WO2022050355A1 - ドコサヘキサエン酸、ドコサヘキサエノイル基含有ホスファチジン酸、又はその誘導体によるセロトニントランスポーター関連精神疾患の予防・治療剤 - Google Patents

ドコサヘキサエン酸、ドコサヘキサエノイル基含有ホスファチジン酸、又はその誘導体によるセロトニントランスポーター関連精神疾患の予防・治療剤 Download PDF

Info

Publication number
WO2022050355A1
WO2022050355A1 PCT/JP2021/032309 JP2021032309W WO2022050355A1 WO 2022050355 A1 WO2022050355 A1 WO 2022050355A1 JP 2021032309 W JP2021032309 W JP 2021032309W WO 2022050355 A1 WO2022050355 A1 WO 2022050355A1
Authority
WO
WIPO (PCT)
Prior art keywords
praja
activity
group
acid
obsessive
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2021/032309
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
郁夫 坂根
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chiba University NUC
Original Assignee
Chiba University NUC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chiba University NUC filed Critical Chiba University NUC
Priority to JP2022546971A priority Critical patent/JPWO2022050355A1/ja
Publication of WO2022050355A1 publication Critical patent/WO2022050355A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • A23L33/12—Fatty acids or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/661—Phosphorus acids or esters thereof not having P—C bonds, e.g. fosfosal, dichlorvos, malathion or mevinphos
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing

Definitions

  • the present invention relates to drugs such as prophylactic and therapeutic agents for the control of serotonin / 5-hydroxytryptamine (5-HT) transporter (SERT), obsessive-compulsive disorder (OCD), major depressive disorder, autism, integration. It relates to drugs for the prevention and treatment of ataxia, anxiety, depression, obsessive-compulsive disorder, and impulsive psychiatric disorders, as well as foods having such effects.
  • drugs such as prophylactic and therapeutic agents for the control of serotonin / 5-hydroxytryptamine (5-HT) transporter (SERT), obsessive-compulsive disorder (OCD), major depressive disorder, autism, integration. It relates to drugs for the prevention and treatment of ataxia, anxiety, depression, obsessive-compulsive disorder, and impulsive psychiatric disorders, as well as foods having such effects.
  • 5-HT 5-hydroxytryptamine
  • OCD obsessive-compulsive disorder
  • Serotonin / 5-hydroxytryptamine (5-HT) is known to be involved in anxiety, depression, obsessive-compulsiveness, and impulsivity [Non-Patent Document 1].
  • Serotonin / 5-HT transporter (SERT) reabsorbs 5-HT from synaptic clefts into presynaptic neurons for recycling and metabolic degradation [Non-Patent Document 2; Non-Patent Document 3].
  • Selective serotonin reabsorption (SERT) inhibitors (SSRIs) are used to treat obsessive-compulsive disorder (OCD) [Non-Patent Document 2; Non-Patent Document 3] and major depressive disorder [Non-Patent Document 4].
  • Non-Patent Document 5 Several studies with SERT-deficient rodents have demonstrated that SERT is associated with neurodevelopmental disorders such as depression, anxiety, and autism [Non-Patent Document 5]. Thus, the hyperactivity of SERT, which reduces the amount of 5-HT in the synaptic cleft, causes the various psychiatric disorders described above. However, the control mechanism of the SERT function is still not well understood.
  • DG Diacylglycerol
  • ⁇ - ⁇ Diacylglycerol
  • PA phosphatidic acid
  • DG and PA are established lipid second messengers, which control a wide variety of physiological and pathological events.
  • the present inventors have generated and analyzed brain-specific DGK ⁇ knockout (KO) mice, and have revealed that KO mice exhibit SSRI (fluoxetine) -sensitive OCD-like behavior [Non-Patent Document 11].
  • DGK ⁇ As SERT [Non-Patent Document 12; Non-Patent Document 13], melanoma antigen gene-D1 (MAGE-D1) [Non-Patent Document 13], and Praja-1 E3 ubiquitin.
  • -Protein ligase (This Praja-1 E3 ubiquitin-Protein ligase ubiquitinizes SERT [Non-Patent Document 14], and Praja-1-ubiquitin-proteasome system in a DGK activity-dependent manner. It was found that it interacts with [Non-Patent Document 13]), which induces SERT degradation mediated by. However, it remains unclear how Praja-1 activity is controlled by DGK ⁇ .
  • Serotonin transporter controls the serotonergic system, as well as serotonin / SERTs such as obsessive-compulsive disorder (OCD), depression, autism, and schizophrenia. Involved in the pathophysiology / therapeutics of related diseases.
  • SERT inhibitors SERT inhibitors
  • SSRIs SERT inhibitors
  • SERT-related diseases for example, depression, obsessive-compulsive disorder (OCD), schizophrenia, Alzheimer-type dementia, etc.
  • DG diacylglcerol
  • DGK diacylglcerol
  • DGK diacylglcerol
  • Praja-1 Praja-1 E3 ubiquitin-protein ligase
  • SERT is controlled to make serotonin / SERT-related diseases such as obsessive-compulsive disorder (OCD), major depressive disorder, autism, schizophrenia, anxiety, depression, obsessive-compulsive disorder, and impulsiveness. It is required to find a drug useful for the prevention and treatment of psychiatric disorders, and a drug having a similar effect and activity that can be easily ingested as a food.
  • OCD obsessive-compulsive disorder
  • impulsiveness impulsiveness
  • the present inventors have pursued research focusing on the molecular mechanism by which DGK ⁇ regulates Praja-1 activity.
  • PA phosphatidic acid
  • Y carbon atom of fatty acid acyl portion of gly
  • the present inventors have also found that 18: 0/22: 6-PA selectively interacts with Praja-1 to enhance its E3 ubiquitin-protein ligase activity. These results strongly suggest that DGK ⁇ activates Praja-1 and degrades SERT through 18: 0/22: 6-PA production. Our findings provide new insights into SERT level control and the pathophysiology / therapeutics of various psychiatric disorders such as OCD, depression, autism, and schizophrenia. .. We have found that 18: 0/22: 6-PA produced by DGK ⁇ binds to Praja-1 and activates it, destabilizing SERT.
  • Non-Patent Document 12 Since DGK ⁇ has been found to bind to SERT and Praja-1 in the brain [Non-Patent Document 12, Non-Patent Document 13], in the present invention, by 18: 0/22: 6-PA produced by DGK ⁇ . A new mechanism for controlling SERT protein levels by activated Praja-1 can be proposed (Fig. 4). In addition, the present invention provides new insights into SERT level control and pathophysiology and therapeutic strategies for OCD, depression, autism, and schizophrenia.
  • 1-stearoyl-2-docosahexaenoyl (18: 0/22: 6) -phosphatidic acid (PA) [18: 0/22: 6-PA] is a serotonin transporter in the brain (serotonin). It is based on interacting with and activating Praja-1 [E3 ubiquitin-protein ligase] acting on transporter: SERT).
  • Drugs that promote squeezing include drugs such as preventive and therapeutic agents for the control of SERT, obsessive-compulsive disorder (OCD), major depressive disorder, autism, schizophrenia, and Alzheimer-type cognition.
  • OCD obsessive-compulsive disorder
  • the present invention has been completed by finding that it is excellent as a medicine for the prevention and treatment of illness and anxiety, depression, obsessive-compulsive disorder, and impulsive psychiatric disorder.
  • the present invention relates to serotonin transporter-related psychiatric disorders (depression, obsessive-compulsive disorder, schizophrenia, Alzheimer-type dementia, etc.) caused by those selected from docosahexaenoic acid and phosphatidic acid containing a docosahexaenoyl group or derivatives thereof.
  • serotonin transporter-related psychiatric disorders depression, obsessive-compulsive disorder, schizophrenia, Alzheimer-type dementia, etc.
  • serotonin transporter-related psychiatric disorders depression, obsessive-compulsive disorder, schizophrenia, Alzheimer-type dementia, etc.
  • serotonin transporter-related psychiatric disorders depression, obsessive-compulsive disorder, schizophrenia, Alzheimer-type dementia, etc.
  • the present invention provides the following: [1] docosahexaenoic acid (DHA) or its derivative, docosahexaenoyl group-containing phosphatidic acid or its derivative, and 1-stearoyl-2-docosahexaenoyl (18: 0/22: 6) -phosphatidic acid (PA)
  • DHA docosahexaenoic acid
  • PA 1-stearoyl-2-docosahexaenoyl
  • PA -phosphatidic acid
  • SERT activity inhibitor whose active ingredient is selected from the group consisting of salts thereof, selectively bound to Praja-1.
  • a Praja-1 activity enhancer that enhances its activity, or obsessive-compulsive disorder (OCD), obsessive-compulsive disorder, autism, schizophrenia, Alzheimer-type dementia, and anxiety, depression, obsessive-compulsive disorder, and urge.
  • OCD obsessive-compulsive disorder
  • Prophylactic / therapeutic agents for pathological symptoms selected from the group consisting of sexual psychiatric disorders, [2]
  • the SERT activity inhibitor is selected from the group consisting of DHA or a derivative thereof, docosahexaenoyl group-containing phosphatidic acid or a derivative thereof, or a salt thereof as an active ingredient.
  • the agent of [1] above, [4] Selected from the group consisting of docosahexaenoic acid (DHA) or its derivative, docosahexaenoyl group-containing phosphatidic acid or its derivative, and a drug compound that mimics 18: 0/22: 6-PA, or a salt thereof.
  • DHA docosahexaenoic acid
  • [5] Effective one selected from the group consisting of DHA or its derivatives, docosahexaenoyl group-containing phosphatidic acid or its derivatives, and chemical compounds mimic 18: 0/22: 6-PA, or salts thereof.
  • OCD obsessive-compulsive disorder
  • major depressive disorder autism
  • schizophrenia Alzheimer-type dementia
  • anxiety depression
  • obsessive-compulsive and impulsive psychiatric disorders.
  • the agent of [1] above which is a preventive / therapeutic agent for selected pathological symptoms.
  • OCD Obsessive-compulsive disorder
  • major depressive disorder autism
  • the present invention provides the following: [7] A selection from the group consisting of DHA or its derivatives, docosahexaenoyl group-containing phosphatidic acid or its derivatives, and chemical compounds mimic 18: 0/22: 6-PA, or salts thereof.
  • a method of suppressing the activity of SERT which is characterized by administration to a subject, a method of enhancing the activity of Praja-1 that selectively binds to Praja-1 and enhances its activity, or obsessive-compulsive disorder (OCD), major depression.
  • pathological symptoms selected from the group consisting of pathological disorders, autism, schizophrenia, Alzheimer-type dementia, and anxiety, depression, obsessive-compulsive, and impulsive psychiatric disorders.
  • the method of [7] above which comprises administering 18: 0/22: 6-PA.
  • the method of [7] above which comprises administering to a subject and suppressing the activity of SERT.
  • OCD obsessive-compulsive disorder
  • major depressive disorder autism
  • schizophrenia Alzheimer-type dementia
  • anxiety depression
  • obsessive-compulsive and impulsive psychiatric disorders.
  • the method of the above-mentioned [7] which comprises preventing / treating a pathological symptom selected from the above group.
  • the present invention provides the following.
  • [12] By measuring the activity selected from the group consisting of the interaction activity with Praja-1, the binding activity with Praja-1, and the enhancing or promoting activity of Praja-1 with respect to the drug candidate substance.
  • a screening method which comprises selecting and / or determining an active substance.
  • the screening method according to the above [12] which comprises analyzing the reaction of a drug candidate substance to the fusion protein AcGFP-Praja-1 or GST-Praja-1.
  • the above-mentioned [12] wherein the drug candidate substance is selected and / or determined by measuring the activity of enhancing the E3 ubiquitin-protein ligase activity of Praja-1.
  • Screening method [15] Select from the group consisting of obsessive-compulsive disorder (OCD), major depressive disorder, autism, schizophrenia, Alzheimer-type dementia, and anxiety, depression, obsessive-compulsive, and impulsive psychiatric disorders.
  • OCD obsessive-compulsive disorder
  • the activity selected from the group consisting of the interaction activity with Praja-1, the binding activity with Praja-1, and the enhancing or promoting activity of Praja-1 was measured, and the activity thereof was measured.
  • a screening kit comprising a Praja-1 protein or a tagged Praja-1 protein for selecting and / or determining what to have.
  • Docosahexaenoic acid (DHA) or a derivative thereof according to the present invention docosahexaenoyl group-containing phosphatidic acid or a derivative thereof, and 1-stearoyl-2-docosahexaenoyl (18: 0/22: 6) -phosphatidic acid (PA).
  • a drug compound that mimics [18: 0/22: 6-PA], or one that has an active ingredient selected from the group consisting of salts thereof, is a SERT activity inhibitor, selectively Praja-1.
  • Praja-1 activity enhancer that binds to and enhances its activity, or obsessive-compulsive disorder (OCD), obsessive-compulsive disorder, autism, schizophrenia, Alzheimer-type dementia, and anxiety, depression, obsessive-compulsive disorder. It is a prophylactic / therapeutic agent for pathological symptoms selected from the group consisting of sexual and impulsive psychiatric disorders, and controls serotonin / 5-hydroxytryptamine (5-HT) transporter (SERT), for example, SERT.
  • SERT serotonin / 5-hydroxytryptamine
  • the agent of the present invention can be expected to have the advantage of being highly safe as a pharmaceutical product or food.
  • various illnesses / diseases or pathological conditions can be treated as SERT.
  • Prevention and treatment of (related) are also provided.
  • an efficient screening method for active substances and compounds can be used, and useful active substances and active compounds can be analyzed and determined. Since 18: 0/22: 6-PA according to the present invention works through an action pathway different from SSRI and reduces SERT itself, a stronger action effect can be expected. It may also lead to increased drug choices for SERT-related psychiatric disorders.
  • COS-7 cells were transfected with AcGFP-Praja-1. After 48h incubation, cells are collected and cell lysates are placed in PC liposomes, 18: 1/18: 1-PS liposomes, 18: 0/22: 6-PG liposomes, 16: 0/18: 1-PA liposomes, 18: Incubate with 1/18: 1-PA liposomes, 18: 0/18: 0-PA liposomes, 18: 0/20: 4-PA liposomes, and 18: 0/22: 6-PA liposomes and then by ultracentrifugation.
  • DGK ⁇ selectively phosphorylates 18: 0/22: 6-DG to produce 18: 0/22: 6-PA.
  • 18: 0/22: 6-PA selectively bound to Praja-1 and enhanced its E3 ubiquitin-protein ligase activity.
  • Synaptic cleft indicates synaptic cleft
  • N-Terminal indicates N-terminal
  • C-Terminal indicates C-terminal
  • Proteasome indicates proteasome
  • Degradation indicates degradation
  • Presynapse indicates presynapse.
  • the present invention relates to docosahexaenoic acid (DHA) or a derivative thereof, docosahexaenoyl group-containing phosphatidic acid or a derivative thereof, and 1-stearoyl-2-docosahexaenoyl (18: 0/22: 6) -phosphatidic acid (PA).
  • DHA docosahexaenoic acid
  • PA 1-stearoyl-2-docosahexaenoyl
  • PA 1-stearoyl-2-docosahexaenoyl
  • the present invention was selected from the group consisting of A) DHA or its derivatives, docosahexaenoyl group-containing phosphatidic acid or its derivatives, and chemical compounds mimic 18: 0/22: 6-PA, or salts thereof.
  • DHA docosahexaenoic acid
  • ⁇ -3 fatty acid is also called all-cis-4,7,10,13,16,19-docosahexaenoic acid.
  • Examples of the DHA derivative include an ester derivative, an amide derivative, and a DHA derivative known to those skilled in the art, which are hydrolyzed in vivo to give DHA, and further used in vivo. Those that donate a possible docosahexaenoyl group may also be included.
  • DHA docosahexaenoyl group-containing phosphatidic acids, such as 18: 0/22: 6-PA, reduce the amount of serotonin transporter, 18: 0/22: 6-PA.
  • DHA can be ingested as a functional food, and the present invention may include those that can be ingested in the form of food as the above-mentioned preventive / therapeutic agent.
  • docosahexaenoyl group-containing phosphatidic acid or its derivative is, for example, the following general formula.
  • R 3 is a hydrogen atom, a group known to those skilled in the art of phospholipid chemistry, Examples thereof include groups that are hydrolyzed in vivo, such as choline residues, ethanolamine residues, serine residues, inositol residues, and glycerol residues, and groups that can be formed by in vivo metabolism.
  • the bond between the fatty acid and the glycerol skeleton is shown to be an ester bond [R- (CO) -OC], and the derivative thereof is the saturated aliphatic alkyl group or unsaturated fat described above.
  • the bond between a hydrocarbon residue including a group alkyl group and a glycerol skeleton is not only an ester bond but also an ether bond (ROC).
  • ROC ether bond
  • saturated fatty acids examples include formic acid, acetic acid, propionic acid, butyric acid (4: 0), valeric acid (5: 0), caproic acid (6: 0), enanthic acid (7: 0), and capric acid (8: 0).
  • Examples of the unsaturated fatty acid include ⁇ -3 fatty acid, ⁇ -6 fatty acid, ⁇ -7 fatty acid, ⁇ -9 fatty acid, and ⁇ -10 fatty acid.
  • Examples of the ⁇ -3 fatty acid include cis-7,10,13-hexadecatrienoic acid (16: 3), ⁇ -linolenic acid (ALA or 18: 3), stearidonic acid (STD or 18: 4), and the like.
  • ETE or 20: 3 Eicosatetraenoic acid
  • ETA or 20: 4 Eicosapentaenoic acid
  • EPA or 20: 5 Eicosapentaenoic acid
  • DPA or 22: 5 Eicosapentaenoic acid
  • DHA or 22: 6 Eicosahexaenoic acid
  • tetracosapentaenoic acid 24: 5
  • tetracosahexaenoic acid 24: 6 and the like.
  • Examples of the ⁇ -6 fatty acid include linoleic acid (18: 2), ⁇ -linolenic acid (18: 3), eikosazienoic acid (20: 2), dihomo- ⁇ -linolenic acid (20: 3), and arachidonic acid. (20: 4), docosadienoic acid (22: 2), docosatetraenoic acid (22: 4), docosapentaenoic acid (22: 5), calendic acid (18: 3) and the like.
  • Examples of the omega-7 fatty acid include palmitoleic acid (16: 1), vaccenic acid (18: 1), paullinic acid (20: 1) and the like.
  • Examples of the ⁇ -9 fatty acid include oleic acid (18: 1), elaidic acid (18: 1), eicosenoic acid (20: 1), mead acid (20: 3), erucic acid (22: 1), and the like. Examples include erucic acid (24: 1). Examples of the ⁇ -10 fatty acid include sapienic acid (16: 1).
  • O)- is a docosahexaenoyl group
  • R 3 is a hydrogen atom.
  • DHA docosahexaenoic acid
  • the docosahexaenoyl group at the 2-position is mostly derived from DHA ingested as food (ie, DHA is 18: 0 / Since it can be regarded as a precursor of 22: 6-PA), the prescribed medicinal effect can be obtained more effectively by ingesting DHA or administering DHA.
  • the above-mentioned DHA or a derivative thereof, and docosahexaenoyl group-containing phosphatidic acid or a derivative thereof can be synthesized by applying a chemical synthesis method.
  • these compounds can also be isolated and purified from in vivo metabolites when administered to mammals using conventional biochemical and chemical methods.
  • the chemical compounds that mimic 18: 0/22: 6-PA are not only those designed based on the chemical structural formula of 18: 0/22: 6-PA, but also their three-dimensional three-dimensional structural arrangement. It may be designed with reference to it, or it may be designed with reference to dynamic structural changes.
  • the drug compound that mimics the above 18: 0/22: 6-PA uses AI by analyzing the structural data of the Praja-1 protein and considering dynamic changes as necessary based on it. It may be obtained by the design.
  • the 1-position may be a shorter carbon chain corresponding to a fatty acid such as acetic acid, or the ester bond between the fatty acid and glycerol may be replaced with an ether bond.
  • the compound can be synthesized by applying a chemical synthesis method, or can be obtained by applying a biochemical method.
  • suitable salts include pharmaceutically acceptable salts such as salts with inorganic or organic bases, neutral, basic or acidic amino acids. Salt and the like.
  • salts with inorganic bases include alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium, and salts with aluminum and ammonium.
  • salt with an organic base include salts with trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N-dibenzylethylenediamine and the like.
  • salts with neutral amino acids include, for example, salts with glycine, valine, leucine and the like
  • suitable examples of salts with basic amino acids include, for example, salts with arginine, lysine, ornithine and the like.
  • the salt with an acidic amino acid for example, a salt with aspartic acid, glutamic acid and the like can be mentioned.
  • the drug containing the active ingredient of the present invention is an agent for controlling SERT, for example, an agent for controlling serotoninergic system through control of degradation and / or promotion of degradation of SERT, obsessive-compulsive disorder, depression. , Autism, and as a prophylactic / therapeutic agent for serotonin / SERT-related diseases such as schizophrenia.
  • the drugs containing the active ingredient of the present invention include SERT activity inhibitors, Praja-1 activity enhancers, and / or obsessive-compulsive disorder (OCD), major depressive disorder, autism, and schizophrenia.
  • Alzheimer-type dementia and useful as a prophylactic / therapeutic agent for pathological symptoms selected from the group consisting of anxiety, depression, obsessive-compulsive, and impulsive psychiatric disorders.
  • Disorders or disorders associated with the anxiety include neurosis, generalized anxiety disorder, social anxiety disorder, panic disorder, hyperactivity disorder, attention deficit disorder, personality disorder, bipolar disorder and the like.
  • the active ingredient of the present invention has sufficiently low toxicity and can be safely used as a pharmaceutical product.
  • the drug containing the active ingredient of the present invention is administered alone or in combination with other drugs as described later, preferably in the form of a preparation containing a pharmaceutically acceptable additive.
  • a preparation containing a pharmaceutically acceptable additive As the route of administration, oral, transdermal and injectable routes are adopted.
  • preparations of the above drugs external preparations (transdermal preparations, ointments, etc.), suppositories (rectal suppositories, vaginal suppositories, etc.), pellets, nasal preparations, inhalants (nebulizers, etc.), eye drops, liposomes. Examples include pharmaceutical products.
  • an ingredient selected from known preparation additives can be appropriately used regardless of the administration route.
  • Specific known pharmaceutical additives include, for example, (1) Handbook of Pharmaceutical Additives, Maruzen Co., Ltd., (1989), (2) Encyclopedia of Pharmaceutical Additives, 1st Edition, Yakuji Nippo Co., Ltd. (1994). , (3) Supplement to the Encyclopedia of Pharmaceutical Additives, 1st Edition, Yakuji Nippo Co., Ltd. (1995) and (4) Pharmaceutics, 5th Revised Edition, Nanedo Co., Ltd. (1997) Therefore, it can be appropriately selected according to the administration route and the intended use of the drug.
  • the additive may be any pharmaceutical component that can constitute an oral preparation and that can achieve the object of the present invention, but is usually an excipient or a binder.
  • Known pharmaceutical ingredients such as agents, disintegrants, lubricants, and coating agents (including taste masking) are selected.
  • Specific oral preparations include tablets (including sublingual tablets and orally disintegrating tablets), capsules (including soft capsules and microcapsules), granules, fine granules, powders, troches, syrups, and liposomes. And so on.
  • the oral preparation is a preparation in which the release of active drugs such as DHA and 18: 0/22: 6-PA, which are active ingredients, is controlled in the body by using known preparation ingredients (eg, immediate release). Formulations, sustained release preparations) are also included.
  • a pharmaceutical component that can constitute an aqueous injection or a non-aqueous injection is used, and usually, a solubilizing agent, a solubilizing agent, a suspending agent, an isotonicizing agent, and a buffering agent.
  • Stabilizers, preservatives and other known pharmaceutical ingredients are used, but they may also be known pharmaceutical ingredients constituting powder injections for use by dissolving or suspending them at the time of administration.
  • the pharmaceutical components of the aqueous injection include, for example, distilled water for injection, isotonic sterilized salt solution (1 sodium or 2 sodium phosphate, sodium chloride, potassium chloride, calcium chloride or magnesium chloride, etc., or such.
  • Examples of the pharmaceutical components of the non-aqueous injection include vegetable oils such as olive oil, sesame oil, cottonseed oil, and corn oil, propylene glycol, macrogold, and tricapryrin, which are dissolved in these. Manufactured by suspension or emulsification. Specific examples of the injection include a subcutaneous injection, an intravenous injection, an intramuscular injection, an intraperitoneal injection, a drip infusion, and a liposome.
  • the usage of the active drug or active compound such as DHA, 18: 0/22: 6-PA for preparing a drug (pharmaceutical or pharmaceutical composition) having the above-mentioned activity containing the active ingredient of the present invention is also possible. Provided.
  • the effective dose of the drug of the present invention varies depending on the age, weight, symptom of illness, presence or absence of complications, etc. of the patient to be administered, and is appropriately adjusted. However, in the case of oral administration, it is usually 0.1 mg to 3,000. Administer about mg / day, or in the case of injection, about 0.1 mg to 1,000 mg / day.
  • the drug as the active ingredient of the present invention can also be used in combination with one or more other drugs that do not adversely affect the effect for the purpose of enhancing the action, lowering the dose, reducing side effects and the like.
  • the concomitant drug that can be combined may be a small molecule compound, a polypeptide, an antibody, a vaccine or the like.
  • the administration form in combination with the concomitant drug is not particularly limited, and the drug may be simply used in combination.
  • both are formulated simultaneously and administered as a single formulation, both are formulated separately and administered simultaneously or at different time intervals on the same route of administration, and both are formulated separately and administered simultaneously or on different routes of administration.
  • the administration etc. may be mentioned.
  • the concomitant drug can be selected from, for example, a SERT inhibitor (SSRI), and the SERT inhibitor can be, for example, Fluvoxamine, fluoxetine, paroxetine, sertraline, citalopram, escitalopram and the like can be mentioned.
  • SERT inhibitor can be, for example, Fluvoxamine, fluoxetine, paroxetine, sertraline, citalopram, escitalopram and the like can be mentioned.
  • the present invention also provides a preventive / therapeutic method for diseases and the like using the active ingredient drug (compound), for example, by administering an effective amount of the 18: 0/22: 6-PA to a subject.
  • the preventive / therapeutic method can be implemented.
  • This prevention / treatment method can also be performed while monitoring the health condition and progress of symptoms of the subject.
  • the monitor may be performed at regular or irregular time intervals, or may be performed regularly. In a typical case, it is performed while monitoring blood HDL levels.
  • the administration may be performed at regular or irregular time intervals, or may be performed regularly.
  • prevention / treatment may refer to prevention and / or treatment, and includes cases where it means prevention and treatment, cases where it means prevention, and cases where it means treatment. is doing.
  • OCD obsessive-compulsive disorder
  • major depressive disorder autism
  • schizophrenia Alzheimer-type dementia
  • anxiety depression
  • Foods that have the preventive / therapeutic effect of the selected pathological symptoms include DHA or its derivatives, docosahexaenoyl group-containing phosphatidic acid or its derivatives, and 18: 0/22: 6-PA as its active ingredients. It contains a chemical compound selected from the group consisting of chemical compounds or salts thereof. In particular, foods containing DHA can be mentioned.
  • DHA is a fish oil obtained from fish such as herring, mackerel, sardines, tuna, bonito, saury, and yellowtail, and its content is known to be high. It is also known to be produced by microorganisms such as Euglena, which can also be obtained from fermentation products of such microorganisms.
  • the oil product containing DHA those having various DHA purity and DHA composition are commercially available or known, and those having the intended effect of the present invention can be selected and used from these products.
  • the food of the present invention may be provided as a composition in various forms, and the food itself containing a functional food, an additive used in producing a food containing various processed foods and functional foods, It can be in the form of an animal feed itself, an additive used in producing an animal feed, or the like. These various forms can be produced by a commonly used method.
  • the food to which the present invention is applied includes all foods including beverages, and in addition to general processed foods including so-called health foods, foods for specified health use and nutrition specified in the Health Function Food System of the Consumer Affairs Agency of Japan. Includes health functional foods such as functional foods, supplements, etc., as well as health functional foods such as specified health foods and nutritionally functional foods, supplements, etc. that are supported in countries other than Japan, and also includes feeds fed to animals. do.
  • Foods such as the functional foods of the present invention can take a solid, semi-solid or liquid form, for example, oral liquids (including drinks), biscuits, confectionery, candy, tablets (tablet confectionery, etc.). (Including), granules (including granular confectionery, etc.), powders (including powdered beverages, powdered confectionery, etc.), capsules, jelly and the like.
  • the food product form of the present invention includes, for example, beverages (soft beverages, tea beverages, coffee beverages, dairy beverages, fruit juice beverages, carbonated beverages, nutritional drinks, powdered beverages, alcoholic beverages, non-alcoholic beverages, sports drinks, etc.
  • Soybean processed foods, breads, noodles, rice, gel-like foods jelly drinks, jelly, bavarois, pudding, mousse, gummy candy, etc.
  • sweets variant snacks, baked goods, cakes, chocolate , Gum, candy, tablets, etc.
  • soups dairy products, frozen foods, processed marine products (fish sausage, kamaboko, chikuwa, hampen, etc.), processed livestock products (hamburger, ham, sausage, wiener, cheese, butter, yogurt) , Raw cream, margarine, fermented milk, etc.), instant foods, supplements, capsules, cereals, other processed foods, seasonings and their ingredients.
  • the food can further contain other necessary raw materials or additives as appropriate, as long as the effects of the present invention are not impaired. Examples of other raw materials or additives include fruit juices, sweeteners, acidulants, vitamins, amino acids, minerals, proteins, thickeners, flavors, pigments and the like.
  • Instructions can be attached to the product. This description can be provided by attaching the instruction manual prepared separately from the product to the product package, or by printing the instruction manual on the product itself or on the packaging of the product (including the inner bag that wraps the divided product). ..
  • information regarding the content of the active ingredient such as DHA of the product, the total intake amount of the active ingredient such as DHA within the ingestion time, the period of continuous ingestion, and the like can be described.
  • the products can be classified according to the amount to be ingested within the ingestion time range, and the required amount of the divided products can be stored in the product package.
  • the activity selected from the group consisting of the activity of interacting with Praja-1, the activity of binding to Praja-1, and the activity of enhancing or promoting the activity of Praja-1 is measured.
  • the Praja-1 protein is preferably used.
  • the Praja-1 used for screening may be in vivo or in vitro.
  • the fusion protein Praja-1 produced by applying genetic engineering technology is used.
  • a fusion protein or the like with a tag that binds to the ligand of.
  • the tag of the Praja-1 protein any suitable marker can be appropriately selected and applied as long as it is a marker that can be detected.
  • Typical examples of the fusion protein include AcGFP-Praja-1, GST-Praja-1 and the like. Detection can also be performed using an antibody (including a monoclonal antibody and a fragment thereof) that specifically recognizes the fusion tag. Expression and purification of such fusion polypeptides or fusion proteins can be performed using commercially available kits suitable for them and can also be performed according to the protocol disclosed by the kit manufacturer or kit distributor.
  • Performing this screening can be performed using standard techniques that are well known and conventional to those skilled in the art.
  • SERT activity inhibitor Praja-1 activity enhancer that selectively binds to Praja-1 and enhances its activity
  • OCD obsessive-compulsive disorder
  • Efficient active substances and active compounds useful as prophylactic / therapeutic agents for pathological symptoms selected from the group consisting of obsessive-compulsive disorder, anxiety, depression, obsessive-compulsive disorder, and impulsive psychiatric disorders. It is possible to perform analysis with priority and priority and select it.
  • Examples of the above-mentioned pharmaceutical candidate substances include compounds or compositions obtained by chemical synthesis, naturally derived compounds or compositions thereof, metabolites, fermentation products, phospholipids, lipids, peptides, proteins, sugar chains and other biopolymer compounds. , Or a mixture thereof.
  • the test substance in the screening method of the present invention is not particularly limited. For example, it may be a compound such as a small molecule compound or a high molecular compound.
  • this screening involves (i) a system containing the test sample and the Praja-1 protein, and (ii) a system containing the Praja-1 protein but not adding the test sample.
  • the above reaction is allowed to proceed, the presence or absence of binding to the Praja-1 protein, the amount of binding, etc. are measured, and either of these is measured in (i) and (ii).
  • a suitable detection substrate may be present in the screening system so as to be convenient for measurement.
  • the substrate may be any substrate as long as it can be effectively used for measurement.
  • a compound known as a known substrate can be selected and used, and a synthesized compound or the like can be preferably used.
  • the substrate can be used as it is, but preferably a substrate labeled with a fluorescent, enzyme or radioactive substance such as fluoressein can be used.
  • the Praja-1 protein may be one in which the tagged Praja-1 protein is expressed in animal cells, for example, COS cells, CH0 cells, human-derived cell lines, iPS cells, and the like.
  • animal cells for example, COS cells, CH0 cells, human-derived cell lines, iPS cells, and the like.
  • an appropriate selection marker can be used to obtain a gene in which the target protein is expressed at a higher level, and the target protein is expressed. It can also be cultivated under possible conditions to produce and accumulate the desired product.
  • the transformant cells can be cultured in a medium widely used in the art.
  • a MEM medium containing about 5 to about 20% fetal bovine serum, PRMI1640 medium, DMEM medium, or the like may be used as the medium.
  • the pH is preferably about 6 to about 8.
  • Culturing is usually carried out at about 30 to about 40 ° C. for about 15 to about 72 hours, and aeration and stirring are added as necessary.
  • Transformed animal cells expressing a predetermined protein can be used as they are, but they can also be used as cell homodunates thereof, or a predetermined gene product protein can be isolated and used.
  • the cells When extracting from the above cultured cells, after culturing, the cells are collected by a known method, suspended in an appropriate buffer solution, destroyed by ultrasonic waves, lysozyme and / or freeze-thaw, and then centrifuged. A method of obtaining a crude extract by filtration or the like can be appropriately used.
  • a protein denaturing agent such as urea or guanidine hydrochloride, or a surfactant such as Triton X-100 (trade name) or Twien-20 (trade name) may be added to the buffer solution.
  • the target protein can be purified by appropriately combining the separation / purification methods known per se.
  • This screening can be carried out according to a usual method for measuring binding activity or enzyme activity, and can be carried out with reference to, for example, a method known in the art.
  • various labels, buffer systems, other appropriate reagents, etc. can be used, and the operation can be performed according to the operation described therein.
  • the protein to be used can be treated with an activator, or its precursor or latent form can be converted into an active form in advance.
  • the measurement is usually carried out in a buffer solution such as Tris-HCl buffer solution or phosphate buffer solution which does not adversely affect the reaction, for example, at pH of about 4 to about 10 (preferably pH of about 6 to about 8). It can be carried out.
  • the screening kit of the present invention contains a Praja-1 protein or a tagged Praja-1 protein.
  • the screening kit of the present invention can screen a substance or compound having an activity selected from the group consisting of an interaction activity with Praja-1, a binding activity with Praja-1, and an enhancing or promoting activity of Praja-1. ..
  • the screening kit also includes obsessive-compulsive disorder (OCD), major depressive disorder, autism, schizophrenia, Alzheimer-type dementia, and anxiety, depression, obsessive-compulsive, and impulsive psychiatric disorders. It is possible to screen substances or compounds having a preventive / therapeutic effect on pathological symptoms selected from the group.
  • the screening kit may include a buffer solution for a reaction, a reagent, or the like as described above as a set so that it is convenient to perform the required measurement.
  • a buffer solution for a reaction for a reaction, a reagent, or the like as described above as a set so that it is convenient to perform the required measurement.
  • the present invention will be specifically described with reference to examples, but the present invention is provided merely for the purpose of explaining the present invention and for reference in a specific embodiment thereof. These examples are for explaining specific specific embodiments of the present invention, but do not represent limiting or limiting the scope of the invention disclosed in the present application. It should be understood that in the present invention, various embodiments based on the ideas of the present specification are possible. All embodiments have been or can be carried out using standard techniques, except as described in detail elsewhere, which are well known and conventional to those of skill in the art. ..
  • Mouse Praja-1 (NCBI accession no. XM_011247544) cDNA was amplified from mouse brain cDNA and inserted into the EcoRI / SalI site of the pAcGFP vector [Non-Patent Document 13].
  • GST glutathione S-transferase
  • the Praja-1 cDNA should be used in pSF-CMV-Puro-NH2-GST-TEV (Oxford Genetics, Oxford, UK) EcoRI /. I also ligated to the XhoI site.
  • COS-7 cells are Dulbecco's modified Eagle's medium (D-MEM) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA) and 100 U / mL penicillin + 100 ⁇ g / mL streptomycin; Wako Pure Chemical Industries, Osaka, Japan) maintained the temperature at 37 ° C in an atmosphere containing 5% CO 2 .
  • D-MEM Dulbecco's modified Eagle's medium
  • penicillin + 100 ⁇ g / mL streptomycin Wako Pure Chemical Industries, Osaka, Japan
  • pAcGFP-Praja-1 or pSF-CMV-Puro-NH2-GST-TEV-Praja-1 was transfected as shown in the legend in the figure in the kit manual.
  • the plasmid was transiently transfected with PolyFect (Qiagen, Hilden, Germany) according to the kit manufacturer's instructions.
  • Mouse cerebral cortex was homogenized in ice-cold Lysis buffer (50 mM HEPES, pH7.2, 150 mM NaCl, 5 mM MgCl 2 , cOmplete TM EDTA-free protease inhibitor) and then 1000 g at 4 ° C. Centrifuge for 5 minutes. Total lipids were extracted from mouse brain (cerebral cortex) according to Bligh and Dyer's method [Non-Patent Document 15]. 2 ml of methanol and 1 mL of chloroform were added to 700 ⁇ L of the sample.
  • Non-Patent Document 16 For PA analysis, 100 ⁇ L of 3M HCl was added to the sample to improve the recovery ratio of acidic phospholipids [Non-Patent Document 16].
  • the lipid-containing solvent was dried under N 2 gas and the extracted lipid was reconstituted in 100 ⁇ L chloroform / methanol (2: 1, v / v).
  • GST-Praja-1 was expressed by COS-7 cells. After cytolysis and centrifugation, affinity-chromatography with glutathione sepharose 4B (GE Healthcare, Chicago, IL, USA) was performed to purify GST fusion ⁇ -Syn-N. The column was washed with lysis buffer and the bound protein was eluted with buffer containing 50 mM Tris / HCl, 150 mM NaCl, and 10 mM reduced glutathione. For the lipid overlay assay, the purified protein was dialyzed against HEPES buffer (25 mM HEPES, pH 7.4, 100 mM NaCl). Protein concentration was measured by the Bicinchoninic Acid Protein Assay Kit (Thermo Fisher Scientific).
  • lipid binding properties of the Praja-1 protein were determined using the following lipid mixture: Control liposomes [Cholesterol (30 mol% (Wako Pure Chemical Industries, Ltd.)) and phosphatidylcholine (PC, from egg yolk (Avanti Polar Lipids, Alabaster, AL, USA)) (70 mol%)], phosphatidylserine (PS) liposomes [ Chol (30 mol%), PC (from egg yolk) (60 mol%), and 18: 1/18: 1-PS (10 mol% (Avanti Polar Lipids))], phosphatidylglycerol (PG) liposomes [Chol (30 mol%), PC (from egg yolk) (60 mol%), and 18: 0/22: 6-PG (10 mol% (Avanti Polar Lipids))], and PA liposomes [Chol (30 mol%), PC (from egg yolk) (60 mol%) , And each PA species (10 mol%)
  • PA species include 16: 0/18: 1-PA (Avanti Polar Lipids), 18: 1/18: 1-PA (Avanti Polar Lipids), 18: 0/18: 0-PA (Avanti Polar Lipids), 18: 0/20: 4-PA (Avanti Polar Lipids) and 18: 0/22: 6-PA (Avanti Polar Lipids) were used.
  • the combined dry lipid mixture was hydrated with HEPES buffer (25 mM HEPES, pH 7.4, 100 mM NaCl, 1 mM dithiothreitol) at 95 ° C for 45 min and 15 min during hydration. Vortexed once for 1 min. The liposomes were then thawed for 5 cycles (3 min -196 ° C, 3 min 95 ° C).
  • Liposomal formation [Non-Patent Document 21] was induced by sonication at 95 ° C using Branson Sonifier 450 (Branson Ultrasonics Corporation, Danbury, CT, USA).
  • COS-7 cells were transfected with pAcGFP-Praja-1. After incubation for 48 hours, cells were collected with HEPES buffer and the disrupted product (0.3 mg) was incubated with PA-containing liposomes or control liposomes at 4 ° C for 30 min. The sample was ultracentrifuged at 200000 g at 4 ° C for 1 h. The precipitate was dissolved in HEPES buffer. Then, using a CS100GXII centrifuge and an S100-AT4 angle rotor (Hitachi Koki, Tokyo, Japan), the sample was centrifuged at 200,000 g at 4 ° C for 1 h. The precipitate was dissolved in HEPES buffer.
  • E3 Ubiquitin-Protein Ligase Activity Assay Purified GST-Praja-1 fusion protein was added to PC liposomes, 18: 0/22: 6-PG liposomes, 18: 1/18: 1-PA liposomes, 18: 0/18: 0-PA liposomes, and 18 :. Incubated with 0/22: 6-PA liposomes.
  • the E3 ligase self-ubiquitination assay kit (BML-UW0970; Enzo Life Sciences, NY, NY, USA) was used to analyze E3 ubiquitin-protein ligase activity. The assay was performed according to the manufacturer's protocol.
  • each reaction (25 ⁇ L final volume) is 1.25 ⁇ L 20 ⁇ E1, 1.25 ⁇ L 20 ⁇ E2, 2.5 ⁇ L 10 ⁇ Ub E3 ligase buffer, 2.5 ⁇ L 10 ⁇ ubiquitin, 0.25 ⁇ L 100 mM. It was made to contain DTT and 1.25 ⁇ L of Mg-ATP.
  • the reaction mixture was incubated at 37 ° C for 1 h and SDS / PAGE using anti-ubiquitin antibody (included in the kit) and horseradish peroxyda-zeconjugated goat anti-mouse IgG (Bethyl Laboratories, Montgomery, AL, USA). / Analyzed by Western blotting.
  • LC-MS / MS analysis shows that 40: 6-PA contains docosahexaenoic acid (DHA, 22: 6, ⁇ -3) at the sn-2 position 18: 0/22: 6-PA (94.6). %) Shown that it is mainly (Table 1 and Table 2).
  • Tables 1 and 2 below show the results of identifying acyl species of each PA molecular species in the mouse cerebral cortex using LC-MS / MS.
  • Tables 3 and 4 below show the results of identifying acyl species of each DG molecular species in the mouse cerebral cortex using LC-MS / MS.
  • Praja-1 did not precipitate with mock control (without liposomes) (Fig. 2A).
  • Liposomes containing PC (neutral phospholipid) alone as a background control showed moderate precipitation ( ⁇ 50%) of Praja-1 (FIGS. 2A, B).
  • PS (acidic phospholipid) liposomes did not precipitate Praja-1 more strongly than PC-controlled liposomes.
  • 18: 0/22: 6-PG liposomes as acidic phospholipid controls containing the same fatty acid moieties as 18: 0/22: 6-PA also have almost the same Praja-1 binding activity as background controls. showed.
  • nearly 100% of Praja-1 co-precipitated with 18: 0/22: 6-PA liposomes (Fig. 2A, B).
  • AcGFP alone as a negative control was not detectable in precipitated 18: 0/22: 6-PA liposomes (Fig. 2A).
  • Fig. 2A, B the Praja-1 binding activity of other PA species was also determined. Unlike 18: 0/22: 6-PA liposomes, 16: 0/18: 1-PA liposomes, 18: 0/18: 0-PA liposomes, and 18: 1/18: 1-PA liposomes are PCs. Only the co-precipitation ability, which is almost the same as that of the control liposome, was shown (Fig. 2A, B). Liposomes containing 18: 0/20: 4-PA, also with polyunsaturated fatty acid (PUFA) arachidonic acid (20: 4, ⁇ -6), have stronger Praja-1 binding activity than PC control liposomes. Was not shown (Fig. 2A, B).
  • PUFA polyunsaturated fatty acid
  • Praja-1 is a liposome containing phospholipids, such as PS liposomes (acidic phospholipid control), 18: 0/22: 6-PG liposomes (acidic phospholipid control containing the same fatty acid moiety), and other PA species.
  • Non-Patent Document 16 Non-Patent Document 20, Non-Patent Document 22
  • DGK ⁇ -KO mouse cerebral cortex at 18: 0/22: 6 -It was demonstrated that PA and 18: 0/22: 6-DG were simultaneously decreasing and accumulating (Fig. 1).
  • DGK ⁇ utilizes 18: 0/22: 6-DG to generate 18: 0/22: 6-PA.
  • 18: 0/22: 6-PA selectively binds to Praja-1 and enhances the E3 ubiquitin-protein ligase activity of purified Praja-1. (Figs. 2 and 3).
  • Non-Patent Document 14 which ubiquitinates SERTs [Non-Patent Document 14]. It was clarified that Document 12, Non-Patent Document 13] and that Praja-1 induces SERT ubiquitination and degradation in a DGK activity-dependent manner [Non-Patent Document 13]. Therefore, the increase in 18: 0/22: 6-PA produced by DGK ⁇ directly enhances Praja-1 E3 ubiquitin-protein ligase activity, resulting in a decrease in SERT protein stability in the brain. Is possible (Fig. 4).
  • PI 4,5-bisphosphate which is made by PI turnover and consists primarily of 18: 0/20: 4-PI 4,5-diphosphate species
  • PI phosphatidylinositol
  • 6PA structurally simple lipid 18: 0/22: 6-PA was also Praja-1. It suggests that it plays an essential role in the central nervous system through the activation of E3 ubiquitin-protein ligase and the degradation of SERT.
  • DGK ⁇ produces 16: 0-containing PA species and 16: 1-containing PA species in C2C12 myoblasts in response to high glucose stimulation [Non-Patent Document 22]. .. However, the study of this example strongly suggested that DGK ⁇ produces 18: 0/22: 6-PA in the brain (Fig. 1). DGK ⁇ has no obvious DG species selectivity in vitro [Non-Patent Document 22]. Therefore, it can be said with some high possibility that DGK ⁇ utilizes different DG species pools in different cells and tissues.
  • DGK ⁇ is 30: 0 (14: 0/16: 0) -PA, 32: 0 (16: 0/16: 0) -PA, and 34: 0 (16: 0 / 18: 0)-PA was generated [Non-Patent Document 25]. Therefore, it is possible that different PA species are produced by DGK isozymes in different tissues and cells [Non-Patent Document 26]. Like DGK ⁇ , DGK ⁇ and DGK ⁇ showed no apparent DG species selectivity in vitro [Non-Patent Document 27, Non-Patent Document 28]. Therefore, they may also access different DG species pools.
  • Non-Patent Document 29 Since different DGK isozymes produce different PA species in different tissues and cells, it is speculated that these different PA species have their own unique targets. In fact, the present inventors have found that 18: 1/18: 1-PA selectively interacts with ⁇ -synuclein [Non-Patent Document 29, Non-Patent Document 30].
  • muscular creatine kinases are selectively 16: 0/16: 0-PA, 16: 0/18: 1-PA, 18: 1/18: 1-PA, and 18: 0/18: 0- Bound to PA [Non-Patent Document 31], L-lactic dehydrogenase A selectively 16: 0/16: 0-PA, 18: 0/18: 0-PA, 18: 0/20: 4-PA, And 18: 0/22: 6-bonded to PA.
  • DHA docosahexaenoic acid
  • ⁇ -3 PUFAs such as DHA have anxiolytic effects
  • DHA-containing PA selectively binds to and activates SERT (which is closely related to anxiety [Non-Patent Document 5]) E3 ubiquitin-protein ligase Praja-1. (Figs. 2 and 3). Therefore, it is possible that DHA incorporated into 18: 0/22: 6-PA, which enhances SERT degradation, could explain the anxiolytic effect of DHA. In addition to its free fatty acid form, it can be said with some high probability that DHA incorporated into PA also plays an important role in brain function.
  • Non-Patent Document 35 A female patient with DGK ⁇ gene disruption was recently investigated [Non-Patent Document 35]. Patients showed seizures and experienced self-stimulating behaviors such as pulling hair, tapping their feet, and fluttering their hands [Non-Patent Document 35]. These are observed in OCD and OC spectrum disorders [Non-Patent Document 36, Non-Patent Document 37]. Therefore, abnormalities in DGK ⁇ cause OCD-like psychiatric disorders in humans, meaning that DGK ⁇ is a key enzyme for maintaining mental health. However, the regulatory mechanism of DGK ⁇ activity / expression remains unknown.
  • PKC protein kinase C
  • DGK ⁇ phosphorus
  • PKC ⁇ can regulate the intracellular localization and activity of DGK ⁇ in the brain.
  • formal esters increased the expression level of DGK ⁇ [Non-Patent Document 43], suggesting that conventional PKC and Nobel PKC, including PKC ⁇ , regulate their expression.
  • SERT inhibitors have been used to treat depression or major depressive order and OCD [Non-Patent Documents 2-4], with excess SERT protein / activity in these. It means that it is closely related to the onset of the disorder. Moreover, SERT is associated with anxiety and autism [Non-Patent Document 5]. Hyperactivity of 5-HT is associated with negative signs of schizophrenia [Non-Patent Document 44]. In this regard, treatment with a potent 5-HT 2A receptor antagonist in combination with a weak dopamine D2 receptor antagonist is the first-line treatment for patients suffering from schizophrenia (negative symptoms) [Non-Patent Document 44]. ..
  • active substances and active compounds can be identified by using the interaction activity with Praja-1, the binding activity with Praja-1, the enhancing or promoting activity of Praja-1 as an index, and further.
  • Drugs with identified activity including DHA or derivatives thereof, docosahexaenoyl group-containing phosphatidic acid or derivatives thereof, drug compounds mimic 18: 0/22: 6-PA, or salts thereof, are SERTs.
  • Activity inhibitor Praja-1 activity enhancer that selectively binds to and enhances its activity, or obsessive-compulsive disorder (OCD), major depressive disorder, autism, schizophrenia, It can be used as a prophylactic / therapeutic agent for Alzheimer-type dementia and pathological symptoms selected from the group consisting of anxiety, depression, obsessive-compulsive, and impulsive psychiatric disorders, and is highly useful. It is clear that the present invention can be carried out other than those specifically described in the above description and examples. In view of the above teachings, many modifications and variations of the invention are possible, and thus they are also within the scope of the appended claims.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biomedical Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Neurology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Neurosurgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Psychiatry (AREA)
  • Food Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Pain & Pain Management (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biophysics (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Mycology (AREA)
  • Nutrition Science (AREA)
  • Polymers & Plastics (AREA)
  • Hospice & Palliative Care (AREA)
PCT/JP2021/032309 2020-09-04 2021-09-02 ドコサヘキサエン酸、ドコサヘキサエノイル基含有ホスファチジン酸、又はその誘導体によるセロトニントランスポーター関連精神疾患の予防・治療剤 Ceased WO2022050355A1 (ja)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2022546971A JPWO2022050355A1 (https=) 2020-09-04 2021-09-02

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2020-149427 2020-09-04
JP2020149427 2020-09-04

Publications (1)

Publication Number Publication Date
WO2022050355A1 true WO2022050355A1 (ja) 2022-03-10

Family

ID=80491124

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2021/032309 Ceased WO2022050355A1 (ja) 2020-09-04 2021-09-02 ドコサヘキサエン酸、ドコサヘキサエノイル基含有ホスファチジン酸、又はその誘導体によるセロトニントランスポーター関連精神疾患の予防・治療剤

Country Status (2)

Country Link
JP (1) JPWO2022050355A1 (https=)
WO (1) WO2022050355A1 (https=)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007509131A (ja) * 2003-10-22 2007-04-12 エンジィモテック リミテッド オメガ−３およびオメガ−６脂肪酸を含有するグリセロリン脂質
JP2007518764A (ja) * 2004-01-21 2007-07-12 ブルツツエーゼ，テイベリオ 中枢神経系障害の治療のための、選択されたｎ−３系脂肪酸の高度濃縮組成物の使用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007509131A (ja) * 2003-10-22 2007-04-12 エンジィモテック リミテッド オメガ−３およびオメガ−６脂肪酸を含有するグリセロリン脂質
JP2007518764A (ja) * 2004-01-21 2007-07-12 ブルツツエーゼ，テイベリオ 中枢神経系障害の治療のための、選択されたｎ−３系脂肪酸の高度濃縮組成物の使用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
NISHI DAISUKE J, YUTAKA MATSUOKA: "Evidence of omega-3 fatty acids against depression and PTSD", JPN J GENERAL HOSPITAL PSYCHIATRY, vol. 25, no. 3, 1 January 2013 (2013-01-01), pages 248 - 253, XP055906688 *
QIANG LU; CHIAKI MURAKAMI; YUKI MURAKAMI; FUMI HOSHINO; MAHO ASAMI; TAKAKO USUKI; HIROMICHI SAKAI; FUMIO SAKANE: "1‐Stearoyl‐2‐docosahexaenoyl‐phosphatidic acid interacts with and activates Praja‐1, the E3 ubiquitin ligase acting on the serotonin transporter in the brain", FEBS LETTERS, vol. 594, no. 11, 19 March 2020 (2020-03-19), NL , pages 1787 - 1796, XP071256985, ISSN: 0014-5793, DOI: 10.1002/1873-3468.13765 *
陸強 ほか, ジアシルグリセロールキナーゼδが産生する18:0/22:6-ホスファチジン酸を介してセロトニントランスポーターのユビキチン化を亢進しそのタンパク質量を制御する, 日本生化学会第９３回日本生化学会大会要旨, 01 September 2020, pp. 0881-0882, [P-089] *
陸強 ほか, ジアシルグリセロールキナーゼδが産生する18:0/22:6-ホスファチジン酸を介してセロトニントランスポーターのユビキチン化を亢進しそのタンパク質量を制御する, 脂質生化学研究, 01 May 2020, vol. 62, pp. 208-211, 2-23 *

Also Published As

Publication number Publication date
JPWO2022050355A1 (https=) 2022-03-10

Similar Documents

Publication Publication Date Title
He et al. Cellular uptake, metabolism and sensing of long-chain fatty acids
Qu et al. Lipid-induced S-palmitoylation as a vital regulator of cell signaling and disease development
Rådmark et al. Regulation of the activity of 5-lipoxygenase, a key enzyme in leukotriene biosynthesis
AU2018201800C1 (en) Lipid scavenging in Ras cancers
Cacabelos et al. Interplay between TDP-43 and docosahexaenoic acid-related processes in amyotrophic lateral sclerosis
Holler et al. Dietary choline supplementation in pregnant rats increases hippocampal phospholipase D activity of the offspring
Opreanu et al. Inhibition of cytokine signaling in human retinal endothelial cells through downregulation of sphingomyelinases by docosahexaenoic acid
Lu et al. 1‐Stearoyl‐2‐docosahexaenoyl‐phosphatidic acid interacts with and activates Praja‐1, the E3 ubiquitin ligase acting on the serotonin transporter in the brain
Zhang et al. Monoacylglycerol acyltransferase-2 is a tetrameric enzyme that selectively heterodimerizes with diacylglycerol acyltransferase-1
CN104321300A (zh) 脂肪酸的2-羟基衍生物的对映体
Kim et al. Dietary n-6 PUFA deprivation downregulates arachidonate but upregulates docosahexaenoate metabolizing enzymes in rat brain
Tsushima et al. Lysophosphatidylserine form DHA maybe the most effective as substrate for brain DHA accretion
Honzíková et al. Novel approaches for elongation of fish oils into Very-Long-Chain polyunsaturated fatty acids and their enzymatic interesterification into glycerolipids
López-Nicolás et al. Molecular mechanisms of PKCα localization and activation by arachidonic acid. The C2 domain also plays a role
WO2022050355A1 (ja) ドコサヘキサエン酸、ドコサヘキサエノイル基含有ホスファチジン酸、又はその誘導体によるセロトニントランスポーター関連精神疾患の予防・治療剤
Rajakumar et al. Dysregulation of ceramide metabolism causes phytoceramide-dependent induction of the unfolded protein response
Numagami et al. Distinct regions of Praja-1 E3 ubiquitin-protein ligase selectively bind to docosahexaenoic acid-containing phosphatidic acid and diacylglycerol kinase δ
US9512152B2 (en) Phospholipid compound containing unsaturated fatty acid derivative having cyclopropane ring
Levy et al. Polyisoprenyl phosphates: natural antiinflammatory lipid signals
JP2008260743A (ja) Ｎｑｏ１発現増強剤
EP4302780A1 (en) Composition for preventing or treating cerebrovascular accident
Basak et al. Maternal supply of both arachidonic and docosahexaenoic acids is required for optimal neurodevelopment. Nutrients. 2021; 13: 2061
Verdugo Effect of lipid supplementation on ruminal epithelial membrane fatty acid composition and short-chain fatty acid absorption
Virmani et al. Neuronal carnitine palmitoyl transferase 1c in the central nervous system: Current visions and perspectives
He et al. Allosteric activation of a cell-type-specific GPR120 inhibits amyloid pathology of Alzheimer’s disease

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21864408

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2022546971

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21864408

Country of ref document: EP

Kind code of ref document: A1