CN102199123B

CN102199123B - New crystal form and pharmaceutical application of fasudil

Info

Publication number: CN102199123B
Application number: CN201010130620.9A
Authority: CN
Inventors: 姚小青
Original assignee: Tianjin Chase Sun Pharmaceutical Co Ltd
Current assignee: Tianjin Chase Sun Pharmaceutical Co Ltd
Priority date: 2010-03-24
Filing date: 2010-03-24
Publication date: 2014-09-03
Anticipated expiration: 2030-03-24
Also published as: CN102199123A

Abstract

The invention relates to a new crystal form and pharmaceutical application of fasudil, particularly to a crystal form of fasudil hydrochloride. The new crystal form of fasudil hydrochloride is characterized by having the following spectroscopic characteristics that the crystal phase of a sample is analyzed by a Rigaku D/max-2500 X-ray powder diffraction (XRD) instrument with a Cu Kalpha target at a tube voltage of 40 KV and a tube current of 100 mA, and the X-ray powder diffraction of the sample has the following characteristic peaks: peaks exist at angles 2theta which are about 6.780 DEG, 14.700 DEG, 16.620 DEG, 17.480 DEG, 20.440 DEG, 20.740 DEG and 24.52 DEG.

Description

A kind of crystal formation of fasudil and medicinal application thereof

Technical field

The present invention relates to a kind of crystal formation of chemicals Fasudil Hydrochloride.The invention still further relates to and use the ischemic cerebrovascular that this crystal formation improves and prevention is caused by many reasons, such as after the Delayed onset cerebro-vascular diseases (DINDS) causing after cerebral infarction, vertebro-basilar artery insufficiency, subarachnoid hemorrhage, cerebral surgery operation and the cerebral vasospasm that causes of PTCA or and STENTS, transient ischemic attack (TIA), hematencephalon decubation, Neurology Department cerebral infarction etc. cerebral ischemia relative disease.

Background technology

Fasudil [six hydrogen-1-(5-isoquinolinesulfonylcompounds)-homopiperazine, Fasudil] is a kind of novel isoquinoline sulphone amide derivative.Its molecular weight is 327.83, and molecular formula is C14H17N3O2SHCl:

Fasudil has therapeutic action to ischemic cerebrovascular disease, can remove cerebral vasospasm, increases cerebral blood flow (CBF), and brings into play cerebral protection, control chronic ischemic cerebral vasospasm.Fasudil Hydrochloride be in the world a unique Rho kinase inhibitor of having gone on the market (referring to Lin Xiangyu etc., the treatment research of Rho kinase inhibitor to acute ischemic cerebral apoplexy, contemporary Chinese medicinal application, 2008,2 (13) 23-4; Zhang Zuyu etc., the routine clinical observation of Treatment of Acute Cerebral Infarction with Hydrochloric Fasudil 43, Shandong medicine, 2008,48 (41) 104-104).There is the effect (referring to Chinese patent application 200910016702.8) of the adult brain endogenous neural stem cell regeneration of induction.

Fasudil has multiple preparation method, but its end product has armorphous feature conventionally, and the present invention, by research, finds a kind of new crystal formation of Fasudil Hydrochloride, the stability having had and storage characteristics and preparation industrial utilization.

Summary of the invention

The invention provides under the normal temps of storing and using and have degree of physical stability, and there is the Fasudil Hydrochloride crystal formation of peculiar property.

This invention provides a kind of crystal formation of Fasudil Hydrochloride, it is characterized in that, there is following spectrum characteristic, adopt Rigaku Rigaku D max-2500 type X powder diffraction (XRD) instrument to analyze the crystalline phase of sample, Cu K α target, tube voltage 40KV, tube current 100mA, its X-powdery diffractometry has following characteristic peak: at approximately 6.780,14.700,16.620,17.480,20.440,20.740,24.52 degree 2 θ places, have peak.

The crystal formation of Fasudil Hydrochloride of the present invention has following spectrum characteristic and physics-chem characteristic:

1.X-powdery diffractometry

Adopt Rigaku Rigaku D max-2500 type X powder diffraction (XRD) instrument to analyze the crystalline phase of sample, Cu K _αtarget, tube voltage 40KV, tube current 100m A.Its X powder diffraction has following characteristic peak: table 1

Angle (2 θ)	D-value	Intensity (Cps)	Intensity (%)
				6.780	13.0264	8726	18
10.040	8.8029	3199	7
				11.580	0.141	1525	3
12.640	0.165	1425	3
				13.480	0.259	4493	9
14.700	0.188	8701	18
				16.100	0.165	3450	7
16.620	0.188	17364	36
				17.480	0.188	8268	17
18.880	0.165	2001	4
				20.440	0.165	47857	100
20.740	0.235	8427	18
				21.480	0.188	7133	15
21.940	0.329	2350	5

23.280	0.165	978	2
				24.080	0.165	3730	8
24.520	0.212	16659	35
				25.480	0.188	5688	12
27.380	0.188	3933	8
				28.260	0.188	4188	9
29.700	0.188	2011	4
				29.980	0.165	2997	6
30.260	0.118	2955	6
				30.460	0.212	2755	6
31.240	0.188	1031	2
				32.600	0.212	2085	4
33.320	0.212	2230	5
				33.980	0.235	1558	3
34.400	0.188	3409	7
				35.760	0.188	996	2
36.920	0.235	3306	7
				39.280	0.235	2425	5
40.720	0.165	1756	4
				41.100	0.235	1642	3
41.580	0.188	1744	4
				43.860	0.235	1440	3
44.720	0.259	1113	2

2. fusing/decomposition temperature

With Mettler Toledo DSC 822e differential scanning calorimeter, measure the fusing/decomposition temperature of fasudil.5.8700mg fasudil is placed in one, with the heat-up rate heating of approximately 10 ℃/min.Fusing/decomposition temperature from fusing/decompose endotherm extrapolation start to maximum value, define.The fusing of fasudil is with decomposition, and analyzed the impact of front solids treatment.Fusing/the decomposition temperature of this crystal formation is 214.46-219.41 ℃.

3. infrared spectra

The peak position 3426.65cm of solid-state FT infrared spectral band ^-1nH stretching vibration, 1614.68cm ^-1, 1584.99cm ^-1the stretching vibration of isoquinoline 99.9 skeleton, 1336.57cm ^-1sO ₂asymmetrical stretching vibration, 1157.09cm ^-1sO ₂symmetrical stretching vibration.

This crystal formation is peak position 3426.65cm with the difference containing crystal water ^-1nH stretching vibration, contains crystal water crystal formation at 3500cm ^-there is no obvious peak.

4. ¹³c nuclear magnetic resonance spectrum

Fasudil ¹³as shown in Figure 4, by analyzing, fasudil crystal formation 110-160ppm's is the carbon resonance on isoquinoline 99.9 to the example of C nuclear magnetic spectrum.

The present invention also comprises, the preparation method of Fasudil Hydrochloride crystal formation of the present invention, and the method comprises the following steps: Fasudil Hydrochloride crude product dissolves with anhydrous methanol, adds gac, and insulation is filtered.Filtrate adds anhydrous diethyl ether, and material is separated out, and filters, dry, repeats above step, obtains dry product, twice, water recrystallization.

Preferred preparation method is shown in embodiment.

1 sulfonation

The present invention also comprises,

Fasudil Hydrochloride crystal formation of the present invention and pharmaceutical carrier are made pharmaceutical preparation, and described preparation can be made into any pharmaceutically useful formulation.These formulations comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, paste, sublimed preparation, suspensoid, pulvis, solution, injection, suppository, ointment, plaster, creme, sprays, drops, patch.

Described pharmaceutical carrier comprises any suitable carrier, can be selected from one or more in following vehicle: meglumine, N.F,USP MANNITOL, sorbyl alcohol, Sodium Pyrosulfite, sodium bisulfite, Sulfothiorine, cysteine hydrochloride, Thiovanic acid, methionine(Met), vitamins C, EDETATE SODIUM, Ethylenediaminetetraacetic Acid Calcium Salt, the alkali-metal carbonate of monovalence, acetate, phosphoric acid salt or its aqueous solution, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodium-chlor, Repone K, Sodium.alpha.-hydroxypropionate, Xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, ethanol, Sucralose, citric acid, silicon derivative, Mierocrystalline cellulose and derivative thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, POLYSORBATE 80, agar, calcium carbonate, Calcium hydrogen carbonate, tensio-active agent, polyoxyethylene glycol, cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talcum powder, calcium stearate, Magnesium Stearate, essence, Microcrystalline Cellulose, N.F,USP MANNITOL, hydroxypropylcellulose, sodium starch glycolate.

Wherein in per unit preparation, containing Fasudil Hydrochloride crystal formation of the present invention is 5～500mg.

The preparation method of above-mentioned preparation normally those skilled in that art knows conventional method.

The present invention further discloses this Fasudil Hydrochloride crystal in the application of the relevant various illnesss that cause for the treatment of vasoconstriction.Wherein said relevant various illnesss comprise cerebral embolism, cerebral ischemia, brain injury, vertebrobasilar insufficiency, the caused cerebral vasospasm of subarachnoid hemorrhage.

Experiment situation and the result of fasudil crystal treatment cerebral vasospasm animal model below:

1. experiment material

1.1 medicines and reagent

Fasudil Hydrochloride: white crystals, Tianjin Chasesun Pharmaceutical Co., Ltd provides.With physiological saline, be mixed with desired concn before use.

Nimodipine, yellow powder, Tianjin pharmaceutical factory of central authorities provides.Physiological saline is mixed with desired concn, for experiment in vitro.

Nimotop vial; Tianjin people pharmaceutical factory of amino acid company produces, lot number: 970915, and experiment in coenosarc.

EGTA: Huzhou biological chemistry factory produces, lot number: 881224.

Silica gel microball: chemical reagent two factories in Tianjin provide.

1.2 instrument

Muscular strength transverter: Shanghai Medical Univ produces.

Situation self-balancing recorder: Shanghai Dahua Instrument and Meter Plant.

SXP-1B operating microscope: Shanghai medical optical instrument factory produces.

Rat brain stereotaxic instrument: Xi'an Xi Guang optical modalities agency sells.

Electric pole type tissue blood flow is with RBF-2 type, and Japan manufactures.

1.3 animal

Rat: animal housing provides by Tianjin Inst. of Materia Medica.Laboratory Animal Facility conformity certification " accurate No. 001 of Tianjin Laboratory Animal Facility " is issued by the Tianjin management of laboratory animal council.

Rabbit: New Zealand's kind, is purchased from Tianjin Chun Le animal cultivation factory.

2. test method and result

2.1CaCl ₂cause that isolated rabbit aorta shrinks experiment

Male rabbit, sacrificed by exsanguination, separated aorta, puts sample in Krebs-Henseleit nutritive medium immediately, after endothelium-denuded, sample is cut into the long ring of 0.5cm, and fixedly constant temperature is 37.0 ℃.By muscular strength transverter with desk-top self-balancing recorder record, resting tension 2.0g.Stablize the KCl with 80mM after a hour and cause that aorta shrinks, then by the 80mMK of sample without calcium ⁺nutritive medium wash three times, the EGTA that adds 0.1mM processes 30 minutes, treats that tension force drops to former baseline, accumulative total adds CaCl2, makes that ultimate density is 0.03,0.1,0.3,1,3,10mM makes amount effect curve.In bath, add in advance the Fasudil Hydrochloride of different concns, making ultimate density is 10 ^-7, 3 * 10 ^-7, 10 ^-6with 3 * 10 ^-6m effect repeats the accumulative total effect of CaCl2 after 5 minutes.The nimodipine final concentration of take is made positive control as 1 and 10 μ M, and physiological saline is made blank.

Experimental result shows, CaCl2 shrink tension when larger concentration obviously improves, when Fasudil Hydrochloride concentration is 10 ^-7, 3 * 10 ^-7, 10 ^-6with 3 * 10 ^-6during M, Rabbit Aorta narrowing of the ring tension force all obviously reduces.

The effect of 2.2 pairs of rat brain micro-embolizations

Select 60 of Wistar rats, body weight 394 ± 29g, male, by body weight, be divided at random 6 groups, 10 every group.Be respectively blank group, model group, positive drug nimotop vial 1mg/kg group and Fasudil Hydrochloride 1,3,9mg/kg group.Each organizes the equal abdominal injection 350mg/kg of rat chloral hydrate anesthesia, get dorsal position, general breathing, neck center vertical incision, expose left common carotid, outer and inner carotid artery, the left outside carotid artery of ligation, occipitalia and pterygoid process jaw artery, by left outside carotid artery intubate, enter arteria carotis communis, from 4000 left and right silica gel microballs of Internal carotid artery injection, make it to form Microembolus.Slowly intravenous injection nimodipine (1mg/200g body weight) and Fasudil Hydrochloride 1,3,9mg/kg (0.1mg/100g body weight) immediately after operation, blank group and model group intravenous injection same volume physiological saline.The severity of the rat behavior activity after 24 hours of observation: all comptocormia, circling behavior and rolling behavioral seizures of showing as are typical case's outbreak, press following classification: 3 grades (comptocormia, rotation and rear acroparalysis), 2 grades (after comptocormia, myasthenia of limbs can be creeped), 1 grade (rear myasthenia of limbs can be creeped), 0 (normally).

Experimental result shows, performs the operation after 24 hours, and model group rat behavior is movable obviously abnormal, on average more than 2 grades.The rat behavior progression of intravenous injection Fasudil Hydrochloride 3 and 9mg/kg group obviously reduces, and is dependence with dosage, suitable with the effect of nimodipine 1mg/kg.

The impact of table 2 Fasudil Hydrochloride on Microembolus rat behavior progression

Compare with blank group, ++ P < 0.01

With model group ratio, * P < 0.05, * * P < 0.01

Observe the infarct size of Microembolus after 24 hours: the de-cervical vertebra of rat is put to death, take out brain, be placed in 0 ℃ of physiological saline,-20 ℃ freezing 30 minutes, be cut into 5, the thick 1mm of sheet, 1%TTC37 ℃ of dyeing is after 15 minutes, be placed in 10% formalin solution and fix 24 hours, under operating microscope, calculate the percentage that infarct accounts for the whole ventricles of the brain.

Experimental result shows that 24 hours cerebral Infarction areas of rat micro-embolization account for 46.5% of normal cerebral tissue, after intravenous injection Fasudil Hydrochloride 1,3,9mg/kg, cerebral embolism area obviously reduces, and is dependence with dosage, suitable with nimodipine 1mg/kg effect.

Table 3

Compare with blank group, ++ P < 0.01

With model group ratio, * * P < 0.01

Compare ##P < 0.01 with nimodipine

Observe Microembolus after 24 hours, the degree of cerebral edema: cerebral tissue takes out the rear weight in wet base that claims, after slice dyeing, is placed in far-infrared oven, toasts and weighs after 5 hours, calculates the weight differential of the dry front and back of cerebral tissue, is water-content.

24 hours tissues following MCAO in rats moisture of rat brain micro-embolization obviously increases, and compares with blank, has notable difference.Intravenous injection Fasudil Hydrochloride 9mg/kg tissues following MCAO in rats moisture obviously reduces.

Table 4

Compare+P < 0.05 with blank group

With model group ratio, * * P < 0.01

2.3 impacts on following experimental subarachnoid hemorrhage rat regional cerebral blood flow

Select 58 of Wistar rats, male, by body weight, be divided at random 6 groups, each treated animal all carries out getting ventricumbent position after intraperitoneal anesthesia with 20% urethane, and head is fixed on stereotaxic instrument.In top midline incision, separating muscle and periosteum.5mm before saw bregma, the other 3mm of center line bores with electronic medical courses in general the bone hole that a diameter is about 1mm in place, under operating microscope, endocranium is needled, PE10 plastics tubing sensing ears line mid point is close to front concave bottom and slips into subarachnoid space 1cm (this length is encircled catheter tip just front portion at Willis), connect syringe pumpback, after seeing limpid cerebrospinal fluid, with dental base acrylic resin powder, conduit is fixed on to bone hole, at 3mm after bregma, the other 3mm of center line is in the boring of intubate homonymy, under microscope, needle endocranium, potential electrode is placed in to the micro-of stereotaxic instrument to be entered on fixture, make slowly electrode tip enter parietal cortex 1mm through bone hole, measure LCBF (rCBF).Model group, nimodipine group and three dosage group rats of Fasudil Hydrochloride were slowly injected subarachnoid space in 1 minute with autologous fresh blood 200 μ l, blank group is injected 200 μ l physiological saline through intubate.Then vena femoralis injection immediately.Model group and blank group injecting normal saline, three dosage groups of nimodipine group and Fasudil Hydrochloride are injected respectively nimodipine 1mg/kg and Fasudil Hydrochloride 1,3,9mg/kg.In 5,10, record in the time of 15,20,25,30,40,50,60 minutes.

Experimental result shows, model group rat is injected after autologous blood, and cerebral blood flow (CBF) reduces at once, in observed 1 hour, compares with blank group, all maintains lower level.After intravenous injection Fasudil Hydrochloride 9mg/kg5 minute, cerebral blood flow (CBF) rebound significantly, and maintain about 1 hour, suitable with the effect of nimodipine 1mg/kg; 3mg/kg group injection 5-25 minute, the injection of 1mg/kg group, after 25-30 minute, all have increasing action to rat brain volume of blood flow.

The stability of crystal formation of the present invention is more stable than prior art.

Table 5 study on the stability

Accompanying drawing explanation

What accompanying drawing 1 represented is the X-ray powder diffraction pattern of fasudil.

What accompanying drawing 2 represented is dsc (DSC) differential thermogram of fasudil.

What accompanying drawing 3 represented is infrared (IR) spectrum of fasudil.

What accompanying drawing 4 represented is fasudil ¹³c NMR spectrogram

It is high that Fasudil Hydrochloride crystal formation of the present invention has bioavailability, and drug effect is remarkable, good stability, and yield is high, purity high.Fasudil Hydrochloride crystal formation of the present invention contributes to selection and the design of drug administration approach, and the determining of pharmaceutical preparation technology parameter, thereby improves pharmaceutical production quality.

Embodiment

The present invention is described in further detail in connection with following experimental example or embodiment, but should be appreciated that following embodiment is only for setting forth and explaining the present invention, and does not limit the scope of the invention.

Embodiment 1

The preparation method of Fasudil Hydrochloride crystal formation, the method comprises the following steps:

In retort, drop into isoquinoline 99.9 14000g, open stirring, be down to after room temperature, the vitriol oil 5000ml by having got ready, slowly adds in retort, makes material be solid state, obtains isoquinoline 99.9 vitriol.In retort, drop into 50%.

Oleum 45000ml, opens stirring, is cooled between 15-20 ℃, and upper step isoquinoline 99.9 vitriol 25kg is slowly added, and the Bi Jixu that feeds intake is incubated 3h, after insulation, continues reaction 48h.In retort, add trash ice to carry out ice solution, stir, sulfonation oily matter is slowly added in retort, stir about 4-5h, filters, and obtains 5-sulfonic acid isoquinoline 99.9 crude product.

Refining: in retort, to drop into 10000ml purified water, stir, add crude product 5-sulfonic acid isoquinoline 99.9 to carry out rising temperature for dissolving, after material dissolves completely, be down to room temperature, stir about 3-4h, filter, with air dry oven (80 ℃), be dried, obtain 5-sulfonic acid isoquinoline 99.9.Yield is 60%-75%.

2 chlorinations

In retort, drop into successively sulfur oxychloride 20000ml, N-N dimethyl formamide 200ml, 5-sulfonic acid isoquinoline 99.9 3600g, stir, be warming up to 76-80 ℃ of backflow, insulation reaction 3h, after completion of the reaction, reclaim under reduced pressure sulfur oxychloride, reclaims and finishes, by in methylene dichloride 6000ml sucting reaction tank, stir well rear filtration, with 15000ml filter wash cake, filter, under vacuum condition, (0.06-0.1MPa), with the dry 10-15h of Calcium Chloride Powder Anhydrous, obtains 5-SULPHURYL CHLORIDE isoquinoline 99.9.Yield is 90%-130%.

3 replace

Extract the dichloromethane solution of 5-SULPHURYL CHLORIDE isoquinoline 99.9: in retort, drop into methylene dichloride 27000ml, stir, drop into 5-SULPHURYL CHLORIDE isoquinoline 99.9 a collection of, sodium bicarbonate 2500g, be cooled to 10-15 ℃, the about 20000ml of purified water is added in retort, after stirring is dissolved material completely, stratification, collected organic layer is with standby.

Free alkali preparation: add methylene dichloride 9000ml in retort, 1,4-Diazesuberane 3600g, stirs, and cools the temperature between 0-8 ℃, start slowly to drip the dichloromethane solution of the 5-SULPHURYL CHLORIDE isoquinoline 99.9 preparing, drip and finish, keep temperature to continue reaction 3h, insulation is finished, add about 20000ml purified water, after stirring is dissolved material completely, stratification, collected organic layer.Carry out reclaim under reduced pressure methylene dichloride, obtain oily matter, add 6000ml trichloromethane to make material dissolution, standby.

4 chromatographies

Take 200-300 order silica gel 3500g, dress post, slowly walks post by the free alkali having prepared, then liquid is carried out to wash-out with trichloromethane 30000ml, collects elutriant, standby.

5 distillation salifies

The elutriant preparing is dropped in retort, and reclaim under reduced pressure trichloromethane, adds anhydrous methanol 6000ml, makes material dissolution, slowly drips concentrated hydrochloric acid and adjusts PH to 4.5-6.0.Anhydrous methanol is reclaimed in underpressure distillation, and distillation is finished, and adds anhydrous methanol 6000ml to stir, suck in storage tank, under agitation slowly add anhydrous diethyl ether 24000ml, separate out material, filter, with air dry oven (80 ℃), be dried, obtain Fasudil Hydrochloride crude product.Yield is 55%-70%.

6 is refining

6.1 primary purification

In retort, drop into successively Fasudil Hydrochloride crude product, anhydrous methanol 30000ml, stir and be warming up to 60-65 ℃ of dissolving, material is dissolved completely, add medicinal carbon, be incubated 0.5 hour, filter.

Cold analysis:

Filtrate is filtered in retort, is cooled to below 10 ℃, under agitation, slowly add anhydrous diethyl ether 30000ml, material is separated out, filter, with air dry oven (80 ℃), be dried, obtain primary purification product.

6.2 secondary refining

In retort, drop into primary purification product, chromatogram methyl alcohol 30000ml, stir and be warming up to 60-65 ℃ of dissolving, after material is dissolved completely, add medicinal carbon, be incubated 0.5 hour, filter.

Cold analysis:

Filtrate is filtered in retort, is cooled to below 10 ℃, under agitation, drip anhydrous diethyl ether 30000ml, material is separated out, filter, dry, obtain Fasudil Hydrochloride fine work.Yield is 60%-80%.

Get this product 0.30g, add after purified water 10ml dissolving, solution should be clarified colourless, as occurs muddiness, repeats secondary refining.

7 finished product total recovery scope: 35%-45%.

Claims

1. a preparation method for the crystal formation of Fasudil Hydrochloride, the method step is as follows:

In retort, drop into isoquinoline 99.9 14000g, open stirring, be down to after room temperature, by the vitriol oil 5000ml having got ready, slowly add in retort, make material be solid state, obtain isoquinoline 99.9 vitriol, in retort, drop into 50% oleum 45000ml, open stirring, be cooled between 15-20 ℃, upper step isoquinoline 99.9 vitriol 25kg is slowly added, the Bi Jixu that feeds intake is incubated 3h, after insulation, continue reaction 48h, in retort, add trash ice to carry out ice solution, stir, sulfonation oily matter is slowly added in retort, stir 4-5h, filter, obtain 5-sulfonic acid isoquinoline 99.9 crude product,

Refining: in retort, to drop into 10000ml purified water, stir, add crude product 5-sulfonic acid isoquinoline 99.9 to carry out rising temperature for dissolving, after material dissolves completely, be down to room temperature, stir 3-4h, filter 80 ℃, with air dry oven, be dried, obtain 5-sulfonic acid isoquinoline 99.9, yield is 60%-75%

Chlorination

In retort, drop into successively sulfur oxychloride 20000ml, N-N dimethyl formamide 200ml, 5-sulfonic acid isoquinoline 99.9 3600g, stir, be warming up to 76-80 ℃ of backflows, insulation reaction 3h, after completion of the reaction, reclaim under reduced pressure sulfur oxychloride, reclaim and finish, by methylene dichloride 6000ml sucting reaction tank, stir well rear filtration, with 15000ml filter wash cake, filter, under vacuum condition, with the dry 10-15h of Calcium Chloride Powder Anhydrous, obtain 5-SULPHURYL CHLORIDE isoquinoline 99.9, yield is 90%-130%, and wherein vacuum pressure is-0.06 arrive-0.1MPa;

Replace

Extract the dichloromethane solution of 5-SULPHURYL CHLORIDE isoquinoline 99.9: in retort, drop into methylene dichloride 27000ml, stir, drop into 5-SULPHURYL CHLORIDE isoquinoline 99.9 a collection of, sodium bicarbonate 2500g, be cooled to 10-15 ℃, purified water 20000ml is added in retort, after stirring is dissolved material completely, stratification, collected organic layer is with standby;

Free alkali preparation: in retort, add methylene dichloride 9000ml, 1,4-Diazesuberane 3600g, stir, cool the temperature between 0-8 ℃, start slowly to drip the dichloromethane solution of the 5-SULPHURYL CHLORIDE isoquinoline 99.9 preparing, drip and finish, keep temperature to continue reaction 3h, insulation is finished, and adds 20000ml purified water, after stirring is dissolved material completely, stratification, collected organic layer, carry out reclaim under reduced pressure methylene dichloride, obtain oily matter, add 6000ml trichloromethane to make material dissolution, standby;

Chromatography

Take 200-300 order silica gel 3500g, dress post, slowly walks post by the free alkali having prepared, then liquid is carried out to wash-out with trichloromethane 30000ml, collects elutriant, standby;

Distillation salify

The elutriant preparing is dropped in retort, and reclaim under reduced pressure trichloromethane, adds anhydrous methanol 6000ml, make material dissolution, slowly drip concentrated hydrochloric acid and adjust PH to 4.5-6.0, anhydrous methanol is reclaimed in underpressure distillation, distillation is finished, and adds anhydrous methanol 6000ml to stir, and sucks in storage tank, under agitation slowly add anhydrous diethyl ether 24000ml, separate out material, filter, with air dry oven, be dried, temperature is 80 ℃, obtains Fasudil Hydrochloride crude product, and yield is 55%-70%;

Refining

Primary purification

In retort, drop into successively Fasudil Hydrochloride crude product, anhydrous methanol 30000ml, stir and be warming up to 60-65 ℃ of dissolvings, material is dissolved completely, add medicinal carbon, be incubated 0.5 hour, filter;

Cold analysis:

Filtrate is filtered in retort, is cooled to below 10 ℃, under agitation, slowly add anhydrous diethyl ether 30000ml, material is separated out, filter, with air dry oven, be dried, temperature is 80 ℃, obtains primary purification product;

Secondary refining

In retort, drop into primary purification product, chromatogram methyl alcohol 30000ml, stir and be warming up to 60-65 ℃ of dissolvings, after material is dissolved completely, add medicinal carbon, be incubated 0.5 hour, filter;

Cold analysis:

Filtrate is filtered in retort, is cooled to below 10 ℃, under agitation, drip anhydrous diethyl ether 30000ml, material is separated out, filter, dry, obtain Fasudil Hydrochloride fine work, yield is 60%-80%;

Get this product 0.30g, add after purified water 10ml dissolving, solution should be clarified colourless, as occurs muddiness, repeats secondary refining, obtains the crystal formation of Fasudil Hydrochloride, finished product total recovery scope: 35%-45%;

The crystal formation of Fasudil Hydrochloride has following spectrum characteristic, adopt Rigaku Rigaku D max-2500 type X powder diffraction instrument to analyze the crystalline phase of sample, Cu K α target, tube voltage 40KV, tube current 100mA, its X-powdery diffractometry has following characteristic peak: at approximately 6.780,14.700,16.620,17.480,20.440,20.740,24.52 degree 2 θ places, have peak.