JPS58174352A - Polyprenylnitrile - Google Patents

Abstract

NEW MATERIAL:A polyprenylnitrile shown by the formula I (group shown by the formula II is trans type isoprene unit; group shown by the formula III is cis type isoprene unit; n is 11-19). USE:A raw material for drugs, cosmetics, etc., useful as an intermediate for synthesizing especially a dolichol of mammal. PROCESS:A polyprenyl halide shown by the formula IV (X is halogen) is reacted with potassium cyanide or sodium cyanide in a polar solvent such as dimethyl sulfoxide usually at room temperature for 10-240hr, to give a compound shown by the formula I . In order to find out the progress of the reaction, the reduction in the raw material compound shown by the formula VI is advantageously and preferably traced by thin layer chromatography.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to polyprenylnitrile. More specifically, the present invention has a general formula unit of 6, where n represents an integer from 11 to 19. ) is related to a novel polyprenyl nitrile.

Polyprenyl represented by the general formula (1) provided by the present invention; tolyl is a substance useful as a raw material for medicines, cosmetics, etc., and is particularly useful as a synthetic intermediate for mammalian dolichols.

Dolichols were introduced in 1960 by J, F, P@nnoa.
It was first isolated from pig liver by Nat.
ur@(Lmdon).

[See Month IJ, 470 (1960)], which was later changed to the general formula (A) CHm CHs HjC
HH CHm -CHt-CH-CH-OH(A)C'Hs [In the formula, -CH2-C- (JI2- represents a trans isoprene sCH unit. The same applies hereinafter in this specification. ], wherein j representing the number of cis isoprene units in formula (A) is generally distributed from 12 to 181, and 3 where j = 14, 15 and 16. It was revealed that food congeners are the main component (RlWKeenan et al., B
iochemical Journal, 165.
505 (1977)]. 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, J. B. and Harford et al. conducted an in vitro test 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, the ability to sustain life in living organisms. has been shown to have the effect of increasing the formation of important glycoproteins (Biochemical and
d Biophyiical Re5earchCoI
76, 1036 (1977)
reference〕. The effect of dolichols on promoting the incorporation of sugar components into lipids is more pronounced in already mature animals than in growing organisms, so the role of dolichols in preventing aging is attracting attention.

In addition, R.W., Keenan et al. stated that it is important for rapidly growing organisms, such as those in childhood, to ingest dolichol from outside and supplement the dolichol obtained by biosynthesis within the body. [Archiveso
f Biochemistry and Biophy
sica, 179.634 (1977)] Sara K., Q. Matsura et al. quantitated 0 dolichol phosphate ester in U'to 0 Regeneration Co., Ltd. and found that the amount was significantly reduced compared to normal 0. We found that the glycoprotein synthesis function in the liver tissue was significantly reduced and that scissors function was improved when dolichol phosphate was added externally [Chapter 5].
Presented at the 4l8 Annual Meeting of the Biochemical Society (1981)].

As shown in L, dolichols are substances that control extremely important functions for living organisms, and are useful as pharmaceuticals or their intermediates. Finally 0.6
CF, W, B can only get the recall of F.
Urgos et al,, Bioch@m1cal
Journal, IIJ, 470 (1963)]. Total synthesis of dolichols is extremely difficult using current organic synthesis techniques, as is clear from their complex and unique molecular structures. It would be advantageous if dolichol II could be obtained by preserving a synthetic intermediate in a natural product and subjecting it to simple synthetic chemical treatment, but no such convenient substance has been found so far. . Conventionally, polyprenol WA (
It is known that Betula verrucola (these are called Becprenols) can be collected from Betula verrucola; It is almost impossible to synthesize dolichols using current organic synthesis techniques. 9K.

Hannus et al.
reported that a polyprenyl component was isolated from the leaves of Lveatri@) at a yield of 1 cs on a dry weight basis, and that this component was a polyprenyl acetate mixture containing 10 to 19 isoprene units primarily in the cis configuration. P
hytochemistry, 13.2563 (1
974)], their report does not elucidate the details of the trans and cis configurations in the polyprenyl acetate. Additionally, D.F.Zinkel
reported the presence of C9O polyprenols with 18 isoprene units or an average value of 18 isoprene units in the leaf extract of Pinus 5trobus (Phytochem et al.
istry, 11.3387 (1972)], this report does not provide a detailed analysis of the trans and cis configurations of wrinkled polyprenol.

Some of the inventors and their collaborators/fi, supra.

By hydrolyzing the extract extracted from ginkgo biloba and Himalayan cedar using an organic solvent, if necessary, and then treating it with salt by chromatography, fractional dissolution method, or other suitable separation method, 14 to 22 isoprene units are extracted from mammals. A polyprenyl fraction consisting of a polyprenol and/or a mixture of its acetate homologs having exactly the same trans, cis distribution as the dolichols is obtained, and the polyprenyl fraction has a saturated inbrene unit at the α-terminus compared to the mammalian dolichols. that the polyprenyl fraction exhibits a distribution of polyprenyl congeners very similar to the distribution of polyprenyl congeners in mammalian dolichols, only in the absence of We have found that scales can be relatively easily separated into polyprenyl homologues, and that both the skeleton polyprenyl fraction and each polyprenyl homologue separated therefrom are highly suitable as intermediates for the synthesis of mammalian dolichols.

In order to efficiently produce mammalian dolichols using the above polyprenyl compounds, the present inventors added f to the α-terminus of the polyprenyl chain of the polyprenyl compounds. ! As a result of intensive studies on methods for introducing inoprene units, the present invention was completed by creating polyprenyl; tolyl represented by the general formula (1), which is useful as an intermediate in such methods.

The polyprenyl nitrile of the present invention represented by the general formula (1) [hereinafter also referred to as polyprenyl nitrile (1)] has the general formula (wherein, X represents a rogen atom, and n is as defined above). polyprenyl halide (hereinafter referred to as polyprenyl halide (Il)). ] with potassium cyanide or sodium cyanide in a polar solvent.

As mentioned above, polyprenyl halide (It) is represented by the general formula (1) (wherein n is as defined above) and can be obtained directly from an extract of ginkgo or Himalayan cedar through hydrolysis. Polyprenols or mixtures thereof are combined with halogenating agents such as P
ω, phosphorus trihalide such as PBrm, 80(?#
Is it easier to halogenate with thionyl halide such as %SO&2? ! l! It will be done. This halogenation reaction is usually carried out by dissolving the above-mentioned polyprenol in a suitable solvent such as hexane or diethyl ether, and then dissolving the polyprenol in the presence or absence of a base such as triethylamine or pyridine at about -20°C. This is done by adding a halogenating agent at a temperature of +50°C.

An example of a polar solvent that can be suitably used for the reaction between polyprenyl halide (II) and potassium cyanide or sodium cyanide is dimethylsulfokind. The amount of solvent used is not critical, but it is about 3 to 100 times the weight of polyprenyl halide (It), especially 5
A range of 50 to 50 times is suitable.

Potassium cyanide or sodium cyanide is preferably used in an amount of 1 to 20 molar equivalents, particularly 2 to 10 molar equivalents, relative to polyprenyl halide (n). The desired reaction can be carried out by dissolving potassium cyanide or sodium cyanide and polyprenylno-ride (II) in the above solvent and continuing stirring.

The reaction temperature is usually room temperature, but if necessary, it can be adjusted to 0°C to 100°C.
Temperatures other than room temperature can also be employed, preferably within the range of 10°C to 50°C. The reaction time varies depending on the reaction temperature conditions employed at that time, but is usually about 10 to 240
It takes time. In order to confirm the progress of the reaction, thin layer chromatography was performed to confirm the progress of the reaction.
It is convenient and preferred to track the decrease in
Isolation of I) can be easily achieved by applying separation and purification techniques known per se. Chromatography is particularly conveniently used. Adsorbents that can be used in this chromatography include silica gel, alumina, activated carbon, and cellulose. Among these, silica gel is particularly preferably used. As the developing solvent, it is preferable to use a mixture of a small amount of a polar solvent such as diethyl ether, chloroform, ethyl acetate, or ethyl alcohol with a hydrogen sulfide solvent such as hexane, pentane, petroleum ether, or benzene.

Polyprenyl nitrile synthesized by the above method (
From 1), mammalian dolichols can be synthesized, for example, by the synthetic route shown below.

Science-CN2 Science-COzHO Science-qHtQtl Upper Science Part!
X(1) CW) (V)
(■) (■) (■) However, in the above formula, sir is the formula CH3CH3)iac H C)ia-c-CHCH2+-C82-C-C-Ckb
, S, (Cl-12-C-C-Ck-Lx% (in the formula n
is as defined above. ) represents a group represented by

Reaction (2) is a hydrolysis reaction of polyprenyl nitrile (1), and this reaction is carried out, for example, by adding 1 to 20% of polyprenyl nitrile (1) to a mixed solvent of ethylene glycol and water.
This is carried out by heating polyprenyl nitrile (1) in a solution containing twice the molar amount of sodium hydroxide or potassium hydroxide. Reaction ■ is a reduction reaction,
This reaction is the polyprenylcarboxylic acid obtained in reaction ①
rv>t, for example, by reduction with lithium aluminum hydride or the like. Reaction ■ is a conversion reaction of homoallyl alcohol to homoallyl halide,
This reaction is carried out by K when the tosylate obtained by treating the polyprenyl alcohol (■) obtained in reaction (2) with hyridine with para-toluenesulfonic acid chloride is converted with lithium bromide in acetone. Reaction ■ is C4
- This is a carburization reaction in which a Grignard reagent prepared from the polyprenyl barride <■) obtained in reaction (■) and metallic magnesium in a tetrahydro 7 run and β-methylpropiolactone are mixed in the coexistence of iodized steel. This is achieved by reacting with Polyprenyl alcohol (■), that is, mammalian dolichols, can be synthesized from K by reducing the polyprenylcarboxylic acid (■) obtained in reaction (1) with, for example, lithium aluminum hydride (reaction (2)).

Hereinafter, the present invention will be explained in more detail with reference to Examples and Reference Examples. In the Examples and Reference Examples, IR analysis was performed using a liquid film, and NMR analysis was performed using TMS as an internal standard. FD-MASS analysis values are 'H, 12C, 'N,
160. 'This is the value corrected as Br.

Reference example 1 Polyprenol content m Ginkgo biloba fil collected in Kurashiki City at the end of October.

kf (undried weight) was dried with hot air at about 40°C for 24 hours and then extracted by immersing it in chloroform Sol at room temperature (about 15°C). Petroleum ether 5j was added to the concentrate obtained by distilling off chloroform from this extract to separate insoluble components, and after concentrating the P solution, it was separated by silica gel color group using chloroform as a developing solvent to obtain an oil of about 37F. I got something. Approximately 4,001 ton of acetone was added to this oil to dissolve the acetone-soluble components, the resulting mixture was filtered, and the F solution was concentrated.
65°C for 2 hours with 0d and 20f sodium hydroxide
After heating to , methanol was distilled off, diethyl ether (500 m) was added to the residue for extraction, the ether layer was washed with about 100 d of water five times, dried over anhydrous sodium sulfate, and the solvent was distilled off. 2421 oil was obtained.

Next, this oil was mixed with n-hexane/isopropyl ether = 90/10 (volume ratio) using approximately 1 kf of silica gel.
The mixture was separated to obtain 218f oil. This oil is a polyprenol with a purity of 95 or higher,
This K is attached to a semi-area high performance liquid chromatography column LiChrosorb RP18-10 (Cts
Using a mixed solvent of acetone/methanol-90/10 (feeding ratio) as the eluent, high-performance liquid chromatography analysis was performed using a lower refractive index IT-detector. The results of calculating the area ratio of the peaks were as shown in U'F.

Peak code Number of 7s type isoprene units (e)
0.32 12
1.13 13
5.94 14
25.65 15
39.46 16
19.27 17
5.98 18
1.81 9 19 0.
8 Using this high-performance liquid chromatography, each component was separated from the above oily substance, and the components were determined to be represented by the general formula (ill) by mass spectrometry, infrared absorption spectrum, H-NMR spectrum, and llc-NMR spectrum. *iii) It is a polyprenol having a structure.

Field ionization mass spectrometry (FD-MASS) for each component
) as well as the δ value of H-NMR.
The C-NMR δ values are summarized in Table 2.

Reference Example 2 Synthesis of polyprenyl bromide Polyprenol 12.4P of general formula (@), n-15, and pyridine 1d were added to 200-n-hexane, and the resulting solution was heated at room temperature (approximately 20°C ), 2.02 g of phosphorus tribromide was added dropwise under a nitrogen gas atmosphere, and after the dropwise addition was completed, the mixture was stirred overnight at room temperature under a nitrogen gas atmosphere. Next, this n-hexane solution was put into a separating funnel, washed three times with about 50,117 g of water, dried over anhydrous magnesium sulfate, and the n-hexane was distilled off to form a slightly yellow liquid 12. I got Of.

When NMR analysis was performed on this product, a signal (
d, δ = 4.08) disappeared, and a new signal (d, δ = 3.91) assigned to PK-CHxBr appeared.

Further, this liquid material was analyzed by FD-MASS and m/e=1304. Based on these analysis results,
The above product has the general formula (IN) with nx15. X=
It was confirmed that it was polyprenyl bromide, which is Br.

By similar operations, polyprenyl bromides with n other than 15 and polyprenyl bromide mixtures with n arbitrarily distributed in the range of 11 to 19 were also synthesized.Example 1 With n=15 and X-Br Certain polyprenyl bromides of general formula (II)12. (M' was dissolved in 1001 of anhydrous dimedelsulfoxide, 0.9 f of sodium cyanide was added, and the mixture was stirred at room temperature for 3 days. The reaction solution was poured into about 5QQs4 of water, extracted with hexane, and the hexane layer was thoroughly mixed. Washed with water and then with brine.After drying with anhydrous magnesium sulfate, the solvent was distilled off using a rotary evaporator and a pale yellow liquid 11.6F
I got it. This was subjected to silica gel column chromatography [
Hexane/ethyl acetate = 98/2 (volume ratio) was used as a developing solution] to obtain a colorless liquid of 9.5 f. According to the 1F analysis results, this product has n=1 in general formula (1).
It was confirmed that the polyprenyl nitrile was polyprenyl nitrile.

IR analysis: 2250.1660.1440.1375
．． 8306R-''i-1-1-JI analysis, 88 questions
t, 53 (s, 9H).

1.62(8,45)1), 1.69(a,3H)1.
7-2.4 (m, 68H), 2.93 (d
, 2H) 5.05 (br, 18H) FD-MASS analysis: m/e=tzsx Example 2~
9 In the general formula Cu) by the same operation as in Example 1, X=
=Br and n is 11.12, 13, 14, 16.17.1
Polyprenyl nitriles having n of 11.12, 13, 14°, 16.18, and 19 in the corresponding general formula (1) were synthesized from the respective polyprenyl protons of 8 & and 19. Their yield is polyprenyl di with n=15)
It was almost the same as that when IJru was synthesized. In addition, the characteristic absorption of their infrared absorption spectra and “H-
The characteristic signal of the NMR spectrum is n at that position.
=15 substantially identical to those of the above polyprenyl nitrile. The results of FL)-MASS analysis were as follows.

2 11
9793 12
10474
13 11155
14
11836 16
13197
17 13878
18
14559 19
1523 Procedural Amendment (Voluntary) 1. Indication of the case s7 year 1ff1 permission @ @ 57864 No. 2. Name of the invention polyprenyl nitrile <+OS+ Rira Shi + I Subscription Inn Okabayashiji + J5 4. Agent phone number Bundle I: 03 (277) 31825, Subject of amendment 6 Contents of amendment (1) "At approximately 15 degrees Celsius" in the first line of page 14 of the specification
Insert "1 henmen" between "chloroform" and "chloroform", n
In the line where the value is 15, "12.37J is changed to [iS2,27J.

(3) Specification @ page 21, line 9: “1M, 18J! “It will be changed to 14, 17, and 18J.

(4) Add the following Example 10 after [1s21sJ on page 22, line 10 of the specification.

"Example 10 In the general formula (1) obtained in the same manner as Reference Example 1, n
is distributed from 11 to 19 and whose composition is substantially the same as that described in Reference Example 1 is reacted with phosphorus tribromide according to the method of Reference Example 2 to obtain a glybrenyl bromide mixture. A colorless viscous liquid was obtained by carrying out exactly the same operation as in Example 1, except that 12.0f was used in place of the mall briprenylpro[dotz,of] in the general formula (I) of Example 1.
I got 5f.

The IR1'H-NMR analysis results of this product are the results obtained for the polyprenyl nitrile of Example 1 and the characteristic absorption,
The positions of the characteristic signals were virtually identical. ”

Claims (1)

[Scope of Claims] Polyprenyl represented by the general formula (-〇H!-(!:=C-CH) represents a monopoly unit of trans isoprene, and O represents an integer from 11 to 19) Nitrile 0