CN1628790A - Injecta of anemarrhena extractive - Google Patents
Injecta of anemarrhena extractive Download PDFInfo
- Publication number
- CN1628790A CN1628790A CN 200410054146 CN200410054146A CN1628790A CN 1628790 A CN1628790 A CN 1628790A CN 200410054146 CN200410054146 CN 200410054146 CN 200410054146 A CN200410054146 A CN 200410054146A CN 1628790 A CN1628790 A CN 1628790A
- Authority
- CN
- China
- Prior art keywords
- rhizoma anemarrhenae
- anemarrhenae extract
- injection
- extract
- water
- 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
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an injection of anemarrhena extractive, which includes the various dosage types of powder injection, small injection, glucose fluid infusion and sodium chloride fluid infusion, wherein the main active constituent of anemarrhena extract includes anemarrhena saponin B-II, anemarrhena saponin E1, anemarrhena saponin B by the weight ratio of 78-92 : 2-12 : 0-6.
Description
Technical field:
The present invention relates to medical technical field, is the injection type of Rhizoma Anemarrhenae extract, comprises injectable powder, small-volume injection, glucose infusion liquid or sodium chloride transfusion, also relates to the new purposes of its treatment ischemic cerebrovascular.
Background technology:
The Rhizoma Anemarrhenae is a liliaceous plant Rhizoma Anemarrhenae Anemarrhena asphodeloides Bunge rhizome, the effect that has nourishing YIN to lower pathogenic fire, moisturizes laxation.
The chemical constituent of the Rhizoma Anemarrhenae is mainly saponin, flavones ingredient.The timosaponin constituents has the alzheimer disease effect that prevents and treats significantly (application for a patent for invention prospectus, publication number CN 1301166, application number 99806158.1; Application for a patent for invention prospectus, publication number CN 1212966, application number 97119680.X), the myocardium I/R damage of protection (Capital Medical College's journal, 1994; 138), platelet aggregation inhibitory activity (Clin Chim Acta, 1,999 15 (2):; 289 (1-2): 79-88), the effect (application for a patent for invention prospectus, publication number CN 1370537, application number 01106195.2) that produces of estrogen-like function of resisting osteoporosis, antiinflammatory, inhibition neutrophilic granulocyte peroxide etc.; With timosaponin B-II, timosaponin E
1, timosaponin E, timosaponin B, 1-timosaponin A-1 III etc. have treatment apoplexy activity for the Rhizoma Anemarrhenae extract of main component, can obviously improve behavior symptom that cerebral ischemia causes, dwindle cerebral infarct volume, reduce the rats with cerebral ischemia cerebral edema, medicine (the application for a patent for invention prospectus that can be used for preparation control apoplexy (apoplexy), publication number CN 1451384A, application number 03116824.8).
Be mainly the single Rhizoma Anemarrhenae decoction pieces when at present the Rhizoma Anemarrhenae is medicinal, oral after frying in shallow oil soup or be prepared into solid preparation with other drug matchings.Because oral have defectives such as curative effect is slow, the effective ingredient bioavailability is low, can not satisfy the treatment needs of cardiovascular and cerebrovascular disease faster of falling ill, especially also there is dysphagia in the patients with cerebral apoplexy morbidity at acute stage, therefore, if with the active substance of the Rhizoma Anemarrhenae-with the saponin is that the Rhizoma Anemarrhenae extract of main component is prepared into the deficiency that injection type just can overcome oral formulations, be specially adapted to the treatment of cardiovascular and cerebrovascular disease.But do not see the report of relevant Rhizoma Anemarrhenae extract injection type at present, do not see that the Rhizoma Anemarrhenae is used as the report of treatment ischemic cerebrovascular medicine yet.
Summary of the invention:
The present invention is prepared into injection type with Rhizoma Anemarrhenae extract, comprises injectable powder, small-volume injection, glucose infusion liquid or sodium chloride transfusion, and the new purposes that said preparation is used for the treatment of ischemic cerebrovascular also is provided simultaneously.
Rhizoma Anemarrhenae extract injection of the present invention is made up of Rhizoma Anemarrhenae extract, pharmaceutic adjuvant, and pharmaceutic adjuvant is selected from excipient, osmotic pressure regulator, diluent etc.
The preparation method of the said Rhizoma Anemarrhenae extract of the present invention is: routinely the rhizoma ane marrhenae of pulverizing is carried with 20～95% pure heat, the solvent volumetric usage is about 4～8 times of crude drug first; Maybe with the rhizoma ane marrhenae pulverized with 20～95% ethanol percolate extraction, the solvent volumetric usage is about 4～12 times of crude drug; Extracting solution concentrates, the centrifugal or filtration of concentrated solution; (nonpolar or low pole macroporous resin is for being the polystyrene type porous adsorbent resin of bridging materials with styrene by nonpolar or low pole macroporous resin adsorption for the centrifuged supernatant of concentrated solution or filtrate, as D101, D201, ZTC-1, AB-8,1300-1 type macroporous resin etc.), impurity is removed in water and 20% following ethanol drip washing; The moisture lower alcohol eluting of reuse, lower alcohol is C such as methanol, ethanol or propanol
1～C
5Alcohols, its concentration are 30～95%, and volumetric usage is 5～15 times of crude drug, collect the lower alcohol eluent, and being concentrated into does not have alcohol; Concentrated solution again by the macroporous resin adsorption eluting, is collected the lower alcohol eluent as stated above, and being concentrated into does not have alcohol, and crushed after being dried or spray drying, lyophilization are Rhizoma Anemarrhenae extract of the present invention; Or with concentrated solution by the polyamide chromatograph, with water elution, collect water elution liquid, concentrate, crushed after being dried or spray drying, lyophilization also can get Rhizoma Anemarrhenae extract of the present invention.
Contain effective composition timosaponin B-II, timosaponin E in the Rhizoma Anemarrhenae extract of the present invention
1, timosaponin B sum accounts for total amount more than 80%, timosaponin B-II, timosaponin E
1, timosaponin B the percentage composition example be 78～92: 2～12: 0～6.
The present invention prepares injectable powder and generally adopts conventional freeze-drying, as solvent, the steps include: to get Rhizoma Anemarrhenae extract with water, adds excipient, is dissolved in water, and adds active carbon, filtration sterilization, and plug is partly rolled in fill, and lyophilization, tamponade are rolled lid and are got final product.Used excipient is selected from one or more in mannitol, gelatin hydrolysate, glucose, lactose, the dextran etc.Every bottle contains Rhizoma Anemarrhenae extract 5～50mg.
The present invention prepares injectable powder also can adopt spray drying method, as solvent, the steps include: to get Rhizoma Anemarrhenae extract with water, adds or do not add excipient (excipient is the same), be dissolved in water, add active carbon, filtration sterilization, spray drying, aseptic subpackaged, tamponade is rolled lid and is got final product.Every bottle contains Rhizoma Anemarrhenae extract 5～50mg.
When the present invention prepared small-volume injection, preparation got final product as solvent with water for injection, also can add appropriate amount of auxiliary materials, and adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.Every contains Rhizoma Anemarrhenae extract 5～50mg.
The present invention prepares glucose infusion liquid or sodium chloride transfusion, with water for injection as solvent, add the preparation of an amount of glucose or sodium chloride and get final product, also can add appropriate amount of auxiliary materials, adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.Every bottle contains Rhizoma Anemarrhenae extract 5～50mg.
The present invention has carried out isolation identification to the chemical constituent of Rhizoma Anemarrhenae extract.
Separation and purification: get Rhizoma Anemarrhenae extract 10g noted earlier,, cross sephadex LH20 column chromatography, in varing proportions H with dissolved in distilled water
2O-Me
2The CO eluting.Collect H
2O-Me
2CO (2: 1) eluting part, drying gets dry thing 0.6g, is component 1.Collect H
2O-Me
2CO (1: 1) eluting part, drying gets dry thing 7.8g, is component 2.Collect H
2O-Me
2CO (1: 2) eluting part, drying gets dry thing 0.7g, is component 3.
Get 3 each 500mg of component, use H respectively
2The O dissolving is carried out the medium pressure liquid chromatography separation and purification, chromatographic condition: the anti-phase preparative column of LobarC18, H in varing proportions
2O-Me
2CO makes mobile phase, and reverse phase silica gel TLC detects, and collects out peak stream part.The mobile phase of component 1 is H
2O-Me
2CO (70: 30, V: V), get white powder 125mg, be chemical compound 1.The mobile phase of component 2 is H
2O-Me
2CO (60: 40, V: V), get white powder 396mg, be chemical compound 2.The mobile phase of component 3 is H
2O-Me
2CO (35: 65, V: V), get white powder 183mg, be chemical compound 3.
Structure is identified: integrated application UV, IR, and MS,
1H-NMR,
13C-NMR, the 2D-NMR analytical technology is identified structure.Chemical compound 1,2,3 is accredited as timosaponin E respectively
1, timosaponin B-II, timosaponin B.Their chemical constitution is as follows:
Timosaponin E
1The structural formula of structural formula timosaponin B-II
The structural formula of timosaponin B
Timosaponin E
1The evaluation white powder, mp164～167 ℃; IR (KBr, cm
-1): 3418 (OH), 2929 (CH
2), 1455 (CH
2), 1076 (C-O-C); ESI-MSm/z:936;
1H-NMR (C
5D
5N, δ, ppm): 5.33 (1H, d, J=8Hz, Glc-1H), 4.89 (1H, d, J=8Hz, Gal-1H), 4.82 (1H, d, J=8Hz, Glc-1H), 1.32 (3H, d, 21-CH
3), 1.03 (3H, d, 27-CH
3), 1.03 (3H, s, 19-CH
3), 0.91 (3H, s, 18-CH
3);
13CNMR (C
5D
5N, δ): see Table 1.
The evaluation white powder of timosaponin B-II, mp246～248 ℃; IR (KBr, cm
-1): 3424 (OH), 2929 (CH
2), 1450 (CH
2), 1076 (C-O-C); ESI-MSm/z:920;
1H-NMR (C
5D
5N, δ, ppm): 5.28 (1H, d, J=7Hz, Glc-1H), 4.91 (1H, d, J=8Hz, Gal-1H), 4.80 (1H, d, J=8Hz, Glc-1H), 1.32 (3H, d, 21-CH
3), 1.03 (3H, d, 27 CH
3-), 0.98 (3H, s, 19-CH
3), 0.88 (3H, s, 18-CH
3);
13CNMR (C
5D
5N, δ): see Table 2.Structural formula is seen Fig. 2.
The evaluation white powder of timosaponin B, mp＞226 ℃ (dec); IR (KBr) cm
-1: 3385 (OH), 2921 (CH
2), 1712 (C=C), 1451 (CH
2), 1074 (C-O-C); ESI-MSm/z:902;
1HNMR (DMSO-d
6, δ): δ 0.61 (18-CH
3, 3H, s), 0.87 (27-CH
3, 3H, s), 1.55 (21-CH
3, 3H, s), 0.89 (19-CH
3, 3H, d), δ 4.08 (1 -H, 1H, d, J=8Hz), 4.27 (1 '-H, 1H, d, J=7Hz), 4.41 (1 " H, 1H, d, J=8Hz);
13CNMR (DMSO-d
6, δ): see Table 3.
Table 1 timosaponin E
1 13C-NMR data (δ, ppm, C
5D
5N)
C δ chemical type C δ chemical type
1 30.8 CH
2 26 75.0 CH
2
2 26.8 CH
2 27 17.7 CH
3
3 75.0 CH galactose
4 30.6 CH
2 1 102.1 CH
5 36.7 CH 2 81.4 CH
6 26.7 CH
2 3 76.7 CH
7 26.4 CH
2 4 69.5 CH
8 35.2 CH 5 76.5 CH
9 40.3 CH 6 62.5 CH
2
10 35.0 C Glucose
11 20.8 CH
2 1 105.7 CH
12 40.5 CH
2 2 75.1 CH
13 40.9 C 3 78.1 CH
14 60.5 CH 4 71.4 CH
15 78.7 CH 5 78.3 CH
16 91.1 CH 6 62.5 CH
2
17 61.1 CH Glucose
18 16.4 CH
3 1′ 104.8 CH
19 23.9 CH
3 2′ 74.9 CH
20 40.1 CH 3′ 78.0 CH
21 16.2 CH
3 4′ 71.4 CH
22 110.0 C 5′ 77.7 CH
23 36.8 CH
2 6′ 61.0 CH
2
24 28.0 CH
2
25 34.1 CH
Table 2 timosaponin B-II's
13C-NMR data (δ, ppm, C
5D
5N)
C δ chemical type C δ chemical type
1 30.6 CH
2 26 75.1 CH
2
2 26.7 CH
2 27 17.2 CH
3
3 74.8 CH galactose
4 30.6 CH
2 1 102.2 CH
5 36.6 CH 2 81.4 CH
6 26.5 CH
2 3 76.2 CH
7 26.5 CH
2 4 69.5 CH
8 35.2 CH 5 76.5 CH
9 40.3 CH 6 62.5 CH
2
10 34.9 C Glucose
11 20.9 CH
2 1 105.7 CH
12 40.1 CH
2 2 75.2 CH
13 40.9 C 3 78.0 CH
14 56.1 CH 4 71.3 CH
15 32.1 CH
2 5 78.3 CH
16 80.9 CH 6 62.5 CH
2
17 63.7 CH Glucose
18 16.4 CH
3 1′ 104.8 CH
19 23.7 CH
3 2′ 74.9 CH
20 40.1 CH 3′ 78.1 CH
21 16.1 CH
3 4′ 71.4 CH
22 110.4 C 5′ 77.7 CH
23 36.8 CH
2 6′ 61.9 CH
2
24 28.0 CH
2
25 34.1 CH
Table 3 timosaponin B's
13C-NMR data (δ, ppm, DMSO-d
6)
C δ chemical type C δ chemical type
1 30.1 CH
2 26 73.8 CH
2
2 26.4 CH
2 27 16.7 CH
3
3 73.7 CH galactose
4 29.8 CH
2 1 103.2 CH
5 35.9 CH 2 78.9 CH
6 26.4 CH
2 3 73.3 CH
7 25.9 CH
2 4 67.9 CH
8 34.6 CH 5 73.0 CH
9 39.7 CH 6 61.0 CH
2
10 34.0 C Glucose
11 34.5 CH
2 1 103.6 CH
12 39.4 CH
2 2 74.6 CH
13 43.2 C 3 76.6 CH
14 54.1 CH 4 70.0 CH
15 30.5 CH
2 5 76.8 CH
16 83.5 CH 6 61.0 CH
2
17 63.7 CH Glucose
18 14.0 CH
3 1′ 100.6 CH
19 23.5 CH
3 2′ 74.9 CH
20 102.8 C 3′ 75.9 CH
Through animal experiment, the quiet notes administration of Rhizoma Anemarrhenae extract of the present invention can obviously improve behavior symptom that cerebral ischemia causes, dwindles cerebral infarct volume, reduce the rats with cerebral ischemia cerebral edema.In addition, the quiet notes administration of Rhizoma Anemarrhenae extract can also the significant prolongation mice broken end back mouth breathing persistent period, increase anesthetized dog cerebral blood flow, suppress rabbit thrombosis, reduce rabbit whole blood viscosity etc.Show that with Rhizoma Anemarrhenae extract of the present invention be the injection type that feedstock production forms, can also be applied to the treatment of ischemic cerebrovascular.
The invention provides a kind of pharmaceutical preparation of Rhizoma Anemarrhenae extract, is that Rhizoma Anemarrhenae extract is prepared into injection type.The pharmacological action of Rhizoma Anemarrhenae extract injection is stronger, and its raw material sources are abundant, inexpensive, and preparation technology is simple.
Description of drawings
Fig. 1 is the chromatogram of Rhizoma Anemarrhenae extract of the present invention.
The specific embodiment
Below in conjunction with embodiment the present invention is described in detail.
The preparation of embodiment 1 Rhizoma Anemarrhenae extract
Rhizoma ane marrhenae is thinly sliced, got 10kg,, collect the 240L percolate with 30% ethanol 30L infiltration percolation extraction after 12 hours.Percolate is concentrated to 15L; Concentrated solution filters, and filtrate feeding is equipped with in the chromatographic column of D101 type resin of 20L, uses distilled water 300L, 20% ethanol 300L, 60% ethanol 120L eluting successively, collects 60% ethanol elution part; 60% ethanol elution concentrating under reduced pressure feeds in the chromatographic column of the D101 type resin that 20L is housed to 10L; Use 20% ethanol 300L, 60% ethanol 120L eluting successively, collect 60% ethanol elution part, concentrating under reduced pressure, drying gets faint yellow crystalloid powder, is Rhizoma Anemarrhenae extract of the present invention, and heavy altogether 0.14kg is 1.4% by crude drug weight yield.
The preparation of embodiment 2 Rhizoma Anemarrhenae extracts
Rhizoma ane marrhenae is thinly sliced, get 10kg, carry 3 times with 50% ethanol 50L heat successively at 90～100 ℃, each extraction time is 90 minutes, merges 3 times extracting solution, is concentrated into 20L, filter, filtrate feeds in the chromatographic column of the ZTC-1 type resin that 25L is housed, and uses distilled water 500L, 20% ethanol 250L, 50% ethanol 150L eluting successively, collects 50% ethanol elution part; 50% ethanol elution concentrating under reduced pressure feeds and is equipped with in the chromatographic column of 10kg polyamide to 8L, uses the distilled water eluting, collect distilled water eluent 30L, concentrating under reduced pressure, spray drying gets pale yellow powder, be Rhizoma Anemarrhenae extract of the present invention, heavy altogether 0.17kg is 1.7% by crude drug weight yield.
The preparation of embodiment 3 injectable powder
Get Rhizoma Anemarrhenae extract 30g, add dextran 30g, add 500ml water for injection, stir and make its dissolving; Add the injection water to 2000ml, add the 3.0g needle-use activated carbon, fully stirred 30 minutes; Decarbonization filtering; With 0.22 μ m filtering with microporous membrane; Be filled in the aseptic cillin bottle, every bottle of 2ml partly rolls plug; Lyophilization, tamponade is rolled lid and is got final product again.
The preparation of embodiment 4 injectable powder
Get Rhizoma Anemarrhenae extract 15g, add mannitol 40g, add 500ml water for injection, stir and make its dissolving; Add the injection water to 1000ml, add the 1g needle-use activated carbon, fully stirred 30 minutes; Decarbonization filtering; With 0.22 μ m filtering with microporous membrane; Lyophilization gets sterilized powder, is distributed into 1000 bottles.
The preparation of embodiment 5 injectable powder
Get Rhizoma Anemarrhenae extract 20g, add lactose 50g, add 100ml water for injection, stir and make its dissolving; Add the injection water to 1000ml, add the 1.5g needle-use activated carbon, fully stirred 30 minutes; Decarbonization filtering; With 0.22 μ m filtering with microporous membrane; Spray drying gets sterilized powder, is distributed into 1000 bottles.
The preparation of embodiment 6 small-volume injections
Get Rhizoma Anemarrhenae extract 1g, add 100ml water for injection, stir and make its dissolving; Add the injection water to 1000ml, with 0.22 μ m filtering with microporous membrane; The packing embedding, every bottle of 10ml, sterilization gets final product.
The preparation of embodiment 7 small-volume injections
Get Rhizoma Anemarrhenae extract 5g, add propylene glycol 20g, add 200ml water for injection, stir and make its dissolving; Add the injection water to 1000ml, add the 1.5g needle-use activated carbon, fully stirred 30 minutes; Decarbonization filtering; With 0.22 μ m filtering with microporous membrane; The packing embedding, every bottle of 5ml, sterilization gets final product.
The preparation of embodiment 8 glucose infusion liquids
Get Rhizoma Anemarrhenae extract 1g, add Polyethylene Glycol 10g, add glucose 500g, add 2000ml water for injection, stir and make its dissolving; Add the injection water to 5000ml; With 0.22 μ m filtering with microporous membrane; The packing embedding, every bottle of 100ml, sterilization gets final product.
The preparation of embodiment 9 glucose infusion liquids
Get Rhizoma Anemarrhenae extract 1g, add glucose 250g, add 1000ml water for injection, stir and make its dissolving; Add the injection water to 5000ml; With 0.22 μ m filtering with microporous membrane; The packing embedding, every bottle of 250ml, sterilization gets final product.
The preparation of embodiment 10 sodium chloride transfusion
Get Rhizoma Anemarrhenae extract 1g, add sodium chloride 90g, add 1000ml water for injection, stir and make its dissolving; Add the injection water to 10000ml; With 0.22 μ m filtering with microporous membrane; The packing embedding, every bottle of 250ml, sterilization gets final product.
The assay of the main chemical compositions of Rhizoma Anemarrhenae extract of the present invention adopts high performance liquid chromatography to carry out.
Chromatographic condition Waters high performance liquid chromatograph, 510 pumps; The SEDEX of Di Ma company 75 type evaporative light scattering detector (ELSD); SR2000 work station (going up sharp Science and Technology Ltd. of naval); Dalian Yi Lite hypersil ODS post (4.6 * 250mm, 5 μ) chromatographic column; Mobile phase is acetonitrile-water (30: 70); Flow velocity is 0.8mL/min, nebulizer gas pressure 3.5bar, 40 ℃ of drift tube temperatures, GAIN=7.
Algoscopy
1. the preparation acetonitrile-aqueous solution is with acetonitrile: water=30: 70 is mixed with acetonitrile-aqueous solution
2. be solvent with the acetonitrile-aqueous solution, prepare following solution:
(1) timosaponin B-II reference substance solution, concentration 250 μ g/mL;
(2) timosaponin E
1Reference substance solution, concentration 25 μ g/mL;
(3) timosaponin B reference substance solution, concentration 10 μ g/mL;
(4) Rhizoma Anemarrhenae extract solution of the present invention, concentration 300 μ g/mL.
3. detect and get reference substance solution and each 20 μ L of need testing solution respectively, inject chromatograph of liquid, the record chromatogram, chromatogram is seen Fig. 1.Calculate by the logarithm of external standard method, promptly with peak area.
Through repeated detection, timosaponin B-II, timosaponin E in the Rhizoma Anemarrhenae extract
1, timosaponin B the percentage composition example be 78～92: 2～12: 0～6; Timosaponin B-II, timosaponin E in the Rhizoma Anemarrhenae extract
1, timosaponin B the content sum more than 80%.
The present invention tests the protective effect of SD (Sprague-Dawley) cerebral ischemia by intravenous injection Rhizoma Anemarrhenae extract.
Test method:
SD rat (Shanghai Branch of Chinese Academy of Sciences animal center provides) is divided into 5 groups at random: sham operated rats, negative control group, positive controls, Rhizoma Anemarrhenae extract low dose group and Rhizoma Anemarrhenae extract high dose group, every group 30, male and female half and half, body weight 180～230g.Sham operated rats is the one-sided common carotid artery of ligation only, to get rid of the influence of operation to experimental result itself; Negative control group is the thromboembolism intraluminal middle cerebral artery occlusion in rats, pours into 24 hours damage models in 2 hours again as focal cerebral ischemia, gives the normal saline of equivalent; Positive control Ginaton injection (powder pin, Weimar, Germany doctor Shu Pei pharmaceutical factory, lot number 5530701), it recommends clinical mean dose is 2.0mg/kg/ people/day (people presses the 60kg body weight and calculates), conversion is 12mg/kg for the dosage of rat, design positive drug dosage quite is subjected to the middle dosage of reagent thing, so be taken as 36mg/kg.The Rhizoma Anemarrhenae extract low dose group is 1.8mg/kg, and basic, normal, high dosage ratio is 1: 3: 6, and corresponding middle and high dosage is respectively 5.4mg/kg and 11mg/kg.The all quiet notes administration of each group.
(preparation method sees pharmacology's part of " Western medicine new drug preclinical study guideline compilation ", bureau of drug policy ﹠ administration of Ministry of Health of the People's Republic of China, 1993:73 for details to prepare SD cerebral ischemic reperfusion in rats model with the bolt collimation method; " pharmacological experimental methodology " second edition, People's Health Publisher, 1982:830,1113), except that sham operated rats, all the other each groups are all with this model mouse test.Each group is in ischemia quiet notes administration after 1 hour, and ischemia poured into after 1 hour 24 hours more again, observed following index then:
(1) neurological is influenced: observe the survival Mus after 24 hours behavior change, carry out behavioristics's scoring, with reference to 5 fens system standards of grading of Zea Longa: 0 be divided into normal, impassivity damage symptom; 1 be divided into can not full extension offside fore paw; 2 are divided into laterally and turn-take; 3 are divided into to offside and topple over; 4 are divided into and can not spontaneously walk loss of consciousness.
(2) to the influence of brain water content: every group get 8 rats fast broken end get brain, divide another name left and right sides brain hemisphere weight in wet base, put oven dry in 160 ℃ of baking boxs, claim dry weight after 24 hours, calculate brain water content as follows: brain water content (%)=(weight in wet base-dry weight)/weight in wet base * 100%.
(3) to the influence of cerebral infarct volume: get 8 rats for every group, put to death and get brain, downcut the crown brain sheet of thick about 2mm, place 2% triphenyltetrazolium chloride solution at once, hatched 30 minutes for 37 ℃.Infarct presents white, and non-infarct presents redness.Digital camera is taken record, surveys brain sheet cumulative volume and infarct volume with Medbrain 2.0 softwares (Nanjing Mei Yi scientific ﹠ technical corporation), calculates the percentage ratio (%) that infarct accounts for whole cerebral tissue.
Result of the test:
(1) each group of neurological influence is got 10 rats at random, observe the behavior of survival Mus.The result sees table 4 for details.
Table 4 Rhizoma Anemarrhenae extract lacks neurological that 2h the pours into 24h again influence (x ± SD) that marks to rat brain
Group dosage number of animals (n) behavior scoring
Sham operated rats 4ml/kg 10 0
Solvent control group 4ml/kg 10 1.7 ± 0.67
Rhizoma Anemarrhenae extract 1.8mg/kg 10 1.2 ± 0.92
Rhizoma Anemarrhenae extract 5.4mg/kg 10 0.9 ± 0.32
*
Rhizoma Anemarrhenae extract 11.0mg/kg 10 0.6 ± 0.52
*
Ginaton injection 36mg/kg 10 0.6 ± 0.70
*
Compare with the solvent control group
*P＜0.05,
*P＜0.01
As seen from Table 4, the middle and high dosage group of Rhizoma Anemarrhenae extract all can significantly reduce behavior scoring, with the solvent control group significant difference (P＜0.05, P＜0.01) is arranged relatively, and the reduction degree has the doses dependence.Illustrate that the quiet notes of Rhizoma Anemarrhenae extract can obviously improve the nerve injury symptom that causes because of ischemia-reperfusion.
(2) experimental result that influences to brain water content sees Table 5.
Table 5 Rhizoma Anemarrhenae extract is to the influence of Ischemia and Reperfusion in vivo in Rats brain water content (x ± D)
Group dosage number of animals (n) left hemisphere water content (%) right hemisphere water content (%)
Sham operated rats 4ml/kg 8 80.52 ± 0.91 80.52 ± 0.95
*
Solvent control group 4ml/kg 8 80.48+0.48 85.46 ± 1.18
△ △
Rhizoma Anemarrhenae extract 1.8mg/kg 8 81.02 ± 0.74 83.34 ± 1.81
△ △ * *
Rhizoma Anemarrhenae extract 5.4mg/kg 8 80.81 ± 0.67 82.31 ± 2.51
*
Rhizoma Anemarrhenae extract 11.0mg/kg 8 80.81 ± 0.67 81.46 ± 0.60
*
Ginaton injection 36mg/kg 8 80.80 ± 0.87 81.87 ± 1.35
*
Compare with the solvent control group,
*Compare with sham operated rats P＜0.01,
△ △P＜0.01
As seen from Table 5, the brain water content of solvent control normal saline group ischemia side (right hemisphere) is significantly higher than sham operated rats (P＜0.01), the brain water content of Rhizoma Anemarrhenae extract low dose group is significantly higher than sham operated rats (P＜0.01), but be lower than the solvent control group, significant difference (P＜0.01) is arranged between the two; The brain water content of middle and high dosage group of Rhizoma Anemarrhenae extract and Ginaton injection group significantly is lower than solvent control group (P＜0.01), with the sham operated rats there was no significant difference.The result shows that the quiet notes of Rhizoma Anemarrhenae extract can significantly reduce the water content of Ischemia and Reperfusion in vivo in Rats injured brain tissue, alleviates ischemia side brain hemisphere edema degree.
(3) experimental result that influences to cerebral infarct volume sees Table 6.
Table 6 Rhizoma Anemarrhenae extract is to the influence of the cerebral infarct volume of cerebral ischemic reperfusion in rats (x ± SD)
Group dosage number of animals (n) infarct volume accounts for whole cerebral tissue percentage ratio (%)
Sham operated rats 4ml/kg 80
Solvent control group 4ml/kg 8 31.80 ± 3.63
Rhizoma Anemarrhenae extract 1.8mg/kg 8 21.05 ± 3.83
*
Rhizoma Anemarrhenae extract 5.4mg/kg 8 13.23 ± 4.38
*
Rhizoma Anemarrhenae extract 11mg/kg 8 5.65 ± 5.79
*
Ginaton injection 36mg/kg 8 9.61 ± 7.31
*
Compare with the solvent control group,
*P＜0.01
As seen from Table 6, the sham operated rats cerebral tissue does not have infraction, solvent control group ischemia side cerebral tissue has the infraction phenomenon, the percentage ratio that infarct accounts for whole cerebral tissue is 31.80 ± 3.63%, basic, normal, high 3 dosage groups of Rhizoma Anemarrhenae extract and positive control Ginaton injection group all can significantly be dwindled ischemia side cerebral tissue infarct volume, have compared highly significant difference (P＜0.01) with the solvent control group, have reduced by 33.81% respectively, 58.4%, 82.23% and 69.78%; And high dose group is dwindled the effect of ischemia side cerebral tissue infarct volume and is better than positive control Ginaton injection group.The result shows that the cerebral infarction that the quiet notes of Rhizoma Anemarrhenae extract cause cerebral ischemia is plugged with remarkable protective effect.
Claims (7)
1. the Rhizoma Anemarrhenae extract injection is made up of Rhizoma Anemarrhenae extract, pharmaceutic adjuvant, comprises injectable powder, small-volume injection, glucose infusion liquid and sodium chloride transfusion, and said Rhizoma Anemarrhenae extract is made by following method:
The rhizoma ane marrhenae of pulverizing is carried with 20～95% pure heat, and the solvent volumetric usage is about 4～8 times of crude drug first; Maybe with the rhizoma ane marrhenae pulverized with 20～95% ethanol percolate extraction, the solvent volumetric usage is about 4～12 times of crude drug; Extracting solution concentrates, the centrifugal or filtration of concentrated solution; The centrifuged supernatant of concentrated solution or filtrate are by nonpolar or low pole macroporous resin adsorption, and impurity is removed in water and 20% following ethanol drip washing; The moisture lower alcohol eluting of reuse, lower alcohol is selected from C
1～C
5Alcohols, its concentration are 30～95%, and volumetric usage is 5～15 times of crude drug, collect the lower alcohol eluent, and being concentrated into does not have alcohol; Concentrated solution again by the macroporous resin adsorption eluting, is collected the lower alcohol eluent as stated above, and being concentrated into does not have alcohol, and crushed after being dried or spray drying, lyophilization are Rhizoma Anemarrhenae extract; Or with concentrated solution by the polyamide chromatograph, with water elution, collect water elution liquid, concentrate, crushed after being dried or spray drying, lyophilization also can get Rhizoma Anemarrhenae extract.
2. Rhizoma Anemarrhenae extract injection according to claim 1 is characterized in that effective ingredient timosaponin B-II, the timosaponin E of said Rhizoma Anemarrhenae extract
1, timosaponin B the percentage composition example be 78～92: 2～12: 0～6, the content sum accounts for more than 80% of Rhizoma Anemarrhenae extract gross weight.
3. Rhizoma Anemarrhenae extract injection according to claim 1 and 2, it is characterized in that said injectable powder is to adopt in mannitol, gelatin hydrolysate, glucose, lactose, the dextran one or more to make excipient by Rhizoma Anemarrhenae extract, after lyophilization, packing gets final product as required.
4. Rhizoma Anemarrhenae extract injection according to claim 1 and 2 is characterized in that said injectable powder is to adopt water for injection or ethanol by Rhizoma Anemarrhenae extract, or water for injection and alcoholic acid mixed liquor be as solvent, spray-dried after, packing gets final product as required.
5. Rhizoma Anemarrhenae extract injection according to claim 1 and 2, it is characterized in that said small-volume injection be after adopting water for injection as the solvent preparation by Rhizoma Anemarrhenae extract as required packing get final product; Also can add appropriate amount of auxiliary materials, adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.
6. Rhizoma Anemarrhenae extract injection according to claim 1 and 2 is characterized in that the transfusion of said glucose infusion liquid or sodium chloride is to adopt water for injection as solvent by Rhizoma Anemarrhenae extract, and adding after the preparation of an amount of glucose or sodium chloride as required, packing gets final product; Also can add appropriate amount of auxiliary materials, adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.
7. the application of the described Rhizoma Anemarrhenae extract of claim 1 in preparation treatment ischemic cerebrovascular medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410054146 CN1628790A (en) | 2004-08-31 | 2004-08-31 | Injecta of anemarrhena extractive |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410054146 CN1628790A (en) | 2004-08-31 | 2004-08-31 | Injecta of anemarrhena extractive |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1628790A true CN1628790A (en) | 2005-06-22 |
Family
ID=34846179
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410054146 Pending CN1628790A (en) | 2004-08-31 | 2004-08-31 | Injecta of anemarrhena extractive |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1628790A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229316B (en) * | 2008-01-31 | 2010-08-25 | 广东药学院 | Rhizoma anemarrhenae extrac |
| WO2011026259A1 (en) | 2009-09-07 | 2011-03-10 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Pharmaceutical compositon for anti-thrombotic diseases, its preparation method and use |
| CN101214253B (en) * | 2008-01-07 | 2011-09-21 | 中国人民解放军第二军医大学 | Application of anemarrhenasaponin B-II in preparing antidepressant product |
| CN105330709A (en) * | 2015-06-10 | 2016-02-17 | 中国药科大学 | Method for simultaneously preparing four drug effect components in Rhizoma Anemarrhenae |
| CN105582104A (en) * | 2015-12-30 | 2016-05-18 | 中国中医科学院中医临床基础医学研究所 | Traditional Chinese medicine composition for treating rheumatoid arthritis |
| CN106109416A (en) * | 2016-08-05 | 2016-11-16 | 上海中医药大学 | HHG-001 nanometer liposome and its preparation method and application |
-
2004
- 2004-08-31 CN CN 200410054146 patent/CN1628790A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101214253B (en) * | 2008-01-07 | 2011-09-21 | 中国人民解放军第二军医大学 | Application of anemarrhenasaponin B-II in preparing antidepressant product |
| CN101229316B (en) * | 2008-01-31 | 2010-08-25 | 广东药学院 | Rhizoma anemarrhenae extrac |
| WO2011026259A1 (en) | 2009-09-07 | 2011-03-10 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Pharmaceutical compositon for anti-thrombotic diseases, its preparation method and use |
| CN105330709A (en) * | 2015-06-10 | 2016-02-17 | 中国药科大学 | Method for simultaneously preparing four drug effect components in Rhizoma Anemarrhenae |
| CN105330709B (en) * | 2015-06-10 | 2018-04-03 | 中国药科大学 | Method that is a kind of while preparing four kinds of effective components in the wind-weed |
| CN105582104A (en) * | 2015-12-30 | 2016-05-18 | 中国中医科学院中医临床基础医学研究所 | Traditional Chinese medicine composition for treating rheumatoid arthritis |
| CN106109416A (en) * | 2016-08-05 | 2016-11-16 | 上海中医药大学 | HHG-001 nanometer liposome and its preparation method and application |
| CN106109416B (en) * | 2016-08-05 | 2019-04-09 | 上海中医药大学 | Anemarrhena saponin AIII nano liposomes and its preparation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2021101926A1 (en) | Tryptamine compositions for enhancing neurite outgrowth | |
| CN1754541A (en) | Steroid saponin pharmaceutical composition and its preparation method and uses | |
| CN1526400A (en) | Tubiflorous desert cistanche prepn containing phenethyl alcohol glycoside and its prepn process and use | |
| CN101040915A (en) | Method for preparing a Shuanhuanglian injection and the component detecting method | |
| CN1895302A (en) | Cercopithecoidealin with anti-tumor function, its preparation and use | |
| CN1161389C (en) | Preparing process and use of ginkgo leaf polypentenol and ginkgo leaf extract | |
| CN1628790A (en) | Injecta of anemarrhena extractive | |
| CN1785261A (en) | Hotseason grow corydaline, its preparation method and its medicinal composition | |
| CN101032503A (en) | Effective elements of Chinese traditional medicine combination for curing cardiovascular disease and the quality control method | |
| CN1839932A (en) | Red sage root formulation for venous injection and preparation process thereof | |
| CN1236319A (en) | Pharmaceutical composition containing a mixed extract of phellodondron amurense amurense ruprecht cortex and patrinia scabiosaefolia fish for treatment of hepatitis C | |
| KR20100097517A (en) | A method for isolating and producing highly-concentrated eupatilin and jaceosidine from the extract of artemisia species by using centrifugal partition chromatography | |
| CN1174973C (en) | Schizonepeta lactone and its extraction process and use | |
| CN1543964A (en) | Application in antidepressants preparation of asiatic acid and derivation | |
| CN1186051C (en) | 'Huajuhong' preparation and its preparing process | |
| CN1947736A (en) | Prepn. method and application of injection contg. Erigeron breviscapus | |
| CN1679648A (en) | Mailuoning injection and its preparation and quality control | |
| CN101045077A (en) | Preparating method of oral solid preparation of fenugreek and its use | |
| CN101036647A (en) | Medical composition having anti-inflammatory and anti-infectious function | |
| CN1257710C (en) | Preparation made from effective position of red sage root, and its prepn. method | |
| CN101077341A (en) | Application of aurantio-obtusifolin or its derivates in preparing hypolipidemic drug | |
| KR20150034680A (en) | Coriolus versicolor extracts, methods of isolating biologically-active compounds, and uses thereof | |
| CN100345810C (en) | Novel natural drug effective region of rabdosiaexcisa total diterpene | |
| CN1566136A (en) | Pasqueflower notoginsenosides and extraction method, medicinal uses and pharmaceutical preparation thereof | |
| CN105949044A (en) | Imipramine hydrochloride pharmaceutical composition and medical application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |