CN106214674B - The medical usage of 7- hydroxyls-butylphenyl phthaleine - Google Patents
The medical usage of 7- hydroxyls-butylphenyl phthaleine Download PDFInfo
- Publication number
- CN106214674B CN106214674B CN201610538809.9A CN201610538809A CN106214674B CN 106214674 B CN106214674 B CN 106214674B CN 201610538809 A CN201610538809 A CN 201610538809A CN 106214674 B CN106214674 B CN 106214674B
- Authority
- CN
- China
- Prior art keywords
- butylphenyl phthaleine
- hydroxyls
- disease
- cerebral
- phthaleine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Abstract
The invention belongs to field of pharmaceutical chemistry technology, and in particular to 7- hydroxyls-butylphenyl phthaleine is preparing the application in preventing and/or treating cardiac and cerebral vascular diseases drug.
Description
Technical field
The invention belongs to field of pharmaceutical chemistry technology, and in particular to 7- hydroxyls-butylphenyl phthaleine is preparing prevention and/or the treatment heart
Application in cerebrovascular drug.
Background technology
Cardiovascular and cerebrovascular disease be endanger human life with health common disease and frequently-occurring disease, be the three big cause of death of the mankind it
One, wherein ischemic cerebrovascular disease accounts for prodigious ratio.Acute ischemic stroke has high incidence, high disability rate, high death
The characteristics of rate, brings greatly pain even life danger to patient.
Thrombus, oxidativestress damage are the cardiovascular and cerebrovascular diseases such as acute ischemic cerebral apoplexy and coronary heart disease and myocardial infarction
Etiological.The underlying pathology of Arterial thrombosis is mainly that atherosclerotic plaque ulceration causes blood platelet at ulceration
Stick and assembles and then start coagulation process, and arterial thrombus caused by atherosclerosis may occur in which artery blood supply once being formed
The ischemic at position and necrosis, as myocardial infarction, ischemic cerebral apoplexy are medium;Candelario-Jalil E, Curr Opin
Investig Drugs, 2009,10 (7): 644-654].Numerous studies show oxidativestress damage neuron after cerebral ischemia
It plays a crucial role in damage.Under normal circumstances, the generation and removing of internal oxygen radical are balances.When vivo oxidation and resist
Oxidation is unbalance, and body or tissue are in the state of oxidation, causes neutrophil leucocyte inflammatory infiltration, protease secretion to increase, production
Raw a large amount of intermediate oxidation products, and tissue damage or disease is caused to be oxidative stress.When cerebrum ischemia, especially fill again
When note, considerably beyond the Scavenging activity of own endogenous antioxidant system, excessive free radical can cause for the generation of free radical
The peroxidating of lipid, protein, nucleic acid, and coup injury cell, lead to meronecrosis or apoptosis, so free radical and oxidation
It stress play an important role in ischemic neuronal cellular damage.The enzyme that human body removes free radical mainly has 3 classes：Superoxides discrimination
Change enzyme (SOD), catalase (CAT), glutathione peroxidase (GSH-Px).SOD and GSH-Px is important in vivo
Oxygen free radical scavenger reflects tissue anti-oxidative damage ability.SOD, can be effective by disproportionated reaction scavenging activated oxygen
Prevent oxidative damage, blocking oxide from stress play important protective effect to oxidative damage in the signal transduction after cerebral ischemia；GSH-
Px is located in mitochondria, can be catalyzed H2O2It is reduced into H2O.Malonaldehyde (malonaldehyde, MDA) is body lipid peroxidating
The final metabolite of reaction organizes the metabolism status of oxygen radical in the horizontal antimers indirectly of MDA, also reflects tissue by certainly
Degree is attacked by base.
Currently, preventing the drug of cerebral arterial thrombosis, cerebral infarction, brain edema few in number in clinic, wherein calcium overload is
The key factor of Neuron Apoptosis process, can cause neuronal death, and calcium antagonist Nimodipine clinical research has been proved pair
Cerebral ischemia has certain neuroprotection and reminiscence to act on, but Ca2+Interior stream is happened at the early stage of cerebral ischemia, therefore calcium ion is short of money
The application of anti-agent has certain limitation.And damage caused by being generated for cerebral ischemia and Reperfu- sion phase a large amount of oxygen radicals
Evil, Edaravone have radicals scavenging and antioxidation, be listed at present it is few in number acute for treating
Apoplexy drug.The limitation of the medicine is can only to realize the clinical application of acute stage, and have hemorrhage trans formation and light, moderate
The side reactions such as kidney, hepatic disorder.It is 3- butylphthalides (abbreviation butylphenyl phthaleine, trade name that another, which is applied to clinical drug,
" En Bipu "), it has antioxidant radical, the effect of antithrombotic, anti-platelet aggregation, Chinese patent ZL93117148.2 public affairs
The activity of anti-cerebral ischemia is opened, but the medicine is only used for treating light, moderate acute ischemic cerebral apoplexy, and has had generation nausea, fash
And the toxic side effects such as mental symptom.
Therefore, for the treatment of cerebral ischemia diseases such as cerebral apoplexy, cerebral infarction, brain edema, current existing drug cannot
Meets the needs of clinical, the clinical lower new drug of urgently better efficacy, toxicity occurs.
The present invention is in carrying out anti-oxidant emergent, antiplatelet condensation and the research of anti-cerebral ischemia disease medicament, from one plant of fox
Isolated three compounds in excrement mould fungal bacterial strain：7- hydroxyls-butylphenyl phthaleine, 4- hydroxyls-butylphenyl phthaleine and 7- methoxyl groups-butylbenzene
Phthalein.The inside and outside drug effect and toxicity research result that the present invention carries out prove：7- hydroxyls-butylphenyl phthaleine, 4- hydroxyls-butylphenyl phthaleine and 7- first
Oxygroup-butylphenyl phthaleine all has the drug actions such as anti-oxidant, antithrombotic and anti-cerebral ischemia；7- hydroxyls-butylphenyl phthaleine compared with butylphenyl phthaleine,
Make with drug effects such as lower dosage, more notable, excellent anti-platelet aggregation, anti-oxidation stress and anti-cerebral ischemia activity
With, and there is lower toxicity, that is, there is higher drug safety；4- hydroxyls-butylphenyl phthaleine has similar with butylphenyl phthaleine
The drug actions such as anti-platelet aggregation, anti-oxidation stress and anti-cerebral ischemia activity；7- methoxyl groups-butylphenyl phthaleine has certain anti-blood
Platelet aggregation, anti-oxidation stress drug action, but the drug action of the compound is slightly below butylphenyl phthaleine.
7- hydroxyls-butylphenyl phthaleine be foreign Rhizoma Chuanxiong (Cnidium officinaleMakino) the chemical composition of volatile oil（US
2006/0246157）With the Dai Xiechanwu [ of fungi fox excrement mould;Heterocycles, 1998, 48(9): 1931-1933；
CN201510113631.9 ].The compound is found to have prevention and treatment diabetes（US20090192218）, depression and
The illness related with impaired neurotransmission such as anxiety（US20100184852）And with the relevant disease of angiogenesis
（US20060246157）.But by literature search, the compound of the present invention there are no anti-oxidant, antithrombotic and treating cerebral ischemia
Report.What is more important is compared with analogue butylphenyl phthaleine, and the compound of the present invention rises with lower drug
Dosage is imitated, the feature of more superior drug action and lower toxic reaction under same dose.So the invention reside in
A kind of novel medical use of 7- hydroxyls-butylphenyl phthaleine is provided, which is related to the compound and combinations thereof in cardiac-cerebral ischemia disease
The prevention and/or treatment of disease, cardiovascular and cerebrovascular circulation disturbances, thrombus etc., it is characterised in that more safe and effective to clinical offer
Prevent and treat drug.
Invention content
The purpose of the present invention is provided with the medicinal usage for preventing and/or treating cardiac and cerebral vascular diseases for clinic.
The present inventor has found that 7- hydroxyls-butylphenyl phthaleine is preparing treatment and/or preventing cardiac and cerebral vascular diseases through numerous studies
Application in drug.
The application, wherein cardiac and cerebral vascular diseases are ischemic cardiovascular and cerebral vascular disease, thrombus disease, cardiovascular and cerebrovascular circulation disturbances.
The application, wherein coronary heart disease caused by the ischemic cardiovascular and cerebral vascular disease refers to ischemic, brain edema, cerebral apoplexy,
Brain blood flow lesion, cerebral infarction disease.
The application, wherein the thrombus disease is cerebral thrombus, deep vein thrombosis, pulmonary embolism disease.
The pharmaceutical composition of 7- hydroxyls-butylphenyl phthaleine provided by the invention is preparing prevention and/or treatment cardiac and cerebral vascular disease
There is same application in medicine.
The application, wherein cardiac and cerebral vascular diseases are ischemic cardiovascular and cerebral vascular disease, thrombus disease and cardiovascular and cerebrovascular circulation disturbances.
The application, wherein coronary heart disease caused by the ischemic cardiovascular and cerebral vascular disease refers to ischemic, brain edema, cerebral apoplexy,
Brain blood flow lesion, cerebral infarction disease.
The application, wherein the thrombus disease is cerebral thrombus, deep vein thrombosis, pulmonary embolism disease.
The present invention provides 7- hydroxyls-butylphenyl phthaleine and its pharmaceutical composition, since the notable medicine under low dosage may be implemented
Effect acts on, and significantly improves the protecting effect to ischemic brain damage, reduces the risk of adverse reaction generation.
7- hydroxyls-butylphenyl phthaleine provided by the invention and its pharmaceutical composition show lower toxic reaction under high dose,
Safer, efficient protective agents application is provided.
The compound of the present invention has following excellent performance：
In anti-platelet aggregation experiment, 7- hydroxyls-butylphenyl phthaleine has more superior anti-platelet aggregation drug action.With
Published butylphenyl phthaleine is compared, and the compound of the present invention shows external anti-platelet aggregation more excellent drug action,
Show that reach the drug dose that the identical the compounds of this invention that uses of inhibition platelet aggregation effect uses lower；In phase
With under dosage, the compound of the present invention is to platelet aggregation inhibiting rate higher.It is more aobvious to illustrate that 7- hydroxyls-butylphenyl phthaleine of the present invention has
The antithrombotic drug action of work.
In oxidation resistance experiment, 7- hydroxyls-butylphenyl phthaleine has more superior antioxidation.With published butylbenzene
Phthalein is compared, and the compound of the present invention oxidation resistance shows more excellent drug action, that is, is shown and reached identical anti-
Oxidation activity includes antioxidant radical, improves superoxide dismutase（SOD）, catalase (CAT), glutathione peroxidating
Object enzyme（GSH-Px）Level reduces the compounds of this invention agent used in the effect of malondialdehyde (MDA) content
Amount is lower, and under same dose, the compound of the present invention has stronger oxidation resistance.Illustrate 7- hydroxyls-fourth of the present invention
Phthalide has more significant anti-oxidative stress.
In to brain in the influence experiment of the Range of Cerebral Infarction of focal ischemia type brain damage model, 7- hydroxyls-butylphenyl phthaleine is aobvious
It shows to the significant protective effect of brain tissue impairment caused by obstruction of artery.Compared with butylphenyl phthaleine, the compounds of this invention protection is big
The more excellent drug action of brain tissue impairment caused by brain Middle cerebral artery occlusion, can reach when being in particular in using relatively low-dose
Butylphenyl phthaleine reduces the infarct size of similar Middle cerebral artery thrombosis model rat under high dose, i.e. the compound of the present invention has
Smaller conspicuousness dosage；Under lower dosage, butylphenyl phthaleine does not show to cause the protection of cerebral injury to make cerebral ischemia
With, but the compounds of this invention 7- hydroxyls-butylphenyl phthaleine can express out the pharmacodynamic results of conspicuousness, illustrate the compounds of this invention to heart and brain
Ischemic disease has more superior preventive and therapeutic action.
In acute toxicity test, the compound of the present invention 7- hydroxyls-butylphenyl phthaleine and butylphenyl phthaleine（Trade name " En Bipu "）
Compare, 7- hydroxyls-butylphenyl phthaleine than butylphenyl phthaleine have higher maximum tolerated dose and aobvious toxic dose, so the present invention be prevention or
Improve cardiovascular and cerebrovascular row disease, especially prevention or improvement heart and brain cycle, prevention and treatment ischemic cardiovascular and cerebral vascular disease provides one kind
More safely and effectively treatment means and method.
The pharmaceutical composition of the present invention refers to that the compounds of this invention containing therapeutically effective amount is active ingredient, and is contained
One or more pharmaceutically acceptable carriers.
Pharmaceutically acceptable carrier described above refers to the pharmaceutical carrier of pharmaceutical field routine（Also referred to as auxiliary material）, wherein
The common auxiliary material of solid pharmaceutical preparation includes filler such as starch, sucrose etc.；Adhesive for example cellulose derivative, alginates, gelatin and
Polyvinyl pyrrolidone；Wetting agent such as ethyl alcohol, water；Disintegrant such as starch and its derivative such as sodium carboxymethyl starch（CMS-
Na）, low-substituted hydroxypropyl cellulose（LS-HPC）, crosslinked polyvinylpyrrolidone（PVPP）, cross-linked carboxymethyl cellulose receives
（CMC-Na）Deng and gas-producing disintegrant；Sorbefacient such as quaternary ammonium compound；Surfactant such as hexadecanol；Absorption carries
Body such as kaolin or soap clay；Lubricant such as talcum powder, calcium stearate or magnesium；In addition other auxiliary can also be added in the composition
Agent such as flavouring agent, sweetener etc..
The present invention about oral medication, compound can be and pharmaceutically acceptable carrier knot well known in the art
It closes.This kind of carrier enables the compounds of this invention to prepare piece agent, pill, pastille, capsule, liquid, gel, syrup, slurry
Liquid, suspension etc., for being taken orally by institute patient to be treated.The capsule that the pharmaceutical preparation that can be taken orally includes, is made of gelatin,
And soft fluid sealant wafer, it is made of gelatin and a kind of plasticizer, such as glycerine.Capsule can contain active constituent under
The mixture of row ingredient：Filler, such as lactose；Adhesive, such as starch；And/or lubricant, such as talcum powder or stearic acid
Magnesium or superfine silica gel powder；With optional stabilizer.In soft capsule, reactive compound can be dissolved or suspended in suitable liquid
In body, such as fat oil, atoleine or liquid macrogol.Furthermore it is possible to which stabilizer is added.All oral administration preparations
The dosage of this kind of administration should be all suitable for.Pharmaceutical composition can also include the solid being suitble to or gel phase carriers or figuration
Agent.The example of this kind of carrier or excipient includes but not limited to calcium carbonate, calcium phosphate, various sugar, starch, cellulose derivative,
Gelatin, polymer, polymer are, for example, polyethylene glycol.
The auxiliary material of liquid preparation includes water for injection, physiological saline and pH conditioning agents such as sodium hydroxide, also other
Auxiliary material such as isotonic regulator etc., those skilled in the art can rationally select according to the liquid preparation customary preparation methods of pharmaceutical field
Common supplementary product kind and dosage are selected, final products is made to meet conventional IV agent or freeze-dried requirement used for intravenous injection.
Aqueous injection suspensions can contain the substance for increasing suspension viscosity, such as sodium carboxymethylcellulose glucan；Optionally, suspension is also
Suitable stabilizer can be contained or increase the reagent, such as hydroxy propyl-Beta cyclodextrin etc. of compound solubility, it is molten to increase
Xie Xing prepares highly enriched solution.Alternatively, active constituent can be powder type, suitable carrier regeneration, example are being used using preceding
Such as sterile pyrogen-free water.
The compound of the present invention or pharmaceutical composition, which can be used for preparing, prevents and/or treats ischemic cardiovascular and cerebral vascular disease, anti-blood
The drug of bolt, improvement cardiovascular and cerebrovascular circulation disturbances etc..Its various dosage form can be by those skilled in the art, according to the routine of pharmaceutical field
It is prepared by production method.Such as active constituent is made to be mixed with one or more carriers, then it is made into required dosage form, including piece
Agent, capsule, granule, water-soluble salt can be by those skilled in the art, according to the quiet of pharmaceutical field in the compounds of this invention
It is freeze-dried etc. that intravenous injection, intravenous injection is made in arteries and veins injection conventional production process.
Preferred formulation form of the present invention is tablet, capsule or intravenous injection.
Specific implementation mode
With reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
Experiment material as used in the following examples is SILVER REAGENT, 7- hydroxyls-butylphenyl phthaleine, 4- hydroxyls-butylphenyl phthaleine unless otherwise specified
It is self-control sample with 7- methoxyl groups-butylphenyl phthaleine, butylphenyl phthaleine is Enbipu Pharmacy Co., Ltd., Shiyao Group.'s product.
The preparation of 1. 7- hydroxyls of embodiment-butylphenyl phthaleine, 4- hydroxyls-butylphenyl phthaleine and 7- methoxyl groups-butylphenyl phthaleine
The preparation referenced patent of 7- hydroxyls-butylphenyl phthaleine, 4- hydroxyls-butylphenyl phthaleine and 7- methoxyl groups-butylphenyl phthaleine
CN201510113631.9, by fox excrement penicillium bacterial strain NCC3421（It has been preserved in China General Microbiological culture presevation management
The heart, deposit number are CGMCC No.9094）It is inoculated in the fermentation tank equipped with 30L fermentation mediums, 26 DEG C of tank temperature, 220
Rpm is stirred, and cultivates 120 h.4000 rpm of gained culture centrifuges 20min, collects mycelia, processing industry alcohol steep, and filtering obtains
To filtrate.Above-mentioned filtrate is diluted with water to concentration of alcohol 40%, with equipped with 2L D312 resins pillar adsorb, then respectively with
40%, 55%, 70% ethanol elution, branch receive, and merge the part containing purpose thing, and 40 DEG C of vacuum distillations stop when to appear muddy
Only, it is stored at room temperature, purpose thing crude product is drained to obtain in filtering.
Above-mentioned crude product is dissolved with methanol, and distilled water is added dropwise to micro- aobvious muddiness.Warm makes solution clarify, and stands, and crystal is slow
It is precipitated, puts 4 DEG C overnight, filter, vacuum drying obtains 36.0 g of 7- hydroxyls butylphenyl phthaleine, content 99.5%.Disposing mother liquor, in upper ODS
Compression leg（4.9*50 cm, C18,30 μm）Separation, 10% ~ 50% acetonitrile-water（Containing 0.5% acetic acid）Linear gradient elution collects master
Part before and after peak and main peak, main peak are 7- hydroxyls-butylphenyl phthaleine；After main peak pre-impurity and post-impurity merge, carries out HPLC and prepare [Chromatography
Column：Receive micro- Unisil C18 30.0*300 mm, mobile phase：30% ~ 60% acetonitrile-water (containing 0.5% acetic acid) 50min, flow velocity
35.0 ml/min], obtain 4- hydroxyls-butylphenyl phthaleine (Rt:28.6min, 520mg), 7- hydroxyls-butylphenyl phthaleine (Rt:30.9min,
846mg), 7- methoxyl groups-butylphenyl phthaleine (Rt:33.1min, 615mg).
ESI-MS and the NMR data ownership of above three compound are as follows：
4- hydroxyls-butylphenyl phthaleine：ESI-MS m/z 207.2[M+H]+, 1H-NMR(500MHz, CDCl3) 7.33(1H,
dd, J=7.5, 1.5, H-5), 7.31(1H, t, J=7.5, H-6), 7.10 (1H, dd, J=7.5, 1.5, H-
7), δ5.52(1H, dd, J=8.0, 3.0, H-3), 2.31(1H, m, H-1′a), 1.73(1H, m, H-1′b),
1.37(4H, m, H-2′, H-3′), 0.89 (3H, t, J=7.0, H-4′), 9.59(1H, s, 4-OH)。13C-NMR
(125MHz, CDCl3) δ171.2(C-1), 80.9(C-3), 136.1(C-3a), 152.4(C-4), 120.2(C-5),
130.3(C-6), 115.9(C-7), 127.8(C-7a), 32.3(C-1′), 26.9(C-2′), 22.4(C-3′), 13.9
(C-4′).It is consistent with EP1932527 report data.
7- hydroxyls-butylphenyl phthaleine：ESI-MS m/z 207.2[M+H]+, 1H-NMR(500MHz, CDCl3) δ5.49(1H,
dd, J=8.0, 3.0, H-3), 7.53(1H, t, J=8.0, H-5), 6.89(1H, dd, J=8.0,1.5, H-6),
6.91 (1H, dd, J=8.0,1.5, H-4), 2.02(1H, m, H-1’a), 1.78(1H, m, H-1’b), 1.45
(2H, m, H-2’), 1.38 (2H, m, H-3′), 0.89 (3H, t, J=7.0, H-4′), 7.81(1H, s, 7-
OH)。13C-NMR(125MHz, CDCl3) δ172.3(C-1), 83.1(C-3), 150.6.1(C-3a), 115.4(C-4),
137.0(C-5), 113.2(C-6), 156.6(C-7), 111.4(C-7a), 34.4(C-1′), 27.0(C-2′), 22.5
(C-3′), 14.0(C-4′).With Heterocycles, 1998,48 (9):The data of 1931-1934 reports are almost the same.
7- methoxyl groups-butylphenyl phthaleine：ESI-MS m/z 221.1[M+H]+, 1H-NMR(500MHz, CDCl3) δ 7.61
(1H, t, J=8.0, H-5), 6.96 (1H, d, J=7.5, H-4), 6.93(1H, d, J=8.0, H-6), 5.39
(1H, dd, J=8.0, 4.0, H-3), 4.00 (3H, s, 7-OCH3), 2.02 (1H, m, H-1′a), 1.72
(1H, m, H-1′b), 1.45 (2H, m, H-2′), 1.38 (2H, m, H-3′), 0.90 (3H, t, J = 7.5,
H-4′);。13C-NMR(125MHz, CDCl3) δ168.7 (C-1), 158.5 (C-7), 153.0 (C-3a), 136.1
(C-5), 113.6 (C-7a), 113.3 (C-4), 110.5 (C-6), 80.2 (C-3), 55.9 (7-OCH3),
34.4 (C-1′), 26.7 (C-2′), 22.4 (C-3′), 13.8 (C-4′).With Planta Medica, 2008,74
(1):The data of 69-72 reports are consistent.
2. external platelet aggregation inhibitory activity of embodiment
（1）Experimental method：Platelet aggregation inhibitory activity is evaluated, using ADP as derivant, using Born turbidimetrys.
Wistar rats（Hebei Medical University's animal center, experimental animal quality certification number：1304152）Eye socket takes blood, with 3.8%
Sodium citrate anti-freezing (whole blood and anti-coagulants volume ratio 9: 1), centrifuges (500~800 rpm, 10 min) at room temperature, prepares rich
Collect thrombocyte plasma (PRP), after isolating PRP, then centrifuges (3000 rpm, 15 min), prepare platelet poor plasma
(PPP), with PPP tune 0.Take PRP (200 μ L) that the 5 μ L of DMSO solution of the target compound of equimolar concentration, solvent is added
Control group adds isometric DMSO, after incubating 2 min, using 10 μ Μ ADP as derivant, by Born turbidimetry for Determination blood platelets
Aggreation.External platelet aggregation inhibitory activity measurement is carried out to target compound, calculates platelet aggregation (AR)
Inhibit percentage (AIR) with aggregation.It is calculated according to the following formula and assembles and inhibit percentage：Platelet aggregation=[(PRP
Absorbance adds ADP to assemble absorbance)/PRP Xi Guangdus ]× 100%, Platelet aggregation inhibitor percentage=[(solvent control group
Aggregation rate administration group aggregation rate)/solvent control group Ju Jishuai ]×100%.
（2）Experimental result：
Compared with ADP induced coagulation groups, * P<0.05, **<0.01
The result of study of the present invention is shown, compared with the platelet aggregation group of ADP inductions, 7- hydroxyls-butylphenyl phthaleine drug 30
μ Μ, 10 μ Μ, 3 μ Μ and 1 μ Μ obviously inhibit platelet aggregation (P<0.05 or P<0.01), and there is good dose-effect
Relationship.Compared with butylphenyl phthaleine group, at 30 μ Μ, 10 μ Μ same doses, anti-coagulation rate of the 7- hydroxyls-butylphenyl phthaleine to blood platelet
It is all higher than butylphenyl phthaleine；At 3 μ Μ and 1 μ Μ, the inhibitory activity that 7- hydroxyls-butylphenyl phthaleine has still had, and at this concentration
Butylphenyl phthaleine unrestraint activity；Anticoagulation under each concentration of 4- hydroxyls-butylphenyl phthaleine is substantially less than 7- hydroxyls-butylphenyl phthaleine, with
Butylphenyl phthaleine is substantially close.7- methoxyl groups-butylphenyl phthaleine does not show apparent anticoagulation in the 30 following concentration of μ Μ concentration.
Illustrate that the compound of the present invention 7- hydroxyls-butylphenyl phthaleine has more superior anti-platelet aggregation effect compared with other compounds.
3. antioxidant activity in vitro of embodiment
(1) experiment material and method
The measurement of total reducing power：The phosphoric acid that 0. 2 mol/L pH 6. 6 are added in the sample of 1 mL various concentrations is slow
Rush the potassium ferricyanide 2.5 mL, 50 DEG C of 20 min of water-bath of 2.5 mL of solution and mass fraction 1%.After ice bath rapid cooling, it is added
10% solution of trichloroacetic acid 2. 5 mL, 3000 r/min centrifuge 10 min, take 1. 0 mL of supernatant, and distilled water 1.0 is added
0. 1% FeCl of mL and mass fraction30. 2mL, mixing are stored at room temperature 10 min, and absorbance is measured at 700 nm of wavelength
Value measures reducing power according to absorbance value, and calculates reducing power %.
Remove the ability of ultra-oxygen anion free radical：Take the sample of different volumes that 50 mM pH values 8. 2 are added respectively
Tris-HCl buffer solutions are configured to the solution of various concentration.Above-mentioned solution is placed in 25 DEG C of waters bath with thermostatic control and keeps the temperature 20 min,
0. 05 mL, the 50 mM pyrogallol solution of 25 DEG C of preheatings is added, shakes up rapidly, every 0. 5 at 325 nm of wavelength
Min measures absorbance value 1 time, and linear session is 4 min, and the absorbance value of measurement is A1.Mouse thymus cells group, with phase
Same volume distilled water replaces polyphenol extracting solution, absorbance value to be denoted as A2.Inhibiting rate (%)=(Δ A2/Δ T- Δs A1/Δ T)/
(ΔA2 /ΔT) ×100。
The measurement of anti-lipid peroxidation (LPO) effect：10 mL centrifuge tubes are taken, 0. 2 mL 1 are added：25 is diluted
Yolk suspension (yolk is formulated with the phosphate buffer of isometric pH 7. 4, with before need use magnetic agitation 10
Min), then it is separately added into 2.0 mL of polyphenol extracting solution and 25 mM FeSO of various concentration4·7H20. 2 mL of O solution, is placed in 37
30 min are vibrated in DEG C water bath with thermostatic control.0. 85% thio bar of 0.5 mL, 20% solution of trichloroacetic acid and 1.0 mL are added after taking-up
Than appropriate acid, mixing is placed in 100 DEG C of water-baths and keeps the temperature 20 min, cooling, and 15 min are centrifuged under 3000 rpm, take 3.0 mL supernatants
Liquid measures the absorbance value at 532 nm of wavelength.Free radical inhibiting rate (%)=(blank absorbency-sample liquid absorbance)/sky
White absorbance × 100.
(2) experimental result
The present invention researches show that：The antioxidant activity that 7- hydroxyls-butylphenyl phthaleine and 4- hydroxyls-butylphenyl phthaleine have all had, it is bright
It is aobvious to be higher than butylphenyl phthaleine；But the oxidation resistance of 7- hydroxyls-butylphenyl phthaleine is higher than 4- hydroxyls-butylphenyl phthaleine.Illustrate the compound of the present invention
7- hydroxyls-butylphenyl phthaleine has more superior antioxidation.
Influence and oxidation resistance of the embodiment 4. to local rats with cerebral ischemia cerebral infarct volume
(1) experiment material and method
Experimental animal：Wistar rats, weight 250-280g.Animal is stablized 1 week after buying, and keeps diet, drinking-water
Normal and circadian rhythm is normal.
The preparation of evaluating focal brain ischemia in rats：Middle cerebral artery occlusion is prepared using internal carotid line brush
（Middle cerebral artery occlusion, MCAO) cerebral ischemia re-pouring model.7% three chloroethenes of hydration of animal
After aldehyde (6 mL/kg) anesthesia, prone position is fixed on operating table, sterilizes skin, neck midsection, and separation right side neck always moves
Arteries and veins, external carotid artery, internal carotid, gently remove vagus nerve, ligature and cut external carotid artery, follow internal carotid forward, ligation
Wing arm artery.Folder closes arteria carotis communis proximal part, makees a kerf from the distal end of the ligature of external carotid artery, and it is 0.285 to include into outer diameter
The nylon wire of mm enters internal carotid by arteria carotis communis bifurcated, (from bifurcated until being then inserted into light resistance slowly
Locate about 20 mm), it blocks all blood supplies of arteria cerebri media, after 2.0 h of right side cerebral ischemia, gently extracts nylon wire, restore blood
For carrying out Reperfu- sion, skin suture, disinfection.
Animal packet and administration：Animal is grouped at random, i.e. sham-operation group, model group, butylphenyl phthaleine control group, different pharmaceutical
Administration group.Only, every group of 10-13 of other each groups is only by sham-operation group 5-6.
Administration route and the frequency：Intravenously administrable is administered once through tail vein injection immediately after Reperfu- sion, and interval is after 2 hours
It is administered once, is administered 2 times altogether again.
Detection：It after cerebral ischemia 24 hours, takes blood and puts to death animal, take brain, dye, calculate brain infarction area.It is dyed through TTC
Afterwards, normal structure dye deeply takes on a red color, and infarction tissue is white.The infarct size of calculation every is sought, and finally superposition is converted into infraction
Volume.Infarct volume indicates with the percentage of shared cerebral hemisphere, cerebral infarct volume (%)=(operation contralateral hemisphere volume-hand
The volume of the non-infarcted portion of art side hemisphere)/operation contralateral hemisphere volume * 100%.3000 rpm of taken blood plasma is centrifuged 10 minutes,
Serum is taken, for oxidation index parametric measurement in serum.Measure according to the method for kit measure MDA contents and SOD in serum,
CAT and GSH-Px activity.
（2）Test result：
Sample see the table below 2 to the protective effect of local rats with cerebral ischemia cerebral infarct volume and internal anti-oxidation stress effect
With table 3.
With sham-operation group ratio ##<0.01；With model group ratio, * P<0.05, **<0.01
The results show that postoperative for 24 hours in addition to sham-operation group is without focus of infarct, model group and administration group rat have in various degree
Focus of infarct.Compared with model group, 7- hydroxyls-butylphenyl phthaleine 10-1.0 mg/kg processing group rat cerebral infarction volumes obviously subtract
Few, infraction degree is obviously improved (P<0.05 or P<0.01), and there is apparent dose-effect relationship.Importantly, and butylphenyl phthaleine
It compares, 7- hydroxyls-butylphenyl phthaleine is 1 mg/kg to the improvement result effective dose that MCAO rat cerebral tissues block, and butylphenyl phthaleine
Effective dose illustrates that the improvement result that 7- hydroxyls-butylphenyl phthaleine blocks rats with cerebral ischemia brain tissue is stronger in 10 mg/kg.It says
Bright the compound of the present invention has smaller conspicuousness dosage, i.e., under lower dosage, the compound of the present invention can table
Reveal conspicuousness effect.
With sham-operation group ratio##P<0.01；With model group ratio * P<0.05, **<0.01
The results show that the degree of oxidation of postoperative 24 h cerebral infarction models groups is above sham-operation group.Compared with model group, 7- hydroxyls
The content of antioxidase SOD, CAT, GSH-Px in base-butylphenyl phthaleine 10-1.0 mg/kg processing group rat blood serums are above
Model group (P<0.05 or P<0.01), and lipid peroxide MDA contents are obviously reduced, and with good dosage according to
The relationship of relying；Compared with butylphenyl phthaleine group, under same dose, the antioxidase content of 7- hydroxyls-butylphenyl phthaleine is above butylphenyl phthaleine group,
And MDA contents are less than butylphenyl phthaleine processing group, butylphenyl phthaleine group does not show anti-oxidative stress under less than 10 mg/kg dosage.
Illustrate that the compound of the present invention has smaller conspicuousness dosage, i.e., at lower doses, the compound of the present invention
Show conspicuousness anti-oxidation stress effect.
5. Acute toxicity of embodiment is tested
（1）Experiment material and method
Intravenously administrable：It is small using single or 24 with 30% cyclodextrin 7- hydroxyls-butylphenyl phthaleine and butylphenyl phthaleine to 5 mg/mL
When interior multiple intravenously administrable mode investigate acute toxicity of the test sample to SD rats, excipient group rat gives equivalent volumes
Excipient.Type：SD rats, SPF grades, each group quantity：6（It is male）.
（2）Experimental result：
The above test result shows that the toxicity of the compounds of this invention 7- hydroxyls-butylphenyl phthaleine is significantly lower than butylphenyl phthaleine, illustrates it
With drug safety advantage more significant than butylphenyl phthaleine.
Claims (3)
1.7- hydroxyls-butylphenyl phthaleine is preparing the application in preventing and/or treating cardiac and cerebral vascular diseases drug；Wherein cardiovascular and cerebrovascular
Property disease be ischemic cardiovascular and cerebral vascular disease, thrombus disease, cardiovascular and cerebrovascular circulation disturbances.
2. application according to claim 1, wherein coronary heart disease, brain water caused by the ischemic cardiovascular and cerebral vascular disease refers to ischemic
Swollen, cerebral apoplexy, brain blood flow lesion, cerebral infarction disease.
3. application according to claim 2, wherein the thrombus disease is cerebral thrombus, deep vein thrombosis, pulmonary embolism disease
Disease.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610538809.9A CN106214674B (en) | 2016-07-11 | 2016-07-11 | The medical usage of 7- hydroxyls-butylphenyl phthaleine |
| PCT/CN2016/103066 WO2018010332A1 (en) | 2016-07-11 | 2016-10-24 | Medical use of 7-hydroxy-butylphthalide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610538809.9A CN106214674B (en) | 2016-07-11 | 2016-07-11 | The medical usage of 7- hydroxyls-butylphenyl phthaleine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106214674A CN106214674A (en) | 2016-12-14 |
| CN106214674B true CN106214674B (en) | 2018-10-26 |
Family
ID=57520536
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610538809.9A Active CN106214674B (en) | 2016-07-11 | 2016-07-11 | The medical usage of 7- hydroxyls-butylphenyl phthaleine |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN106214674B (en) |
| WO (1) | WO2018010332A1 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112625016A (en) * | 2019-09-24 | 2021-04-09 | 华北制药集团新药研究开发有限责任公司 | 7-hydroxy butylphthalide crystal form B and preparation method thereof |
| CN115243686B (en) | 2020-03-20 | 2024-04-09 | 石药集团恩必普药业有限公司 | Use of butylphthalide and derivatives thereof |
| CN113666895B (en) * | 2020-05-15 | 2023-12-08 | 华北制药集团新药研究开发有限责任公司 | Halogenated 2-benzo [ c ] furanone compounds and application thereof |
| CN114762700B (en) * | 2021-01-13 | 2023-03-21 | 浙江泛亚生物医药股份有限公司 | Application of penicillium fox dung culture |
| CN113024422B (en) * | 2021-03-12 | 2022-12-20 | 上海科州药物研发有限公司 | Butylphthalide ring-opening compound, pharmaceutical compound, and preparation methods, compositions and applications thereof |
| CN115141861A (en) * | 2022-08-03 | 2022-10-04 | 华北制药集团新药研究开发有限责任公司 | Method for producing 7-hydroxy butylphthalide by fermenting penicillium fox dung |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7482027B2 (en) * | 2005-04-30 | 2009-01-27 | Angiolab, Inc. | Composition for the prevention or treatment of diseases associated with angiogenesis |
-
2016
- 2016-07-11 CN CN201610538809.9A patent/CN106214674B/en active Active
- 2016-10-24 WO PCT/CN2016/103066 patent/WO2018010332A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7482027B2 (en) * | 2005-04-30 | 2009-01-27 | Angiolab, Inc. | Composition for the prevention or treatment of diseases associated with angiogenesis |
Non-Patent Citations (1)
| Title |
|---|
| 丁苯酞脑缺血治疗作用的相关药理学研究进展;端木寅等;《中药药理与临床》;20121231;第28卷(第3期);126-129 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2018010332A1 (en) | 2018-01-18 |
| CN106214674A (en) | 2016-12-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106214674B (en) | The medical usage of 7- hydroxyls-butylphenyl phthaleine | |
| JP6133415B2 (en) | Production method and use of hydroxyl safflower yellow A sodium | |
| US6737085B2 (en) | Apocynum venetum extract for use as antidepressant | |
| JP2018503637A (en) | Biphenyl derivatives and uses thereof | |
| KR970011555B1 (en) | Green tea extract im antithrombotic agent and a process for preparing the same | |
| KR20080107794A (en) | Composition comprising the extract of arctium lappa seed for anti-cancer | |
| JP2018508479A (en) | Use of biphenols in the preparation of drugs to prevent and treat ischemic stroke | |
| JPH0114206B2 (en) | ||
| CN105367582B (en) | Bilobalide B derivates and its application in drug | |
| KR100265385B1 (en) | Extract of rhodiola sp. for prevending and treating circulating diseases | |
| JP6523634B2 (en) | Thrombus preventive and therapeutic agent and method for producing the same | |
| DK2855487T3 (en) | Compounds for the treatment of ischemia-reperfusion-related diseases | |
| JP2008115089A (en) | Agent for prevention and treatment of ischemic disease and organ preservation agent | |
| KR101788658B1 (en) | New hydroxysafflor yellow a salt for pharmaceutical formulation | |
| CN112220847A (en) | Fresh pomelo flesh water extract powder, preparation, antithrombotic effect and application thereof | |
| JP6627141B2 (en) | Method for preparing safflower buckwheat extract, extract prepared thereby, and use of extract | |
| CN110237085A (en) | Pharmaceutical preparation and application thereof | |
| CN110237086A (en) | Composition, pharmaceutical preparation and purposes | |
| KR101973907B1 (en) | Phamaceutical composition for preventing or treating of specific cancers containing extracts, fractions or sargachromenol from Sargassum serratifolium | |
| KR101923769B1 (en) | Phamaceutical composition for preventing or treating of specific cancers containing extracts, fractions or sargachromenol from Sargassum serratifolium | |
| KR100504966B1 (en) | Antihypertensive constituents of ginseng saponin | |
| CN105646493B (en) | It is a kind of to be used to prevent and treat compound of organ damage and its production and use | |
| KR100962984B1 (en) | A Pharmaceutical Composition for Treating or Preventing Cancer Comprising Ubiquinone-9 or Pharmaceutically Acceptable Salt Thereof | |
| Khosravi et al. | Biological Evaluation of New Oxadiazole-Based Synthetic α-glycosidase Inhibitors for Hyperglycemia Management: A Research Study: New Oxadiazoles act as a-glycosidase inhibitor | |
| CN112220827A (en) | Mulberry leaf water extract powder, preparation, antithrombotic effect and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |