CN103200954B - For treating ulcerative colitis (UC) and nAChR α 7 agonist of Crohn disease (CD) and nAChR α 7 antagonist - Google Patents

Abstract

Disclose the agonist of nAChR α 7 for treating ulcerative colitis (UC) and Crohn disease (CD) and the antagonist of nAChR α 7 and they are as the purposes of the therapeutic agent in order to treatment and management inflammatory bowel (IBD) such as Crohn disease (Crohn ' s disease, CD) and ulcerative colitis (UC).

Description

Exciting for treating the nAChR α 7 of ulcerative colitis (UC) and Crohn disease (CD) Agent and nAChR α 7 antagonist

Cross-Reference to Related Applications

This application claims on June 25th, 2010 submit to the rights and interests that U.S. Provisional Application No. is 61/358,481, its This is fully incorporated by quoting.

Background

Inflammatory bowel (IBD), includes but not limited to Crohn disease (CD) and ulcerative colitis (UC), is chronic intestinal disease Suffering from, it only just affects about 3,400,000 people in western countries and causes huge ill and health care to spend.In the past Decades, scientist and clinician are puzzled by following observed result: the tobacco product of smoking is useful to the process of UC Affect and the process of CD is had injurious effects.But, the effect in the process of UC and CD of smoking and nicotine does not has quilt It is best understood by.

General introduction

In some respects, disclosure theme provides and treats inflammatory bowel in the curee having corresponding treatment to need (IBD) method, described method includes effectively regulating the amount of nAChR α 7 activity of at least one cell in described curee Described curee is used nAChR α 7 (nAChR α 7) antagonist, at least one cell described whereby The IBD of curee described in the adjustment for the treatment of of nAChR α 7 activity.In particular aspect, IBD includes Crohn disease (CD).

In other respects, disclosure theme provides and treats inflammatory bowel in the curee having corresponding treatment to need (IBD) method, described method includes effectively regulating the amount of nAChR α 7 activity of at least one cell in described curee Curee uses nAChR α 7 agonist, and at least one cell described whereby, the adjustment for the treatment of of nAChR α 7 activity is treated The IBD of person.In particular aspect, IBD includes ulcerative colitis (UC).

At other aspect, disclosure theme provides the nAChR α 7 for regulating at least one immune cell type to live Property method, described method include making at least one immune cell type with effectively to regulate at least one immune cell type NAChR α 7 antagonist of amount of nAChR α 7 activity or nAChR α 7 agonist exposure.

In other respects, disclosure theme provides for reducing development knot in the curee have chronic inflammatory disorders of gastrointestinal tract The method of the risk of rectal cancer, described method includes effectively to regulate nAChR α 7 in one or more gastrointestinal tract immunocytes alive Property amount described curee is used nAChR α 7 antagonist or nAChR α 7 agonist, whereby regulation nAChR α 7 activity change Inflammatory reaction in one or more gastrointestinal tract immunocytes, therefore reduces the risk of development colorectal cancer.

In some respects, disclosure theme provides for identifying regulation at least one cell expressing nAChR α 7 The compound of nAChR α 7 activity or the method for agent, described method includes: (i) makes at least one cell and the time of expression nAChR α 7 Select compound or agent contact；(ii) at least one cell of the expression nAChR α 7 contacted with described candidate compound or agent is determined The activity of middle nAChR α 7；(iii) at least the one of the expression nAChR α 7 not contacted with described candidate compound or medicament is determined Plant the activity of nAChR α 7 in compared with control cells；(iv) at least one cell that contrast contacts with described candidate compound or agent The activity of nAChR α 7 in the activity of nAChR α 7 and at least one compared with control cells；Wherein contacting with described candidate compound or agent At least one cell in nAChR α 7 activity with the differential identification of nAChR α 7 activity at least one compared with control cells extremely Few a kind of cell regulates candidate compound or the agent of nAChR α 7 activity.

The most in other respects, disclosure theme provides for prediction suffering from the gene by nAChR α 7 or albumen table The curee of the IBD that the abnormal level reached causes uses the method for the therapeutic effect of nAChR α 7 Expression modulation agent, wherein NAChR α 7 Expression modulation agent is nAChR α 7 antagonist or nAChR α 7 agonist, and described method includes: (a) measures and using The tissue of curee before nAChR α 7 regulator or the gene of nAChR α 7 in cell or protein expression level；B () is with effectively In the tissue of change curee or cell, curee is used nAChR α 7 by the amount of nAChR α 7 gene or protein expression level Regulator；C () is measured and is come from the nAChR α's 7 that uses in the tissue of the curee after nAChR α 7 regulator or cell Gene or protein expression level；(d) determine and use the gene of nAChR α 7 in the tissue after nAChR α 7 regulator or cell Or protein expression level is compared to use the tissue before nAChR α 7 regulator or the gene in cell or protein expression level Change；Wherein the change prediction nAChR α 7 of nAChR α 7 gene in tissue or cell or protein expression level expresses and adjusts The therapeutic effect of the joint agent curee to suffering from IBD.

In some respects, disclosure theme provides pharmaceutical composition, and it includes that the function effectively regulating nAChR α 7 is with treatment Or prevention has the nAChR α 7 (nAChR of amount of inflammatory bowel (IBD) of curee that corresponding treatment needs α 7) specific agonist or specific antagonists.

Some aspect of disclosure theme is the most stated herein above, and it is mentioned whole or in part by disclosure theme；When With when hereinafter the optimal appended embodiment described and accompanying drawing connect, other aspects will carry out along with description and become bright Aobvious.

Accompanying drawing is sketched

Have thus described disclosure theme with general terms, referring now to accompanying drawing, accompanying drawing is not drawn necessarily to scale, And wherein:

Fig. 1 shows the representational hierarchical clustering (hierarchical clustering) from microarray spectrum, display Expression patterns different in IBD.Every a line represents gene, and independent biological sample is shown in each list.Gene in immunocyte The cluster analysis of expression map shows diversity mode, and it is clustered into four groups of following display: normal (N), CD, UC, and just Often (N).Shadow representation is lowered or overexpression, and black represents similar expression；

Fig. 2 shows the differentiation potentiality of 10 genes identified by DFA.DFA is used for selecting maximum discrimination variable (maximal discriminatory variable), and UC, CD be grouped into 3 different positions with normal healthy controls；

Fig. 3 shows that the nAChR α 7 that identified by gene microarray and confirmed by QRT-PCR is at UC, CD and unaffected right Differential gene expression pattern according to.Zuo Tu: microarray data: from standardized nAChR α 7 expression of immunocyte It is described in UC and CD the differential gene expression pattern of uniqueness.N: neutrophil cell；P:PMBC；Ctrl: comparison.Right figure: The QRT-PCR checking to microarray results.QRT-PCR confirms the gene expression results of nAChR α 7 and from the five (5) of PBMC The gene expression results (data do not show) of other genes selected individual；

Fig. 4 proves that nAChR α 7 mediates different immunomodulating collection of illustrative plates in CD is relative to the lymphocyte of UC.Show From the lymphocyte of the conversion processed in UC and CD compared to the cytokine profiles in the supernatant of untreated cell Ratio (at least three be similar in experiment).Process include nicotine [2 μMs (+) and 20 μMs (++)] and add nicotine Before use selective antagonist alpha-bungarotoxin (α-bungarotoxin) [2nM (+) and 20nM (++)] pretreatment.A schemes: Cytokine IL-10 and cytokine/chemotactic factor IL-8.B schemes: chemotactic factor.For all unaffected comparisons, add The change (not shown) of the cytokine/chemotactic factor of nicotine or alpha-bungarotoxin itself non-limiting.Nic: nicotine. Antg: selective antagonist alpha-bungarotoxin；

Fig. 5 shows that nicotine and nAChR α 7 specific agonist are to " CD sample " TNBS colitis and " UC sample " DSS colitis The reverse effect of process.These IBD mouse models are considered as " CD sample " or " UC sample " not only due to their disease phenotype Similar with the mankind CD or UC respectively, and because their cytokine profiles is similar with the mankind CD or UC, especially The chronic model of TNBS or DSS model, foregoing (Alex et al., 2009 IBD, 15 (3), 341-352).These experiments Property mouse colitis model produces (Alex et al., 2009 IBD, 15 (3), 341-352) foregoing.Ethanol: TNBS's is right According to；Nic: nicotine；α 7Ag:nAChR α 7 agonist PNU282987 (PNU)；

Fig. 6 shows that nAChR α 7 agonist is not only preventative for UC sample DSS colitis, and is curative. In acute model, mice processes with or without nAChR α 7 anti-depressant medications, continues 5 days.Then give DSS, continue 7 days.Comment Valency Clinical Activity scoring (clinical activity score).In chronic model, mice is given DSS, continues 7 days, Then the water of seven days is given.The circulation of these DSS-water continues to repeat 3 times again.At the end of the 4th circulation, even without Under DSS, mice develops into chronic colitis, and it has the Clinical Activity of about 3 to mark.At this moment, nAChR α 7 agonist is given Give one group of mice (one set of mice) and follow the tracks of all mices 11 days.In acute model and chronic model, disease (colitis) substantially disappears and mice seems do not have difference with control mice (without DSS process).Arrow indicates chronic The beginning (the 53rd day) that in model, nAChR α 7 agonist processes；

Fig. 7 shows that the pretreatment of MLA medicine (nAChR α 7 specific antagonists) can effectively prevent mice to develop CD sample TNBS colitis.Continued once a day by lumbar injection with PBS (comparison) or nicotine, PNU (α 7 agonist) or MLA medicine 5 days pretreatment C57BL/6 mices.Then TNBS (in 50% ethanol) or only 50% ethanol is used to send out as comparison inducing mouse Exhibition colitis, continues 7 days.Every day monitors the disease activity of all mices.Using MLA (α 7 antagonist+TNBS) pretreatment In the mice that TNBS processes, disease (colitis) substantially disappears.The mice that these MLA process seems and compareing without TNBS Mice (ethanol) does not has difference.On the contrary, use nicotine (nicotine+TNBS) or the mice of PNU (α 7 agonist+TNBS) pretreatment, Compared with only TNBS process (TNBS), display does not all have therapeutic benefit, even deteriorating condition movable.

Fig. 8 shows use nAChR α 7 specific antagonists (Antg；MLA) effectively reverse is processedChronicCD sample TNBS induces The disease process of colitis.The induction of chronic TNBS colitis model as previously mentioned (Alex et al., 2009IBD, 15 (3), 341-352).The DAI of the colitis mice that antagonist processes is similar to control mice (using ethanol (E-OH) to process).nAChRα 7 agonist (Ag；PNU) show without significance effect with nicotine.Use ANOVA and Tukey inspection (Tukey ' s test) carry out Statistical analysis；P ＜ 0.05.N >=12 often group；

Fig. 9 shows the colitis reversing TNBS induction that processes using nAChR α 7 antagonist: (A) comes from and have place The colon of the mice of the colitis of the chronic TNBS induction of reason is shorter, inflammation, and is filled with courageous and upright and soft feces； (A-c), and coming from the colon of colitis mice that antagonist processes is the most normal outward appearance (A-d), with control mice (A- A&b) compare almost without difference；(B) the TNBS mice that the histologic analysis display antagonist using H&E dyeing to carry out processes has Normal mucous membrane of colon and almost without inflammatory signs (B-c), as seen in the mice processed without TNBS (B-b).Show the representative of at least ten independent experiment；

Figure 10 shows that DSS or TNBS of nAChR α 7 defective mice is all lured by nAChR α 7 specific agonist and antagonist The colitis led, without any therapeutic effect, shows the specificity of these medicines.Wild type (WT) nAChR α 7 defective (nAChR α 7 knocks out；α 7KO) mice, all with C57B/6 as background, use DSS (A) or TNBS (B) induction development colitis.Although it is out of office The colitis of agonist (PNU) and antagonist (MLA) the most effectively prevention DSS and TNBS induction in raw type mice, at nAChR α In 7KO mice they to any colitis all entirely without showing any effect.These data further illustrate these Agonist and the antagonist target-specific to nAChR α 7；Significant difference in measured value by ANOVA and Tukey inspection with 5% significance level evaluation.N >=6 often group；

Figure 11 shows that the mice that experienced by vagotomy has completely lost nAChR α 7 agonist to colitis development Protective effect: (A) is before and after vagotomy: based on before report (Verma-Gandhu et al., Gut 2007:56:358-364) on the basis of amendment scheme carry out vagotomy.Briefly, C57B/6 mice (7 Week old) left vagus nerve branch (the about 5-10mm of gastroesophageal junction (marking with white arrow)；With double-headed arrow table Show) microscope amplify under (10 ×) by use electrocautery cut off (S, stomach；E, esophagus).Operation opening uses absorbable stitching Suturing with thread management.Sham-operation includes step (only exposing) same in addition to Vagotomy.Experienced by vagus nerve to cut The mice time of 7 days of disconnected art recovers.Experienced by all 30 mices 100% of operation (vagotomy and sham-operation) Survive and there is no complication, and recover to normally.Then give mice nAChR α 7 agonist with IP, continue 5 days, laggard Row DSS process (7 days) is with inducing colitis；(B) mice (vagotomy) that experienced by vagotomy does not has Represent nAChR α 7 agonist any protectiveness effect to colitis, and experienced by mice (sham-operation) reaction of sham-operation with The mice not experiencing any operation reacts the best (see Fig. 5 and 6).The significant difference of measured value uses Wilcoxon to be lost Evaluate with inspection (Wilcoxon rank-sums test) (P ＜ 0.05).N >=5 often group；

Figure 12 shows that nAChR α 7 agonist has therapeutic effect for GPX 1/2DKO colitis.Lack two glutathion The mice [Gpx1/2 is double knocks out (DKO) mice] of peroxidase (GPX) Gpx1 and Gpx2 is at C57BL/6 and 129S1/ of mixing The tendency of ileocolitis is had under SvJ (B6.129) genetic background.Evaluation Clinical Activity is marked.Comparison (comes from mixing C57BL/6 and 129S1/SvJ (B6.129) genetic background) do not develop serious colitis, and GPX1/2 DKO mice is found Development Clinical Activity scoring is about the severe colitis of 9.The GPX1/2 DKO mice processed with nAChR α 7 agonist In, the development of disease (colitis) substantially reduces.But use nAChR α 7 antagonist or the GPX1/2 DKO mice of saline treatment Colitis score does not represent significance change；

Figure 13 show discrimination function analysis (Discriminant Functional Analysis) (DFA) identification of cell because of The variable of son/trend factor, it can sufficiently distinguish a disease type and other diseases type and differentiation treatment response Person (therapeutic responder) with compare.DSS induction and TNBS induction colitis form different groups, its On collection of illustrative plates with to photograph away from, and each other away from.On the other hand, with the mice of the DSS induction of agonist process with short of money The mice of the TNBS induction that anti-agent processes on collection of illustrative plates very close in comparison with each other in effective process of respective medicine After, cytokine profiles clearly shifts to normal condition from morbid state；With

Figure 14 shows that forced swim test display nAChR α 7 agonist (PNU) does not has measurable effect to mice behavior. By forcing mice swimming to test for 6 minutes in hydrostatic column.Mice is divided into 4 groups: (a) compares；B () accepts IP and swashs The mice of dynamic agent；C mice that () induces with DSS；(d) IP agonist the mice (n=6 often group) with DSS induction are accepted.? In 25 DEG C of containers having 13cm water, the total duration of motionless (immobility) is scored.Motionless it is defined as when mice is at water Middle stopping struggles and keeps floating static, only makes and maintains its head during necessary action on the water surface.0th day and the 7th day table Show the sky that DSS processes.

Describe in detail

Disclosure theme will be described now the most completely, shows disclosure theme in the accompanying drawings Some but not all embodiment.Throughout similar numeral refers to similar key element.Disclosure theme can be with the most not The embodiment that should not be construed as being limited to state herein it is carried out with form；On the contrary, it is provided that these embodiments so that this The open satisfied legal requiremnt being suitable for.Really, the disclosure theme of the teaching presented in described above and relevant drawings is benefited from One of ordinary skill in the art will recognize that a lot of amendments and other embodiments of the disclosure theme stated herein.Therefore, manage Solving, disclosure theme is not only restricted to disclosed specific embodiment, and is intended to include amendment and other embodiments In the range of attached claim.

I. it is used for treating ulcerative colitis (UC) and nAChR α 7 agonist of Crohn disease (CD) and nAChR α 7 antagonism Agent

In some embodiments, disclosure theme identifies nAChR α 7 (nAChR α 7) is regulation The smoking (nicotine) the main regulatory factor to the CD opposite effect compared to UC.Therefore, nAChR α 7 is identified by disclosure theme New therapeutic target for IBD.Disclosure theme provides specific agonist and the antagonist of nAChR α 7, and they are as being used for The purposes of the therapeutic agent for the treatment of and management UC and CD.

A. the effect of smoking in IBD (nicotine)

Inflammatory bowel (IBD) is a kind of popular chronic progressive external polyfactorial gastrointestinal (GI) road inflammatory condition, its performance In two different common solid: ulcerative colitis (UC) and Crohn disease (CD).Xavier R.J., Podolsky D.K, " Unravelling the pathopenesis of inflammatory bowel disease, " Nature 488: 427-434(2007).Phrase " inflammatory bowel " or " IBD " are intended to include two kinds to affect gastrointestinal chronic as used here Disease, referred to as Crohn disease (CD) and ulcerative colitis (UC).Term Crohn disease and ulcerative colitis be meant that by The generally accepted definition of professional and technical personnel of medical domain.

Only just having more than 1,000,000 patients being diagnosed in the U.S., IBD has the prevalence of about 0.2% in the population of west And there is substantial amounts of ill and health care cost.Loftus E.V., Jr., " DEG C linical epidemiology of Inflammatory bowel disease:Incidence, prevalence, and environmental Influences, " Gastroenterology 126:1504-1517 (2004).The major complications of the IBD of colon is to increase The risk of development colorectal cancer (CRC), mainly due to chronic GI inflammation.Shanahan F., " Review article: Colitis-associated cancer--time for new strategies 1, " Aliment Pharmacol Ther 18 Suppl 2:6-9 (2003)；Campbell B.J., Yu L.G., Rhodes J.M., " Altered glycosylation In inflammatory bowel disease:a possible role in cancer development, " Glycoconj J 18:851-858 (2001).It is interesting that smoking has wholesome effect to the process of UC, and CD is entered Journey has injurious effects.Birrenbach T., Bocker U., " Inflammatory bowel disease and Smoking:a review of epidemiology, pathophysiology, and therapeutic Implication, " Inflamm Bowel Dis 10:848-859 (2004)；Rubin D.T., Hanauer S.B., " Smoking and inflammatory bowel disease, " Eur J Gastroenterol Hepatol 12:855- 862(2000)；Thomas G.A., Rhodes J., Ingram J.R., " Mechanisms of disease:nicotine-- A review of its actions in the context of gastrointestinal disease, " Nat Clin Pract Gastroenterol Hepatol 2:536-544 (2005)；With Ingram J.R., Rhodes J., Evans B.K., Thomas G.A., " Preliminary observations of oral nicotine therapy for Inflammatory bowel disease:an open-label phase I-II study of tolerance, " Inflamm Bowel Dis 11:1092-1096 (2005).

Ulcerative colitis (UC) has been identified as mainly non smoker and/or the disease of Ex-smoker, and, to the greatest extent The starting protection and improve the process of UC of pipe smoking, but smoking terminate deteriorating the process of UC.Although nicotine, the activity in smoking Part, illustrates the notable clinical efficacy to UC in clinical trial as treatment, but its application is become by considerable toleration Change and side effect is limited.On the contrary, the CD patient of about 72% is smoker, and, although smoking adds the risk of CD also And the process being degrading CD reaches more serious form (structuring or fistula), but the termination of smoking significantly improves CD.Birrenbach T., Bocker U., " Inflammatory bowel disease and smoking:a review Of epidemiology, pathophysiology, and therapeutic implication, " Inflamm Bowel Dis 10:848-859 (2004).The molecular mechanism of the smoking opposite effect on CD and UC still keeps mysterious recent decades.

B. nAChR (nAChR)

Nicotine has been elucidated with by its triggered inflammatory cascade reaction to nAChR, nAChR be part gate from Subchannel family, this family is by multiple homology pentamer (homopentameric) or allos pentamer (heteropentameric) receptor subunit is formed, and it is made up of and god two subfamily 9 α (α 2-α 10) and 3 β (β 2-β 4) Expression is all had in system and non-nervous tissue's (such as immunocyte and enterocyte).Gaimarri A., Moretti M., Riganti L., Zanardi A., Clementi F., Gotti C., " Regulation of neuronal nicotinic Receptor traffic and expression, " Brain Res Rev 55:134-143 (2007).Nicotine is to UC phase Ratio is the main pharmaceutical component of the opposite effect in CD.Thomas G.A., Rhodes J., Ingram J.R., " Mechanisms of disease:nicotine--a review of its action in the context of Gastrointestinal disease, " Nat Clin Prac Gastroenterol Hepatol 2:536-544 (2005)。

Nicotine clinical efficacy in UC treats shows that NAChR is the most possible target for the treatment of based on nicotine.But Its clinical practice, is limited by change and the side effect of considerable toleration, and this is likely due to the nicotine part to nAChR Specificity.Recent studies suggest that, nicotine serves as antiinflammatory especially by the generation suppressing proinflammatory cytokine in macrophage Agent.Gallowitsch-Puerta M., Tracey K.J., " Immunologic role of the cholinergic Anti-inflammatory pathway and the nicotinic acetylcholine alpha 7 receptor, " Ann N Y Acad Sci 1062:209-219 (2005).This phenomenon is defined as " cholinergic anti-inflammatory pathway ", wherein neural Peptide is as the molecular basis of nerve immunity axle, and this path provides antiinflammatory to feed back by autonomic nervous system (ANS) to immune system. Tracey K.J., " The inflammatory reflex, " Nature 420:853-859 (2002).This path is known logical Cross efferent vagus nerve from neural cholinergic inflammatory signals transmission and to be regulated by the specificity of nAChR；Particularly immunity is thin The nAChR α 7 of the upper expression of born of the same parents.Wang H., Yu M., Ochani M., Amelia C.A., Tanovic M., Susarla S., Li J.H., Wang H., Yang H., Ulloa L., Al-Abed Y., Czura C.J., Tracey K.J., " Nicotinic acetylcholine receptor alpha7 subunit is an essential regulator of Inflammation, " Nature 421:384-388 (2003)；De Jonge W.J., van der Zanden E.P., The F.O., Bijlsma M.F., van Westerloo D.J., Bennink R.J., Berthoud H.R., Uematsu S., Akira S., van den Wijngaard R.M., Boeckxstaens G. E., " Stimulation of the vagus nerve attenuates macrophage activation by activating the Jak2-STAT3 signaling Pathway, " Nat Immunol 6:844-851 (2005)；Floto R.A., Smith K.G., " The vagus nerve, Macrophages, and nicotine, " Lancet 361:1069-1070 (2003)；Ghia J.E., Blennerhassett P., Kumar-Ondiveeran H., Verdu E.F., Collins S.M., " The vagus nerve:a tonic inhibitory influence associated with inflammatory bowel disease in a murine Model, " Gastroenterology 131:1122-1130 (2006).

NAChR α 7 is the nAChR being widely studied most.Mazurov A., Hauser T., Miller C.H., " Selective alpha7 nicotinic acetylcholine receptor ligands, " Curr Med Chem 13: 1567-1584(2006).Add recently and much exist in nAChR α 7 part and the interest of their purposes, and these agent Carry out clinical trial (but not being for IBD).These are in progress to provide identifies and designs/synthesize dividing of nAChR α 7 specific drug Subbase plinth.

C. representative embodiment

Disclosure theme identifies nAChR α 7 as regulation smoking (i.e. nicotine exposure) to the CD contrary effect relative to UC The main regulatory factors of fruit.First, the gene expression atlas of the lymphocyte and neutrophil cell by coming from IBD patient Find nAChR α 7 hypotype (nAChR α 7), compared to unaffected in contrast, at Crohn disease (CD) Notable overexpression in patient, but lowered in ulcerative colitis (UC) patient.Undesirably by any concrete reason Opinion constraint, it is assumed that smoking may cause reverse effect to UC compared to CD by nAChR α 7.For checking this it is assumed that use non- (including PNU 282987 (PNU), GTS-21, and MLA, as hereafter determined for ligands specific (nicotine) and some ligands specifics Justice) to detect them to UC sample DSS colitis in mice IBD model and the effect of CD sample TNBS colitis, these mice moulds Type is the animal model commonly used most in the preclinical study of the curative drug of essentially all IBD.

In some embodiments, disclosure theme illustrates nAChR α 7 agonist PNU 282987 (N-[(3R)-1-nitrogen Miscellaneous dicyclo [2.2.2] oct-3-yl]-4-chlorobenzamide) and 3-(2,4-dimethoxybenzyliden) Anabaseine (3-(2,4- Dimethoxybenzylidene) anabaseine, GTS-21, the most also referred to as DMXB-A) it is height to UC sample colitis Effective curative, but deteriorate CD sample colitis, this observed result and smoking are similar to the mankind UC effect compared to CD.

On the other hand, in other embodiments, disclosure theme illustrates nAChR α 7 antagonist methyllycaconitine (MLA) it is to the CD highly effective therapeutic agent of sample colitis, but to UC sample colitis to no effect.Reiterate, it is undesirable to be subject to Any concrete theoretical constraint, it is believed that these agonist and antagonist are by regulation anti-inflammatory cytokines and proinflammatory cytokine Effect and play a role.Such as, agonist can stimulate anti-inflammatory cytokines, and suppresses proinflammatory cytokine.And, these public affairs Opening theme and illustrating the effect of PNU and MLA is that specificity passes through nAChR α 7, because its effect produces IBD's being induced NAChR α 7 deficient mice is not observed.Therefore, disclosure theme not only characterizes the smoking effect to IBD, and identifies The nAChR α 7 molecular target as the Results for CD and UC.

More particularly, in some embodiments, disclosure theme provides has what corresponding treatment needed to be subject to for treatment The method of the inflammatory bowel (IBD) of therapist, described method includes effectively regulating at least one cell in described curee In nAChR α 7 activity amount curee is used nAChR α 7 (nAChR α 7) antagonist, the most extremely The IBD of the adjustment for the treatment of curee of nAChR α 7 activity in few a kind of cell.In some embodiments, IBD includes Crow Grace is sick (CD).

In other embodiments, disclosure theme provides the inflammatory of the curee having corresponding treatment to need for treatment The method of enteropathy (IBD), described method includes the nAChR α's 7 effectively to regulate in described curee at least one cell Curee is used nAChR α 7 (nAChR α 7) agonist, whereby at least one cell by the amount of activity The IBD of adjustment for the treatment of curee of nAChR α 7 activity.In some embodiments, IBD includes ulcerative colitis (UC)。

" effective dose " of term activating agent guides out the necessary amount of desired biologically.As will be general by this area Logical technical staff is understood, and the effective dose of agent may change with various factors, and these factors are whole for the most desired biology Point, the agent being delivered, the composition of pharmaceutical composition, target tissue or cell, like this.More particularly, term " effective dose " refers to Be enough to produce the amount of desired effects, described desired effects such as reduces or improves seriousness, persistent period, process, or disease, disease The outbreak of disease or illness (such as relevant to losing cell function disease, disease or illness) or one or more symptom；In advance The progress of anti-disease, disease or illness；Cause disappearing of disease, disease or illness；Prevent relevant with disease, disease or illness The recurrence of symptom, develop, show effect or be in progress；Or strengthen or improve prevention or the therapeutic effect of another kind treatment.According to the disclosure Method, the scope of the effective dose of compound is it may be that such as from about 0.001mg/kg to about 1000mg/kg, or implements at some In scheme, from about 0.01mg/kg to about 100mg/kg, or in certain embodiments, from about 0.1mg/kg to about 50mg/kg. As those skilled in the art will realize, effective dose is also with changing as follows: the illness for the treatment of, route of administration, excipient Use, the age of curee and sex, and the possibility being used in conjunction with other treatment treatment (such as using other medicaments) Property.It will be appreciated that may be effective for other purposes with compound for arriving the amount of the compound required for expectation biologically Amount different.

As used here, term " agonist " is defined as stimulating or the material of induction nAChR α 7 activity.Term " antagonist " is defined as blocking or the material of suppression nAChR α 7 activity.In some embodiments, nAChR α 7 receptor agonism Agent or antagonist include micromolecular inhibitor, and it includes but not limited to exemplary agonist as provided below and antagonist.But It is that nAChR α 7 activity can regulate by any method known to those skilled in the art, includes but not limited to that biological agent is such as The use of specific antibody, increases gene expression by gene therapy, or by RNAi inhibition of gene expression.

In special embodiment, selective nAChR α 7 agonist is selected from the group consisted of: PNU 282987 (N-[(3R)-1-azabicyclo [2.2.2] oct-3-yl]-4-chlorobenzamide), MEM3454, PH-399733, AR-R1779, SSR180711A (4-bromophenyl Isosorbide-5-Nitrae diazabicyclo (3.2.2) nonane-4-carboxylate, mono-hydrochloric salts (4-bromophenyl Isosorbide-5-Nitrae diazabicyclo (3.2.2) nonane-4-carboxylate, monohydrochloride)), ABT-418 (3-first Base-5-[(2S)-1-methylpyrrolidin-2-yl]-1,2-azoles), methiodide cocaine (cocaine methiodide), 3-2,4- Dimethoxybenzyliden Anabaseine (GTS-21 or DMXB-A), 3-(4-phenol methylene) Anabaseine, 3-(4-methoxy Base benzylidene) Anabaseine, 3-(4-aminobenzene methylene) Anabaseine, 3-(4-hydroxyl-2-methoxybenzylidene) be false Herba Equiseti Hiemalis because of, 3-(4-methoxyl group-2-phenol methylene) Anabaseine, trans-3-cinnamylidene Anabaseine, trans-3-(2- Methoxyl group-cinnamylidene) Anabaseine and trans-3-(4-methoxyl group cinnamylidene) Anabaseine.

In other embodiments, selective nAChR α 7 agonist is selected from the group consisted of: N-[(3R)-1-nitrogen Miscellaneous dicyclo [2.2.2] oct-3-yl]-4-(4-hydroxyphenoxy) Benzoylamide, N-[(3R)-1-azabicyclo [2.2.2] octyl-3- Base]-4-(4-acetylamino phenoxy group) Benzoylamide, N-[(3R)-1-azabicyclo [2.2.2] oct-3-yl]-4-(phenyl sulfur Alkyl phenoxy) Benzoylamide (N-[(3R)-1-azabicyclo [2.2.2] oct-3-y1]-4- (phenylsulfanylphenoxy) benzamide), N-[(3R)-1-azabicyclo [2.2.2] oct-3-yl]-4-(3-chlorobenzene Sulfonyl) Benzoylamide and (1-aza-bicyclo [2.2.2] oct-3-yl) carbamic acid 1-(2-fluorophenyl)-ethyl ester.

The selectivity nAChRa7 agonist being applicable to disclosure theme: PCT/ is described in following patent application US2006/022136；PCT/US1998/17850；PCT/EP2007/005724；WO99/62505；WO99/03859；WO97/ 30998；WO01/36417；WO02/15662；WO02/16355；WO02/16356；WO02/16357；WO02/16358；WO02/ 17358, U.S. Patent Application Publication No. 2006/0166974；No. 2006/0128676；No. 2005/0282823；2005/ No. 0165047；No. 2004/0044026；No. 2004/0229868, and MazurovA., Hauser T., Miller C.H., " Selective alpha7 nicotinic acetylcholine receptor ligands, " Curr Med Chem 13: 1567-1584 (2006), above-mentioned each of which is passed through to quote to be fully incorporated herein.

In certain embodiments, nAChR α 7 agonist selects free nAChR α 7 specific antibody, nAChR α 7 specificity to press down The group that property RNA processed or a combination thereof are formed.

In some embodiments, selectivity nAChR α 7 antagonist includes alpha-bungarotoxin and methyllycaconitine (MLA). It is described in the following documents the example of other selectivity nAChR α 7 antagonist: Mazurov A., Hauser T., Miller C.H., " Selective alpha7 nicotinic acetylcholine receptor ligands, " Curr Med Chem 13:1567-1584 (2006), it is fully incorporated by introducing at this.In certain embodiments, nAChR α 7 antagonist Select the group of free nAChR α 7 specific antibody, nAChR α 7 specific inhibitory RNA or combinations thereof.

In other embodiments, the part of nAChR α 7 receptor includes but not limited to diazabicycloalkane derivatives, such as As described in WO2005/028477；Volution quinine cyclic ethers (spirocyclic quinuclidinic ether) derivant, Such as described in WO2005/066168；The substituted quinuclidine derivatives of bicyclic heterocycle condensed, such as such as United States Patent (USP) Shen Please disclose No. US2005/0137184；No. US2005/0137204；Described in No. US2005/0245531；3-quinuclidine Base amino-substituted biaryl derivant, such as described in WO2005/066166；The connection of 3-quininuclidinyl hetero atom bridge joint Virtue derivant, such as described in WO2005/066167；Tricyclic derivatives substituted with amino, such as such as WO2005/ Described in 077899, above-mentioned each is passed through to quote to be fully incorporated.

Run through specification and claims, given chemical formula or title and should include all tautomers, homology Compound (congener) and optics and stereoisomer, and wherein there is the racemization mixing of this isomer and mixture Thing.

In some embodiments, treatment includes the IBD's of development or the prevention curee preventing the IBD of curee Progress.Term " treatment (treating) " or " treatment (treatment) " and grammer derivant thereof be intended to include morbid state and The treatment of disease, manage, prevent and cure.

Term " curee " refers to organism, tissue or cell.Curee can include the mankind for goals of medicine Curee, the most existing disease of goals of medicine or the diagnosis of disease and/or treatment, or prevention disease or outbreak pre-of disease Anti-property treat, or for medical science, veterinary purposes or grow purpose animal subject.Curee can also include from Specimen material like this is produced in tissue culture, cell cultivation, organ partitioning, stem cell.Suitably animal subject Including mammal and birds.Terms used herein " mammal " includes but not limited to primates, such as people, monkey, ape, With like this；Bovine, such as cattle, bullock (ox), and like this；Sheep class (ovines), such as sheep (sheep) and all Such as this type of；Goat class (caprines), such as goat (goat) and like this；Swine, such as pig, barren sow (hog) and such as This type of；Horse class, such as horse, donkey, zebra and like this；Cat class, including wild and domestic cat；Dog class, including Canis familiaris L.；Lagomorpha, Including rabbit (rabbit), hare (hare) and like this；And Rodents, including mice, rat, and like this.Herein The term " birds " used includes but not limited to, chicken, duck, goose, Carnis Coturnicis japonicae, turkey and pheasant.Preferably, curee is mammals Animal or cells of mamma animals.It is highly preferred that curee is the mankind or human cell.Human subject includes but not limited to, Fetus, neonate, baby, teenager, and adult curee.And, " curee " can include suffering from or just suspecting Suffer from the patient of disease or disease.Therefore, term " curee " and " patient " are used interchangeably herein.Curee is also The cell in laboratory or cell gleanings can be referred to or produce for activity, differentiation, mark, express and survey like this Biological treatment culture in examination.

In some embodiments, curee is Rodents, such as mice, and it has been IBD model by chemical induction.? In specific embodiment, mouse model is following one or more: (a) is exposed to dextran sulfate sodium (DSS) with induction The mice of ulcerative colitis sample disease；B () is exposed to trinitro-benzene-sulfonic acid (TNBS) to induce the little of Crohn disease sample disease Mus；C () induction produces nAChR α 7 deficient mice of colitis；Or (d) lacks two kinds of glutathion peroxidase (GPX) Gpx1 and Gpx2, [Gpx1/2-double knocks out (DKO) mice], and and then develop the mice of ileocolitis.

In other embodiments, method of disclosure is for eukaryotic cell, preferred mammal cell, and more preferably people Class cell.Cell or cell line can be passed through commercially-available or can separate from mammalian tissues.It is suitable for the disclosure The cell of method is also used as the part existence of tissue or organ prepared product.Cell be not limited to can from rat, cat, horse, Mice, hamster, chicken, sheep, goat, pig, cattle, rabbit, non-human primates and the mankind obtain.

In other embodiments, disclosure theme provides for regulating at least one immune cell type The method of nAChR α 7 activity, described method includes making at least one immune cell type thin with at least one immunity of effectively regulation NAChR α 7 antagonist of the amount of nAChR α 7 activity in born of the same parents' type or nAChR α 7 agonist exposure.

As used here, phrase " immunocyte " is intended to mean that of function in mammalian immune reacts A little cells, immunoreation includes cell-mediated immunoreation and humoral immune reaction.This phrase includes but not limited to, lymph is thin Born of the same parents, antigen-specific cellular poison T lymphocyte, neutrophil cell, peripheral blood lymphocytes (PBMC), macrophage and from Natural killer cell.

The most in other embodiments, disclosure theme provides for reducing the curee having chronic inflammatory disorders of gastrointestinal tract The method of the risk of development colorectal cancer, described method includes effectively regulating in one or more gastrointestinal tract immunocytes Curee is used nAChR α 7 antagonist or nAChR α 7 agonist, whereby regulation nAChR α 7 activity by the amount of nAChR α 7 activity Change the inflammatory reaction in one or more gastrointestinal tract immunocytes, therefore reduce the risk of development colorectal cancer.

In further embodiment, disclosure theme provide for identify regulation express nAChR α 7 at least one is thin The compound of nAChR α 7 activity or the method for agent in born of the same parents, described method includes: (i) makes at least one cell of expression nAChR α 7 Contact with candidate compound or agent；(ii) at least one of the expression nAChR α 7 that contacts with described candidate compound or agent is determined The activity of the nAChR α 7 in cell；(iii) the expression nAChR α 7 not contacted is determined with described candidate compound or agent extremely The activity of nAChR α 7 in few a kind of compared with control cells；(iv) at least one contacted with described candidate compound or agent is contrasted thin The activity of nAChR α 7 and the activity of nAChR α 7 at least one compared with control cells in born of the same parents；Wherein with described candidate compound or agent NAChR α 7 activity at least one cell of contact and the differential identification of nAChR α 7 activity at least one compared with control cells Regulate candidate compound or the agent of nAChR α 7 activity at least one cell.

The most in further embodiment, disclosure theme provide for prediction to suffer from the gene by nAChR α 7 or The curee of the IBD that the abnormal level of protein expression causes uses the method for the therapeutic effect of nAChR α 7 Expression modulation agent, its Described in nAChR α 7 Expression modulation agent be nAChR α 7 antagonist or nAChR α 7 agonist, described method includes: (a) measure exist Use gene or the protein expression level of nAChR α 7 in the tissue of curee before nAChR α 7 regulator or cell；(b) with In effective tissue changing curee or cell, curee is used by the amount of nAChR α 7 gene or protein expression level NAChR α 7 regulator；C () is measured and is come from tissue or the cell of using the curee after nAChR α 7 regulator The gene of nAChR α 7 or protein expression level；(d) determine and use nAChR in the tissue after nAChR α 7 regulator or cell The gene of α 7 or protein expression level are compared to use the tissue before nAChR α 7 regulator or the gene in cell or albumen The change of expression；The wherein change prediction nAChR of nAChR α 7 gene in tissue or cell or protein expression level The therapeutic effect of the α 7 Expression modulation agent curee to suffering from IBD.

In some embodiments, in the curee suffer from IBD, nAChR α 7 is overexpressed to abnormal level, And the reduction prediction of gene or protein expression level is to the positive reaction using nAChR α 7 regulator.In other embodiments, exist Suffer from the expression of nAChR α 7 in the curee of IBD and be adjusted downward to abnormal level, and the increase prediction of gene or protein level To the positive reaction using nAChR α 7 regulator.

In some embodiments, using one or more methods to determine the change of gene expression dose, described method is selected Free RNA blotting, RT-PCR, real-time RT-PCR, in situ hybridization and the group of microarray composition.In other embodiments, Changing of protein expression level uses one or more methods selected from the group consisted of to determine: western blotting, ELISA, mass spectrum, immunohistochemistry and protein array.

Therefore, in some embodiments, method of disclosure may be used for diagnosis, for prognosis or morbid state or disease Monitoring.As used here, term " diagnoses " process referring to predictability, wherein have rated disease, illness or other medical science The existence of disease, do not exist, seriousness or treatment process.For purpose herein, diagnosis also includes for determining therapeutic outcome The process of predictability.Being similar to, term " diagnoses " and refers to determine whether curee represents the one or more of disease or disease Feature.Term " diagnoses " presence or absence including establishing the target of such as target antigen or binding reagents, or establishes or with other Mode determines one or more features of disease or disease, including type, grade, stage or analogue.The most as used herein , term " diagnoses " can be to include the form distinguishing disease and another form.Term " diagnoses " and includes initial diagnosis Or detection, prognosis is, and disease or the monitoring of disease.It addition, the such as " prison of the term in " disease or the monitoring of progression " Control " refer to ongoing to from having or the diagnosis of sample that the doubtful curee with disease or disease obtains.Term is " pre- Afterwards " and derivative form refers to determine or predictive disease or the process of disease.The process of disease or disease can be such as based on life Expection or quality of life determine.In the case of " prognosis " includes accepting treatment or not accepting treatment, disease or the time of disease The determination of process.In the case of considering treatment, prognosis includes the curative effect determined for disease or disease.The most as used herein , term " risk " refers to predictability process, wherein evaluates the probability of particular result.

Therefore, disclosure theme includes monitoring nAChR α 7 gene expression and/or protein level as IBD patient treatment The prognostic of reaction is measured.The effect to nAChR α 7 gene expression and/or protein level in response to therapeutic agent can provide use In the means determining whether patient responds treatment.In the case of nAChR α 7 overexpression, such as Crohn disease (CD) Patient, the reduction of genes/proteins expression should indicate the positive reaction for treatment.In the case of nAChR α 7 lowers, example Such as the patient of ulcerative colitis, the increase of the expression of genes/proteins should indicate the positive reaction for treatment.Can make The method completing by any method well known by persons skilled in the art to detect gene expression dose, includes but not limited to the most firm Those disclosed method.These methods can use the RNA in any tissue sample interested or albumen, and such as IBD organizes Biopsy, surgery specimen that is fresh or that achieve, the circulating cells being present in blood, serum, urine, and cell line.

II. pharmaceutical composition, medicine box and use

On the other hand, present disclose provides pharmaceutical composition, it includes mixing with pharmaceutically acceptable excipient Independent or with one or more extra therapeutic combinations one or more nAChR α 7 antagonisies and/or nAChR α 7 are exciting Agent.It will be appreciated by persons skilled in the art that pharmaceutical composition includes the pharmaceutically acceptable salt of above-claimed cpd.

More particularly, in some embodiments, disclosure theme provides pharmaceutical composition, and it includes effectively regulating NAChR α 7 function has the nicotine type nicotinic second of the amount of the inflammatory bowel (IBD) of curee that corresponding treatment needs with treatment or prevention The specific agonist of acetylcholine receptor α 7 (nAChR α 7) or specific antagonists.In some embodiments, described medicine group Compound include pharmaceutically acceptable carrier, adjuvant or the excipient nontoxic with one or more together with treatment have NAChR α 7 agonist/antagonist of effect amount.Compositions described herein or its pharmaceutically acceptable addition salt or hydrate are permissible According to method of disclosure use various route of administration or mode be delivered to curee with avoid or reduce unnecessary Side effect.Described pharmaceutical composition can be formulated into solid or liquid form Orally administered.

The well known by persons skilled in the art various compositions not disturbing the function of pharmaceutically acceptable carrier can be to have Effect amount is optionally included in pharmaceutical composition.Generally, lubricant, binding agent, gelatin and/or disintegrating agent are suitable 's.Other optional compositions include buffer, preservative, tension regulator, antioxidant, are used for regulating viscosity or serving as increasing The polymer of amount agent, and excipient, and like this.The those of ordinary skill of pharmaceutical field other conventional additive bags known Include but be not limited to, wetting agent, softening agent, stabilizer, and dyestuff, and if the treatment of these additives not interference medicament compositions Character just can be used.The method using pharmaceutical field generally known easily prepares described pharmaceutical composition.Can individually or With the compound of other disclosure combination and/or with other treatment or prevention IBD therapeutic combination use described compound or Its pharmaceutically acceptable salt and/or hydrate.

In other embodiments, disclosure theme provides the medicine box of the pharmaceutical composition including the disclosure.Medicine box The pharmaceutical composition of agonist or antagonist or its salt is comprised including at least one.Medicine box also includes for accommodating compositions Container.In some embodiments, medicine box also include curee is used at least one comprise agonist or antagonist or its The guide of the pharmaceutical composition of salt.

Suitably route of administration includes but not limited to, sucks, percutaneous, oral, rectum, through mucous membrane, intestinal, and parenteral is executed With, including intramuscular, subcutaneous, and intravenous injection.In treatment and/or diagnostic application, the compound of the disclosure can be formulated Become for various methods of application, use including (topical) with external of system or (1ocalized) of local.Technology and Formula generally can be at Remington:The Science and Practice of Pharmacy (20^th ed.) Lippincott, Williams & Wilkins (2000) find.

Compound according to the disclosure is effective in a wide dosage range.Such as, treat adult time, dosage from 0.01 arrives 1000mg, from 0.5 to 100mg, and the every day from 1 to 50mg.Definite dose-dependant is executed in route of administration, compound Form, curee to be treated, the body weight of curee to be treated, and the preference of the doctor in charge and experience.

Pharmaceutically acceptable salt is usually well known to those of ordinary skill in the art, and can include, such as but does not limits In, acetate, benzene sulfonate (benzenesulfonate), benzene sulfonate (besylate), benzoate, bicarbonate, wine Stone acid hydrogen salt, bromide, Ca-EDTA, d-camphorsulfonic acid salt, carbonate, citrate, edetate, Ethanedisulphonate (edisylate), Estolate (estolate), esilate (esylate), Rhizoma Corydalis Hydrochlorate, gluceptate (gluceptate), gluconate, glutamate, Glu, to hydroxyl acetylamino phenyl-arsonate (glycollylarsanilate), hexylresorcin salt (hexylresorcinate), Hai Baming salt, hydrobromate, hydrochloric acid Salt, hydroxynaphthoate (hydroxynaphthoate), iodide, isethionate (isethionate), lactate, lactobionic acid Salt, malate, maleate, mandelate, mesylate, mucate (mucate), naphthalene sulfonate (napsylate), nitre Hydrochlorate, pamoate (pamoate) (embonate (embonate)), pantothenate, phosphoric acid/diphosphate, polygalacturonic Hydrochlorate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, or 8-chlorine tea Alkali salt (teoclate).Other pharmaceutically acceptable salts can be at such as Remington:The Science and Practice of Pharmacy(20^thEd.) Lippincott, Williams & Wilkins (2000) find.Pharmaceutically Acceptable salt includes, such as, acetate, benzoate, bromide, carbonate, citrate, gluconate, hydrobromic acid Salt, hydrochlorate, maleate, mesylate, naphthalene sulfonate, pamoate (embonate), phosphate, salicylate, amber Amber hydrochlorate, sulfate, or tartrate.

According to treated concrete disease, these agent can be formulated into liquid or solid dosage form and by either systemically or locally Use.It is known to those skilled in the art that these agent can be delivered with such as timing or lasting sustained release forms.For preparation and The technology used can be at Remington:The Science and Practice of Pharmacy (20^th ed.) Lippincott, Williams & Wilkins (2000) find.Suitable approach can include being administered orally, oral cavity, suction-type spray Agent, Sublingual, rectum, percutaneous, vagina, through mucous membrane, nose or intestinal use；Potential delivery, including in intramuscular, subcutaneous, marrow In injection, and sheath, in direct ventricle, in intravenous, intraarticular, breastbone, intrasynovial, in liver, intralesional (intralesional), intracranial, intraperitoneal, intranasal or intraocular injection or other delivery modes.

For injection, the medicament of the disclosure can be formulated and be diluted in aqueous solution, the buffering of such as physiological compatible Liquid such as Hank ' s liquid, Ringer's mixture, or normal saline buffer solution.For mucosal administration, use in the formulation and be suitable for Needed for the penetrating agent of barrier that penetrates.These penetrating agent are generally known in the art.

Pharmaceutically acceptable inert carrier is used the compounds as disclosed herein being used for disclosure practice to be configured to Be suitable to the dosage (dosage) of systemic application in the scope of the present disclosure.Use the carrier of suitably selection and suitably manufacture reality Trampling, the compositions of the disclosure, particularly those preparations become the compositions of solution, can be in parenteral administration, such as by quiet Arteries and veins is injected.Can use and be easily configured to compound by pharmaceutically acceptable carrier well known in the art to be applicable to be administered orally Applied dose.These carriers make the compound of the disclosure can be formulated as sheet, ball, capsule, liquid, gel, syrup, serosity, Suspension and like this, so that curee to be treated (such as patient) is orally ingested.

For nose or inhalation delivery, the agent of the disclosure can also use method known to those skilled in the art to prepare, And be possibly including, but not limited to, such as dissolve, dilute or dispersed substance such as saline, preservative (such as benzylated polyol), absorption Accelerator and fluorocarbon.

The pharmaceutical composition being suitable for the disclosure includes following compositions: including the active component effectively measured to reach To its expection purpose.The determination of effective dose in the limit of power of those skilled in the art, in particular according to provided herein in detail Thin disclosure.

In addition to the active component, these pharmaceutical compositions can include help reactive compound be processed into pharmacy can The suitable pharmaceutically acceptable carrier of goods, including excipient and adjuvant.Being configured to Orally administered goods can be Tablet, sugar-coat agent, capsule or the form of solution.

The pharmaceutical preparation orally used can pass through mixed active compound and solid excipient, and optionally grinding obtains Mixture, and if it is required, after adding suitable adjuvant, process granulate mixture, obtain tablet or sugar-coat core.Close Suitable excipient, particularly, filler such as sugar, including lactose, sucrose, mannitol, or sorbitol；Cellulosics, such as beautiful Rice starch, wheaten starch, rice starch, potato starch, gelatin, Tragacanth, methylcellulose, hydroxypropyl methyl cellulose, carboxylic Sodium methide cellulose (CMC) and/or polyvinylpyrrolidone (PVP: polyvidone).It is possible if desired to interpolation disintegrating agent, example Polyvinylpyrrolidone, agar or alginic acid or its salt such as sodium alginate such as crosslinking.

Suitable coating is provided to sugar-coat core.For this purpose it is proposed, the sugar juice of concentration can be used, it can optionally include Arab Glue, Talcum, polyvinylpyrrolidone, 934 P gel (carbopol gel), Polyethylene Glycol (PEG) and/or titanium dioxide, Paint solution (1acquer solution), and suitably organic solvent or solvent mixture.Tablet or dragee coatings can be given Add dyestuff or pigment, to differentiate or to characterize the different combination of active compound doses.

The pharmaceutical preparation of orally available use includes the sucking fit capsule being made up of gelatin, and by gelatin and plasticizer The capsule of the soft sealing that (such as glycerol or sorbitol) is made.Sucking fit capsule can include and filler (such as lactose), viscous The active component that mixture (such as starch) and/or lubricant (such as Talcum or magnesium stearate) and optional stabilizer mix.At flexible glue In capsule, reactive compound can be dissolved in or be suspended in the poly-second of suitable liquid, such as fatty oil, liquid paraffin or liquid two Alcohol (PEG).It addition, stabilizer can be added.

According to the concrete disease to treat or to prevent or morbid state, generally it is applied the volume treating or preventing disease Outer therapeutic agent can be used together with the agonist of the disclosure and/or antagonist.Therefore, extra agent can be with the disclosure NAChR α 7 antagonist and/or nAChR α 7 agonist are incorporated in pharmaceutical composition.These extra agent can be as repeatedly giving A part for regimen (multiple dosage regimen) and the compositions separate administration comprising inhibitor.Alternatively, this A little agent can be a part for the one-pack type together with the inhibitor mixed in single compositions.

Although employ specific nomenclature at this, they only use and not for limit in the sense that describing common The purpose of system.Unless otherwise defined, all technology as used herein and scientific terminology have and the basis belonging to this theme described The identical implication that field those of ordinary skill is generally understood that.

Following the patent law tradition of long-time existence, term " (a) ", " one (an) " and " being somebody's turn to do " are when at the application bag Include and refer to " one or more " when claim uses.It is thus possible, for instance mention " curee " to include multiple curee, Unless context clearly points to (the most multiple curee) on the contrary, etc..

Running through this specification and claims, term " includes (comprise) ", " including (comprises) " and " bag Include (comprising) " use in the sense that non-excluded, unless the context requires otherwise.Similarly, term " comprise " and Grammatical variants is intended to not limit, and is not excluded for be substituted or add to cited item so that the item enumerated is enumerated Other similar items.

For the purpose of this description and in the appended claims, except as otherwise noted, all in this specification with power Profit claim in use expression amount, size, size, ratio, shape, formula, parameter, percentage ratio, parameter, quantity, feature and The numeral of other numerical value, it is thus understood that the most all modify with term " about ", even if term " about " is likely not to have Occur with numerical value, amount or scope clearly simultaneously.Therefore, unless made contrary explanation, in following description and appended power In profit claim, the digital parameters of statement is not also without being accurately, but can be on demand approximation and/or bigger or Smaller than desired, depend on seeking the character needed for being obtained by disclosure theme, it reflects tolerance, conversion factor, Round off, measurement error and like this, and other factors well known by persons skilled in the art.Such as, term " about ", when finger value In time, can be intended to include away from the following variation specifying value: in some embodiments for ± 100%, is in some embodiments ± 50%, it is ± 20% in some embodiments, is ± 10% in some embodiments, be in some embodiments ± 5%, it is ± 1% in some embodiments, is ± 0.5% in some embodiments, and be in some embodiments ± 0.1%, this type of variation is adapted for disclosed method or the compositions of the application disclosure.

And, when the scope with one or more numerals or numeral is used in combination, term " about " should be understood that Refer to all these numeral, including all in the range of numeral and by extend its boundary higher or lower than described numerical value and repair Change its scope.The enumerating of digital scope carried out by terminal includes all numerals, and such as all integer, is included into including it The mark of described scope, (such as, 1 to 5 enumerate includes 1,2,3,4 and 5, and its mark, such as 1.5,2.25,3.75,4.1, With like this) and any scope in the range of described.

Embodiment

Carry with the representative embodiment putting into practice disclosure theme to those of ordinary skill in the art including the following example For instructing.In view of present disclosure and the substantially technical merit of this area, those of skill will appreciate that, the following example only purport In example, and the change of many, modifications and changes can be employed the scope without deviating from disclosure theme.Following synthesis Describe and specific embodiment is only intended to for purposes of clarity, and be not necessarily to be construed as preparing the disclosure otherwise Any type of restriction of compound.

Embodiment 1

Gene expression atlas shows the different expression patterns in IBD

From 17 patients, obtain peripheral blood and colon biopsy, patient's mean age be 46.53 (95% confidence level, 39.65-53.41) and suffer from the IBD (9 CD and 8 UC) of record.This group also includes 17 ages and gender matched not Affected normal healthy controls.Separate neutrophil cell and peripheral blood lymphocytes (PBMC), and extract total serum IgE.Cy labelling CDNA and the microarray hybridization of Genome Scale, described microarray comprises 21, the probe of 329 genes.After data normalization, Before carrying out correlation analysis, in each sample about 9,500 genes are expressed as three (3) individual marks on background value Accurate poor.Hierarchical clustering shows the robustness of the analysis method for sample classification, its display, there is gene table in each subgroup Reach the different clusters (see Fig. 1) of collection of illustrative plates.Discrimination function analysis (DFA) is used to identify the expression in immunocyte at utmost Distinguish the gene of this group.Alex P., Szodoray P., Knowlton N., Dozmorov I.M., Turner M., Frank M.B., Arthur R.E., Willis L., Flinn D., Hynd R.F., Carson C, Kumar A., El- Gabalawy H.S., Centola M., " Multiplex serum cytokine monitoring as a prognostic Tool in rheumatoid arthritis, " Clin Exp Rheumatol 25:584-592 (2007).

In the gene of 96 differential expressions, strict DFA is used to select 10 genes as having supreme power for suffering from Classification between person and comparison differentiates.It is interesting that in the 3D figure of DFA root, UC and CD patient and comparison are grouped into 3 Different positions, shows that this is direct and full gene is expressed and changes in gene expression and the interior heterogeneity of group between the group of disease pathology Proportional (Fig. 2).NAChR α 7 is one in the gene of 10 differential expressions.

Embodiment 2

NAChR α 7 expression pattern of the difference identified in CD is compared to UC

Genetic microarray analysis shows, compared with the control, and the nAChR α 7 in the neutrophil cell and PBMC of CD patient Significantly raised, and the nAChR α 7 in the neutrophil cell of UC patient and PBMC has been lowered (Fig. 3, left figure).Use QRT-PCT confirms that (Fig. 3, right figure: CD change multiple compared to UC exists at the CD expression compared to the nAChR α 7 in UC It is statistically identical).Because nicotine is the main active component in smoking, and nAChR α 7 is only to compare at UC By the nAChR of differential expression in CD, it is different in smoker and non smoker that these data display nAChR α 7 is probably regulation The pro-inflammatory effect to CD and crucial participant to the antiinflammatory action in UC.

Embodiment 3

The different immunomodulating collection of illustrative plates of the nicotine produced by nAChR α 7 mediation in CD is compared to UC

In view of the nAChR α 7 differential expression (Fig. 3) that CD is unique compared in UC and healthy person, carry out preliminary research with Investigate the function contribution of nAChR α 7 and at CD compared to the immunomodulating collection of illustrative plates in UC.Although IBD is the most studied for T Cell-mediated disease, enteric microorganism of the same clan/B/T cell interaction is important for the pathogeny of disease.Research Immortalization (EBV-converts) bone-marrow-derived lymphocyte of UC, CD patient and unaffected comparison, to evaluate the immunity tune of nAChR α 7 Joint collection of illustrative plates.Cell is carried out nicotine (2 μMs and 20 μMs) and nAChR α 7 selective antagonist alpha-bungarotoxin (Sigma.St.Louis) stimulation and the suppression of (2nM and 20nM) is studied, and described cell is cultured and maintains 1 × 10⁶Individual carefully The final concentration of born of the same parents.(bioassay sandwich ELISA, it includes and for target egg protein array system based on sandwich immunoassay The microsphere of the dyeing of white monoclonal antibody specific coupling), as it was previously stated, Alex P., Szodoray P., Knowlton N., Dozmorov I.M., Turner M., Frank M.B., Arthur R.E., Willis L., Flinn D., Hynd R.F., Carson C, Kumar A., E1-Gabalawy H.S., Cento la M., " Multiplex serum cytokine Monitoring as a prognostic tool in rheumatoid arthritis, " Clin Exp Rheumatol 25:584-592 (2007)；Nakayama T., Hieshima K., Nagakubo D., Sato E., Nakayama M., Kawa K., Yoshie O., " Selective induction of Th2-attracting chemokines CCL17 and CCL22 in human B cells by latent membrane protein 1 of Epstein-Barr virus, " J Virol 78:1665-1674 (2004), is used for measuring the most multiple time point (24,48,72,96 and 120 hours) and releases Be put into culture medium 22 cells, cytotoxic, the cytokine of body fluid and the level (Fig. 4) of chemotactic factor.Use single Protein ELISA carries out the checking of multiple experiment.In these 22 variablees, the immune mediator (immune of the lymphocyte that EBV converts Mediator) the secretion notable change between unaffected comparison and IBD is only in 5 cytokine/chemotactic factors It is significantly (Fig. 4) (reproducing in two independent experiments).This collection of illustrative plates is report " low Th1, high Th2 chemotaxis " B lymph The typical case that the immunity of blast cell generates.Nakayama T., Hieshima K., Nagakubo D., Sato E., Nakayama M., Kawa K., Yoshie O., " Selective induction of Th2-attracting chemokines CCL17 and CCL22 in human B cells by latent membrane protein 1 of Epstein-Barr Virus, " J Virol 78:1665-1674 (2004)；Burdin N., Peronne C, Banchereau J., Rousset F., " Epstein-Barr virus transformation induces B lymphocytes to produce human Interleukin 10, " J Exp Med 177:295-304 (1993)；Vockerodt M., Pinkert D., Smola- Hess S., Michels A., RansohoffR.M., Tesch H., Kube D., " The Epstein-Barr virus oncoprotein latent membrane protein 1 induces expression of the chemokine IP- 10:importance of mRNA half-life regulation, " Int J Cancer 114:598-605 (2005).

Use the nicotine pretreatment 120 hours display of various dose, at CD compared in UC, proinflammatory cytokine/chemotactic Factor IL-8 and the significant generation of MIP-1, and at CD compared in CD, what anti-inflammatory cytokines IL-10 generated significantly presses down System (Fig. 4 A), this shows that nicotine has different proinflammatory and chemotactic effects in CD.Nicotine suppresses the effect (figure of IL-10 in CD 4A) also demonstrating a potential mechanism, it also develops in the mice that gene target IL-10 knocks out be extensively examined Chronic CD sample ileocolitis consistent, and with the curative effect in the IL-10 animal model to some colitis of report mutually Cause.Kuhn R., Lohler J., Rennick D., Rajewsky K., Muller W., " Interleukin-10- Deficient mice develop chronic enterocolitis 2, " Cell 75:263-274 (1993)；Ribbons K.A., Thompson J.FL, Liu X., Pennline K., Clark D.A., Miller M.J., " Anti- inflammatory properties of interleukin-10 administration in hapten-induced Colitis, " Eur J Pharmacol 323:245-254 (1997).Owing in CD, alpha-bungarotoxin is with 100% significantly Reversing the effect of nicotine, these data also show the pro-inflammatory effects of the mechanism in CD by nAChR α 7 mediation.In fresh separated Lymphocyte and the similar experiment well afoot studying the cytokine profiles that nAChR α 7 mediates in neutrophil cell. Above-mentioned discovery shows the nAChR α 7 key effect in terms of regulation nicotine effect strongly, and shows that nAChR α 7 is as CD The notable potentiality of the therapeutic target for the treatment of.

Use nicotine or " UC sample " mouse colitis (disease of nAChR α 7 specific agonist pretreatment prevention DSS induction Phenotype is similar with mankind UC) development, and significantly deteriorate colitis (disease phenotype and mankind's CD class of " CD sample " TNBS induction Like).

For obtaining the nAChR α 7 direct card as the main moderator coordinating the nicotine opposite effect in CD and UC process According to；The mice IBD model (DSS and TNBS colitis) using well-established chemical induction carried out series of experiments (Fig. 5- 13), described mice IBD model is widely used in the preclinical study of IBD curative drug.For CD sample mouse model, use BALB/c mouse induction DSS colitis.For CD sample mouse model, use BALB/c mouse induction TNBS colitis.C57/B6 Or BALB/c mouse, 6-8 week old, use nicotine (nAChR α 7 non-specific ligand；6mg/kg) or nAChR α 7 specific agonism Agent (PNU；8.3mg/kg) pretreatment (peritoneal injection every day) week.Comparison is mice (each process not having pretreatment Organize and a total of be more than 10 mices).Then, use TNBS (600 μ g) inducing mouse development CD sample colitis, or use DSS (2.5%) inducing mouse development UC sample colitis.As it is shown in figure 5, for the disease activity index of acute colitis model (DAI), nicotine or 5 days severe exacerbation CD sample TNBS colitis of PNU pretreatment.Completely contradict, but nicotine and PNU prevent DSS The UC sample colitis of induction, wherein PNU is the most more effective.Morphology and the histology of colon further demonstrate that DAI's Change (data do not show).The preclinical study result accurate simulation of these mouse models is clinically by the smoking of well-characterized CD is harmful to but useful in UC by the opposite effect to mankind IBD, i.e. smoking.These data are established: the morbidity to CD of (1) nicotine The effect of mechanism and the therapeutic benefit to UC；(2) nAChR α 7 is anti-as the difference of main on-off control nicotine (smoking) Should, or be proinflammatory (as in CD) or be antiinflammatory (as in UC).Agonist GTS-21 has similar prevention DSS induction The effect of development of CD sample colitis.

Embodiment 4

NAChR α 7 agonist is effective at the treatment camber of chronic " UC sample " DSS colitis

NAChR α 7 agonist (PNU and GTS-21) not only has the ability to prevent the knot of DSS induction in acute colitis model Enteritis (Fig. 5), they can also in chronic colitis model effective reverse disease process (Fig. 6), described chronic colitis mould Type is considered more similar with mankind UC.The legend of Fig. 6 describes the induction of chronic UC sample colitis.As shown in Figure 6, it is shown that The PNU impact on the DAI of acute and chronic colitis model.PNU process substantially eliminates the inflammation in two kinds of models.

Embodiment 5

NAChR α 7 antagonist MLA is effective to prevention CD sample TNBS colitis height

The observed result lowered in UC based on nAChR α 7 and raise in CD, it is undesirable to by any specific Theoretical constraint, it is assumed that nAChR α 7 specific antagonists can make the functionally inactive of nAChR α 7, and therefore CD is had therapeutic effect. Therefore, BALB/c mouse is used to have detected the nAChR α 7 specific antagonists MLA effect to CD sample TNBS colitis.Really, as Shown in Fig. 7, find mice uses nAChR α 7 specific antagonists MLA pretreatment within 5 days, can effectively prevent mice to develop TNBS The colitis (Fig. 7) of induction.Importantly, nAChR α 7 agonist PNU processes not only development to colitis does not has effect, and Deteriorating condition.The agonist nicotine more weak to nAChR α 7 specificity is effective to prevention UC sample DSS colitis, but develops disease There is no effect (Fig. 5) yet.These results simulate smoking effect in UC and CD.Merge the data shown in Fig. 5 and Fig. 6, this A little results display nAChR α 7 specific antagonists (MLA) have an effective therapeutic benefit to CD, but nAChR α 7 specific agonism Agent (PNU and GTS) has effective therapeutic effect to UC.Also demonstrate MLA and CD sample chronic TNBS colitis (is used BALB/c Mice) high therapeutic effect (data do not show), its effect and PNU (figure similar to the effect of UC sample DSS colitis 6)。

Embodiment 6

NAChR α 7 antagonist is the most effective to the treatment of chronic " CD sample " TNBS colitis

NAChR α 7 antagonist (MLA) not only has the ability to prevent acute TNBS colitis (Fig. 7), and they can also be the most inverse Turning the disease process (Fig. 8) of chronic TNBS colitis model, wherein chronic TNBS colitis model is considered and mankind CD more phase Seemingly.The induction of chronic CD sample colitis (Alex et al., 2009 IBD, 15 (3), 341-352) as previously mentioned.It is right that agonist processes Disease activity does not affect, but antagonist processes and substantially eliminates the inflammation in two kinds of models.Mice seems the most also It is undistinguishable with the control mice of Ethanol Treatment.The histologic analysis (Fig. 9) using H&E dyeing to carry out show further The nAChR α 7 specific antagonists highly effective therapeutic effect to CD sample colitis.Biochemical method can also be passed through Find out therapeutic effect, because DSS colitis being used agonist or using antagonist to process TNBS colitis, acutely change Cytokine profiles is from morbid state to normal healthy state (Figure 13).

Embodiment 7

Agonist PNU and antagonist MLA does not has therapeutic to imitate in nAChR α 7 knock-out mice being induced generation colitis Really, it is specific for further illustrating described medicine to nAChR α 7 receptor.

Although the data coming from the disclosure of cell model and mouse colitis model understand identifies nAChR α 7 conduct CD is in response to pathogenetic key point of nicotine, but the probability that other nAChR are not included can not be got rid of completely, because of The specificity of nAChR α 7 agonist/antagonist for using may be not limited to only nAChR α 7.In order to get rid of this probability, Test nAChR α 7 agonist and antagonist in nAChR α 7 knock-out mice (nAChR α 7-KO), in described knock-out mice, if Described dose is only specific to nAChR α 7, then the development of chemical induction colitis will not be affected by these agent.Find, The colitis more sensitive (Figure 10) that DSS or TNBS is induced by nAChR α 7-KO mice.The more important thing is, agonist (PNU) or Therapeutic agent (the figure of the colitis that antagonist (MLA) all can not be induced as effective DSS or TNBS in nAChR α 7-KO mice 10).These data show, the target of these medicines is specific to nAChR α 7.

Embodiment 8

The antiphlogistic effects of DSS colitis is completely eliminated by nAChR α 7 agonist by vagotomy, it is provided that at IBD Middle vagus nerve plays first deterministic internal evidence of pivotal role in the anti-inflammatory pathway that nAChR α 7 mediates

Vagal outgoing signal can be controlled by brain network, suppresses cytokine by cholinergic anti-inflammatory pathway Generation, wherein cholinergic anti-inflammatory pathway depends on the nAChR α 7 on non-neuronal cells particularly macrophage.Led to by this Road, vagal stimulation prevention detrimental effect of release of cytokines in multiple inflammatory diseases, inflammatory diseases includes deteriorated blood Disease, endotoxemia, ischemia, hemorrhagic shock, arthritis and postoperative enteremphraxis.Although having shown that experienced by vagus nerve cuts The colitis induced DSS and TNBS except the mice of art is more sensitive, but directly not experimental evidence by vagus nerve and The regulation of IBD is specifically connected by nAChR α 7.Data owing to being shown before clearly show that UC is resisted by nAChR α 7 Pathogenetic specificity of scorching effect and CD participates in, and further determines that whether the specific effect of this nAChR α 7 depends on Vagus nerve.

The left vagus nerve of the gastroesophageal junction of the subdiaphragmatic vagus nerve of the C57B/6 mice (7 week old) under anesthesia is divided Prop up (about 5-10mm) and carry out vagotomy (Figure 11 A).Experienced by vagotomy mice 7 days time Between recover, then use nAChR α 7 agonist with IP, continue 5 days, carry out DSS process (7 days) afterwards with inducing colitis.As Shown in Figure 11 B, the mice that experienced by vagotomy has completely lost nAChR α 7 agonist protection work to colitis With.These data show, the antiphlogistic effects to IBD of nAChR α 7 mediation places one's entire reliance upon and has the vagus nerve of function.

Embodiment 9

Agonist PNU is to treatment GPX 1/2 The colitis height of DKO mediation is effective

Owing to the major part of experiment in vivo before is carried out in experimental colitis model, examine nAChR α 7 The colitis that treatment is mediated by agonist by genetic flaw is effectively assumed.Two glutathion peroxidase (GPX) Gpx1 With the mice of Gpx2 defect [Gpx1/2-is double knocks out (DKO) mice] at C57BL/6 and 129S1/SvJ (B6.129) base mixed Because of development ileocolitis spontaneous under background.Lee D.H., Esworthy R.S., Chu C, Pfeifer G.P., Chu F.F., “Mutation accumulation in the intestine and colon of mice deficient in two Intracellular glutathione peroxidases, " Cancer Res.66 (20): 9845-51 (October 15 in 2006 Day).The subgroup of B6.129 Gpx1/2-DKO mice is to developing ieoocolon inflammation 6-8 week, and develops ieoocolon by 6-9 month Tumor.But comparison (coming from C57BL/6 and 129S1/SvJ (B6.129) genetic background of mixing) does not develop the body of colitis Levy, find that the development of GPX1/2DKO mice has Clinical Activity scoring and is about the serious colitis (Figure 12) of 9.Once tie Enteritis sign is obvious immediately, uses nAChR α 7 agonist and nAChR α 7 antagonist in GPX1/2DKO mice.Using nAChR α In the GPX1/2DKO mice that 7 agonist process, the development of disease (colitis) significantly reduces, and shows nAChR α 7 agonist PNU The colitis height mediated treatment by GPX1/2DKO is effectively (Figure 12).But use nAChR α 7 antagonist or saline treatment The significance change that GPX1/2DKO mice is not shown in the scoring of colitis.These data show that nAChR α 7 agonist is to knot The chemistry of enteritis and the treatment of genetic model are the most effective.

Embodiment 10

PNU does not has observable side effect to major organs and the behavior of mice.

Under the dosage that preclinical study uses, mice displaying is had no side effect by PNU medicine.All major organs, including Heart, kidney, liver, small intestinal and large intestine, spleen, outward appearance all seems normal.Force swimming test: mouse is forced at cylinder Swim 6 minutes, such as Porsolt R.D., Bertin A., Jalfre M., " Behavioral despair in describing device Mice:a primary screening test for antidepressants, " Arch Int Pharmacodyn Ther Described by 229:327-336 (1977), to evaluate whether medicine changes the behavior (movable as reading) of mice. ZomkowskiA.D., Santos A.R., Rodrigues A.L., " Evidence for the involvement of the opioid system in the agmatine antidepressant-like effect in the forced Swimming test, " Neurosci Lett 381:279-283 (2005).As shown in figure 14, significance is not observed No matter mice Behavioral change, pass through or without drug treating.The mice of only DSS inducing colitis shows the motionless of significance. Use the process prevention colitis of agonist PNU, and reverse the motionless of DSS mice the most completely.This result shows, NAChR α 7 agonist does not have side effect to the behavior of the mice of research.Other researchs of these nAChR α 7 specific drugs, Including toxicity and pharmacokinetics, studying in carrying out.

In sum, the compound of the disclosure provides single therapeutic target, nAChR α 7:nAChR α 7 agonist to CD and UC (such as PNU and GTS-21) is curative drug highly effective to UC, and CD is had by nAChR α 7 antagonist (such as MLA) The treatment potentiality of significance.

List of references

All publications, patent application, patent and other lists of references mentioned in this manual are to disclosure master The instruction of the level of the technical staff in the field belonging to topic.All publications, patent application, patent and other lists of references are equal To be incorporated herein by reference, with each single publication, patent application, patent and other lists of references by specifically and single As solely pointing out to quote and being incorporated to.Although being appreciated that a large amount of patent application mentioned herein, patent and other lists of references, But this quoting does not constitutes the recognizing an of part that any one in these files is formed the common general knowledge in this area.

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Although for clearness of understanding by illustrating that the method with example describes aforesaid theme with some details, It will be understood by those skilled in the art that some changes and amendment can be put into practice within the scope of the appended claims.

Claims (3)

1. methyllycaconitine (MLA) is used for treating Crohn disease (CD) in the curee having corresponding treatment to need in preparation Purposes in medicine, when described medicine is applied, in described MLA at least one cell with the effectively described curee of regulation The amount of activity of nAChR α 7 treat the CD in described curee.

Purposes the most according to claim 1, wherein said treatment includes preventing CD development in described curee.

3. methyllycaconitine (MLA) develops colorectal cancer in preparation for reducing in the curee with Crohn disease (CD) Risk medicine in purposes, when described medicine is applied, methyllycaconitine (MLA) is effectively to regulate one or more stomaches The inflammation that the amount of the activity of the nAChR α 7 in intestinal immune cell changes in one or more gastrointestinal tract immunocytes described is anti- Should, thus reduce the risk of development colorectal cancer.