CN106279363B - Pentamyloxycarbonylamino propionyl-GRPAK peptides, their preparation, activity and use - Google Patents
Pentamyloxycarbonylamino propionyl-GRPAK peptides, their preparation, activity and use Download PDFInfo
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Abstract
The invention discloses pentamethoxyl amido carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val with the following formula, discloses a preparation method thereof, discloses antithrombotic activity thereof, discloses thrombolytic activity thereof and discloses the effect of treating a rat suffering from apoplexy, thereby disclosing the application of the pentamethoxyl carbonyl propionyl-Lys in preparing antithrombotic drugs, thrombolytic drugs and drugs for treating ischemic stroke.
Description
Technical Field
The invention relates to pentamethoxyl tryptophanyl carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val, relates to a preparation method thereof, relates to antithrombotic activity thereof, relates to thrombolytic activity thereof and relates to an effect of treating ischemic stroke, so the invention relates to application thereof in preparing free radical scavenging medicaments, antithrombotic medicaments, thrombolytic medicaments and ischemic stroke medicaments. The invention belongs to the field of biological medicine.
Background
Ischemic stroke is a common and serious cerebrovascular disease, and is characterized by high morbidity, high fatality rate, high disability rate and high recurrence rate. At present, the clinical treatment of ischemic stroke faces the reality that no effective medicine exists, and especially, patients with stroke for more than 4 hours are not dead or are disabled. The invention of the medicine effective for patients with stroke for more than 4 hours is an important clinical requirement. The inventors have discovered that the imidazoline compound of formula I exhibits excellent therapeutic effects in a rat ischemic stroke model with stroke duration of 24 h. That is, the imidazoline compound of formula II is continuously injected intravenously for 6 days, and has excellent curative effect 1 time per day, the first dosage is 5 mu mol/kg, and the last 5 dosages are 2 mu mol/kg. In the formula aa1And aa2Can be present simultaneously aa1Exist but aa2Absent, or both; when aa1And aa2In the meantime, aa1Is R (Arg), and aa2G (Gly), A (Ala) or Q (Gln); when aa1Exist but aa2In absence aa1Is R (Arg); aa3It may be S (Ser), V (Val) or F (Phe). Since the 2-position of the imidazoline compound of formula II is 4-oxyacetyl-Lys. The side chain amino group and the main chain carboxyl group of the Lys are respectively connected with the RGD antithrombotic tetrapeptide and the ARPAK thrombolytic peptide, so the structure is more complicated and needs to be simplified.
Through 3 years of experimental research, the inventor finds that unexpected technical effects with simple structure and good curative effect can be obtained by replacing 2- (4-oxyacetyl) phenyl-4, 4,5, 5-tetramethyl-1, 3-dioxyimidazoline imidazolinyl of the formula I with pentamethoxyl amino carbonyl propionyl. In light of this finding, the inventors have devised the present invention.
Disclosure of Invention
One aspect of the present invention provides pentamethoxytetracycline carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val of the formula.
The invention also provides a preparation method of pentamethoxyl amido carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val, which comprises the following steps:
1) preparing pentamethoxyl tryptophane carbonyl propionic acid;
2) preparation of Boc-Arg (NO)2)-Gly-OBzl；
3) Preparation of Boc-Arg (NO)2)-Gly；
4) Preparing Boc-Asp (OBzl) -Val-OBzl;
5) preparing HCl & Asp (OBzl) -Val-OBzl;
6) preparation of Boc-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
7) Preparation of HCl. Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
8) Preparation of Fmoc-Lys (Boc) -Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
9) Preparation of Fmoc-Lys-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
10) Preparing Boc-Pro-Ala-OBzl;
11) preparing Boc-Pro-Ala;
12) preparation of Boc-Pro-Ala-Lys (Z) -OBzl;
13) preparation of HCl Pro-Ala-Lys (Z) -OBzl;
14) preparation of Boc-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
15) Preparation of HCl. Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
16) Preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
17) Preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)；
l8) preparation of Fmoc-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
19)Lys(Boc-Gly-Arg(NO2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
20) Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2) reacting-Gly-Asp (OBzl) -Val-OBzl with pentamethoxyl tryptophane carbonyl propionic acid to obtain pentamethoxyl tryptophane carbonyl propionyl-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
21) Pentamethoxytryptophanylcarbonylpropionyl-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2) Deprotection of-Gly-Asp (OBzl) -Val-OBzl to obtain pentamethoxyl tryptophane carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val.
The third content of the invention is to evaluate the antithrombotic activity, thrombolytic activity and ischemic stroke treatment effect of the pentamethoxyl tryptophane carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val.
Drawings
FIG. 1. Pentamethoxytryptophanylcarbonylpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val (1) synthetic route: (a) DCC, HOBt, NMM, THF; (b)2N NaOH, THF; (c)4N hydrogen chloride-ethyl acetate solution; (d) piperidine/dmf (e) succinic anhydride; (f) TFA/TFMSA.
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 general method for peptide ligation
Dissolving 1mmol carboxyl-terminal compound in dry THF, adding 1.2mmol N-hydroxy benzotriazole (HOBt) and 1.2mmol N, N-dicyclohexyl carbodiimide (DCC) dissolved in dry THF under stirring in ice bath, stirring for 0.5h, dissolving 1.05mmol amino-terminal compound in dry THF, adding into the above reaction solution, adjusting pH to 9 with N-methylmorpholine (NMM), stirring at room temperature for 6h, and TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 10/1) to obtain the target compound after separation and purification.
EXAMPLE 2 general procedure for removal of N-t-Butoxycarbonyl protecting group
Dissolving 1mmol of compound containing N-tert-butoxycarbonyl protecting group with small amount of dry ethyl acetate, adding 10mL of 4N hydrogen chloride/ethyl acetate solution under stirring in ice bath, and stirring in ice bathStirring for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. 5ml of anhydrous ethyl acetate was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
EXAMPLE 3 general method for hydrolytic removal of benzyl ester protecting group
Dissolving the compound containing benzyl ester protecting group in methanol, slowly adding 2M NaOH aqueous solution dropwise under ice bath and stirring, adjusting pH to 12, reacting for 5h, and TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. Slowly dropwise adding saturated KHSO under ice-bath stirring4Adjusting pH of the aqueous solution to 7, concentrating under reduced pressure to remove methanol, and slowly adding saturated KHSO dropwise into the remaining aqueous solution under stirring in ice bath4Adjusting pH of the aqueous solution to 3, extracting with ethyl acetate for 3 times, washing the combined ethyl acetate layers with saturated aqueous NaCl solution for 3 times, and adding anhydrous Na2SO4Drying, filtering, and concentrating the filtrate under reduced pressure to obtain the target compound.
EXAMPLE 4 general procedure for the hydrogenolysis removal of benzyl ester protecting group
Dissolving the compound containing benzyl ester protecting group in methanol, adding Pd/C (20% of the reactant), extracting air from the reaction system under reduced pressure, introducing hydrogen, stirring at room temperature for 10h, and TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. And filtering to remove Pd/C, and concentrating the filtrate under reduced pressure to obtain the target compound.
20) Preparing pentamethoxyl tryptophane carbonyl propionic acid;
EXAMPLE 5 preparation of Pentamethoxytryptophane Carbonylpropionic acid
Dissolving 1.9g (10.0mmol) of pentamethoxytetramine in dry THF, adding 1.20g (12.0mmol) of succinic anhydride while stirring in ice bath, adjusting pH to 9 with NMM, stirring at room temperature for 6h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, and sequentially adding 5% KHSO4The aqueous solution was washed 3 times, and the saturated NaCl aqueous solution was washed 3 times. The combined ethyl acetate layer was washed with waterWater Na2SO4Drying, filtration and concentration of the filtrate under reduced pressure to dryness gave 2.81g (96.9%) of the title compound as a pale yellow solid. ESI-MS (m/e): 289[ M-H ]]-。
EXAMPLE 6 preparation of Boc-Pro-Ala-OBzl
The peptide grafting method of example 1 was followed to dissolve 0.84g (3.90mmol) of Boc-Pro carboxy-terminal compound in dry THF, add 0.63g (4.68mmol) of HOBt and 0.96g (4.68mmol) of DCC dissolved in dry THF while stirring in ice bath, stir for 0.5h, dissolve 1.30g (3.7mmol) of HCl. Ala-OBzl amino-terminal compound in dry THF, add to the above reaction solution, adjust NMM to pH9, stir at room temperature for 6h, TLC (petroleum ether/acetone, 3/2) showed complete disappearance of carboxy-terminal material and reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 100/1) to yield 1g (72%) of the title compound as a colorless solid. ESI-MS (m/e): 377[ M + H]+。
EXAMPLE 7 preparation of Boc-Pro-Ala
Following the general procedure for hydrolytic deprotection of benzyl ester in example 3, 1g (2.66mmol) of Boc-Pro-Ala-OBzl was dissolved in methanol, adjusted to pH12 by slowly adding dropwise 2M aqueous NaOH solution with stirring in ice bath, reacted for 5h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. Slowly dropwise adding saturated KHSO under ice-bath stirring4Adjusting pH of the aqueous solution to 7, concentrating under reduced pressure to remove methanol, and slowly adding saturated KHSO dropwise into the remaining aqueous solution under stirring in ice bath4Adjusting pH of the aqueous solution to 3, extracting with ethyl acetate for 3 times, washing the combined ethyl acetate layer with saturated aqueous NaCl solution for 3 times, and adding anhydrous Na2SO4Drying, filtration and concentration of the filtrate under reduced pressure gave 0.7g (92%) of the title compound as a colorless solid. ESI-MS (m/e): 285[ M-H ]]-。
EXAMPLE 8 preparation of Boc-Pro-Ala-Lys (Z) -OBzl
The peptide grafting method of example 1 was followed to dissolve 1.0g (3.7mmol) of Boc-Pro-Ala carboxy-terminal compound in dry THF, add 0.60g (4.44mmol) of HOBt and 0.91g (4.44mmol) of DCC dissolved in dry THF in sequence under ice-bath stirring, stir for 0.5h, dissolve 1.25g (3.10mmol) of HCl. Lys- (Z) -OBzl amino-terminal compound in dry THF, add to the above reaction solution, adjust NMM to pH9, stir at room temperature for 6h, TLC (petroleum ether/acetone, 1/1) showed complete disappearance of carboxy-terminal material and reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 100/1) to yield 1.5g (76%) of the title compound as a colorless solid. ESI-MS (m/e): 639[ M + H]+。
EXAMPLE 9 preparation of HCl Pro-Ala-Lys (Z) -OBzl
1mmol 1g (1.6mmol) of Boc-Pro-Ala-Lys (Z) -OBzl was dissolved with a small amount of dry ethyl acetate according to the general procedure for N-t-butyloxycarbonyl removal in example 2, 10mL of 4N hydrogen chloride/ethyl acetate solution was added with stirring in ice bath, stirring in ice bath was carried out for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. 5ml of anhydrous ethyl acetate was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
EXAMPLE 10 preparation of Boc-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl
Peptide grafting according to example 1 from 1.5g (4.60mmol) Boc-Arg (NO)2) The carboxy-terminal compound was dissolved in dry THF, 0.75g (5.5mmol) of HOBt and 1.14g (5.5mmol) of DCC dissolved in dry THF were added sequentially with stirring in an ice bath, stirred for 0.5h, and 1.30g (3.7mmol) of HCl. Ala-OBzl amino-terminal compound was dissolved in dry THFTHF was dried and added to the reaction solution, NMM adjusted to pH9, stirred at room temperature for 6h, TLC (CH)2Cl2∶CH3OH,20/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH,20/1) to yield 4.46g (69%) of the title compound as a colorless solid. ESI-MS (m/e): 841[ M + H]+。
EXAMPLE 11 preparation of HCl Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl
Following example 2 general procedure for N-t-Butoxycarbonyl removal of the protecting group 1g (1.19mmol) of Boc-Arg, (NO)2) -Pro-Ala-Lys (Z) -OBzl is dissolved in a small amount of dry ethyl acetate, 10mL of 4N hydrogen chloride/ethyl acetate solution is added with stirring in ice bath, and the mixture is stirred in ice bath for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. 5ml of anhydrous ethyl acetate was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
EXAMPLE 12 preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl
The peptide grafting method of example 1 was followed to dissolve 2.58g (14.75mmol) of Boc-Gly carboxy-terminal compound in dry THF, add 2.39g (17.7mmol) of HOBt and 3.65g (17.7mmol) of DCC dissolved in dry THF while stirring in ice bath, stir for 0.5h, and add 10.90g (14.05mmol) of HCl Arg (NO)2) Dissolving the amino-terminal compound (Pro-Ala-Lys) (Z) -OBzl in dry THF, adding into the above reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, TLC (CH)2Cl2∶CH3OH, 15/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, and decompressing the filtrateConcentrating, dissolving with ethyl acetate, sequentially adding saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 15/1) to yield 6.00g (48%) of the title compound as a colorless solid. ESI-MS (m/e): 897[ M + H]+。
EXAMPLE 13 preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)
Following the general procedure for hydrolytic deprotection of benzyl ester in example 3, 1g (1.11mmol) of Boc-Gly-Arg (NO)2) -Pro-Ala-Lys (Z) -OBzl in methanol, slowly adding 2M NaOH aqueous solution dropwise with ice bath and stirring, adjusting to pH12, reacting for 5h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. Slowly dropwise adding saturated KHSO under ice-bath stirring4Adjusting pH of the aqueous solution to 7, concentrating under reduced pressure to remove methanol, and slowly adding saturated KHSO dropwise into the remaining aqueous solution under stirring in ice bath4Adjusting pH of the aqueous solution to 3, extracting with ethyl acetate for 3 times, washing the combined ethyl acetate layer with saturated aqueous NaCl solution for 3 times, and adding anhydrous Na2SO4Drying, filtration and concentration of the filtrate under reduced pressure gave 0.86g (97%) of the title compound as a colorless solid. ESI-MS (m/e): 806[ M-H]-。
EXAMPLE 14 preparation of Boc-Arg (NO)2)-Gly-OBzl
The peptide grafting procedure was as in example 1 from 4.98g (15.5mmol) of Boc-Arg (NO)2) Dissolving the carboxyl terminal compound in dry THF, adding 2.51g (18.6mmol) of HOBt and 3.81g (18.6mmol) of DCC dissolved in dry THF under stirring in ice bath, stirring for 0.5h, dissolving 6.49g (15.0mmol) of HCl-Gly-OBzl amino terminal compound in dry THF, adding into the above reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, TLC (CH)2Cl2∶CH3OH, 15/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Dissolving in waterWashing with 5% KHSO 3 times, 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 60/1) to yield 6g (87%) of the title compound as a colorless solid. ESI-MS (m/e): 467[ M + H ]]+。
EXAMPLE 15 preparation of Boc-Arg (NO)2)-Gly
Following the general procedure for hydrolytic deprotection of benzyl ester in example 3, 1g (2.14mmol) of Boc-Arg (NO)2) dissolving-Gly-OBzl in methanol, slowly adding 2M NaOH aqueous solution dropwise under ice bath and stirring, adjusting pH to 12, reacting for 5h, and TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. Slowly dropwise adding saturated KHSO under ice-bath stirring4Adjusting pH of the aqueous solution to 7, concentrating under reduced pressure to remove methanol, and slowly adding saturated KHSO dropwise into the remaining aqueous solution under stirring in ice bath4Adjusting pH of the aqueous solution to 3, extracting with ethyl acetate for 3 times, washing the combined ethyl acetate layer with saturated aqueous NaCl solution for 3 times, and adding anhydrous Na2SO4Drying, filtration and concentration of the filtrate under reduced pressure gave 0.797g (99%) of the title compound as a colorless solid. ESI-MS (m/e): 376[ M-H]-。
EXAMPLE 16 preparation of Boc-Asp (OBzl) -Val-OBzl
The peptide grafting procedure of example 1 was followed by dissolving 4.5g (13.85mmol) Boc-Asp (OBzl) carboxy-terminal compound in dry THF, adding 2.24g (16.62mmol) HOBt and 3.42g (16.62mmol) DCC dissolved in dry THF in sequence with stirring in an ice bath, stirring for 0.5h, dissolving 5.0g (13.2mmol) HCl Val-OBzl amino-terminal compound in dry THF, adding to the reaction mixture NMM to pH9, stirring at room temperature for 6h, TLC (CH)2Cl2∶CH3OH, 30/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO3Dissolving in waterWashing with liquid 3 times and with saturated aqueous NaCl solution 3 times. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 60/1) to yield 6g (91%) of the title compound as a colorless solid. ESI-MS (m/e): 512[ M + H]+。
EXAMPLE 17 preparation of HCl Asp (OBzl) -Val-OBzl
Following example 2 general procedure for N-t-butyloxycarbonyl deprotection 1g (1.95mmol) of Boc-Asp (OBzl) -Val-OBzl was dissolved with a small amount of dry ethyl acetate, 10mL of 4N hydrogen chloride/ethyl acetate solution was added with stirring in ice bath, stirring in ice bath for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. 5ml of anhydrous ethyl acetate was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
EXAMPLE 18 preparation of Boc-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl
The peptide grafting procedure was as in example 1 from 5.03g (15.5mmol) of Boc-Arg (NO)2) Dissolving the-Gly carboxyl terminal compound in dry THF, adding 2.51g (18.6mmol) of HOBt and 3.81g (18.6mmol) of DCC dissolved in dry THF under stirring in ice bath, stirring for 0.5h, dissolving 6g (13.37mmol) of HCl & Asp (OBzl) -Val-OBzl amino terminal compound in dry THF, adding into the reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, and TLC (CH)2Cl2∶CH3OH,20/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and saturation of NaThe aqueous Cl solution was washed 3 times. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH,20/1) to yield 7.02g (68%) of the title compound as a colorless solid。ESI-MS(m/e)：771[M+H]+。
EXAMPLE 19 preparation of HCl Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl
Following example 2 general procedure for deprotection of N-t-butyloxycarbonyl 1g (1.30mmol) of Boc-Arg (NO)2) -Gly-Asp (OBzl) -Val-OBzl was dissolved in small amount of dry ethyl acetate, 10mL of 4N HCl/ethyl acetate solution was added with stirring in ice bath, and the mixture was stirred in ice bath for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. 5ml of anhydrous ethyl acetate was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
EXAMPLE 20 preparation of Fmoc-Lys (Boc) -Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl
The peptide grafting procedure of example 1 was followed to dissolve 7.82g (16.68mmol) of Fmoc-Lys (Boc) carboxy-terminal compound in dry THF, add 2.71g (20.0mmol) of HOBt and 4.12g (20.0mmol) of DCC dissolved in dry THF while stirring in ice bath, stir for 0.5h, and add 11.796g (16.68mmol) of HCl-Arg (NO)2) Dissolving the amino-terminal compound of (E) -Gly-Asp (OBzl) -Val-OBzl in dry THF, adding into the above reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, TLC (CH)2Cl2∶CH3OH,20/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 15/1) to yield 12.7g (68%) of the title compound as a colorless solid. ESI-MS (m/e): 1121[ M + H ]]+。
EXAMPLE 21 preparation of Fmoc-Lys-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl
Pressing to realEXAMPLE 2 deprotection of N-tert-Butoxycarbonyl protecting group 1g (0.89mmol) of Fmoc-Lys (Boc) -Arg (NO)2) -Gly-Asp- (OBzl) -Val-OBzl was dissolved in small amount of dry ethyl acetate, 10mL of 4N HCl/ethyl acetate solution was added with stirring in ice bath, and the mixture was stirred in ice bath for 1-2h, TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of starting material and the reaction was complete. The reaction solution was concentrated under reduced pressure. The residue was added with 5ml of anhydrous ethyl acetate, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. 5ml of anhydrous ether was added to the residue, and the solution was concentrated to dryness under reduced pressure. This operation was repeated 3 times. The obtained target compound was directly used in the next reaction.
Example 22 preparation
Fmoc-Lys(Boc-Gly-Arg(NO2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl
Peptide grafting according to example 1 from 2.37g (2.93mmol) Boc-Gly-Arg (NO)2) Pro-Ala-Lys (Z) carboxy-terminal Compound dissolved in dry THF, 0.47g (3.51mmol) of HOBt and 0.72g (3.51mmol) of DCC dissolved in dry THF were added sequentially with stirring in an ice bath, stirred for 0.5h, and 3.09g (2.93mmol) of Fmoc-Lys-Arg (NO)2) Dissolving the amino-terminal compound (Gly-Asp) (OBzl) -Val-OBzl in dry THF, adding into the above reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, and performing TLC (CHCl)3/CH3OH, 10/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH, 10/1) to yield 1.70g (48%) of the title compound as a colorless solid. ESI-MS (m/e): 1826[ M + H]+。
Example 23 preparation
Lys(Boc-Gly-Arg(NO2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl
0.5g (0) was added under ice bath.27mmol)Fmoc-Lys(Boc-Gly-Arg(NO2)-Pro-Ala-Lys(Z))-Arg(NO2) -Gly-Asp (OBzl) -Val-OBzl dissolved in 1mL 20% piperidine/DMF (N, N-dimethylformamide) for 30min, TLC (CHCl)3/CH3OH,5/1) showed the disappearance of the starting point. Ether was added to precipitate a solid which was filtered to give the title compound as a colorless solid which was used directly in the next reaction.
EXAMPLE 24 preparation of pentamethyloxycarpropionyl-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl
The peptide grafting method as in example 1 was followed by dissolving 20.7mg (0.069mmol) of the carboxy-terminal Pentamyloxychromanocarboxypropionic acid compound in dry THF, adding 0.012g (0.083mmol) of HOBt and 0.017g (0.083mmol) of DCC dissolved in dry THF in that order while stirring in an ice bath, stirring for 0.5h, and adding 0.1g (0.062mmol) of Lys (Boc-Pro-Ala-Lys- (Z)) -Arg (NO)2) Dissolving the amino-terminal compound (Gly-Asp) (OBzl) -Val-OBzl in dry THF, adding into the above reaction solution, adjusting NMM to pH9, stirring at room temperature for 6h, and performing TLC (CHCl)3∶CH3OH,8/1) showed complete disappearance of the carboxy-terminal starting material and the reaction was complete. Filtering to remove DCU, concentrating the filtrate under reduced pressure, dissolving with ethyl acetate, sequentially dissolving the obtained solution with saturated NaHCO3Washing with 5% KHSO 3 times and 3 times with saturated NaCl aqueous solution4Washing with aqueous solution 3 times, washing with saturated aqueous NaCl solution 3 times, 5% NaHCO33 washes with aqueous solution and 3 washes with saturated aqueous NaCl solution. The combined ethyl acetate layers were washed with anhydrous Na2SO4Drying, filtering, concentrating the filtrate under reduced pressure to dryness, and separating the column layer (CHCl)3/CH3OH,8/1) to yield 0.09g (48%) of the title compound. ESI-MS (m/e): 1861[ M + H]+。
EXAMPLE 25 preparation of pentamethyloxycarbonylpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val (1)
100mg (0.055mmol) pentamethyloxytryptophanylcarbonylpropionyl-Lys (Boc-Gly-Arg (NO) under ice bath2)-Pro-Ala-Lys(Z))-Arg(NO2) -Gly-Asp (OBzl) -Val-OBzl is mixed with 1mL of trifluoroacetic acid and 0.33mL of trifluoromethanesulfonic acid, stirred for 40min, poured into 50mL of anhydrous ether and precipitated. Washing the solid with anhydrous diethyl ether repeatedly and concentrating under reduced pressure to obtain light yellowThe powder was dissolved in water, adjusted to pH8 with ammonia, desalted via Sephadex G10, purified using a C18 column (eluent methanol water 5: 95), and the collected fractions were lyophilized to give 30mg (29%) of the title compound as a colorless solid. Mp 182-;
ESI-MS(m/e)：1355[M+H]+；1H-NMR(300MHz，DMSO-d6)：8/ppm＝10.638(s，1H)，9.5-7.5(m，4H)，7.222(d，J＝8.8Hz，1H)，7.098(s，1H)，7.012(d，J＝1.6Hz，1H)，6.711(d，J＝8.8Hz，1H)，4.6-4.0(m，9H)，3.759(s，3H)，3.7-3.5(m，5H)，3.5-3.0(m，15H)，3.0-2.5(m，4H)，2.5-2.0(m，8H)，2.0-1.5(m，16H)，1.5-1.0(m，12H)，0.754(d，J＝7.2Hz，6H)。
test example 1 evaluation of antithrombotic Activity of Compound 1
Male SD rats (200. + -.20 g) were randomly divided into groups of 10 animals each, kept for 1 day and stopped overnight. After 30min of gavage administration of a physiological saline solution of Compound 1 (dose: 100nmol/kg) or aspirin (dose: 167. mu. mol/kg) or a physiological saline (dose: 10ml/kg), rats were anesthetized with a physiological saline solution of 20% Ulipraz, followed by surgery. The right carotid artery and the left jugular vein of the rat were isolated, accurately weighed silk was placed in the bypass cannula, one end of the tube was inserted into the left vein and the other end was inserted into the right artery and injected with 0.2mL heparin sodium anticoagulation. Allowing blood flow to flow from the right artery through the bypass cannula into the left vein, taking out the thread with thrombus after 15min, weighing, calculating the weight of the thread before and after blood circulation, and performing t-test to obtain the weight of the thrombus represented by the average value + -SD mg and representing the antithrombotic activity. The data are shown in Table 1. The results show that oral administration of 100nmol/kg of Compound 1 is effective in inhibiting thrombosis. After the structure is simplified, the technical effect is obvious.
TABLE 1 antithrombotic Activity of Compound 1
n is 10; a) the ratio of p to the physiological saline is less than 0.01.
Experimental example 2 evaluation of thrombolytic Activity of Compound 1
SD rats (male, 200 + -20 g) according to 1200mg/kg-1The dosage of the preparation is to inject the physiological saline solution of urethane into the abdominal cavity for anesthesia. Fixing the rat in a supine position after anesthesia, separating the right common carotid artery of the rat, clamping an artery clamp at the proximal end, respectively penetrating the proximal end and the distal end into an operation line, ligating the operation line at the distal end, inserting a tube at the distal end, loosening the artery clamp, taking out about 1mL of arterial blood, and placing the arterial blood in a 1mL centrifuge tube. 0.1ml of rat arterial blood is injected into a vertically fixed rubber tube (15 mm in length, 2.5mm in inner diameter, 5.0mm in outer diameter, the tube bottom is sealed by a rubber plug, and the para membrane is tightly sealed), and then a fixing bolt of a stainless steel thrombus is rapidly inserted into the tube (the thrombus fixing spiral is wound by a stainless steel wire with the diameter of 0.2mm, the spiral part is 10mm long and internally contains 15 spiral rings, the diameter of the spiral rings is 1.0mm, and the support handle is connected with the spiral and is about 7.0mm long and in a question mark shape). After blood coagulation for 45min, the thrombus-immobilized spiral wrapped by the thrombus was carefully removed from the glass tube and weighed accurately.
The bypass cannula consists of three parts, wherein the middle section is a polyethylene rubber tube with the length of 60.0mm and the inner diameter of 3.5 mm; both ends are 100.0mm long, and internal diameter 1.0mm, the same polyethylene pipe of external diameter 2.0mm, and this pipe one end is drawn into the sharp pipe, and is about 10.0mm long (being used for inserting rat carotid artery and vein), and the external diameter is 1.0mm, and the outside cover section of its other end is long for 7.0mm, and the external diameter is 3.5 mm's polyethylene pipe (in being used for inserting the polyethylene rubber tube in middle section), and the inner wall of 3 sections pipes all needs silanization (1% silicon oil ether solution). The thrombus-wrapped thrombus fixing spiral is placed in the middle section polyethylene rubber tube, and the other two ends of the rubber tube are respectively sleeved with the thickened ends of the two polyethylenes, so that blood leakage can be avoided in the circulating process. The tube was filled with heparin normal saline solution (50IU/kg) through the tip end with a syringe to remove air bubbles for use.
The left external jugular vein of separation rat, proximal end and distal end penetrate the operation line respectively, and the blood vessel of ligature distal end cuts a osculum on the left external jugular vein that exposes, inserts the bypass pipeline taper pipe that has been prepared above-mentioned into left external jugular vein opening part by the osculum, keeps away from bypass pipe middle section (contains the thrombus fixed spiral of accurate weighing) internal thrombus fixed spiral simultaneously. An accurate amount of a physiological saline solution (50IU/kg) of heparin sodium was injected through the tip tube at the other end with a syringe, at which time the syringe was not removed from the polyethylene tube, and the flexible tube between the syringe and the polyethylene tube was clamped with an artery clamp. Stopping bleeding by an artery clamp at the proximal end of the right common carotid artery, ligating the distal end, cutting a small opening of the right common carotid artery at a position short of the artery clamp, pulling out the injector from the tip of the polyethylene tube, and inserting the tip of the polyethylene tube into the proximal end of the oblique opening of the artery. Both ends of the bypass pipeline are used for fixing the artery and the vein by using a No. 4 surgical suture.
Physiological saline (3ml/kg) or a physiological saline solution of urokinase (dosage of 20000IU/kg) or a physiological saline solution of compound 1 (dosage of 100nmol/kg) was passed through the middle section of the bypass tube (containing the thrombus fixing helix accurately weighed) with a scalp needle, inserted into the proximal venous end remote from the thrombus fixing helix, and the arterial clamp was released, allowing blood flow from the artery to the vein through the bypass tube. The solution in the syringe is slowly injected into the blood, and acts on the spiral thrombus through the blood circulation in the venous-cardiac-arterial order. After 1h of blood circulation, the thrombus-anchoring helix was removed from the bypass tube and accurately weighed. The weight difference of the thrombus before and after spiral blood circulation of the immobilized thrombus in the bypass channel of each rat was calculated, expressed as mean value ± SD mg and representing the thrombolytic activity, and subjected to t-test. The data are shown in Table 2. The results show that 100nmol/kg of Compound 1 effectively dissolves the thrombus formed. The thrombolytic activity of 100nmol/kg of compound 1 is comparable to 20000IU/kg urokinase. After the structure is simplified, the technical effect is obvious.
TABLE 2 thrombolytic Activity of Compound 1
n is 10; a) the ratio p to normal saline is less than 0.01, and the ratio p to urokinase is more than 0.05.
Experimental example 3 evaluation of therapeutic Effect of Compound 1 on ischemic apoplexy rats
A2 cm long incision was made vertically in the middle of the neck of male SD rats (body weight 300. + -.20 g), and the right common carotid artery, external carotid artery and internal carotid artery were isolated along the intramuscular side edge of the sternocleidomastoid muscle. Respectively clamping an opening of an internal carotid artery and a proximal end of a common carotid artery by using a noninvasive artery clamp, ligating a distal end of an external carotid artery, cutting a small opening on the external carotid artery, loosening the artery clamp at the proximal end of the common carotid artery, taking 10 mu l of blood, and then clamping the proximal end of the common carotid artery by using the noninvasive artery clamp. The obtained 10. mu.l of blood was placed in a 1ml EP tube at normal temperature for 30 minutes to coagulate the blood, and then transferred to a-20 ℃ refrigerator and left for 1 hour to make the blood clot firm. Rats were anesthetized with 10% chloral hydrate by intraperitoneal injection at a dose of 400 mg/kg. The blood clot was removed, 1ml of physiological saline was added, the blood clot was crushed with a steel spatula into fine thrombus pieces of uniform size, a suspension of the fine thrombus was prepared and transferred to a 1ml syringe. Loosening the artery clamp at the proximal end of the common carotid artery, slowly injecting 1ml of thrombus suspension into the brain of a rat from the external carotid artery of the rat to the proximal end through the internal carotid artery, then ligating the proximal end of the external carotid artery, and opening the internal carotid artery and the common carotid artery to obtain the artery clamp to restore blood flow. Waiting for wakeup. The degree of neurological deficit was assessed by the Zealonga method 24 hours after the rats were awakened. Score 0 indicates no sign of neurological deficit, score 1 indicates that the intact forelimb cannot stretch, score 2 indicates walking to the intact side, score 3 indicates turning to the intact side and walking in a tail-end-collision manner, score 4 indicates that the disorder is not self-walking, and score 5 indicates death. And grouping according to the score average. Each group of rats was injected with compound 1 via the tail vein 1 time per day at a dose of 100 nmol/kg. Injections were given continuously for 6 days, and scored daily. The results are shown in Table 3. The data indicate that compound 1 treatment for 6 consecutive days resulted in a neurobiological score of 1 in all rats with 8 cerebral ischemia for 24 hours. Since the first dose of compound required 5. mu. mol/kg and the last 5 maintenance doses 2. mu. mol/kg, unlike the already disclosed compounds, the 6 doses of compound 1 were all 100 nmol/kg. Thus, the first and maintenance doses were reduced by 50 and 20 times, respectively. The invention obtains unexpected technical effect by adding the advantage of simplified structure.
TABLE 3 Effect of Compound 1 on neurobiological scores in rats with 24h cerebral ischemia on 6 consecutive days
n＝8。
Claims (5)
1. Pentamethoxytryptophanylcarbonylpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val of formula:
2. a process for the preparation of pentamethyloxycarbonylpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val according to claim 1, which comprises the steps of:
1) preparing pentamethoxyl tryptophane carbonyl propionic acid;
2) preparation of Boc-Arg (NO)2)-Gly-OBzl；
3) Preparation of Boc-Arg (NO)2)-Gly；
4) Preparing Boc-Asp (OBzl) -Val-OBzl;
5) preparing HCl & Asp (OBzl) -Val-OBzl;
6) preparation of Boc-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
7) Preparation of HCl. Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
8) Preparation of Fmoc-Lys (Boc) -Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
9) Preparation of Fmoc-Lys-Arg (NO)2)-Gly-Asp(OBzl)-Val-OBzl；
10) Preparing Boc-Pro-Ala-OBzl;
11) preparing Boc-Pro-Ala;
12) preparation of Boc-Pro-Ala-Lys (Z) -OBzl;
13) preparation of HCl Pro-Ala-Lys (Z) -OBzl;
14) preparation of Boc-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
15) Preparation of HCl. Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
16) Preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)-OBzl；
17) Preparation of Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z)；
18) Preparation of Fmoc-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
19) Preparation of Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
20) Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2) reacting-Gly-Asp (OBzl) -Val-OBzl with pentamethoxyl tryptophane carbonyl propionic acid to obtain pentamethoxyl tryptophane carbonyl propionyl-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2)-Gly-Asp(OBzl)-Val-OBzl；
21) Pentamethoxytryptophanylcarbonylpropionyl-Lys (Boc-Gly-Arg (NO)2)-Pro-Ala-Lys(Z))-Arg(NO2) Deprotection of-Gly-Asp (OBzl) -Val-OBzl to obtain pentamethoxyl tryptophane carbonyl propionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val.
3. Use of pentamethyloxycarpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val of claim 1 for the preparation of an antithrombotic agent.
4. Use of pentamethyloxycarpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val of claim 1 for the preparation of a thrombolytic drug.
5. Use of pentamethyloxycarbonylpropionyl-Lys (Gly-Arg-Pro-Ala-Lys) -Arg-Gly-Asp-Val according to claim 1 for the preparation of a medicament for the treatment of ischemic stroke.
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| CN103159835A (en) * | 2011-12-13 | 2013-06-19 | 首都医科大学 | Lys and oligopeptide-modified curcumin derivatives, and synthesis and medical application thereof |
| CN103665107A (en) * | 2012-09-05 | 2014-03-26 | 永光制药有限公司 | Novel compound combining functions of dissolving thrombus, scavenging free radicals and targeting thrombus, as well as preparation method and application thereof |
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| CN1356338A (en) * | 2001-10-25 | 2002-07-03 | 北京明心源科技有限公司 | Separation, synthesis and application in medicine for P6A'S metabolic product |
| CN103159835A (en) * | 2011-12-13 | 2013-06-19 | 首都医科大学 | Lys and oligopeptide-modified curcumin derivatives, and synthesis and medical application thereof |
| CN103665107A (en) * | 2012-09-05 | 2014-03-26 | 永光制药有限公司 | Novel compound combining functions of dissolving thrombus, scavenging free radicals and targeting thrombus, as well as preparation method and application thereof |
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