CN104840478A - Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases - Google Patents
Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases Download PDFInfo
- Publication number
- CN104840478A CN104840478A CN201510086547.2A CN201510086547A CN104840478A CN 104840478 A CN104840478 A CN 104840478A CN 201510086547 A CN201510086547 A CN 201510086547A CN 104840478 A CN104840478 A CN 104840478A
- Authority
- CN
- China
- Prior art keywords
- nadph
- medicine
- disease
- application
- sclerosis
- 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.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
-
- 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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
Abstract
The invention discloses application of NADPH in preparation of drugs used for treating or preventing cardio-cerebrovascular diseases. Research results show that NADPH can protect vascular endothelial cells and maintain normal vascular permeability and is a candidate drug used for treating or preventing cardio-cerebrovascular diseases, especially thrombotic diseases, hypertension, coronary heart diseases, vascular sclerosis and vascular sclerosis caused by damage of vascular endothelial cells.
Description
Technical field
The present invention relates to the new indication of NADPH, the specifically application of NADPH in preparation treatment or prevention cardiovascular and cerebrovascular diseases medicament.
Background technology
Cardiovascular and cerebrovascular disease refers to that cardiovascular and cerebrovascular disease are referred to as.The feature that cardiovascular and cerebrovascular disease has " sickness rate is high, mortality rate is high, disability rate is high, relapse rate is high, complication many " (" four is high by more than one "), having become the heavy burden of global health care and health resources at present, is the dead enemy of second time health revolution.At present, China's Patients with Cardiovascular/Cerebrovascular Diseases is more than 2.7 hundred million people, and after especially China enters senescence society, the sickness rate of cardiovascular and cerebrovascular disease is still in continuous increase, therefore, study the pathomechanism of cardiovascular and cerebrovascular disease and treat the vital task that protection is the world of medicine always.Cardiac-cerebral ischemia pipe disease incidence mechanism is very complicated, and the mechanism of causing a disease that research cardiac-cerebral ischemia pipe disease is new, the target spot that searching drug effect is new, the exploitation for the prevention and therapy medicine of cardiovascular and cerebrovascular disease has the meaning of important reality.
NADPH (triphosphopyridinenucleotide, NADPH) produced through phosphopentose pathway (PPP) metabolism by glucose, as electron donor of paramount importance in cell and biosynthetic reducing agent, can be reproducibility biosynthesis and hydrion is provided.NADPH is the coenzyme of the reductase of glutathion (GSH), and oxidized form of glutathione (GSSG) can be made to generate reduced form GSH, maintains the normal contents of the GSH of reduced form.GSH is antioxidant important in cell, and some protein containing sulfydryl, fat and proteases can be protected from the destruction of oxidant, especially at the integrity important role maintaining erythrocyte membrane.NADPH is except participating in the biosynthesis of cholesterol, fatty acid, monooxygenase system, steroid hormone etc., and the biology also participating in hydroxylation reaction and medicine, poisonous substance and some hormone in body transforms.Such as, NADPH can utilize the electron donor separating poison cell, reduces organism oxidized form compound, maintain its redox balance, play a significant role in oxidant defense system by internal metabolism.NADPH also can enter respiratory chain by means of the 1-Hydroxy-1,2,3-propanetricarboxylic acid. effect of shuttling back and forth and produce ATP: because mitochondrial inner membrane is very low to the permeability of material, it is oxidized that the NADPH that mitochondrion produces outward directly can not enter respiratory chain.H on NADPH can be given NAD+ under the effect of Isocitrate dehydrogenase, then enters respiratory chain produce power by NAD+.The maintenance of cellular energy metabolism and minimizing ROS (reactive oxygen species) are to cells survival, most important to organizing of hypoxic-ischemic especially, generally believe that energy metabolism impairment and oxidative stress are the important mechanisms of cardiac-cerebral ischemia diseases, research shows to increase cellular energy metabolism ability, reduces cell ROS and produces the cell injury that can alleviate hypoxic-ischemic and cause.Based on the multiple physiological function of NADPH, NADPH is applied in the treatment of some disease by more existing researchs at present.Such as, NADPH disclosed in Chinese patent literature CN103340890A is as the preparation of the application preventing and treating cerebral infarction medicine aspect, utilize the antioxidation treatment cerebral infarction of NADPH, due to the main mechanism that oxidation emergency is cerebral ischemia, and NADPH is endogenous anti-oxidative material, therefore gives exogenous NADPH and there is wind action in well treatment cerebral ischemia.At present, whether not yet clear and definite NADPH is applicable to the treatment of other diseases.
Summary of the invention
Technical problem to be solved by this invention is to provide the application of the new indication of a kind of NADPH, i.e. the application of NADPH in preparation treatment or prevention cardiovascular and cerebrovascular diseases medicament.
The invention provides the application of NADPH in preparation treatment or prevention cardiovascular and cerebrovascular diseases medicament.
Further, described cardiovascular and cerebrovascular disease refers to the one in thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis.
Further, described thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis refer to thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis that vascular endothelial cell damage causes.
Described thrombotic disease refers to the one in arterial thrombus, phlebothrombosis.
Present invention also offers NADPH in preparation protection vascular endothelial cell, the application safeguarded in the medicine of the normal permeability of blood vessel.
Described medicine comprises the NADPH of pharmaceutical effective amount and pharmaceutically acceptable carrier.
The medicine being used for the treatment of or preventing cardiovascular and cerebrovascular disease provided by the invention, described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.
The medicine being used for the treatment of or preventing any one disease in thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis provided by the invention, described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.Further, described thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis refer to the thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, the atherosclerosis that are caused by vascular endothelial cell damage.
Provided by the invention for the protection of vascular endothelial cell, the medicine safeguarding the normal permeability of blood vessel; described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.
The active component of scheme of the present invention using NADPH as preparation treatment cardiovascular and cerebrovascular diseases medicament; confirm that NADPH has protection vascular endothelial cell, safeguards the effect of the normal permeability of blood vessel; be cardiovascular and cerebrovascular disease, especially treat thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerotic drug candidate.
Because NADPH is an endogenic antioxidant, do not find that there is toxic and side effects when treatment use, therefore, there is the advantage that consumption is little, safe.In addition, NADPH by oral, injectable, can pass through nasal mucosa and percutaneous drug delivery, taking convenience.Further, because oxidative stress and energy metabolism impairment are the Common Mechanisms of other organs and tissues ischemic injuries, therefore, NADPH also can have been widely used in Other diseases.
Accompanying drawing explanation
In order to make content of the present invention be more likely to be clearly understood, below in conjunction with accompanying drawing, the present invention is further detailed explanation, wherein,
Fig. 1 is the impact of external source NADPH on the primary culture endothelial cell HUVEC cell survival rate of low sugar anoxia;
Fig. 2 is that therapeutic gives the impact of NADPH on participating in the immunocyte of blood brain barrier in permanent cerebral infarction mouse brain;
Fig. 3 preventatively gives the impact of NADPH on cerebral ischemia Reperfu-sion apoplexy blood-brain barrier of mice permeability;
Fig. 4 is the impact that therapeutic gives NADPH and damages permanent cerebral infarction blood-brain barrier of mice; Fig. 4 a is Blood Brain Barrier (BBB) permeability measurement result; Fig. 4 b is blood-brain barrier permeability test result;
Fig. 5 is the impact that therapeutic gives NADPH and damages myocardial ischemia-reperfusion mouse cardiac muscle; Fig. 5 a is TTC coloration result; Fig. 5 b is the percentage ratio comparison diagram that ischemic infarction district myocardial mass accounts for ischemic region myocardial mass.
Detailed description of the invention
The capsule of embodiment 1 containing NADPH
The capsule of the present embodiment comprises following composition:
NADPH 20g; Suspending agent microcrystalline Cellulose 60g; The antiseptic tert-butyl group-4-hydroxyanisol 0.04g; Magnesium stearate lubricant 2g; Filler lactose adds to 200g.
Its preparation method comprises the following steps:
Take the NADPH of above-mentioned recipe quantity and each pharmaceutic adjuvant, mix homogeneously, cross 60 mesh sieve three times, incapsulate and get final product.
As in customary adjuvant used include but not limited in filler, disintegrating agent, lubricant, binding agent, correctives, suspending agent, antiseptic the mixture of one or more.
Specifically, described filler also can be replaced one or more the mixture in pregelatinized Starch, mannitol, chitin, microcrystalline Cellulose, sucrose;
Described disintegrating agent also can be replaced one or more the mixture in starch, polyvinylpolypyrrolidone, sodium carboxymethyl cellulose, carboxymethyl starch sodium;
Described lubricant also can be replaced one or more the mixture in Pulvis Talci, silicon dioxide, sodium lauryl sulphate;
Described suspending agent also can be replaced one or more the mixture in polyvinylpyrrolidone, sucrose, agar, hydroxypropyl emthylcellulose;
Described antiseptic also can be replaced one or more the mixture in parabens, benzoic acid, sodium benzoate, sorbic acid, sorbate;
Described binding agent also can be replaced one or more the mixture in polyvinylpyrrolidone, hydroxypropyl emthylcellulose;
Described correctives also can be replaced sweeting agent and/or essence; Described sweeting agent is one or more the mixture in saccharin sodium, Aspartane, sucrose, cyclamate;
Certainly, described customary adjuvant includes but not limited to the above-mentioned scope enumerated, and those skilled in the art can do adaptive selection and adjustment according to practical situation.
Those skilled in the art can use any mode as known in the art to use medicine of the present invention, include but not limited to external, oral, Sublingual, per nasal, parenteral, locally, subcutaneous, injection, percutaneous or rectum route of administration.The preferred buccal dosage forms of pharmaceutical composition of the present invention and injection type, oral cavity type is selected from oral liquid, capsule, effervescent tablet, oral medicine film or spray; Injection type is selected from injectable powder and aqueous injection etc. for muscle, subcutaneous administrations or intravenous drip.And medicine of the present invention can adopt method as known in the art to be made as corresponding dosage form.
Experimental example
Below, for verifying that technique effect of the present invention carries out following experiment:
The exogenous NADPH of experimental example 1 protects primary culture endothelial cell HUVEC and does
(1) experiment material:
Test the primary culture endothelial cell HUVEC cell used and be all purchased from U.S. ATCC cell bank.Conditions of cryopreservation: 2ml cryopreservation tube, often pipe 1,600,000 cell, containing 70% DMEM in high glucose, 20% domestic hyclone, 10%DMSO.
(2) experimental program:
Endotheliocyte HUVEC cultivates: condition of culture: 37 DEG C (5%CO2,95% air), saturated humidity, DMEM in high glucose culture medium, and culture medium often rises containing 100U penicillin and 100U streptomycin; 10% domestic hyclone; Treat that Growth of Cells merges to about 80-90%, go down to posterity with after the digestion of pancreas enzyme-EDTA liquid, go down to posterity density: 5 × 10
5/ bottle went down to posterity every 2-3 days; To take the logarithm the HUVEC cell of growth, add appropriate trypsin-EDTA Digestive system, attached cell is come off, collecting cell, counting, be made into cell suspension (5 × 10 with containing the culture fluid of 10% hyclone
4/ ml), in 96 orifice plates, every hole adds 100 μ l, 37 DEG C, 5%CO2 cultivates 24h.Get NADPH to be dissolved in physiological saline solution and to be mixed with the solution that mother solution is 10mmol/L, join mtt assay cell survival rate in cell culture medium after membrane filtration is degerming and detect:
Cytoactive detects: be inoculated in by cell in 96 orifice plates, is 1,5 until Growth of Cells to adding final concentration during the index stage, and 10,20, the NADPH of 40 μMs.Be negative control with solvent, put into incubator and continue to cultivate 48h under the same terms.Terminate to cultivate front 4h and add MTT (5mg/ml, D-Hanks ' dissolves) 10 μ l, supernatant is sucked during off-test, add 150 μ l DMSO, each hole OD value is measured in 570nm place with enzyme-linked immunosorbent assay instrument, get 6 hole OD values to average, calculate suppression ratio: suppression ratio (IR%)=(1-instrument connection OD/ control wells OD) × 100%.
(3) experimental result
Fig. 1 is the impact of external source NADPH on the primary culture endothelial cell HUVEC activity of low sugar anoxia.Compared with Normal group, 5,10, the NADPH effect 48h of 20 μMs of concentration can make HUVEC cell survival rate obviously reduce.The NADPH effect 48h cell survival rate of 5 μMs is about 65.3% (p < 0.05); 10 μMs of NADPH effect 48h cell survival rates are about 73.6% (p < 0.01); 20 μMs of NADPH effect 48h cell survival rates are about 70.9% (p < 0.01).1) matched group; 2) anoxia low sugar group; 3) anoxia low sugar group+NADPH1 μM; 4) anoxia low sugar group+NADPH5 μM; 5) anoxia low sugar group+NADPH10 μM; 6) anoxia low sugar group+NADPH20 μM; 7) anoxia low sugar group+NADPH40 μM.
*compared with matched group, p < 0.01;
#compared with anoxia low sugar group, p < 0.05;
##compared with anoxia low sugar group, p < 0.01.Wherein
*represent p<0.01,
#represent p<0.05,
##represent p<0.01.
After the preventative NADPH of giving of experimental example 2 alleviates brain injury, blood brain barrier is relevant immune
Cell effect
(1) experiment material
Cleaning grade male ICR mouse, quality 23 ~ 28g, University Of Suzhou's Experimental Animal Center provides, laboratory animal production licence number: XCYK (Soviet Union) 2002-2008, laboratory animal occupancy permit number: SYXK (Soviet Union) 2002-0037.C57BL6 and Tg (Itgax-Venus) 1Mnz system mice, body weight 22 ~ 27g, male.Tg (Itgax-Venus) 1Mnz system mice is MGI Products, and this Mus is that CD11c-eYFP is positive, participates in immunoreation situation after brain injury for studying dendritic cell.Room temperature 22 DEG C, humidity 50-60%, well-ventilated, manually round the clock (12h/12h), water of freely ingesting.Before experiment, male mice is adapted to 2d in feeding environment.Dextran-Texas (Red): Invitrogen Products, NADPH reagent is purchased from Jiangsu sigma Reagent Company; Synthetic, semi-synthetic can be passed through in the source of exogenous NADPH medicine, and biological extraction obtains.
(2) experimental program:
Permanent middle cerebral artery thromboembolism cerebral ischemic model is set up.Tg (Itgax-Venus) 1Mnz system mice, body weight 22 ~ 27g, male.Experiment is grouped into sham operated rats, cerebrovascular trauma model group, cerebrovascular trauma+NADPH (2.5mg/kg) treatment group, every treated animal 6.NADPH artificial cerebrospinal fluid dilutes, and adopts intracerebroventricular, mice tricorn administration 2 μ l (cerebrovascular trauma model group intracerebroventricular artificial cerebrospinal fluid).Mice adopts chloral hydrate anesthesia, fixing, neck median incision, and separation left common carotid, neck are interior, external carotid artery.The ligation of common carotid artery proximal part, in distance neck, inserts the unified nylon wire bolt of diameter, blocking blood flow 24h before external carotid artery bifurcated.Tail vein injection Dextran-40 solution during ischemia 22h.Through 2h body-internal-circulation, use 10% chloral hydrate anesthesia, open thoracic cavity, expose heart, by 10ml 10mmol/L PBS solution in heart left ventricle's perfusion, Dextran-Texas remaining in cerebral tissue is rinsed out.NADPH makes administration in first 2 hours in permanent middle cerebral artery thromboembolism cerebral ischemic model.The relevant immune cell responses of determination experiment animal blood brain barrier after permanent middle cerebral artery thromboembolism cerebral ischemic model 24h.
(3) experimental technique
Blood brain barrier is about the detection of immune cell responses: 22h mouse tail vein injection Dextran-Texas after cerebral microvascular damage, after 2h, laboratory animal broken end gets brain, the cerebral tissue of formalin perfusion fixation subsequently, vibratome is cut into slices, section is collected in PBS liquid, adopts floating method to carry out immunohistochemical staining to it.CD11c-eYFP antibody labeling dendron shape positive cell, DAPI carries out nuclear targeting, mounting.The distribution of reaction in striatum brain district of dendron shape inflammatory cell and expression around confocal microscopy blood vessel.
(4) experimental result
Fig. 2 preventatively gives the NADPH impact of reacting permanent cerebral infarction mice local immune response.Sham-operated control group mouse brain district is dispersed in and is distributed with CD11c-eYFP cell, and cell space is less, and dendritic arbors is clear.After cerebral ischemia 24 hours, in Brain striatal region, all visible significantly CD11c-eYFP reactivity increased model group, and cell space distortion increases.NADPH (2.5mg/kg) organizes the CD11c-eYFP reactivity that ventricles of the brain administration effectively can reduce stratium regions.NADPH has well protection blood brain barrier effect, reacts relevant with adjustment local immunity
The preventative NADPH that gives of embodiment 3 reduces the experiment of cerebral ischemia re-pouring Blood Brain Barrier (BBB) permeability
(1) experiment material is with experimental example 2.
(2) experimentation
1) mice transience middle cerebral artery occlusion model is set up:
Get ICR normal mouse group, body weight 23 ~ 28g, random point 2 groups, often organize 20, be divided into normal saline group (vehicle group), NADPH (7.5mg/kg) dosage group; NADPH 1 week (every day injects 2 times) before ischemia is injected in vivo by tail vein.Adopt internal carotid artery line brush, improve preparation mice right ischemia MCAO model a little, mice is with 4% chloral hydrate (400mg/kg) intraperitoneal injection of anesthesia, and we adopt line brush to prepare ischemia model, and separation neck is total, neck outer and internal carotid artery, outer and the total proximal part of neck of ligation strength, line bolt (6023, Doccol Corporation, Redlands, USA) insert from neck until anterior cerebral artery initiating terminal, block middle cerebral artery blood supply.After blocking blood flow 2h, pull out Outlet bolt and realize Reperfu-sion.Sham operated rats mice is except not plug wire, and all the other steps are all identical with treatment group with ischemia group.In whole operation process, room temperature remains on 22 ~ 25 DEG C, adopts automatic temperature control heating pad by the temperature control of mice anus at 37 ± 0.5 DEG C.Postoperatively animal is placed in the raising box being placed with clean bedding and padding, freely drinks water, take food.
2) permanent middle cerebral artery thromboembolism cerebral ischemic model is set up.
C57BL6 system mice, body weight 22 ~ 27g, male.Experiment is grouped into sham operated rats, cerebrovascular trauma model group, cerebrovascular trauma+NADPH (2.5mg/kg) treatment group, every treated animal 6.NADPH artificial cerebrospinal fluid dilutes, and adopts intracerebroventricular, mice tricorn administration 2 μ l (cerebrovascular trauma model group intracerebroventricular artificial cerebrospinal fluid).Mice adopts chloral hydrate anesthesia, fixing, neck median incision, and separation left common carotid, neck are interior, external carotid artery.The ligation of common carotid artery proximal part, in distance neck, inserts the unified nylon wire bolt of diameter, blocking blood flow 24h before external carotid artery bifurcated.Tail vein injection Dextran-40 solution during ischemia 22h.Through 2h body-internal-circulation, use 10% chloral hydrate anesthesia, open thoracic cavity, expose heart, by 10ml10mmol/L PBS solution in heart left ventricle's perfusion, Dextran-Texas remaining in cerebral tissue is rinsed out.NADPH makes administration in first 2 hours in permanent middle cerebral artery thromboembolism cerebral ischemic model.Determination experiment animal Blood Brain Barrier (BBB) permeability after permanent middle cerebral artery thromboembolism cerebral ischemic model 24h.
(3) experimental technique
1) blood-brain barrier permeability test: inject after 2% azovan blue (EB) normal saline solution (4ml/kg) 1h through tail vein after cerebral ischemia re-pouring 23h and mice broken end is got brain, weigh and put into 50% solution of trichloroacetic acid, homogenate and centrifugal (10000rpm, 20min), go supernatant according to the ratio ethanol dilution of 1:3.OD value is detected at 620nm place; Calculate EB content in cerebral tissue.
2) Blood Brain Barrier (BBB) permeability measures: 22h mouse tail vein injection Dextran-Texas after cerebral microvascular damage, after 2h, laboratory animal broken end gets brain, the cerebral tissue of formalin perfusion fixation subsequently, and vibratome is cut into slices, DAPI carries out nuclear targeting, mounting.The Dextran-Texas fluorescence detected under laser confocal microscope in the cerebrovascular surrounding brain parenchyma in cerebral ischemia brain district is strong and weak.
3) data statistics and analysis: data all represent with mean ± standard error (Mean ± SEM), statistical analysis adopts one factor analysis of variance (one-way ANOVA), and p<0.05 is that significant difference has significance.
(4) experimental result
Fig. 3 preventatively gives the impact of NADPH on cerebral ischemia re-pouring apoplexy blood-brain barrier of mice permeability.Compared with vehicle group, 1 week (tail vein 2 times every day) significantly reduces the blood-brain barrier of mice permeability (p<0.01) of cerebral infarction mice after giving NADPH in advance.
*represent p<0.01.
Fig. 4 preventatively gives the impact of NADPH on permanent cerebral infarction blood-brain barrier of mice permeability.Fig. 4 a is Blood Brain Barrier (BBB) permeability measurement result, and Fig. 4 b is blood-brain barrier permeability test result.24h after cerebral ischemia, model group compares sham operated rats all visible significantly the spilling of red fluorescence dyestuff Dextran-Texas of cerebral cortex and stratium regions.NADPH (2.5mg/kg) organizes ventricles of the brain administration effectively can reduce spilling of the Dextran-Texas of ischemia induction cerebral cortex and stratium regions.Prompting NADPH shows well protection blood brain barrier effect.
Embodiment 4 therapeutic gives NADPH and reduces treating myocardial ischemia damage
(1) experiment material
Adult male SD rats, body weight 270 ~ 350g, cleaning grade.Thered is provided by University Of Suzhou's medical board Experimental Animal Center.[credit number: (Soviet Union) SYXK2007-0035].Azovan blue (Evans Blue, EB, sigma) gets SD normal rat group, body weight 23 ~ 28g, random point 2 groups, often organizes 10, is divided into normal saline group (Model group), NADPH (7.5mg/kg) dosage group; NADPH is injected in vivo by tail vein at once when myocardial ischemia-reperfusion.
(2) experimentation
Rat is set up at body Model of Myocardial Ischemia-Reperfusion Injury
The intraperitoneal injection of male adult SD rats chloral hydrate is anaesthetized, and inserts the conduit being full of heparin in right internal jugular vein and internal carotid artery, for intravenously administrable, detects arterial blood gas analysis or monitoring arteriotony.Tracheotomy also inserts endotracheal tube, connects the capable positive end expiratory pressure of ALC-V9 animal respirator, fraction of inspired oxygen 33%, regulates respiratory frequency or tidal volume, maintains pH7.35 ~ 7.45, PaCO225 ~ 40mmHg, PaO290 ~ 150mmHg.Intelligent constant-temperature controller is adopted to maintain rat temperature 36 ° of C ~ 37 DEG C.In the capable left breast otomy of the 5th intercostal, open pericardium, 6-0 non-traumatic sewing thread is at left auricle lower edge ligation ramus descendens anterior arteriae coronariae sinistrae (Left AnteriorDescending Coronary Artery, LAD), suture end penetrates self-control snare pipe, balance 30min.With mosquito forceps clamping ring sleeve pipe to block the confession of LAD blood, if visceral pericardium cyanosis is pale, electrocardio illustrates transient arrhythmia, and the ST section back of a bow is upwards raised and represented ischemia model success; Unclamp snare pipe and carry out Reperfu-sion, visible visceral pericardium is again congested proves Reperfu-sion success.Reperfu-sion 2h detects myocardial infarct size.
(3) experimental technique
The mensuration of myocardial infarct size: during myocardial ischemia in rats 30min Reperfu-sion 2h, again block LAD, internal jugular vein injection 5%EB 1ml makes left ventricle (LeftVentricle, LV) normal region indigo plant dye, rapid taking-up heart is separated LV, and use rat heart food slicer is cross-section is divided into the piece of tissue (see Fig. 1-1-3) that 5 ~ 6 pieces of 2mm are thick.By the normal structure of dye blue in LV and undyed LV ischemic region separate tissue.Adopt TTC staining, cardiac muscular tissue is put into 0.5%TTC, and 37 DEG C of water-bath 15min, 10% formaldehyde spends the night with fixing organization.LV is divided into normally under anatomic microscope, the non-infarcted region of ischemia is (red, risk zone) and (dye is for canescence in ischemic infarction district, infarct zone) 3 parts, and ischemic infarction district myocardial mass of weighing respectively accounts for the percentage ratio of ischemic region myocardial mass, the percentage ratio that myocardial infarct size accounts for ischemic region myocardial mass with infarcted region myocardial mass represents.
(4) experimental result
Fig. 5 is that therapeutic gives the impact of NADPH on myocardial ischemia infarction.Fig. 5 a is TTC coloration result; Fig. 5 b is the percentage ratio comparison diagram that ischemic infarction district myocardial mass accounts for ischemic region myocardial mass.TTC coloration result shows: compared with vehicle group (blank), reduce rat myocardial infarction model scope (p<0.05) significantly after Reperfu-sion 0h tail vein gives NADPH.Prompting NADPH reduces treating myocardial ischemia damage.Blue portion represents non-ischemic region, and RED sector represents ischemic region, and white portion represents infarcted region.Wherein
*represent p<0.05.
Obviously, above-described embodiment is only for clearly example being described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And thus the apparent change of extending out or variation be still among the protection domain of the invention.
Claims (9)
- The application of 1.NADPH in preparation treatment or prevention cardiovascular and cerebrovascular diseases medicament.
- 2. application according to claim 1, is characterized in that, described cardiovascular and cerebrovascular disease refers to the one in thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis.
- 3. application according to claim 2, it is characterized in that, described thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis refer to the thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, the atherosclerosis that are caused by vascular endothelial cell damage.
- 4. according to the application described in Claims 2 or 3, it is characterized in that, described thrombotic disease refers to the one in arterial thrombus, phlebothrombosis.
- 5.NADPH is in preparation protection vascular endothelial cell, the application safeguarded in the medicine of the normal permeability of blood vessel.
- 6., according to described application arbitrary in claim 1-5, it is characterized in that, described medicine comprises the NADPH of pharmaceutical effective amount and pharmaceutically acceptable carrier.
- 7. the medicine being used for the treatment of or preventing cardiovascular and cerebrovascular disease, it is characterized in that, described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.
- 8. the medicine being used for the treatment of or preventing any one disease in thrombotic disease, hypertension, coronary heart disease, sclerosis of blood vessels, atherosclerosis, it is characterized in that, described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.
- 9. one kind for the protection of vascular endothelial cell, the medicine safeguarding the normal permeability of blood vessel; it is characterized in that; described medicine take NADPH as active component, add in NADPH customary adjuvant conveniently technique make acceptable mixture, capsule, tablet, medicine film, spray clinically.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510086547.2A CN104840478A (en) | 2015-02-17 | 2015-02-17 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
| PCT/CN2015/095392 WO2016131321A1 (en) | 2015-02-17 | 2015-11-24 | Use of nadph in preparing medicines for treatment of cardiovascular and cerebrovascular diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510086547.2A CN104840478A (en) | 2015-02-17 | 2015-02-17 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104840478A true CN104840478A (en) | 2015-08-19 |
Family
ID=53840727
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510086547.2A Pending CN104840478A (en) | 2015-02-17 | 2015-02-17 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN104840478A (en) |
| WO (1) | WO2016131321A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105250326A (en) * | 2015-10-22 | 2016-01-20 | 苏州人本药业有限公司 | Application of NADPH (triphosphopyridine nucleotide) in preparation of antiplatelet aggregation drugs |
| WO2016131321A1 (en) * | 2015-02-17 | 2016-08-25 | 苏州人本药业有限公司 | Use of nadph in preparing medicines for treatment of cardiovascular and cerebrovascular diseases |
| CN105998048A (en) * | 2016-05-13 | 2016-10-12 | 重庆纳德福实业集团股份有限公司 | Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103340890A (en) * | 2013-06-08 | 2013-10-09 | 苏州人本药业有限公司 | Application of NADPH in preparing medicaments for preventing and treating ischemic cerebral stroke |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5945452A (en) * | 1993-06-11 | 1999-08-31 | The Board Of Trustees Of The Leland Stanford Junior University | Treatment of vascular degenerative diseases by modulation of endogenous nitric oxide production or activity |
| US5668114A (en) * | 1996-05-08 | 1997-09-16 | Birkmayer Pharmaceuticals | NADH and NADPH pharmaceuticals for treating hypertension |
| CN104840478A (en) * | 2015-02-17 | 2015-08-19 | 苏州人本药业有限公司 | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases |
-
2015
- 2015-02-17 CN CN201510086547.2A patent/CN104840478A/en active Pending
- 2015-11-24 WO PCT/CN2015/095392 patent/WO2016131321A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103340890A (en) * | 2013-06-08 | 2013-10-09 | 苏州人本药业有限公司 | Application of NADPH in preparing medicaments for preventing and treating ischemic cerebral stroke |
Non-Patent Citations (3)
| Title |
|---|
| 王萌等: "氧化应激与糖尿病周围神经病变", 《临床荟萃》 * |
| 白玉婷等: "氧化应激与心血管疾病关系的研究进展", 《医学综述》 * |
| 邵凤民: "血管内皮细胞与血栓形成的关系", 《河南医学研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016131321A1 (en) * | 2015-02-17 | 2016-08-25 | 苏州人本药业有限公司 | Use of nadph in preparing medicines for treatment of cardiovascular and cerebrovascular diseases |
| CN105250326A (en) * | 2015-10-22 | 2016-01-20 | 苏州人本药业有限公司 | Application of NADPH (triphosphopyridine nucleotide) in preparation of antiplatelet aggregation drugs |
| CN105998048A (en) * | 2016-05-13 | 2016-10-12 | 重庆纳德福实业集团股份有限公司 | Pharmaceutical composition for treating ischemic cerebral apoplexy and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2016131321A1 (en) | 2016-08-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2019504819A (en) | Use of kaurane compounds in the manufacture of a medicament for the treatment of cardiac hypertrophy and pulmonary hypertension | |
| CN104840478A (en) | Application of NADPH in preparation of drugs used for treating cardio-cerebrovascular diseases | |
| CN101347422B (en) | Uses of salvianolic acid A in preventing and/or treating diabetes and complication | |
| Burman et al. | Evaluation of toxicity and antitumor activity of cycloviolacin O2 in mice | |
| CN103948619B (en) | Application of vaccarin for resisting oxidation and high-glucose damage | |
| US20090036527A1 (en) | Therapeutic Application Of Leonurine In Treating Cardiomyopathy | |
| CN103608026B (en) | Peptide combinations and for treating injury of lung, asthma, anaphylaxis, angioedema, the penetrating syndrome of Systemic Vascular and the method for nasal obstruction | |
| CN104840479A (en) | Application of NADPH in preparation of drugs used for treating heart diseases | |
| CN107158008B (en) | A kind of pharmaceutical composition for treating myocardial infarction | |
| CN106474479B (en) | Angiotensin receptor antagonist and the compound of Creatine Phosphate Sodium and application thereof | |
| CN101549014A (en) | Heart-protecting musk oral preparation and preparation method thereof | |
| CN106214680B (en) | A kind of compound of angiotensin receptor antagonist and Levosimendan and application thereof | |
| CN103784934B (en) | A kind of pharmaceutical composition and application thereof preventing and treating acute myocardial infarction | |
| CN101070338A (en) | Tanshinone IIA potassium sulfonate used for preparing medicine for preventing and treating cerebral ischemia and cerebral anoxia and myocardial ischemia and myocardio anoxia | |
| CN102824339B (en) | Application of sodium butyrate in preparation of hypoxic pulmonary hypertension control medicines | |
| CN106421746B (en) | The application of the salvage drug of cardiac arrest caused by [Pyr1] Apelin 13 is poisoned as long-acting local anesthetics of amide derivatives | |
| CN106902131A (en) | Applications of the NADPH in medicine of the treatment myocardial hypertrophy with heart failure is prepared | |
| CN107929262A (en) | Ethylenediamine cationized albumin anti-tumor nano grain and its preparation method and application | |
| CN112755015A (en) | Application of PT2385 in preparation of medicine for preventing and treating pulmonary hypertension | |
| CN103203009B (en) | New application of partial metabolite of pentapeptide in preparation of myocardial ischemia resistant product | |
| CN110051664A (en) | A kind of application of ginkgo lactone composition in drug of the preparation for acute coronary syndrome | |
| RU2437692C2 (en) | Method of treating solid malignant growths and their metastases | |
| CN107569476A (en) | Pharmaceutical composition containing Acetylshikonin and its application in pulmonary hypertension medicine | |
| CN104189288B (en) | A kind of Chinese medicine composition for treating myocardial infarction and its application | |
| CN110075131A (en) | Application of the zika virus attenuated strain in treatment glioma |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150819 |