CN108976285A - The warfarin of Gly-Pro-Arg-Pro-AA modification, synthesis, activity and application - Google Patents

The warfarin of Gly-Pro-Arg-Pro-AA modification, synthesis, activity and application Download PDF

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CN108976285A
CN108976285A CN201710414648.7A CN201710414648A CN108976285A CN 108976285 A CN108976285 A CN 108976285A CN 201710414648 A CN201710414648 A CN 201710414648A CN 108976285 A CN108976285 A CN 108976285A
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gly
warfarin
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obzl
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CN108976285B (en
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赵明
彭师奇
王玉记
吴建辉
侯梦雨
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Capital Medical University
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Abstract

The invention discloses the warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA of following formula, (AA is Ala residue in formula, Glu residue, Gly residue and Pro residue), disclose their preparation method, their anti-phlebothrombosis activity is disclosed, thus the invention discloses them to prepare the application in anti-phlebothrombosis drug.

Description

The warfarin of Gly-Pro-Arg-Pro-AA modification, synthesis, activity and application
Technical field
The present invention relates to warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA (in formula AA be Ala residue, Glu residue, Gly residue and Pro residue), it is related to their preparation method, the anti-phlebothrombosis for being related to them is active, thus the present invention relates to They are preparing the application in anti-phlebothrombosis drug.The invention belongs to biomedicine fields.
Background technique
Venous Thrombosis has become disease incidence height instantly and the high disease of the death rate.Venous Thrombosis mainly includes Deep venou Thrombus and pulmonary embolism.Deep vein thrombosis and patient's number of pulmonary embolism morbidity have not exceeded only total people of myocardial infarction and apoplexy morbidity Number, and the total number of persons more dead than caused by breast cancer and AIDS is high.Since the disease incidence of Venous Thrombosis increases with the age Exponentially state increases, so especially serious to the threat of the people's health of aging country as China.If population base Number is taken into account, especially serious to the absolute negative effect of China's national economy.Then, the prevention and treatment one of Venous Thrombosis It is directly field of medicaments emphasis of interest.Although warfarin conduct represents drug and is used for clinical practice in nineteen forty-one, magnificent The security window of method woods is narrow.Underdosage will lead to pulmonary embolism, the excessively high risk for having fatal hemorrhage of dosage.Past more than 50 years A large amount of structure of modification have been carried out to warfarin, all can not obtain the satisfied analog of anti-phlebothrombosis activity but.With tradition Thinking it is different, the target that inventor modifies warfarin structure is the security window for expanding warfarin.Make them under low dosage The satisfied anti-phlebothrombosis activity of display avoids pulmonary embolism risk, their high dose is made to remain on the conventional agent lower than warfarin Amount avoids the risk of bleeding.It was explored by 5 years, inventor has found that 4 of warfarin are repaired with acetyl Gly-Pro-Arg-Pro-AA Warfarin -4-O- acetyl-the Gly-Pro-Arg- that decorations (AA is Ala residue, Glu residue, Gly residue and Pro residue) generate Pro-AA is under 0.1 μm of ol/kg dosage with outstanding anti-phlebothrombosis activity.4.87 μ of dosage of this dose ratio warfarin Mol/kg is 48.7 times low, that is to say, that their security window is 48.7 times bigger than the security window of warfarin.As it can be seen that the present invention has There is significant technological progress.According to this discovery, the present invention is inventors herein proposed.
Summary of the invention
First content of the invention is to provide the warfarin -4-O- second that first content of the invention is to provide following formula Acyl-Gly-Pro-Arg-Pro-AA (AA is Ala residue, Glu residue, Gly residue and Pro residue in formula).
Second content of the invention is to provide warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA, and (AA is in formula Ala residue, Glu residue, Gly residue and Pro residue) preparation method, this method comprises:
1) warfarin -4-O- benzyl acetate is synthesized；
2) warfarin -4-O- acetic acid is synthesized；
3) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, is synthesized using liquid phase method Gly-Pro-OBzl；
3) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes China using liquid phase method Fa Lin -4-O- acetyl-Gly-Pro；
4) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes Arg using liquid phase method (NO2)-Pro-Ala-OBzl,Arg(NO2)-Pro-Glu(OBzl)-OBzl,Arg(NO2)-Pro-Gly-OBzl,Arg(NO2)- Pro-Pro-OBzl；
5) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes China using liquid phase method Fa Lin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Ala-OBzl, warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)- Pro-Glu (OBzl)-OBzl, warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Gly-OBzl, warfarin -4-O- Acetyl-Gly-Pro-Arg (NO2)-Pro-Pro-OBzl；
6) hydrogenolysis Deprotection synthesizes warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Ala, warfarin -4-O- second Acyl-Gly-Pro-Arg-Pro-Glu, warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Gly, warfarin -4-O- acetyl - Gly-Pro-Arg-Pro-Pro。
Third content of the invention is that (AA is evaluation warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA in formula Ala residue, Glu residue, Gly residue and Pro residue) anti-phlebothrombosis activity.
Detailed description of the invention
The bromo- 2- benzyl acetate of synthetic route (i) of Fig. 1 warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA, third Ketone, K2CO3,45℃；(ii)CH3OH,Pd/C,H2；(iii) dicyclohexylcarbodiimide (DCC), I-hydroxybenzotriazole (HOBt), N-methylmorpholine (NMM), tetrahydrofuran (THF)；(iv) ethyl acetate solution (4N) of hydrogen chloride；AA in 5a and 6a For Ala residue；AA is Glu residue in 5b and 6b；AA is Gly residue in 5c and 6c；AA is Pro residue in 5d and 6d.
Specific embodiment
In order to which the present invention is further explained, a series of embodiments are given below.These embodiments be entirely it is illustrative, it Only be used to the present invention is specifically described, be not construed as limitation of the present invention.
Embodiment 1 prepares warfarin -4-O- benzyl acetate
26.48g (80.00mmol) warfarin and 400mL acetone are dissolved in 45 DEG C of stirrings to warfarin.It is molten toward obtaining In liquid plus 14mL (88mmol) benzyl acetate bromide, 45 DEG C of stirring 96h are continued at, thin-layer chromatography (TLC, petrol ether/ethyl acetate= 2/1) display reaction is completed.Reaction solution, which is crossed, to be filtered out, and filtrate decompression concentration, obtained light yellow oil is purified with silica gel column chromatography (petrol ether/ethyl acetate=8/1) obtains 19.77g (54%) title compound, is colorless 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).
Embodiment 2 prepares warfarin -4-O- acetic acid
19.77g (43.36mmol) warfarin -4-O- benzyl acetate is dissolved in 150mL tetrahydrofuran, then with 4.94g palladium Carbon (Pd/C) suspends.Lead to hydrogen 72h inward.TLC (petrol ether/ethyl acetate=2/1) shows fully reacting.Filter off Pd/C, filter Liquid is concentrated under reduced pressure, and obtains 15.58g (98%) title compound, is 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(dd,J1=15.0Hz, J2=30.0Hz, 2H), 3.54~3.47 (m, 2H), 2.14 (s, 3H).
Embodiment 3 prepares Boc-Gly-Pro-OBzl
7.730g (44.15mmol) Boc-Gly 100mL anhydrous tetrahydro furan is dissolved, 0 DEG C adds into the solution 5.940g (44.00mmol) I-hydroxybenzotriazole (HOBt) and 9.888g (48.00mmol) dicyclohexylcarbodiimide (DCC), 0 DEG C of stirring 30min and inward plus 9.665g (40.00mmol) HClPro-OBzl.Reaction solution N-methylmorpholine tune Saving pH is 9, and 17h is stirred at room temperature, and TLC (methylene chloride/methanol=20/1) display reaction is completed.It filters out insoluble in reaction solution Object, filtrate decompression concentration, residue 150mL ethyl acetate dissolve, and filter off colorless solid, filtrate is respectively with saturation NaHCO3It is molten Liquid is washed (50mL × 3), and saturation NaCl solution washes (50mL × 3), 5%KHSO4Solution is washed (50mL × 3), and saturation NaCl solution is washed (50mL × 3) are saturated NaHCO3Solution is washed (50mL × 3), and saturation NaCl solution is washed (50mL × 3), and ethyl acetate phase is with anhydrous Sodium sulphate dries 12h, filtering, filtrate decompression concentration, and obtained faint yellow solid silica gel column chromatography purifies (petroleum ether/acetic acid Ethyl ester=3/1), 13.42g (93%) title compound is obtained, is colorless solid.ESI-MS(m/e):363[M+H]+。
Embodiment 4 prepares HClGly-Pro-OBzl
The ethyl acetate solution (4M) of 13.42g (37.05mmol) Boc-Gly-Pro-OBzl and 52mL hydrogen chloride is slow Mixing.Obtained solution stirs 5h in ice bath.Later, reaction mixture is concentrated under reduced pressure.Residue 50mL anhydrous acetic acid second Ester dissolution, obtained solution are concentrated under reduced pressure.The operation is in triplicate.Residue is sufficiently washed with anhydrous ether, obtains 10.85g (98%) title compound is colorless solid.
Embodiment 5 prepares warfarin -4-O- acetyl-Gly-Pro-OBzl
According to the method for embodiment 3, from 15.58g (42.57mmol) warfarin -4-O- acetic acid and 11.56g (38.70mmol) HClPro-OBzl obtains 21.07g (89%) title compound, is colorless solid.ESI-MS(m/e):611 [M+H]+；1H-NMR(300MHz,DMSO-d6) δ/ppm=8.50 (dd, J1=5.7Hz, J2=11.4Hz, 1H), 7.91 (dt, J1 =1.5Hz, J2=8.7Hz, 1H), 7.64 (td, J1=1.5Hz, J2=6.0Hz, 1H), 7.43 (d, J=7.5Hz, 4H), 7.34 (m, 6H), 7.29 (t, J=3.3Hz, 2H), 7.19 (m, 1H), 5.15 (m, 2H), 4.92 (t, J=7.5Hz, 1H), 4.58 (t, J =3.9Hz, 2H), 4.44 (dd, J1=3.9Hz, J2=9.0Hz, 1H), 4.10 (m, 2H), 3.61 (m, 2H), 3.47 (d, J= 7.2Hz, 2H), 2.22 (m, 1H), 2.14 (s, 3H), 1.99~1.76 (m, 3H).
Embodiment 6 prepares warfarin -4-O- acetyl-Gly-Pro
According to the method for embodiment 2, obtained from 21.07g (34.53mmol) warfarin -4-O- acetyl-Gly-Pro-OBzl 14.68g (82%) title compound is colorless solid.ESI-MS(m/e):521[M+H]+。
Embodiment 7 prepares Boc-Pro-Ala-OBzl
According to the method for embodiment 3, from 1.659g (7.716mmol) Boc-Pro and 2.456g (6.995mmol) Tos Ala-OBzl obtains 2.184g (83%) title compound, is faint yellow solid.ESI-MS(m/e):377[M+H]+。
Embodiment 8 prepares HClPro-Ala-OBzl
According to the method for embodiment 4,1.571g is obtained from 1.890g (5.025mmol) Boc-Pro-Ala-OBzl (100%) title compound is yellow oil.
Embodiment 9 prepares Boc-Arg (NO2)-Pro-Ala-OBzl
According to the method for embodiment 3, from 1.749g (5.483mmol) Boc-Arg (NO2) and 1.563g (5.000mmol) HClPro-Ala-OBzl obtains 1.964g (68%) title compound, is colorless solid.1H-NMR(300MHz,DMSO-d6) δ/ppm=8.50 (m, 1H), 8.32 (d, J=6.9Hz, 1H), 7.36 (s, 5H), 6.91 (d, J=7.5Hz, 1H), 5.10 (dd, J1=12.3Hz, J2=21.6Hz, 2H), 4.35~4.31 (m, 1H), 4.30 (q, J=7.2Hz, 1H), 4.15 (m, 1H), 3.62 ~3.53 (m, 2H), 3.13 (s, 2H), 2.03~1.45 (m, 8H), 1.37 (s, 9H), 1.30 (d, J=7.2Hz, 3H).
Embodiment 10 prepares HClArg (NO2)-Pro-Ala-OBzl
According to the method for embodiment 4, from 1.072g (1.857mmol) Boc-Arg (NO2)-Pro-Ala-OBzl obtains 0.912g (96%) title compound is colorless solid.
Embodiment 11 prepares warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Ala-OBzl(5a)
According to the method for embodiment 3, from 1.027g (1.974mmol) warfarin -4-O- acetyl-Gly-Pro and 0.859g (1.80mmol) HClPro-Ala-OBzl obtains 0.580g (35%) title compound, is colorless solid1H-NMR (300MHz,DMSO-d6) δ/ppm=8.59~8.41 (m, 2H), 8.32 (m, 1H), 8.14 (d, J=7.5Hz, 1H), 7.90 (d, J=7.5Hz, 1H), 7.64 (t, J=7.5Hz, 1H), 7.43 (d, J=8.7Hz, 2H), 7.36 (s, 7H), 7.27 (t, J= 7.5Hz, 2H), 7.18 (t, J=6.9Hz, 1H), 5.10 (dd, J1=12.3Hz, J2=21.0Hz, 2H), 4.91 (t, J= 6.9Hz, 1H), 4.58 (s, 2H), 4.52~4.22 (m, 2H), 4.31~4.27 (m, 2H), 4.07 (m, 2H), 3.57~3.45 (m, 4H), 3.46 (d, J=6.9Hz, 2H), 3.14 (s, 2H), 2.14 (s, 3H), 2.07~1.56 (m, 12H), 1.34 (d, J= 12.6Hz,3H)。
Embodiment 12 prepares warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Ala (6a)
According to the method for embodiment 2, from 0.200g (0.204mmol) warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Ala-OBzl (5a) obtains 0.042g (24%) title compound, it is colorless solid.ESI-MS(m/e):845[M +H]+,867[M+Na]+；1H-NMR(300MHz,DMSO-d6): δ/ppm=9.64 (m, 1H), 8.43 (d, J=5.1Hz, 1H), 8.25 (d, J=6.9Hz, 1H), 7.90 (d, J=7.8Hz, 1H), 7.64 (t, J=7.2Hz, 1H), 7.43 (d, J=8.4Hz, 2H), 7.36 (d, J=7.5Hz, 2H), 7.29 (s, 1H), 7.26 (t, J=7.2Hz, 2H), 7.19 (m, 3H), 4.90 (t, J= 7.2Hz, 1H), 4.58 (d, J=2.7Hz, 2H), 4.50 (m, 1H), 4.40 (dd, J1=2.7Hz, J2=8.1Hz, 1H), 4.17 (m, 1H), 4.01 (m, 2H), 3.72 (m, 2H), 3.64~3.50 (m, 3H), 3.46 (d, J=7.2Hz, 2H), 3.10 (s, 2H), 2.14 (s, 3H), 2.11~1.54 (m, 12H), 1.15 (d, J=6.6Hz, 3H).
Embodiment 13 prepares Boc-Pro-Glu (OBzl)-OBzl
According to the method for embodiment 3, from 1.659g (7.716mmol) Boc-Pro and 2.547g (7.004mmol) HCl Glu (OBzl)-OBzl obtains 3.484g (95%) title compound, is light yellow oil.ESI-MS(m/e):525[M+H ]+。
Embodiment 14 prepares HClPro-Glu (OBzl)-OBzl
According to the method for embodiment 4,3.062g is obtained from 3.484g (6.647mmol) Boc-Pro-Glu (OBzl)-OBzl (100%) title compound is yellow oil.
Embodiment 15 prepares Boc-Arg (NO2)-Pro-Glu(OBzl)-OBzl
According to the method for embodiment 3, from 2.456g (7.699mmol) Boc-Arg (NO2) and 3.062g (6.647mmol) HClPro-Glu (OBzl)-OBzl obtains 2.173g (45%) title compound, is colorless solid1H-NMR(300MHz, DMSO-d6) δ/ppm=8.33 (d, J=7.5Hz, 1H), 7.36 (s, 10H), 6.92 (d, J=7.5Hz, 1H), 5.15~5.05 (m,4H),4.34(m,2H),4.15(m,1H),3.58(m,2H),3.10(s,2H),2.47(m,2H),2.03(m.2H),1.94 ~1.45 (m, 8H), 1.37 (s, 9H).
Embodiment 16 prepares HClArg (NO2)-Pro-Glu(OBzl)-OBzl
According to the method for embodiment 4, from 1.816g (2.504mmol) Boc-Arg (NO2)-Pro-Glu(OBzl)-OBzl 1.621g (98%) title compound is obtained, is colorless oil.
Embodiment 17 prepares warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Glu(OBzl)-OBzl(5b)
According to the method for embodiment 3, from 1.427g (2.743mmol) warfarin -4-O- acetyl-Gly-Pro and 1.621g (2.450mmol)HCl·Arg(NO2)-Pro-Glu (OBzl)-OBzl obtains 0.539g (20%) title compound, it is colourless Solid1H-NMR(300MHz,DMSO-d6) δ/ppm=8.48~8.45 (m, 2H), 8.36 (t, J=7.2Hz, 1H), 8.19 (d, J=7.8Hz, 1H), 7.89 (d, J=7.2Hz, 1H), 7.63 (t, J=8.1Hz, 1H), 7.42 (d, J=8.4Hz, 1H), 7.35 (s, 13H), 7.26 (t, J=7.5Hz, 2H), 7.17 (d, J=6.9Hz, 1H), 5.14~5.04 (m, 4H), 4.90 (t, J= 7.2Hz, 1H), 4.57 (s, 2H), 4.50~4.31 (m, 4H), 4.05 (m, 2H), 3.64~3.44 (m, 4H), 3.46 (d, J= 7.2Hz, 2H), 3.11 (s, 2H), 2.46 (m, 2H), 2.14 (s, 3H), 2.05~1.54 (m, 14H).
Embodiment 18 prepares warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Glu (6b)
According to the method for embodiment 2, from 0.200g (0.177mmol) warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Glu (OBzl)-OBzl (5b) obtains 0.015g (9%) title compound, it is colorless solid.ESI-MS(m/e): 903[M+H]+；1H-NMR(300MHz,D2O): δ/ppm=7.48~7.42 (m, 2H), 7.21~7.11 (m, 7H), 4.78 (t, J =7.2Hz, 1H), 4.53~4.27 (m, 4H), 4.14 (s, 2H), 4.12 (m, 2H), 3.62~3.30 (m, 6H), 2.99 (s, 2H), 2.27 (t, J=7.8Hz, 2H), 2.11 (s, 3H), 2.05~1.55 (m, 14H).
Embodiment 19 prepares Boc-Pro-Gly-OBzl
According to the method for embodiment 3, from 1.423g (6.619mmol) Boc-Pro and 1.208g (5.995mmol) HCl Gly-OBzl obtains 2.079g (96%) title compound, is light yellow oil.ESI-MS(m/e):363[M+H]+。
Embodiment 20 prepares HClPro-Gly-OBzl
According to the method for embodiment 4,1.234g (100%) is obtained from 1.50g (4.14mmol) Boc-Pro-Gly-OBzl Title compound is yellow oil.
Embodiment 21 prepares Boc-Arg (NO2)-Pro-Gly-OBzl
According to the method for embodiment 3, from 1.754g (5.498mmol) Boc-Arg (NO2) and 1.492g (5.000mmol) HClPro-Gly-OBzl obtains 0.987g (35%) title compound, is colorless solid1H-NMR(300MHz,DMSO-d6) δ/ppm=8.24 (t, J=6.0Hz, 1H), 7.36 (m, 5H), 6.93 (d, J=7.8Hz, 1H), 5.12 (s, 2H), 4.34 (dd, J1=3.3Hz, J2=8.1Hz, 1H), 4.17 (m, 1H), 3.93 (dd, J1=6.0Hz, J2=17.4Hz, 1H), 3.84 (dd, J1 =5.7Hz, J2=17.4Hz, 1H), 3.59 (m, 2H), 3.13 (s, 2H), 2.04~1.40 (m, 8H), 1.37 (s, 9H).
Embodiment 22 prepares HClArg (NO2)-Pro-Gly-OBzl
According to the method for embodiment 4, from 0.957g (1.70mmol) Boc-Arg (NO2)-Pro-Gly-OBzl obtains 0.828g (98%) title compound is colorless solid.
Embodiment 23 prepares warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Gly-OBzl(5c)
According to the method for embodiment 3, from 0.973g (1.87mmol) warfarin -4-O- acetyl-Gly-Pro and 0.828g (1.66mmol)HCl·Arg(NO2)-Pro-Gly-OBzl obtains 0.545g (34%) title compound, it is colorless solid1H- NMR(300MHz,DMSO-d6) δ/ppm=8.43~8.38 (m, 2H), 8.27 (m, 1H), 8.17 (d, J=7.2Hz, 1H), 7.90 (d, J=7.2Hz, 1H), 7.64 (t, J=8.1Hz, 1H), 7.43 (d, J=8.7Hz, 1H), 7.37 (s, 8H), 7.26 (t, J=7.2Hz, 2H), 7.18 (d, J=6.9Hz, 1H), 5.12 (s, 2H), 4.91 (t, J=7.2Hz, 1H), 4.58 (s, 2H), 4.49 (m, 1H), 4.40 (m, 1H), 4.33 (m, 1H), 4.05 (m, 2H), 3.88 (m, 2H), 3.69~3.48 (m, 4H), 3.46 (d, J=7.2Hz, 2H), 3.14 (s, 2H), 2.14 (s, 3H), 2.08~1.50 (m, 12H).
Embodiment 24 prepares warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Gly (6c)
According to the method for embodiment 2, from 0.200g (0.207mmol) warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Gly-OBzl (5c) obtains 0.037g (22%) title compound, it is colorless solid.ESI-MS(m/e):831[M +H]+,853[M+Na]+,869[M+K]+；1H-NMR(300MHz,D2O): δ/ppm=7.31~6.92 (m, 9H), 4.59 (m, 1H), 4.49~4.27 (m, 3H), 4.17~4.07 (m, 3H), 3.61 (d, J=4.8Hz, 2H), 3.67~3.32 (m, 6H), 2.97 (s, 2H), 2.04 (s, 3H), 2.21~1.52 (m, 12H).
Embodiment 25 prepares Boc-Pro-Ile-OBzl
According to the method for embodiment 3, from 1.420g (6.605mmol) Boc-Pro and 1.817g (4.621mmol) Tos Ile-OBzl obtains 1.921g (99%) title compound, is light yellow oil.ESI-MS(m/e):419[M+H]+。
Embodiment 26 prepares Boc-Pro-Pro-OBzl
According to the method for embodiment 2, from 1.420g (6.605mmol) Boc-Pro and 1.448g (5.993mmol) HCl Pro-OBzl obtains 2.234g (93%) title compound, is light yellow oil.ESI-MS(m/e):403[M+H]+。
Embodiment 27 prepares HClPro-Pro-OBzl
According to the method for embodiment 4,1.512g is obtained from 1.796g (4.466mmol) Boc-Pro-Pro-OBzl (100%) title compound is faint yellow solid.
Embodiment 28 prepares Boc-Arg (NO2)-Pro-Pro-OBzl
According to the method for embodiment 3, from 1.858g (5.824mmol) Boc-Arg (NO2) and 1.512g (4.465mmol) HClPro-Pro-OBzl obtains 1.679g (62%) title compound, is colorless solid1H-NMR(300MHz,DMSO-d6) δ/ppm=7.36 (s, 5H), 6.89 (d, J=7.5Hz, 1H), 5.09 (dd, J1=12.6Hz, J2=29.7Hz, 2H), 4.60 (dd,J1=3.6Hz, J2=8.1Hz, 1H), 4.37 (dd, J1=3.9Hz, J2=8.7Hz, 1H), 4.17 (m, 1H), 3.68 (m, 2H), 3.51 (m, 2H), 3.13 (s, 2H), 2.14 (m, 2H), 1.97~1.85 (m, 5H), 1.75~1.50 (m, 5H), 1.37 (s,9H)。
Embodiment 29 prepares HClArg (NO2)-Pro-Pro-OBzl
According to the method for embodiment 4, from 1.472g (2.441mmol) Boc-Arg (NO2)-Pro-Pro-OBzl obtains 1.317g (100%) title compound is colorless solid.
Embodiment 30 prepares warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Pro-OBzl(5d)
According to the method for embodiment 3, from 1.397g (2.686mmol) warfarin -4-O- acetyl-Gly-Pro and 1.317g (2.441mmol)HCl·Arg(NO2)-Pro-Pro-OBzl obtains 0.892g (36%) title compound, it is colorless solid1H- NMR(300MHz,DMSO-d6) δ/ppm=8.43~8.38 (m, 2H), 8.12 (d, J=7.5Hz, 1H), 7.90 (d, J= 7.5Hz, 1H), 7.64 (t, J=7.5Hz, 1H), 7.43 (d, J=8.7Hz, 2H), 7.35 (s, 7H), 7.26 (t, J=7.2Hz, 2H), 7.18 (d, J=7.2Hz, 1H), 5.09 (dd, J1=12.6Hz, J2=29.7Hz, 2H), 4.91 (t, J=7.2Hz, 1H), 4.58 (s, 2H), 4.60~4.54 (m, 2H), 4.45~4.35 (m, 2H), 4.06 (m, 2H), 3.65~3.48 (m, 6H), 3.46 (d, J=7.2Hz, 2H), 3.14 (s, 2H), 2.14 (s, 3H), 2.17~1.60 (m, 16H).
Embodiment 31 prepares warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Pro (6d)
According to the method for embodiment 2, from 0.200g (0.199mmol) warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Pro-OBzl (5d) obtains 0.045g (26%) title compound, it is colorless solid.ESI-MS(m/e):871[M +H]+,893[M+Na]+,909[M+K]+；1H-NMR(300MHz,DMSO-d6): δ/ppm=8.43 (m, 1H), 7.90 (m, 1H), 7.63 (t, J=8.1Hz, 1H), 7.42 (d, J=8.1Hz, 2H), 7.39~7.35 (m, 2H), 7.26 (t, J=7.2Hz, 2H), 7.17 (m, 1H), 6.95 (s, 1H), 4.89 (m, 1H), 4.59 (d, J=6.6Hz, 2H), 4.56~4.47 (m, 3H), 4.20~ 3.90 (m, 3H), 3.46 (d, J=7.2Hz, 2H), 3.52~3.50 (m, 6H), 2.54 (s, 2H), 2.27 (m, 2H), 2.14 (s, 3H), 2.10~1.39 (m, 14H).
Embodiment 32 evaluates the anti-phlebothrombosis effect of 6a-d
20% urethane of reagent (urethanes, CAS:51-79-6, Sinopharm Chemical Reagent Co., Ltd.) is raw Reason saline solution is anesthetic, physiological saline (Shijiazhuang Siyao Co., Ltd), warfarin sodium (CAS:129-06-6, lark prestige Science and Technology Ltd.).
Animal SD strain rats, male, 300 ± 20g are purchased from Beijing Vital River Experimental Animals Technology Co., Ltd..
Rat model Ligation of inferior vena cava model.
The oral dose of dosage the compound of the present invention 6a-d is 0.1 μm of ol/kg, the mouth of positive control warfarin Taking dosage is 4.87 μm of ol/kg, and negative control is physiological saline.
Experimental implementation rat adapts to environment and fasting one day before surgery, by dosage above to rat oral gavage.Administration Make rat anesthesia in 20% urethane solution intraperitoneal administration of operation consent 2min after 30min.Anesthetized rat is fixed on rat to consolidate On fixed board, then abdomen preserved skin and disinfection open abdominal cavity along hunter's line, and up to exposing one jiao of liver, be open about 4cm long.It removes The organs such as intraperitoneal small intestine are simultaneously wrapped up with the gauze for infiltrating physiological saline.Blunt separation blood vessel surrounding connective tissue, under exposure Vena cave and its branch, peel away abdominal aorta and inferior caval vein below left renal vein, are then soaked with physiological saline The organs such as intestines, by Ligation of inferior vena cava, are moved back to abdominal cavity by anatomical position in inferior caval vein and left renal vein intersection by suture, With suture layer-by-layer suture abdominal cavity.Postoperative, rat recycles 4 hours in 25 DEG C.After opening abdominal cavity, branch is ligatured one by one, from cavity of resorption 2cm inferior caval vein is taken out at the ligation of the intersection of vein and left renal vein, is taken out thrombus.Thrombus is weighed and is examined with t Statistical result.Operation with every group four only alternately.Experimental data is shown in Table 1.
Influence of the 1 compound 6a-d of table to Ligation of inferior vena cava rat vein thrombus weight
With left renal vein intersection by Ligation of inferior vena cava, the organs such as intestines are moved back to abdominal cavity by anatomical position, use suture Layer-by-layer suture abdominal cavity.
It recycles 4 hours in the postoperative environment that rat is placed in 25~28 DEG C, after opening abdominal cavity, one by one ligatures its branch, 2cm inferior caval vein is taken out since ligation place of the intersection of inferior caval vein and left renal vein, is taken out thrombus.Thrombus weighing And the statistical result in the way of t inspection.Operation with every group four only alternately.Experimental data is shown in Table 1.
N=8
A) compared with physiological saline, p < 0.05
B) compared with physiological saline, p<0.05, compared with Warfarin, p>0.05
Experimental result:
The result shows that 0.1 μm of ol/kg dosage 6a, 6c, 6d and 6h treatment rat bolt weight with 4.87 μm of ol/kg agent The bolt heavy phase of the rat of warfarin treatment is measured as (10.9 ± 4.9mg, p > 0.05).As it can be seen that the compound of the present invention 6a, 6c, 6d And the anti-phlebothrombosis activity of 6h is at least 48.7 times stronger than warfarin.
Embodiment 33 evaluates the anti-phlebothrombosis effect of 6b, 6e, 6f, 6g, 6i and 6j
Experimental material
Urethane (urethanes, CAS:51-79-6, Sinopharm Chemical Reagent Co., Ltd.), physiological saline (stone Four medicine Co., Ltd of the village of family) warfarin sodium (CAS:129-06-6, lark prestige Science and Technology Ltd.).
Experimental animal
SD strain rats, male, 300 ± 20g are purchased from Beijing Vital River Experimental Animals Technology Co., Ltd..
Experimental method
Experiment uses rat Ligation of inferior vena cava model.
Grouping and dosage:
The compound of the present invention 6b, 6e, 6f, 6g, 6i and 6j dosage is 0.1 μm of ol/kg, the dosage of positive control warfarin For 4.87 μm of ol/kg, negative control is physiological saline.
Preparation of reagents
Anesthetic is 20% urethane solution of normal saline.
Experimental implementation:
Rat adapts to environment and fasting one day before surgery, carries out stomach-filling to rat with the dosage of 0.3mL/100g weight and gives Medicine.It is anaesthetized after administration 30min in 20% urethane solution intraperitoneal administration of operation consent 2min.Rat is fixed on rat fixed plate On, then abdomen preserved skin and disinfection open abdominal cavity along hunter's line, up to one jiao of liver of exposing, openings of sizes about 4cm long. It removes the organs such as intraperitoneal small intestine and is wrapped up with the gauze for infiltrating physiological saline.Blunt separation blood vessel surrounding connective tissue, cruelly Reveal inferior caval vein and its branch, peel away abdominal aorta and inferior caval vein below left renal vein, then uses physiology salt water logging The organs such as intestines, by Ligation of inferior vena cava, are moved back to abdomen by anatomical position in inferior caval vein and left renal vein intersection by wet suture Chamber, with suture layer-by-layer suture abdominal cavity.
It recycles 4 hours in the postoperative environment that rat is placed in 25~28 DEG C, after opening abdominal cavity, one by one ligatures its branch, 2cm inferior caval vein is taken out since ligation place of the intersection of inferior caval vein and left renal vein, is taken out thrombus.Thrombus weighing And the statistical result in the way of t inspection.Operation with every group four only alternately.Experimental data is shown in Table 2.
Table 2
N=7
A) compared with physiological saline, p < 0.05
B) compared with physiological saline, p<0.05, compared with Warfarin, p>0.05
C) compared with physiological saline, p<0.01, compared with Warfarin, p>0.05
D) compared with physiological saline, p < 0.01, compared with Warfarin, p < 0.05
Experimental result:
The result shows that 0.1 μm of ol/kg dosage 6e, 6g, 6i and 6j treatment rat bolt weight with 4.87 μm of ol/kg agent The bolt heavy phase of the rat of warfarin treatment is measured as (21.3 ± 4.7mg, p > 0.05), and 0.1 μm of ol/kg dosage 6b and 6f are treated Rat significantly less than 4.87 μm ol/kg dosage warfarin treatments of bolt weight rat bolt weight (21.3 ± 4.7mg, p < 0.05), it is seen then that the anti-phlebothrombosis activity of the compound of the present invention 6b, 6e, 6f, 6g, 6i and 6j are at least stronger by 48.7 than warfarin Times.

Claims (3)

1. warfarin -4-O- acetyl-the Gly-Pro-Arg-Pro-AA of following formula,
AA is Ala residue, Glu residue, Gly residue and Pro residue in formula.
2. the preparation method of the warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-AA of claim 1, this method comprises:
1) warfarin -4-O- benzyl acetate is synthesized；
2) warfarin -4-O- acetic acid is synthesized；
3) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes Gly- using liquid phase method Pro-OBzl；
3) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes warfarin-using liquid phase method 4-O- acetyl-Gly-Pro；
4) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes Arg using liquid phase method (NO2)-Pro-Ala-OBzl,Arg(NO2)-Pro-Glu(OBzl)-OBzl,Arg(NO2)-Pro-Gly-OBzl,Arg(NO2)- Pro-Pro-OBzl；
5) using dicyclohexylcarbodiimide as condensing agent, I-hydroxybenzotriazole is catalyst, synthesizes warfarin-using liquid phase method 4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Ala-OBzl, warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro- Glu (OBzl)-OBzl, warfarin -4-O- acetyl-Gly-Pro-Arg (NO2)-Pro-Gly-OBzl, warfarin -4-O- acetyl - Gly-Pro-Arg(NO2)-Pro-Pro-OBzl；
6) hydrogenolysis Deprotection synthesizes warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Ala, warfarin -4-O- acetyl - Gly-Pro-Arg-Pro-Glu, warfarin -4-O- acetyl-Gly-Pro-Arg-Pro-Gly, warfarin -4-O- acetyl-Gly- Pro-Arg-Pro-Pro。
3. warfarin -4-O- acetyl-the Gly-Pro-Arg-Pro-AA of claim 1 is in preparing anti-phlebothrombosis drug Using.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113929736A (en) * 2020-06-29 2022-01-14 首都医科大学 Gly-Pro-Arg-Pro-oxyethyl aminocarbonyl warfarin, synthesis, activity and application thereof
CN113929742A (en) * 2020-06-29 2022-01-14 首都医科大学 warfarin-4-O-acetyl-Gly-Pro-Arg-Pro-AA 2 and synthesis, activity and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993023085A1 (en) * 1992-05-21 1993-11-25 Diatech, Inc. TECHNETIUM-99m LABELED PEPTIDES FOR THROMBUS IMAGING
WO2000009076A2 (en) * 1998-08-17 2000-02-24 Thomas Jefferson University Imaging with tc-99m labeled fibrin-alpha-chain peptide
CN1286262A (en) * 2000-09-12 2001-03-07 南京大学 Anticoagulant protein mediated by anticoagulant protein of human placenta
CN101267843A (en) * 2005-08-04 2008-09-17 哈莫斯塔蒂斯有限公司 Artificial platelets

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993023085A1 (en) * 1992-05-21 1993-11-25 Diatech, Inc. TECHNETIUM-99m LABELED PEPTIDES FOR THROMBUS IMAGING
WO2000009076A2 (en) * 1998-08-17 2000-02-24 Thomas Jefferson University Imaging with tc-99m labeled fibrin-alpha-chain peptide
CN1286262A (en) * 2000-09-12 2001-03-07 南京大学 Anticoagulant protein mediated by anticoagulant protein of human placenta
CN101267843A (en) * 2005-08-04 2008-09-17 哈莫斯塔蒂斯有限公司 Artificial platelets

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MARINA ZACCHIGNA等: "Improvement of warfarin biopharmaceutics by conjugation with poly(ethylene glycol)", 《EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES》 *
R. SUHAS等: "Synthesis of quinazolinone conjugated shorter analogues of Bactenecin7 as potent antimicrobials", 《PROTEIN & PEPTIDE LETTERS》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113929736A (en) * 2020-06-29 2022-01-14 首都医科大学 Gly-Pro-Arg-Pro-oxyethyl aminocarbonyl warfarin, synthesis, activity and application thereof
CN113929742A (en) * 2020-06-29 2022-01-14 首都医科大学 warfarin-4-O-acetyl-Gly-Pro-Arg-Pro-AA 2 and synthesis, activity and application thereof
CN113929736B (en) * 2020-06-29 2024-01-26 首都医科大学 Gly-Pro-Arg-Pro-oxaminocarbonyl warfarin, synthesis, activity and application thereof

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