CN102871966B - Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof - Google Patents
Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof Download PDFInfo
- Publication number
- CN102871966B CN102871966B CN2012104014343A CN201210401434A CN102871966B CN 102871966 B CN102871966 B CN 102871966B CN 2012104014343 A CN2012104014343 A CN 2012104014343A CN 201210401434 A CN201210401434 A CN 201210401434A CN 102871966 B CN102871966 B CN 102871966B
- Authority
- CN
- China
- Prior art keywords
- rapamycin
- acetone
- plga
- nano
- peg
- 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.)
- Expired - Fee Related
Links
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses nano drug carrier particles for improving the bioavailability of rapamycin and a preparation method thereof, so as to carry out a drug effect optimization. The nano drug carrier particles are prepared in the following steps that according to the principle of an emulsion solvent evaporation method, a determined amount of PEG-PLGA and rapamycin are respectively dissolved in acetone; after being mixed uniformly, the PEG-PLGA, the rapamycin and the acetone are slowly added into water to be stirred by magnetic force; after a certain period of time, the obtained liquid is ultrasonically homogenated, and organic phase is removed in a vacuum dryer; the obtained water phase removes free medicine in a centrifugal tube; and then an ultrafiltration tube is used for concentration, and nano particles are obtained after freezing and drying. The method has the advantages of convenience in operation, simplicity and feasibility, good repeatability and the like. The prepared nano drug carrier particles can improve the utilization rate of the medicine by improving the absorptivity of the medicine and prolonging the cycling time in a human body. Meanwhile, the nano particles which are prepared through the method have good biocompatibility, and surface active groups can further modify ligands or targeted groups.
Description
Technical field
The present invention relates to biomaterial and field of nanometer technology, be specifically related to a kind of have suitable envelop rate and drug loading, high biological safety, and can slowly discharge nanoscopic drug carriers of drug rapamycin and preparation method thereof.
Background technology
Rapamycin (Rapamycin) is the strong immunosuppressive drug with low nephrotoxicity, is widely used in transplant operation.Simultaneously it can effectively suppress again propagation and the transfer ability of vascular smooth muscle cell after blood vessel injury, thereby reduce the generation of vascular restenosis, and it can also promote myocardium damaged place cardiomyocyte proliferation, thereby very large application prospect is arranged aspect treating cardiovascular disease.But, due to the low-solubility of himself and the administering mode of present stage, greatly limited performance and the range of application of its drug effect.
Nano medicament carrying system has become the study hotspot in pharmaceutics and modern biomedical field as a kind of effective means of optimizing drug effect.Study more nano medicament carrying system and comprise nanometer liposome, nanosphere, nanocapsule, solid lipid nanoparticle and polymer capsule etc.At present, the study hotspot of nano medicament carrying system mainly concentrates on the exploitation for the diagnosis of cancer and targeted therapy preparation, and the part preparation has been arranged, and the stage enters clinical trial or comes into the market by experiment.cardiovascular disease is the same with cancer, human health in serious threat, but because part disease pathomechanism is explained imperfection, lack the reasons such as efficient targeting effect group, drug-loading system research for the cardiovascular disease diagnosis medicine lags far behind cancer, and many medicines are because poorly water-soluble, absorbance is low, bioavailability is low, the factors such as oral administration difficulty are not yet well applied, at present, the research of rapamycin nano-carrier still is in the starting stage both at home and abroad, domestic do not have a listing of effective rapamycin sustained-release oral preparation, in view of the good therapeutic effect of this medicine in many diseases field, accord with the demands of the market and clinical needs for a kind of suitable nano-carrier of this medicine exploitation.
This patent uses emulsion-solvent evaporation method to prepare the PEG-PLGA Nano microsphere and carries rapamycin, and similarly research has no report.PLGA and PEG are the high molecular polymer that possesses biological safety of FDA approval, and both synthetic block polymer vivo degradation product has no side effect to body.nanoparticle with the emulsion-solvent evaporation method preparation, internal layer is the core that is formed by physical action by hydrophobicity PLGA block and medicine, skin is hydrophilic PEG hydrated sheath, this hydrated sheath not only can solve the water solublity problem of insoluble drug rapamycin, can also help nano-particle to hide engulfing of reticuloendothelial system, reduce the adhesive attraction of serum albumin, thereby the time of staying of prolong drug in blood circulation, be conducive to the exploitation of oral slow-releasing preparation, avoided the high request of intravenous drug preparation Surfactant and dissolvent residual, potential biological safety risk and the inconvenience on administering mode.Because the PEG end can be active group, can further modify aglucon or other targeting group at nano grain surface by simple chemical reaction, reach the purpose of targeting pathological tissues, organ, thereby improve the drug level of lesions position, reduce the toxic and side effects of medicine to other histoorgan.In addition, PLGA is as a kind of biodegradable polymer, can slow degradation be the small-molecule substance of inanimate object toxicity in cyclic process, discharge simultaneously the medicine of parcel in degradation process, realize the slow release of medicine, can also control the rate of release of medicine by the ratio of lactic acid and hydroxyacetic acid in change PLGA, in the hope of meeting the administration requirements of different pharmaceutical, improve absorbance and the bioavailability of medicine, reach the maximization of drug effect.
Summary of the invention
Technical problem: the objective of the invention is by the nanometer synthetic technology, build the nano medicament carrying system for drug rapamycin, optimize existing drug administration mode, solve the problem of rapamycin water solublity, absorbability, targeting, bioavailability existence.The advantages such as another object of the present invention is to provide a kind of method for preparing above-mentioned rapamycin nano medicament carrying system, makes it have good reproducibility, and is easy to operate, simple, with low cost.
Technical scheme: in order to realize the foregoing invention purpose, the nanoparticle be used to improving the rapamycin bioavailability of the present invention is:
Use polyethylene glycol-polylactic acid hydroxyl ethanol acid copolymer PEG-PLGA as carrier, wherein the PEG end is methoxyl group, amino or carboxyl active group, form the internal package rapamycin with nucleocapsid structure by emulsion-solvent evaporation method, outside hydrophilic particle diameter is the nanoparticle of 100～120nm.
The preparation method of the nanoparticle be used to improving the rapamycin bioavailability of the present invention comprises the following steps:
1) take described copolymer p EG-PLGA and rapamycin, fully be dissolved in acetone respectively;
2) magnetic agitation with the acetone soln of described copolymer p EG-PLGA with after the acetone soln of rapamycin is mixed homogeneously, obtain organic liquid mixture, this organic liquid mixture is splashed in water under magnetic agitation, keep continuing to stir 30～60min, obtain the mixing material of organic facies and water;
3) with step 2) gained mixing material ultrasonic emulsification instrument supersound process 1～2min;
4) vacuum drying 3～6h removes acetone;
5) with step 4) remove acetone after remaining liq be transferred to the centrifugal free drug of removing in centrifuge tube;
6) shift supernatant to ultrafiltration and concentration nano-particle in super filter tube, residual acetone is removed in the ultra-pure water washing;
7) the gained concentrated solution uses the freezer dryer lyophilizing.
Wherein:
Step 1) in, copolymer p EG-PLGA and the rapamycin concentration ratio that feeds intake is 9-11: 1, and use acetone as solvent, the volume ratio of dissolving PEG-PLGA and rapamycin acetone used is 2: 1.
Step 2) in, the volume of water is 2: 1 with the ratio of organic liquid mixture volume, and the speed that organic facies splashes into water is 0.5mL/min.
Beneficial effect: the invention has the advantages that: adopt simple emulsion-solvent evaporation method to prepare nanoparticle, the method is with low cost, do not need super clean experimental situation, favorable repeatability, and can prepare in enormous quantities, have the advantages such as easy row convenient and simple for operation, with low cost and favorable repeatability, these advantages can be widely used in the nanoparticle preparation of various hydrophobic medicine.The nanoparticle good water solubility of preparation, have good biocompatibility and biological safety, and can be active group effectively prolong drug release time, and PEG end, is conducive to realize future target administration and prepares diagnosis and treatment one body preparation.
The specific embodiment
The present invention is described further below in conjunction with specific embodiment.
The preparation of rapamycin medicament-carried nano granule (RAPA-NPs): precision takes 100mg PEG-PLGA and 5mg rapamycin, fully is dissolved in respectively in 4mL and 2mL acetone.Magnetic agitation with the organic facies mix homogeneously after, with the speed of 0.5mL/min, the organic facies mixed liquor is splashed into 12mL aqueous phase under magnetic agitation, keep middling speed to continue to stir 30min.Gained liquid is removed organic facies with the dry 3h of the ultrasonic 1min of ultrasonic emulsification instrument (3s opens for instrument parameters: Φ 3 gear levers, 20%, 20 ℃ of power, and 1s closes) final vacuum.Liquid rotating is moved to centrifugal free drug (parameter: 3000rpm, 25 ℃, the 15min of removing in centrifuge tube, * 2), shift supernatant to ultrafiltration and concentration nano-particle (parameter: 4000rpm, 35 ℃ in super filter tube, 15min, * 2), residual organic solvent is removed in the ultra-pure water washing.Concentrated solution remain in 4 ℃ standby, get part and add 5% (w/v) sucrose as the freeze drying protectant mix homogeneously, lyophilizing in freezer dryer, weighing quality.
Nanoparticle size characterizes: use Particle Size Analyzer and dynamic light scattering to measure particle size and dispersibility, acquired results coincide, and the mean diameter that records is 112.25 ± 5.56nm, and the coefficient of dispersion is 0.248.
Vitro drug release: configure certain density RAPA-NPs solution 2mL, solvent is PBS: ethanol=9: 1, and put into 37 ℃ of calorstats and hatch, respectively at 4h, 7h, 20h, 30h, 42h, 52h, 70h takes out centrifugal (10000rpm, 25 ℃, 30min), draw supernatant 1.5mL and be stored in 4 ℃ of concentration to be measured, add the 1.5mL solvent and continue to hatch in nanoparticle precipitate.Use ultraviolet spectrophotometer to measure rapamycin in the supernatant of hatching the different time acquisition and, at the light absorption value at 277nm place, by the linear fit Equation for Calculating, go out concentration, draw the drug release curve, and carry out match, determine drug release model and equation thereof.Experimental result shows that within 20h, release amount of medicine arrives 40%, and slow release duration＞90h, show and can play good medicament slow release effect afterwards, and pharmaceutical release time-concentration relationship meets Higuchi and discharge model, meets the experiment expected results.
The cytotoxicity test: use CCK-8 test kit and schwann cell to carry out the cytotoxicity test, administration concentration is respectively 0.002,0.02,0.2,2,20 μ g/mL, and the continuous culture observation of cell is active.Test result shows, the rapamycin nanoparticle can suppress cell-proliferation activity to a certain extent, and inhibition strengthens with concentration in certain concentration range, but do not observe the reduction of cell quantity, show that this nano-particle does not have cell killing toxicity, has good biological safety.
Claims (1)
1. preparation method that is used for improving the nanoparticle of rapamycin bioavailability, it is characterized in that using polyethylene glycol-polylactic acid hydroxyl ethanol acid copolymer PEG-PLGA as carrier be used to the nanoparticle that improves the rapamycin bioavailability, wherein the PEG end is methoxyl group, amino or carboxyl active group, form the internal package rapamycin with nucleocapsid structure by emulsion-solvent evaporation method, outside hydrophilic particle diameter is the nanoparticle of 100～120nm, and the method comprises the following steps:
1) take described copolymer p EG-PLGA and rapamycin, fully be dissolved in acetone respectively;
2) magnetic agitation with the acetone soln of described copolymer p EG-PLGA with after the acetone soln of rapamycin is mixed homogeneously, obtain organic liquid mixture, this organic liquid mixture is splashed in water under magnetic agitation, keep continuing to stir 30～60min, obtain the mixing material of organic facies and water;
3) with step 2) gained mixing material ultrasonic emulsification instrument supersound process 1～2min;
4) vacuum drying 3～6h removes acetone;
5) step 4) is removed acetone after remaining liq be transferred to the centrifugal free drug of removing in centrifuge tube;
6) shift supernatant to ultrafiltration and concentration nano-particle in super filter tube, residual acetone is removed in the ultra-pure water washing;
7) the gained concentrated solution uses the freezer dryer lyophilizing;
In step 1), copolymer p EG-PLGA and the rapamycin concentration ratio that feeds intake is 9-11:1, uses acetone as solvent, and the volume ratio of dissolving PEG-PLGA and rapamycin acetone used is 2:1;
Step 2) in, the volume of water is 2:1 with the ratio of organic liquid mixture volume, and the speed that organic facies splashes into water is 0.5mL/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012104014343A CN102871966B (en) | 2012-10-19 | 2012-10-19 | Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012104014343A CN102871966B (en) | 2012-10-19 | 2012-10-19 | Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102871966A CN102871966A (en) | 2013-01-16 |
| CN102871966B true CN102871966B (en) | 2013-11-20 |
Family
ID=47473643
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012104014343A Expired - Fee Related CN102871966B (en) | 2012-10-19 | 2012-10-19 | Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102871966B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11779641B2 (en) | 2011-04-29 | 2023-10-10 | Selecta Biosciences, Inc. | Tolerogenic synthetic nanocarriers for allergy therapy |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EA201592106A3 (en) | 2013-05-03 | 2016-08-31 | Селекта Байосайенсиз, Инк. | LOCAL ACCOMPANYING INTRODUCTION OF TOLEROGENOUS SYNTHETIC NANOSATORS TO REDUCE HYPERSENSITIVITY TYPE I AND HYPERSENSITIVITY TYPE IV |
| WO2016037163A1 (en) | 2014-09-07 | 2016-03-10 | Selecta Biosciences, Inc. | Methods and compositions for attenuating gene therapy anti-viral transfer vector immune responses |
| PL3215133T3 (en) * | 2014-11-05 | 2021-06-14 | Selecta Biosciences, Inc. | Methods and compositions related to the use of low hlb surfactants in the production of synthetic nanocarriers comprising a rapalog |
| AU2018205496A1 (en) * | 2017-01-07 | 2019-07-25 | Selecta Biosciences, Inc. | Patterned dosing of immunosuppressants coupled to synthetic nanocarriers |
| BR112019018748A2 (en) | 2017-03-11 | 2020-04-07 | Selecta Biosciences Inc | methods and compositions related to combined treatment with anti-inflammatories and synthetic nanocarriers comprising an immunosuppressant |
| CN107049963B (en) * | 2017-06-23 | 2020-11-03 | 北京化工大学 | Sirolimus nano-drug composition and preparation method thereof |
| WO2019209883A1 (en) * | 2018-04-23 | 2019-10-31 | Graybug Vision, Inc. | Improved continuous microparticle manufacture |
| CN110623925B (en) * | 2019-09-26 | 2021-06-25 | 严鹏科 | Rapamycin nanometer sustained release agent and preparation method thereof |
| CN110693883A (en) * | 2019-11-21 | 2020-01-17 | 安徽医科大学第一附属医院 | Preparation method and application of rapamycin-entrapped zinc-organic framework drug delivery system |
| CN114767659A (en) * | 2022-04-27 | 2022-07-22 | 中山大学孙逸仙纪念医院 | Medicinal preparation targeting PITPNM3 and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1771910A (en) * | 2005-11-02 | 2006-05-17 | 浙江大学 | Nanometer particle of insoluble medicine and its prepn |
| CN102266291A (en) * | 2011-06-30 | 2011-12-07 | 上海中医药大学附属普陀医院 | Preparation method of strychnine immune nanoparticles |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2012501965A (en) * | 2008-06-16 | 2012-01-26 | バインド バイオサイエンシズ インコーポレイテッド | Drug-loaded polymer nanoparticles and methods for producing and using the same |
-
2012
- 2012-10-19 CN CN2012104014343A patent/CN102871966B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1771910A (en) * | 2005-11-02 | 2006-05-17 | 浙江大学 | Nanometer particle of insoluble medicine and its prepn |
| CN102266291A (en) * | 2011-06-30 | 2011-12-07 | 上海中医药大学附属普陀医院 | Preparation method of strychnine immune nanoparticles |
Non-Patent Citations (1)
| Title |
|---|
| 苗立夫，等.雷帕霉素聚乳酸-聚乙醇酸纳米粒子的制备、表征及血管内局部给药效能.《中国医学科学院学报》.2008,第30卷(第4期),第491-497页. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11779641B2 (en) | 2011-04-29 | 2023-10-10 | Selecta Biosciences, Inc. | Tolerogenic synthetic nanocarriers for allergy therapy |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102871966A (en) | 2013-01-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102871966B (en) | Nano drug carrier particles for improving bioavailability of rapamycin and preparation method thereof | |
| Yao et al. | Loading-free supramolecular organic framework drug delivery systems (sof-DDSs) for doxorubicin: normal plasm and multidrug resistant cancer cell-adaptive delivery and release | |
| Dhanka et al. | Injectable methotrexate loaded polycaprolactone microspheres: physicochemical characterization, biocompatibility, and hemocompatibility evaluation | |
| CN110215438B (en) | Preparation method and application of mesoporous silicon nanoparticles carrying anthracyclines and photosensitizers | |
| CN104189916A (en) | Multimer alhumin nanospheres, and preparation method and application thereof | |
| CN111135296B (en) | Albumin-bound indocyanine green anti-tumor photo-thermal preparation and preparation method thereof | |
| Yang et al. | NIR-activated self-sensitized polymeric micelles for enhanced cancer chemo-photothermal therapy | |
| Chen et al. | Glucosamine derivative modified nanostructured lipid carriers for targeted tumor delivery | |
| CN112870191B (en) | Preparation method of metal organic frame ZIF-8 wrapped Prussian blue quercetin-loaded nanoparticles | |
| CN107412787B (en) | Photosensitizer targeted nanoparticle for optical treatment and preparation method and application thereof | |
| Gao et al. | AuNRs@ MIL-101-based stimuli-responsive nanoplatform with supramolecular gates for image-guided chemo-photothermal therapy | |
| CN104940945A (en) | Hollow mesoporous copper sulfide compound modified by hyaluronic acid and preparation method and application thereof | |
| CN101984958B (en) | Nanoscale albendazole micropowder and preparation method thereof | |
| Shi et al. | Copper (II)-disulfiram loaded melanin-dots for cancer theranostics | |
| CN104288111A (en) | Ginsenoside Rg3 poly(lactic-co-glycolic acid) nano microsphere and preparation method thereof | |
| Chen et al. | Doxorubicin-encapsulated thermosensitive liposome-functionalized photothermal composite scaffolds for synergistic photothermal therapy and chemotherapy | |
| CN107126561B (en) | Anti-tumor synergistic composition capable of realizing combined treatment of chemotherapy and PTT/PDT and application thereof | |
| CN108912349A (en) | Polylactic acid microsphere and preparation method thereof and the application in medicament slow release | |
| CN111632032A (en) | Natural small molecule co-assembled nano-drug delivery system and preparation method and application thereof | |
| CN109078184B (en) | Loaded double-drug nanoparticle and preparation method and application thereof | |
| Ostovar et al. | Nanocomposite of chitosan/gelatin/carbon quantum dots as a biocompatible and efficient nanocarrier for improving the Curcumin delivery restrictions to treat brain cancer | |
| Kamboj et al. | Preparation and characterization of metformin loaded Stearic Acid coupled F127 Nanoparticles | |
| CN112386585B (en) | Self-assembled nano-drug and preparation method and application thereof | |
| CN104434792A (en) | Polymer micelle, preparation method thereof, antitumor pharmaceutical composition, preparation and preparation method thereof | |
| WO2023193389A1 (en) | Resveratrol-lecithin nanoparticle, method for preparing same, and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | 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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131120 Termination date: 20161019 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |