JP2018080118A - Injury therapeutic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel injury therapeutic agent having the action to inhibit the leakage of a blood component from an injury.SOLUTION: An injury therapeutic agent contains cyclic phosphatidic acid, carba-cyclic phosphatidic acid or thia-cyclic phosphatidic acid, or a salt thereof.SELECTED DRAWING: None

Description

  The present invention relates to a therapeutic agent for injury comprising cyclic phosphatidic acid, carbacyclic phosphatidic acid or thiacyclic phosphatidic acid as an active ingredient.

  In 1992, a monophasic myxomycoba of the true slime mold Physarum polycephalum was found to be a fat-soluble substance that suppresses the activity of DNA polymerase α, a DNA replication enzyme of eukaryotic cells, and suppresses the growth of cultured animal cells. Separated and purified (Non-patent Document 1)). This substance is an LPA-like substance derived from Physarum, which is found to be a substance in which hexadecanoic acid containing cyclopropane is bonded to the sn-1 position of the glycerol skeleton and phosphoric acid is bonded to the 2nd and 3rd positions by a cyclic ester bond. And was named PHYLPA. Since PHYLPA has a characteristic fatty acid at the sn-1 position, chemical synthesis of a derivative substituted with a general fatty acid and investigation of its activity showed that it suppresses cell growth and suppresses PHYLPA growth. It was revealed that the action was due to the cyclic phosphate groups at the 2nd and 3rd positions. At present, LPA analogs having such a cyclic phosphate group are collectively called cyclic phosphatidic acid (cPA).

  Regarding cyclic phosphatidic acid and its derivatives, neurotrophic factor action and application to neurodegenerative diseases (Patent Documents 1 and 2), suppression of proliferation and invasion / metastasis of cancer cells (Patent Document 3), analgesic action (Patent Document 4) ) As well as application to atopic dermatitis (Patent Document 5) and application to demyelinating disease (Patent Document 6 and Non-Patent Document 2), but there is no knowledge about the effect on damage treatment.

JP 2002-308778 A JP 2002-308879 A International Publication WO2002 / 94286 International Publication WO2008 / 81580 JP 2012-56853 A International Publication No. WO2014 / 115585

Murakami-Murofushi, K., et al., J. Biol. Chem. 267, 21512-21517 (1992) Yamamoto, S., et al., European Journal of Pharmacology 741 (2014) 17-24

  If a drug having an action of suppressing the leakage of blood components from damage such as traumatic injury can be found, a novel therapeutic agent for damage can be developed. In particular, the repair mechanism of traumatic brain injury has not been fully clarified, and no therapeutic agent for traumatic brain injury has been found. An object of the present invention is to provide a novel therapeutic agent for damage that has an action of suppressing leakage of blood components from damage.

  The present inventors have found that cyclic phosphatidic acid, carbacyclic phosphatidic acid or thiacyclic phosphatidic acid and derivatives thereof have an action of suppressing leakage of blood components from damage, and have completed the present invention.

That is, according to the present invention, there is provided an injury treatment agent containing cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or a salt thereof.
Preferably, the cyclic phosphatidic acid, the carbacyclic phosphatidic acid or the thiacyclic phosphatidic acid is a compound represented by the formula (1).
(In the formula, R is a linear or branched alkyl group having 1 to 30 carbon atoms, a linear or branched alkenyl group having 2 to 30 carbon atoms, or a linear or branched group having 2 to 30 carbon atoms. An alkynyl group, which may contain a cycloalkane ring or an aromatic ring, wherein X and Y each independently represent —O—, —S— or —CH ₂ —, at the same time -CH ₂ - not to become that .M represents a hydrogen atom or a counter cation).

Preferably, in Formula (1), one of X and Y is —CH ₂ —, and the other of X and Y is —O—.
Preferably, the injury is a traumatic injury.
Preferably, the injury is traumatic brain injury.
Preferably, the therapeutic agent for injury of the present invention is used as a hemostatic agent having a hemostatic action.

  According to the present invention, there is provided a method of treating an injury comprising administering a cyclic phosphatidic acid, a carbacyclic phosphatidic acid or a thiacyclic phosphatidic acid or a salt thereof to a patient having an injury.

The present invention further provides the use of cyclic phosphatidic acid, carbacyclic phosphatidic acid or thiacyclic phosphatidic acid or a salt thereof for the manufacture of a therapeutic agent for injury.
The present invention further provides cyclic phosphatidic acid, carbacyclic phosphatidic acid or thiacyclic phosphatidic acid or salts thereof for use in the treatment of damage.

  ADVANTAGE OF THE INVENTION According to this invention, the damage therapeutic agent characterized by containing cyclic phosphatidic acid, carbacyclic phosphatidic acid, or thiacyclic phosphatidic acid as an active ingredient can be provided.

FIG. 1 shows the result of observing blood leakage from an injured part to brain tissue as an index of damage repair using a color image of IgG. FIG. 2 shows the results of quantifying the color development of IgG using ImageJ.

Hereinafter, the present invention will be described more specifically.
The present invention relates to a therapeutic agent for injury containing cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or a salt thereof as an active ingredient. The injury therapeutic agent of the present invention can be used to treat injury, and contains cyclic phosphatidic acid carbacyclic phosphatidic acid or thiacyclic phosphatidic acid, or a salt thereof as an active ingredient. The cyclic phosphatidic acid, the carbacyclic phosphatidic acid, or the thiacyclic phosphatidic acid is not particularly limited as long as it exhibits the effects of the present invention. Preferably, a cyclic phosphatidic acid represented by the following formula (I) can be used. .

(In the formula, R is a linear or branched alkyl group having 1 to 30 carbon atoms, a linear or branched alkenyl group having 2 to 30 carbon atoms, or a linear or branched group having 2 to 30 carbon atoms. An alkynyl group, which may contain a cycloalkane ring or an aromatic ring, wherein X and Y each independently represent —O—, —S— or —CH ₂ —, at the same time -CH ₂ - not to become that .M represents a hydrogen atom or a counter cation).

  In the formula (I), specific examples of the linear or branched alkyl group having 1 to 30 carbon atoms represented by the substituent R include, for example, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, and a hexyl group. , Heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl and the like.

  Specific examples of the linear or branched alkenyl group having 2 to 30 carbon atoms represented by the substituent R include, for example, an allyl group, a butenyl group, an octenyl group, a decenyl group, a dodecadienyl group, a hexadecatrienyl group, and the like. More specifically, 8-decenyl group, 8-undecenyl group, 8-dodecenyl group, 8-tridecenyl group, 8-tetradecenyl group, 8-pentadecenyl group, 8-hexadecenyl group, 8-heptadecenyl group, 8- Octadecenyl group, 8-icosenyl group, 8-docosenyl group, heptadec-8,11-dienyl group, heptadec-8,11,14-trienyl group, nonadeca-4,7,10,13-tetraenyl group, nonadeca-4, 7,10,13,16-pentaenyl group, henicosa-3,6,9,12,15,18-hexaenyl group, etc. It is.

  Specific examples of the linear or branched alkynyl group having 2 to 30 carbon atoms represented by the substituent R include, for example, 8-decynyl group, 8-undecynyl group, 8-dodecynyl group, 8-tridecynyl group, and 8-tetradecynyl group. Group, 8-pentadecynyl group, 8-hexadecynyl group, 8-heptadecynyl group, 8-octadecynyl group, 8-icosinyl group, 8-docosinyl group, heptadec-8,11-diynyl group and the like.

  Specific examples of the cycloalkane ring that can be contained in the alkyl group, alkenyl group, or alkynyl group include a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, and a cyclooctane ring. The cycloalkane ring may contain one or more heteroatoms, and examples thereof include an oxirane ring, an oxetane ring, a tetrahydrofuran ring, and an N-methylprolysine ring.

  Specific examples of the aromatic ring that can be contained in the alkyl group, alkenyl group, or alkynyl group include a benzene ring, a naphthalene ring, a pyridine ring, a furan ring, and a thiophene ring.

  Therefore, specific examples when the substituent R is an alkyl group substituted with a cycloalkane ring include, for example, a cyclopropylmethyl group, a cyclohexylethyl group, an 8,9-methanopentadecyl group, and the like.

Specific examples when the substituent R is an alkyl group substituted by an aromatic ring include a benzyl group, a phenethyl group, a p-pentylphenyloctyl group, and the like.

  R is preferably a linear or branched alkyl group having 9 to 17 carbon atoms, a linear or branched alkenyl group having 9 to 17 carbon atoms, or a linear or branched alkynyl group having 9 to 17 carbon atoms. It is a group. R is more preferably a linear or branched alkyl group having 9, 11, 13, 15 or 17 carbon atoms, or a linear or branched alkenyl group having 9, 11, 13, 15 or 17 carbon atoms. is there. R is particularly preferably a linear or branched alkenyl group having 9, 11, 13, 15 or 17 carbon atoms.

X and Y in the compound represented by the general formula (1) each independently represent —O—, —S— or —CH ₂ —, but X and Y are not simultaneously —CH ₂ —. That is, there are the following three combinations of X and Y.
(1) X is —O— and Y is —O—.
(2) X is —CH ₂ — and Y is —O— (2-carba cPA (also abbreviated as 2ccPA)). Or X is -S- and Y is -O-.
(3) X is —O— and Y is —CH ₂ — (3-carba cPA (also abbreviated as 3ccPA)). Or X is -O- and Y is -S-.
Among the above, (2-carba cPA) in which X is —CH ₂ — and Y is —O— is particularly preferable.

  M in the cyclic phosphatidic acid derivative represented by the formula (I) is a hydrogen atom or a counter cation. Examples of the case where M is a counter cation include, for example, an alkali metal atom, an alkaline earth metal atom, and a substituted or unsubstituted ammonium group. Examples of the alkali metal atom include lithium, sodium, and potassium, and examples of the alkaline earth metal atom include magnesium and calcium. Examples of the substituted ammonium group include a butyl ammonium group, a triethyl ammonium group, and a tetramethyl ammonium group.

  The compounds of formula (I) may have isomers such as positional isomers, geometric isomers, tautomers, or optical isomers depending on the type of substituent, but all possible Isomers and mixtures containing two or more of these isomers in any ratio are also within the scope of the present invention.

  The compound of formula (I) may exist in the form of an adduct (hydrate or solvate) with water or various solvents, and these adducts are also within the scope of the present invention. is there. Furthermore, any crystalline form of the compound of formula (I) and salts thereof is within the scope of the invention.

  Preferably, the compound represented by the formula (1) is 1-oleoyl cyclic phosphatidic acid or 1-palmioleoyl cyclic phosphatidic acid. Specific examples of the compound represented by the formula (1) used in the present invention include oleoyl 2-carba cPA (ΔOle-2ccPA), palmitooleoyl 2-carba cPA (ΔPal-2ccPA) and the like.

  Among the compounds represented by the formula (1), compounds in which X and Y are —O— are, for example, Kobayashi, S., et al .: Tetrahedron Lett., 34, 4047-4050 (1993), JP-A-5-230088. It can be chemically synthesized according to the methods described in JP-A-7-149772, JP-A-7-258278, and JP-A-9-25235.

Further, among the compounds represented by the formula (1), a compound in which X and Y are —O— can be obtained by allowing phospholipase D to act on lyso-type phospholipid according to the method described in JP-A No. 2001-178489. It can also be synthesized. The lyso-type phospholipid used here is not particularly limited as long as it is a lyso-type phospholipid capable of acting on phospholipase D. Many types of lyso-type phospholipids are known, those having different fatty acid types, and molecular types having ether or vinyl ether bonds are known, and these are available as commercial products. As phospholipase D, those derived from higher plants such as cabbage and peanuts, and those derived from microorganisms such as Streptomyces chromofuscus and Actinomadula sp. Are available as commercially available reagents, but are extremely selected by the enzyme derived from Actinomadula sp. CPA is synthesized (Japanese Patent Laid-Open No. 11-367032). The reaction between lyso-type phospholipid and phospholipase D is not particularly limited as long as the enzyme can exhibit its activity. For example, it is heated from room temperature in an acetic acid buffer solution (about pH 5 to 6) containing calcium chloride. (Preferably at about 37 ° C.) for about 1 to 5 hours. The produced cPA derivative can be purified by extraction, column chromatography, thin layer chromatography (TLC) or the like according to a conventional method.

  Further, among the compounds represented by the formula (1), a compound in which X or Y is —S— is represented by Bioorganic & Medicinal Chemistry Letters 21 (2011) 4180-4182 or Bioorganic & Medicinal Chemistry 20 (2012) 3196-3201. It can be synthesized according to the description.

Further, among the compounds represented by the formula (1), a compound in which X is —CH ₂ — and Y is —O— is a method described in Japanese Patent Application Laid-Open No. 2004-010582 or International Publication WO 03/104246. Can be synthesized.

Further, among the compounds represented by the formula (1), a compound in which X is —O— and Y is —CH ₂ — can be obtained by a method described in the literature (Uchiyama A. et al., Biochimica et Biophysica Acta 1771 (2007 103-112; and “The Pharmaceutical Society of Japan 23rd Symposium on Progress in Reactions and Synthesis, November 17 and 18, 1997 (Kumamoto Civic Center) Synthesis and Physiological Actions of Cyclic Phosphatidic Acid and Carba Derivatives, Abstracts Page 101- 104 ") and can be synthesized by the method described in International Publication No. WO2002 / 094286. An example of a specific synthesis route is shown below.

In the above, first, the commercially available (R) -benzyl glycidyl ether (1) is activated with BF ₃ .Et ₂ O, and the thio compound obtained by reacting n-BuLi with methylphosphonic acid dimethyl ester is reacted. To obtain alcohol (2).
The obtained alcohol is reacted at 80 ° C. with an excess of pyridinium salt of p-toluenesulfonic acid in toluene to obtain a cyclized product (3). This cyclized product is subjected to hydrogenolysis using 20% Pd (OH) ₂ —C under hydrogen atmosphere to perform debenzylation (4). Using 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride as a condensing agent, it is reacted with a fatty acid to obtain a coupled product (5). Next, using bromotrimethylsilane as a nucleophile, only the methyl group is regioselectively removed to obtain cyclic phosphonic acid (6). This is transferred to a separatory funnel using ether, and a small amount of 0.02N aqueous sodium hydroxide solution is added dropwise to carry out a liquid separation operation to extract and purify the target compound as a sodium salt (7).

  Cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or a salt thereof used as an active ingredient in the present invention has an action of suppressing the leakage of blood components from damage such as traumatic injury, so that it is a therapeutic agent for damage (damage It can be used as a restorative agent). In particular, cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or salts thereof used as active ingredients in the present invention can be used as a hemostatic agent by having a hemostatic action at the damaged site (wound).

  Examples of the damage to be administered with the damage treatment agent (damage repair agent) of the present invention include central nerve damage (for example, brain damage such as traumatic brain damage or spinal cord damage), tissue damage, organ damage (liver damage, spleen). Injury, pancreatic damage, kidney damage, gastrointestinal damage, mesentery, omentum, omental damage, thorax damage, trachea / bronchial damage, lung damage, diaphragm damage, heart damage, large blood vessel damage, pelvic damage, etc.), skin damage (For example, but not limited to, burns, lacerations, wounds, punctures, or traumas). Among these, the injury therapeutic agent of the present invention is particularly useful as a therapeutic agent for traumatic brain injury.

  The therapeutic agent for injury of the present invention comprises one or more pharmaceutically acceptable pharmaceutical additives and an active ingredient cyclic phosphatidic acid, carbacyclic phosphatidic acid or thiacyclic phosphatidic acid (preferably represented by the formula (1) And a salt thereof are preferably provided in the form of a pharmaceutical composition.

  The agent for treating damage according to the present invention can be administered in various forms, and suitable administration forms include oral administration and parenteral administration (for example, intravenous, intramuscular, subcutaneous or intradermal injection, (Intrarectal administration, transmucosal administration, etc.) may be used, but parenteral administration is preferred, and local administration to the site of injury is particularly preferred.

  Examples of the pharmaceutical composition suitable for oral administration include tablets, granules, capsules, powders, solutions, suspensions, syrups, etc. Examples of the pharmaceutical composition suitable for parenteral administration include , Injections, infusions, suppositories, transdermal absorption agents, and the like, but the dosage form of the therapeutic agent for injury of the present invention is not limited thereto. Furthermore, it can also be set as a continuous formulation by a well-known technique. For example, a sustained-release preparation can be obtained by encapsulating cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or a salt thereof as an active ingredient in a hydrogel based on gelatin.

  There are no particular limitations on the type of formulation additive used in the production of the injury treatment agent of the present invention, and a person skilled in the art can appropriately select it. For example, excipients, disintegrants or disintegration aids, binders, lubricants, coating agents, bases, solubilizers or solubilizers, dispersants, suspending agents, emulsifiers, buffers, antioxidants, antiseptics Agents, isotonic agents, pH adjusters, solubilizers, stabilizers, and the like can be used, and the specific components used for these purposes are well known to those skilled in the art.

  Pharmaceutical additives that can be used in the preparation of formulations for oral administration include, for example, excipients such as glucose, lactose, D-mannitol, starch, or crystalline cellulose; carboxymethylcellulose, starch, carboxymethylcellulose calcium, etc. Disintegrating agents or disintegrating aids; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or gelatin; lubricants such as magnesium stearate or talc; coatings such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol, or titanium oxide Agents: Bases such as petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, or hard fat can be used.

  Examples of pharmaceutical additives that can be used for the preparation of pharmaceutical preparations for injection or infusion include aqueous solutions such as distilled water for injection, physiological saline, propylene glycol, surfactants, etc. Agents or solubilizers; isotonic agents such as glucose, sodium chloride, D-mannitol, glycerin, etc .; use of pharmaceutical additives such as pH regulators such as inorganic acids, organic acids, inorganic bases or organic bases it can.

The therapeutic agent for injury of the present invention can be administered to mammals such as humans.
The dose of the therapeutic agent for injury according to the present invention should be appropriately increased or decreased depending on conditions such as the age, sex, weight, symptoms, and administration route of the patient. Is in the range of about 1 μg / kg to 1,000 mg / kg, and preferably in the range of about 10 μg / kg to 100 mg / kg. The above-mentioned dose of the therapeutic agent for injury may be administered once a day, or may be administered in several times (for example, about 2 to 4 times).

  The present invention will be specifically described by the following examples, but the present invention is not limited to the examples.

Example 1:
(1) Test compound Carba cPA used in the experiment is 2ccPA (18: 1). The structure of 2ccPA (18: 1) is as follows. Here, —C ₁₇ H ₃₃ represents — (CH ₂ ) ₇ CH═CH (CH ₂ ) ₇ CH ₃ (cis isomer). 2ccPA (18: 1) was synthesized by the method described in JP-A-2004-010582.

(2) Mouse
Six-week-old female ICR mice (Charles River) were raised in an animal breeding room using a plastic cage. The breeding environment was a constant temperature of 22 ° C., and the light and dark were cycled 12 hours. The experiment was approved by the Ministry of Education, Culture, Sports, Science and Technology and the Ochanomizu University Animal Experiment Committee, and was conducted according to these guidelines.

(3) Brain injury The puncture injury was performed on 6-week-old female mice. Three anesthetics (0.75 mg / kg domitol (Nippon Zenyaku Kogyo), 4.0 mg / kg midazolam (sand), 5.0 mg / kg betorfal (Meiji Seika Pharma)) were intraperitoneally administered to mice, and then the occipital region The skull was exposed, and a hole was made in the skull on the right brain side of the occipital region using a syringe needle (Terumo needle 19G × 1 1/2 ”, Terumo). Next, the needle (Terumo needle 27G × 3/4) ", Terumo) was used to puncture the cerebral cortex horizontally from the skull hole. After the stab wound, an anti-anedan against anesthesia (0.75 mg / kg) (Nippon Zenyaku Kogyo) was administered intraperitoneally. The left brain was treated as a control experiment. 500 μg / ml 2ccPA was intraperitoneally administered at a dose of 1.8 mg / kg / day every 24 hours immediately after the injury. As a Control group, a phosphate buffer solution (PBS; 0.135 M NaCl, 8.1 mM Na ₂ HPO ₄ , 2.685 mM KCl, 1.47 mM KH ₂ PO ₄ ) was administered intraperitoneally.

(4) Fixation After puncture injury 1, 3, 5, and 7 days, mice were euthanized using CO ₂ , 50 ml of PBS, fixative (4% paraformaldehyde, 80 mM Na ₂ HPO ₄ , 20 mM NaH ₂ PO ₄ ) Injection into the left ventricle in the order of 50 ml, and perfusion fixation was performed. After removing the whole brain, it was placed in a fixative and shaken overnight at room temperature. The fixative was replaced with 15% sucrose and shaken overnight at 4 ° C., and then replaced with 30% sucrose and shaken overnight at 4 ° C.

(5) Preparation of embedding / frozen tissue section Embedding agent (OCT Compound) (Sakura Finetech) is placed in cryomold (TED PELLA, Inc.) containing whole brain, shaken at 4 ° C for 30 minutes, and dried ice And the whole brain were frozen in a foamed polystyrene box containing isopentane and stored at -80 ° C. A frozen tissue section was prepared using a cryostat section preparation apparatus cryostat (CM1850, Leica) and attached to a slide glass (Superfrost Plus, Fisher Scientific). The section was a coronal section having a thickness of 20 μm.

(6) Immunohistochemical staining using IgG As an index for repairing damage, leakage of blood from the damaged part to the brain tissue was observed. Enzyme antibody staining with anti-mouse IgG antibody was performed on coronal sections frozen sections of mouse brain 1, 3, 5, and 7 days after puncture injury, and blood leakage was visualized by coloring IgG. Furthermore, the amount of blood leakage in the 2ccPA administration group and the Control group was analyzed from the color intensity. The method of enzyme antibody staining is shown below. First, frozen coronal sections were dried at 37 ° C. for 1 hour. All subsequent operations were performed at room temperature. The solution was fixed with a fixative for 30 minutes, washed with PBS for 5 minutes × 2 times, and with Tris buffer (TBS; 25 mM Tris, 137 mM NaCl, 2.68 mM KCl, pH 7.4) for 5 minutes × 2 times. A TBS mixed solution containing 10% methanol and 3% H ₂ O ₂ was added and reacted for 10 minutes to inactivate endogenous peroxidase, and then washed with TBS for 5 minutes × 3 times. Thereafter, blocking was performed for 1 hour using a blocking solution (10% calf serum, 3% bovine serum albumin, 133 mM glycine, 0.4% Triton X-100: in TBS). Next, biotin-labeled anti-mouse IgG antibody (AP124B, Merck Millipore) was diluted with a blocking solution and allowed to react for 1 hour. Furthermore, after forming an avidin-biotin labeled enzyme complex by reacting with VECTASTAIN Elite ABC Standard Kit reagent (Vector Laboratories) for 1 hour, it was washed 5 times x 3 times with 0.1M Tris-HCl (pH 8.0) . The color was developed by reacting overnight with a mixed solution of 0.05% diaminobenzidine, 0.003% H ₂ O ₂ , 0.1M Tris-HCl (pH 8.0). The next day, after washing with distilled water, 70% ethanol, 95% ethanol, 100% ethanol, and xylene were replaced in this order, and finally sealed with Canadian balsam.

(7) Observation and quantification
Enzyme antibody-stained images after 1, 3, 5, and 7 days after puncture injury in the 2ccPA administration group and the Control group were observed and photographed in a bright field using an inverted microscope (FSX-100, Olympus). The results are shown in FIG.
The color development intensity of IgG was quantified using ImageJ as follows. First, the image photographed in each section was converted to 8 bits to obtain a grayscale image. Furthermore, the black and white were reversed. Thereafter, five regions (8 × 8 μm ² ) around the right brain injury region were randomly selected, and the mean gray value in each region was measured. In addition, five regions were selected from the cerebral cortex region of the left brain that was not damaged, and the mean gray value was measured. In order to remove the background of staining, the mean gray value of the left brain side was subtracted from the right brain side, and the value was used as the color intensity of IgG. The above operation was performed on sections (8 to 10 pieces per individual) including the damaged sites of the whole brain extracted from each individual. The average value of the color development intensity of IgG obtained from each section is taken as the value of that individual, and the results of averaging the values of three individuals are shown in FIG.

(8) Result By intraperitoneal administration of 2ccPA after puncture injury, the amount of blood leakage was significantly 28% on the 1st day, 34% on the 3rd day, 42% on the 5th day, 60% on the 7th day compared with the Control group It was decreasing. This indicates that 2ccPA suppresses blood leakage and promotes repair.

Claims (6)

A therapeutic agent for injury comprising cyclic phosphatidic acid, carbacyclic phosphatidic acid, thiacyclic phosphatidic acid or a salt thereof. The injury therapeutic agent according to claim 1, wherein the cyclic phosphatidic acid, the carbacyclic phosphatidic acid or the thiacyclic phosphatidic acid is a compound represented by the formula (1).
(In the formula, R is a linear or branched alkyl group having 1 to 30 carbon atoms, a linear or branched alkenyl group having 2 to 30 carbon atoms, or a linear or branched group having 2 to 30 carbon atoms. An alkynyl group, which may contain a cycloalkane ring or an aromatic ring, wherein X and Y each independently represent —O—, —S— or —CH ₂ —, at the same time -CH ₂ - not to become that .M represents a hydrogen atom or a counter cation). In the formula (1), one of X and Y is -CH ₂ -, the other of X and Y is -O-, and damaging therapeutic agent according to claim 2. The damage therapeutic agent according to any one of claims 1 to 3, wherein the damage is traumatic damage. The injury therapeutic agent according to any one of claims 1 to 4, wherein the injury is traumatic brain injury. The injury therapeutic agent according to any one of claims 1 to 5, which is used as a hemostatic agent having a hemostatic effect.