JP2008208091A - Curative agent and prophylactic agent for endoplasmic reticulum stress-related disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical composition for treating and/or preventing endoplasmic reticulum stress-related diseases in neurocytes. <P>SOLUTION: The pharmaceutical composition is provided, being effective for treating and/or preventing endoplasmic reticulum stress-related diseases without being accompanied by such a side effect as to cause neurocyte death by NSAIDs (nonsteroidal anti-inflammatory drugs) such as pranoprofen. Therefore, a curative agent and a prophylactic agent for endoplasmic reticulum stress-related diseases in neurocytes such as Alzheimer's disease, Parkinson's disease and cerebral ischemia can be provided. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to therapeutic agents for endoplasmic reticulum (ER) related diseases (eg, central nervous system (CNS) diseases such as Alzheimer's disease, Parkinson's disease, and cerebral ischemia).

  Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used as therapeutic agents in the treatment of fever, pain, and inflammation. On the other hand, in addition to these effects, it has recently been suggested that nonsteroidal anti-inflammatory drugs have other actions. As a result of clinical experiments, it has been shown that some non-steroidal anti-inflammatory drugs are effective in the treatment and treatment of cancer (Non-patent Document 1). Furthermore, epidemiological studies have shown that non-steroidal anti-inflammatory drugs are associated with reduced risk of Alzheimer's disease (Non-patent document 2) and Parkinson's disease (Non-patent document 3). However, it is unclear whether all non-steroidal anti-inflammatory drugs are effective for Alzheimer's disease and Parkinson's disease, and by what mechanism some non-steroidal anti-inflammatory drugs are effective against these diseases It is. Moreover, since it has been reported that cell death occurs when NSAIDs are used for gastric cells, overcoming such undesirable side effects is also a problem when using NSAIDs.

  On the other hand, the accumulation of unfolded proteins and aggregates in the ER results in ER stress. Moderate ER stress results in many rescue responses such as UPR (unfolded protein response) and degradation associated with ER (Non-Patent Document 4). UPR is characterized by activation of a signaling pathway triggered by IRE1, a kinase present in the ER (Non-Patent Document 5). Activation of IRE1 induces activation of X box binding protein (XBP-1) mRNA splicing. The spliced XBP-1 protein functions as a transcription factor and induces an ER stress gene (for example, GRP78 which is an ER molecular chaperone related to protein folding) (Non-patent Document 6). On the other hand, ER stress can induce cell death by increasing induction of CHOP, which is a proapoptotic transcription factor (Non-patent Documents 7 and 8).

However, it is not known whether non-steroidal anti-inflammatory drugs can alleviate ER stress so that ER-related diseases can be treated, and what non-steroidal anti-inflammatory drugs are ER stressed It is not even known whether it has an effect on.
CM Ulrich, J. Bigler, JD Potter, Non-steroidal anti-inflammatory drugs for cancer prevention: promise, perils and pharmacogenetics. Nat. Rev. Cancer 6 (2006) 130-140. PP Zandi, JC Breitner, Do NSAIDs prevent Alzheimer's disease? And, if so, why? The epidemiological evidence. Neurobiol. Aging 22 (2001) 811-817. M. Asanuma, I. Miyazaki, N. Ogawa, Neuroprotective effects of nonsteroidal anti-inflammatory drugs on neurodegenerative diseases. Curr. Pharm. Des. 10 (2004) 695-700. K. Mori, Tripartite management of unfolded proteins in the endoplasmic reticulum.Cell 101 (2000) 451-454. XZ Wang, HP Harding, Y. Zhang, EM Jolicoeur, M. Kuroda, D. Ron, Cloning of mammalian Ire1 reveals diversity in the ER stress responses.EMBO J. 17 (1998) 5708-5717. H. Yoshida, T. Matsui, A. Yamamoto, T. Okada, K. Mori, XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor.Cell 107 (2001) 881- 891. H. Zinszner, M. Kuroda, X. Wang, N. Batchvarova, RT Lightfoot, H. Remotti, JL Stevens, D. Ron, CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev. 12 (1998) 982-995. K. Hyoda, T. Hosoi, N. Horie, Y. Okuma, K. Ozawa, Y. Nomura, PI3K-Akt inactivation induced CHOP expression in endoplasmic reticulum-stressed cells. Biochem. Biophys. Res. Commun. 340 (2006) 286-290.

  It is an object of the present invention to provide a therapeutic agent for endoplasmic reticulum (ER) related diseases (for example, Alzheimer's disease, Parkinson's disease, and central nervous system (CNS) diseases such as cerebral ischemia).

  The above problem is that non-steroidal anti-inflammatory compounds such as pranoprofen are excellent therapeutic agents for ER-related diseases, such as Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes, cystic fibrosis. It was found and useful for the treatment of diseases of the central nervous system (CNS) (eg, nerve cells and glial cells) such as ischemic diseases and neurodegenerative diseases.

  Specifically, the present invention provides the following.

  (Item 1) A pharmaceutical composition for treating and / or preventing an endoplasmic reticulum stress-related disease in glial cells, comprising an effective amount of a non-steroidal anti-inflammatory compound, wherein the non-steroidal Anti-inflammatory compounds are pranoprofen, thiaprofenic acid, oxaprozin, loxoprofen sodium, aluminoprofen, zaltoprofen, diclofenac sodium, ampenac sodium, nabumetone, flurbiprofen, (s)-(+)-ibuprofen, naproxen, Acemetacin, indomethacin, indomethacin farnesyl, progouracin maleate, sulindac, etodolac, mofezolac, ampiroxicam, tenoxicam, meloxicam, lornoxicam, mefenamic acid, flufenamic acid, epili Lumpur, tiaramide hydrochloride, and is selected from the group consisting of emorfazone, pharmaceutical compositions.

  (Item 2) The endoplasmic reticulum stress-related disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes mellitus, cystic fibrosis, ischemic disease, neurodegenerative disease, obesity, manic depression, and cancer. The pharmaceutical composition according to claim 1.

  (Item 3) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is Alzheimer's disease.

  (Item 4) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is Parkinson's disease.

  (Item 5) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cerebral ischemia.

  (Item 6) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is diabetes.

  (Item 7) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cystic fibrosis.

  (Item 8) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is an ischemic disease.

  (Item 9) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is a neurodegenerative disease.

  (Item 10) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is obesity.

  (Item 11) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is manic depression.

  (Item 12) The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cancer.

(Item 13) Use of an effective amount of a non-steroidal anti-inflammatory compound for the manufacture of a therapeutic and / or prophylactic agent for endoplasmic reticulum stress-related diseases in nerve cells, wherein the non-steroidal anti-inflammatory is The active compound is pranoprofen, thiaprofenic acid, oxaprozin, loxoprofen sodium, aluminoprofen, zaltoprofen, diclofenac sodium, ampenac sodium, nabumetone, flurbiprofen, (s)-(+)-ibuprofen, naproxen, acemetacin, Indomethacin, indomethacin farnesyl, progourmetacin maleate, sulindac, etodolac, mofezolac, ampiroxicam, tenoxicam, meloxicam, lornoxicam, mefenamic acid, flufenamic acid, epilysole, Use, selected from the group consisting of tiaramid hydrochloride and emorphazone.

  (Item 14) The endoplasmic reticulum stress-related disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes mellitus, cystic fibrosis, ischemic disease, neurodegenerative disease, obesity, manic depression, and cancer. 14. Use according to claim 13, wherein:

  (Item 15) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is Alzheimer's disease.

  (Item 16) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is Parkinson's disease.

  (Item 17) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cerebral ischemia.

  (Item 18) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is diabetes.

  (Item 19) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cystic fibrosis.

  (Item 20) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is an ischemic disease.

  (Item 21) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is a neurodegenerative disease.

  (Item 22) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is obesity.

  (Item 23) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is manic depression.

  (Item 24) The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cancer.

  In accordance with the present invention, there are provided therapeutic agents for endoplasmic reticulum stress-related diseases and methods of using compounds for the manufacture of endoplasmic reticulum stress-related diseases.

  The present invention will be described below. Throughout this specification, it should be understood that the singular forms also include the plural concept unless specifically stated otherwise. In addition, it is to be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified. Thus, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

(Definition of terms)
Listed below are definitions of terms particularly used in the present specification.

  The term “planoprofen” as used herein refers to the following formula:

And the enantiomers and salts thereof.

  The term “diclofenac sodium” as used herein refers to the following formula:

And a salt thereof.

  The term “flurbiprofen” as used herein refers to the following formula:

And the enantiomers and salts thereof.

  As used herein, the term “acemetacin” refers to the following formula:

And a salt thereof.

  The term “mefenamic acid” as used herein refers to the following formula:

And a salt thereof.

  The term “epilysole” as used herein refers to the following formula:

And a salt thereof.

  The term “(s)-(+)-ibuprofen” as used herein refers to the following formula:

And the enantiomers and salts thereof.

  The term “thiaprofenic acid” as used herein refers to the following formula:

And a salt thereof.

  The term “oxaprozin” as used herein refers to the following formula:

And a salt thereof.

  The term “loxoprofen sodium” as used herein refers to the following formula:

And a salt thereof.

  The term “aluminoprofen” as used herein refers to the following formula:

And a salt thereof.

  The term “zaltoprofen” as used herein refers to the following formula:

And a salt thereof.

  As used herein, the term “ampenac sodium” refers to the following formula:

And a salt thereof.

  The term “nabumetone” as used herein refers to the following formula:

And a salt thereof.

  The term “naproxen” as used herein refers to the following formula:

And the enantiomers and salts thereof.

  As used herein, the term “indomethacin” refers to the following formula:

And a salt thereof.

  The term “indomethacin farnesyl” as used herein refers to the following formula:

And a salt thereof.

  As used herein, the term “progouramicin maleate” refers to the following formula:

And the enantiomers and salts thereof.

  The term “sulindac” as used herein refers to the following formula:

And a salt thereof.

  The term “etodolac” as used herein refers to the following formula:

And a salt thereof.

  The term “mofezolac” as used herein refers to the following formula:

And a salt thereof.

  The term “ampiroxicam” as used herein refers to the following formula:

And a salt thereof.

  The term “tenoxicam” as used herein refers to the following formula:

And a salt thereof.

  The term “meloxicam” as used herein refers to the following formula:

And a salt thereof.

  The term “lornoxicam” as used herein refers to the following formula:

And a salt thereof.

  The term “flufenamic acid” or “aluminum flufenamic acid” as used herein refers to the following formula:

And a salt thereof.

  As used herein, the term “tiaramid hydrochloride” refers to the following formula:

And a salt thereof.

  The term “emorphazone” as used herein refers to the following formula:

And a salt thereof.

  As used herein, the term “endoplasmic reticulum” refers to the reticulated spread of tubules or flat sac (tank) in a eukaryotic cell trade. Some endoplasmic reticulums have ribosomes attached and others do not.

  As used herein, the term “endoplasmic reticulum stress-related disease” refers to a disease caused by an endoplasmic reticulum abnormality, particularly an abnormality in protein biosynthesis. Examples of endoplasmic reticulum stress-related diseases include, but are not limited to, Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes, cystic fibrosis, ischemic disease, and neurodegenerative disease.

As used herein, the term “non-steroidal anti-inflammatory compound” refers to an anti-inflammatory agent having a cyclooxygenase inhibitory action, and the active ingredient of the anti-inflammatory agent having a cyclooxygenase inhibitory action is preferably a cyclooxygenase inhibitor. A compound having an IC ₅₀ value in a test of 100 μM or less (preferably 10 μM or less).

  As used herein, a “kit” includes a plurality of containers and manufacturer's instructions, and each container includes a pharmaceutical composition of the present invention, other agents, and a carrier. A product.

  As used herein, “pharmaceutically acceptable carrier” refers to a substance that is used when producing agricultural chemicals such as pharmaceuticals or animal drugs, and does not adversely affect active ingredients. Such pharmaceutically acceptable carriers include, for example, but are not limited to: antioxidants, preservatives, colorants, flavors, and diluents, emulsifiers, suspending agents, solvents , Fillers, bulking agents, buffering agents, delivery vehicles, excipients and / or pharmaceutical adjuvants.

  The type and amount of the drug used in the treatment method of the present invention is based on the information obtained by the method of the present invention (for example, information on the disease), the purpose of use, the target disease (type, severity, etc.), A person skilled in the art can easily determine the patient's age, weight, sex, medical history, the form or type of the site of the subject to be administered, and the like. The frequency with which the monitoring method of the present invention is applied to a subject (or patient) also depends on the purpose of use, target disease (type, severity, etc.), patient age, weight, gender, medical history, treatment course, etc. In view of this, it can be easily determined by those skilled in the art. The frequency of monitoring the disease state includes, for example, daily-once every several months (for example, once a week-once a month). It is preferable to perform monitoring once a week to once a month while observing the progress.

  If desired, more than one drug may be used in the treatment of the present invention. When two or more drugs are used, substances with similar properties or origins may be used, or drugs with different properties or origins may be used. Information regarding disease levels for methods of administering two or more such drugs can also be obtained by the methods of the present invention.

(Pharmaceutical composition)
In the present specification, the “effective amount” of a drug refers to an amount that allows the drug to exhibit the intended drug effect. In the present specification, among such effective amounts, the minimum concentration may be referred to as the minimum effective amount. Such minimum effective amounts are well known in the art, and usually the minimum effective amount of a drug is determined by those skilled in the art or can be determined as appropriate by those skilled in the art. In order to determine such an effective amount, an animal model or the like can be used in addition to actual administration. The present invention is also useful in determining such effective amounts.

The effective amount of pranoprofen in an adult (body weight 60 kg) is preferably 0.025 to 752 mg / kg / day, more preferably 0.25 to 75 mg / kg / day, even more preferably 0.83 to 22.5 mg / kg / day. The effective amount of thiaprofenic acid in adults (body weight 60 kg) is preferably 0.067-2004 mg / kg / day, more preferably 0.67-200 mg / kg / day, even more preferably 2.22-60 mg / day. kg / day. The effective amount of oxaprozin in an adult (body weight 60 kg) is preferably 0.13 to 2004 mg / kg / day, more preferably 1.33 to 200 mg / kg / day, even more preferably 4.44 to 60 mg / kg. / Day. The effective amount of loxoprofen sodium in an adult (body weight 60 kg) is preferably 0.02-601 mg / kg / day, more preferably 0.2-60 mg / kg / day, even more preferably 0.67-18 mg / day. kg / day. The effective amount of aluminoprofen in adults (body weight 60 kg) is preferably 0.067-2004 mg / kg / day, more preferably 0.67-200 mg / kg / day, even more preferably 2.22-60 mg / Kg / day. The effective amount of zaltoprofen in adults (body weight 60 kg) is preferably 0.026 to 802 mg / kg / day, more preferably 0.26 to 80 mg / kg / day, even more preferably 0.89 to 24 mg / kg. / Day. The effective amount of diclofenac sodium in an adult (body weight 60 kg) is preferably 0.008 to 334 mg / kg / day, more preferably 0.08 to 33 mg / kg / day, even more preferably 0.28 to 10 mg / kg / day. The effective amount of ampenac sodium in an adult (body weight 60 kg) is preferably 0.017 to 668 mg / kg / day, more preferably 0.17 to 67 mg / kg / day, even more preferably 0.56 to 20 mg. / Kg / day. The effective amount of nabumetone in adults (body weight 60 kg) is preferably 0.13 to 2672 mg / kg / day, more preferably 1.33 to 267 mg / kg / day, even more preferably 4.44 to 80 mg / kg. / Day. The effective amount of flurbiprofen in an adult (body weight 60 kg) is preferably 0.013-401 mg / kg / day, more preferably 0.13-40 mg / kg / day, even more preferably 0.44 ~ 12 mg / kg / day. The effective amount of ibuprofen in an adult (body weight 60 kg) is preferably 0.067-2004 mg / kg / day, more preferably 0.67-200 mg / kg / day, even more preferably 2.22-60 mg / day. kg / day. The effective amount of naproxen in adults (body weight 60 kg) is 0.13-2672 mg / kg / day, more preferably 1.33-267 mg / kg / day, even more preferably 4.44-80 mg / kg / day. It is.
The effective amount of acemetacin in adults (body weight 60 kg) is preferably 0.020-601 mg / kg / day, more preferably 0.20-60 mg / kg / day, even more preferably 0.67-18 mg / day. kg / day. The effective amount of indomethacin in adults (body weight 60 kg) is preferably 0.0083 to 251 mg / kg / day, more preferably 0.083 to 25 mg / kg / day, even more preferably 0.28 to 7.5 mg. / Kg / day. The effective amount of indomethacin farnesyl in adults (body weight 60 kg) is preferably 0.033 to 668 mg / kg / day, more preferably 0.33 to 67 mg / kg / day, even more preferably 1.11 to 20 mg / day. kg / day. The effective amount of progouritacin maleate in adults (body weight 60 kg) is preferably 0.03-902 mg / kg / day, more preferably 0.3-90 mg / kg / day, even more preferably 1-27 mg. / Kg / day. The effective amount of sulindac in an adult (body weight 60 kg) is preferably 0.017 to 668 mg / kg / day, more preferably 0.17 to 67 mg / kg / day, even more preferably 0.56 to 20 mg / kg. / Day. The effective amount of etodolac in an adult (body weight 60 kg) is preferably 0.033 to 1336 mg / kg / day, more preferably 0.33 to 133 mg / kg / day, even more preferably 1.11 to 7.5 mg. / Kg / day. The effective amount of mofezolac in an adult (body weight 60 kg) is preferably 0.025 to 751 mg / kg / day, more preferably 0.25 to 75 mg / kg / day, even more preferably 0.83 to 22.5 mg / Kg / day. The effective amount of ampiroxicam in adults (body weight 60 kg) is preferably 0.0045-90 mg / kg / day, more preferably 0.045-9 mg / kg / day, and even more preferably 0.15-2. 7 mg / kg / day. The effective amount of tenoxicam in an adult (body weight 60 kg) is preferably 0.0033 to 67 mg / kg / day, more preferably 0.033 to 6.67 mg / kg / day, even more preferably 0.11 to 2 mg. / Kg / day. The effective amount of meloxicam in an adult (body weight 60 kg) is preferably 0.0017-50 mg / kg / day, more preferably 0.017-5 mg / kg / day, even more preferably 0.056-1.5 mg. / Kg / day. The effective amount of lornoxicam in adults (body weight 60 kg) is preferably 0.0027-80 mg / kg / day, more preferably 0.027-8 mg / kg / day, even more preferably 0.089-2.4 mg. / Kg / day. The effective amount of mefenamic acid in adults (body weight 60 kg) is preferably 0.167-5010 mg / kg / day, more preferably 1.67-500 mg / kg / day, even more preferably 5.56-150 mg. / Kg / day. The effective amount of flufenamic acid in an adult (body weight 60 kg) is preferably 0.125 to 2505 mg / kg / day, more preferably 1.25 to 250 mg / kg / day, even more preferably 4.17 to 75 mg / day. kg / day. The effective amount of epirizole in an adult (body weight 60 kg) is preferably 0.050 to 1503 mg / kg / day, more preferably 0.50 to 150 mg / kg / day, even more preferably 1.67 to 45 mg / day. kg / day. The effective amount of tiaramid hydrochloride in adults (body weight 60 kg) is preferably 0.033-1002 mg / kg / day, more preferably 0.33-100 mg / kg / day, even more preferably 1.11-30 mg / day. kg / day. The effective amount of emorphazone in adults (body weight 60 kg) is preferably 0.067-2004 mg / kg / day, more preferably 0.67-200 mg / kg / day, even more preferably 2.22-60 mg / kg. / Day. As used herein, “pharmaceutically acceptable carrier” refers to a substance that is used when producing agricultural chemicals such as pharmaceuticals or animal drugs, and does not adversely affect active ingredients. Such pharmaceutically acceptable carriers include, for example, but are not limited to: antioxidants, preservatives, colorants, flavors, and diluents, emulsifiers, suspending agents, solvents , Fillers, bulking agents, buffering agents, delivery vehicles, excipients and / or agricultural or pharmaceutical adjuvants.

  The type and amount of the drug used in the treatment method of the present invention is based on the information obtained by the method of the present invention (for example, information on the disease), the purpose of use, the target disease (type, severity, etc.), A person skilled in the art can easily determine the patient's age, weight, sex, medical history, the form or type of the site of the subject to be administered, and the like. The frequency with which the monitoring method of the present invention is applied to a subject (or patient) also depends on the purpose of use, target disease (type, severity, etc.), patient age, weight, gender, medical history, treatment course, etc. In view of this, it can be easily determined by those skilled in the art. Examples of the frequency of monitoring the disease state include monitoring every day to once every several months (for example, once a week to once a month). It is preferable to perform monitoring once a week to once a month while observing the progress.

  If desired, more than one drug may be used in the treatment of the present invention. When two or more drugs are used, substances with similar properties or origins may be used, or drugs with different properties or origins may be used. Information regarding disease levels for methods of administering two or more such drugs can also be obtained by the methods of the present invention.

  In the present invention, once an analysis result of a specific sugar chain structure and a disease level are correlated with organisms, cultured cells, tissues, etc. of similar types (for example, mice against humans), the corresponding sugar Those skilled in the art will readily understand that the analysis results of the chain structure can be correlated with the disease level. Such matters are described and supported by, for example, the Animal Culture Cell Manual, edited by Seno et al., Kyoritsu Shuppan, 1993, etc., all of which are incorporated herein by reference.

(Prescription)
The invention also provides a method of treating and / or preventing a disease or disorder (eg, an infection) by administering an effective amount of a therapeutic agent to a subject. By therapeutic agent is meant a composition of the invention in combination with a pharmaceutically acceptable carrier type (eg, a sterile carrier).

  The therapeutic agent takes into account the clinical condition of the individual patient (especially the side effects of therapeutic agent alone treatment), the site of delivery, the method of administration, the dosage regimen and other factors known to those skilled in the art, eg, “Therapeutics Manual 2006”. Supervised by Fumiaki Takahisa, Yoshio Yazaki, edited by Akira Seki, Mitsuo Kitahara, Fumiaki Ueno, Hirotoshi Echizen. Therefore, the target “effective amount” in the present specification is determined by taking such consideration into consideration.

  As a general suggestion, the total pharmaceutically effective amount of the therapeutic agent administered parenterally per dose is in the range of about 1 mg / kg / day to 15 mg / kg / day of patient body weight, as described above. This is left to therapeutic discretion. More preferably, for the cell bioactive substances of the present invention, this dose is at least 1 mg / kg / day, most preferably between about 3 mg / kg / day and about 10 mg / kg / day for humans. Intravenous bag solutions may also be used. The duration of treatment necessary to observe the change and the post-treatment interval at which a response occurs appears to vary depending on the desired effect.

  Therapeutic agents are orally, rectally, parenterally, in the bath, vaginally, intraperitoneally, topically (such as by powders, ointments, gels, drops, or transdermal patches), by mouth, orally or by nasal spray Can be administered. “Pharmaceutically acceptable carrier” refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulant or any type of formulation adjuvant. The term “parenteral” as used herein refers to modes of administration including intravenous and intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injections and infusions.

  The therapeutic agents of the present invention are also suitably administered by a sustained release system. Suitable examples of sustained release therapeutic agents are oral, rectal, parenteral, intracisternal, intravaginal, intraperitoneal, topical (such as by powder, ointment, gel, infusion, or transdermal patch), It can be administered by mouth or as an oral or nasal spray. “Pharmaceutically acceptable carrier” refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulant or any type of formulation adjuvant. The term “parenteral” as used herein refers to modes of administration including intravenous and intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injections and infusions.

  The therapeutic agents of the present invention are also suitably administered by a sustained release system. Suitable examples of sustained release therapeutic agents include suitable polymer materials (eg, semipermeable polymer matrices in the form of molded articles (eg, films or microcapsules)), suitable hydrophobic materials (eg, in acceptable quality oils). Or an ion exchange resin, and poorly soluble derivatives (eg, poorly soluble salts).

  Sustained release matrices include polylactide (US Pat. No. 3,773,919, EP 58,481), a copolymer of L-glutamic acid and γ-ethyl-L-glutamate (Sidman et al., Biopolymers 22: 547-556 (1983)). ), Poly (2-hydroxyethyl methacrylate) (Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981), and Langer, Chem. Tech. 12: 98-105 (1982)), ethylene vinyl Acetate (Langer et al., Ibid.) Or poly-D-(-)-3-hydroxybutyric acid (EP133,988).

  Sustained release therapeutic agents also include the therapeutic agents of the present invention entrapped in liposomes (generally, Langer, Science 249: 1527-1533 (1990); Treat et al., Liposomes in the Disease of Infectious Disease and Cancer, Loop, -See Berstein and Fiddler (eds.), Liss, New York, pages 317-327 and 353-365 (1989)). Liposomes containing therapeutic agents can be prepared by methods known per se: DE 3,218,121; Epstein et al., Proc. Natl. Acad. Sci. USA 82: 3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77: 4030-4034 (1980); EP52,322; EP36,676; EP88,046; EP143,949; EP142,641; Japanese Patent Application No. 83-118008; US Pat. No. 4,485,045 And 4,544,545 and EP 102,324. Usually, liposomes are small (about 200-800 cm) unilamellar type, where the lipid content is greater than about 30 mol% cholesterol, and the selected proportion is adjusted for optimal therapeutic agents.

  In still further embodiments, the therapeutic agents of the invention are delivered by a pump (Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14: 201 (1987); Buchwald et al., Surgary 88: 507 ( 1980); Saudek et al., N. Engl. J. Med. 321: 574 (1989)).

  Other controlled release systems are discussed in the review by Langer (Science 249: 1527-1533 (1990)).

  For parenteral administration, in one embodiment, in general, the therapeutic agent is toxic to the recipient in the desired degree of purity, in a pharmaceutically acceptable carrier, i.e. the dosage and concentration used. It is formulated by mixing in a unit dosage injectable form (solution, suspension or emulsion) with one that is not and compatible with the other ingredients of the formulation. For example, the formulation preferably does not include oxidation and other compounds known to be harmful to therapeutic agents.

  Generally, formulations are prepared by contacting the therapeutic agent uniformly and intimately with liquid carriers or finely divided solid carriers or both. Next, if necessary, the product is shaped into the desired formulation. Preferably, the carrier is a parenteral carrier, more preferably a solution that is isotonic with the blood of the recipient. Examples of such carrier vehicles include water, saline, Ringer's solution, and dextrose solution. Nonaqueous vehicles such as fixed oils and ethyl oleate are also useful herein, as are liposomes.

  The carrier suitably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability. Such substances are not toxic to the recipient at the dosages and concentrations used, such as phosphate, citrate, succinate, acetic acid and other organic acids or their salts. Buffering agents; antioxidants such as ascorbic acid; low molecular weight (less than about 10 residues) polypeptides (eg, polyarginine or tripeptides); proteins such as serum albumin, gelatin or immunoglobulins; polyvinylpyrrolidone Hydrophilic polymers such as: amino acids such as glycine, glutamic acid, aspartic acid or arginine; monosaccharides, disaccharides and other carbohydrates including cellulose or derivatives thereof, glucose, mannose or dextrin; chelating agents such as EDTA Sugar sugars such as mannitol or sorbitol Lumpur; counterions such as sodium; and / or polysorbate include nonionic surfactants such as poloxamers or PEG.

  Any drug to be used for therapeutic administration can be free of organisms or viruses, ie, sterile. Aseptic conditions are easily achieved by filtration through a sterile filtration membrane (eg, 0.2 micron membrane). In general, the therapeutic agent is placed in a container having a sterile access port, for example, an intravenous solution bag or vial with a stopper puncturable with a hypodermic needle.

  The therapeutic agents are typically stored in unit dose or multi-dose containers, such as sealed ampoules or vials, as an aqueous solution or lyophilized formulation for reconstitution. As an example of a lyophilized formulation, a 10 ml vial is filled with 5 ml of a sterile filtered 1% (w / v) aqueous therapeutic agent and the resulting mixture is lyophilized. The lyophilized therapeutic agent is reconstituted with bacteriostatic water for injection to prepare an infusion solution.

  The present invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more components of the therapeutic agent of the present invention. A notice of the form prescribed by a government agency that regulates the manufacture, use or sale of a medicinal product or biological product may be attached to such a container, and this notice may be attached to the government regarding the manufacture, use or sale for human administration. Represents institutional approval. In addition, the therapeutic agent may be used in combination with other therapeutic compounds.

  The therapeutic agents of the present invention can be administered alone or in combination with other therapeutic agents. The combinations can be administered, for example, either simultaneously as a mixture; simultaneously or concurrently but separately; or over time. This includes the presentation that the combined agents are administered together as a therapeutic mixture, and also the procedure where the combined agents are administered separately but simultaneously, eg, through separate intravenous lines to the same individual . Administration "in combination" further includes separate administration of one of the compounds or agents given first, followed by the second.

  In certain embodiments, the pharmaceutical compositions of the invention are administered in combination with an anticancer agent.

  The non-steroidal anti-inflammatory drugs of the present invention can be administered alone or in combination with other therapeutic agents. The therapeutic agents that can be administered in combination with the therapeutic agents of the present invention include other anti-cancer agents, chemotherapeutic agents, antibiotics, steroidal anti-inflammatory agents, conventional immunotherapeutic agents, other cytokinins, and / or growth factors. For example, but not limited to. The combination can be administered, for example, either as a mixture simultaneously (simultaneously or concurrently but separately) or over time. This includes the presentation that the combined agents are administered together as a therapeutic mixture, and also the procedure where the combined agents are administered separately but simultaneously, eg, through separate intravenous lines to the same individual. Administration in combination further includes separate administration of one of the compounds given first, followed by the second, or drug.

  In a further embodiment, the non-steroidal anti-inflammatory drug of the invention is administered in combination with an antibiotic. Antibiotics that can be used include aminoglycoside antibiotics, polyene antibiotics, penicillin antibiotics, cephem antibiotics, peptide antibiotics, macrolide antibiotics, tetracycline antibiotics. It is not limited.

  In further embodiments, the therapeutic agents of the invention are administered in combination with other therapeutic or prophylactic regimes (eg, radiation therapy).

  Hereinafter, the present invention will be described in detail with reference to examples and the like, but the present invention is not limited thereto.

  The present invention will be described below. Throughout this specification, it should be understood that the singular forms also include the plural concept unless specifically stated otherwise. In addition, it is to be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified. Thus, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

(Materials and methods)
(Preparation of primary cultured glial cells)
Glial cells were prepared from whole brains of newborn C57BL / 6 mice. Specifically, the brain was removed from a newborn mouse (within 24 hours after birth) and the meninges were removed. The brain was permeated through a mesh (300 μm) and then incubated with DMEM containing 0.25% trypsin, 2000 Kunitz units of Deoxyribonuclease 1 to separate the cells. The cells were collected by centrifugation at 4 ° C. and 880 × g for 10 minutes, and were primarily cultured for 10 days at 37 ° C. and 95% air / 5% CO _{2 in} DMEM medium containing 10% FCS. Thereafter, the mixed glial cells were shaken at 120 rpm for 18 hours, suspended in PBS (-) containing 1 mM EGTA, and further cultured for 5 to 7 days, and used as mixed glial cells in the experiment. These cells contain 95% or more of GFAP-positive astrocytes and about 5% of microglia positive for ED1 (anti-macrophage / microglia mAb) staining.

(RT-PCR analysis)
Total RNA was isolated using TRI reagent (Sigma-Aldrich, St. Loius, MO) and RT-PCR was performed. Specifically, 25 U Superscript Reverse transcriptase 3 (Invitrogen) and Oligo (Tvitrogen) and Oligo (Tvitrogen) ₁₂ in a 20 μl reaction mixture containing Superscript buffer (Invitrogen), 1 mM dNTP mix, 10 mM DTT, and 20 U RNase inhibitor. CDNA was synthesized from total RNA by reverse transcription using ₁₈ primers (Invitrogen). Total RNA and oligo (dT) _12-18 primers were incubated at 70 ° C. for 10 minutes prior to the reverse transcription reaction. After 1.5 hours incubation at 46 ° C., the reaction was terminated by denaturing the enzyme for 15 minutes at 70 ° C. For PCR amplification, 1.2 μl of cDNA was added to 12 μl of reaction mixture containing 0.2 μM of each primer, 0.2 μd dNTP mix, 0.6 U Taq polymerase, and reaction buffer. PCR was performed in a DNA thermal cycler (GeneAmp® PCR system 9700). The following primers were used: XBP-1 upstream (5′-cct tgt ggt tga gaa cca gg-3 ′; SEQ ID NO: 1); XBP-1 downstream (5′-cta gag gct tgg tgt data c-3 ′ SEQ ID NO: 2); GRP78 upstream (5′-ctg ggt aca ttt gat ctg act gg-3 ′; SEQ ID NO: 3); GRP78 downstream (5′-gca tcc tgg tgg ctt tcc agg cat tc-3 ′; SEQ ID NO: 4); CHOP upstream (5′-ccc tgc ctt tca cct tgg-3 ′; SEQ ID NO: 5); CHOP downstream (5′-ccg ctc gtt ctc ctg ctc-3 ′; SEQ ID NO: 6); GAPDH upstream (5 ′ -Aaa ccc atc acc atc ttt cag-3 '; SEQ ID NO: 7); GAPDH downstream (5'-agg gc cat cca cag tct tct-3 '; SEQ ID NO: 8). PCR products (10 μl) were run on an 8% polyacrylamide gel in TBE buffer. The gel was stained with ethidium bromide and photographed under ultraviolet irradiation. XBP-1, GRP78, CHOP, and GAPDH cDNA were circulated in 35 cycles (94 ° C. 1 minute, 55 ° C. 1 minute, 72 ° C. 1 minute), 18 cycles (94 ° C. 1 minute, 55 ° C. 1 minute, 72 ° C. 1 respectively). Min), 22 cycles (94 ° C 1 minute, 57 ° C 1 minute, 72 ° C 1 minute) and 18 cycles (94 ° C 1 minute, 57 ° C 1 minute, 72 ° C 1 minute), and these PCR reaction products Were electrophoresed separately. These cycle numbers were determined based on preliminary experiments that determined the linearity range of the amplification reaction for each molecule.

(Western blot reaction)
Western blot was performed as follows. The cells were washed with ice-cold PBS and 10 mM HEPES-NaOH (pH 7.5), 150 mM, 1 mM EGTA, 1 mM Na ₃ VO ₄ , 10 mM NaF, 10 μg / ml aprotinin, 10 μg / ml leupeptin, 1 mM PMSF, And dissolved in a buffer containing 1% NP-40 for 20 minutes. The lysate was centrifuged at 15,000 rpm for 20 minutes at 4 ° C. and the supernatant was collected. Samples were boiled for 3 minutes with Ramli buffer, electrophoresed on SDS-PAGE and transferred to a nitrocellulose membrane at 4 ° C. Membranes were incubated with anti-KDEL antibody (StressGen; 1: 1,000 dilution), anti-CHOP antibody (Santa Cruz, 1: 500 dilution), and then with anti-horseradish peroxidase-conjugated antibody. Peroxidase was detected by chemiluminescence using the ECL system (Amersham).

(Statistical analysis)
Results were expressed as mean ± SE. Statistical analysis was performed using a paired t test.

Example 1: Planoprofen inhibits GRP78 and CHOP expression induced by ER stress
Experiments were conducted on the effect of pranoprofen on GRP78 and CHOP expression induced by ER stress. In this example, tunicamycin, an inhibitor of protein N-glycosylation, was used to induce ER stress. Cells were pretreated with pranoprofen (1 mM) for 1 hour and then stimulated with tunicamycin (10 ng / ml). As assessed by RT-PCR, we observed an increase in GRP78 transcripts that were significantly inhibited by pranoprofen (FIGS. 1A and B). As assessed by Western blot, the expression of GRP78 significantly inhibited by pranoprofen was increased by tunicamycin (FIGS. 1C and D). On the other hand, pranoprofen alone did not increase GRP78 expression (FIG. 1). Similarly, CHOP expression that was significantly inhibited by pranoprofen at both mRNA and protein levels was induced by ER stress (FIG. 2). Planoprofen alone did not affect CHOP expression in primary glial cells (FIG. 2). These results indicate that pranoprofen has an inhibitory effect on GRP78 and CHOP expression induced by ER stress.

Example 2: Effect of pranoprofen on eIF2α activation induced by ER stress
CHOP expression induced by ER stress has been reported to mediate through a pathway mediated by PERK-eIF2α (Harding et al., Mol. Cell 6 (2000) 1099-1108; and Schuneer et al., Mol. Cell 7 (2001) 1165-1176). Therefore, it was next determined whether pranoprofen activates eIF2α. Treatment with tunicamycin (10 ng / ml) for 2 hours increased eIF2α phosphorylation (FIG. 3). Interestingly, as shown in FIG. 3, pranoprofen treatment alone increased eIF2α phosphorylation and the extent of phosphorylation was further increased by ER stress. Therefore, these results indicated that the effect of pranoprofen on CHOP expression induced by ER stress is an event independent of the activation state of eIF2α.

Example 3 Effect of Planoprofen on XBP1 Splicing Induced by ER Stress
ER stress has been reported to induce XBP1 splicing. The spliced truncated form of XBP1 functions as a transcription factor involved in GRP78 or CHOP induction. Therefore, it was determined whether or not pranoprofen affects XBP1 splicing induced by ER stress. Treated with pranoprofen for 1 hour, then stimulated with tunicamycin (10 ng / ml) for 6 hours, and RT-PCR analysis was performed. Planoprofen alone did not induce XBP1 splicing (FIG. 4). However, as shown in FIG. 4, pranoprofen significantly inhibited XBP1 splicing induced by ER stress. These results indicated that pranoprofen specifically inhibited XBP1 splicing induced by ER stress.

(Discussion)
Recent evidence suggests that NSAIDs have a novel pharmacological action in addition to the cyclooxygenase inhibitory action. In the present invention, a novel pharmacological action of pranoprofen was found that affects the ER stress response in glial cells. Epidemiological studies have shown that the use of NSAIDs reduces neurodegenerative diseases such as Alzheimer's disease. However, the mechanism of NSAIDs for neurodegenerative diseases is unclear. The results of the present invention show one possible mechanism for NSAIDs that can affect ER stress.

  The unique properties of pranoprofen against ER stress were observed. Planoprofen inhibited splicing of XBP1, induced by ER stress, GRP78 expression and CHOP expression, but increased eIF2α phosphorylation. The results of the present invention differ from previous observations that reported that NSAIDs induced genes (eg, GRP78 and CHOP) induced by ER stress, resulting in gastric cell death. On the other hand, NSAIDs have been reported to inhibit cell death in SH-SY5Y neuronal cells induced by ER stress. In the present invention, the inhibitory effect of planoprofen on GRP78 expression and CHOP expression in glial cells induced by ER stress was observed. The reason for the discrepancy between the results of the present invention and past reports is unclear, but is thought to be due to the difference in the origin of the cells used in the experiment and / or the type of NSAIDs used. Supporting this conclusion is the fact that the effects of NSAIDs are not necessarily the same for each individual NSAIDs and show different pharmacological effects depending on the dose used and the target tissue. . The gastrointestinal side effects of NSAIDs may explain the apoptotic effect on gastric cells. On the other hand, the observed effect of pranoprofen on the ER stress of SH-SY5Y, which is a neuronal cell, or the inhibitory effect of NSAIDs on the ER stress response of glial cells observed in the present invention is related to the administration of NSAIDs. It can explain the reduced risk of degenerative diseases.

  From these results, pranoprofen is a therapeutic agent for endoplasmic reticulum stress-related diseases in nerve cells (eg, Alzheimer's disease, Parkinson's disease, and cerebral ischemia) without causing harmful side effects such as cell death. It is concluded that it is useful.

(Example 3: Treatment and / or prevention of endoplasmic reticulum stress-related diseases with nonsteroidal anti-inflammatory compounds other than pranoprofen)
Treatment of endoplasmic reticulum stress-related diseases for the non-steroidal anti-inflammatory compounds other than pranoprofen, diclofenac sodium, flurbiprofen, acemetacin, mefenamic acid, epilysole, and (s)-(+)-ibuprofen Tested for prevention.

  Primary glial cells were treated with various non-steroidal anti-inflammatory compounds (DSS: diclofenac sodium, FLU: flurbiprofen, ACE: acemetacin, MEF: mefenamic acid, MEP: epirizole, and IBU: (s)-(+)- Pretreated with ibuprofen, 100 μM each). In the sample marked “+ Tm”, stimulation with tunicamycin (Tm, 10 ng / ml) for 6 hours after pretreatment with non-steroidal anti-inflammatory compounds. “Tm” is the result of not using a non-steroidal anti-inflammatory compound, and “Cont” is the result of using neither a non-steroidal anti-inflammatory compound nor tunicamycin. In “Cont”, DMSO, which is a solvent of Tm, is treated.

  The results of Western analysis using GRP78 specific antibody and CHOP specific antibody are shown. FIG. 5 is a result showing the protein amount of GRP78 as a result of measuring the concentration of the sample on the gel (relative value assuming 1 for GRP78). FIG. 6 is a result showing the protein amount of CHOP as a result of measuring the concentration of the sample on the gel (relative value with 1 for CHOP).

  As shown in FIG. 5 and FIG. 6, any of the non-steroidal anti-inflammatory compounds used suppressed the endoplasmic reticulum stress response. Therefore, it can be concluded that these non-steroidal anti-inflammatory compounds are also useful as therapeutic agents for endoplasmic reticulum stress-related diseases (for example, Alzheimer's disease, Parkinson's disease, and cerebral ischemia), like pranoprofen.

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, this invention should not be limited and limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. It is understood that those skilled in the art can implement an equivalent range based on the description of the present invention and the common general technical knowledge from the description of specific preferred embodiments of the present invention. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

  According to the present invention, a novel therapeutic agent for endoplasmic reticulum stress-related diseases (eg, Alzheimer's disease, Parkinson's disease, and cerebral ischemia) in nerve cells is provided.

FIG. 1 shows the results of pranoprofen suppressing GRP78 expression induced by ER stress. FIG. 1A shows the results of pretreatment of glial cells with pranoprofen (Pra, 1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 6 hours. The results of RT-PCR analysis using GRP78-specific primers and GAPDH-specific primers are shown. FIG. 1B is a result showing the amount of GRP78 mRNA as a result of measuring the concentration of a sample on a gel (relative value with GAPDH as 1). FIG. 1C shows the results of pretreatment of glial cells with pranoprofen (1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 18 hours. The levels of GRP78 and GAPDH were determined by Western blot. FIG. 1D is a result showing the level of GRP78 protein as a result of measuring the concentration of a sample on a gel (relative value with GAPDH as 1). Values were expressed as mean ± standard deviation (n = 3 per group). ^* P <0.05 (significant difference between Tm and Tm + planoprofen). FIG. 2 shows the result of pranoprofen suppressing CHOP expression induced by ER stress. FIG. 2A shows the results of pretreatment of glial cells with pranoprofen (1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 6 hours. The results of RT-PCR analysis performed using CHOP specific primers and GAPDH specific primers are shown. FIG. 2B is a result showing the amount of CHOP mRNA as a result of measuring the concentration of a sample on a gel (relative value with GAPDH as 1). FIG. 2C shows the results of pretreatment of glial cells with pranoprofen (1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 18 hours. The levels of CHOP and GAPDH were determined by Western blot. FIG. 2D is a result showing the CHOP protein level as a result of measuring the concentration of the sample on the gel (relative value with GAPDH as 1). Values were expressed as mean ± standard deviation (n = 3 per group). ^* P <0.05 (significant difference between Tm and Tm + planoprofen). FIG. 3 shows the results of pranoprofen increasing eIF2α phosphorylation induced by ER stress. FIG. 3A shows the results of pretreatment of glial cells with pranoprofen (1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 2 hours. The level of phosphorylated eIF2α (Ser51) and the level of GAPDH were determined by Western blot. FIG. 3B shows the result of showing the level of phosphorylated eIF2α (Ser51) as a result of measuring the concentration of the sample on the gel (relative value assuming 1 for GAPDH). Values were expressed as mean ± standard deviation (n = 3 per group). ^* P <0.05 (significant difference between Tm and Tm + planoprofen). FIG. 4 shows the results of pranoprofen suppressing XBP1 splicing induced by ER stress. FIG. 4A shows the results of pretreatment of glial cells with pranoprofen (1 mM) for 1 hour and then stimulation with tunicamycin (Tm, 10 ng / ml) for 6 hours. The results of RT-PCR analysis using XBP1-specific primers and GAPDH-specific primers are shown. “U-XBP1” indicates an unspliced transcript, and “s-XBP1” indicates a spliced transcript. FIG. 4B is a result showing the amount of XBP1 mRNA as a result of measuring the concentration of a sample on a gel (relative value with GAPDH as 1). Values were expressed as mean ± standard deviation (n = 3 per group). ^* P <0.05 (significant difference between Tm and Tm + planoprofen). FIG. 5 shows GRP78 expression induced by ER stress with non-steroidal anti-inflammatory compounds other than pranoprofen (diclofenac sodium, flurbiprofen, acemetacin, mefenamic acid, epilysole, and (s)-(+)- The result which ibuprofen) suppresses is shown. The results are shown in which primary glial cells were pretreated with each non-steroidal anti-inflammatory compound (100 μM) for 1 hour and then stimulated with tunicamycin (Tm, 10 ng / ml). The result of having performed western analysis using the GRP78 specific antibody is shown. The figure shows the result of showing the protein mass of GRP78 as a result of measuring the concentration of the sample on the gel (relative value assuming 1 for GRP78). Cont: no non-steroidal anti-inflammatory compound or tunicamycin added, DSS: diclofenac sodium, FLU: flurbiprofen, ACE: acemetacin, MEF: mefenamic acid, MEP: epirizole, IBU: (s)-(+)- Ibuprofen, Tm: tunicamycin, DSS + Tm: diclofenac sodium and tunicamycin, FLU + Tm: flurbiprofen and tunicamycin, ACE + Tm: acemetacin and tunicamycin, MEF + Tm: mefenamic acid and tunicamycin, MEP + Tm: epirisol and tunicamycin, and I +)-Ibuprofen and tunicamycin. FIG. 6 shows the expression of CHOP induced by ER stress in non-steroidal anti-inflammatory compounds other than pranoprofen (DSS: diclofenac sodium, FLU: flurbiprofen, ACE: acemetacin, MEF: mefenamic acid, MEP: epirizole , And IBU: (s)-(+)-ibuprofen) show the results of inhibition. The results are shown in which primary glial cells were pretreated with each non-steroidal anti-inflammatory compound (100 μM) for 1 hour and then stimulated with tunicamycin (Tm, 10 ng / ml). The results of Western analysis using a CHOP specific antibody are shown. The figure shows the results of the protein amount of CHOP as a result of measuring the concentration of the sample on the gel (relative value assuming 1 for CHOP). Cont: no non-steroidal anti-inflammatory compound or tunicamycin added, DSS: diclofenac sodium, FLU: flurbiprofen, ACE: acemetacin, MEF: mefenamic acid, MEP: epirizole, IBU: (s)-(+)- Ibuprofen, Tm: tunicamycin, DSS + Tm: diclofenac sodium and tunicamycin, FLU + Tm: flurbiprofen and tunicamycin, ACE + Tm: acemetacin and tunicamycin, MEF + Tm: mefenamic acid and tunicamycin, MEP + Tm: epirisol and tunicamycin, and I +)-Ibuprofen and tunicamycin.

  SEQ ID NO: 1 is the sequence of the XBP-1 upstream primer.

  SEQ ID NO: 2 is the sequence of the XBP-1 downstream primer.

  SEQ ID NO: 3 is the sequence of the GRP78 upstream primer.

  SEQ ID NO: 4 is the sequence of the GRP78 downstream primer.

  SEQ ID NO: 5 is the sequence of the CHOP upstream primer.

  SEQ ID NO: 6 is the sequence of the CHOP downstream primer.

  SEQ ID NO: 7 is the sequence of the GAPDH upstream primer.

  SEQ ID NO: 8 is the sequence of the GAPDH downstream primer.

Claims (24)

A pharmaceutical composition for the treatment and / or prevention of endoplasmic reticulum stress-related disease in glial cells, comprising an effective amount of a non-steroidal anti-inflammatory compound, wherein said non-steroidal anti-inflammatory compound Are pranoprofen, thiaprofenic acid, oxaprozin, loxoprofen sodium, aluminoprofen, zaltoprofen, diclofenac sodium, ampenac sodium, nabumetone, flurbiprofen, (s)-(+)-ibuprofen, naproxen, acemetacin, indomethacin, Indomethacin farnesyl, progourmet tacin maleate, sulindac, etodolac, mofezolac, ampiroxicam, tenoxicam, meloxicam, lornoxicam, mefenamic acid, flufenamic acid, epilysole, hydrochloric acid Aramid, and it is selected from the group consisting of emorfazone, pharmaceutical compositions. The endoplasmic reticulum stress-related disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes, cystic fibrosis, ischemic disease, neurodegenerative disease, obesity, manic depression, and cancer. Item 2. A pharmaceutical composition according to Item 1. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is Alzheimer's disease. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is Parkinson's disease. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cerebral ischemia. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is diabetes. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cystic fibrosis. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is an ischemic disease. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is a neurodegenerative disease. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is obesity. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is manic depression. The pharmaceutical composition according to claim 2, wherein the endoplasmic reticulum stress-related disease is cancer.
Use of an effective amount of a non-steroidal anti-inflammatory compound for the manufacture of a therapeutic and / or prophylactic agent for endoplasmic reticulum stress-related diseases in nerve cells, wherein the non-steroidal anti-inflammatory compound comprises: Planoprofen, thiaprofenic acid, oxaprozin, loxoprofen sodium, aluminoprofen, zaltoprofen, diclofenac sodium, ampenac sodium, nabumetone, flurbiprofen, (s)-(+)-ibuprofen, naproxen, acemetacin, indomethacin, indomethacin farnesyl , Progouritacin maleate, sulindac, etodolac, mofezolac, ampiroxicam, tenoxicam, meloxicam, lornoxicam, mefenamic acid, flufenamic acid, epilysole, tiaramid hydrochloride , And it is selected from the group consisting of emorfazone, used. The endoplasmic reticulum stress-related disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, cerebral ischemia, diabetes, cystic fibrosis, ischemic disease, neurodegenerative disease, obesity, manic depression, and cancer. Item 14. Use according to Item 13. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is Alzheimer's disease. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is Parkinson's disease. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cerebral ischemia. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is diabetes. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cystic fibrosis. The use according to claim 14, wherein the endoplasmic reticulum stress-related disease is an ischemic disease. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is a neurodegenerative disease. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is obesity. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is manic depression. 15. Use according to claim 14, wherein the endoplasmic reticulum stress-related disease is cancer.