JP6409415B2 - Method for producing atractylodins and cobalt complex compound - Google Patents

Description

  TECHNICAL FIELD The present invention relates to a method for producing atractyrosines and a cobalt complex compound, and more particularly to a method for producing atractyrosines using a cobalt complex compound of atractyrosin that can be stored for a long period of time, and the cobalt complex compound.

  Attractyrosin is a polyene compound having a characteristic diene-diyne structure, and is a major component of the herbal medicine “Sojitsu”. It is also known as the main medicinal ingredient of blended Kampo prescriptions, including Rikkunshito, which uses herbal medicine as a constituent crude drug. In recent years, an interesting pharmacological mechanism has been clarified such that atractyrosin has an appetite enhancing action through ghrelin regulation, and has attracted more and more attention as an important component in clinical application and quality control.

  Non-Patent Document 1 describes a method for synthesizing atractylodin. It is also possible to extract atractyrosin from natural products. For example, Non-Patent Document 2 discloses that atractyrosin is extracted from the essential oil components of Sojitsu by combining a supercritical fluid extraction (SFE) system and molecular distillation. How to do is described. Patent Document 1 also describes a method of purifying by extracting atractyrosin from Sojitsu and then recrystallizing it.

JP 59-106421 A

Annabelle L.K .; Shi Shun; Rik R. Tykwinski J. Org. Chem., 68, 6810-6813, (2003) Gao, Y .; et al .; Guangzhou Zhongyiyao Daxue Xuebao, 21, 6, 476-478, (2004)

  However, since atractyrosin has an unstable chemical structure, it has poor solid stability and is difficult to store for a long time because it decomposes with light or oxygen.

  Accordingly, an object of the present invention is to provide a method for producing atractylodins using a cobalt complex compound of atractyrosin capable of long-term storage, and the cobalt complex compound.

  As a result of intensive studies to solve the above problems, the present inventors have found that the above problems can be solved by using a specific novel cobalt complex compound, and have completed the present invention.

  The manufacturing method of the cobalt complex compound and atractyrosin of the present invention is the following [1] to [5].

（式中、ＡはＣＯを表し、Ｒ^１は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。） [1] A cobalt complex compound represented by the following general formula (I):

(In the formula, A represents CO, and R ¹ represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group, or a carboxyl group.)

（式中、Ｒ^２は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。）
で表されるアトラクチロジン類と、
下記一般式（ＩＩＩ）、

（式中、ＡはＣＯを表す。）
で表されるコバルト化合物とを反応させる工程を備える［１］のコバルト錯体化合物の製造方法。 [2] The following general formula (II),

(In the formula, R ² represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
Atractylodins represented by
The following general formula (III),

(In the formula, A table to the CO.)
The manufacturing method of the cobalt complex compound of [1] provided with the process with which the cobalt compound represented by these is made to react.

（式中、ＡはＣＯを表し、Ｒ^１は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。）
で表されるコバルト錯体化合物と脱保護剤とを反応させる工程を備えることを特徴とするアトラクチロジン類の製造方法。 [3] The following general formula (I),

(In the formula, A represents CO, and R ¹ represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group, or a carboxyl group.)
A process for reacting a cobalt complex compound represented by the formula: and a deprotecting agent.

[4] The method for producing atractyrosines according to [3], wherein the deprotecting agent is selected from the group consisting of ethylenediamine, diethylenetriamine, triethylenetetramine, and iron (III) nitrate.

（式中、Ｒ^２は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。）
で表されるアトラクチロジン類を昇華して精製する工程を含む［３］または［４］のアトラクチロジン類の製造方法。 [5] The following general formula (II),

(In the formula, R ² represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
[3] or [4] The method for producing atracty rosin comprising the step of sublimating and purifying the atracty rosin represented by the formula:

  INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a method for producing atractylodins using a cobalt complex compound of atractyrosin that can be stored for a long period of time, and the cobalt complex compound.

2 is a ¹ H-NMR spectrum of a cobalt complex compound of atractyrosin obtained in Experimental Example 1. FIG. ^{1 is} a ¹ H-NMR spectrum of atractyrozine obtained in Experimental Example 2-1. 6 is a photographic diagram showing the results of TLC analysis for stability evaluation of Experimental Example 3. FIG. FIG. 10 is a schematic diagram showing an apparatus used in Experimental Example 5.

  Hereinafter, embodiments of the present invention will be described in detail.

（式中、ＡはそれぞれＣＯ、Ｐ（Ｒ^Ａ）_３またはＮＯを表し、互いに同一でも異なっていてもよく、Ｒ^Ａは水酸基、アルデヒド基、カルボキシル基またはエステル基で置換されてもよい炭素数１〜４の直鎖または分岐鎖のアルキル基；フェニル基；アルデヒド基；カルボキシル基またはエステル基を表し、Ｒ^Ａが２以上ある場合は互いに同一でも異なっていてもよく、Ｒ^１は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。）
で表されるものである。 [Cobalt complex compound]
The cobalt complex compound of the present invention has the following general formula (I),

(Wherein ^A represents CO, P (R ^A ) ₃ or NO, which may be the same or different from each other, and R ^A represents the number of carbon atoms which may be substituted with a hydroxyl group, an aldehyde group, a carboxyl group or an ester group. 1-4 represents a linear or branched alkyl group; a phenyl group; an aldehyde group; a carboxyl group or an ester group, and when R ^A is 2 or more, they may be the same or different, and R ¹ is a hydrogen atom, Represents a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
It is represented by

Examples of the linear or branched alkyl group having 1 to 4 carbon atoms which may be substituted with a hydroxyl group, an aldehyde group, a carboxyl group or an ester group represented by ^RA in the general formula (I) include a methyl group, an ethyl group Group, propyl group, isopropyl group, butyl group, t-butyl group, hydroxymethyl group, formylmethyl group, carboxymethyl group, methoxycarbonylmethyl group and the like. Examples of the ester group represented by R ^A in the general formula (I) include a methoxycarbonyl group, an ethoxycarbonyl group, an n-propoxycarbonyl group, and an isopropoxycarbonyl group.

A in the general formula (I) is preferably CO or P (R ^A ) ₃ , more preferably P (R ^A ) ₃ in which CO or R ^A is a phenyl group, and particularly preferably CO.

In the general formula (I), R ¹ is preferably a methyl group.

  By reacting the cobalt complex compound with a deprotecting agent, atractyrosines represented by the general formula (II) can be easily produced.

  As described above, atractyrosines are unstable, but the cobalt complex compound of the present invention is relatively stable and hardly changes chemically. Therefore, atractyrodins can be produced by storing the cobalt complex compound of the present invention and performing a deprotection reaction when necessary.

（式中、Ｒ^２は水素原子、メチル基、ヒドロキシメチル基、アルデヒド基またはカルボキシル基を表す。）
で表されるアトラクチロジン類と、
下記一般式（ＩＩＩ）、

（式中、ＡはそれぞれＣＯ、Ｐ（Ｒ^Ａ）_３またはＮＯを表し、互いに同一でも異なっていてもよく、Ｒ^Ａは水酸基、アルデヒド基、カルボキシル基またはエステル基で置換されてもよい炭素数１〜４の直鎖または分岐鎖のアルキル基；フェニル基；アルデヒド基；カルボキシル基またはエステル基を表し、Ｒ^Ａが２以上ある場合は互いに同一でも異なっていてもよい。）
で表されるコバルト化合物とを反応させる工程を備えるものである。前記アトラクチロジン類は、合成で製造してもよく、天然物から抽出したものを用いてもよい。 [Method for producing cobalt complex compound]
The method for producing the cobalt complex compound of the present invention includes the following general formula (II),

(In the formula, R ² represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
Atractylodins represented by
The following general formula (III),

(Wherein ^A represents CO, P (R ^A ) ₃ or NO, which may be the same or different from each other, and R ^A represents the number of carbon atoms which may be substituted with a hydroxyl group, an aldehyde group, a carboxyl group or an ester group. 1-4 represents a linear or branched alkyl group; a phenyl group; an aldehyde group; a carboxyl group or an ester group, and when there are two or more R ^{A s} , they may be the same or different.
The process of making it react with the cobalt compound represented by these is provided. The atractyrosin may be produced synthetically or may be extracted from natural products.

R ² in the general formula (II) is preferably a methyl group.

The linear or branched alkyl group having 1 to 4 carbon atoms which may be substituted with a hydroxyl group, an aldehyde group, a carboxyl group or an ester group represented by ^RA in the general formula (III) is the same as described above. Things. Moreover, as an ester group which ^RA in the said general formula (III) represents, the same thing as the above is mentioned.

A in the general formula (III) is preferably CO or P (R ^A ) ₃ , more preferably P (R ^A ) ₃ in which CO or R ^A is a phenyl group, and particularly CO. preferable.

  There is no restriction | limiting in particular in the reaction temperature with the atractylazine represented by the said general formula (II), and the cobalt compound represented by the said general formula (III), It is preferable to react at room temperature-100 degreeC. The reaction time is preferably within 1 hour when the reaction temperature is room temperature, for example.

[Method for producing atractylodins]
The method for producing atractyrosin of the present invention is characterized in that the cobalt complex compound represented by the above general formula (I) is reacted with a deprotecting agent. Examples of the deprotecting agent include amine compounds such as ethylenediamine, diethylenetriamine, and triethylenetetramine, and iron (III) nitrate. Since atractyrosines can be produced with a small amount of by-products, ethylenediamine, diethylenetriamine, and triethylenetetramine. Is preferred.

  The reaction temperature with the deprotecting agent is not particularly limited, and the reaction is preferably performed at room temperature to 100 ° C. The reaction time is preferably within 1 hour when the reaction temperature is 65 ° C., for example.

  The method for producing atractylodins of the present invention preferably includes a step of sublimating and purifying the atractyrodins represented by the above general formula (II), which can be purified with a high recovery rate and a high purity. Purification can be performed by a simple operation that does not include a production control step. In addition, since no organic solvents necessary for the recrystallization method are used, in addition to the advantage that it is not necessary to pay attention to the residual solvent in the purified atractyrosin, there is also an advantage in terms of the sanitary environment of workers.

  The step of purifying by sublimation may be provided for purifying atractylodins produced by reacting the cobalt complex compound represented by the above general formula (I) with a deprotecting agent. You may provide in order to refine | purify atractyrosin used for reaction for manufacturing the cobalt complex compound represented by a formula (I).

  Sublimation is preferably performed in an environment of 0.5 hPa to 1.0 hPa, and the temperature at that time is preferably 50 to 100 ° C.

Starting materials, reagents and solvents were used without further purification unless purchased otherwise. All water-insoluble reactions were performed in an inert atmosphere (argon) with moisture removed from the reagents. The progress of the reaction was observed on a silica gel thin layer chromatography (TLC) plate, visualized under ultraviolet light, and dyed by heating with a phosphomolybdic acid solution. The product was purified by flash column chromatography (FCC) using BW-300SP (Fuji Silysia Chemical) or standard aluminum oxide 90 (Merck), or ODS column chromatography (YMC-GEL ODS-A-HG). Proton nuclear magnetic resonance spectra ( ¹ H-NMR) were recorded at 600 MHz using a spectrometer. The obtained data were evaluated for chemical shift, integral value, and multiplicity (s = single, d = doublet, dd = double doublet, t = triplet, q = quartet, br = broad, m = multiplet). Carbon nuclear magnetic resonance spectra ( ¹³ C-NMR) were recorded at 150 MHz using a spectrometer. The IR spectrum of the compound was recorded on a Jasco FT-IR 4000 using a 4000-500 cm ⁻¹ film (3M Type 61 disposable IR card). Melting | fusing point was measured using the Yanako trace amount melting | fusing point measuring apparatus.

トルエン（１．０ｍＬ）に非特許文献１に記載の合成法で得たアトラクチロジン（１７．２ｍｇ、９４．４μｍｏｌ）及びジ−コバルトオクタカルボニル（７１．０ｍｇ、２０７．７μｍｏｌ）を溶解した溶液を室温で１時間攪拌した。得られた反応混合物をシリカゲル（ヘキサン）でクロマトグラフし、黒色の固形物としてアトラクチロジンのコバルト錯体化合物を６９．０ｍｇ得た（収率９７％）。 [Experimental Example 1]
Cobalt complex compound of atractyrosin (({μ- [1 (3,4-η): 2 (3,4-η): 3 (5,6-η): 4 (5,6-η)]-( 1E, 2E) -2-nona-1,7-dien-3,5-diin-1-yl} furan) = tetrakis (tricarbonylcobalt) (Co1-Co2) (Co3-Co4))

A solution in which atractyrodine (17.2 mg, 94.4 μmol) and di-cobalt octacarbonyl (71.0 mg, 207.7 μmol) obtained by the synthesis method described in Non-Patent Document 1 were dissolved in toluene (1.0 mL). Stir at room temperature for 1 hour. The obtained reaction mixture was chromatographed on silica gel (hexane) to obtain 69.0 mg of atractylazine cobalt complex compound as a black solid (yield 97%).

The data of the obtained compound is as follows.
mp 90-92 ° C, ¹ H-NMR (CDCl ₃ , 600 MHz) δ: 1.87 (3H, s), 6.25 (1H, brs), 6.43 (2H, brs × 2), 6.62 (1H, brs), 6.83 (1H, brs), 7.20-7.33 (1H, brs), 7.45 (1H, s), ¹³ C-NMR (CDCl ₃ , 150 MHz) δ: 18.7, 92.7, 64.1, 95.7, 97.8, 109.7, 111.9, 123.9, 124.1, 127.2, 134.8, 142.9, 152.7, 198.7, IR (KBr): cm ^-1 : 2094, 2075, 2059, 2023, 2012, 2004, ESI-MS m / z: + ESI 215 [MH] +, HRESI-MS m / z: 754.7510 (Calcd for C25H11O13Co4: 754.7522).

Moreover, ¹ H-NMR of the obtained compound is shown in FIG.

ＤＭＦ（０．５ｍＬ）に上記で得たアトラクチロジンのコバルト錯体化合物（５０．１ｍｇ、６６．４μｍｏｌ）とエチレンジアミン・２Ａｃ−ＯＨ（２４２ｍｇ、１．３４ｍｍｏｌ）を溶解した溶液を６５℃で６０分間攪拌した。得られた反応混合物をシリカゲルカラムクロマトグラフィー（φ２．５＊４０、ヘキサン）で精製し、濃灰色の固形物としてアトラクチロジンを９．２ｍｇ得た（収率７６％）。 [Experimental example 2-1]
Decobaltation of cobalt complexes of atractylodyne.

A solution of the atractyrosin cobalt complex compound (50.1 mg, 66.4 μmol) and ethylenediamine · 2Ac—OH (242 mg, 1.34 mmol) obtained above in DMF (0.5 mL) was stirred at 65 ° C. for 60 minutes. did. The resulting reaction mixture was purified by silica gel column chromatography (φ2.5 * 40, hexane) to obtain 9.2 mg of atractyrosin as a dark gray solid (yield 76%).

FIG. 2 shows ¹ H-NMR of the obtained compound. It was confirmed that the ¹ H-NMR of atractyrosin obtained by decobalizing the cobalt complex compound of atractyrosin completely coincided with the ¹ H-NMR of the original atractyrosin.

[Experimental example 2-2, 2-3]
The reaction was conducted in the same manner as in Experimental Example 2-1, except that the deprotecting agent, solvent, temperature, and stirring time used were changed as shown in Table 1 below. The yield of atractylodin obtained by decobalting is shown in Table 1 below.

[Experiment 3]
Stability evaluation of cobalt complex compounds of atractyrosin 1
After the production of the cobalt complex compound of atractyrosin obtained in Experimental Example 1, a sample sealed in a light-shielding manner was stored at room temperature (July to August, no air conditioner, room temperature, humidity: going), and stability was confirmed by TLC analysis. (Thin layer plate: silica gel, developing solvent: hexane, irradiation with ultraviolet rays (254 nm)). The results are shown in FIG. It has been found that cobalt complexes of atractyrosin are stable for more than 4 months, even under relatively harsh summer room temperature and humidity.

[Experimental Example 4]
Stability evaluation of cobalt complexes of atractyrosin 2
The cobalt complex compound of atractyrosin obtained in Experimental Example 1 was subjected to the following treatment to prepare a sample.
Each sample was dissolved in hexane to prepare each sample solution. The sample solution was prepared at the time of use, and analysis was started immediately after preparation.
A. B. Store at room temperature for 100 days in a clear bottle. UV (365 nm) irradiation for 3 hours

  A calibration curve was prepared using a solution prepared by dissolving the light-blocked refrigerated product of atractylazine cobalt complex compound obtained in Experimental Example 1 in hexane. The solution used for preparing the calibration curve was prepared at the time of use, and analysis was started immediately after preparation.

The following conditions were used as HPLC analysis conditions.
<Analysis conditions>
Detection wavelength: 300 nm, column: TSK gel ODS-80Ts QA, column temperature: 20 ° C., mobile phase: acetonitrile, flow rate: 1.0 mL / min

  The calibration curve showed good linearity in the range of 0.564 μg to 1.499 μg. The quantitative values calculated using the calibration curve are shown in Table 2 below. A. Transparent bottles / stored at room temperature for 100 days; Both irradiated with ultraviolet rays (365 nm) for 3 hours gave a quantitative value of about 100%, and it was confirmed that the cobalt complex compound of atractyrosin was stable in the solid state.

[Experimental Example 5]
Sublimation purification of atractyrosin 214.0 mg of atractyrosin obtained in Experimental Example 2 was placed in 10 mL of Nascor and purified by sublimation at 80 ° C. under reduced pressure (0.73-0.81 hPa) using the apparatus shown in FIG. Went. After 1 hour, it was confirmed that the crystal was sufficiently grown, and atractyrosin adhering to the cooling tube was recovered (dry yield 202.3 mg, yield 94.5%). The conventional recrystallization purification method has a single yield rate of about 80%, and atractyrosin is obtained by concentrating the recrystallization filtrate and repeating the recrystallization. The sublimation purification method found this time has a recovery rate of over 94%. Moreover, if an apparatus is installed, it can refine | purify by a very simple operation method in a short time.

  About atractyrosin obtained by sublimation purification, the conformity of the standard for JP quantification was evaluated and confirmed to be compatible. In addition, the use equivalence with the atracty rosin reagent for quantification currently used for quality control was also confirmed.

Claims (5)

A cobalt complex compound represented by the following general formula (I):

(In the formula, A represents CO, and R ¹ represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group, or a carboxyl group.) The following general formula (II),

(In the formula, R ² represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
Atractylodins represented by
The following general formula (III),

(In the formula, A table to the CO.)
The manufacturing method of the cobalt complex compound of Claim 1 provided with the process with which the cobalt compound represented by these is made to react. The following general formula (I),

(In the formula, A represents CO, and R ¹ represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group, or a carboxyl group.)
A process for reacting a cobalt complex compound represented by the formula: and a deprotecting agent.   The method for producing atractyrosines according to claim 3, wherein the deprotecting agent is selected from the group consisting of ethylenediamine, diethylenetriamine, triethylenetetramine, and iron (III) nitrate. The following general formula (II),

(In the formula, R ² represents a hydrogen atom, a methyl group, a hydroxymethyl group, an aldehyde group or a carboxyl group.)
The method for producing atractylodins according to claim 3 or 4, comprising a step of sublimating and purifying the atractyrodins represented by the formula:

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