CN106831573B - (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound, preparation method and applications - Google Patents

Abstract

The invention belongs to pharmaceutical technology fields, are related to a kind of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, preparation method and applications, and structure is shown in formula I,.(N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound of the invention has good butyrylcholine esterase inhibitory activity, significant inhibition A β in vitro experiment_1‑42Aggregation activity; antioxidant activity and there is preferable neuroprotective activity to the PC12 cellular damage of hydrogen peroxide-induced; show compound (N-1; 2; 3,4- tetrahydro isoquinolyls)-asafoetide amide (I) is multiple target point inhibitor, the effect of preferable treatment Alzheimer disease is further shown in vivo experiment; and toxicity is lower, has good potential applicability in clinical practice.

Description

(N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound, preparation method and its Using

Technical field

The invention belongs to pharmaceutical technology fields, and in particular to a kind of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide Compound, preparation method and applications.

Background technique

Alzheimer's disease (Alzheimer's disease, AD, senile dementia) is that a kind of recognized with progressive hinders Hinder with memory damage based on central nervous system degenerative disease, with rapidly aging, the elderly population of population in the world Health problem have become current social significant problem in the urgent need to address.Alzheimer's disease is disease incidence in the elderly One of with the highest disease of lethality.Alzheimer's disease international association (Alzheimer's disease International, ADI) " 2015 global Alzheimer's diseases report " of publication point out that the whole world had had more than 4600 in 2015 Ten thousand people suffer from dementia, it was predicted that the year two thousand fifty, the whole world will have 1.315 hundred million populations by dull-witted puzzlement, wherein China is dull-witted The disease incidence of disease patient has reached 6.61%.With the extension of existent age per capita, this disease has developed into society and health care system The main burden of system, and heavy spirit and economic pressures are brought for society, patient and family members.Approved is for controlling at present The drug for treating light/moderate AD has acetylcholinesterase (AChE) inhibitor, and N- methyl D-asparagus fern for severe AD treatment Propylhomoserin (NMDA) receptor antagonist, but clinical use show these drugs can by improve patient's body levels of acetylcholine or Person inhibits the exitotoxicity of excitatory amino acid to alleviate AD symptom, but not can effectively prevent or reverse the course of disease, but also can draw The serious toxic side effects such as illusion, misunderstanding, dizziness, headache, nausea, hepatotoxicity, loss of appetite and stool frequency are played, because And long-term efficacy is not satisfactory.Therefore, clinically there is an urgent need to research and develop the AD therapeutic agent with novel mechanism of action.

In recent years, with constantly illustrating to AD pathogenesis, it is found that the occurrence and development of AD have multimachine system, multifactor It is the characteristics of effect, again interrelated between different mechanisms to influence each other, constitute complicated network during AD occurrence and development Regulator control system.Based on the above results, the clinical efficacy generated for the drug of single definite target spot is not appropriate for the complexity with AD Essence, researcher propose that " multiple target point targeted drug " (Multitarget-directed Ligands, MTDLs) strategy is recognized To be a kind of effective ways for researching and developing anti-neurodegenerative disease drug.It is somebody's turn to do " multiple target point targeted drug " and refers to single chemical entities Multiple target spots in disease network are acted on simultaneously, synergistic effect can produce to the effect of each target spot, and gross effect is made to be greater than each list The sum of effect, such medicine are also referred to as " Multifunctional " or " Multipotential " drug.Up to the present, although The advantage of multiple target point is that clearly, but multiple target spot functions how to combine in the same molecule and most suitable therapy target Selection is still a key point.

With the development of the process of AD, acetylcholinesterase (AChE) level gradually lowers, and butyrylcholine esterase (BuChE) activity increases the 165% of normal value.In the middle severe stage of AD, BuChE replaces AChE to carry out hydrolyse acetylcholine (ACh), the inhibition of BuChE may be more effective in AD treatment.In addition, cascading hypothesis, intracerebral according to beta-amyloid protein The generation and aggregation of oligomer A β has caused pathogenic generation, has eventually led to neuron loss and dementia, A β is able to enter line Plastochondria induced oxidation stress, simultaneous oxidation stress be present in AD patient's intracerebral, and promote A β toxicity by the generation of free radical, into One step deteriorates AD process (Proc. Natl. Acad. Sci. U. S. A. 2005,102,17213-17218. J. Med. Chem. 2016,59,7683-7689.).Thus, it is found that with inhibiting butyrylcholine esterase, A beta-aggregation and having anti- The neuroprotective agent of oxidation activity may be AD, and especially middle severe AD brings dawn.

Summary of the invention

In view of this, technical problem to be solved by the invention is to provide a kind of (N-1,2,3,4- tetrahydro isoquinolyls)- Asafoetide amide compound treats Alzheimer's disease for multiple target point and provides new thinking.

In order to solve the above technical problems, the technical scheme adopted by the invention is that:

(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, chemical structure are shown in formula I:

, in formula, Me represents methyl.

The present invention also provides the preparation method of above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, The following steps are included:

, In formula, Me represents methyl,

Under solvent and condensing agent existence condition condensation reaction occurs for ferulic acid (1) and 1,2,3,4- tetrahydroisoquinolines (2), Compound (I) is obtained,

The solvent is one of methylene chloride, tetrahydrofuran and toluene or a variety of,

The condensing agent is one of EDCI, HOBT, DCC, DMAP and Ka Te condensation reagent or a variety of.

The present invention also provides the prodrug of above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, foundations The present invention, prodrug are the derivatives of above compound, in physiological conditions (such as passing through metabolism, solvolysis or other means) It is converted to corresponding biologically active form.Pharmaceutically acceptable prodrug, which refers to, to be had by solvolysis or in physiology Under the conditions of can be converted to amino, hydroxyl, carboxyl etc. group compound.As the group for forming prodrug, public affairs can be enumerated Know document such as Prog.Med., 5,2157-2161 (1985) and " exploitations of pharmaceuticals ", Guang Chuan bookstore, 1990, the 7th Volume, the group of 163-198 pages of record.

The present invention also provides above-mentioned (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compounds can pharmaceutically connect The hydrate received.

The present invention also provides above-mentioned (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound or its pharmaceutically may be used Purposes of the hydrate of receiving in preparation treatment neurodegenerative disease drug.Preferably, the neurodegenerative disease is Alzheimer's disease.

The present invention also provides a kind of pharmaceutical compositions, including a effective amount of above-mentioned (N-1,2,3,4- tetrahydroisoquinolines Base)-asafoetide amide compound or its pharmaceutically acceptable hydrate.

Preferably, the dosage form of aforementioned pharmaceutical compositions be oral quick disintegrating tablet, oral cavity compound preparation, oral sustained-release preparation, Depot long-acting injection or Percutaneously administrable preparation.

The present invention also provides purposes of the aforementioned pharmaceutical compositions in preparation treatment neurodegenerative disease drug, especially Ground, the neurodegenerative disease are Alzheimer's disease.

Compared with prior art, the invention has the benefit that

(N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound of the invention has significant suppression to butyrylcholine esterase System activity, IC₅₀It is 3.4 μM, inhibitory activity of the more existing widely used anti-AD medicine donepezil to butyrylcholine esterase (IC₅₀It is 4.76 μM) it is high；And the inhibitory activity to acetylcholinesterase is significantly higher than to the inhibitory activity of butyrylcholine esterase, it says Bright the compounds of this invention has selective inhibitory to butyrylcholine esterase.

(N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound of the invention is to A β_1-42Self assemble all has aobvious Write inhibitory activity, inhibiting rate is 61.1% (25 μM), is significantly higher than existing anti-AD medicine donepezil (less than 5%).

The present invention (N- 1,2,3,4- tetrahydro isoquinolyl) antioxidant activity of-asafoetide amide compound is Trolox 1.1 times, there is preferable antioxidant activity.

(N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound of the invention has significant neuroprotection.

Provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) experiment in vitro show it is good Butyrylcholine esterase inhibitory activity, significant inhibition A β_1-42Aggregation activity, antioxidant activity and the PC12 to hydrogen peroxide-induced Cellular damage has preferable neuroprotective activity, shows compound (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide (I) It is a multiple target point inhibitor.The effect of preferable treatment Alzheimer disease is shown in experiment in vivo, and toxicity is lower, has Good potential applicability in clinical practice.

Detailed description of the invention

Present invention will be described in further detail below with reference to the accompanying drawings:

Fig. 1: the PC12 cytotoxicity assay result of the compounds of this invention (I)；

Fig. 2: the compounds of this invention (I) is to H₂O₂The protective effect measurement result of the PC12 cellular damage of induction；

Fig. 3: the compounds of this invention (I) is to the memory representational role Disorder Model evaluation of hyoscine induced mice.

Specific embodiment

For a better understanding of the present invention, the contents of the present invention, but this hair are further fairly set out below with reference to embodiment Bright protection content is not limited solely to the following examples.In the following description, give a large amount of concrete details so as to More thorough understanding of the invention is provided.It will be apparent, however, to one skilled in the art that the present invention can be with It is carried out without one or more of these details.

Embodiment 1-10

The preparation method of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, comprising the following steps:

, In formula, Me represents methyl,

Ferulic acid (1), condensing agent and solvent are added in reaction flask, 1,2,3,4- tetrahydroisoquinolines are stirring evenly and then adding into (2), it finishes, is stirred to react under temperature T n hours, TLC monitoring；After reaction, water is added in residue in evaporating solvent under reduced pressure, It is extracted with dichloromethane, organic layer is washed after merging with saturated sodium-chloride water solution, and anhydrous sodium sulfate dries, filters, and filtrate subtracts Pressure is evaporated off solvent, and residue purifies (eluant, eluent: petroleum ether/acetone=20/1) with silica gel column chromatography, obtain target product (N-1,2, 3,4- tetrahydro isoquinolyl)-asafoetide amide (I).

(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide (I) is white solid, and fusing point is 181.6 ~ 183.7 °C, is received Rate is 30% ~ 85%, and chemical structure passes through¹H NMR、¹³C NMR and ESI-MS confirmation.

Wherein, the solvent is one of methylene chloride, tetrahydrofuran and toluene or a variety of；The condensing agent is One of EDCI, HOBT, DCC, DMAP and Ka Te condensation reagent is a variety of；The ferulic acid (1): 1,2,3,4- Tetrahydroisoquinoli- Quinoline (2): the molar feed ratio of condensing agent is 1:1.0 ~ 10.0:1.0 ~ 10.0；Reaction temperature be 0 ~ 105 DEG C, the reaction time be 1 ~ 72 hours.

In the present invention, EDCI:1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride, chemical formula is C₈H₁₈ClN₃；HOBT:1- hydroxybenzotriazole, chemical formula C₆H₅N₃O；DCC;Dicyclohexylcarbodiimide, chemical formula are C₁₃H₂₂N₂；DMAP:4- dimethylamino naphthyridine, chemical formula C₇H₁₀N₂；Block special condensation reagent: (the front three ammonia of benzotriazole -1- three Base)-trifluoro phosphate, chemical formula C₁₂H₂₂F₆N₆OP₂。

The concrete technology condition of embodiment 1-10 is shown in Table 1.

1 present invention process condition of table

。

1 gained (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) of the embodiment of the present invention¹H NMR、¹³C NMR With ESI-MS testing result are as follows:

¹H NMR (400 MHz, CDCl₃) δ 7.66 (d,J=15.2 Hz, 1H, C=CH), 7.24-7.12 (m, 5H, 5 × Ar-H), 7.16 (d,J=1.6 Hz, 1H, Ar-H), 6.93 (d,J=8.0 Hz, 1H, Ar-H), 6.80 (d,J =15.2 Hz, 1H, C=CH), 6.04 (s, 1H, OH), 4.84 (s, 2H, phCH₂N), 3.94 (s, 3H, OCH₃), 3.88 (s, 2H, phCH₂), 3.97-2.91 (m, 2H, NCH₂), 3.80-3.74 (m, 4H, 2 × NCH₂).

¹³C NMR (100 MHz, CDCl₃) 166.24,147.42,146.74,143.06,134.25,133.70, 128.97,128.26,127.81,126.76,126.14,121.90,114.79 (2C), 110.02,56.02,43.60, 29.74.

MS (ESI) m/z: 310.1 [M + H]⁺.

The present invention also provides a kind of pharmaceutical compositions for treating neurodegenerative disease, including a effective amount of above-mentioned treatment Neurodegenerative disease drug or its pharmaceutically acceptable hydrate.Described pharmaceutical composition can be further containing a kind of or more Kind pharmaceutically acceptable carrier or excipient." effective quantity ", which refers to, to be caused researcher or the targeted tissue of doctor, is The biology or medicine of system or animal react drug or medicament amount；" composition " refer to by by more than one substances or The product that component mixes；" pharmaceutically acceptable carrier " refers to pharmaceutically acceptable substance, composition or load Body, such as: liquid or solid filler, diluent, excipient, solvent or packing substance, they carry or transport certain chemicals Matter.The pharmaceutically acceptable excipient can for example be enumerated: lactose, glucose, starch, sucrose, microcrystalline cellulose, licorice powder End, mannitol, sodium bicarbonate, calcium phosphate, calcium sulfate.

Aforementioned pharmaceutical compositions can be further oral quick disintegrating tablet, oral cavity compound preparation, oral sustained-release preparation, depot Long-acting injection or Percutaneously administrable preparation.

Embodiment 11

(1) acetylcholinesterase and butyrylcholine esterase inhibitory activity

1.0 mmol/L acetylthiocholine iodides are sequentially added into 96 orifice plates or thio BuCh (is purchased from Sigma company) 30 μ L, the 40 μ L of PBS buffer solution of pH=8.0,20 μ L of testing compound solution (DMSO content is less than 1%) and 10 μ L acetylcholinesterases (EeAChE) or butyrylcholine esterase (equine serum BuChE,eqBuChE) (0.045U, Purchased from Sigma company), addition terminates after mixing, 37 °C of 15 min of incubation, 5 that mass fraction is 0.2% are added into each hole, The 30 μ L colour developing of thio-bis- (2- nitro) benzoic acid of 5'- bis- (DTNB is purchased from Sigma company) solution, measures 405 nm with microplate reader The OD value (OD value) for locating each hole, compared with the blank well that sample to be tested is not added, calculating inhibiting rate of the compound to enzyme, [enzyme presses down Rate processed=(1- sample sets OD value/blank group OD value) × 100%]；Five to six concentration for selecting compound measure the inhibition of its enzyme Rate, and with the inhibiting rate linear regression of the negative logarithm of the compound molar concentration and enzyme, acquire molar concentration when 50% inhibiting rate The as IC of the compound₅₀。

See Table 2 for details for experimental result.

Table 2 (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) cholinesterase inhibition

,

Table 2 the result shows that, it is provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) is to butyryl Cholinesterase has remarkable inhibiting activity, IC₅₀It is 3.4 μM；More existing widely used anti-AD medicine donepezil is to butyryl gallbladder Inhibitory activity (the IC of alkali esterase₅₀It is 4.76 μM) it is high.Also, (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) is to fourth Inhibitory activity (the IC of acetylcholinesterase₅₀It is 3.4 μM) it is significantly higher than to acetylcholinesterase (IC₅₀Be 5.6 μM) inhibition it is living Property, illustrate that compound provided by the present invention (I) has selective inhibitory to butyrylcholine esterase.

(2) to Aβ _1-42The inhibitory activity of self assemble

Reference literature (Sang, Z.P. et al. Eur. J. Med. Chem. 2015,94,348-366) is reported Method is measured, it may be assumed that pretreated Aβ _1-42It is made into stock solution with DMSO, is diluted using the preceding PBS buffer solution with pH7.4 To 50 μM；Untested compound is made into 2.5mM stock solution with DMSO, using it is preceding be diluted to the PBS buffer solution of pH7.4 it is corresponding dense Degree, takes the A of 20 μ Lβ _1-42The testing compound solution of+20 μ L of solution, 20 μ L Aβ _1-42+ 20 μ L PBS buffer solution of solution (contains 2% DMSO), 20 μ L PBS buffer solution (contain 2% DMSO)+20 μ L PBS buffer solution (containing 25% DMSO) are in 96 orifice plate of black In, compound and Aβ _1-42Ultimate density be 25 μM.37 °C of 24 h of incubation, are then added 160 μ L and contain 5 μM of thioflavines The glycine-NaOH buffer (pH=8.5) of 50 mM of T uses Varioskan Flash Multimode after shaking 5s immediately Reader multi-function microplate reader measures fluorescent value under 446 nm excitation wavelengths and 490 nm launch wavelengths；Aβ _1-42+ test compounds The fluorescent value of object is recorded as IF_i, Aβ _1-42The fluorescent value of+PBS buffer solution is recorded as IF_c, contain only the fluorescent value note of PBS buffer solution Record is IF₀, A is inhibited by compoundβ _1-42The inhibiting rate calculation formula of self assemble are as follows: 100- (IF_i-IF₀)/(IF_c-IF₀)× 100；Each each two multiple holes of concentration mensuration of compound；Using curcumin as positive control.

See Table 3 for details for experimental result.

Table 3 (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) is to Aβ _1-42The inhibitory activity of self assemble is tested

,

^a 25 μM of inhibitor concentration, and the of Inhibition was determined at mean ± SD of the 3 independent experiments.

Table 3 is the result shows that compound (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) has preferable inhibition A β_1-42Aggregation activity, inhibiting rate 61.1%；The inhibiting rate of positive drug curcumin is 56.8%；Widely used anti-AD medicine mostly how piperazine Together to A β under 25 μM of concentration_1-42The inhibiting rate of self assemble is less than 5%.

(3) Antioxidative Activity Determination (ORAC-FL method)

Reagent and instrument: 6- hydroxyl -2,5, (trolox changes 7,8- tetramethyl primary colours alkane -2- carboxylic acids purchased from the uncommon love (Shanghai) of ladder At industrial development Co., Ltd) solution of 10-80 μm of ol/L is made into the PBS buffer solution (pH7.4) of 75mM；Fluorescein (fluorescein, purchased from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder) is matched with the PBS buffer solution (pH7.4) of 75 mM At the solution of 250 nmol/L；(AAPH has 2,2 '-azo diisobutyl amidine dihydrochlorides purchased from splendid remote chemistry scientific and technological (Shanghai) Limit company) using the preceding solution that 40 mmol/L are made into the PBS buffer solution (pH7.4) of 75 mM；

Microplate reader is Varioskan Flash Multimode Reader(Thermo Scientific).

Measurement experiment method: the 20 μ L of compound solution and fluorescein of 50 or 10 μm of ol/L are added into 96 orifice plate of black 120 μ L of solution is mixed, 37 °C of 15 min of incubation, and 60 μ L of AAPH solution is added, makes every 200 μ L of hole total volume, is mixed, It is immediately placed in Varioskan Flash Multimode Reader instrument, in 485 nm excitation wavelengths and 535 nm transmitted waves Long lower measurement first order fluorescence value, 90 min of METHOD FOR CONTINUOUS DETERMINATION per minute calculate area under fluorescence decay curve by instrument automatically AUC.Wherein using the trolox of 1-8 μm of ol/L as standard, sample to be tested is not added as blank.The antioxidant activity of compound Results expression is the equivalent of trolox, calculation formula are as follows: [(AUC Sample-AUC blank)/(AUC Trolox-AUC blank)][(concentration of Trolox/concentration of sample)].Each compound measures every time 3 multiple holes, every group of experiment are independent in triplicate.

See Table 4 for details for experimental result.

Table 4 (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) Antioxidative Activity Determination

Table 4 the result shows that, it is provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) antioxygen Change 1.1 times that activity is Trolox, there is preferable antioxidant activity.

(4) cytotoxicity assay of the compound (I) to PC12 cell

DMEM culture solution of the PC12 cell containing 10% fetal calf serum, with density for 4 × 10⁴A/ml is inoculated in the culture of 96 holes On plate, inoculation volume is 100 holes μ l/, is subsequently placed into containing 5%CO₂37 °C of constant incubators in culture.PC12 cell culture 24 After hour, compound (I) (final concentration of 100 μM, 10 μM, 1 μM) 10 holes μ L/ of various concentration, constant temperature are added in administration group 3- (4,5- dimethylthiazole -2- base) -2,5- diphenyltetrazolium bromide bromide of 5 mg/mL is added after cultivating 24 h, in each group (MTT) 100 hole μ L/ carries out living cells dyeing.After 3 hours, 100 hole μ L/ of 100%DMSO terminate liquid is added in each group, fills Dissolution is divided to mix.The OD value of each group is measured under the wavelength of 590 nm.Each compound is respectively with 100 μM, 10 μM, 1 μM of test 3 times as a result, with Duncan ' s test method statistic, with control group for 100%, administration class value is indicated with the percentage of control group. Inhibiting rate=(control-compound)/control * 100%.

Measurement result is detailed in Fig. 1, shows that compound (I) has lower cytotoxicity, possesses the treatment model an of safety It encloses.

(5) compound is to H₂O₂The protective effect of the PC12 cellular damage of induction is screened

DMEM culture solution of the PC12 cell containing 10% calf serum, with 1 × 10⁵A/mL density is inoculated in 96 well culture plates On, inoculation volume is the hole 100mL/, is subsequently placed into containing 5%CO₂37 DEG C of constant incubators in culture.After culture 24 hours, administration In group plus compound (final concentration of 10 μM, 1 μM) hole 10mL/ of respective concentration, preincubate 2 hours (control group and damage components Not plus 10 μ L/ hole PBS, its volume is made to keep equal).After PC12 cell incubation 2 hours, add respectively in administration group and damage group Enter 100 μ Μ H₂O₂It damages 10 hole μ L/ of agent (control group adds 10 μ L/ hole PBS), after 30 minutes, changes the culture solution of each group into nothing The culture solution of calf serum continues to be put into culture 24 hours in constant incubator, and nutrient solution volume thinks 100 holes μ L/.Continue to train After supporting 24 hours, 5mg/mL is added in each group, and 100 hole μ L/ MTT carries out living cells dyeing.After 3 hours, it is added in each group 100 hole μ L/ of 100%DMSO terminate liquid, sufficiently dissolution mix.The OD value of each group, test result weight are measured under the wavelength of 490 nm 3 times multiple, with Duncan ' s test method statistic, each group numerical value is expressed as mean ± S.E.M., with control group for 100%, administration Group and damage class value are indicated with the percentage of control group.

Measurement result is detailed in Fig. 2, Fig. 2 show compound (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) is right H₂O₂The PC12 cellular damage of induction has significant neuroprotection.

(6) acute toxicity test of compound (I)

Test material: experimental animal is SPF Kunming mice, is provided by Sichuan Provincial Academy of Traditional Chinese Medicine, and production is qualified Card SCXK (river) 2008-19.Animal feeding is in Sichuan Provincial Academy of Traditional Chinese Medicine pharmacological toxicology research institute SPF barrier system. 20~22 DEG C of room temperature, relative humidity 40%-70% or so, illuminate 12 hours bright, 12 hours dark, free water.Quan Ying Pellet is supported to be provided by Sichuan Provincial Academy of Traditional Chinese Medicine Experimental Animal Center.

Experimental method: animal is grouped at random: take mouse 40 of 18 ~ 22g of SPF grade, half male and half female, and adaptable fed two days Afterwards, 4 groups are randomly divided by weight.After being deprived of food but not water 15h, respectively stomach-filling compound (I) 1000 mg/kg, 500 mg/kg, 250 mg/kg, 100 mg/kg, tetra- dosage groups, taking administered volume is 0.4 mL/10g, and each group is administered once, and is seen within continuous 14 days The death condition for examining and recording each animal is analyzed using Bliss statistical software.It was found that each group mouse do not occur hair hold up, It is slow in action, closes one's eyes and breathes acceleration and the phenomena of mortality.Measurement result show SPF Kunming mice through (N-1,2,3,4- Tetrahydro isoquinolyl) after the processing of-asafoetide amide (I), does not occur anxious poison and the death rate, do not occur hair yet and hold up, be slow in action, close Phenomena such as eye and breathing accelerate, it is nontoxic for showing compound, and maximal tolerance dose is 1000 mg/kg.

(7) zoopery-diving tower passive avoidance test

Reagent and instrument: donepezil is purchased from Eisai China Inc.；Hyoscine is purchased from J&K Scienti c；18- The Kunming mouse of 22g is purchased from Sichuan scientific tcm institute Experimental Animal Center (quality certification number: SCXK-Sichuan 2008-19)；It is dynamic Object is raised in Sichuan Provincial Academy of Traditional Chinese Medicine pharmacological toxicology research institute SPF barrier system.12 h illumination of feeding environment/12 h are black Dark alternating, environment temperature are controlled in 20-22 °C, and humid control is in 50-60%.Full nutritious particle feed is by traditional Chinese medicine section, Sichuan Province Institute's Experimental Animal Center provides.Mouse diving tower instrument (model ZXC-5Q) is raw by Shandong Academy of Medical Sciences's maintenance of equipment supply station It produces

Experimental method: 60 mouse, 18 ~ 22 g, half male and half female are randomly divided into 6 groups by weight, i.e., blank control group, Model control group, donepezil group (5.0mgkg^-1), compound (I) high dose group (10.0 mgkg^-1), compound (I) Middle dose group (5.0mgkg^-1), compound (I) low dose group (2.5mgkg^-1).Every group of mouse is divided up and down by dosage Noon administration, successive administration 3 times, 50 min carry out modeling after the last administration, other each groups are injected intraperitoneally in addition to blank control group 3 mgkg of hyoscine^-1, successive administration 24 days.20 min carry out step dow n test training after modeling, and animal is put into reaction chamber 3 min are adapted to, pass to 36 V alternating currents, 5 min of training immediately after, and record each mouse by the number (mistake to shock by electricity Number), and thus it is used as school grade.It is tested, every 5 min of mouse assay, is recorded by the animal shocked by electricity after 24 h Number and first time jump off the errors number in the incubation period and 5 min of platform, as a result carry out statistical analysis, all data are equal It is indicated with mean value ± standard error (Stand error, S.E.).It is analyzed using SPSS11.5 software, the neat selection Dan Yin of variance Plain variance analysis (One-way ANOVA).Measurement data compares using one-way analysis of variance, and the comparison of each group mean uses t It examines, as a result sees Fig. 3 respectively.

Refering to Fig. 3, the results showed that the present invention (N- 1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide (I) is to eastern Liang Henbane alkali induced mice memory representational role obstacle all has the effect of being obviously improved.

Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, this field is common Other modifications or equivalent replacement that technical staff makes technical solution of the present invention, without departing from technical solution of the present invention Spirit and scope, be intended to be within the scope of the claims of the invention.

Claims (8)

1. (1. N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, it is characterised in that: chemical structure is shown in formula I:

   。
2. 2. a kind of preparation method of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, which is characterized in that including with Lower step:

   ,

   Ferulic acid (1) and 1,2,3,4- tetrahydroisoquinolines (2) occur condensation reaction under solvent and condensing agent existence condition, obtain (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound (I).
3. 3. the preparation method of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound as claimed in claim 2, special Sign is: the solvent be one of methylene chloride, tetrahydrofuran, toluene or a variety of, the condensing agent be EDCI, HOBT, One of DCC, DMAP and Ka Te condensation reagent is a variety of.
4. 4. (N-1,2,3,4- tetrahydro isoquinolyl)-asafoetide amide compound as described in claim 1 is in preparation treatment nerve Application in degenerative disease drug.
5. 5. a kind of pharmaceutical composition, it is characterised in that: including it is a effective amount of it is as claimed in claim 1 or 2 (N-1,2,3,4- tetra- Hydrogen isoquinoline base)-asafoetide amide compound.
6. 6. pharmaceutical composition according to claim 5, it is characterised in that: its dosage form is oral quick disintegrating tablet, oral cavity compound system Agent, oral sustained-release preparation, depot long-acting injection or Percutaneously administrable preparation.
7. 7. application of the pharmaceutical composition as claimed in claim 5 in preparation treatment Alzheimer's disease drug.
8. 8. application of the pharmaceutical composition as claimed in claim 6 in preparation treatment Alzheimer's disease drug.