JPS61134387A - Tocol derivative - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formula I(R1-R4 are H, or lower alkyl; when R2 is CH2, R1 is not H). EXAMPLE:5, 7-Diethyl-8-methyltocol. USE:A promoting substance for normal cell division. Having improved effect and durability. PREPARATION:For example, an alpha-tocopherol shown by the formula II(R is group shown by the formula III) is reacted with chlorine under light irradiation, to give a chloromethylated derivative at the 5- and the 7-positions, which is reacted with Mg to give a compound shown by the formula IV. A carbonic acid gas is introduced into a reaction solution, which is reduced with LiAlH4, to give a hydroxyethylated derivative at the 5- and the 7-positions. The hydroxyl group at the 6-position is acetylated, and reacted with HBr, to give a compound shown by the formula V. this compound is reacted with Mg, a propyne gas is introduced to it, and the 6-position is deacetylated, to give a compound shown by the formula I(R1 is H; R2 and R3 are C2H5; R4 is CH3).

Description

Detailed Description of the Invention "Field of Industrial Application" The present invention provides novel tocol derivatives that are useful as normal cell division promoters when producing drugs such as interferon by tissue culturing animal, especially human, cells. It is related to.

"Prior Art" Conventionally, it is known that when animal, especially human, cells are subcultured in vitro, proliferation stops after about 50 generations of cell division. This is due to chromosomal mutations, etc., and this has been confirmed by testing. On the other hand, 50
There were some cells that divided and proliferated after the first generation, but in a sense, these belong to tumor cells.

However, it has been reported that when vitamin E is added to the culture medium during the above-mentioned subculture, the number of divisions of normal cells increases by about twice. Although the mechanism of action of vitamin E has not been completely elucidated, it is thought that the antioxidant effect of vitamin E, which suppresses the generation of free radicals, affects proliferation and differentiation.

Therefore, many attempts have been made to add chroman derivatives that have an antioxidant effect like vitamin E, but none of these had an effect of promoting normal cell division. Therefore, until now, only vitamin E is known to exhibit such a normal cell division promoting effect.

"Problems to be Solved by the Invention" As mentioned above, vitamin E is the only substance known to have the effect of increasing the algebraic number of one-generation cultures of normal cells. In terms of mitogenic efficacy and long-term sustainability of efficacy, it is still not fully satisfactory for practical purposes.

"Means for Solving the Problem" The present inventors focused on this point, searched for a compound other than vitamin E that has a normal cell promoting effect, and as a result of intensive research, discovered a new compound represented by the following formula. We have discovered that when added to the culture medium of humans, etc., it suppresses chromosomal mutations in the culture and prevents the culture efficiency from decreasing, and since it is less metabolized than vitamin E, its efficacy lasts for a long time.
The present invention □ has been achieved.

(In the formula, R1 to R4 represent a hydrogen atom or a lower alkyl group, but when R2 represents a methyl group, R□
does not represent a water atom) The novel compound of the present invention can be produced according to the reaction formula shown in the linear formula.
is a linear formula: Compound B can be produced by a known method of introducing chlorine into Compound A under light irradiation.

At this time, various chlorinated products such as those represented by the following formula are produced as by-products, so these can be fractionated and separated by thin layer chromatography. , Mg
is added and a known Grignard reaction is carried out, compound C
is obtained.

Compound E is obtained by introducing carbon dioxide gas into Compound C in a conventional manner to obtain Compound E, which is then reduced with LiA L Ha, HCI, and water.

Esterification of compound E by methods known per se, for example with acetic acid, gives compound F. At that time, a compound represented by the following formula is produced as a by-product, so compound F can be isolated by fractionating and branching this by thin layer chromatography. HBr is added to compound F to obtain 5-17- When the hydroxyl group of the hydroxyethyl group in position is brominated, compound G
is obtained.

Compound H is obtained by dehalogenating Compound G by a conventional method, Compound I is obtained by hydrogenating Compound H, and Compound J is obtained by hydrolyzing Compound I.

2. When R□ represents lower alkyl, (A)
(K) Compound (A)
, by reacting ethyllithium at a temperature of around 100°C,
After the reaction is completed, compound (K) is isolated by chromatographic fractionation.

In the above Reactions 1 and 2, the case where R2, R3, and R4 are methyl groups was exemplified, but it can be produced in the same manner even when these represent a hydrogen atom or a lower alkyl group.

"Examples" Next, the present invention will be further explained with reference to Examples, but the present invention is not limited to these Examples. In 0 Examples, the parts representing the quantities are parts by weight.

Example 1 100 parts of compound (A) (α-tocopherol) is dissolved in 1000 parts of chloroform, 400 parts of chlorine is introduced into the solution under ultraviolet irradiation, and the mixture is reacted at about 250° C. for 2 hours.

After completion of the reaction, the target product is isolated by chromatographic fractionation to obtain 10 parts of compound (B).

100 parts of compound (B) is dissolved in 2000 parts of anhydrous ether, and 300 parts of Mg is added thereto. Compound (C) is produced by reacting at about 100° C. for 2 hours. Two parts of carbon dioxide gas is introduced into this reaction solution at 159°C, and after one reaction is completed, 30 parts of compound (D) are separated by 11 parts by a conventional method.

100 parts of compound (D) was dissolved in 3000 parts of anhydrous ether, 4400 parts of LiA I H was added thereto, 100 fi of 35% hydrochloric acid and 3000 parts of water were added, and i
After the reaction was completed for 2 hours at SO° C., 90 parts of Compound (E) was obtained by neutralization using a conventional method.

Dissolve 100 parts of compound (E) in 3000 parts of ether,
400 parts of acetic acid was added to this, and esterification reaction was carried out at 100° C. for 4 hours. 60 parts of compound (F) was isolated from the reaction completed liquid by thin layer chromatography fractionation. Compound (F) 10 (
) in 3,000 parts of benzene, and add 35% HB to this.
Add 400 parts of r and react at 100° C. for 5 hours to produce compound CG). Next, 300 parts of Mg is added to this reaction solution, and the reaction is allowed to proceed at 100°C for 2 hours. After completion of the reaction, 90 parts of compound (H) can be obtained by isolation using a conventional method. 3,000 parts of propyne gas was introduced into the tank at 100°C, and 90 parts of compound (I) was obtained by dehalogenation and oxidation.
Standard KOH 50 parts, ether 1000 parts and water 2000 parts
Add part.

Hydrolyze at 150°C for 2 hours. Compound J (5,7-diethyl-8-
90 parts of methyl tocol were obtained. The product has an antioxidant effect, is well soluble in aliphatic solvents, and has an absorption maximum at 296 mp. b-p, 202-212°C Example 2 Compound B' in which only the methyl group at the 5-position was chlorinated was produced in the same manner as in Example 1, except that the amount of chlorine used was 1/'2. Similarly, according to Example 1, 5.7-
Diethyl-8-methyltocol was obtained as a pale yellow viscous oil. The product has an antioxidant effect, is well soluble in aliphatic solvents, and has an absorption maximum at 297°C. b,
p, 200-210°C Reference Example: To Eagle's basic medium, add 10% calf serum, 50 mg/l of aureomycin, and 100 mg/l of the additives listed in the table below, and culture human normal fibroblasts. Cultured. The culture algebra and the presence or absence of cell mutations were measured, and the results are shown in the table below. The culture number is expressed based on the maximum normal cell culture number.

As is clear from the results in the table above, the compounds of the present invention are much more effective than vitamin E in increasing the number of passages of normal cells.

"Effect" When we tested the remaining amount of additives such as vitamin E after culturing, it was tmsr/J without additives, and 10 mg/J with compound A added.
When the compound of the present invention J was added, 3 mg/ml remained. This shows that the compound of the present invention undergoes slow metabolic decomposition. This indicates that the compound of the present invention has excellent persistence of efficacy, and it is also presumed that this is the reason why it is excellent in the effect of extending the culture algebra.

"Effects of the Invention" As stated above, the novel compound of the present invention has vitamin E
Compared to other methods, it has an excellent effect on extending the culture algebra and has a long-lasting effect, so it will contribute greatly to this industry, such as by efficiently producing drugs such as interferon. From these facts, the compound of the present invention is expected to exhibit excellent effects in preventing aging.

Claims (1)

[Claims] (1) The following formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R_1 to R_4 represent a hydrogen atom or a lower alkyl group, but if R_2 represents a methyl group, R_1 represents a hydrogen atom. Tocol derivatives represented by