JPS5810370B2 - Bifunctional terpenoids - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to the following general formula (I) [wherein n is an integer of 1 to 5, and A represents a hydroxyl group-protecting group.

This invention relates to a novel bifunctional terpenoid represented by:

The compounds of the present invention are useful as pharmaceutical intermediates.

That is, a compound obtained by removing the protective group from the compound of the present invention has an antiulcer effect.

Further, polyprenyl alcohol is a compound useful as a pharmaceutical intermediate such as a medicine such as an antihypertensive agent or a raw material for the side chain of coenzyme Q.

As shown in JP-A-53-103445, this compound can be synthesized from a hydroxy compound, which is a reduced form of the compound of the present invention, by the following carbon chain elongation reaction.

(1, m: integer) Therefore, the compound of the present invention is also useful as a raw material for the synthesis of polyprenyl alcohol.

In a chain terpenoid having a hydroxyl group (a hydroxyl group protected with a protective group) at a terminal, it is quite difficult to obtain the compound of the present invention by oxidizing the other terminal in terms of chemical synthesis.

Therefore, as a result of investigating methods using microorganisms, this was achieved by microbial oxidation using bacteria belonging to the genus Nocardia.

As a representative example of the bacteria belonging to the genus Nocaldea used in the production of the compounds of the present invention, the strain (BPM1613, Microtechnical Research Institute No. 1609) can be mentioned.

The mycological properties of this strain are as follows. The color display was based on the ``Color Standard'' published by the Japan Color Research Institute.

A Cell Morphology This strain exhibits a characteristic orange to pink color in most media, as shown in the culture characteristics below.

Young vegetative cells grow like hyphae, and branching is rarely observed.

In old cultures, hyphae breakage occurs and the bacilli are 0.4 to 0.6X.
1.3 to 2.4μ).

Positive Gram staining.

No flagella. It is not stained by acid-fast bacterial staining using the Zwill-Nielsen method.

No epiphyte of aerial mycelia was observed.

・B Properties on various media 1 Sucrose-nitrate agar medium (30°C) Growth is weak, colony color pink, no diffusible pigment.

2 Glucose-asparagine agar medium (30°C): No growth.

3 Glycerin-asparagine agar medium (30°C): weak growth, pink colony color, no diffusible pigment.

4 Starch agar medium (30°C): No growth.

5 Tyrosine agar medium (30°C): weak growth, colony color grayish white, no diffusible pigment.

6 Nutrient agar medium (30°C): Medium growth, colony color orange, no diffusible pigment.

7 Yeast: Malt agar medium (30°C): Good growth, colony color orange, no diffusible pigment.

8 Oatmil agar medium (30°C): Medium growth, colony color orange, no diffusible pigment.

9 Calcium-maleide agar medium (27°C): Medium growth, pink colony color.

10 Juicy agar slope (27℃): Medium growth, pale pink color of the village.

11 Egg-albumin slope (27℃): Growth is weak, colony color is white.

12 Potato section medium (27°C): medium growth,
The color of the village is pale orange.

13 Carrot section medium (27°C): Medium growth, pale pink color of colonies.

C Physiological properties■ Growth temperature range (nutrient agar slope): 20-42℃ 2 Liquefaction of gelatin: Negative 3 Hydrolysis of starch: Negative 4 Coagulation and peptonization of skim milk: Negative 5 Lidmus milk: No change 6 Melanin-like pigment Production: Negative 7 Reduction of nitrate: Positive 8 L-arabinose, D-xylose, D-glucose, D-fructose, sucrose, D-mannite, glycerin, lactose, D-galactose, D-
No acid or gas production from mannose, maltose, trehalose or Tenman.

9 Catalase test: Negative 10 Indole production: Negative 11 Hydrogen sulfide production: Negative D Assimilation of each carbon source (on Pridham-Gotliep agar, 30°C, 7 days) L-arabinose (a), D-xylose ( -+), D
-glucose (t), D-fructose (t), sucrose, inosyto-/L-(+), L-rhamnose (-), raffinose (-), D-mannitol ((1)
, +: Medium growth, +: Slight growth, -: No growth.
ral Applied Microbiology,
9.119 (1963): The Actino
mycetales, TheJena Inter
national symposium on taxo
Glycerol Kelner Molton (Glycerol Kelner Molton)
When we measured the infrared absorption spectrum of the bacterial cells cultured in Morton) medium, we found absorption bands characteristic of the Nocaldea genus (I: C, E type, ■: C type, ■: C type, ■: D
type) was given.

The above properties are described in Berey's Manual of Ddterm.
Inative Bacteriology) 7th edition or The Actinomycete by Waxman
When examined using Volume 2 of S., it was found that it belongs to the genus Nocaldea.

Next, a method for producing the compound of the present invention will be described.

It belongs to the genus Nocaldea and has the general formula (n) [where n is an integer of 1 to 5, and A represents a hydroxyl protecting group.

] A bacterium having the ability to oxidize a compound represented by the general formula (I[
) is cultured in a medium containing the compound of formula (I) as the main carbon source, and the cultured product is synthesized by the general formula (I) [where n and A have the above meanings.

The compound of the present invention represented by ] is collected.

As A in the general formulas (I) and (II), a usual protecting group for a hydroxyl group can be used.

Examples include benzyl group, tetrahydropyranyl group, methoxymethyl group, and methoxyethoxymethyl group.

As the bacteria belonging to the genus Nocaldea used in the present invention, any bacteria can be used as long as it has the ability to oxidize the compound of general formula (II).

An example of such a bacterium is the above-mentioned strain (BPM1613, Microtechnical Research Institute No. 1609).

Specifically speaking about the culture method, the culture source is:
Other than the compound of general formula (n) which is the main carbon source, commonly used compounds can be used.

Examples of nitrogen sources include nitrates such as potassium nitrate, sodium nitrate, and ammonium nitrate, ammonium salts such as ammonium chloride and ammonium phosphate, ammonia, and urea.

In addition, as necessary, inorganic salts such as potassium phosphate, sodium phosphate, magnesium sulfate, iron sulfate, manganese sulfate, etc. may be added, and organic nutritional sources such as vitamins, amino acids, or yeast extract containing these, corn steep liquor, and wort may be added. etc. are added.

The pH of the medium is usually good on the alkaline side of 7 to 10.

The culture temperature is 20 to 40°C, and the culture can be carried out under aerobic conditions such as aerated agitation culture for 3 to 5 days.

After completion of the culture, the compound of the present invention can be collected by extracting the culture with an organic solvent.

Ethyl ether, benzene, chloroform, etc. are used as the extraction solvent.

The compound of the present invention can be separated and purified by silica gel column chromatography.

Unreacted raw materials can be recovered by the above extraction operation and column chromatography (recovery rate of about 80-90%).
, which can be used as raw material again.

Next, the present invention will be explained in more detail by showing Examples and Experimental Examples.

Experimental example 1 Production of raw material compound (1) Tetrahydropyranyl ether 3.7.11.15-tetramethyl-2.6.10.1
15 g of 4-hexadecatetraen-1-ol and 1.5 g of para-toluenesulfonic acid were dissolved in methylene chloride, and 8.7 g of 2,3-dihydropyran was added dropwise over 30 minutes at 0 to 5°C while stirring. did.

After stirring at 0 to 5° C. for 30 minutes, the mixture was transferred to a separating funnel and washed with an aqueous sodium carbonate solution.

The solution was concentrated, and the concentrate was purified by silica gel column chromatography to obtain ■-(2-tetrahydropyranyl)oxy-3,7,11,15-tetramethyl-2.
13 g of 6,10,14-hexadecatetraene (yield: 6
8%).

Other tetrahydropyranyl ethers were also obtained by the same method as above.

(2) Benzyl ether 3,7,11,15-tetramethyl-2,6,10,1
4-Hexadecatetraen-1-ol: 6.0 g of potassium hydroxide, which had been reduced to 15 g, was added to 100 ml of benzyl chloride, and the mixture was refluxed for 2 hours with stirring.

After cooling, hexane 11 was added, washed with water, and concentrated.

The concentrate was purified by silica gel column chromatography to obtain 12t of 1-benzyloxy-3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraene (yield: 61%). Ta.

Other benzyl ethers were also obtained in the same manner as above.

Compound of Example 1 1%, NH4N030.25%, KH
2PO40.15%, Na2HPO40.15%, Mg
SO4・7H200,05%, FeSO4・7H20o
, ooi%, CaCl2.2H200,001%, and yeast extract 0.002%.
H7,2), belonging to the genus Nocaldea (BPMI
613, Microtechnical Research Institute No. 1609) Seed culture solution of 8 strains.
% (volume), and cultured with shaking at 30° C. for 5 days in a 500 ml flask with a shoulder.

After the cultivation was completed, the culture was extracted with ethyl ether under acidic sulfuric acid (pH 2).

The solvent was distilled off, and the residue was purified by column chromatography on silica gel.

Hexane and ethyl ether were used as developing solvents.

The obtained compounds are shown in Table 2 below.

Note that NMR was measured using some of the compounds as methyl esters.

Experimental Example 2 Production of final compound (removal of protecting group) 6 ml of hiricine was added to 2.35 f of tetrahydropyranyl ether p-toluenesulfonic acid, and the mixture was stirred at room temperature for 20 minutes.

After evaporating the solvent and washing the residue with acetone, it was dissolved in water (
3.15 mg/ml).

2O-(2-tetrahydropyranyl) obtained in the above (a)
Oxy-2, 6, 10, 14, 18-pentamethyl-2
・Add 4 rul of the above aqueous solution to 100 mg of 6, 10, 14, 18-eicosapentaenoic acid, and heat at 55°C.
The mixture was stirred for 3 hours.

After cooling, extraction was performed with 50 ml of nityl ether, and the solvent was distilled off to give 20-hydroxy-2, 6, 10, 14, 18
-Pentamethyl-2,6,10,14,18-eicosapentaenoic acid 82~ was obtained.

The protecting groups of other tetrahydropyranyl ethers obtained in the precursor a) were removed in the same manner as in this method.

■Metal lithium wire (containing 1% sodium) 1.
7'iI was cut into small pieces and added in small portions.

After raising the temperature to -20°C to completely dissolve the lithium, it was cooled again to -78°C.

16-benzyloxy-2,6,10 obtained in the above (a)
- 5v of 14-tetramethyl-2,6,10,14-hexadelic acid was dissolved in 5oml of tetrahydrone, and this was added dropwise to the above lithium solution over 30 minutes.

After stirring for 30 minutes, 1,3-butadiene was bubbled into the solution until the color (blue) of the solution faded.

Methanol was added to the resulting yellow solution until the color faded, and then the temperature was gradually warmed to room temperature.

A lot of the generated solid was collected and dissolved in water.

Under water cooling, IN hydrochloric acid was added to the aqueous solution to make it weakly acidic, and then extracted with hexane.

The extract was washed successively with water and saturated brine, and then dried by adding magnesium sulfate.

The solution was concentrated, and the concentrate was purified by column chromatography on silica gel to obtain 16-hydroxy-2.6.10
- 3.3 g of 14-tetramethyl-2,6,10,14-hexadecatetraenoic acid was obtained.

The protecting groups of other benzyl ethers obtained in the above αa) were removed in the same manner as in this method.

The obtained compounds are shown in Table 3 below.

In the table, the * mark in the yield section indicates the yield of the protective film removal reaction of the benzyl ether in (1) above, and the unmarked mark indicates the yield of the protective group removal reaction of the tetrahydropyranyl ether in (2) above.

The final compound obtained above has anti-ulcer activity as shown below.

Effect on cold restraint stress ulcer SD rats (female, body weight approximately 1701.8 to 10 weeks old) were used as test animals, and Levine's method [Proc, S
oc, Exptl, BioloMed, Vol. 124, p. 1221 (1967)], the inhibitory effect of the test compound on the development of cold restraint stress ulcers was measured.

The following were selected as test compounds. 12-hydroxy-2,6,10-)limethyl-2,6,10-dodecatrienoic acid (compound A) 16-hydroxy-2,6,10,14-tetramethyl-2,6,10,14 -Hexadelic tetraenoic acid (compound B) 20-hydroxy-2,6,10,14,18-pentamethyl-2,6,10,14,18-eicosapentaenoic acid (compound C) The table shows the inhibition rate of cold restraint stress ulcer occurrence.

Claims (1)

[Scope of Claims] 1 General Formula (I) [In the formula, n is an integer of 1 to 5, and A represents a hydroxyl group-protecting group. ] Difunctional terpenoids. 2. The compound according to claim 1, wherein A is a benzyl group or a 2-tetrahydropyranyl group. 38-(2-tetrahydropyranyl)oxy-2,6-
3. The compound according to claim 2, which is dimethyl-2,6-octazininoacide. 412-(2-tetrahydropyranyl)oxy-2.6
- The compound according to claim 2, which is 10-trimethyl-2,6,10-dodecatrienoic acid. 512-Benzyloxo-2,6,10-trimethyl-
The compound according to claim 2, which is 2,6-10-dodecatrienoic acid. $16-(2-tetrahydropyranyl)oxy-2.6
- The compound according to claim 2, which is 10,14-tetramethyl-2,6,10,14-hexadenate tetraenoic acid. The compound according to claim 2, which is 716-benzyloxy-2,6,10,14-tetramethyl-2,6,10,14-hexadenate tetraenoic acid. 82O-(2-tetrahydropyranyl)oxy-2.6
・10.14.18-pentamethyl-2.6.10.1
The compound according to claim 2, which is 4,18-eicosapentaenoic acid. 920-benzyloxy-2, 6, 10, 14, 18-
The compound according to claim 2, which is pentamethyl-2,6,10,14,18-eicosapentaenoic acid. 1024-(2-tetrahydropyranyl)oxy-2.
6.10.14.18.22-hexamethyl-2.6.
The compound according to claim 2, which is 10,14,18,22-tetracosahexaenoic acid.