CN1907291A - Injectio for nanometer notoginsenoside and its preparing method - Google Patents
Injectio for nanometer notoginsenoside and its preparing method Download PDFInfo
- Publication number
- CN1907291A CN1907291A CN 200510089090 CN200510089090A CN1907291A CN 1907291 A CN1907291 A CN 1907291A CN 200510089090 CN200510089090 CN 200510089090 CN 200510089090 A CN200510089090 A CN 200510089090A CN 1907291 A CN1907291 A CN 1907291A
- Authority
- CN
- China
- Prior art keywords
- aescine
- preparation
- nanometer
- add
- schemes
- 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
Abstract
The invention discloses a nanometer aescin injection and preparing method, which is characterized by the following: extracting from effective position from aescin to prepare nanometer aescin with medical carrier; blending with drug auxiliary material to prepare water injection, transfusion agent and powder injection; controlling the content of beta-aescin at 50-85%.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicine pharmacy, be specifically related to a kind of nanometer aescine ejection preparation and preparation method thereof.
Background technology
Semen Aesculi is the fruits and seeds of Hippocastanaceae plant Aesculus hippocastanum L., Aesculus turbinata B1, Chinese Heavenly Teacher chestnut, and its invalid components is fatty oil, starch, fiber etc., and fatty oil is mainly oleic acid and stearic glyceride; Its effective ingredient is an aescine, is to belong to triterpene saponin compound, and it has antiinflammatory, exudation recovers capillary permeability, improves intravenous tension, improve blood circulation, have expansion artery concurrently, coronary artery dilating, increase hypoxia-bearing capability, promote effects such as brain function recovery, clinical practice mainly is its sodium salt, i.e. aescin for injection, but in clinical practice, a large amount of serious adverse reactions appears: 1. cause hepatic injury; 2. cause anaphylaxis blister type tissue necrosis; 3. hyperamization is urinated; 4. cause the systemic anaphylaxis erythra; 5. cause anaphylactic shock; 6. cause acute renal failure; 7. cause bradycardia; 8. cause the vein infringement.[" Tianjin pharmacy ", Tianjin Pharmacy, in April, 2004, the 16th the 2nd phase of volume, 48-50] therefore, the medical scientific research research worker should be carried out preparation research with aescine, develops rapid, the safe aescine preparation of a kind of onset; Seven leaf soaps mainly contain two kinds of isomers of α, β, wherein β-aescine is a main active, β-aescine is based on aescine A, B, but β-aescine aqueous solution is in the time of 100 ℃, acetyl group can take place for C-21, C-22, C-21, C-28 hydroxyl shifts, change into α-aescine, make active reduction [" World Science technology-Chinese medicine modernization " the 3rd phase in 2004 of aescine effective site, 45-46], therefore, in the process of extracting purification aescine effective site, should note temperature controlling; Consult document and patent, in the process of extracting aescine, adopt alcohol extraction more, temperature is not extracted in control, and invalid components oils composition is kept, and has strengthened the difficulty of purge process; In purge process, there is report to adopt organic solvent extractionprocess, has report to adopt D101 type or NKA-9 type macroporous adsorptive resins to carry out ethanol elution, in handling the process that reclaims organic solvent, do not have fine control temperature yet, therefore can cause the content of β-aescine in the aescine to reduce.
Liposome is the middle made superminiature spheroid carrier preparation of thin film that drug encapsulation is formed in the lipoids bilayer.The material that constitutes lipid bilayer (carrier) mainly contains phospholipid (lecithin, cephalin, fabaceous lecithin) and cholesterol etc., because existing hydrophilic group has hydrophobic group again in their molecular structure, when constituting spherical class Lipid Bilayer Structure, the structure that its hydrophobic group constitutes is relatively got up drug encapsulation.Liposome has the following advantages as the carrier of medicine: (1) this carrier can be protected enveloped; (2) control drug release effectively; (3), can control medicine in in-house distribution and the clearance rate in blood by changing liposome size and electric charge; (4) change certain physical factor, the permeability that temperature that for example changes the partial pH of medication, diseased region etc. can obviously change liposome membrane impels liposome optionally to discharge medicine; (5) after liposome enters in the body mainly by reticuloendothelial system phagocytic, can activate the autoimmune function of body, and drug main will be accumulated in histoorgans such as brain, liver and bone marrow, thus improve the therapeutic index of medicine, reduce the therapeutic dose of medicine and reduce drug toxicity; (6) liposome itself is to human body avirulence and immunosuppressive action.Therefore, liposome especially demonstrates obvious superiority in the brain diseases treatment at numerous disease.
Consult document and patent, do not retrieve the data of nanometer aescine ejection preparation.
Summary of the invention
For these reasons, we are by a large amount of experimentatioies, Semen Aesculi is adopted ultrasonic water extraction earlier, the control temperature is no more than 50 ℃, stand at low temperature is removed the oils invalid components, adopt ethanol ultrasonic extraction again, the control temperature is no more than 50 ℃, in purge process, we adopt AB-8 type macroporous adsorptive resins to be prepared, and all concentration processs of the present invention adopt rotation wiped film vaporization instrument to carry out, the control temperature is no more than 50 ℃, in whole extraction purge process, better controlled temperature, prevent that active component β-aescine is converted into invalid components α-aescine in the aescine effective site.
Aescine effective site be will obtain and chemical property and analysis of physical carried out, mix with preferred solvent, lipid according to its character, control temperature well, be prepared into the nanometer aescine, be mixed with into aqueous injection, infusion solution, injectable powder with pharmaceutic adjuvant, pharmacodynamic study, nanometer aescine ejection preparation of the present invention has characteristics safe, that pharmacological action is good.
The purpose that the present invention makes nanometer formulation with aescine is to solve the reduction untoward reaction, reaches rapid-action and keeps good blood drug level.
The purpose of this invention is to provide a kind of nanometer aescine ejection preparation;
Another object of the present invention provides the method for preparing nanometer aescine ejection preparation.
The present invention is achieved through the following technical solutions:
One, process recipes
(1) extraction of aescine effective site:
Get Semen Aesculi, pulverize, put into the supersound extraction jar, 25 ℃-50 ℃ of temperature are extracted in control, extract each 5-15 minute 2-4 time, residue is standby, merge extractive liquid, concentrates 40 ℃-50 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.1-0.2 atmospheric pressure, relative density is 1.15-1.25 when being concentrated to 40 ℃-50 ℃, and concentrated solution was left standstill 24-48 hour at 0 ℃-4 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make to contain the alcohol amount, put into the supersound extraction jar for 50%-70%, 25 ℃-50 ℃ of control temperature, extract 2-4 time, each 15-45 minute, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 40 ℃-50 ℃ of control thickening temperatures, vacuum is 0.1-0.2 atmospheric pressure, obtains concentrated solution; Get macroporous adsorptive resins on the above-mentioned concentrated solution, with the distilled water eluting of 4-8 column volume doubly, eluent discards, the ethanol elution of reuse 60%-90% earlier, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 40 ℃-50 ℃ of control thickening temperatures, vacuum is 0.1-0.2 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site;
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 8-12 weight portion, lipid 48-142 weight portion, emulsifying agent 32-58 weight portion;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 70-90 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 70-90 ℃ of logical condition of nitrogen gas, spare 5-7 time in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, preparation cost invention nanometer aescine hydro-acupuncture preparation;
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, preparation cost invention nanometer aescine infusion preparation;
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and preparation cost invention nanometer aescine freeze-dried powder.
Lipid in the preparation method of nanometer aescine ejection preparation is a stearic acid.
Emulsifying agent is a soybean lecithin in the preparation method of nanometer aescine ejection preparation.
Two. check and analysis
1. β-aescine (aescine A meter) assay in the aescine
Method according to the Pharmacopoeia of the People's Republic of China (2005 one, 206 pages) [assay] is carried out check and analysis, and experimental result sees Table 1:
2. β-aescine (aescine A meter) assay in the aescine preparation
Method according to the Pharmacopoeia of the People's Republic of China (2005 one, 206 pages) [assay] is carried out check and analysis, and experimental result sees Table 1:
Table 1 content relatively
| Group | β-aescine (aescine A meter) content |
| Otoginsenoside active component % aescin for injection mg/g crude drug nanometer otoginsenoside liquid drugs injection mg/g crude drug nanometer otoginsenoside infusion solution mg/g crude drug of the present invention | 50-85 8.4 14.2 13.9 |
| Nanometer aescine injectable powder mg/g crude drug | 14.0 |
Conclusion: show that by above-mentioned experiment technology of the present invention has practical significance.
Three. optimization experiment
Preferred the foundation: " method among Chinese pharmacopoeia version appendix in 2005 the XIX E " microcapsule, microsphere and Liposomal formulation guideline ".H-7000 type transmission electron microscope instrument (Japanese Hitachi company); Zetamaster photon correlation spectrometer (Britain Malvern company); LC-10A high performance liquid chromatograph (day island proper Tianjin); TGL-18G type table model high speed centrifuge.
Experimental technique: get aescine effective site of the present invention and different lipids: glyceryl tristearate, tripalmitin, trilaurin, three climb over a wall acid glyceride, monostearate, stearic acid, different emulsifiers soybean lecithin, Ovum Gallus domesticus Flavus lecithin, polysorbate, poloxamer carry out the Nano medication preparation, obtain the nanometer aescine and experimentize;
Scheme 1: glyceryl tristearate, soybean lecithin;
Scheme 2: tripalmitin, soybean lecithin;
Scheme 3: trilaurin, soybean lecithin;
Scheme climb over a wall at 4: three acid glyceride, soybean lecithin;
Scheme 5: monostearate, soybean lecithin;
Scheme 6: stearic acid, soybean lecithin;
Scheme 7: glyceryl tristearate, Ovum Gallus domesticus Flavus lecithin;
Scheme 8: tripalmitin, Ovum Gallus domesticus Flavus lecithin;
Scheme 9: trilaurin, Ovum Gallus domesticus Flavus lecithin;
Scheme climb over a wall at 10: three acid glyceride, Ovum Gallus domesticus Flavus lecithin;
Scheme 11: monostearate, Ovum Gallus domesticus Flavus lecithin;
Scheme 12: stearic acid, Ovum Gallus domesticus Flavus lecithin;
Scheme 13: glyceryl tristearate, polysorbate;
Scheme 14: tripalmitin, polysorbate;
Scheme 15: trilaurin, polysorbate;
Scheme climb over a wall at 16: three acid glyceride, polysorbate;
Scheme 17: monostearate, polysorbate;
Scheme 18: stearic acid, polysorbate;
Scheme 19: glyceryl tristearate, poloxamer;
Scheme 20: tripalmitin, poloxamer;
Scheme 21: trilaurin, poloxamer;
Scheme climb over a wall at 22: three acid glyceride, poloxamer;
Scheme 23: monostearate, poloxamer;
Scheme 24: stearic acid, poloxamer;
1. morphologic observation and particle diameter
Above-mentioned Nano medication is carried out morphologic observation under transmission electron microscope, observation index is: be which kind of state, whether size is even, whether the surface smooth, have or not adhesion, gets 1000-2000 Nano medication respectively and measure size, normal distribution whether, and experimental result sees Table 2:
Different group forms of table 2 and size ratio are
| Group | State | Whether size is even | Whether the surface is smooth | Have or not adhesion | Minimum grain size nm | Maximum particle diameter nm | Mean diameter nm | Normal distribution whether |
| Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10 schemes 11 schemes 12 schemes 13 schemes 14 schemes 15 schemes 16 schemes 17 schemes 18 schemes 19 schemes 20 schemes 21 schemes 22 schemes 23 | The a small amount of spherosome spherosome of the spherosome spherosome spherosome spherosome spherosome spherosome a small amount of spherosome of a small amount of spherosome of a small amount of spherosome of a small amount of spherosome of irregular irregular a small amount of spherosome spherosome spherosome spherosome spherosome | Whether no whether no no no | No no whether no no whether no no no | Have or not do not have not have do not have do not have have no have no | 52 79 57 43 41 10 109 127 85 74 65 73 235 241 148 157 89 96 42 44 79 86 85 | 208 304 186 142 199 120 385 571 204 201 186 285 732 741 547 209 164 301 159 158 214 189 157 | 148±18 201±32 124±17 98±7 133±24 65±10 287±46 421±39 142±42 153±34 128±27 207±32 587±103 652±157 406±99 186±48 128±30 207±46 109±14 132±28 189±40 144±27 122±23 | Whether no no whether no whether no no |
| Scheme 24 | Spherosome | Not | Be | Not | 87 | 302 | 248±101 | Not |
2. envelop rate and drug loading are measured
Adopt above-mentioned check and analysis method that the Nano medication of different schemes is carried out the aescine assay, and with following formula computational envelope rate and drug loading:
Envelop rate (%)=(dosage-free drug amount)/dosage * 100%;
Weight * 100% of drug loading (%)=(dosage-free drug amount)/Nano medication.
Experimental result: see Table 3.
Table 3 envelop rate and drug loading are relatively
| Group | Envelop rate | Drug loading |
| ％ | ％ | |
| Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10 schemes 11 schemes 12 schemes 13 schemes 14 schemes 15 schemes 16 schemes 17 schemes 18 schemes 19 schemes 20 schemes 21 schemes 22 schemes 23 | 80.1 74.3 79.2 65.3 63.2 92.0 54.3 57.9 58.9 67.3 72.3 74.6 84.3 49.6 57.6 59.1 68.7 78.3 80.2 57.6 65.3 66.9 78.9 | 7.2 6.3 6.8 6.4 10.2 10.1 7.8 7.9 9.8 4.5 5.2 6.3 5.1 4.3 3.8 3.7 5.8 5.2 6.8 8.7 8.2 8.7 4.9 |
| Scheme 24 | 77.2 | 7.3 |
3. study on the stability
The different schemes Nano medication is placed in the bottle sealing respectively.In the environment of refrigerator (3-5 ℃), room temperature (20-25 ℃) and 37 ℃ (RH75%), place, observe outward appearance, size, redispersibility of Nano medication etc. March.The results are shown in Table 4.
Table 4 stability experiment relatively
| Group | 3-5℃ | 3-5℃ | 20-25℃ | 20-25℃ | 37℃(RH75％) | 37℃(RH75％) |
| Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10 schemes 11 schemes 12 schemes 13 | Particle diameter (nm) 157 ± 24 211 ± 27 128 ± 18 102 ± 9 135 ± 27 66 ± 10 294 ± 48 428 ± 41 144 ± 40 155 ± 33 139 ± 29 214 ± 30 595 ± 104 | The white white variable color variable color of variable color variable color of colours white | Particle diameter (nm) 178 ± 63 237 ± 39 138 ± 29 119 ± 15 139 ± 29 67 ± 11 308 ± 52 429 ± 40 148 ± 49 159 ± 39 147 ± 34 219 ± 42 599 ± 102 | The white variable color variable color white of color variable color variable color variable color variable color | Particle diameter (nm) 187 ± 57 258 ± 58 144 ± 39 128 ± 48 143 ± 34 68 ± 11 324 ± 67 437 ± 49 157 ± 51 173 ± 48 152 ± 28 228 ± 58 607 ± 109 | The white variable color variable color of color variable color variable color white variable color electrochromic variable look variable color variable color variable color |
| Scheme 14 schemes 15 schemes 16 schemes 17 schemes 18 schemes 19 schemes 20 schemes 21 schemes 22 schemes 23 | 667±159 418±102 197±49 131±33 211±45 111±17 137±29 197±48 158±30 131±29 | The white colourless variable color of the colourless white of colourless white variable color variable color | 672±168 427±93 203±53 139±36 219±49 125±24 148±34 203±43 167±37 139±33 | The variable color of the colourless white canescence canescence white of white variable color variable color variable color | 684±157 434±107 212±58 147±47 237±49 139±31 152±39 214±54 178±41 148±47 | Variable color variable color electrochromic variable look white variable color variable color canescence variable color variable color |
| Scheme 24 | 257±106 | Canescence | 266±114 | Variable color | 278±115 | Variable color |
4. blood brain barrier research
Get new zealand rabbit, the about 2.5kg of body weight, male and female are not limit.Intravenous administration, 20 minutes extraction 0.5ml cerebrospinal fluid experimentize after administration.After getting cerebrospinal fluid 0.25ml solubilizer suspendible 15min, leaving standstill 15min, with the centrifugal 1min of 10000r/min, get supernatant 20 μ l sample introductions, is index determining with the aescine, and it is standard index with scheme 6 that experimental result sees Table 5:(, promptly 100%)
The different group cerebrospinal fluid of table 5 Chinese medicine concentration
| Group | Cerebrospinal fluid Chinese medicine concentration (%) |
| Scheme 1 scheme 2 schemes 3 schemes 4 schemes 5 schemes 6 schemes 7 schemes 8 schemes 9 schemes 10 schemes 11 schemes 12 schemes 13 schemes 14 schemes 15 schemes 16 schemes 17 schemes 18 schemes 19 schemes 20 schemes 21 schemes 22 schemes 23 | 85.3 88.3 71.2 84.8 82.3 100 77.3 74.3 77.2 84.2 86.1 87.9 88.3 90.4 74.3 78.5 79.6 84.3 85.6 84.3 72.9 76.8 77.3 |
| Scheme 24 | 80.1 |
Experiment conclusion: by above-mentioned optimization experiment, we determine that lipid is a stearic acid, and emulsifying agent is a soybean lecithin; Show that by above-mentioned experiment as seen Nano medication of the present invention is spherosome, size is more even, smooth surface, no adhesion has been measured 1500 according to the light micrograph of Nano medication, mean diameter is 65 ± 10nm, maximum particle diameter is 120nm, and minimum grain size is 10nm, and particle size distribution meets normal distribution law, envelop rate is 92.0%, drug loading is 10.1%, and good stability is made good use of by blood brain barrier.
Four. toxicological experiment
Experiment 1
Acute toxicity testing
Experiment medicine: nanometer aescine ejection preparation of the present invention (Tianzhijiao Medication Development Co., Ltd., Guangdong's laboratory provides); Commercially available aescin for injection (Yantai Green Leaf Pharmaceutical Co. Ltd)
Laboratory animal: healthy mice 18-22g,, male and female half and half;
Experimental technique: get mice, be divided into commercially available aescin for injection group, this patent nanometer aescine injection group, intraperitoneal administration, be converted into the mice dosage according to body surface area, administration every day 3 times, continuous 7 days, observe the dead mouse situation, record data calculate LD by the improvement karber's method
50The value experimental result sees Table 6:
Table 6 mice medication LD
50Value relatively
| Group | Number of animals only | LD 50 mg/kg |
| Commercially available aescin for injection group nanometer aescine of the present invention aqueous injection nanometer aescine of the present invention infusion solution | 20 20 20 | 2.25 14.94 15.12 |
| Nanometer aescine injectable powder of the present invention | 20 | 15.21 |
Experiment 2
Hemolytic is investigated
Experiment medicine: nanometer aescine ejection preparation of the present invention (Tianzhijiao Medication Development Co., Ltd., Guangdong's laboratory provides); Commercially available aescin for injection (Yantai Green Leaf Pharmaceutical Co. Ltd)
Experimental technique: the preparation of 2% red blood cell suspension: get rabbit ear edge vein and get blood 10-20ml, put into the conical flask that fills bead, Fibrinogen is removed in jolting 10 minutes, makes to become to take off fine blood.Add the normal saline solution of 10 times of amounts, shake up, centrifugal, remove supernatant, sedimentary erythrocyte reuse normal saline solution washing 2-3 time, when supernatant does not take on a red color till.It is 2% suspension that the erythrocyte of gained is made into concentration with normal saline, promptly.
Test method: get 6 in test tube, add 2% red blood cell suspension and normal saline solution successively by the proportional quantity in the table, mixing was placed 30 minutes in 37 ℃ of calorstats, added not commensurability medicinal liquid (is blank with the 6th pipe) respectively, after shaking up, put in 37 ℃ of calorstats, beginning was observed 1 time every 15 minutes, after 1 hour, observed 1 time every 1 hour, observed altogether 2 hours.The results are shown in Table 7.
Each group proportional quantity of table 7
| The test tube numbering | 2% red blood cell suspension (ml) | Normal saline solution (ml) | Medicinal liquid (ml) |
| 1 2 3 4 5 | 2.5 2.5 2.5 2.5 2.5 | 2.0 2.1 2.2 2.3 2.4 | 0.5 0.4 0.3 0.2 0.1 |
| 6 | 2.5 | 2.5 | 0 |
The result: be as the criterion with the 3rd test tube, each Guan Junwei dyes redness, and microscopically is observed and do not seen that erythrocyte fragmentation is arranged, and not haemolysis of this product is described, safety is good.
Experiment 3
Blood vessel irritation is investigated
Experimental technique: get 9 of healthy big ear rabbit, animal is fixed in the rabbit hutch, with ethanol with skin degerming after, in right side auricular vein place injection nanometer aescine ejection preparation 5mg/kg, the grade of injecting same volume in the opposite side corresponding position is oozed G/W in contrast.Inject every day 1 time, continuous 3 times, observe the reaction of rabbit ear edge vein.Behind last administration 24h,, take off two ears, soak,, dissect and take out vein, do the tissue slice inspection, observe the reaction of injection site apart from injection site 1-4cm place with 10% formalin with the rabbit sacrificed by exsanguination.
The result: in the process of the test, place, perusal two ear vein injection site, swollen, the heat of show etc. does not stimulate performance.Show: administration group section position tissue morphology no significant difference, do not see that the pathomorphology due to this product toxicity changes (blood vessel structure is normal, no endothelial cell damage, no thrombosis formation and the variation of other pathologic).
Conclusion: show that by above-mentioned experiment nanometer aescine ejection preparation of the present invention is better than commercially available aescin for injection safety.
Four. pharmacological evaluation
Reagent and animal: aescin for injection (Yantai Green Leaf Pharmaceutical Co. Ltd); Nanometer aescine ejection preparation of the present invention (by Tianzhijiao Medication Development Co., Ltd., Guangdong's prepared in laboratory); Collagenase, U.S. Sigma company produces, new zealand rabbit, the about 2.5kg of body weight; Big ear rabbit, the about 2.5kg of body weight; The SD rat, body weight 180-220g; Kunming mouse, body weight 18-22g.
Experiment 1
Xylol causes the influence of mice auricle swelling
Get 50 of Kunming mouses, 18-22g cuts off the hair around the auris dextra, is divided into five groups at random, i.e. blank group, aescin for injection positive controls, nanometer aescine ejection preparation group of the present invention.(dosage is 2.25mg/kg to be subjected to reagent liquid with the micropipettor absorption, injectable powder dilutes with water for injection) be applied to auris dextra auricle both sides, three times on the one, successive administration five days, each dosage 2mg/kg, after last gives 30min, be applied to auris dextra auricle both sides with 25 μ l dimethylbenzene, dislocation is put to death behind the 15min, with the card punch of 6mm the punching of mice ears same area is drawn materials, and precision is weighed respectively, with two ear weight differences as swelling degree index, the difference of comparative drug group and group of solvents, and obtain suppression ratio, the results are shown in Table 8.
The influence of table 8 xylol induced mice auricle edema (n=10, x ± s)
| Group | Auricle heavy (mg) | Suppression ratio (%) | |
| A left side | Right | ||
| Blank group aescin for injection nanometer aescine aqueous injection | 10.42±0.67 10.43±0.50 10.53±0.64 | 25.12±0.28 17.18±0.46 * 12.45±0.34 ** | - 31.61 50.44 |
| Nanometer aescine infusion solution nanometer aescine lyophilized injectable powder | 10.57±0.73 11.40±0.93 | 12.05±2.44 ** 12.51±3.32 ** | 52.03 50.19 |
Compare with the blank group:
*Compare with positive controls p＜0.05
*P＜0.01;
The result shows that nanometer aescine ejection preparation of the present invention has the obvious suppression effect to mice auricle swelling, illustrates that said preparation has tangible antiinflammatory action.
Experiment 2
Influence to rat paw edema due to the Ovum Gallus domesticus album
Adopt rat paw edema to test the resist inflammation on repercussive function of observing nanometer aescine ejection preparation of the present invention.50 of SD rats, body weight 180-220g, male, be divided into five groups at random, i.e. blank group, aescin for injection positive controls, nanometer aescine ejection preparation group of the present invention.Each group is applied to the sufficient sole of the foot by corresponding dosage, be coated with 3 every day before the Yu Zhiyan, continuous five days, each dosage 1.75mg/kg, after the last administration 30 minutes, freshly prepared 1% Ovum Gallus domesticus album 0.05ml caused inflammation at rat paw portion subcutaneous injection, with the measurement of sufficient sole of the foot volume determination instrument cause scorching before and after volume-variation below the rat foot sole of the foot joint, calculate and relatively cause scorching back 0.5h, 1h, 2h, 4h respectively organize rat cause scorching before and after the difference of sufficient volume be the swelling degree, respectively organize the difference of swelling degree, the results are shown in Table 9.
Table 9 pair agar causes influence (n=10, the x ± s) of rat paw edema
| Group | Cause scorching front foot sole of the foot volume (ml) | Cause scorching back different time swelling degree of the paw (ml) | |||
| 0.5h | 1h | 2h | 4h | ||
| Blank group aescin for injection nanometer aescine aqueous injection nanometer aescine infusion solution nanometer aescine lyophilized injectable powder | 1.14±0.12 1.13±0.11 1.12±0.13 1.14±0.10 1.15±0.10 | 0.287±0.045 0.203±0.047 * 0.153±0.043 ** 0.162±0.045 ** 0.158±0.048 ** | 0.254±0.035 0.183±0.038 * 0.124±0.037 ** 0.131±0.036 ** 0.129±0.032 ** | 0.215±0.045 0.152±0.046 * 0.086±0.043 ** 0.092±0.048 ** 0.098±0.042 ** | 0.189±0.036 0.112±0.047 * 0.045±0.032 ** 0.053±0.042 ** 0.051±0.041 ** |
Compare with the blank group:
*Compare with positive controls p＜0.05
*P＜0.01;
The result shows that nanometer aescine ejection preparation group of the present invention 1h-4h after administration all suppresses the effect of rat paw edema, illustrates that said preparation has tangible detumescence effect.
Experiment 3
Nanometer aescine ejection preparation is to the inhibitory action of the hemorrhage associated with hydrocephalus of rat brain
Experimental technique: get body weight 180-220g, 60 of male rats are divided into blank group, cerebral hemorrhage mold group negative control group at random, aescin for injection positive controls and reagent group, 10 of every treated animals.Except that normal group, behind animal via pentobarbital sodium (35mg/kg) intraperitoneal injection of anesthesia, head is fixed on the stereotactic apparatus, cut top skin, with reference to the rat brain stereotaxic atlas, press the method for Rosenberg, cerebral hemorrhage group and each treatment group inject collagenase 0.4IU/2 μ l under stereotaxis left side tail shell nuclear, and matched group then injects the equal-volume normal saline; When injecting behind the collagenase 4h, positive controls lumbar injection aescine sodium injection 2.25mg/kg, reagent group lumbar injection nanometer aescine ejection preparation 2.25mg/kg; Sacrificed by decapitation animal when 24h is taken out cerebral hemisphere rapidly, and equal length is three sections before, during and after being divided into by left and right brain, adopts and does wet method mensuration brain water content, the results are shown in Table 10.
The variation of different brains district water content before and after the treatment of table 10 nanometer aescine ejection preparation
| Group | Cortex of parietal lobe | The middle period cortex | The posterior lobe cortex | |||
| A left side | Right | A left side | Right | A left side | Right | |
| Normal saline cerebral hemorrhage mold group aescin for injection nanometer aescine aqueous injection nanometer aescine lyophilized injectable powder | 75.3±0.2 83.4±0.1 79.8±0.2 * 75.8±0.3 * 76.1±0.3 * | 75.4±0.1 83.5±0.2 79.4±0.2 * 75.5±0.5 * 76.2±0.5 * | 75.5±0.27 82.3±0.2 78.5±0.3 * 75.8±0.3 * 76.4±0.3 * | 5.5±0.1 82.4±0.1 78.7±0.2 * 75.5±0.5 * 76.5±0.2 * | 75.8±0.3 83.5±0.1 78.9±0.2 * 75.8±0.3 * 75.8±0.3 * | 75.7±0.5 83.4±0.2 78.8±0.3 * 75.5±0.5 * 75.9±0.2 * |
| Nanometer aescine infusion solution | 75.4±0.5 * | 75.5±0.3 * | 75.9±0.3 * | 75.7±0.5 * | 75.8±0.3 * | 75.6±0.5 * |
Annotate: compare with negative control group
*P＜0.05;
The result shows that under same dose, test group and negative control group comparing difference have the significance meaning, and experimental result explanation nanometer aescine ejection preparation is better than aescin for injection to the inhibitory action of the hemorrhage associated with hydrocephalus of rat brain.
Five, preparation embodiment
Embodiment 1
(1) extraction of aescine effective site:
Get Semen Aesculi 503 grams, pulverize, put into the supersound extraction jar, 25 ℃ of temperature are extracted in control, extract each 5 minutes 2 times, residue is standby, merge extractive liquid, concentrates 40 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.1 atmospheric pressure, relative density is 1.15 when being concentrated to 40 ℃, and concentrated solution was left standstill 24 hours at 0 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make that to contain the alcohol amount be 50%, put into the supersound extraction jar, 25 ℃ of control temperature, extract 2 times, each 15 minutes, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 40 ℃ of control thickening temperatures, vacuum is 0.1 atmospheric pressure, obtains concentrated solution; Get AB-8 type macroporous adsorptive resins on the above-mentioned concentrated solution, earlier with the distilled water eluting of 4 times column volume, eluent discards, the ethanol elution of reuse 60%, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 40 ℃ of control thickening temperatures, vacuum is 0.1 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site 8 grams; [β in the aescine effective site-aescine content is 85%.】
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 8 grams, lipid stearic acid 48 grams, emulsifying agent 32 soybean lecithins gram;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 70 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 70 ℃ of logical condition of nitrogen gas, spare 5-7 time in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, 1000 bottles of preparation cost invention nanometer aescine hydro-acupuncture preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, sterilization, 1000 bottles of preparation cost invention nanometer aescine infusion preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and 1000 bottles of preparation cost invention nanometer aescine freeze-dried powders.[nanometer particle size is the 10-120 nanometer]
Embodiment 2
(1) extraction of aescine effective site:
Get Semen Aesculi 632 grams, pulverize, put into the supersound extraction jar, 50 ℃ of temperature are extracted in control, extract each 15 minutes 4 times, residue is standby, merge extractive liquid, concentrates 50 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.2 atmospheric pressure, relative density is 1.25 when being concentrated to 50 ℃, and concentrated solution was left standstill 48 hours at 4 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make that to contain the alcohol amount be 70%, put into the supersound extraction jar, 50 ℃ of control temperature, extract 4 times, each 45 minutes, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 50 ℃ of control thickening temperatures, vacuum is 0.2 atmospheric pressure, obtains concentrated solution; Get AB-8 type macroporous adsorptive resins on the above-mentioned concentrated solution, earlier with the distilled water eluting of 8 times column volume, eluent discards, the ethanol elution of reuse 90%, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 50 ℃ of control thickening temperatures, vacuum is 0.2 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site 12 grams; [β in the aescine effective site-aescine content is 50%.】
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 12 grams, lipid stearic acid 142 grams, emulsifying agent soybean lecithin 58 grams;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 90 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 90 ℃ of logical condition of nitrogen gas, spare 7 times in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, 1000 bottles of preparation cost invention nanometer aescine hydro-acupuncture preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, sterilization, 1000 bottles of preparation cost invention nanometer aescine infusion preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and 1000 bottles of preparation cost invention nanometer aescine freeze-dried powders.[nanometer particle size is the 10-120 nanometer]
Embodiment 3
(1) extraction of aescine effective site:
Get Semen Aesculi 571 grams, pulverize, put into the supersound extraction jar, 40 ℃ of temperature are extracted in control, extract each 10 minutes 3 times, residue is standby, merge extractive liquid, concentrates 45 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.15 atmospheric pressure, relative density is 1.20 when being concentrated to 45 ℃, and concentrated solution was left standstill 36 hours at 2 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make that to contain the alcohol amount be 60%, put into the supersound extraction jar, 40 ℃ of control temperature, extract 3 times, each 30 minutes, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 45 ℃ of control thickening temperatures, vacuum is 0.15 atmospheric pressure, obtains concentrated solution; Get AB-8 type macroporous adsorptive resins on the above-mentioned concentrated solution, earlier with the distilled water eluting of 6 times column volume, eluent discards, the ethanol elution of reuse 75%, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 45 ℃ of control thickening temperatures, vacuum is 0.15 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site 10 grams; [β in the aescine effective site-aescine content is 72.3%.】
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 10 grams, lipid stearic acid 95 grams, emulsifying agent soybean lecithin 46 grams;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 80 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 80 ℃ of logical condition of nitrogen gas, spare 6 times in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, 1000 bottles of preparation cost invention nanometer aescine hydro-acupuncture preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, sterilization, 1000 bottles of preparation cost invention nanometer aescine infusion preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and 1000 bottles of preparation cost invention nanometer aescine freeze-dried powders.[nanometer particle size is the 10-120 nanometer]
Embodiment 4
(1) extraction of aescine effective site:
Get Semen Aesculi 529 grams, pulverize, put into the supersound extraction jar, 30 ℃ of temperature are extracted in control, extract each 8 minutes 2 times, residue is standby, merge extractive liquid, concentrates 42 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.12 atmospheric pressure, relative density is 1.18 when being concentrated to 43 ℃, and concentrated solution was left standstill 30 hours at 1 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make that to contain the alcohol amount be 55%, put into the supersound extraction jar, 30 ℃ of control temperature, extract 2 times, each 20 minutes, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 42 ℃ of control thickening temperatures, vacuum is 0.13 atmospheric pressure, obtains concentrated solution; Get AB-8 type macroporous adsorptive resins on the above-mentioned concentrated solution, earlier with the distilled water eluting of 5 times column volume, eluent discards, the ethanol elution of reuse 65%, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 43 ℃ of control thickening temperatures, vacuum is 0.13 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site 9 grams; [β in the aescine effective site-aescine content is 80%.】
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 9 grams, lipid stearic acid 56 grams, emulsifying agent soybean lecithin 40 grams;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 75 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 75 ℃ of logical condition of nitrogen gas, spare 5 times in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, 1000 bottles of preparation cost invention nanometer aescine hydro-acupuncture preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, sterilization, 1000 bottles of preparation cost invention nanometer aescine infusion preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and 1000 bottles of preparation cost invention nanometer aescine freeze-dried powders.[nanometer particle size is the 10-120 nanometer]
Embodiment 5
(1) extraction of aescine effective site:
Get Semen Aesculi 611 grams, pulverize, put into the supersound extraction jar, 45 ℃ of temperature are extracted in control, extract each 14 minutes 4 times, residue is standby, merge extractive liquid, concentrates 48 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.19 atmospheric pressure, relative density is 1.24 when being concentrated to 48 ℃, and concentrated solution was left standstill 40 hours at 3 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make that to contain the alcohol amount be 65%, put into the supersound extraction jar, 45 ℃ of control temperature, extract 3 times, each 40 minutes, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 48 ℃ of control thickening temperatures, vacuum is 0.19 atmospheric pressure, obtains concentrated solution; Get AB-8 type macroporous adsorptive resins on the above-mentioned concentrated solution, earlier with the distilled water eluting of 7 times column volume, eluent discards, the ethanol elution of reuse 85%, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 49 ℃ of control thickening temperatures, vacuum is 0.18 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site 11 grams; [β in the aescine effective site-aescine content is 55%.】
(2) preparation of nanometer aescine:
Prescription is: aescine effective site 11 grams, lipid stearic acid 130 grams, emulsifying agent soybean lecithin 48 grams;
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 85 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 85 ℃ of logical condition of nitrogen gas, spare 5-7 time in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(3) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, 1000 bottles of preparation cost invention nanometer aescine hydro-acupuncture preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, sterilization, 1000 bottles of preparation cost invention nanometer aescine infusion preparations; [nanometer particle size is the 10-120 nanometer]
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and 1000 bottles of preparation cost invention nanometer aescine freeze-dried powders.[nanometer particle size is the 10-120 nanometer].
Claims (4)
1. nanometer aescine ejection preparation, it is characterized in that it is by aescine effective site 8-12 weight portion, lipid 48-142 weight portion, emulsifying agent 32-58 weight portion are prepared into, and wherein β in the aescine effective site-aescine content is 50%-85%; Its feature is that also nanometer particle size is the 10-120 nanometer in the nanometer aescine ejection preparation.
2. a kind of nanometer aescine ejection preparation according to claim 1, its preparation method may further comprise the steps:
(1) extraction of aescine effective site:
Get Semen Aesculi, pulverize, put into the supersound extraction jar, 25 ℃-50 ℃ of temperature are extracted in control, extract each 5-15 minute 2-4 time, residue is standby, merge extractive liquid, concentrates 40 ℃-50 ℃ of control thickening temperatures in the rotation knifing instrument, vacuum is 0.1-0.2 atmospheric pressure, relative density is 1.15-1.25 when being concentrated to 40 ℃-50 ℃, and concentrated solution was left standstill 24-48 hour at 0 ℃-4 ℃, removes the oil layer on concentrated solution surface, concentrated solution and the residue of removing oil layer are merged, add dehydrated alcohol, make to contain the alcohol amount, put into the supersound extraction jar for 50%-70%, 25 ℃-50 ℃ of control temperature, extract 2-4 time, each 15-45 minute, merge extractive liquid,, concentrate ethanol to most in the rotation knifing instrument, 40 ℃-50 ℃ of control thickening temperatures, vacuum is 0.1-0.2 atmospheric pressure, obtains concentrated solution; Get macroporous adsorptive resins on the above-mentioned concentrated solution, with the distilled water eluting of 4-8 column volume doubly, eluent discards, the ethanol elution of reuse 60%-90% earlier, the control eluent does not have the saponin reaction, collect eluent, concentrate ethanol to most in the rotation knifing instrument, 40 ℃-50 ℃ of control thickening temperatures, vacuum is 0.1-0.2 atmospheric pressure, obtain concentrated solution, cold drying obtains aescine effective site;
(2) preparation of nanometer aescine:
The glycerol and the poloxamer-188 that get equal in quality are prepared into saturated aqueous solution; Get recipe quantity aescine, emulsifying agent and lipid, under logical condition of nitrogen gas, be heated to 70-90 ℃, at following saturated aqueous solution that adds uniform temp glycerol and poloxamer-188 of stirring condition, make thick breast, under 70-90 ℃ of logical condition of nitrogen gas, spare 5-7 time in 41.4MPa pressure stimulating milk secretion with the high pressure dispersing emulsification machine, after the inflated with nitrogen packing, cooling forms the principal agent suspension rapidly;
(4) formulation preparation:
The preparation of hydro-acupuncture preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, preparation cost invention nanometer aescine hydro-acupuncture preparation;
The preparation of infusion preparation: get above-mentioned suspension, add PVP, add to the full amount of water for injection, stir evenly, add sodium chloride or glucose accent etc. and ooze, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, and sterilization, preparation cost invention nanometer aescine infusion preparation;
The preparation of freeze-dried powder: get above-mentioned suspension, add excipient, add the injection water and adjust concentration, adjust pH is 6.5-7.5, with 0.22 μ m filtering with microporous membrane, lyophilization, and preparation cost invention nanometer aescine freeze-dried powder.
3. be stearic acid according to the lipid in the preparation method of claim 1 and 2 one of described a kind of nanometer aescine ejection preparations.
4. be soybean lecithin according to emulsifying agent in the preparation method of claim 1 and 2 one of described a kind of nanometer aescine ejection preparations.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510089090 CN1907291A (en) | 2005-08-05 | 2005-08-05 | Injectio for nanometer notoginsenoside and its preparing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510089090 CN1907291A (en) | 2005-08-05 | 2005-08-05 | Injectio for nanometer notoginsenoside and its preparing method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1907291A true CN1907291A (en) | 2007-02-07 |
Family
ID=37698628
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510089090 Pending CN1907291A (en) | 2005-08-05 | 2005-08-05 | Injectio for nanometer notoginsenoside and its preparing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1907291A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526575A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel five-flavor decoction integrated dosage form |
| CN102526513A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated Xingshuang decoction dosage form |
| CN102526648A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology for new Xiaolizhong decoction integration type formulation, and production method thereof |
| CN102526650A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated poppy decoction dosage form |
| CN102526647A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel dual-purpose decoction integrated dosage form |
| CN102526649A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Actinolite pill integrative new formulation preparation technology and production method thereof |
| CN102526651A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated Yumi decoction dosage form |
| CN102526646A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel sweet osmanthus decoction integrated dosage form |
| CN102526604A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated costustoot decoction dosage form |
| CN111249226A (en) * | 2020-03-24 | 2020-06-09 | 中南大学 | Aescin injectable hydrogel and preparation method and application thereof |
-
2005
- 2005-08-05 CN CN 200510089090 patent/CN1907291A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526575A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel five-flavor decoction integrated dosage form |
| CN102526513A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated Xingshuang decoction dosage form |
| CN102526648A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology for new Xiaolizhong decoction integration type formulation, and production method thereof |
| CN102526650A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated poppy decoction dosage form |
| CN102526647A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel dual-purpose decoction integrated dosage form |
| CN102526649A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Actinolite pill integrative new formulation preparation technology and production method thereof |
| CN102526651A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated Yumi decoction dosage form |
| CN102526646A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel sweet osmanthus decoction integrated dosage form |
| CN102526604A (en) * | 2010-12-15 | 2012-07-04 | 苏州知微堂生物科技有限公司 | Preparation technology and production method of novel integrated costustoot decoction dosage form |
| CN111249226A (en) * | 2020-03-24 | 2020-06-09 | 中南大学 | Aescin injectable hydrogel and preparation method and application thereof |
| CN111249226B (en) * | 2020-03-24 | 2021-06-15 | 中南大学 | Aescin injectable hydrogel and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1907291A (en) | Injectio for nanometer notoginsenoside and its preparing method | |
| CN1706371A (en) | Efficient sword-like iris seed prepn and its prepn process | |
| CN1939315A (en) | Ganglioside solid lipid nano-particle of monosialic acid tetrahexose and its preparation | |
| CN1947736A (en) | Prepn. method and application of injection contg. Erigeron breviscapus | |
| CN1947713A (en) | Nano-safflower flavone injection prepn. and its prepn. method | |
| KR20170014005A (en) | Coix seed oil comprising 13 glycerides, formulation and application thereof | |
| CN1857385A (en) | Medicine composition for treating cervical spondylosis and its preparing method | |
| CN1923223A (en) | Preparing method and application of nano shen-qi-fu-zheng injectio | |
| CN1891287A (en) | Nano compound musk injection preparion, and its preparing method | |
| CN1903265A (en) | Traditional Chinese medicine composition for treating nycturia and its preparing method | |
| CN1698717A (en) | Chinese medicinal compound fat emulsion injection and its preparation method | |
| CN1775235A (en) | Pseudo-ginseng blood-circulation-invigovating preparation and new preparing method | |
| CN1296069C (en) | Preparing technics of composition of Guizhi and Fuling and its use | |
| CN1879840A (en) | A nano 'Xing Nao Jing' injection and method for preparing same | |
| CN1879639A (en) | A nanometer 'Xue Shuan Tong' injection and preparation method thereof | |
| WO2018120920A1 (en) | Cisatracurium besilate lyophilized powder-injection preparation | |
| CN1899328A (en) | Aescine oral disintegrant tablet and its preparing method | |
| CN1720905A (en) | The application of beta-Asarone in pharmacy | |
| CN102138900B (en) | Solid preparation of oxapium iodide liposome | |
| CN1883545A (en) | A compound nano injection preparation of flavescent sophora root and preparation method thereof | |
| CN1309379C (en) | Asari dripping pills and its preparation process | |
| CN1883471A (en) | A nano sodium cantharidin injection and preparation method thereof | |
| CN1257713C (en) | Flunarizine hydrochloride injection and its preparing method | |
| CN1879638A (en) | A nanometer 'Xue Sai Tong' injection and preparation method thereof | |
| CN1883556A (en) | A nano preparation of 'Kang'ai' injection and method for preparing same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |