JPH0138771B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an anti-osteoporosis agent containing 24R-hydroxycholecalciferol. As the average lifespan of Japanese people increases, osteoporosis is increasing and becoming a social problem. Osteoporosis is mainly caused by an increase in bone resorption (calcium is eluted from the bones into the blood) and a decrease in calcium absorption ability in the intestinal tract, resulting in fractures of the neck of the large medullary bone, forearm fractures, compression fractures of the back vertebrae, etc. This is a disease that can easily cause bone fractures. Treatment methods include methods to improve the ability of calcium absorption in the intestinal tract, or methods to suppress bone resorption and promote bone formation. It is important to strengthen each of the latter effects. Conventional drugs used for the treatment of osteoporosis often have weak bone resorption inhibitory effects and have the risk of side effects. For example, 1α,25-dihydroxycholecalciferol (USP-3697559), 1α-hydroxycholecalciferol (USP-3741996) or
Although it is known that 1α,24-dihydroxycholecalciferol improves the ability of calcium absorption in the intestinal tract, it is not known that it has osteogenic effects, and furthermore, the dosage of these substances is high. If this occurs, there is a risk of side effects such as hypercalcemia and hypercalciuria. Furthermore, the peptide hormone calcitonin, which is used as a treatment for hypercalcemia, is known to have a bone resorption inhibitory effect, but because it is a peptide hormone, it cannot be administered orally; therefore, it cannot be administered intramuscularly. These methods are used to administer drugs, which are painful for patients. Furthermore, there are antigenicity issues when administering calcitonin of non-human origin (e.g., porcine calcitonin differs in 18 of 32 amino acids compared to human calcitonin) and the risk of anaphylaxis. It has been pointed out that there are risks such as shots. As mentioned above, no drug with a strong bone resorption inhibitory effect with few side effects has been reported to date. In view of the above points, the present inventors conducted extensive research and found that 24R-hydroxycalciferol (hereinafter referred to as the substance) suppresses the secretion of parathyroid hormone (PTH), which has a bone resorption promoting effect, for a long period of time. ,
The present invention was achieved based on the discovery that Ca absorption capacity in the intestinal tract is excellent. Osteoporosis as used in the present invention refers not only to senile and postmenopausal osteoporosis, but also to osteomalacia accompanied by osteomalacia, hyperparathyroidism, hyperthyroidism, Cushing's syndrome, diabetes, acromegaly, etc. secondary osteoporosis and rheumatoid arthritis,
Includes localized osteoporosis and bone abnormalities such as Südeck's atrophy. The substance may be administered orally or parenterally, but oral administration is preferred. Parenteral administration includes intramuscular, subcutaneous, intravenous, and rectal administration. Anti-osteoporosis drugs containing this substance as an active ingredient are available in tablets,
It is used in dosage forms such as powders, granules, suppositories, capsules, alcohol solutions, oil solutions, and aqueous suspensions. Further, as the oily solvent, corn oil, cottonseed oil, peanut oil, fish liver oil, oily ester, etc. are used. Also preferred are cacao oil and other triglycerides. Other ingredients include lactose, starch, talc,
Magnesium stearate, sugar or its derivative alcohol, physiological saline, surfactants, antioxidants, etc. may be used in combination with this substance. This substance may contain 0.000002 to 0.2 in unit dosage form.
% by weight, preferably 0.00002 to 0.002% by weight. In addition, this substance is 0.0001μg/Kg to 10μ per day.
g, preferably 0.008 μg/Kg to 1 μg/Kg. For example, per day for an adult weighing 50 to 60 kg.
The dose is 0.01 μg to 500 μg, preferably 0.5 to 50 μg. Next, we will describe the results of investigating the acute toxicity of this substance. Acute toxicity: This substance was dissolved in ethanol using 10 ddN male mice (body weight 20 ± 3 g), and the ethanol concentration was adjusted to
It was dissolved in corn oil to a concentration of 0.1% and administered orally (po). The dose is 100 μg/Kg. After observation for one week after administration, all 10 mice survived, and there was no difference in any way from the control group to which only corn oil containing 0.1% ethanol was administered. Therefore, this substance is extremely safe. Moreover, the LD ₅₀ value for oral administration is 100 μg/Kg or more. The present invention will be described in detail below based on production examples and examples. Production example This substance was prepared by the method of DeLuca et al. (Arch.Biochem.Biophys. 177 615-621 (1976))
It was synthesized using 5 gr of 24-ketocholesterol acetate () and 3 ml of ethylene glycol in 200 ml of benzene with 17 mg of p-toluenesulfonic acid in a round bottom flask equipped with a Soxhlet extractor filled with 50 gr of 4A molecular sieves. It was perfused for 1.5 hours. The reaction mixture was washed with water, dried over anhydrous magnesium sulfate, and after removing the solvent, recrystallization from acetone yielded 3.9 gr of 24-ethylenedioxycholesterol acetate (). Dissolve 3.9 gr of () in 130 ml of carbon tetrachloride, add 2.1 gr of N-bromosuccinimide,
After refluxing for 20 minutes, removing the formed precipitate and removing the solvent, the residue was dissolved in xylene, and the resulting xylene solution was added to refluxing xylene containing 3.9 ml of trimethyl phosphorite over 20 minutes. 90 more
Reflux for a minute, remove the solvent, and recrystallize with acetone.
605mg of 24-ethylenedioxycholester 5,7
-Diene-3β-ol-acetic acid ester () was obtained. 605 mg of () and 6 mg of p-toluenesulfonic acid were dissolved in 80 ml of acetone, stirred at room temperature for 5 hours, extracted with ether, and after removing the solvent, recrystallized with methanol to obtain 340 mg of 24-ketocholester-5,
7-dien-3β-ol () was obtained. 50 mg of () was dissolved in 140 ml of benzene/ethanol (2/1), irradiated with a medium pressure mercury lamp for 30 seconds, and then refluxed for 1 hour. After removing the solvent, it was impregnated with 1.5% silver nitrate. The 24-ketovitamin D ₃ acetate obtained by purification by thin layer chromatography was stirred in 3.5 ml of 5% KOH-methanol at room temperature for 1 hour, water was added, and extracted with ethyl acetate to give 8 mg.
24-ketobitane D ₃ (V) was obtained. 5 mg (V) with trace amounts of KOH and 2.3 mg NaBH ₄
Add to 2.3 ml of methanol containing
2.3 mg of NaBH ₄ was added again and stirred for an additional 30 minutes. After adding ethyl acetate, the mixture was washed three times with water, the solvent was removed, 120μ of benzene and 25μ of trimethylsilylimidazole were added, water was added after 10 minutes, the solution was extracted with ethyl acetate, and separated by thin layer chromatography. , R-form trimethylsilyl ether (TMS) was hydrolyzed in 0.4 ml of 1% KOH-methanol at room temperature for 2 hours, and purified by high performance liquid chromatography (HPLC) using 1% methanol-methylene chloride. The structural formula and properties of this substance are shown below. Molecular formula: C ₂₇ H ₄₄ O ₂ Molecular weight: 400.64 Measured value (theoretical value) Elemental analysis: C 80.90 (80.94) H 11.01 (11.07) O 8.09 (7.99) Melting point: 183-184.3℃ (183-184.5) Solvent solubility: Insoluble in water, soluble in common organic solvents such as ethanol, chloroform, acetone and fatty oils. Example 1 Calcium absorption from the intestinal tract Male Wistar rats (weight 200-300g) 5
This substance and 24R,25-dihydroxycholecalciferol were each dissolved in corn oil containing 0.1% ethanol and administered orally (po) to rats at 3 μg/Kg. On the other hand, the control group received only corn oil containing 0.1% ethanol. After a certain period of time, add radioactive calcium chloride solution ( ⁴⁵ CaCl ₂ , 30 μCi/
ml) was orally administered, and then blood radioactivity was measured over time for 10 to 60 minutes, and the maximum value was taken as an index of calcium absorption from the intestinal tract. Dosing solution volume is 0.15
ml/Kg. The dose of radioactive calcium chloride aqueous solution (PH7.0) was 0.1 ml/Kg. The results are shown in Table 1.

[Table] Example 2 PTH inhibitory effect 100 mg of this substance or 24R,25-dihydroxycholecalciferol dissolved in ethanol was administered into the carotid artery of a beagle dog weighing approximately 10 kg. peripheral blood
We investigated the temporal changes in PTH. The maximum secretion suppression of PTH was around 10% of the control level, and the time taken to return to the control level is shown in Table 2. This substance exhibits a PTH inhibitory effect and is found to have a long-lasting bone resorption inhibitory effect. Table 2 Time to return to PHH control level Main substance (24R(OH)D ₃ ) 120 minutes Comparative substance (24R, 25(OH) ₂ D ₃ ) 55 minutes Example 3 Example of anti-skeletal disorder drug 200~ 0.1g/Kg for a 300gr male Wistar rat.
5% Na-sulfacetylthiazole (SAT)
administered intraperitoneally as an aqueous solution twice a week for 4 months, while the same treatment was administered orally at 0.25 μg/Kg of 24R(OH)D ₃ daily during the last 19 days of SAT administration;
The femur was stained with titrachrome and the formation of osteoid tissue in the bone cortex was examined.
In _D3 , clear inhibition of osteoid tissue formation was observed compared to the control. As is clear from the above, this substance is useful as an anti-osteoporosis agent.

Claims (1)

[Scope of Claims] 1. An anti-osteoporosis agent containing 24R-hydroxycholecalciferol as an active ingredient. 2. The anti-osteoporosis agent according to claim 1, which is in the form of an oral dosage form. 3. The anti-osteoporosis agent according to claim 1, which is in the form of an injection.