CN102743365A - Use of compound or its pharmaceutically acceptable salt in prevention and treatment on senile dementia - Google Patents

Abstract

The invention discloses a use of a compound or its pharmaceutically acceptable salt in prevention and treatment on senile dementia. The compound is N'-{5-[acetyl(hydroxyl)amido]pentyl}-N-[5-({4-[(5-aminopentyl)( hydroxyl)amido]-4-keto-butyryl}amido)pentyl]-N-hydroxysuccinic acid amide. The compound or its pharmaceutically acceptable salt can be used as a drug for preventing and treating senile dementia. The compound or its pharmaceutically acceptable salt can penetrate a blood brain barrier and then bind with a heavy metal to form a chelate thereby inhibiting production of amyloid in the brain, preventing polymerization and accumulation of amyloid, dissolving produced amyloid plaques and promoting amyloid degradation.

Description

A kind of chemical compound or its pharmaceutically acceptable salt are in the application of control senile dementia

Technical field

The present invention relates to a kind of new purposes of medicine, more particularly, relate to the application of acceptable salt on the control senile dementia on a kind of chemical compound or its materia medica

Background technology

Alzheimer disease is main popular a kind of chronic neurodegenerative disease in the old people.Wherein modal be Alzheimer's disease (Alzheimer ' s disease, AD).By sick scholar of Germanism and neuropathologist Alois Alzheimer this can not the treatment described first in 1906; Retrograde human terminal point disease (Berchtold NC, Cotman CW.Evolution in the conceptualization of dementia and Alzheimer ' s disease:Greco-Roman period to the 1960s.Neurobiol.Aging 1998; 19 (3): 173-189.).In most cases, AD falls ill in the old people of over-65s, the early onset AD that the accidental middle age takes place.2006, it is ill that AD has involved the whole world 2,660 ten thousand people.Expecting per 85 philtrums in the year two thousand fifty whole world will have 1 AD patient, and the whole world number of getting involved surpasses 100,000,000, and (10,588 ten thousand people see Brookmeyer, R; Johnson, E; Ziegler-Graham, K; Arrighi, HM.2006 prevalence estimate:Forecasting the global burden of Alzheimer ' s disease.Alzheimer ' s and Dementia.2007; 3 (3): 186-91.).

AD is that to lose with brain cell be a kind of progressivity disease of characteristic, causes serious dull-witted and dead.Though the course of disease of everyone AD all has difference, AD patient has many common symptoms.It is relevant with the age worried and nervous that observed the earliest symptom is often takeed for; The modal symptom of commitment is study, memory and cognitive disorder; Along with the progress of the state of an illness, the symptom of generation comprises confusion of thinking, irritability and aggressivity, and anxious state of mind, language breaks, the longterm memory forfeiture; The body function forfeiture takes place late period gradually, finally causes death.When suspection suffers from AD; Normally according to the behavior assessment, recognition tests, if possible; Confirm diagnosis (Waldemar G through brain scans; Dubois B, Emre M, et al.Recommendations for the diagnosis and management of Alzheimer ' s disease and other disorders associated with dementia:EFNS guideline.Eur J Neurol.2007; 14 (1): e1-26.).AD patient individual's prognosis is difficult to assessment, and according to statistics, AD patient's average life expectancy is about 7 years, only lives through 14 years less than 3 percent people.

Present stage, the pathogenesis of AD disease is still unclear.Research shows; Relevant with the AD progress is that amyloid plaques and fibre matting (Tiraboschi P are arranged in the brain; Hansen LA; Thal LJ, Corey-BloomJ.The importance of neuritic plaques and tangles to the development and evolution of AD.Neurology 2004; 62 (11): 1984-1989.).The therapeutic scheme of using at present only has little effect, does not still have the medicine that can postpone or stop disease progression.By 2008; The clinical trial of more than 500 treatment AD possibility drug identification has been arranged; Is but still do not have any testing drug or Intervention Strategy (the http://www.clinicaltrials.gov/ct2/results that clear and definite curative effect is arranged? Term=alzheimer, " Alzheimer ' s Disease Clinical Trials " .US National Institutes of Health.2008.).The symptomatic treatment that adopts at present, its curative effect still has a question.And the medicine of current any clinical trial of carrying out can not produce breakthrough symptomatic treatment in the recent period.If there is not efficacious therapy, patient's medical expense, our the existing medical budget of might crushing.Therefore, unremitting research and development efficacious therapy medicine still has the important clinical meaning at present.

Known amyloid, promptly A β (amyloid-β, A β) peptide is piled up, and in the AD pathogenic course, has brought into play important effect.Although the target of the experiment medicine of research and development is that A β produces or remove the A β in the brain in the inhibition brain usually.But discover that the abnormal deposition of copper, ferrum, zinc is relevant with the A β involved area in the AD brain.

The clinical diagnosis of alzheimer disease is to find that amyloid is deposited in appearance and accumulation in the brain.Amyloid mainly is found in neuronic stub area, is heterogeneous deposition in the morphology, is also referred to as senile plaque.The formation of senile plaque is relevant with the performance of the sings and symptoms of disease, comprises AD.After senile plaque forms, in neuronal cell, produce neurofibrillary tangles, the formation of neurofibrillary tangles is relevant with the deterioration of other symptoms of AD and dementia.

Report is pointed out; High concentration ferrum is regulated amyloid protein precursor (APP) conversion enzyme through ferritin; Changed the amyloid that the processing of α secretase produces; The deposition (Silvestri L, Camaschella C.A potential pathogenetic role of iron in Alzheimer ' the s disease.J Cell Mol Med.2008 that have caused A β; 12 (5A): 1548-1550.).The critical event of the early stage europathology of AD is a brain iron ion dysequilibrium, comprises oxidative stress, inflammatory process, amyloid deposition, pottery protein phosphorylation and neuronal cell periodic adjustment obstacle and apoptosis.The evidence that is increasing shows, the excessive accumulation of ferrum takes place in brain many neurodegenerative diseases, and the active iron of high concentration can increase oxidative stress, induces neuronal damage, increases endogenous and the toxicity external source poisonous substance.In fact, ferrum is piled up and oxidative stress is the early stage incident of AD, and this incident more approaches to occur the AD pathological process of significance and significant pathological changes.There is report to think that having the genotypic individuality of transferrins C1C1 has higher total iron binding capacity (TIBC); This has explained that the TIBC level increases; With iron concentration in its brain increase relevant, and in the brain high concentration iron ion more be prone to suffer from AD (Tang Lei, Yang Ze etc. Chinese neuroimmunology and neurological's magazine; 2005,12 (5): 321-322).

Therefore, reduce or eliminate the symptom that the intravital amyloid deposition of alzheimer disease patient possibly alleviate alzheimer disease.Present stage, carried out multiple trial and prepared the sedimentary medicine of removing amyloid.For example, WO 93/10459 discloses through giving the method for zinc bonding agent treatment alzheimer disease.The preferred compound of wherein mentioning is a phytic acid, sodium citrate, EDTA, 1,2-diethyl-3-hydroxyl. pyridine-4-ketone and 1-ethoxy-3-hydroxy-2-methyl-pyridine-4-ketone.DE 3932338 discloses chelating agen, like oxine, and the application in the treatment of alzheimer disease.US 5373021 discloses disulfiram and salt and analog, and disclosed chemical compound can be used for reducing the neurological damage that alzheimer disease especially causes.Yet, above-mentioned these chemical compounds all more or less have some shortcomings, for example, majority can not penetrate blood brain barrier in these chemical compounds, therefore almost can not arrive the zone of amyloid beta deposition.Though the penetrable blood brain barrier of disulfiram, shortcoming are them also is pure blockage.The most deep chelating agen of research is EDTA.Yet EDTA can not penetrate blood brain barrier equally, and thinks that it has sizable toxicity.In the AD pathogenesis, the amyloid pathological changes takes place, metal ion plays pivotal role in the redox reaction.Under the AD pathogenesis drives, promoted the metal-chelate mixture that research and development are useful on the AD treatment with good prospect.

Summary of the invention

The present invention is exactly to the problems referred to above, and the application on the control senile dementia of a kind of chemical compound or its pharmaceutically acceptable salt is provided.

In order to realize above-mentioned purpose of the present invention, the present invention adopts following technical scheme,

The structural formula of chemical compound is suc as formula shown in the I,

Formula I

Molecular formula is C ₂₅H ₄₈N ₆O ₈Molecular weight is 560.684; The chinesization formal name used at school is called N '-{ 5-[acetyl group (hydroxyl) amine] pentyl }-N-[5-({ 4-[(5-aminopentane base) (hydroxyl) amine]-4-ketone-bytyry } amine) pentyl]-N-hydroxy succinic acid amide, and English chemical name is: N '-[5-[acetyl (hydroxy) amino] pentyl]-N-[5-[[4-[5-aminopentyl (hydroxy) amino]-4-keto-butanoyl] amino] pentyl]-N-hydroxy-succinamide.

Described chemical compound or its pharmaceutically acceptable salt can be used as medicine and are used to prevent and treat senile dementia.

Described salt is mesylate.

Said chemical compound or its pharmaceutically acceptable salt see through blood brain barrier and heavy metal is combined into chelate, suppress amyloid generation in the brain, and blocking-up amyloid polymerization and accumulation are dissolved established amyloid speckle and promoted the amyloid degraded.

Described heavy metal is more than one in ferrum, copper, aluminum, the zinc.

During medication, can in said chemical compound or its pharmaceutically acceptable salt, add acceptable carrier or solvent on the materia medica.

Described solvent is that normal saline, distilled water or mass concentration are the 5-10% glucose.

Described carrier is more than one in starch, sucrose, mannitol, silicon derivative, carboxymethyl cellulose, gelatin, polyvinylpyrrolidone, glycerol, carboxymethyl starch sodium, crospovidone, low-substituted hydroxypropyl cellulose, paraffin, quaternary ammonium compound, hexadecanol, glyceryl monostearate, Kaolin, bentonite, calcium stearate, magnesium stearate, micropowder silica gel, the Pulvis Talci.

The using dosage of said medicine is the 2-80mg/kg body weight/day.

Beneficial effect of the present invention

The used medicine of the present invention is a marketed drugs, and safety is secure, in the test, zest and any untoward reaction do not occur.

The used medicine of the present invention has the ability that penetrates blood brain barrier, can be effectively be combined into chelate preventing amyloid aggregation with heavy metal, and can dissolves the sedimentary effectiveness of amyloid again.Result of the test shows: AD rat drug treatment group is compared with placebo group (having only normal saline), significantly reduces cortex behind the brain, the level of A β 1-40 in striatum and the Hippocampus; After 8 weeks of medication, proved further that through 16 all water maze tests it can significantly improve memory.Usefulness with AD treatment can reduce A β generation and/or promote A β to remove, and improves memory ability.

This medicine combines with the ferrum high selectivity; The neuroprotective that significance is arranged; The necrosis that prevents that six oxidopamine from causing in rat brain is arranged simultaneously, effectively prevent and the usefulness of the necrosis that the tyrosine hydroxylase positive neuron that recovers mice is caused by proteasome inhibitor breast Bao Su.Can reduce the active inhibition of ubiquitin protein enzyme body in significance ground.

Description of drawings

Fig. 1. medicine stimulates the APP of people SH-SY5Y cell and the electrophoretogram that tubulin is expressed influence to DMSO.

Fig. 2. medicine stimulates the APP of people SH-SY5Y cell and the cell survival rate figure that tubulin is expressed influence to DMSO.

Fig. 3 .AD animal model morris water maze laboratory is sought the platform trajectory diagram; 1) matched group; 2) small dose group; 3) heavy dose of group.

Fig. 4 .AD animal model medication front and back brain tissue slice figure is x200 a), the cerebral tissue pathological section figure behind the AD animal-use drug; B) the cerebral tissue pathological section figure before the x200, AD animal-use drug.

The specific embodiment

N '-{ 5-[acetyl group (hydroxyl) amine] pentyl }-N-[5-({ 4-[(5-aminopentane base) (hydroxyl) amine]-4-ketone-bytyry } amine) pentyl]-N-hydroxy succinic acid amide; English: N '-[5-[acetyl (hydroxy) amino] pentyl]-N-[5-[[4-[5-aminopentyl (hydroxy) amino]-4-keto-butanoyl] amino] pentyl]-N-hydroxy-succinamide, its structural formula does

This molecule has the ferric ion affinity, can be used as iron chelator.It can permeate through cell and blood-brain barrier: can avoid combining with the ferrous iron of hemoglobin, also can avoid causing the loss of whole body metal ion, comprise and avoid suppressing the key enzyme that brain relies on metal ion.In clinical, be used to treat the medicine of the too high disease of iron load.

The influence that embodiment 1 medicine is expressed A beta-amyloyd precursor protein (APP)

1, experiment material

Cell: people SH-SY5Y neuroblastoma cell, grind (Shanghai) available from giving birth to.

Medicine: N '-{ 5-[acetyl group (hydroxyl) amine] pentyl }-N-[5-({ 4-[(5-aminopentane base) (hydroxyl) amine]-4-ketone-bytyry } amine) pentyl]-N-hydroxy succinic acid amide is available from NOVARTIS company.

Pharmaceutical agent: DMEM culture medium (Gibco), dimethyl sulfoxide (DMSO), rabbit APP polyclonal antibody (A8717, Sigma).

Reference protein: 'beta '-tubulin monoclonal antibody (T4026, Sigma company), LDH test kit (Luo Shi), the M-PERe mammalian proteins extracts reagent (Pierce), the mixed liquor of protease inhibitor (Luo Shi).Two anti-and chemical luminescence reagent kits (Pierce company)

2, test method

(1) cell culture: people SH-SY5Y cell is added with final concentration in the DMEM culture fluid be that 10wt% hyclone and final concentration are 1wt% twin antibiotic adherent growth; Went down to posterity once in 4-7 days; Abandon culture fluid in the former bottle when going down to posterity, the trypsinization that adds final concentration and be 0.25wt% adds new culture fluid after 2 minutes.

(2) administration: when cell covers with the 40-50% culture bottle, be divided into 4 groups at random: it is that this medicine of 1wt%DMSO+ variable concentrations (is respectively 0 that negative control, experimental group add final concentration respectively; 1,5,10 μ M) processing; In identical cell culture condition (95%O2,5%CO2, humidity 85%; 37 ℃) under, incubated cell is 48 hours continuously.

(3) carry out SDS-PAGE and analyze, supernatant total protein concentration 1ug/ul, last appearance 20ul/ hole.The immunoblotting electricity goes to pvdf membrane behind the electrophoresis, presses test kit method and antibody hybridization colour developing, exposure imaging.Reference protein ('beta '-tubulin) has also carried out same detection, to guarantee the targeting specific influence of this medicine to APP, the visible Fig. 1 of result.

3, experimental result

(1) can know by Fig. 1, though, through this drug treating with without each group of the reference protein ('beta '-tubulin) of this drug treating between do not have significant different; But; Comparing A β 1-40 protein expression through this medicine (5,10 μ M) processed group and preceding two groups (0,1 μ M) group obviously reduces.Wherein handle two groups and compare indifference between the A β 1-40 protein expression through this medicine (0,1 μ M).

Only add 1%DMSO in the reference protein, do not add medicine.APP is an amyloid precursor protein.

(2) for guaranteeing that this medicine is not due to the cell death that causes because of this drug treating to the potential impact of APP protein expression, behind this drug treating cell, has detected cell survival rate with mtt assay.

The MTT colorimetry is a kind of method that detects cell survival and growth.It detects principle is that succinate dehydrogenase in the living cells mitochondrion can make exogenous MTT be reduced to water-insoluble bluish violet crystallization first a ceremonial jade-ladle, used in libation (Formazan) and is deposited in the cell, and dead cell does not have this function.First a ceremonial jade-ladle, used in libation in dimethyl sulfoxide (DMSO) the ability dissolved cell is measured its absorbance value with enzyme-linked immunosorbent assay instrument in the 490nm wavelength, can reflect living cells quantity indirectly.In certain cell number scope, the amount that the MTT crystallization forms is directly proportional with cell number.This method has been widely used in the activity detection of some bioactie agents, large-scale screening anti-tumor medicine, cell toxicity test and tumor radiosensitivity mensuration etc.Its feature is highly sensitive, economical.

With MTT kit measurement cell viability, the visible Fig. 2 of result.Can know by Fig. 2, under 3 kinds of various dose concentration, cell survival rate all be higher than 85%. this show that large and small dosage (5,10 μ M) all can reduce the A β 1-40 albumen in the neurocyte.

Embodiment 2

1, experiment material

(1) laboratory animal: 30 6 male SAMP8 mices of monthly age (Ministry of Public Health Gerontological Research Center institute), body weight 34.4 ± 2.3g.SAMP8 Mus average life is 299 days, 4～6 monthly ages reach ripe after date occur obviously old and feeble, be research aging and the good animal model of diseases associated with senescence (Morris R.Neurosci Meth, 1984,11:47-49.).

(2) main agents and instrument: medicine: N '-{ 5-[acetyl group (hydroxyl) amine] pentyl }-N-[5-({ 4-[(5-aminopentane base) (hydroxyl) amine]-4-ketone-bytyry } amine) pentyl]-N-hydroxy succinic acid amide, available from NOVARTIS company; SLY-WMSMorris water maze analytical system.A β 1-40 monoclonal antibody (DAKO), II, III antibody and DAB (available from doctor's moral company).

2, experimental technique

(1) administration: subcutaneous slow injection 1ml medicine, once a day, continuous 8 weeks; Laboratory animal is divided into 3 groups at random: (this is the amount of every day to dosage 75mg/kg/ daily weight for matched group injecting normal saline, medication therapy groups (heavy dose of group); During use, be to be dissolved in normal saline with medicine), this medication therapy groups (small dose group) dosage 25mg/kg body weight.(x ± s) expression adopts SPSS15.0 software to carry out statistical procedures to data, relatively uses variance analysis and rank test between group with mean ± standard deviation.

(2) Morris water maze test: after each is organized mice and treated for 8 weeks, carried out continuous 5 days (d) subsequent experimental in promptly the 57th day.

A. orientation navigation test: 1-4 days, from the place of entry of I, II, III, an IV4 quadrant, towards the pool wall entry, picked up counting the mice face simultaneously every day respectively, and mice is found platform and climbs up the required time of platform is escape latency.In official hour (90s), can not find platform like mice, then guide it into platform by the experimenter, behind the rest 30s, repeated trials.

B. space exploration test: the 5th day, remove platform, with mice in the I quadrant, face the pool wall entry, mice is passed through the number of times of original platform position in the opening entry 90s simultaneously.

When (3) experiment finishes, win the Mus brain, through the formalin fixed of mass concentration 10%, FFPE after the section, adopts the SABC method, detects the positive staining intensity of A β 1-40 (dull-witted albumen).

3, experimental result

Small dose group respectively has 1 dead mouse with heavy dose of group.

The Morris water maze test is to be widely used in the test method that detects the memory of animal space learning at present.What each organized the result relatively sees table 1.

Table 1 is respectively organized mice Morris water maze test result's comparison (x ± s)

Annotate: compare * P＜0.01 with matched group; Compare #P＜0.05 with the heavy dose group

Can know by table 1, compare that small dose group and heavy dose of group mice are found and obviously shorten (being P＜0.01) incubation period of platform with matched group; Wherein heavy dose of group mice is found and is significantly shorter than matched group and small dose group (all P＜0.05) incubation period of platform.The directed learning ability that shows heavy dose of group, small dose group mice all obviously improves than control group mice, and the directed learning ability of heavy dose of group mice improves particularly evident;

The event trace of small dose group and heavy dose of group mice is more prone to rest on the quadrant that original platform belongs to, and mostly the event trace of control group mice is round peripheral pool still.Can know by Fig. 3, compare that the number of times that small dose group (2) and heavy dose of group (3) mice are passed through the original platform position obviously increases (P＜0.05～0.01) with matched group (1).Show that small dose group and the spatial memory ability of heavy dose of group mice all are improved, more obvious with the action effect of heavy dose group especially.

The pathologic finding of cerebral tissue A β 1-40 protein content adopts commercially available A β SABC test kit, is undertaken by conventional method.As shown in Figure 4, the little dark brown A of 25mg/kg dose groups murine brain β 1-40 positive cell is expressed, and matched group greatly then has a large amount of dark brown A β 1-40 positive cells to express, heavy dose of group murine brain showed increased.

Can know that by above-mentioned experiment the medicine among the present invention gives more than the animal model 25mg/kg body weight, can reduce the dull-witted protein expression of A β 1-40 in the murine brain, help dementia.Morris water maze laboratory, animal model give the 25mg/kg body weight above medicine, and brain cognitive function is significantly improved.

SAMP8 average life is 299 days, and the 4-6 monthly age reaches ripe after date and occurs obviously old and feeblely, is the animal model of relevant disease such as research senile dementia.AD is the modal type of senile dementia, and research [Sun Liang etc. clinical neurology magazine, 2008; 21:91.] find; Can find that before developing into AD the scoring of the simple and easy mental status examination of patient (MMSE) scale is starkly lower than the normal control group, wherein the decline of spatial orientation and delay memory score is more obvious, and its late period is except that having cognitive dysfunction; Also with mental symptom, this and SAMP8 show closely similar.

The Morris water maze is that existing application is widely used in the test method that detects the memory of animal space learning.Originally discover that in the orientation navigation test, the medication therapy groups mice finds the average latency of platform significantly to be shorter than matched group (all P＜0.05).The directed learning ability that shows the medication therapy groups mice all obviously improves than control group mice.In the space exploration test, the medication therapy groups mice is passed through the number of times of original platform position than matched group showed increased (P＜0.05).The spatial memory ability that shows the medication therapy groups mice is improved.This Drug therapy/intervention of this result of study explanation can improve the space learning ability of SAMP8, finds that its behavioristics's ability significantly improves.

In a word, medicine shows that to the experiment that APP expresses influence the above medicine of 5uM has the outstanding effect that reduces the expression of A beta-amyloyd precursor protein.Experiment in the coalition, water maze orientation navigation result of the test shows, gives the 25mg/kg/ body weight/day above medication therapy groups mice, no matter its cognitive function and space learning ability all significantly improve.Pathology detection result shows: brain cognitive function be improved significantly reason be that Drug therapy has reduced the A amyloid beta in the cerebral tissue.Thus, prove that this Drug therapy of 25-75mg/kg/ body weight/day has the curative effect of treatment senile dementia.At these as a result on the basis; Further inference can be thought: this medicine is in to human senile dementia patient treatment; Also can produce similar therapeutic effect, though effective dose still will be groped, our experiment has drawn the scope roughly of medicine for treatment effective dose.

Claims (9)

1. a chemical compound or its pharmaceutically acceptable salt are in the application of control senile dementia, and it is characterized in that: said chemical compound is N'-{5-[acetyl group (hydroxyl) amine] pentyl }-N-[5-({ 4-[(5-aminopentane base) (hydroxyl) amine]-4-ketone-bytyry } amine) pentyl]- N-hydroxy succinic acid amide.

2. application according to claim 1 is characterized in that: described chemical compound or its pharmaceutically acceptable salt can be used as medicine and are used to prevent and treat senile dementia.

3. application according to claim 1 is characterized in that, described salt is mesylate.

4. application according to claim 1; It is characterized in that: said chemical compound or its pharmaceutically acceptable salt see through blood brain barrier and heavy metal is combined into chelate; Suppress amyloid generation in the brain; Blocking-up amyloid polymerization and accumulation are dissolved established amyloid speckle and are promoted the amyloid degraded.

5. application according to claim 4 is characterized in that: described heavy metal is more than one in ferrum, copper, aluminum, the zinc.

6. application according to claim 1 is characterized in that: during medication, can in said chemical compound or its pharmaceutically acceptable salt, add acceptable carrier or solvent on the materia medica.

7. application according to claim 6 is characterized in that, described solvent is that normal saline, distilled water or mass concentration are the glucose of 5-10%.

8. application according to claim 6; It is characterized in that described carrier is more than one in starch, sucrose, mannitol, silicon derivative, carboxymethyl cellulose, gelatin, polyvinylpyrrolidone, glycerol, carboxymethyl starch sodium, crospovidone, low-substituted hydroxypropyl cellulose, paraffin, quaternary ammonium compound, hexadecanol, glyceryl monostearate, Kaolin, bentonite, calcium stearate, magnesium stearate, micropowder silica gel, the Pulvis Talci.

9. application according to claim 1 is characterized in that: the using dosage of said chemical compound or its pharmaceutically acceptable salt is the 2-80mg/kg body weight/day.

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