CN103610639B - Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt - Google Patents
Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt Download PDFInfo
- Publication number
- CN103610639B CN103610639B CN201310606778.2A CN201310606778A CN103610639B CN 103610639 B CN103610639 B CN 103610639B CN 201310606778 A CN201310606778 A CN 201310606778A CN 103610639 B CN103610639 B CN 103610639B
- Authority
- CN
- China
- Prior art keywords
- buffer salt
- polymer micelle
- medicine carrying
- salt solution
- polymer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The invention discloses a kind of polymer micelle medicine carrying compositions containing buffer salt, comprising: amphipathic nature block polymer, hydrophobic drug and buffer salt solution; The mass ratio of described amphipathic nature block polymer and described hydrophobic drug is 99:1 ~ 4:1.Polymer micelle medicine carrying composition and method of making the same containing buffer salt provided by the present invention, adding by buffer salt solution, the physical stability of polymer micelle solution can be significantly improved, obviously can reduce the content of hydrophobic drug related substance, improve the quality of preparation, thus meet the requirement of suitability for industrialized production and clinical application time.
Description
Technical field
The present invention relates to a kind of polymer micelle medicine carrying composition and method of making the same, particularly relate to a kind of polymer micelle medicine carrying composition and method of making the same containing buffer salt, belong to chemical pharmaceutical technical field.
Background technology
Bearing taxanes is the large class natural product obtained from plant, belongs to Diterpenes from textural classification, therefore also known as Taxane diterpenes.Paclitaxel, as the star molecule in this compounds, is a kind of important anticarcinogen used clinically.Paclitaxel is a kind of natural anti-cancer crude drug extracted from Ramulus et folium taxi cuspidatae or Chinese yew, is white crystalline powder, water insoluble, easily dissolves in chloroform, acetone and other organic solvent.Docetaxel is the compound being synthesized preparation by yew needle extract through structural modification, for white or off-white color powdery solid, also taxane anti-tumor medicament is belonged to, for fat-soluble compound, be insoluble in water (2.903 μ g/mL), slightly larger than paclitaxel (0.25 μ g/mL ~ 0.60 μ g/mL).This product antitumor mechanism is identical with paclitaxel, is also by inducing and impelling tubulin polymerization to become microtubule, suppress simultaneously form the depolymerization of microtubule, produce stable microtubule fasolculus.The normal dynamic of microtubule fasolculus is regenerated be obstructed, cell can not form normal mitosis spindle when mitosis, thus inhibits cell division and propagation.Its unique anti-tumor mechanism of action has unique therapeutical effect to breast carcinoma, ovarian cancer and non-small cell carcinoma etc., the extensive use as a line anticarcinogen clinically.But as other many anticarcinogen, the dissolubility of taxone in water is all very low, in the mixed solution of the every mL of paclitaxel injection used clinically at present containing Oleum Ricini and 49.7% (V/V) dehydrated alcohol of the rare modification of 6mg paclitaxel 527mg polyoxy second, use normal saline dilution 5-20 before injection doubly.The solution that docetaxel injection is also made up of Tween 80 and the ethanol of docetaxel, clinic trial is somewhat similar with paclitaxel injection; But
eL, Tween 80 and ethanol often cause strong side reaction, as allergy, haemolysis, nephrotoxicity, neurotoxicity and cardiac toxicity etc., may cause death time serious.And increase with dosage, toxic and side effects has the trend of increase.Therefore strongly limit the clinical practice of the taxones such as paclitaxel.
At present, existing large quantifier elimination shows that polymer micelle system can be used as the carrier of the medicine being insoluble in water.The feature of polymer micelle is, in water-bearing media, there is a nucleocapsid structure, and it derives from the amphiphilic diblock copolymer with hydrophobic (core) and hydrophilic (shell) part.Polymer micelle medicine carrying is that the medicine owing to being insoluble in water is similar to the polarity of hydrophobic core part, thus the pharmaceutical pack being insoluble in water is embedded in the hydrophobic core of micelle.The body circulation time long due to polymer micelle and the passive target effect by EPR, they change the drug distribution after by intravenous administration, because this reducing side effect and improve its effect, thus the advantage provided as cell-specific targeting control the release of medicine.Using biodegradable polymer micelle system to be in the news as the carrier of the medicine being insoluble in water is very useful technology.
To have in prior art and use block copolymer, for A-B type diblock copolymer, it comprises as the polyethylene glycol oxide of hydrophilic component A or polylalkylene glycol derivatives with as the polyamino acid with hydrophobic functional groups of hydrophobicity B component or its derivant, for physically the medicine of such as amycin, indomethacin is incorporated in the core of polymer micelle, is then used as the carrier of drug release.But these polymer micelles are made up of the block copolymer of in vivo not easily degrading.In addition, this block copolymer has low biocompatibility, can cause undesirable side effect (as: immunoreation etc.) in vivo when it uses.
In prior art, multiple preparation method all can realize the preparation of nano-micelle, but the micelle physical stability that the amphipathic copolymer prepared by its formulation and technology wraps up hydrophobic drug can only reach several tens minutes to a few hours, and stability is bad.
Summary of the invention
For the deficiency existing for prior art, technical problem to be solved by this invention is to provide a kind of polymer micelle medicine carrying composition and method of making the same containing buffer salt.Said composition and method significantly can improve the physical stability of polymer micelle medicine carrying compositions, have good biocompatibility, thus meet the requirement of suitability for industrialized production and clinical application time.
For realizing above-mentioned object, the present invention adopts following technical scheme:
On the one hand, the invention provides a kind of polymer micelle medicine carrying compositions containing buffer salt, it is characterized in that comprising:
Amphipathic nature block polymer, hydrophobic drug and buffer salt solution;
The mass ratio of described amphipathic nature block polymer and described hydrophobic drug is 99:1 ~ 4:1.
Wherein more preferably, the mass ratio of described Amphipathilic block polymer and described hydrophobic drug is 19:1.
Wherein more preferably, described amphipathic nature block polymer is selected from the one in mono methoxy polyethylene glycol-PLA, mono methoxy polyethylene glycol-polycaprolactone, PCL-PEG-PCL.
Wherein more preferably, described amphipathic nature block polymer be selected from molecular weight be 3600 ~ 4400 mono methoxy polyethylene glycol-PLA, molecular weight be 4000 mono methoxy polyethylene glycol-polycaprolactone, molecular weight be one in the PCL-PEG-PCL of 3700.
Wherein more preferably, the concentration of described hydrophobic drug is 0.1 ~ 10mg/mL.
Wherein more preferably, the concentration of described hydrophobic drug is 0.4 ~ 4mg/mL.
Wherein more preferably, the concentration of described hydrophobic drug is 1mg/mL.
Wherein more preferably, described hydrophobic drug is selected from bearing taxanes.
Wherein more preferably, the pH of described buffer salt solution is 6 ~ 10.
On the other hand, the present invention also provides a kind of preparation method of above-mentioned polymer micelle medicine carrying compositions, it is characterized in that comprising the steps:
By hydrophobic drug and amphipathic nature block polymer organic solvent dissolution;
Polymer micelle is obtained after concentrating under reduced pressure removing organic solvent;
Add buffer salt solution dispersion or first add buffer salt solution dispersion again with after water for injection dispersion, obtaining polymer micelle medicine carrying compositions.
Wherein more preferably, described organic solvent is the one in ethanol, acetone, acetonitrile or dichloromethane.
Wherein more preferably, described amphipathic nature block polymer is prepared as follows:
Mono methoxy polyethylene glycol concentrating under reduced pressure removing moisture;
Mix after adding catalyst;
Add lactide mixing to heat up;
Cool and be dissolved in dehydrated alcohol;
After injecting pentane, purification is dry obtains amphipathic nature block polymer.
Wherein more preferably, described amphipathic nature block polymer is mono methoxy polyethylene glycol-poly-(D, L-lactide) (mPEG-PDLLA), mono methoxy polyethylene glycol-polycaprolactone (mPEG-PCL), PCL-PEG-PCL (PCEC), poly-(D, L-lactide) one in-polyethylene glycol-(D, L-lactide).
Wherein more preferably, described buffer salt solution is selected from the one in sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution, dipotassium hydrogen phosphate/potassium dihydrogen phosphate buffer salt solution, sodium hydrogen phosphate/citric acid buffer salt solution, sodium bicarbonate buffer saline solution or sodium bicarbonate/sodium carbonate buffer salt solution
Wherein more preferably, the pH of described buffer salt solution is 7.8 ~ 9.6.
Polymer micelle medicine carrying composition and method of making the same containing buffer salt provided by the present invention, adding by buffer salt solution, can significantly improve the physical stability of polymer micelle solution, thus meet the requirement of suitability for industrialized production and clinical application time.By first adding buffer salt solution again with after water for injection dispersion, obviously can reduce the content of hydrophobic drug related substance, improving the quality of preparation.
Accompanying drawing explanation
Fig. 1 is the NMR collection of illustrative plates of mPEG-PDLLA.
Fig. 2 is the drug leakage result of the test figure of different aqueous vehicles.
In Fig. 3 embodiment seven, dispersing mode one prepares the chromatogram of polymer micelle solution.
In Fig. 4 embodiment seven, dispersing mode two prepares the chromatogram of polymer micelle solution.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail.
The invention provides a kind of polymer micelle medicine carrying compositions containing buffer salt, comprising: amphipathic nature block polymer, hydrophobic drug and buffer salt solution; The mass ratio of amphipathic nature block polymer and hydrophobic drug is 99:1 ~ 4:1.Prove by experiment, best when the mass ratio of Amphipathilic block polymer and hydrophobic drug is 19:1.Detailed description is launched to this polymer micelle medicine carrying compositions below.
In an embodiment of the present invention, easily degrade in human body to make the polymer micelle medicine carrying compositions of loaded with anti-tumor medicine, amphipathic nature block polymer is selected from mono methoxy polyethylene glycol-poly-(D, L-lactide) (mPEG-PDLLA), mono methoxy polyethylene glycol-polycaprolactone (mPEG-PCL), PCL-PEG-PCL (PCEC) or poly-(D, L-lactide)-polyethylene glycol-(D, L-lactide).Above-mentioned amphipathic nature block polymer has good biocompatibility, avoids causing the side effect such as immunoreation in vivo.When mono methoxy polyethylene glycol-polycaprolactone that mono methoxy polyethylene glycol-PLA that amphipathic nature block polymer preferred molecular weight is 3600 ~ 4400, molecular weight are 4000, molecular weight are a kind of in the PCL-PEG-PCL of 3700, polymer micelle medicine carrying compositions stability is better.
Learn from experimental verification, pH is polymer micelle medicine carrying compositions stabilization time very long (can reach more than 18h) prepared of buffer salt solution of 6 ~ 10, and pH is respectively the polymer micelle medicine carrying compositions steady time the longest (for up to 36h) prepared by 7.8 ~ 9.6 buffer salt solutions.In order to make the polymer micelle medicine carrying compositions of loaded with anti-tumor medicine longer for stabilization time, buffer salt solution is selected from the one in sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution, dipotassium hydrogen phosphate/potassium dihydrogen phosphate buffer salt solution, sodium hydrogen phosphate/citric acid buffer salt solution, sodium bicarbonate buffer saline solution or sodium bicarbonate/sodium carbonate buffer salt solution; The concentration of described hydrophobic drug is 0.1 ~ 10mg/mL.The concentration of hydrophobic drug is preferably 1mg/mL.
Hydrophobic drug is preferably from bearing taxanes.Hydrophobic drug preferably selects docetaxel or paclitaxel.
The present invention also provides a kind of preparation method of above-mentioned polymer micelle medicine carrying compositions, comprises the steps: hydrophobic drug and amphipathic nature block polymer organic solvent dissolution; Polymer micelle is obtained after concentrating under reduced pressure removing organic solvent; Add buffer salt solution dispersion or first add buffer salt solution dispersion again with after water for injection dispersion, obtaining polymer micelle medicine carrying compositions.
In embodiments of the invention, when preparing polymer micelle medicine carrying compositions, organic solvent can select the one in ethanol, acetone, acetonitrile or dichloromethane.Amphipathic nature block polymer is preferably prepared as follows: mono methoxy polyethylene glycol concentrating under reduced pressure removing moisture; Mix after adding catalyst; Add lactide mixing to heat up; Cool and be dissolved in dehydrated alcohol; After injecting pentane, purification is dry obtains amphipathic nature block polymer.
Below in conjunction with multiple embodiment, detailed description is launched to the polymer micelle medicine carrying composition and method of making the same containing buffer salt provided by the invention.
The preparation of embodiment one mPEG-PDLLA copolymer
10g mono methoxy polyethylene glycol (mPEG, molecular weight is 2000) is joined in 100mL tri-neck round-bottomed flask, in oil bath pan, is heated to 100 DEG C, be reduced to the pressure of 500Pa by vacuum aspirator under, carry out 30min to remove unnecessary moisture content.Add the catalyst of stannous octoate as reaction of 61.65mg, stirring and evenly mixing 30min under nitrogen atmosphere.In flask, add the lactide of 11g again, mixing, is warming up to 130 DEG C, stirring reaction 18h.The product obtained is cooled to 50 DEG C and is dissolved in dehydrated alcohol.This solution is injected into (-20 DEG C) in cold pentane, the diblock copolymer be precipitated.By dissolving-process of precipitation repeats twice, makes mPEG-PDLLA diblock copolymer (MW:2000-2000 dalton) obtain purification, vacuum drying and get final product.The NMR(D-chloroform of product is solvent) collection of illustrative plates confirms the quality when molecular weight of methyl Polyethylene Glycol in polymer and lactide with b peak (PEG) and c peak (PDLLA) peak area ratio, result is as shown in Figure 1.
Embodiment two buffer salt solution is on the impact of polymer micelle medicine carrying compositions stability
Get 2mL190mg/mLmPEG-PDLLA alcoholic solution in round-bottomed flask, add 2mL10mg/mL docetaxel alcoholic solution, by round-bottomed flask as on Rotary Evaporators, decompression (0.08MPa), water-bath 30 DEG C, rotating speed 100r/min, until removing ethanol, takes off round-bottomed flask, add 20mL water for injection, 60 DEG C of water-bath concussion 3min, gained solution, through 0.22 μm of membrane filtration, is got filtrate and is loaded in 10mL cillin bottle.
Water for injection is replaced as buffer salt solution respectively using sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8), sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH8.0), dipotassium hydrogen phosphate/potassium dihydrogen phosphate buffer salt solution (pH7.8), sodium hydrogen phosphate/citric acid buffer salt solution (pH7.8), sodium bicarbonate buffer saline solution (pH8.0), sodium bicarbonate/sodium carbonate buffer salt solution (pH9.6), prepare as stated above, obtain buffer salt-polymer micelle.The stability of 25 DEG C of observation polymer micelle medicine carrying compositionss, the results are shown in Table 1.Stabilization time is after adding aqueous vehicles until the time having medicine crystal to separate out.
As known from Table 1, the polymer micelle medicine carrying compositions prepared with the buffer salt solution that pH is 7.8 ~ 9.6 can reach more than 18h stabilization time, sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution that pH is respectively 7.8 and 8.0 reaches 36h as stabilization time during bunchy medium, and stabilization time when not adding buffer salt solution is only 1h, show that the stability of the polymer micelle medicine carrying compositions adding buffer salt improves greatly.
| Bunchy medium | Stabilization time |
| Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8) | 36h |
| Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH8.0) | 36h |
| Dipotassium hydrogen phosphate/potassium dihydrogen phosphate buffer salt solution (pH7.8) | 24h |
| Sodium hydrogen phosphate/citric acid buffer salt solution (pH7.8) | 18h |
| Sodium bicarbonate buffer saline solution (pH8.0) | 30h |
| Sodium bicarbonate/sodium carbonate buffer salt solution (pH9.6) | 22h |
| Water for injection | 1h |
Table 1 adds buffer salt and compares with the test not adding buffer salt
Note: in table 1, the compound method of buffer salt solution is as follows:
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 91.5mL to mix with second liquid 8.5mL, shake up, to obtain final product.
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 94.7mL to mix with second liquid 5.3mL, shake up, to obtain final product.
Dipotassium hydrogen phosphate/potassium dihydrogen phosphate buffer salt solution (pH7.8): get dipotassium hydrogen phosphate 5.59g and potassium dihydrogen phosphate 0.41g, add water and make to be dissolved into 1000mL, to obtain final product.
Sodium hydrogen phosphate/citric acid buffer salt solution (pH7.8): first liquid: get citric acid 21g or anhydrous citric acid 19.2g, be dissolved in water into 1000mL, put Refrigerator store.Second liquid: get sodium hydrogen phosphate 71.63g, adds water and makes to be dissolved into 1000mL, gets above-mentioned first liquid 95.75mL and mixes with second liquid 4.25mL, shake up, to obtain final product.
Sodium bicarbonate buffer saline solution (pH8.0): get sodium bicarbonate 5g, be dissolved in water, and be diluted to 100mL, shake up, to obtain final product.
Sodium bicarbonate/sodium carbonate buffer salt solution (pH9.6): get natrium carbonicum calcinatum 63.6g and sodium bicarbonate 33.6g, be dissolved in water, and be diluted to 1000mL, shake up, to obtain final product.
Embodiment three pH value is on the impact of polymer micelle medicine carrying compositions stability
Get 2mL190mg/mLmPEG-PDLLA alcoholic solution in round-bottomed flask, add 2mL10mg/mL docetaxel alcoholic solution, by round-bottomed flask as on Rotary Evaporators, decompression (0.08Mpa), water-bath 30 DEG C, rotating speed 100r/min, until removing ethanol, take off round-bottomed flask, add 20mL buffer salt solution, 60 DEG C of water-bath concussion 3min, to clarification, gained solution, through 0.22 μm of membrane filtration, is got 20mL and is loaded in cillin bottle, observes stability.
Sodium hydrogen phosphate/sodium dihydrogen phosphate the buffer salt solution improving stabilizing effect the best is made into respectively the buffer salt solution of different pH value, prepares as stated above, obtain buffer salt-polymer micelle.The stability of 25 DEG C of observation polymer micelle medicine carrying compositionss, the results are shown in Table 2.Stabilization time is after adding aqueous vehicles until the time having medicine crystal to separate out.As known from Table 2, when the pH of sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution is 7.8, stabilization time is the longest.
| Docetaxel amount | Copolymer amount | Buffer salt solution pH | Stabilization time |
| 20mg | 380mg | 5.8 | 1h |
| 20mg | 380mg | 7.8 | 36h |
| 20mg | 380mg | 8.0 | 35h |
| 20mg | 380mg | 6.0 | 4h |
| 20mg | 380mg | 9.6 | 20h |
Polymer latex beam stability prepared by the different pH buffer salt solution of table 2
Note: in table 2, the compound method of buffer salt solution is as follows:
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH5.8): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 8mL to mix with second liquid 92mL, shake up, to obtain final product.
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 91.5mL to mix with second liquid 8.5mL, shake up, to obtain final product.
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH8.0): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 94.7mL to mix with second liquid 5.3mL, shake up, to obtain final product.
Sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH6.0): first liquid: get sodium hydrogen phosphate 35.9g, be dissolved in water, and be diluted to 500mL.Second liquid: get sodium dihydrogen phosphate 2.76g, be dissolved in water, and be diluted to 100mL.Get above-mentioned first liquid 12.3mL to mix with second liquid 87.7mL, shake up, to obtain final product.
Sodium bicarbonate/sodium carbonate buffer salt solution (pH9.6): get natrium carbonicum calcinatum 63.6g and sodium bicarbonate 33.6g, be dissolved in water, and be diluted to 1000mL, shake up, to obtain final product.
The impact of embodiment four polymer on polymer micelle medicine carrying compositions stability
By the preparation method in embodiment three, adopt the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8 as buffer salt solution, respectively with mono methoxy polyethylene glycol-poly-(D, L-lactide) (mPEG-PDLLA), mono methoxy polyethylene glycol-polycaprolactone (mPEG-PCL) and PCL-PEG-PCL (PCEC) be as polymer, prepare the polymer micelle medicine carrying compositions of docetaxel, the stability of 25 DEG C of observation polymer micelle medicine carrying compositionss, the results are shown in Table 3.As known from Table 3, adopt molecular weight be 3600 ~ 4400 mPEG-PDLLA or molecular weight be 4000 mPEG-PCL or molecular weight be 3700 PCEC as polymer time, micelle stability is better, and particle diameter is all less than 100nm, and PDI is all less than 0.25.
| Copolymer | Molecular weight | Stabilization time | Particle diameter (nm) | PDI |
| mPEG 2000-PDLLA 1350 | 3350 | 5h | 22 | 0.127 |
| mPEG 2000-PDLLA 1600 | 3600 | 24h | 24 | 0.079 |
| mPEG 2000-PDLLA 2050 | 4050 | 24h | 23 | 0.154 |
| mPEG 2000-PDLLA 2400 | 4400 | 18h | 29 | 0.137 |
| mPEG 2000-PCL 2000 | 4000 | 20h | 30 | 0.215 |
| PCL 850-PEG 2000-PCL 850 | 3700 | 22h | 31 | 0.227 |
Polymer latex beam stability, particle diameter and dispersion index prepared by table 3 different copolymer thing
The impact that embodiment five polymer on polymer micelle medicine carrying compositions drug loading, envelop rate, medicine are revealed
By the preparation method in embodiment three, adopt the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8 as buffer salt solution, prepare docetaxel polymer micelle dried frozen aquatic products, add 4mL acetonitrile to dissolve, vortex mixes, and 0.22 μm of membrane filtration, gets filtrate injection liquid chromatography, record chromatogram, by external standard method with calculated by peak area docetaxel content.Chromatograph of liquid: Agilent1260QuatPumpVL, Agilent1260 column oven automatic sampler SL, Agilent1260 column oven, Agilent1260DADVL; Chromatographic column: Kromasil100-3.5C18,150*4.6mm, E69472; Column temperature: 45 DEG C; Wavelength: 232nm; Mobile phase: acetonitrile: water=55:45; Flow velocity: 1mL/min; Sample size: 20 μ L.
By the preparation method in embodiment three, adopt the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8 as buffer salt solution, prepare docetaxel polymer micelle dried frozen aquatic products, add 4mL acetonitrile vortex to dissolve, filter after leaving standstill, sample introduction, record chromatogram, by external standard method with the content of calculated by peak area docetaxel.Docetaxel drug loading, envelop rate are according to following formulae discovery:
Drug loading=content/the dosage of docetaxel (in the micelle of mensuration) × 100%
Envelop rate=(actual drug loading/theoretical drug loading) × 100%
Experimental result is in table 4.As known from Table 4, when the mPEG-PDLLA copolymer of different molecular weight prepares docetaxel polymer micelle, the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution adding pH7.8, as buffer salt solution, can obtain the preparation that drug loading is stablized, envelop rate is high.
The mensuration of table 4 different molecular weight mPEG-PDLLA copolymer drug loading, envelop rate
By the preparation method in embodiment three, use the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8 and water for injection as buffer salt solution respectively, gelatinous polymer micelle after dispersion removing organic solvent, obtained docetaxel polymer micelle solution, place in the environment of constant 25 DEG C, timing sampling 2mL, 0.22 μm of membrane filtration, add 4mL acetonitrile, 0.22 μm of membrane filtration, get filtrate injection liquid chromatography, record chromatogram, by external standard method with the content of calculated by peak area docetaxel, in the drug leakage amount of different time relatively, the results are shown in Figure 2, phosphate buffer in Fig. 2 refers to the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8.As can be seen from Figure 2, the micelle prepared with the sodium hydrogen phosphate of pH7.8/sodium dihydrogen phosphate buffer salt solution and the micellar phase ratio prepared with water for injection, drug leakage speed is slower, release also evenly, the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution describing pH7.8 intuitively can be good at the stability improving preparation.
Embodiment six variable concentrations of hydrophobic medicine is on the impact of polymer micelle medicine carrying compositions stability, particle diameter
Get 2mL190mg/mLmPEG-PDLLA alcoholic solution respectively in round-bottomed flask, add 2mL10mg/mL docetaxel alcoholic solution respectively, by round-bottomed flask as on Rotary Evaporators, decompression (0.08MPa), water-bath 30 DEG C, rotating speed 100r/min, until removing ethanol, take off round-bottomed flask, the phosphate buffered saline (pH7.8) of different volumes is added respectively according to table 5, 60 DEG C of water-bath concussion 3min, gained solution is through 0.22 μm of membrane filtration, getting filtrate is loaded in cillin bottle, investigate variable concentrations of hydrophobic drug solution outward appearance, stability time and particle diameter situation.
The different bunchy concentration of table 5 is on the impact of formulation aesthetics, particle diameter and stability
As known from Table 5, when adding phosphate buffered saline (pH7.8) and preparing the polymer micelle medicine carrying composition solution of hydrophobic drug concentration at 0.1 ~ 10mg/ml, polymer micelle medicine carrying composition solution is clarified, and mean diameter is between 20 ~ 30nm, and stabilization time is in 12 ~ 36h.For producing the filling time of upper abundance, being unlikely to again to cause sublimation drying long simultaneously, the hydrophobic drug concentration of polymer micelle medicine carrying composition solution being controlled at 0.4 ~ 4mg/ml better, preferred 1mg/ml.
The different dispersing mode of embodiment seven is on the impact of preparation stability and related substance
Mode one: get 2mL190mg/mLmPEG-PDLLA alcoholic solution in round-bottomed flask, add 2mL10mg/mL docetaxel alcoholic solution, by round-bottomed flask as on Rotary Evaporators, decompression (0.08MPa), water-bath 30 DEG C, rotating speed 100r/min, until removing ethanol, take off round-bottomed flask, add 20mL sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution (pH7.8), 60 DEG C of water-bath concussion 3min, to clarification, the principal agent concentration of gained solution is 1mg/mL, through 0.22 μm of membrane filtration, getting 20mL is loaded in cillin bottle, lyophilizing immediately obtains docetaxel polymer micelle dried frozen aquatic products.Dried frozen aquatic products adds 0.9% sodium chloride injection or 5% glucose injection 20mL and redissolves the docetaxel nano micellar solution of polymer preparing clear bit bluish opalescence.
Mode two: get 2mL190mg/mLmPEG-PDLLA alcoholic solution in round-bottomed flask, add 2mL10mg/mL docetaxel alcoholic solution, by round-bottomed flask as on Rotary Evaporators, decompression (0.08MPa), water-bath 30 DEG C, rotating speed 100r/min, until removing ethanol, take off round-bottomed flask, add 10mL water for injection, 60 DEG C of water-bath concussion 3min, to clarification, add 10mL phosphate buffered saline (pH7.8) again, jolting or be uniformly mixed, the principal agent concentration of gained solution is 1mg/mL, through 0.22 μm of membrane filtration, getting 20mL is loaded in cillin bottle, lyophilizing immediately obtains docetaxel polymer micelle dried frozen aquatic products.Dried frozen aquatic products adds 0.9% sodium chloride injection or 5% glucose injection 20mL and redissolves the docetaxel nano micellar solution of polymer preparing clear bit bluish opalescence.
Get the docetaxel polymer micelle dried frozen aquatic products prepared according to above-mentioned two kinds of modes respectively, add 4mL acetonitrile and dissolve, vortex mixes, after 0.22 μm of membrane filtration, get filtrate by the liquid phase chromatogram condition sample introduction in embodiment five, record chromatogram, by external standard method with calculated by peak area docetaxel content.Prepare the chromatogram of micellar solution as shown in Figure 3 by mode one, prepare the chromatogram of micellar solution as shown in Figure 4 by mode two.Are compared the stabilization time of two kinds of feed postition and the percentage composition of related substance, result is as shown in table 6.As known from Table 6, the stabilization time impact of different dispersing mode on preparation is little, but pass-through mode two significantly can reduce the percentage composition of docetaxel related substance, is significantly improved to the quality of the pharmaceutical preparations.
| Dispersing mode | Stabilization time | Related substance amount |
| Mode one | 24h | 0.62% |
| Mode two | 24h | 0.29% |
The different dispersing mode of table 6 is on the impact of preparation stability and related substance
In sum, the present invention using pH be the buffer salt solution of 6 ~ 10 as the stabilizing agent of polymer micelle medicine carrying compositions, significantly can improve the physical stability of polymer micelle medicine carrying compositions, thus meet the requirement of suitability for industrialized production and clinical application time; The block copolymer (mPEG-PCL or mPEG-PDLLA or PCEC) that polymer micelle medicine carrying compositions adopts in vivo easily is degraded, and has good biocompatibility, avoids causing the side effect such as immunoreation in vivo.
Above the polymer micelle medicine carrying composition and method of making the same containing buffer salt provided by the present invention is described in detail.For one of ordinary skill in the art, to any apparent change that it does under the prerequisite not deviating from connotation of the present invention, all by formation to infringement of patent right of the present invention, corresponding legal responsibility will be born.
Claims (9)
1., containing a polymer micelle medicine carrying compositions for buffer salt, it is characterized in that comprising:
Amphipathic nature block polymer, hydrophobic drug and buffer salt solution;
Mono methoxy polyethylene glycol-polycaprolactone that described amphipathic nature block polymer is selected from mono methoxy polyethylene glycol-PLA that molecular weight is 3600-4400, molecular weight is 4000, molecular weight are the one in the PCL-PEG-PCL of 3700; The mass ratio of described amphipathic nature block polymer and described hydrophobic drug is 99:1-4:1; Described buffer salt solution is selected from the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8-8.0.
2., as claimed in claim 1 containing the polymer micelle medicine carrying compositions of buffer salt, it is characterized in that the mass ratio of described Amphipathilic block polymer and described hydrophobic drug is 19:1.
3., as claimed in claim 1 containing the polymer micelle medicine carrying compositions of buffer salt, it is characterized in that the concentration of described hydrophobic drug is 0.1-10mg/mL.
4., as claimed in claim 1 containing the polymer micelle medicine carrying compositions of buffer salt, it is characterized in that the concentration of described hydrophobic drug is 0.4-4mg/mL.
5., as claimed in claim 1 containing the polymer micelle medicine carrying compositions of buffer salt, it is characterized in that the concentration of described hydrophobic drug is 1mg/mL.
6., as claimed in claim 1 containing the polymer micelle medicine carrying compositions of buffer salt, it is characterized in that described hydrophobic drug is selected from bearing taxanes.
7. the preparation method of the polymer micelle medicine carrying compositions containing buffer salt as described in one of claim 1-6, is characterized in that comprising the steps:
By hydrophobic drug and amphipathic nature block polymer organic solvent dissolution;
Polymer micelle is obtained after concentrating under reduced pressure removing organic solvent;
Add buffer salt solution dispersion or first add buffer salt solution dispersion again with after water for injection dispersion, obtaining polymer micelle medicine carrying compositions; Described buffer salt solution is selected from the sodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt solution of pH7.8-8.0.
8. the preparation method of polymer micelle medicine carrying compositions as claimed in claim 7, it is characterized in that, described amphipathic nature block polymer is prepared as follows:
Mono methoxy polyethylene glycol concentrating under reduced pressure removing moisture;
Mix after adding catalyst;
Add lactide mixing to heat up;
Cool and be dissolved in dehydrated alcohol;
After injecting pentane, purification is dry obtains amphipathic nature block polymer.
9. the preparation method of polymer micelle medicine carrying compositions as claimed in claim 7, it is characterized in that described amphipathic nature block polymer is selected from mono methoxy polyethylene glycol-PLA that molecular weight is 3600-4400, molecular weight is 4000 mono methoxy polyethylene glycol-polycaprolactone, molecular weight is one in the PCL-PEG-PCL of 3700.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310606778.2A CN103610639B (en) | 2013-10-29 | 2013-11-25 | Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013105203934 | 2013-10-29 | ||
| CN201310520393 | 2013-10-29 | ||
| CN201310520393.4 | 2013-10-29 | ||
| CN201310606778.2A CN103610639B (en) | 2013-10-29 | 2013-11-25 | Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103610639A CN103610639A (en) | 2014-03-05 |
| CN103610639B true CN103610639B (en) | 2016-03-09 |
Family
ID=50161374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310606778.2A Active CN103610639B (en) | 2013-10-29 | 2013-11-25 | Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103610639B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103990135B (en) * | 2014-05-13 | 2017-02-01 | 广东众生药业股份有限公司 | Micelle composed of taxane drug-loaded di-block polymer, and preparation method and application thereof |
| CN105982868B (en) * | 2015-12-23 | 2018-07-03 | 山东华铂凯盛生物科技有限公司 | A kind of La Luotasai water soluble powders injection and its application |
| CN105963430B (en) * | 2016-02-02 | 2019-10-22 | 侯君 | Chinese medicine composition and its preparation method and application |
| CN108434506B (en) * | 2018-04-27 | 2021-06-25 | 温州医科大学附属第二医院（温州医科大学附属育英儿童医院） | Expanded sponge loaded with nano sustained-release drug micelles, preparation method and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101972480A (en) * | 2010-01-19 | 2011-02-16 | 南京泛太化工医药研究所 | Docetaxel polymeric micelle medicine composition taking amino acid as stabilizing agent |
| CN102030871A (en) * | 2009-09-25 | 2011-04-27 | 天津大学 | Polyethylene glycol block biodegradable polyester comb-type graft copolymer, and preparation method and application thereof |
| CN102961322A (en) * | 2012-10-12 | 2013-03-13 | 杭州普施康生物科技有限公司 | Drug-loaded mixed micelle |
| CN103006539A (en) * | 2012-12-14 | 2013-04-03 | 沈阳药科大学 | Polymeric micelle medicine composition and preparation method thereof |
-
2013
- 2013-11-25 CN CN201310606778.2A patent/CN103610639B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102030871A (en) * | 2009-09-25 | 2011-04-27 | 天津大学 | Polyethylene glycol block biodegradable polyester comb-type graft copolymer, and preparation method and application thereof |
| CN101972480A (en) * | 2010-01-19 | 2011-02-16 | 南京泛太化工医药研究所 | Docetaxel polymeric micelle medicine composition taking amino acid as stabilizing agent |
| CN102961322A (en) * | 2012-10-12 | 2013-03-13 | 杭州普施康生物科技有限公司 | Drug-loaded mixed micelle |
| CN103006539A (en) * | 2012-12-14 | 2013-04-03 | 沈阳药科大学 | Polymeric micelle medicine composition and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103610639A (en) | 2014-03-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Pharmacokinetics, biodistribution, efficacy and safety of N-octyl-O-sulfate chitosan micelles loaded with paclitaxel | |
| CN101804021B (en) | Preparation method of polyene-containing taxol nanoparticle mixed micelle preparation and freeze-drying agent | |
| CN103772686A (en) | Amphiphilic block copolymer and preparation method thereof, micelle drug delivery system formed by copolymer and anti-tumor drug | |
| CN103006539B (en) | A kind of polymeric micelle medicine composition and preparation method thereof | |
| CN103610639B (en) | Containing the polymer micelle medicine carrying composition and method of making the same of buffer salt | |
| CN102357075A (en) | Docetaxel nano preparation and preparation method thereof | |
| CN103720675A (en) | Curcumin prodrug micelle with oxidation and reduction sensitivity, micellar monomer and preparation method of micellar monomer | |
| CN104856950A (en) | Paclitaxel micelle drug load system and preparation method thereof | |
| CN104856949A (en) | Docetaxel micelle drug load system and preparation method thereof | |
| CN102198083A (en) | Micelles and freeze-dried preparations of diblock polymer-supported taxane medicines and preparation and use thereof | |
| CN112156066B (en) | Preparation method of injectable composite hydrogel double-drug-loading system containing micelle | |
| CN103446040A (en) | Docetaxel-coated polymeric micelle and preparation method thereof | |
| CN102641245B (en) | Chitosan-chitosan derivative nanosphere for loading indissoluble medicament, preparation method of nanosphere, and application of nanosphere serving as oral prepration | |
| CN102670525A (en) | Application of ursolic-acid nano medicine-carrying microspheres for treating tumors and preparation | |
| CN102961322A (en) | Drug-loaded mixed micelle | |
| CN105560179A (en) | MPEG-PTMC diblock copolymer drug loaded micelle, preparation method and applications thereof | |
| CN103263672A (en) | Preparation method and application of nanometer particles of taxane drugs | |
| CN102274185A (en) | Antitumor pH-sensitive liposome and freeze-dried powder injection thereof, and preparation methods thereof | |
| CN107854430B (en) | Oral taxol nanosuspension and preparation method thereof | |
| CN103690512B (en) | A kind of deoxypodophyllotoxin polymer micelle lyophilized formulations | |
| CN102895669B (en) | Cis-platinum complex and preparation method thereof | |
| CN105310981A (en) | Docetaxel micelle drug loading system and preparation method and application thereof | |
| CN105288631A (en) | Novel anticancer drug nano-preparation and preparation method thereof | |
| CN103494829A (en) | Multifunctional nanoparticle preparation capable of preventing drug tolerance and preparation method thereof | |
| CN101693010B (en) | Cefathiamidine prosoma liposome preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |