JPH0314009B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing polyprenyl compounds or mixtures thereof. More specifically, the present invention describes the following general formula that can be suitably used as a raw material for the synthesis of dolichol. (In the formula, A represents a hydroxyl group or an acyloxy group,

[Formula] is trans-iso represents the unit of prene,

[Formula] represents a cis isoprene unit, and n is an integer from 10 to 22. ) A method for producing a polyprenyl compound or a mixture thereof. Dolichols were introduced by JF in 1960.
It was first isolated from human kidney, pig liver, etc. by Pennock et al. [Nature (London), 186 ,
470 (1960)], and later the dolichols were defined by the following general formula: (In the formula,

[Formula] represents a trans isoprene unit,

[Formula] represents a cis isoprene unit. ), in which the number j of cis isoprene units in the above formula generally ranges from 12 to 18, with j = 14, 15
It was revealed that three homologues of 16 and 16 were the main constituents [RWKeenan et al.,
Biochemical Journal, 165 , 405 (1977)].
Furthermore, dolichols are widely distributed not only in the liver of pigs but also in the bodies of mammals, and are known to play extremely important functions in maintaining the life of living organisms. For example, JB Harford et al. conducted in vitro tests using the white medulla of calves and pigs, and found that exogenous dolichol promoted the incorporation of sugar components such as mannose into lipids, and as a result, sugars that are important for sustaining the life of living organisms It has been revealed that it has the effect of increasing protein formation [Biochemical and
Biophysical Research Communication, 76 ,
1036 (1977)]. Since the effect of dolichol on promoting the incorporation of sugar components into lipids is more pronounced in already mature animals than in living organisms during the growing stage, the role of dolichol in preventing aging is attracting attention. In addition, RWKeenan et al. state that it is important for rapidly growing organisms, such as those in infancy, to ingest dolichol from outside and supplement the dolichol obtained by biosynthesis within the body. (Archives of Biochemistry
and Biophysics, 179 , 634 (1977)]. Furthermore, Akamatsu et al. quantified dolichol phosphate ester in the regenerated rat liver, and found that the amount was significantly reduced compared to normal liver, indicating that the glycoprotein synthesis function in liver tissue was significantly reduced. We found that this function was improved by adding exogenous dolichol phosphate [presented at the 54th Annual Meeting of the Japanese Biochemical Society (1981)]. As described above, dolichols are extremely important substances for living organisms, and there is a strong desire for the development of applications in pharmaceuticals, synthetic intermediates thereof, cosmetics, and the like. However, the reality is that it has not been possible to conduct sufficient research on Dolichols because they are difficult to obtain. For example, only about 0.6 g of dolichol can be obtained from 10 kg of pig liver through complicated separation procedures [J. Burgos et al.,
See Biochemical Journal, 88 , 470 (1963)]. On the other hand, total synthesis of dolichols is difficult, as is clear from their complex and unique molecular structures.
This is extremely difficult to achieve with current organic synthetic chemistry technology. Therefore, it would be advantageous if dolichols could be obtained by relying on natural products as synthetic intermediates and adding simple synthetic chemical treatments to them, but such convenient substances have not been found so far. do not have. It has been known that polyprenol compounds can be collected from various plants, and the following polyprenols have been collected. (1) Solanesol (2) Fucaprenols (3) Betulaprenol Betulaprenol, like dolichols,
-It has a structure in which two trans isoprene units are connected to the terminal isoprene unit, but the betulaprenols known so far have only six cis isoprene units at most, as mentioned above. The number of cis isoprene units is 14
In order to synthesize dolichols, which mainly contain 1, 15 and 16 isoprene units, it is necessary to restrict and extend 8 or more isoprene units into the cis form, which is difficult to do using current organic synthesis technology. It's almost impossible. The present inventors analyzed extracts from various plants in search of plant sources for polyprenyl compounds useful as raw materials for the synthesis of mammalian dolichols.
Sciadopitydaceae, Family
Podocarpaceae), Family Podocarpaceae
A polyprenyl fraction containing a relatively high concentration of a polyprenyl compound in which A is a hydroxyl group or an acetoxy group and n is 14 to 16 in the general formula () is obtained from the leaves of plants belonging to the Cephalotaxaceae or Family Araucariaceae. that the polyprenyl fraction can be separated relatively easily into its constituent individual polyprenyl congeners, if desired, and that the polyprenyl fraction and each polyprenyl congener separated therefrom are both mammalian. It has been found that it is very suitable as an intermediate for the synthesis of dolichols. Therefore, according to the present invention, leaves of plants belonging to the family Asperaceae, Araceae, Araceae, or Araucaceae are extracted with an organic solvent, a polyprenyl compound or a mixture thereof is separated from the obtained extract, and the polyprenyl compound or a mixture thereof is separated as necessary. hydrolyzing the polyprenyl compound or mixture thereof before or after separation;
Provided is a method for producing a polyprenyl compound represented by the general formula () or a mixture thereof, which is characterized by subjecting the compound to esterification or ester conversion reaction, or two or more thereof. After extracting the leaves of plants belonging to the family Asperaceae, Araceae, Araceae, or Araucaceae with an organic solvent, and hydrolyzing the obtained extract as necessary,
Subjected to a separation method consisting of chromatography, fractional dissolution method, fractional cryoprecipitation method, molecular distillation method, or a combination of two or more of these methods,
Merck thin layer chromatography plate [silica gel
Thin layer chromatography (10 cm development) using 60F ₂₅₄ pre-coated (layer thickness 0.25 mm) and a mixed solvent of n-hexane and ethyl acetate at a volume ratio of 9:1 as the developing solvent. 0.18 to 0.25 and/or 0.50 to 0.55 under the conditions that the Rf value of solanesyl acetate as a standard substance is 0.40 to 0.45.
By isolating and collecting fractions exhibiting an Rf value within the range of and contains a substantial amount of at least three types of compounds of the general formula () where n is 14, a compound of the general formula () where n is 15, and a compound of the general formula () where n is 16 as essential components. A polyprenyl composition can be obtained. The plants used as raw materials in the present invention include plants of the family Asperaceae, plants of the family Acanthaceae, plants of the family Araceae, or plants of the family Araucaceae (hereinafter, these may simply be referred to as raw material plants). It is a plant belonging to the order Conifera. There is only one genus and one species of Sciadopitys verticillata, a plant in the family Sciadopitys verticillata, and it is mainly found only in Japan. Plants of the family Canisaceae are mainly distributed in the temperate zone and the temperate zone of the Southern Hemisphere, and are 11
It consists of several genera, especially Podocarpus
macrophylla) and Podocarpus nagi
is preferably used. Cephalotaxus Harringtonia subsp.nana is mainly distributed in East Asia and only one genus exists.
is preferably used. Araucariaceae plants exist mainly in the temperate zone and the temperate zone of the southern hemisphere, and are classified into two genera, among which Brazilian pine (Araucaria brasiliana) is preferably used. The leaves of the raw material plants may be subjected to the extraction treatment according to the present invention after drying, or they may be used undried. Dried leaves are generally preferred, and the degree of drying is advantageously such that the moisture content is generally less than about 30%, preferably less than 10%, based on the weight of the dry leaves. Furthermore, it is preferable to extract the leaves after crushing them, thereby increasing the contact area with the extraction solvent and increasing the extraction efficiency. The polyprenyl analog represented by the general formula () is generally contained in the leaves of raw material plants at a fairly high concentration in the form of free alcohol and/or acetate ester, and the polyprenyl analog is extracted from the leaves. For effective extraction, an oil-soluble organic solvent that well dissolves the polyprenyl analog is preferably used. Such oil-soluble organic solvents generally have a dielectric constant ε of 32.7 or less, preferably
25.0 or less, more preferably 20.7 or less, and specifically, the following solvents can be used alone or as a mixed solvent of two or more. (a) Hydrocarbons: for example, petroleum ether, pentane, hexane, heptane, benzene, toluene, xylene, etc. (b) Halogenated hydrocarbons: For example, chloroform, methylene chloride, carbon tetrachloride, ethane tetrachloride, perchloroethylene, trichlorethylene, etc. (c) Esters: For example, methyl acetate, ethyl acetate, ethyl propionate, etc. (d) Ethers: For example, diethyl ether, diisopropyl ether, tetrahydrofuran,
dioxane etc. (e) Ketones: For example, acetone, methyl ethyl ketone, diethyl ketone, diisopropyl ketone, etc. (f) Alcohols: e.g. methyl alcohol,
Ethyl alcohol, propyl alcohol, butyl alcohol, etc. When selecting the solvent to be used, it is desirable to use one that selectively extracts the target polyprenyl compound of the general formula () with high efficiency, and extracts as little as possible of other substances; from this point of view, Among the above solvents, hydrocarbons, halogenated hydrocarbons, esters, ethers with low polarity such as diethyl ether and diisopropyl ether, and ketones are particularly preferred. The amount of extraction solvent used is not critical and can be varied widely depending on the type of solvent used, the type and condition of the leaves to be extracted, etc., but in general, 1 part by weight of the leaves of the raw material plant (dry Approximately 1 per weight (based on weight)
It is advantageous to use within the range from about 100 parts by weight, preferably from 5 to 50 parts by weight, more preferably from 10 to 30 parts by weight. Extraction involves soaking the leaves of the raw material plant in the above solvent.
This can be carried out by stirring continuously or intermittently as necessary. The temperature during extraction is also not critical and can be varied over a wide range depending on conditions such as the type and amount of solvent used, but generally a temperature from about 0°C to the reflux temperature of the solvent can be used. and room temperature is usually sufficient. It is advantageous to carry out the extraction under such conditions, usually for 1 to 10 days. The immersion liquid after the extraction process is made into a concentrated liquid by removing the leaves and other solid content, and then removing the solvent as necessary. The extract thus obtained is then subjected to a separation process consisting of chromatography, fractional lysis, fractional cryoprecipitation, molecular distillation, or a combination of two or more of these methods to obtain the desired polyprenyl fraction. can be recovered. To confirm the fraction containing polyprenyl compounds in the above separation step, use Merck's thin layer chromatography plate (silica gel 60F ₂₅₄ pre-
thin layer chromatography (10 cm development) using a mixed solvent of n-hexane and ethyl acetate at a volume ratio of 9:1 as the developing solvent.
0.18 to 0.25 under the conditions that the Rf value of solanesyl acetate as a standard substance is 0.40 to 0.45.
[When A represents a hydroxyl group in the above general formula ()] and/or 0.50 to 0.55 [When A represents an acetoxy group in the above general formula ()]
This can be done depending on whether a spot exists at an Rf value within the range of . Therefore, in the following explanation, the Rf of thin layer chromatography
When referring to values, unless otherwise noted,
It should be understood that values measured under the above conditions are meant. The operations of chromatography, fractional dissolution, fractional cryoprecipitation, and molecular distillation methods that can be used in the extract separation process described above are known per se, and the present invention also uses known methods. Therefore, details of each method will be explained by citing literature, and only points to be particularly careful will be described here. (A) Chromatography [e.g. H. Heftman,
“Chromatography”, Reinhold Publishing Co.,
New York (1961)] Thin layer chromatography and liquid chromatography are suitable for small amounts of extracts, but column chromatography is suitable for processing large amounts of extracts. Examples of usable carriers for chromatography include silica gel, alumina, florisil, celite, activated carbon, and cellulose, of which silica gel is preferred. Examples of developing solvents when performing separation operations using a silica gel column include hexane/ethyl acetate (volume ratio 95:5 to 80:20), hexane/ethyl acetate (volume ratio 95:5 to 80:20),
Diisopropyl ether (volume ratio 95:5-80:
20), petroleum ether/methyl acetate (volume ratio 95:5
~80:20), petroleum ether/isopropyl alcohol (volume ratio 99:1~90:10), benzene/diethyl ether (volume ratio 95:5~80:20), benzene/ethyl acetate (volume ratio 98:2~ Examples include mixed solvent systems such as 80:20), chloroform, methylene chloride, etc. (B) Fractional dissolution method [e.g. LCCraig,
“Technique of Organic Chemistry”, Vol.
13, Interscience, (1951)] The polyprenyl compound of the general formula () is easily soluble in nonpolar solvents such as pentane and hexane, but poorly soluble in polar solvents such as methanol and water. Utilizing this difference in solubility, it can be purified by a fractional dissolution method. For example, impurities that are easily soluble in polar solvents are largely removed by dissolving a crude product such as an extract concentrate in the above non-polar solvent and then washing with a polar solvent that is immiscible with the non-polar solvent. can do. Examples of the nonpolar solvent preferably used in this method include hydrocarbon solvents such as petroleum ether, pentane, hexane, heptane, benzene, and toluene, and halogenated hydrocarbon solvents such as methylene chloride and chloroform. be. Further, as a polar solvent immiscible with such a non-polar solvent, water or methanol is suitable, for example. (C) Fractional cryoprecipitation method [e.g. EW Berg,
“Physical and Chemical Methods of
Separation”Chapter 14, 15, McGraw−
Hill, New York (1963)] The polyprenyl compound of the general formula () is about -
Solidifies below 10℃. Therefore, by leaving the extract under cooling at -10°C or lower, preferably about -15 to about -30°C, solidifying the target product, and then performing solid-liquid separation, it is possible to prevent it from solidifying at such temperatures. It can be purified from impurities. However, since the polyprenyl compound does not have very good crystallinity and becomes a wax-like solid, it is difficult to completely purify it by this method, so it is preferable to carry out the method in combination with other purification methods. Note that, conversely, it is also possible to preferentially precipitate and remove crystalline impurities present in polyprenol. (D) Molecular distillation method [e.g., G. Burrows,
“Molecular Distillation”, Clarendon Press,
Oxford (1960)] Since the compound of the general formula () has a large molecular weight, low molecular weight impurities can be removed by using a molecular distillation method. For example 10 ^-3 ~
Molecular distillation is carried out under a vacuum degree of 10 ^-5 mmHg and heating conditions of 100 to 250° C. to separate a low molecular fraction and a high molecular fraction. At this time, the target substance is retained in the high molecular fraction, and low molecular weight impurities can be largely removed. If a sufficiently pure polyprenyl fraction cannot be obtained by each of the above separation methods, a combination of two or more of these separation methods may be used. For example, chromatography and fractional dissolution; chromatography and fractional cryoprecipitation and fractional dissolution; chromatography and fractional cryoprecipitation, fractional dissolution and molecular distillation; chromatography and molecular distillation and fractional dissolution. Combinations of methods such as chromatography and molecular distillation; molecular distillation and fractional dissolution; molecular distillation, fractional dissolution, and fractional cryoprecipitation can be used. Thus, in thin layer chromatography
Fractions with Rf values of 0.18 to 0.25 and/or 0.50 to 0.55 are isolated and collected. The fraction with an Rf value of 0.18 to 0.25 consists essentially of a mixture of homologues of the general formula () when A represents a hydroxyl group, while the fraction with an Rf value of 0.50 to 0.55 consists of a mixture of homologs of the general formula () )
consists essentially of a mixture of homologues in which A represents an acetoxy group. Individual homolog components can also be isolated by further subjecting the thus obtained fractions to, for example, partition-type high performance liquid chromatography. In addition, in the above separation step, before subjecting the extract to the above separation operation, the extract is hydrolyzed to remove the homologs in the above general formula () where A represents an acetoxy group that may be contained therein. It is possible to convert in advance to the corresponding homologue in which A represents a hydroxyl group. This may simplify the subsequent separation operation. However, such hydrolysis occurs naturally after the separation operation is completed.
It can also be performed on fractions containing components with an Rf value of 0.50 to 0.55. This hydrolysis can be carried out using any conventional method known for hydrolyzing fatty acid esters, for example by dissolving sodium or potassium hydroxide in aqueous methanol or ethanol. solution (the alkali metal hydroxide concentration is preferably about
The above extract or fraction is added at a ratio of about 5 to 50 parts by weight to 100 parts by weight (which can be 0.1 to 30% by weight) and reacted at about 25 to 90°C for about 0.5 to 5 hours. Bye. In the polyprenyl fraction isolated and recovered by the above-mentioned method, the fraction with an Rf value of 0.18 to 0.25 is substantially composed of a mixture of multiple polyprenol homologues in which A in the general formula () represents a hydroxyl group. The fraction having an Rf value of 0.50 to 0.55 consists essentially of a mixture of a plurality of polyprenylacetate homologs when A in the general formula () represents an acetoxy group. Abundance ratio of the former and the latter in the extract (polyprenol congener mixture: polyprenylacetate congener mixture)
is approximately within the range of 20:1 to 1:100 (weight ratio), and the distribution state (distribution pattern) of polyprenol or polyprenylacetate analogs in each fraction varies depending on the type of raw material plant. The distribution of products obtained from the same species of raw material plant is generally the same, and is almost constant regardless of factors such as the youth of the leaves used as raw materials, the time of collection, and the region. When the polyprenyl composition obtained by the method of the present invention is induced into mammalian dolichols, the composition may be used as it is, or if necessary, each polyprenyl compound constituting the composition may be isolated and then reacted. You can also do that. Furthermore, the composition or the isolated polyprenyl compound, depending on whether it is in the free alcohol or acetate form, is subjected to hydrolysis, esterification or transesterification reactions or reactions of two or more thereof. Polyprenol may be converted into a polyprenyl ester, or polyprenyl acetate may be converted into another polyprenyl ester or polyprenol represented by the general formula (). As an example of a polyprenyl ester represented by the general formula () produced by the method of the present invention, when A in the general formula () is represented by RCOO-, R is a hydrogen atom, an alkyl group having 1 to 18 carbon atoms ( For example, methyl, ethyl, n-propyl, isopropyl, n
-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isoamyl, n-hexyl, n-octyl, n-decyl, n-dodecyl,
n-undecyl, stearyl, etc.), carbon number 2~
18 alkenyl groups (e.g. 3-butenyl, 3-
pentenyl, 4-pentenyl, geranyl, geranylmethyl, farnesyl, farnesylmethyl, etc.), cycloalkyl groups having 5 to 7 carbon atoms (e.g.
cyclopentyl, cyclohexyl, methylcyclohexyl, cycloheptyl, etc.), carbon number 6-10
aryl group (e.g., phenyl, tolyl, xylyl, naphthyl, etc.), aralkyl group having 7 to 11 carbon atoms (e.g., benzyl, phenethyl, methylbenzyl, dimethylbenzyl, α- or β-naphthylmethyl, etc.), or 1 to 3 Examples include esters which are methyl groups (eg, monofluoromethyl, trifluoromethyl, monochloromethyl, dichloromethyl, etc.) substituted with fluorine or chlorine atoms. The esterification or transesterification reaction of the polyprenol represented by the general formula () or a mixture thereof to obtain these polyprenyl esters or mixtures thereof can be carried out using esterification or transesterification reaction methods and conditions known per se to obtain esters. This can be done using For example, esterification is expressed by the general formula ()
Formic acid, acetic acid, monofluoroacetic acid, monochloroacetic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, caprylic acid, capric acid, in the presence or absence of a catalyst and a solvent. Lauric acid, palmitic acid, stearic acid, oleic acid,
Mix with a desired carboxylic acid such as linoleic acid, linolenic acid, farnesylic acid, farnesylacetic acid, benzoic acid, 3,5-dimethylbenzoic acid, 4-ethylbenzoic acid, or their acid halide or acid anhydride, as necessary. This can be easily done by heating and stirring. Preferably, the above polyprenol or a mixture thereof is dissolved in a solvent such as hexane, benzene, methylene chloride, chloroform, diethyl ether, etc., and 1 to 5 molar equivalents of the above acid halide relative to the polyprenol are added thereto to dissolve the polyprenol. The reaction is carried out by stirring for 1 to 24 hours at an appropriate temperature from room temperature to the reflux temperature of the solvent in the presence of 1 to 5 molar equivalents of pyridine. In addition, the transesterification reaction involves transesterification of a polyprenol represented by the general formula () or a mixture thereof with a lower alkyl ester (preferably methyl ester, ethyl ester, etc.) of a desired carboxylic acid (for example, the above various carboxylic acids). This is carried out by acting in the optional presence of a catalyst. Preferably, the polyprenol or its mixture and the desired carboxylic acid ester are dissolved in a solvent such as benzene, toluene, xylene, etc., and sodium hydride in an amount of 0.01 to 0.1 molar equivalent to the polyprenol is added for 2 hours or more. The reaction is carried out under conditions of refluxing the solvent for 5 days. The polyprenyl compound represented by the general formula () or a mixture thereof obtained by the method of the present invention can be easily converted into mammalian dolichols, for example, by the following synthetic route. In the above, X is a halogen atom, preferably Cl
or Br, and Z is a hydroxyl-protecting group such as a tetrahydropyranyl group, a methoxymethyl group,
It represents a benzyl group, etc., OCOR represents an acyloxy group, and Q represents the following group. [In the formula, n has the same meaning as that in the general formula (). ] Polyprenyl compounds and mixtures thereof obtained by the method of the present invention are useful in addition to the above-mentioned materials, for example, as cosmetic base materials, ointment base materials, or raw materials for their production. EXAMPLES The present invention will be explained below with reference to Examples, but the present invention is not limited by these Examples. In Examples and Reference Examples, IR analysis was performed using a liquid film, and NMR was measured in CDCl ₃ using TMS as an internal standard. The values of FD-MASS analysis (field ionization mass spectrometry) are ¹ H, ¹² C, ¹⁶ O,
This is a value corrected as ⁷⁹ Br. Example 1 10 kg (undried weight) of Kouyama leaves collected in mid-April in Wakayama Prefecture were dried with hot air at about 50°C for 24 hours, and then immersed in chloroform 80 at room temperature of about 15°C for extraction for one week. Hexane 5 was added to the concentrate obtained by distilling off chloroform from this extract to separate insoluble components, and the liquid was concentrated and separated using a silica gel column using chloroform as a developing solvent. =9/1
Silica gel thin layer chromatography using a mixed solvent of (volume ratio) [Merck thin layer chromatography plate (silica gel 60F ₂₅₄ coated; layer thickness 0.25
About 23 g of oil was obtained by separating the fractions containing Rf values of 0.51 and 0.20 using a 10 cm development. In addition, solanesyl acetate showed an Rf value of 0.40 in the above thin layer chromatography. Approximately 400 ml of acetone is added to this oil to dissolve the acetone-soluble components, the resulting mixture is filtered, the liquid is concentrated, and the resulting liquid is mixed with 400 ml of methanol and water.
2 hours with 40ml and 30g of sodium hydroxide65
After heating to ℃, methanol was distilled off, 400 ml of diethyl ether was added to the residue for extraction, the ether layer was washed five times with about 100 ml of saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off. hand
18.2g of liquid was obtained. Then, this liquid was separated using about 1 kg of silica gel and a mixture of n-hexane/diisopropyl ether = 90/10 (volume ratio), and was separated as a fraction showing an Rf value of 0.20 in the above thin layer chromatography. 15.3g of liquid was obtained. This liquid material was polyprenol with a purity of 90% or more, and the molecular weight distribution measured for this material was as follows. This value was determined using Merck's semi-preparative high-performance liquid chromatography column Li Chrosorb RP18-10 (C ₁₈ type), using a mixed solvent of acetone/methanol = 90/10 as the eluent, and using a differential refractometer to detect the above liquid. It is calculated from the area ratio of the chromatogram obtained by high performance liquid chromatography used as a device. Value of n Area ratio (%) (Number of cis isoprene units) 10 0.6 11 1.2 12 5.0 13 18.7 14 29.9 15 22.9 16 7.3 17 2.9 18 3.5 19 2.4 20 2.2 21 2.3 22 1.0 High performance liquid chromatography described above The Each component was fractionated from the above oily substance using a mass spectrometer, an infrared absorption spectrum, a ¹ H-NMR spectrum and ^{a 13} C-NMR spectrum. I confirmed that there is. About each ingredient
The results of FD-MASS and the δ values of ¹ H-NMR are summarized in Table 1, and the δ values of ¹³ C-NMR are summarized in Table 2. In the ¹ H-NMR data in Table 1, (b) means a broad signal, (d) a doublet signal, and (t) a triplet signal.

【table】

【table】

【table】

[Table] Example 2 After drying 5 kg (undried weight) of the leaves of A. japonica collected in late April in Okayama prefecture, they were crushed into small pieces using a mixer, and then mixed with petroleum ether/acetone = 4/1 (volume ratio) at room temperature of about 20°C. ) was extracted for 5 days using a mixed solvent of 40%. After washing the extract with water and drying it with anhydrous sodium sulfate, the solvent was distilled off to give about 100 g.
A residue was obtained. Add 1 part of n-hexane to this to dissolve the n-hexane soluble components, filter it,
After concentrating the liquid, fractions showing Rf values of 0.50 and 0.19 were collected using the same thin layer chromatography as used in Example 1 using a silica gel column with a mixed solvent of n-hexane/diethyl ether = 95/5 (volume ratio). Approximately 3.2 g of liquid material was obtained by fractionation. Approximately 50 ml of acetone is added to this liquid to dissolve the acetone-soluble components, the filtered liquid is concentrated, and the resulting liquid is heated to 65°C for 2 hours with 100 ml of methanol, 10 ml of water, and 5 g of potassium hydroxide. After that, methanol was distilled off and 200ml of hexane was added to the residue.
The hexane layer was washed five times with about 50 ml of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off to obtain 2.7 g of a liquid. Next, this liquid was separated using about 100 g of silica gel and chloroform, and the fraction showing Rf = 0.20 in the thin layer chromatography described above was obtained.
2.2g of liquid was obtained. This liquid material was polyprenol with a purity of 90% or more, and the molecular weight distribution of this material was measured using the same method as in Example 1 as shown below. Value of n Area ratio (%) (Number of cis isoprene units) 12 1.7 13 9.2 14 27.8 15 35.0 16 14.3 17 4.8 18 2.5 19 2.6 20 2.1 In addition, this liquid state was determined by the same high performance liquid chromatography as in Example 1. Each component is separated from the substance and subjected to mass spectrometry, infrared absorption spectrum, ¹ H−
When the NMR spectrum and ¹³ C-NMR spectrum were measured, it was found that the values of n in Example 1 were substantially the same as the corresponding values. Example 3 In Example 2, column chromatography was performed directly using about 100 g of silica gel and chloroform without saponification reaction using potassium hydroxide, and Rf = 0.51 was obtained in the thin layer chromatography described above. 2.1 g of liquid was separated as the fraction shown. This liquid material was polyprenylacetate with a purity of 90% or more, and the molecular weight distribution measured for this material was as follows. This molecular weight distribution was determined from the area ratio of the chromatogram obtained for the above liquid substance using the high performance liquid chromatography column used in Example 1 with acetone/methanol = 70/30 (volume ratio) as the eluent. . Value of n Area ratio (%) (Number of cis isoprene units) 12 1.5 13 9.3 14 27.9 15 35.3 16 14.4 17 4.9 18 2.3 19 2.3 20 2.1 The above liquid material (polymer Each component was separated from prenylacetate (90% or more) and analyzed by mass spectrometry, infrared absorption spectrum, ¹ H-NMR spectrum, and ¹³ C
- It was confirmed by NMR spectrum that these components were polyprenylacetate in which A is an acetoxy group in the general formula (). Table 3 shows the results of FD-MASS analysis for each component.

[Table] Furthermore, polyprenols obtained by hydrolyzing each component in the same manner as in Example 2, and polypropylenes obtained in Example 2 with the same value of n as these, Exactly the same as lenol
¹ H-NMR spectrum, ¹³ C-NMR spectrum and infrared absorption spectrum were given. Example 4 5 kg (undried weight) of Brazilian pine leaves collected in Okayama Prefecture in early April were dried with hot air and crushed into small pieces, and then crushed with ethyl acetate 40 at room temperature of about 15°C.
Extracted for days. Concentrate the extract and add approximately
500 ml of hexane was added to dissolve and remove the soluble components, and the mixture was concentrated to obtain a black liquid. This mixture was heated at 65° C. for 2 hours with 500 ml of methanol, 50 ml of water and 25 g of sodium hydroxide, then cooled to room temperature and extracted with 500 ml of hexane.
The hexane layer was washed with about 150 ml of saturated saline, dried over anhydrous magnesium sulfate, and the solvent was distilled off.
g of a liquid product was obtained. This product was purified by column chromatography using about 1 kg of silica gel and methylene chloride as a developing solution to obtain 7.2 g of a liquid product. This liquid material was polyprenol with a purity of 90% or more, and the molecular weight distribution of this material was measured using the same method as in Example 1 as shown below. Value of n Area ratio (number of cis isoprene units) 10 0.6 11 1.5 12 4.8 13 12.0 14 12.2 15 6.5 16 5.0 17 4.5 18 7.5 19 14.6 20 19.0 21 9.5 22 2.3 Also, the same high performance liquid chromatography as in Example 1 graph Each component was separated from this liquid using E, and subjected to mass spectrometry, infrared absorption spectroscopy, and ¹ H−
When the NMR spectrum was measured, it was found that the values of n in Example 1 were substantially the same as the corresponding values. Example 5 Inumaki leaves collected in Okayama prefecture in early April 5
Kg (undried weight) was subjected to the same operation as in Example 4 to obtain 5.7 g of a liquid product. This product was a polyprenol with a purity of 90% or more, and the molecular weight distribution measured by the same method as in Example 1 was as follows. Value of n Area ratio (%) (Number of cis isoprene units) 10 0.5 11 1.6 12 5.0 13 11.4 14 10.5 15 6.2 16 2.0 17 1.0 18 2.3 19 7.1 20 19.9 21 23.5 22 9.1 Also, same as in Example 1 Each component was separated from this liquid using high-performance liquid chromatography, and subjected to mass spectrometry, infrared absorption spectroscopy, and ¹ H−
When the NMR spectrum was measured, it was found that the values of n in Example 1 were substantially the same as the corresponding values. Examples 6 to 26 Leaves of Hainugaya collected in Okayama Prefecture at the end of April were dried with hot air at about 60°C for 65 hours, then divided into about 100g portions and immersed in Solvent 1 shown in Table 4 for 7 days at room temperature of about 20°C. Extracted at °C. The extraction solvent was extracted from the obtained extract, and the weight of the residue was measured and summarized in Table 4 as the total amount of extract. Dissolve these extracts in 200 ml of hexane and dilute the solution with methanol/
After washing three times with about 100 ml of a mixed solution of water = 9/1 (volume ratio), it was dried over anhydrous magnesium sulfate, and the solvent was distilled off to obtain an oil. This oil was mixed with 50 ml of methanol and 1 g of potassium hydroxide for 2 hours.65
After heating to ℃, methanol was distilled off, 100 ml of diethyl ether was added to the residue for extraction, and the ether layer was washed three times with about 50 ml of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off. An oil was obtained. Using 100g of silica gel, mix this oil with n-hexane/ethyl acetate = 9/1 (volume ratio).
A fraction (polyprenol mixture) having an Rf value of 0.19 was obtained by the same thin layer chromatography as in Example 1. The weight of this fraction is summarized in Table 4 as the polyprenol content. The composition of the polyprenol mixture obtained was substantially the same as that of the polyprenol mixture obtained in Example 2, regardless of the type of solvent used.

【table】

[Table] Example 27 In a solution in which 1.17 g of polyprenol of the general formula () where n = 14 and A = OH obtained from Koyamaki in the same manner as in Example 1 and 1.0 g of pyridine were dissolved in dry diethyl ether. Acetic anhydride at room temperature
1.2 g was added dropwise, and after the addition was completed, the mixture was stirred overnight at room temperature. The resulting reaction mixture was washed with saturated brine,
It was then dried over anhydrous magnesium sulfate and diethyl ether was distilled off to obtain a pale yellow viscous liquid. This product was purified by silica gel column chromatography (using hexane/ethyl acetate as a developing solution) to obtain 1.01 g of a pale yellow liquid. When this material was analyzed by IR, the absorption caused by the OH group of the raw material polyprenol at approximately 3300 cm ^-1 disappeared.
-Absorptions at 1745 cm ^-1 and 1255 cm ^-1 caused by OCOCH ₃ appeared. In addition, when NMR analysis was performed, the signal (doublet, δ = 4.08) attributed to -CH ₂ OH of the raw material polyprenol disappeared, and -
A new signal (doublet, δ=4.55) assigned to C H ₂ OCOCH ₃ was observed. −
The signal to be assigned to CH ₂ OCOC H ₃ is -C
Signal assigned to H ₂ −C= (δ=2.04)
It was observed to overlap with Moreover, m/e=1216 was obtained by FD-MASS analysis. From the above, this liquid has the general formula () where n = 14 and A = OCOCH ₃ .
was confirmed to be polyprenylacetate. Polyprenyl acetates with n other than 14 and polyprenyl acetate mixtures with n arbitrarily distributed between 10 and 22 could also be synthesized by similar operations. Example 28 1.17 g of polyprenol of the general formula () with n=14 and A=OH obtained from Koyamaki in the same manner as in Example 1, 0.5 g of methyl oleate and 0.01 g of sodium hydride were added to 50 ml of toluene. dissolve
It was heated at 110° C. for 24 hours under a nitrogen gas atmosphere. After the reaction solution was cooled to room temperature, it was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off to obtain a yellow liquid. This product was purified by silica gel column chromatography (using hexane/ethyl acetate as a developing solution) to obtain a 0.44%
g of a colorless viscous liquid was obtained. IR analysis of this liquid revealed that the absorption of approximately 3300 cm ^-1 due to the OH groups of the raw material polyprenol had disappeared. Also, FD−
MASS analysis gave m/e=1438. From the above, this one is n=14, A=OCOC ₁₇ H ₃₃
It was confirmed that it was a polyprenyl oleate of the general formula (). Polyprenyl oleates with n other than 14 and polyprenyl oleate mixtures with n arbitrarily distributed between 10 and 22 could also be synthesized by similar operations. Example 29 0.28 g of benzoyl chloride was added at room temperature to a mixture of 1.31 g of polyprenol of the general formula () with n = 16 and A = OH, which was obtained from Hainugaya in the same manner as in Example 2, and 10 ml of pyridine. Stir overnight at room temperature. The reaction mixture was then poured into about 150 ml of water, extracted with diethyl ether, and the resulting ether layer was washed with saturated brine, diluted hydrochloric acid, saturated sodium bicarbonate, and again saturated brine, and then poured over anhydrous magnesium sulfate. The residue was dried and diethyl ether was distilled off to obtain a yellow liquid. This was subjected to silica gel column chromatography (hexane/
(using ethyl acetate as a developing solution)
0.79g of slightly yellow liquid was obtained. IR about this stuff
As a result of analysis, the absorption caused by the OH groups of the raw material polyprenol disappeared, and absorptions caused by ester bonds appeared at 1715 cm ^-1 and 1270 cm ^-1 . Also FD
-MASS analysis gave m/e=1414.
From the above, this liquid substance has n=16 and A=
It was confirmed to be polyprenyl benzoate of the general formula (), which is OCOC ₆ H ₅ . Polyprenyl benzoates with n other than 16 and polyprenyl benzoate mixtures with n arbitrarily distributed between 12 and 20 could also be synthesized by similar operations. Reference example: In a three-necked flask purged with argon, 0.316 g (13 mmol) of magnesium strips, 0.5 ml of anhydrous tetrahydrofuran, and 1,2-dibromoethane
0.08ml was added and heated with a hair dryer until it foamed violently. Next, 2.51 g of 2-[4-bromo-3-methylbutoxy]-tetrahydro-2H-pyran
(10 mmol) of anhydrous tetrahydrofuran (3.0 ml)
The solution was added dropwise to the activated magnesium at such a rate that the solvent just boiled. After the dropwise addition was completed, the mixture was stirred at 70°C for 15 minutes. 60 ml of anhydrous tetrahydrofuran was added to this to prepare a Grignard solution. Separately, in a three-necked flask purged with argon, a solution of 6.42 g (5 mmol) of polyprenylacetate with the general formula () where n = 15 and A = -OCOCH ₃ in anhydrous tetrahydrofuran (15 ml) and a solution of Li ₂ CuCl ₄ in anhydrous tetrahydrofuran ( 0.1 molar solution, 2.0
ml) was added. The previously prepared Grignard solution was added dropwise to this at 0°C over 1 hour, and stirring was continued at 0°C for 2 hours. After that, saturated ammonium chloride water was added to this reaction mixture for hydrolysis.
Extracted with ether. The ether layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off using a rotary evaporator to obtain 7.69 g of a pale yellow liquid. This product had a main spot at Rf=0.35 by silica gel thin layer chromatography (using hexane/ethyl acetate = 97/3 as a developing solution). In addition, this pale yellow liquid substance is FD-
As a result of MASS analysis, m/e = 1284, which indicates the presence of raw material polyprenylacetate, was not detected at all, and m/e = 1284, which indicates the presence of the raw material polyprenylacetate, was not detected at all, and m/e = 1284, which indicates the compound where n = 15 and Z = tetrahydro-2H-pyranyl group in the general formula (), was not detected at all.
e=1396 was detected as the main peak. Next, dissolve this pale yellow liquid in 40 ml of hexane, and add p-toluenesulfonic acid pyridine to this.
0.13 g (0.5 mmol) and 20 ml of ethanol were added.
This solution was heated and stirred at 55°C for 3 hours. After cooling,
0.21 g of sodium carbonate was added to neutralize the mixture, and the solvent was distilled off using a rotary evaporator. The obtained concentrate was dissolved in ether, washed with saturated aqueous sodium bicarbonate, then saturated brine, and dried over anhydrous magnesium sulfate. The solvent was removed using a rotary evaporator, and the remaining oily substance was heated at 0.5 Torr. and 150° C. for 30 minutes to remove low-boiling components. The remaining liquid part was subjected to silica gel column chromatography (hexane/ethyl acetate = 9/
1 as a developing solution) to obtain 5.47 g of a colorless and transparent liquid. This product was analyzed using silica gel thin layer chromatography (hexane/ethyl acetate =
9/1 as developing solution) gave a single spot at Rf=0.19. Furthermore, the analysis results shown below confirmed that this liquid was a compound of general formula (A) where j=15. FD-MASS analysis m/e = 1312 (calculated value 1312) IR analysis (cm ^-1 ); 830, 1060, 1376, 1440,
2850, 2920, 3320 ¹³ C-NMR analysis (ppm/intensity) 135.365/430,
135.229/3567, 135.005/349, 134.937/
290, 131.210/213, 125.071/5242,
124.993/499, 124.448/505, 124.282/463,
124.214/445, 61.241/551, 40.029/541,
39.757/683, 37.548/582, 32.245/5500,
32.021/456, 29.316/528, 26.825/492,
26.699/548, 26.436/5166, 25.677/542,
25.308/567, 23.430/6330, 19.557/548,
17.679/353, 16.006/640 ¹ H-NMR analysis (ppm, signal type, proton ratio) 5.10 (b, 18H), 3.66 (m, 2H), 2.03 (b, 70H), 1.68 (s, 48H), 1.60 (s, 9H), 1.80-1.10 (m, 5H), 0.91 (d, 3H)

Claims (1)

[Scope of Claims] 1. The leaves of a plant belonging to the family Asperaceae, the family Araceae, the family Araceae, or the family Araucaceae are extracted with an organic solvent, the polyprenyl compound or a mixture thereof is separated from the resulting extract, and if necessary, before the separation. Or a general formula characterized in that the polyprenyl compound or the mixture thereof is subjected to hydrolysis, esterification, or ester conversion reaction, or two or more reactions thereof after separation. (In the formula, A represents a hydroxyl group or an acyloxy group, [Formula] represents a trans isoprene unit, [Formula] represents a cis isoprene unit, and n is an integer from 10 to 22.) A method for producing a polyprenyl compound or a mixture thereof. 2. The manufacturing method according to claim 1, wherein the organic solvent is selected from hydrocarbons, halogenated hydrocarbons, ethers, esters, and ketones. 3. Extract the leaves of plants belonging to the family Asperaceae, Araceae, Araceae, or Araucaceae with an organic solvent, hydrolyze the resulting extract as necessary, and then apply chromatography, fractional dissolution, or fractional cryoprecipitation. , using a Merck thin-layer chromatography plate (coated with silica gel 60F ₂₅₄ ; layer thickness 0.25 mm) by molecular distillation or a separation method consisting of a combination of two or more of these methods. The volume ratio of n-hexane and ethyl acetate is 9:
In the range of 0.18 to 0.25 and/or 0.50 to 0.55 under the conditions that the Rf value of solanesyl acetate as a standard substance is 0.40 to 0.45 in thin layer chromatography (10 cm development) using the mixed solvent of 1 as the developing solvent. Claim 1: A polyprenyl composition is obtained by isolating and collecting a fraction exhibiting an Rf value within
Manufacturing method described in section.