CN103099784B - Nanometer medicine particle, preparation method and application thereof - Google Patents
Nanometer medicine particle, preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a nanometer medicine particle. The nanometer medicine particle comprises a hydrophobic polymer, a single-layer lipid molecule, an amphipathy macromolecular compound, at least one chemotherapy drug attached to the hydrophobic polymer, and at least one near-infrared photothermal conversion agent, wherein the hydrophobic polymer, the at least one chemotherapy drug attached to the hydrophobic polymer, and the at least one near-infrared photothermal conversion agent form a hydrophobic core; the single-layer lipid molecule surrounds the surface of the hydrophobic core to form an intermediate layer; and the amphipathy macromolecular compound intersperses in the intermediate layer to form a shell. The invention also provides a preparation method of the nanometer medicine particle. The nanometer medicine particle can realize the combined treatment of thermotherapy and chemotherapy and has high biocompatibility; and the preparation method is simple, convenient and easy to carry out.
Description
Technical field
The present invention relates to pharmaceutical carrier field, relate in particular to a kind of Nano medication granule and preparation method thereof.
Background technology
Organize normally homeostasis to realize by the heterogeneous equilibrium between cell proliferation rate and cell death.This balance, because the increase of cell proliferation rate or the reduction of cell mortality destroy, causes the misgrowth of cell, is the main matter that causes formation of cancer.Routine strategy in treatment of cancer comprises chemotherapy, radiotherapy, operation, biotherapy or their combination.
Chemotherapy, utilizes chemicals to kill the propagation of tumor cell, inhibition tumor cell and the differentiation of promotion tumor cell, is a kind of measure of important treatment cancer.For example: amycin (Doxorubicin) is a kind of antitumor antibiotics, can suppress the synthetic of RNA and DNA, the tumor cell of various growth cycles is all had to inhibitory action, be widely used in the chemotherapy of various tumors.
Near-infrared tumor thermotherapy mainly utilizes near infrared light thermal agent to absorb near infrared light, improves the local temperature of whole body or tumor tissues, utilizes heat effect and secondary effect thereof to kill tumor cell, reaches the object for the treatment of tumor.Near infrared light thermal agent as: indocyanine green (Indocyanine Green, ICG) be a kind of near infrared light thermal agent that can be used for clinical treatment diagnosis of being ratified by U.S. food Drug Administration, it has characteristic absorption peak in near-infrared region.In the treatment of cancer, often adopt conjoint therapy, as the use in conjunction of radiation and chemotherapy.The high heat of near-infrared thermotherapy can make tumor cell membrane be damaged, the permeability changes of tumor cell cell membrane, and chemotherapeutics more easily enters tumor cell, and tumor cell chemotherapy medicine intake increases, and drug level increases; High heat has also suppressed the synthetic of DNA of tumor cell, RNA and protein simultaneously, has increased the effect of the chemotherapeutics such as amycin; High heat also can suppress the reparation of the cell injury that chemotherapeutics causes.In view of above-mentioned, develop the conjoint therapy of near-infrared thermotherapy and chemotherapy, the common transmission that realizes chemotherapeutics and near-infrared thermotherapy photo-thermal reagent becomes problem demanding prompt solution.Nanotechnology provides new opportunity for common transmission chemotherapeutics and near-infrared thermotherapy photo-thermal reagent.Polymer nano granules and nanometer liposome are that two class main flow nano-carriers of representative can efficiently wrap up and transmit medicine, gene etc., become various countries scientist's study hotspot.The polymer nano granules such as Poly(D,L-lactide-co-glycolide (PLGA), polylactic acid, poly-second lactone have good biocompatibility, biodegradation is controlled and catabolite toxicity is low, easily realize targeting controls the advantages such as release, nanometer liposome has the biofilm structure of phospholipid bilayer, there is fabulous hydrophilic, lipotropy and natural targeting, long-lasting, pardon, and nontoxic, non-immunogenicity, infiltration rate is fast, bioavailability is high, be easy to the advantages such as surface-functionalized.Prior art CN200910188814.1 discloses a kind of nano-particle and preparation method thereof, the intermediate layer that forms kernel, phospholipid formation with Poly(D,L-lactide-co-glycolide (PLGA) is around core surface, and the housing parts that contains the PEG-DSPE formation of amino or carboxyl is interspersed in described intermediate layer.Prior art CN201010607708.5 discloses a kind of fluorescent nano probe, also be to form intermediate layer that kernel, phospholipid form around described core surface with Poly(D,L-lactide-co-glycolide, the housing parts that contains PEG-DSPE (DSPE-PEG) formation of amino or carboxyl is interspersed in described intermediate layer, in core, be dispersed with indocyanine green within it, this fluorescent probe can be identified tumor cell, for the early diagnosis of tumor.What in above two kinds of prior aries, the preparation method of nano-particle all adopted is the mode being uniformly mixed, and need to reach the response time of a few hours, and preparation speed is slower.
Summary of the invention
The present invention is intended to solve above-mentioned problems of the prior art, a kind of Nano medication granule is proposed, comprise hydrophobicity polymer, monolayer lipid molecule, amphiphilic macromolecular compound and change reagent with at least one chemotherapeutics and at least one the near-infrared photo-thermal of the absorption of hydrophobicity polymer, wherein, described hydrophobicity polymer forms hydrophobic cores with at least one chemotherapeutics and at least one the near-infrared photo-thermal conversion reagent that are adsorbed on hydrophobicity polymer; Monolayer lipid molecule is looped around described hydrophobic cores surface and forms intermediate layer, and described monolayer lipid molecule has towards the hydrophobic part of described hydrophobic cores with towards the hydrophilic segment of described Nano medication granule outside; Described amphiphilic macromolecular compound is interspersed in described intermediate layer and forms hydrophilic shell, described amphiphilic macromolecular compound be have fat end and with the compound of the water-wet side of described fat end covalent cross-linking, described fat end can help described amphiphilic macromolecular compound to insert described monolayer lipid molecule layer, and described water-wet side extends in described Nano medication granule outside.
Preferably, described hydrophobicity polymer is selected from Poly(D,L-lactide-co-glycolide, polylactic acid, polycaprolactone.
Preferably, described monolayer lipid molecule is selected from lecithin, cephalin.
Preferably, described amphiphilic macromolecular compound is PEG-DSPE-maleic amide.
Preferably, described chemotherapeutics is selected from amycin, epirubicin, paclitaxel, NVB, cisplatin.
Preferably, described near-infrared photo-thermal conversion reagent is selected from indocyanine green, gold nanorods, CNT.
Preferably, to change the mass ratio of reagent be 1: 1 for described chemotherapeutics and described near-infrared photo-thermal.
Preferably, described chemotherapeutics and described near-infrared photo-thermal are changed the 25%-75% that the quality sum of reagent is described hydrophobic polymer quality.
The present invention also provides a kind of method of preparing Nano medication granule, it is characterized in that, comprises the following steps:
(1) hydrophobicity polymer is dissolved in second cyanogen, obtains hydrophobicity polymer second cyanogen solution;
(2) monolayer lipid molecule, amphiphilic macromolecular compound, chemotherapeutics, near-infrared photo-thermal conversion reagent are dissolved in ethanol water, obtain mixed ethanol aqueous solution;
(3) hydrophobicity polymer second cyanogen solution is dropwise joined in mixed ethanol aqueous solution and reacted, with ultrasound wave, carry out ultrasonicly, can obtain Nano medication granule.
The preparation method of Nano medication granule of the present invention also comprises twice of super filter tube centrifugal ultrafiltration of the solution after ultrasonic.
Further aspect of the present invention provides the application in preparing antitumor drug of described Nano medication granule.
Beneficial effect of the present invention is, the first, and Nano medication granule of the present invention can be delivered to tumor locus by chemotherapeutics and near-infrared photo-thermal conversion reagent simultaneously, realizes the therapeutic alliance of thermotherapy and chemotherapy; The second, the hydrophobicity polymer comprising in Nano medication granule of the present invention, monolayer lipid molecule and amphiphilic macromolecular compound all have good biocompatibility, can carry out biodegradation and are absorbed or got rid of external by normal physiological pathway; The 3rd, Nano medication granule has significantly improved the light stability of near-infrared photo-thermal conversion reagent; The 4th, the preparation method of Nano medication granule of the present invention is simple and easy to do, convenient operation; The 5th, the solution of the present invention combines nano-encapsulated technology, chemotherapeutics and for the near-infrared photo-thermal of thermotherapy, changes reagent three's advantage, the effectively growth of inhibition tumor cell.
Accompanying drawing explanation
Fig. 1 is the structural representation of the Nano medication granule prepared in the embodiment of the present invention.
Fig. 2 is the electron-microscope scanning figure of the Nano medication granule prepared in the embodiment of the present invention.
Fig. 3 is the results of animal figure of the Nano medication granule therapy effect prepared in the embodiment of the present invention.
The specific embodiment
In order to make those skilled in the art better understand the application's technical scheme, below in conjunction with the accompanying drawing in the embodiment of the present application, the technical scheme in the embodiment of the present application is carried out to clear, complete description.
The solution of the present invention combines nano-encapsulated technology, chemotherapeutics and for the near-infrared photo-thermal of thermotherapy, changes reagent three's advantage, the effectively growth of inhibition tumor cell.EPR (the enhanced permeability and retention effect) effect of utilizing Nano medication to send, improves the targeting of medicine to tumor cell, reduces cytotoxicity, and the action time of prolong drug and effect; By the near-infrared photo-thermal conversion reagent for thermotherapy and the combination of chemotherapeutics, produce cooperative effect, thus the more effectively growth of inhibition tumor cell, and kill tumor cell.
The kernel of Nano medication granule of the present invention has hydrophobicity, hydrophobicity polymer, consists of, and described hydrophobicity polymer can be adsorbed in its surface by chemotherapeutics and near-infrared photo-thermal conversion reagent simultaneously.For example, hydrophobicity polymer of the present invention can be selected Poly(D,L-lactide-co-glycolide (PLGA), polylactic acid (PLA), polycaprolactone etc., but does not get rid of other available hydrophobic polymers.The medicine containing in kernel can be the combination of at least one chemotherapeutics and at least one near-infrared photo-thermal conversion reagent, but do not get rid of and also comprise other drug, as genomic medicine or antitumor adjuvant therapy medicaments such as siRNA, miRNA, antioncogenes, as ondansetron, calcium folinate etc.
Conventional chemotherapeutics comprises antitumor antibiotics, as amycin, epirubicin; Antitumor animals and plants component drugs class, as paclitaxel, NVB etc.; Antimetabolite, as 5-fluorouracil etc.; Alkylating agent, as cyclophosphamide etc.; Antitumor hormones, as atamestane etc.; Miscellany, as cisplatin etc.
Conventional near-infrared photo-thermal conversion reagent mainly comprises indocyanine green, gold nanorods, CNT, can absorb near infrared light, carry out photo-thermal conversion, tumor cell temperature is raise, because can making tumor cell membrane, Gao Re is damaged, the permeability changes of tumor cell cell membrane, and chemotherapeutics more easily enters tumor cell, tumor cell chemotherapy medicine intake increases, and drug level increases; High heat has also suppressed the synthetic of DNA of tumor cell, RNA and protein simultaneously, has increased the effect of chemotherapeutics; High heat also can suppress the reparation of the cell injury that chemotherapeutics causes, therefore, near-infrared photo-thermal conversion reagent and chemotherapeutics exist the synergy that can bring into play thermotherapy and chemotherapy simultaneously, greatly improve the lethality to tumor cell.
Monolayer lipid molecule can be selected lecithin, cephalin.
Nano medication granule shell consists of amphiphilic macromolecular compound, for improving the water stability of Nano medication granule, protection Nano medication granule action time in vivo, and can chemical modification so that the functions such as targeting of Nano medication to be provided.The water-wet side target tumor cell of amphiphilic macromolecular compound, its fat end can help molecule to insert monolayer lipid molecule layer, water-wet side and fat end covalent cross-linking, target tumor cell.For example, amphiphilic macromolecular can be PEG-DSPE-maleic amide (DSPE-PEG-MAL).The fat end of amphiphilic macromolecular compound is divided with water-wet side part and can further be modified, as modified with folic acid or ligand molecular that can specific binding tumor surface receptor, to improve the targeting to tumor cell.
As shown in Figure 1, Nano medication granule in present embodiment, for wrapping altogether the Nano medication granule that carries indocyanine green and amycin, comprises amycin, indocyanine green (ICG), Poly(D,L-lactide-co-glycolide (PLGA), phospholipid, PEG-DSPE-maleic amide (DSPE-PEG-MAL).Wherein, Poly(D,L-lactide-co-glycolide (PLGA) is as kernel parcel amycin and indocyanine green (ICG), phospholipid is looped around core surface and forms intermediate layer, and PEG-DSPE-maleic amide (DSPE-PEG-MAL) part is interspersed in intermediate layer and forms shell.PLGA has hydrophobicity, as kernel, amycin and indocyanine green is wrapped in wherein, can effectively strengthen the stability of indocyanine green, can control amycin and indocyanine green simultaneously in the release of tumor locus.Phospholipid is surrounded on PLGA surface and forms intermediate layer, the hydrophobic layer of phospholipid and the effect of PLGA core surface, the outer surface in intermediate layer is hydrophilic single layer structure, can avoid immune identification, DSPE-PEG-MAL is interspersed in phospholipid and forms shell, makes whole Nano medication granule have stability.
The preparation method of wrapping altogether the Nano medication granule that carries indocyanine green and amycin in present embodiment is as follows:
(1) Poly(D,L-lactide-co-glycolide is dissolved in second cyanogen, obtains Poly(D,L-lactide-co-glycolide second cyanogen solution;
(2) phospholipid, DSPE-PEG-MAL, amycin, indocyanine green are dissolved in ethanol water, obtain mixed ethanol aqueous solution;
(3) Poly(D,L-lactide-co-glycolide second cyanogen solution is dropwise joined in mixed ethanol aqueous solution and reacted, with ultrasound wave, carry out ultrasonicly, can obtain the Nano medication granule that common bag carries indocyanine green and amycin.
The experimental technique using in following embodiment if no special instructions, is conventional method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
embodiment 1
(1) Poly(D,L-lactide-co-glycolide is dissolved in second cyanogen, obtains Poly(D,L-lactide-co-glycolide second cyanogen solution, and concentration is 2mg/mL;
(2) 180 μ g phospholipid, 120 μ gDSPE-PEG-MAL, 750 μ g amycin, 750 μ g indocyanine green are dissolved in 3mL4% ethanol water, obtain mixed ethanol aqueous solution, and insoluble sinkage appears in the indocyanine green of its Green;
(3) 1mL Poly(D,L-lactide-co-glycolide second cyanogen solution is dropwise joined in 3mL mixed ethanol aqueous solution and reacted, adopt ultrasonic cell disruption instrument to carry out ultrasonic with the frequency of 20KHz and the power of 130W, ultrasonic time is 5min, after ultrasonic, precipitation in step (2) disappears, and solution is dirty-green;
(4) the super filter tube centrifugal ultrafiltration twice with 10KDa by the solution after ultrasonic, can obtain the Nano medication granule that common bag carries indocyanine green and amycin.
The Nano medication granule that the common bag obtaining carries indocyanine green and amycin as shown in Figure 2, can find out, prepared Nano medication granule is spherical in shape, and dispersibility is better.The mean diameter that records Nano medication granule with Particle Size Analyzer is 80-120nm.Utilize following formula to calculate respectively the envelop rate of amycin and indocyanine green: EN%=(1-Cf/Ct) * 100%, wherein, Cf is the amount of free drug, Ct is the total amount of medicine, obtaining prepared Nano medication granule is 35.75 ± 1.37% to the envelop rate of amycin, to the envelop rate of indocyanine green, is 32.73 ± 1.17%.
comparative example 1
(1) Poly(D,L-lactide-co-glycolide is dissolved in second cyanogen, obtains Poly(D,L-lactide-co-glycolide second cyanogen solution, and concentration is 2mg/mL;
(2) 180 μ g phospholipid, 120 μ gDSPE-PEG-MAL, 750 μ g indocyanine green are dissolved in 3mL4% ethanol water, obtain mixed ethanol aqueous solution, and insoluble sinkage appears in the indocyanine green of its Green;
(3) 1mL Poly(D,L-lactide-co-glycolide second cyanogen solution is dropwise joined in 3mL mixed ethanol aqueous solution and reacted, adopt ultrasonic cell disruption instrument to carry out ultrasonic with the frequency of 20KHz and the power of 130W, ultrasonic time is 5min, after ultrasonic, precipitation in step (2) disappears, and solution is dirty-green;
(4) the super filter tube centrifugal ultrafiltration twice with 10KDa by the solution after ultrasonic, can obtain the Nano medication granule that bag carries indocyanine green.
bag carries the zoopery of the Nano medication granule therapy effect of indocyanine green and amycin altogether
Laboratory animal adopts 4-6 week, and the female BALB/c nude mice that body weight is 15-20g, to mouse bare subcutaneous injection 1 * 10
6individual MCF-7 breast cancer cell, treats that gross tumor volume reaches 300mm
3during left and right, use, gross tumor volume computing formula is: length of tumor * (tumor width)
2/ 2.Be 90 days experimental period.
Experiment grouping:
A, blank group
At tumor locus, inject 150 μ L PBS solution; Use 1W/cm
2the 808nm near-infrared laser of power density irradiate 5min.
B, thermotherapy group
At tumor locus, inject the Nano medication particle solution that bag that 150 μ L are prepared according to the preparation method in comparative example 1 carries indocyanine green, wherein, the concentration of indocyanine green is 55 μ g/mL; Use 1W/cm
2the 808nm near-infrared laser of power density irradiate 5min.
C, thermotherapy chemotherapy combined treatment group
At tumor locus, inject the Nano medication particle solution that common bag that 150 μ L are prepared according to the preparation method in embodiment 1 carries indocyanine green and amycin, wherein, the concentration of indocyanine green is 55 μ g/mL, and the concentration of amycin is 60 μ g/mL; Use 1W/cm
2the 808nm near-infrared laser of power density irradiate 5min.
Experimental result as shown in Figure 3, has only been injected nude mice tumor Fast Growth in 17 days of the blank group of PBS.At the 5th day, in tumor locus, there is black crust in the nude mice of thermotherapy group and therapeutic alliance group, subsequently, thermotherapy group nude mice in black crust, around occur at the 17th day, occurring enlargement by tumor growth; Therapeutic alliance group tumor after the 5th day starts to diminish, and to the 17th day, disappears, and continues to observe to the 90th day, has no tumor recurrence.Experimental result shows, wraps altogether the Nano medication granule of year indocyanine green and amycin inhibition mouse tumor is grown and had good effect.
Above-described embodiment of the present invention, does not form limiting the scope of the present invention.Any modification of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in claim protection domain of the present invention.
Claims (8)
1. a Nano medication granule, comprise hydrophobicity polymer, monolayer lipid molecule, amphiphilic macromolecular compound and change reagent with at least one chemotherapeutics and at least one the near-infrared photo-thermal of the absorption of hydrophobicity polymer, wherein, described hydrophobicity polymer forms hydrophobic cores with at least one chemotherapeutics and at least one the near-infrared photo-thermal conversion reagent that are adsorbed on hydrophobicity polymer, monolayer lipid molecule is looped around described hydrophobic cores surface and forms intermediate layer, and described amphiphilic macromolecular compound is interspersed in described intermediate layer and forms shell; The mass ratio that described chemotherapeutics and described near-infrared photo-thermal are changed reagent is 1:1; Described chemotherapeutics and described near-infrared photo-thermal are changed the 25%-75% that the quality sum of reagent is described hydrophobic polymer quality.
2. Nano medication granule according to claim 1, is characterized in that, described hydrophobicity polymer is selected from Poly(D,L-lactide-co-glycolide, polylactic acid, polycaprolactone.
3. Nano medication granule according to claim 1, is characterized in that, described monolayer lipid molecule is selected from lecithin, cephalin.
4. Nano medication granule according to claim 1, is characterized in that, described amphiphilic macromolecular compound is PEG-DSPE-maleic amide.
5. Nano medication granule according to claim 1, is characterized in that, described chemotherapeutics is selected from amycin, epirubicin, paclitaxel, NVB, cisplatin.
6. Nano medication granule according to claim 1, is characterized in that, described near-infrared photo-thermal conversion reagent is selected from indocyanine green, gold nanorods, CNT.
7. a method of preparing Nano medication granule as claimed in claim 1, is characterized in that, comprises the following steps:
(1) hydrophobicity polymer is dissolved in second cyanogen, obtains hydrophobicity polymer second cyanogen solution;
(2) monolayer lipid molecule, amphiphilic macromolecular compound, chemotherapeutics, near-infrared photo-thermal conversion reagent are dissolved in ethanol water, obtain mixed ethanol aqueous solution;
(3) hydrophobicity polymer second cyanogen solution is dropwise joined in mixed ethanol aqueous solution and reacted, with ultrasound wave, carry out ultrasonicly, can obtain Nano medication granule.
8. the preparation method of Nano medication granule according to claim 7, is characterized in that, also comprises twice of super filter tube centrifugal ultrafiltration of the solution after ultrasonic.
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Effective date of registration: 20201217 Address after: 519000 12-storey factory building of Zone A, No. 1 Gangle Road, Tangjiawan Town, Zhuhai High-tech Zone, Guangdong Province Patentee after: ZHUHAI INSTITUTE OF ADVANCED TECHNOLOGY CHINESE ACADEMY OF SCIENCES Co.,Ltd. Address before: 1068 No. 518055 Guangdong city in Shenzhen Province, Nanshan District City Xili University School Avenue Patentee before: SHENZHEN INSTITUTES OF ADVANCED TECHNOLOGY |