CN102366408A - Monosialotetrahexosyl ganglioside sodium liposome injection - Google Patents
Monosialotetrahexosyl ganglioside sodium liposome injection Download PDFInfo
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- CN102366408A CN102366408A CN2011102713283A CN201110271328A CN102366408A CN 102366408 A CN102366408 A CN 102366408A CN 2011102713283 A CN2011102713283 A CN 2011102713283A CN 201110271328 A CN201110271328 A CN 201110271328A CN 102366408 A CN102366408 A CN 102366408A
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Abstract
The invention discloses a monosialotetrahexosyl ganglioside sodium liposome injection and its preparation method. The liposome injection is made from monosialotetrahexosyl ganglioside sodium, hydrogenated egg yolk lecithin, soybean derived sterol, sodium deoxycholate, poloxamer 188, sodium bisulfite and other auxiliary materials. The liposome injection provided by the invention has good stability; the liposome will not break due to fusion and ice crystals during the freezing process and will also maintain good entrapment rate after stored for a long time. According to the invention, the quality of the preparation product is improved, toxic and side effects are minimized, retention time of the medicament in systematic circulation is increased, bioavailability and blood-brain barrier permeability of the medicament are raised and the curative effect is obviously enhanced. In addition, the preparation method is simple and is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of Monostalotetrahexosylgangliside sodium lipidosome and injection and method for making, belong to medical technical field.
Background technology
Monostalotetrahexosylgangliside sodium system in Medulla sus domestica, extract make the neurocyte function damage is had the material of effect, the functional rehabilitation of the central nervous system injury that can promote to cause owing to a variety of causes.The mechanism of action is to promote " neural reconstruct neuroplasticity " (comprising existence, axon growth and the synapse growth of neurocyte).Monostalotetrahexosylgangliside has protective effect to damaging back Secondary cases nerve degeneration.Monostalotetrahexosylgangliside is to cerebral hemodynamic parameter and because of causing cerebral edema that positive effect is arranged after the damage.Monostalotetrahexosylgangliside alleviates the neurocyte edema through the activity of improving the cell membrane enzyme.Zoopery demonstration Monostalotetrahexosylgangliside can improve the behavior disorder due to the parkinson disease.
Monostalotetrahexosylgangliside can also be through keeping the Na on the axoneure film
+-K
+-TTP enzyme and Ca
2+-Mg
2+The activity of-ATP enzyme plays and keeps the inside and outside ionic equilibrium of cell, alleviates the neurocyte edema, prevents the effect that Ca2+ gathers in the cell, has the neurotoxic effect and the effects such as infringement that reduce the radical pair neurocyte of antagonism excitatory amino acid.Exogenous Monostalotetrahexosylgangliside can combine with neuron membrane with stable manner, causes the changes of function of film.Recorded the radioactivity peak at brain and spinal cord in 2 hours after the administration.Reduce by half after 4-8 hour.The removing of medicine is slow, mainly passes through RE.Be mainly used in treatment vascular or traumatic central nervous system injury clinically; Parkinson disease.
Ganglioside molecule water-wet side is close with hydrophobic side length, very easily in water, form stable micelle state, is unfavorable for the preparation of ejection preparation.Patent WO9317691 processes sodium salt with Monostalotetrahexosylgangliside; Injection formulation is cooked buffer with phosphate; Sodium chloride is done isoosmotic adjusting agent, has effectively solved Monostalotetrahexosylgangliside and in water, has formed micellelike state, the difficult problem of medicine preparation.Yet the sodium salt that should write out a prescription influences Monostalotetrahexosylgangliside biomembrane and blood-brain barrier permeability, thereby brain blood drug level is low, and can't maintain for a long time more than the treatment concentration, greatly reduces clinical use curative effect.Therefore be badly in need of seeking a kind of new, can improve Monostalotetrahexosylgangliside sodium and cross biomembrane and blood brain barrier rate, the high method of clinical use curative effect.
A kind of method for preparing and corresponding preparation of Monostalotetrahexosylgangliside are disclosed among the Chinese patent CN101899074A; This patent has mainly been protected the method for preparing of Monostalotetrahexosylgangliside; And corresponding preparation has just been carried out simple explanation, and the preparation of corresponding preparations do not adopt special method yet, causes stability of formulation relatively poor; Because Monostalotetrahexosylgangliside is originally as extract; Stability is very poor, if do not adopt special method, the quality of preparation is certain to very poor.
The compositions of a kind of Monostalotetrahexosylgangliside sodium and glutamic acid is disclosed among the Chinese patent CN101732331A; Replace phosphate buffer and sodium chloride among the patent WO9317691 with glutamic acid; Help Monostalotetrahexosylgangliside sodium through biomembrane and blood brain barrier rate, improve clinical efficacy.But glutamic acid can not play a protective role to the bio-extract Monostalotetrahexosylgangliside equally, and quality of the pharmaceutical preparations stability can not get ensureing equally.
A kind of method for preparing of Monostalotetrahexosylgangliside preparation of sodium is disclosed among the Chinese patent CN1799552A; This patent begins from the original extraction step; Mainly be that whole method for distilling is protected; Corresponding preparation has just been carried out simple description, and preparation do not add any material, and same stability can not get ensureing.
In the pharmaceutical carrier induction system, the research of submicrons such as microemulsion, microsphere, nanoparticle, liposome, pharmacosomes has become field very active in the novel pharmaceutical formulation research.Drug encapsulation can be changed medicine distribution in vivo in these submicrons, increase the abundance of medicine, thereby improve curative effect, alleviate toxic and side effects at target organ.
In targeting drug delivery system, the research of liposome is comparatively extensive, and liposome has good targeting property and biocompatibility in vivo.
As a kind of new medicinal preparation, Liposomal formulation has the following advantages:
(1) have slow releasing function: active component slowly discharges, and delays renal excretion and metabolism, thereby prolongs action time, and effect improves the quality;
(2) reduce drug toxicity;
(3) dissolubility of increase medicine improves the quality of the pharmaceutical preparations;
(4) have targeting property: the contained medicine of liposome is kept high concentration in liver, spleen reticuloendothelial system internal organs part, thereby plays the effect of medicine organ targeting property;
(5) has protective effect to active pharmaceutical ingredient.
Liposome (Liposome) is dispersed in phospholipid by Britain scholar Bangham and Standlish at first and finds when carrying out electron microscopic observation in the water.Phospholipid is dispersed in the water and forms multilamellar vesicle naturally, every layer of equal bilayer of lipid not; Separated by water between vesicle central authorities and each layer, bilayer thickness is about 4nm.Afterwards, this bimolecular folliculus with similar biofilm structure was called liposome.Liposome can be divided into multilamelar liposome and courage and insight liposome.Unilamelar liposome is divided into small unilamellar vesicle and large unilamellar vesicle again.Small unilamellar vesicle is spherical, and size is generally the 20-50 nanometer; Large unilamellar vesicle is of a size of the micron number magnitude.
People such as Britain Lai Men began liposome is used for pharmaceutical carrier in 1971; The main mechanism of action is that drug powder or solution are wrapped in the aqueous phase that the liposome bilayer lipid membrane sealed or embed in the liposome bilayer lipid membrane; This microgranule type of having cellularity; Get into that principal agent is activated the autoimmune function of body in the human body by reticuloendothelial system phagocytic, and change and distributed in the body of entrapped drug, drug main will be put aside in histoorgans such as liver, spleen, lung and bone marrow; Thereby improve the therapeutic index of medicine, reduce the therapeutic dose and the toxicity that reduces medicine of medicine.
In recent years; Along with the continuous progress of biotechnology, liposome preparation technology is progressively perfect, and the liposome mechanism of action is further illustrated; Liposome is fit to vivo degradation, avirulence and non-immunogenicity in addition; Particularly great number tested data proof liposome can improve the Drug therapy index, reduces drug toxicity and reduce drug side effect as pharmaceutical carrier, and reduces drug dose, improves advantages such as stability of drug.
In sum; The physico-chemical property that exists in view of Monostalotetrahexosylgangliside sodium in the prior art and aspect bioavailability and blood-brain barrier permeability in the problem that exists; The inventor is through research chronically; After paying creative work, obtained to be superior to the Monostalotetrahexosylgangliside sodium lipidosome injection of prior art.
Summary of the invention
The objective of the invention is problem to be solved by this invention is the defective that overcomes prior art; A kind of Monostalotetrahexosylgangliside sodium lipidosome injection and method for making thereof are provided; Purpose is to solve existing Monostalotetrahexosylgangliside preparation of sodium stability problem; Liposome plays a protective role to active component, improves its bioavailability, has further promoted the blood-brain barrier permeability of Monostalotetrahexosylgangliside sodium effectively.
In order to form colory Monostalotetrahexosylgangliside sodium lipidosome injection; Can good compatible with Monostalotetrahexosylgangliside sodium it well be sealed and non-leakage filmogen thereby importantly seek, and seek the respective components that can make liposome form the stable injectable agent.
To achieve these goals; Big quantity research and experiment that the inventor carries out find that Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin, soyasterol, NaTDC, the poloxamer 188 of specified weight proportioning can be processed Monostalotetrahexosylgangliside sodium lipidosome injection, wherein; Envelop rate as the Monostalotetrahexosylgangliside sodium of active constituents of medicine is high; The liposome particle diameter is little and be evenly distributed, and compares the retention time significant prolongation of the active constituents of medicine of preparation of the present invention in the body circulation with Monostalotetrahexosylgangliside sodium injection of the prior art; Good stability; The biocompatibility of medicine is high, and bioavailability obviously improves, and curative effect obviously improves.
One of the object of the invention is to provide a kind of Monostalotetrahexosylgangliside sodium lipidosome; It is mainly processed based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium; Hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part; 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part.
In preferred embodiments, based on the weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 4-10 part; Soyasterol 1.2-5 part; NaTDC 0.5-3 part, 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part.
As the phospholipid that is used to form liposome, can use natural phospholipid and synthetic phospholipid.In the present invention, as the Monostalotetrahexosylgangliside sodium of active constituents of medicine, its good water solubility.To the characteristics of Monostalotetrahexosylgangliside sodium, the inventor is particularly suitable for as basic phospholipid filmogen through discovering hydrogenated yolk lecithin.When using other phospholipid, be difficult to form colory liposome, character such as the envelop rate of liposome, stability and percolation ratio are poor.
In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, for the Monostalotetrahexosylgangliside sodium of 1 weight portion, the consumption of hydrogenated yolk lecithin is the 3-14 weight portion.If the consumption of hydrogenated yolk lecithin is lower than 3 weight portions, then can't form stabilized liposomes; Otherwise if the consumption of the consumption of hydrogenated yolk lecithin is higher than 14 weight portions, then the envelop rate as the Monostalotetrahexosylgangliside sodium of active constituents of medicine descends, and the quality of injection and curative effect reduce.
In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, soyasterol, NaTDC and poloxamer 188 are used to regulate the membrane stability of liposome.
(soybean sterol SS) is the hydrolyzate that the soyasterol glucoside removes glucose residue to soyasterol, and said soyasterol glucoside is the mixture from the sterol glucoside of through the Semen sojae atricolor residue that refines Oleum Glycines, separating.Soyasterol can be regulated the stability of hydrogenated yolk lecithin film, and its regulating action effect to stability is superior to cholesterol.
The inventor is through discovering that in Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, for the Monostalotetrahexosylgangliside sodium of 1 weight portion, the consumption of soyasterol is the 0.8-7 weight portion.If the consumption of soyasterol is lower than 0.8 weight portion, the liposome stability that then forms is low, is easy to seepage; Otherwise if the consumption of the consumption of soyasterol is higher than 7 weight portions, the membrane fluidity of Monostalotetrahexosylgangliside sodium is too high, is wrapped in the intravital Monostalotetrahexosylgangliside sodium of lipid and is easy to discharge, and envelop rate descends.
On the other hand; The inventor discovers; In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, for the Monostalotetrahexosylgangliside sodium of 1 weight portion, the consumption of hydrogenated yolk lecithin is the 4-10 weight portion; The consumption of soyasterol is the 1.2-5 weight portion; The NaTDC consumption is the 0.5-3 weight portion, and the consumption of poloxamer 188 is the 0.8-5 weight portion, and the envelop rate of formed Monostalotetrahexosylgangliside sodium lipidosome injection is high.
In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, use poloxamer 188 further to improve the stability of liposome membrane.Poloxamer 188 is a kind of novel non-ionic surface active agents; When being used for the hydrogenated yolk lecithin duplicature; Can improve the chemical energy between this duplicature; Thereby improve the chemical stability of liposome in waterborne liquid, and then improve the stability of Monostalotetrahexosylgangliside sodium lipidosome injection.
In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, for the Monostalotetrahexosylgangliside sodium of 1 weight portion, the consumption of poloxamer 188 is the 0.5-10 weight portion.If the consumption of poloxamer 188 is lower than 0.5 weight portion; Then cause the stability improvement of Monostalotetrahexosylgangliside sodium lipidosome injection not enough owing to its consumption is low excessively; Otherwise; If the consumption of poloxamer 188 is higher than 10 weight portions, it is too high and cause liposome membrane to be easy to reveal then to be used for its consumption.
Discover; When the Monostalotetrahexosylgangliside sodium that uses above-mentioned specified quantitative, hydrogenated yolk lecithin, NaTDC, soyasterol and poloxamer 188; Can obtain colory Monostalotetrahexosylgangliside sodium lipidosome; Its envelop rate and stability are all very high, and toxicity is low, and bioavailability is high.
One of the object of the invention is to provide a kind of Monostalotetrahexosylgangliside sodium lipidosome injection; Described injection comprises injection and freeze-dried powder, and wherein the injection specification is 2ml:20mg, 2ml:40mg and 5ml:100mg; The specification of freeze-dried powder is 40mg, 100mg.
One of the object of the invention is to provide a kind of Monostalotetrahexosylgangliside sodium lipidosome injection, and it is mainly processed based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part, 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part, sodium chloride 0.4-0.8 part and water for injection are an amount of.
In preferred embodiments, based on the weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 4-10 part, soyasterol 1.2-5 part, NaTDC 0.5-3 part, 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part, sodium chloride 0.45-0.9 part and water for injection are an amount of.
One of the object of the invention is to provide a kind of Monostalotetrahexosylgangliside sodium lipidosome freeze-dried injectable powder, and it is mainly processed based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part, 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part, mannitol 3-12 part.
In preferred embodiments, based on the weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 4-10 part, soyasterol 1.2-5 part, NaTDC 0.5-3 part, 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part, mannitol 5-9 part.
In Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention, use sodium sulfite as antioxidant, be used to form stable injection.Sodium chloride is as one of the most frequently used osmotic pressure regulator, and mannitol is as one of the most frequently used lyophilizing skeleton agent, also can not produce any harmful effect to the stability of liposome.
As one of preferred, above-mentioned described Monostalotetrahexosylgangliside sodium lipidosome injection, each composition weight umber ratio of the lipidosome injection of its preparation UD is:
As one of preferred, above-mentioned described Monostalotetrahexosylgangliside sodium lipidosome injection, each composition weight umber ratio of the lipidosome injection of its preparation UD is:
As one of preferred, above-mentioned described Monostalotetrahexosylgangliside sodium lipidosome injection, each composition weight umber ratio of the lipidosome injection of its preparation UD is:
As one of preferred, the sodium lipidosome freeze-dried injectable powder of above-mentioned described Monostalotetrahexosylgangliside, each composition weight umber ratio of the lipidosome injection of its preparation UD is:
As one of preferred, the sodium lipidosome freeze-dried injectable powder of above-mentioned described Monostalotetrahexosylgangliside, each composition weight umber ratio of the lipidosome injection of its preparation UD is:
One of the object of the invention also is to provide a kind of method for preparing Monostalotetrahexosylgangliside sodium lipidosome injection, and it comprises the steps:
(1) hydrogenated yolk lecithin, soyasterol, poloxamer 188 and NaTDC being dissolved in an amount of volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55-65 ℃ of water bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) Monostalotetrahexosylgangliside sodium, sodium sulfite and sodium chloride are dissolved in the water for injection, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m, filling and sealing, sterilization gets Monostalotetrahexosylgangliside sodium lipidosome injection.
One of the object of the invention also is to provide a kind of method for preparing the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside, and it comprises the steps:
(1) hydrogenated yolk lecithin, soyasterol, poloxamer 188 and NaTDC being dissolved in an amount of volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55-65 ℃ of water bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) Monostalotetrahexosylgangliside sodium, sodium sulfite and mannitol are dissolved in the water for injection, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m; Filtrating was placed under-45 ℃ of conditions freezing 4 hours; Be warming up to-13 ℃ with 1.3 ℃/hour speed again, be incubated 3 hours, be warming up to 30 ℃ again; Be incubated 3 hours to dry, obtain the Monostalotetrahexosylgangliside sodium lipidosome freeze-dried preparation.
Beneficial effect
Lipidosome injection of the present invention has improved the formulation products quality, has reduced toxic and side effects, has good preparation stability, has improved bioavailability of medicament, and curative effect obviously improves; And method for preparing is simple, is suitable for industrialized great production.Liposome can be because of dehydration, fusion, ice crystal etc. break in the refrigerating process, and after the long term storage, liposome keeps good envelop rate equally.
Description of drawings
Below, specify embodiment of the present invention in conjunction with accompanying drawing, wherein:
Fig. 1 representes blood drug level-time relation curve.
Wherein,
Curve 1 expression embodiment 1 time front of blood concentration.
Curve 2 expression embodiment 2 time front of blood concentration.
Curve 3 expression embodiment 3 time front of blood concentration.
Curve 4 expression embodiment 4 time front of blood concentration.
Curve 5 expression embodiment 5 time front of blood concentration.
Curve 6 expression Comparative Examples 1 time front of blood concentration.
Curve 7 expression Comparative Examples 2 time front of blood concentration.
Curve 8 expression Comparative Examples 3 time front of blood concentration.
The routine time front of blood concentration of curve 9 expression listings.
The specific embodiment
Below through concrete preferred embodiment the present invention is further specified.These embodiment only are illustrative, and should not be construed as limitation of the present invention.
Embodiment 1The preparation of Monostalotetrahexosylgangliside sodium lipidosome injection
Prescription (1000)
Preparation technology:
(1) 140g hydrogenated yolk lecithin, 60g soyasterol, 50g poloxamer 188 and 36g NaTDC being dissolved in the 2000ml volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 65 ℃ of waters bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) 20g Monostalotetrahexosylgangliside sodium, 0.6g sodium sulfite and 18g sodium chloride are dissolved in the 600ml water, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m, filling and sealing, sterilization gets Monostalotetrahexosylgangliside sodium lipidosome injection.
Embodiment 2The preparation of Monostalotetrahexosylgangliside sodium lipidosome injection
Prescription (1000)
Preparation technology:
(1) 400g hydrogenated yolk lecithin, 200g soyasterol, 200g poloxamer 188 and 120g NaTDC being dissolved in the 6000ml volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55 ℃ of waters bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) 40g Monostalotetrahexosylgangliside sodium, 1.6g sodium sulfite and 18g sodium chloride are dissolved in the 600ml water, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m, filling and sealing, sterilization gets Monostalotetrahexosylgangliside sodium lipidosome injection.
Embodiment 3The preparation of Monostalotetrahexosylgangliside sodium lipidosome injection
Prescription (1000)
Preparation technology:
(1) 800g hydrogenated yolk lecithin, 360g soyasterol, 450g poloxamer 188 and 190g NaTDC being dissolved in the 15000ml volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 60 ℃ of waters bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) 100g Monostalotetrahexosylgangliside sodium, 2g sodium sulfite and 45g sodium chloride are dissolved in the 800ml water, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m, filling and sealing, sterilization gets Monostalotetrahexosylgangliside sodium lipidosome injection.
Embodiment 4The preparation of the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside
Prescription (1000 bottles)
Preparation technology:
(1) 160g hydrogenated yolk lecithin, 48g soyasterol, 32g poloxamer 188 and 20g NaTDC being dissolved in the 4000ml volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55 ℃ of waters bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) 40g Monostalotetrahexosylgangliside sodium, 0.8g sodium sulfite and 200g mannitol are dissolved in the 1200ml water, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m; Filtrating was placed under-45 ℃ of conditions freezing 4 hours; Be warming up to-13 ℃ with 1.3 ℃/hour speed again, be incubated 3 hours, be warming up to 30 ℃ again; Be incubated 3 hours to dry, obtain the Monostalotetrahexosylgangliside sodium lipidosome freeze-dried preparation.
Embodiment 5The preparation of the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside
Prescription (1000 bottles)
Preparation technology:
(1) 500g hydrogenated yolk lecithin, 150g soyasterol, 100g poloxamer 188 and 60g NaTDC being dissolved in the 8000ml volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 65 ℃ of waters bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) 100g Monostalotetrahexosylgangliside sodium, 3g sodium sulfite and 900g mannitol are dissolved in the 2000ml water, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m; Filtrating was placed under-45 ℃ of conditions freezing 4 hours; Be warming up to-13 ℃ with 1.3 ℃/hour speed again, be incubated 3 hours, be warming up to 30 ℃ again; Be incubated 3 hours to dry, obtain the Monostalotetrahexosylgangliside sodium lipidosome freeze-dried preparation.
Comparative Examples 1-3The preparation of Monostalotetrahexosylgangliside sodium lipidosome injection
Adopt the production technology identical with embodiment 1; Component among the Comparative Examples 1-2 as shown in the table and consumption are processed Monostalotetrahexosylgangliside sodium lipidosome injection respectively; Adopt the production technology of similar embodiment 4, component in the following table Comparative Examples 3 and consumption are processed the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside respectively.
Table 1 Comparative Examples 1-3 component
Wherein, "/" expression is not used.
Test Example 1The mensuration of liposome particle diameter
Under the room temperature condition; Get the Monostalotetrahexosylgangliside sodium lipidosome injection among embodiment 1-5 and the Comparative Examples 1-3; Place the sample cell of Submicron Particle Sizer Model 370 particle diameter detectors, measure particle size distribution and mean diameter; Observe particle shape with projection electron microscope.The result is shown in the following table 2.
The mensuration of table 2 particle diameter
Can know that from table 2 the liposome particle diameter that embodiment 1-5 makes is even, show spherical, big or small homogeneous; The liposome particle diameter that Comparative Examples 1-3 makes is inhomogeneous, and shape is indefinite, and is disorderly and unsystematic, not of uniform size.
Particularly; Even when adopting same production technology, the particle appearance of the Monostalotetrahexosylgangliside sodium lipidosome that makes among the embodiment 1-5 and mean diameter thereof obviously are superior to the Monostalotetrahexosylgangliside sodium lipidosome that makes among the Comparative Examples 1-3.When the composition beyond using the used composition of the present invention was described, perhaps when the composition consumption was outside the composition amount ranges that the present invention limits, the outward appearance of gained Monostalotetrahexosylgangliside sodium lipidosome was inferior to the present invention, and mean diameter obviously goes out greatly a lot.
Test Example 2The mensuration of envelop rate
With the rotating speed high speed centrifugation of the Monostalotetrahexosylgangliside sodium lipidosome injection for preparing among embodiment 1-5 and the Comparative Examples 1-3 with 5000r/min; Centrifugal 20 minutes; Get supernatant, use dissolve with methanol, the HPLC method is surveyed the Monostalotetrahexosylgangliside sodium content; Computational envelope rate, result are shown in the following table 3.
Table 3 entrapment efficiency determination result
Can know that by table 3 envelop rate of the Liposomal formulation of embodiment 1-5 preparation is higher than the envelop rate of the Liposomal formulation of Comparative Examples 1-3 significantly.When the composition beyond using the used composition of the present invention was described, perhaps when the composition consumption was outside the composition amount ranges that the present invention limits, the liposome encapsulation of gained liposome was lower than the present invention.
Test Example 3Study on the stability
The sample that the sample of embodiment of the invention 1-5 preparation and Comparative Examples 1-3 are prepared, Monostalotetrahexosylgangliside sodium injection (the lot number 20100303-1 of listing; The Beijing Sihuan Pharmaceutical Co., Ltd) places following 6 months of the condition of 40 ℃ of high temperature, relative humidity 75% respectively; Carry out accelerated test and investigate, result of the test is seen table 4.
Table 4 accelerated test result
Can be known that by table 4 when quickening June, the sample of Comparative Examples 1-3 preparation reduces with the listing formulation content, related substance raises, and pH value descends obviously; And the sample character of embodiment of the invention 1-5, pH value, content and related substance variation are all not obvious, explain that product stability of the present invention is good.
Test Example 4The percolation ratio test
Get the sample of Test Example 1-5 and Comparative Examples 1-3 preparation, at ambient temperature,, make regular check on, measure envelop rate respectively at 0 day, 30 days, 60 days, 90 days and 180 days, with the dose of sealing in 0 day relatively, calculate percolation ratio, the result is shown in the following table 5.
Table 5 percolation ratio result of the test
Can know by table 5; During long term storage; The Monostalotetrahexosylgangliside sodium lipidosome injection percolation ratio for preparing among the embodiment of the invention 1-5 changes little; And the injection percolation ratio for preparing among the Comparative Examples 1-3 increases gradually, and the liposome seepage is serious, and the Monostalotetrahexosylgangliside sodium lipidosome injection of this explanation the inventive method preparation has higher stability.
Test Example 5The mensuration of blood drug level
90 rats are divided into 9 groups at random; Every group of sample for preparing among drug administration by injection embodiment 1-5 and the Comparative Examples 1-3 respectively; And commercially available Monostalotetrahexosylgangliside sodium injection (lot number 20100303-1; The Beijing Sihuan Pharmaceutical Co., Ltd), injection volume is a 20mg Monostalotetrahexosylgangliside sodium.Respectively at 0h, 0.5h, 1h, 1.5h, 2h, 3h, 6h, 8h, take a blood sample after the administration, blood sample is measured blood drug level with the HPLC-MS method after treatment.The Monostalotetrahexosylgangliside sodium lipidosome injection for preparing among the Monostalotetrahexosylgangliside sodium lipidosome injection for preparing among the drafting embodiment 1-5, the Comparative Examples 1-3 and the blood drug level and the time relation curve of commercially available Monostalotetrahexosylgangliside sodium injection are shown in the accompanying drawing 1.
Wherein,
Curve 1 expression embodiment 1 time front of blood concentration.
Curve 2 expression embodiment 2 time front of blood concentration.
Curve 3 expression embodiment 3 time front of blood concentration.
Curve 4 expression embodiment 4 time front of blood concentration.
Curve 5 expression embodiment 5 time front of blood concentration.
Curve 6 expression Comparative Examples 1 time front of blood concentration.
Curve 7 expression Comparative Examples 2 time front of blood concentration.
Curve 8 expression Comparative Examples 3 time front of blood concentration.
The routine time front of blood concentration of curve 9 expression listings.
Can know by Fig. 1; Compare with commercially available Monostalotetrahexosylgangliside sodium lipidosome injection with the Monostalotetrahexosylgangliside sodium lipidosome injection for preparing among the Comparative Examples 1-3; The Monostalotetrahexosylgangliside sodium lipidosome injection for preparing among the embodiment of the invention 1-5 has the following advantages: elimination speed is in vivo slowed down; Distribution time prolongs in the body circulation, has reached improved slow release effect, and bioavailability increases.
Test Example 6The blood-brain barrier permeability experiment
Get 30 of new zealand rabbits; The about 2.5kg of body weight, male and female are not limit, and are divided into 6 groups at random; The Monostalotetrahexosylgangliside sodium lipidosome injection of 1-5 group intravenous injection embodiments of the invention 1-5; The 6th group of commercially available Monostalotetrahexosylgangliside sodium injection of intravenous injection (lot number 20100303-1, Beijing Sihuan Pharmaceutical Co., Ltd), injection volume are 20mg Monostalotetrahexosylgangliside sodium.Different time extracts the 0.2ml cerebrospinal fluid and makes an experiment after administration, is respectively 0.5h, 1h, 2h, 4h, 8h behind the injectable drug sample time.Get cerebrospinal fluid 0.1ml, add an amount of solvent suspendible 15min,, get supernatant 20ul sample introduction, use high-performance liquid chromatogram determination with the centrifugal 1min of 10000rpm.The result sees table 6.
The mensuration of table 6 cerebrospinal fluid Chinese medicine concentration
Annotate: compare * with the listing example and represent significant difference.
Can know by table 6 result of the test; Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention is significantly improved than the routine injection of listing through the medication amount of blood brain barrier; Explain that Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention obviously increases through the medication amount of blood brain barrier; Improve the blood drug level in the cerebral tissue, improved therapeutic effect.Result of the test also shows; After the Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention administration; The drug level of target tissue raises fast and is stable; In tissue, have long-acting, therefore Monostalotetrahexosylgangliside sodium lipidosome injection of the present invention can reduce administration number of times.And under the identical dosage, the effect of embodiment preparation obviously is superior to go on the market routine preparation and Comparative Examples.
Industrial applicibility
Result by the foregoing description and Test Example can know that Monostalotetrahexosylgangliside sodium lipidosome of the present invention has good surface appearance, and granule is little; Particle diameter is even, and envelop rate is high, and stability is high; Percolation ratio is low; The time of staying in vivo is long, and bioavailability is high, has the favorable industrial using value.
More than through the specific embodiment and Test Example the present invention is specified; But should understand; These explanations do not constitute any restriction to scope of the present invention; Under the situation that does not depart from spirit of the present invention and protection domain, can carry out multiple modification, improvement and replacement to technical scheme of the present invention and embodiment thereof, these are all because of falling in protection scope of the present invention.
Each list of references of mentioning among the application or quoting is incorporated herein by reference at this in full.
Claims (9)
1. Monostalotetrahexosylgangliside sodium lipidosome; It is characterized in that mainly by processing: 1 part in Monostalotetrahexosylgangliside sodium based on the following composition of weight portion meter; Hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part; 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part.
2. Monostalotetrahexosylgangliside sodium lipidosome according to claim 1; It is characterized in that mainly by processing: 1 part in Monostalotetrahexosylgangliside sodium based on the following composition of weight portion meter; Hydrogenated yolk lecithin 4-10 part, soyasterol 1.2-5 part, NaTDC 0.5-3 part; 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part.
3. Monostalotetrahexosylgangliside sodium lipidosome injection, it is characterized in that mainly by processing based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part, 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part, sodium chloride 0.4-0.8 part and water for injection are an amount of.
4. Monostalotetrahexosylgangliside sodium lipidosome injection according to claim 3, it is characterized in that mainly by processing based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 4-10 part, soyasterol 1.2-5 part, NaTDC 0.5-3 part, 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part, sodium chloride 0.45-0.9 part and water for injection are an amount of.
5. the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside is characterized in that mainly by processing based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 3-14 part, soyasterol 0.8-7 part, NaTDC 0.3-5 part, 0.5-10 part poloxamer 188, sodium sulfite 0.01-0.05 part, mannitol 3-12 part.
6. the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside according to claim 5 is characterized in that mainly by processing based on the following composition of weight portion meter: 1 part in Monostalotetrahexosylgangliside sodium, hydrogenated yolk lecithin 4-10 part, soyasterol 1.2-5 part, NaTDC 0.5-3 part, 0.8-5 part poloxamer 188, sodium sulfite 0.02-0.04 part, mannitol 5-9 part.
7. a method for preparing Monostalotetrahexosylgangliside sodium lipidosome injection is characterized in that comprising the steps:
(1) hydrogenated yolk lecithin, soyasterol, poloxamer 188 and NaTDC being dissolved in an amount of volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55-65 ℃ of water bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) Monostalotetrahexosylgangliside sodium, sodium sulfite and sodium chloride are dissolved in the water for injection, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m, filling and sealing, sterilization gets Monostalotetrahexosylgangliside sodium lipidosome injection.
8. a method for preparing the sodium lipidosome freeze-dried injectable powder of Monostalotetrahexosylgangliside is characterized in that comprising the steps:
(1) hydrogenated yolk lecithin, soyasterol, poloxamer 188 and NaTDC being dissolved in an amount of volume ratio is in 1: 3 the isopropyl alcohol and alcohol mixed solvent, a type lipoprotein solution;
(2) above-mentioned type of lipoprotein solution placed the pyriform bottle, in 55-65 ℃ of water bath with thermostatic control, rotary evaporation is removed mixed solvent, forms even lipid membrane;
(3) Monostalotetrahexosylgangliside sodium, sodium sulfite and mannitol are dissolved in the water for injection, add jog in the pyriform bottle, make the lipid membrane eluting and be distributed to the hydration medium dissolving, promptly get liposome turbid liquor;
(4) place Ultrasound Instrument ultrasonic above-mentioned suspension to translucent colloid solution;
(5) with the filtering with microporous membrane of above-mentioned suspension with 0.45 μ m; Filtrating was placed under-45 ℃ of conditions freezing 4 hours; Be warming up to-13 ℃ with 1.3 ℃/hour speed again, be incubated 3 hours, be warming up to 30 ℃ again; Be incubated 3 hours to dry, obtain the Monostalotetrahexosylgangliside sodium lipidosome freeze-dried preparation.
9. the described Monostalotetrahexosylgangliside sodium lipidosome of claim 1 improves the purposes in the medicine of blood-brain barrier permeability of Monostalotetrahexosylgangliside sodium in preparation.
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Cited By (5)
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| CN102988284A (en) * | 2012-12-13 | 2013-03-27 | 哈药集团技术中心 | Preparation method for monosialotetrahexosyl ganglioside sodium injection |
| US20150064115A1 (en) * | 2012-04-05 | 2015-03-05 | University Of Florida Research Foundation, Inc. | Neurophilic nanoparticles |
| CN104473885A (en) * | 2014-12-09 | 2015-04-01 | 山东新时代药业有限公司 | Monosialotetrahexosyl ganglioside sodium sterile powder injection |
| CN110755405A (en) * | 2019-10-25 | 2020-02-07 | 南昌大学 | Application of ganglioside GM3 with targeting property in preparing medicine for treating atherosclerosis |
| CN113304276A (en) * | 2021-06-04 | 2021-08-27 | 沈阳药科大学 | Liposome modified by monosialotetrahexosylganglioside, preparation method and freeze-drying application thereof |
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