CN101612157A - Fasudil is induced the purposes of the brain endogenous neural stem cell regeneration of growing up - Google Patents
Fasudil is induced the purposes of the brain endogenous neural stem cell regeneration of growing up Download PDFInfo
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
- A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
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- A—HUMAN NECESSITIES
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- A61P25/00—Drugs for disorders of the nervous system
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- A61P25/16—Anti-Parkinson drugs
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
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Abstract
The pharmaceutical composition that the invention discloses fasudil and comprise fasudil is used to induce the purposes of brain endogenous neural stem cell regeneration of growing up; especially, the invention discloses the purposes that fasudil is used for the neuroprotective damage disease relevant with death with treatment and neuronal damage.
Description
Technical field
The present invention relates to the purposes that fasudil is used to induce adult brain endogenous neural stem cell regeneration, especially, relate to the purposes that fasudil is used for the neuroprotective damage disease relevant with death with treatment and neuronal damage.
Background technology
Fasudil is the isoquinolin sulphone amide derivative, and chemical name is six hydrogen-1-(5-isoquinolinesulfonylcompounds)-1H-1, the high piperazine of 4-diazepine or 1-(5-isoquinolinesulfonylcompounds), and English name is Fasudil, molecular formula is C
14H
17N
3O
2S.Its customary salt has hydrochlorate, nitrate, sulfate, mesylate, and fasudil hydrochloride is the most normally used a kind of salt.2002, fasudil went on the market in China, and indication is the improvement of the ischemic cerebrovascular symptom that causes such as cerebral vasospasm after subarachnoid hemorrhage.The hemorrhage back of angiorrhexis in brain blood flow is gone into subarachnoid space, is called subarachnoid hemorrhage, and it is a type of hemorrhagic apoplexy, divides two kinds of constitutional and Secondary cases.Headache, vomiting and stiffness of the neck are the cardinal symptoms of subarachnoid hemorrhage.Cerebral vasospasm is meant the abnormal contraction state of tremulous pulse in a period of time, is common in the subarachnoid hemorrhage patient of rupture of intracranial aneurysm.Cerebral vasospasm is disabling of subarachnoid hemorrhage and main causes of death.Shown that fasudil is a kind of novel, vasodilator (its for a kind of kinases inhibitor, i.e. intracellular calcium antagonist) efficiently, can effectively alleviate cerebral vasospasm, improved the cerebral vasospasm that causes behind the subarachnoid hemorrhage.The effectiveness and the safety of the cerebral vasospasm that causes behind existing many at present clinical reports confirmation Fasudic hydrochloride treatment subarachnoid hemorrhages.
Fasudic hydrochloride pharmacokinetics and metabolic activity product have been studied in existing in vivo test.Fasudic hydrochloride enters to change into very soon in the body has identical active metabolite-HA 1100 (hydroxyfasudil), its concentration can reach 86% of female medicine, and have the long half-life (0.78 hour Fasudic hydrochloride half-life, and 4.66 hours HA 1100 half-life).12 hours peak-to-valley values of HA 1100 are 0.077 μ M, and have the biological activity identical with Fasudic hydrochloride, can (0.025-1.6 μ M) suppress the Rho kinases in the valid density scope.
Nervous system disease such as cerebral ischemia, brain injury, Alzheimer, parkinson disease, retinal diseases can cause neuronic damage and death in its generation and evolution.For this group disease, people are seeking effective medicine always energetically.Past is over 20 years; people study and have observed a large amount of neuroprotectives; attempt to prevent and to treat the generation and the development of these nervous system disease; comprise calcium-channel antagonists, free radical scavenger, glutamate antagonist, cell membrane stability agent and neurotrophic factor etc.; but owing to multiple factor while or priority behind the brain injury have participated in neuronic damage with dead; the medicine that acts on single target spot merely is difficult to obtain gratifying curative effect, and the clinical therapeutic efficacy of above-mentioned neuroprotective is unsatisfactory.
Regeneration and Repair problem behind the nervous system injury is the emphasis that the neuroscience field is paid close attention to always.Along with neural stem cell research deepens continuously, people attempt utilizing the Neural Stem Cells Repairing injured neurons to become one of focus of neuroscience area research in recent years.Neural stem cell plays a significant role in repairing damaged nerve tissue, is the effective ways of repairing and replace damaged tissues, can rebuild part loop and function.At present, studying more widely, graft mainly still is embryonic stem cell and neural stem cell.But problems such as, ethics obstacle limited because of originating and immunologic rejection, the practicality of these cells is subjected to very big restriction.Recently studies show that mesenchymal stem cells MSCs has the ability that is divided into neuron and neurogliocyte.But bone marrow-derived cells such as mesenchymal stem cells MSCs have higher risk from viral infection, and along with its source individual its cell quantity of age growth and propagation, differentiation capability also significantly descend, difficulty satisfies clinical demand.
1992, Reynolds etc. at first isolated neural stem cell from the adult rats striatum, thereby had broken through the non-renewable viewpoint of adult tissue.This experiment proves that fully the newborn neuron of the neural stem cell generation of growing up is not only identical with mature neuron on form, and synapse stimulated the action potential reaction is arranged, and can be incorporated in the neural loop effectively, thereby,, we have indicated a direction for utilizing the neural stem cell of growing up to carry out the brain repairing and treating for the clinical practice neural stem cell is laid a good foundation.But up to the present, this research field does not still find and a kind ofly can act on adult neural stem cell effectively in vivo, induces its regeneration, thereby repairs the injured nerve cell to reach the medicine of treatment nervous system disease.Therefore, seek a kind of can be free by blood brain barrier, metastable, can to induce the endogenous regenerated material of stem cell of cranial nerve of growing up be the direction that people strive to find always.
Summary of the invention
The inventor finds that unexpectedly fasudil also has the neuroprotective function and has the stem cell of cranial nerve differentiation that promotes to grow up, induces the regenerated function of stem cell of cranial nerve of growing up, thereby has finished the present invention through research in earnest for a long time.
Therefore, in one aspect, the invention provides the purposes of fasudil neuroprotective function.
In one aspect, the invention provides fasudil and induce the regenerated purposes of stem cell of cranial nerve of growing up, promptly induce the purposes of the brain endogenous neural stem cell of growing up.
In yet another aspect, the invention provides the purposes of the fasudil treatment disease relevant with death, promptly treat the purposes of nervous system disease with neuronal damage.
In one aspect, the invention provides fasudil is used for inducing the adult regenerated medicine of stem cell of cranial nerve in preparation purposes.Especially, the invention provides fasudil is preparing as the purposes in the medicine of neuroprotective.
In yet another aspect, the invention provides fasudil and be used for the treatment of purposes in the medicine of nervous system disease in preparation.
Aspect going back one, the invention provides a kind of pharmaceutical composition for the treatment of nervous system disease, it comprises the fasudil or the Fasudic hydrochloride for the treatment of effective dose.
Fasudil of the present invention, comprise pharmaceutically acceptable salt forms such as its hydrochlorate, nitrate, sulfate, phosphate, hydrobromate, mesylate, citrate, it can be arbitrary form, comprise pharmaceutically acceptable forms such as crystal form (comprise have a water of crystallization and do not have water of crystallization), hydrate forms, preferred fasudil hydrochloride.These salt and different forms can be according to method preparations well known in the art.
To be fasudil be used for the amount that neuroprotective or treatment nervous system disease institute may produce the effect of medically approving to effective dose of the present invention.The inventor is through repeatedly discovering, is suitable for inducing the stem cell of cranial nerve regeneration of growing up, and the effective dose that promptly is suitable for treating the fasudil of nervous system disease is 0.5-8mg/kg, preferred 1-6mg/kg, more preferably 3-5mg/kg, most preferably 5mg/kg.For the various salt of above-mentioned fasudil and comprise the consumption of the pharmaceutical composition of fasudil, be the effective dose of corresponding salt with the amount of the corresponding fasudil salt that is equivalent to provide above-mentioned fasudil effective dose.
The disease relevant with death with neuronal damage of the present invention, promptly described nervous system disease includes, but are not limited to cerebral ischemia, brain injury, Alzheimer, parkinson disease, dementia, retinal diseases etc.
Can be according to routine techniques known in the art, fasudil of the present invention or the pharmaceutical composition that comprises the fasudil of effective dose are made any dosage form that is suitable for administration, include but not limited to intestinal canal administration preparation or non-intestinal drug delivery agent, for example tablet, capsule, oral liquid, injection, injectable powder etc., optimizing injection, more preferably intravenous injection.
The route of administration of preparation of the present invention can be any route of administration known in the art, preferred lumbar injection.
Preparation of the present invention can administration every day 3 times, and each dosage is the effective dose that fasudil can be treated described disease, the promptly above-mentioned effective dose that provides.
Be to be understood that, for mentioning among the present invention but the method that does not have to describe in detail, step, device, instrument, material etc., those of ordinary skill can adopt correlation method well known in the art, step, device, instrument, material etc., and perhaps the conventional knowledge and technology according to this area obtains.
Description of drawings
Hippocampal neuron MAP-2 and the two colored graphs of HOCHEST that Fig. 1 is commissioned to train foster for the display body exogenesis.Redness is the MAP-2 neuron, and blueness is the HOCHEST nuclear staining, in order to check this institute with neuronic form and purity.
Fig. 2 is for showing mice at normal oxygen and cerebral anoxia (8% oxygen) after 6 hours, and reoxygenation is Hippocampus different parts neuron colored graph after 72 hours.CA1, CA2 and CA3 are respectively Hippocampus CA1, CA2 and CA3 district.
Fig. 3 measures the figure of the proteic expression of the kinase whose hypotype RockII of Rho for showing mice immunoblotting after cerebral anoxia (8% oxygen) different time brain tissue homogenate.
Fig. 4 is for showing the figure of mice in the expression of the different kinase whose hypotype ROCK II of time brain tissue slice immunohistochemical staining Rho of cerebral anoxia, and the ROCK II that the reddish yellow arrow is green fluorescence expresses.
Fig. 5 is for showing mouse brain anoxia injecting normal saline and fasudil (mg/kg) after (8%) 6 hour, tricorn inferior segment (SVZ) relief angle (A and B) after 24 hours, the figure of anterior angle (C and D) and Hippocampus DG district (E and F) Brdu positive cell number.
Fig. 6 injects fasudil for showing the mouse brain anoxia after (8%) 6 hour, the figure of the two positive cell numbers of tricorn inferior segment (SVZ) relief angle DCX and Brdu after 24 hours.A) redness is the Brdu positive, cell, and B) the green DCX of being dyes, and C) two kinds of colors is overlapping.The overlapping cell of redying is outgrowth neural stem cell.
Fig. 7 is for showing normal saline group and the mouse brain tissue homogenate supernatant G-CSF of fasudil group and the concentration level figure of VEGF.Adopt the detection kit of import standard to measure.
Fig. 8 is the display body exogenesis is cultivated the Brdu positive cell number of back and PBS contrast for neuron figure.
Fig. 9 causes culture supernatant LDH to discharge the figure of minimizing and the minimizing of PI positive cell number for neuronal death after showing fasudil inhibition anoxia.
The specific embodiment
The present invention will do further in conjunction with following test example or embodiment and elaborate, but should be appreciated that following embodiment only is used for setting forth and explaining the present invention, and not limit the scope of the invention.
Test example 1: the external promotion stem cell of cranial nerve of fasudil hypertrophy test
1. test cell and preparation thereof
The pregnant Mus of pregnant 17-18 days KM is provided by Shanghai Slac Experimental Animal Co., Ltd..Take off the neck method and put to death pregnant Mus, 75% alcohol disinfecting skin.Under aseptic condition, strip out the tire Mus, place the DMEM culture fluid of ice pre-cooling.The careful tire murine brain that separates divests meninges and blood vessel, removes cerebellum, brain stem and Basal ganglia, takes out hippocampal tissue, places the DMEM of ice pre-cooling to blow and beat gently, makes it form single-cell suspension liquid.The collecting cell suspension, under 4oC, centrifugal 5 minutes with 1000 rev/mins.Abandoning supernatant suspends again with freshly prepared neuronal cultured solution.Counting cells, the density of cell in the calculating cell suspension.With 2 * 10
5/ cm
2Be planted on the slide of poly-D-lysine bag quilt, under 37 ℃, 5%CO
2Cultivate in the incubator, change 1/2 culture fluid after 7 days.Carry out the neuronal specificity labelling after 14 days: microtubule-associated protein-2 (MAP-2) dyeing is identified, simultaneously with fluorescent dye HOCHEST33342 labeled cell nuclear.Observed result (see figure 1) under the fluorescence microscope.The MAP-2 positive cell is red, prompts for neuron, and blueness is a nucleus.Neuronal cell after the evaluation lacks sugared anoxia-induced apoptosis (oxygen-glucose deprivation, OGD) model for setting up.
2. lack sugared hypoxic cell damage model
Sugarless type DMEM culture fluid is with preceding feeding 5%CO
2, 10%H
2, 85%N
2The saturated 15min of mist, the oxygen in the culture fluid of place to go, preparation lacks sugared anoxia neuronal cultured solution.Get and cultivated about 10 days, well-grown neuron discards culture fluid, with PBS liquid flushing 3 times, adds Sugarless type DMEM culture fluid.Culture plate is placed in the anaerobism incubator, in case, feed 5%CO
2, 10%H
2, 85%N
2Mist keeps 37 ℃ with the incubator sealing, and makes the interior oxygen concentration of case be lower than 1%.After certain anoxia time (for example 6 hours), culture plate is taken out, absorb the sugar-free culture fluid, add former neuronal cultured solution respectively, be positioned over 37 ℃, 5%CO
2Cultivate with continuing in the incubator of 95% air.
3. administration
Subsequently, administration lacks sugared anoxia neuron fasudil injection (female concentration is 30mg/2ml=15mg/ml).Experimental concentration is set at height, and low three concentration of neutralization are respectively 1 μ g/ml, 10 μ g/ml and 100 μ g/ml.Added for three times on the same day, the 2nd day and the 3rd natural gift respectively.The 5th day collecting cell and supernatant are done further detection and are used.
4. controlled trial:
After experimental model is determined, adopt the PBS that adds equal volume to replace the fasudil injection, under identical experiment condition as the contrast of fasudil.Cultured cells is as the baseline of hypoxic cell under the normal oxygen condition.
5. neuron BrdU detects
Add fasudil after the 3rd day, added Brdu (1ug/ml) respectively at the 3rd and the 4th day.The 5th day collecting cell PBS rinsing cell 10min * 2 times, 4% paraformaldehyde room temperature is 20min fixedly, 1 * PBS rinsing cell 10min * 2 times, 3% sheep blood serum room temperature sealing 15min.BrdU one anti-(1: 200) is diluted in and contains among the 1% sheep blood serum PBS, 4 ℃, spends the night.Goat-anti mice CY3 labelling two anti-(1: 100) is diluted among the PBS room temperature lucifuge 1 hour.1 * PBS rinsing cell 5min * 3 times, 50% glycerol mounting, observed result under the fluorescence microscope.
6. result and analysis
Fig. 8 shows comparison PBS group, adds fasudil group neuron and forms comparatively tangible neural ball.The male hypertrophy neuronal cell of Brdu is obviously more than the PBS matched group.The result clearly illustrates that fasudil can be in external promotion stem cell of cranial nerve hypertrophy.Test of many times obtains identical result.
Test example 2: promote the test of stem cell of cranial nerve hypertrophy in the fasudil body
1. experimental animal
The KM mice is provided by Shanghai Slac Experimental Animal Co., Ltd..Animal is given and the raising of cleaning level after arriving laboratory, and free diet and drinking-water were rested 5-7 days.
2. anoxia in mice model
32 mices are divided into 4 groups, 8 every group, in oxygen depletion system, give to distinguish in 8% oxygen+92% nitrogen mixture body anoxia 2 hours, 4 hours, 6 hours, 8 hours.In the anoxia case, place the anoxia instrument, oxygen concentration in the detection of dynamic anoxia case, the controllability of assurance anoxia in mice degree.Casing can hold 30-40 mice, and 32 mices are once put into the comparability that can guarantee anoxia condition.-80 ℃ of preservations behind the brain liquid nitrogen flash freezer are got in reoxygenation normal saline perfusion after 72 hours, and behind the brain tissue slice, hippocampal neuron and ROCK II dye when treating immunity printing and dyeing and SABC.
3. administration
Fasudil injection, female concentration are 30mg/2ml=15mg/ml.According to the result of experiment in vitro, in vivo test concentration is set at mice 6 hours pneumoretroperitoneums of anoxia injection Fasudic hydrochloride (5mg/kg) and Brdu (50mg/kg) in oxygen depletion system, and duplicate injection is once again after 24 hours; Matched group injection equal volume normal saline and Brdu.8 Mus of fasudil group, and 7 Mus of normal saline matched group.-80 ℃ of preservations behind the brain liquid nitrogen flash freezer are got in reoxygenation normal saline perfusion after 24 hours, behind the brain tissue slice, carry out Brdu and DCX and dye when treating immunity printing and dyeing and SABC.
4. controlled trial:
When experimentizing model, adopt the physiologic saline for substitute fasudil injection of injection equal volume, under identical experiment condition as the contrast of fasudil.Setting 8 of normal oxygen animals in addition carries out in contrast under identical condition.
5. the immunity printing and dyeing are measured
After the discontinuous gel electrophoresis of 10%SDS-PAGE separates, change film 15-30min on pvdf membrane, the nitrocellulose membrane after the transfer places the Ponceaux dyeing liquor to develop the color, and after labelled protein Marker and the band position, transfering buffering liquid is washed till decolouring.The sealing nitrocellulose membrane places membrane closure liquid, slowly shakes room temperature sealing 1h on the shaking table.Anti-ROCK II antibody spends the night with 5% calf serum dilution (1: 500) 4 ℃.Adopt two dilutions in anti-1: 1000, incubated at room 1h.With chemoluminescence method colour developing, X-ray film developing.The collection of illustrative plates of immunoblotting is taken pictures by digital camera, through Image-pro Plus 5.0 image processing softwares, is confidential reference items with β-actin.
6. SABC is measured
Mice broken end tailing edge midline incision scalp is also cut off skull along sagittal suture, takes out brain, immerses 4 ℃ of dehydrations in 20% sucrose, places 4 ℃ of dehydrations of 30% sucrose to tissue to sink after sinking to the bottom again.Blot the sucrose solution on cerebral tissue surface with filter paper,, make the freezing serial section in crown position, deposit standby according to sequencing with the thickness of 8 μ m with OCT embedding in liquid nitrogen.Immunofluorescence dyeing is with sealing 10min under the 5% sheep blood serum room temperature, one anti-4 ℃ hatch 24 hours after, two anti-room temperature lucifuge effects of fluorochrome 488 labellings 1 hour, behind 1 * PBS washing 5min * 3 time, 50% glycerol mounting is observed, is taken pictures under the fluorescence microscope.
7. result and analysis
The result shows that the anoxia model that we set up can cause neuronic losing (referring to Fig. 2) in Hippocampus zones of different, particularly CA1 and the CA2 district.Method with immunity printing and dyeing and SABC proves that the expression of cerebral anoxia ROCK II after 6 hours increases, and provides according to (Fig. 3 and Fig. 4) for using the Rho inhibitors of kinases.The injection fasudil can obviously increase brain SVZ district's anterior angle and relief angle position Brdu positive cells regeneration (Fig. 5) after the result showed anoxia.Discover that further the male proliferative cell of these Brdu expresses neural stem cell sign DCX (Fig. 6) simultaneously, illustrate that the inductive SVZ of fasudil district proliferative cell is a neural stem cell.
Test example 3: fasudil is to neuronic protective effect
1. test cell
With 1. test cells of test example 1 and the method preparation test cell described in the preparation thereof.
2. lack sugared anoxia-induced apoptosis model
Lack the described method preparation of sugared anoxia-induced apoptosis model with 2. of test example 1 and lack sugared anoxia-induced apoptosis model.
3. administration
Compare preliminary experiment according to fasudil variable concentrations in the test example 1, we have selected intermediary concentration in this experiment, i.e. 10 μ g/ml.All the other are identical in the example 1 with test.
4. controlled trial
After test model is determined, adopt add equal volume PBS under identical experiment condition as the contrast of fasudil.
5. cell LDH release rate assay
After determining that giving the neuron different time handles, the situation of neuronal damage in the time of different complex sugar reoxygenations is by measuring the degree that lactic acid dehydrogenase (LDH) detects cell injury.Intracellular LDH will be discharged in the cell culture fluid after neuron sustains damage.Thereby detect the interior LDH value of damage back cell conditioned medium liquid, reacted the cell damage situations.Normal cell is given to freeze repeatedly molten, the LDH amount for discharging behind whole cell deaths in its supernatant.The cell conditioned medium liquid LDH value that is detected is compared with the dead fully total LDH value that discharges of cell, obtained cell LDH release rate.LDH measures and operates according to the test kit description.Wavelength 490nm detects the OD value in every hole on microplate reader.Cell LDH release rate is represented with total OD value * 100% that discharges of test set OD value/cell.
6.PI the dyeing flow cytometer detects
(propidine iodide PI) is a kind of nucleic acid dye to propidium iodide, and it can not see through complete cell membrane, but at the cell and the dead cell of apoptosis middle and advanced stage, and PI can permeate through cell membranes and made that nucleus is red to be dyed.Therefore, utilize flow cytometer to detect the PI positive cell and represent dead neuronal cell number.
7. result and analysis
Fig. 9 shows that detecting the processing of discovery fasudil with the LDH test kit can reduce LDH concentration in the neuronal cell culture supernatant (p＜0.05), handles with flow cytometer detection discovery fasudil and can reduce PI cell number (p＜0.01).We adopt diverse ways (LDH discharge and flow cytometer detection) to confirm that all fasudil can protect neuronic damage after the anoxia.
Test example 4: the mechanism of action of fasudil induced nerve stem cells
1. experimental animal
With 1. test cells of test example 2 and the method preparation test cell described in the preparation thereof.
2. anoxia in mice model
16 mices give to distinguish in 8% oxygen+92% nitrogen mixture body anoxia 6 hours respectively in oxygen depletion system.In the anoxia case, place the anoxia instrument, oxygen concentration in the detection of dynamic anoxia case, the controllability of assurance anoxia in mice degree.Fasudil group and normal saline group are respectively 8.
3. administration
Fasudil injection (female concentration is 30mg/2ml=15mg/ml).Result according to experiment in vitro, in vivo test concentration is set at mice 6 hours pneumoretroperitoneums of anoxia in oxygen depletion system and injects Fasudic hydrochloride (5mg/kg) and Brdu (50mg/kg), after 24 hours again duplicate injection once, matched group injection equal volume normal saline and Brdu.8 Mus of fasudil group, 8 Mus of normal saline matched group.-80 ℃ of preservations behind the brain liquid nitrogen flash freezer are got in reoxygenation normal saline perfusion after 24 hours, carry out brain tissue homogenate's supernatant cytokine assay.
4. controlled trial:
When experimentizing model, the normal saline that adopts the injection equal volume under identical experiment condition as the contrast of fasudil.
5. cytokine assay
For further furtheing investigate the cellular elements mechanism that fasudil is induced the endogenous neural stem cell, the test kit that we adopt the merchant to sell detects G-CSF and VEGF concentration in brain tissue homogenate's supernatant.Detect step with reference to description.
6. result and analysis
The fasudil processing had increased the concentration of G-CSF in the cerebral tissue (p＜0.01) significantly after result of the test showed anoxia, and had reduced the level (p＜0.05) (referring to Fig. 7) of VEGF.A large amount of experiments have proved that G-CSF can induce the endogenous neural stem cell to mobilize, and show that fasudil induces the endogenous neural stem cell to increase with G-CSF and VEGF reduces relevant.
Claims (9)
1. fasudil is used for inducing the purposes of the adult regenerated medicine of stem cell of cranial nerve in preparation.
2. the purposes of claim 1, wherein said adult stem cell of cranial nerve is the brain endogenous neural stem cell.
3. fasudil is the purposes of neuroprotective at the medicine that preparation is used for the neuroprotective function.
Fasudil the treatment nervous system disease medicine in purposes.
5. each purposes among the claim 1-4, wherein said fasudil is selected from fasudil hydrochloride, nitrate, sulfate, phosphate, hydrobromate, mesylate or citrate.
6. the purposes of claim 5, wherein said fasudil is a fasudil hydrochloride.
7. claim 4 or 6 purposes, wherein said nervous system disease is selected from cerebral ischemia, brain injury, Alzheimer, parkinson disease, dementia or retinal diseases.
8. the purposes of claim 5, wherein said nervous system disease is selected from cerebral ischemia, brain injury, Alzheimer, parkinson disease, dementia or retinal diseases.
9. the purposes of claim 7, wherein said nervous system disease is selected from cerebral ischemia or brain injury.
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| CN200810126351A CN101612157A (en) | 2008-06-26 | 2008-06-26 | Fasudil is induced the purposes of the brain endogenous neural stem cell regeneration of growing up |
| CN2009801229640A CN102026643B (en) | 2008-06-26 | 2009-05-11 | The use and method of the compound of fasudil and the pharmaceutical composition thereof |
| PCT/CN2009/000509 WO2009155777A1 (en) | 2008-06-26 | 2009-05-11 | The use and method of the compound of fasudil and the pharmaceutical composition thereof |
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| CN104557897A (en) * | 2014-04-10 | 2015-04-29 | 中山大学 | Fasudil-lipoic acid dyad and application thereof |
| CN113440649A (en) * | 2021-05-31 | 2021-09-28 | 温州医科大学 | Spinning membrane scaffold for directionally inducing axon regeneration |
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| CN102973571A (en) * | 2012-12-12 | 2013-03-20 | 天津红日药业股份有限公司 | New application of fasudil |
| CN104983704A (en) * | 2015-08-13 | 2015-10-21 | 青岛蓝盛洋医药生物科技有限责任公司 | Medicine fasudil hydrochloride composition tablet capable of relieving cerebral angiospasm |
| CN105055332A (en) * | 2015-09-10 | 2015-11-18 | 青岛蓝盛洋医药生物科技有限责任公司 | Hydroxyfasudil composition dry suspension for treating ischemic cerebrovascular diseases |
| CN115103672A (en) | 2020-01-09 | 2022-09-23 | 悟而喜制药公司 | Methods of treating wandering associated with cortical dementia |
| EP4125879A4 (en) | 2020-03-25 | 2024-04-03 | Woolsey Pharmaceuticals Inc | Methods of treating proteinopathy- associated wandering |
| MX2023004523A (en) | 2020-10-22 | 2023-05-08 | Woolsey Pharmaceuticals Inc | Methods of treating 4-repeat tauopathies. |
| WO2023097151A1 (en) | 2021-11-29 | 2023-06-01 | Woolsey Pharmaceuticals, Inc. | Methods of treating agitation and other dementia-associated behavioral symptoms |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2005117896A1 (en) * | 2004-06-03 | 2005-12-15 | Schering Aktiengesellschaft | Formulations containing fasudil, a matrix and an envelope |
| CN1729985A (en) * | 2005-08-02 | 2006-02-08 | 吴良信 | Fasudil hydrochloride injection and its preparation process |
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2008
- 2008-06-26 CN CN200810126351A patent/CN101612157A/en active Pending
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104557897A (en) * | 2014-04-10 | 2015-04-29 | 中山大学 | Fasudil-lipoic acid dyad and application thereof |
| CN104557897B (en) * | 2014-04-10 | 2017-10-27 | 中山大学 | The synthesis and its application of Fasudil lipoic acid dyad |
| CN113440649A (en) * | 2021-05-31 | 2021-09-28 | 温州医科大学 | Spinning membrane scaffold for directionally inducing axon regeneration |
Also Published As
| Publication number | Publication date |
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| CN102026643A (en) | 2011-04-20 |
| CN102026643B (en) | 2013-05-01 |
| WO2009155777A1 (en) | 2009-12-30 |
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