JPS6221329B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an antihypercalcemic agent containing 24,25-dihydroxycholecalciferol. Calcitonin has conventionally been used as a therapeutic agent for hypercalcemia. However, calcitonin, which is a peptide hormone, has a short lifespan in the body, and there are problems with the sustainability of its effects. Furthermore, since calcitonin cannot be administered orally, it must be administered by intramuscular injection or the like, which has the disadvantage of causing pain to the patient. Furthermore, it has been pointed out that administering calcitonin of non-human origin is associated with risks such as decreased efficacy due to antibody production and anaphylactic shock. In view of the above-mentioned facts, there has been a long-awaited development of an antihypercalcemic agent that has excellent long-lasting pharmacological effects and can be administered orally. As a result of intensive research into a substance that is endogenous to healthy humans and has been proven to be safe, the present inventors found that 24,25-dihydroxycholecalciferol (hereinafter referred to as this substance or 24,25-dihydroxycholecalciferol)
The present invention was achieved based on the knowledge that (OH) ₂ -D ₃ ) is effective in terms of long-lasting pharmacological effects and oral administration. All of these substances are known substances and have the following structures, for example, Pharmacia, 10 : 319-322, 1974.
has been disclosed. The effect of this substance on serum calcium is described in the following literature: (a) JMCanterburg: J.Clin.Invest.; 78 , 1375
~1383, 1978. When 24·25−(OH) ₂ −D ₃ was continuously injected into the thyroid artery of a normal dog over a long period of time (20 minutes),
It has been reported that arterial administration of 24·25-(OH) ₂ -D ₃ causes a temporary decrease in normal serum calcium levels, which quickly return to normal values. Incidentally, continuous administration into the thyroid artery is a special administration method, unlike the usual oral, subcutaneous, intraperitoneal, intravenous, etc. administration. (b) JMCanterburg; Clin.Res.; 26 , N1;
PA31, 1978. When 24·25-(OH) ₂ -D ₃ was administered to dogs with hyperepithelial function via the thyroid artery route, it was reported that the calcium level did not change. This report is written by the same person as the author of document (a) above. (c) AW Norman; Brit.Med.J.; 280 (No.
6212), 449-450, 1980. When 250 μg of 24·25-(OH) ₂ -D ₃ was orally administered to normal subjects, it was reported that the serum calcium level did not change significantly. (d) Others, JAKanis, Brit.Med.J. (1978)
1 1382-1386; H. Rasmussen, J. Clin.
Endocrinol. Metab. 46 284-294 (1978); J.
Szymendera, Brit.Med.J. (Nov.28). 1465~
1466 (1978) and JAKanis.V-D Basic
Research and its Clinical Application
(1979) 119-122. However, in all of them, 24-25-
There are no reports of cases in which administration of (OH) ₂ -D ₃ significantly changes serum calcium levels. As mentioned above, there are no reports of administration of 24·25-(OH) ₂ -D ₃ to animals with hypercalcemia or the same condition. The present inventors have discovered that 24·25-(OH) ₂ -D ₃ restores abnormally elevated serum calcium levels to normal levels, and maintains its effect for a long period of time. Therefore, this substance can be said to be extremely useful as an antihypercalcemic agent. Hypercalcemia here refers to a disease in which serum calcium levels are abnormally high, and is often associated with the following diseases. For example, malignant tumors, vitamin D
Poisoning, sarcoidosis, hyperparathyroidism, etc. This substance is 24R・25−(OH) ₂ −D ₃ , 24S・25−
(OH) ₂ -D ₃ or a mixture thereof may be used, but 24R.25-(OH) ₂ -D ₃ is particularly 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. As the oily solvent, triglyceride esters of intermediate fatty acids, corn oil, cottonseed oil, peanut oil, fish liver oil, oily esters, 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 its derivative alcohol, physiological saline, surfactants, antioxidants, etc. may be used in combination with this substance. This substance may contain 0.000002 to 0.4 in unit dosage form.
% by weight, preferably 0.00002 to 0.1% by weight. In addition, this substance is 0.01 μg per day for adults.
Administer 10,000 μg, preferably 0.5-300 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 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. In addition, 24R and 25 used in the examples
Structure confirmation of the optical isomer at position 24 of −(OH) ₂ −D ₃ is
Tetrahedron Letters No.26, pp2203-2206,
This was done with reference to 1975. 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 at 300r/0.5ml per 100g 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, 24R・25−(OH) ₂ −D ₃ dissolved in triglyceride ester of intermediate fatty acid was added.
100 μg/Kg was administered orally, and the animals were sacrificed after 2, 5, 8, and 12 hours, 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 4 MRCU/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 triglyceride ester of intermediate fatty acid was administered, and the group was 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
100μg/2R4・25-(OH) ₂ - _D3 dissolved in triglyceride ester of intermediate fatty acid was added to the group.
Kg was administered orally. 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 powder 134 USPU／
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·25-(OH) ₂ -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 administration of 1α-hydroxycholecalciferol (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 received 100 μg of 24R·25-(OH) ₂ -D ₃ dissolved in triglyceride ester of intermediate fatty acids.
g/Kg was administered orally. In addition, only triglyceride ester of intermediate fatty acid was orally administered to the control group. Animals in each group were sacrificed 10 and 18 hours after administration, blood was collected from the abdominal descending vena cava, and serum calcium levels were measured according to the procedure described in Example 1. The results are shown in Table 4. As seen in Table 4, 1α-(OH) _-D3 increases serum calcium levels, while 24R・25-
When (OH) ₂ -D ₃ was used in combination, 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 24R・25-(OH) ₂ -D ₃ (3 μg/600 μ) dissolved in intermediate fatty acid triglyceride ester adjusted to 10 ⁶ tumor cells/ml.
was orally administered at 20 μg/Kg per day for one week continuously, blood was collected from the abdominal descending vena cava after sacrifice, and serum calcium levels were measured according to the usual method. compared.

[Table] Example 6 1 kg of triglyceride ester of intermediate fatty acid
Dissolve 5 mg of 24R・25−(OH) ₂ −D ₃ and heat and dissolve the skin components below so that each capsule contains 0.5 μg of 24R・25−(OH) ₂ −D ₃ to produce soft capsules. Soft capsules were prepared using a conventional method using a machine. Example of shell formulation 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 40-year-old male patient with hyperparathyroidism and hypercalcemia. 24R・25
-(OH) ₂ -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.
I got the value dl. The serum calcium level before administration was 13.8 mg/dl. Example 7 The results obtained in Example 6 were obtained in a 58-year-old male patient who underwent hemodialysis due to renal failure and developed hypercalcemia after taking active vitamin D ₃ orally at 0.5 μg to 1.0 μg/day for 4 months. Capsules similar to those were orally administered for 3 consecutive days. The dosage of 24R・25−(OH) ₂ −D ₃ is 10μ
g/day. Blood was collected and the serum calcium level was measured using the same procedure as described in Example 1.
A value of mg/dl was obtained. The serum calcium level before administration was 12.8 mg/dl. Example 8 A 32-year-old female patient diagnosed with sarcoidosis and accompanied by hypercalcemia received 5μ of 24R·25-(OH) ₂ -D ₃ similar to that obtained in Example 6.
Two capsules containing g were orally administered per day for 5 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 10.8 mg/dl.
The serum calcium level before administration was 14.0 mg/dl.

Claims (1)

[Claims] 1. An antihypercalcemic agent containing 24,25-dihydroxycholecalciferol as an active ingredient. 2. The antihypercalcemic agent according to claim 1, wherein the 24,25-dihydroxycholecalciferol is 24R,25-dihydroxycholecalciferol.