TW200946119A - 1,2,3,4-tetrahydroisoquinoline derivatives, manufacturing method thereof, and pharmaceutical compositions containing the same - Google Patents

Abstract

The present invention provides 1,2,3,4-tetrahydroisoquinoline derivatives, the derivatives have the structure of the following formula (I), wherein R1 is propargyl or cyclopropylmethyl, while R2 is a substituted group of N-Ethyl-N-methylamino or 1-Pyrrolidyl or 1-Piperidinyl or 1-Morpholinyl. The present invention also provides the manufacturing method thereof, and the pharmaceutical compositions containing the same. The derivatives can be applied in the process of protein hydrolysis to regulate amyloid precursor proteins (APP), thereby further providing the novel compound to treat Alzheimer's disease (AD).

Description

200946119 IX. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a novel 1,2,3,4-tetrahydroisoxoquinoline derivative, in particular to a treatment for Az A 1,2,3,4-tetrahydroisoquinoline derivative of Haimo, a preparation method thereof, and a pharmaceutical composition containing the same. ^ [Prior Art] Alzheimer's disease (AD) is a disease of paralytic neurological loss due to the deposition of non-hydrolyzable toxic proteins in the brain. When the German neuropathologist (S Alzheimer) made a brain anatomy of a woman with severe dementia in the first (four) world, Wei _ 敝 敝 — — — — — — — — — — — — — — — — — — — 异常 异常 异常 异常 异常 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 The research towel found that the more _ from the non-water eggs from the f group of red will cause inflammation of the brain nerve cells and society. A _ 症 也 is also named after the name of the pathologist of Qiqi. Alzheimer's disease is mainly caused by the damage of nerve cells in the hippocampus of the cerebral cortex. The city causes patients to gradually lose their memory and have language and emotional disorders. The early symptoms are forgetfulness, and the daily manifestations are progressive mosquitoes. Qian Huo ★ her position (four) to help the shirt turn. Late gestational weight _ knowing obstacles, (4) adapting to social life, leading to the inability to understand, the axis of the _ energy domain, the problem of the butterfly, and eventually ill in bed. A _ 默 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身 本身The death of the dead life 200946119 life. Statistically, 'women are more susceptible than men, 11% of the population over 65 years old have an opportunity to suffer, and about 50% of people over 80 years old have the opportunity to suffer from this disease, so it is also known as Alzheimer's disease. One of the pathological features of morbidity is the deposition of amyloid plaques (amyi〇id piaqUe) in the brain, and these plaques are mainly composed of β-type amyloid peptides (j3amyl〇id_peptides, Ap). The peptide (Αβ) is not a direct product of the gene, but a product obtained by cleavage of a previously produced starch-like precursor protein by a different enzyme. The amyk precursor protein (APP) is a single transmembrane. Protein, which has two pathways of decomposition: one is sequentially cleaved by α-type secretase (a_secretase) and γ-secretase (γ-secretase) to produce a soluble protein-like protein (a soluble amyi〇id) Precurs〇r pr〇tein a, sAppa); another path is After being cleaved by the β-type secretase (p_secretase) and then cleaved by the sputum-type secretase, an insoluble β-type amyloid peptide (Αβ) can be produced. In the brain cells of patients with Alzheimer's disease, the proteolytic process of the prodrug protein (ΑΡΡ) of the scorpion venom may pass through the decomposition pathway of p-type secretase and sputum secretase, resulting in more soluble ρ type powdered peptide (10)). Because the sputum-type secretase has different cutting points on the precursor powder of the genus powder, it will produce different sizes of soluble β-type phenotype peptide _, which has strong toxicity and easy fibrosis. The polymer is formed and accumulated as the core of the plaque, so that other types of p-type powder-powder peptides _ (large-part Αβ4〇) are more likely to accumulate on these plaques. At present, in the brains of patients with Alzheimer's disease, Αβ42 accounts for a higher proportion of all Αβ than normal people. By cell layer and pharmacological studies, it was found that γ-type secretase is an aspartic acid protease (IV) _, and the regulation of γ-type secretase is a therapeutic target, which is considered to be very important in the treatment of Alzheimer's disease. direction. However, another therapeutic direction is directed to the proteolytic process that regulates the secreted enzyme 200946119. The process is to cleave the starch-like precursor protein (APP) by her-secretase, and then produce a soluble starch-like precursor protein a (sAPPa), thereby blocking the insoluble p-type amyloid peptide. (Αβ) is produced. The current study found that the proteolytic pathway of alpha-secretase can be regulated by the mitogen-activated protein kinase (ΜΑΡΚ) pathway. The μαρκ ' pathway is related to the activation process of a series of mitogen-activated protein kinases (mitogen_activate(i prOtein 'kinase, ^Κ) and extracellular signal_fegulated ❹ protein kinase (ERK). Therefore, how to regulate ΜΑρκ information The MAPK signaling cascade, which in turn affects the proteolytic process of alpha-secretase, is also important in the treatment of Alzheimer's disease. The development of drugs for patients with early Alzheimer's disease mainly uses drugs to relieve early Aziz Symptoms of patients with Alzheimer's disease. The US Food and Drug Administration (FDA) has approved four drugs to improve memory and slow the progression of Alzheimer's disease. The first drug is approved Tacrine. The trade name is Cognex), which has many side effects, including hepatotoxicity and poor performance in improving memory. The other three new drugs are DonepezH (trade name is Aricept). Rivastigmine (trade name is Resin (Exel〇n) and galantamine (galantamine 'trade name is profit Reminyi, these drugs have been shown to improve memory and have fewer side effects. Taknin and D〇nepezii both block the activity of cholinesterase by blocking the activity of cholinesterase Inhibition of the breakdown of acetyich〇nne. Both drugs increase the amount of acetylcholine in the brain to delay the loss of memory and help patients perform the daily activities required for daily living. Failure to cure Alzheimer's disease can only alleviate the symptoms of Alzheimer's disease in 200946119 and the drugs are not effective for everyone. The efficacy is limited to patients with early and mid-term Alzheimer's disease. Therefore, prevention and early diagnosis Zhaimer's disease is still the current medical bottleneck. It can be seen that there are still many missing items in the above-mentioned household items, which is not a good designer, but needs to be improved. Soluble starch precursor protein α (sAPPa) is known. It has neurotropic and neuroprotective activities against oxidative (〇xidative) and excitotoxic damage. B-type monoamine oxidase B (MAO-B) inhibitor rasagmne (rasammne), and rasagiline contains carbamyl-containing derivative TV_3326 in neuroblastoma (_ T_) SH-SY5Y and PC12 can inhibit the protein-cleaving process of starch-like precursor protein (APP) via mitogen-activated protein kinase (K), thereby stimulating neuroprotective soluble starch-like precursor egg (IV) alpha (sAPPa) Interpretation ^ and another - well-known b-type monoamine oxidase shack warship Wei Jilan, in a variety of clinical dragon-style bio-finance research can now increase left

Spinosa Levodopa) to cause neuroprotective effects to treat Parkinson's disease ins〇n, s cii mine). This sputum (4) acid sister Geeland and Lai Ji Cai rely on the path (MEK-dependem pathways) to increase the soluble powder-like precursor protein ^ (10) called release. In summary, the secretion of _ecretase regulates the starch-like precursor protein (10). The river has two protein cleavage pathways, which affect the flour pro-expressing APP) proteolytic pathway by regulating intracellular pathways and extracellular signal-regulated proteins. The starch-like precursor protein (僧) protein cleavage pathway is oriented toward the non-amyloidogenic pathway regulated by the type secretase 200946119 (n amybidGgeme pathway), which is produced by the sputum, or by inhibition of 丫Type of secretase activity to reduce the production of insoluble β-type amyloid peptide (Αβ) 'Keco-inducible starch precursor protein (4)) protein cleavage process toward the "type of sputum enzyme regulation"; The path (__office hall pharyngeal pathway) to produce the sappa of mineral powder pre-white matter a (sAppa) with the help of God, ., U. Therefore, by directly or indirectly controlling the gamma-type shouting live silk _ powder precursor protein f (App) protein cleavage pathway can be used as a new direction for the treatment of Alzheimer's disease. m Yes, the inventors of the present invention in view of the above-mentioned disadvantages of the use of Alzheimer's disease drugs, is thinking Improve and innovate 'and After many years of painstaking research, I finally succeeded in research and development of this 1, 2, 3, 4-reading and reading organism, its preparation method and pharmaceutical composition containing the same. [Invention] The object of the present invention is to provide a kind of 1,2,3,4_ton of heterogeneous derivative, chemically modifying 1,2,3,4-tetrahydroiso-light" through a chemical synthesis step, finally obtaining a novel (2) fine hydrogen having the structure (1) © structure The present invention is based on the provision of a method for preparing a novel 1,2,3,4-tetrahydroisoquinoline derivative having the structure of formula (1), Wherein R1 and the illusion are defined. The other object of the present invention is to provide a pharmaceutical composition comprising a ruthenium derivative which comprises a structure of the formula (1). Hydrogen isosine derivatives for the treatment of Alzheimer's disease, in which R1 is as defined in the text. $ Know some type B mono- oxidase B (ΜΑΟ-Β) inhibitor package 9 200946119 Reza Rasagiline, Rezaghilan contains carbamylc〇ntaining derivative* 326 and seiegiiine can regulate the protein-like proteolytic protein (App) protein cleavage process via ERK-related pathway, thereby reducing the production of insoluble β-type peptide peptide (Αβ) , and extracellular signal-regulated protein kinase (ERK) activation can indirectly regulate the release of soluble starch-like precursor protein a (sApPa). Therefore, these type B monoamine oxidase inhibitors may be used to treat Alzheimer's disease. It is known that these b-type monoamine oxidase inhibitors have a partial chemical structure of propargylamine, and it has been found that this structure is important for the activation of ERK signals, but is independent of inhibition of type B monoamine oxidase activity. In addition, conventional acetylcholinesterase inhibitors can be used to treat Alzheimer's disease. Such acetylcholinesterase inhibitors often have an amine carbachyl group (carb_yl). In order to achieve the above object, the inventors of the present invention used commercially available tetrahydroisoquinoline (U,3,4-tetrahydr〇iS〇qUin〇line) as a reaction starting material (3), which was different by chemical synthesis reaction. Modification of the functional groups to produce novel 12,3,4-tetrahydroisoquinoline derivatives. By chemical steps (a) to (7), a novel derivative of 1,2,3,4-tetrahydroisoquinoline, lla_d & 12a d, which is a derivative having the structure of formula (I) as shown in Figure 1, can be obtained, respectively. , where R1 and R2 are as defined in the text. Therefore, the present invention uses the chemical synthesis method in the first embodiment to modify the u,3,4-tetrahydroisoquinoline to have a German structure with the B-ammonium secretase inhibitor, and the The acetylcholinesterase inhibitors have similar chemical structural compositions (amine-methyl sulfhydryl groups), which in turn may play a role in the activation of ERK and the release of soluble starch-like precursor protein alpha (sAppa). It is known that the activation of ERK pathway (IV) inhibits the activity of sputum-type secretase, and further investigates whether these derivatives have an effect on the activity of Y-type 200946119 secretase. [Embodiment] The present invention provides a 1,2,3,4-tetrahydroisoquinoline derivative and a pharmaceutical composition comprising the same, wherein the derivative and the pharmaceutical composition containing the same comprise a compound of the formula (1), The towel R1 has a structure of the formula (Π) as 'b, 2, 3 or 4. In a preferred embodiment, m is a R1 ^ is a propynyl group ((4) (four) anti-1); or R1 Having a structure such as a circle (ΠΙ), η is a b, 2, 3 or 4, and in the preferred embodiment η is 1, then ri is a CyCi〇pr〇pyimethyi. Wherein R 2 has the structure of the formula (IV), and R 3 is selected from the group consisting of methyl (even), ethyl (CH 2 CH 3 ) or C & (-(CH 2 ) 2 CH 3 ); R 4 # iSi f ^ (-CH 3 ) 6^(CH2CH3)^^^(-(CH2)2CH3) > In a preferred embodiment, R3 is a fluorenyl group (even) and R4 is a methyl group (even), which is like nethyl-N -Methylamino (N_Ethyl_N_methylamin〇); or a structure having the formula (7) of Figure 1, wherein X is selected from the group consisting of even-, oxygen or sulfur. In a preferred embodiment, the χ is _, then the illusion is ^ Ordinary bite (1-PynOlidyl); or R2 has the structure of formula (VI), wherein γ is selected from rhyme 2_, β oxygen or sulfur, and in the preferred embodiment, 丫 is even _, then illusion is u Peach-biting base (4) e-na or R2 has the structure of formula (5), wherein γ is selected from the group consisting of even-, oxygen or sulfur. In a preferred embodiment, Y-Wei, R2 is 1_Ferrinyl (1 Yang) Μ Μ γ χ χ χ _ 之 } , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , 适 适 适 适 适 适 该 该 该 该 该And the disease that causes the nerves to die, reduce the memory and learn the loss of the Wei, and the process of the silk and its caps. The disease in which the cognitive process is impaired includes, but is not limited to, Alzheimer's disease or other dementia 11 200946119, or Parkinson's Disease. Such excipients include, but are not limited to, diluents, fillers, binders, disintegrants, slip agents, and the like. Wherein the excipient includes, but is not limited to, microcrystalline cellulose, polyvinylpyrrolidone (PVP), corn granules, modified starches, and rebellious temples. Sodium starch glycolate), resin, gelatinized starches, • sugars, polyethylene glycol (PEG), polyviny丨alcohol, hydroxypropyl cellulose , methylcellulose, hydroxymethyl cellulose, hydroxypropyl m methylcellulose, etc. The present invention is exemplified by the following examples, but the present invention It is not limited by the following examples. EXAMPLE I Chemical Synthesis of 1,2,3,4-Tetrahydroisoquineline Derivatives As shown in Figure 1, the present invention is 1,2,3,4-tetrahydroisoindole φ (l, 2, M -tetrahydroisoquinoline) as the starting material for the reaction (3) (purchased from Sigma-Aldrich, USA)

Co.) 'modification of different functional groups by chemical synthesis reaction to produce a novel lysine derivative having the structure of formula (1).

(I) The chemical synthesis steps of the novel 1,2,3,4-tetrahydroisoquinoline derivatives of the present invention are as follows: 12 200946119 Step (a): N-trifluoroethenyl-1,2,3 Preparation of 4-tetrahydroisoquinoline (4) Dissolve 1,2,3,4-tetrahydroisoquineline (3) (8 ml, 60 mmol) in dioxane (ch2C12) solution and drip into ice cold A solution of dichloromethane (32.4 ml) was added and the solution contained trifluoroacetic anhydride (CF3CO) 20 (9 ml), followed by stirring for 3.5 hours in an ice-cold environment. Further, a solution of potassium hydroxide (KOH) (4.74 g, 85 mmol) dissolved in water (72 ml) was added to the above solution, and stirred at room temperature for two hours. After extracting the mixture with di-methane, sulfuric acid was used. Magnesium• (MgSCU) dry organic layer. Finally, the residue was purified on a silica gel chromatography column (siUcagel c〇lumn) (n-hexane/ethyl acetate (EtOAc) = 3:1 as eluent) to obtain 9.92. g N-Trifluoroacetyl-l,2,3,4-tetrahydroisoquinoline (4) in yellow oil (yield 72%) ). Thin layer chromatography (TLC) / ^ = 0.58 (n-hexane / ethyl acetate = 1 : </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; m,4H, Aryl calendar); 4.80 (s, 2H, Ph - CH2 - N); 3.91 - 3.86 (m, 2H, Ph - CH2 - CH2 - N); 3.00 (br s: 2H, Ph - CH2 - C /^2 - N) 'After Fast At〇m ❹ Bombardment Mass Spectrometry (FABMS), the molecular formula is ChHhjNOFs, m/z: 230 (M+ 1). Step (8): 7-chloroacetamidine - Preparation of trifluoroethyl fluorenyl-1,2,3,4-tetrahydroisoquinoline (5) 'Dilute chloroacetyl chloride (15 ml) into 2 g of dioxin (62 ml) In the suspension, vaporized aluminum (AICI3) (24.16 g) was dissolved in the suspension at a temperature of 〇 to 5 〇c, and after argon atmosphere for 20 minutes, the suspension was allowed to stand at room temperature. The intermediate N_trifluoroacetamido-1,2,3,4-tetrahydroisosalin (4) (9.29 g, 40 mmol) was added to the mixture and allowed to act at room temperature 13 200946119 over 30 After the mixture was stirred for an additional 3 hours, it was added to ice water (407 ml). After stirring for 5 hours, the layer was separated and the aqueous layer was extracted with dioxane (2 X 200 ml) using water (2 X 240 ml) and 5% aqueous sodium hydrogen carbonate (NaHC03) (3x 240 ml) washing organic layer. After drying the organic layer, the solvent is evaporated to dryness to obtain 1〇47g of white solid Zhao 7-gas acetyl trifluoroacetamido-1,2,3,4-tetrahydrogen Isoquinoline (7-Chloroacetyl-N-trifluoroacetyl-l, 2,3,4-tetrahydroisoquinoline) (5) (yield is * 88%). Thin layer chromatography (TLC) outside = 0.4 (positive hexane / acetic acid g==3:1).mp: 120-122 0C. 0 Alignment by 11 d11 (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 7.78 (q, 4 Hz) ,1H,Aryl Η),7·34 - 7.26 (m,2H,Aryl),4.87 (s, 2H, Ph-Ci/2 - N), 4.67 (s, 2H, - CH2 - C=0), 3.95 - 3.87 (m, 2H, - CH2 - N), 3_06 - 3.01 (m, 2H, Ph-C/f2_) 'After fast atom bombardment mass spectrometry (FABMS), the molecular formula is CnHuNC^ClFij, /w/ z: 306 (Μ + 1). Step (c): Preparation of 7-chloroethyloxy-indole-trifluoroethyl fluorenyl-1,2,3,4-tetrahydroisoquinolinyl 7-chloroethyl-indole-tris-acetyl Base-1,2,3,4-tetramine (5) (8.4 g, 27 mmol) dissolved in anhydrous dioxane (58 ml), then 3-oxoperoxybenzoic acid (3-chloroperoxybenzoic acid, mCPBA) (12.3 g) - all added to the solution. The suspension was cooled to 〇 ° C, and trifluoroacetic acid (2.0 ml) was added dropwise to the ice-cold suspension for more than 5 to 10 minutes. The reaction flask was shielded from light and stirred at room temperature for 3 to 5 days, poured into water (1 ml), and neutralized using an ammonium hydroxide solution. The layers were separated and the aqueous layer was extracted with di-methane (60 mL). Finally, it was purified by a silica gel chromatography column (using Zhengjiyuan/ethyl acetate=2:1 as eluent) to obtain 4.24g of 200946119 yellow oily 7-gas ethoxyl-N-trifluoroethane. 7-Oiloroacetoxy-N-trifluoroacetyl-l (2,3,4-tetrahydroisoquinoline) (6) (yield 48%). Thin layer chromatography (TLC) = 0.5 (n-hexane / ethyl acetate = 3:1). Alignment was determined by 1H NMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 7.20 (t, J = 7 Hz, 1H, Aryl H), 7.01 - 6.93 (m, 2H, 2H) , Aryl Hs), 4.75 (s, 2H, • Ph-C//2-N), 4.33 (s, 2H, CO - CH2 - ), 3.90 - 3.82 ( m, 2H, N-CH2-), ' 2.97 — 2.92 (m, 2H, Ph· C//2 -), and the molecular formula is C13HUN03C1F3, w/z: 322 (M+ 1) by fast atom bombardment mass spectrometry (FABMS). Step (d): Preparation of 7-hydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (7)

For the fermentation of methanol (MeOH) containing 7-gas ethoxylated-N-trifluoroethylidene-1,2,3,4-tetrahydroisoquineline (6) (3.6 g, 11 mmol) In the solution, 15% aqueous sodium thiomethoxide (NaSCH3) (10 ml, 1 〇 7 mmol) was added at 20 ° C for 2 hours, and then the mixture was acidified to pH with 3N hydrochloric acid (HC1). Hey. 7-Hydroxy-l,2,3,4-tetrahydroisoquinoline hydrochloride salt (7) ❹ (yield 54%) ϋ.2 (2 qijia entertainment) y methanol = 5:1). Alignment was determined by iHNMR (d_s〇lvent: heavy water (〇2〇)) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 7.15 - 7.00 (m, 3H, -2H, Aryl Hs), 4.22 (d, J = 6 Hz, 2H, Vh-CH2 - N), 3.80 - 3.58 (m3 2H, ' Ph_c//2 - CH2 - N), 2.65 (t, 5.4 Hz, 2H, Ph-C//2 - CH2 - N Then, by the fast atom bombardment mass spectrometry (FABMS), the molecular formula is c9HnNO.HCn, w/z: 186 (M+ 1). 7-Methyl-l,2,3,4-tetrahydroisoquinoline hydrochloride (7) using propynyl bromide, respectively, has a partial structure or a cyclopropanemethyl group as shown in FIG. Bromo 15 200946119 (cyclopropylmethyl bromides), having a partial structure of the formula (melon) as shown in Fig. 1, performing an N-alkylation reaction in an experimental environment, and performing step (8), respectively, to obtain a 7-base group _N_propynyl-1,2,3,4-tetrahydroisoquinoline (8) or 7-carbyl-N-ring 7-Hydroxy-N-cyclopropylmethyl-1,2,3,4-tetrahydroisoquinoline (9) 〇♦ Step (e-1) : 7 -Hydroxy-N,propynyl-1,2,3,4-tetrahydroisoquinoline (8) Preparation of 7-hydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (7) (1.7 g, 11 mmol) and potassium carbonate (KAO3) (3 g, 22 mmol) stirred in a solution of methyl cyanide (CH3CN) (33 ml), then 80% propynyl bromide (1.3 ml) , 11 mmol) solution was added to the above solution. The reaction mixture was stirred for 25 hours under a nitrogen atmosphere and filtered. After the filtrate was washed with water, the aqueous layer was extracted with ethyl acetate (2×40 ml). The organic layer was dried over anhydrous magnesium sulfate (MgSO.sub.4), filtered and concentrated in vacuo. Purification, 110 mg of 7-hydroxy-N-propynyl-1,2,3,4-tetrahydroisoquinolinium (7-Hydroxy-N-propargyl-1,2,3,4) -tetrahydroisoquinoline) (8) (yield 50%). = 〇·4 (dichlorocarbonitrile / methanol = 9:1). Mp: 82 - 83 0C. Alignment was determined by 1H NMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 6·91 (d, "/= 8.1 Hz, 1H, Aryl H), 6.60 (d, 8·) 1 Hz, 1H, Aryl H), 6.38 (s, 1H, Aryl H), 3.69 (d, */= 23 Hz, 2H, Ph-CH2 - N), 3.51 (d, / = 1.8 Hz, 2H, N - CH2 - C = C - ), 2.84 (t, J = 4 Hz, 4H, Ph-CH2-CH2-N), 2.31 (t, "/= 2 Hz, 1H, CeCH), followed by fast atom bombardment mass spectrometry The technique (FABMS) was found to have the formula C12H13NO, w/z: 188 (M+ 1). Step (e-2): Preparation of 7-hydroxy-N-cyclopropanemethyl+2,3,4-tetrahydroisoquinoline (9) 200946119 First 7-hydroxy-1,2,3,4-tetra Hydrogen isoquinoline hydrochloride (7) (0.9 g, 6 mmol) and potassium carbonate (2.0 g, I2 mmol) were stirred in a solution of decyl cyanide (18 mi) and then 96% (bromo) A solution of methyl bromomethyl cyclopropane (0.6 ml, 6.3 mmol) was added to the above solution. The reaction mixture was continuously mixed for 25 hours under a nitrogen atmosphere. The filtrate was washed with water and the water layer was extracted with ethyl acetate (20 ml X 2). The organic layer was dried using anhydrous magnesium sulfate (MgS 〇 4). The residue was purified on a silica gel chromatography column (digested with 2 gas / methanol = 9:1) to obtain 0.37 g. 7-Hydroxy-N-cyclopropylmethyl-l,2,3,4-tetrahydroisoquinoline (9) (Yield 57%). Thin layer chromatography (TLC) i?f = 0.3 (dichloromethane/methanol = 9:1). Mp: 11δ - 119 0C. Alignment was determined by if! NMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 6.87 (d, ·/= 8.1 Hz, !H, Aryl H), 6.58 (dd, */= 8.1 Hz, 2.1 Hz, 1H, Aryl H); 6.24 (d, J = 2.1 Hz, 1H, Aryl H), 3.58 (s, 2H, Ph-C^2 - N), 2.87 (dd, 5 Hz, 4H, Ph-C//2 - CH2 - N), 2.48 (d, / = 7 Hz, 2H, N - CH2); 1.13 - 1.10 (m, ❹1H); 0.6 Bu 0·57 (m, 2H ), 0.21 (q, (8 Hz, 2H) ' is again known by fast atom bombardment mass spectrometry (FABMS) with the formula C13H17NO, w/z: 204 (M+ 1). Finally, 7-hydroxy-N-propynyl -1,2,3,4-tetrahydroisoquineline (8) and 7-hydroxy-N-cyclopropanylmethyl ' _1,2,3,4-tetrahydroisoquinoline (9)' were respectively added to N,N-di A similar compound (R2-CO-C1) (l〇ad) in a methyl cyanide solution, where R2 is N-ethyl-N-methylamine, respectively. N-Ethyl-N-methylamino, ln-Pyrrolidyl, 1-_piperidinyl or ^Ferrinyl (丨啡啡^(四)); 17 200946119 a structure having the formula (IV), (V), (VI) or (VH) as shown in Fig. 1; The compound (R2-CO-C1) (10a-d) is N-ethyl-N-methyl-carbamoyl chloride (10a), respectively. Pyrrolidine carbonyl chloride (10b), piperidinel carbonyl chloride (10c) and morpholine carbonyl chloride (lOd), after step (i), respectively 1,2,3,4-tetrahydro-isoquinoline derivatives lla-d and 12a-d. The above N-ethyl Ν-methyl-amine formazan, (N-ethyl-N-methyl- Carbamoyl chloride) (10a), pyrrolidine carbonyl chloride (10b), piperididine carbonyl chloride (10c) and morpholine carbonyl chloride (lOd), None of them can be obtained from the market. Therefore, the synthesis of 10a-d can be carried out by using the literature (1) conventional method 'by using a toluene solution containing phosgene. Step (f-Ι): Preparation of a derivative of 1,2,3,4-tetrahydroisoquinoline lla „ d is a common step of first 7-carbyl-N-propyl block-1, 2, 3, 4-θ-hydrogen isoindene (8) (1 equivalent of hydrazine in a semi-soluble solution in a methyl alcohol solution, mixed uniformly into an ice-cold mixed solution, and then a similar compound called di-filament amine (R2-CO-C1) (l 〇ad), separately added to each of the above-mentioned ice-cold mixed solutions (four groups in total), the scramble is uniform, and then 60% sodium hydride (60% of the surface is dissolved in mineral oil, u equivalent) is added to the four groups in drops. The four groups of reaction mixtures in the mixture were respectively allowed to stand at room temperature and air atmosphere for 2 hours. After the solvent was volatilized in a vacuum, water was added and extracted three times with acetic acid B. _ layer dry work, volatile field drying in vacuum. Purification of residue gelatin chromatography potential column 'can be made four (four), 3,4_ read heterogeneous derivative ((4) final trace step (four): preparation (8) The common step of the derivative of cold tetra-argon isoquinoline 仏-d is 200946119. First, the solution of 7-based-N-cyclopropanone methyl ketone is dissolved in methyl cyanide (9) equivalent solution. / Valley liquid, Kunming evenly becomes an ice cold In combination with the solution, the N,N_dichdylamine argon gas similarity mouth (R2-CCM:1) (l〇a_d) was added to each of the above-mentioned ice-cold mixed solutions (four groups in total). • Uniform 'then 6 〇 0 /. Sodium hydride (6 〇 % in the oil, i 3 eq.) Add a four 纟 合物 巾 , , , , The cake was argon for 2 hours. After the solvent was evaporated in a vacuum towel, water was added and extracted three times with acetic acid. The organic layer was dried over magnesium sulfate (MgS 4) and evaporated in vacuo. Drying. Purify the residue with a column of Shishixi gum chromatography to obtain four final products of the derivative of the derivative (10). Step number (f-1-1): 7-( Preparation of N-methyl-N-ethylamine-methyl oxime)-Ν-propynyl-1,2,3,4-tetrahydroisoquinoline (lla) 7-Hydroxy-N-propynyl-1 , 2,3,4-tetrahydroisoquine (8) and N-ethyl-N-methyl-carbamoyl chloride (10a) are mixed to carry out the step (f_i) Purified with a silica gel chromatography column (with acetic acid/n-hexane = 2:1 as eluent) to obtain a propyl group and &amp; 7-(N-Methyl-N-ethylaminemethyl oxime)-N-propynyl-i,2,3,4-tetrahydroisophthalocyanine (7-ethyl hydrazine-N-methylamino) (N-Methyl-N-ethylcarbamoyloxy)-N-propargyl-l, 2,3,4-tetrahydroisoquinolin e) (lla) (yield 50%) by 1H NMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian

Gemini, 300 MHz) Determination of the alignment ' δ 7.08 (d, J = 8 Hz, 1H, Aryl H); 6.88 (d, "/= 7 Hz 1H, Aryl H), 6.80 (s, 1H, Aryl H), 3.76 (s, 2H, Ph-C^2 - N), 3.50 (d, J= 2 Hz, 2H, N - CH2 - CCH), 3.42 (q, 7 = 7 Hz, 2H, N - C//2 - C^3), 3.04 - 2.81 (m, 7H, N - CH3j Ph-C//2 - CH2 - N), 2.28 - 2.26 (m, 1H, C = CH)} 1.25 - 1.15 (m, 3H, N - CH2 - CH3); IR (KBr): v cm·1 = 3300, 2923, 1715. Then, by fast atom bombardment 19 200946119 mass spectrometry (FABMS), the molecular formula is Ci6HmN2〇2," 2 is (Μ+ υ. Step (f-1-2): 7-(ι-基洛销气丙烯The base-(8) phase hydrogen isoindole (4) is prepared by the preparation of 7-glycosyl-fluorenyl- 1,2,3,4-tetrahydroisophthalocyanine (8) and the base of the gas (pyrr〇 dirty e carbonyl chloride (10b) Mixing and carrying out the step (called, using a silica gel chromatography column (with ethyl acetate / hexanol = 2:1 as eluent) to purify, can have a propanol and 丨"pyrrolidyl 7_(丨_基咯•咬氧氧)-N-propynyl w-tetrahydroisophthalein • (7-(l-Pyrrolidmecarbamoyloxy).N-pr〇pargyl-l523,4-tetrahydroisoquinoline) (lib) (Yield 53%). Alignment was determined by 1H NMR (ds〇lvent: CDC13) (nuclear magnetic resonance spectrometer, Varian see Gemini, 300 MHz), δ 7·08 (d, ··= 8 Hz, 1H, Aryl Η), δ 6.91 (dd, J= 8

Hz, 2 Hz, 1H, Aryl H), 6.83 (d, J = 2 Hz, 1H, Aryl H), 3.81 (s, 2H, Ph-C/^), 3.55 - 3.44 (m, 6H, 2 N - CH2 - CCH), 2.94 - 2.90 (m, 4H, N - CH2 - CH2), 2.30 (t, J = 3 Hz, 1H, C^CH), 1.98 - 1.89 (m, 4H); IR (KBr): v cm'1 = 3375, 2951, 1715. Further, by fast atom bombardment mass spectrometry (FABms), the molecular formula is C17H20N2O2, w/z: 285 (M + 1) 〇Step (f-1-3): 7-(1-Phenyloxycarbonyl)-N- Preparation of propynyl-:^-tetrahydroisoquinoline (1)^ 7 7-Hydroxy-N-propynyl-1,2,3,4-tetrahydroisoquinoline (8) with hydrazine carbonyl ( Piperidine carbonyl chloride) (10c) Mixing and carrying out step (f-1), using a silica gel chromatography (acetic acid ethyl acetate / • hexane = 2:1 as an eluent) to purify 7-(1-indolylcarbonyloxy)-N-propynyl-1,2,3,4-tetrahydroisoquinoline (7-(1 -Piperidinecarbamoyl〇xy)-N -propargyl-1,2,3,4-tetrahydroisoquinoline) (llc) (yield 50%). Mp: 72-73 °C. Alignment was determined by iHNMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), δ 7.08 (d, J = 8 Hz, 丨11, Aryl H), 6.87 (dd, J = 8 Hz, 2 Hz, 1H, Aryl H), 6.80 (d, J=2 Hz, 1H, Aiyl H), 3.77 (s, 2H, 200946119

Ph-CH2 - N), 3.52 (t, 7= 15 Hz, 4H, N - CH2 - ), 2.94 - 2.85 (m, 4H, Ph-C//2 - CH2 - N); 2.28 (t, 7= 2 Hz, 1H, 〇Ξ〇Η), U3 (s, 6H,); IR (KBr): v cm'1 = 3255, 2926, 17!7. (4) Fast atom bombardment f-spectrum technique _MS) The molecular formula is C18H22N2〇2, 299 (Μ + 1). Step (Μ-4): 7 points? Folin oxy- gt; Ν _ propynyl from the preparation of the 啥 啥 ( 四 四 四 四 将 将 将 将 将 将 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 4. Tetrahydroisophthalocyanine (8) and morphine line (carbonyl chlorideMlOd) are mixed and carried out in step (4), using Shishi gum chromatography column (with ethyl acetate. / 正己烧 = 2: 1 as an eluent) purification, can be obtained with propynyl and μ whelinyl 7_ (1 phenoline oxy)- Ν-propenyl tetrahydroisoquinoline (7-(l.Mo^ Holinecarbamoyloxy)-N.pr〇pargyl.l52j3s44etrahydroisoquinolm^ (lid) (yield 82%) by 1H NMR (ds〇ivent : CDCl3) (nuclear magnetic resonance apparatus, Varian

Gemini, 300 MHz) Alignment, δ 7.10 (d, J = 8·4 Hz, 1H, Aryl H), 6.90 (dd, J = 8.4 Hz, 2.4 Hz, 1H, Aryl H), 6.81 (d, 2.4 Hz, 1H, Aryl H), 3.83 - 3.56 (m, 12H, 2 - OCH2CH2N - , Ph-CH2 - N, N - CH2 - C = CH), 2.96 - 2.92 (m, 4H, Ph-CT/2 - CH2 - N), 2.32 (t, J = 2.4 Hz, 1H, - C = CH); IR (KBr): v cm'1 = 3232, _ 2951, 1706. Further, by fast atom bombardment mass spectrometry (FABMS), the molecular formula was C17H20N2O3, w/z: 301 (M+1). Step (f-2-1): 7-(N-methyl-N-ethylaminemethanone)_N-cyclopropanemethyl-1,2,3,4-tetrahydroisoquinoline (12a) Preparation of 7-hydroxy-N-cyclopropanemercapto-1,2,3,4-tetrahydroisoquinoline (9) with &gt; 1-ethyl-:indolyl-amine formazan (N-ethyl) -N-methyl-carbamoyl chloride) (10a) After mixing uniformly, proceed to step (f-2) and purify it with a silica gel chromatography column (two gas aeration/methanol = 10:1 as eluent). 21 200946119 7-(N-Methyl-N-ethylaminemethomethoxy)-N-cyclopropanemethyl-1,2 with cyclopropanemethyl and N-ethyl-N-methylamino , 3,4-tetrazine 0S: 7-(N-Methyl-N-ethylcarbamoyloxy-N-cyclopropylmethyl-1,2,3,4-tetrahydrois oquinoline) (Ha) (yield 50%). Alignment was determined by 1H NMR (d-solvent: CDCI3) (nuclear magnetic resonance apparatus, Varian Gemini '300 MHz), δ 7.06 (d, J = 8 Hz, 1H, Aryl H); 6.84 1 (s, 1H, Aryl H 6.81 (s, 1H, Aryl H); 3.71 (s, 2H, Ph-C//2 - N); δ 3.44, 3.39 (dd, J = 7 Hz, 2H, CH3 - N - CH2CH3); 3.03 - 2.75 (m, 7H, CH3 - N - CH2CH3, Ph-CH2 - CH2 - N); 2.42 (d, J = 6 Hz, 2H, N - CH2); 1.24-1.17 (m3 3H, -CH3); ^ 1.14-0.94 (q, 7 = 7 Hz, 1H); 0.58-0.52 (m, 2H); 0.19- 0.14 (m, 2H); IR (KBr): v cm4 = 2924, 1715. The molecular formula was found to be C17H24N202, m/z: 289 (M + 1) by fast atom bombardment mass spectrometry (FABMS). Step (f-2-2): Preparation of 7-(l-pyrrolidinylcarbonyl)-N-cyclopropanemethyl-I,2,3,4-tetrahydroisoquinoline (12b) -N-cyclopropylindolyl-1,2,3,4-tetrahydroisophthalocyanine (9) is uniformly mixed with her pyrrolidine carbonyl chloride (10b), and then the above step (f_2) is carried out. After purification by a silica gel chromatography column (with a second rate of methyl or decyl alcohol = 20:1 as an eluent), 7-(1-) having a propyl propyl group and a 1-pyrrolidyl group can be obtained. Ethrolidine carbonyl)_N-cyclopropanemethyl-1,2,3,4-tetrahydroisoline (7-(1 -Pyrrolidinecarbamoyloxy)-N-cyclopropylmethyl-1,2,3,4-tetra-- Hydroisoquinoline) (12b) (yield 50%). Alignment was determined by 1η NMR (d-solvent: CDC13) (nuclear magnetic resonance apparatus, Varian Gemini, 300 MHz), § 7.08 (d, ··= 8· 1 Hz, 1H, Aryl H), 6.90 (d, J= 8.1 Hz, 1H, Aryl H); δ 6.84 (s, 1H, Aryl H), 3.80 (s, 2H, Ph-C//2 -N), 3.54 (t, J = 5.1 Hz, 2H, N - CH2 - ), 3.46 (t, J = 5.1 Hz, 2H, N - CH2 -), 22 200946119 2.92 (br s, 4H, Ph-CH2 - CH2 - N); δ 2.51 (d} J = 4.8 Hz, 2H, N - CH2 -C - ); 1.92 (br s, 4H, - CH2 - CH2 - ); δ l.〇3 (br s, 1H); 0.58 (dd9 /= 6.3 Hz, 1.5 Hz, 2H); 0.20 ( d, 1.5 Hz, 2H); IR (KBr): v cm_I = 2876, 1715. Then, by fast atomic mass spectrometry (FABMS), the molecular formula is Ci8H24N2〇2, and 3〇1 (M+^. Step number (f-2-3): 7-(1-foundation)-N_ Preparation of cyproterone methyl group from tetrahydroisoindole • 7-hydroxy-N-cyclopropanemethyl-^4-tetrahydroisoquineline (9) with piperidine carbonyl chloride (10c) Mixing and carrying out the step (f_2), purifying with a phthalocyanine chromatography column (with a second rate of thiol/sterol = 12:1 as an eluent), and having a cyclopropane fluorenyl group and a hydrazine 7-(p-pyridinium oxy)-n-cyclopropanemethyl hydrazinium (7-(l-Piperidinecarbamoyloxy)-N-cyclopropylmethyl-l, 2,3} 4-tetrahydroisoquin Oline) (12c) (5G% yield) by 1H NMR (d-solvent: CDC13) (nuclear magnetic resonance, Varian)

Gemini, 300 MHz) Alignment, δ 7.06 (d, /= 8·4 Hz, 1H, Aiyl H); 6.86 (dd, 8.4 Hz, 2.4 Hz, 1H, Aryl H)3 6.80 (d, J= 2.4 Hz, 1H, Aryl H), 3.71 (s, 2H, ?h-CH2 - N), 3.57 (br s, 2H, N - CH2 - ), 3.50 (br s, 2H, N - CH2 - ), 2.92 - φ 2.81 (m, 4H, Ph-C//2 - CH2 - N), 2.44 (d, J= 6.9 Hz, 2H, N - CH2), 1.62 (br s, 6H); 1.00 - 0.95 (m, 1H ); 0.59 - 0.53 (m, 2H); 0.20 - 0.15 (m, 2H); IR (KBr): v • = 2924, 1717. The molecular formula is also known as C19H26N2O2, w/z: Preparation of ke-I,2,3,4-tetrahydroisoquinoline (Ud) 7-Hydroxy-N-cyclopropanemethyl-i,2,3,4-tetrahydroisoquinoline (9) with After the carbonyl gas (l〇d) is mixed 23 200946119, the above step (f-2) is carried out, and the mixture is purified by a silica gel chromatography column (digas oxime / decyl alcohol = 1 〇: 1 as an eluent). 7_(1_?Ferlineoxy)-N-cyclopropanemethyl-M,1,3,4-tetrahydroisoindole (7) with cyclopropanylmethyl and 1_freflinyl can be obtained. -( 1 -Morpholinecarbamoyloxy)-N-cyclopropylmethyl-1,2,3,4-tetrahydroisoqui nolme) (12d) (yield 51%). By 1H NMR (d-solvent: CDC13) (nuclear magnetic resonance instrument,

Varian Gemini, 300 MHz) Alignment, δ 7.08 (d, 8.2 Hz, 1H, Aryl H), 6_S6 (d, J = 8.2 Hz, 1H, Aryl H), 6.81 (s, 1H, Aryl H), 3.75 - 3.57 (m, 10H, 〇- (CH2 - « CH2)2 - N - , Ph-C/T2 - N), 2.90 (t, J= 5.7 Hz, 4H, - C/i2 - N ), 2.80 ( t, J- 5.7 ^ Hz, 4H, Ph-CH2 - ), 2.42 (d, J = 6.6 Hz, N - CH2 - ), 0.98 - 0.92 (m, 1H), 0.59 - 0·53 (m, 2H) , 0_19 - 0.15 (m, 2H); IR (KBr): v cm·1 = 2924, 1717. The molecular formula is C18H24N203, m/z: 317 ίΜ + 1) by fast atom bombardment mass spectrometry (FABMS). Finally, via steps (8) to (f), a derivative ua-d having a structure of formula (j) wherein R1 and ^ are as defined herein, and 1,2,3,4-tetrahydroisoquinoline, and 12a-d. Example 2 Analysis of γ-secretase inhibition ability of 1,2,3,4-tetrahydroisoindoles and forest derivatives Due to the chemical structures of these derivatives and selegiline and rasagiline -Similar' Therefore, in this example, it was tested whether the derivatives have an activity of inhibiting γ-secretase, and thereby regulating the proteolytic process of amyloid precursor protein (APP). In the present example, the activity of γ-type secretase was measured in a T-20 cell strain using the cell-based quantitative cell-based assay disclosed in the literature (2) and the literature (3). 24 200946119 In order to measure the activity of γ-type secretase, it is first necessary to produce a stable transfected cell T-20. In the present embodiment, the following steps are described as follows: 1 · The construction of the C99-Gal4-VP 16/TO (C99-GV/TO) plastid is disclosed in the literature (2), which is briefly described below, due to the class The starch precursor protein (App) can be cleaved by α-type secretase or β-type secretase to form C83 (the C-terminus of the dinosaur powder precursor contains 83 amino acids) or C99 (the c-terminus of the precursor powder of the phyto-powder powder contains 99) A fragment of an amino acid), which is cleaved by a gamma-type secretase, produces a fragment of an intracellular domain (AICD). Using the genetic recombination technique to use App695_Gal4_vpi6 (APP-GV) as a DNA template', the template is a plastid obtained from Dr. Mark Bothwell (University of Washington, Seattle, WA), where the App6% DNA fragment is compiled (enc〇de). 695 amino acid precursor powder protein (app) protein fragment, Gal4 is the transcription factor of yeast ' and VP16 is viral transcription activator, the specificity described in the literature (2) A primer was used to perform a polymerase chain reaction (PCR) to obtain a DNA sequence fragment of C99-GaW-VP16 (C99-GV). This DNA fragment (C99-GV) was then cloned into the pCDNA5/TO vector (purchased from Invitrogen) to produce a plastid-like structure induced by tetracycline* C99-Gal4-VPl6/TO (C99- GV/TO). 2. Construction of Gal4-Luc plastids The construction method has been revealed in the literature (2). Briefly, in order to construct a vector having luciferase suitable for stable transfected cells, the pFR_Luc vector will be described. Using the gene recombination technology, the open reading frame 25 200946119 DNA sequence and its upstream Gal4 binding sequence were selected into the pBudCE4. .1 vector (purchased from invitrogen). Finally, the plastid is cleaved with a restriction enzyme, and the DNA sequence of the macrophage virus (CMV) driver on the pBudCE4_1 plastid is removed, and then self-ligation is performed to obtain Gal4 as a driver. A Gal4 promoter-driven luciferase reporter construct (Gal4_Luc) containing a zeocin-resistant screening marker t. * 3. Cell culture T-REx293 cell line was purchased from Invitrogen using DMEM medium containing 10% fetal bovine serum (FBS) and 5 pg/ml blasticidin (Dulbecco's Modified Eagle's medium) The cells were cultured in a humidified cell culture incubator at ambient temperature of 37 ° C and 5% carbon dioxide (C〇2). The T-REx293 cell line is a cell strain modified by Human embryonic kidney cells (HEK293). 4. Stably transfercted cell line producing T-20. The method for producing stably transfected cells has been disclosed in the literature (2). Briefly, T-REx293 cells are cultured in the above medium. In a petri dish of 1 cm, until the cell density exceeds 5〇%, the original medium is replaced with 8 ml of DMEM medium containing 10% fetal bovine serum, and the equivalent amount of C&quot;-GV/TO (5 pg) and GaW-Luc (5 pg) plastids were transfected into the medium with FuGENE 6 transfection reagent (purchased from Roche Applied Science). The transfected Ding-Ying-edge cells were cultured in 10% fetal bovine serum, 200 pg/ml hygromycin, 250 kg/ml zeocin and 5 pg/ml In blasticidin DMEM medium, 26 200946119 The above mixed medium is abbreviated as DMEM-ΗΖΒ. Individual cells derived from a single cell can then be isolated from a population of antibiotic resistant cells and cultured into individual cell lines. The individual cell lines described above were cultured in a 96-well culture dish containing 5 pg/ml tetracycline and 10 μL of compound E (compound E) in DMEM-ΗΖΒ medium (available from Harvard Medical). Dr. Michael Wolfe, from the Brigham and Women's Hospital, compound E is a gamma-type secretase inhibitor and is cultured for 24 hours. The cells were then lysed and added to the steady-Glo luciferase assay (spoken from Pr〇mega). The luciferase signal was read by a VictorLight _ microplate luminometer (purchased from PerkinElmer). The best stable and transfected cell line § squashing standard' is a cell line that inhibits the luciferase signal induced by tetraCyCline, so it is selected from the above cell lines. The T-20 cell line was used for the cell-based γ-secretase assay. 5. Cell-based γ-secretase assay A cell-based γ-type secretase assay using T-20 cell strain has been disclosed in the literature (2). Gamma-type secretase activity. This assay and its cell line were used in this example to measure the activity of γ1,2,3,4_tetrahydroisoquinoline derivatives for γ-type secretase activity. * Τ·20 cell strain contains 1% fetal calf serum (FBS), 200 pg/ml hygromycin, 5 pg/ml blasticidin and 250 pg/ml gilsin (zeocin) in DMEM medium (DMEM-ΗΖΒ) for culture. The T-20 cell line stably transfected with C99-GV and Gal4-Luc plastids was isolated from culture m by trypsinization and washed with Phosphate Buffered Saline (PBS). The cells were resuspended using 27 200946119 DMEM-ΗΖΒ medium, and then the suspension cells were plated at a density of 2χι〇4 cells/5〇μ!/hole in a 96-well culture dish and placed in an incubator at a temperature of 37 °C. Cultivate for 24 hours. After dissolving each of the derivatives (lla-d and 12a-d) of the present invention in 1% dimethyl sulfoxide (DMSO), DMEM-containing i pg/mi tetracycline (tetracycUne) was used. The HZB medium was diluted to a final concentration of 1 μμΜ&gt; followed by an activity assay for inhibiting γ-type secretase, and the above-mentioned respective derivatives of ίο μΜ (iia_d and 12a-d) were separately added to the respective holes of the culture • T-20 cells. In the medium, after incubation for 24 hours at 37 °C, 'the same volume of Steady-Glo luciferase test reagent (purchased from promega) was directly added to each well to terminate the reaction, using the VictorLight cold light signal microdisk reader ( Victor Light microplate luminometer) (purchased from PerkinElmer) to read the iuciferase signal values for each well. Each group of derivatives tested for inhibition of gamma-type secretase activity was a three-repeat independent test. If the stably transfected cell line τ 2 〇 was reacted with 1% DMSO which did not contain tetracycline, the activity of the read luciferase signal was determined to be 1 time. In the same reaction test (paralleltesting), a luciferase reporter gene, such as a luciferase reporter gene under the control of the CMV driver, was used as a control group in this test. In the present example, the T-20 cell line was treated with a medium containing i〇/〇DMS0 and 1 gg/ml tetracycline as a positive control group; a τ_2〇 cell line was contained with 1% DMSO. The medium was treated as a negative control group; the other groups were treated with a medium containing different compounds (10 μΜ), 1% DMSO, and 1 pg/ml tetracycline, respectively, as a test group of different compounds. . Briefly, the Τ-20 cell line was induced by tetracycline to express the C99_GV protein fragment on the cell membrane. After the C99-GV fragment was cleaved by γ-type secretase, the ACID_GV fragment was released from the cell 28 200946119 membrane, The Ga丨4 driver binding region binds to the enzyme-producing enzyme. Conversely, if the &quot;secretase activity is inhibited, the luciferase expression is reduced or not expressed. 6. Statistical Analysis Experimental data were presented as mean quantification (SD), and each derivative was tested in triplicate independent trials in each experiment. The results of the individual experiments were analyzed by single-variant analysis (〇ne crocodile fine (four) 〇f (four)). Adding a heart-shaped coffee is used to compare the results between individual compounds. In all of the examples, p &lt; 〇〇 5 is shown to be significant. 7. Results 9 In the two groups of 12a, e, and d, which were not shown in Figure 2, were reduced to the positive control group, and the 12 μ, 12, c, and d groups were able to inhibit γ-type secretase activity. About 35_72%, while the test group of rasagilme and other 1,2,3,4-tetrahydroisoquinoline derivatives reduced the activity of γ-type secretase relative to the positive control group by about 6_H. The i,2,3,4-tetrahydroisosalvin derivative having the structure of the formula (1) wherein R1 and R2 are as defined herein inhibits the activity of the γ-type secretase. ® Example 3 1,2,3,4-tetraar-isoquinoline derivative cytotoxicity test In the present invention, animal cells are tested for whether a 12,3,4-tetrahydroisoquinoline derivative has cytotoxicity against cells. 'Including, but not limited to, human embryonic kidney cells (HEK293), and other animal cells suitable for testing cytotoxicity. In a preferred embodiment, the animal cell is human embry〇nic kidney cells (HEK293) for testing whether the 1,2,3,4-tetrahydroisoquinoline derivative has cells for cells. toxicity. 29 200946119 i. Cell culture Human embryonic kidney cells (HEK293) (purchased from Invitr〇gen) 'Use penicillin containing 10% fetal bovine serum (FBS) and 0.1 mg/ml (penieinin) and strept〇mycin 2 DMEM medium (Dulbecco's modified Eagle's medium) to culture human embryonic kidney cells, T-20 as its progeny breeding cell line and γ-30 are disclosed in the literature (3), The ambient temperature is 37. (: and 5% carbon dioxide (C〇2) concentration in a humid cell incubator. 2. Cytotoxicity test T-20 cell line was plated at a density of 5 x 104 cells / 100 μΐ / hole in a 96-well plate. Each well was cultured, and different derivatives (lla_d, 12a-d) containing 10 μM were added to the culture medium of each well, and cultured at 37 ° C for 24 hours. CellTiter 96® Aqueous non-radioactive cell proliferation assay (CellTiter) 96®Aqueous Non-Radioactive Cell Proliferation

Assay) (purchased from Promega) measures the number of viable cells by the colorimetric method. This system consists of a novel tetrazolium compound 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethyl)-2-(4-sulfophenyl)- 2H-tetrazole, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium, inner salt, MTS) Electron coupling reagent is composed of phenazine methosulfate (PMS). MTS is reduced by cell biology into a formazan compound that is soluble in tissue culture medium. The absorbance at 490 nm of the ruthenium compound can be directly measured on a 96-well plate using a Synergy HT ELISA plate reader (purchased from BioTek) without additional treatment. The dehydrogenase in the metabolically active cells converts MTS into a liquid soluble formazan compound 200946119. The amount of formazan product represented by the absorbance measured at 490 nm is directly proportional to the number of active cells in the culture. In the present example, the number of viable cells was measured by the above measurement method. In this method, a solution of 2 〇 μl of MTS/PMS combined was added to each well of a 96-well culture plate 24 hours after the culture, at a temperature of 37. (: Incubation for 3 hours, followed by measurement of the absorbance of MTS converted to formazan in the survival-cell at a wavelength of 490 nm using a Synergy HT ELISA plate reader. The number of surviving cells was absorbed at a wavelength of 490 nm. The ratio is calculated as the survival rate of viable cells in 1% DMSO medium, which is defined as 100% survival. The background absorbance is defined as the absorbance measured by one cell in each well. In the present example, the T-20 cell strain was treated with a medium containing 1% DMSO and 1 pg/ml tetracycline as a positive control group; the T-20 cell strain was used as a medium containing i% DMSO. The treatment was carried out as a negative control group; the other groups were treated with a medium containing different compounds (1 μμΜ), 1% DMSO, and 1 pg/ml tetracycline, respectively, as test groups of different compounds. Statistical analysis Ο Same as described in Example 2. 4. Results. As shown in Figure 3, the progeny cell line of human embryonic kidney cells (HEK293) was Τ_2〇, and the cells survived under the treatment of each group of derivatives. Cell viability and positive control There was no significant difference between the negative control group and the rasagiline test group, and it was confirmed that there was 1,2,3,4-tetrahydroisolation of the structure of formula (I) (where RI and R2 are as defined herein). Quinoline derivatives have no significant cytotoxicity against human embryonic kidney cell line (HEK293). 31 200946119 Example 4 Analysis of inhibition ability of b-type monoamine oxidase by 1,2,3,4-tetrahydroisoquinoline derivatives in literature (4) Some of the B-type monoamine oxidase B (MAO-B) inhibitors include: rasagiline, rasamyl-comaining derivative TV-3326 and hydrochloric acid. Seiegiline can regulate the release of the amyloid precursor protein (APP) protein by the ERK-related pathway, and indirectly regulate the release of soluble starch-like precursor protein a (sApPa). Indirectly reduces the production of insoluble beta-type peptides (Αβ). Therefore, these b-type monoamine oxidase spring inhibitors may be used to treat Alzheimer's disease. These indole monoamine oxidase inhibitors have propargylamine. Partial knot The composition is, and it is found in the literature (5) that the composition is important for the activation of the ERK signal, but it is not related to the inhibition of the activity of the type B monoamine oxidase. Therefore, the present invention uses the chemical synthesis step described in the first embodiment, i. 3,4-tetrahydroisoquinoline (l'2,3,4-tetmhydmiSOqUin〇line) modified to have a chemical structure similar to selegiline or rasagiline hydrochloride and modified to have A similar chemical structure of the amine carbaryl group of the acetylch〇linesterase inhibitor, and further explored the derivatives of the starch-like precursor protein protein (lla_d and 12a_d) with similar structures to the type B monoamine oxidase inhibitor. The possible role of the process. First, B-type monoamine oxidase (MAO-B) was purified from the mouse cerebral cortex, and the activity inhibition test of these derivatives for B-type monoamine oxidase was carried out. L-Fluorescence Test Measures B-Type Monoamine Oxidase Protein Content in Rat Brain The F344/N strain of male rats was decapitated and the brain cortex was removed to obtain type B monoamine oxidase (MA〇_B). The extracted cerebral cortex was immersed in a dish of acid buffer 32 200946119 in potassium phosphate buffer (PBS), frozen in liquid nitrogen, and stored at -80 ° C for two days. Then, the frontal lobe (fr〇ntai c〇rtex) homogenate was immersed in 〇丨M* potassium phosphate buffer (PBS) (ρΗ7·4) and centrifuged at 14,000 rpm for 10 minutes to obtain the supernatant. Source of type B monoamine oxidase (MA0_B). The standard solution (standard s〇luti〇n) contains o.l potassium phosphate buffer (PBS) and albumin (albumin), which are formulated into different concentrations. 10 μΐ of each of the standard solutions of different concentration gradients was added, and 200 μL of BCA test agent was added to mix. In the same way, dilute the supernatant from the cerebral cortex and use it as a storage solution. • (stock solution), take 1 μμ from the storage solution, add 2 〇〇BCA reagent and mix it into a sample solution. The standard solution and the sample solution were then incubated together at a temperature of 37 ° C for 3 minutes. 200 μM of each solution was taken from each of the above solutions, and added to a 96-well culture dish, and the protein content was measured at 595 nm by a fluorescence microplate reader. 2. B-type monoamine oxidase (ΜΑο-β) activity inhibition test 1 U / ml horseradish peroxidase (HRP), 1 benzylamine and 50 soluble in 〇Λ Μ · After adding the % hole culture plate to the above supernatant containing B-type monoamine oxidase (ΜΑαΒ) (protein content, 8〇(4), the derivative of each test group was dissolved in DMSO to 2〇〇pg/ml, Formulated as a storage solution (salvation-soluteion), added to each hole. These mixtures were allowed to act at room temperature for 6 minutes, and analyzed by glory. The absorbance was measured and the heart was calculated by the following formula. ^ : Half The inhibitory concentration is defined in this example as the concentration required for an inhibitor to inhibit the activity of the enzyme by 5%. Inhibition rate (%) = 1 - [(experimental group 595d595 / protein - experimental group OD595) / blank Group 595D595] X1 〇〇% 33 200946119 od595: Absorbance protein at a wavelength of 595 nm: refers to the protein content of the control group. 3. Statistical analysis is as described in Example 2. Table 1 inhibition of type B monoamine oxidase (MAO-B) Active and 1,2,3,4-tetrahydroisoquinoline derivatives

Lla-d, 12a-d k) g corpse value. Compound R2 IC5〇(μΜ) logPa Rezaglan 2.1 ±0.6 2.295 11a N-ethyl-N-nonylamino 21.5 ±1.0 2.252 lib 1-0 than hazel 13.9 ± 0.5 2.235 11c 1-stone ratio bite Base 15.7 ± 2.2 2.632 lid 1-Ferrinyl 18·7± 1.6 1.567 12a N-ethyl decylamine 12.8 ± 2.0 2.688 12b I-0 piroxime 14.9 ± 1.1 2.671 12c 1-stone tb pie base 11.6 ±0.7 3.088 12d 1-Ferrinyl 18.5 ±0.8 2.033 Calculate lipophilicity by CAChev.6.1 software 4. Results are shown in Table 1, although 1,2,3,4-tetrahydroisoquinoline The derivative (lla-d) has a lower inhibitory ability than rasagllme, but the derivatives (lla-d) can inhibit the indole monoamine oxidase (ΜΑΟ-Β) when IC5〇 is 1〇-20 # ) 5% activity. However, if the propanol composition of the (3)+redented hydrogen isoindole 34 200946119 derivative (1 la-d) is replaced by a cyclopropanemethyl group (cyd〇pr〇pylme), 1,2,3 is obtained. 4-tetrahydroisoquinoline derivative (I2a_d). 1,2,3,4-tetrahydroisoquinoline derivative (12a-d) The inhibition of B-type monoamine oxidase (MA0_B) activity showed that the ability of each 12a-d derivative to inhibit B monoamine oxidase (MAO-B) activity was similar to that of lia-d. . At the same time, the lipophilicity was calculated by cAChe v 6 1 software, and it was found that the derivatives I2a-d and 11a_d have similar sizes and fat-soluble (lipop touch city) (increased log value of about 0.45), which is convenient for drug use. Preparation (generally. The fat-soluble log of the drug is about 2-5). Thus, it was confirmed that a 1,2,3,4-tetrahydroisoindolide derivative having a structure of the formula (I) wherein rj and R2 are as defined herein can be used to inhibit the activity of type b monoamine oxidase (MA0-B). Example 5 Analysis of ERK Activation Ability of 1,2,3,4·Tetrahydroisoquineline Derivatives Rasagiline or selegiline can be used to regulate ERK-related pathways (ERK-related) The activation of the pathway, and the activation of the pathway can directly or indirectly regulate the α-secretase, thereby regulating the proteolytic process of the protein precursor protein (App). Therefore, in this example, a derivative of a derivative chemical structure consisting of 1,2,3,4-tetrahydroisoquinoline and having a cyclopropylmethyl group' and an amine hydrazine group was tested. Whether the substance can significantly regulate the ERK-related pathway. 1. Cell culture is as described in Example 2. 2. 1,2,3,4-tetrahydroisoquine derivatives for ERK activation ability using anti-acidification extracellular signal-regulated kinase 1/2 antibody (anti-phospho-ERKl/S) (purchased from Cell Signaling Technology), and anti-extracellular signal-regulated kinase 1/2 antibody 35 200946119 (anti-ERK1/2) (available from Cell Signaling Technology) to measure ERK in the activated state (phosphorylation). The T-20 cell line was cultured at a density of 5 x 105 cells/well in DMEM medium containing 10% fetal calf serum at a temperature of 37. (: culture for 18 hours. These cells were then cultured in DMEM medium containing 1 pg/ml tetracycline and cultured for 18 hours at 37 ° C. Before the activation of the ERK test, the above DMEM medium containing 1 pg/ml tetracycline was replaced with DMEM medium containing 0.5% fetal bovine serum, and then the test derivative of ΙΟμΜ was added to carry out the reaction. After the reaction, the cells were placed on ice to stop the reaction. , and the medium was drained. After removing the cells, 50 mM Tris-HCl (pH 8·0), 150 mM sodium carbonate (NaCl), 5 mM ethylenediamine was used. Tetraacetic acid (EDTA), 1 mM sodium orthovanadate, 1% Triton X-100, and protease-and phosphatase-inhibitor cocktails to lyse cells. Use BCA reagents From Pierce) to measure protein concentration. Each group of cell lysates was formulated into a protein containing 50, and electrophoresed using a 12% SDS-polyacrylamide electrophoresis gel to separate different molecular weights. Protein, electrophoresis After immunoblotting, the anti-phospho- extracellular signal-regulated kinase 1/2 antibody (anti-phospho-ERKl/2) was used to identify phosphorylated (activated) ERK and anti-extracellular signal. An antibody to the kinase 1/2 (anti-ERK1/2) to discriminate the non-phosphorylated (non-activated) ERK. In this example, the T-20 cell line was contained in 1% DMSO and 1 pg/ml tetracyclic. The medium of tetracycline was treated as a positive control group; the other groups were treated with T-20 cell line in a medium containing different compounds (10 μΜ), 1% DMSO and 1 pg/ml tetracycline, respectively. Test group of compounds. 36 200946119 3. Statistical analysis of the relative activation of ERK with the 1,2,3,4-tetrahydroisoquine derivatives of Table 2 as described in Example 2 (relative to the cell group treated with the derivative) In the untreated cell group) 4 匕-activated extracellular signal-regulated kinase 1/2 (p-ERK/ERK, %) positive control group 100 Rezaglan 131 ± 14 12a 131 soil 10 12c 155 ± 28* 12d 138 士39 Note: p-ERK/ERK indicates acid-filled extracellular signal-regulated kinase 1/2 relative to non-squamized extracellular signals Section kinase 1/2. The parent values are presented as mean ± standard deviation (sd). * represents P&lt;0.05, which is significantly different from the control group. As shown in Table 2, R1 is a propylene group, and R2 is a 7-(1-indolylcarbonyloxy)-N-cyclopropanemethyl-u,3, ternary tetrahydrogen Quinoline (7-(l-Piperidinecarbamoyloxy)-N-cyclopr〇py]met^ 〇(4) pottery, 1〇μΜ's 12c test group, relative to 1〇_的雷萨吉兰(咖(四)(四) test group, the ability to activate the ancestors More favorable than the Rezagiran test group 12 times, p &lt; 〇〇 5 in statistical analysis has significant differences. The 12a and 12d test group and the Rezagiran test group have similar ability to activate ERK, confirmed to have (1) Structure (its t Ri and the ruthenium as in the text 37 200946119 Hydrogen isoquinoline derivatives have the ability to activate erk. Example 6 1,2,3,4-tetrahydroisoquinoline derivatives affect sAPP〇^ Analysis of the extent of the B-type monoamine oxidase inhibitor can directly or indirectly activate the ERK-related pathway (ERK-related pathway) to regulate the starch-like precursor protein (App) protein cleavage process, resulting in indirect 雠 soluble starch-like precursor protein a (sAPPa Released, indirectly inhibiting the production of insoluble β-type amyloid peptide (Αβ). It was confirmed in Example 5^ The tetrahydroisoquinoline derivatives 12a, c and d have the ability to activate ERK. Therefore, in the present example, the γ 3〇 cell line is 丨, 2,3,4·tetrahydroisoquinoline derivative (12a) , c and d) tests affecting the degree of release of sAPPa. 1. γ-30 cell line and its culture are shown in the literature (3) and (6), which are briefly described as follows. This paper uses ps7〇 Chinese hamster ovary cell line. (Chinese hamster ovary cell, CHO cell) 'transfected to obtain γ_3〇 stable transfected cell line. γ-30 stably transfected cell line expressed full-length amyloid precursor protein (Αρρ) and human?卩11-1〇12, and ?611-2 protein _11^11?8, 八?11-1〇12, and ?Qi-2) can be used as a precursor protein for detecting alpha-secretase-degrading amyloid (理想ρρ) ideal model cell. _ 2_ 1,2,3,4-tetrahydroisoquinoline derivative affects the degree of sAPPa release analysis will be the same as the corresponding group of cell lysates in Example 5 Protein content, electrophoresis by SDS-PAGE to separate proteins of different molecular weights, and using monoclonal antibody (6E10) of anti-Αρι ΐ7 (purchased from Chemicom Co., USA) Western blotting method (Westem brain). In this example, γ-30 cell strain was treated as a negative control group in a medium containing 1% DMSO; the other groups contained different compounds (10 μΜ), 1%, respectively. The γ-30 cell strain was treated with DMSO and a medium of 1 pg/ml tetracycline as a test group of different compounds. 38 200946119 3 · Statistical analysis Same as described in the second embodiment. 4. Results γ·3〇 stably transfected cell lines exhibit full-length amyloid precursor protein (App) and human 1 Aphla2 and 卩6112 proteins. When the 〇1 type secretase decomposes the powdered protein precursor protein (he), it produces a soluble starch-like precursor protein a (sAPPa), so it can be used to test whether the compound affects the soluble starch-like precursor protein cx (sAPPa). Released. Using the single anti-body (6E10) of Aj3l_17 to carry out the Western blotting method to detect the soluble precursor powder protein a (10), the results are shown in Figure 4A, and U, 3, 4_ tetrahydrogen is found. (10), e and d three groups of experimental groups) the amount of soluble soluble powder protein α (sApPa) protein was more than that of the negative control group, and the l2c 4 test group was compared with the Rezaglan test group. The soluble powder precursor protein a (sAPPa) detected by the group is preferred. The results of the Western blotting method were quantitatively analyzed. The results are shown in Figure 4B. (3) The fine hydrogen isosalin derivatives (10), c and d test groups can promote the release of the soluble precursor powder protein a. The 12c test group was better than the Rezagillan test group. Therefore, it was confirmed that the structure having the structure of the formula (1) (wherein R1 and the illusion are defined) can be used to promote the release of the soluble protein precursor protein a (sAppa). The present invention provides a novel lysine-1 and 3,4-tetrahydroisoindole derivative, in particular a U,3,4-tetrahydroisoindole derivative which can be used for the treatment of Alzheimer's disease, These derivatives can be used to inhibit the activity of γ-type secretase, thereby reducing the formation of Ρ-type 搬-type peptides, and the activation of the pathway to promote the production of sAPPa. It therefore reflects potential neuroprotective activity and can be used to treat the pathological mechanisms of Alzheimer's disease. 39 200946119 A novel 1,2,3,4-tetrahydroisoquinoline derivative provided by the present invention has the following advantages when compared with the aforementioned bow test and other conventional techniques: The present invention provides A novel 1,2,3,4-tetraisoindolin derivative having a conformational structure such as '(I), wherein R1 and R2, as defined herein, have been shown to inhibit γ-type secretase activity, Further, inhibition of the production of Αβ can be used to treat Alzheimer's disease. The present invention provides a novel 1,2,3,4-tetrahydroisoquinoline derivative having a structure such as the formula: (I) 'wherein R1 and R2 are as defined herein, and have been experimentally confirmed to have a relative β A mono-amine oxidase inhibitor with better soil activation and better ability to promote sAppa release can be used to treat Alzheimer's disease. The present invention provides a novel 12,3,4-tetrahydroisoquinoline derivative having a structure such as formula (I) wherein R1 and R2, as defined herein, have been experimentally confirmed to simultaneously inhibit γ-type secretase activity. In addition, inhibition of the production of Αβ, and activation of the ERK pathway to promote the release of sAppa, can be used to treat Alzheimer's disease. The detailed description above is a detailed description of one of the possible embodiments of the present invention, but the embodiment is not limited to the equivalent implementation or modification of the present invention. It is included in the patent scope of this case. • Ding, mentioned above, this case is not only married in the method of innovation, but also can improve the above-mentioned multi-functions compared with the customary items, and should have fully complied with the novelty and progressiveness of the statutory inventions, and proposes a kiss according to law.恳$You approved the invention request for this invention, in order to invent the invention, to the sense of virtue. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 2 is a flow chart for preparing a 1,2,3,4-tetrahydroheterochemical derivative in the embodiment; FIG. 2 is a (3), 'tetrahydroisotopic method in the second embodiment. Yanlin derivatives inhibit γ-type secretase activity analysis; Figure 2 shows the cytotoxicity test of (2) fine hydrogen isoindole derivatives in Example 2; Western blotting results of precursor protein a (sAPPa) release; _8 is the fourth embodiment! , 2, 3, 4 · four «material derivatives affect the soluble starch-like protein - precursor protein a (sAPPot) relative release degree analysis. [Description of main component symbols] 〇No [References] 1. Prozorovski, VB; Pavlova, LV; Suslova, IM; Belozerova, LV; Kokushkina, AV; Sazonova, AV Russ. J. Appl. Chem. 1995, 68, 589 -593. 2. Liao, Y. R; Wang, BJ; Cheng, Η. T.; Kuo, LH; Wolfe, MSJ Biol. Chem. 2004, 279,49523 - 49532. φ 3. Bakshi, P.; Liao, YF; Gao, J.; Ni, J.; Stein, R.; Yeh, LN; Wolfe, MSJ Biomol. Screening 2005,10,1 - 12. 4. Yogev-Falach, M.; Amit, T. Bar-Am, O.; Youdim, Μ. BH FASEB J. 2003, 17,2325 -2327. '5. Bar-Am, O.; Yogev-Falach, M.; Amit, T.; Sagi, Y. Youdim, Μ. BHJ Neurochem. 2004, 89, 1119 - 1125. 6. Kimberly, WT; LaVoie, MJ; Ostaszewski, BL; Ye, W.; Wolfe, MS; Selkoe, DJ Proc. Natl. Acad. Sci. USA 2003,100,6382 - 6387. 41

Claims (1)

200946119 X. Patent application scope: 1. A 1,2,3,4-tetrahydroisoquinoline derivative having the structure of formula (I): ® R1 : Has the structure of the formula (Π): m is Μ ; or R1 has the structure of formula (HI): η is Μ; wherein: R3X V (IV) ethyl (ch2ch3) or propyl (_(ch2)2CH3) having the formula (IV); wherein R3 is selected from methyl (_CH3) 200946119 wherein R4 is Selected from methyl (-CH3), ethyl (CH2CH3) or propidium CH2) 2CH3); Wherein X is selected from the group consisting of -CHr, oxygen or sulfur; or R2 has the structure of formula (VI): Wherein Y is selected from the group consisting of -CH2-, oxygen or sulfur. 2. The derivative of claim 1, wherein R1 has the structure of formula (8), and R2 has the structure of formula (IV), and R4 is methyl (_CH3). 3. For the towel, please refer to the derivative mentioned in Item 2, which has R1 as propargyl ' and R2 is Ν·ethyl·Ν·methylamino (9). (4) 7 (n_methyl-N-ethylamine oxime)_N_propynyl heart, ^'tetrahydroisoquinolinium (1 2 3 4: (Ν-Μ~.#——οχγ) 43 1 For example, the derivative described in the first paragraph of Wei Wei, wherein R1 has the structure of formula (11), the claw is 1, and R2 has the structure of formula (v), χ s_CH2_. The derivative according to item 4, which is 7-(2) having R1 is propargyl ' and R2 is 1_pyrrolidinyl (ipyrroUdyi) , 3,4_tetrahydroisoquinoline 4 (7-(l-Pyrrolidinecarbamoyloxy)-N-propargyl-l,2,3,4-tetrahydroisoquinoline) 200946119 6. The derivative of claim 2, wherein R1 has a structure of the formula (8), m is 1, and 112 has a structure of the formula (VI), and γ is -CHr. 7. A derivative according to claim 6 which has R1 is propyne Propargyl 'and R2 is i_砒派基基(1) Post-sinking state: (1) Oxygen) N propyl - + fast-yl 2,3,4-tetrahydroisoquinoline (7_ (1 _Piperidinecarbam〇yl〇xy) _N-pr〇pargyl-1,2,3,4-tetrahydroisoquinoline).. 8. The derivative according to claim 1, wherein R1 has a structure of the formula (π), m ® is 1, and 112 has a structure of the formula (W), and Y is oxygen. 9. The derivative according to claim 8 of the patent application, which is 7-(1-overlined) having Ri as propargyl ' and R2 is 1-ophelyl (i_M〇rpholinyl) Oxygen)_n_ propyl group · 1,2,3,4-tetrahydroisophthaleline (7-(1 -Morpholinecarbamoyloxy)-N-propargyl-1,2,3,4-tetrahydroisoquinoline )0 10. Apply for patent The derivative according to the above item 1, wherein R1 has a structure of the formula (m), η ❹ is b and R 2 has a structure of the formula (IV), and R 3 and R 4 are each a methyl group (_CH 3 ). 11. The derivative of claim 10, which has R1 as cyclopropyimethyl' and R2 is N-ethyl-N-methylamine (N-Ethyl-N- Methylamino) '7-(N-Methyl-N-ethylaminemethyl oxime)-N-cyclopropanone methyl-i,2,3,4-tetrahydroisophthalocyanine (7-(N-Methyl) -N-ethylcarbamoyloxy)-N-cyclopropylmethyl-l, 2,3,4-tetrahydr oisoquinoline). 12. The derivative of claim 1, wherein R1 has the structure of formula (羾), n 200946119 is b and R 2 has the structure of formula (V), and X is -CH2-. 13. The derivative according to claim 12, which is a cyclopropylmethyl group wherein R1 is a cyclopropylmethyl group, and R2 is a 1- to 1-pyrrolidyl 7-(1-anthracene). -N-cyclopropaneindolyl-1,2,3,4-tetrahydroisoquinoline (7-(1 -Pyrrolidinecarbamoyl〇xy)-N-cyclopropylmethyl-1,2,3,4- A derivative according to claim 1, wherein Ri has a structure of the formula (Melon), η is 1 ' and R2 has a structure of the formula (VI), and γ is -CHr . • A derivative according to claim 14 which is a cyclopropylmethyl group wherein rj is cyclopropylmethyl and R 2 is i-piperidinyl. Oxygen)-N-cyclopropanylmethyl+2,3,4-tetrahydroisoquinoline (7-(l-Piperidinecarbamoyloxy)-N-cyclopropylmethyl-l,2,3,4-tetrahydroisoq uinoline) 〇16. The derivative of claim 1, wherein R1 has a structure of formula (羾), wherein n is 1' and R2 has a structure of formula (VI), and γ is oxygen. ❿ 17. For the towel, please refer to the touch object described in Item 16 (4), which is 7-(1-?) where R1 is cyclopropylmethyl' and R2 is 1-?M〇rpholinyl.弗琳. 18_- A method for preparing a derivative as described in the patent side of the patent, comprising the following steps: Step (1,2,3,4-tetrahydroisophthalocyanine and trifluoroacetic acid and hydrogen hydroxide unloading) The reaction is carried out to obtain 1^-trifluoroethyl aryl-1,2,3,4-tetrahydroisoquinoline; 45 200946119 Step (b): N-trifluoroethyl fluorenyl-1, 2, 3, 4-tetrahydroisoquinoline is reacted with gas oxime gas and aluminum hydride to obtain 7-gas acetamidine _N_trifluoroethenyl-1,2,3,4-tetrahydroisoquineline; (c): 7-Acetone-N-trifluoroacetamido-U,3,4-tetrahydroisoquinoline is reacted with 3_aperoxybenzoic acid to obtain 7-chloroethyloxy group. -N-trifluoroethylidene-l,2,3,4-tetrahydroisoquinoline; Step (d): 7-Chloroethoxycarbonyl-N-trifluoroethylidene-1,2,3 , 4-tetrahydroisoquineline reacts with sodium thiomethoxide, 7_Hydroxy-1,2,3,4·tetrahydroisoquinoline hydrochloride; _ Step (e): 7-hydroxy-1,2,3,4-tetrahydroisoquinoline hydrochloride Reacting with propynyl bromide or (dimetho) cyclopropane, and potassium carbonate, respectively, can obtain 7-hydroxy-N-propanyl-1,2,3,4-tetrahydroisophthalide or 7- '^-cyclopropanone methyl-1,2,3,4-tetrahydroisoindole|: porphyrin; and step (f): 7-hydroxy_N_propynyl_1,2,3,4 _tetrahydroisoquinoline or 7-hydroxy_^_cyclopropanemethyl-1,2,3,4-tetrahydroisoquinolin' reacts with R2-CO~Cl, and sodium hydride, respectively, 垓R2 is N -ethyl-N-methylamino, 1-oxaridinyl, 1-phenylpyridyl or phoryl, to obtain 1, 2, 3, 4 - 4 having the structure of formula (I) Hydrogen isoquinoline derivative. 19. A pharmaceutical composition comprising the 1,2,3,4-tetrahydroisoindolin derivative or pharmaceutically acceptable salt type as described in claim 1 of the patent application, and Suitable pharmaceutical acceptable excipients or carriers. 20· The pharmaceutical composition according to the scope of patent application 帛W, is used to inhibit the activity of sputum secretase. 21. If the application is in the 19th Material Extracellular activation New adjustment path kinase (extracellular signal-regulated protein kinase, ERK), the further promotion of the release 46200946119 soluble amyloid precursor protein of α. 22. The pharmaceutical composition of claim 19, which is the same as the coffee, to inhibit the activity of the sputum-type secretase and the activation, and the path of the exceriiuiar signai_regUiated pr〇tein kmase 'ERK. Promotes the release of soluble starch-like precursor proteins. 23. The pharmaceutical composition of claim 19 of the scope of the patent application is for treating neuronal cell necrosis due to an increase in insoluble precipitates of mycoplasmate-protein, resulting in loss of memory and learning function, dementia and its cognitive processes Impaired disease. 24. The pharmaceutical composition according to claim 23, wherein the disease in which the cognitive process is impaired is Alzheimer's disease or other dementia or Parkinson&apos;s Disease. 25. The pharmaceutical composition of claim 19, wherein the excipient can be a diluent, a filler, a binder, a disintegrant, a lubricant, and the like. 26. The pharmaceutical composition according to claim 19, wherein the excipient may be microcrystalline cellulose, polyethylene syrup, polyvinylpyrrolidone (PVP), corn house powder, and modified temple. Modified starches, sodium starch glycolate, resin, gelatinized starches, sugars, polyethylene glycol (PEG), polyvinyl alcohol, propylene fiber (hydroxypropylcellulose), methylcellulose, hydroxymethyl cellulose, hydroxypropyl methylcellulose, and the like. 47