JPS6221331B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an antihypercalcemic agent containing 24-hydroxycholecalciferol. Calcitonin has conventionally been used as a therapeutic agent for hypercalcemia. Since calcitonin is a peptide hormone, it cannot be administered orally, and is administered by intramuscular injection, but this method has the drawback of causing pain to the patient. Furthermore, since it has a short internal lifespan, its effects are less durable, and it has been pointed out that administering calcitonin of non-human origin poses risks such as reduced efficacy due to antibody production and anaphylactic shock. In view of the above points, the present inventors conducted extensive studies and found that 24-hydroxycholecalciferol (hereinafter referred to as the present substance or 24-(OH) _-D3 ) is orally administrable and has excellent long-lasting effects. The present invention was achieved by discovering that The effect of this substance on serum calcium is as follows:
Already, Ikegawa et al. (Biochemistry, 14 , (15),
3293-3296, 1975). This report concerns rats raised on a low-calcium, vitamin D-deficient diet, and states that administration of this substance increases serum calcium. That is, it is known that this substance has the effect of increasing the serum calcium concentration in patients suffering from hypocalcemia symptoms. However, this substance lowers serum calcium concentration even in hypercalcemic states,
It is not yet known that it has the effect of restoring normal conditions. The present inventors have discovered that this substance restores abnormal serum calcium levels to normal levels,
Moreover, it was found that it lasted for a long time. Therefore, this substance can be said to be useful as an antihypercalcemic agent. Hypercalcemia here refers to a disease in which serum calcium levels are abnormally high, and it often co-occurs with the following diseases. For example, malignant tumor, vitamin D toxicity, sarcoidosis, hyperparathyroidism, etc. This substance is a known substance and was published by IKEKAWA, N.; Biochem.
Biophys.Res.Commun. 62 , 485, 1975. The substance may be 24R-(OH)-D ₃ , 24S-(OH)-D ₃ or a mixture thereof, but especially 24R-
(OH) _-D3 is preferred. The antihypercalcemic agent of the present invention can be used in various formulations as shown below, containing the above-mentioned substances as active ingredients. The antihypercalcemic agents of the present invention may be administered orally, parenterally or rectally, with oral administration being preferred. Preparations containing this substance as an active ingredient are used in dosage forms such as tablets, powders, granules, suppositories, capsules, alcoholic solutions, oily solutions, and aqueous suspensions. Further, as the oily solvent, intermediate fatty acid triglyceride ester, corn oil, cottonseed oil, peanut oil, fish liver oil, oily ester, etc. are used. Also preferred are cacao oil and glycerin. Other ingredients such as lactose, starch, talc, magnesium stearate, sorbic acid, salts of sorbic acid, sugar or alcohol derivatives thereof, physiological saline, surfactants, antioxidants, etc. may be used in combination with this substance. The substance may be present in a unit dosage form of 2 x 10 ^-6 to 2
×10 ⁻¹ % by weight, preferably 2×10 ⁻⁵ to 1×10 ⁻¹
% by weight. In addition, this substance is suitable for adults (average body weight)
1×10 ^-2 to 1× per day for 50 to 60 kg)
10 ⁴ μg, preferably 5×10 ⁻¹ to 3×10 ³ μg is administered. 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 triglyceride ester of intermediate fatty acids to a concentration of 0.1% and administered orally (po). The dose is 10mg/Kg. Observed for 1 week after administration
All 10 mice survived, and there was no difference in any way from the control group to which only triglyceride ester of intermediate fatty acid containing 0.1% ethanol was administered. Therefore, the LD ₅₀ value for oral administration of this substance is 10
It can be said to be extremely safe as it is more than mg/Kg. EXAMPLES The effects of the present invention will be specifically explained below with reference to Examples. Example 1 Vitamin _D3 dissolved in corn oil was orally administered to 16 groups of 5 male Wistar rats weighing 150±40 g for 4 consecutive days per 100 g body weight. Hypercalcemic rats were fasted overnight. The average serum calcium concentration of each rat before administration was 13.5±0.4 mg/dl. In the first group, 100μ of 24R-(OH) _-D3 dissolved in triglyceride ester of intermediate fatty acid was added.
g/Kg was administered orally, and the animals were sacrificed at 2, 5, 8, 12, and 24 hours later, and blood was collected from the abdominal descending vena cava.
OCPC (Orthocresol Phthalein Complexone)
Serum calcium levels were measured using the method. The second group was subcutaneously administered 4MRCH/Kg of porcine calcitonin dissolved in physiological saline containing 4% gelatin instead of this substance, treated in the same manner as the first group, and the serum calcium level was measured. The third group was used as a control, and only the triglyceride ester of intermediate fatty acid was administered, treated in the same manner as the first group, and the serum calcium level was measured. The results are shown in Table 1.

[Table] This substance lowered serum calcium levels to normal values when administered orally. And normal values were maintained for a long time. On the other hand, with porcine calcitonin, almost no effect was observed after 2 hours. Moreover, almost no effect was observed when the same substance was administered orally. Example 2 Each group consisted of 5 male Wistar rats weighing 150±40 g, and the rats in 4 groups were fasted all day and night. 1st
Groups were orally administered 100 μg/Kg of 24R-(OH)-D ₃ dissolved in triglyceride ester of intermediate fatty acids. The animals were sacrificed 2 and 5 hours after administration, blood was collected from the abdominal descending vena cava, and serum calcium levels were measured using the OCPC method. Similarly, a second group to which only triglyceride ester of intermediate fatty acid was administered was used as a control group, and serum calcium levels were measured in the same manner as above. The results are shown in Table 2. Administration of this substance did not change serum calcium levels.

[Table] Example 3 One group consisted of five male Wistar rats weighing 150±40 g, and the fourth group received parathyroid hormone (PTH) (Sigma) dissolved in physiological saline containing 4% gelatin.
Bovine parathyroid TCA powder134USPU/
mg) was subcutaneously administered at a dose of 150 USPU/0.4 ml per 100 g of body weight to create hypercalcemic rats. One hour after PTH administration, 50 μg/Kg of 24R-(OH)-D ₃ dissolved in ethanol was intraperitoneally administered to the first group. After 2 and 5 hours, the animals were sacrificed and blood was collected from the abdominal descending vena cava.
Serum calcium levels were measured using the OCPC method. As a second group (control group), only ethanol was administered intraperitoneally, and serum calcium levels were measured in the same manner as above.

[Table] Example 4 This example shows that this substance has the effect of preventing the onset of hypercalcemia caused by the administration of 1α-hydroxycholecalciferol (abbreviated as 1α-(OH)-D ₃ ). There were 6 groups of male Wistar rats weighing 130±30 g with 5 rats per group. After an overnight fast, the first group was given 10μg/1α-(OH) _-D3 dissolved in ethanol.
Kg was administered intraperitoneally. The second group received 10 μg/Kg of 1α-(OH)-D ₃ dissolved in ethanol intraperitoneally, and at the same time 100 μg/Kg of 24R-(OH)-D ₃ dissolved in triglyceride ester of intermediate fatty acids.
Administered orally. A group to which only intermediate fatty acid triglyceride ester was orally administered was defined as a control group. 10 hours after administration for each group and
After 18 hours, the animals were sacrificed, blood was collected from the abdominal descending vena cava, and the serum calcium level was measured according to the procedure described in Example 1. The results are shown in Table 4. As shown in Table 4, while 1α-(OH)-D ₃ increased serum calcium levels, when used in combination with 24R-(OH) ₂ -D ₃ , the increase in serum calcium levels was prevented. .

[Table] Example 5 Ascitic fluid from Walker carcino sarcoma 256 was injected 2x into the right upper thigh of 90-140gr Sprague-Dawley rats.
Inject 0.5 ml of the solution adjusted to 10 ⁶ tumor cells/μl, and add 1 ml of 24R-(OH)-D ₃ (3 μg/600 μl) dissolved in intermediate fatty acid triglyceride ester.
Orally administered at 20 μg/Kg per day for one week,
After sacrifice, blood was collected from the abdominal descending vena cava, serum calcium levels were measured according to a conventional method, and compared with a control group in which only triglyceride ester of intermediate fatty acids was administered. The results are shown in Table 5.

[Table] Example 6 1 kg of triglyceride ester of intermediate fatty acid
Dissolve _5mg of 24R-(OH)-D in 1 capsule.
The following shell components were heated and dissolved to contain 0.5 μg of 24R-(OH)-D ₃ and soft capsules were prepared using a soft capsule making machine in a conventional manner. Shell formulation example Gelatin 10 parts by weight Glycerin 4 Sorbic acid 0.1 Water 15 In the same way, capsules each containing 1 μg, 2 μg or 5 μg were prepared. next,
The capsules prepared as described above were orally administered to a 38-year-old female patient with hyperparathyroidism and hypercalcemia. 24R−
(OH)-D ₃ was administered continuously for 4 days at a dose of 10 μg/day. Next, blood was collected and the serum calcium level was measured using the same procedure as described in Example 1, which was 10.9 mg/dl.
I got a value of The serum calcium level before administration was 13.7 mg/dl. Example 7 _24R- (OH)-
_D3 was orally administered for 3 consecutive days. 24R−(OH)−D ₃
The dose was 10 μg/day. take blood,
The serum calcium level was measured using the same procedure as described in Example 1, and a value of 10.6 mg/dl was obtained. The serum calcium level before administration was 13.1 mg/dl. Example 8 A 35-year-old female patient diagnosed with sarcoidosis and associated with hypercalcemia received 24R-(OH)-D ₃ per day similar to that obtained in Example 6.
10 μg was orally administered for 4 consecutive days. Blood was collected on the 6th day, and the serum calcium level was determined using the method described in Example 1, and was found to be 10.6 mg/dl. The serum calcium level before administration was 13.3 mg/dl.

Claims (1)

[Claims] 1. An antihypercalcemic agent containing 24-hydroxycholecalciferol as an active ingredient. 2 24-hydroxycholecalciferol is 24R
- The antihypercalcemic agent according to claim 1, which is hydroxycholecalciferol.