JPS5846506B2 - 5,6-trans-24-hydroxycholecalciferol - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing 5.6-1 lance-24-hydroxycholecalciferol.

More specifically, 5, which has calcium absorption from the small intestine.
The present invention relates to a method for producing 6-trans-24-hydroxycholecalciferol.

5,6-) Lance-24-hydroxycholecalciferol, which is the object of the present invention, is a new compound,
Its physiological effects and manufacturing method were completely unknown. ○ Active vitamin D3, an in vivo metabolic product of vitamin D3.
known as.

la, 25-dihydroxycholecalciferol or its analog theal.

la-Hydroxycholecalciferol has the effect of increasing blood calcium concentration even in nephrectomized animals (calcium absorption from the small intestine and bone mineral dissolution effect), and is associated with various disorders caused by abnormal calcium metabolism, such as renal It is known to be effective in treating bone lesions in patients with osteoporosis.

[Biochemistry
) 10.2935 (1971) and Proceedings of the National Academy of Sciences (P
roc.

Nat, Acad, Sui, ) USA, 70.
2248, (1973)).

However, all of these compounds require a hydroxyl group at the corresponding 1a-position, and require several reaction steps for 1a-hydroxylation.

Conventionally, 5゜6-dransuvitamin D3 and 5.6-trans-25-hydroxycholecalciferol are known as analogs that do not have a hydroxyl group at the 1a-position and exhibit physiological effects even in nephrectomized animals. However, like vitamin D3, 5.6-1 lance-vitamin D3 is only gradually removed from the yarn, and it seems likely that it will induce vitamin toxicity.

Further, 5,6-trans-25-hydroxycholecalciferol cannot be called an effective compound because the synthesis of 25-hydroxycholesterol, which is its raw material, is complicated.

Surprisingly, the novel 5,6-trans-24-hydroxycholecalciferol, which is the object of the present invention, rapidly promotes calcium absorption from the small intestine even in nephrectomized animals, and has the effect of increasing blood calcium concentration. The present inventors have revealed that

Thus, an object of the present invention is to provide a method for advantageously producing 5,6-)lans-24-hydroxycholecalciferol in a good yield, which is a novel compound having physiological effects useful as a pharmaceutical as described above. It is something.

According to the present invention, this object is achieved by isomerizing 24-hydroxycholecalciferol represented by the following general formula [1] by contacting it with iodine in an inert solvent, and in a high yield, the 24-hydroxycholecalciferol represented by the following general formula [1] is The following 5,6-trans-24-hydroxycholecalciferol is obtained.

The 24-hydroxycholecalciferol represented by the formula (I) used in the present invention is made from fucosterol represented by the following formula (1), which is present in large amounts in the thin film, and is made of 24-okinoculestrol, 3β, 24-diacetoxycholest-5
It can be easily produced in high yield by subjecting 3β,24-dihydroxycholester-5,7-diene obtained through -ene to ultraviolet irradiation and isomerization reaction.

This method has the advantage of being extremely advantageous compared to the production of 25-hydroxycholecalciferol, which is the raw material for 5.6-1-lans-25-hydroxycholecalciferol.

According to the present invention, the desired 5゜6-trans-2
4-hydroxycholecalciferol is obtained.

Examples of inert solvents include petroleum ether, hexane, pentane, cyclohexane, ligroin, benzene,
Toluene, xylene, chloroform, carbon tetrachloride, 1
, 2-dichloroethane, (1), 2-dibromoethane, or halogenated hydrocarbons, as well as ether solvents such as ether, tetrahydrofuran, dioxane, methylcelonorb, etc. are used.

Among these, hydrocarbon solvents such as hexane and pentane, ether solvents such as ethyl ether and tetrahydrofuran, and mixed solvents thereof are particularly preferably used.

In addition, the amount of iodine added is 24-
Against hydroxycholecalciferol, ooooi~
3% (by weight) is preferred, particularly preferably 0.001
% to 2% (by weight).

If it is less than 0.0001%, the reaction will be extremely slow;
%, other isomerization reactions are likely to occur.

As for the addition method, iodine may be added as a solid to the solvent of 24-hydroxycholecalciferol, or a solution of the splint dissolved in the above solvent may be added to the solution of 24-hydroxycholecalciferol. The addition may be reversed.

The reaction temperature is preferably in the range of -10°C to 100°C, particularly preferably in the range of 10 to 40°C.

The reaction time is usually 10 minutes to 6 hours until the reaction reaches equilibrium.

5,6-trans-24-hydroxycholecalciferol obtained by the reaction is usually treated with sodium thiosulfate (
After removing iodine with sodium thiosulfate, the crude product retained by extraction can be purified by column chromatography, preparative thin layer chromatography, high performance liquid chromatography, recrystallization, or the like.

The biological activity of 5,6-trans-24-hydroxycholecalciferol obtained by the method of the invention is as described in the following example.

Example (1) 24-hydroxycholecalciferol 16. n-hexane 1447
Dissolve in a mixed solvent of 711 and make a solution of iodine in n-hexane (
Concentration: 10 m97100 m1-n-hexane)
Added 920 μl and left the device for 3 hours 25 minutes.

After the reaction was completed, a saturated aqueous Na28203 solution was added, followed by extraction with diethyl ether.

The ether layer was washed with water, dried over Glauber's salt, and the ether was distilled off under reduced pressure to obtain 15 square meters of an oily substance.

This oily substance was separated by preparative thin layer chromatography (development; chloroform: methanol-30:
1) The result showed two bands that could be confirmed under ultraviolet light.

4.5 ■ (recovery rate 28%) of 24-
Hydroxycholecalciphenol was recovered.

5゜6-trans- exhibiting the following physical properties from the polar band
24-hydroxycholecalciphenol 8.8■ (55
%) was obtained.

U■: λ sog7' (n rn) 273 tλ so77
'(nm)232NMR(CDCA3.δ(ppm))
;0.525 (3H,S,C-18-CH3).

3.32 (IH, b, C-24-H).

3.82 (IH, b, C-3-H).

4.62 (IH, bs, C-19-H).

4.90 (IH, bs, C-19-H).

5.80 (IL(, d, JAB=11.5Hz, C
-6 or C-7H).

6.477 (IH, d, JAB-11, 5Hz, C
-6 or C-7-B).

Mass (70evm/e): 400 (M+) *2
7L255°253.136,118°High Reolution Mass:400.
3363 (M+) Theoretical value 400.3344 (C2□H4
402) (2) Regarding the 5,6-trans-24-hydroxycholecalciferol thus produced, the calcium absorption capacity from the intestinal tract and the calcium concentration in the serum were measured.

Male Wistar Weanling rats were fed a vitamin D-deficient, low-calcium diet, and after 4 weeks, both kidneys of the rats were removed and used as nephrectomized rats.

The nephrectomized rats were given 5,6-
Trans-24-hydroxycholecalciferol was administered intraperitoneally.

The control group received only an equal amount of 95% ethanol.

After 8 hours, blood was collected and the serum calcium (Ca) level was measured using a calcium measurement kit manufactured by Yatron.At the same time, the duodenum was removed and calcium was extracted from the intestinal tract using the inverted intestinal tube method according to the method of De Roux. Absorption capacity was measured [method from De Roux, American Journal Physiology (Amer, J. Physiol, )]
216.

1351 (1969)).

The results obtained are shown in Table 1.

In the table, 45ca (S/M) indicates the ratio of the calcium concentration on the intestinal mucosal side (S) to the calcium concentration on the intestinal membrane side (M), and P < 0.05 and P < 0.01 are each 5% and is significant at a 1% significance rate.

The results in this table show that 5.6-1 lance-24-hydroxycholecalciferol promotes calcium absorption from the kidneys even in nephrectomized rats, and has the effect of increasing serum calcium concentration (bone mineral dissolution effect). It shows that it has.

This indicates that this product may be effectively applied to patients with renal failure, etc.

Claims (1)

[Scope of Claims] 1 A compound represented by the following formula (II), characterized in that 24-hydroxycholecalciferol represented by the following formula (1) is brought into contact with iodine in an inert solvent to isomerize it. A method for producing 5,6-trans-24-hydroxycholecalciferol.