CN107488210B - Warfarin-4-O-acetyl-dipeptide, its synthesis, activity and application - Google Patents
Warfarin-4-O-acetyl-dipeptide, its synthesis, activity and application Download PDFInfo
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- CN107488210B CN107488210B CN201610412840.8A CN201610412840A CN107488210B CN 107488210 B CN107488210 B CN 107488210B CN 201610412840 A CN201610412840 A CN 201610412840A CN 107488210 B CN107488210 B CN 107488210B
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- warfarin
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06086—Dipeptides with the first amino acid being basic
- C07K5/06095—Arg-amino acid
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06104—Dipeptides with the first amino acid being acidic
- C07K5/06113—Asp- or Asn-amino acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg, discloses a preparation method thereof, discloses anti-arterial thrombosis activity thereof, discloses activity thereof for reducing the content of vitamin K in vivo, discloses activity thereof for reducing the content of blood coagulation factor II in vivo, and discloses activity thereof for inhibiting platelet aggregation. Therefore, the invention discloses the application of the compounds in preparing anti-arterial thrombosis medicaments, the application in preparing anti-venous thrombosis medicaments, the application in preparing medicaments for inhibiting platelet aggregation, the application in preparing vitamin K antagonists and the application in preparing blood coagulation factor II antagonists.
Description
Technical Field
The present invention relates to warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg, to a process for their preparation, to their anti-arterial thrombotic activity, to their anti-thrombogenic activity, to their effect in reducing the level of vitamin K in the body, to their effect in reducing the level of coagulation factor II in the body, to their effect in inhibiting platelet aggregation and to their effect in down regulating the expression of the platelet membrane glycoprotein IIb/IIIa (GP IIb/IIIa) receptor in the body. Therefore, the invention relates to the application of the vitamin K antagonist in preparing the anti-arterial thrombosis medicine, the anti-venous thrombosis medicine, the platelet aggregation inhibiting medicine, the vitamin K antagonist and the blood coagulation factor II antagonist. The invention belongs to the field of biological medicine.
Background
Both arterial and venous thrombosis have become diseases with a high incidence of morbidity and mortality. Wherein the venous thrombosis mainly comprises 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. Because the incidence of arterial thrombosis and venous thrombosis increases exponentially with age, the threat of the two diseases to the health of people in the aging countries in China is particularly serious. If the population cardinality is considered, the absolute negative influence on the national civilization of China is particularly serious. Therefore, prevention and treatment of arterial thrombosis and venous thrombosis have been the focus of attention in the field of medicine. Although warfarin was used in clinical practice as a representative drug in 1941, its activity is manifested by a common complex role of its pharmacokinetic profile, coagulation factor metabolism, and availability of vitamin K. The dose-response relationship of warfarin shows great variability, which indicates that there is a great dose range for the anticoagulant therapy of warfarin and that there are great individual differences. Due to the narrow window of warfarin, under-dosing may lead to pulmonary embolism, while overdosing risks fatal bleeding. A great deal of structural modification is carried out on warfarin for more than 50 years in the past, but analogs with strong anti-thrombus activity and low bleeding side effect cannot be obtained. Contrary to the conventional thinking, the aim of the inventor for modifying the structure of warfarin is to convert warfarin into an analogue with double activities of resisting arterial thrombosis and resisting arterial thrombosis. However, it has not been satisfactory. After 5 years of exploration, the inventors found that the 4 th position of warfarin is modified by acetyl-Arg-Asp or acetyl-Arg-Asp, and the generated warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg show excellent anti-platelet aggregation activity on an in vitro anti-platelet aggregation model. Obviously, they are not prodrugs of warfarin. The antitarthrombosis activity of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg is 167 times stronger than that of aspirin, and the antitarthrombosis activity is 48.7 times stronger than that of warfarin. It can be seen that the present invention has significant technical advances. Thus, the inventors have proposed the present invention.
Disclosure of Invention
The first aspect of the present invention provides warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg.
The second aspect of the present invention provides a method for synthesizing warfarin derivatives, comprising:
1. synthesizing warfarin-4-O-benzyl acetate;
2. synthesizing warfarin-4-O-acetic acid;
3. uses protected amino acid as raw material, adopts DCC as condensing agent and HOBt as catalyst to synthesize Boc-Arg-Asp (OBzl) -OBzl and Boc-Asp (OBzl) -Arg (NO) by means of stepwise condensation2) -OBzl. Removing Boc protecting group with 4N hydrogen chloride in ethyl acetate to obtain Arg-Asp (OBzl) -OBzl and Asp (OBzl) -Arg (NO)2)-OBzl；
4. warfarin-4-O-acetic acid with Arg-Asp (OBzl) -OBzl or Asp (OBzl) -Arg (NO)2) OBzl coupling to warfarin-4-O-acetyl-Arg-Asp (OBzl) -OBzl or warfarin-4-O-acetyl-Asp (OBzl) -Arg (NO)2)-OBzl；
5. Removing the protecting group to obtain warfarin-4-O-acetyl-Arg-Asp or warfarin-4-O-acetyl-Asp-Arg;
the third aspect of the present invention is to evaluate the antithrombotic effects of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg.
The fourth aspect of the present invention was to evaluate the antithrombotic effects of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg.
The fifth aspect of the present invention was to evaluate the inhibition of platelet aggregation induced by Platelet Activating Factor (PAF), Thrombin (TH) and Adenosine Diphosphate (ADP) by warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg.
The sixth aspect of the present invention was to evaluate the effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on reducing vitamin K content in vivo.
The seventh aspect of the present invention is to evaluate the effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on reducing the level of coagulation factor II in vivo.
Drawings
FIG. 1 synthetic route to warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg (i) bromo-2-benzyl acetate, acetone, K2CO3,45℃；(ii)CH3OH,Pd/C,H2(ii) a (iii) DCC, HOBt, NMM, THF; (iv)4N hydrogen chloride in ethyl acetate.
FIG. 2 the anti-arterial thrombotic activity of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg, n ═ 10;
FIG. 3 warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg for anti-thrombogenic activity, n is 10;
FIG. 4 the effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on vitamin K content in vivo, n-5;
FIG. 5 Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on F II levels in rats, n-5.
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 warfarin-4-O-benzyl acetate
Placing 3.31g (10.00mmol) of warfarin in a 100mL eggplant bottle, adding about 40mL of acetone, not completely dissolving the acetone, heating in a 45 ℃ oil bath and stirring until the warfarin is dissolved, adding 1.73mL (11.00mmol) of benzyl bromide-2-acetate, continuing the reaction under the 45 ℃ oil bath, and after about 1h, finding a white solid attached to the bottle wall. After 48h reaction, the progress of the reaction was monitored by thin layer chromatography (TLC, petroleum ether/ethyl acetate 2:1), warfarin disappeared, the colorless solid produced by the reaction was filtered off, acetone was removed under reduced pressure to give a pale yellow oil,purification by silica gel column chromatography (petroleum ether/ethyl acetate 8:1) gave 3.02g (65%) of the title compound as a colourless solid. ESI-MS (M/e) 457[ M + H]+；1H-NMR(300MHz,DMSO-d6)δ/ppm＝7.89(dd,J1＝3.0Hz,J2＝9.0Hz,1H),7.63(dt,J1＝3.0Hz,J2＝9.0Hz,1H),7.43～7.31(m,9H),7.24(t,J＝9.0Hz,2H),7.15(tt,J＝9.0Hz,1H),5.26(s,2H),5.61(s,1H),5.02(d,J＝15.0Hz,1H),4.85(d,J＝15.0Hz,1H),4.97(t,J＝9.0Hz,1H),3.45(dq,J1＝9.0Hz,J2＝18.0Hz,2H),2.11(s,3H)。
EXAMPLE 2 preparation of warfarin-4-O-acetic acid
2.26g (4.95mmol) of warfarin-4-O-benzyl acetate was dissolved in 20mL of methanol, 220mg of palladium on carbon (Pd/C) was added, air was pumped out of a water pump with stirring, hydrogen was introduced, and the operation was repeated 3 times with stirring at room temperature with hydrogen. The reaction was monitored by TLC for completion, Pd/C was removed by filtration, the filtrate was concentrated under reduced pressure to remove the solvent, and the residue was solidified with petroleum ether and triturated with anhydrous ether to give 1.72g (93%) of the title compound as a colorless solid. ESI-MS (M/e):367[ M + H]+；1H-NMR(300MHz,DMSO-d6):δ/ppm＝12.86(s,1H),7.90(d,J＝6.0Hz,1H),7.63(t,J＝6.0Hz,1H),7.43～7.34(m,4H),7.27(t,J＝9.0Hz,2H),7.17(t,J＝9.0Hz,1H),4.99(t,J＝9.0Hz,1H),4.75(q,J1＝15.0Hz,J2＝30.0Hz,2H),3.54～3.47(m,2H),2.14(s,3H)。
EXAMPLE 3 preparation of Boc-Arg-Asp (OBzl) -OBzl
3.01g (10.98mmol) of Boc-Arg was added to a 250mL eggplant flask, which was dissolved in 150mL of anhydrous tetrahydrofuran, and 1.47g (10.89mmol) of HOBt and 2.70g (13.16mmol) of DCC were added thereto in an ice bath (0 ℃ C.) and activated for 30 min. A large amount of Dicyclohexylurea (DCU) precipitated. Dissolving 3.42g (10.93mmol) of Tos. Asp (OBzl) -OBzl in 100mL of anhydrous tetrahydrofuran, adding the mixture into the reaction solution under ice bath, adjusting the pH value to 8-9 by using N-methylmorpholine (NMM), stirring the mixture at room temperature for 6h, monitoring the completion of the reaction by TLC (dichloromethane/methanol 20:1), filtering DCU, removing the solvent from the filtrate under reduced pressure, dissolving the residue by using 100mL of ethyl acetate, filtering the insoluble DCU, and respectively using saturated Na for the filtrate2CO3Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated KHSO4Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated Na2CO3The solution (40 mL. times.3) was washed with a saturated NaCl solution (40 mL. times.3), and the resulting ethyl acetate phase was dried over anhydrous sodium sulfate for 2 hours. The sodium sulfate was filtered off, the solvent was removed from the filtrate under reduced pressure to give a colorless solid, which was purified by silica gel column chromatography (dichloromethane/methanol 50:1) to give 6.10g (97%) of a colorless solid product. ESI-MS (M/e):367[ M + H]+。
EXAMPLE 4 preparation of HCl Arg-Asp (OBzl) -OBzl
1.50g (2.63mmol) of Boc-Arg-Asp (OBzl) -OBzl was weighed into a 250mL eggplant flask, 10mL of anhydrous ethyl acetate was added thereto, and stirred to be completely dissolved, and 70mL of a 4N ethyl acetate solution of hydrogen chloride was added thereto under ice salt bath (-10 ℃ C.), and stirring was continued under ice bath. TLC (dichloromethane/methanol ═ 20:1) showed that the reaction was complete, the reaction solution was concentrated under reduced pressure using a water pump, the residue was redissolved with anhydrous ethyl acetate and concentrated under reduced pressure for 3 times to give a yellow oil, which was then soaked with anhydrous ether and concentrated under reduced pressure for 3 times to give 1.30g (97%) of a pale yellow syrup.
EXAMPLE 5 preparation of Boc-Asp (OBzl) -Arg (NO)2)-OBzl
3.04g (9.41mmol) of Boc-Asp (OBzl) was added to a 250mL eggplant flask, which was dissolved with 150mL of anhydrous tetrahydrofuran, and 1.26g (9.33mmol) of HOBt and 2.31g (11.21mmol) of DCC were added under ice bath (0 ℃ C.) to activate for 30 min. There was a significant amount of DCU evolved. 4.02g (9.27mmol) of HCl.Arg (NO)2) -OBzl is dissolved in 100mL of anhydrous tetrahydrofuran, added into the reaction solution in ice bath, the pH value is adjusted to 8-9 by NMM, stirred at room temperature for reaction for 6h, then TLC (dichloromethane/methanol ═ 20:1) is used for monitoring the reaction progress, the raw material point disappears, DCU is filtered out, the solvent is removed from the filtrate under reduced pressure, the residue is dissolved in 100mL of ethyl acetate, the insoluble DCU is filtered out, and the filtrate is respectively dissolved in saturated Na2CO3Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated KHSO4Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated Na2CO3The solution (40 mL. times.3) was washed with a saturated NaCl solution (40 mL. times.3), and the resulting ethyl acetate phase was usedDrying with anhydrous sodium sulfate for more than 2 h. The sodium sulfate was filtered off, the solvent was removed from the filtrate under reduced pressure to give a pale yellow solid, which was purified by silica gel column chromatography (dichloromethane/methanol 50:1) to give 5.10g (96%) of a colorless solid product. ESI-MS (M/e) 614[ M + H]+。
EXAMPLE 6 preparation of HCl. Asp (OBzl) -Arg (NO)2)-OBzl
Weighing 1.52g (2.48mmol) Boc-Asp (OBzl) -Arg (NO)2) OBzl in a 250mL eggplant flask, 10mL of anhydrous ethyl acetate was added to dissolve completely, 50mL of 4N ethyl acetate solution of hydrogen chloride was added under ice salt bath (-10 ℃ C.), and the reaction was stirred continuously under ice bath. TLC (dichloromethane/methanol ═ 20:1) showed that the reaction was complete, the reaction solution was concentrated under reduced pressure using a water pump, the residue was redissolved with anhydrous ethyl acetate and concentrated under reduced pressure for 3 times to give a yellow oil, which was then soaked with anhydrous ether and concentrated under reduced pressure for 3 times to give 1.30g (97%) of a pale yellow oil.
EXAMPLE 7 preparation of warfarin-4-O-acetyl-Arg-Asp (OBzl) -OBzl
1.05g (2.86mmol) of warfarin-4-O-acetic acid was added to a 100mL eggplant flask, and dissolved in 30mL of anhydrous tetrahydrofuran, and then 0.37g (2.74mmol) of HOBt and 0.68g (3.30mmol) of DCC were added thereto in an ice bath (0 ℃ C.) and activated for 30 min. There was a significant amount of DCU evolved. Dissolving 1.33g (2.63mmol) of HCl, Arg-Asp (OBzl) -OBzl in 30mL of anhydrous tetrahydrofuran, adding the mixture into the reaction solution in ice bath, adjusting the pH value to 8-9 by NMM, stirring the mixture at room temperature for 4 hours, monitoring the reaction progress by TLC (dichloromethane/methanol 20:1), filtering DCU, removing the solvent from the filtrate under reduced pressure, dissolving the residue with dichloromethane, filtering the insoluble DCU, and respectively using saturated Na for the filtrate2CO3Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated KHSO4Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated Na2CO3The solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3) was washed, and the methylene chloride phase was dried over anhydrous sodium sulfate for 2 hours. The sodium sulfate was filtered off, and the solvent was removed from the filtrate under reduced pressure to give a pale yellow oil, which was purified by silica gel column chromatography (dichloromethane/methanol ═ 30:1) to give 1.77g (79%) of a colorless solid. ESI-MS (M/e):818[ M + H]+；1H-NMR(300MHz,DMSO-d6):δ/ppm＝8.83(t,J＝7.2Hz,1H),8.50(dd,J1＝8.1Hz,J2＝13.5Hz,1H),7.90(m,1H),7.77(t,J＝5.4Hz,1H),7.63(t,J＝7.5Hz,1H),7.49～7.27(m,16H),7.25(m,2H),7.16(m,2H),5.10(s,2H),5.08(s,2H),4.90(t,J＝7.2Hz,1H),4.79(m,J＝6.6Hz,1H),4.63(s,2H),4.48(m,1H),3.44(m,2H),3.08(m,2H),2.86(dd,J1＝6.0Hz,J2＝16.8Hz,1H),2.96(dd,J1＝7.2Hz,J2＝16.8Hz,1H),2.13(s,3H),1.73～1.51(m,4H)。
EXAMPLE 8 preparation of warfarin-4-O-acetyl-Asp (OBzl) -Arg (NO)2)-OBzl
1.08g (2.95mmol) of warfarin-4-O-acetic acid was added to a 100mL eggplant flask, and dissolved in 30mL of anhydrous tetrahydrofuran, and then 0.37g (2.74mmol) of HOBt and 0.68g (3.30mmol) of DCC were added thereto in an ice bath (0 ℃ C.) and activated for 30 min. There was a significant amount of DCU evolved. 1.35g (2.67mmol) of HCl. Asp (OBzl) -Arg (NO)2) -OBzl is dissolved in 30mL of anhydrous tetrahydrofuran, added into the reaction solution in ice bath, the pH value is adjusted to 8-9 by NMM, stirred at room temperature for reaction for 4h, then TLC (dichloromethane/methanol ═ 20:1) is used for monitoring the reaction progress, the raw material point disappears, DCU is filtered out, the solvent is removed from the filtrate under reduced pressure, the residue is dissolved by ethyl acetate, the insoluble DCU is filtered out, and the filtrate is respectively saturated Na2CO3Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated KHSO4Solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3), saturated Na2CO3The solution (40 mL. times.3), saturated NaCl solution (40 mL. times.3) was washed, and the ethyl acetate phase was dried over anhydrous sodium sulfate for 2 hours. The sodium sulfate was filtered off, and the filtrate was concentrated under reduced pressure to give a pale yellow oil, which was purified by silica gel column chromatography (dichloromethane/methanol 50:1) to give 1.67g (75%) of a colorless solid. ESI-MS (M/e):863[ M + H]+；1H-NMR(300MHz,DMSO-d6):δ/ppm＝8.69(m,1H),8.57(d,J＝6.9Hz,1H),8.48(m,1H),7.81(dd,J1＝8.4Hz,J2＝12.3Hz,2H),7.61(t,J＝7.8Hz,1H),7.31(m,17H),7.25(m,1H),5.09(s,2H),5.06(m,2H),4.85(m,2H),4.52(m,2H),4.31(m,1H),3.40(m,2H),3.12(m,2H),2.78(m,2H),2.11(s,3H),1.78～1.53(m,4H).
EXAMPLE 9 preparation of warfarin-4-O-acetyl-Arg-Asp
1.12g (1.37mmol) of warfarin-4-O-acetyl-Arg-Asp (OBzl) -OBzl was dissolved in 50mL of methanol, 110mg of Pd/C was added, air was pumped out of a water pump with stirring, and hydrogen was introduced, and this operation was repeated 3 times, and the reaction was stirred with hydrogen at room temperature for about 10 hours. TLC (dichloromethane/methanol ═ 20:1) monitored the progress of the reaction, the starting material point disappeared, Pd/C was filtered off, the filtrate was freed of solvent under reduced pressure, it was solidified with petroleum ether to a white solid, which was purified by repeated trituration with anhydrous ether to yield 421mg (54%) of a white solid product. Mp:154-156 deg.C;
(c＝0.11,CH3OH)；ESI-MS(m/e):638[M+H]+；IR(cm-1):3334,1657,1608,1565,1453,1353,1101,1002,755,698；1H-NMR(300MHz,DMSO-d6):δ/ppm＝8.46(dd,J1＝7.8Hz,J2＝18.3Hz,1H),8.10(t,J＝6.6Hz,1H),7.90(dd,J1＝4.8Hz,J2＝7.8Hz,1H),7.69(m,1H),7.62(t,J＝8.4Hz,2H),7.44～7.34(m,5H),7.26(t,J＝7.2Hz,3H),7.17(m,2H),4.90(m,1H),4.65(s,2H),4.45(q,J＝6.0Hz,1H),4.22(m,1H),3.46(m,2H),3.11(m,2H),2.57(m,1H),2.35(dd,J1＝2.4Hz,J2＝15.6Hz,1H),2.14(s,3H),1.80～1.54(m,4H).
EXAMPLE 10 preparation of warfarin-4-O-acetyl-Asp-Arg
1.03g (1.19mmol) of warfarin-4-O-acetyl-Asp (OBzl) -Arg (NO2) -OBzl was dissolved in 50mL of methanol, 100mg of Pd/C was added, air was pumped through a water pump with stirring, and hydrogen was introduced, and this operation was repeated 3 times, and the reaction was stirred at room temperature with hydrogen for about 27 hours. TLC (dichloromethane/methanol ═ 20:1) monitored the progress of the reaction, the starting material point disappeared, Pd/C was filtered off, the filtrate was freed of solvent under reduced pressure, it was solidified with petroleum ether to a colorless solid, which was purified by C18 column chromatography to give 351mg (35%) of the product as a colorless solid. Mp 151-153 deg.C;
(c＝0.10,CH3OH)；ESI-MS(m/e):638[M+H]+；IR(cm-1):3328,3196,3069,2941,1656,1609,1565,1527,1493,1453,1397,1354,1274,1226,1197,1166,1102,1069,1017,911,893,756,699；1H-NMR(300MHz,DMSO-d6):δ/ppm＝8.80(m,1H),8.62(m,1H),7.89(m,1H),7.84(t,J＝7.5Hz,1H),7.63(t,J＝7.2Hz,2H),7.43～7.33(m,5H),7.25(t,J＝7.2Hz,3H),7.18～7.16(m,2H),4.88(m,1H),4.65(m,1H),4.12(m,1H),3.35～3.47(m,2H),3.01(m,2H),2.73～2.55(m,2H),2.14(s,3H),1.63～1.23(m,4H)
EXAMPLE 11 evaluation of the anti-arteriothrombotic Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg
Experimental Material
Urethane (CAS: 51-79-6, national drug group chemical Co., Ltd.), heparin sodium (CAS: 9041-08-1, Bailingwei science Co., Ltd.), and physiological saline (Shijiazhuang four drugs Co., Ltd.).
Laboratory animal
SD strain rats, male, 200 + -20 g, purchased from Experimental animals technologies, Inc. of Wei Tongli, Beijing.
Experimental method an arteriovenous bypass thrombosis model is adopted in the experiment.
Bypass cannula preparation
The bypass intubation consists of three sections, wherein a polyethylene tube with the inner diameter of 1.0mm and the outer diameter of 2.0mm is heated and drawn into a thin tube with one end being an oblique opening, the length of the thin tube is 10.0cm, the thin tube is respectively a right carotid vein intubation and a left carotid artery intubation and is positioned at two ends of the bypass intubation; the middle section is composed of a polyethylene pipe with the inner diameter of 3.5mm, and the fixed length is 8.0 cm; the length of the silk thread with rough surface is fixed to 6.0cm, and the silk thread with 4.0 plus or minus 0.1mg weight and the same rough degree is selected.
Silanization is carried out on the inner walls of the three sections of polyethylene tubes by using 1% of silicon ether solution (1% of silicon oil in ether solution), after the polyethylene tubes are completely dried, silk threads are placed in the middle section of polyethylene tubes in the carotid artery intubation direction, the three sections of polyethylene tubes are assembled and fixed by using a sealing film, and the tubes are filled with heparin before intubation.
Grouping and dosing
The dosages of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg of the compounds are 0.1 mu mol/kg; the positive control aspirin dose is 167 mu mol/kg and 16.7 mu mol/kg, and the negative control is physiological saline
Preparation of the used reagent
The anesthetic is 20% urethane aqueous solution prepared by normal saline, and the anticoagulant is 42mg/100mL heparin sodium aqueous solution prepared by normal saline.
Experimental procedures
Rats were each gavaged at a dose of 0.3mL/100g body weight, and anesthetized 30min later by abdominal injection of a 20% urethane solution (0.7mL/100 g). Fixing a rat on a rat fixing plate in a supine position, cutting the skin of the neck, separating a right common carotid artery and a left external jugular vein, ligating the distal ends of the right common carotid artery and the left external jugular vein respectively by using an operation line, cutting a V-shaped small opening on the exposed left external jugular vein, inserting the inclined port of the vein end of the bypass cannula manufactured on the upper side into the proximal end of the opening of the left external jugular vein, fixing a blood vessel and a polyethylene tube at the cannula position by using the operation line, accurately injecting heparin sodium water solution through the bypass cannula at the dose of 0.1mL/100g of body weight, and ensuring that an injector does not withdraw from the polyethylene tube. Clamping the proximal end of the right common carotid artery by an artery clamp, cutting a V-shaped small opening on the exposed artery, taking the tip of the polyethylene tube down from the injector, inserting the tube into the proximal end of the right common carotid artery, fixing the artery blood vessel and the polyethylene tube by an operation line, loosening the artery clamp and establishing an extracorporeal circulation bypass.
The body temperature of a rat and the blood flow in a bypass cannula are kept smooth, after the body circulation is carried out for 15min, the venous end cannula is firstly cut to observe whether the blood circulation is smooth, a thrombus thread is taken out from the arterial end of the cannula, and after floating blood on the thread is absorbed on filter paper, the wet weight of the filter paper is weighed and recorded, which represents the anti-arterial thrombosis activity. Data were counted using t-test.
Results of the experiment
The data are shown in FIG. 2. The results show that the thrombus weight of 0.1 mu mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rats (24.61 +/-4.56 mg, 23.95 +/-8.38 mg) is significantly less than the thrombus weight of saline treated rats (30.42 +/-3.49 mg, p <0.05), indicating that the compounds 0.1 mu mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg exhibit anti-arterial thrombus activity. Whereas aspirin at a dose of 16.7 μmol/kg did not have anti-arterial thrombotic activity, indicating that the compounds warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg have at least 167 times greater anti-arterial thrombotic activity than aspirin. This is an unexpected technical effect.
EXAMPLE 12 evaluation of the anti-venous-thrombotic Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg
Experimental Material
Uratan (ethyl carbamate, CAS: 51-79-6, national drug group chemical reagents Co., Ltd.), normal saline (Shijiazhuang four drugs Co., Ltd.) warfarin sodium (CAS: 129-06-6, Bailingwei science and technology Co., Ltd.);
laboratory animal
SD strain rats, male, 250 + -20 g, purchased from Experimental animals technologies, Inc. of Wei Tongli, Beijing.
Experimental methods the rat inferior vena cava ligation model was used for the experiments.
Grouping and dosing
The dosages of the compounds warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg of the invention are 0.1 mu mol/kg, the dosage of positive control warfarin is 4.87 mu mol/kg, and the negative control is normal saline.
Preparation of the used reagent
The anesthetic is a 20% urethane solution prepared from normal saline.
Experimental procedures
The experimental rats were acclimatized and fasted for one day before surgery, and were administered with gavage at a dose of 0.3mL/100g body weight, administered in a gavage manner, and administered 30min later with a 20% urethane solution 2min before surgery for intraperitoneal anesthesia. Rats were fixed on a rat fixing plate, and 2mL of blood was taken from the carotid artery and used for measurement of blood-related indices. Preparing skin of abdomen of rat, sterilizing, opening abdominal cavity along the leucorrhea line, descending to coagulated gland, and ascending to expose one corner of liver. The organs such as small intestine in abdominal cavity are pulled out to expose inferior vena cava, and the pulled-out organs are 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 renal vein, ligating inferior vena cava at junction of inferior vena cava and left renal vein with suture soaked with physiological saline, placing organs such as intestine back into abdominal cavity according to anatomical position, and suturing abdominal cavity layer by layer with suture.
After the operation, the rat is placed in an environment with the temperature of 25-28 ℃ for circulation for 4h, after 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 blood weight was calculated and the results were counted using the t-test. The operation was performed alternately with two per group. The experimental data are shown in FIG. 3.
Results of the experiment
The results show that the thrombus weight of 0.1 mu mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rats (12.72 +/-4.18 mg and 11.05 +/-2.61 mg) is significantly less than the thrombus weight of saline treated rats (22.93 +/-5.03 mg, p <0.01), indicating that 0.1 mu mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg exhibit anti-venous thrombosis activity. And the thrombus weights of the warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rats at the dose of 0.1. mu. mol/kg were not significantly different from the thrombus weights of the warfarin treated rats at the dose of 4.87. mu. mol/kg (12.12. + -. 3.86mg, p >0.05), indicating that the anti-thrombocyte activity of the compounds warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg was at least 48.7 times stronger than that of warfarin. This is an unexpected technical effect.
EXAMPLE 13 evaluation of the inhibitory Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on PAF, TH and ADP-induced platelet aggregation
Experimental Material
Sodium citrate (CAS: 68-04-2, national drug group chemical Co., Ltd.), physiological saline (Shijiazhuang Siyao Co., Ltd.), thrombin (TH, CAS:506-32-1, SIGMA reagent Co., Ltd.), adenosine diphosphate (ADP, CAS:58-64-0,
SIGMA reagent, Inc.), platelet activating factor (PAF, CAS:74389-68-7, SIGMA reagent, Inc.).
Laboratory instruments platelet aggregation apparatus: MODEL 700, CHRONO-LOG
Experimental fresh blood of porcine carotid artery
Experiment grouping
warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg, and the negative control is physiological saline. Preparation of the used reagent
The anticoagulant is 3.8% sodium citrate solution prepared from normal saline, the platelet activator is 50U/mL TH solution prepared from normal saline, 1mM ADP solution prepared from normal saline, and 5 × 10 solution prepared from normal saline-5mM PAF solution. The final concentration of inducer was 1U/mL TH, 20. mu. ADP M, PAF 1X 10-6mM。
Experimental methods
A blood collection bottle silanized and air dried one day in advance was charged with 50mL of an aqueous solution of sodium citrate (1: 9 ratio to blood). Collecting fresh pig carotid artery blood, shaking gently, rapidly centrifuging at 1000rpm for 10min, collecting supernatant to obtain Platelet Rich Plasma (PRP), centrifuging the rest blood at 3500rpm for 10min, and collecting supernatant to obtain Platelet Poor Plasma (PPP).
Add 500. mu.L of PPP to the cuvette on the platelet aggregation apparatus, to the PPP wells, add 480. mu.L of PRP to the cuvette, and to the PRP wells. The PRP used was incubated at 37 ℃ for 10min before measurement. After the instrument started running, the rotor was added and zeroed, 10 μ L of saline or compound aqueous solution was added to the PRP, followed by 10 μ L of platelet activation inducer. The change of the light transmittance is recorded, and the inhibition rate of the compound on platelet aggregation is calculated. The results are shown in Table 1.
Results of the experiment
TABLE 1 inhibition of platelet aggregation inhibition induced by PAF, TH, ADP (X + -SD%) by warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg
n is 3; final concentration of compound: 2X 10-5mol/L, the final concentration of inducer is: TH: 1U/mL, ADP: 20. mu. mol/L, PAF: 1X 10-6mmol/L。
Experimental results show that warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg can inhibit platelet aggregation induced by PAF, TH and ADP. Inhibition rates for PAF-induced platelet aggregation were 47.7% and 13.1%; the inhibition rate of TH-induced platelet aggregation is 23.5% and 8.1%; the inhibition rates for ADP-induced platelet aggregation were 40.4% and 45.8%. warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg are not prodrugs because they inhibit platelet aggregation in vitro. This is an unexpected technical effect.
EXAMPLE 14 evaluation of the Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on reducing vitamin K levels in rats
Experimental Material
Sodium citrate (CAS: 68-04-2, national drug group chemical Co., Ltd.), NS (Shijiazhuang Siyao Co., Ltd.), and distilled water.
Experimental sample
EXAMPLE 12 post-warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg rat carotid blood was given.
Experimental methods
Detection was performed using rat vitamin K1 elisa kit.
Sample collection
Collecting 0.1 mu mol/kg of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rat carotid artery blood by using a 3.8 percent sodium citrate solution as an anticoagulant, centrifuging the rat carotid artery blood for 15min at 1000rpm at 4 ℃ within 30min, and taking the supernatant as a sample for detection.
Standard preparation
And (3) taking the standard substance out of the kit, diluting 150 mu L of original-time standard substance by using 150 mu L of standard substance diluent, fully mixing to obtain a standard substance S5, arranging 4 centrifuge tubes of 1.5mL in sequence, adding 150 mu L of sample diluent respectively, sucking 150 mu L of standard substance S5 into an S4 centrifuge tube, blowing and uniformly mixing, and sequentially preparing an S5-S1 standard substance. The concentrations of the standard were S5(6ng/mL), S4(3ng/mL), S3(1.5ng/mL), S2(0.75ng/mL), S1(0.375ng/mL) and S0(0ng/mL), respectively.
A blank hole, a standard hole and a serum hole to be detected are respectively arranged. Adding 50 mu L of standard substance into the enzyme-labeled coated plate, adding 40 mu L of sample diluent into the serum hole to be detected, adding 10 mu L of serum to be detected, and slightly shaking and uniformly mixing. The plates were sealed with a sealing plate and incubated at 37 ℃ for 30 min. Carefully remove the coversheet membrane, discard the liquid, and wash the plate 5 times. Adding 50 mu L of enzyme labeling reagent into each hole except the blank hole, incubating for 30min at 37 ℃, washing, adding 50 mu L of color development agent A into each hole, adding 50 mu L of color development agent B, shaking gently, mixing uniformly, and developing for 10min at 37 ℃ in a dark place. The reaction was stopped by adding 50. mu.L of stop solution to each well, and the absorbance (OD value) of each well was measured at a wavelength of 450nm while the blank wells were zeroed within 15 min. A standard curve was drawn according to the OD value of the standard, and the concentration of the serum sample was calculated, and the data are shown in FIG. 4.
Results of the experiment
As can be seen from the data in FIG. 4, the vitamin K content of 0.1. mu. mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rats was significantly lower than that of saline treated rats (p <0.05), i.e., warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg reduced the vitamin K content in rats at a dose of 0.1. mu. mol/kg and was comparable to that of 4.87. mu. mol/kg warfarin treated rats. As can be seen, the activity of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg for reducing the vitamin K content in rats is 48.7 times stronger than that of warfarin. This is an unexpected technical effect.
EXAMPLE 17 evaluation of the Effect of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg on reducing the level of coagulation factor II in rats
Experimental Material
Sodium citrate (CAS: 68-04-2, national drug group chemical reagents, Inc.), physiological saline (Shijiazhuang Siyao, Inc.), pipette (Germany Prandde), distilled water;
experimental sample
EXAMPLE 12 post-warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg carotid arterial blood in rats
Experimental methods
The experiment was performed using a rat F II ELISA kit.
Collection of samples
Using 3.8% sodium citrate solution as anticoagulant, collecting 0.1 mu mol/kg of carotid artery blood of rats treated by warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg, centrifuging for 15min at 1000rpm at 4 ℃ within 30min, and taking supernatant (blood plasma) as a sample for detection.
Preparation of standard substance
And (3) taking the standard substance out of the kit, diluting 150 mu L of original-time standard substance by using 150 mu L of standard substance diluent, fully mixing to obtain a standard substance S5, arranging 4 centrifuge tubes of 1.5mL in sequence, adding 150 mu L of sample diluent respectively, sucking 150 mu L of standard substance S5 into an S4 centrifuge tube, blowing and uniformly mixing, and sequentially preparing an S5-S1 standard substance. The concentrations of the standard were S5(6ng/mL), S4(3ng/mL), S3(1.5ng/mL), S2(0.75ng/mL), S1(0.375ng/mL) and S0(0ng/mL), respectively.
A blank hole, a standard hole and a serum hole to be detected are respectively arranged. Adding 50 mu L of standard substance into the enzyme-labeled coated plate, adding 40 mu L of sample diluent into the serum hole to be detected, adding 10 mu L of serum to be detected, and slightly shaking and uniformly mixing. The plates were sealed with a sealing plate and incubated at 37 ℃ for 30 min. Carefully remove the coversheet membrane, discard the liquid, and wash the plate 5 times. Adding 50 mu L of enzyme labeling reagent into each hole except the blank hole, incubating for 30min at 37 ℃, washing, adding 50 mu L of color development agent A into each hole, adding 50 mu L of color development agent B, shaking gently, mixing uniformly, and developing for 10min at 37 ℃ in a dark place. The reaction was stopped by adding 50. mu.L of stop solution to each well, and the absorbance (OD value) of each well was measured at a wavelength of 450nm while the blank wells were zeroed within 15 min. A standard curve was drawn according to the OD value of the standard, and the concentration of the serum sample was calculated, and the data are shown in FIG. 5.
Results of the experiment
As can be seen from the data in FIG. 5, the FII content of 0.1. mu. mol/kg warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg treated rats was significantly lower (p <0.05) than that of saline treated rats, i.e., warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg reduced the FII content in rats at a dose of 0.1. mu. mol/kg and was comparable to that of 4.87. mu. mol/kg warfarin treated rats. As can be seen, the activity of warfarin-4-O-acetyl-Arg-Asp and warfarin-4-O-acetyl-Asp-Arg to reduce FII content in rats is 48.7 times stronger than that of warfarin. This is an unexpected technical effect.
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| CN102066559A (en) * | 2008-06-13 | 2011-05-18 | 北莱茵法威廉明斯特大学 | Biotechnological production of cyanophycin dipeptides |
| CN105273050A (en) * | 2014-07-10 | 2016-01-27 | 首都医科大学 | Imidazopyridine-6-formyl-amino acid benzyl esters, and synthesis, activity and application thereof |
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| CN102066559A (en) * | 2008-06-13 | 2011-05-18 | 北莱茵法威廉明斯特大学 | Biotechnological production of cyanophycin dipeptides |
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Non-Patent Citations (1)
| Title |
|---|
| Improvement of warfarin biopharmaceutics by conjugation with poly(ethylene glycol);MarinaZacchigna等;《European Journal of Pharmaceutical Science》;20041231;第23卷(第4-5期);摘要，第379页右栏第3段-第383页右栏第2段，图2 * |
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