JPH0223532B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to a prophylactic and therapeutic agent for collagen disease or rheumatoid arthritis. [Prior Art and Problems] Collagen disease and rheumatoid arthritis are already well known, but to date, no effective preventive or therapeutic agent has been provided. [Means for solving the problem] Through years of research, the present inventors have found that protease (proteolytic enzyme) and polysaccharide or mucopolysaccharide degrading enzyme are effective in preventing and treating chronic nephritis and chronic hepatitis. We found that this was the case and filed a patent application (Japanese Patent Application No. 125002/1982: Japanese Patent Application No. 26822/1983). This was the result of further research. Among proteases, Streptomyces
Neutral protease produced by the genus Serratia, Serratiopeptidase produced by the genus Serratia,
We discovered that thermolysin produced by the genus Bacillus is effective in preventing and treating collagen diseases such as progressive nasal gangrene and rheumatoid arthritis. I confirmed that there was no. Neutral protease produced by the genus Streptomyces, which is one of the active ingredients of the present invention, is obtained from a culture solution of Streptomyces griseus.
Ion exchange resins such as carboxymethyl (CM) cellulose and diethylaminoethyl (DEAE) cellulose, as well as carbobenzyloxyglycylleucine (Cbz-Gly-Leu), leucineamide (Leu-NH ₂ ), phenylalanine (Phe), etc. It is purified by chromatography using an affinity carrier (such as Sepharose) having as a ligand, and has the following chemical properties.

【table】

〔Example〕

Next, an example will be described. (Example 1) After uniformly mixing 2 g of serratiopeptidase derived from Serratia ST.E-15, 190 g of lactose, and 70 g of potato starch, a 3% hydroxypropyl cellulose aqueous solution is poured and kneaded. The mixture is pulverized, and the granules are mixed with 0.3% magnesium stearate and compressed into tablets. (Example 2) After mixing 2 g of neutral protease derived from Streptomyces griseus and 40 g of lactose, adding 10 g of hydroxypropyl methylcellulose and forming into granules,
Evenly coat with cellulose acetate phthalate to form enteric-coated granules. (Example 3) The enteric-coated neutral protease granules obtained in Example 2 are filled into capsules to prepare capsules. (Example 4) 6157 mg of Witepsol H-15 (manufactured by Dynamite Nobel Co., Ltd.; higher saturated fatty acid triglyceride) and 325 mg of polyoxyethylene lauryl ether (manufactured by Nikko Chemicals Co., Ltd.; BL-25) are added and melted by heating. To this, 500 mg of neutral protease powder produced by Streptomyces griseus is added, dispersed, and injected into a 130 mg suppository mold, and after solidification, separated from the mold. [Clinical case] Progressive nasal gangrene is the name of a necrotic granulation disease of the nasal cavity with a poor prognosis, but with advances in treatment methods in recent years, it is no longer necessarily hopeless, but the mortality rate is lower than that of cancer, sarcoma, and leukemia. It is an extremely malignant disease. (Clinical case 1) Female, 20 years old, office worker Diagnosis: Progressive nasal gangrene Chief complaints: Epistaxis, purulent rhinorrhea, fever. History of current illness: Approximately 6 months ago, a blood clot was observed in the nasal septum, and when the clot was removed, a perforation was found in the nasal septum. Gradually, the formation of eschar became obvious and the perforation gradually enlarged, resulting in a saddle nose after 2 months. Present symptoms: Skin pain, purpura, and ulcer formation were observed in both upper arms, both elbow joints, and both knee joints. The face was slightly edematous, the nose was slightly saddle-shaped, the nasal cavity had atrophy of the inferior turbinate mucosa on both sides, and the middle turbinate mucosa on both sides was covered with purulent bloody eschar, and a small amount of bleeding was observed when the eschar was removed. Treatment: Chief complaints such as fever, purulent odor, rhinorrhea, and epistaxis, medical history, sinus and lung radiographs, blood sediment value, r-
The above disease was diagnosed based on elevated globulin levels and intranasal findings, and steroid hormones (2 mg/day of Rinderone were administered for 2 months). Although the intranasal findings remained stable and were at the same level as at the time of admission, steroid side effects were observed. A full moon-like face, abdominal distension, skin striae, and band-like eruption in the crypt area were also observed, and renal function was slightly impaired.We diagnosed that the prognosis was the worst and would die if the situation continued, and steroids were discontinued, but the disease returned. The fever and purulent odor worsened, and rhinorrhea and occasional nosebleeds were observed. At this point, the patient took 6 neutral protease tablets a day and observed the progress. One month later, the fever and purulent odor had slightly improved. improved, 2
After a month, rhinorrhea decreased, nosebleeds disappeared, and after 4 months,
Although the condition was almost completely cured, he continued to take oral medication for two months. (Clinical case 2) Male, 51 years old Diagnosis: Progressive nasal gangrene Medical history: 4 years ago, the patient visited an internist complaining of high fever, but the fever did not subside, so he visited an outpatient clinic. Current symptoms: Foul-smelling rhinitis and swelling at the root of the nose were observed, and the nasal septum was covered with a small perforation and a blood clot. X
The patient was diagnosed with the above disease through radiographic examination and biopsy examination. In addition to general blood tests, tests for protein fraction, cholesterol, and immunoglobulin levels revealed that the patient was severely impaired. This patient temporarily recovered after receiving adrenocortical hormone therapy at another large hospital, but the disease relapsed and this time the same hormone therapy had little effect, so the dose was extremely large, and plasma The protein fraction was also severely impaired and the prognosis was diagnosed as poor. Treatment with neutral protease: Six tablets of neutral protease were administered per day, two tablets after each meal, and the progress was observed. One month later, the protein fraction, immunoglobulin, red blood cell count, and white blood cell count gradually improved. Two months later, the general condition gradually recovered, rhinorrhea decreased, and nosebleeds disappeared. Five months later, the patient was almost healed. , took oral medication for six months. (Clinical cases 3 and 4) Systemic lupus erythematosus 2 cases Female, 45 years old Female, 26 years old Six months after administering 6 neutral protease tablets a day,
The symptoms improved and there was no recurrence. (Clinical Cases 5 and 6) Scleroderma 2 cases Female, 56 years old, male, 48 years old The patient was treated with 6 neutral protease tablets a day and recovered in 6 months. The present inventor previously used Empinase P in a case of collagen cachexia, and said that the administration of 6 tablets per day had a positive effect on the substances secreted from the lesion. We developed a successful treatment for the following diseases: (1) progressive nasal gangrene, (2) systemic lupus erythematosus, and (3) scleroderma by administering neutral protease. (Clinical Case 7) Female, 64 years old Diagnosis: Chronic rheumatoid arthritis, osteoarthritis Chief complaint: Right knee joint pain. History of current illness: Diagnosed with rheumatoid arthritis about 20 years ago and has been receiving treatment. Six years ago, the patient developed intermittent lameness of the cauda equina due to degenerative spinal stenosis. One year ago, I started experiencing walking pain on the inside of my right knee joint. Then, the patient's right knee joint gradually began to swell, making it difficult to carry out activities of daily living. Current symptoms: The patient had an obese physique, swelling of the right knee joint, and tenderness in the joint space on the medial side of the right knee joint. Unable to move or reach maximum extension of right knee, extension: -15 degrees, flexion: 135 degrees
It was hot. X-ray findings: There was bone spur formation on the inside of the right knee joint.
The joint space was narrow. Treatment with neutral protease: Six neutral protease tablets were taken per day, two tablets after each meal. Two months later, the right joint pain disappeared, joint swelling and osteophyte formation were alleviated, and the joint space expanded slightly. [Animal test example] The anti-inflammatory effect of enzymes was investigated. Average weight 170g
Pleurisy was induced by administering 0.1 ml/rat of 2% carrageenan into the pleural cavity of the rat, and 5 hours later, the amount of exudate in the pleural cavity, total protein content, and total cell number, which are indicators of pleurisy, were measured. The enzymes were administered intravenously at their respective doses 2 minutes before the administration of carrageenan.
The results are shown in Table 1. Enzyme administration improved pleuritis in a dose-dependent manner. Also, 0.3
A stronger improving effect was observed with mg/Kg of the enzyme than with 1 mg/Kg of pronase, confirming that this enzyme has a stronger anti-inflammatory effect than pronase.

[Test Example 1-Oral administration]

General symptoms: In the 4000mg/Kg and 2000mg/Kg groups,
5 to 15 minutes after administration, some animals showed suppression of locomotor activity and crepitus breathing sounds, watery stools were also observed 1 hour after administration, and 3 to 5 hours after administration, all animals of both sexes in the 4000 mg/Kg group. All males and 3 females in the 2000 mg/Kg group, the next day.
One female in the 2000 mg/Kg group died. In the surviving cases, no noteworthy symptoms were observed on the day after administration. In the 1000 mg/Kg group, suppression of locomotor activity and watery stools were observed from 1 hour after administration, and one female died after 5 hours, but in the surviving cases, symptoms were alleviated after 3 hours. It disappeared the next day. Based on the above mortality situation, the LD50 value is 1000.
It was estimated to be between ~2000mg/Kg. Body weight: No effect on body weight was observed. Pathological examination: In the case of death, bleeding in the kidney medulla, the entire gastrointestinal tract, and the lungs was observed macroscopically, and histological findings confirmed the macroscopic findings, as well as necrosis and sloughing of the intestinal mucosal epithelium and 4000 mg/Kg group. Hemorrhage in the cerebral choroid plexus was observed in the male. No abnormal findings were observed in the surviving cases. [Test Example 2 - Intravenous administration] General symptoms: In the 20 mg/Kg group, suppression of spontaneous movement, prone position, slow to difficult breathing, lacrimation, salivation, and swelling of the auricles and quadrats occurred within 5 to 15 minutes after administration. Symptoms such as cyanosis and tail congestion were observed, and all cases died. Ten
In the mg/Kg group, symptoms similar to those in the 20 mg/Kg group gradually developed, and all cases died 15 minutes to 5 hours after administration. In the 5 mg/Kg group, only bleeding in the tail was observed 5 to 15 minutes after administration. Body weight: No effect on body weight was observed. Pathological examination: In the case of death, hemorrhage was observed macroscopically in the kidney medulla, the entire gastrointestinal tract, and the lungs, and histological findings confirmed the macroscopic findings, as well as necrosis and sloughing of the intestinal mucosal epithelium and cerebral choroid plexus. Bleeding was observed. No abnormal findings were observed in the surviving cases. (Appendix 2,
3. As shown in the attached figure).

【table】

【table】

〔Effect of the invention〕

The prophylactic and therapeutic agent for collagen disease or rheumatoid arthritis of the present invention contains only a neutral protease selected from neutral protease produced by Streptomyces, Serratiopeptidase produced by Serratia, and thermolysin produced by Bacillus, or The neutral protease and other proteases migrate to the injured area in extremely large amounts, and disappearance of edema, infiltration of round cells, neovascularization, and fibrinolysis are observed in the tissue of the injured area, leading to collagen disease. It also has the effect of improving the chronic inflammation symptoms of chronic arthritis.

Claims (1)

[Claims] 1. A prophylactic and therapeutic agent for collagen disease or rheumatoid arthritis, which contains as an active ingredient a neutral protease selected from neutral protease produced by Streptomyces, Serratiopeptidase produced by Serratia, and thermolysin produced by Bacillus.