CN110563799A - RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof - Google Patents

RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof Download PDF

Info

Publication number
CN110563799A
CN110563799A CN201810561696.3A CN201810561696A CN110563799A CN 110563799 A CN110563799 A CN 110563799A CN 201810561696 A CN201810561696 A CN 201810561696A CN 110563799 A CN110563799 A CN 110563799A
Authority
CN
China
Prior art keywords
tetrahydro
arg
bis
gly
asp
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201810561696.3A
Other languages
Chinese (zh)
Other versions
CN110563799B (en
Inventor
赵明
冯琦琦
桂琳
彭师奇
石林峄
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Capital Medical University
Original Assignee
Capital Medical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Capital Medical University filed Critical Capital Medical University
Priority to CN201810561696.3A priority Critical patent/CN110563799B/en
Publication of CN110563799A publication Critical patent/CN110563799A/en
Application granted granted Critical
Publication of CN110563799B publication Critical patent/CN110563799B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Hematology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Diabetes (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The invention discloses Arg-Gly-Asp-Ser modified heptacyclic aldehyde, namely (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1, 6)]And bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]The invention discloses a method for preparing the indole-1 ', 4' -diketone and the anti-venous thrombosis activity thereof, thus the invention discloses the application of the indole-1 ', 4' -diketone in preparing the anti-venous thrombosis medicine.

Description

RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
Technical Field
the invention relates to (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indole } -1 ', 4' -dione, to a process for its preparation, to its anti-venous thrombosis activity and thus to its use in the preparation of anti-venous thrombosis medicaments. The invention belongs to the field of biological medicine.
Background
Both arterial and venous thrombosis have become diseases with high morbidity and mortality, and the worldwide number of deaths from ischemic heart disease and ischemic stroke is 1/4 of all deaths due to disease. Venous thrombosis is a major disease burden in less-developed, moderately-developed and highly-developed countries. Venous thrombosis includes primarily deep vein thrombosis and pulmonary embolism. The number of patients with deep vein thrombosis and pulmonary embolism exceeds the total number of patients with myocardial infarction and apoplexy, and is higher than the total number of deaths caused by breast cancer and AIDS. Since the incidence of venous thrombosis increases exponentially with age, the threat of this condition to the health of people in the aging countries of our country is particularly severe. If the population cardinality is considered, the absolute negative influence on the national civilization of China is particularly serious. Therefore, the prevention and treatment of venous thrombosis have been the focus of attention in the field of medicine, and the invention of novel anti-venous thrombosis medicines has clinical importance.
The beta-carboline is an important pharmacophore for inhibiting thrombus. The inventors hypothesize that the intermolecular condensation of two beta-carboline pharmacophores, e.g., two (R) -1-carbonylmethyl-2, 3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole-3-carboxylic acids, results in (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-carbonylmethyl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione. The novel heptacyclic aldehyde has strong anti-vein thrombosis activity. The inventors further hypothesized that the connection of Arg-Gly-Asp-Ser to the aldehyde group of this novel heptacyclic aldehyde should have greater anti-thrombogenic activity. Based on this assumption, the inventors have proposed the present invention.
Disclosure of Invention
In a first aspect of the invention there is provided (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione of the formula.
In a second aspect of the present invention, there is provided a process for the preparation of (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione, which process comprises:
1) Carrying out Pictet-Spengler condensation on L-tryptophan benzyl ester and 1,1,3, 3-tetramethoxypropane under the catalysis of trifluoroacetic acid to obtain (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-benzyl carboxylate (1);
2) Carrying out hydrogenolysis debenzylation on (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylic acid benzyl ester in methanol under the catalysis of Pd/C to obtain (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylic acid (2);
3) Intermolecular condensation of (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro- β -carboline-3-carboxylic acid in anhydrous DMF (N, N-dimethylformamide) in the presence of benzotriazole-N, N ' -tetramethyluronium hexafluorophosphate (HBTu) to (2 ' S,5 ' S) -tetrahydropyrazino [1 ', 2 ': 1,6] and bis { (1R) - [ 1-dimethoxyethyl-2-yl ] -2,3,4, 9-tetrahydro-1H-pyridin [3,4-b ] oxindole } -1 ', 4 ' -dione (3);
4) Using glacial acetic acid as a solvent, concentrated hydrochloric acid and water as catalysts to convert (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-dimethoxyethyl-2-yl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione (3) into (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-carbonylmethyl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione (4);
5) Liquid phase condensation method for synthesizing HCl & Arg (NO) by using dicyclohexylcarbodiimide as condensing agent and 1-hydroxybenzotriazole as catalyst2)-Gly-Asp(OBzl)-Ser-OBzl；
6) Using sodium cyanoborohydride as a reducing agent to react HCl & Arg (NO)2) -Gly-Asp (OBzl) -Ser-OBzl to (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6]And bis { (1R) - [ 1-Carbonylmethyl group]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]Synthesis of (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] on the aldehyde group of indole } -1 ', 4' -dione (4)]And bis { (1R) - [ 1-ethyl-Arg (NO)2)-Gly-Asp(OBzl)-Ser-OBzl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione (5);
7) (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6] in a mixed solvent of trifluoroacetic acid and trifluoromethanesulfonic acid]And bis { (1R) - [ 1-ethyl-Arg (NO)2)-Gly-Asp(OBzl)-Ser-OBzl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]Removing nitro and benzyl ester from indole } -1 ', 4' -diketone (5) to obtain (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1, 6)]And bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione (6).
The third aspect of the present invention is to evaluate the antithrombotic effect of (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione.
drawings
FIG. 1(2 'S, 5' S) -tetrahydropyrazines [1 ', 2': 1,6]And bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]Synthetic routes to indolino } -1 ', 4' -dione (i) dichloromethane, trifluoroacetic acid; (ii) CH (CH)3OH，Pd/C，H2(ii) a (iii) HBTu, NMM, anhydrous DMF; (iv) h2O, glacial acetic acid and concentrated hydrochloric acid; (v) DCC, HOBt, NMM, THF; (vi) hydrogen chloride in ethyl acetate (4M); (vii) CH (CH)3OH, NaOH solution (2M); (viii) dichloromethane, NaCNBH3Anhydrous MgSO (MgSO)4(ii) a (ix) Trifluoroacetic acid and trifluoromethanesulfonic acid.
Detailed Description
To further illustrate the invention, a series of examples are given below. These examples are purely illustrative and are intended to be a detailed description of the invention only and should not be taken as limiting the invention.
EXAMPLE 1 preparation of benzyl (3S) -1- (2, 2-Dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylate (1)
In ice bath 180mL of dichloromethane, 10mL of 1,1,3, 3-tetramethoxypropane and 10mL of trifluoroacetic acidThe mixed solution was stirred for 40 min. Then, 10.25g (34.86mmol) of L-Trp-OBzl was added. The reaction mixture was stirred at rt for 14h and TLC (petroleum ether/ethyl acetate ═ 1:1) showed disappearance of L-Trp-OBzl. The reaction solution is respectively saturated NaHCO3Aqueous solution (40mL × 3), saturated aqueous NaCl solution (40mL × 3), separated dichloromethane dried over anhydrous sodium sulfate for 12h, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate ═ 3:1) to give 5.12g (37%) of the title compound as a brownish red oil. ESI-MS (M/e) 393[ M + H ]]-。
EXAMPLE 2 preparation of (3S) -1- (2, 2-Dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylic acid (2)
To a solution of 5.12g (13.00mmol) of benzyl (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro- β -carboline-3-carboxylate in 100mL of methanol was added 500mg of Pd/C, charged with hydrogen and stirred at room temperature for 18h, and TLC (petroleum ether/ethyl acetate 1:1) showed 1 disappearance, and the reaction was terminated. Pd/C was filtered off, and the filtrate was concentrated to dryness under reduced pressure to give 3.63g (92%) of the title compound as a bright yellow oil. ESI-MS (M/e):303[ M-H]-。
EXAMPLE 3 preparation of (2 'S, 5' S) -Tetrahydropyrazine [1 ', 2': 1,6] and bis { (1R) - [ 1-dimethoxyethyl-2-yl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione (3)
To a solution of 6.12g (20.13mmol) of (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro- β -carboline-3-carboxylic acid (2) in 100mL of anhydrous DMF was added 3.00g (22.22mmol) of HBTU with stirring in ice bath. The reaction solution was then adjusted to pH 8-9 with N-methylmorpholine, and after 24h the reaction was terminated by point TLC (petroleum ether/ethyl acetate 1:1) indicating 2 had disappeared. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in 150mL of ethyl acetate. Then, saturated NaHCO was used separately3Aqueous solution (30 mL. times.3), saturated aqueous NaCl solution (30 mL. times.3), 5% KHSO4aqueous solution (30 mL. times.3), saturated aqueous NaCl solution (30 mL. times.3), saturated aqueous NaHCO solution3The resulting mixture was washed with an aqueous solution (30 mL. times.3) and with a saturated aqueous NaCl solution (30 mL. times.3). The ethyl acetate layer was dried over anhydrous sodium sulfate for 12 hours, filtered, and the filtrate was concentrated under reduced pressure. The dark yellow solid obtained is chromatographed on a silica gel column (petroleum ether/ethyl acetate 2:1) to give 1.14g (10%) of the title compound as a yellow solid. ESI-MS (M/e):573[ M + H]+；1H NMR(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)。
EXAMPLE 4 preparation of (2 'S, 5' S) -Tetrahydropyrazine [1 ', 2': 1,6] and bis { (1R) - [ 1-Carbonylmethyl ] -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indoline } -1 ', 4' -dione (4)
380mg (0.66mmol) of (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6] are stirred in an ice bath]And bis { (1R) - [ 1-dimethoxyethyl-2-yl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]To a solution of indole } -1 ', 4' -dione (3) and 9mL of glacial acetic acid were added 6mL of water and 3mL of concentrated hydrochloric acid, and stirred for 2 hours, and TLC showed (petroleum ether/ethyl acetate ═ 1:1)3 to disappear, and the reaction was terminated. The reaction was adjusted to pH 7 with aqueous NaOH (2M) in an ice bath. The resulting solution was extracted with ethyl acetate (50 mL. times.3), and the combined ethyl acetate layers were each extracted with saturated NaHCO3Washed with aqueous solution (30 mL. times.3), washed with saturated aqueous NaCl solution (30 mL. times.3), the ethyl acetate phase was dried over anhydrous sodium sulfate for 12h, the sodium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to give 288mg (90%) of the title compound as a 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)。
EXAMPLE 5 preparation of Boc-Arg (NO2) -Gly-OBzl
To a solution of 3.19g (10.00mmol) of Boc-Arg (NO2) and 150mL of anhydrous tetrahydrofuran was added 1.35g (10.00mmol) of HOBt under ice-cooling, and after stirring for 5min, 2.27g (11.00mmol) of DCC was added, followed by stirring for 30min under ice-cooling to obtain reaction solution A. 3.50g (12.00mmol) of HCl.Gly-OBzl was added to the reaction solution A under ice-cooling, and the pH was adjusted to 9 with N-methylmorpholine.The reaction mixture was stirred at rt for 20h and TLC (dichloromethane/methanol ═ 20:1) showed the disappearance of Boc-Arg (NO 2). Insoluble matter in the reaction solution was filtered off, the filtrate was concentrated under reduced pressure, the residue was dissolved in 150mL of ethyl acetate, insoluble colorless solid was filtered off, and the filtrate was separately washed with saturated NaHCO3Aqueous solution (30 mL. times.3), saturated aqueous NaCl solution (30 mL. times.3), 5% KHSO4Aqueous solution (30 mL. times.3), saturated aqueous NaCl solution (30 mL. times.3), saturated aqueous NaHCO solution3Washing with water solution (30mL × 3), washing with saturated NaCl water solution (30mL × 3), drying the obtained ethyl acetate phase with anhydrous sodium sulfate for 12h, filtering to remove sodium sulfate, concentrating the filtrate under reduced pressure to obtain yellow syrup, dissolving with 30mL dichloromethane, standing at 4 deg.C for 12 h; a white solid precipitated and was filtered under reduced pressure to give 4.32g (93%) of the title compound as a colorless solid. ESI-MS (M/e) 467[ M + H]+。
EXAMPLE 6 preparation of Boc-Arg (NO2) -Gly
A solution of 2M NaOH was added dropwise to a solution of 4.32g (9.25mmol) of Boc-Arg (NO2) -Gly-OBzl in 30mL of methanol under ice-bath to adjust the pH of the solution to 13-14. The reaction mixture was stirred in an ice bath for 1h and TLC (dichloromethane/methanol/acetic acid, 20/1/2 drops, v/v/v) showed the disappearance of Boc-Arg (NO2) -Gly-OBzl. Dripping saturated KHSO in ice bath4Adjusting the pH of the solution to 7, and removing methanol by rotary removal under reduced pressure. The residue was added dropwise to saturated KHSO in an ice bath4The solution was adjusted to pH 2-3, extracted with ethyl acetate (50 mL. times.3), the ethyl acetate layers were combined, washed with saturated aqueous NaCl (30 mL. times.3), the resulting ethyl acetate phase was dried over anhydrous sodium sulfate for 12h, the sodium sulfate was filtered off, the filtrate was concentrated under reduced pressure, and 3.21g (92%) of the title compound was a colorless syrup.
EXAMPLE 7 preparation of Boc-Asp (OBzl) -Ser-OBzl
From 3.23g (10.00mmol) Boc-Asp (OBzl) and 2.78g (12.00mmol) HCl.Ser-OBzl, 4.43g (89%) of the title compound are obtained as yellow syrup according to the method of example 5. ESI-MS (M/e):501[ M + H]+。
EXAMPLE 8 preparation of HCl. Asp (OBzl) -Ser-OBzl
4.43g (8.86mmol) of Boc-Asp (OBzl) -Ser-OBzl was dissolved in 20mL of anhydrous ethyl acetate, and then 50mL of a solution of hydrogen chloride in ethyl acetate (4M) was added to the solution under ice-bath stirring at room temperature for 6h, and TLC (dichloromethane/methanol/acetic acid, 20/1/2 drops, v/v/v) showed disappearance of Boc-Asp (OBzl) -Ser-OBzl. The reaction solution was concentrated under reduced pressure, and the residue was redissolved with anhydrous ethyl acetate, then concentrated under reduced pressure, and redissolved with anhydrous ethyl acetate. This operation was repeated at least 3 times. The final residue was washed with dry ether to give 3.58g (93%) of the title compound as a yellow solid.
EXAMPLE 9 preparation of Boc-Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl
From 2.50g (6.65mmol) Boc-Arg (NO2) -Gly and 3.48g (7.97mmol) HCl. Asp (OBzl) -Ser-OBzl 2.55g (42%) of the title compound were obtained as colorless powder according to the method of example 5. ESI-MS (M/e):759[ M + H]+。
EXAMPLE 10 preparation of HCl.Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl
From 843mg (1.11mmol) Boc-Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl 729mg (94%) of the title compound was obtained as a colorless solid according to the method of example 8.
Example 11(2 'S, 5' S) -Tetrahydropyrazine [1 ', 2': 1,6] bis { (1R) - [ 1-Ethyl-Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl ] -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indoline } -1 ', 4' -dione (5)
168mg (0.35mmol) of (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6]And bis { (1R) - [ 1-Carbonylmethyl group]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione (4) was dissolved in 15mL of dichloromethane and the pH was adjusted to 9 with triethylamine under ice bath. 729mg (1.05mmol) of HCl Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl are dissolved in 15mL of dichloromethane, the pH is adjusted to 9 with triethylamine in ice bath, the solution is added dropwise to the dichloromethane solution of compound 4 in ice bath, 100mg of MgSO4. After the reaction mixture was stirred in ice bath for 1h, 27mg NaCNBH was added every 1h3Adding 108mg NaCNBH in total3. After that, TLC (dichloromethane/methanol ═ 7:1) was stirred at room temperature for 24h to show disappearance of compound 4, and the reaction was terminated. The reaction solution is respectively saturated NaHCO3Washing with aqueous solution (7 mL. times.3) and saturated aqueous NaCl solution (7 mL. times.3), drying the obtained dichloromethane phase with anhydrous sodium sulfate for 12h, filtering off sodium sulfate, concentrating the filtrate under reduced pressure, and purifying the obtained yellow solid by silica gel column chromatography (dichloromethane/methanol)20:1) to yield 52mg (8%) of the title compound as a light 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)。
EXAMPLE 12 preparation of (2 'S, 5' S) -Tetrahydropyrazine [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ] indoline } -1 ', 4' -dione (6)
50mg (0.03mmol) of (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1, 6%]And bis { (1R) - [ 1-ethyl-Arg (NO2) -Gly-Asp (OBzl) -Ser-OBzl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione (5) was dissolved in 1.5mL of trifluoroacetic acid. After that, 0.5mL of trifluoromethanesulfonic acid was added, and stirred for 30min under ice bath, and TLC (dichloromethane/methanol ═ 7:1) showed disappearance of compound 5. To the reaction mixture was added 25mL of anhydrous ether under ice-cooling, and the mixture was stirred for 10min, allowed to stand, and the supernatant was decanted. The residue was added with 25mL of anhydrous ether, stirred for 10min, allowed to stand, and the supernatant was decanted. This operation was repeated at least 3 times. The resulting yellow-black solid was dissolved in 1mL of 5% acetic acid and the solution was purified by Sephadex to give 6mg (16%) of the title compound as a pale 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)。
EXAMPLE 13 evaluation of anti-venous thrombosis Effect of (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione (6)
Experimental materials include warfarin sodium (CAS: 129-06-6, Bailingwei science and technology Co., Ltd.), urethane (CAS:51-79-6, Cat No. 30191228, national drug group chemical reagents Co., Ltd.), physiological saline (Shijiazhuang Siyao Co., Ltd.), trisodium citrate dihydrate (CMCNa, Cat No. 20170713, Beijing chemical plant).
Experimental animals SD strain rat, male, 250 + -20 g, purchased from Beijing Wittiulihua laboratory animals technology GmbH.
Experimental method the rat inferior vena cava ligation model is adopted in the experiment.
The grouping and dosage of (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione (Compound 6 for short) were 0.1. mu. mol/kg, the dosage of the positive control warfarin sodium was 4.87. mu. mol/kg, and the negative control was 0.5% CMCNa.
The preparation of the agent comprises the anesthetic which is a physiological saline solution (20%) of urethane, the compound 6 which is a suspension of 0.5% CMCNa, and the warfarin sodium which is a suspension of 0.5% CMCNa.
Experimental procedures rats were acclimatized and fasted for one day prior to surgery and were gavaged with a dose of 0.3mL/100g body weight. The administration is carried out 30min later and 2min before operation, and 20% urethane solution is used for abdominal cavity administration anesthesia. Fixing the rat on a rat fixing plate, preparing skin on the abdomen, sterilizing, opening the abdominal cavity along the leucorrhea line, and opening to expose one corner of the liver until the opening is about 4cm long. The organs such as small intestine in the abdominal cavity were removed and wrapped with gauze soaked with normal saline. Blunt separating connective tissue around blood vessel, exposing inferior vena cava and its branch, peeling off abdominal aorta and inferior vena cava below left renal vein, ligating inferior vena cava with suture soaked with physiological saline at junction of inferior vena cava and left renal vein, moving intestine and other organs back to abdominal cavity according to anatomical position, and suturing abdominal cavity layer by layer with suture.
After operation, the rat is placed in an environment with the temperature of 25-28 ℃ for circulation for 4 hours, the abdominal cavity is opened, the branches of the rat are tied one by one, the 2cm inferior vena cava is taken out from the tying position of the junction of the inferior vena cava and the left renal vein, and the thrombus is taken out from the inferior vena cava. The thrombus was weighed and the results were counted using the t-test. The operation was performed alternately with four of each group. The experimental data are shown in table 1.
the data in Table 1 show that compound 6 not only significantly inhibited venous thrombosis in rats at an oral dose of 0.1. mu. mol/kg, but also had activity comparable to 4.87. mu. mol/kg warfarin sodium. The present invention has an unexpected technical effect.
TABLE 1 anti-thrombotic Activity of Compound 6
Compound (I) Dosage form Thrombus weight (mean. + -. SD mg)
0.5％CMCNa 3mL/kg 21.74±4.51
Warfarin sodium 4.87μmol/kg 14.52±5.17
Compound 6 0.1μmol/kg 11.36±5.31a
a) P <0.01 to 0.5% CMCNa, and P >0.05 to 4.87. mu. mol/kg warfarin sodium; n is 8.

Claims (3)

1. Arg-Gly-Asp-Ser modified heptacyclic aldehyde of the formula, namely (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione,
2. A process for the preparation of (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione of claim 1 which comprises:
1) Carrying out Pictet-Spengler condensation on L-tryptophan benzyl ester and 1,1,3, 3-tetramethoxypropane under the catalysis of trifluoroacetic acid to obtain (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-benzyl carboxylate;
2) carrying out hydrogenolysis debenzylation on (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylic acid benzyl ester in methanol under the catalysis of Pd/C to obtain (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro-beta-carboline-3-carboxylic acid;
3) Intermolecular condensation of (3S) -1- (2, 2-dimethoxyethyl-2-yl) -2,3,4, 9-tetrahydro- β -carboline-3-carboxylic acid in anhydrous DMF (N, N-dimethylformamide) in the presence of benzotriazole-N, N ' -tetramethyluronium hexafluorophosphate to (2 ' S,5 ' S) -tetrahydropyrazino [1 ', 2 ': 1,6] bis { (1R) - [ 1-dimethoxyethyl-2-yl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indolino } -1 ', 4 ' -dione;
4) Using glacial acetic acid as a solvent, concentrated hydrochloric acid and water as catalysts to convert (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-dimethoxyethyl-2-yl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indoline } -1 ', 4' -dione into (2 'S, 5' S) -tetrahydropyrazino [1 ', 2': 1,6] bis { (1R) - [ 1-carbonylmethyl ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione;
5) Liquid phase condensation method for synthesizing HCl & Arg (NO) by using dicyclohexylcarbodiimide as condensing agent and 1-hydroxybenzotriazole as catalyst2)-Gly-Asp(OBzl)-Ser-OBzl；
6) Using sodium cyanoborohydride as a reducing agent to react HCl & Arg (NO)2) -Gly-Asp (OBzl) -Ser-OBzl to (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6]And bis { (1R) - [ 1-Carbonylmethyl group]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]Synthesis of (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6] on the aldehyde group of indole } -1 ', 4' -dione]and bis { (1R) - [ 1-ethyl-Arg (NO)2)-Gly-Asp(OBzl)-Ser-OBzl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione;
7) (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1,6] in a mixed solvent of trifluoroacetic acid and trifluoromethanesulfonic acid]and bis { (1R) - [ 1-ethyl-Arg (NO)2)-Gly-Asp(OBzl)-Ser-OBzl]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]and removing nitro and benzyl ester from indole } -1 ', 4' -diketone to obtain (2 'S, 5' S) -tetrahydropyrazine [1 ', 2': 1, 6)]And bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser]-2,3,4, 9-tetrahydro-1H-pyridine [3,4-b ]]And indole } -1 ', 4' -dione.
3. Use of (2 'S, 5' S) -tetrahydropyrazinyl [1 ', 2': 1,6] bis { (1R) - [ 1-ethyl-Arg-Gly-Asp-Ser ] -2,3,4, 9-tetrahydro-1H-pyrido [3,4-b ] indole } -1 ', 4' -dione according to claim 1 for the preparation of an anti-venous thrombosis medicament.
CN201810561696.3A 2018-06-04 2018-06-04 RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof Expired - Fee Related CN110563799B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810561696.3A CN110563799B (en) 2018-06-04 2018-06-04 RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810561696.3A CN110563799B (en) 2018-06-04 2018-06-04 RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof

Publications (2)

Publication Number Publication Date
CN110563799A true CN110563799A (en) 2019-12-13
CN110563799B CN110563799B (en) 2021-07-02

Family

ID=68771901

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810561696.3A Expired - Fee Related CN110563799B (en) 2018-06-04 2018-06-04 RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof

Country Status (1)

Country Link
CN (1) CN110563799B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113754724A (en) * 2020-06-04 2021-12-07 首都医科大学 Synthesis, bioactivity and application of dimethyl dioxane-tetrahydro-beta-carboline-3-formyl-RGDS

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103450334A (en) * 2012-05-29 2013-12-18 首都医科大学 RGD peptide-modified carbolino-hexahydropyrazine-1,4-diketones and their preparation method, antithrombotic effect and use
CN107488212A (en) * 2016-06-13 2017-12-19 首都医科大学 The O acetyl RGD tetrapeptides of warfarin 4, it is synthesized, activity and application
CN109912693A (en) * 2017-12-12 2019-06-21 首都医科大学 Seven ring aldehyde of RGDS modification, synthesis, anti-thrombus activity and application

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103450334A (en) * 2012-05-29 2013-12-18 首都医科大学 RGD peptide-modified carbolino-hexahydropyrazine-1,4-diketones and their preparation method, antithrombotic effect and use
CN107488212A (en) * 2016-06-13 2017-12-19 首都医科大学 The O acetyl RGD tetrapeptides of warfarin 4, it is synthesized, activity and application
CN109912693A (en) * 2017-12-12 2019-06-21 首都医科大学 Seven ring aldehyde of RGDS modification, synthesis, anti-thrombus activity and application

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
HAIMEI ZHU等: "Design, synthesis and evaluation of a novel π-π stacking nano-intercalator as an anti-tumor agent", 《MEDCHEMCOMM》 *
HAIMEI ZHU等: "Docking of THPDTPI：to explore P-selectin as a common target of anti-tumor, anti-thrombotic and anti-inflammatory agent", 《ONCOTARGET》 *
刘南等主编: "《血液系统疾病综合诊疗要点 上》", 30 April 2016, 吉林科学技术出版社 *
李家增，贺石林，王鸿利主编: "《临床血栓病学》", 31 October 2014, 上海交通大学出版社 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113754724A (en) * 2020-06-04 2021-12-07 首都医科大学 Synthesis, bioactivity and application of dimethyl dioxane-tetrahydro-beta-carboline-3-formyl-RGDS
CN113754724B (en) * 2020-06-04 2023-12-19 首都医科大学 Synthesis, biological activity and application of dimethyl dioxane-tetrahydro-beta-carboline-3-formyl-RGDS

Also Published As

Publication number Publication date
CN110563799B (en) 2021-07-02

Similar Documents

Publication Publication Date Title
CN109912597B (en) Heptacyclic aldehyde, its synthesis, antithrombotic activity and use
CN107686483B (en) Heptacyclic acetals, their preparation, antithrombotic activity and use
CN110577530B (en) Heptacyclic aldehyde, its synthesis, antithrombotic activity and use
CN100494195C (en) Carboline carboxylate derivative, and its synthesizing method and use
CN109912693B (en) RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
CN110563799B (en) RGDS modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
CN110577583B (en) RGDF modified heptacyclic aldehyde, its synthesis, antithrombotic activity and application
CN109912694B (en) RGDF modified heptacyclic aldehyde, its synthesis, antithrombotic activity and application
CN109912692B (en) YIGSK modified heptacyclic aldehyde, preparation, antithrombotic activity and application thereof
CN110551126B (en) Amino acid modified S, R-heptacyclic aldehyde, and synthesis, activity and application thereof
CN110577569B (en) RGDV-modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
CN110577571B (en) YIGSK modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
CN110577582B (en) LDV modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
CN108976285B (en) Gly-Pro-Arg-Pro-AA modified warfarin, synthesis, activity and application thereof
CN109912695B (en) RGDV-modified heptacyclic aldehyde, synthesis, antithrombotic activity and application thereof
WO2018068696A1 (en) Pegylated sinomenine and derivative thereof and preparation method and use of same
CN110551127B (en) Amino acid modified SS-heptacyclic aldehyde, preparation, activity and application thereof
CN110577570B (en) RGD tetrapeptide modified S, R-heptacyclic aldehyde, and synthesis, activity and application thereof
CN101265292B (en) Polypeptides substances, preparing method and use thereof
CN112010937B (en) YIGSR modified pentacyclic piperazinedione and preparation and application thereof
CN108948139B (en) warfarin-4-O-acetyl-Arg-AA compounds, their synthesis, activity and use
CN113754724B (en) Synthesis, biological activity and application of dimethyl dioxane-tetrahydro-beta-carboline-3-formyl-RGDS
CN112010930B (en) RGD modified pentacyclic piperazinedione and preparation and application thereof
CN108948140B (en) warfarin-4-O-acetyl-Arg-AA eleven compounds, and synthesis, activity and application thereof
CN111995658B (en) LDV (laser direct structuring) modified pentacyclic piperazinedione and preparation and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20210702

CF01 Termination of patent right due to non-payment of annual fee