WO2006082771A1 - Preventive or therapeutic agent for stroke or sequelae of stroke comprising as main component 1-aminocyclopropanecarboxylic acid or the like - Google Patents

Abstract

[PROBLEMS] To provide a novel preventive or therapeutic agent for stroke or sequelae of stroke. [MEANS FOR SOLVING PROBLEMS] A preventive or therapeutic agent for stroke or sequelae of stroke comprising as a main component, a compound selected from 1-aminocyclopropanecarboxylic acid, its prodrugs, and their pharmaceutically acceptable salts and hydrates has been developed. For example, by being administered at 50 mg/kg, which is one-hundredth or less of the acute toxic level, 1-aminocyclopropanecarboxylate hydrochloride monohydrate reduces the occurrence of stroke by 90% and even in the case where stroke occurs, the sequelae are mild.

Description

 Specification

 Drugs for the prevention or treatment of stroke or sequelae of stroke with 1-aminocyclopropanecarboxylic acid as the main component

 Technical field

 [0001] The present invention relates to a novel agent for preventing or treating stroke or stroke sequelae.

 Background art

 [0002] According to statistics from the Ministry of Health, Labor and Welfare in 2001, the number one cause of the need for nursing care for 94,75 0 elderly people aged 65 years or older is the cerebrovascular disease such as stroke. There are 24,749 people, 2nd place is 11,744 fractures, 3rd place is dementia 10,659 people, stroke and other cerebrovascular diseases are overwhelming. Development of drugs for preventing or treating stroke or sequelae of stroke is strongly desired. Elderly hypertensive patients, diabetic patients, and hyperlipidemia patients are prone to stroke. Therefore, as a prevention of stroke or stroke sequelae, a method of managing hypertension that causes stroke or stroke sequelae with antihypertensive agents, diabetes with antidiabetic agents, and hyperlipidemia with antilipidemic agents is adopted. However, it is sufficient to prevent the occurrence of stroke or stroke sequelae. Accordingly, research and development of drugs for preventing or treating stroke or sequelae of stroke are being conducted in various approaches. For example, treatment of thrombotic symptoms in atherosclerotic stroke with thromboxane A2 synthesis inhibitor (Patent Document 1), prevention of recurrence of stroke at a dose that does not lower blood pressure (Patent Document 2), NR2B subtype selective NMDA reception Stroke treatment combining somatic antagonists with various drugs (Patent Document 3), Stroke preventive agent containing phosphatidylcholine as an active ingredient (Patent Document 4), Peroxidation containing phofen as an active ingredient is reduced Preventive stroke (patent document 5), stroke preventive agent containing xanthine as an active ingredient (patent document 6), stroke preventive agent by improving hypertension symptoms containing dioxanecyclo [3, 3, 0] octane derivative as an active ingredient ( Patent literature 7) etc. are published. However, the development of new drugs for the prevention or treatment of new stroke or sequelae of stroke, which is still insufficient, is desired.

On the other hand, 1-aminocyclopropane carboxylic acid, also known as alpha-aminocyclopropane carbo Acid (English name: l-aminocyclopropanecarboxylic 'acid; hereinafter abbreviated as ACC;;) is a natural substance extracted from concentrated fruit juice (Non-Patent Document 1) and is an odorless, water-soluble white crystalline substance. A type of growth regulator that has one amino group, one carboxylic acid, and one three-membered ring. ACC can be obtained from synthetic methods (Patent Document 8, Non-Patent Documents 2-4) as well as plant extraction methods.

 ACC is a plant growth-regulating action, ethanol consumption action (Non-patent Document 5), as well as the strykinin-insensitive glycine receptor site of N-methyl-D-aspartate (hereinafter abbreviated as NMDA) receptor complex. A non-patent document (Non-patent document 6) and an antagonist of the glutamate receptor site (Non-patent document 7). ACC suppresses degeneration of hippocampal nerves induced by NMDA (Non-patent Document 8), protects spinal nerves against NMDA toxicity (Non-patent Document 9), and improves memory through NMDA receptors (Patent Document) 9) The ability to prevent neuropathy caused by focal ischemia, extensive ischemia, and spinal cord ischemia Long-term administration of ACC reduces the expression of NMDA receptor messenger RNA and reduces the efficacy (Non-patent Document 10). Thus, it appears that ACC acts on NMDA receptors to protect but not sustain nerves and does not prevent or treat stroke or stroke sequelae.

 In the following, prior documents are displayed separately for patent documents and non-patent documents.

Patent Document 1: Japanese Patent Laid-Open No. 2003-26660

Patent Document 2: Japanese Patent Laid-Open No. 2002-370981

Patent Document 3: Japanese Patent Laid-Open No. 2002-332095

Patent Document 4: Japanese Unexamined Patent Publication No. 2000-239168

Patent Document 5: Japanese Patent Laid-Open No. 11-228396

Patent Document 6: Japanese Patent Laid-Open No. 10-182469

Patent Document 7: JP-A-8-268887

Patent Document 8: Japanese Patent Laid-Open No. 7-278077

Patent Document 9: JP-A-8-510990

Non-Patent Document 1: F. Burroughs, Nature, 1957 179 卷 360

Non-Patent Document 2: A. Connors et al., J. Chem. Soc. 1960, page 2129

Non-Patent Document 3: E. Bunuel et al., Synlett. 1992 7-7 579 Non-Patent Document 4: X. Zhu et al., Synthetic Communication 1998 28-17, 3159 Non-patent document 5: BA McMillen et al., Brain-Res. Bull. 2004 64-3, 279 Non-patent document 6: R. Trullas et al., Pharmacol. Biochem. Behav. 1989 34-2 31

3 pages

 Non-Patent Document 7: R. Nahum-Levy et al., Molecular Pharmacology 1989 56 卷 12 07

 Non-Patent Document 8: A. Zapata et al., Neuroreport. 1996 7-2 397

 Non-Patent Document 9: Y. Lin et al., Eur. J. Pharmacol. 1996 318-2 2-3 pages 491 Non-Patent Document 10: S. Bovetto et al., Am. Soc. For-Pharmacol. Exp. Therap. 1997 293 卷 3 1503

 Disclosure of the invention

 Problems to be solved by the invention

[0005] An object of the present invention is to provide a new agent for preventing or treating stroke or sequelae of stroke.

 Means for solving the problem

 [0006] In view of this, the inventors of the present invention compared the effects of ACC on the antihypertensive effect of blood pressure, the incidence of stroke, the severity of sequelae of stroke, major organs, and lifespan in comparison with control rats using ACC. As a result of confirming that ACC has a significant effect on the prevention or treatment of stroke or stroke sequelae in stroke-prone rats SHRSP, the present invention was completed as a result of various further studies based on this result.

 [0007] That is, the present invention is directed to "ACC and prodrugs thereof, and pharmaceutically acceptable salts thereof, in response to the problem of" providing new drugs for preventing or treating stroke or sequelae of stroke ". And a pharmacological agent for preventing or treating stroke or sequelae of stroke, comprising a selected compound as a main component.

 The invention's effect

[0008] The drug of the present invention is useful as a drug for preventing or treating stroke or stroke sequelae. BEST MODE FOR CARRYING OUT THE INVENTION

 [0009] The present invention is a novel drug for preventing or treating stroke or sequelae of stroke. The main components of the drug of the present invention are ACC and prodrugs thereof, and pharmaceutically acceptable salts and hydrate powers thereof.

 [0010] The ACC can be a natural substance or a synthetic product. Natural ACC is contained in pears, apple juice, etc. Power ACC obtained by extraction from pear apple juice concentrated by the method described in Non-Patent Document 1 mentioned above is found in various other fruits. Because it is included, it can also be extracted from other fruits. As the synthetic ACC, the force obtained by the method described in Non-Patent Documents 2 to 4 described above, and those synthesized by other methods can be used. Among various synthesis methods, the method described in Non-Patent Document 4 can synthesize ACC in a high yield under mild reaction conditions using easily obtainable materials. desirable. For example, cyclopropane-1,1-dicarboxylic acid (hereinafter abbreviated as Compound 1) is obtained from jetylmalonic acid and 1,2-dibromoethane. Acetic anhydride, concentrated sulfuric acid and acetone are added to compound 1 to obtain spiro- [2,5] -5,7-dioxa-6,6-dimethyloctane-4,8 dione (hereinafter abbreviated as compound 2). . Compound 2 is decomposed with hydrazine to give 1-hydrazinocarbocyclopropane-1-carboxylic acid (hereinafter abbreviated as Compound 3). ACC is obtained by the Curtius rearrangement of 1-azidocarbonylcyclopropane-1-force rubonic acid (hereinafter abbreviated as Compound 4) produced by treating Compound 3 with hydrochloric acid followed by sodium nitrate. Many companies sell ACC. Commercially available ACC can also be used. As shown in the examples, ACC has acute toxicity LD50 6.81 g Z kilogram, body weight, food utilization rate, blood image, biochemistry, organs and other subacute toxicity especially bone marrow micronucleus rate, malformation rate, mutagen, There is no toxicity that can be administered to humans without special toxicity that affects reproduction, etc.

 [0011] The prodrug of ACC may be any as long as it is pharmaceutically acceptable as long as it is converted to ACC by hydrolysis or the like. Examples of such prodrugs include, but are not limited to, methyl ester derivatives or ethyl ester derivatives at the carboxyl group in the ACC molecule, and acetyl derivatives at the amino group in the ACC molecule. It may be synthesized in the future.

[0012] ACC salt and ACC prodrug salt are limited to pharmaceutically acceptable salts. It is. Examples of pharmaceutically acceptable salts include alkali metal salts such as sodium and potassium, alkaline earth metal salts such as magnesium and calcium, and ammonia such as ammonium and tetramethylammonium. Salts, organic acid salts such as triethylamine, lysine and arginine, and acid addition salts with mineral acids such as hydrochloric acid, hydrobromic acid and sulfuric acid, but are not limited thereto. Any salt that can be synthesized in the future may be used as long as it is pharmaceutically acceptable as long as it is pharmaceutically acceptable.

 [0013] Hydrates of ACC and prodrugs thereof, pharmaceutically acceptable salts of ACC and prodrug salts thereof can also be used as the main component of the present invention. ACC hydrates include ACC (H20), ACC (H20) 2, ACC (H20) 3, ACC (H20) 4, AC C (H20) 5, ACC (H20) 6, ACC (H20) 7, ACC (H20) 8 is mentioned. Examples of hydrates of ACC salts include ACC · HC1 (H20).

[0014] Since the main component of the medicament for preventing or treating stroke or sequelae of stroke of the present invention is a non-high molecular weight molecule, it is administered orally or parenterally (suppository, intramuscular, subcutaneous, intravenous, intracerebral, etc. ) Can be administered. Even if the main component is a prodrug of ACC, the prodrug is activated in the body, so it is not necessary to activate the prodrug in advance.

 [0015] When preparing an oral preparation, after adding an excipient and, if necessary, an antioxidant, a binder, a disintegrant, a lubricant, a coloring agent, a flavoring agent, etc., a conventional method is added. To form tablets, coated tablets, granules, capsules, solutions, syrups, elixirs, oily or aqueous suspensions. Examples of the excipient include lactose, corn starch, sucrose, glucose, sorbit, and crystal cell loss. Examples of the binder include polyvinyl alcohol, polyvinyl ether, ethyl cellulose, methinorescenellose, gum arabic, tragacanth, gelatin, shellac, hydroxypropyl cellulose, hydroxypropyl starch, and polyvinylpyrrolidone.

[0016] Examples of the disintegrant include starch, agar, ungelatinized, crystalline cellulose, calcium carbonate, sodium hydrogen carbonate, calcium citrate, dextran, and pectin. Examples of the lubricant include magnesium stearate, talc, polyethylene darcol, silica, hydrogenated vegetable oil, and the like. As the colorant, those permitted to be added to pharmaceuticals can be used. Examples of flavoring agents include cocoa powder, noh, katsu brain, Perfume acid, hearth oil, dragon brain, cinnamon powder, etc. can be used. These tablets may be appropriately coated on the granules by sugar coating, gelatin coating, etc. if necessary.

 [0017] When preparing an injection, if necessary, an isotonic solution such as glucose or physiological saline, a pH adjuster, a buffer, an antioxidant and other stabilizers, a preservative and the like are added. According to the method, the injection should be subcutaneous, intramuscular, intravenous and intracerebral. The injection may be a solid preparation or a preparation for daily use by freezing and drying after storing the solution in a container. One dose may be stored in a container, and multiple doses may be stored in the same container. In the case of a solution, it is better to shut off oxygen and moisture. Therefore, in some cases, it is better to put it in an ampoule type container and evacuate it or replace it with nitrogen gas. Any known method may be used.

 [0018] The dose of the agent for preventing or treating stroke or sequelae of the present invention varies depending on the dosage form such as oral preparation, suppository, subcutaneous injection, intramuscular injection, intravenous injection, intracerebral injection and the like. However, in the case of humans, adults are usually administered in daily doses of 0.01 to: LOOO milligrams, preferably 0.1 to 100 milligrams, and 1 dose is given in 1 to 1 doses or divided into 2 to 4 doses. . The dose may be adjusted according to age and symptoms.

 EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples. Regarding ACC toxicity and efficacy tests, acute toxicity test, subacute toxicity test, teratogenic toxicity test, mutagenicity test, white blood cell in reproductive test, phosphorus, neutrophil, hemoglobin, GOT, total serum Analyzes of protein, albumin, triglyceride, cholesterol, glucose, urea, micronucleus rate, sperm malformation rate, reverse mutant, dead fetal rate, fetal weight, height / tail length, sternum deletion rate, etc. Medical research and other toxicity test contracts are routinely commissioned and analyzed by urine N02ZN03, Creathun, Na, K, Ca, Cl, Mg, blood albumin, globulin, GOT, GTP, gamma-daltamyl transpeptidase, Urine nitrogen, Creathun, Na, K, Ca, Cl, Mg, angiotensin 1 convertase, angiotensin 2 etc. are analyzed by clinical laboratory contractors such as Shionogi 'Biomedical' Laboratories. It was commissioned analysis, Ru.

 Example 1

[0019] Calories 26 grams of Compound 1 in 24 milliliters of acetic anhydride in a bath of ice salt at 0 degrees Celsius While suspended by stirring, 0.8 ml of 98% concentrated sulfuric acid and 20 ml of acetone were added dropwise. The insoluble material melted and became a clear solution. This solution was further stirred for 1 hour in an ice-salt bath, placed in a refrigerator, and left for 1 hour. The next day, the solution was diluted with 300 milliliters of ice water. The resulting crystals were separated by filtration and washed twice with 300 ml of ice water to obtain the first crystals. The filtrate and the washing solution were combined, neutralized to pH 5 with saturated sodium bicarbonate, and the resulting crystals were separated by filtration and washed twice with a small amount of ice water to obtain a second crystal. The filtrate was extracted three times with 50 ml of ether, the ether layer was separated and evaporated to give a third crystal. 1st power Disliked crystals up to 3rd and dried, yielding 29 grams of crystals. From the melting point (61-63 degrees Celsius) and the RF value (0.34) of the thin-layer chromatograph developed with cyclohexane and acetone (3: 1 volume ratio), it was confirmed that this crystal was compound 2. .

 Example 2

 [0020] 17 grams of Compound 2 obtained in Example 1 was suspended in 100 ml of absolute ethanol at room temperature (21-25 degrees Celsius), and 10 grams of 85% hydrazine hydrate and anhydrous ethanol were suspended in the suspension. Knoll 10 ml of the mixed solution was added dropwise. The insoluble material melted and became a clear solution. This clear solution was left at room temperature for 4 hours. The disappearance of compound 2 in the solution was confirmed by the thin layer chromatograph described in Example 1. The solution was evaporated, and the resulting white needle crystals were filtered with a suction pump, and the crystals were washed with a small amount of absolute ethanol and dried. 14. Four drams of dry crystals were obtained. This crystal was confirmed to be Compound 3 from the melting point (150-152 degrees Celsius) and the R-value (0.5) of the thin-layer chromatograph developed with ethanol.

 Example 3

[0021] 5 grams of compound 3 obtained in Example 2 was dissolved in 10 milliliters of water at room temperature, and 5 milliliters of 36-38% concentrated hydrochloric acid was added to the solution, and 15 grams of crushed ice and It was poured into ether 20 ml and stirred to suspend insoluble matter. While stirring the suspension in an ice-salt bath, 5.7 ml of 34.5% aqueous sodium nitrite solution was slowly added drop by drop. The insoluble material melted and became a clear solution. This clear solution was stirred for an additional 20 minutes. During operation, the temperature of the mixture was maintained between 0 degrees Celsius and minus 5 degrees Celsius. The ether layer was separated. The aqueous layer was extracted twice with 10 ml of cooled ether. The resulting three ether solutions And dried for 15 minutes over anhydrous calcium chloride. The dried ether was transferred into 30 milliliters of benzene in a flask in a 40 degree Celsius bath over 1 hour. Ether and nitrogen in the benzene solution were evaporated at room temperature. 12 ml of 36-38% concentrated hydrochloric acid was added drop by drop to a benzene solution in which ether and nitrogen had been evaporated. Benzene and carbon dioxide were removed under reduced pressure. The remaining solution was diluted with water, adjusted to pH 1 with concentrated hydrochloric acid, recrystallized with ethanol and water, and dried at room temperature under reduced pressure. 2. 5 grams of dry crystals were obtained. R-value (0.42) of thin layer chromatograph developed with melting point (215-220 degrees Celsius), n-butanol, acetic acid and water (6: 2: 2 volume ratio), infrared spectrum IR (KBr) (30 50, 3050-2800, 2640-2440, 2100, 1650-1540, 1410, 1635 cm-1), proton nuclear magnetic resonance spectrum 1H-NMR (CF3C00D ゝ 79.542MHz) (1.83 (s, 2H), 1.95 ( s, 2H) ppm), mass spectrum MS (mZz, EI) 101 (M +), 83, 56, 55, 5 4, 38, 36, 28, elemental analysis (C31.06, H6.48, N8. 97 From C122. 79), it was confirmed that this crystalline force was SACC hydrochloride monohydrate C4H10C1NO3.

 Example 4

 [0022] 450 g of ACC hydrochloride monohydrate was synthesized in the same manner as in Examples 1 to 3. Using this, we conducted an acute toxicity test, a subacute toxicity test, a micronucleus test, a mutagenicity test, a reproductive effect test, etc., in accordance with the “Food Safety Toxicology Evaluation Procedure Ordering Method”.

 Example 5

 [0023] Twenty healthy males and females weighing 19 to 22 grams were divided into 5 groups per group and 4 groups of males and females. The ACC hydrochloride monohydrate obtained in Example 4 was 1.00, 2. 15, 4, 64, and 10.0 (gram Z kilogram body weight) were administered by oral gavage with a sonde (in order, 1.00 group, 2.15 group, 4.64 group, 10.0 group). We observed for 7 days and examined acute toxicity. In groups 1.00, 2.15, and 4.64, both male and female groups were deadly. In group 10.0, 5 of 5 animals died within 30 minutes after oral gavage. Calculations showed that LD50 was 6.81 grams / kilogram for both males and females.

 Example 6

[0024] 40 healthy male and female SD rats weighing 90-120 grams were divided into 10 groups per group and 4 groups of males and females, each day. 0, 0.125, 0.25, 0.50 (gram Z kilogram body weight) by oral gavage with a sonde for 30 days (each group in turn 0.0 group, Group 0, 125, 0.25, and 0.50. ) And observed for 30 days to examine subacute toxicity. The weights of the 0.0, 0.125, 0.25, and 0.50 groups at week 4 were 194.0 ± 15.44, 188.0 ± 19.24, 191.4 ± 12.89, 199.5 ± 16. 78 (grams), male strength 257.0 ± 18.31, 254.4 ± 37.52, 265.0 ± 19.75, 260.6 ± 24.12 (grams), food utilization rate (weight gain Z intake) Females are 21.99 ± 4.28, 20.98 people 3.28, 21.43 ± 3.26, 21.00 ± 2.23 (%), male strength 28.61 ± 2.41, 28.90 ± 7.12, 30, respectively 58 ± 2. 52, 31.05 ± 5.11 (%), hemoglobin level is 14.29 ± 0.76, 14.93 ± 0.91, 14.76 ± 0.68, 14.06 ± 0.67 (Gram Z deciliter;), male strength 14.67 ± 1.43, 14. 13 ± 1.42, 14. 17 ± 0.85, 14. 66 ± 0.66 (gram Z deciliter), white blood cell count is female strength S 15.87 ± 3. 35, 16.31 ± 4.27, 14.83 ± 3.76, 17.17 ± 4.00 (10 9th power Z liter), male power ^s 18.38 ± 3.69, 18.39 ± 3.24, 18.94 ± 4.01, 16.54 ± 1.67 (10 to the 9th power Honoré), Rinno ball ί Also, female force S respectively 83.4, 83.7, 81.4, 84.2 _^(0/0), Yuryoku S their respective 87.3, 88.6, 86.5, 86.3 ( Neutrophils, female strength 15.7, 15.8, 18.0, 15.2 (%), male strength 12.5, 11.0, 13.3, 13.4 (%) and other white blood cells, respectively. female force S respectively 0.9, 0.5, 0.6, 0.6 _^(0/0), Yuryoku S respectively 0. 2, 0.4, 0. 2, 0. 3 (%), GPT is female force respectively 54 . 1 ± 5.69, 64. 5 ± 11. 33, 51. 1 ± 6. 23, 4 5. 2 ± 4.87 ( international units Ζ liters), Yuryoku ^s respectively 58.4 ± 5. 17, 58. 7 ± 0. 33, 5 2.8 ± 5.07, 60.4 ± 4.43 (international unit Z liters), serum total protein is 82.5 ± 3.50, 90.9 ± 5.53, 80.9 ± 2. 33, 83.5 ± 4.40 (gram Z liter), male strength S 74.1 ± 3.84, 72.3 ± 3.13, 73.0 ± 4.14, 73.1 ± 3.90 (gram Z Liters), albumin is a female force S 39.1 ± 0.88, 39.2 ± 1.03, 39.0 people 0.67, 39. 1 ± 1.29 (gram Z liter), male strength ^s 36.6 ± 1.58, 35.8 ± 0.92, 36.1 ± 0.99, 36.0 ± 1.05 (gram / Lit Nore), triglyceride ί Female force 0.87 ± 0.22, 0.95 ± 0.15, 0.65 ± 0.08, 0.89 ± 0.25 (Midomo Mono / Ritsunare), male S 1.16 ± 0.49, 1.41 ± 0.50, 1.00 ± 0.21, 1.62 ± 0.54 (millimeter monolith / litnore), cholesterol, female force S, respectively 2.52 ± 0.34, 2.43 ± 0.34, 2.86 ± 0.35, 1.97 ± 0.29 (mmol liter), male power 2.24 ± 0.32, 1. 98 ± 0.18, 1.96 ± 0.16, 2.08 ± 0.32 (Midomole / Ritsunare), Gnore = f, female force ^s 4.76 ± 0.5, 4. 89 ± 0.87, 4.53 ± 0.39, 5.19 ± 1.06 (mmole Z liter), male power ^s 4.48 ± 0.58, 4.29 ± 0.63, 4 50 ± 0.57, 4.62 people 0.42 (mmol Z liter), urea is female force S 7.02 ± 1.12, 6.90 ± 0.81, 6.99 ± 0.65 7.96 ± 1.13 (mmol Z liter), male power ^s 5.80 ± 0.51, 5.75 ± 0.51, 4.87 ± 0.43, 5.25 ± 0.61 ( mmol Z l), liver body ratio, female force ^s their respective 4. 20 ± 0. 37, 4. 34 ± 0. 15, 4. 21 ± 0. 34, 3. 82 ± 0. 54 (% ), Male male 3.99 ± 0.34, 3.97 ± 0.37, 3.98 ± 0.16, 4.00 ± 0.15 (%) 0.89 ± 0.08, 0.80 ± 0.03, 0.81 ± 0.05, 0.83 ± 0.06 (%), male S respectively 0.83 ± 0.09, 0. 80 ± 0.04, 0.82 ± 0.04, 0.85 ± 0.05 (%). That is, compared to the control group that did not receive any drug, all drug-administered groups had body weight, food utilization, hemoglobin, white blood cell count, lymphocyte and neutrophil and other leukocyte ratios in white blood cells, GPT, and serum total protein. Serum albumin, triglycerides, cholesterol, dalcose, urea, liver ratio, and kidney ratio were not statistically significant.

 Example 7

 [0025] 25 healthy males and females weighing 25-30 grams each were divided into 5 groups, 5 males and 5 females, and the ACC hydrochloride monohydrate obtained in Example 4 was 0.0, 1. 25, 2.50, 5.00 (gram Z kilogram) and ethyl methanesulfonate 0.08 (gram Z kilogram) in 24 hour intervals. Group 0, 1.25, 2.50, 5.00, and MSE0.08) 6 hours later, the bone marrow of the sternum was removed and stained to examine the micronucleus rate. 0, 0, 1.25, 2.50, 5.00, MSEO.08 / J hemorrhoids rate is female force S, respectively 2.2, 1.4, 1.4, 1. 8, 34.2 (percentage) and male strength were 1.6, 1.8, 1.8, 1.6, 33.2 (percentage), respectively. There was no statistically significant difference in bone marrow micronucleus rate in any ACC hydrochloride monohydrate group compared to the control group that did not receive the drug. There was a statistically significant difference in the positive control group receiving ethyl methanesulfonate in the range of P 0.01 for both males and females.

 Example 8

[0026] 50 healthy male mice weighing 20-25 grams were divided into 10 groups and 5 groups, and the ACC hydrochloride monohydrate obtained in Example 4 was divided into 0, 1.25, 2.50. , 5.00 (gram Z kilogram weight), ma Itomycin C by 0.002 (gram Z kilogram body weight) for 5 days by gavage (each group in Kawagoe with 0.0 group, 1.25 group, 2.50 group, 5.00 group, MMC0.002 group) After 35 days, spermatozoa were also collected from the testes on both sides, and the rate of sperm malformation was examined. The sperm malformation rates of the 0.0, 1.25, 2.50, 5.00, and MMC0.002 groups were 1.58, 1.40, 1.36, 1.48, 6.10, respectively. (%)Met. There was no statistically significant difference in the rate of sperm malformation in any ACC hydrochloride monohydrate group compared to the control group that did not receive the drug. The positive control group receiving mitomycin C had a statistically significant difference in the range of P 0.01.

 Example 9

 [0027] In order to perform the Ames test, ACC hydrochloride monohydrate obtained in Example 4 was added by 5000, 1000, 200, 40, 8, and 0 microgram per petri dish, and Salmonella 4 strain TA 97a was added to the medium. TA98, TA100, and TA102 were separately inoculated and cultured at 37 degrees Celsius for 48 hours, and the number of colonies of mutant bacteria was counted. As a positive control group, a medium supplemented with 5 micrograms of the mutagen mitomycin C and 1.5 micrograms of NaN3 was used. Compared to the control group to which no drug was administered, the ACC hydrochloride monohydrate group had no statistically significant difference in the number of mutant colonies. The positive control group to which mitomycin C or NaN3 was administered had a statistically significant difference in the range of P 0.01.

 Example 10

[0028] Sixty healthy unmated female SD rats weighing 200-250 grams were divided into groups of 12 and 5 groups and fertilized. From day 7 to day 16 after fertilization, each day, the ACC hydrochloride monohydrate obtained in Example 4 was added in an amount of 0.0, 0.125, 0.25, 0.50 (gram Z kilogram body weight). Administration (each group in order: 0 group, 0.125 group, 0.25 group, 0.50 group), and the fertility rate was examined 20 days after fertilization. The fertility rates of the 0.0, 0.125, 0.25, and 0.50 groups were 75.00, 75.00, 91.67, and 83.33 (%), respectively. Body weight was measured on the fertilization day and 20 days after fertilization. The weight gain was 100.4 ± 28.59, 90.2 ± 20.74, 84.9 ± 24.57, 95.3 ± 14.31 (gram). The mortality rates were 0.00, 0.93, 0.98, and 0.00 (%), respectively. The sternum deletion rates reflecting fetal development were 49.02, 45.28, 47.92, and 39.62 (%), respectively. There were no statistically significant differences in the conception rate, weight gain after fertilization, mortality rate, or sternum loss rate in any ACC hydrochloride monohydrate group compared to the control group that did not receive the drug. Fetal weight, Compared with the control group, the height and tail length were also not significantly different.

 Example 11

 [0029] After obtaining 4-week-old male SHR-SPZlzm rats from the SHR and other disease model animal research group in Kyoto, the room temperature was 22 ± 1 degrees Celsius, the humidity was 60 ± 10%, and the lighting time was from 6 o'clock to 18 o'clock. Pre-bred for 1 week in a breeding room set at 12 hours until the time. Measure blood pressure and body weight of 40 rats with normal weight gain and no abnormalities in the general condition, so that 10 of these 40 rats were divided into 4 groups so that there was no difference in blood pressure or body weight between groups. divided. The following is a group in which 2.5% ACC hydrochloride monohydrate solution is injected by intraperitoneal injection of rats at a ratio of 50 mg Z kilogram body weight once daily, 5 times a week. The group that measures cerebral blood flow etc. is the B-ACC group, the group that administers the same amount of distilled water by intraperitoneal injection into rats is the control group, and the group that measures cerebral blood flow etc. of rats under the same administration conditions B-control group. During the experiment, they were bred with free intake of solid SP material and tap water. Regarding the handling of animal experiments, the “Basic Guidelines for Animal Experiments in the Physiological Field” established by the Physiological Society of Japan were followed.

 Example 12

 [0030] In Example 11, once a week, the rats were warmed for 10 minutes in a 38 ° C warmer before and after drug administration at the above temperature / humidity room at 11am and 5pm. After adaptation, the systolic phase was performed using the Softron non-invasive automatic blood pressure measuring device (BP-98A) (manufactured by Softron Co., Ltd.) by the tail-cuff method without anesthesia. Blood pressure, diastolic blood pressure and heart rate were measured three times, and the mean and standard deviation were calculated and expressed as mean standard error. The presence or absence of significant difference was examined by t-test.

The systolic blood pressure immediately after the start of administration of ACC hydrochloride monohydrate obtained in Example 4, 1 week, 2 weeks, 3 weeks, and 4 weeks after administration was 150.69 ± 11.2 for the control group, 175.38 ± 10.5, 199.58 ± 18.2, 207.50 ± 17.5, 213.61 ± 11.3 (millimeter Hg), ACC group power 144.77 ± 11.1, 153.33 ± 11.7 (* 3), 175.58 ± 15.2 (* 3), 194.41 ± 14.4 (*), 198.52 ± 16.7 (* 2) (mm Hg), and the diastolic blood pressure was 107.19 ± 11.7, 124.27 ± 10.2, 140.77 ± 17.0, 147.72 ± 8.69, 149.61 ± 13.68 in the control group, respectively. Mm Hg) and A CC group forces 104.22 ± 13.1, 111.33 ± 11.7 (*), 124.05 ± 9.6 (* 2), respectively , 146.22 ± 10.3, 150.11 ± 10.5 (mm), HR, HR, control group strength 359.83 ± 32.5, 384.05 ± 40.8, 349.80 ± 25.8, 359.66 ± 19.0, 349.58 ± 22.6 ACC group powers were 371.30 ± 27.0, 356.22 ± 29.3, 348.88 ± 29.8, 359.36 ± 22.6, and 354.61 ± 28.3 (apportioned number of times). The values in the ACC group that were significantly different in the range of Ρ <0.05, Ρ <0.01, and Ρ <0.001 compared to the control group are indicated by (*), (* 2), and (* 3), respectively. .

 Systolic blood pressure increased in both the ACC group and the control group until the 4th week of administration. In the ACC group, the blood pressure was significantly lower at 1, 2, 3, and 4 weeks after the start of administration compared to the control group. In addition, the blood pressure increase was suppressed (Ρ <0.001, 0.001, 0.05, 0.01). Regarding diastolic blood pressure, the ACC group showed significantly lower values than the control group at 1 and 2 weeks after the start of administration (Ρ <0.05 and Ρ <0.005, respectively), but there was a significant difference at 3 and 4 weeks. However, the power was not recognized. Regarding the heart rate, there was no significant difference between the ACC group and the control group at each week.

Example 13

In Example 12, after blood pressure and heart rate measurement 4 weeks after the start of ACC hydrochloride monohydrate administration, the rats were placed in a metabolic cage and collected for 24 hours. Concerning 24-hour excretion in control group and ACC group, Ν02 / Ν03 by Griess method, creatinine by alkaline picrin method, Na, K and C1 by ion electrode method, Mg by xylidyl-blue method, Ca by OCPC method Measured by N02 / N03 in control group and ACC group are 8.68 ± 3.26 and 4.92 ± 2.07 (*) (micro monore) for j jet, creatinine is 38.75 ± 7.92 and 35.87 ± 8.22 (milligram / decilit nore) for river page, Na is j jet 0.267 ± 0.086 and 0.396 ± 0.06 (water 2) (millimeter equivalent), K is 0.590 ± 0.17 and 0.669 ± 0.13 (millimeter equivalent) in Kawasega, C1 is 0.222 ± 0.07 and 0.31 in j jet 9 ± 0.08 (*) (millimeter equivalent), Mg was 1.61 ± 0.80 and 1.74 ± 0.63 (millimeter equivalent), and Ca was 0.20 ± 0.08 and 0.29 ± 0.11 (millimeter equivalent), respectively. . In the ACC group, N02ZN03 in urine was significantly lower than P <0.05 (*), compared with the control group, and urinary Na and C1 were P <0.005 (* 2) and P <0.05, respectively. The value was significantly high within the range of (*). There was no significant difference between creatine in AC C group and K, Mg, Ca compared with control group. NO02 / N03, which is a metabolite of NO, reflects the amount of NO excreted. NO, especially NO from eNOS is strong It is a powerful vasodilator. On the other hand, NO derived from N0, especially nN0S and iNOS, acts as a neuron-damaging substance. Three types of N0S inhibitors reduce the extent of brain tissue damage in stroke. The low N02ZN03 level in the ACC group seems to have reduced stroke, reduced brain damage, and extended life span in SHR-SP rats by reducing NO. However, the blood pressure lowering effect of ACC hydrochloride monohydrate appears to be related to other blood pressure regulators that have a weak relationship with NO. The amount of Na and C1 excretion was significantly higher in the ACC group than in the control group, because both groups were bred on the same diet, so it was possible to exert a blood pressure lowering effect by promoting NaCl excretion and preventing Na water retention. It is thought that there is sex.

Example 14

In Example 13, the urine was collected for 24 hours, then fasted for 24 hours, ether anesthetized, and blood was collected from the abdominal artery. Blood albumin by BCG method, albumin / globin ratio by calculation, GOT and GPT by UV method, gamma-daltamyl transpeptidase by colorimetric method, urea nitrogen by UV method, and Creathun by alkaline picrin method Na and K by ion electrode method, Ca by OCPC method, C1 by ion electrode method, Mg by xylidyl-blue method, angiotensin 1 transferase by Kasahara method, and angiotensin 2 by RIA 'Measured by DCC method. The blood albumin in the control group and ACC group were 2.31 ± 0.18 and 2.33 ± 0.15 (gram / decirit nore), respectively, and the anolebumin / globin ratio was 0.61 ± 0.02 and 0. 6 1 ± 0. 03 (calculated value), GOT is 476. 9 ± 178.1 and 532. 2 ± 175.7 (unit / liter), GPT is 115. 6 ± 43.9 And 126.5 ± 68.2 (unit / liter), gamma-gnoletoleminoretranspeptidase is 1.86 ± 0.90 and 3.17 ± 0.775 (* 2) (unit / liter) , Urea nitrogen in order 20.43 ± 2.82 and 19.50 ± 4.32 (milligram Z deciliter), creatinine in the j jet 0.37 ± 0.08 and 0.35 ± 0.06 (milligram / decirit nore ), Na on the river pages 148.0 ± 7.02 and 153.5 ± 10.8 (mm equivalent / Lit Nore), K on the j jet 6.36 ± 1.37 and 6.75 ± 0.67 (Milli equivalent / Little Nore), Ca is 10.95 ± 1.00 and 10.17 ± 2.69 (Milligram / Decirit Nore), and C1 is 104.0 ± 5.90 107. 3 ± 8.82 (milli-equivalent / Little Nore), Mg is 3.34 ± 0.45 and 2.85 ± 0.51 (Milligram / Decilit Nore) In order 28.47 ± 10. 3 and 25.53 ± 2.5 (unit Z liters), angiotensin 2 is in the river page 207.2 ± 20.4 and 150.2 ± 81.32 (picogram / millilitre) value Got. Blood albumin reflecting the nutritional status, albumin / golobulin ratio, and GOT, GPT, urea nitrogen, and Creathun, which represent liver and kidney function, all had no significant difference between the control group and the ACC group. The effects of salt monohydrate on the nutritional status and liver and kidney function are considered to be minimal. Gamma GTP level was significantly higher in the ACC group than in the control group within the range of P <0.001 (* 2), but within the normal range, it is unlikely to be a side effect of ACC hydrochloride monohydrate. . In addition, with regard to blood Na, Cl, K, Ca, and Mg levels, there was no significant difference between the ACC group and the control group. Indicates that there is no effect. Compared to the control group, the angiotensin 1 transferase and angiotensin 2 values in the ACC group tended to decrease. This may contribute to the blood pressure drop of ACC hydrochloride monohydrate.

 Example 15

 [0033] In Example 12, the incidence of stroke on the 100th day after administration of ACC hydrochloride monohydrate, the severity of the sequelae of stroke, and the survival status were observed and analyzed. Rats that died were dissected on the same day and pathologically examined for the presence of stroke lesions. The experimental data were expressed as mean standard error. The presence or absence of significant difference was examined by t-test.

 Up to 100 days after administration, the number of individuals who showed stroke symptoms from observation of behavioral neurological symptoms (hypersensitivity, hyperalgesia, limb paralysis, etc.) was 2 in the ACC group and 10 in the control group, respectively. AC C hydrochloride monohydrate administration was shown to suppress the onset of stroke. As for the number of surviving animals up to 100 days after administration, one animal in the ACC group died on the 79th day. In contrast, there were 3 deaths in the control group o, 2 in 75, 1 in 80, 3 in 82, 1 in 85, and 10 deaths. . A life-prolonging effect by administration of ACC hydrochloride monohydrate was confirmed (P 0.01). The anatomical macroscopic findings of the individuals who died showed infarcted lesions and diffuse cerebral edema and hemorrhagic lesions in the cerebral cortex and basal ganglia in all cases in the control group. Was light.

 Example 16

[0034] In Example 15, the severity was determined based on the behavioral neurological symptoms of the rat, and the score was evaluated. The following evaluation criteria were used. Sensation (painfulness, etc.) 1 point of hypersensitivity, one-limbed paralysis Power can walk independently, 2 points can be eaten voluntarily on the cage lid, there is one-limbed paralysis There were 3 points for the difficulty of voluntary walking, eating only the food in the cage, and 4 points for death. The ACC group had 4 points compared to the control group, and the ACC group significantly reduced the sequelae of stroke compared to the control group.

 Example 17

 [0035] In Example 11, on the 30th day after drug administration, continuous measurement and two-dimensional measurement of cerebral blood flow was performed using the laser-Doppler method on two rats in the B-ACC group and the B-control group. Measurements were made. As the measurement method, the rat scalp was incised under anesthesia, the skull was exposed, and the cerebral blood flow of the rat was measured using the laser power imaging device PiM2 (Risu Power Company, Sweden) manufactured by Rishiki. . The presence or absence of significant difference was examined by t-test. Continuous and two-dimensional measurements were performed using the laser Doppler method, and values of ACC group 1.464 ± 0.22 and control group 1.144 people 0.30 (P <0. 05) were obtained. This result shows that the cerebral blood flow situation of rats in the ACC group was improved from that in the control group.

 Example 18

 [0036] Risk factors for stroke development such as hypertension, diabetes, hyperlipidemia, heart disease, transient performance ischemic attack, possibility to be obtained by this trial for patients who want to participate in trial For patients who wish to participate after handing out a written statement that states that the benefits and risks of the trial can be withdrawn at any time and that the withdrawal will not be disadvantaged. In accordance with the clinical trial protocol approved in advance by the clinical trial ethics committee, there are two groups: the ACC group that receives ACC hydrochloride monohydrate on top of the basic treatment, and the control group that receives only basic treatment. Each patient's symptoms are scored according to the Japan 'Stroke' scale questionnaire, the score is calculated, and the onset of stroke is diagnosed with the calculated value according to the rules written in the questionnaire, and the incidence of stroke in the control group and ACC group From this difference, the stroke prevention effect of ACC hydrochloride monohydrate can be measured.

 The contents of the Japan 'Stroke' scale questionnaire are as follows.

 1. Consciousness: a) or b)

a) Glasgow · Koma · Scale: [Open eyes] 4 points to open eyes spontaneously, 3 points to open eyes by calling, 2 points to open eyes by pain stimulation, 1 point to never open eyes, [Language] 5 good, 4 confused conversations, 3 inappropriate words, 2 unintelligible responses, reaction 1 point for none, 6 points for following [exercise] commands, 5 points for appropriate response to pain, 4 points for flexion escape, 3 points for abnormal flexion response, 2 points for extension response, 1 point for no response, The total of [Open Eye], [Language], and [Exercise] is classified as 15 for A, 14-7 for B, and 6-3 for C.

 b) Japan 'Koma' scale: [Awakened without stimulating] 9 points for normality, clearness for general consciousness, 8 points for unclearness, 7 6 points for your name, your name, and your date of birth, 5 points to open your eyes easily with a normal call, 5 points to open your eyes by shaking your loud voice or body 4 points, pain ・ Simultaneously repeat the call while applying 3 points to barely open eyes, [not awakening even if stimulated, state] 2 points to move away from painful stimuli, painful stimulus In this case, 1 point is used to drive the limbs or frown, and 0 point is not to respond to pain stimulation. 9 points are classified as A, 8-3 points are classified as B, and 2-points are classified as C.

 The classification values (A = 7.74, B = 15.47, = 23.21) are used for the total.

 2. Language: Make a fist by verbal command, show the clock, say a clock, repeat the cherry, say the address' family name well: 4Z4 A, 3Z4 or 2Z4 B, 1/4 or 0 Classify Z4 as C, and use the classification values (A = l. 74, B = 2.95, C = 4.42) for the total.

 3. Ignore: A normal bisector test is A, a half-line ignorance is B in a bisector test, and I am aware of paralysis! /, Classifies actions that ignore the space on one side as C, and accumulates the classification values (A = 0.42, B = 0.85, C = l. 27).

 4. Visual field loss or semi-blindness: A homozygous visual field defect or no half-blind is classified as A, homogeneous visual field defect or semi-blind is classified as B, and classification values (A = 0.45, B = 0. 91) is used for the total.

 5.Eye movement disorder: Classify A as none, B as side view cannot be freely controlled, and C as unable to laterally view the other side without shifting to the eyeball, and the classification value (A = 0.84, B = l. 68, C = 2.53) is used for the total.

 6.Pupil abnormalities: Class A with no pupil abnormalities, B with pupil abnormalities on one side, C with abnormal pupils on both sides, and classification values (A = l. 03, B = 2.06, C = 3. 09) is used for the total.

 7.Facial paralysis: Classify none as A, shallow nose lips as B, resting mouth angle as the rest, and the classification values (A = 0.31, B = 0.62, C = 0.93) is accumulated.

8. Plantar reflexes: Classify normal as A, B as not normal, C as pathological reflex, and classify (A = 0.08, B = 0.15, C = 0.23) are accumulated.

 9. Sensory system: Normal is classified as A, mild sensory impairment is classified as B, distinct sensory impairment is classified as C, and classification values (A = —0.15, B = —0.29, C = 0. 44) is used for the total

10. Motor system:

 10- 1. Hand: Classify A as normal, make a ring with thumb and little finger, or hold a cup placed beside B, finger move but can't catch things or move at all, and classify as C Accumulate the values (A = 0.33, B = 0.66, C = 0.99).

 10-2.Arm: Normal A, arm can be raised with elbows extended, or arm can be raised by bending elbows B, arms move to some extent but cannot be lifted or do not move at all The classification values (A = 0.66, B = 1.31, C = 1.97) are used for the total.

 10-3.Lower limb hand: Class A is normal, B can be lifted with the knees stretched, or can stand on its own, B, Climbs can move but cannot stand, or no power at all The classification values (A = l. 15, B = 2.31, C = 3.46) are used for the total.

 If the value obtained by subtracting the constant 14.71 from the cumulative value of each category above is positive, it is determined that the stroke has occurred. From the difference in the incidence of stroke between the control group and the ACC group, the stroke prevention or treatment effect of ACC hydrochloride monohydrate can be measured.

Example 19

 Patients with sequelae of stroke are diagnosed according to the stroke motor dysfunction severity scale and grouped by severity. One group is divided into two groups: the ACC group that receives ACC hydrochloride monohydrate and the non-administered control group. In both groups, perform the same rehabilitation according to the severity of the serious injury, and regularly perform a scoring scale for the severity of motor impairment. The effect of ACC hydrochloride monohydrate on preventing or treating stroke sequelae is measured based on the difference in the severity of stroke motor dysfunction between the control group and the ACC group.

 The contents of the Stroke Motor Dysfunction Severity Scale are as follows.

 1. Facial paralysis: Classify A as “A” and “B” as “B”, and use the classification values (A = —1.27, B = l. 27) for the accumulation.

2. Dysphagia: None A, sometimes B, Tube 'feeding required Is classified as C, and the classification values (A = —4.93, B = −0.89, C = 5.82) are used for the total.

3.Arm: Class A can lift the arm with the elbow extended, B can lift the arm if the elbow is bent, C can not move against gravity, and the classification value (A = — 0.97, B = —0.09, C = 1.06) are used for the total.

 4.Hand: Classify A as normal, B to hold a cup placed beside, B as unpossible, C as classification value (A = —1.26, B = —0.16, C = 1. 42) is used for the total.

 5. Proximal muscles of the lower limbs: Examined in the supine position, classifying normal as A, knee-capable as B, and non-capable as C, classification values (A = —1.04, B = 0.14, C = 0. 89) is used for the total.

 6.Ankle joint: Examined in the sitting position, and if the sitting position cannot be estimated, it is estimated from the muscle strength in the supine position, classifying the toe up as A and classifying out the toe as B, the classification value (A =-0 52, B = 0.5 2) is used for the total.

 7. Combined exercise: Supine position on the bed A series of movements up to the bedside position, classifying A as standing beside the bed, B as sitting in the bed, C as not sitting, and the value of the classification (A = — 1. 2 4, B = —0. 39, C = 1. 63).

 8.Walking: Classify as A (can walk without assistive devices, B as walkable with assistive devices or caregivers, C as unable to walk on their own, and classification values (A = —3.63, B = —0) 45, C = 4.08) is used for the total.

 A constant of 14.60 is added to the cumulative value of each category above, and if the value is positive, it is determined that there is sequelae of stroke. From the difference in the severity of stroke motor dysfunction between the control group and the ACC group, the effect of preventing or treating stroke sequelae of ACC hydrochloride monohydrate can be measured.

Industrial applicability

 The drug of the present invention can be used as a preventive or therapeutic agent for stroke or stroke sequelae.

Claims

 The scope of the claims  1-Aminocyclopropanecarboxylic acid and prodrugs thereof, and pharmaceutically acceptable salts and hydrates thereof A drug for the prevention or treatment of stroke or stroke sequelae comprising a selected compound as a main component .  The medicament for preventing or treating stroke according to claim 1.