JPS6075294A - Production of coenzyme q - Google Patents

Abstract

PURPOSE:To provide a commercially practicable process for the production of a coenzyme Q having high purity, by treating a reduced coenzyme Q with a porous synthetic resin, and oxidizing the treated coenzyme Q. CONSTITUTION:A reduced coenzyme Q is treated with a porous synthetic resin to remove the impurities, and the treated reduced coenzyme Q is oxidized to obtain the objective coenzyme Q. A high-purity coenzyme Q10 can be produced by crystallizing a fraction of the above refined coenzyme Q having a melting point of higher than room temperature, e.g. in acetone. The treatment with the porous synthetic resin is carried out in the presence of a solvent. A polar solvent such as methanol is used for the nonpolar porous synthetic resin such as styrene divinylbenzene copolymer, etc. to effect the adsorption of a rather nonpolar impurities. On the contrary, in the case of a polar porous synthetic resin such as acrylate polymer, a nonpolar solvent such as hexane is used to effect the development and the elution.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing coenzyme Q from reduced coenzyme Q.

Coenzyme Q is a substance that is involved in the t-child transfer thread in vivo and exhibits excellent pharmacological effects against various diseases. Crude coenzyme Q can be obtained from synthesis, fermentation, or natural sources, but purifying high-purity coenzyme Q is a very unstable compound in itself, and each contains similar impurities. Therefore, it was extremely difficult. Conventionally, chromatography using inorganic materials such as silica gel, alumina, and florisil has been known as a method for purifying coenzyme Q. Chromatography using this inorganic adsorbent is a highly effective purification method for target substances containing similar impurities, but in an industrial setting, inorganic adsorbents have high specific gravity and are difficult to operate. There are many problems, and there are also problems with repeated use. Methods using polyethylene powder as an adsorbent were also known, but their surface areas were too small to be used as an industrial purification method.

The present inventors have completed the present invention as a result of various studies on commercially viable methods for producing highly pure coenzyme Q.

That is, the first invention is a method for producing coenzyme Q, in which reduced coenzyme Q, which is a raw material, is previously treated with a porous synthetic resin to remove impurities, and then the treated reduced coenzyme Q is oxidized.

Examples of porous synthetic resins used in the present invention include styrene-divinylbenzene copolymer [Amberlite XA
D-2 (manufactured by Rohm and Haas), Amberlite XAD-4 (manufactured by Rohm and Haas), High Porous Polymer HP (manufactured by Mitsubishi Chemical Corporation)]
Non-polar synthetic resin such as polyacrylic ester [Amberlite XAD-7 (manufactured by Rohm and Haas).

Amberly) XAD-8 (manufactured by Rohm and Haas)], sulfoxide [Amberlyte XAD-
9 (manufactured by Rohm and Haas)], Amide (Ami
cl, e) (Amberlight XAD-11 (ROHM)
Examples include polar synthetic resins such as A. The larger the unit surface area of these porous synthetic resins, the better, but it is usually 100 m271 or more, preferably 400 m2/2 or more. The pore size of the porous synthetic resin is preferably about 6 times or more the molecular size of the compound to be treated, usually 50λ or more, and the larger the average pore size, the better the results.

To explain the method of the present invention in more detail, the synthesis, fermentation, and extraction from natural products are reduced by conventional methods, either as they are or by adding general reducing agents such as hydrosulfide soda, sodium silane hydride, etc. After forming coenzyme Q, the porous synthetic resin is used as a carrier for chromatography with common solvents to concentrate the reduced coenzyme Q fraction in the effluent.

Next, the obtained reduced coenzyme Q is treated with a mild oxidizing agent in a conventional manner to produce coenzyme Q. Next, among the purified coenzyme Q obtained, those having a melting point above room temperature can be crystallized using a general purification method, for example, crystallized in ascent to obtain highly pure coenzyme Q10. can. In addition, examples of the reduced coenzyme Q according to the present invention include hydroquinone, which is a reduced product of coenzyme Q, as well as hydroquinone monoester, hydroquinone di-ester, etc. in which the hydroxyl group of the hydroquinone nucleus is acetylated. .

Reduced coenzyme Q is coenzyme Q mono-ester or di-ester.
In the case of an ester, it may be subjected to chromatography using a porous synthetic resin as in the case of the reduced product of coenzyme Q, followed by saponification and then subjected to an oxidation step.

Extraction and elution solvents used in the present invention include methanol, ethanol, impropatol, n-propatol, acetone, methyl ethyl ketone, impropyl ether, tetrahydrofuran, dioxane, methylcellosolve, ethyl acetate, benzene, toluene, hexane. ,
Industrially inexpensive polar or non-polar solvents such as petroleum ether, petroleum benzene, impentane, carbon tetrachloride, chloroform, dimethylformamide, water, etc. can be used alone or as a mixed solvent in various proportions. . When using a solvent, for example, for non-polar porous synthetic resins such as styrene-divinylbenzene copolymer, a polar solvent such as methanol is used to adsorb relatively non-polar impurities, and then methanol is used to adsorb relatively non-polar impurities. Add acetone to the solution and develop with a mixed solvent with weakened polarity to elute the target substance. In contrast, for polar porous synthetic resins such as acrylic acid ester polymers, they are developed and eluted using a non-polar solvent such as hexane. The resin after the elution operation can be reused by washing it with a solvent that does not adsorb any impurities. As the cleaning solvent, relatively nonpolar organic solvents such as acetone, inflovir ether, benzene, etc. are effective.

A particularly preferred method in the present invention is to use a non-polar synthetic resin as the porous synthetic resin, and use water, alcohols, ketones, dimethyl sulfoxide, etc. as the elution solvent.
A method of developing using a polar solvent such as N,N-dimethylformamide or acetonitrile is preferred. The polar solvents used in this case are generally mixed solvents based on alcohols having 1 to 5 carbon atoms and ketones having 6 to 60 carbon atoms, such as methanol and water, methanol and acetone, methanol and n-hexane, and acetone. Combinations such as carbon dioxide and water are industrially advantageous. Also, various other combinations,
Alternatively, it is of course possible to use a single solvent.

The method of the present invention is generally carried out according to the following sequence.

A column with a column length/axis ratio of 1.0 or more is filled with water, and a porous synthetic resin with an amount more than 6 times the target material (volume 7 molds) is filled by natural sedimentation. Next, the column is replaced with a solvent from which the reduced coenzyme Q is not leached out, and then the reduced coenzyme Q to be treated is allowed to flow from the top of the column. Chromatography is carried out in the same manner as in general methods, but the column may be heated if necessary if crystals of reduced coenzyme Q are precipitated. Is the effluent collected in sections and the elution category containing the target substance? The target substance can be obtained by a reduction. Next, the column is washed with a bath agent having a high elution power such as acetone, benzene, ether, esters, etc. to remove adsorbed substances, and purification chromatography can be performed again by replacing the column with a solvent.

Furthermore, unlike inorganic adsorbents, ion exchange resins, etc., the porous synthetic resin used in the method of the present invention is chemically inert. It is extremely advantageous industrially because it can be easily and completely regenerated to its original state.

The present invention has the following significantly superior effects compared to conventional methods. In other words, the separation selectivity is extremely high, and the amount of impurities adsorbed to the porous synthetic resin is extremely large, and unlike inorganic light absorbers, there is no adsorption point, so decomposition does not occur, and there is no possibility of decomposition during the processing of the porous synthetic resin. There is no loss at all, the recovery rate is extremely high, the porous synthetic resin can be used repeatedly, and unlike conventional methods that use inorganic substances and pickling agents and hydrophobic solvents such as hydrocarbons and ethers, there is no static electricity, etc. It is possible to select a safer solvent with less risk of fire due to oxidation, and highly pure coenzyme Q can be obtained by a simple purification method, which is satisfactory in terms of economic efficiency and practicality.

Next, examples of the present invention will be shown, but the present invention is not limited to the following examples.

Example 1 High porous polymer HP-20 (specific surface area 71.80
m2/t, pore volume 1.077 mg, #, 40 mesh, manufactured by Mitsubishi Chemical Industries, Ltd.) 200- was packed into a glass column (g345 + mx300), and backwashed with acetone:methanol (3N) mixture. After that, let it cool down. 2,6-Simethoxy 5- which was obtained by condensing indecaprenol and 2,3-dimethoxy-5-methyl-hydroquinone in the presence of a boron trifluoride/ether complex catalyst.
An oily substance 609 (purity 48%) containing methyl-6-zocaprenylhydroquinone (reduced coenzyme Q1o) is stirred and emulsified with 60 ml of the above-mentioned mixed solution, and the mixture is made to flow through the packed column. Then drain with the same mixture. Effluent is approximately 10
Divide into 0 m sections, perform silica gel thin layer chromatography on a very small amount of each partitioned liquid, collect the sections containing 2,6-simethoxy-5-methyl-6-decaprenylhydroquinone, and distill off the solvent by vacuum concentration. do.

The obtained treated product was dissolved in 400 mm of isopropyl ether, 20 mm of 5% potassium hydroxide was added, and air was introduced for 30 minutes at room temperature with stirring to oxidize. Next, the isopropyl ether layer is washed with water, the solvent is dried, and then distilled off under reduced pressure to obtain a red oily substance 16.1F containing coenzyme Q10. The obtained red oil was treated again with a porous synthetic resin and then subjected to thin layer chromatography to obtain coenzyme CJ1o 119t with a purity of 96%.

Example 2 High porous polymer) IP-20 (40 mesh, manufactured by Mitsubishi Chemical Industries, Ltd.) 200- was packed into a 45 fiO column and filled with acetone:methanol (2:8) mixed solution. Next, 2,3-dimethoxy-5-methyl-6-nonaprenylhydroquinone (reduced coenzyme Q9) was obtained by condensing solanesol and 2,6-simethoxy-5-methyl-hydroquinone in the presence of a zinc chloride catalyst. 302 (purity: 46%) containing oil and fat 302 (purity 46%) is added to each of the above mixed liquids 60 and stirred to emulsify, and then allowed to flow through the packed column. Then drain with the same mixture. Next, the fraction containing 2,6-simethoxy-5-methyl-6-nonaprenylhydroquinone was concentrated under reduced pressure at t7, and the treated product obtained was dissolved in 400% of inpropyl ether and 36% of lead dioxide was dissolved. and stirred for 4 hours.

After removing the lead oxide and concentrating under reduced pressure, a red oily substance 16.1' containing coenzyme Q9 is obtained.

Example 6 P5eud, 0m0naa bacteria (Pseudomonas
,s denitrificanslNRRL B-1
665 (Northern Regional Re5
[Each Laboratory, Peonria, Engineering 111onis 61604]] was collected by centrifugation, and the obtained moist soybean paste was heated and extracted with a hexane:methanol mixture in the presence of sodium hydroxide and pyrogallol. Next, the hexane layer is washed with 5-hydrosulfite soda water, further washed with water, dehydrated with sodium sulfate, and concentrated under reduced pressure. Next, the acetone-soluble part of the concentrate was taken, and the acetone was distilled off to obtain an oily substance 602 containing 2,3-dimethoxy-5-methyl-6-caprenylhydroquinone (reduced coenzyme Q). Purity 55)
Example 4 High porous polymer HP-40 (specific surface area 704.7
m2/r, pore volume 0.687 me/v, 40 mesh, manufactured by Mitsubishi Chemical Industries, Ltd.) 20 D7! was poured into a glass cylinder (045m+nX300tMIL), backwashed with an acetone:methanol (3N) mixture, and left to stand. Then add isodecaprenol and 2.3-
2,6-Simethoxy-5- produced by condensing dimethoxy-5-methyl-hydroquino/-4-monoacetate with a boron trifluoride/neethyl complex catalyst.
Oil 20f (purity 68%) containing methyl-6-zocaprenylhydroquinone-4-monoacetate (ester of reduced coenzyme QIO) is stirred and emulsified with the mixture 20- before being charged. Then drain with the same mixture. The effluent was divided into approximately 100-unit portions, and a minute amount of each fraction was subjected to silica gel thin layer chromatography to obtain 2,3-dimethoxy-5-methyl-6-zocaprenyl/
The fractions containing 1-hydroquinone-4-monoacetate are collected and the solvent is distilled off by vacuum concentration. 9.02 g of the obtained oil was dissolved in 150 g. of isopropyl ether,
Add 10-methanol containing 10% caustic potassium to this and
After standing for 0 minutes, add 30 ml of water and stir at room temperature for 60 ml.
Oxidize by introducing air for minutes. Next, the inpropyl ether layer is washed with water and the solvent is distilled off under reduced pressure to obtain 8.67 g of a red oil containing coenzyme Q10.

Patent applicant: Nisshin Seifun Co., Ltd.

Claims (1)

[Scope of Claims] 1) A method for producing coenzyme Q, which comprises treating reduced coenzyme Q with a porous synthetic resin, and then oxidizing the treated reduced coenzyme Q. 2) The method for producing coenzyme Q according to claim 1, wherein a polar solvent is used when the porous synthetic resin is a non-polar porous synthetic resin. 3) The method for producing coenzyme Q according to claim 1, wherein a non-polar solvent is used when the porous synthetic resin is a polar porous synthetic resin.