CN109912693A - Seven ring aldehyde of RGDS modification, synthesis, anti-thrombus activity and application - Google Patents

Abstract

The invention discloses seven ring aldehyde of the Arg-Gly-Asp-Ser of following formula modification, i.e. (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylene-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone, disclose its preparation method, its anti-arterial thrombus activity is disclosed, thus the invention discloses it to prepare the application in anti-arterial thrombus drug.

Description

Seven ring aldehyde of RGDS modification, synthesis, anti-thrombus activity and application

Technical field

The present invention relates to (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylene-Arg-Gly-Asp- Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone, it is related to its preparation method, is related to its Anti- arterial thrombus activity, thus the present invention relates to it to prepare the application in anti-arterial thrombus drug.The invention belongs to biologies to cure Medicine field.

Background technique

Thrombosis is ischemic heart disease, the common pathology of ishemic stroke and phlebothrombosis.In the world, it lacks Death toll caused by hemorrhagic heart disease and ishemic stroke accounts for the 1/4 of whole Died Of Disease numbers.And phlebothrombosis is then not Developed country, medium-developed country and the highly developed main Disease Spectrum of country.Thrombus can make a series of relevant diseases Disease deteriorates, such as the blocking thrombus of the biomaterial pipe seldom occurred can be to change pipe repeatedly, or receive thromboembolism treatment or length The patient that phase receives anticoagulant therapy, which brings, is difficult to expect consequence, can cause to receive percutaneous coronary intervention treatment future trouble Person's embolism again, can with heparin-induced thrombocytopenia shape, and curved coronary artery thrombosis can cause it is acute Coronary syndrome.In addition, thrombosis is the complication of related disease, for example, bulk cerebral venous thrombosis with epileptics The complication of early stage pregnant woman, and stent thrombosis are the serious complication of percutaneous coronary intervention treatment patient.It can See, invents novel antithrombotic reagent with clinical importance.

B-carboline is the important pharmacophore for inhibiting thrombus.However B-carboline, such as 3S-1,2,3,4- tetrahydro-beta-carboline -3- Carboxylic acid will obtain anti-arterial thrombus purpose, not only need to be injected intravenously, and dosage needs 5 μm of ol/kg.From 3S-1,2,3,4- From the point of view of tetrahydro-β-anti-arterial thrombus of carboline -3- carboxylic acid, both expected to change into intravenous injection it is oral, and expect depressant Amount.Inventor assumes that two B-carboline pharmacophores merge, such as two (1R, 3S) -1- carbonyl methyl -2,3,4,9- tetrahydro -1H- Pyrido [3,4-b] indole -3-carboxylic acid's intermolecular condensation is (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- carbonyls Methyl) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl -1 ', 4 '-diketone Ying Youqiang oral antithrombotic acitivity.Hair Bright people it is further assumed that, toward this novel seven rings aldehyde aldehyde radical connection Arg-Gly-Asp-Ser should have stronger oral antithrombotic Activity.According to this hypothesis, the present invention is inventors herein proposed.

Summary of the invention

First content of the invention is to provide (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] of following formula and bis- { (1R- Methylene-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone.

Second content of the invention is to provide (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylene - Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyls -1 ', 4 '-diketone preparation method, should Method includes:

1) L-Trp benzyl ester is subjected to Pictet- with 1,1,3,3- tetramethoxy propane under trifluoroacetic catalysis Spengler condensation, obtains (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- benzyl carboxylate Ester (1)；

2) (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- benzyl carboxylate in methyl alcohol Ester hydrogenolysis under Pd/C catalysis takes off benzyl and obtains (1R, 3S) -1- (dimethoxy ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- Carboxylic acid (2)；

3) in the presence of benzotriazole-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HBTu), (1R, 3S) -1- (two - 2 base of methoxyethyl) -2,3,4,9- tetrahydro-beta-carboline -3- carboxylic acid is intermolecular in anhydrous DMF (N,N-dimethylformamide) Condensation is (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- dimethoxy ethyl -2- base) -2,3,4,9- tetrahydros - 1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (3)；

4) use glacial acetic acid for solvent, concentrated hydrochloric acid and water be catalyst (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] simultaneously The conversion of bis- { (1R- dimethoxy ethyl -2- base) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyls } -1 ', 4 '-diketone (3) For (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- carbonyl methyl) -2,3,4,9- tetrahydro -1H- pyridines [3,4-b] Diindyl } -1 ', 4 '-diketone (4)；

5) use dicyclohexylcarbodiimide for condensing agent, I-hydroxybenzotriazole is the method for the liquid phase condensations of catalyst Synthesize HClArg (NO₂)-Gly-Asp(OBzl)-Ser-OBzl；

It 6) is reducing agent by HClArg (NO with sodium cyanoborohydride₂)-Gly-Asp (OBzl)-Ser-OBzl is connected to (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- carbonyl methyl) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] and Yin Diindyl } -1 ', 4 '-diketone (4) aldehyde radical on, synthesis (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { [1R- methylene-Arg (NO₂)-Gly-Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl -1 ', 4 '-diketone (5)；

7) simultaneously double in trifluoracetic acid and in the mixed solvent (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] of trifluoromethanesulfonic acid { [1R- methylene-Arg (NO₂)-Gly-Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] and Yin Diindyl } -1 ', 4 '-diketone (5) remove nitro and benzyl ester obtains (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (Asias 1R- First-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (6).

Third content of the invention is evaluation (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylenes- Arg- Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl -1 ', 4 '-diketone anti-arterial thrombus make With.

Detailed description of the invention

Fig. 1 (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and it is bis- (1R- methylene-Arg-Gly-Asp-Ser) -2,3, 4,9- tetrahydro -1H- pyridine [3,4-b] diindyls } -1 ', 4 '-diketone synthetic route (i) methylene chloride, trifluoracetic acid； (ii)CH₃OH, Pd/C, H₂；(iii) HBTu, NMM, anhydrous DMF；(iv)H₂O, glacial acetic acid, concentrated hydrochloric acid；(v) DCC, HOBt, NMM, THF；(vi) ethyl acetate solution (4M) of hydrogen chloride；(vii)CH₃OH, NaOH solution (2M)；(viii) methylene chloride, NaCNBH₃, anhydrous MgSO₄；(ix) trifluoracetic acid, trifluoromethanesulfonic acid.

Specific embodiment

In order to which the present invention is further explained, a series of embodiments are given below.These embodiments be entirely it is illustrative, it Only be used to the present invention is specifically described, be not construed as limitation of the present invention.

Embodiment 1 prepares (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- benzyl carboxylate Ester (1)

180mL methylene chloride under ice bath, 10mL1,1,3,3- tetramethoxy propane and the trifluoroacetic mixed solution of 10mL Stir 40min.Later, 10.25g (34.86mmol) L-Trp-OBzl is added afterwards.14h, TLC is stirred at room temperature in reaction mixture (petrol ether/ethyl acetate=1:1) shows that L-Trp-OBzl disappears.Reaction solution is respectively with saturation NaHCO₃Aqueous solution washes (40mL × 3), saturation NaCl aqueous solution is washed (40mL × 3), isolated methylene chloride anhydrous Na SO₄Dry 12h, filtering, filtrate decompression Concentration, residue are purified (petrol ether/ethyl acetate=3:1) with silica gel column chromatography, obtain 5.12g (37%) title compound Object is brownish red grease.ESI-MS(m/e):393[M+H]^-。

Embodiment 2 prepares (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- carboxylic acid (2)

To 5.12g (13.00mmol) (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline - 500mg Pd/C is added in the solution of 3- benzyl carboxylate and 100mL methanol, is filled with hydrogen and 18h is stirred at room temperature, TLC (petroleum ether/ Ethyl acetate=1:1) show that 1 disappears, terminate reaction.Pd/C is filtered out, filtrate decompression is concentrated to dryness, and obtains 3.63g (92%) title Compound is glassy yellow grease.ESI-MS(m/e):303[M-H]^-。

Embodiment 3 prepare (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and it is bis- (1R- dimethoxy ethyl -2- base) -2, 3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (3)

To 6.12g (20.13mmol) (1R, 3S) -1- (2,2- dimethoxy-ethyl -2- base) -2,3,4 under ice bath stirring, 3.00g (22.22mmol) HBTu is added in 9- tetrahydro-β-carboline -3- carboxylic acid (2) and the solution of 100mL anhydrous DMF.Then it uses The pH of reaction solution is adjusted to 8-9 by N-methyl morpholine, and point TLC (petrol ether/ethyl acetate=1:1) display 2 disappears afterwards for 24 hours for reaction It loses, terminates reaction.Reaction solution is concentrated under reduced pressure, and residue 150mL ethyl acetate dissolves.Later, respectively with saturation NaHCO₃Water Solution is washed (30mL × 3), and saturation NaCl aqueous solution washes (30mL × 3), 5%KHSO₄Aqueous solution is washed (3 0mL × 3), and NaCl is saturated Aqueous solution is washed (30mL × 3), and NaHCO is saturated₃Aqueous solution is washed (30mL × 3), and saturation Na Cl aqueous solution washes (30mL × 3).Acetic acid The methacrylate layer dry 12h of anhydrous sodium sulfate, filtering, filtrate decompression concentration.Obtained dark yellow solid is separated with silica gel column chromatography (petrol ether/ethyl acetate=2:1) obtains 1.14g (10%) title compound, is yellow solid.ESI-MS(m/e):573[M+ H]⁺；¹HNMR(300MHz,DMSO-d₆) δ/ppm=11.08 (s, 2H), 7.39 (d, J=7.5Hz, 2H), 7.34 (d, J= 8.1Hz, 2H), 7.08 (t, J=7.2Hz, 2H), 6.97 (t, J=7.2Hz, 2H), 5.93 (dd, J₁=3.6Hz, J₂= 9.0Hz,2H),4.62(dd,J₁=3.3Hz, J₂=11.4Hz, 2H), 4.53 (t, J=4.8Hz, 2H), 3.34 (s, 6H), 3.26(s,6H),3.20(m,2H),2.79 (m,2H),2.24(m,4H)。

Embodiment 4 prepares (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- carbonyl methyl) -2,3,4,9- four Hydrogen -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (4)

To 380mg (0.66mmol) (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- bis- under ice bath and stirring Methoxyethyl -2- base) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (3) and 9mL glacial acetic acid 6mL water and 3mL concentrated hydrochloric acid are added in solution, stirs 2h, TLC shows that (petrol ether/ethyl acetate=1:1) 3 disappears, and terminates anti- It answers.Under ice bath, reacting liquid pH value is adjusted to 7 with NaOH aqueous solution (2M).Obtained solution be extracted with ethyl acetate (50 mL × 3), combined ethyl acetate layer is respectively with saturation NaHCO₃Aqueous solution is washed (30mL × 3), saturation NaCl aqueous solution wash (30mL × 3), 12h is dried with anhydrous sodium sulfate in ethyl acetate phase, filters off sodium sulphate, and filtrate decompression is concentrated to get 288mg (90%) Title compound is yellow solid.ESI-MS(m/e):481[M+H]⁺；¹H-NMR(300MHz, DMSO-d₆) δ/ppm= 10.95 (s, 2H), 9.79 (m, 2H), 7.42 (d, J=8.1Hz, 2H), 7.38 (d, J=7.5Hz, 2H), 7.10 (t, J= 6.9Hz, 2H), 6.99 (t, J=6.9Hz, 2H), 6.22 (dd, J₁=3.6Hz, J₂=9.0Hz, 2H), 4.67 (dd, J₁= 3.3Hz,J₂=11.4Hz, 2H), 3.26 (m, 2H), 3.11 (m, 4H), 2.84 (m, 2H).

Embodiment 5 prepares Boc-Arg (NO₂)-Gly-OBzl

To 3.19g (10.00mmol) Boc-Arg (NO under ice bath₂) be added in the solution of 150mL anhydrous tetrahydro furan 1.35g (10.00mmol) HOBt is added 2.27g (11.00mmol) DCC after stirring 5min, stirs 30min under ice bath, obtains anti- Answer liquid A.Under ice bath, 3.50g (12.00mmol) HClGly-OBzl is added in reaction solution A, adjusts pH with N- methyl morpholine It is worth 9.Anti- 20h is stirred at room temperature in reaction mixture, and TLC (methylene chloride/methanol=20:1) shows that Boc-Arg (NO2) disappears. Filter out the insoluble matter in reaction solution, filtrate decompression concentration, residue 150mL ethyl acetate dissolves, and filters off insoluble colourless solid Body, filtrate is respectively with saturation NaHCO₃Aqueous solution is washed (30mL × 3), and saturation NaCl aqueous solution washes (30mL × 3), 5%KHSO₄Water Solution is washed (30mL × 3), and saturation NaCl aqueous solution is washed (30mL × 3), is saturated NaHCO₃Aqueous solution is washed (30mL × 3), saturation NaCl aqueous solution washes (30mL × 3), and the obtained ethyl acetate phase dry 12h of anhydrous sodium sulfate filters off sodium sulphate, filtrate decompression Concentration obtains yellow syrup, and the dissolution of 30mL methylene chloride is added, is placed in 4 DEG C of standing 12h；White solid is precipitated, is filtered under diminished pressure, 4.32g (93%) title compound is obtained, is colorless solid.ESI-MS(m/e):467[M+H]⁺。

Embodiment 6 prepares Boc-Arg (NO₂)-Gly

To 4.32g (9.25mmol) Boc-Arg (NO under ice bath₂)-Gly-OBzl and 30mL methanol solution in 2M is added dropwise NaOH solution, adjusting solution ph are 13-14.Reaction mixture ice bath stirring anti-1h, TLC (methylene chloride/methanol, 20/1, V/v contains 2 drops/acetic acid) display Boc-Arg (NO₂)-Gly-OBzl disappearance.Saturation KHSO is added dropwise under ice bath₄Solution is adjusted molten The pH of liquid is 7, and decompression rotation removes methanol.Saturation KHSO is added dropwise in residue under ice bath₄Solution, the pH for adjusting solution is 2-3, uses second Acetoacetic ester extracts (50mL × 3), combined ethyl acetate layer, washes (30mL × 3) with saturation NaCl aqueous solution, obtained acetic acid second Ester mutually with the dry 12h of anhydrous sodium sulfate, filters off sodium sulphate, filtrate decompression concentration, and 3.21 g (92%) title compound is colourless Syrup.

Embodiment 7 prepares Boc-Asp (OBzl)-Ser-OBzl

According to the method for embodiment 5 from 3.23g (10.00mmol) Boc-Asp (OBzl) and 2.78g (12.00mmol) HClSer-OBzl obtains 4.43g (89%) title compound, is yellow syrup.ESI-MS(m/e):501[M+H]⁺.

Embodiment 8 prepares HClAsp (OBzl)-Ser-OBzl

By 4.43g (8.86mmol) Boc-Asp (OBzl)-Ser-OBzl with 20mL anhydrous ethyl acetate dissolve, then in The ethyl acetate solution (4M) of 50mL hydrogen chloride is added under ice bath into solution, is stirred at room temperature 6h, TLC (methylene chloride/methanol/ Acetic acid, 20/1/2 drop, v/v/v) display Boc-Asp (OBzl)-Ser-OBzl disappearance.Reaction solution is concentrated under reduced pressure, residue is used Anhydrous ethyl acetate is concentrated under reduced pressure again after redissolving, then is redissolved with anhydrous ethyl acetate.The operation is at least repeated 3 times.Final residual Object is washed with anhydrous ether, obtains 3.58g (93%) title compound, is yellow solid.

Embodiment 9 prepares Boc-Arg (NO2)-Gly-Asp (OBzl)-Ser-OBzl

According to the method for embodiment 5 from 2.50g (6.65mmol) Boc-Arg (NO2)-Gly and 3.48g (7.97mmol) HClAsp (OBzl)-Ser-OBzl obtains 2.55g (42%) title compound, is colourless powder.ESI-MS(m/e):759 [M+H]⁺。

Embodiment 10 prepares HClArg (NO2)-Gly-Asp (OBzl)-Ser-OBzl

According to the method for embodiment 8 from 843mg (1.11mmol) Boc-Arg (NO2)-Gly-Asp (OBzl)-Ser-OBzl 729mg (94%) title compound is obtained, is colorless solid.

Embodiment 11 prepares (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { [1R- methylene-Arg (NO₂)- Gly- Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (5)

By 168mg (0.35mmol) (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and it is bis- (1R- carbonyl methyl) -2,3,4, 9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (4) are dissolved in 15mL methylene chloride, with triethylamine tune under ice bath PH value is saved to 9.By 729mg (1.05mmol) HClArg (NO₂)-Gly-Asp (OBzl)-Ser-OBzl 15mL methylene chloride It dissolves, pH value is adjusted to 9 with triethylamine under ice bath, is added drop-wise under ice bath in the dichloromethane solution of compound 4, adds 100mg MgSO₄.After reaction mixture ice bath stirring 1h, 27mg NaCNBH is added every 1h₃, 108mg NaCNBH is added altogether₃.Later, TLC (methylene chloride/methanol=7:1) for 24 hours is stirred at room temperature and shows that compound 4 disappears, terminates reaction.Reaction solution is respectively with saturation NaHCO₃Aqueous solution is washed (7mL × 3), and saturation NaCl aqueous solution washes (7mL × 3), and obtained methylene chloride mutually uses anhydrous sodium sulfate After dry 12h, sodium sulphate, filtrate decompression concentration are filtered off, obtained yellow solid silica gel column chromatography purifies (methylene chloride/first Alcohol=20:1), 52mg (8%) title compound is obtained, is faint yellow solid.ESI-MS(m/e):1765[M+H]⁺；¹H-NMR (300MHz,DMSO-d₆) δ/ppm=11.00 (s, 2H), 8.58 (m, 2H), 8.37 (m, 6H), 7.38~7.30 (m, 24H), 7.07 (t, J=7.8Hz, 2H), 6.96 (t, J=7.5Hz, 2H), 5.93 (m, 2H), 5.12 (m, 8H), 4.80 (m, 2H), 4.57(dd,J₁=3.9Hz, J₂=11.4Hz, 2H), 4.48 (dd, J₁=4.5Hz, J₂=8.4Hz, 2H), 3.83 (m, 4H), 3.66 (m, 2H), 3.59 (m, 4H), 3.25 (m, 2H), 3.19 (m, 10H), 2.77~2.51 (m, 10H), 2.27 (m, 2H), 2.06(m,4H),1.62(m,8H)。

Embodiment 12 prepares (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { [1R- methylene-Arg-Gly-Asp- Ser] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (6)

By 50mg (0.03mmol) (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { [1R- methylene-Arg under ice bath (NO₂)-Gly-Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl -1 ', 4 '-diketone (5) it is dissolved with 1.5mL trifluoroacetic acid.Later, 0.5mL trifluoromethanesulfonic acid is added, stirs 30min, TLC (dichloromethane under ice bath Alkane/methanol=7:1) the display disappearance of compound 5.Add 25mL anhydrous ether under ice bath into reaction mixture, stirs 10min, it is quiet It sets, incline supernatant.Residue adds 25mL anhydrous ether again, stirs 10min, stands, incline supernatant.The operation at least repeats 3 times.Obtained 5% acetate dissolution of yellow black solid 1mL, solution are purified with Sephadex, and it is titled to obtain 6mg (16%) Object is closed, is faint yellow solid.ESI-MS(m/e):1315[M+H]⁺；¹H-NMR(300MHz, DMSO-d₆) δ/ppm=12.56 (s,4H),10.77(s,2H),9.14(m,2H),8.95(m,2H),8.48(m,2H), 8.06(m,2H),7.44(m,8H), 7.10 (t, J=7.8Hz, 2H), 7.03 (t, J=7.2Hz, 2H), 5.82 (m, 2H), 4.83~4.55 (m, 6H), 3.96 (m, 4H), 3.87 (m, 2H), 3.63 (m, 4H), 3.27 (m, 2H), 3.14 (m, 10H), 2.87~2.63 (m, 10H), 2.35 (m, 2H),2.23(m,2H),1.79(m,4H),1.55(m,4H)。

Embodiment 13 evaluates (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylene-Arg-Gly-Asp- Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (6) anti-arterial thrombus effect

Experimental material: aspirin (Aspirin, CAS:50-78-2, article No.: 298969, the limited public affairs of lark prestige science and technology Department), urethane (CAS:51-79-6, article No.: 30191228, Sinopharm Chemical Reagent Co., Ltd.), heparin sodium (CAS: 9041-08-1, article No.: 542858, lark prestige Science and Technology Ltd.), physiological saline (Shijiazhuang Siyao Co., Ltd), anhydrous second Ether (Beijing chemical reagents corporation), silicone oil (Beijing chemical reagents corporation), two citric acid monohydrate trisodiums (CMCNa, article No.: 20170713, Beijing Chemical Plant).

Experimental animal: SD strain rats, male, 200 ± 20g tie up the limited public affairs of tonneau China experimental animal technology purchased from Beijing Department.

Experimental method: experiment bypasses tuft method using arteria carotis communis-vena jugularis externa extracorporal circulatory system.

Grouping and dosage: (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- methylene-Arg-Gly-Asp- Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (abbreviation compound 6) dosage be 0.1 μ Mol/kg, the dosage of positive control aspirin are 167 μm of ol/kg, negative control 0.5%CMCNa.

The dosage form of reagent: anesthetic is the normal saline solution (20%) of urethane, and anti-coagulants is the physiology salt of heparin sodium Aqueous solution (42mg/100mL), compound 6 are the suspension of 0.5%CMCNa, and aspirin is the suspension of 0.5%CMCNa.

Bypass intubation: the bypass intubation of right jugular vein and left neck artery is 1.0mm, outer diameter 2.0mm, length 10.0 by internal diameter The polyethylene pipe of cm is constituted.The tubule pulled into as angle is heated in their one end, to be inserted into right jugular vein and left neck artery. They constitute the both ends of bypass intubation.It is 3.5mm that the middle section being intubated, which is bypassed, by internal diameter, and the polyethylene pipe of a length of 8.0cm is constituted.

Thrombus load line: a length of 6.0cm, weight are the crude silk thread in surface of 4.0 ± 0.1mg.

Bypass intubation assembling: three sections of Interior Wall of Polythene Pipe use diethyl ether solution (1%) silanization of silicone oil, dry completely Afterwards, silk thread is placed in the polyethylene pipe of middle section in arteria carotis intubation direction, three sections of polyethylene pipes is assembled and consolidated using sealed membrane It is fixed, heparin need to be full of before intubation in pipe.

Experimental implementation: according to the dosage of 0.1 μm of ol/kg by the suspension of the CMCNa of compound 6 with 0.3mL/100g weight To rat oral gavage, the suspension of the CMCNa of aspirin is given greatly with 0.3mL/100g weight according to the dosage of 167 μm of ol/kg Mouse stomach-filling, and by CMCNa with 0.3mL/100g weight to rat oral gavage.The urethane solution of intraperitoneal injection 20% after 30min Anaesthetized (0.7mL/100g).Rat is fixed on plate by dorsal position, cuts off skin of neck, separates right common carotid artery and left neck Outer vein respectively ligatures the distal end of right common carotid artery and left vena jugularis externa with surgical thread, in quiet outside exposed left neck Arteries and veins cuts a V-shape osculum, and the vein end angle of the bypass intubation made above is inserted into the proximal part of left vena jugularis externa opening, is inserted Blood vessel and polyethylene pipe are fixed with surgical thread at pipe.Pass through the accurate note of bypass intubation according to the dosage of 0.1mL/100g weight Enter heparin sodium water solution, syringe does not withdraw polyethylene pipe.Right common carotid artery proximal part is clamped with artery clamp, in exposed artery On cut a V-shape osculum, the tip of polyethylene pipe is removed from syringe, the proximal part of right common carotid artery is inserted a tube into, uses hand Art line fixes arteries and polyethylene pipe, unclamps artery clamp, establishes extracorporal circulatory system bypass.Rat temperature and bypass is maintained to insert Blood flow is unobstructed in pipe, and vein end is first intubated to cut whether observation blood circulation is smooth after extracorporal circulatory system 15min, from moving for intubation Arteries and veins end removal of thromboses line weighs and records thrombus weight after sucking the blood on silk thread on filter paper, represent anti-arterial thrombus activity.Number It is counted according to using t inspection.Experimental data is shown in Table 1.

Table 1 statistics indicate that, (2 ' S, 5 ' the S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and double under 0.1 μm of ol/kg oral dose (the letter of { (1R- methylene-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone Claim compound 6) it can conspicuousness inhibition rat formation thrombus.This effective dose is intravenous injection 3S-1,2,3,4- tetrahydro-β-click The 1/50 of 5 μm of ol/kg of effective dose of quinoline -3- carboxylic acid antithrombotic.The compound of the present invention 6 is not only by 3S-1,2,3,4- tetrahydro- The intravenous injection of B-carboline -3- carboxylic acid is changed into oral, and reduces dosage.As it can be seen that the present invention has unexpected technology Effect.

The anti-arterial thrombus activity of 1 compound 6 of table

Compound	Dosage	Thrombus weight (mean value ± SD mg)
			0.5%CMCNa	3mL/kg	29.29±4.94
Aspirin	167μmol/kg	18.98±4.24a
			Compound 6	0.1μmol/kg	21.08±3.09a

And 0.5%CMCNa ratio P < 0.01 a)；N=8.

Claims (3)

1. the Arg-Gly-Asp-Ser of following formula modifies seven ring aldehyde, i.e. (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] is simultaneously double { (1R- methylene-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone,

2. the Arg-Gly-Asp-Ser of claim 1 modifies the preparation method of seven ring aldehyde, this method comprises:

1) L-Trp benzyl ester is subjected to Pictet- with 1,1,3,3- tetramethoxy propane under trifluoroacetic catalysis Spengler condensation, obtains (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- benzyl carboxylate (1)；

2) (1R, 3S) -1- (dimethoxy-ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- benzyl carboxylate exists in methyl alcohol Pd/C is catalyzed the de- benzyl of lower hydrogenolysis and obtains (1R, 3S) -1- (dimethoxy ethyl -2- base) -2,3,4,9- tetrahydro-beta-carboline -3- carboxylic acid (2)；

3) in the presence of benzotriazole-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HBTu), (1R, 3S) -1- (dimethoxy - 2 base of ethyl) -2,3,4,9- tetrahydro-beta-carboline -3- carboxylic acid intermolecular condensation in anhydrous DMF (N,N-dimethylformamide) is (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- dimethoxy ethyl -2- base) -2,3,4,9- tetrahydro -1H- pyridines [3,4-b] diindyl } -1 ', 4 '-diketone (3)；

4) use glacial acetic acid for solvent, concentrated hydrochloric acid and water are that catalyst (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] is simultaneously double { (1R- dimethoxy ethyl -2- base) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (3) are converted into (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (1R- carbonyl methyl) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] and Yin Diindyl } -1 ', 4 '-diketone (4)；

5) use dicyclohexylcarbodiimide for condensing agent, I-hydroxybenzotriazole is the method synthesis of the liquid phase condensations of catalyst HCl·Arg(NO₂)-Gly-Asp(OBzl)-Ser-OBzl；

It 6) is reducing agent by HClArg (NO with sodium cyanoborohydride₂)-Gly-Asp (OBzl)-Ser-OBzl be connected to (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] is simultaneously bis- { (1R- carbonyl methyl) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyls } - On the aldehyde radical of 1 ', 4 '-diketone (4), synthesis (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { [1R- methylene-Arg (NO₂)-Gly-Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl -1 ', 4 '-diketone (5)；

7) simultaneously double in trifluoracetic acid and in the mixed solvent (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] of trifluoromethanesulfonic acid { [1R- methylene-Arg (NO₂)-Gly-Asp (OBzl)-Ser-OBzl] -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] and Yin Diindyl } -1 ', 4 '-diketone (5) remove nitro and benzyl ester obtains (2 ' S, 5 ' S)-tetrahydro pyrazine [1 ', 2 ': 1,6] and bis- { (Asias 1R- First-Arg-Gly-Asp-Ser) -2,3,4,9- tetrahydro -1H- pyridine [3,4-b] diindyl } -1 ', 4 '-diketone (6).

3. the Arg-Gly-Asp-Ser of claim 1 modifies seven ring aldehyde and is preparing the application in anti-arterial thrombus drug.