CN100370968C - Quercetin long-acting liposome powder for injection and its preparing method - Google Patents
Quercetin long-acting liposome powder for injection and its preparing method Download PDFInfo
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Abstract
The present invention relates to a long acting liposome powder injection of quercetin and a preparation method thereof. The powder injection is composed of quercetin, polyethylene glycol-phosphatidyl ethanolamine, lecithin, cholesteryl and excipient. The preparation method of the powder injection comprises the steps that 5 to 15 weight parts of polyethylene glycol-phosphatidyl ethanolamine, 40 to 60 weight parts of lecithin, 5 to 15 weight parts of cholesteryl and 20 to 50 weight parts of quercetin are dissolved in an organic solvent with the proportion of trichloromethane to carbinol of 1 to 4: 1; a long acting liposome solution of the quercetin is obtained by the existing thin film ultrasonic method; the excipient is added in the solution, and the solution is dried in a freezing mode to obtain the long acting liposome powder injection of quercetin. The long acting liposome powder injection of quercetin, which is obtained by the present invention with optimum technology parameters can thoroughly dissolve in a solvent for intravenous drip infusion; the absorption of the quercetin in a body is improved, the blood circulation time in a body is prolonged, and the bioavailability of the quercetin is improved.
Description
One, technical field
The present invention relates to a kind of quercetin long-acting liposome powder for injection and preparation method thereof, more particularly, relate to a kind of employing Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE (PEG-PE) and modify the surperficial long-acting liposome that gets of conventional liposome, a kind of quercetin long-acting liposome powder for injection of the Quercetin of long-acting liposome parcel biologically active and preparation method thereof.
Two, background technology
Quercetin (3,3,4,5,7-phentahydroxyflavone) and derivant be plant kingdom's the widest flavone compound that distributes, extensively be present in (Frankel et al., 1993 such as edible vegetable and fruit such as Fructus Crataegi, Fructus Mali pumilae, Bulbus Allii Cepae, Folium Camelliae sinensis, Mel, Fructus Vitis viniferae; Hertog et al., 1993,1996) and among multiple Chinese herbal medicine such as Cacumen Platycladi (Cacumen Biotae), Rhizoma Alpiniae Officinarum, Flos Farfarae, Radix Notoginseng, Semen Ginkgo etc. (Formica JV et al., 1995).Quercetin also has the same principle construction the same with other flavone compound, is to be made of three ring molecules that connect hydroxyl, and the chemical structural formula of Quercetin is:
Because the aboundresources of Quercetin is easy to get, and have very strong biological activity and pharmacological action widely and extremely attention.Since the beginning of this century, Chinese scholars has been carried out number of research projects to the extraction of Quercetin, purification, structure determination, physicochemical property, synthesis of derivatives, pharmacology and aspect such as clinical.The multiple biological activity and the pharmacological action of relevant Quercetin are confirmed, and it has antioxidant activity (Hayek T et al., 1997; Chopra M et al., 2000), anticancer (VermaAK et al., 1988; Deschner EE et al., 1991; Pereira MA et al., 1996), anti-inflammatory (Ferry DRet al., 1996), anti-(Pignatelli P et al., 2000), the expansion coronary vasodilator effects such as (Perez-Vizcaino F etal., 2002) of assembling.
But, because the Quercetin that extracts in the plant is insoluble in water and many medicinal solvents, thereby utilize Quercetin to prepare difficulty of ideal pharmaceutical formulation, and then hindered Quercetin in Clinical Application.For improving the water solublity of Quercetin, improve its absorption in vivo, domestic and international research persons have taked several different methods, as Quercetin being dissolved in the dimethyl sulfoxine organic solvent, be used for clinical research, but, and can give off an unpleasant smell after using and be terminated (Muther RS et al., 1980 because the dosage of dimethyl sulfoxine crosses conference and cause the intravital globulolysis of people; Marshall LF et al., 1984).And for example, researcheres also carry out suitable modification to the chemical constitution of Quercetin, employing is introduced hydrophilic group on its structure, the water soluble derivative of preparation Quercetin, operation process such as its absorption in vivo, distribution, metabolism and drainage are improved, bring into play its pharmacological action (P.J.Mulholland et al., 2001).But the method for taking chemical modification makes the pharmacological action of Quercetin be affected, and there are problems such as separation, purification in chemical modification.Therefore, also do not have at present suitable and economic approach with Quercetin as medicinal application in clinical and suitability for industrialized production.This just presses for the preparation formulation that changes Quercetin, selects ideal drug administration carrier to make and can overcome the defective of Quercetin water solublity and pharmacokinetics difference, and make the active drug targeting in liver, intestinal position, can not bring side effect such as anaphylaxis again.The present inventor then is the present invention who makes at this problem.
As everybody knows, liposome belongs to a kind of novel form of targeting drug delivery system as the targeted drug carrier.1173) etc. at the end of the sixties, (1979,146 (4): the people at first uses liposome Rahman as pharmaceutical carrier for Rahman Y E, et al.Proc Soc Exp Biol Med.Because liposome has the class cellularity, can change interior distribution of body of encapsulated medicine, drug main to be accumulated in histoorgans such as liver, spleen, lung and bone marrow, thereby improve curative effect of medication, reduce Drug therapy dosage, reduce drug toxicity.Therefore liposome has shown its obvious superiority as the carrier of multiclass medicine in the numerous disease treatment.
Long-acting liposome normally mixes some degradable biological materials on the conventional liposome surface, as Polyethylene Glycol (PEG) quasi-grease derivative, tween, Brij (Zhang Xiaobin, Hou Xinpul.2003,12 (2): 71-75) etc., the quasi-grease derivative of Polyethylene Glycol (Alexander L.Klibanov wherein, Kazuo Maruyama, et al., 1990,268 (1): 235-237) Zhi Bei long-acting liposome, because of having raw material, it is easy to get, moderate, characteristics such as no immunotoxicity and have superiority, the peg moiety that contains the PEG quasi-grease derivative in the composition of long-acting liposome provides the long-acting liposome water-wetted surface, reduced the interaction of plasma protein and cell surface part, thereby reduce the picked-up of mononuclear phagocyte system (MPS), prolong blood circulation time, make long-acting liposome can effectively arrive diseased region.
Liposome can be by even method (the Hamilton R L J of high pressure breast, Goerke J, et al., 1980,21 (5): 981-992), thin-film ultrasonic method (Huang C, 1969,344), emulsifying dispersion method (Szoke F J, Papahadjopoulos D, etal. 8 (1):, 1978,75 (9): method preparation such as 4194-4198).
Three, summary of the invention
Water soluble derivative existing defective in drug research and application at existing Quercetin and Quercetin thereof, the object of the present invention is to provide a kind of quercetin long-acting liposome powder for injection, this injectable powder not only can make the Quercetin of biologically active can fully be dissolved in the solvent that is used for intravenous drip, thereby prolong Quercetin blood circulation time in vivo, make Quercetin effectively to arrive diseased region, improve its absorption in vivo; Another object of the present invention provides a kind of preparation method of above-mentioned quercetin long-acting liposome powder for injection, and this method is not only improved raw materials used and consumption, optimizes preparation parameter, and easy to operate, is applicable to large-scale continuous production.
The quercetin long-acting liposome powder for injection that basic thought of the present invention provides Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE (PEG-PE), lecithin, cholesterol and the excipient of a kind of Quercetin by biologically active, different molecular quality to form on the one hand; Provide Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of the different molecular quality that a kind of employing has degradable biomaterial to modify the conventional liposome surface that lecithin and cholesterol are formed on the other hand, get long-acting liposome, long-acting liposome parcel Quercetin is as the targeted drug carrier of Quercetin, and improves the method that raw materials used and consumption, preparation parameter prepare quercetin long-acting liposome powder for injection.This method not only obtains the quercetin long-acting liposome of high envelop rate and narrower particle size distribution, and the pharmacological action that has improved Quercetin makes Quercetin easily be dissolved in the medicinal solvent, prolongs its in vivo blood circulation time, improves its absorption in vivo, with research and the application of Quercetin in medicine of development biologically active.
Purpose of the present invention realizes by the technical scheme that is made of following measure.
Quercetin long-acting liposome powder for injection of the present invention, it is by Quercetin 20-50 part, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE 5-15 part, lecithin 40-60 part, cholesterol 5-15 part of biologically active, and excipient 0.1-5 part is formed, wherein the molecular mass of Polyethylene Glycol is 2000-8000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the consumption proportion of lecithin and cholesterol is 3-12: 1, and its component is all by weight.
Used Polyethylene Glycol-the PHOSPHATIDYL ETHANOLAMINE of the present invention is self-control, and synthetic method is according to document: Alexander L.Klibanov, Kazuo Maruyama, et al., 1990,268 (1): the 235-237 gained.
In the such scheme, the molecular weight of Polyethylene Glycol is 2000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 10-15 part.
In the such scheme, the molecular weight of Polyethylene Glycol is 4000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 5-11 part.
In the such scheme, the molecular weight of Polyethylene Glycol is 8000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 8-10 part.
In the such scheme, said excipient is a sorbitol, or mannitol, or glucose, or sucrose, or trehalose.
Realize the preparation method of arbitrary described quercetin long-acting liposome powder for injection in the such scheme, adopt the thin-film ultrasonic method to make, according to the present invention, modify the surperficial long-acting liposome that gets of conventional liposome of lecithin and cholesterol composition with Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of different molecular quality, the Quercetin of long-acting liposome parcel biologically active, this method is raw materials used and consumption, technological parameter and main technique step are as follows:
(1) Quercetin 20-50 part, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE 5-15 part, lecithin 40-60 part, cholesterol 5-15 part, wherein the molecular mass of Polyethylene Glycol is 2000-8000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the proportioning of lecithin and cholesterol is 3-12: 1, above raw material is solid-state, and with milligram or gram or kilogram metering, organic solvent chloroform and methanol are liquid, their volume proportion is 1-4: 1, with milliliter or liter metering;
Quercetin, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, lecithin, the cholesterol that measures is dissolved in the chloroform and methanol organic solvent that proportioning is good according to the above ratio, making the gained solution concentration is 1mg/ml-3mg/ml, place then in the airtight bottle, be stirred to dissolving fully at ambient temperature;
(2) with in the flask that above-mentioned consoluet solution is transferred to rotary evaporator is connected, it is that the evacuation rotary evaporation is removed organic solvent then, forms the layer of even thin film to flask walls in 30 ℃-50 ℃ the water bath with thermostatic control that flask is invaded temperature;
(3) flask that will be formed with thin film is put into vacuum drying oven, dry 2 hours at ambient temperature, further removes residual organic solvent;
(4) aquesterilisa is added in the dried flask, make the complete submergence of thin film of bottle wall, use the Ultrasonic Cell Disruptor ultra-sonic dispersion, its ultrasonic power is 100w-1000w, and ultrasonic time is 0.5-3 hour, promptly gets quercetin long-acting liposome solution;
(5) in quercetin long-acting liposome solution, add excipient, after the lyophilizing, deposit in 4 ℃-8 ℃ cryogenic conditions lower cover sealing and preserve standby.
In the such scheme, the molecular mass of Polyethylene Glycol is 2000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 10-15 part.
In the such scheme, the molecular mass of Polyethylene Glycol is 4000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 5-11 part.
In the such scheme, the molecular mass of Polyethylene Glycol is 8000 o'clock in used Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 8-10 part.
In the such scheme, said excipient is a sorbitol, or mannitol, or glucose, or sucrose, or trehalose.
It is 2000-8000 that the present invention selects the molecular mass of Polyethylene Glycol in the Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of modifying the conventional liposome surface for use, because Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is a straight-chain molecular structure, the increase of chain personal attendant molecular weight and increasing, adding Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE when the preparation long-acting liposome embeds in the liposome membrane it, can effectively prolong Quercetin blood circulation time in vivo, make quercetin long-acting liposome can reach diseased region effectively, improve its absorption in vivo and stable effect, the medicine that has improved Quercetin is for power.
The peg molecule quality is 2000 o'clock in use Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, because molecular mass is less relatively, the molecule segment length of the Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE that forms is shorter, promptly need increase the consumption of Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, it is long-lasting to guarantee in preparation process liposome to be had.Therefore, when to use the peg molecule quality be 2000 Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, addition was 5-15 part, is preferably 10-15 part.
Use that the peg molecule quality is in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE at 4000 o'clock, because its molecular mass is moderate, more help the stability of liposome and long-lasting, therefore, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 5-11 part.
The peg molecule quality is 8000 o'clock in use Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the viscosity of Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE increases along with the increase of molecular weight, if Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE consumption is too big, because its viscosity is big, it is inhomogeneous to make rotary evaporation prepare the thin film of liposome, the size of ultrasonic back liposome particles is uneven, make the poor stability of long-acting liposome, otherwise, reduce and can influence the long-lasting of liposome as the consumption of Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, therefore, when the peg molecule quality is 8000, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part, is preferably 8-10 part.
The mass ratio of used lecithin of the present invention and cholesterol is influential to the liposome size, as shown in Table 1, the weight ratio of lecithin and cholesterol is that the uniformity and the envelop rate of 3: 1 and 4: 1 o'clock liposome particles is better, so the weight ratio of lecithin and cholesterol is preferably 4: 1.
Table 1 lecithin and cholesterol ratio optimization table
| Lecithin/cholesterol (weight ratio, gram or kilogram) | Granular size (nm) |
| 3∶1 | 123±30 |
| 4∶1 | 103±25 |
| 6∶1 | 153±40 |
| 8∶1 | 185±41 |
The phase transition temperature of lecithin is 30 ℃-50 ℃, in lecithin, add the phase transition temperature that cholesterol can change its lecithin, phase transition temperature can influence the shaping of lipid membrane, and then influences the size of liposome particles and evenly, and the water bath with thermostatic control temperature was regulated and control when this temperature can be by rotary evaporation.When preparing quercetin long-acting liposome powder for injection among the present invention, after adding lecithin, cholesterol, Quercetin, the Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the temperature during the preparation lipid membrane is 30 ℃-50 ℃, and preferred temperature is 40 ℃.And cholesterol increases the flowability of film to the flowability generation regulatory function of lipid film, can improve the stability and the envelop rate of liposome.
Used organic solvent chloroform of the present invention and methanol ratio are also influential to the liposome size, and as shown in Table 2, the used optimized proportion of organic solvent chloroform and methanol is 1: 1 and 3: 1, is preferably 3: 1, and volume is with milliliter or rise metering.
Table 2 chloroform (CHCl
3) and methanol (CH
3OH) solvent optimization table
| Chloroform/methanol (volume ratio ml or L) | 1∶1 | 2∶1 | 3∶1 | 4∶1 |
| Granular size (nm) | 136±35 | 162±45 | 137±30 | 148±38 |
The used Ultrasonic Cell Disruptor of the present invention is broken to disperse the time of thin film relevant with its power, prolongs the broken time, increases broken power, prepared liposome particle diameter is more little, but the ultrasonication overlong time, power is excessive, can destroy the integrity of liposome membrane, and then have influence on the envelop rate of medicine.Therefore, the preparation Quercetin selected ultrasonication time of liposome powder for injection is 0.5-3 hour among the present invention, and ultrasonication power is 100-1000W, just can guarantee to obtain particle diameter granularity and high envelop rate preferably.
The employing dextran gel column chromatography detects, select the sephadex column SephadexG-75 dress post of peace agate West Asia for use, the double distilled water eluting with 394nm monitoring and collection eluting peak, knows that by the result who goes out who detects the envelop rate of the quercetin long-acting liposome that the present invention prepares is 95%.
Quercetin long-acting liposome powder for injection of the present invention and preparation method thereof has following characteristics:
1, quercetin long-acting liposome powder for injection of the present invention, Quercetin can fully be dissolved in the medicinal solvent that carries out intravenous drip the glucose solution as 5%, normal saline after wrapping up with long-acting liposome.
2, quercetin long-acting liposome powder for injection of the present invention is dissolved in the medicinal solvent as injection and instils, and not only can improve its absorption in vivo, and can prolong its blood circulation time in vivo, thereby the medicine that further improves Quercetin is for power.
3, but quercetin long-acting liposome powder for injection of the present invention long preservation under 4 ℃～8 ℃ conditions of low temperature is standby, has good stability, sees Table 3.
Table 3 quercetin long-acting liposome powder for injection is in 4～8 ℃ of following stability test data of low temperature
| Lot number | Time (moon) | Outward appearance | Microscopic examination |
| Quercetin long-acting liposome | 0 | The class yellow green shape thing that loosens | The dissolving no crystallization in back is separated out, and does not assemble |
| 1 | The class yellow green shape thing that loosens | The dissolving no crystallization in back is separated out, and does not assemble | |
| 6 | The class yellow green shape thing that loosens | The dissolving no crystallization in back is separated out, and does not assemble | |
| 12 | The class yellow green shape thing that loosens | The dissolving no crystallization in back is separated out, and does not assemble |
4, preparation method provided by the invention, owing to be modified with the conventional liposome surface of lecithin and cholesterol composition with the Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of different molecular quality, and optimization preparation parameter, make long-acting liposome pharmaceutical carrier with higher entrapment and narrower particle size distribution, with the Quercetin of long-acting liposome carrier parcel biologically active, reach the pharmacological action that improves Quercetin, make Quercetin easily be dissolved in the medicinal solvent, prolong its in vivo circulation time, improve its absorption in vivo.
Four, description of drawings
The photo figure of Fig. 1 quercetin long-acting liposome transmission electron microscope of the present invention.
Fig. 2 quercetin long-acting liposome form of the present invention and grading curve distribution schematic diagram.
By the photo of Fig. 1 transmission electron microscope irradiation as can be known, quercetin long-acting liposome film of the present invention is finger print, and helical structure is clear; The particle size distribution range of quercetin long-acting liposome of the present invention is narrower as can be known by the curve distribution of Fig. 2, and mean diameter is 100 ± 25nm.
Five, the specific embodiment
The following examples and biological activity test are to further describe of the present invention, but do not mean that any limitation of the invention.
1, the present invention is to the acute toxicity test of mice:
1. get 20 of BABL/c mices, 6～8 weeks, male and female half and half, fasting 12 hours, normal drinking-water, the mode of employing tail intravenously administrable, the maximal dose of tail intravenously administrable is that the content of Quercetin is 40mg/ml, every dosage is 0.2ml, is equivalent to 480 times of people's consumption per day, and tangible poisoning symptom and death condition do not appear in mice after the medication, observe a week continuously, animal all survives, activity freely, hair smoothing, diet is normal, one week back dissection animal, tangible pathological change does not appear in naked eyes and microscopic examination internal organs.
2, pharmacokinetics experiment of the present invention:
1. get 20 of BABL/c mices, in 6～8 weeks, male and female half and half are divided into matched group i.e. promptly second group of first group and experimental group, 10 every group at random:
First group: the content of Quercetin is 5mg/ml in the Quercetin conventional liposome, every mice dosage 0.2ml, and the tail intravenously administrable is once.
Got eyeball blood after every mice administration in 5 minutes, 15 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes, anticoagulant heparin, centrifuging and taking blood plasma, simultaneously with its viscera tissue after one's own heart, lung, liver,spleen,kidney take out respectively, after the homogenate, extraction detects the concentration of Quercetin and metabolite different time thereof and in the distribution of each organ with HPLC.
Second group: the content of Quercetin is 5mg/ml in the quercetin long-acting liposome, every mice dosage 0.2ml, and the tail intravenously administrable is once.
Got eyeball blood after every mice administration in 5 minutes, 15 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes, 300 minutes, 360 minutes, 480 minutes, 720 minutes, 1440 minutes, anticoagulant heparin, centrifuging and taking blood plasma, simultaneously with its viscera tissue after one's own heart, lung, liver,spleen,kidney take out respectively, after the homogenate, extraction detects the concentration of Quercetin and metabolite different time thereof and in the distribution of each organ with HPLC.
Adopt HPLC to detect mice blood drug level result as can be seen, 15 minutes its blood drug level of Quercetin conventional liposome group mice administration reaches peak.4 hours its blood drug level of quercetin long-acting liposome group mice administration reaches peak.The quercetin long-acting liposome that promptly adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE has significantly prolonged medicine at the intravital circulation time of mice.And after each organ detected and finds injection quercetin long-acting liposome powder for injection, it is the highest at the Quercetin and the metabolite concentration thereof of liver, lung that the same time is got tissue detection.Compartment model adopts two-compartment model.
The measurement result of blood drug level in the table 4 mice body
| Quercetin conventional liposome injectable powder | Time (min) | Concentration C (mg/L) | In(C) | T1/2α (min) | T1/2β (min) | AUC(0-t) (mg/L*min) | AUC(0-∞) (mg/L*min) | Cmax(mg/L) |
| 5 | 10.37 | 2.339 | 35.259 | 69.315 | 1090.142 | 1404.176 | 17.39 | |
| 15 | 17.39 | 2.856 | ||||||
| 30 | 7 | 1.946 | ||||||
| 60 | 4.3 | 1.459 | ||||||
| 120 | 2.95 | 1.082 | ||||||
| 180 | 2.57 | 0.944 | ||||||
| 240 | 1.35 | 0.3 | ||||||
| Quercetin long-acting liposome powder for injection | 5 | 4.85 | 1.579 | 69.315 | 69.315 | 13415.599 | 18993.337 | 20.39 |
| 15 | 3.53 | 1.261 | ||||||
| 30 | 3.32 | 1.2 | ||||||
| 60 | 5.84 | 1.765 | ||||||
| 120 | 12.62 | 2.535 | ||||||
| 180 | 15.62 | 2.749 | ||||||
| 240 | 20.39 | 3.015 | ||||||
| 300 | 17.3 | 2.851 | ||||||
| 360 | 13.2 | 2.58 | ||||||
| 480 | 8.26 | 2.111 | ||||||
| 720 | 5.23 | 1.654 | ||||||
| 1440 | 1.625 | 0.486 |
Pharmacological testing of the present invention and result:
2. get 40 of BABL/c mices, in 6～8 weeks, male and female half and half are divided into 5 groups at random, and first group and second group is matched group, and the 3rd group, the 4th group and the 5th group is experimental group, in mice right fore oxter plantation 5 * 10
5Individual C26 colon cancer cell line, when tumor was grown to 0.5 * 0.5cm size, it is as follows respectively that each organizes pharmacological evaluation:
First group: the normal saline group, 2 tail veins are given the 0.2ml normal saline weekly.
Second group: liposome group, liposome concentration are 5mg/ml, every dosage 0.2ml, 2 tail intravenously administrables weekly.
The 3rd group: the content of Quercetin is 0.05mg/ml in the quercetin long-acting liposome, every dosage 0.2ml, 2 tail intravenously administrables weekly.
The 4th group: the content of Quercetin is 0.5mg/ml in the quercetin long-acting liposome, every dosage 0.2ml, 2 tail intravenously administrables weekly.
The 5th group: the content of Quercetin is 5mg/ml in the quercetin long-acting liposome, every dosage 0.2ml, 2 tail intravenously administrables weekly.
Below respectively organize 4 weeks of successive administration, survey the size of tumor weekly, statistical data, Quercetin concentration is that the therapeutic effect of 0.05mg/ml dosage group is the most obvious in the treatment group, and increased dosage amount it change not quite, and it suppresses treatment group contrast normal saline group tumor rate and reaches 50%.Body weight, hair, action and the feeding treatment group of observing mice in therapeutic process are all good than normal saline matched group, and liver, lung, kidney, brain, heart tissue that mice is organized in treatment are detected, and all do not have the pathological changes reaction.
From above result, it is as follows further to draw beneficial effect of the present invention:
The present invention wraps up the Quercetin of biologically active with the long-acting liposome carrier, be prepared into quercetin long-acting liposome powder for injection, in preparation process, improve raw materials used formula proportion and optimize preparation parameter, the water solublity and the stability of Quercetin have been strengthened, and in the composition of preparation Quercetin liposome, add a certain proportion of Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, its effect is to have stoped plasma protein to be adsorbed in the surface of liposome, as there is not a Quercetin liposome of Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, plasma protein adheres on the surface of liposome very soon, has excited mononuclear phagocyte system to the quick removing of liposome from blood circulation.The liposome that contains Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE can stop adsorption largely, prolong blood circulation time, so drug-loaded liposome can reach diseased region effectively.Cancer increases position and infection, inflammation part in vivo, and pathological changes causes that blood capillary ground permeability increases, the long-acting liposome that contains Quercetin can be increased in the aggregate amount at these positions, and its complete capillary tube of normal structure makes not porous of most liposome.At the Quercetin liposome of diseased region, because the slow release of medicine directly acts on diseased region, increased therapeutic effect, be the active targeting of having realized medicine, prolonged Quercetin circulation time in vivo, improved the bioavailability of Quercetin.
The present invention prepares the used key instrument of quercetin long-acting liposome powder for injection: Rotary Evaporators, Ultrasonic Cell Disruptor, vacuum desiccator, freeze dryer.
Embodiment 1
With 60mg lecithin, the 6mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 12mg, the molecular mass of Polyethylene Glycol is 2000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the Quercetin of 20mg is dissolved in the 100ml organic solvent of chloroform and methanol, chloroform 75ml wherein, methanol 25ml, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, consoluet solution is placed the flask of 250ml, flask is linked to each other with Rotary Evaporators, in the water bath with thermostatic control that the flask intrusion is 40 ℃, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 100ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, Ultrasound Instrument power is 200W, ultrasonic time is 1 hour, the sorbitol of 1mg is dissolved in the quercetin long-acting liposome solution, it is positioned over forms lyophilized powder in the freeze dryer again, be the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 2
With 0.50g lecithin, 0.10g cholesterol, 0.09g Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the molecular weight of Polyethylene Glycol is 3000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, 0.30g Quercetin be dissolved in the 1L organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=1: 1, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, consoluet solution is placed the eggplant type flask of 2.5L, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 30 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even lipid membrane, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 1L, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 800W, ultrasonic time is 2 hours, the sorbitol of 0.005g is dissolved in the quercetin long-acting liposome solution, again it is positioned over and forms lyophilized powder in the freeze dryer, be the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 3
With 45mg lecithin, the 8mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 8mg, the molecular weight of Polyethylene Glycol is 2000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the 35mg Quercetin is dissolved in the organic solvent of chloroform and methanol 200ml, the volume ratio of chloroform and methanol is v: v=4: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 500ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 35 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 200ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 600W, ultrasonic time is 1 hour, mannitol with 4mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer then, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 4
With 39.9mg lecithin, the 5mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 5mg, the molecular weight of Polyethylene Glycol is 3000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the 50mg Quercetin is dissolved in the organic solvent of chloroform and methanol 250ml, the volume ratio of chloroform and methanol is v: v=2: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 500ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 50 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 250ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 400W, ultrasonic time is 1 hour, the sucrose dissolved of 5mg in quercetin long-acting liposome solution, is positioned over it and forms lyophilized powder in the freeze dryer, be the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 5
With 40mg lecithin, the 10mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 12mg, the molecular weight of Polyethylene Glycol is 4000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the Quercetin of 35mg is dissolved in the 150ml organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=3: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 250ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 45 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film,. put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 150ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 400W, ultrasonic time is 1.5 hours, the glucose of 1mg is dissolved in the quercetin long-acting liposome solution, it is positioned over forms lyophilized powder in the freeze dryer again, be the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 6
With 0.60g lecithin, 0.08g cholesterol, 0.10g Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the molecular weight of Polyethylene Glycol is 5000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, 0.20g Quercetin be dissolved in the 1L organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=1: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 2.5L, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 40 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 1L, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 600W, ultrasonic time is 2 hours, trehalose with 5mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 7
With 40mg lecithin, the 5mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 5mg, the molecular weight of Polyethylene Glycol is 4000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, 49.5mg Quercetin is dissolved in the organic solvent of chloroform and methanol 200ml, the volume ratio of chloroform and methanol is v: v=4: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 500ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 45 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 200ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 600W, ultrasonic time is 1 hour, mannitol with 2mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 8
With 45mg lecithin, the 9mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 5mg, the molecular weight of Polyethylene Glycol is 6000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the 40mg Quercetin is dissolved in the organic solvent of chloroform and methanol 300ml, the volume ratio of chloroform and methanol is v: v=2: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 500ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 35 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 300ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 800W, ultrasonic time is 1 hour, trehalose with 1mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 9
With 55mg lecithin, the 10mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 5mg, the molecular weight of Polyethylene Glycol is 8000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the Quercetin of 25mg is dissolved in the 100ml organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=3: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 100ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 40 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 100ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 200W, ultrasonic time is 2 hours, again with the sucrose dissolved of 4mg in quercetin long-acting liposome solution, it is positioned over forms lyophilized powder in the freeze dryer again, be the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
With 40mg lecithin, the 10mg cholesterol, 14.8mg Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the molecular weight of Polyethylene Glycol is 7000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the Quercetin of 35mg is dissolved in the 200ml organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=4: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 500ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 40 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 200ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 700W, ultrasonic time is 1 hour, glucose with 3mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 11
With 0.40g lecithin, 0.05g cholesterol, 0.05g Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the molecular weight of Polyethylene Glycol is 8000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, 0.47g Quercetin be dissolved in the 1L organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=1: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, this dissolved solution is placed the eggplant type flask of 2.5L, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 50 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 1L, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 800W, ultrasonic time is 2 hours, trehalose with 0.001g is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Embodiment 12
With 39mg lecithin, the 5mg cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE of 5mg, the molecular weight of Polyethylene Glycol is 8000 in Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, the Quercetin of 50mg is dissolved in the 150ml organic solvent of chloroform and methanol, the volume ratio of chloroform and methanol is v: v=2: 1, volume is with milliliter ml or rise the L metering, in airtight bottle, be stirred to lecithin, cholesterol, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE and Quercetin dissolve fully, dissolved solution is placed the eggplant type flask of 250ml, eggplant type flask is linked to each other with rotary evaporator, flask is invaded 30 ℃ water bath with thermostatic control, the evacuation rotary evaporation is removed organic solvent, on flask walls, form the layer of even thin film, put it in the vacuum drying oven then, dry 2 hours at ambient temperature, the aquesterilisa that adds 150ml, carry out ultra-sonic dispersion with Ultrasonic Cell Disruptor, ultrasonic power is 400W, ultrasonic time is 1 hour, sorbitol with 5mg is dissolved in the quercetin long-acting liposome solution again, it is positioned over forms lyophilized powder in the freeze dryer again, is the cotton-shaped Quercetin liposome powder for injection of yellow green.With its evacuation or charge into inert gas seal, be positioned over temperature and be preserve under 4 ℃ of-8 ℃ of conditions standby.
Claims (14)
1. quercetin long-acting liposome powder for injection, it is characterized in that it is made up of Quercetin 20-50 part, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE 5-15 part, lecithin 40-60 part, cholesterol 5-15 part and excipient 0.1-5 part of biologically active, wherein the molecular mass of Polyethylene Glycol is 2000-8000, the consumption proportion of lecithin and cholesterol is 3-12: 1, its component all by weight, described excipient is selected from sorbitol, or mannitol, or glucose, or sucrose, or trehalose.
2. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 2000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
3. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 2000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 10-15 part.
4. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 4000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
5. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 4000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-11 part.
6. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 8000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
7. according to the described injectable powder of claim 1, the molecular weight that it is characterized in that used Polyethylene Glycol is 8000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 8-10 part.
8. the preparation method of arbitrary described quercetin long-acting liposome powder for injection among the claim 1-7, adopt the thin-film ultrasonic method, it is characterized in that: adopt the Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE modification lecithin of different molecular quality and the surperficial long-acting liposome that gets of conventional liposome of cholesterol composition, the Quercetin of long-acting liposome parcel biologically active, this method is raw materials used and consumption, technological parameter and processing step are as follows:
(1) Quercetin 20-50 part, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE 5-15 part, lecithin 40-60 part, cholesterol 5-15 part, wherein the proportioning of lecithin and cholesterol is 3-12: 1, the molecular mass of used Polyethylene Glycol is 2000-8000, above raw material is solid-state, and with milligram or gram or kilogram metering, organic solvent chloroform and methanol are liquid, their volume proportion is 1-4: 1, with milliliter or liter metering;
Quercetin, Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE, lecithin, the cholesterol that measures is dissolved in the chloroform and methanol organic solvent that proportioning is good according to the above ratio, making the gained solution concentration is 1mg/ml-3mg/ml, place then in the airtight bottle, be stirred to dissolving fully at ambient temperature;
(2) with in the flask that above-mentioned consoluet solution is transferred to rotary evaporator is connected, it is that the evacuation rotary evaporation is removed organic solvent then, forms the layer of even thin film to flask walls in 30 ℃-50 ℃ the water bath with thermostatic control that flask is invaded temperature;
(3) flask that will be formed with thin film is put into vacuum drying oven, dry 2 hours at ambient temperature, further removes residual organic solvent;
(4) aquesterilisa is added in the dried flask, make the complete submergence of thin film of bottle wall, use the Ultrasonic Cell Disruptor ultra-sonic dispersion, its ultrasonic power is 100w-1000w, and ultrasonic time is 0.5-3 hour, promptly gets quercetin long-acting liposome solution;
(5) add excipient in quercetin long-acting liposome solution, after the lyophilizing, the cryogenic conditions lower cover sealing preservation of depositing in 4 ℃-8 ℃ is standby, and excipient is selected from sorbitol, or mannitol, or glucose, or sucrose, or trehalose.
9. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 2000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
10. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 2000 o'clock, and the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 10-15 part.
11. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 4000 o'clock, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
12. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 4000 o'clock, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-11 part.
13. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 8000 o'clock, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 5-15 part.
14. according to the described injectable powder preparation method of claim 8, the molecular weight that it is characterized in that used Polyethylene Glycol is 8000 o'clock, the amount that adds Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE is 8-10 part.
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| CN102058536A (en) * | 2011-01-14 | 2011-05-18 | 四川大学 | Quercetin hydroxypropyl Beta-cyclodextrin inclusion liposome, preparation method and application thereof |
| CN103070826B (en) * | 2012-12-31 | 2015-09-02 | 清华大学深圳研究生院 | A kind of Quercetin Dermatological Liposomes and lyophilized powder and its production and use thereof |
| UA111762C2 (en) * | 2014-07-08 | 2016-06-10 | ТОВАРИСТВО З ОБМЕЖЕНОЮ ВІДПОВІДАЛЬНІСТЮ "НаноМедТраст" | A method of obtaining a pharmacologically active liposomal drug containing quercetin |
| CN106236711A (en) * | 2016-08-25 | 2016-12-21 | 广东工业大学 | Phloretin liposome and preparation method thereof |
| US11331298B2 (en) | 2017-07-17 | 2022-05-17 | Indena S.P.A. | Powder solid dispersions comprising quercetin, process for their preparation and formulations thereof |
| CN108042492B (en) * | 2017-12-18 | 2020-06-16 | 成都大学 | Tartary buckwheat flavone lipid polymer nanoparticles and preparation method thereof |
| CN109507324A (en) * | 2018-12-11 | 2019-03-22 | 重庆医科大学 | The detection method of quercetin content in a kind of measurement liver tissues of rats |
| CN114010651A (en) * | 2021-12-03 | 2022-02-08 | 南通大学 | Improved isoquercitrin and preparation method and application thereof |
| CN116440081A (en) * | 2023-04-03 | 2023-07-18 | 上海拜思丽实业有限公司 | Preparation method of quercetin liposome |
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