CN106831573A - (tetrahydro isoquinolyls of N 1,2,3,4) asafoetide amide compound, preparation method and applications - Google Patents
(tetrahydro isoquinolyls of N 1,2,3,4) asafoetide amide compound, preparation method and applications Download PDFInfo
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- C07D217/02—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with only hydrogen atoms or radicals containing only carbon and hydrogen atoms, directly attached to carbon atoms of the nitrogen-containing ring; Alkylene-bis-isoquinolines
- C07D217/06—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with only hydrogen atoms or radicals containing only carbon and hydrogen atoms, directly attached to carbon atoms of the nitrogen-containing ring; Alkylene-bis-isoquinolines with the ring nitrogen atom acylated by carboxylic or carbonic acids, or with sulfur or nitrogen analogues thereof, e.g. carbamates
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Abstract
The invention belongs to pharmaceutical technology field, be related to a kind of (tetrahydro isoquinolyls of N 1,2,3,4) asafoetide amide compound, preparation method and applications, structure shown in formula I,.(tetrahydro isoquinolyls of N 1,2,3,4) asafoetide amide compound of the invention has good butyrylcholine esterase inhibitory activity, significant suppression A β in testing in vitro1‑42Aggregation activity, antioxidation activity and the PC12 cellular damages to hydrogen peroxide-induced have preferable neuroprotective activity, show compound (tetrahydro isoquinolyls of N 1,2,3,4) asafoetide acid amides（I）It is a Mutiple Targets inhibitor, the effect of preferably treatment Alzheimer disease is shown in further testing in vivo, and toxicity is relatively low, possesses good potential applicability in clinical practice.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to a kind of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides
Compound, preparation method and applications.
Background technology
Alzheimer's disease（Alzheimer's disease, AD, senile dementia）It is that one kind hinders so that progressive is cognitive
Hinder with memory infringement based on central nervous system degenerative disease, with the rapidly aging of population in the world, elderly population
Health problem have become current social significant problem in the urgent need to address.Alzheimer's disease is the incidence of disease in the elderly
One of with fatal rate highest disease.Alzheimer's disease international association（Alzheimer's disease
International, ADI）Issue《2015 global Alzheimer's disease reports》Point out, the whole world has had more than 4600 within 2015
Ten thousand people suffer from dementia, it was predicted that to the year two thousand fifty, the whole world will have 1.315 hundred million populations to be perplexed by dementia, wherein China is dull-witted
The incidence of disease 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 for society, patient and family members bring heavy spirit and economic pressures.Current approved is used to control
The medicine for treating light/moderate AD has acetylcholinesterase（AChE）Inhibitor, and for the N- methyl Ds-asparagus fern of severe AD treatments
Propylhomoserin（NMDA）Receptor antagonist, but Clinical practice shows, these medicines can by improve in patient's body levels of acetylcholine or
The exitotoxicity that person suppresses excitatory amino acid alleviates AD symptoms, but can not effectively prevent or reverse the course of disease, but also can draw
The serious toxic and side effect such as illusion, misunderstanding, dizziness, headache, nausea, hepatotoxicity, poor appetite and stool frequency is played, because
And long-term efficacy is not satisfactory.Therefore, clinically in the urgent need to AD medicine of the research and development with new mechanism of action.
In recent years, with constantly illustrating to AD pathogenesis, finding the occurrence and development of AD has many mechanism, multifactor
The characteristics of effect, the again interrelated network for influencing each other, constituting complexity during AD occurrence and development between different mechanisms
Regulator control system.Based on the above results, the clinical efficacy produced for the medicine of single definite target spot is not appropriate for the complexity with AD
Essence, researcher proposes " Mutiple Targets targeted drug "（Multitarget-directed Ligands, MTDLs）Strategy is recognized
To be a kind of effective ways for researching and developing anti-nerve degenerative diseases medicine.It refers to single chemical entities to be somebody's turn to do " Mutiple Targets targeted drug "
The multiple target spots in disease network are acted on simultaneously, the effect to each target spot can produce cooperative effect, gross effect is more than each list
Effect sum, such medicine is also referred to as " Multifunctional " or " Multipotential " medicine.Up to the present, although
The advantage of Mutiple Targets is that clearly, but how multiple target spot function combines in 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 substitutions AChE carrys out hydrolyse acetylcholine
（ACh）, the suppression of BuChE may be more effective in AD treatments.In addition, cascading hypothesis, intracerebral according to amyloid-beta
The generation and aggregation of oligomer A β have triggered pathogenic generation, have ultimately resulted in neuron loss and dementia, and A β can enter line
Plastochondria induced oxidation stress, simultaneous oxidation stress be present in AD patient's intracerebral, and pass through the generation of free radical and promote A β toxicity, enter
One step deteriorates AD processes（Proc. Natl. Acad. Sci. U. S. A. 2005, 102, 17213-17218. J.
Med. Chem. 2016, 59, 7683-7689.）.Thus, it is found that have suppressing butyrylcholine esterase, A beta peptide aggregations and with anti-
The neuroprotective agent of oxidation activity may bring dawn for AD, especially middle severe AD.
The content of the invention
In view of this, the technical problems to be solved by the invention be to provide a kind of (N-1,2,3,4- tetrahydro isoquinolyls)-
Asafoetide amide compound, for Mutiple Targets treatment Alzheimer's provides new thinking.
In order to solve the above technical problems, the technical solution adopted in the present invention is：
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, chemical constitution is shown in formula I：
,
In formula, Me represents methyl.
Present invention also offers the preparation method of above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound,
Comprise the following steps：
,
In formula, Me represents methyl,
Forulic acid（1）With 1,2,3,4- tetrahydroisoquinolines（2）There is condensation reaction under solvent and condensing agent existence condition, obtain
Compound（I）,
The solvent is one or more in dichloromethane, tetrahydrofuran and toluene,
The condensing agent is one or more in EDCI, HOBT, DCC, DMAP and Ka Te condensation reagent.
Present invention also offers the prodrug of above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, foundation
The present invention, prodrug is the derivative of above-claimed cpd, in physiological conditions（For example by metabolism, solvolysis or other manner）
It is converted to corresponding biologically active form.Pharmaceutically acceptable prodrug refers to have 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 bookstores, the 7th in 1990
Volume, the 163-198 pages of group of record.
Can pharmaceutically be connect present invention also offers above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound
The hydrate received.
Present invention also offers above-mentioned (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound or its pharmaceutically may be used
Purposes of the hydrate of receiving in treatment nerve degenerative diseases medicine is prepared.Preferably, the nerve degenerative diseases are
Alzheimer's.
Present invention also offers a kind of pharmaceutical composition, including effective dose above-mentioned (N-1,2,3,4- tetrahydroisoquinolines
Base)-asafoetide amide compound or its pharmaceutically acceptable hydrate.
Preferably, the formulation 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 purposes in treating nerve degenerative diseases medicine is being prepared present invention also offers aforementioned pharmaceutical compositions, especially
Ground, the nerve degenerative diseases are Alzheimer's.
Compared with prior art, beneficial effects of the present invention are：
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound of the invention has to butyrylcholine esterase and significantly inhibits work
Property, its IC50It is 3.4 μM, inhibitory activity of the more existing widely used anti-AD medicines donepezil to butyrylcholine esterase（IC50For
4.76μM）It is high；And the inhibitory activity to acetylcholinesterase is significantly higher than to the inhibitory activity of butyrylcholine esterase, illustrate this hair
Bright compound has selective inhibitory to butyrylcholine esterase.
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound of the invention is to A β1-42Self assemble is respectively provided with aobvious
Inhibitory activity is write, its inhibiting rate is 61.1% (25 μM), is significantly higher than existing anti-AD medicines donepezil（Less than 5%）.
The present invention (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound antioxidation activity for Trolox
1.1 times, with preferable antioxidation activity.
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound of the invention has significant neuroprotection.
Provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）Experiment in vitro shows good
Butyrylcholine esterase inhibitory activity, significant suppression A β1-42Aggregation activity, antioxidation activity and the PC12 to hydrogen peroxide-induced
Cellular damage has preferable neuroprotective activity, shows compound (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）
It is a Mutiple Targets inhibitor.The effect of preferably treatment Alzheimer disease is shown in experiment in vivo, and toxicity is relatively low, possesses
Good potential applicability in clinical practice.
Brief description of the drawings
The present invention is described in further detail below in conjunction with the accompanying drawings：
Fig. 1：The compounds of this invention（I）PC12 CTA results；
Fig. 2：The compounds of this invention（I）To H2O2The protective effect measurement result of the PC12 cellular damages of induction；
Fig. 3：The compounds of this invention（I）Hyoscine induced mice memory representational role Disorder Model is evaluated.
Specific embodiment
For a better understanding of the present invention, present disclosure, but this hair are further fairly set out 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, comprises the following steps：
,
In formula, Me represents methyl,
By forulic acid（1）, during condensing agent and solvent add reaction bulb, be stirring evenly and then adding into 1,2,3,4- tetrahydroisoquinolines（2）,
Finish, stirring reaction n hours under temperature T, TLC monitorings；After reaction terminates, solvent is removed under reduced pressure, water is added in residue, with two
Chloromethanes is extracted, and organic layer is washed after merging with saturated sodium-chloride water solution, anhydrous sodium sulfate drying, and filtering, filtrate decompression is steamed
Except solvent, residue is purified with silica gel column chromatography（Eluant, eluent：Petroleum ether/acetone=20/1）, obtain target product (N-1,2,3,4-
Tetrahydro isoquinolyl)-asafoetide acid amides（I）.
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）It is white solid, fusing point is 181.6 ~ 183.7 °C, is received
Rate is 30% ~ 85%, and its chemical constitution is passed through1H NMR、13C NMR and ESI-MS are confirmed.
Wherein, the solvent is one or more in dichloromethane, tetrahydrofuran and toluene；The condensing agent is
One or more in EDCI, HOBT, DCC, DMAP and Ka Te condensation reagent；The forulic acid（1）：1,2,3,4- Tetrahydroisoquinoli-s
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-propyls) -3- ethyl-carbodiimide hydrochlorides, chemical formula is
C8H18ClN3；HOBT：I-hydroxybenzotriazole, chemical formula is C6H5N3O；DCC;Dicyclohexylcarbodiimide, chemical formula is
C13H22N2；DMAP:DMAP, chemical formula is C7H10N2；The special condensation reagent of card:(the front three ammonia of BTA -1- three
Base)-trifluoro phosphate, chemical formula is C12H22F6N6OP2。
The concrete technology condition of embodiment 1-10 is shown in Table 1.
The present invention process condition of table 1
。
Gained (N-1,2,3,4- the tetrahydro isoquinolyls)-asafoetide acid amides of the embodiment of the present invention 1（I）'s1H NMR、13C NMR
It is with ESI-MS testing results：
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]+.
It is neural present invention also offers a kind of pharmaceutical composition for treating nerve degenerative diseases, including the above-mentioned treatment of effective dose
DD medicine or its pharmaceutically acceptable hydrate.Described pharmaceutical composition can further contain one or more medicine
Acceptable carrier or excipient on." effective dose " refer to cause researcher or the targeted tissue of doctor, system or
Animal it is biological or medical react medicine or medicament amount；" composition " refers to by by more than one materials or component
The product for mixing；" pharmaceutically acceptable carrier " refers to pharmaceutically acceptable material, composition or carrier,
Such as：Liquid or solid filler, diluent, excipient, solvent or packing material, they carry or transport certain chemical substance.
The pharmaceutically acceptable excipient can for example be enumerated：Lactose, glucose, starch, sucrose, microcrystalline cellulose, Radix Glycyrrhizae powder,
Mannitol, sodium acid carbonate, 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
To sequentially adding 1.0 mmol/L acetylthiocholine iodides or thio BuCh in 96 orifice plates（It is purchased from Sigma public
Department）The μ L of PBS 40 of 30 μ L, pH=8.0, the μ L of testing compound solution 20 (DMSO contents are less than 1%) and 10 μ L second
Acetylcholinesterase（EeAChE）Or butyrylcholine esterase (equine serum BuChE,eqBuChE) (0.045U is purchased from
Sigma companies), after addition terminates to mix, 37 °C of 15 min of incubation, to 5, the 5'- bis- that addition mass fraction in each hole is 0.2%
Thio-bis- (2- nitros) benzoic acid (DTNB, purchased from Sigma companies) solution 30 μ L colour developings, determine each at 405 nm with ELIASA
The OD value (OD values) in hole, compares with the blank well for being not added with testing sample, calculates inhibiting rate [enzyme inhibition rate of the compound to enzyme
=(1- sample sets OD values/blank group OD values) × 100%]；Five to six concentration of compound are selected, its enzyme inhibition rate is determined, and
With the inhibiting rate linear regression of the negative logarithm of the compound molar concentration and enzyme, molar concentration when trying to achieve 50% inhibiting rate is
The IC of the compound50。
Experimental result refers to table 2.
Table 2 (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）Cholinesterase inhibition
,
The result of table 2 shows, provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）To BuCh
Esterase has remarkable inhibiting activity, its IC50It is 3.4 μM；More existing widely used anti-AD medicines donepezil is to BuCh ester
The inhibitory activity of enzyme（IC50It is 4.76 μM）It is high.Also, (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）To butyryl courage
The inhibitory activity of alkali esterase（IC50It is 3.4 μM）It is significantly higher than to acetylcholinesterase（IC50It is 5.6 μM）Inhibitory activity, say
Bright compound provided by the present invention（I）There is 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）The method reported
It is measured, i.e.,：Pretreated Aβ 1-42Storing solution is made into DMSO, 50 μ are diluted to the PBS of pH7.4 using preceding
M；Testing compound DMSO is made into 2.5mM storing solutions, and respective concentration is diluted to the PBS of pH7.4 using preceding, takes 20
The A of μ Lβ 1-42The testing compound solution of the μ of solution+20 L, 20 μ L Aβ 1-42The μ L of solution+20 PBSs (contain 2%
DMSO), 20 μ L PBSs (containing 2% DMSO)+20 μ L PBSs (containing 25% DMSO) is changed in the orifice plate of black 96
Compound and Aβ 1-42Ultimate density be 25 μM.37 °C of 24 h of incubation, are subsequently adding 160 μ L and contain 5 μM the 50 of thioflavine T
The glycine-NaOH buffer (pH=8.5) of mM, Varioskan Flash Multimode Reader are used after shaking 5s immediately
Multi-function microplate reader determines fluorescent value under 446 nm excitation wavelengths and 490 nm launch wavelengths；Aβ 1-42+ testing compound it is glimmering
Light value is recorded as IFi, Aβ 1-42The fluorescent value of+PBS is recorded as IFc, the fluorescent value for comprising only PBS is recorded as
IF0, A is suppressed by compoundβ 1-42The inhibiting rate computing formula of self assemble is：100-(IFi-IF0)/(IFc-IF0)×100；Often
Individual compound two multiple holes of each concentration mensuration；With curcumin as positive control.
Experimental result refers to table 3.
Table 3 (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）To Aβ 1-42The inhibitory activity test of self assemble
,
a 25 μM of inhibitor concentration, and the mean of Inhibition was determined at
± SD of the 3 independent experiments.
The result of table 3 shows compound (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）With preferably suppression A β1-42
Aggregation activity, inhibiting rate is 61.1%；The inhibiting rate of positive drug curcumin is 56.8%；Widely used anti-AD medicines donepezil exists
To A β under 25 μM of concentration1-42The inhibiting rate of self assemble is less than 5%.
（3）Antioxidative Activity Determination（ORAC-FL methods）
Reagent and instrument：6- hydroxyl -2,5,7,8- tetramethyl primary colours alkane -2- carboxylic acids（Trolox, purchased from uncommon love (Shanghai) the chemical conversion work of ladder
Industry Development Co., Ltd）With the PBS of 75mM（pH7.4）It is made into the 10-80 μm of solution of ol/L；Fluorescein
（Fluorescein, purchased from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder）With the PBS of 75 mM（pH7.4）Match somebody with somebody
Into the solution of 250 nmol/L；2,2 '-azo diisobutyl amidine dihydrochloride（AAPH, has purchased from splendid remote chemistry science and technology (Shanghai)
Limit company）Using the preceding PBS with 75 mM（pH7.4）It is made into the solution of 40 mmol/L；
ELIASA is Varioskan Flash Multimode Reader（Thermo Scientific）.
Determination experiment method：To addition 50 or 10 μm of μ L of the compound solution of ol/L 20 and fluorescein in the orifice plate of black 96
The μ L of solution 120, mix, 37 °C of 15 min of incubation, add the μ L of AAPH solution 60, make every hole cumulative volume for 200 μ L, mix,
It is immediately placed in Varioskan Flash Multimode Reader instruments, in 485 nm excitation wavelengths and 535 nm transmitted waves
Measure first order fluorescence value per minute under length, the min of METHOD FOR CONTINUOUS DETERMINATION 90 calculates area under fluorescence decay curve by instrument automatically
AUC.Wherein using the 1-8 μm of trolox of ol/L as standard, to be not added with testing sample as blank.The antioxidation activity of compound
Results expression is the equivalent of trolox, and computing formula is：[(AUC Sample-AUC blank)/(AUC Trolox-AUC
blank)][(concentration of Trolox/concentration of sample)].Each compound is determined every time
3 multiple holes, every group of experiment is independent in triplicate.
Experimental result refers to table 4.
Table 4 (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）Antioxidative Activity Determination
The result of table 4 shows, provided by the present invention (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）Anti-oxidant work
Property for 1.1 times of Trolox, with preferable antioxidation activity.
（4）CTA of the compound (I) to PC12 cells
DMEM nutrient solution of the PC12 cells containing 10% hyclone, is 4 × 10 with density4Individual/ml is inoculated in 96 well culture plates
On, inoculation volume is 100 μ l/ holes, is subsequently placed into containing 5%CO237 °C of constant incubators in culture.PC12 cell culture 24 is small
Shi Hou, adds the compound of various concentrations in administration group（I）(final concentration of 100 μM, 10 μM, 1 μM) 10 μ L/ holes, constant temperature training
After supporting 24 h, 3- (4,5- dimethylthiazole -2- bases) -2,5- diphenyltetrazolium bromide bromides (MTT) of 5 mg/mL are added in each group
100 μ L/ holes, carry out living cells dyeing.After the μ L/ holes of 100%DMSO terminate liquids 100 are added after 3 hours, in each group, fully dissolve
Mix.The OD values of each group are determined under the wavelength of 590 nm.Each compound is respectively with 100 μM, 10 μM, 1 μM of test, 3 knots
Really, it is 100% with control group with Duncan ' s test method statistics, administration class value is represented with the percentage of control group.Inhibiting rate
=(control-compound)/control * 100%.
Measurement result refers to Fig. 1, shows compound（I）With relatively low cytotoxicity, possess a treatment model for safety
Enclose.
（5）Compound is to H2O2The protective effect screening of the PC12 cellular damages of induction
DMEM nutrient solution of the PC12 cells containing 10% calf serum, with 1 × 105Individual/mL density is inoculated on 96 well culture plates,
Inoculation volume is 100mL/ holes, is subsequently placed into containing 5%CO237 DEG C of constant incubators in culture.After culture 24 hours, administration group
In plus respective concentration compound（Final concentration of 10 μM, 1 μM）10mL/ holes, preincubate 2 hours（Control group is distinguished with damage group
Plus 10 μ L/ hole PBS, its volume is kept equal）.After PC12 cell incubations 2 hours, it is separately added into administration group with damage group
100μΜH2O2Damage the μ L/ holes of agent 10（Control group adds 10 μ L/ hole PBS）, after 30 minutes, the nutrient solution of each group is changed into without small
The nutrient solution of cow's serum continues to be put into culture 24 hours in constant incubator, and nutrient solution volume thinks 100 μ L/ holes.Continue to cultivate
After 24 hours, each group adds 5mg/mL, and the μ L/ holes of MTT 100 carry out living cells dyeing.After after 3 hours, 100% is added in each group
The μ L/ holes of DMSO terminate liquids 100, fully dissolving mix.The OD values of each group are determined under the wavelength of 490 nm, test result repeats 3
Secondary, with Duncan ' s test method statistics, each group numerical value is expressed as mean ± S.E.M., is 100% with control group, administration group
And damage class value is represented with the percentage of control group.
Measurement result refers to Fig. 2, Fig. 2 show compound (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）It is right
H2O2The PC12 cellular damages of induction have significant neuroprotection.
（6）Compound（I）Acute toxicity test
Test material：Experimental animal is SPF grades of Kunming mouse, is provided by Sichuan Provincial Academy of Traditional Chinese Medicine, produces quality certification number
SCXK (river) 2008-19.Animal feeding is in Sichuan Provincial Academy of Traditional Chinese Medicine pharmacological toxicology research institute SPF barrier systems.It is indoor
20~22 DEG C of temperature, relative humidity 40%-70% or so is illuminated 12 hours and become clear, 12 hours dark, free water.Full nutrition
Grain feed is provided by Sichuan Provincial Academy of Traditional Chinese Medicine Experimental Animal Center.
Experimental technique：Animal is grouped at random：The SPF grades of mouse of 18 ~ 22g 40 is taken, male and female half and half, adaptability is fed two days
Afterwards, 4 groups are randomly divided into by body weight.After water 15h is can't help in fasting, difference gavage compound（I）1000 mg/kg、500 mg/kg、
250 mg/kg, 100 mg/kg, tetra- dosage groups, take administered volume for 0.4 mL/10g, and each group is administered once, and see within continuous 14 days
The death condition of each animal is examined and recorded, is analyzed using Bliss statistical softwares.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 grades of Kunming mouse through (N-1,2,3,4-
Tetrahydro isoquinolyl)-asafoetide acid amides（I）After treatment, there is not anxious poison and the death rate, do not occur hair yet and hold up, be slow in action, close
The phenomenon such as eye and breathing acceleration, it is nontoxic to indicate 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；18-22g
Kunming mouse be purchased from Sichuan scientific tcm institute Experimental Animal Center（Quality certification number：SCXK-Sichuan 2008-19）；Animal
Raise in Sichuan Provincial Academy of Traditional Chinese Medicine pharmacological toxicology research institute SPF barrier systems.H illumination/12 h of feeding environment 12 is dark
Alternately, environment temperature is controlled in 20-22 °C, and humid control is in 50-60%.Full nutritious particle feed is by Sichuan Province's scientific tcm
Institute's Experimental Animal Center is provided.Mouse diving tower instrument（Model ZXC-5Q）Produced by Shandong Academy of Medical Sciences's maintenance of equipment supply station
Experimental technique：60 mouse, 18 ~ 22 g, male and female half and half are randomly divided into 6 groups, i.e. blank control group, model by body weight
Control group, donepezil group（5.0mg·kg-1）, compound（I）High dose group（10.0 mg·kg-1）, compound（I）Middle dose
Amount group（5.0mg·kg-1）, compound（I）Low dose group（2.5mg·kg-1）.Every group of mouse is given by dosage point morning and afternoon
Medicine, successive administration 3 times, 50 min carry out modeling after last dose, other each group intraperitoneal injection east Liang in addition to blank control group
The mgkg of henbane alkali 3-1, successive administration 24 days.20 min carry out step dow n test training after modeling, and animal is put into reaction chamber endoadaptation 3
Min, passes to 36 V alternating currents immediately after, trains 5 min, and record the number of times that each mouse is subject to shock by electricity（Errors number）,
And thus as school grade.Tested after 24 h, every min of mouse assay 5, record is subject to the number of animals that shocks by electricity and the
Once jump off platform incubation period and 5 min in errors number, as a result carry out statistical analysis, all data use average
± standard error（Stand error, S.E.）Represent.Using SPSS11.5 software analysis, the neat selection single factor test variance of variance
Analysis（One-way ANOVA）.Measurement data compares and uses one-way analysis of variance, and the comparing of each group mean is checked using t, knot
Fruit sees Fig. 3 respectively.
Refering to Fig. 3, test result indicate that：The present invention (N- 1,2,3,4- tetrahydro isoquinolyls)-asafoetide acid amides（I）To eastern Liang
Henbane alkali induced mice memory representational role obstacle is respectively provided with the effect of being obviously improved.
Finally illustrate, the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, this area is common
Other modifications or equivalent that technical staff is made to technical scheme, without departing from technical solution of the present invention
Spirit and scope, all should cover in the middle of scope of the presently claimed invention.
Claims (8)
1. (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, it is characterised in that：Chemical constitution is shown in formula I：
。
2. a kind of preparation method of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound, it is characterised in that including with
Lower step：
,
Forulic acid（1）With 1,2,3,4- tetrahydroisoquinolines（2）There is condensation reaction under solvent and condensing agent existence condition, obtain
(N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound（I）.
3. the preparation method of (N-1,2,3,4- tetrahydro isoquinolyls)-asafoetide amide compound as claimed in claim 2, it is special
Levy and be：The solvent be dichloromethane, tetrahydrofuran, toluene in one or more, the condensing agent be EDCI, HOBT,
One or more in DCC, DMAP and Ka Te condensation reagent.
4. (N-1,2,3,4- the tetrahydro isoquinolyls)-asafoetide amide compound or its medicine as described in any one of claim 1 ~ 3
Application of the acceptable hydrate in treatment nerve degenerative diseases medicine is prepared on.
5. a kind of pharmaceutical composition, it is characterised in that：(N-1,2,3,4- tetra- as claimed in claim 1 or 2 including effective dose
Hydrogen isoquinoline base)-asafoetide amide compound or its pharmaceutically acceptable hydrate.
6. pharmaceutical composition according to claim 5, it is characterised in that：Its formulation is oral quick disintegrating tablet, oral cavity compound system
Agent, oral sustained-release preparation, depot long-acting injection or Percutaneously administrable preparation.
7. application of the pharmaceutical composition as claimed in claim 5 in treatment Alzheimer's medicine is prepared.
8. application of the pharmaceutical composition as claimed in claim 6 in treatment Alzheimer's medicine is prepared.
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| CN108276332A (en) * | 2018-01-29 | 2018-07-13 | 南阳师范学院 | A kind of double twin pharmaceutical compounds of asafoetide amide and its preparation method and application |
| CN113336730A (en) * | 2021-06-29 | 2021-09-03 | 山东第一医科大学(山东省医学科学院) | Preparation method and application of 4, 6-dihydroxy-2-phenylbenzofuran |
| CN116102546A (en) * | 2022-11-28 | 2023-05-12 | 浙江海洋大学 | Butyrylcholine esterase and monoamine oxidase B dual inhibitor and application thereof |
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| CN116102546A (en) * | 2022-11-28 | 2023-05-12 | 浙江海洋大学 | Butyrylcholine esterase and monoamine oxidase B dual inhibitor and application thereof |
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