CN108610394A - A kind of quasi- peptides prepare purification process and application - Google Patents
A kind of quasi- peptides prepare purification process and application Download PDFInfo
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- CN108610394A CN108610394A CN201810426114.0A CN201810426114A CN108610394A CN 108610394 A CN108610394 A CN 108610394A CN 201810426114 A CN201810426114 A CN 201810426114A CN 108610394 A CN108610394 A CN 108610394A
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- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 1
- 230000003480 fibrinolytic effect Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000007574 infarction Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 108010044426 integrins Proteins 0.000 description 1
- 102000006495 integrins Human genes 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- IRAXRQFCCSHQDX-WBVHZDCISA-N methyl (2s)-2-(butoxycarbonylamino)-3-[[2-[(5r)-3-(4-carbamimidoylphenyl)-4,5-dihydro-1,2-oxazol-5-yl]acetyl]amino]propanoate Chemical compound O1[C@@H](CC(=O)NC[C@H](NC(=O)OCCCC)C(=O)OC)CC(C=2C=CC(=CC=2)C(N)=N)=N1 IRAXRQFCCSHQDX-WBVHZDCISA-N 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000010016 myocardial function Effects 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 230000036581 peripheral resistance Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 230000010118 platelet activation Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- 108091006082 receptor inhibitors Proteins 0.000 description 1
- 229950002267 roxifiban Drugs 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000036262 stenosis Effects 0.000 description 1
- 208000037804 stenosis Diseases 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- DSNBHJFQCNUKMA-SCKDECHMSA-N thromboxane A2 Chemical compound OC(=O)CCC\C=C/C[C@@H]1[C@@H](/C=C/[C@@H](O)CCCCC)O[C@@H]2O[C@H]1C2 DSNBHJFQCNUKMA-SCKDECHMSA-N 0.000 description 1
- 229960003425 tirofiban Drugs 0.000 description 1
- COKMIXFXJJXBQG-NRFANRHFSA-N tirofiban Chemical compound C1=CC(C[C@H](NS(=O)(=O)CCCC)C(O)=O)=CC=C1OCCCCC1CCNCC1 COKMIXFXJJXBQG-NRFANRHFSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 208000004043 venous thromboembolism Diseases 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0815—Tripeptides with the first amino acid being basic
- C07K5/0817—Tripeptides with the first amino acid being basic the first amino acid being Arg
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1019—Tetrapeptides with the first amino acid being basic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Chemical & Material Sciences (AREA)
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- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
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- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Hematology (AREA)
- Diabetes (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses quasi- peptide medicaments of a kind of anti-platelet aggregation and its preparation method and application.This compound is blood platelet GPIIb/IIIa receptor antagonists, with active Chinese drug component substance monomer lamp-dish flower acetic, it is modified for the RGD class peptide structures of II b/ of GP, III a receptors, II b/ of GP, the III a receptor-specifics of RGD class peptides not only can be improved, can also improve the solubility of lamp-dish flower acetic.The compound has the pharmacological action that antiplatelet gathers, it can be clinically used for preventing from dying suddenly caused by myocardial oxygen delivery arterial occlusion, heart attack, unstable angina, non-q wave myocardial infarction, coronary intervention, or can be used for patients with acute coronary syndrome.
Description
Technical Field
The invention belongs to the field of medicines, and relates to a platelet aggregation resistant peptide mimetic compound, which comprises a synthesis technology and a screening method thereof. The compound can be clinically used for preventing myocardial oxygen supply artery occlusion, heart attack, unstable angina, Q-wave-free myocardial infarction and sudden death caused by coronary intervention treatment, or can be used for patients with acute coronary syndrome.
Background
Thrombotic diseases are common cardiovascular and cerebrovascular diseases, and are various diseases caused by blood vessel stenosis and occlusion caused by thrombus, so that main organs are subjected to ischemia and infarction to cause dysfunction, the diseases are usually manifested as myocardial infarction, ischemic cerebral infarction and venous thromboembolism, the thrombotic diseases can affect various organs and systems of the whole body, and the morbidity, the disability rate and the fatality rate are high.
In recent years, great progress has been made in the study of platelet molecules and cell biology, and it is gradually recognized that fibrinogen and platelet GPIIb/IIIa receptor binding is the ultimate common pathway for platelet aggregation. The combination of the GPIIb/IIIa receptor and fibrinogen is the final common way in the process of inducing platelet aggregation by various platelet activators such as ADP, thrombin, collagen, epinephrine, arachidonic acid, TXA2 and the like, inhibits the pathway to prevent platelet aggregation, can prevent and treat thrombosis, and is expected to overcome the defects in the treatment of directly induced platelet activation by aspirin and clopidogrel to collagen, thrombin and the like.
The GPIIb/IIIa receptor is the most abundant integrin on the surface of platelet membrane and can recognize the peptide sequence RGD of adhesion protein molecule. Researches show that both the fibrinogen and the vWF of the adhesion molecules contain RGD sequences which are binding sites of the adhesion molecules and the GP IIb/IIIa receptors. Therefore, the GP IIb/IIIa receptor inhibitor synthesized based on the RGD sequence is used as a novel anti-platelet aggregation medicament to block the final path of platelet aggregation, and becomes a hot medicament for clinical research at home and abroad.
Currently, clinically developed or marketed inhibitors of the gpiib/iiia receptor, such as eptifibatide (FDA approved), tirofiban (FDA approved), abciximab (FDA approved), roxifiban, sirafiban, etc., especially the first few, have achieved certain clinical effects, but their bleeding side effects have limited their use.
The erigeron breviscapus is separated to obtain the active ingredient breviscapinum: the breviscapine and scutellarin have high medicinal value, are widely used in clinic, and can be prepared into injections, tablets, dripping pills and the like, and are mainly used for treating cardiovascular diseases. The erigeron breviscapus injection is a traditional Chinese medicine injection extracted from erigeron breviscapus and mainly contains flavonoid and phenolic acid, wherein the flavonoid is represented by scutellarin and is known as the main effective component of the erigeron breviscapus injection. In recent years, China has more reports about the application of erigeron breviscapus injection to treat Unstable Angina (UAP), and the curative effect shows better expectation, the main component of the erigeron breviscapus injection, namely total flavone, can reduce peripheral resistance, expand arteriole, improve the cardiovascular oxygen supply capability, improve the blood supply and the myocardial function, relieve the damage of myocardial cells in an anoxic period, increase the blood perfusion amount of tissues, improve microcirculation and metabolism and improve the macrophage immunologic function of organisms; simultaneously, the medicine can resist ischemia and anoxia caused by posterior pituitary and platelet aggregation caused by adenosine triphosphate, and has strong functions of inhibiting intravascular coagulation and promoting fibrinolytic activity. However, in clinical use, the erigeron breviscapus injection also has rare side effects of causing multiple organ function damage, anaphylactic shock, anaphylactic asthma, abnormal liver function, joint swelling and the like. Although the injection of scutellarin has good clinical effect on cardiovascular diseases, the injection is inconvenient to clinically administer and has rare side effects. The scutellarin oral preparation has the medicinal generation characteristics of extremely low bioavailability, wide metabolism, quick elimination and the like after being orally taken.
The inventor surprisingly discovers that the traditional Chinese herbal medicine scutellarin is used as an amino acid template, the RGD peptide structure of a GP IIb/IIIa receptor is modified, the GP IIb/IIIa receptor specificity of the RGD peptide is improved, the solubility of scutellarin is improved, and the platelet aggregation resistant peptide mimetic compound is developed and clinically used for preventing myocardial oxygen supply artery occlusion, heart attack, unstable angina, Q-wave-free myocardial infarction and sudden death caused by coronary intervention treatment or used for patients with acute coronary syndrome.
Disclosure of Invention
Based on the problems in the prior art, the invention takes the traditional Chinese herbal medicine Scutellarin (Scutellarin) as an amino acid template, modifies the RGD peptide structure of the GP IIb/IIIa receptor, improves the GP IIb/IIIa receptor specificity of the RGD peptide, improves the solubility of Scutellarin and develops a platelet aggregation resistant peptide mimetic compound.
Detailed Description
For a better understanding of the present invention, the following detailed description is given in conjunction with the following examples and drawings, but is not limited to the following examples.
The materials and reagents used in the following examples represent the meanings:
amide Resin: initial resin for polypeptide synthesis
Fmoc-linker: fmoc-linker, also known as 4- [ (2, 4-dimethoxyphenyl) (Fmoc-amino) methyl ] phenoxyacetic acid
DMF: n, N-dimethylformamide
DCM: methylene dichloride
DIC: diisopropylcarbodiimide
Ac2O: acetic anhydride
DIPEA: diisopropylethylamine
piperidine: piperidine derivatives
HOBt: 1-hydroxybenzotriazole
MeOH: methanol
TFA: trifluoroacetic acid
And (3) TIS: tri-isopropyl silane
ACN: acetonitrile
DMSO, DMSO: dimethyl sulfoxide
Trp: tryptophan
Boc: tert-butoxycarbonyl group
Asp: aspartic acid
OtBu: tert-butoxy radical
Gly: glycine
And (3) HomoArg: homoarginine
Pbf: 2,2,4,6, 7-pentamethyldihydrobenzofuran-5-sulfonyl
Pro: proline
Example 1 antiplatelet peptidomimetic Compound Scutellarin-HomoArg-Gly-Asp-Trp-NH2Synthesis of (2)
1.1 preparation of Fmoc-Linker-Amide Resin
50g (60mmol) of Amide Resin was weighed into a 2L solid phase synthesis reaction column. The resin was washed once with 400ml DMF and 500ml DCM was used to swell the resin for 20min and the solvent was removed by suction. The resin was washed 3 times with 400ml DMF.
80.9g (150mmol) of Fmoc-linker and 180 g (180mmol) of HOBt24.3g were weighed into a dry 500ml wide-mouth flask. Adding about 150ml DMF solvent to dissolve, placing in ice water bath to cool for 2min, adding DIC 30ml (195mmol) to activate for 3min to avoid water vapor. Adding the activated amino acid into the swelled resin for reaction for 2h, and pumping the reaction solution. The resin was washed 3 times with 400ml DMF for 2min each time.
Addition of Ac2The mixture of O28 ml (300mmol), DIPEA 10.7ml (60mmol) and DMF 400ml was sealed for 3h, and the resin was washed four times with DMF 500ml and twice with DCM 500 ml. The solvent was pumped off.
1.2 preparation of Fmoc-Trp (Boc) -Linker-Amide Resin
Fmoc-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-Trp (Boc) -OH 46g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. DIC 20ml was added and activated for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
1.3 preparation of Fmoc-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin
Fmoc-Trp (Boc) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-Asp (OtBu) -OH 36g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
1.4 preparation of Fmoc-Gly-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin
Fmoc-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
26g (87.5mmol) of Fmoc-Gly-OH and 14g (105mmol) of HOBt were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF was added to the solution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
1.5 preparation of Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin
Fmoc-Gly-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-HomoArg (Pbf) -OH 57.9g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction.
1.6 preparation of Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Linker-AmideResin
Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 600ml DMF and 2 times with 600ml DCM. The solvent was pumped off.
80.9g (175mmol) of Scutellarin and HOBt28g (210mmol) were weighed into a dry 500ml wide-mouth triangular flask. About 150ml of DMF was added to the solution. Cooling in ice water bath. 40ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 4 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction. MeOH 500ml, 300ml, 200ml shrink resin three times. And (5) vacuum pumping to dry. 125.8g were weighed. Drying and storing at low temperature.
1.7 Scutellarin-HomoArg-Gly-Asp-Trp-NH2Preparation of
Weighing Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Linker-Amide Resin dried to constant weight, adding 754ml lysate (TFA: TIS: H)2O) cleavage reaction for 2.5 h. After the reaction was completed, the resin was separated by filtration using a sand-core funnel. The filtrate was spin dried to 1/2 volume, excess ether (about 1.8L) was added until no more solids precipitated in solution, and the crude peptide of interest (56% pure) was obtained after filtration through a sand-core funnel and washing 5 times with ether. The mixture was dried in a vacuum desiccator to a constant weight of 56g in total.
1.8 refining
And (4) preparing and purifying by using a high performance liquid chromatograph.
Pretreatment: 56g Scutellarin-HomoArg-Gly-Asp-Trp-NH2The crude product was dissolved in 320ml of 70% aqueous acetic acid and filtered through a filter having a pore size of 0.45. mu.m.
Purification preparation conditions are as follows:
fluidity A: ACN; and (3) fluidity B: 0.5% aqueous acetic acid;
detection wavelength: 215 nm;
(1) elution gradient: purifying once (10-10% A) for 5min- (10-30% A) for 25min- (30-37% A) for 55min- (37-10% A) for 56min
A chromatographic column: c18-50 x 150 mm;
collecting conditions: the purity is more than 90% and 70-80%;
(2) performing secondary purification on the part with purity of more than 90%, wherein the elution gradient is (8-8% A)5min- (8-20% A)35min- (20-37%) 40min- (37-37% A)45min- (37-8% A)46 min;
a chromatographic column: c18-20 x 150 mm;
collecting conditions: the purity of the main peak is 99.3%.
Concentrating the main peak, and freeze-drying to obtain light yellow freeze-dried powder. (yield 30%, purity 99.1%)
Through mass spectrum detection, the target compound Scutellarin-HomoArg-Gly-Asp-Trp-NH2Was successfully synthesized. The molecular weight of the target compound is 990.
FIG. 1 shows the target compound Scutellarin-HomoArg-Gly-Asp-Trp-NH2(formula C)45H51N9O17) Mass spectrogram
Example 2 antiplatelet peptidomimetic Compound Scutellarin-HomoArg-Gly-Asp-NH2Synthesis of (2)
2.1 preparation of Fmoc-Linker-Amide Resin
50g (60mmol) of Amide Resin was weighed into a 2L solid phase synthesis reaction column. The resin was washed once with 400ml DMF and 500ml DCM was used to swell the resin for 20min and the solvent was removed by suction. The resin was washed three times with 400ml DMF.
80.9g (150mmol) of Fmoc-linker and 180 g (180mmol) of HOBt24.3g were weighed into a dry 500ml wide-mouth flask. Adding about 150ml DMF solvent to dissolve, cooling in ice water bath for 2min, adding 30ml DIC (195mmol) to activate for 3min to avoid water vapor. Adding the activated amino acid into the swelled resin for reaction for 2h, and pumping the reaction solution. The resin was washed 3 times with 400ml DMF for 2min each time.
Addition of Ac2The mixture of O28 ml (300mmol), DIPEA 10.7ml (60mmol) and DMF 400ml was sealed for 3h, and the resin was washed four times with DMF 500ml and twice with DCM 500 ml. The solvent was pumped off.
2.2 preparation of Fmoc-Asp (OtBu) -Linker-Amide Resin
Fmoc-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-Asp (OtBu) -OH 36g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
2.3 preparation of Fmoc-Gly-Asp (OtBu) -Linker-Amide Resin
Fmoc-Asp (OtBu) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for a first 5min and a second 8 min. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
26g (87.5mmol) of Fmoc-Gly-OH and 14g (105mmol) of HOBt were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF was added to the solution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
2.4 preparation of Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Linker-Amide Resin
Fmoc-Gly-Asp (OtBu) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-HomoArg (Pbf) -OH 57.9g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction.
2.5 preparation of Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Linker-Amide Resin
Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 600ml DMF and 2 times with 600ml DCM. The solvent was pumped off.
80.9g (175mmol) of Scutellarin and HOBt28g (210mmol) were weighed into a dry 500ml wide-mouth flask. About 150ml of DMF was added to the solution. Cooling in ice water bath. 40ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 4 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction. MeOH 500ml, 300ml, 200ml shrink resin three times. And (5) vacuum pumping to dry. Drying and storing at low temperature.
2.6 Scutellarin-HomoArg-Gly-Asp-NH2Preparation of
Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Linker-AmideResin peptide resin dried to constant weight is weighed, and 754ml of lysate (TFA: TIS: H)2O) cleavage reaction for 2.5 h. After the reaction was completed, the resin was separated by filtration using a sand-core funnel. And (3) spin-drying the filtrate to 1/2 volume, adding excessive ethyl ether (about 1.8L) until no solid is separated out in the solution, filtering by using a sand core funnel, and filtering and washing by using ethyl ether for 5 times to obtain the target crude peptide. Drying in a vacuum drier to constant weight.
2.7 refining
And (4) preparing and purifying by using a high performance liquid chromatograph.
Pretreatment: Scutellarin-HomoArg-Gly-Asp-NH2The crude product was dissolved in 320ml of 70% aqueous acetic acid and filtered through a filter having a pore size of 0.45. mu.m.
Purification preparation conditions are as follows:
fluidity A: ACN; and (3) fluidity B: 0.5% aqueous acetic acid;
detection wavelength: 215 nm;
(1) elution gradient: purifying for one time (10-10% A) for 5min- (10-30% A) for 25min- (30-37% A) for 55min- (37-10% A) for 56 min;
a chromatographic column: c18-50 x 150 mm;
collecting conditions: the purity is more than 90% and 70-80%;
(2) performing secondary purification on the part with purity of more than 90%, wherein the elution gradient is (8-8% A)5min- (8-20% A)35min- (20-37%) 40min- (37-37% A)45min- (37-8% A)46 min;
a chromatographic column: c18-20 x 150 mm;
collecting conditions: the purity of the main peak is 99.1%.
Concentrating the main peak, and freeze-drying to obtain light yellow freeze-dried powder. (yield 28%, purity 98.9%)
Example 3 antiplatelet peptidomimetic Compound Scutellarin-HomoArg-Gly-Asp-Trp-Pro-NH2Synthesis of (2)
3.1 preparation of Fmoc-Linker-Amide Resin
50g (60mmol) of Amide Resin was weighed into a 2L solid phase synthesis reaction column. The resin was washed once with 400ml DMF and 500ml DCM was used to swell the resin for 20min and the solvent was removed by suction. The resin was washed three times with 400ml DMF.
80.9g (150mmol) of Fmoc-linker and 180 g (180mmol) of HOBt24.3g were weighed into a dry 500ml wide-mouth flask. Adding about 150ml DMF solvent to dissolve, cooling in ice water bath for 2min, adding 30ml DIC (195mmol) to activate for 3min to avoid water vapor. Adding the activated amino acid into the swelled resin for reaction for 2h, and pumping the reaction solution. The resin was washed 3 times with 400ml DMF for 2min each time.
Addition of Ac2The mixture of O28 ml (300mmol), DIPEA 10.7ml (60mmol) and DMF 400ml was sealed for 3h, and the resin was washed four times with DMF 500ml and twice with DCM 500 ml. The solvent was pumped off.
3.2 preparation of Fmoc-Pro-Linker-Amide Resin
Fmoc-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
29.5g (87.5mmol) of Fmoc-Pro-OH and 14g (105mmol) of HOBt were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF was added to the solution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
3.3 preparation of Fmoc-Trp (Boc) -Pro-Linker-Amide Resin
Fmoc-Pro-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF, first for 5min and second for 8 min. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-Trp (Boc) -OH 46g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
3.4 preparation of Fmoc-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin
Fmoc-Trp (Boc) -Pro-Linker-Amide Resin was Fmoc-stripped twice with 500ml of 20% piperidine/DMF for 5min for the first time and 8min for the second time. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-Asp (OtBu) -OH 36g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
3.5 preparation of Fmoc-Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin
Fmoc-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
26g (87.5mmol) of Fmoc-Gly-OH and 14g (105mmol) of HOBt were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF was added to the solution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 500ml DMF for 2min each time, 2 times with 400ml DCM, and the solvent was removed by suction.
3.6 preparation of Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin
Fmoc-Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 500ml DMF and 2 times with 500ml DCM. The solvent was pumped off.
Fmoc-HomoArg (Pbf) -OH 57.9g (87.5mmol) and HOBt 14g (105mmol) were weighed into a dry 500ml wide-mouth flask. About 100ml of DMF solvent was added for dissolution. Cooling in ice water bath. 20ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 2 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction.
3.7 preparation of Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-AmideResin
Fmoc-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin was Fmoc-removed twice with 500ml of 20% piperidine/DMF for 5min for the first reaction and 8min for the second reaction. The resin was washed 4 times with 600ml DMF and 2 times with 600ml DCM. The solvent was pumped off.
80.9g (175mmol) of Scutellarin and HOBt28g (210mmol) were weighed into a dry 500ml wide-mouth flask. About 150ml of DMF solvent was added to dissolve. Cooling in ice water bath. 40ml DIC was added for 3min to avoid water vapor. Adding the activated amino acid into the deprotected resin to react for 2h, and pumping the reaction solution. The resin was washed 4 times with 600ml DMF for 2min each time, 2 times with 500ml DCM, and the solvent was removed by suction. MeOH 500ml, 300ml, 200ml shrink resin three times. And (5) vacuum pumping to dry. Drying and storing at low temperature.
3.8 Scutellarin-HomoArg-Gly-Asp-Trp-Pro-NH2Preparation of
Weighing Scutellarin-HomoArg (Pbf) -Gly-Asp (OtBu) -Trp (Boc) -Pro-Linker-Amide Resin peptide Resin dried to constant weight, adding 754ml lysate (TFA: TIS: H)2O) cleavage reaction for 2.5 h. After the reaction was completed, the resin was separated by filtration using a sand-core funnel. And (3) spin-drying the filtrate to 1/2 volume, adding excessive ethyl ether (about 1.8L) until no solid is separated out in the solution, filtering by using a sand core funnel, and filtering and washing by using ethyl ether for 5 times to obtain the target crude peptide. Drying in a vacuum drier to constant weight.
3.9 refining
And (4) preparing and purifying by using a high performance liquid chromatograph.
Pretreatment: Scutellarin-HomoArg-Gly-Asp-Trp-Pro-NH2The crude product was dissolved in 320ml of 70% aqueous acetic acid and filtered through a filter having a pore size of 0.45. mu.m.
Purification preparation conditions are as follows:
fluidity A: ACN; and (3) fluidity B: 0.5% aqueous acetic acid;
detection wavelength: 215 nm;
(1) elution gradient: purifying for one time (10-10% A) for 5min- (10-30% A) for 25min- (30-37% A) for 55min- (37-10% A) for 56 min;
a chromatographic column: c18-50 x 150 mm;
collecting conditions: the purity is more than 90% and 70-80%;
(2) performing secondary purification on the part with purity of more than 90%, wherein the elution gradient is (8-8% A)5min- (8-20% A)35min- (20-37%) 40min- (37-37% A)45min- (37-8% A)46 min;
a chromatographic column: c18-20 x 150 mm;
collecting conditions: the purity of the main peak is 99.0%.
Concentrating the main peak, and freeze-drying to obtain light yellow freeze-dried powder. (yield 29%, purity 98.6%)
Example 4 screening of antiplatelet peptidomimetics in antiplatelet aggregation
4.1 purpose
Selecting and synthesizing several groups of scutellarin-RGD modified bodies, respectively labeled as 19#, 21#, and 25#, and screening for anti-platelet aggregation.
The structures of the modified bodies are as follows:
19#：-Har-G-D-NH2(ii) a The molecular formula is as follows: c34H41N7O16(ii) a Molecular weight: 804;
21#：-Har-G-D-W-P-NH2(ii) a The molecular formula is as follows: c50H58N10O18(ii) a Molecular weight: 1087;
25#：-Har-G-D-W-NH2(ii) a The molecular formula is as follows: c45H51N9O17(ii) a Molecular weight: 990;
wherein,scutellarin;
har: homoarginine;
g: glycine;
d: aspartic acid;
w: tryptophan;
p: proline.
4.2 test sample Condition
No. 19 ethanol is insoluble and dissolved into 10 by adopting DMSO-3M (i.e. mol/L, the same below) and then diluted by 10 times of physiological saline;
21#, 25#, Epti (Epfebutat, a disulfide bridged cyclic peptide) was dissolved in 50% ethanol to 10%-3M, diluting with normal saline by 10 times;
dissolving scutellarin for injection into 10% with distilled water-2M, and diluted with physiological saline accordingly.
4.3 animals
SD rats, SPF grade, male, 220-260 g, provided by Beijing Wintonlihua laboratory animal technology, Inc., animal production license number SCXK (Jing) 2012-0001.
4.4 instruments
Model 540 platelet aggregometer, available from CHRONO-LOG, USA.
4.5 method for measuring platelet aggregation function
Pentobarbital sodium is injected into the abdominal cavity for 60mg/kg anesthesia, the abdominal aorta is used for blood taking, 3.2 percent sodium citrate is used for anticoagulation, and the volume ratio of the whole blood to the anticoagulant is 9: 1. Platelet Rich Plasma (PRP) was prepared by centrifugation at 800rpm for 10min, and Platelet Poor Plasma (PPP) was prepared by centrifugation at 3000rpm for 10 min. The PRP is adjusted by PPP to keep the platelet count at 3.5-4.0 × 108one/mL.
Platelet aggregation was measured by turbidimetry. 215 mu L LPRP and 25 mu L of the tested medicine are added into a cuvette and put into a pre-heating hole of a platelet aggregation instrument for incubation for 10min at 37 ℃. The maximum aggregation function of platelets and the inhibition rate relative to the solvent control group were determined by adjusting 100% aggregation rate with PPP and 0% aggregation rate with PRP, and adding 10. mu.L of Adenosine Diphosphate (ADP) as an inducer with continuous stirring to a final concentration of about 2. mu.M, respectively.
4.6 results
The inhibition rate of the 19#, 21#, 25# scutellarin-RGD modified body relative to the solvent control group is measured by using a turbidimetric method, and is as follows:
TABLE 1 inhibition (%)
FIG. 2 inhibition ratio of each sample at different concentrations
From the above experimental results, the 25# sample shows a very strong anti-platelet aggregation function, suggesting that the use of scutellarin as an amino acid template can effectively fix the conformation of the RGD-like peptide, and achieve and exceed the efficacy of epti, and far exceed the efficacy of scutellarin. This compound is designated WK001 by the present invention.
Example 5 preliminary metabolism test of intravenous administration in rats
5.1 Experimental conditions:
instrument Agilent 1100 LC/MSD.
secondly, mass spectrum conditions comprise an ESI ionization mode, a negative ion detection mode,
capillary voltage 2000v, atomization gas pressure 55psi,
the drying air flow rate is 5mL/min, and the drying air temperature is 200 ℃.
thirdly, mass spectrum analysis, namely respectively carrying out WK001 and internal standard (rutin)MS analysis, selection [ M-H]-For ions used for WK001 monitoring, the mass to charge ratio was m/z988 and the internal standard mass to charge ratio was m/z 609.
chromatographic conditions, wherein the chromatographic column is a Zorbax XDB C18 column (150X 4.6mm, 5 mu m), the mobile phase is acetonitrile-water-formic acid (20:80:0.1), the flow rate is 0.5mL/min, and the sample injection amount is 20 mu L.
5.2 sample collection:
WK001 (a 2mg/mL physiological saline solution, with an administration volume of 0.5mL) was administered intravenously to rats and males at a dose of 1 mg/mouse, approximately 1 mL of blank blood was taken as a blank blood sample before administration, and blood was taken at 1min, 5min, 10min, 20min, 30min, 1h, 2h, and 4h after administration.
5.3 sample treatment:
numbering centrifuge tubes at different times in advance, adding 200 μ L acetonitrile, weighing, recording, collecting blood about 100 μ L, adding into 200 μ L acetonitrile, vortexing for 1min, weighing and correcting acetonitrile amount (to make its ratio be 2: 1), centrifuging, collecting supernatant 200 μ L, adding into another centrifuge tube, adding 50 μ L internal standard solution, vortexing, and placing in 37 deg.C water bath N2Blow-dry and dissolve the residue in 150. mu.L of mobile phase.
5.4 measurement results:
the results of the blood concentration change of the selected peptidomimetic compound at different times are determined as follows:
TABLE 2 plasma drug concentration changes at different times
FIG. 3 the change of blood concentration of WK001 at different times
The results indicate the half-life (t) of WK0011/2) At 4.9min, no metabolites were found by full scan analysis.
Drawings
FIG. 1 shows the target compound Scutellarin-HomoArg-Gly-Asp-Trp-NH2(formula C)45H51N9O17) Mass spectrum, [ M-H ]]-m/z988.38 is a compound molecular ion peak, and m/z 493.69 is a compound double-charge molecular ion peak.
FIG. 2 shows the inhibition of each sample at different concentrations.
Fig. 3 the change of blood concentration of WK001 at different times.
Claims (10)
1. A peptide-like compound with the general formula (I) has the following structure
2. The peptide, AA, according to claim 11: selected from the group consisting of Arg, Har, Lys, Orn;
AA2: selected from Trp, Ser, Val, Phe(iii) a group consisting of, or absent;
AAm: selected from Pro, or absent;
x is selected from the group consisting of: OH, NH2，OR1，
Wherein R is1Selected from fatty alkyl groups.
3. The peptide according to claim 1, characterized in that: AA1is-L-Har, AA2And AAmIs absent, X is NH2。
4. The peptide according to claim 1, characterized in that: AA1is-L-Har, AA2is-L-Trp, AAmIs Pro and X is NH2。
5. The peptide according to claim 1, characterized in that: AA1is-L-Har, AA2is-L-Trp, AAmIs absent, X is NH2。
6. A process for obtaining peptidomimetics of general formula (I) according to any one of claims 1 to 5, characterized in that it is carried out in a solid phase.
7. Use of at least one peptide according to any one of claims 1 to 5 in a pharmaceutical composition for the treatment of cardiovascular diseases.
8. The use according to claim 7, wherein the treatment is anti-platelet aggregation.
9. Pharmaceutical composition, characterized in that it comprises a peptidomimetic of general formula (I) according to any one of claims 1 to 5.
10. The pharmaceutical composition according to claim 9, wherein the peptidomimetic of formula (I) is contained in a pharmaceutically acceptable sustained release system or carrier selected from the group consisting of liposomes, nanocapsules, microcapsules, nanocapsules, cysts, micelles, nanospheres, microspheres, nanospheres, lipospheres, microemulsions, nanoemulsions, nanoparticles, microparticles and nanoparticles.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109453124A (en) * | 2018-11-23 | 2019-03-12 | 深圳市维琪医药研发有限公司 | A kind of lamp-dish flower acetic lyophilized preparation and preparation method thereof of RGD class peptide modification |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991004746A1 (en) * | 1989-09-29 | 1991-04-18 | Rhone-Poulenc Rorer International (Holdings) Inc. | Anti-thrombotic peptides and pseudopeptides |
| CN1132515A (en) * | 1993-09-30 | 1996-10-02 | 新日本制铁株式会社 | Novel peptide, active as inhibitors of platelet aggregation |
| CN1392157A (en) * | 2002-07-10 | 2003-01-22 | 牛勃 | New anti-embolism medicine using small peptide arginine-glycine-aspartic acid (RGD) as leading compound |
| US20030216481A1 (en) * | 2002-04-30 | 2003-11-20 | Unigen Pharmaceuticals, Inc. | Formulation of a mixture of Free-B-ring flavonoids and flavans as a therapeutic agent |
| CN1657042A (en) * | 2004-02-18 | 2005-08-24 | 车庆明 | Application of erigeron breviscapus aglucon in preparating medicine |
| CN101485629A (en) * | 2008-01-16 | 2009-07-22 | 沈阳药科大学 | Drug delivery system and preparation method thereof |
| CN103768047A (en) * | 2014-02-19 | 2014-05-07 | 天津中医药大学 | Application of scutellarein to preparing medicine for preventing and/or treating thrombotic diseases |
| CN104940187A (en) * | 2015-06-19 | 2015-09-30 | 昆药集团股份有限公司 | New application of scutellarin |
| CN105198849A (en) * | 2015-08-20 | 2015-12-30 | 南京中医药大学 | Scutellarein amine derivative and preparation method and application thereof |
-
2018
- 2018-05-07 CN CN201810426114.0A patent/CN108610394B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991004746A1 (en) * | 1989-09-29 | 1991-04-18 | Rhone-Poulenc Rorer International (Holdings) Inc. | Anti-thrombotic peptides and pseudopeptides |
| CN1132515A (en) * | 1993-09-30 | 1996-10-02 | 新日本制铁株式会社 | Novel peptide, active as inhibitors of platelet aggregation |
| US20030216481A1 (en) * | 2002-04-30 | 2003-11-20 | Unigen Pharmaceuticals, Inc. | Formulation of a mixture of Free-B-ring flavonoids and flavans as a therapeutic agent |
| CN1392157A (en) * | 2002-07-10 | 2003-01-22 | 牛勃 | New anti-embolism medicine using small peptide arginine-glycine-aspartic acid (RGD) as leading compound |
| CN1657042A (en) * | 2004-02-18 | 2005-08-24 | 车庆明 | Application of erigeron breviscapus aglucon in preparating medicine |
| CN101485629A (en) * | 2008-01-16 | 2009-07-22 | 沈阳药科大学 | Drug delivery system and preparation method thereof |
| CN103768047A (en) * | 2014-02-19 | 2014-05-07 | 天津中医药大学 | Application of scutellarein to preparing medicine for preventing and/or treating thrombotic diseases |
| CN104940187A (en) * | 2015-06-19 | 2015-09-30 | 昆药集团股份有限公司 | New application of scutellarin |
| CN105198849A (en) * | 2015-08-20 | 2015-12-30 | 南京中医药大学 | Scutellarein amine derivative and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 宋树霖: "灯盏乙素衍生物的合成及生物活性评价", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 操锋等: "灯盏乙素酯类前药的合成、理化性质及降解研究", 《药学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109453124A (en) * | 2018-11-23 | 2019-03-12 | 深圳市维琪医药研发有限公司 | A kind of lamp-dish flower acetic lyophilized preparation and preparation method thereof of RGD class peptide modification |
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