CN1424031A - Preparation containing Gingkolactone and its producing process - Google Patents
Preparation containing Gingkolactone and its producing process Download PDFInfo
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- CN1424031A CN1424031A CN02128962.XA CN02128962A CN1424031A CN 1424031 A CN1424031 A CN 1424031A CN 02128962 A CN02128962 A CN 02128962A CN 1424031 A CN1424031 A CN 1424031A
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Abstract
A ginkgolide injection contains ginkgolide A, B and K, and meglucamine as cosolvent. Its advantages are high quality and high stability. A composite medicine containing ginkgolide A, B and K is also disclosed.
Description
Technical field
The invention relates to the preparation of bilobalide, particularly contain the injection of bilobalide, this injection is that active ingredient is prepared from the extract bilobalide of Chinese medicine Folium Ginkgo, belongs to drug world.
Background technology
Bilobalide is to extract the active substance that obtains in the middle of the Chinese herbal medicine Folium Ginkgo, this material can also further be subdivided into ginkalide A on structure, B, C, (bilobalide K is the newfound ginkgolide monomer of inventor to K, separate case application) etc., extracting method for bilobalide has had detailed bibliographical information, for example, people such as Li Xingang have introduced the laboratory extracting method of bilobalide (referring to Li Xingang etc., the laboratory Study on Extraction Method of bilobalide in the Folium Ginkgo, 1998 the 1st phases of Chinese Journal of Pharmaceuticals), Chinese patent literature CN1195665A has introduced the extracting method of bilobalide and has contained the preparation of bilobalide, this method is that Folium Ginkgo is extracted with water boil, with adsorbent extraction filtrate is adsorbed, reuse ethanol desorption, reclaim ethanol, with the crystallization dissolving of separating out, recrystallization, drying makes bilobalide.Chinese patent literature CN1313287A discloses a kind of production technology of bilobalide.The method of separating ginkgolide monomer in the middle of the total extract of bilobalide, a lot of relevant bibliographical informations are also arranged, for example, people such as You Song have introduced the separation of bilobalide in the Folium Ginkgo and structure determination method (referring to You Song etc., the separation of bilobalide and structure determination in the Folium Ginkgo, Chinese pharmaceutical chemistry magazine the 4th phase of nineteen ninety-five).Chinese patent literature CN1287121A discloses the method that is prepared medicine ginkalide A, B by Folium Ginkgo or Folium Ginkgo extractum.Bilobalide can be prepared into it different clinical medicine dosage forms as an effective active substance.The method for preparing injection with bilobalide is open in the middle of Chinese patent literature CN1315175A.Use Folium Ginkgo injections such as Liu Jie treatments acute cerebral infarction obtained certain curative effect [referring to Liu Jie etc., YINXINGYE ZHUSHEYE to acute cerebral infarction after the influence of extremity motor function and TXB_2,6-Keto-PGF_ (1a)].About the pharmacological action of bilobalide, comparatively detailed bibliographical information (referring to Chen Weijun etc., the chemical constitution of bilobalide and Advance on Pharmacological Activities, 1998 the 9th phases of Chinese Pharmaceutical Journal) has been arranged.Although the research for bilobalide and preparation thereof in the middle of the prior art has had a lot of reports,, never stop for the research of bilobalide, particularly, bibliographical information is arranged constantly for the separation of ginkgolide monomer and the research of derivant thereof.Present research for effective monomer in the middle of the bilobalide, structures such as ginkalide A, B, C have been reported, bilobalide is prepared into injection, a very important technical problem is to controllable quality, up to the present, does not still have to find accurately to control the method for the central active constituent content of bilobalide injection product, and, in the middle of the bilobalide injector preparation process, also extremely important for the selection of cosolvent, this directly has influence on the stability and the clinical efficacy of product.The present invention is by the mensuration of active ingredient finger printing in the middle of the bilobalide injection product, the quality of control product, particularly, on the architecture basics of having found bilobalide K, increased a new quality control index, make the suitability for industrialized production of bilobalide injector be achieved, and research by experiment, select ideal cosolvent, thereby can be good at controlling the stability of bilobalide injector product, finished the present invention.
Summary of the invention
The object of the invention provides a kind of preparation that contains bilobalide, particularly bilobalide injector, and the preparation technology of this injection also is provided simultaneously.
Bilobalide injector of the present invention is to be that active ingredient is prepared from highly purified bilobalide, and the weight ratio of injection each component is:
Bilobalide: meglumine: sodium chloride=(2-8): (2-8): (4-12)
The preferred weight proportioning of above-mentioned component is:
Bilobalide: meglumine: sodium chloride=(4-6): (4-6): (6-10)
The preferred weight proportioning of above-mentioned component is:
Bilobalide: meglumine: sodium chloride=5: 5: 8
Wherein can also add 10% an amount of citric acid solution.
Calculate to prepare 1000 milliliters of bilobalide injectors of the present invention, it is composed as follows:
Bilobalide 5.0g
Meglumine 5.0g
NaCl 8.0g
10% citric acid solution is an amount of
The preparation method of bilobalide injector of the present invention is as follows:
Get meglumine and add fresh water for injection 1000ml dissolving, in 120 ℃ of autoclaving 30min, take out the back and be heated to 80 ℃ in gradating material chambers 10,000, add porphyrize and cross the bilobalide of 80 mesh sieves, be heated to dissolving, transfer pH to 8.0 with 10% aqueous citric acid solution after being cooled to room temperature, add NaCl 8.0g, needle-use activated carbon 2.0g, stirring at room absorption 30min, filter carbon removal, add fresh water for injection, serve as a contrast 0.22 μ m filtering with microporous membrane down with aseptic filter plate in 100 grades of sterile workshop to total amount 1000ml, be sub-packed in the 10ml ampoule fusion sealing, quality inspection, lettering, packing.
Prepare the employed ginkgo lactone material of bilobalide injector of the present invention, can extract by known extracting method, perhaps buy by commercially available, still, this raw material need meet the finger printing standard of measuring as follows.
The determining fingerprint pattern of bilobalide:
1. need testing solution preparation
Precision takes by weighing in 105 ℃ of bilobalide sample 30.0mg that are dried to constant weight and puts in the 10.0mL measuring bottle, and with 50% acetone soln dissolving and be diluted to scale, mixing filters through 0.45 μ m microporous filter membrane, promptly.
2. object of reference formulations prepared from solutions
An amount of with the ginkalide B reference substance, add 50% acetone soln and make the solution that every 1mL contains 1mg, promptly.
3. assay method is measured according to high performance liquid chromatography (2000 editions appendix VID of Chinese Pharmacopoeia).
4. chromatographic condition and system suitability test
With the octadecyl silane is filler (Hypersil C
185 μ m 250 * 4.6mm); 30 ℃ of column temperatures; Mobile phase A be water-methanol-oxolane (redistillation) (85: 10: 5, V/V), Mobile phase B be water-methanol-oxolane (redistillation) (35: 60: 5, V/V); Gradient 0～5min, 5%B; 5～55min, 5%B～55%B; 55～56min, 55%～100%B, 100%B keeps 10min; Flow velocity 1.0mL/min; Detect wavelength 220nm, theoretical cam curve should be not less than 7000 in lactone B.
5. assay method is drawn need testing solution 20 μ L injection chromatograph of liquid, promptly.
6. finger printing and technical parameter
Use high effective liquid chromatography for measuring, be 60 minutes writing time.The finger printing of bilobalide is seen Fig. 1, and the finger printing that write down 2 hours is seen Fig. 2.Blank (Fig. 3) is noiseless on inspection.
In this product finger printing, 4 total fingerprint peakses are arranged.Retention time in ginkalide B is 1, all the other total fingerprint peakses are demarcated with relative retention time, in the finger printing, are 1 with the peak area of ginkalide B, calculate the area ratio of each total fingerprint peaks, relative retention time and relative peak area technical parameter see Table 1.
Table 1 bilobalide fingerprint pattern technology parameter
Allowed band when keeping relatively
Total peak relative peak area
Between
1 0.56 0.02
2 0.87 0.56 ±20％
3 1 1 /
4 1.71 1.0 ±20％
In the middle of above-mentioned 4 total peaks, total peak 1 is a ginkalide C, and total peak 2 is ginkalide As, and total peak 3 is ginkalide B, and total peak 4 is bilobalide Ks.
Non-total peak area: the measurement result of test sample, non-total peak area must not be greater than 5%.
Bilobalide in the middle of the bilobalide injector of the present invention comprises ginkalide A, B, C, K, wherein the total content of ginkalide A, B, K is not less than 90%, optimum content is 96.05%, wherein the content of ginkalide B is 50%-65%, the content of ginkalide A is 30-40%, and the content of bilobalide K is 0.5-5%.Calculate by every ml injection product, every milliliter of injection of the present invention contains total bilobalide 2-10 milligram, preferred 4 milligrams.Each monomeric ratio is ginkalide A 35.59%, ginkalide B 58.39%, bilobalide K 2.07% in the middle of described total bilobalide.
Therefore, the invention provides the pharmaceutical composition that contains above-mentioned three kinds of bilobalides, said composition contains:
Ginkalide A 30-40%, ginkalide B 50%-65%, bilobalide K 0.5-5%.
The best proportioning of above-mentioned each component is:
35.59% ginkalide A, 58.39% ginkalide B, 2.07% bilobalide K.
For the ginkgo lactone material purity that makes preparation bilobalide injector of the present invention reaches more than 90.0%, commercially available or homemade ginkgo lactone material (thin layer is differentiated should check out ginkalide A, B, C, K) is dissolved in the ethanol of warm (35-45 degree centigrade), filter, 50 degrees centigrade of reclaim under reduced pressure part ethanol, placement, cooling, crystallize, filter, the residual liquid otherwise processed, getting the crystal of separating out detects, do not reach more than 90% as purity, repeat purification step as stated above, until reaching content requirement.
After ginkgo lactone material reached purity requirement, sample thief 100g added cold ethanol or acetone 100ml, stir, leave standstill sucking filtration, press fingerprint atlas detection method and detect, do not reach the finger printing requirement, repeat above-mentioned treatment step and reach requirement until sample as ginkgo lactone material.That is ginkalide A, B, K and other components in proportions scopes reach the finger printing requirement.
The product of bilobalide injector of the present invention after testing, should meet the requirement of product fingerprint collection of illustrative plates standard.
The determining fingerprint pattern standard of bilobalide injection product of the present invention is as follows:
1. test sample preparation
Precision is measured sample solution 5.0mL and is put in the 10.0ml volumetric flask, adds 6mol/L hydrochloric acid solution 2mL, and the mixing room temperature is placed 3.5hr, centrifugal (3000 rev/mins) 10min, abandoning supernatant, precipitation is with 50% acetone solution, mixing, through 0.45 μ m filtering with microporous membrane promptly.
2. object of reference formulations prepared from solutions
An amount of with the ginkalide B reference substance, add 50% acetone soln and make the solution that every 1mL contains 1mg, promptly.
3. assay method and determining instrument
Measure according to high performance liquid chromatography (2000 editions one appendix VI D of Chinese Pharmacopoeia).
Instrument: Agilent 1100 Series (G1312A Pump, G1313A ALS, G1316A Colcom, G1315A DAD);
Chromatographic condition and system suitability test: with the octadecyl silane is filler, and Hypersil C is used in suggestion
185 μ, 4.6 * 250mm chromatographic column; 30 ℃ of column temperatures; Mobile phase A be water-methanol-oxolane (redistillation) (85: 10: 5, V/V), Mobile phase B be water-methanol-oxolane (redistillation) (35: 60: 5, V/V), 0～5min, 5%B; 5～55min, 5%B～55%B; 55～56 min, 55%～100%B, 100%B keeps 10min; Flow velocity 1.0mL/min detects wavelength 220nm, and theoretical cam curve should be not less than 7000 in lactone B.
Assay method is drawn each 20 μ L of need testing solution and is injected the chromatograph of liquid analysis, promptly.
4. finger printing and technical parameter
(1) finger printing
Adopt high effective liquid chromatography for measuring, be 60 minutes writing time.The finger printing of bilobalide injection is seen Fig. 4, and the finger printing that write down 2 hours is seen Fig. 5.Blank (Fig. 6) is noiseless on inspection.
(2) demarcation of total fingerprint peaks
In the finger printing, 5 total fingerprint peakses should be arranged.Retention time in ginkalide B is 1, and all the other total fingerprint peakses are demarcated with relative retention time.
The chromatographic peak that all occurs in 10 batches of injection is self object of reference as the total peak of injection with ginkalide B respectively, calculates the relative retention time (table 2) at each total peak, with this foundation of demarcating as total fingerprint peaks.The collection of illustrative plates of each batch test sample is seen accompanying drawing (4), figure (5).
Table 2 bilobalide injection relative retention time
123456789 10 is average
Penetrate 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 liquid 1.72 1.72 1.72 1.72 1.72 1.72 1.72 1.71 1.72 1.72 1.72 0.56 0.55 0.56 0.56 0.56 0.55 0.56 0.56 0.56 0.56 0.56 annotates 0.87 0.87 0.87 0.86 0.86 0.87 0.87 0.86 0.87 0.87 0.87
1.75 1.75 1.75 1.75 1.75 1.75 1.75 1.75 1.75 1.75 1.75
The relative retention time at 5 total peaks is in the finger printing:
0.58(1)、0.87
(2)、1(3)
(S)、1.72(4)、1.75
(5)；
In the middle of these 5 total peaks, the 1st, ginkalide A, the 2nd, ginkalide A, the 3rd, ginkalide B, the 5th, bilobalide K is the ginkalide A conversion product and relative retention time is 1.72 peak.
(3) ratio of total fingerprint peaks area
In the finger printing, be 1 with the peak area of ginkalide B, the area ratio of calculating each total peak should meet the regulation of bilobalide injection fingerprint pattern technology parameter (seeing Table 3).
Table 3 bilobalide injection fingerprint pattern technology parameter
Total peak relative retention time relative peak area allowed band
1 0.58 0.03 /
2 0.87 0.53 ±25％
3 1 1 /
4 1.72 0.25 /
5 1.75 4.75 ±20％
The peak area ratio at 5 total peaks is in the finger printing:
0.03 (1), 0.53 ± 25%
(2), 1 (3)
(S), 0.25 (4), 4.75 ± 20%
(5)(4) non-total peak area
The measurement result of test sample, non-total peak area must not be greater than 5%.
The contained bilobalide K of bilobalide injector of the present invention is the newfound a kind of chemical compound of inventor, and this chemical compound separate case is applied for a patent, and its extraction separation method is as follows:
Get bilobalide crude product 3g and put in the apparatus,Soxhlet's, add ethyl acetate (or acetone) 2500ml and refluxed 24 hours, reclaim ethyl acetate to the extractum shape, mix sample with kieselguhr, oven dry, porphyrize, use petroleum ether: ethyl acetate/wet method was loaded on low pressure silicagel column (pressure 0.2-0.3k/cm in 9: 1
2), with petroleum ether: the ethyl acetate gradient increases progressively eluting, when petroleum ether: during ethyl acetate/6: 4,97～121 flow points, through TCL check (toluene: ethyl acetate: acetone: methanol/5: 2.5: 2.5: 0.3,4%NaAc silica gel H plate) occur the bilobalide speckle (with the bilobalide reference substance relatively), it merged concentrate, place the appearance precipitation, mother solution is continued to employ.To precipitate with acetone solution,, can get bilobalide K with preparation type high performance liquid chromatogram purification.122～127 flow points are identical through thin layer inspection (condition is the same) speckle, with its merging, concentrate, and place, and separate out white needle, filter white needle, TCL checks and ginkalide A reference substance R
fValue (0.64) unanimity, ethyl alcohol recrystallization gets the pure product of ginkalide A (GA).Mother liquid obtained and the above-mentioned mother solution of continuing to employ is merged, concentrates a small amount of silica gel mixed sample in back, oven dry, porphyrize, with the chloroform wet method silicagel column (100-140 order silica gel) of packing into, chloroform: third paulownia/9: 1 eluting, the 5-10 flow point, speckle R after the TCL inspection (condition is the same)
fBe worth identically, merge, be concentrated into small size, place Flos Chrysanthemi shape crystallization (R
fValue 0.62) through ethanol repeatedly recrystallization get colourless prismatic crystallization, be reference substance with ginkalide B, check R with ginkalide B through TCL
fBe worth identically, tentatively be defined as ginkalide B (GB).Column chromatography continues to be eluted to the 15-22 flow point, and TCL checks that speckle is identical, merges, and concentrates, place white, needle-shaped crystals, check R with the ginkalide C reference substance through TLC
fBe worth identically, tentatively be defined as ginkalide C (GC).
The conclusive evidence of bilobalide K: white needle (MeoH), mp＞300 ℃ (decomposition) is soluble in alcohol, acetone, is slightly soluble in water.Show slightly faint yellow (place in the air and fade to white).
IRV
max kBr(cm
-1)：
3500，3385，3130，2960，2915，2857，1783，1757，1700，1600，1465，1437，1400，1376，1368，1356，1340，1305，1280，1260，1243，1231，1215，1183，1174，1150，1124，1110，1087，1063，1037，1021，957，750；
ECI-MS(m/z)：405[M-H]
-。
This chemical compound carries out full ownership by physico-chemical analysis and spectroscopic technique in modern age (C-H is relevant, H-H is relevant, HMQC and HMBC spectrum) to the hydrocarbon signal of chemical compound, and the result is as follows.
The hydrocarbon signal ownership and the proton signal of being correlated with (DMSO-d6) position thereof of bilobalide K
13C-NMR
1H-NMR HMBC1 84.00 4.80 (dd) 5.11 (1-OH), (6) 5.53 (2) 2 80.20 6.53 5.47 (dd) 4.83 (1), 5.28 (1-OH) 3 160.33 2.90 (16), 5.53 (2) 4 90.50 6.06 (12) 5 76.39 2.19 (7), 3.83 (1), (10) 6 88.91 6.47 5.00 (d) 2.19 (7), 3.83 (1) 7 35.61 2.208 50.83 1.87 2.87 (8) 9 69.17 10 69.05 6.00 (d) 7.18 (10-OH) 11 74.11 5.00 (10), (12) 12 90.58 6.01 6.06 (S) 1.87 (8) 13 69.18 5.47 (6) 14 25.14 1.91 (16), 5.53 (2) 15 173.43 1.91 (16) 16 18.83 1.91 (d) 17 31.97 1.05 (18-20) 18-20 30.81 1.87 (8)
Annotate:
13C-NMR,
1H-NMR, HMBC refer to carbon-13 nmr spectra, proton nmr spectra, the long-range relevant spectrum of two-dimentional C-H respectively.
Determine that according to above-mentioned wave spectrum feature its chemical structural formula is as follows:
The chemical name of bilobalide K of the present invention is: 1, and 10-dihydroxy-3,14-two dehydrogenation bilobalides (1,10-Dihydroxy-3,14-didehydroginkgolide).Molecular formula C
20H
22O
9, molecular weight is 406.
Bilobalide injector of the present invention has good pharmacologically active, and the result is as follows for its pharmacological experiment study:
1, the general pharmacology of bilobalide is learned research
(1), bilobalide injection 25,50,100mg/kg tail vein injection all do not have obvious influence to mice behavior and coordination exercise, pentobarbital sodium to sub-threshold dose does not also have synergism, shows that three dosage of this medicine are to mice spirit nervous system non-evident effect.
(2), bilobalide injection 5,10, the quiet notes administration of 20mg/kg all do not have obvious influence to blood pressure, heart rate, electrocardio, respiratory frequency and the amplitude of anesthetized dog, shows that three dosage of this medicine are to the anesthetized dog cardiovascular system respiratory system non-evident effect of unifying.
2, the pharmacodynamic study of bilobalide
(1), bilobalide injection 7.5,15.0,30.0mg/kg iV can make the apoplexy scoring of MCAO rat reduce, the MCAO infarction size dwindles, brain water content reduces (P＜0.01), its action intensity and Ginaton injection comparison no significant difference.
(2), bilobalide injection 7.5,15.0,30.0mg/kg iV can make the MDA in the cerebral tissue of focal cerebral ischemia rat reperfusion injury, the LA equal size reduces (P＜0.01), shows that cerebral tissue hypoxic-ischemic and peroxidating degree are subjected to obvious inhibition; SOD and GSH content increase (P＜0.01) simultaneously, have reflected that medicine has raising to antioxidant ability of organism and the ability of removing free radical.
(3), bilobalide injection 7.5,15.0,30.0mg/kg iV can obviously protect the cerebral tissue structure of focal cerebral ischemia rat reperfusion injury; karyopycnosis, the karyolysis degree of cerebral cortex pyramidal cell and brain essence neurocyte obviously alleviate than ischemic control group, and softening kitchen range reduces.
(4), bilobalide injection 7.5,15.0,30.0mg/kgiV compare with model group that the EEG that can make diffusivity global brain ischemia rat recovers normal time and (P＜0.01) is obviously shortened in righting reflex recovery time, make the azovan blue permeability significantly reduce (P＜0.01), its effect does not relatively have statistical significant difference with the Ginaton injection.
(5), bilobalide injection can obviously increase the survival number of acute imperfection cerebral ischemia mice.Press the ED that the Bliss method is calculated bilobalide injection iV
50Be 21.3 (17.6-25.6) mg/kg; The ED of bilobalide ig
50Be 29.2 (16.5-51.7) mg/kg; The ED of Ginaton iV
50Count 12.2 (6.3-23.3) mg/kg, ED with Semen Ginkgo extrac
50Value compares zero difference through between t check group.
(6), bilobalide injection iV10min promptly begins onset, about peak time 20min, the persistent period can reach more than the 90min; And this medicine ig onset time 20-30min, about peak time 45min, the persistent period can reach more than the 120min.The effect of same dosage group iV is better than ig.
(7) but, bilobalide injection 12.0,24.0, the time-to-live (P＜0.01) of 48.0mg/kg iV significant prolongation mice under anaerobic condition.
(8), bilobalide injection 2.5,5.0,10.0mg/kg iV all do not have obvious influence (P＞0.05) to anesthetized dog CBF, CVR, SBP, DBP, MBP and HR, shows that three dosage of this medicine do not influence cerebral blood flow, cerebral vascular resistance, blood pressure and the heart rate of anesthetized dog.
(9), bilobalide injection 0.99,1.96,3.80 μ g/ml induce the rabbit extracorporeal platelet aggregation that obvious inhibitory action (P＜0.01) is all arranged to PAF, and obvious depolymerisation (P＜0.01) are arranged.To ADP induced platelet aggregation weak (P＜0.05 or P＜0.01), there is not obvious depolymerisation (P＞0.05).This medicine iV0.75,1.50,3.0mg/kg all have obvious inhibitory action (P＜0.01) to platelet aggregation in the inductive rabbit body of PAF, and middle and high dosage group also has obvious depolymerisation (P＜0.01).The rabbit platelet number there is not obvious effect (P＞0.05).
(10), bilobalide injection iV1.50,3.0mg/kg can obviously reduce rabbit erythrocyte hematocrit (P＜0.01) and whole blood viscosity (ratio) especially to whole blood viscosity (ratio) effect more obvious (P＜0.01) of high shearing.
(11), it is moving that bilobalide injection iv 0.75,1.50,3.0mg/kg can obviously suppress rabbit---the thrombosis (P＜0.01) of vein bypass.
(12), bilobalide injection iv 3,6,12mg/kg are moving to rat---and vein bypass thrombosis has obvious thrombolytic effect (P＜0.01).
(13), bilobalide injection iv6,12,24mg/kg can obviously prolong clotting time of mice (P＜0.01).
(14), obvious prolong rats clotting time during 10min-20min behind the bilobalide injection iv6mg/kg, during the peak about 12min, can make rat cruor time extending about 32%, 45min is effective behind the ig50mg/kg, continue 120mm, during the peak about 69min, rat cruor time extending about 34%.
(15), just can obviously suppress PAF behind the bilobalide injection iv 1.5mg/kg medicine immediately induces and exempts from the body platelet aggregation and obvious depolymerisation (P＜0.01) is arranged, effect continues 30min, be 5min during its peak, platelet aggregation inhibition rate can reach about 80%, and depolymerization in 1 minute is about 81%.30min can obviously suppress PAF and induces in the rabbit body platelet aggregation and obvious depolymerisation (P＜0.01) is arranged behind the bilobalide ig25mg/kg medicine, continue 60min, be about 42min during its peak, the maximum gathering of platelet suppression ratio is about 38%, and the depolymerization rate was about 32% in 1 minute.
(16), bilobalide injection iV prolongs the IC of clotting time of mice
50Be 6.42mg/kg, bilobalide ig prolongs the IC of clotting time of mice
50Be 29.41mg/kg.Bilobalide injection iv is to the IC of platelet aggregation in tame rabbit platelet and the PAF inductor
50Be 2.32mg/kg, bilobalide ig is to the IC of platelet aggregation in tame rabbit platelet and the PAF inductor
50Be 87.72mg/kg.
Above-mentioned experiment shows that bilobalide injection of the present invention is safe and effective, can be used for the prevention and the clinical treatment of cerebral infarction.
The advantage of bilobalide injection of the present invention is that constant product quality is controlled, according to the finger printing of injection product, can control the composition of product, particularly increased this controlling index of bilobalide K, cooperate indexs such as other ginkalide As, B, C, make that stablizing of product is more controlled.
Bilobalide injector of the present invention has stability and better curative effect preferably, also because the preparation process of bilobalide injector of the present invention, the selection of cosolvent has significant effects to the preparation and the stability of preparation, in the research process of the present invention, the investigation of system (1) propylene glycol, PEG
400, ethanol, (2) propylene glycol, PVP, ethanol, (3) propylene glycol, sorbitol, ethanol, (4) meglumine, (5) meglumine, glucose, (6) carbamide, (7) propylene glycol, tween 80, ethanol, (8) propylene glycol, ethanol, (9) sodium hydroxide, (10) Pluronic F-68, (11) Pluronic F-68, propylene glycol, (12) fabaceous lecithin, PluronicF-68, propylene glycol, glycerol, PEG
400, ethanol, (13) HP-β CD, (14) propylene glycol, PEG
400, phosphate buffer, (15) propylene glycol, PEG
400, multiple cosolvent such as phosphate solution.By a large amount of tests, find that above-mentioned pharmaceutic adjuvant all help solubilization, wherein with the meglumine best results.This cosolvent all has good hydrotropy effect to the combination (for example: ginkalide A, B, ginkalide A, B, C, ginkalide B, K, ginkalide A, B, C, K, ginkalide B, C, K, ginkalide A, C etc.) of other components of bilobalide.
Description of drawings
Fig. 1 bilobalide finger printing
2 hours finger printing of Fig. 2 bilobalide record
Fig. 3 bilobalide finger printing blank
Fig. 4 is the bilobalide injection finger printing
2 hours finger printing of Fig. 5 bilobalide injection record
Fig. 6 bilobalide injection finger printing blank
The specific embodimentPrescription:
Bilobalide 5.0g
Meglumine 5.0g
NaCl 8.0g
10% citric acid solution is an amount of
Method for making: get meglumine and add fresh water for injection 1000ml dissolving, in 120 ℃ of autoclaving 30min, take out the back and be heated to 80 ℃ in gradating material chambers 10,000, add porphyrize and cross the bilobalide (bilobalide meets quality standard of the present invention) of 80 mesh sieves, be heated to dissolving, transfer pH to 8.0 with 10% aqueous citric acid solution after being cooled to room temperature, add NaCl 8.0g, needle-use activated carbon 2.0g, stirring at room absorption 30min, filter carbon removal, add fresh water for injection, serve as a contrast 0.22 μ m filtering with microporous membrane down with aseptic filter plate in 100 grades of sterile workshop to total amount 1000ml, be sub-packed in the 10ml ampoule fusion sealing, quality inspection, lettering, packing.
This product is the sterile water solution of bilobalide.Every ml bilobalide-containing A, B, K total amount must not be lower than 4.0mg.
Claims (19)
1. a pharmaceutical composition is characterized in that said composition contains ginkalide A 30-40%, ginkalide B 50%-65%, bilobalide K 0.5-5%.
2. according to the pharmaceutical composition of claim 1, it is characterized in that said composition contains 35.59% ginkalide A, 58.39% ginkalide B, 2.07% bilobalide K.
3. bilobalide injector, the primary raw material that it is characterized in that this injection is total bilobalide, this raw material bilobalide-containing A, B, K total amount are greater than 90%.
4. according to the injection of claim 3, it is characterized in that the total bilobalide of this injection primary raw material contains ginkalide A 30-40%, ginkalide B 50%-65%, bilobalide K 0.5-5%.
5. according to the injection of claim 4, it is characterized in that every milliliter of injection contains total bilobalide 2-10 milligram.
6. according to the injection of claim 5, it is characterized in that every milliliter of injection contains 4 milligrams of total bilobalides.
7. according to the injection of claim 6, it is characterized in that each monomeric ratio is 35.59% ginkalide A, 58.39% ginkalide B, 2.07% bilobalide K in the injection institute bilobalide-containing.
8. according to the injection of claim 3, it is characterized in that the total bilobalide of this injection primary raw material adopts its finger printing of high effective liquid chromatography for measuring, be 60 minutes writing time, get 4 total peaks, retention time 1 with ginkalide B is self object of reference, the relative retention time at these 4 total peaks is 0.56,0.87,1,1.71, is 1 with the peak area of ginkalide B, and the peak area ratio at these 4 total peaks is 0.02,0.56 ± 20%, 1,1.0 ± 20%.
9. according to the injection of claim 3, it is characterized in that this injection adopts its finger printing of high effective liquid chromatography for measuring, 60 minutes writing times, get 5 total peaks, retention time 1 with ginkalide B is self object of reference, the relative retention time at these 5 total peaks is 0.58,0.87,1,1.72,1.75, is 1 with the peak area of ginkalide B, and the peak area ratio at these 5 total peaks is 0.03,0.53 ± 25%, 1,0.25,4.75 ± 20%.
10. according to the injection of claim 9, it is characterized in that the central non-total peak area of this injection finger printing is less than 5%.
11. a bilobalide injector is characterized in that containing meglumine in the middle of this injection.
12. according to the injection of claim 11, it is characterized in that this injection also contains sodium chloride, the each component weight ratio is: bilobalide: meglumine: sodium chloride=(2-8): (2-8): (4-12).
13., it is characterized in that the each component weight ratio of this injection is: bilobalide: meglumine according to the injection of claim 12: sodium chloride=(4-6): (4-6): (6-10).
14., it is characterized in that the each component weight ratio of this injection is: bilobalide: meglumine: sodium chloride=5: 5: 8 according to the injection of claim 13.
15., it is characterized in that also adding 10% an amount of citric acid solution in the middle of the component of this injection according to the injection of claim 14.
16. an injection, it contains the bilobalide K for the treatment of effective dose to it is characterized in that this.
17. according to the injection of claim 16, it is characterized in that it contain 0.5-5% bilobalide K.
18., it is characterized in that it contains meglumine according to the injection of claim 16.
19. the preparation method of a bilobalide injector is characterized in that the prescription of this injection and method for making are as follows:
Bilobalide 5.0g
Meglumine 5.0g
NaCl 8.0g
10% citric acid solution is an amount of
Get meglumine and add fresh water for injection 1000ml dissolving,, take out the back and is heated to 80 ℃ in proportioning room in 120 ℃ of autoclavings 30 minutes, add porphyrize and cross the bilobalide of 80 mesh sieves, be heated to dissolving, be cooled to after the room temperature aqueous citric acid solution and transfer pH to 8.0, add NaCl 8.0g with 10%, needle-use activated carbon 2.0g, carbon removal is filtered in stirring at room absorption 30 minutes, adds fresh water for injection to total amount 1000ml, serve as a contrast 0.22 μ m filtering with microporous membrane, packing in sterile workshop down with aseptic filter plate.
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| CNB200610086424XA CN100401059C (en) | 2002-08-23 | 2002-08-23 | Fingerprint atlas quality investigating method of ginkgo lactone material |
| CN02128962.XA CN1279903C (en) | 2002-08-23 | 2002-08-23 | Preparation containing Gingkolactone and its producing process |
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| CN02128962.XA CN1279903C (en) | 2002-08-23 | 2002-08-23 | Preparation containing Gingkolactone and its producing process |
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| CNB200610086424XA Division CN100401059C (en) | 2002-08-23 | 2002-08-23 | Fingerprint atlas quality investigating method of ginkgo lactone material |
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| CN1424031A true CN1424031A (en) | 2003-06-18 |
| CN1279903C CN1279903C (en) | 2006-10-18 |
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| CNB200610086424XA Expired - Lifetime CN100401059C (en) | 2002-08-23 | 2002-08-23 | Fingerprint atlas quality investigating method of ginkgo lactone material |
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Also Published As
| Publication number | Publication date |
|---|---|
| CN1869683A (en) | 2006-11-29 |
| CN100401059C (en) | 2008-07-09 |
| CN1279903C (en) | 2006-10-18 |
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