CN1709508A - Use of acetylcholinesterase inhibitor for preparing diabetes medicines - Google Patents
Use of acetylcholinesterase inhibitor for preparing diabetes medicines Download PDFInfo
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- CN1709508A CN1709508A CN 200510027127 CN200510027127A CN1709508A CN 1709508 A CN1709508 A CN 1709508A CN 200510027127 CN200510027127 CN 200510027127 CN 200510027127 A CN200510027127 A CN 200510027127A CN 1709508 A CN1709508 A CN 1709508A
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Abstract
The present invention relates to an application of acetylcholinesterase inhibitor for curing diabetes. The described inhibitor includes all the compounds which can inhibit acetylcholinesterase activity and have the action for curing diabetes. The invented experimental results show that these inhibitors have the actions of inhibiting beta-cell death and reducing blood sugar concentration of diabetic model mouse, and can effectively cure or improve diabetes symptom.
Description
Technical field
The present invention relates to biomedicine field, relate to the application of acetylcholinesteraseinhibitors inhibitors in preparation treatment diabetes medicament particularly.
Background technology
Diabetes are one group of common metabolism endocrinopathys, lack for insulin is absolute or the chronic disease of insulin biological effect due to reducing, main performance blood glucose increases, loses weight, easily accompanying infection (as pulmonary tuberculosis etc.), arteriosclerosis, neuropathy etc., and World Health Organization (WHO) classifies diabetes one of as three big difficult disease.There is diabetics 1.25 hundred million in the whole world, and the diabetics of China has reached 4,000 ten thousand.Prevalence increases with the raising of people's living standard.13 regional total prevalence rates are 0.67% in China, and the big city is about about 1%, and prevalence significantly raises after 40 years old.American-European countries's prevalence reaches 3～6%.Its chronic vascular complication has become one of major reason of crowd's death.
According to the standard of World Health Organization (WHO) in 1999, diabetes mainly are divided into following four types: type i diabetes, and the past claims insulin dependent diabetes mellitus (IDDM) again; Type ii diabetes, the past claims non-insulin-dependent diabetes mellitus again; Gestational diabetes; The Idiotype diabetes.
Wherein present type i diabetes patient can only depend on the exogenous insulin that gives and treat, but insulin injection not exclusively is physiological treatment also, at first enter liver by portal vein unlike interior living insulin, its dosage is artificial the grasp, can not accurately be fit to physiological variation and needs.Medical islets of langerhans have the insulin of pig and cattle, its aminoacid ingredient is not identical entirely with insulin human, and insulin preparation also contains impurity, and allergy (erythra etc.) can take place after the clinical use, injection site adipose hyperplasia, atrophy produce side effect such as insulin antibody and Drug resistance.
The medicine of treatment type ii diabetes mainly contains two big classes at present: a class is that medicine does not directly stimulate insulin secretion, but can improve insulin sensitivity, it is euglycemic agent, this class medicine comprises: biguanides, alpha-glucosidase inhibitor, thiazolidinediones, main effect is to hinder the intestinal absorption nutrient substance, reduce gluconeogenesis function of liver, impel insulin and receptors bind.Another kind of is that medicine can directly stimulate insulin secretion, it is the insulin succagoga, this class medicine comprises: sulfonylurea, as tolbutamide (tolbutamide, tolbutamide, D860), Mellinese (chlorpropamide), glyburide etc., it mainly acts on is to promote the B cells of pancreas excreting insulin, and the increase insulin combines with receptor.
But the problem that biguanides exists has gastrointestinal side effect and lactic acidosis.That gastrointestinal side effect shows as is nauseating, xerostomia, abdominal distention and diarrhoea.Metformin accumulated during lactic acidosis was meant treatment, suppressed the liver aerobic metabolism, caused that lactic acid increases, and lactic acid gathers and causes acidosis in the blood.The patient shows as weak myalgia, dyspnea, symptoms such as confused " deep sound sleep ", hypotension and intractable bradycardia, and case fatality rate is up to 50%.Same sulfonylurea medicine also has certain limitation, mainly shows: 1, treatment mortality height, and the type ii diabetes patient of the new diagnosis of 10-15% is invalid to sulfonylureas, and patient's blood glucose of 30-35% is controlled to some extent, but still belongs to the hyperglycemia level.Among the effective patient of initial therapy, still have every year the patient of 5-10% that this medicine is lost reaction; 2, cause weight increase, the patient of most of build obesities can not use; 3, stimulate insulin secretion and can cause hypoglycemia, insulin level is too high in the blood, may cause cardiovascular complication; 4, through liver metabolism, through kidney and bile excretion, the careful usefulness of the patient of liver, renal insufficiency, serious renal failure and gravid woman's forbidding.
In sum, this area presses for the medicine that diabetes can be effectively treated in exploitation.
Summary of the invention
The invention provides the application in the acetylcholinesteraseinhibitors inhibitors preparation treatment diabetes medicament.
Acetylcholinesteraseinhibitors inhibitors involved in the present invention is meant the chemical compound of a class acetylcholine esterase inhibition activity, and representational inhibitor comprises (but being not limited to): 1, and 5-two [4-allyl dimethyl base ammonium phenyl penta-3-ketone], decamethonium, N, N, N-trimethyl-ortho-aminophenol, gallamine, 3,8-diaminourea-5-[3-(diethylmethyl ammonium) propyl group]-the 6-phenylphenanthridineand, (±)-2-[(1-benzyl-4-piperidyl) methyl]-5,6-dimethoxy-1-Indanone, 3-[1-(benzyl)-4-piperidyl]-1-(2,3,4,5-tetrahydrochysene-1H-1-benzazepine-8-yl)-and 1-acetone, irinotecan, (S)-and N-ethyl-N-methyl-3-[(1-dimethyl-amino) ethyl] carbanilate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol methyl carbamate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-carbanilate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-8-benzyl-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-2 '-methyl phenyl carbamate, (-)-(3aS)-3a, 8-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, (-)-(3aS)-1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, (-)-(3aS)-8-benzyl-1,3a-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, (-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, (-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, (-)-(3aS)-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, ±-uncle 5-[2-fourth amino-1-hydroxyethyl]-position-penylene two-dimethylcarbamate, N, N, N-trimethyl-3-[(dimethylamino) formyloxy] aniline, ambenonium chloride, 1-methyl-3-pyridone father-in-law dimethyl carbamate, 1-naphthyl methyl carbamate, 2-isopropoxy benzene-N-methyl carbamate, 2,3-dihydro-2,2-dimethyl benzofuran-7-phenol methyl carbamate, adjacent cumenyl methyl carbamate, between toluene-N-methyl carbamate, benzimidazolyl-2 radicals-aminocarbamic acid methyl ester, 9-amino-1,2,3, the 4-tetrahydro acridine, 7-methoxyl group tacrine, 11-methyl-3-methoxyl group-4 α, 5,9,10,11,12-six hydrogen-6H-benzofuran [3 α, 2-ef] [2] benzo-aza-6-alcohol, (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also, 7-methyl-5,6,9,10,11,12,13,14-octahydro-5,11-tetrahydropyridine-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also, 12-amino-3-chloro-9-ethyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline, 12-amino-3-chloro-9-methyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline, N-[2-hydroxyl-3-methoxyl group-5-chloro-benzal]-huperzine A, 12-(2 '-carboxyl-5 '-anisyl)-2,12-dihydroxy-dodecane-4-ketone, O, the O-DFP, N-(phosphoric acid methyl) glycine, O, O-dimethyl-(2,2,2-three chloro-1-hydroxyethyls) phosphate ester, O, O-dimethyl-O-(2, the 2-dichloro) vinyl phosphate, O, O-dimethyl-O-p-nitrophenyl thiophosphate, O, O-dimethyl-S-(N-methylamino formyl methyl) phosphorodithioate, O, O-dimethyl-S-(N-methylamino formyl methyl) thiophosphate, O, O-dimethyl-S-[2-(1, the 4-diethyl succinate) base] phosphorodithioate, Phenylmethanesulfonyl fluoride, methanesulfonyl fluoride, the tetra isopropyl pyrophosphoramide, Fasciculin-I, Fasciculin-II and their derivant etc.Enumerate compound structure referring to Fig. 1.These chemical compounds all are known, can the synthetic or purchase from the market according to conventional method (as reported method in document and the patent).
Wherein preferred inhibitors be (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone (huperzine A, huperzine A) also
Acetylcholinesteraseinhibitors inhibitors can with by pharmaceutically or the deutero-salt form of the acceptable acid of physiology or alkali use.These salt include, but is not limited to the salt with following mineral acid formation: example hydrochloric acid, sulphuric acid, nitric acid, phosphoric acid and the salt that forms with organic acid, organic acid then refers to acetic acid, oxalic acid, succinic acid, tartaric acid, methanesulfonic acid and maleic acid.Other salt comprise the salt that forms with alkali metal or alkaline-earth metal (as sodium, potassium, calcium or magnesium), with the form (when with this form administration, can change into active part in vivo) of " prodrug " of ester, carbamate or other routines.
Diabetes involved in the present invention comprise type i diabetes and type ii diabetes, wherein are preferably type i diabetes.
The ultimate principle opinion of institute of the present invention foundation: diabetes are one group of heterogeneous metabolic diseases, and its pathogenesis is very complicated, and apoptosis is the another focus of noting in recent years.Bronwyn etc. (O ' Brien BA, et al.Diabetes, 1997,46:750-757.) discover that apoptotic cells is a beta cell in the islets of langerhans.In type i diabetes, think at present owing to reduce at beta Cell of islet autoreactivity T, bone-marrow-derived lymphocyte apoptosis in the body, make beta cell be subjected to its attack and many (the Eisenbarth GS.N Engl J Med of increasing apoptosis take place, 1986,21:1360-1368.), the cytokine of immunologically competent cell generation such as IL-1, TNF and alkylating agent or NO also mediate beta cell apoptosis (Stassi G in addition, Todaro M, et al.Diabetes, 1996,45 (Suppl): 240A.).And the pancreas amyloid polypeptide of type ii diabetes patient pancreatic can induce pancreatic generation apoptosis (Ankarcrona M, et al.Exp Cell Res, 1994,213:172-177.).In the generating process of diabetes, because the pancreatic apoptosis increases, its function is reduced, may be to cause one of pathogenetic mechanism of glycosuria.
Acetylcholinesterase (Acetylcholinesterase, AChE) except having hydrolysis acetylcholine apace, stop outside the function of cholinergic nerve signal transmission, in the increment of differentiation, migration, synapse formation and the hematopoietic cell and the tumor cell of neurocyte and differentiation regulation process, has important function (Small, DH., Neurochem Int, 1996,28:453-483).Recent findings AChE also participate in the neuronal apoptosis process (Yang Lei et al., Neuroscience Res, 2002,42:261-268).
Though the apoptosis of beta cell plays significant feature in the diabetes process, never have the report that AChE works in the beta cell apoptotic process.The present invention confirms not only diabetes model in vivo but also can both find to have the expression of AChE in external diabetes model in the apoptosis of beta cell, do not find the activity of AChE in the normal pancreas section, but along with increasing the weight of of the diabetes state of an illness, the activity of increasing AChE can appear.The present invention simultaneously adopts acetylcholinesteraseinhibitors inhibitors to treat diabetic mice effectively, improves the symptom of its hyperglycemia.
In the present invention, use as giving a definition:
Herein, term " contain " expression various compositions can be applied to pharmaceutical composition of the present invention together.Therefore, term " mainly by ... form " and " by ... composition " be included in during term " contains ".
Herein, " pharmaceutically acceptable " composition is to be applicable to people and/or animal and not have excessive bad side reaction (as toxicity, stimulation and allergy), and the material of rational benefit/risk ratio is promptly arranged.More preferably, " pharmaceutically can be accepted " and be meant and do not influence the active innocuous substance of maincenter acetylcholinesteraseinhibitors inhibitors.
Be enough to obtain the therapeutic response of needs when herein, term " safe and effective amount " refers to use by mode of the present invention and do not have the amount that excessive bad side reaction (as toxicity, stimulation and allergy) has the composition of rational benefit/risk ratio.Obviously, concrete " safe and effective amount " is different because of various factors, as the structure of kind (if any), applied concrete preparation and the chemical compound or derivatives thereof of the subject special state of an illness, patient's body condition, subject mammiferous kind, the course of treatment, the treatment carried out simultaneously.
Herein, " active component " comprises acetylcholinesteraseinhibitors inhibitors and pharmaceutically acceptable salt thereof.Representational salt comprises (but being not limited to): sodium salt, potassium salt, zinc salt etc.
In this article, term " acetylcholinesteraseinhibitors inhibitors " comprises acetylcholinesteraseinhibitors inhibitors and salt thereof.
Herein, " pharmaceutical carrier " is pharmaceutically acceptable solvent, suspending agent or the excipient that is used for acetylcholinesteraseinhibitors inhibitors is sent to the animal or human.Carrier can be a liquid or solid, can select according to administering mode.
Reactive compound
The reactive compound that the present invention is used for the treatment of diabetes is an acetylcholinesteraseinhibitors inhibitors.Acetylcholinesteraseinhibitors inhibitors is the chemical compound of a class acetylcholine esterase inhibition activity, representational inhibitor comprises (but being not limited to): methyl (±)-2-[(1-benzyl-4-piperidyl)]-5,6-dimethoxy-1-Indanone, 3-[1-(benzyl)-4-piperidyl]-1-(2,3,4,5-tetrahydrochysene-1H-1-benzazepine-8-yl)-1-acetone, (S)-and N-ethyl-N-methyl-3-[(1-dimethyl-amino) ethyl] carbanilate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol methyl carbamate, 9-amino-1,2,3, the 4-tetrahydro acridine, 11-methyl-3-methoxyl group-4 α, 5,9,10,11,12-six hydrogen-6H-benzofuran [3 α, 2-ef] [2] benzo-aza-6-alcohol, (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also, O, O-dimethyl-(2,2,2-three chloro-1-hydroxyethyls) phosphate ester, the tetra isopropyl pyrophosphoramide, Fasciculin-I and Fasciculin-II and their derivant etc.Enumerate compound structure referring to Fig. 1.These chemical compounds all are known, can the synthetic or purchase from the market according to conventional method (as reported method in document and the patent).
A kind of particularly preferred inhibitor be (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone (huperzine A, huperzine A) also
Acetylcholinesteraseinhibitors inhibitors can with by pharmaceutically or the deutero-salt form of the acceptable acid of physiology or alkali use.These salt include, but is not limited to the salt with following mineral acid formation: example hydrochloric acid, sulphuric acid, nitric acid, phosphoric acid and the salt that forms with organic acid, organic acid then refers to acetic acid, oxalic acid, succinic acid, tartaric acid, methanesulfonic acid and maleic acid.Other salt comprise the salt that forms with alkali metal or alkaline-earth metal (as sodium, potassium, calcium or magnesium), with the form (when with this form administration, can change into active part in vivo) of " prodrug " of ester, carbamate or other routines.
Dosage
In the present invention, the dosage to acetylcholinesteraseinhibitors inhibitors is not particularly limited available any proper dosage.Diabetes comprise constitutional and Secondary cases.When using acetylcholinesteraseinhibitors inhibitors treatment diabetes, amount of drug depends on the situation etc. of character, the course of disease, weight, drug reaction situation and the patient undergoing treatment of disease, finally give patient how much dosage by prescription doctor decision, generally can be with reference to the consumption that is applied to AD patient.
Generally, suitable content is the 0.01%-99% that acetylcholinesteraseinhibitors inhibitors accounts for the pharmaceutical composition gross weight, preferably 0.1-90%.When the treatment diabetes, the effective dosage ranges of acetylcholinesteraseinhibitors inhibitors is generally 0.005-50 mg/kg sky or higher, preferably is 0.01-5 mg/kg sky, more preferably is 0.05-1 mg/kg sky.
Dosage device comprises a kind of acetylcholinesteraseinhibitors inhibitors chemical compound, the formed mixture of perhaps multiple acetylcholinesteraseinhibitors inhibitors chemical compound.Dosage device also can contain diluent, filler, carrier etc.Dosage unit is solid or gel form, as pill, tablet, capsule etc., or liquid form, they are fit to oral administration, rectally, topical or parenteral or intravenous administration.
Dosage form
Peroral administration solid composite of the present invention can adopt forms such as tablet, pill, capsule, powder, granule, drop.Mixed active substance and at least one inert diluent more than a kind or a kind in these solid composites, for example, lactose, mannitol, glucose, hydroxypropyl cellulose, microcrystalline Cellulose, starch, polyvinylpyrrolidone, agar, pectin, aluminosilicate magnesium, magnesium aluminate.Also can make the additive that contains in the compositions except inert diluent according to common method, for example, cosolvents such as stabilizing agents such as disintegrating agents such as lubricants such as magnesium stearate, glycolic cellulose calcium, lactose, glutamic acid or aspartic acid.Tablet or pill in this way also can be as required, sugar-coat such as sucrose, gelatin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose phthalate or gastric solubility, enteric film on its outer wrap.
Peroral administration fluid composition comprises the opacifiers that allows on the medicament, solution, suspending agent, syrup, elixir etc., and normally used inert diluent comprises Purified Water, ethanol.Except inert diluent, also can comprise auxiliary agents such as wetting agent, suspending agent, sweeting agent, correctives, aromatic and antiseptic in the said composition.
Para-oral injection comprises sterile aqueous or non-aqueous solution agent, suspending agent and opacifiers.Comprise injection in aqueous solution agent and the suspending agent with distilled water and normal saline.Comprise propylene glycol in water-insoluble solution and the suspending agent, Polyethylene Glycol, cocoa butter, olive oil, Semen Ricini wet goods vegetable oil, alcohols such as ethanol, arabic gum, Tween 80 (trade name) etc.Also can comprise isotonic agent, antiseptic, wetting agent, emulsifying agent, dispersant, stabilizing agent (for example, lactose), cosolvent (for example, glutamic acid, aspartic acid) in these compositionss.With bacteriological filtration membrane filtration above-mentioned composition, be used biocide again and just can reach aseptic purpose.Then, utilize above-mentioned composition to make aseptic solid composite, water or aseptic injection just can be used with dissolution with solvents before use.
Can be used for preparing pharmaceutically acceptable carrier peroral dosage form of the present invention, concrete and excipient example,, in 297 (JIUYUE was authorized Robert on the 2nd in 1975) description is arranged in U.S. Patent No. 3,903.Be used to make the technology and the compositions of useful dosage form of the present invention, description is arranged in following document: 7 kinds of modern preparations (7ModernPharmaceutics). the 9th and 10 chapter (Banker; Rhodes edits, and 1979); People such as Lieberman, pharmaceutical dosage form: tablet (Pharmaceutical Dosage Forms:Tablets) (1981); And Ansel, pharmaceutical dosage form introduction (Introduction to Pharmaceutical Dosage Forms) 2 editions (1976).
Therapeutic Method
Therapeutic Method can be any suitable effective ways of treatment diabetes.Treatment can be oral administration, rectally, topical, parenteral, intravenous administration.The method of using the effective dose medicine also depends on the symptom and the severity extent of diabetes to be treated.It is believed that by acetylcholinesteraseinhibitors inhibitors is prepared with suitable carriers or diluent, and the parenteral Therapeutic Method by oral, intravenous, subcutaneous or intramuscular administration, be with the method for optimizing of compound administration in homoiothermic animal.
Acetylcholinesteraseinhibitors inhibitors of the present invention also can with the material coupling of other treatment and adjuvant therapy of diabetes, with further raising therapeutic effect.
Major advantage of the present invention is: acetylcholinesteraseinhibitors inhibitors can reach the purpose that stops the beta cell apoptosis by acetylcholine esterase inhibition very effectively, thereby has protected the beta cell of diabetic, has improved the function of islets of langerhans.Therefore, acetylcholinesteraseinhibitors inhibitors can be effectively to the treating of diabetes, the diabetics to the I type more can improve curative effect especially.
Description of drawings
The chemical structural drawing of Fig. 1 acetylcholinesteraseinhibitors inhibitors chemical compound.
1:1,5-two (4-allyl dimethyl base ammonium phenyl) penta-3-ketone
1，5-bis(4-allyldimethylammoniumphenyl]-pentan-3-one，BW284c51，C
25H
38N
2O
2: decamethonium (decamethonium, C16H38N2)
3:N, N, N-trimethyl-ortho-aminophenol
N，N，N-trimethyl-O-hydroxyaniline，Edrophone，C
9H
14NO
4: gallamine (gallamine, C
30H
60N
3O
3)
5:3,8-diaminourea-5-[3-(diethylmethyl ammonium) propyl group]-the 6-phenylphenanthridineand
3，8-Diamino-5-[3-(diethylmethylammonio)propyl]-6-phenyl?phenanthridiniumPropidium，C
27H
34N
4
6:(±)-2-[(1-benzyl-4-piperidyl) methyl]-5,6-dimethoxy-1-Indanone
(±)-2-[(1-benzylpiperidin-4-yl)methyl]-5，6-dimethoxy-indan-1-one，donepezil，C
24H
29NO
3
7:3-[1-(benzyl)-4-piperidyl]-1-(2,3,4,5-tetrahydrochysene-1H-1-benzazepine-8-yl)-1-acetone
3-[1-(Phenylmethyl)-4-Piperidinyl]-1-(2，3，4，5-Tetrahydro-1H-1-Benzazepin-8-yl)-1-Propanone，TAK-147，C
25H
32N
2O
8: irinotecan (Irinotecan, CPT11, C
33H
39N
4O
6)
9:(S)-and N-ethyl-N-methyl-3-[(1-dimethyl-amino) ethyl] carbanilate
(S)-N-ethyl-N-methyl-3-[(1-dimethyl-amino)ethyl]phenylcarbamate，rivastigmine，C
14H
22N
2O
2
10:1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol methyl carbamate
1，2，3，3a，8，8a-hexahydro-1，3a，8-trimethylpyrrolo[2，3-b]indol-5-ol?methylcarbamate，Physostigmine，C
15H
21N
3O
2
11:1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-carbanilate
1，2，3，3a，8，8a-hexahydro-1，3a，8-trimethylpyrrolo[2，3-b]indol-5-ol-N-phenylcarbamate，Phenserine，C
20H
23N
3O
2
12:1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate
1，2，3，3a，8，8a-hexahydro-1，3a，8-trimethylpyrrolo[2，3-b]indol-5-yl-N-4′-isopropylphenylcarbamate，Cymserine，C
23H
29N
3O
2
13:(-)-(3aS)-and 3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-3a，8-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-4′-Isopropylphenylcarbamate，N1-norcymserine，C
22H
27N
3O
2
14:(-)-(3aS)-and 1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-1-phenethyl-3a，8-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-4′-Isopropylphenylcarbamate，N1-phenethylnorcymserine，C
30H
35N
3O
2
15:(-)-(3aS)-and 8-benzyl-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-8-Benzyl-1，3a-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl N-4′-Isopropylphenylcarbamate，C
29H
33N
3O
2
16:(-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-1，3a-Dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo-[2，3-b]indol-5-yl?N-4′-Isopropylphenylcarbamate，C
22H
27N
3O
2
17:(-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-1，8-Dibenzyl-3a-methyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-4′-Isopropylphenylcarbamate，C
35H
37N
3O
2
18:(-)-(3aS)-and the 3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also
(-)-(3aS)-3a-methyl-1，2，3，3a，8，8a-hexahydropyrrolo-[2，3-b]indol-5-yl?N-4′-Isopropylphenylcarbamate，C
21H
25N
3O
2
19:1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-2 '-methyl phenyl carbamate
1，2，3，3a，8，8a-hexahydro-1，3a，8-trimethylpyrrolo[2，3-b]indol-5-ol-N-2′-Methylphenylearbamate，Tolserine，C
21H
25N
3O
2
20:(-)-(3aS)-and 3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-3a，8-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-2′-methylphenylcarbamate，N1-nortolserine，C
20H
23N
3O
2
21:(-)-(3aS)-and 1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-1-phenethyl-3a，8-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-2′-methylphenylcarbamate，N1-phenethylnortolserine，C
28H
30N
3O
2
22:(-)-(3aS)-and 8-benzyl-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-8-Benzyl-1，3a-dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-2′-Methylphenylcarbamate，C
27H
29N
3O
2
23:(-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-1，3a-Dimethyl-1，2，3，3a，8，8a-hexahydropyrrolo-[2，3-b]indol-5-yl?N-2′-Methylphenylcarbamate，C
20H
23N
3O
2
24:(-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-1，8-Dibenzyl-3a-methyl-1，2，3，3a，8，8a-hexahydropyrrolo[2，3-b]indol-5-yl?N-2′-Methylphenylcarbamate，C
33H
33N
3O
2
25:(-)-(3aS)-and the 3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also
(-)-(3aS)-3a-methyl-1，2，3，3a，8，8a-hexahydropyrrolo-?[2，3-b]indol-5-yl?N-2′-Methylphenylcarbamate，C
19H
21N
3O
2
26: ±-uncle's 5-[2-fourth amino-1-hydroxyethyl]-position-penylene two-dimethylcarbamate
±-5-[2-(tert-butylamino)-1-hydroxyethyl]-m-phenylene?bis-dimethylcarbamate，bambuterol，C
18H
29N
3O
5
27:N, N, N-trimethyl-3-[(dimethylamino) formyloxy] aniline N, N, N-trimethyl-3-[(dimethyl-amino) formyloxy] aniline, neostogmine, C
12H
19N
2O
2
28: ambenonium chloride (Ambenonium, C
29H
46Cl
2N
4O
2)
29:1-methyl-3-pyridone father-in-law dimethyl carbamate
1-methyl-3-hydroxy-pyridinium?dimethylcarbamate，Pyridostigmine，C
9H
13N
2O
2
30:1-naphthyl methyl carbamate
1-naphthyl?methylcarbamate，sevin，carbaryl，C
12H
11NO
2
31:2-isopropoxy benzene-N-methyl carbamate
2-isopropoxyphenyl-N-methylcarbamate，Propoxur，C
11H
15NO
3
32:2,3-dihydro-2,2-dimethyl benzofuran-7-phenol methyl carbamate 2,3-dihydro-2,2-dimethylbenzofuranol-7-ol methylcarbamate, C
12H
15NO
3
33: adjacent cumenyl methyl carbamate
O-cumenyl?methylcarbamate，isoprocarb，C
11H
15NO
2
34: a toluene-N-methyl carbamate
meta-tolyl-N-methylcarbamate，C
9H
10NO
2
35: benzimidazolyl-2 radicals-aminocarbamic acid methyl ester
benzimidazole-2-yl-amino?formic?acid?methyl?ester，C
9H
9N
3O
2
36:9-amino-1,2,3, the 4-tetrahydro acridine
9-amino-1，2，3，4-tetrahydroacridine，Tacrine，C
13H
14N
2
37:7-methoxyl group tacrine (7-Methoxytacrine, C
14H
16N
2O)
38:11-methyl-3-methoxyl group-4 α, 5,9,10,11,12-six hydrogen-6H-benzofuran [3 α, 2-ef] [2] benzo-aza-6-alcohol
11-methyl-3-methoxy-4α，5，9，10，11，12-hexahydro-6H-benzofuran[3α，2-ef][2]benzoaza-6-ol，Galanthamine，C
17H
21NO
3
39:(5R, 9R, 11E)-and 5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone or huperzine A also
5R，9R，11E-5-amino-11-ethylidene-5，6，9，10-tetrahydro-7-methyl-5，9-methylenecycloocta-(b)pyridine-2(1H)-one，Huperzine?A，C
15H
18N
2O
40:7-methyl-5,6,9,10,11,12,13,14-octahydro-5,11-tetrahydropyridine-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also
7-methyl-5，6，9，10，11，12，13，14-octahydro-5，11-tetrahydropyridine-5，9-methylenecycloocta-(b)pyridine-2(1H)-one，Huperzine?B，C
17H
18N
2O
41:12-amino-3-chloro-9-ethyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline
12-Amino-3-chloro-9-ethyl-6，7，10，11-tetrahydro-7，11-methanocycloocta[b]quinoline，Huperzine?X，C
18H
19ClN
2)
42:12-amino-3-chloro-9-methyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline
12-Amino-3-chloro-9-methyl-6，7，10，11-tetrahydro-7，11-methanocycloocta[b]quinolineHuperzine?Y，C
17H
17ClN
2)
43:N-[2-hydroxyl-3-methoxyl group-5-chloro-benzal]-huperzine A
N-[2-hydroxy-3-methoxy-5-chloro-benzilidene]-huperzine?A，ZT-1，C
23H
23ClN
2O
3
44:12-(2 '-carboxyl-5 '-anisyl)-2,12-dihydroxy-dodecane-4-ketone
12-(2’-carboxy-5’-methoxyphenyl)-2，12-dihydroxy-dodeca-4-one，Sporotricolone，C
20H
31O
6
45:O, the O-DFP
O，O-diisopropyl?fluorophosphate，DFP，Isoflurophate，C
6H
14FO
3P
46:N-(phosphoric acid methyl) glycine
N-(Phosphonomethyl)glycine，glyphosate，C
3H
8NPO
5
47:O, O-dimethyl-(2,2,2-three chloro-1-hydroxyethyls) phosphate ester
O，O-dimethyl-(2，2，2-trichloro-1-hydroxyethyl)phosphate，Metrifonate，Trichlorfon，Totalene，Higalfon，Pronto，C4H8Cl3O4P
48:O, O-dimethyl-O-(2, the 2-dichloro) vinyl phosphate
O，O-dimethyl-O-(2，2-dichlorovinyl)phosphate，DDVP，C
4H
7Cl
2O
4P
49:O, O-dimethyl-O-p-nitrophenyl thiophosphate
O，O-dimethyl-O-paranitrobenzylthiophosphate，Parathion-methyl，Dalf，Folidol-M，C
8H
10NO
5PS
50:O, O-dimethyl-S-(N-methylamino formyl methyl) phosphorodithioate
O，O-dimethyl-S-(N-methylformamylmethyl)dithiophosphate，C
5H
12NO
3PS
2
51:O, O-dimethyl-S-(N-methylamino formyl methyl) thiophosphate,
O，O-dimethyl-S-(N-methylformamylmethyl)thiophosphate，Omethoate，Folemate，C
5H
12NO
4PS
52:O, O-dimethyl-S-[2-(1, the 4-diethyl succinate) base] phosphorodithioate
O，O-dimethyl-S-[2-(1，4-diethyl?succinate)]dithiophosphate
53: Phenylmethanesulfonyl fluoride (phenylmethanesulfonyl fluoride, PMSF, C
7H
7FO
2S)
54: methanesulfonyl fluoride (methanesulfonyl fluoride, MSF, CH
3FO
2S)
55: the tetra isopropyl pyrophosphoramide
tetraisopropylpyrophosphoramide，Iso-OMPA，C
12H
28N
4O
3P
2S
Fig. 2 .RIN-5F apoptosis detects.A:MTT detects and finds that STZ reduces the activity of insulinoma cell strain RIN-5F, and is dose-dependence; B: flow cytometer detects and finds that also apoptosis rate also increases along with the increase of STZ concentration.The apoptosis rate of normal cell nearly 5%, and behind the STZ processing 24h with 10mM, apoptosis rate is 52%; C:DNA ladder electrophoresis.Lane 1: not dosing group, Lane 2:5mM STZ processed group, Lane3:DNA marker, induce RIN-5F cell 24h with the STZ of 5mM after, can find tangible DNA ladder phenomenon.
Fig. 3 .RT-PCR detects AChE expression in the RIN-5F cell under the STZ processing.Lane 1: not dosing group, Lane 2:5mM STZ processed group, Lane 3:10mM STZ processed group.After STZ handled, the RIN-5F cell began to express AchE, did not express AchE and add the RIN-5F cell that STZ handles.
Fig. 4. the detection of AChE in the inductive RIN-5F apoptosis of STZ.A: the bright field video of cell acetylcholinesterase chemical staining, B: with A one visual field fluorescence video, cell is examined with Hoechst 33258 (blueness) transfect cell.Apoptotic cell has obvious characteristics, concentrates such as chromatin, even forms apoptotic body (shown in the arrow).Acetylcholinesterase chemical staining and Hoechst33258 positive mark position overlap.Explanation under apoptotic state, acetylcholinesterase be appear at endonuclear.X400
Fig. 5. the acetylcholinesterase chemical staining of diabetic mice islets of langerhans section.A: the islets of langerhans section acetylcholinesterase chemical staining of the inductive diabetic mice different time points of normal mouse x100:STZ: B: the 8th day islets of langerhans section x100; C: the 15th day islets of langerhans section x200; D: the 22nd day islets of langerhans section x200; E: the 29th day islets of langerhans section x100; Time started to inject for the first time STZ on the 0th day; Pale brown precipitation is represented the positive of acetylcholinesterase in the tissue.
Fig. 6. external acetylcholinesteraseinhibitors inhibitors suppresses the RIN-5F apoptosis.
Fig. 7. acetylcholinesteraseinhibitors inhibitors is to the influence of diabetic mice blood sugar concentration.
Specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The concrete steps of experimental technique commonly used in the following example:
The chemical staining of acetylcholinesterase
After section or the cell fixation, add reactant liquor (15ml 0.1M sodium phosphate pH6.0,10mgacetylthiocholine iodide, 1ml 0.1M sodium citrate solution, 2ml 30mM copper sulphatesolution, 2ml 5mM potassium ferricyanide), room temperature reaction 4-8 hour.After the cyclic washing stopped reaction, tissue slice haematoxylin redyeing 20min, dehydration mounting.Cell reacts 30min, microscopic examination with Hoechst 33258 at 37 ℃.
The extraction of DNA ladder
Behind the STZ processing 24h of RIN-5F cell with 5mM, collecting cell, gene mentation group extracting solution (50mM Tris-cl, PH8.0,100mM EDTA, 1%SDS, 100 μ g/ml E.C. 3.4.21.64s, 1mM NaCl), 37 ℃ of reaction 16h use the conventional extracting of phenol one chloroform, the centrifugal 10min of 12000rpm then.0.7 the different third pure precipitation of volume, behind-70 ℃ of placement 30min, the centrifugal 30min of 12000rpm.Be dissolved among the 20 μ l TE 1.5% sepharose electrophoresis.
Flow cytometer detects apoptosis
5 * 10
5The RIN-5F cell handle after, behind the centrifugal 10min of 200g, the ice ethanol with 70% is 30min fixedly, uses 4 ℃ of lucifuges of propidium iodide (PI) dye liquor (75 μ g/ml PI, 100mg/ml RNase A) to hatch 30min then.Flow cytometer 585nm exciting light detects (FACS calibur, Becton Dickinson) at least 10000 cells.Calculate the ratio of apoptotic cell according to the dna content of cell.
The MTT reaction
The MTT reaction is used to detect cell activity.Cell dilution is inoculated in 96 orifice plates with the 200ul volume to suitable concentration.Cultivate and induce processing after 24 hours.Behind pending the finishing, the MTT that adds 20ul (5mg/ml) again, 37 ℃ 4 hours, last every hole adds 100ul solution (20% sodium dodecyl sulfate in 50%N, N-dimethylformamide, 04%acetic acid, and 0.04N HCl) reaction, after the dissolving to be crystallized fully, detect in 590nm.If the cytoactive height, detected value height then, on the contrary then low.
The experimental technique of other unreceipted actual conditions, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer
Experiment material: STZ (streptozotocin), c2-ceremide, BW284c51, iso-OMPA
(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and eserine are available from Sigma (St.Louis, MO) company for MTT.Two anti-(comprise fluorescence and HRP coupled two anti-) of all kinds is all available from SantaCruz (Santa Cruz, CA) company.ECL reagent is available from Santa Cruz (Santa Cruz, CA) company.Acetylcholinesterase antibody: (BD Biosciences, Franklin Lakes is NJ) with BeiJing, China Military Medical Science Institute available from BD company for monoclonal antibody.Multi-resistance is available from Santa Cruz (Santa Cruz, CA) company.The C57BL/6J mice is from Chinese Academy of Sciences's animal center.
The preparation of embodiment 1 diabetes model
The C57BL/6J mice in 6-8 week, the conventional raising.STZ was dissolved in (PH4.5) among the sodium citrate buffer solution of 0.1M the continuous lumbar injection of 45mg/kg 5 days before injection.Blood glucose monitoring system with Luo Shi biweekly detects blood glucose.Blood sugar concentration is greater than 13.9mM (250mg/ml) time, thinks that diabetes model sets up.
The expression of AChE in the beta cell of apoptosis in the embodiment 2 external diabetes models
1.STZ can induce the RIN-5F apoptosis external
The insulinoma cell strain RIN-5F of rat with 1640 culture medium (15% calf serum, the 1.0mM Sodium Pyruvate, the 4.5g/l glucose, 10mMHEPES) at 37 ℃, 5%CO
2Cultivate.When the cell growth reaches 70% density, use 5mMSTZ, 10mM STZ after 15mM STZ induces 24h, adopts mtt assay, flow cytometer to detect and DNA ladder detection apoptosis.
Result: find that with mtt assay the STZ of various dose can make the activity of insulinoma cell strain RIN-5F reduce (Fig. 2 A).The STZ of 16mM can make the vigor of cell be reduced to 40%.Flow cytometer detects finds that also the ratio of apoptosis also increases along with increasing that STZ measures.The apoptosis rate of normal cell nearly 5%, and behind the STZ processing 24h with 15mM, apoptosis rate can be increased to 52% (Fig. 2 B).After inducing RIN-5F cell 24h with the STZ of 5mM, can find tangible DNA ladder phenomenon (Fig. 2 C).
2. the expression of AChE is arranged in the inductive RIN-5F of STZ
Detect the variation of acetylcholinesterase mRNA level in the RIN-5F of apoptosis cell with the method for RT-PCR.Extract total RNA with TRIzol (GIBCO) from 7 * 106RIN-5F cell, the random primer with Promega carries out reverse transcription then.Then carry out the PCR reaction with the special primer of acetylcholinesterase.What use is PTC-100TM type PCR instrument (MJ.Research INc.).The PCR circulation was made up of following several steps:
94 ℃ of 60see of degeneration (95 ℃ of 5min of the first step),
The 65 ℃ of 60sec that anneal,
Extend 72 ℃ of 60sec (72 ℃ of 10min of final step)
Totally 30 circulate, obtain the band of 440bp.
The primer sequence of AChE is:
forward?primer(1)5’-CCGGAATTCCCGTGCCTCCACATTGACTTG-3’；
Reverse primer (2) 5 '-CCGGCTAGGTGCTATAGTGGTCGAACTGGTTCG-3 '. the primer sequence of β-actin is:
forward?primer(3)5′-AACGAGCGGTTCCGATGCCCTGAG-3′；
reverse?primer(4)5′-TG?TCGCCTTCACCGTTCCAGTT-3′.
After result: STZ handled, the RIN-5F cell began to express AchE, did not express AchE (Fig. 3) and add the RIN-5F cell that STZ handles.
3. the detection of AChE in the inductive RIN-5F apoptosis of STZ
Apoptotic cell has obvious characteristics, concentrates such as chromatin, even forms apoptotic body.The RIN-5F cell is handled with STZ, and the acetylcholinesterase chemical staining is then with Hoechst 33258 single-minded labeled cell nuclears, microscopic examination.
Result: find that acetylcholinesterase chemical staining and Hoechst33258 positive mark position overlap.Explanation is under apoptotic state, and acetylcholinesterase is to appear at endonuclear (Fig. 4).
The expression of AChE in the embodiment 3 diabetic mice beta cells
After putting to death experiment mice, pancreas is with the fixing 24h of 4 ℃ of 4% paraformaldehyde (PFA) solution, then with 30% sucrose solution immersion 12h, and after the usefulness OCT embedding, 10 μ M frozen section, acetylcholinesterase chemical stainings then.
The result: in the inductive diabetes model islets of langerhans section of STZ, beta cell has the active appearance of tangible AChE.The positive staining (Fig. 5 A) that in the pancreas section of normal mouse, does not have AChE.On the contrary, along with the apoptosis and the lymphocytic infiltration (from the 8th day to the 29th day) of beta cell in the diabetes process, can find that the specific staining of AChE appears at around islets (Fig. 5 B-E).Particularly in the time of diabetic symptom severity (from the 22nd day to the 29th day), the destruction of β cell is quite serious, and the dyeing of AChE is strong (Fig. 5 D, E) very.
Embodiment 4. external acetylcholinesteraseinhibitors inhibitors suppress the RIN-5F apoptosis
The RIN-5F cell adds 5mM STZ respectively, 10 μ M BW284c51,5mM STZ+10 μ M BW284c51,10 μ M Tacrine, 5mM STZ+10 μ M Tacrine sets up not dosing group of RIN-5F cell as negative control simultaneously, handled 24 hours, flow cytometer detects apoptosis rate then.
The special inhibitor B W284c51 of result: AChE and Tacrine detect the regulating action of the activity of inhibition AChE for apoptosis.We find that two kinds of inhibitor can reduce the apoptosis of the caused RIN-5F of STZ (Fig. 6).After adding the BW284c51 of 10 μ M, can make apoptosis rate reduce to 10% (P＜0.01), and after adding the Tacrine of 10 μ M, can make apoptosis rate reduce to 17% from 27% of the STZ that only adds 5mM.
Diabetic mice carries out the Drug therapy experiment by following grouping: the STZ group; The STZ+PBS group; The STZ+1mg/kgTacrine group; STZ+0.25mg/Kg Huperzine A group; STZ+0.5mg/Kg Huperzine A group; All medicines are lumbar injection (1 day twice), from STZ injection start injection the previous day, till putting to death.Begin from injectable drug, on an empty stomach behind the 6h, got blood from the mice tail end and survey blood glucose every 4 days.
The result: the blood glucose of STZ+0.25mg/Kg Huperzine A, two groups of STZ+0.5mg/Kg Huperzine A and STZ+1mg/kgTacrine group is along with the prolongation of time progressively descends, especially Huperzine A, in medication after 30 days, the blood glucose of mice has been reduced to the level of normal mouse, and STZ group and STZ+PBS group are all kept hyperglycemia always, and exist difference (P＜0.01) significantly between the administration group.(Fig. 7)
The preparation of embodiment 6 tablets:
Prescription: (1000)
Huperzine A 200 grams
Hydroxypropyl cellulose 700 grams
Glycolic cellulose calcium 100 grams
Preparation method:
With above-mentioned material, adopt the method preparation of this area routine to become tablet, every weight is 1 gram, active ingredient content is 0.2 gram.
Data statistics
With the result of SigmaPlot software analysis FACS, income analysis result represents with means ± SE, the notable difference between Student ' st-test and one-way ANOVA analytical data, the standard P of significant difference＜0.05.
Claims (7)
1. acetylcholinesteraseinhibitors inhibitors is preparing the purposes for the treatment of in the diabetes medicament.
2. purposes as claimed in claim 1 is characterized in that, described inhibitor is selected from down group:
1,5-two [4-allyl dimethyl base ammonium phenyl penta-3-ketone], decamethonium, N, N, N-trimethyl-ortho-aminophenol, gallamine, 3,8-diaminourea-5-[3-(diethylmethyl ammonium) propyl group]-the 6-phenylphenanthridineand, (±)-2-[(1-benzyl-4-piperidyl) methyl]-5,6-dimethoxy-1-Indanone, 3-[1-(benzyl)-4-piperidyl]-1-(2,3,4,5-tetrahydrochysene-1H-1-benzazepine-8-yl)-1-acetone, irinotecan, (S)-N-ethyl-N-methyl-3-[(1-dimethyl-amino) ethyl] carbanilate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol methyl carbamate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-carbanilate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-3a, 8-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-8-benzyl-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, (-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate also, (-)-(3aS)-3a-methyl isophthalic acid, 2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-4 '-isopropyl phenyl carbamate, 1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol-N-2 '-methyl phenyl carbamate, (-)-(3aS)-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, (-)-(3aS)-1-phenethyl-3a, 8-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, (-)-(3aS)-8-benzyl-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, (-)-(3aS)-1,3a-dimethyl-1,2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, (-)-(3aS)-1,8-dibenzyl-3a-methyl isophthalic acid, 2,3,3a, 8, the 8a-hexahydropyrrolo is [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate also, (-)-(3aS)-3a-methyl isophthalic acid, 2,3,3a, 8,8a-hexahydropyrrolo also [2,3-b] indole-5-base-N-2 '-methyl phenyl carbamate, ±-uncle's 5-[2-fourth amino-1-hydroxyethyl]-position-penylene two-dimethylcarbamate, N, N, N-trimethyl-3-[(dimethylamino) formyloxy] aniline, ambenonium chloride, 1-methyl-3-pyridone father-in-law dimethyl carbamate, 1-naphthyl methyl carbamate, 2-isopropoxy benzene-N-methyl carbamate, 2,3-dihydro-2,2-dimethyl benzofuran-7-phenol methyl carbamate, adjacent cumenyl methyl carbamate, a toluene-N-methyl carbamate, benzimidazolyl-2 radicals-aminocarbamic acid methyl ester, 9-amino-1,2,3, the 4-tetrahydro acridine, 7-methoxyl group tacrine, 11-methyl-3-methoxyl group-4a, 5,9,10,11,12-six hydrogen-6H-benzofuran [3a, 2-ef] [2] benzo-aza-6-alcohol, (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also, 7-methyl-5,6,9,10,11,12,13,14-octahydro-5,11-tetrahydropyridine-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also, 12-amino-3-chloro-9-ethyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline, 12-amino-3-chloro-9-methyl-6,7,10,11-tetrahydrochysene-7,11-methano ring suffering [b] quinoline, N-[2-hydroxyl-3-methoxyl group-5-chloro-benzal]-huperzine A, 12-(2 '-carboxyl-5 '-anisyl)-2,12-dihydroxy-dodecane-4-ketone, O, O-DFP, N-(phosphoric acid methyl) glycine, O, O-dimethyl-(2,2,2-three chloro-1-hydroxyethyls) phosphate ester, O, O-dimethyl-O-(2, the 2-dichloro) vinyl phosphate, O, O-dimethyl-O-p-nitrophenyl thiophosphate, O, O-dimethyl-S-(N-methylamino formyl methyl) phosphorodithioate, O, O-dimethyl-S-(N-methylamino formyl methyl) thiophosphate, O, O-dimethyl-S-[2-(1, the 4-diethyl succinate) base] phosphorodithioate, Phenylmethanesulfonyl fluoride, methanesulfonyl fluoride, tetra isopropyl pyrophosphoramide, Fasciculin-I, Fasciculin-II or its pharmaceutically acceptable salt.
3. purposes as claimed in claim 1 is characterized in that, described acetylcholinesteraseinhibitors inhibitors is selected from down group:
(±)-2-[(1-benzyl-4-piperidyl) methyl]-5,6-dimethoxy-1-Indanone;
3-[1-(benzyl)-4-piperidyl]-1-(2,3,4,5-tetrahydrochysene-1H-1-benzazepine-8-yl)-1-acetone;
(S)-and N-ethyl-N-methyl-3-[(1-dimethyl-amino) ethyl] carbanilate;
1,2,3,3a, 8,8a-six hydrogen-1,3a, 8-trimethyl pyrrolo-[2,3-b] indole-5-phenol methyl carbamate;
9-amino-1,2,3, the 4-tetrahydro acridine;
11-methyl-3-methoxyl group-4a, 5,9,10,11,12-six hydrogen-6H-benzofuran [3a, 2-ef] [2] benzo-aza-6-alcohol;
(5R, 9R, 11E)-and 5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone also;
O, O-dimethyl-(2,2,2-three chloro-1-hydroxyethyls) phosphate ester;
The tetra isopropyl pyrophosphoramide;
Fasciculin-I or Fasciculin-II;
And pharmaceutically acceptable salt.
4, purposes as claimed in claim 1 is characterized in that, described inhibitor be (5R, 9R, 11E)-5-amino-11-ethidine-5,6,9,10-tetrahydrochysene-7-methyl-5,9-methylene ring suffering is (b) pyridine-2 (1H) ketone and pharmaceutically acceptable salt also.
5. as the described purposes of claim 1-4, it is characterized in that described medicine contains the acetylcholinesteraseinhibitors inhibitors and the pharmaceutically acceptable carrier of 0.005-99 percentage by weight.
6. purposes as claimed in claim 1 is characterized in that described diabetes are type i diabetes.
7. purposes as claimed in claim 1 is characterized in that, described medicine is tablet, granule or capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510027127 CN1709508A (en) | 2005-06-24 | 2005-06-24 | Use of acetylcholinesterase inhibitor for preparing diabetes medicines |
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| CN 200510027127 CN1709508A (en) | 2005-06-24 | 2005-06-24 | Use of acetylcholinesterase inhibitor for preparing diabetes medicines |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8093262B2 (en) | 2005-05-23 | 2012-01-10 | President And Fellows Of Harvard College | Use of huperzine for disorders |
| EP2203169A4 (en) * | 2007-09-18 | 2013-12-04 | Stephen Wills | Glycemic control, diabetes treatment, and other treatments with acetyl cholinesterase inhibitors |
| CN116059207A (en) * | 2023-03-15 | 2023-05-05 | 浙江大学 | Application of huperzine A in reducing blood sugar and preventing and treating diabetic nephropathy |
-
2005
- 2005-06-24 CN CN 200510027127 patent/CN1709508A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8093262B2 (en) | 2005-05-23 | 2012-01-10 | President And Fellows Of Harvard College | Use of huperzine for disorders |
| US8193212B2 (en) | 2005-05-23 | 2012-06-05 | President And Fellows Of Harvard College | Use of huperzine for neuropathic pain |
| US8822492B2 (en) | 2005-05-23 | 2014-09-02 | President And Fellows Of Harvard College | Use of huperzine for disorders |
| EP2203169A4 (en) * | 2007-09-18 | 2013-12-04 | Stephen Wills | Glycemic control, diabetes treatment, and other treatments with acetyl cholinesterase inhibitors |
| CN116059207A (en) * | 2023-03-15 | 2023-05-05 | 浙江大学 | Application of huperzine A in reducing blood sugar and preventing and treating diabetic nephropathy |
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