JPS58109421A - Agent for preventing immunodeficiency - Google Patents

Abstract

PURPOSE:To prepare the titled agent containing 24,25-dihydroxycholecalciferol as an active component, and effective to the immunodeficiency of various renal diseases such as chronic renal insufficiency, uremia, nephrotic syndrome, etc. CONSTITUTION:A composition containing 24,25-dihydroxycholecalciferol[abbreviated as 24,25-(OH)2D3]as a main component is used as an agent for preventing the immunodeficiency of renal diseases. The above 24,25-(OH)2D3 includes 24R,25-(OH)2D3. 24S,25-(OH)2D3, and their mixture, however, 24R,25- (OH)2D3 is especially preferable. In the immunoinsufficiency in renal diseases, the cellular immunoinsufficiency is more remarkable than humoral insufficiency. The influence of the compound to the thymus gland which mainly manages the cellular immune function was studied, and it has been found that the lowering of the weight ratio of thymus/body, the number of nucleus thymic cells, the size pattern of the thymic cells, and the lymphocyte transformation can be prevented by the administration of said compound.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an agent for preventing decline in immune function. More details L< is 2
4,25-dihydroxycholecalciferol (hereinafter 2
4.25-(OH)tDs), the present invention relates to an agent for preventing immune function decline in renal diseases.

Renal failure and uremia are representative diseases that cause progressive and widespread immunodeficiency. In a recent national survey,
38.3% during acute renal failure and 19.3% during chronic renal failure.
2% infections were confirmed. Therefore, there are many deaths due to infectious diseases, and in long-term dialysis patient death statistics, infectious diseases account for 13.3%.
occupies . It is said that the immune dysfunction associated with renal failure is more markedly due to cell-mediated immune dysfunction than humoral dysfunction. (Clinical Immunology 12(8), 607-6
14 (1980)).

Therefore, by preventing or reversing the decline in immune function caused by kidney diseases, it becomes possible to prevent infections caused by various kidney diseases.

The present inventors conducted intensive studies on agents for preventing decline in immune function related to kidney diseases, and found that 24R125-(OH)!
The inventors discovered that Da is effective and arrived at the present invention.

24,25- (OH)2Da is USP371534
, H.F., DeLuca.

Biochemistry, 12(24), 485
1-55 ('73).

24.25-(OH)tDske24R of the present invention,
25-(OH)mDa.

It includes 24S, 25-(OH)*Da, or a mixture of both (hereinafter abbreviated as the present compound), with 24R,25-(OH\D being particularly preferred).

Cell-mediated immune dysfunction is more pronounced than humoral dysfunction in renal disease, so this compound's effects on the gut gland, which controls cell-mediated immune function, are mainly determined by Vc.
pl is solid. By administering the present compound, prevention of decreases in the thymus/body weight ratio, the number of nucleated thymus cells, the size pattern of thymocytes, and the juvenile response of lymphocytes was observed.

This compound is useful as an agent for preventing decline in immune function due to renal disease.

・For example, it is useful for immunodeficiency caused by various kidney diseases such as chronic renal failure, uremia, and nephrotic syndrome.

When the present compound is used as an agent for preventing decline in immune function, it is administered in a desired form of a millet composition according to a conventional method.

These may be administered orally, rectally or by other parenteral routes.

Preparations containing this compound as an active ingredient are tablets and powders.

Granules, suppositories, capsules, alcohol solutions.

It is used in dosage forms such as oily solutions and aqueous suspensions. Oil-based solvents include triglyceride esters of intermediate fatty acids, corn oil, and cottonseed oil.

Peanut oil, fish liver oil, aburo ester, etc. are used. Also preferred are cacao oil and glycerin.

Other ingredients include lactose, starch, and talc.

Stearin, magnesium acid, sorbic acid, salts of sorbic acid, sugars or derivatives thereof alcohols, seratin, physiological saline, surfactants, antioxidants, etc. may be used in combination with the present compound.1 The present compound may be used in unit dosage form. Inside 01100002~0
It may be contained in a quadruple column, preferably 0.00002 to 0.1% by weight. In addition, this compound is 0.0 per 71 days for adults.
1 pN~l 0711, preferably 0.1~300μI
Administer. Part 1. As a composition containing an amount of lever l
E:I oysters will be administered in 3 divided doses.

Next, we investigated the acute toxicity of this compound and the results are shown below.

! 81. Sexual toxicity: This compound was dissolved in ethanol using 10 ddN male medium mice (weight 20+3#), and the ethanol level was 01.
% in triglyceride ester of intermediate fatty acids and administered orally (p.

0) Administered. The dose was 1Orn9/kg. All 10 mice were observed for one week after the membrane formation, and there was no difference in any way from the control group to which only triglyceride ester of intermediate fatty acid containing 0.1 titanol was administered. Therefore, the LDIlo value of this compound for oral administration is 1011 anchors/kg or more, and it can be said to be extremely safe.

The present invention will be specifically described below by way of examples.

In addition, used in the examples [7ta24R,25(OH)gD
s, 24S, 25 (()t()gDs of 24
Tetrahedron is used to confirm the structure of optical isomers.
Letters, N026.

2203-2206 (/75).

Example 1 Thymus total cell number/Thymus weight ratio per body weight 10 week old male J
CL-Wistar rats were kept at a temperature of 21±lC1 and a humidity of 60
The animals were allowed free access to chow (CF2) and drinking water in a room containing ±5%.

The experiment was conducted in the following four groups, with eight animals in each group.

■Group physiological saline (subcutaneous administration) +MCT (oral administration)
Group II 7v+-ts++%y (subcutaneous administration) +MCT
(Oral administration) Group 1H AN-1 hzujun (subcutaneous administration) 124 25-(0H) t D, -100pgA (oral administration) Group IV AN-1 sil 屹~ (subcutaneous administration) +24a 2
5-(OH), DI-1o-Si (oral administration) A
N: )'++romyc in red nonucleo
side i Dissolved in saline.

tvFcT:C8-,. triglyceride ester of carboxylic acid.

24R, 25-(01()2 o3 knee 1.4cTr Nagisa pj Rei) Each group was administered continuously for 12 days, and the thymus was taken out from the abdomen and descending vena cava, and the attached IJ was taken out.
After removing the W jiff tissue, etc., it was immediately weighed on a microbalance. To determine the number of nucleated cells in the thymus, homogeneously loosen the thymus in phosphate buffered saline (1) with a bottle set,
After thorough pipetting and passing through a stainless steel mesh (#200), the tubes were stained with 'l''ur 1.c solution and counted under !# microscopy.

The results are shown in one table.

Table 1 Example 2 (Same experiment as Example 1) Measurement of cell size pattern using FAC8 Analysis of sebaceous gland cell size pattern using Becton-[)ic
Fluoresce-nca7 manufactured by lcinson
tivated Ce1lSorter (FACS
II) It was carried out using 'fr. Extracted breasts!
Loosen the il uniformly with tweezers in PBS, pipette thoroughly, and use a stainless mesh (4#)
200)? After passage, the cells were treated with a Tris-human xyamine methane buffered 0.83% ammonium chloride solution to hemolyze the mixed red blood cells, and further washed three times with PR8VC. The thymocytes thus obtained were incubated 5X in PBS.
FA(,
Cell size was analyzed by STI. The proportion of viable cells among the thymocytes thus obtained was confirmed to be 95% or more by staining with Trivan blue.

As a result of the measurement, the proportion of cells slightly larger than normal increased due to AN administration, 24.25-(OH')R1').

The treated group recovered to the same pattern as normal.

Example 3 Young generation reaction of peripheral blood lymphocytes The young generation reaction of peripheral blood lymphocytes was measured by the following method.

Hepa-11'1 venous blood was mixed with the same amount of RPMI-1640 medium, centrifuged in Ficoll-Paque solution for 20 minutes at 400XN overlay, and the mononuclear cell layer was fractionated and RPMI-1640 medium was mixed. 1640 medium, 3
RPMI with 10% calf serum added to cells washed twice
-1640 medium Lb IC1,5X 10' :l/
It was suspended in lnl Tonaruyo'). 100p of this suspension
1 Gold Falcon Micro Test ■Put into the recess of the plate, add 1 fine bubble mitogen (hereinafter referred to as mitogen), and add 101 calf serum to RPMI-164.
Add 0 medium to make the total volume 200 pl, and incubate in 37C air:carbon dioxide (95:5) water vapor saturated atmosphere.
7'r, 0 24 hours before the end of incubation sH
-Methylthymidine (specific activity 1g 5C1/mrn01)
0.5 μCi of was added.

After the incubation, one cell was taken out using a glass fiber, and the amount of isotope-J incorporated into the cell was measured using a liquid scintillation counter. In the above, ConA (concanavalin A)'& B-thin Jm mitogen as T cell mitogen: LP
S (lipopolysaccharide) was used and incubated for 72 hours.

The results are shown in Table 2.

(Group: Same as Example 1) Example 4 Normal human fresh peripheral blood (-valine m) WoFioll-
The lymphocyte layer was collected by centrifugation using Paque.

After washing the lymphocytes three times with PBS, RPMI-1640
The cells were adjusted to 6 x 10' cells/ynl using a medium.

The above lymphocyte solution is 170μ and the mitogen solution is 10μ!
After culturing the blood of the subject in a 20 μm, round-bottomed 96-well plate with a total volume of 200 pi for 65 hours,
1 μCi of “H-Tdr solution (IQ OpCi/ml
) was added, and after culturing for 10 hours, the cells were harvested and the uptake of lymphocytes (H-Tdr value f) was measured.

Patient's blood sputum is 24ft, 25-(on) for patients who are hospitalized or hospitalized due to chronic renal failure, urinary tract, etc. [).

A total of 0.5 to 10 p1 per day.Adults were divided into patients who received the drug for 1 to 6 months (n=8) and groups who did not (n=8), and blood samples were collected before and after dialysis. The results are shown in Table 3.

Table 3 Note: PHA = kidney bean lectin LPS = lipopolysaccharide cpm = count/= Example 5 Impure peroxide present was eliminated by irradiation with a high-pressure mercury lamp for 48 hours in an inert gas atmosphere.

Triglyceride ester of intermediate fatty acid l Ky to 24
1ζ, 25-(OH), D, 5~ is dissolved in 1 capsule, 24 R, 25-(OH)*Ds t O-5
μ? Soft capsules were prepared by heating and dissolving the following shell components so as to contain the following ingredients, and using a soft capsule making machine in a conventional manner.

Shell formulation example Gelatin 10 parts by weight Glycerin 41 Sorbic acid 0.1 parts by weight Water 15 N Similarly, 1μ in 1 capsule? , 2μ? or 58rr
Hemiya 1) Yutaka Hiro Continued from page 1 0 Inventor: Kato Kato 3-27K-40 Nakadai, Itabashi-ku, Tokyo ■Inventor: Yoshikumi Chikao 2-19-46 Kunitachi Shajo 2-19-46 Procedural amendment Commissioner of the Patent Office Shima 1) Haruki Tono1, Indication of the case: 1982 Patent Application No. 1980162, Name of the invention: Agent for preventing decline in immune function 4, Implantation: Yamada Building r O, 1-1-14 Shinjuku, Shinjuku-ku, Tokyo. 01μ2～10μm” should be changed to “001μm～
Correct it to 10m%J.

139-

Claims (1)

[Scope of Claims] (1) An agent for preventing decline in immune function containing 24,25-dihydroxycholecalciferol as a main component. <2) 24.25-Dihydroxycholecalciferol is 24R,25-dihydroxycholecalciferol, The agent for preventing decline in immune function according to claim (1). (3) The agent for preventing decline in immune function according to claim (1) or (2), which is used for renal diseases.