CN1791394A - Preventive and/or therapeutic agent for inflammatory bowel disease - Google Patents

Preventive and/or therapeutic agent for inflammatory bowel disease Download PDF

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Publication number
CN1791394A
CN1791394A CNA2004800135747A CN200480013574A CN1791394A CN 1791394 A CN1791394 A CN 1791394A CN A2004800135747 A CNA2004800135747 A CN A2004800135747A CN 200480013574 A CN200480013574 A CN 200480013574A CN 1791394 A CN1791394 A CN 1791394A
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enteropathy
inflammatory bowel
gga
mice
heat shock
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浅香正博
西平顺
大川原辰也
武田宏司
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R&amp D Management Co., Ltd.
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Eisai Co Ltd
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    • 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
    • A61K31/12—Ketones
    • A61K31/121—Ketones acyclic
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Abstract

A medical agent that is available for bowel diseases, especially inflammatory bowel disease. In particular, a preventive and/or therapeutic agent for bowel diseases, comprising as an active ingredient a heat shock protein inducer or its salt, or a hydrate thereof. Specifically, there is provided a preventive and/or therapeutic agent for bowel diseases, comprising a prenyl ketone compound, 6,10,14,18-tetramethyl-5,9,13,17-nanodecatetraen-2-one, as an active ingredient.

Description

Inflammatory bowel prevents and/or treats agent
Technical field
The present invention relates to the agent that prevents and/or treats of inflammatory bowel, more specifically relate to the agent that prevents and/or treats of inflammatory bowel such as ulcerative colitis or clone disease.
Background technology
People's inflammatory bowel (Inflammatory Bowel Disease, abbreviate " IBD " below as) be the mucosa generation chronic inflammatory disease or the ulcer of large intestine and small intestinal, the diseases associated with inflammation that is chronic process, the cause of disease of this disease are not also known (for example, referring to non-patent literature 1).Above-mentioned IBD is except that comprising ulcerative colitis or clone disease, broadly also comprise intestinal type behcets disease or ischemic enteritis etc., above-mentioned ulcerative colitis is mainly attacked big intestinal mucosa, be a kind of non-special inflammation of agnogenic diffusivity that forms erosion or ulcer, above-mentioned clone disease is a diseases associated with inflammation of following fibrosis or ulcer and nonspecific granuloma.At present, in Japan, the patient of above-mentioned disease is more and more, according to healing property of the MHLW's refractory intestinal tube barrier study class report the year before last, ulcerative colitis, clone disease have involved totally 10 ten thousand people, particularly in more than 10 year old～more than 20 year old youngster, see more, can't effect a radical cure at present, all will struggle against, be a kind of difficult disease of misery lifetime with above-mentioned disease.
The mechanism of action of inflammation and the immunne response of IBD are also indeterminate, but clear and definite at present tumor necrosis factor (below be called " TNF ") α or macrophage migration stop various inflammatory mediators such as the factor with pathological changes or worsen relevant.(for example, referring to non-patent literature 2 to 5).
At present mainly be to use 5-aminosalicylic acid (5-aminosalicylicacid, abbreviate as " 5-ASA "), glucocorticoid, immunosuppressant or immunoregulatory agent at the treatment of IBD.But, also have the patient who above-mentioned treatment is had patience.Recently, as new therapeutic agent, developed anti-TNF-alpha monoclonal antibodies (for example, referring to non-patent literature 6 and 7).Above-mentioned antibody suppresses the activity of TNF-α, improves clone disease patient's enteritis symptom fast.But, usually side effect such as accompanying infection and malignant tumor (for example, referring to non-patent literature 8 and 9).Above-mentioned side effect makes the treatment of anti-TNF-alpha monoclonal antibodies be difficult to continue.
Heat shock protein (below abbreviate " HSP " as) is the stress-induced type protein (for example, referring to non-patent literature 10) that shows protective nature, control immunne response.The major function of HSP is as playing a role at chaperone (chaperone) in the cell of folding unusually or variant protein matter, under stressed condition, gives cell protection function.A kind of as multiple HSP has because of molecular weight to be about the HSP70 that 70kDa gains the name.In its subfamily, have at the protein that acts on the very strong cytoprotective function of stress having of stomach, liver and heart (for example, referring to non-patent literature 11 to 14).The HSP70 transgenic mice is to heart and injury models show opposing type (for example, referring to non-patent literature 15 and 16).In people's intestinal, ulcerative colitis compares with the non-specific colitis of big intestinal mucosa, and the expression of HSP70 increases (referring to non-patent literature 17).But the effect in the progression of disease of IBD is not also studied fully for HSP.
But geranyl geranyl acetone (geranyl geranyl acetone) (below abbreviate " GGA " as) is the non-ring type polyisoamylene chemical compound that the protection stomach is avoided the ulcer equivalent damage.Known this chemical compound does not influence gastric acid secretion, protect effectively gastric mucosa avoid various stress (for example, referring to non-patent literature 18 to 20).In addition, the not only synthetic or secretion gastric mucin (referring to non-patent literature 21) of GGA also strengthens the synthetic or secretion (referring to non-patent literature 22) of glycoprotein or surface activity phospholipid composition.Several years ago, reported the expression (referring to non-patent literature 23) that GGA induces HSP70 in the gastric mucosal cell.And, the GGA oral administration, the many places local organization is induced the expression of HSP70 in vivo, and protective tissue avoids destroying and inflammation (referring to non-patent literature 24 to 26).
In addition, submitted to record GGA to the effective patent application of inflammatory bowel (referring to patent documentation 1), but just in the claim scope, put down in writing, in detailed explanation, do not put down in writing fully and show that the geranyl geranyl acetone is to the effective experimental example of inflammatory bowel, so whether effectively also indeterminate to inflammatory bowel.
In order to study the factor of inducing enteral inflammation or immunne response, a lot of experimental animal models have been utilized.In above-mentioned model, consider the reliability and the convenience of model itself, extensively utilize dextran sulfate sodium (below abbreviate " DSS " as) or 2,4,6-trinitro-benzene-sulfonic acid (below abbreviate " TNBS " as) is induced enteritis model (for example, referring to non-patent literature 27 to 29).
On the other hand; as same gastric mucosa protectant; known Ecabet Sodium (ecabetsodium); though known its as prevention, the therapeutic agent (for example) of inflammatory bowel of the present invention referring to patent documentation 2, the chemical constitution of Ecabet Sodium and GGA and dissmilarity; and because oral administration is difficult to arrive lower digestive tract such as large intestine; so, shown in several experimental examples in the above-mentioned patent documentation 1, need the coloclysis administration at least in order to be used for the treatment of inflammatory bowel.Under the situation of described coloclysis, because directly administration around the inflammation part, therefore except that bring very big burden to the patient, also exist because of medicine to arrive the problem that a part of falling large intestine or horizontal large intestine limits therapeutic domain.
Non-patent literature 1:Podolsky DK.Inflammatory bowel disease.N Engl JMed 2002; 347:417-29.
Non-patent literature 2:Sartor RB.Pathogenesis and immune mechanisms ofchronic inflammatory bowel disease.Am J Gastroenterol 1997; 92 (12Suppl.): 5S-11S.
Non-patent literature 3:Papadakis KA, Targan SR.Role of cytokines in thepathogenesis of inflammatory bowel disease.Annu Rev Med 2000; 51:289-98.
Non-patent literature 4:de Jong YP, Abadia-Molina AC, Satoskar AR, et al.Development of chronic colitis is dependent on the cytokine MIF.2001; 2:1061-66.
Non-patent literature 5:Ohkawara T, Nishihira J, Takeda H, et al.Ameliorationof dextran sulfate sodium-induced colitis by anti-macrophage migrationinhibitory factor antibody in mice.Gastroenterology 2002; 123:256-70.
Non-patent literature 6:van Dullemen HM, van Deventer SJ, Hommes DW, et al.Treatment of Crohn ' s disease with anti-tumor necrosis factor chimericmonoclonal antibody (cA2) .Gastroenterology 1995; 109:129-35.
Non-patent literature 7:Targan SR, Hanauer SB, van Deventer SJ, et al.Ashort-term study of chimeric monoclonal antibody cA2to tumor necrosisfactor alpha for Crohn ' s disease.Crohn ' s Disease cA2Study Group.N Eng JMed 1997; 337:1029-35.
Non-patent literature 8:Bickston SJ, Lichtenstein GR, Arseneau KO, et al.Therelationship between infliximab treatment and lymphoma in Crohn ' s disease.Gastroenterology 1999; 117:1433-37.
Non-patent literature 9:Keane J, Gershon S, Wise RP, et al.Tuberculosisassociated with infliximab, a tumor necrosis factor alpha-neutralizing agent.N Engl J Med 2001; 345:1098-104.
Non-patent literature 10:Pockley AG.Heat shock protein as regulators of theimmune response.Lancet 2003; 9; 362 (9382): 469-76.
Non-patent literature 11:Nakamura K, Rokutan K, Marui N, et al.Induction ofheat shock proteins and their implication in protection againstethanol-induced damage in cultured guinea pig gastric mucosal cells.Gastroenterology 1991; 101:161-166.
Non-patent literature 12:Mestril R, Chi SH, Sayen MR, et al.Expression ofinducible stress protein 70 in rat heart myogenic cells confers protectionagainst stimulated ishemia-induced injury.J Clin Invest 1994; 93:759-767.
Non-patent literature 13:Urayama S, Musch MW, Retsky J, et al.Dexamethasone protection of rat IEC against oxidant injury mediated byinduction ofheat shock protein 72.J Clin Invest 1998; 102:1860-1865.
Non-patent literature 14:Fujimori S, Otaka M, Otani S, et al.Induction of a72-kDa heat shock protein and cytoprotection against thioacetamide-inducedliver injury in rats.Dig Dis Sci 1997; 42:1987-1994.
Non-patent literature 15:Marber MS, Mestril R, Chi SH, et al.Overexpression ofthe rat inducible 70-kD heat stress protein in a transgenic mouse increasesthe resistance of the heart to ischemic injury.J Clin Invest 1995; 95:1446-56.
Non-patent literature 16:Hutter JJ, Mestril R, Tam EK, et al.Overexpression ofheat shock protein 72 in transgenic mice decreases infarct size in vivo.Circulation 1996; 94:1408-11.
Non-patent literature 17:Ludwig D, Stahl M, Ibrahim ET, et al.Enhancedintestinal expression of heat shock protein 70 in patients with inflammatorybowel disease.Dig Dis Sci1999; 44:1440-47.
Non-patent literature 18:Terano A, Shiga J, Hiraishi H, et al.Protective action oftetraprenylacetone against ethanol-induced damage in rat gastric mucosa.Digestion 1986; 35:182-88.
Non-patent literature 19:Pappas TN, Mulvihill SJ, Goto Y, et al.Advances indrug therapy for peptic ulcer disease.Arch Surg 1987; 122:447-50.
Non-patent literature 20:BilskiJ, Sarosiek J, Murty VL, et al.Enhancement ofthe lipid content and physical properties of gastric mucus bygeranylgeranylacetone.BiochemPharmacol 1987; 36:4059-65.
Non-patent literature 21:Fujimoto M, Yamanaka T, Bessho M, et al.Effects ofgeranylgeranylacetone on gastrointestinal secresion.Eur J Pharmacol 1982; 77:113-18.
Non-patent literature 22:Ito M, Tanaka T, Suzuki Y.Increasing action ofteprenone, a new antiulcer agent, on high-molecular-weight glycoprotein ingastric mucus during the healing process of acetic acid-induced ulcer in rats.Jpn J Pharmocol 1986; 41:117-25.
Non-patent literature 23:Hirakawa T, Rokutan K, Nikawa T, et al.Geranylgeranylacetone induces heat shock proteins in cultured guinea piggastric mucosal cells and rat gastric mucosa.Gastroenterology 1996; 111:345-57.
Non-patent literature 24:Ooie T, Takahashi N, Saikawa T, et al.Single oral doseof Geranylgeranylacetone induces heat-shock protein 72 and rendersprotection against ischemia/reperfusion injury in rat heart.Circulation 2001; 104:1837-43.
Non-patent literature 25:Yamagami K, Yamamoto Y, Ishikawa Y, et al.Effectsof geranyl-geranyl-acetone administration before heat shock preconditioningfor conferring tolerance against ischemia-reperfusion injury in rat livers.JLab Clin Med 2000; 135:465-75.
Non-patent literature 26:Unoshima M, Iwasaka H, Eto J, et al.Antiviral effectsof geranylgeranylacetone:enhancement of MxA expression andphosphorylation of PKR during influenza virus infection.Antimicrob AgentsChemother 2003; 47:2914-21.
Non-patent literature 27:Elson CO, Sartor RB, Tennyson GS, et al.Experimentalmodels of inflammatory bowel disease.Gastroenterology 1995; 109:1344-67.
Non-patent literature 28:Okayasu I, Hatakeyama S, Yamada M, et al.A novelmethod in the induction of reliable experimental acute and chronic ulcerativecolitis in mice.Gastroenterology 1990; 98:694-702.
Non-patent literature 29:Morris GP, Beck PL, Herridge MS, et al.Hapten-induced model of chronic inflammation and ulceration in the ratcolon.Gastroenterology 1989; 96:795-803.
Patent documentation 1:WO02/098398 communique
Patent documentation 2: the spy opens the 2002-104962 communique
Summary of the invention
The purpose of this invention is to provide a kind of new formulation, said preparation does not bring very big burden and safe and effective in the preventing and/or treating of the inflammatory bowel of whole intestinal to the patient.
The inventor etc. further investigate, and found that specific non-annularity cluster isoprenoid induces HSP in intestinal, are effectively for preventing and/or treating of inflammatory bowel, thereby have realized the present invention.
That is, first kind of scheme of the present invention provides the hydrate that contains heat shock protein derivant or its salt or above-claimed cpd to prevent and/or treat agent as the enteropathy of effective ingredient.
The preferred version that enteropathy of the present invention prevents and/or treats agent is that above-mentioned heat shock protein derivant is isoamyl ketenes (prenylketone) compounds.
The preferred version that enteropathy of the present invention prevents and/or treats agent is that above-mentioned iso-amylene ketone compounds is 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
Figure A20048001357400111
The preferred version that enteropathy of the present invention prevents and/or treats agent is that above-mentioned enteropathy is an inflammatory bowel.
The preferred version that enteropathy of the present invention prevents and/or treats agent is that above-mentioned inflammatory bowel is the disease that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
The preferred version that enteropathy of the present invention prevents and/or treats agent is that above-mentioned heat shock protein is a heat shock protein 70.
In addition, the invention provides with 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as the preventive and/or the therapeutic agent of the inflammatory bowel of effective ingredient.
Figure A20048001357400112
The inflammatory bowel of preventive of the present invention and/or therapeutic agent is the disease that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
In addition, second kind of scheme of the present invention provides the enteropathy of the step that comprises the hydrate that gives heat shock protein derivant or its salt or above-claimed cpd to prevent and/or treat method.
The preferred version that enteropathy of the present invention prevents and/or treats method is that above-mentioned heat shock protein derivant is the iso-amylene ketone compounds.
The preferred version that enteropathy of the present invention prevents and/or treats method is that above-mentioned iso-amylene ketone compounds is 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
The preferred version that enteropathy of the present invention prevents and/or treats method is that above-mentioned enteropathy is an inflammatory bowel.
The preferred version that enteropathy of the present invention prevents and/or treats method is that above-mentioned inflammatory bowel is the disease that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
The preferred version that enteropathy of the present invention prevents and/or treats method is that above-mentioned heat shock protein is a heat shock protein 70.
In addition, the invention provides with 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as the prevention method and/or the Therapeutic Method of the inflammatory bowel of effective ingredient.
Prevention method of the present invention and/or the inflammatory bowel that Therapeutic Method was suitable for are the diseases that comprises clone disease, ulcerative colitis, intestinal behcets disease and ischemic enteritis.
And, the hydrate that the third scheme of the present invention provides heat shock protein derivant or its salt or above-claimed cpd prevents and/or treats purposes in the agent in the preparation enteropathy, and above-mentioned enteropathy prevents and/or treats agent and contains the hydrate of heat shock protein derivant or its salt or above-claimed cpd as effective ingredient.
The preferred version of such use of the present invention is that above-mentioned heat shock protein derivant is the iso-amylene ketone compounds.
The preferred version of such use of the present invention is that above-mentioned iso-amylene ketone compounds is 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
The preferred version of such use of the present invention is that above-mentioned enteropathy is an inflammatory bowel.
The preferred version of such use of the present invention is that above-mentioned inflammatory bowel is the disease that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
The preferred version of such use of the present invention is that above-mentioned heat shock protein is a heat shock protein 70.
In addition, the hydrate that the invention provides heat shock protein derivant or its salt or above-claimed cpd is in the preventive of preparation inflammatory bowel and/or the purposes in the therapeutic agent, the preventive of above-mentioned inflammatory bowel and/or therapeutic agent contain 6 shown in the formula (I), 10,14,18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as effective ingredient.
Figure A20048001357400132
Diseases associated with inflammation in the purposes of the present invention comprises the disease of clone disease, ulcerative colitis, intestinal behcets disease and ischemic enteritis.
According to the present invention, chemical compound 6,10,14 of the present invention, 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is induced heat shock protein 70 in intestinal, for enteropathy, be effective for preventing and/or treating of inflammatory bowel more specifically.In addition, because chemical compound of the present invention can not pass through the coloclysis administration, and carry out oral administration, so can under the situation of not bringing undue burden, prevent and/or treat enteropathy to the patient.
Description of drawings
Fig. 1 represents that the GGA oral administration is to the result of the clinical effectiveness of DSS administration after 7 days among the present invention.The data of diarrhoea and hemorrhage of rectum are by the F check analysis.The delta data of body weight loss utilizes the Studentt check analysis.All values all compare with the mice that DSS and contrast medicine are handled.In addition, *Expression P＜0.05, *Expression P＜0.01, * *Expression P＜0.001, a represents with administration to be the same day 100% o'clock ratio.
Fig. 2 represents the result of geranyl geranyl acetone (GGA) to the inhibition effect that contains large intestine short and smallization of 3% dextran sulfate sodium (DSS) administration after 7 days in the drinking water.The rod figure on the left side represents the result of normal control mice, and the result of the mice of contrast medicine is given in intermediary excellent figure expression, and the rod figure on the right represents the result with the mice of the GGA processing of 300mg/kg.Give to contrast the mice (n=10) of medicine and the difference of the mice (n=10) that GGA handles statistical significance (p＜0.05) is arranged.Same result obtains confirming in 3 independent experiments.
Fig. 3 represents that GGA induces the histology of the effect of BALB/c mouse tissue injury that the result is discussed to DSS.Big intestinal tissue is launched along long axis direction, fix, be immersed in the paraffin then with 10% neutral buffered formalin.After from microscope slide, removing deparaffnize the very thin tissue slice, sample is dyeed with HE.(A) the mice large intestine microphotograph of drinking water is only given in expression.(B) expression 7 days mice large intestine microphotograph of 3%DSS solution-treated.Observing struvite cellular infiltration of serious symptom and epithelial cell destroys.(C) expression contrasted the mice large intestine microphotograph of medicine to the mice that gives 3%DSS solution every 1 day.Present the histological appearance identical with (B).(D) the mice large intestine microphotograph the mice that gives 3%DSS solution handled with the GGA of 300mg/kg every (totally 4 times) on the 1st of expression.GGA suppresses the struvite cellular infiltration in mucosa and the submucous tissue, and demonstrates the big intestinal epithelial cell of protection and avoid destructive effect.In addition, the multiplying power among this figure is 100 times.
Fig. 4 represents the large intestine histological score result of the mice of 3%DSS administration after 7 days.In the Histological evaluation of tissue injury,, utilize the method described in the embodiment of this description quantitative with microscopic evaluation struvite lesion degree.Rod figure with point represents the result to the mice (n=10) of contrast medicine, the result of the mice (n=10) that black rod figure expression GGA handles.The result of the result of the mice (n=10) that GGA handles and the mice (n=10) of giving the contrast medicine relatively, tissue injury and lesion degree are all had a mind to the free burial ground for the destitute and are suppressed.
Fig. 5 is illustrated in the large intestine that 3%DSS induces colitis, HSP25,40,70,90 Western Blot analysis result.The large intestine tissue homogenate is by preparing with the large intestine of the mice of administration after 12 hours before the GGA administration.Utilize the SDS-PAGE isolated protein, be transferred to then on the NC Nitroncellulose, use HSP or beta-actin specific antibody immunoblotting HSP25,40,70,90 expression.Compare with the mice of giving the contrast medicine, the HSP70 of the mice that GGA handles expresses significantly to be increased.Same result independently obtains confirming in the experiment at 3 times.
Fig. 6 represents that DSS induces the immunohistochemical analysis result of HSP70 in the large intestine of colitis.Represent large intestine photo (50 times) with the mice of anti-HSP70 antibody or sheep IgG processing.(A) the big intestinal mucosa section photo of the normal mouse of handling with sheep IgG is taken from expression.(B) the large intestine section photo with the normal mouse of anti-HSP70 antibody treatment is taken from expression.(C) the large intestine section photo of mice is handled in expression with the vehicle (vehicle) of anti-HSP70 antibody treatment.(D) the large intestine section photo of the mice of handling with DSS and GGA is taken from expression.Compare with untreated fish group and contrast medicine administration group mice, the expression of HSP70 increases in superficial epithelium cell and the infiltration mononuclear cell.Microphotograph is represented the representative section that obtained by experiment.All test portions have same outward appearance.
Fig. 7 represents mice after 3%DSS induces colitis to handle 7 days, the active result of the myeloperoxidase (MPO) of large intestine (MPO).The rod figure on the left side represents the result of normal control mice, and intermediary rod figure represents the result of vehicle processing mice, and black rod figure expression is through the result of the mice of the GGA of 300mg/kg processing.The oral administration of GGA can suppress because of giving active raising of MPO that DSS causes.The difference that the mice (n=10) of contrast medicine administration group and GGA handle mice (n=10) has statistical significance.
Fig. 8 represents after 3%DSS handles 7 days, the TNF-α of large intestine and the generation result of IFN-γ.Utilize the elisa assay cytokine content described in the embodiment of this description.The rod figure on the left side represents the result of normal control mice, and intermediary rod figure represents the result of vehicle processing mice, and black rod figure expression is through the result of the mice of the GGA of 300mg/kg processing.TNF-α in the large intestine of the mice (n=10) that GGA handles and the generation of IFN-γ are compared with contrast medicine administration group mice (n=10), and reduce the free burial ground for the destitute intentionally.
Fig. 9 represents the influence result of GGA to the inductive serious symptom colitis of TNBS survival rate.GGA estimates after 7 days the influence of the survival rate of the BALB/c mouse handled with TNBS (270mg/kg).The rod figure on the left side represents the result of normal control mice, and intermediary rod figure represents the result through the mice of TNBS and vehicle processing, and black rod figure expression is through the result of the mice of the GGA of TNBS and 300mg/kg processing.The result who obtains by 3 independent experiments with on average ± SE represents.
Figure 10 represents that GGA induces the prevention result of the body weight loss of colitis to TNBS.Expression GGA induces the protection effect of the body weight loss that causes after 7 days to TNBS.The result of the undressed mice of white rod figure expression (n=10), the rod figure with oblique line represent the result of the mice (n=6) of handling through TNBS and vehicle, the result of the mice (n=6) that black rod figure expression is handled through the GGA of TNBS and 300mg/kg.The result with on average ± SE represents.
Figure 11 is the pathological tissue figure of the matched group of DSS enteritis model mice.
Figure 12 is the pathological tissue figure of the GGA administration group of DSS enteritis model mice.
The specific embodiment
Following embodiment is for example of the present invention is described, is not that the present invention is defined in this embodiment.The present invention only otherwise break away from its main idea can implement in every way.
The invention provides preventive effective to the treatment of enteropathy, especially inflammatory bowel, that toxicity is little and/or therapeutic agent.Describe the present invention below in detail.
Chemical compound as effective ingredient of the present invention is the iso-amylene ketone compounds of representing with formula (I).
The chemistry of the preferred chemical compound that uses is called 6,10,14 among the present invention, 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone (another name: the geranyl geranyl acetone, common name: teprenone abbreviates " GGA " below as).The GGA that the present invention uses has the two keys in 4 places in its structure, one co-exists in 8 kinds of geometric isomers, but is not particularly limited among the present invention, can use any isomer, also can be the mixture more than 2 kinds or 2 kinds.In the above-claimed cpd, as preferred chemical compound can enumerate (5E, 9E, 3E)-6,10,14,18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone and (5Z, 9E, 13E)-6,10,14,18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
The GGA that the present invention uses is known chemical compound, can be used as reagent, the raw material of industry is bought.As the synthetic method of GGA, for example, it is synthetic to open in the clear 53-145922 communique disclosed method according to the spy.
Can there be polymorphic among the effective ingredient GGA that uses among the present invention, be not particularly limited, can use any crystal type separately, also can use the crystal type mixture.And the metabolite that the GGA that uses among the present invention decomposes generation in vivo is also included within the claim scope of the present invention.
Preventive of the present invention and/or therapeutic agent can directly use GGA, or cooperate following usually as the composition of the raw material of pharmaceutical preparations, make preparation by common method, mentioned component comprises that known pharmacology goes up acceptable carrier etc., for example, excipient (specifically can be enumerated lactose, white sugar, starch, mannitol etc.), binding agent (specifically can be enumerated alphalise starch, arabic gum, carboxycellulose, polyvinyl pyrrolidone etc.), lubricant (specifically comprises magnesium stearate, Talcum etc.), disintegrating agent, coloring agent, drug flavoring, stabilizing agent, emulsifying agent, absorption enhancer, surfactant, pH adjusts agent, antiseptic, antioxidant etc.In addition, can also cooperate compositions such as blood flow ameliorant, antibacterial, antiinflammatory, cell activator, vitamins, aminoacid, wetting agent, keratin-lytic agent as required.
As the dosage form of the preparationization of preventive of the present invention and/or therapeutic agent, can enumerate tablet, powder, granula subtilis, granule, coated tablet, capsule, syrup, contain tablet, inhalant, suppository, injection, lyophilized preparation, ointment, Eye ointments, eye drop, nasal drop, ear drop, paste, lotion etc.
In addition, in the present invention, the administering mode of GGA is not particularly limited preferred oral administration.GGA can buy with the product of commodity " Selbex " (Eisai Co., Ltd's system) by name.
(for example, people, mice, rat, Cavia porcellus, rabbit, Canis familiaris L., horse, monkey etc.) preventing and/or treating of enteropathy is useful, and be particularly useful to treating and/or preventing of people's enteropathy to mammal for preventive that contains GGA of the present invention and/or therapeutic agent.
The preventive of the GGA of containing of the present invention and/or the dosage of therapeutic agent are suitably selected according to conditions such as symptom, age, body weight, the 20～2000mg on the one that is grown up, preferred 50～1000mg, further preferred 100～500mg.
The disease of GGA compounds for treating of the present invention is an enteropathy, can enumerate clone disease, ulcerative colitis, intestinal type behcets disease, simple ulcer, lonizing radiation enteritis and ischemic enteritis etc., and is especially effective to clone disease or ulcerative colitis.
Utilize the following examples to illustrate in greater detail the present invention, but scope of the present invention is not limited thereto.Based on explanation of the present invention, those skilled in the art can carry out various changes, modification, and above-mentioned change, modification are also included among the present invention.
Embodiment
Embodiment 1
(laboratory animal)
Mice is used SPF (specific pathogen free) the BALB/c male mice (Charles River Japan (strain)) of 8-10 age in week, body weight 22～25g.All mice all gives the experiment feedstuff of standard, makes its drinking-water of freely ingesting.The present invention observes Declaration of Helsinki, has obtained the approval that Hokkaido University's medical board zoopery ethics is stipulated committee.
(statistical analysis)
Unless otherwise specified, total data described later is all represented with meansigma methods ± standard deviation.(Cray NC), carries out statistical analysis to the result for StatView, SAS Institute to use F check and Student t check.P＜0.05 is thought statistical significance.
(preparation of GGA and oral administration)
Use after GGA (trade name " Selbex ", Eisai Co., Ltd's system) mixed by 5% gumwater and 0.0008% tocopherol.By the metal tube that has on the 1ml syringe,, will be expressed as 5ml/kg/ time GGA per os with volume and give mice with 300mg/kg/ time dosage.Control group mice gives to contain the arabic gum (hereinafter referred to as the contrast medicine) of the tocopherol of same amount.Each organizes mice after preceding 2 hours of DSS administration and DSS administration for the first time, continuous 7 days, every other day gives GGA or contrast medicine once (totally 4 times).In the clinical evaluation of DSS colitis, dose dependent experiment is that the different dosing amount (50,100,300,500mg/kg) by GGA is estimated.For utilizing Western Blot method to analyze the induce effect of GGA, mice is handled with the administration of 300mg/kg GGA single oral HSP70.Collect the sample of the big intestinal tissue of each mice of GGA or contrast medicine administration front and back.
(dextran sulfate sodium: DSS colitis inducing and estimating)
In 7 days, freely take in 3.0%DSS (molecular weight: 40,000 by making mice; ICNBiochemicals Inc system, California, USA) aqueous solution is induced enteritis.From administration, observe body weight, hemorrhage of rectum and the diarrhoea of mice every day.The body weight of each mice of weighing.In each stage of experiment, for carrying out Histological evaluation, measure the activity of cytokine, HSP and myeloperoxidase (MPO) (below be called " MPO "), with the big intestinal tissue of various interval collections.Large intestine is kept at-80 ℃ before using.The order of severity of colitis is estimated by large intestine length and histological examination after beginning 7 days in the DSS administration.
Fig. 1 represents in an embodiment of the present invention, and the mice of contrast medicine administration group and GGA processed group is suffered from diarrhoea when DSS induces colitis and the frequency of hemorrhage of rectum.As shown in Figure 1, with contrast medicine administration group relatively, suffer from diarrhoea in the GGA processed group of 300mg/kg/ time and 500mg/kg/ time and the frequency of hemorrhage of rectum is had a mind to the free burial ground for the destitute minimizing.In addition, in contrast medicine administration group, with the body weight before the DSS administration as 100%, after the DSS administration 7 days, mice loses weight to 86.3 ± 2.4%, and in the GGA processed group, compares with processed group, the variation of body weight is had a mind to the free burial ground for the destitute and is reduced (300mg/kg/ time group is 93.8 ± 4.1%, and 500mg/kg/ time group is 94.7 ± 5.0%).In the clinical evaluation of GGA dose dependent effect, find that GGA300mg/kg/ time dosage just can fully suppress seizure of disease.
There is close relation in short and smallization of known large intestine with the variation of tissue, usually the length of large intestine is used as the morphology parameter of inflammation degree.In order to estimate the length of large intestine, respectively organize mice in official hour execution.Fig. 2 represents the length of DSS administration large intestine after 7 days.Fig. 2 represents the large intestine length of GGA processed group, and (6.8 ± 0.3cm) have a mind to the free burial ground for the destitute is longer than the large intestine length (5.1 ± 0.1cm) of contrast medicine processed group.
(histologic analysis of DSS colitis)
Big intestinal tissue is cut and expansion along long axis direction, fix, be immersed in the paraffin with 10% neutral formalin buffer.Behind the paraffin of removing above-mentioned tissue part on the microscope slide, with hematoxylin eosin (hereinafter referred to as " HE ") dyeing.Histological evaluation as tissue injury, utilize the struvite lesion region of microscopic examination, method (Benjamin IJ, McMIllan DR.Stress (heat shock) proteins molecular chaperones.Circ Res.199827 to Cooper etc.; 83:117-32.) carry out some modifications, undertaken quantitatively by two pathology experts.Big damage of intestines is divided into 6 stages.That is, 0: mucosa is normal, and 1: inflammatory cell infiltration, 2: the following gland nest of half or half short and smallization takes place, 3: gland nests more than half short and smallization take place, 4: the gland nest disappears, and 5: epithelial cell shedding (ulcer or erosion).In addition, go back the scope of assess inflammation pathological changes.The extent of disease of large intestine integral body is divided into 6 stages.That is, 0:0%, 1:1～20%, 2:21～40%, 3:41～61%, 4:61～80%, 5:81～100%.
Fig. 3 represents the histologic analysis result of DSS colitis of the present invention.The large intestine histology of the BALB/c mouse that DSS handles utilizes HE dyeing, and GGA studies the effect of tissue injury in order to estimate.Large intestine soaks into the number minimum (referring to Fig. 3 A) before DSS handles of cell, and DSS handled after 7 days, and the number that soaks into cell increases, and the gland nest disappears simultaneously, downright bad increase (referring to Fig. 3 B) of epithelial cell.In the GGA processed group, tissue injury obtains free burial ground for the destitute improvement intentionally, compares with contrast medicine processed group, soaks into the decreased number (referring to Fig. 3 C and D) of inflammatory cell.In the mice that GGA handles, comprise that the histopathology appraisal result of tissue injury and extent of disease compares with untreated fish group or contrast medicine processed group mice, having a mind to the free burial ground for the destitute reduces (referring to Fig. 4).In addition, Fig. 4 represents the mice large intestine histological score result of 3%DSS administration of the present invention after 7 days.
(Western Blot analysis)
Use Polytron homogenizer (Kinematica, Lucerne Switzerland) to pulverize tissue.The tissue homogenate of equivalent is dissolved among 20 μ l 50mM (pH6.8) Tris-HCl that contain 2 mercapto ethanol (1%), SDS (2%), glycerol (20%) and bromophenol blue (0.04%), sample was heated 5 minutes at 100 ℃.Next, utilize SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to separate above-mentioned sample, then, be transferred on the NC Nitroncellulose film through electrophoresis.With the fixing above-mentioned film of 1% fat-free dry breast in the PBS solution, use anti-HSP25,40,70,90 antibody (Stressgen, Victoria, BC Canada) and beta-actin (Sigma, St Loius, MO) survey, make itself and the goat anti-rabbit igg antibody reaction that is combined with horseradish peroxidase (hereinafter referred to as " HRP ") then.In order to be used for checkout system, residual complex is handled based on manufacturer's operation indication.Use Micro BCA protein to check the protein concentration of the quantitative cell tissue homogenate of test kit.
(immunohistochemistry)
The immunohistochemical analysis of HSP70 is based on manufacturer's operation indication, uses the VectastainABC test kit to carry out.The big intestinal tissue of parafin bath is cut into the thick thin slice of 4 μ.With above-mentioned thin slice at 3%H 2O 2In, carry out pre-treatment in 10 minutes in 4 ℃, next, at room temperature, the normal sheep serum with 10% was handled 30 minutes, cultivated for 1 night at 4 ℃ with anti-HSP70 antibody (diluting Stressgen, Victoria, BC Canada with 1: 200) then.Utilize diaminobenzidine (diaminobenzidine) with the HSP70 positive staining, make it visual.
Fig. 5 is in the large intestine of the colitis model mice that 3%DSS handles, HSP25,40,70,90 Western Blot analysis result.Use the antibody of HSP25,40,70,90 correspondences, utilize the expression of Western Blot analysis and research HSP.HSP70 is a little less than GGA adds that (300mg/kg) is preceding and detects, but its expression increases after GGA handles 12 hours.On the other hand, HSP25,40 and 90 expression are handled through GGA and not to be changed (referring to Fig. 5).
Fig. 6 represents that DSS of the present invention induces the immunohistochemical analysis result of HSP70 in the large intestine of colitis.In order to confirm to suffer from the expressive site of HSP70 in the large intestine of mice that DSS induces colitis, utilize the specific antibody of HSP70 in the big intestinal tissue to carry out the immunohistology evaluation of HSP70.The immunoreactivity of HSP70 is the weak positive (referring to Fig. 6 B) at the gland nest place of epithelial cell and normal mouse.Induce and contrast medicine at DSS and handle that the male staining cell of HSP70 exists hardly in the large intestine of mice.What is interesting is that more the dyeing strong a lot (referring to Fig. 6 D) of mice is handled in the dyeing comparison of the big intestinal tissue of mice that GGA handles according to medicine.Above-mentioned phenomenon has been supported GGA induces endogenous HSP at large intestine hypothesis.
(mensuration of activity of myeloperoxidase in the large intestine)
Organizing the MPO activity is according to method (the Krawisz JE with Krawisz etc., Sharon P, Stenson WF.Quantitative assay for acute intestinal inflammation based onmyeloperoxidase activity.Assessment of inflammation in rat and hamstermodels.Gastroenterology 1984; 87:1344-50.) carry out that some amended methods measure.That is, use Polytron type homogenizer, each test portion is placed ice bath respectively, in buffer (in pH6.0, the 50mM kaliumphosphate buffer, 0.5% cetyl trimethyl ammonium bromide), pulverize and organize test portion (about 300mg), carry out each 30 seconds altogether 3 times.4 ℃ with 20, the condition of 000xg is above-mentioned sample centrifugalize 20 minutes, collects supernatant.The above-mentioned sample of 100 μ l is joined 2.9ml contain in the 50mM phosphate buffer (pH6.0) of the 3,3'-dimethoxy-4,4'-diaminobiphenyl. hydrochlorate of 0.167mg/ml and 0.0005% hydrogen peroxide, measure at 25 ℃ of use beam splitters.In order to proofread and correct, (Bio-rad laboratories, Hercules CA) determine the protein concentration of supernatant, and use and derive from human leukocyte (refining MPO MO) is worth standardization with it for Sigma, St.Loius to use the Bradford test kit.
As other parameters of inflammation, the activity level of big intestinal tissue the marrow being invaded by pathogen peroxidase (MPO) is arranged.MPO is the enzyme that is mainly produced by multinuclear leucocyte, is the relevant enzyme of granulocyte with tissue.Fig. 7 represents that 3%DSS induces colitis to handle the active result of MPO of the mice large intestine after 7 days among the present invention.Vehicle in the DSS administration is handled in the mice, and the MPO activity level enlarges markedly (referring to Fig. 7).(2.8 ± 0.3units/g) relatively, and the MPO activity level that GGA handles mice is had a mind to the free burial ground for the destitute and reduced (1.4 ± 0.4units/g, P＜0.03) with the MPO activity level of vehicle processed group.The above results has confirmed that GGA hinders granulocyte and soaks into.
(at the enzyme immunoassay of cytokine)
Utilize TNF-alpha specific ELISA test kit (GT, Minneapolis, MN) the TNF-α in the big intestinal tissue of mensuration.The anti-mice TNF-α IgG monoclonal antibody that is dissolved in the phosphate buffer normal saline (PBS) of 50ml is added in each hole of 96 orifice plates, placed 30 minutes in room temperature.Behind above-mentioned plate usefulness distilled water flushing 3 times,, placed 20 minutes in room temperature all adding the PBS that contains 0.5% bovine serum albumin (BSA) that is used for fixing in the holes.After removing fixed solution, add 2 blood plasma or tissue sample, in incubated at room temperature 1 hour to each hole.The syringe that heparin has been added in use utilizes cardiac puncture to gather blood plasma.Be preparation large intestine tissue sample, with Polytron homogenizer (Kinematica, Lucerne Switzerland) pulverizing contains protease inhibitor (according to manufacturer's operation indication modulation, tissue among the PBS of mixture every 20mg tissue use 1 μ l protease inhibitor), then with 12, the condition centrifugalize of 000xg 10 minutes detects supernatant.Utilize mice reorganization TNF-alpha protein in order to obtain standard curve.After 3 orifice plates of PBS flushing that contain 0.05%Tween-20 (dcq buffer liquid), in each hole, add the anti-TNF-Alpha antibodies that 50 μ l are combined with biotin., with dcq buffer liquid orifice plate is washed 3 times once more after 1 hour in incubated at room temperature.Next, in each hole, add the goat anti-rabbit igg antibody be combined with avidin, with above-mentioned orifice plate in incubated at room temperature 15 minutes.After dcq buffer liquid flushing 3 times, in each hole, add 50 μ l matrix solutions.Contain 8 μ g o-phenylenediamines and 4 μ l 30%H in the above-mentioned 10 μ l matrix solutions 2O 2Citric acid phosphoric acid buffer (pH5.0).In incubated at room temperature after 20 minutes, with the 4N sulphuric acid stopped reaction of 25 μ l.Use ELISA orifice plate reader (Bio-rad, Model 3550, Hercules, CA USA) measures absorbance under 492nm.
Equally, the mice IFN-γ in the tissue also uses IFN-γ specific ELISA test kit to measure.
Be to handle and suppress the viewpoint illustration position that cytokine generates the large intestine, use ELISA to measure the generation of TNF-α and IFN-γ in the big intestinal tissue from GGA.Fig. 8 represents that 3%DSS handles the generation result of TNF-α and IFN-γ in the large intestine after 7 days.In big intestinal tissue, the content of TNF-α and IFN-γ increases after DSS handles 7 days (be respectively 7.9 ± 1.0 and 9.7 ± 1.2pg/mg albumen).On the other hand, GGA generation and the vehicle of handling the above-mentioned cytokine of mice is handled mice and is compared intentionally the free burial ground for the destitute and reduce (be respectively 4.2 ± 0.8 and 4.1 ± 0.8pg/mg albumen).The above results hint GGA suppresses the generation of short scorching (proinflammatory) cytokine in the large intestine.
(2,4,6-trinitro-benzene-sulfonic acid: the inducing and estimating of TNBS colitis)
Make above-mentioned BALB/c mouse induce the TNBS colitis.That is, fasting pneumoretroperitoneum benzene injection at one night barbital (70mg/kg), anesthetized mice.Has polyethylene intubate (intramedic PE-20 for estimating the survival rate of TNBS colitis, utilizing; Beckton Dicknson society system; Jennings technology) 1ml syringe is to 100 μ l TNBS (250mg/kg) in intestinal tube inner chamber (apart from the about 3cm of anus outer rim) injection 50% alcoholic solution.For measuring the variation that body weight was compared with first day, be the TNBS processing mice of 100mg/kg with the dosage in 50% the ethanol.
For estimating the effect of GGA in the different colitis models, studied GGA TNBS has been induced the survival rate of colitis and the effect of body weight loss.In this model, because the TNBS dosage of 270mg/kg is induced the serious symptom inflammation, therefore a lot of mices are dead after 7 days in the TNBS administration.
Fig. 9 represents the influence result of GGA of the present invention to the survival rate of the inductive serious symptom colitis of TNBS.The contrast medicine is handled and the survival rate of TNBS processing mice is reduced to 23.3 ± 8.8% (referring to Fig. 9).And by the GGA oral administration, TNBS induces the survival rate of colitis to have a mind to the free burial ground for the destitute and increases to 56.7 ± 3.3%.
Figure 10 represents that GGA induces the result of preventive effect of the body weight loss of colitis to TNBS.Identical with survival rate, handle mice relatively (22.1 ± 3.3%) with the contrast medicine, GGA handles the body weight loss minimum (5.6 ± 1.2%) (referring to Figure 10) of mice.
Embodiment 2
(appearance of colitis in the DSS model)
Make the mice (Balb/c mice, Charles River, Shizuoka) in 8 ages in week (n=10) in 7 days, freely drink 3% dextran sulfate sodium (hereinafter referred to as " DSS ") aqueous solution, enteritis occurs.Mice occurs suffering from diarrhoea, having blood in stool, lose weight DSS aqueous solution administration the 5th～6 day.Before the DSS administration and DSS administration after 3 days, use sniffer that GGA (Eisai Co., Ltd's system) 100mg/kg is blended in administration in the normal saline.
(the GGA administration suppresses effect to the inflammation of colitis model)
The DSS administration begins to get the mice intestinal tube after 7 days, and the naked eyes evaluation of tissue is learned phenomenon.The pathological tissue figure of non-administration group and administration group is respectively as Figure 11 and shown in Figure 12.Consequently in the matched group of Figure 11 tissue injury taking place, on the other hand, does not almost see tissue injury in GGA administration group.And, at the 6 stages scoring that the inventor formulates, degree of tissue damage, inflammation scope that inflammation causes are marked respectively.The scoring benchmark is as follows.
The degree of tissue damage that inflammation causes
0: NIP
1: inflammatory cell slightly soaks into
2: slight short and smallization of gland nest
3: short and smallization of gland nest height
4: the gland nest disappears
5: epithelial cell shedding
The scope of inflammation
0: do not have
1：1～20％
2：21～40％
3：41～60％
4：61～80％
5：81～100％。
The scoring of GGA administration group and non-administration group is compared, and its result is as shown in table 1.As shown in Table 1, give the degree that GGA can suppress tissue damage, and dwindle the scope of inflammation.In addition, similarly compare diarrheal sickness rate, the sickness rate of having blood in stool, the rate of losing weight, intestinal tube length.Its result is as shown in table 2.As shown in Table 2, the GGA administration can suppress diarrhoea that DSS enteritis causes or the morbidity of having blood in stool, and also has and suppress to lose weight or effect that intestinal tube shrinks.
Hence one can see that, and GGA is very effective to the DSS enteritis as the inflammatory bowel disease model.
Table 1, GGA administration are to the histological score of the inhibition effect of DSS enteritis
Tissue injury The scope of inflammation
Contrast 3.7±1.5 2.7±0.6
GGA 1.3±0.6 1.3±0.6
Table 2, GGA administration are to the inhibition effect of DSS enteritis
Diarrhea disease percentage (%) The sickness rate (%) of having blood in stool (%) loses weight The length of intestinal (cm)
Contrast 100 100 86.4±4.4 5.1±0.1
GGA 33 67 93.8±5.1 6.7±0.4
Above result has confirmed that the GGA oral administration suppresses inductive diarrhoea of DSS and clinical disease symptoms such as hemorrhage of rectum and body weight loss.And, the GGA oral administration, the expression of inducing HSP70 in large intestine suppresses the activity level of MPO and the generation of proinflammatory cytokine.In addition, induce in the colitis at the TNBS different with DSS enteritis model, the effect of GGA also obtains confirming.Known to mice, acute TNBS induces colitis to induce enteritis serious than DSS.The survival rate of the mice that the high dose administration of TNBS is handled can improve by the oral administration of GGA, and body weight loss is also owing to GGA and by free burial ground for the destitute inhibition intentionally.By The above results as can be known the high expressed of GGA by HSP70 show anti-inflammatory effect, in the treatment of people IBD effectively.
Utilizability on the industry
According to the present invention, by the GGA administration, in large intestine, induce heat shock protein, to enteropathy, therefore preventing and/or treating effectively of IBD particularly the invention provides and contain GGA as the agent that prevents and/or treats of the enteropathy of active ingredient.

Claims (24)

1, a kind of enteropathy preventive and/or therapeutic agent, its hydrate that contains heat shock protein derivant or its salt or above-claimed cpd is as effective ingredient.
2, enteropathy preventive as claimed in claim 1 and/or therapeutic agent, wherein said heat shock protein derivant is the iso-amylene ketone compounds.
3, enteropathy preventive as claimed in claim 2 and/or therapeutic agent, wherein said iso-amylene ketone compounds are 6,10,14 shown in the formulas (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
4, as claim 1～3 each described enteropathy preventive and/or therapeutic agent, wherein said enteropathy is an inflammatory bowel.
5, enteropathy preventive as claimed in claim 4 and/or therapeutic agent, wherein said inflammatory bowel are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
6, as claim 1～5 each described enteropathy preventive and/or therapeutic agent, wherein said heat shock protein is a heat shock protein 70.
7, a kind of inflammatory bowel preventive and/or therapeutic agent, it is with 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as effective ingredient.
Figure A2004800135740002C2
8, inflammatory bowel preventive as claimed in claim 7 and/or therapeutic agent, wherein said inflammatory bowel are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
9, a kind of prevention method of enteropathy and/or Therapeutic Method, it comprises the step of the hydrate that gives heat shock protein derivant or its salt or above-claimed cpd.
10, the prevention method of enteropathy as claimed in claim 9 and/or Therapeutic Method, wherein said heat shock protein derivant is the iso-amylene ketone compounds.
11, the prevention method of enteropathy as claimed in claim 10 and/or Therapeutic Method, wherein said iso-amylene ketone compounds are 6,10,14 shown in the formulas (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
12, as the prevention method and/or the Therapeutic Method of each described enteropathy of claim 9～11, wherein said enteropathy is an inflammatory bowel.
13, prevention method as claimed in claim 12 and/or Therapeutic Method, wherein said inflammatory bowel are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
14, as the prevention method and/or the Therapeutic Method of each described enteropathy of claim 9～13, wherein said heat shock protein is a heat shock protein 70.
15, a kind of prevention method of inflammatory bowel and/or Therapeutic Method, it is with 6,10,14 shown in the formula (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as effective ingredient.
16, prevention method as claimed in claim 15 and/or Therapeutic Method, wherein said inflammatory bowel are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
17, the hydrate of heat shock protein derivant or its salt or above-claimed cpd contains the hydrate of heat shock protein derivant or its salt or above-claimed cpd as the enteropathy preventive of effective ingredient and/or the purposes in the therapeutic agent in preparation.
18, purposes as claimed in claim 17, wherein said heat shock protein derivant is the iso-amylene ketone compounds.
19, purposes as claimed in claim 18, wherein said iso-amylene ketone compounds are 6,10,14 shown in the formulas (I), 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone.
20, as each described purposes of claim 17～19, wherein said enteropathy is an inflammatory bowel.
21, purposes as claimed in claim 20, wherein said diseases associated with inflammation are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
22, as each described purposes of claim 17～21, wherein said heat shock protein is a heat shock protein 70.
23, the hydrate of heat shock protein derivant or its salt or above-claimed cpd contains 6,10,14 shown in the formula (I) in preparation, 18-tetramethyl-5,9,13,17-19 carbon tetraene-2-ketone is as the preventive of the inflammatory bowel of effective ingredient and/or the purposes in the therapeutic agent.
24, purposes as claimed in claim 23, wherein said inflammatory bowel are the diseases that is selected from clone disease, ulcerative colitis, intestinal type behcets disease and ischemic enteritis.
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