Treatment for spinal cord injury

Abstract

[Problem] To provide a novel therapeutic agent containing a brilliant blue-based dye as an active ingredient. [Solution] This therapeutic agent for spinal cord injury contains, as an active ingredient, a brilliant blue-based dye (in particular, brilliant blue G), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

Description

[Technical Field]

[0001] The present invention relates to a therapeutic agent for spinal cord injury. [Background technology]

[0002] Republished Patent Publication No. 2012 / 066994 describes that Brilliant Blue G (BBG) has neuroprotective effects. U.S. Patent Publication No. 2010-74884 describes that Brilliant Blue G (BBG) is an antagonist (inhibitor) of P2X receptors and inhibits or blocks the pathological effects of ATP. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Re-tabled publication 2012 / 066994 [Patent Document 2] U.S. Patent Application Publication No. 2010-74884 Summary of the Invention [Problem to be solved by the invention]

[0004] The object of this invention is to provide a new therapeutic agent containing a brilliant blue dye as an active ingredient. [Means for solving the problem]

[0005] This invention is based on the finding from examples that administering brilliant blue dyes (particularly brilliant blue FCF and brilliant blue G) to a subject can treat spinal cord injury in the subject.

[0006] This invention relates to a therapeutic agent for spinal cord injury. The therapeutic agent for spinal cord injury means a pharmaceutical or veterinary drug for treating spinal cord injury in a patient (human or non-human mammal) suffering from spinal cord injury.

[0007] This therapeutic agent for spinal cord injury contains, as an active ingredient, a brilliant blue dye, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. Preferred examples of brilliant blue dyes are brilliant blue FCF and brilliant blue G, and of these, brilliant blue G is most preferred.

[0008] This therapeutic agent may be a therapeutic agent for herniated disc containing as an active ingredient either or both of a P2X7 inhibitor and a pannexin 1 inhibitor. As shown in the examples, brilliant blue FCF and brilliant blue G are highly effective in treating herniated discs, and therefore, it is believed that P2X7 inhibitors and pannexin 1 inhibitors are highly effective in treating herniated discs.

[0009] An example of this therapeutic agent for spinal cord injury is a therapeutic agent for herniated disc. The subject to which this therapeutic agent for spinal cord injury is administered may be a human or a mammal other than a human. Examples of mammals other than a human include dogs and cats. [Effects of the Invention]

[0010] According to this invention, it is possible to provide a new therapeutic agent (therapeutic agent for spinal cord injury) containing a brilliant blue dye as an active ingredient. [Brief explanation of the drawings]

[0011] [Figure 1] Figure 1 is a photograph in place of a drawing showing an MRI image of a dog with a herniated disc before treatment. [Figure 2] Figure 2 is a photograph, in place of a drawing, showing an MRI image of a dog with a herniated disc taken one month after administration of Brilliant Blue G. DETAILED DESCRIPTION OF THE INVENTION

[0012] The following describes embodiments for carrying out the present invention. The present invention is not limited to the embodiments described below, and also includes appropriate modifications of the embodiments below within the scope obvious to those skilled in the art.

[0013] This invention relates to a therapeutic agent for spinal cord injury. The therapeutic agent for spinal cord injury refers to a pharmaceutical or veterinary drug for treating spinal cord injury in patients (humans or non-human mammals) suffering from spinal cord injury. Examples of spinal cord injury include difficulty walking due to spinal cord injury, difficulty standing upright due to spinal cord injury, paralysis due to spinal cord injury, retrograde paralysis due to spinal cord injury, retrograde complete paralysis due to spinal cord injury, herniated disc, and spinal cord injury.

[0014] This therapeutic agent for spinal cord injury contains an effective amount of a brilliant blue dye, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient.

[0015] Brilliant blue dyes are known dyes, as described in, for example, Japanese Patent Application Laid-Open No. 2013-241431. Examples of brilliant blue dyes are brilliant blue FCF, brilliant blue G, and brilliant blue R. Preferred examples of brilliant blue dyes are brilliant blue FCF or brilliant blue G, with brilliant blue G being the most preferred. Brilliant blue G is a P2X7 antagonist, and brilliant blue FCF is a pannexin channel 1 antagonist. Thus, brilliant blue FCF and brilliant blue G have different functions. Of these, brilliant blue FCF is also preferred for the treatment of spinal cord injury, as shown in the examples. Brilliant blue FCF is a dye known as blue No. 1 and has excellent biocompatibility.

[0016] Examples of pharmacologically acceptable salts include salts formed with inorganic bases, ammonia, organic bases, inorganic acids, organic acids, basic organic acids, halide ions, etc., and internal salts. Examples of inorganic bases include alkali metals (Na, K, etc.) and alkaline earth metals (Ca, Mg, etc.). Examples of organic bases include trimethylamine, triethylamine, choline, procaine, ethanolamine, etc. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, etc. Examples of organic acids include p-toluenesulfonic acid, methanesulfonic acid, formic acid, trifluoroacetic acid, and maleic acid, etc. Examples of basic amino acids include lysine, arginine, ornithine, histidine, etc.

[0017] An example of a pharmaceutically acceptable solvate is a hydrate. An example of a hydrate is a monohydrate.

[0018] An example of this therapeutic agent for spinal cord injury is a therapeutic agent for herniated disc. The subject to which this therapeutic agent for spinal cord injury is administered may be a human or a mammal other than a human. Examples of mammals other than a human include dogs and cats.

[0019] The pharmaceutical preparation of the present invention preferably contains a brilliant blue dye at a concentration of 0.01-100 mg / ml, preferably 0.5-50 mg / ml, and most preferably 0.1-20 mg / ml. The dosage of the brilliant blue dye can be adjusted appropriately depending on the subject's weight, type, and age. Examples of dosages based on body weight include 0.1 mg / kg / day to 1 g / kg / day, 1 mg / kg / day to 100 mg / kg / day, 2 mg / kg / day to 50 mg / kg / day, and 5 mg / kg / day to 20 mg / kg / day.

[0020] The drug of the present invention may be a therapeutic agent for herniated discs containing either or both of a P2X7 inhibitor and a pannexin 1 inhibitor as active ingredients. As shown in the examples, Brilliant Blue FCF and Brilliant Blue G are highly effective in treating herniated discs, and therefore P2X7 inhibitors and pannexin 1 inhibitors are considered to be highly effective in treating herniated discs. These active ingredients may be appropriately adjusted by incorporating an effective amount into a pharmaceutical or veterinary drug, similar to those containing a brilliant blue dye as an active ingredient. The effective amount may be approximately the same as that of the above-mentioned brilliant blue dye.

[0021] A P2X7 inhibitor refers to a substance that exhibits receptor affinity for the P2X7 receptor and inhibits the function of the receptor. P2X7 inhibitors are known. Examples of P2X7 inhibitors include KN62 (4-[(2S)-2-[(isoquinolin-5-ylsulfonyl)(methyl)amino)-3-oxo-3-(4-phenylpiperazin-1-yl)propyl]phenyl isoquinoline-5-sulfonate), MRS2306, A-759020, AZD9056, and CE-224535, as described in WO2012036193A1.

[0022] Examples of pannexin 1 inhibitors are carbenoxolone, probenecid, and pharmaceutically acceptable salts thereof.

[0023] The dosage form of the drug of the present invention is not particularly limited. An example of the dosage form is an injection. To produce an injection, the active ingredient is mixed with a known carrier (water, etc.) and sealed in a vial, syringe, or ampoule. Known methods can be used as appropriate to produce the dosage form.

[0024] The therapeutic agent for spinal cord injury of the present invention may be administered orally. In this case, it can be formulated by mixing the active ingredient, brilliant blue dye, and excipients with a known carrier and tableting the mixture with a tableting machine.

[0025] This specification also provides a method for treating spinal cord injury, comprising the step of administering to a subject a brilliant blue-based dye, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

[0026] Manufacturing Example 1 Brilliant Blue G (1 g) was dissolved in an appropriate amount of water for injection, and water for injection was added to make 100 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating.

[0027] Manufacturing Example 2 An appropriate amount of saline was prepared, 1 g of brilliant blue G was dissolved in it, and saline was added to make 100 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating.

[0028] Manufacturing Example 3 An appropriate amount of saline was prepared, 1 g of brilliant blue G was dissolved in it, and saline was added to make up to 1000 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating.

[0029] Production Example 4 Brilliant Blue FCF (1 g) was dissolved in an appropriate amount of water for injection, and water for injection was added to make up to 100 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating.

[0030] Manufacturing Example 5 An appropriate amount of saline was prepared, 1 g of Brilliant Blue FCF was dissolved in it, and saline was added to make up to 100 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating.

[0031] Manufacturing Example 6 An appropriate amount of saline was prepared, 1 g of Brilliant Blue FCF was dissolved in it, and saline was added to make 1000 mL. After filtering, the solution was filled into glass vials, sealed, and sterilized by heating. [Example]

[0032] Treatment of a dog with intervertebral disc herniation accompanied by delayed complete paralysis using Brilliant Blue G or Brilliant Blue FCF

[0033] Prescription and Results Brilliant Blue (FCF or G) was administered intravenously at a dose of 10 mg / kg / day for one week (orally in one case) and patients were kept at rest in the hospital. None of the patients underwent any surgical procedures such as laminectomy, and after one week of medication, they were maintained at home. It has currently been prescribed to over 20 cases (prescribed only for those with complete paralysis (grade 4-5)), and so far, all cases (100%) have shown improvement (becoming able to walk) within two weeks to one and a half months after treatment.

[0034] Case 1 (first case in 2013) Miniature Dachshund (dog), 10 years old, male The cat suddenly barked the night before and then came to the hospital with complete paralysis (loss of deep pain sensation, no movement of the tail, no movement of the hind legs. Walks dragging with only the front legs. Bladder paralysis also occurs, and urination is achieved by pressure).

[0035] prescription Brilliant Blue FCF was administered intravenously at a dose of 10 mg / kg / day for one week. No decompression surgery such as hemilaminectomy was performed.

[0036] Progress At the end of the treatment (1 week later), the patient was still in a state of retrograde paralysis. Two weeks after the start of treatment, the patient's tail began to move and deep pain sensation returned to some extent. Three weeks later, the patient was able to stand up. After that, he was able to walk independently within a month and a half, and thereafter the paralysis improved to a normal level, and even after 7 years (he was over 17 years old; the average lifespan of a dog is 14-15 years), there was no recurrence and he was able to walk normally.

[0037] Case 2: Miniature Dachshund (dog), 6 years old, male Transferred from another hospital due to complete paralysis (grade 5) (surgery was required at the university hospital) The prescription was the same as above (Brilliant Blue FCF was administered intravenously at a dose of 10 mg / kg / day for one week, without any decompression surgery such as hemilaminectomy).

[0038] Progress After 2 weeks, the patient was able to walk normally.

[0039] Case 3: Maltese dog, 8 years old, male Transferred from another hospital due to complete after-effect paralysis (grade 5) The prescription was Brilliant Blue FCF administered intravenously at a dose of 10 mg / kg / day for one week, without any decompression surgery such as hemilaminectomy.

[0040] Progress After 2 weeks, the patient was able to walk normally.

[0041] Case 4: Wirehaired Dachshund (dog), 10 years old, female The day before, he suddenly started crying loudly, then developed complete paralysis (grade 5) and came to the hospital. The prescription was the same as above (Brilliant Blue FCF was administered intravenously at a dose of 10 mg / kg / day for one week, without any decompression surgery such as hemilaminectomy).

[0042] Progress After 2 weeks, the patient was able to walk normally.

[0043] Case 5: Mixed breed (Kai dog), 18 years old, female The patient had been experiencing dizziness (post-stroke paresis) for the past week and had been unable to stand since the previous day, and visited the hospital (grade 5). The prescription was the same as above (Brilliant Blue FCF was administered intravenously at a dose of 10 mg / kg / day for one week, without any decompression surgery such as hemilaminectomy).

[0044] Progress After one week, he was able to stand with support, and after two weeks he was able to walk independently.

[0045] Case 6: Miniature Dachshund (dog), 14 years old, male The day before, he suddenly started crying loudly and developed complete paralysis (grade 5) and came to the hospital. Prescription: Brilliant Blue G was administered intravenously at a dose of 10 mg / kg / day for one week. No decompression surgery such as hemilaminectomy was performed.

[0046] Progress After 7 weeks, the patient was able to walk, although he still had some incomplete paralysis, and by 9 weeks he was able to walk almost normally, although he was still slightly unsteady.

[0047] Case 7: Miniature Dachshund (dog), 14 years old, female The patient presented with retrograde paralysis (grade 4) and was unable to walk despite experiencing deep pain sensations. prescription Brilliant Blue FCF was orally administered at a dose of 20 mg / kg / day in the morning and evening for one week. No decompression surgery such as hemilaminectomy was performed.

[0048] Progress Two weeks later, although he was still partially paralyzed, he was able to walk and has been able to do so ever since.

[0049] Other cases without video data This prescription has been administered to over a dozen miniature schnauzers, miniature dachshunds, chihuahuas, and other dogs (completely paralyzed or nearly so: at least unable to walk), and all of them (100%) have been able to walk.

[0050] Case 8: Miniature Dachshund (dog), 14 years old, male, grade 5 (complete paralysis) Brilliant Blue G was administered intravenously at a dose of 10 mg / kg / day for one week. After six weeks, the patient was able to walk. There has been no recurrence for over one year.

[0051] Case 9: Miniature Dachshund (dog), 13 years old, female, Grade 4 (Although there was some deep pain, the dog was unable to walk or stand up for three weeks. The patient was transferred from another hospital and also had bladder paralysis.) Brilliant Blue G was administered intravenously at a dose of 10 mg / kg / day for one week. After two weeks, the patient was able to walk. There has been no recurrence for over five months.

[0052] Case 10: Miniature Dachshund (dog), 8 years old, female, Grade 5 (no deep pain sensation, unable to walk, unable to stand, transferred from another hospital for one week) Brilliant Blue G was administered intravenously at a dose of 10 mg / kg / day for one week. The patient was able to walk after two weeks. As shown in Figure 1, a pre-treatment MRI image showed approximately 50-60% compression of the spinal nerves. After two weeks of administration, the patient was able to walk. A repeat MRI was performed one month later, as shown in Figure 2. The degree of spinal cord compression remained largely unchanged, but the patient was able to run around at that point, and the grade assessment had improved from Grade 5 to Grade 0. This is the first data available anywhere in the world, suggesting that the degree of intervertebral disc compression does not affect gait improvement. This is at least the first finding in the world to demonstrate that walking is possible without the decompression treatment typically performed through disc herniation surgery. [Example]

[0053] Report on the use of Brilliant Blue for cats Japanese cat, age unknown (estimated age: 4-5 years old), male Visited the hospital after being unable to move due to a traffic accident The animal was lying on its side, unable to move, with constricted pupils and no consciousness. It had a skull fracture and fractures at T13, L1 and L2. Its hindquarters were paralyzed and it had no deep pain sensation. The chest bruises were in extremely bad condition, and although euthanasia was considered, treatment was attempted.

[0054] Symptomatic treatment: Dexamethasone, Stadol, Victas, Vitamin B1, Sollact drip infusion Brilliant Blue 10 mg / kg / day intravenous infusion Dissolve 0.7g (700mg) of Brilliant Blue in 14ml of Sollact (infusion solution) to make 50mg per ml of medication. For a cat weighing 3kg, infuse 0.6ml of 30mg intravenously per day.

[0055] The next day, his eyes opened, his pupils disappeared, and he was able to lie down and lift his head a little.

[0056] Consideration It is estimated that there are 100,000 people nationwide who are wheelchair-bound or bedridden due to spinal cord injuries. Because the central nervous system generally does not regenerate after injury, spinal cord injury, regardless of species, significantly reduces quality of life (QOL) due to inability to stand and urinary and bowel problems. Approximately 1% of the human population (approximately 1.26 million people in Japan) suffers from lumbar disc herniation. Because pain and numbness reduce QOL and medical treatment is unlikely to result in a complete cure, surgical treatment is often the preferred option. It is estimated that there are more than 100,000 dogs with herniated discs in Japan. Many dogs with complete paralysis (grade 4 or higher) who are unable to walk opt for surgery, but the cost is high, costing hundreds of thousands of yen, and involves risks such as anesthesia, and the success rate is low. In particular, many dogs with complete paralysis (grade 5), who have lost their deep pain sensation, do not respond to surgical treatment, so surgery is not an option. In this situation, our treatment with Brilliant Blue G improved the paralysis in all (100%) of the Grade 5 dogs, for which no other treatment was available, and enabled them to walk. Furthermore, as shown in Figures 1 and 2, a comparison of MRI images taken before and after administration of Brilliant Blue G revealed that the intervertebral disc material remained compressed on the spinal nerves, with no change. However, because the dogs had improved to the point where they no longer appeared to feel pain and were able to run around, surgery to remove the intervertebral disc material, such as hemilaminectomy, which is routinely performed in both human and veterinary medicine, is deemed unnecessary.

[0057] When the spinal cord is damaged by a herniated disc or other cause, large amounts of adenosine triphosphate (ATP) are released from the damaged cells. The released ATP binds to and activates the P2X7 receptors of immune cells (especially neutrophils, which are the first to enter) and microglia, causing inflammation to spread to spinal nerves other than the physically damaged area. In addition, the infiltration of microglia into the damaged area causes scar formation, reducing the space for nerve cell regeneration, and therefore preventing nerve repair and reconnection.

[0058] Brilliant Blue FCF is thought to inhibit pannexin 1, while Brilliant Blue G inhibits the actions of pannexin 1 and P2X7. In other words, when the spinal cord is damaged by a herniated disc or other condition, Brilliant Blue FCF and Brilliant Blue G inhibit the inflammatory response (particularly neutrophil activity) caused by excessive ATP release. This suppresses secondary damage and scarring caused by microglial infiltration, allowing nerve cells to regenerate and improving paralysis caused by spinal cord injuries, including herniated discs.

[0059] Pannexin 1 inhibitors such as carbenoxolone and P2X7 inhibitors such as KN62 may also be effective in treating herniated discs, similar to Brilliant Blue FCF and Brilliant Blue G. For many years, surgery was considered the only curative treatment for herniated discs in both human and veterinary medicine. However, with Brilliant Blue FCF or Brilliant Blue G, complete recovery through medical treatment is now possible. We believe that Brilliant Blue FCF or Brilliant Blue G will certainly brighten and enrich the lives of people and animals who have suffered from herniated discs and a decline in their quality of life. [Industrial Applicability]

[0060] The present invention can be used in the fields of pharmaceuticals and veterinary medicines.

Claims

1. A therapeutic agent for dogs suffering from hindquarters incomplete paralysis or complete hindquarters paralysis due to intervertebral disc herniation, the therapeutic agent for dogs containing, as an active ingredient, a brilliant blue dye, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, wherein the brilliant blue dye is brilliant blue FCF or brilliant blue G.

2. A therapeutic agent for dogs that have been suffering from complete hindquarters paralysis due to intervertebral disc herniation for a period of one to three weeks, the therapeutic agent containing as an active ingredient a brilliant blue dye, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, wherein the brilliant blue dye is brilliant blue FCF or brilliant blue G.

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