JPH0230334B2 - HORIPURENIRUKAGOBUTSUOYOBISONOSEIZOHO - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to polyprenyl compounds and methods for producing the same. More specifically, the present invention relates to the following general formula () [In the formula, A represents a hydroxyl group, an acyloxy group, or a halogen atom, m represents the number of trans isoprene units, n represents the number of cis isoprene units, m is 2, and n is from 11 to 19. is an integer] and a method for producing the same. In the general formula (), when A is an acyloxy group, examples of the acyloxy group include formyloxy, acetoxy, propionyloxy, butyryloxy, hexanoyloxy, octanoyloxy, decanoyloxy, palmitoyloxy,
Preferably, it is a saturated or unsaturated fatty acid residue having 1 to 20 carbon atoms, such as stearoyloxy, oleoyloxy, and phalnesyl acetoxy.
Other acyloxy groups such as benzoyloxy groups may also be used. Further, when A is a halogen atom, the halogen atom is preferably Cl or Br. The polyprenyl compound represented by the above general formula () provided by the present invention is a new compound that has not been described in any known literature, and is useful as a raw material for the synthesis of dolichols, for example. Dolichols were first isolated from the animal body by Hemming et al. in 1960, and were later given the following general formula () (where i is the number of trans isoprene units, j
represents the number of cis isoprene units, i is 2, and j is an integer from 12 to 18), and is widely distributed in mammalian bodies. However, it is known that it plays an extremely important function in maintaining the life of living organisms. For example, JB Harford et al. have shown that exogenous dolichol promotes the incorporation of sugar components such as mannose into lipids, and as a result increases the formation of glycoproteins that are important for sustaining the life of living organisms (Biochemical and
Biophysical Research Communication 76 ,
1036 (1977)). Since the effect of promoting the incorporation of sugar components into lipids is more pronounced in already mature animals than in growing organisms, the role of dolichol in preventing aging is attracting attention. In addition, RWKeenan and colleagues state that it is important for organisms that are rapidly growing, such as during childhood, to ingest dolichol from outside and supplement the dolichol that is biosynthesized within the body. (Archives of Biochemistry and Biophysics,
179, 634 (1977)). In this way, dolichols are useful not only as pharmaceuticals, but also as biochemical test chemicals, cosmetic base materials, and the like. However, it has not been easy to obtain dolichols in the past; for example, it has to be extracted from 10kg of pig liver through a complicated separation process.
Only 0.6 g of dolichol was obtained (F. Burgos et al., Biochemical Journal, 88 ,
470 (1963)). Total synthesis of dolichols is extremely difficult with current organic synthetic chemistry techniques, as is evident from their specific molecular structures. 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. . It has been known that polyprenols 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 a terminal isoprene unit, and this is followed by a cis isoprene unit, but the betulaprenols known so far have a number of cis isoprene units as described above. There are only 6 at most, and it is almost impossible to restrict and extend 6 or more isoprene units into the cis form in order to synthesize dolichols from these. The present inventors have recently discovered that Ginkgo
biloba) from the following general formula (-a) It has been found that it is possible to collect polyprenols and/or their fatty acid esters represented by the formula (where m and n have the same meanings as in the general formula ()). Based on the above findings, extracting the polyprenol and/or its ester represented by the general formula (-a) from a plant containing the polyprenol and/or its ester using an organic solvent,
If necessary, the polyprenol and/or its ester is subjected to one or more of hydrolysis, esterification, transesterification, and halogenation to produce a polyprenyl compound represented by the general formula (). A method is provided. The novel polyprenols represented by the general formula (-a) are accumulated at high concentrations in the form of free alcohols and/or fatty acid esters in the leaves of the fig tree, and the lipid components are extracted from the leaves of the fig tree. By treating this with various separation means, it can be easily isolated in the form of free alcohol and/or fatty acid ester. Solvents that can be used to extract lipid components from Gingica leaves include hydrocarbons such as petroleum ether, petroleum benzene, pentane, hexane, heptane, benzene, toluene, xylene, methyl alcohol, ethyl alcohol, propyl alcohol, Alcohols such as butyl alcohol, chloroform, methylene chloride, carbon tetrachloride, ethane tetrachloride,
Halogenated hydrocarbons such as verchlorethylene and trichlorethylene, ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, and dioxane, ketones such as acetone, diethyl ketone, and diisopropyl ketone, methyl acetate, ethyl acetate, ethyl propionate, etc. Examples include esters of. These solvents may be used alone or in combination of two or more. Solvents other than those mentioned above can be used as long as they can dissolve the lipid components contained in the leaves of the fig tree, especially the polyprenol represented by the general formula (-a) or its fatty acid ester. Extraction is usually conveniently carried out at ambient temperature, but if desired e.g.
It is also possible to use temperatures other than ambient temperature within a wide range, such as from °C to about 200 °C. Although the extraction solvent depends on its type and composition, it is generally about 1 to 1 part by weight (dry basis) of G.
Used in a proportion of 100 parts by weight. However, it is possible to extract other usage ratios. The leaves of the fig tree to be subjected to the extraction treatment may be from young green leaves to fully autumnal leaves or leaves at any stage after falling, and these leaves may be dried or undried. By separating the extracted lipid components by chromatography, fractional dissolution, fractional cryoprecipitation, molecular distillation, gel separation, other separation methods, or a combination of two or more of these methods, the general formula The polyprenols represented by (-a) and/or their fatty acid esters are used as a mixture in which the number of cis-isoprene units, that is, the value of n, is distributed over the entire range from 11 to 19 or a part thereof, or the value of n is uniform. can be obtained as a single compound. The above polyprenols or mixtures of their fatty acid esters can be used as a mixture for the synthesis of dolichols and other uses, but if necessary, each polyprenol or its fatty acid ester can be isolated from the mixture. For example, separation using partition-type high performance liquid chromatography is effective. As mentioned above, the lipid components extracted from the leaves of the fig tree can be directly subjected to the separation process, but before the lipid components are subjected to the separation process, the fatty acid esters of polyprenol contained in the lipid components are hydrolyzed. It is preferable to convert the polyprenol into polyprenol because the separation and purification process becomes simpler. This hydrolysis can be carried out by applying any known method for hydrolyzing fatty acid esters, for example, by dissolving sodium hydroxide or potassium hydroxide in aqueous methanol or ethanol. Add the above lipids at a ratio of about 5 to 50 parts by weight to 100 parts by weight of the solution (alkali metal hydroxide concentration of about 0.1 to 30% by weight) to make about 25 to 90 parts by weight.
The reaction may be carried out at a temperature of about 0.5 to 5 hours. In addition, the general formula (-
By halogenating or esterifying the polyprenols shown in a), the corresponding polyprenyl halides or polyprenyl esters can be easily produced. In this case, known reaction methods and reaction conditions used for converting known alcohols into corresponding organic halides or esters can be applied.
For example, halogenation can be carried out in the presence or absence of an organic solvent that is inert under the reaction conditions for the polyprenols or mixtures thereof.
This can be easily carried out by adding an equivalent amount of a halogenating agent (for example, thionyl chloride, phosphorus trichloride, phosphorus tribromide, etc.) and causing the reaction. At this time, if a halogenation reaction promoter such as pyridine or triethylamine is added to the reaction system, the halogenation reaction will be further facilitated. The reaction temperature may be appropriately selected depending on the halogenating agent used within the temperature range in which it is generally used. Alternatively, the above polyprenols or mixtures thereof may be easily mixed with a large excess of concentrated hydrochloric acid or concentrated hydrobromic acid in the presence or absence of a solvent at a temperature of about 0 to 30°C. Polyprenyl halides corresponding to or mixtures thereof can be prepared. Furthermore, the corresponding polyprenyl chloride or them can be prepared by heating the above polyprenols or mixtures thereof with an equimolar or excess of a mixture of triphenylphosphine and carbon tetrachloride in the presence or absence of a solvent. It is also possible to produce mixtures of. The esterification reaction can also be carried out by esterifying the above polyprenols or mixtures thereof with formic acid, acetic acid, propionic acid, butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, palmitic acid, etc. in the presence or absence of an esterification catalyst and solvent. This can be easily carried out by mixing with an acid, a carboxylic acid such as stearic acid, oleic acid, linoleic acid, linolenic acid, pharnesyl acetic acid, benzoic acid, or an acid halide or acid anhydride thereof, and heating and stirring as necessary. It is also possible to synthesize the corresponding polyprenyl esters by using the above polyprenyl halides derived therefrom instead of the above polyprenols and reacting them with the alkali metal salts of the above carboxylic acids. . Corresponding polyprenyl esters can also be synthesized by reacting the lower alkyl esters of the carboxylic acids with the polyprenols in the optional presence of a transesterification catalyst. The polyprenyl compounds of the present invention represented by the general formula () and their mixtures can be easily led to dolichols, for example, by the following synthetic route. It has unique utility. 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., R ¹ represents an acyloxy group, and R represents the following group. Here, n has the same meaning as in general formula (). In addition to the above, the polyprenyl compounds of the present invention and mixtures thereof are useful as, for example, cosmetic base materials, ointment base materials, and raw materials for their production. EXAMPLES The present invention will be explained below with reference to Examples, but the present invention is not limited to these Examples. Example 1 10Kg of fig leaves collected in Kurashiki City at the end of October
(undried weight) was dried with hot air at about 40°C for 24 hours and then extracted by immersion in chloroform 80 at room temperature (about 15°C) for 7 days. Petroleum ether 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 to obtain about 37 g of oily material. Obtained. Approximately 400 ml of acetone is added to this oil to dissolve the acetone-soluble components, the resulting mixture is filtered, and the liquid is concentrated.
The obtained oil was heated at 65°C for 2 hours with 400 ml of methanol, 40 ml of water, and 20 g of sodium hydroxide, then the methanol was distilled off, the residue was extracted with diethyl ether (500 ml), and the ether layer was extracted with about 100 ml. After washing with water five times, it was dried over anhydrous sodium sulfate, and the solvent was distilled off to obtain 24.2 g of an oily substance. Next, this oil was mixed with n-hexane/isopropyl ether = 90/10 using about 1 kg of silica gel.
(volume ratio) of the mixed solution to obtain 21.8 g of oil. This oil was polyprenol with a purity of 95% or more, and the molecular weight distribution measured for this oil was as shown in FIG. Figure 1 shows the difference in the above oil using a semi-preparative high performance liquid chromatography column LiChrosorb RD18-10 (C ₁₈ type) manufactured by Merck & Co., Ltd. and using a mixed solvent of acetone/methanol = 90/10 (volume ratio) as the eluent. This is a chromatogram obtained by high-performance liquid chromatography using a refractometer as a detector, and each peak from 1 to 9 in the figure is m in general formula (-a).
= 2, corresponding to the values of n from 11 to 19, and their area ratios are as follows. Peak number Value of n Area ratio (%) 1 11 0.3 2 12 1.1 3 13 5.9 4 14 25.6 5 15 39.4 6 16 19.2 7 17 5.9 8 18 1.8 9 19 0.8 Using this high performance liquid chromatography, the above oily substance Each component was separated and analyzed by mass spectrometry, infrared absorption spectrum, ¹ H-NMR spectrum and ¹³ C
-NMR spectrum confirmed that these components were polyprenol having a structure represented by general formula (-a). The infrared absorption spectrum, ¹ H-NMR spectrum, and ¹³ C-NMR spectrum measured for the polyprenol with m = 2 and n = 15 in the general formula (-a) obtained here are shown in Figures 2 and 3, respectively. and shown in FIG. In addition, the results of field ionization mass spectrometry (FD-MASS) and ¹ H-NMR for each component are also presented.
The δ values of 13 C-NMR are summarized in Table 1, and the δ values of ¹³ C-NMR are summarized in Table 2. In Table 1, the m/e value of FD-MASS analysis indicates the atomic weight of carbon, hydrogen, and oxygen.
The values are corrected as ¹² C, ¹ H and ¹⁶ O. Also
In the ¹ H-NMR data, b means a broad signal, d means a doublet signal, and t means a triplet signal. [Table] [Table] Example 2 5 kg (undried weight) of Japanese yam leaves collected in Tokyo from late autumn to early winter were crushed into small pieces using a mixer, and then kept at room temperature (approximately 20°C) for 7 days as petroleum ether/acetone = 4/ 1 (volume ratio) mixed solvent (40
). After washing the extract with water and drying it with anhydrous sodium sulfate, the solvent was distilled off and the
100 g of residue was obtained. Add 1 part of n-hexane to this to dissolve the n-hexane soluble components,
After concentrating the liquid, it was roughly separated using a silica gel column with a mixed solvent of n-hexane/diethyl ether = 95/5 (volume ratio) to obtain about 17 g of an oily substance. Approximately 200 ml of acetone is added to this oil to dissolve the acetone-soluble components, and the resulting liquid is filtered to dissolve approximately 200 ml of acetone.
Separation was performed using 500 g of silica gel with a mixed solvent of n-hexane/diethyl ether = 95/5 (volume ratio) to obtain 12.1 g of an oily substance. This oily substance has a purity of 95% or more and is polyprenol acetate (hereinafter referred to as polyprenylacetate).
The molecular weight distribution measured for this product was as shown in FIG. Figure 5 shows the above oil using a high performance liquid chromatography column μBondapak/C ₁₈ manufactured by Waters Co., Ltd., and acetone/methanol =
This is a chromatogram obtained by high performance liquid chromatography using a 70/30 (volume ratio) mixed solvent as the eluent and a differential refractometer as the detector.
In the general formula (), each peak from 1 to 9 in the figure represents A=acetoxy group, m=2, and the value of n is 11
to 19 respectively, and their area ratios are as follows. Peak number Value of n Area ratio (%) 1 11 1.8 2 12 3.1 3 13 9.4 4 14 31.1 5 15 35.2 6 16 12.7 7 17 3.8 8 18 1.7 9 19 1.2 The same high performance liquid chromatography as shown in Example 1 was performed. Each component was separated from the above oil (polyprenylacetate content of 95% or more) using a method, and the components were determined by the general formula () by mass spectrometry, infrared absorption spectrum, ¹ H-NMR spectrum, and ¹³ C-NMR spectrum. The structure shown in (however, A
= acetoxy group) was confirmed to be polyprenylacetate. In the general formula () obtained here, A=acetoxy group, m=
2. The infrared absorption spectrum, ¹ H-NMR spectrum and ¹³ C-NMR spectrum measured for polyprenylacetate with n=14 are shown in FIGS. 6, 7 and 8, respectively. Also,
Field ionization mass spectrometry (FD-
MASS) results are shown in Table 3. In Table 3,
The m/e value of FD-MASS analysis is a value corrected by setting each atomic weight of carbon, hydrogen, and oxygen as ¹² C, ¹ H, and ¹⁶ O. [Table] Furthermore, polyprenols obtained by similarly hydrolyzing each component according to the hydrolysis reaction performed in Example 1 are the same as those obtained in Example 1 with the same value of n. Same as polyprenols
¹ H-NMR spectrum and infrared absorption spectrum were given. Example 3 12.4 g of polyprenol with general formula (-a), m = 2, n = 15, which was isolated from fig leaves in the same manner as in Example 1, and 1 ml of pyridine were added.
Add 2.0 g of phosphorus tribromide to the resulting solution in 200 ml of n-hexane under a nitrogen gas atmosphere at room temperature (approximately 20°C). After the dropwise addition is complete, stir overnight at room temperature under a nitrogen gas atmosphere. did. Next, this n-hexane solution was put into a separating funnel, washed 10 times with about 50 ml of water, dried over anhydrous magnesium sulfate,
A slightly yellow liquid is produced by distilling off hexane.
12.0g was obtained. When NMR analysis was performed on this material, it was found that -C H of the raw material polyprenol
₂ Signal assigned to OH group (doublet, δ=
4.08) disappeared, and a new signal (doublet, δ=3.91) assigned to -C H ₂ Br appeared.
In addition, this liquid material was analyzed using field ionization mass spectrometry (FD-
When analyzed by MASS), m/e=1304 (value corrected with each atomic weight as ¹² C, ¹ H, ¹⁶ O, and ⁷⁹ Br). According to these analysis results, the above product has the general formula () where A=Br, m=2, n=
15 was confirmed to be polyprenyl bromide. Example 4 In the general formula (-a), m=
2. When polyprenol with n=15 was reacted with thionyl chloride, 11.2 g of polyprenyl chloride with general formula () where A=Cl, m=2, and n=15 was obtained. About this thing FD−
As a result of MASS analysis, m/e=1260 ( ¹² C,
(value corrected as ¹ H, ¹⁶ O, ³⁵ Cl). Example 5 Polyprenyl bromide obtained in Example 3 0.66
g was dissolved in 10 ml of dimethylformamide, 1.0 g of anhydrous sodium acetate was added thereto, the mixture was stirred at about 50°C overnight, and then about 50 ml of diethyl ether was added and filtered. The liquid was washed 10 times with about 20 ml of water, dried over anhydrous magnesium sulfate, and the solvent was distilled off to obtain 0.58 g of a pale yellow liquid. When NMR analysis was performed on this product, the signal (doublet, δ = 3.91) attributed to -CH ₂ -Br of the raw material polyprenyl bromide disappeared, and -C
A new signal (doublet, δ=4.55) assigned to H ₂ OCOCH ₃ was observed. ―
The signal that should be assigned to CH ₂ OCOC H ₃ was not observed because it overlapped with the signal (δ = 2.04) that should be assigned to [Formula] in the polyprenyl chain. In addition, mass number m/
e=1284 (value corrected as ¹² C, ¹ H, ¹⁶ O) was obtained. According to these analysis results, the above product has the general formula () where A is an acetoxy group and m=
2. It was confirmed that it was a compound with n=15. Example 6 1.17 g of polyprenol with general formula (-a) where m = 2 and n = 14, isolated from fig leaves in the same manner as in Example 1, and 0.3 methyl oleate
g and 0.05 g of potassium hydroxide were dissolved in 50 ml of toluene and heated at 110° C. for 8 hours under a nitrogen gas atmosphere. After the reaction was completed, the reaction mixture was washed with water, dried, and then the solvent was distilled off to obtain 1.2 g of a pale yellow liquid. When this liquid substance was analyzed by FD-MASS, it was found that this substance has A in the general formula ().
is CH ₃ (CH ₂ ) ₇ CH=CH(CH ₂ ) ₇ COO group and m=
2, a mass peak m/e=1438 (value corrected as ¹² C, ¹ H, ¹⁶ O) indicating that it is a polyprenyl compound with n=14 was given. Example 7 A polyprenol with m=2 and n=14 in the general formula (-a) was produced in the same manner as in Example 6 except that 0.3 g of methyl stearate was used instead of 0.3 g of methyl oleate in Example 6. When methyl stearate and methyl stearate were transesterified,
1.2 g of a pale yellow liquid was obtained. FD−
After MASS analysis, this is the general formula ()
where A is a CH ₃ (CH ₂ ) ₁₆ COO group and m=2,
A mass peak m/e=1440 (value corrected as ¹² C, ¹ H, ¹⁶ O) was obtained, indicating that it was a polyprenyl compound with n=14.

[Brief explanation of drawings]

FIG. 1 is a chromatogram of the polyprenol mixture obtained in Example 1, obtained by partition type high performance liquid chromatography. Figure 2,
Figures 3 and 4 show the infrared absorption spectrum (Figure 2) measured for the polyprenol with 18 isobrene units isolated in Example 1, ¹ H
-NMR spectrum (Fig. 3) and ¹³ C-NMR
This is the spectrum (Figure 4). Figure 5 shows Example 2
This is a chromatogram obtained by partition-type high performance liquid chromatography of the polyprenylacetate mixture obtained in . Figures 6, 7, and 8 show the infrared absorption spectra (Figure 6) of the polyprenyl acetate with 17 isobrene units isolated in Example 2, ¹ H-
They are an NMR spectrum (Figure 7) and a ¹³ C-NMR spectrum (Figure 8).

Claims (1)

[Claims] 1. General formula [In the formula, A represents a hydroxyl group, an acyloxy group, or a halogen atom, m represents the number of trans isoprene units, n represents the number of cis isoprene units, m is 2, and n is from 11 to 19. is an integer]. 2. The polyprenyl compound according to claim 1, wherein A in the general formula is a hydroxyl group. 3. The polyprenyl compound according to claim 1, wherein A in the general formula is an acyloxy group. 4. The polyprenyl compound according to claim 3, wherein the acyloxy group is a saturated or unsaturated fatty acid residue having 1 to 20 carbon atoms. 5. The polyprenyl compound according to claim 1, wherein in the general formula, A is a halogen atom. 6. The polyprenyl compound according to claim 5, wherein the halogen atom is Cl or Br. 7 General formula [In the formula, m represents the number of trans isoprene units, n represents the number of cis isoprene units,
m is 2 and n is an integer from 11 to 19]
Extract the polyprenol and/or its ester from a plant containing the polyprenol and/or its ester using an organic solvent, and if necessary, hydrolyze the polyprenol and/or its ester. A general formula characterized by being subjected to one or more of the following treatments: A method for producing a polyprenyl compound represented by the formula [wherein A represents a hydroxyl group, an acyloxy group, or a halogen atom, and m and n have the above-mentioned meanings]. 8. The method for producing a polyprenyl compound according to claim 7, wherein the plant is a fig leaf.