JP2017014204A - Improvement or treatment agent of chondromalacia patella - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a useful pharmaceutical agent for improving or treating chondromalacia patella.SOLUTION: The present invention relates to an improvement or treatment agent for chondromalacia patella comprising an extract of an inflammatory tissue inoculated with vaccinia virus as an active ingredient. A pharmaceutical agent containing the extract as an active ingredient is extremely useful because it has been found that the improvement or treatment effect of chondromalacia patella is high in humans and safety is also very high.SELECTED DRAWING: None

Description

  The present invention relates to a novel pharmaceutical use of an extract of inflamed tissue inoculated with vaccinia virus (hereinafter sometimes referred to as “the present extract”). More specifically, the present invention relates to an agent for improving or treating patella cartilage softening comprising the present extract as an active ingredient.

  Patellar chondrosis is a bone joint disease commonly seen in clinical practice, where the cartilage on the back of the patella softens and the pain of the patellofemoral joint is the main symptom. Many patients with patella chondrosis have a clear history of trauma or chronic microinjuries, and pathological features include softening, cracking, shedding, and degeneration of patella cartilage. The characteristic of arthralgia due to patella chondrosis is that it begins with dull pain and discomfort on the front side of the knee joint, and gradually develops into knee joint pain, especially when exercise such as climbing up and down stairs is severe. .

  Patients with moderate patella chondrosis are treated with oral nonsteroidal anti-inflammatory drugs and physical therapy. However, long-term physical therapies interfere with work and are expensive, so patient compliance tends to be low. In addition, since most patients refuse surgery, it is common practice to choose intra-articular injection therapy with hyaluronic acid preparations. Intra-articular injection therapy is widely used in clinical settings because it is effective and has few complications. Under such circumstances, a method for further improving the effect of intra-articular injection therapy is required in the clinical field.

  About the extract of inflammatory tissue inoculated with vaccinia virus, which is an active ingredient for improving or treating patella cartilage softening according to the present invention, analgesic action, sedative action, anti-stress action, anti-allergic action (see Patent Document 1), immune promotion Action, anticancer action, liver cirrhosis inhibitory action (see Patent Document 2), therapeutic effect on idiopathic thrombocytopenic purpura (see Patent Document 3), postherpetic neuralgia, cerebral edema, dementia, spinocerebellar degeneration etc. Therapeutic effect (see Patent Document 4), Raynaud's syndrome, diabetic neuropathy, therapeutic effect on SMON sequelae (see Patent Document 5), kallikrein production inhibitory action, peripheral circulation disorder improving action (see Patent Document 6), bone atrophy Improvement action (see Patent Document 7), Nitric oxide production inhibitory action (see Patent Document 8) effective for treatment of sepsis and endotoxin shock, therapeutic effect on osteoporosis Patent Document 9), AIDS treatment effect based on Nef action inhibitory action and chemokine production inhibitory action (see Patent Documents 10 and 11), therapeutic effect on ischemic diseases such as cerebral infarction (see Patent Document 12), fibromyalgia (See Patent Document 13), therapeutic effect on infectious diseases (see Patent Document 14), prevention or alleviation of peripheral neuropathy by anticancer agents (see Patent Document 15), chronic prostatitis, interstitial cystitis and / or Or the therapeutic effect of dysuria (see Patent Document 16), the production promoting action of neurotrophic factors such as BDNF (see Patent Document 17), and the collagen and proteoglycan synthesis promoting action in chondrocytes (see Patent Document 18). Wide range of actions and effects are known. However, it has not been known so far that this extract is effective in improving or treating patella cartilage softening.

JP-A-53-101515 JP 55-87724 A Japanese Patent Laid-Open No. 1-265028 JP-A-1-319422 JP-A-2-28119 JP-A-7-97336 JP-A-8-291077 JP-A-10-194978 Japanese Patent Laid-Open No. 11-80005 JP-A-11-139777 JP 2000-336034 A Japanese Unexamined Patent Publication No. 2000-16694 International Publication WO 2004/039343 JP 2004-300146 A International Publication WO2009 / 028605 International Publication WO2011 / 111770 International Publication WO2011 / 162317 International Publication WO2012 / 051173

  An object of the present invention is to provide a drug that is effective and effective in improving or treating patella cartilage softening.

  As a result of diligent research on pharmaceutical treatment of patella cartilage softening for which an effective treatment method is required, the present inventors have found that a preparation containing this extract as an active ingredient is excellent for patella cartilage softening The present invention was completed by finding that it shows an improvement or therapeutic effect.

  This extract has an excellent pharmacological action of improving or treating patella cartilage softening. Moreover, since the chemical | medical agent containing this extract as an active ingredient is a highly safe chemical | medical agent with few problems, such as a side effect, this invention is very useful.

  This extract is an extract containing a non-protein active substance extracted and separated from an inflamed tissue of an animal that has been inoculated with vaccinia virus. The extract is liquid in the extracted state, but can also be made solid by drying. A drug containing this extract as an active ingredient (hereinafter referred to as “the present preparation”) is very useful as a pharmaceutical product. In this case, since this extract is an active ingredient of this formulation, this extract is the drug substance of this formulation. A specific product manufactured and sold by the applicant in Japan as this preparation is “Vaccinia virus-inoculated rabbit inflammation skin extract-containing preparation” (trade name: Neurotropin / NEUROTROPIN (registered trademark)). This preparation includes injections and tablets, both of which are ethical drugs.

  The indications for injections of neurotropin are “back pain, neck-shoulder syndrome, symptomatic neuralgia, pruritus associated with skin diseases (eczema, dermatitis, urticaria), allergic rhinitis, cold feeling of SMON aftereffects. "Abnormal perception / pain". The indications for neurotropin tablets are “postherpetic neuralgia, low back pain, cervico-arm syndrome, peri-shoulderitis, osteoarthritis”. This formulation was created by the applicant and developed as a pharmaceutical product. Its superior efficacy and safety features have been evaluated, and it has been sold for many years, establishing a firm position in the Japanese pharmaceutical market. Is.

The inflamed tissue extract inoculated with vaccinia virus in the present invention crushes inflamed tissue that has been inoculated with vaccinia virus, and after removing tissue fragments by adding an extraction solvent, the protein is removed and adsorbed by the adsorbent And then eluting the active ingredient. That is, for example, the following steps.
(A) Collect skin tissues such as rabbits and mice that have been inoculated with vaccinia virus, crush the germed tissues, and add an extraction solvent such as water, phenol water, physiological saline, or phenol-glycerin water. After that, an extract (filtrate or supernatant) is obtained by filtration or centrifugation.
(B) The extract is adjusted to an acidic pH, heated and deproteinized. Next, the deproteinized solution is adjusted to be alkaline and heated, followed by filtration or centrifugation.
(C) The obtained filtrate or supernatant is acidified and adsorbed on an adsorbent such as activated carbon or kaolin.
(D) An extract from inflamed tissue inoculated with vaccinia virus can be obtained by adding an extraction solvent such as water to the adsorbent, adjusting the pH to alkaline, and eluting the adsorbed components. Thereafter, if necessary, the eluate can be made into a dried product by evaporating to dryness or freeze-drying under reduced pressure.

  As animals for inoculating vaccinia virus to obtain inflamed tissues, various animals infected with vaccinia virus such as rabbits, cows, horses, sheep, goats, monkeys, rats and mice can be used. Inflamed skin tissue is preferred. Any rabbit may be used as long as it belongs to the order of rabbit eyes. Examples include rabbits, rabbits (rabbits made from rabbits), rabbits (Japanese rabbits), rabbits, snow rabbits, and the like. Of these, chira rabbits are suitable for use. In Japan, there is a so-called rabbit that has been bred from the past and used widely as domestic animals or experimental animals. There are many varieties (breeds) of the rabbit, but varieties such as Japanese white varieties and New Zealand white varieties (New Zealand white) can be suitably used.

  The vaccinia virus may be of any strain. Examples include Lister strains, Daren strains, Ikeda strains, EM-63 strains, New York City Board of Health strains, and the like.

As a basic extraction process of the present extract, for example, the following processes are used.
Step (A) Inflammated skin tissue obtained by intradermal inoculation of vaccinia virus into the skin of rabbits is collected. The collected skin tissue is washed and disinfected with a phenol solution. This inflamed skin tissue is crushed and 1 to 5 times the amount of extraction solvent is added. Here, crushing means crushing finely into a mince using a mincing machine or the like. In addition, as extraction solvents, distilled water, physiological saline, weakly acidic to weakly basic buffer solutions, and the like can be used. Sterilizers and preservatives such as phenol, stabilizers such as glycerin, sodium chloride, potassium chloride Further, salts such as magnesium chloride may be appropriately added. At this time, extraction can be facilitated by disrupting the cell tissue by treatment with freeze-thaw, ultrasound, cell membrane lytic enzyme, surfactant or the like. The resulting suspension is left for 5 to 12 days. Meanwhile, the mixture may be heated to 30 to 45 ° C. with or without stirring as appropriate. The obtained liquid is subjected to solid-liquid separation (filtration, centrifugation, etc.) to remove the tissue piece to obtain a crude extract (filtrate or supernatant).

About a process (B) A protein removal process is performed about the crude extract obtained at the process (A). Deproteinization can be carried out by known methods commonly used, such as heat treatment, treatment with a protein denaturant (for example, an organic solvent such as acid, base, urea, guanidine, acetone, etc.), isoelectric precipitation, salting out. Etc. can be applied. Subsequently, the insoluble protein that has been precipitated by a usual method for removing insoluble matters, for example, filtration using filter paper (cellulose, nitrocellulose, etc.), glass filter, celite, zeit filtration plate, etc., ultrafiltration, centrifugation, etc. A removed filtrate or supernatant is obtained.

About Step (C) The filtrate or supernatant obtained in Step (B) is adjusted to acidic, preferably pH 3.5 to 5.5, and the adsorption operation to the adsorbent is performed. Examples of usable adsorbents include activated carbon, kaolin, and the like. Add the adsorbent to the extract and stir, or pass the extract through an adsorbent-filled column to add the active ingredient to the adsorbent. Can be adsorbed. When an adsorbent is added to the extract, the adsorbent can be obtained by adsorbing the active ingredient by removing the solution by filtration or centrifugation.

About step (D) In order to elute (desorb) the active ingredient from the adsorbent obtained in step (C), an elution solvent is added to the adsorbent and adjusted to basic, preferably pH 9-12, Alternatively, it is eluted by heating or stirring as appropriate, and the adsorbent is removed by a usual method such as filtration or centrifugation. As an elution solvent to be used, a basic solvent, for example, water adjusted to a basic pH, methanol, ethanol, isopropanol or the like, or an appropriate mixed solution thereof can be used, preferably water adjusted to pH 9 to 12. Can be used. The amount of the elution solvent can be appropriately set. In order to use the eluate thus obtained as a drug substance, the pH is appropriately adjusted to near neutral, etc. to finally obtain a vaccinia virus-inoculated rabbit inflammation skin extract (this extract). Can do.

  Since this extract is liquid at the time of completion, it can be made to have a desired concentration by appropriately concentrating and diluting. When manufacturing a formulation from this extract, it is preferable to heat-sterilize. In order to obtain an injection, for example, sodium chloride or the like can be added to prepare a solution that is isotonic with physiological saline. Moreover, oral solid preparations, such as a tablet, can also be manufactured by performing operation, such as concentration and drying suitable for this extract in a liquid state. Specific methods for producing such an oral solid preparation from this extract are described in the specifications of Japanese Patent Nos. 3818657 and 4883798. This preparation is an injection or a solid preparation for oral use thus obtained.

Examples of methods for administering a pharmaceutically effective amount to a patient in need of treatment include subcutaneous, intramuscular and intravenous administration in addition to oral administration, and the dosage depends on the type of inflammatory tissue extract inoculated with vaccinia virus. It can be set appropriately. The dose allowed for commercially available formulations is basically indicated as a medicinal product to administer 16 NU per day for internal use and 3.6 to 7.2 NU per day for injections. It can be appropriately increased or decreased depending on the degree of serious injury, patient differences, administration method, administration period, etc. (NU: neurotropin unit. Neurotropin unit is a SART stress mouse, which is a chronic stress animal whose pain threshold is lower than that of normal animals, Tested according to the modified Randall-Selitto method and defined by the ED ₅₀ value of analgesic potency, 1NU is the activity showing 1 mg of the analgesic activity-containing component of the neurotropin preparation when the ED ₅₀ value is 100 mg / kg. )

  Examples of the method for producing the extract and the results of pharmacological tests on the novel pharmacological action of the extract and patella cartilage softening are shown below, but the present invention is not limited by the description of these examples. Absent.

Example 1 (Production of the present extract)
The skin of healthy mature rabbits was inoculated intradermally with vaccinia virus, and the cut skin was cut out and collected. The collected skin is washed and disinfected with a phenol solution, then the excess phenol solution is removed, crushed, the phenol solution is added and mixed, left to stand for 3-7 days, and then stirred for another 3-4 days Warmed to 35-40 ° C. Thereafter, the extract obtained by solid-liquid separation was adjusted to pH 4.5 to 5.2 with hydrochloric acid, heat-treated at 90 to 100 ° C. for 30 minutes, and then filtered to deproteinize. Further, the filtrate was adjusted to pH 9.0 to 9.5 with sodium hydroxide, heated at 90 to 100 ° C. for 15 minutes, and then separated into solid and liquid.

  The obtained deproteinized solution was adjusted to pH 4.0 to 4.3 with hydrochloric acid, activated carbon in an amount of 2% of the deproteinized solution mass was added and stirred for 2 hours, followed by solid-liquid separation. Water was added to the collected activated carbon, adjusted to pH 9.5 to 10 with sodium hydroxide, stirred at 60 ° C. for 90 to 100 minutes, and then centrifuged to obtain a supernatant. Water was added again to the activated carbon precipitated by centrifugation, and then adjusted to pH 10.5 to 11 with sodium hydroxide, stirred at 60 ° C. for 90 to 100 minutes, and then centrifuged to obtain a supernatant. Both supernatants were combined and neutralized with hydrochloric acid to obtain the present extract.

Example 2 (Test method)
Next, an example of the test result about the effect | action (improvement or therapeutic effect) with respect to the patella cartilage softening of the formulation (this formulation) containing this extract obtained in the said Example 1 as an active ingredient is shown.

  In order to examine the effect of this preparation on patella cartilage softening, a test was conducted in humans as follows.

(1) Exam (Exam period: January 2011 to September 2012)
Thirty 60 patients diagnosed with patella cartilage softening were randomly divided into two groups, the preparation-administered group and the control group. To the control group, 2.0 mL of hyaluronic acid was administered once weekly and 5 times by intra-articular injection. For this preparation administration group, 2.0 mL of hyaluronic acid and one tube of “neurotropin injection solution 3.6 units” (trade name of injection in this preparation) were administered once weekly and five times by intra-articular injection. For intra-articular injection, the outer puncture method of the patellofemoral joint was used. In a supine position, bend the patient's knee joint 15 to 20 degrees, disinfect the skin in the usual way, and use a 5 mL syringe with the puncture site between 1/3 of the outer edge of the patella and the lateral condyle of the femur After inserting into the knee joint sac along the patellofemoral joint cavity and pulling the syringe to confirm that there was joint fluid, suck the joint fluid as completely as possible and leave it in the joint and gradually inject into the joint did. Basically, no other drugs were used. The therapeutic effect was determined after the treatment was completed.

(2) Determination of therapeutic effect The following diagnostic criteria were used to determine the therapeutic effect.
“Healing” was a complete recovery.
“Remarkable effect” was defined as a state in which main symptoms such as joint pain and swelling disappeared, joint function almost recovered, and normal activity was possible.
“Effective” means that the main symptoms such as joint pain and swelling are almost disappeared, the joint function is almost recovered or clearly improved, the independence of daily life is improved, and the independent life can be almost completed, The ability to work was partially recovered.
“Ineffective” was defined as a state in which the main symptoms such as joint pain and swelling were not improved, and the function of the joint activity was not improved compared to before treatment.
The therapeutic effect was determined using the above diagnostic criteria, and the cure rate, remarkable efficiency, effective rate, and total effective rate for all cases in each group were calculated. The results are shown in Table 1.
The two groups were evaluated by the knee joint Lysholm score before and 1 month after treatment. The results are shown in Table 2.

(3) Statistical analysis The determination of the therapeutic effect between the two groups was tested using the chi-square test. Paired t-tests were used for Lysholm scores before and after treatment in the same group, and independent sample t-tests were used for Lysholm scores before and after treatment in the two groups. In any test, the case where p <0.05 was regarded as “significant”.

  The test results in Table 1 and Table 2 were statistically analyzed. As a result, in the group in which this preparation was administered to patients with patella cartilage softening, the overall effective rate was significantly improved compared to the control group. From this, it was shown that this preparation is a drug having an effect of generally improving or treating the symptoms of patella cartilage softening.

  As described above, this preparation was found to have an excellent improvement or therapeutic effect on the symptoms of patella cartilage softening. In addition, this preparation has been used for many years and is recognized as a very safe drug. Therefore, this preparation is extremely useful as an agent for improving or treating patella cartilage softening.

Claims (12)

  An agent for improving or treating patella cartilage softening comprising an inflammatory tissue inoculated with vaccinia virus as an active ingredient.   The improvement or treatment agent according to claim 1, which is used in combination with a hyaluronic acid preparation.   The improvement or treatment agent according to claim 1 or 2 for knee pain due to patella cartilage softening.   The improvement or treatment agent according to any one of claims 1 to 3, wherein the inflamed tissue is an inflamed rabbit skin tissue.   The improvement or treatment agent according to any one of claims 1 to 4, which is an injection.   The improvement or treatment agent according to any one of claims 1 to 4, which is an oral agent.   Use of an extract of inflamed tissue inoculated with vaccinia virus for improvement of patella cartilage softening or manufacture of a therapeutic agent.   The use according to claim 7, wherein the ameliorating or treating patella cartilage softening is used in combination with a hyaluronic acid preparation.   The use according to claim 7 or 8 for improvement of knee pain due to patella cartilage softening or manufacture of a therapeutic agent.   The use according to any one of claims 7 to 9, wherein the inflamed tissue is a rabbit inflamed skin tissue.   The use according to any one of claims 7 to 10, wherein the ameliorating or treating agent for patella cartilage softening is an injection.   The use according to any one of claims 7 to 10, wherein the ameliorating or treating agent for patella cartilage softening is an oral agent.