CN113929735B - Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin, its synthesis, activity and use - Google Patents
Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin, its synthesis, activity and use Download PDFInfo
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Abstract
The invention discloses Gly-Pro-Arg-Pro-NHCH of the following formula 2 CH 2 NH-warfarin, its preparation, its anti-venous thrombotic activity, its advantage of no bleeding side effects, and its activity of reducing factor II, tissue factor/factor VII and the content of soluble fibrin monomer complex are disclosed. The invention thus discloses the use of the same in the preparation of an anti-venous thrombosis drug without bleeding side effects, in the preparation of a factor II antagonist, in the preparation of a tissue factor/factor VII antagonist and in the preparation of a soluble fibrin monomer complex antagonist.
Description
Technical Field
The invention relates to a Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin, to a process for its preparation, to its antithrombotic activity, further to its advantage of no bleeding side effects, and to its activity in reducing factor II, in reducing tissue factor/factor VII and in reducing the content of soluble fibrin monomer complexes. The invention thus relates to its preparation of an anti-static agent without bleeding side effectsPulse thrombosis medicine, its preparation method, its application in preparing blood coagulation factor II antagonist, its preparation method and its application in preparing tissue factor/blood coagulation factor VII antagonist and its preparation method. The invention belongs to the field of biological medicine.
Background
Thrombotic diseases are serious diseases threatening human life health, and have the characteristics of high morbidity and high mortality. Thrombotic diseases can be classified into arterial thromboembolic diseases and venous thromboembolic diseases according to the formation sites of the thrombus, wherein venous thromboembolic diseases (VTE) are the most common thrombotic diseases, and main diseases include Deep Vein Thrombosis (DVT) and Pulmonary Embolism (PE), mortality caused by PE is high, and serious complications can be caused if DVT is left to prevent. The main clinical treatment scheme of VTE is to use anticoagulant drug to perform anticoagulant treatment, and vitamin K1 antagonist represented by warfarin is one of the most commonly used anticoagulant drugs, but warfarin has the defects of large individual dosage difference, serious bleeding side effect, poor patient compliance and the like. Aiming at the defects of warfarin, the inventor discloses a modified warfarin (CN 107488157 A,CN 107488211 A,CN 107488214 A,CN 107488212 A,CN 107488213 A,CN 107488210 A,CN 107474030A). However, they are inhibitors of vitamin K and have no effect on the tissue factor/factor VII and soluble fibrin monomer complex which play an important role in the thrombotic process. In order to overcome the deficiency of this type of modified warfarin, the inventors have developed studies and searches for different objectives. Gly-Pro-Arg-Pro-NHCH was found later 2 CH 2 NH-warfarin has no effect on vitamin K, but rather acts on anti-venous thrombotic agents of the tissue factor/factor VII and soluble fibrin monomer complex. Based on these findings, the inventors have proposed the present invention.
Disclosure of Invention
The first aspect of the present invention provides a polypeptide of the formula Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin.
The second aspect of the present invention is to provide Gly-Pro-Arg-Pro-NHCH 2 CH 2 A method of preparing NH-warfarin, the method comprising:
1) Synthesizing warfarin-4-O-benzyl acetate;
2) Synthesizing warfarin-4-O-acetic acid;
3) Synthesis of Boc-NHCH 2 CH 2 NH-warfarin;
4) Boc-NHCH 2 CH 2 Conversion of NH-warfarin to HCl NH 2 CH 2 CH 2 NH-warfarin;
5) Synthesis of Boc-Gly-Pro-Arg (NO) 2 )-Pro-OBzl；
6) Boc-Gly-Pro-Arg (NO) 2 ) Conversion of Pro-OBzl to Boc-Gly-Pro-Arg (NO) 2 )-Pro；
7) HCl. NH 2 CH 2 CH 2 NH-warfarin and Boc-Gly-Pro-Arg (NO) 2 ) Synthesis of Boc-Gly-Pro-Arg (NO) by Pro reaction 2 )-Pro-NHCH 2 CH 2 NH-warfarin;
8) Boc-Gly-Pro-Arg (NO) 2 )-Pro-NHCH 2 CH 2 Conversion of NH-warfarin to Gly-Pro-Arg-Pro-NHCH as previously described 2 CH 2 NH-warfarin.
The third aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin anti-venous thrombosis effect.
The fourth aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin on bleeding time.
A fifth aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effects of NH-warfarin on clotting time.
A sixth aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin on international normalized ratio.
A seventh aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effects of NH-warfarin on plasma coagulation factor II content.
Eighth inner aspect of the present inventionThe appearance is that Gly-Pro-Arg-Pro-NHCH is evaluated 2 CH 2 Effects of NH-warfarin on plasma tissue factor/factor VII content.
A ninth aspect of the present invention is to evaluate Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effects of NH-warfarin on plasma soluble fibrin monomer complex content.
Drawings
FIG. 1 Gly-Pro-Arg-Pro-NHCH 2 CH 2 Synthetic route to NH-warfarin (i) benzyl bromo-2-acetate, acetone, K 2 CO 3 ,45℃；(ii)CH 3 OH,Pd/C,H 2 The method comprises the steps of carrying out a first treatment on the surface of the (iii) Dicyclohexylcarbodiimide, 1-hydroxybenzotriazole, N-methylmorpholine, tetrahydrofuran; (iv) an ethyl acetate solution of hydrogen chloride (4N); (v) acetone, sodium hydroxide solution (2N); (vi) trifluoroacetic acid, trifluoromethanesulfonic acid.
Detailed Description
To further illustrate the invention, a series of examples are given below. These examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention.
EXAMPLE 1 preparation of warfarin-4-O-benzyl acetate (1)
26.48g (80.00 mmol) of warfarin was dispersed in 400mL of acetone, stirred at 45℃until warfarin was dissolved, and 12.1g (88.0 mmol) of K was added to the resulting solution 2 CO 3 Then 14mL (88 mmol) of benzyl bromoacetate was added and the reaction was continued at 45℃for 96 hours, TLC (Petroleum ether/ethyl acetate, 2/1) showed the starting point to disappear, the filtrate was concentrated under reduced pressure, and the pale yellow oil was purified by silica gel column chromatography (petroleum ether/ethyl acetate, 8/1) to give 19.77g (54.2%) of the title compound as a colorless solid. ESI-MS (m/e): 457[ M+H ]] + ； 1 H NMR(300MHz,DMSO-d 6 )δ/ppm＝7.89(dd,J 1 ＝3.0Hz,J 2 ＝9.0Hz,1H),7.63(dt,J 1 ＝3.0Hz,J 2 ＝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,J 1 ＝9.0Hz,J 2 ＝18.0Hz,2H),2.11(s,3H)。
EXAMPLE 2 preparation of warfarin-4-O-acetic acid (2)
19.77g (43.36 mmol) of warfarin-4-O-benzyl acetate (1) are dissolved in 150mL of tetrahydrofuran, 4.94g of Pd/C are added, air is removed with stirring, hydrogen is introduced and stirring is carried out at room temperature for 72 hours. TLC (petroleum ether/ethyl acetate, 2/1) showed the starting material point disappeared, filtered and the filtrate concentrated under reduced pressure to give 15.58g (98.2%) of the title compound as a colorless solid. ESI-MS (m/e): 367[ M+H ]] + ； 1 H-NMR(300MHz,DMSO-d 6 ):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(dd,J 1 ＝15.0Hz,J 2 ＝30.0Hz,2H),3.54～3.47(m,2H),2.14(s,3H)。
EXAMPLE 3 preparation of Boc-NHCH 2 CH 2 NH-warfarin (3)
1.601g (4.374 mmol) of warfarin-4-O-acetic acid (2) was dissolved in 20mL of tetrahydrofuran, 0.5906g (4.375 mmol) of 1-hydroxybenzotriazole and 0.9831g (4.772 mmol) of dicyclohexylcarbodiimide were added at 0℃and activated with stirring under ice bath for 30min. Then 0.60mL (4.0 mmol) of Boc-ethylenediamine is sucked and added into the reaction solution under ice bath, the pH value of the reaction solution is regulated to 9 by using N-methylmorpholine, the reaction is stirred and carried out for 14 hours at room temperature, TLC (dichloromethane/methanol, 35/1 and 2d glacial acetic acid) shows that the raw material point disappears, the filtration and the concentration of the filtrate under reduced pressure are carried out, and the residues are respectively dissolved by using 100mL of ethyl acetate and then saturated NaHCO are respectively used 3 Washing with aqueous solution (40 mL. Times.3), washing with saturated aqueous NaCl solution (40 mL. Times.3), 5% KHSO 4 Washing with aqueous solution (40 mL. Times.3), washing with saturated aqueous NaCl solution (40 mL. Times.3), and saturated NaHCO 3 Aqueous washing (40 mL. Times.3), saturated aqueous NaCl washing (40 mL. Times.3), ethyl acetate phase and anhydrous Na 2 SO 4 Drying for 6 hours, filtration and concentration of the filtrate under reduced pressure gave crude pale yellow solid which was purified by column chromatography on silica gel (dichloromethane/methanol, 120/1) to give 1.57g (77%) of the title compound as colorless solid. 1 H NMR(300MHz,DMSO-d 6 ):δ/ppm＝8.39(s,1H),7.85(d,J＝7.8Hz,1H),7.65(t,J＝7.2Hz,1H),7.42(m,2H),7.36(d,J＝7.8Hz,2H),7.27(t,J＝7.2Hz,2H),7.19(m,1H),6.89(s,1H),4.90(t,J＝7.2Hz,1H),4.49(m,2H),3.46(d,J＝7.2Hz,2H),3.25(m,2H),3.10(m,2H),2.15(s,3H),1.38(s,9H)。
EXAMPLE 4 preparation of HCl NH 2 CH 2 CH 2 NH-warfarin (4)
1.452g (2.857 mmol) Boc-NHCH 2 CH 2 NH-warfarin (3) was dissolved in 15mL of dry ethyl acetate and 20mL of 4N strength ethyl acetate solution of hydrogen chloride was added under ice salt bath, followed by stirring for 4 hours. TLC (dichloromethane/methanol, 10/1 and 3 drops of glacial acetic acid) showed the disappearance of the starting material points, and the reaction solution was concentrated under reduced pressure to give a colorless viscous solid, which was uniformly dispersed with anhydrous diethyl ether, left to stand, and after decanting the upper layer solvent, concentrated under reduced pressure to give 1.29g (100%) of the title compound as a colorless solid. ESI-MS (m/e): 409[ M+H ]] + 。
EXAMPLE 5 preparation of Boc-Gly-Pro-OBzl
7.730g (44.15 mmol) of Boc-Gly are dissolved in 100mL of dry tetrahydrofuran, 5.940g (44.00 mmol) of 1-hydroxybenzotriazole and 9.888g (48.00 mmol) of dicyclohexylcarbodiimide are added at 0℃and stirred for 30 minutes. Then 9.665g (40.00 mmol) of HCl-Pro-OBzl was added, the pH of the reaction solution was adjusted to 9 with N-methylmorpholine, stirred at room temperature for 17 hours, TLC (dichloromethane/methanol, 20/1) showed the starting point disappeared, filtered, the filtrate was concentrated under reduced pressure, the residue was dissolved with 150mL of ethyl acetate, insoluble colorless solids were filtered off, and the filtrates were each treated with saturated NaHCO 3 Washing with aqueous solution (50 mL. Times.3), washing with saturated aqueous NaCl solution (50 mL. Times.3), 5% KHSO 4 Solution (50 mL. Times.3), saturated aqueous NaCl solution (50 mL. Times.3), saturated NaHCO 3 Washing with aqueous solution (50 mL. Times.3), washing with saturated aqueous NaCl solution (50 mL. Times.3), and washing the ethyl acetate phase with anhydrous Na 2 SO 4 Drying for 12 hours, filtration and concentration of the filtrate under reduced pressure gave a pale yellow solid which was purified by silica gel column chromatography (petroleum ether/ethyl acetate, 3/1) to give 13.42g (93%) of the title compound as a colorless solid. ESI-MS (m/e): 363[ M+H ]] + 。
EXAMPLE 6 preparation of Boc-Gly-Pro
4.774g (13.18 mmol) of Boc-Gly-Pro-OBzl was dissolved in 50mL of methanol, 1.0g of Pd/C was added, and the mixture was stirred and air was removed, followed by introducing hydrogen gas and stirring at room temperature for 48 hours. TLC (dichloromethane/methanol, 20/1) showed the starting material point disappeared, filtered and the filtrate concentrated under reduced pressure to give 3.411g (95%) of the title compound as a colorless solid. ESI-MS (m/e): 273[ M+H ]] + ； 1 H-NMR(300MHz,DMSO-d 6 ):δ/ppm＝6.83(t,J＝6.0Hz,1H),4.22(dd,J 1 ＝3.0Hz,J 2 ＝9.0Hz,1H),3.80(dd,J 1 ＝5.7Hz,J 2 ＝17.1Hz,1H),3.67(dd,J 1 ＝5.7Hz,J 2 ＝16.8Hz,1H),3.48(m,1H),3.38(m,1H),2.10(m,1H),1.91(m,1H),1.83(m,1H),1.38(s,9H)。
EXAMPLE 7 preparation of Boc-Arg (NO 2 )-Pro-OBzl
From 3.509g (11.00 mmol) Boc-Arg (NO) by the method of example 5 2 ) And 2.516g (10.41 mmol) HCl Pro-OBzl gave 4.05g (80%) of the title compound as a colorless solid product. ESI-MS (m/e): 507[ M+H ]] + ； 1 H-NMR(300MHz,DMSO-d 6 ):δ/ppm＝7.33(m,5H),6.98(d,J＝7.8Hz,1H),5.10(s,2H),4.40(m,1H),4.37(s,1H),3.92(m,1H),3.62(m,1H),3.10(s,2H),2.21(m,1H),1.94(t,J＝6.3Hz,2H),1.88(m,1H),1.56～1.45(m,4H),1.37(s,9H)。
EXAMPLE 8 preparation of HCl Arg (NO) 2 )-Pro-OBzl
From 1.80g (3.56 mmol) of Boc-Arg (NO) by the method of example 4 2 ) Pro-OBzl gave 1.53g (97%) of the title compound as a colorless solid. ESI-MS (m/e): 407[ M+H ]] + 。
EXAMPLE 9 preparation of Boc-Gly-Pro-Arg (NO) 2 )-Pro-OBzl
From 1.755g (6.45 mmol) of Boc-Gly-Pro and 2.566g (5.80 mmol) of HCl-Arg (NO) by the method of example 5 2 ) Pro-OBzl gave 2.31g (60%) of the title compound as a colorless solid. ESI-MS (m/e): 662[ M+H ]] + ； 1 H-NMR(300MHz,DMSO-d 6 ):δ/ppm＝8.49(s,1H),8.19(d,J＝7.5Hz,1H),7.35(m,5H),6.79(s,1H),5.10(t,J＝12.9Hz,1H),4.45～4.42(m,2H),4.37(dd,J 1 ＝4.5Hz,J 2 ＝8.1Hz,1H),3.83～3.66(m,2H),3.6～3.37(m,4H),3.11(s,2H),2.21(m,1H),1.94(t,J＝6.3Hz,2H),1.88(m,1H),1.56～1.45(m,4H),1.37(s,9H)。
EXAMPLE 10 preparation of Boc-Gly-Pro-Arg (NO) 2 )-Pro
2.656g (4.02 mmol) Boc-Gly-Pro-Arg (NO) 2 ) Pro-OBzl is dissolved in 35mL of acetone, and 2N sodium hydroxide water solution is added dropwise under ice bathThe reaction solution was adjusted to pH 13.0 and stirred for 6 hours, and TLC (dichloromethane/methanol, 10/1 and 3 drops of glacial acetic acid) showed the disappearance of the starting material points. Saturated KHSO under ice bath 4 The pH of the reaction mixture was adjusted to 7 with an aqueous solution, concentrated under reduced pressure, and the residue was taken up in 3mL of H 2 O is dissolved by saturated KHSO 4 The aqueous solution was adjusted to pH 2 and then extracted with ethyl acetate (30 mL. Times.4), the ethyl acetate phase was taken up in anhydrous Na 2 SO 4 Drying for 10 hours, filtration and concentration of the filtrate under reduced pressure gave 2.115g (92%) of the title compound as a pale yellow oil. ESI-MS (m/e): 571[ M+H ]] + 。
EXAMPLE 11 preparation of Boc-Gly-Pro-Arg (NO) 2 )-Pro-NHCH 2 CH 2 NH-warfarin (5)
From 1.141g (2.000 mmol) of Boc-Gly-Pro-Arg (NO) by the method of example 5 2 ) Pro and 0.8081g (1.817 mmol) HCl NH 2 CH 2 CH 2 NH-warfarin (4) gives 1.19g (68%) of the title compound as a colourless solid. 1 H NMR(300MHz,DMSO-d 6 )δ/ppm＝8.50(s,1H),8.39(s,1H),8.12(m,1H),7.93(s,1H),7.85(d,J＝7.2Hz,1H),7.65(m,1H),7.43(d,J＝7.8Hz,2H),7.34(m,2H),7.27(m,2H),7.19(m,1H),6.74(s,1H),4.90(m,1H),4.50(s,3H),4.35(m,1H),4.21(m,1H),3.74(m,2H),3.57(m,4H),3.47(d,J＝6.6Hz,2H),3.27(s,3H),3.16(s,3H),2.16(s,3H),2.00～1.57(m,12H),1.38(s,9H)。
EXAMPLE 12 preparation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin (6)
Ice salt bath and stirring were followed by dropwise addition of 7.5mL trifluoroacetic acid to 250mg (0.26 mmol) Boc-Gly-Pro-Arg (NO) 2 )-Pro-NHCH 2 CH 2 In NH-warfarin (5), 2.5mL of trifluoromethanesulfonic acid was added dropwise, and TLC (ethyl acetate/glacial acetic acid/water, 3/1/1) showed the disappearance of the starting material point after 1 hour of reaction. Immediately, the reaction mixture was concentrated under reduced pressure in an ice bath, the residue was diluted with 30mL of diethyl ether having been previously cooled, and after standing for 10 minutes, the upper clear solution was poured out, the residue was further diluted with 30mL of diethyl ether having been previously cooled, and after standing for 10 minutes, the upper clear solution was poured out again. This operation was repeated 3 times. Finally, the residue was concentrated under reduced pressure, and the resulting brown oil was taken up in 3mL H 2 Dissolving O, regulating pH to 7 with 10% ammonia water in ice bath, and filtering to obtainIs initially purified by Sephadex-G10 gel filtration chromatography, and the eluate monitored by TLC (ethyl acetate/glacial acetic acid/water, 3/1/1) with UV coloration is freeze-dried to give 170mg of yellow powder. The powder was dissolved in 2mL of 5% aqueous methanol using RP-C 18 Column chromatography purification (methanol/water, 1/1) and freeze-drying of the collected eluate afforded 113mg (53%) of the title compound as a colorless powder. ESI-MS (m/e): 816[ M+H ]] + ；(c＝0.13,CH 3 OH)； 1 H NMR(300MHz,DMSO-d 6 ):δ/ppm＝8.45(m,1H),8.28～8.00(m,5H),7.85(d,J＝7.8Hz,2H),7.64(t,J＝7.5Hz,1H),7.43(d,J＝9.0Hz,2H),7.35(m,3H),7.26(t,J＝7.5Hz,3H),7.19(m,2H),4.90(t,J＝7.2Hz,1H),4.51(m,2H),4.42(m,2H),4.21(m,1H),3.78(s,2H),3.58(m,4H),3.47(d,J＝7.2Hz,2H),3.26～3.14(m,6H),2.15(s,3H),2.02～1.57(m,12H)。
EXAMPLE 13 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Anti-venous thrombosis of NH-warfarin (6)
Experimental materials
Ulatan (urethane, CAS:51-79-6, national pharmaceutical Congress Chemicals Co., ltd.), warfarin sodium (CAS: 129-06-6, carbofuran technologies Co., ltd.).
Experimental animal
Male SD rats (250+ -20 g) purchased from Peking Vitre Liwa laboratory animal technologies Co. The evaluation was performed to prepare a rat inferior vena cava ligation model.
Dosage for administration
Gly-Pro-Arg-Pro-NHCH of the invention 2 CH 2 NH-warfarin (6) is dissolved in normal saline with the dosage of 0.82 mu mol/kg; the positive control warfarin is dissolved in normal saline with the dosage of 0.82 mu mol/kg; the negative control was normal saline.
Experimental operation
Rats were acclimatized and fasted for one day prior to surgery and were gavaged with compound 6 in saline at a dose of 0.82 μmol/kg; or physiological saline solution of warfarin with the dosage of 0.82 mu mol/kg; or physiological saline. Rats were anesthetized with 20% uratam solution intraperitoneally at 2 minutes prior to surgery 30 minutes after dosing. Then fixed on a rat fixing plate, the abdomen is prepared and disinfected, and then the abdominal cavity is opened along the abdominal white line until one corner of the liver is exposed, and the opening is about 4cm long. Organs such as small intestine in the abdominal cavity are removed and wrapped with gauze soaked with physiological saline. The connective tissue around the blood vessel is passively separated, the inferior vena cava and branches thereof are exposed, the abdominal aorta and the inferior vena cava are peeled off below the left renal vein, then the inferior vena cava is ligated at the junction of the inferior vena cava and the left renal vein by using a suture thread soaked by physiological saline, the organs such as intestines and the like are moved back to the abdominal cavity according to anatomical positions, the abdominal cavity is sutured layer by using the suture thread, and then the rat is placed in an environment of 25-28 ℃ for 4 days for circulation. Then receiving ether for anesthesia, opening the abdominal cavity, ligating each branch vessel of the inferior vena cava one by one, taking out the inferior vena cava 2cm from the ligature of the junction of the inferior vena cava and the left renal vein, and taking out the thrombus. Blood was left as a sample for the determination of factor II, tissue factor/factor VII and soluble fibrin monomer complex. Thrombus was weighed and the results were counted using T-test. The procedure was alternated with four per group. The thrombus was found in Table 1.
TABLE 1 treatment of venous thrombosis weight in rats
a) P is less than 0.01 compared with normal saline; n=8
The data in Table 1 show that compound 6 was effective in inhibiting venous thrombosis in rats at an oral dose of 0.82. Mu. Mol/kg for 4 days following treatment. It can be seen that the invention has remarkable technical effects.
EXAMPLE 14 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin (6) on bleeding time
The rats of example 13 were anesthetized prior to thrombus removal and then a 2mm deep wound was made with a scalpel at the rat's tail 8cm, while starting the timing, wiping blood with filter paper every 5 seconds, and stopping the timing when no blood trace was visible on the filter paper, the time noted being the rat tail bleeding time. Bleeding time was counted by means of T-test. The procedure was alternated with four per group. Bleeding time is shown in table 2.
TABLE 2 treatment of rat tail bleeding time
a) The ratio P of the water to the normal saline is less than 0.01; n=11
The data in Table 2 shows that compound 6 was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, and that the tail bleeding time of the rats was not different from that of the normal saline-treated rats, whereas warfarin was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, and that the tail bleeding time of the rats was significantly longer than that of the normal saline-treated rats, showing significant bleeding side effects. Compound 6 did not cause bleeding events similar to warfarin, and was safer than warfarin. It can be seen that the present invention has unexpected technical effects.
EXAMPLE 15 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin (6) on clotting time
The rat of example 13 was anesthetized before thrombus removal, its abdomen was laid down on a rat plate, the rat tail was rubbed with absorbent cotton ball dipped with medical alcohol to sterilize, 8cm was measured from the rat tail tip with a ruler and marked, a blood vessel located in the middle of the rat tail was found at a position approximately in the middle of the rat tail, a wound of about 5mm long and about 2mm deep was cut with a surgical blade, and immediately a drop of blood was taken with a glass plate and timing was started when blood was discharged. And repeatedly picking up blood drops by using the needle point of the syringe with the frequency of about 2 times/second until the needle point can pick up the filiform thrombus, and stopping timing, wherein the obtained time is the coagulation time of the rat. The obtained clotting time is counted by means of t-test. The procedure was alternated with four per group. The clotting times are shown in Table 3.
TABLE 3 clotting time for rats treated
a) P is less than 0.01 compared with normal saline; n=10
The data in Table 3 shows that compound 6 was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, the rat's rat tail clotting time was not different from that of normal saline-treated rats, whereas warfarin was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, the rat tail clotting time was significantly longer than that of normal saline-treated rats, showing significant bleeding side effects. Compound 6 did not cause bleeding events similar to warfarin, and was safer than warfarin. It can be seen that the present invention has unexpected technical effects.
EXAMPLE 16 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 The effect of NH-warfarin (6) on the international standardized ratio of rats the international standardized ratio reflects prothrombin time, which can be calculated automatically by a semiautomatic coagulometer (model TS6000, MD Pacific). The specific test method is that 3.6 mL/rat whole blood collected in example 13 is placed in a centrifuge tube filled with 0.4mL sodium citrate solution (3.8%), after centrifugation for 15 minutes at 2500g, upper-layer platelet plasma (PPP) is sucked, 100 mu L PPP is added in a test cup, a test magnetic bead is added, a sample to be tested is preheated for 180 seconds in a preheating zone of an instrument, 200 mu L prothrombin time test solution (kit goods number: 20-7011, MD Pacific) is added, the instrument starts to automatically detect prothrombin time, and an international standardized ratio is automatically calculated, and the statistical result is counted in a T test mode, wherein the international standardized ratio is shown in Table 4.
TABLE 4 International normalized ratio for rats treated
a) P is less than 0.01 compared with normal saline; n=10
The data in Table 4 show that compound 6 was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, the international normalized ratio of the rats was not different from that of the normal saline-treated rats, whereas warfarin was continuously treated for 4 days at an oral dose of 0.82. Mu. Mol/kg, and the international normalized ratio of the rats was significantly greater than that of the normal saline-treated rats, showing significant bleeding side effects. Compound 6 did not cause bleeding events similar to warfarin, and was safer than warfarin. It can be seen that the present invention has unexpected technical effects.
EXAMPLE 17 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin (6) on the plasma coagulation factor II (FIIa) content of rats
Whole blood of rats collected in example 13, 4.0 mL/mouse, was placed in a centrifuge tube containing 0.4mL of sodium citrate solution (3.8%), and after centrifugation at 1000g for 15 minutes, upper layer Platelet Poor Plasma (PPP) was aspirated. PPP samples of 6 rats after administration were taken for each experimental group, diluted 1:1 with the sample diluent in the kit, and added to 50ul in the reaction well. At the same time 50. Mu.L of diluted standard was added to the reaction well. 50. Mu.L of biotin-labeled antibody was immediately added. Covering the membrane plate, mixing with gentle shaking, and incubating at 37deg.C for 1 hr. And then the liquid in the holes is thrown off, each hole is filled with the washing liquid, the washing liquid is thrown off after shaking for 30 seconds, and the washing liquid is beaten by using absorbent paper. This operation was repeated 3 times. Then 80. Mu.L of streptavidin-HRP was added to each well, mixed gently with shaking, and incubated at 37℃for 30 minutes. And after the incubation is finished, the liquid in the holes is thrown off, the washing liquid is filled in each hole, the washing liquid is thrown off after shaking for 30 seconds, the washing liquid is beaten to dryness by using absorbent paper, and the operation is repeated for 3 times. Substrate A, B was added at 50. Mu.L each to each well, mixed gently with shaking, and incubated at 37℃for 10 minutes. Avoiding illumination, then taking out the ELISA plate, rapidly adding 50 mu L of stop solution, immediately measuring the result after adding the stop solution, and measuring the OD value of each hole at the wavelength of 450nm by using an ELISA. And subtracting the OD value read at 540nm from the OD value read at 450nm, drawing a standard curve by taking the OD value as an ordinate and the concentration of the blood coagulation factor II (FIIa) standard substance as an abscissa, and converting the FIIa content of the sample into the corresponding concentration from the standard curve according to the OD value. The differences in FIIa levels in the plasma of rats were analyzed by the one-way analysis of variance (LSD) method in SPSS22.0 and shown in Table 5.
TABLE 5 treatment of FIIa levels in rat plasma
a) P is less than 0.01 compared with normal saline; n=5
The data in Table 5 show that compound 6 was effective in reducing the levels of FIIa in rat plasma after 4 days of continuous oral administration at an oral dose of 0.82. Mu. Mol/kg. It can be seen that the invention has remarkable technical effects.
EXAMPLE 18 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin (6) on rat plasma TF/FVIIa levels
Whole blood of rats collected in example 13, 4.0 mL/mouse, was placed in a centrifuge tube containing 0.4mL of sodium citrate solution (3.8%), and after centrifugation at 1000g for 15 minutes, upper layer Platelet Poor Plasma (PPP) was aspirated. Each experimental group took 6 post-dose rats of PPP samples, each well was added with 100. Mu.L of each sample and standard, incubated at 37℃for 2 hours after covering with a cover plate membrane, all the liquid in the plates was poured, without washing the plates, 100. Mu.L of Biotin solution was directly added to each well, and incubated at 37℃for 1 hour after covering with a new cover plate membrane. The plate was then discarded, 200. Mu.L of wash solution was added to each well, and after 2min of standing, the wash solution was discarded and the 96-well plate was dried by pipetting, and this was repeated 3 times. Next, 100. Mu.L of HRP-avidin was added to each well and covered with a fresh cover film and incubated at 37℃for 1 hour. After the incubation, the plate washing operation was repeated 5 times to remove the liquid in each well. 90 mu L of 3,3', 5' -tetramethyl benzidine is added into each hole, the new cover plate film is covered, the mixture is incubated for 20 minutes at 37 ℃ in a dark place, then 50 mu L of stop solution is added into each hole, and the OD value of each hole is rapidly read at the dual wavelengths of 450nm and 540nm by using an enzyme-labeled instrument. The OD value read at 450nm is subtracted from the OD value read at 540nm, and the value is taken as an ordinate, the TF/FVIIa standard concentration is taken as an abscissa, a standard curve is drawn, and the TF/FVIIa content of the sample can be converted into the corresponding concentration from the standard curve according to the OD value. The differences in TF/FVIIa levels of each group were analyzed by the method of one-way analysis of variance (ANOVA one-way, LSD) in SPSS22.0, and the plasma TF/FVIIa levels are shown in Table 6.
TABLE 6 treatment of TF/FVIIa levels in rat plasma
a) P < 0.01. N=4, compared with normal saline
The data in Table 6 show that compound 6 was effective in reducing the plasma TF/FVIIa levels in rats at a 0.82. Mu. Mol/kg oral dose for 4 days following oral administration. It can be seen that the invention has remarkable technical effects.
EXAMPLE 19 evaluation of Gly-Pro-Arg-Pro-NHCH 2 CH 2 Effect of NH-warfarin (6) on rat plasma Soluble Fibrin Monomer Complex (SFMC) content
Whole blood of rats collected in example 13, 4.0 mL/mouse, was placed in a centrifuge tube containing 0.4mL of sodium citrate solution (3.8%), and after centrifugation at 1000g for 15 minutes, upper layer Platelet Poor Plasma (PPP) was aspirated. After diluting PPP samples 200 times by using sample diluent in the kit, adding 100 mu L of each sample and standard substances into each hole, covering a cover plate film, incubating at 37 ℃ for 2 hours, pouring all liquid in the plate, directly adding 100 mu L of Biotin solution into each hole without washing the plate, covering a new cover plate film, and incubating at 37 ℃ for 1 hour. The plate was then discarded, 200. Mu.L of wash solution was added to each well, and after 2min of standing, the wash solution was discarded and the 96-well plate was dried by pipetting, and this was repeated 3 times. Next, 100. Mu.L of HRP-avidin was added to each well and covered with a fresh cover film and incubated at 37℃for 1 hour. After the incubation, the plate washing operation was repeated 5 times to remove the liquid in each well. After adding 90. Mu.L of 3,3', 5' -tetramethylbenzidine to each well, covering a new cover plate film, incubating for 20 minutes at 37 ℃ in a dark place, adding 50. Mu.L of stop solution to each well, and rapidly reading the OD value of each well at two wavelengths of 450nm and 540nm by using an enzyme-labeled instrument. The OD value read at 450nm is subtracted from the OD value read at 540nm, and the value is taken as an ordinate, the concentration of the SFMC standard substance is taken as an abscissa, a standard curve is drawn, and the SFMC content of the sample can be converted into the corresponding concentration from the standard curve according to the OD value. The difference in SFMC content of each group was analyzed by the one-way analysis of variance (LSD) method in SPSS22.0, and the SFMC content in rat plasma was as shown in Table 7.
TABLE 7 treatment of SFMC content in rat plasma
a) P is less than 0.05 compared with normal saline; n=5
The data in Table 7 show that compound 6 was effective in reducing the level of SFMC in rat plasma at an oral dose of 0.82. Mu. Mol/kg for 4 days following oral administration. It can be seen that the invention has remarkable technical effects.
Claims (6)
1.Gly-Pro-Arg-Pro-NHCH 2 CH 2 NH-warfarin has the structural formula:
2. a process for preparing Gly-Pro-Arg-Pro-NHCH as claimed in claim 1 2 CH 2 A method of NH-warfarin, the method comprising the steps of:
1) Synthesizing warfarin-4-O-benzyl acetate;
2) Synthesizing warfarin-4-O-acetic acid;
3) Synthesis of Boc-NHCH 2 CH 2 NH-warfarin;
4) Boc-NHCH 2 CH 2 Conversion of NH-warfarin to HCl NH 2 CH 2 CH 2 NH-warfarin;
5) Synthesis of Boc-Gly-Pro-Arg (NO) 2 )-Pro-OBzl；
6) Boc-Gly-Pro-Arg (NO) 2 ) Conversion of Pro-OBzl to Boc-Gly-Pro-Arg (NO) 2 )-Pro；
7) HCl. NH 2 CH 2 CH 2 NH-warfarin and Boc-Gly-Pro-Arg (NO) 2 ) Synthesis of Boc-Gly-Pro-Arg (NO) by Pro reaction 2 )-Pro-NHCH 2 CH 2 NH-warfarin;
8) Boc-Gly-Pro-Arg (NO) 2 )-Pro-NHCH 2 CH 2 Conversion of NH-warfarin to Gly-Pro-Arg-Pro-NHCH according to claim 1 2 CH 2 NH-warfarin.
3. The Gly-Pro-Arg-Pro-NHCH of claim 1 2 CH 2 Application of NH-warfarin in preparing antithrombotic medicine without hemorrhage side effect is provided.
4. The Gly-Pro-Arg-Pro-NHCH of claim 1 2 CH 2 Use of NH-warfarin in the preparation of a clotting factor II antagonist.
5. The Gly-Pro-Arg-Pro-NHCH of claim 1 2 CH 2 Use of NH-warfarin in the preparation of a tissue factor/factor VII antagonist.
6. The Gly-Pro-Arg-Pro-NHCH of claim 1 2 CH 2 Use of NH-warfarin in the preparation of a soluble fibrin monomer complex antagonist.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| LV14606A (en) * | 2011-05-17 | 2012-11-20 | Tetra, Sia | Novel factor xii inhibitor |
| CN107686506A (en) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | The O acetyl GPRP of warfarin 4, it is synthesized, activity and application |
| RU2645414C1 (en) * | 2016-10-06 | 2018-02-21 | Общество с ограниченной ответственностью фирма "Технология-Стандарт" | Method for preventing bleeding caused by use of warfarin in experiment |
| CN110711191A (en) * | 2018-07-12 | 2020-01-21 | 北京恒润泰生医药科技有限公司 | Molecular complex of warfarin and vitamin C, preparation, activity and application thereof |
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| US20040235942A1 (en) * | 2003-01-30 | 2004-11-25 | Ridker Paul M. | Method for preventing recurring venous thromboembolism using long-term low-intensity warfarin |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| LV14606A (en) * | 2011-05-17 | 2012-11-20 | Tetra, Sia | Novel factor xii inhibitor |
| CN107686506A (en) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | The O acetyl GPRP of warfarin 4, it is synthesized, activity and application |
| RU2645414C1 (en) * | 2016-10-06 | 2018-02-21 | Общество с ограниченной ответственностью фирма "Технология-Стандарт" | Method for preventing bleeding caused by use of warfarin in experiment |
| CN110711191A (en) * | 2018-07-12 | 2020-01-21 | 北京恒润泰生医药科技有限公司 | Molecular complex of warfarin and vitamin C, preparation, activity and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Synthesis of quinazolinone conjugated shorter analogues of Bactenecin7 as potent antimicrobials;R. Suhas等;Protein & Peptide Letters;第20卷(第2期);146-155 * |
| 华法林的神奇之旅;许俊堂;;临床误诊误治(第10期);117 * |
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