JPH06239754A - Medicine for articular rheumatism - Google Patents

Abstract

PURPOSE:To obtain a medicine for articular rheumatism, having an excellent anti-articular rheumatism effect, low in toxicity and capable of effectively exhibiting its effect by a small dose. CONSTITUTION:This medicine for articular rheumatism is prepared by reacting gold or a gold compound with a nucleic acid (DNA and/or RNA) derived from a biological material and using the resultant gold-nucleic acid complex. A DNA or an RNA and gold or a gold compound are added to a prescribed buffer solution in a weight ratio of (DNA:gold) of (1:10) to (100:1) and allowed to react under stirring at 100 to 300 deg.C in an autoclave with a stirrer. After completion of the reaction, the resultant gold-nucleic acid complex is separated and purified according to the liquid chromatography method, the dialysis method, etc.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for rheumatoid arthritis, and more particularly to a therapeutic agent for rheumatoid arthritis containing a gold nucleic acid complex.

[0002]

2. Description of the Related Art In recent years, the number of patients with rheumatoid arthritis has been increasing, but the cause of its onset has not yet been clarified. Therefore, there is no causal therapy, and it is one of the intractable diseases. Currently, as therapeutic agents, anti-inflammatory agents, immunomodulators, immunosuppressants, etc. that act on the immune abnormalities that are the basis of inflammation are being used symptomatically.

Gold therapeutic agents such as gold colloidal solution, gold thiomaleate and gold thioglucose have been used as therapeutic agents that act on immune disorders, and they act on macrophages and exert their effects by changing the immune reaction. Are believed to be present. There are two types of gold preparations, parenteral and oral, which have good therapeutic effects, but they are slow-acting drugs that require 2-3 months to develop their effects, and dermatitis, rash, proteinuria, and gastrointestinal disorders. , Side effects such as stomatitis, leukopenia and pulmonary fibrosis. For this reason, it has been proposed to previously use gold-supported calcium apatite as a gold preparation with few side effects (JP-A-4-187638). Since calcium apatite is the main component of bone and is harmless to the human body, use of calcium-loaded apatite as an anti-rheumatic agent suppresses the occurrence of side effects,
Although effective, the oral absorption of calcium apatite is poor, and it may be necessary to take a large amount of oral doses to exert its effect.

[0004]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a rheumatoid arthritis agent which has an excellent anti-rheumatoid arthritis effect, is less toxic, and effectively acts even when administered in a small amount.

[0005]

[Means and Actions for Solving the Problems] As a result of various studies for solving the above problems, it was found that the problems can be solved by using a complex consisting of a nucleic acid and gold as an anti-rheumatoid arthritis drug. The nucleic acid used in the present invention is D
It contains NA and / or RNA and is extracted and collected by a conventional method from biological materials such as animals, plants, bacteria, and molds. DNA and RNA of animal origin include, for example, pigs, horses, cows, sheep, chickens,
For fish, for plant origin, for example, wood, vegetables, algae, and for bacterial origin, for example, lactic acid bacteria, Bacillus subtilis, Escherichia coli, brewer's yeast, rice koji mold, etc. it can.

That is, after suspending the biological material in Tris-HCl buffer, adding lysozyme and stirring for several tens of minutes,
Add additional protease and hold at 37 ° C for at least 1 hour. Add an equal amount of phenol, mix, centrifuge and collect the aqueous phase. Sodium acetate is added to the collected aqueous phase, and then double volume of cold ethanol is added to keep the temperature at -20 ° C for one day. The precipitate is collected by centrifugation and dried to obtain a mixture of DNA and RNA. It is possible to use this mixture as it is, but further to use DNase or RNa.
It is also possible to purify DNA or RNA for use by treating with se.

The DNA or RNA thus extracted is used to react with gold or a gold compound. D in a given buffer
NA or RNA and gold or a gold compound are put and reacted by applying temperature and pressure. The type of buffer used includes sodium phosphate, potassium phosphate, sodium acetate, sodium citrate, ammonium acetate, Tris-hydrochloric acid, Tris-phosphoric acid, etc., but other buffers can also be used. In addition to buffer solutions, hydrochloric acid, phosphoric acid, nitric acid, sodium hydroxide, calcium hydroxide, ammonia water, or salts such as sodium chloride, calcium chloride, ammonium chloride, urea can be added to these buffer solutions for reaction. Is. The concentration of the buffer solution may be between 1 mM and 10M. The buffer pH can range from 1 to 14. The kind and molecular weight of the nucleic acid that binds to gold can be arbitrarily adjusted by changing the concentration of buffer solution, pH, and added salts during synthesis.

The ratio of DNA and gold to the reaction solution may be between 0.001% and 50% by weight. D
The weight ratio of NA to gold may be between 1:10 and 100: 1. As the gold used, gold such as metallic gold or chloroauric acid can be used. Reaction temperature is 10
Perform at any temperature between 0 ° C. and 300 ° C. and synthesize using a pressure vessel with stirrer. For this reason the pressure is the water vapor pressure at the reaction temperature. The reaction time is such that the reaction is carried out for several hours or for several days immediately after reaching a predetermined temperature while stirring. After completion of the reaction, liquid chromatography or
The reaction product is separated and purified by dialysis or the like.

The nucleic acid gold complex according to the present invention is stable,
Conventional methods can be used for formulation. For example, in the case of an oral preparation, excipients commonly used in pharmaceuticals such as lactose, sucrose, starch, dextrin, crystalline cellulose,
Tablets, granules, capsules, pills or suspensions mixed with calcium phosphate, calcium carbonate, talc, magnesium carbonate, magnesium stearate, gelatin, etc.
As a preparation such as an emulsion, macrogol, glycerogelatin, cocoa butter and the like can be used as a suppository, or can be used as an injection by suspending or mixing with a solution such as distilled water or salt or potassium chloride. Hereinafter, the present invention will be described specifically with reference to Examples.However, since the gold nucleic acid compound was synthesized by the above method in all of the following Examples, the reaction conditions used in each Example are briefly shown in the table. It was

[0010]

[Example 1]

[Table 1]

[0011]

Example 2

[Table 2]

[0012]

Example 3

[Table 3]

[0013]

Example 4

[Table 4]

[0014]

Example 5

[Table 5]

[0015]

Example 6

[Table 6]

[0016]

Example 7

[Table 7]

[0017]

Example 8

[Table 8]

[0018]

[Example 9]

[Table 9]

[0019]

Example 10

[Table 10]

[0020]

Example 11 The antirheumatic effect of the gold compound synthesized as described above is shown. SD male rats having a body weight of 200 to 220 g were preliminarily bred for 1 week and then used in the experiment.
Under ether anesthesia, 0.3 mg / 0.05 ml of mycobacterium butyrican suspended in liquid paraffin was intradermally injected into the footpad of the right hind leg to prepare an adjuvant arthritis rat. Example 1 for 8 animals per group starting from 11 days, once a day for 30 days
0.04 mg / kg / day of the gold compound prepared in
The anti-rheumatic effect was judged by observing the condition of the joint. Similar tests were conducted for other compounds. For comparison, a group not administered with anti-rheumatic drug was used. After the final administration, the animals were dissected and observed for gastrointestinal disorders. Evaluation method 0: No change 1: Redness around the joint 2: Redness and swelling around the joint 3: Swelling and deformation of the joint The four limbs of the rat were observed, and each limb was graded from 0 to 3 according to the above-mentioned evaluation method. expressed. The maximum of one is 12 points. The obtained results are shown in Table 11.

[0021]

[Table 11]

As shown in the above results, a remarkably high anti-rheumatoid arthritis effect was recognized. As a result of dissection after the final administration and observation of gastrointestinal disorders, lesions such as erosion and ulcer on the mucous membrane of the stomach and intestines were not observed.

[0023]

INDUSTRIAL APPLICABILITY The gold-supported nucleic acid complex according to the present invention is considered to be useful as a therapeutic agent for rheumatoid arthritis, showing a remarkable anti-rheumatoid arthritis effect even when administered in an extremely small amount and no side effects.

Claims (1)

[Claims] 1. A rheumatoid arthritis agent comprising a complex consisting of a nucleic acid and gold.