of extraction is 40 ° C. or higher and not higher than the critical temperature of the solvent. 12. The extraction method according to claim 11, wherein the heating temperature during the extraction is 70 to 190 ° C.