CN110665003A - Double-drug-loading carrier-free nanoparticle and preparation method thereof - Google Patents
Double-drug-loading carrier-free nanoparticle and preparation method thereof Download PDFInfo
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- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
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- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
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- A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
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- A61P35/00—Antineoplastic agents
Abstract
The invention discloses a double-drug-loading carrier-free nanoparticle preparation and a preparation method thereof, wherein the double-drug-loading carrier-free nanoparticle is not added with any carrier and only consists of a phototherapeutic drug and a chemotherapeutic drug, wherein the phototherapeutic drug is a drug with photosensitizer property: chlorin e 6; the chemotherapeutic drugs are camptothecin drugs: one of camptothecin, 10-hydroxycamptothecin or 7-ethyl-10-hydroxycamptothecin. The double-drug-loading carrier-free nanoparticle is prepared by combining an anti-solvent precipitation method with a high-pressure homogenization method or combining an alkali-soluble acid precipitation method with a high-pressure homogenization method, the particle size is 140-170 nm, a freeze-drying protective agent can be added for freeze-drying, and the particle size has no significant difference before and after freeze-drying and redissolving. The double-drug-loading carrier-free nanoparticle realizes the synergistic effect of phototherapy-chemotherapy, has obvious anti-tumor effect, avoids the problems of undefined safety and in-vivo metabolism and the like caused by carrier synthesis, has good biocompatibility and has potential clinical application value.
Description
Technical Field
The invention belongs to the technical field of medicines, particularly relates to a double-medicine-carrying carrier-free nanoparticle preparation, and particularly relates to a double-medicine-carrying carrier-free nanoparticle integrating photodynamic therapy and chemotherapy and a preparation method thereof.
Background
Malignant tumors are major diseases threatening human health and life, and can destroy the structure and function of human tissues and organs, causing necrotic hemorrhage with infection, and patients may eventually die due to exhaustion of organ functions. The current treatment means of tumors mainly comprise surgical operation, radiotherapy and chemotherapy. The surgical operation treatment mainly aims at the tumors which are not diffused, such as solid tumors, and has certain sequelae and dysfunction after the operation. The period of radiotherapy and chemotherapy is long, and the treatment has a certain limit, and the traditional Chinese medicine composition is easy to generate large toxic and side effects during treatment and has great damage to normal cells and tissues of a human body. In recent years, photodynamic therapy has been developed rapidly as a non-operative alternative means for clinical treatment of malignant tumors, which mainly utilizes photosensitizer accumulated in tumor site, and after being excited by light irradiation with specific wavelength, generates active oxygen molecules, free radicals and other active substances to cause apoptosis or necrosis of tumor cells, and destroys the capillary circulation system in tumor tissues to make tumor cells lack oxygen or lack nutrition and fail.
Chlorin e6 (chlorin e6, Ce 6) has strong photosensitization effect and low toxicity, and is one of the commonly used photosensitizers in photodynamic therapy. But the water solubility is poor, the bioavailability is low, the in-vivo distribution is not selective, and the clinical application is limited. Camptothecin (CPT) is a derivative of indole alkaloid separated from fruit of Camptotheca acuminata (Camptotheca acuminata Decaisne) of davidiaceae, which is a specific Chinese herb. It can kill tumor cells by inhibiting topoisomerase I, is a broad-spectrum antitumor drug, and has certain curative effects on non-small cell lung cancer, ascetic liver cancer, gastric cancer, breast cancer, pancreatic cancer, bladder cancer, leukemia, etc. However, camptothecin drugs have poor water solubility and low bioavailability, and the clinical application of the camptothecin drugs is limited. Under the alkaline condition, the lactone ring of the camptothecin medicament can be opened to form sodium salt, the water solubility is increased, and the camptothecin medicament is clinically prepared into sodium salt injection or powder injection by an alkalinization ring-opening method by utilizing the characteristic of the camptothecin medicament. However, the lactone ring opening increases the water solubility of the drug and simultaneously reduces the half-life of the drug, and more importantly, the active group of the drug is destroyed, so that the drug effect is remarkably reduced and the toxic and side effects are remarkably increased.
In order to overcome the defects, further improve the cure rate of tumor patients and make up for the defects of monotherapy, the combination application of the chemotherapy and the phototherapy by adopting the nano drug delivery system has more clinical significance. Nanosudministration systems are those in which the drug is formulated with a suitable carrier using appropriate techniques into nanometer-sized particles, typically of the order of 10-500 nm in size. Because of the huge specific surface area, the dissolution of the medicine can be improved, thereby increasing the absorption speed and the absorption rate of the medicine, improving the bioavailability of the medicine, protecting the medicine from being degraded by organisms and having influence on the pharmacokinetics and the tissue distribution behavior of the medicine in vivo. And the nanometer-level particle size is beneficial to passively targeting to tumor and inflammation parts through an EPR effect, and can prolong the in vivo circulation time so as to avoid phagocytosis by reticuloendothelial systems (RES systems) such as liver and spleen, so that the nanometer-level particle size is widely used for clinical administration of insoluble antitumor drugs and is popular with research institutions of various countries in the world since being reported. Through research and development for decades, different varieties are commercialized successively and are updated continuously, so that the varieties are better suitable for treatment of diseases.
Most of the existing nano drug delivery systems still cannot get rid of the dependence on the traditional synthetic carrier materials. This not only makes it unable to carry medicine with high efficiency, but also may bring the safety problem in its clinical application process. The double-drug-loading carrier-free nanoparticle is a pure drug nano colloidal dispersion system, does not need a carrier material, is assembled only by the interaction among drug molecules such as pi-pi conjugation, electrostatic attraction and hydrophobic interaction, and achieves a better tumor inhibition effect by the EPR effect of small-particle-size particles and the synergistic effect of phototherapy-chemotherapy combination therapy.
Disclosure of Invention
The invention mainly aims to provide double-drug-loading carrier-free nanoparticles, which greatly improve the anti-tumor effect through the synergistic effect of a photosensitizer and chemotherapeutic drug double-drug molecules.
The technical scheme adopted for realizing the purpose of the invention is as follows:
the invention provides a double-drug-loading carrier-free nanoparticle which is formed by assembling drug molecules between phototherapeutic drugs and chemotherapeutic drugs without adding any carrier, wherein the interaction between the drug molecules comprises pi-pi conjugation, electrostatic attraction and hydrophobic interaction.
Further, the phototherapeutic agent is a drug with photosensitizer property: chlorin e 6; the chemotherapeutic drugs are camptothecin drugs: one of camptothecin, 10-hydroxycamptothecin or 7-ethyl-10-hydroxycamptothecin.
Furthermore, the molar ratio of the photosensitizer drug molecules to the camptothecin drug molecules is 1:1, 1:2, 1:3 and 1: 4.
Further, a preparation method of the double-drug-loading carrier-free nanoparticle is realized by combining an anti-solvent precipitation method with a high-pressure homogenization method or combining an alkali-soluble acid precipitation method with a high-pressure homogenization method.
Further, the anti-solvent precipitation method is combined with a high-pressure homogenization method, and the method comprises the following specific steps: (1) co-dissolving photosensitizer and chemotherapeutic medicine in organic solvent to form organic phase; (2) the organic phase is added into the water phase drop by drop under the ultrasonic condition; (3) continuing to perform ultrasonic treatment for 10min at low temperature; (4) dialyzing to remove the organic solvent; (5) homogenizing under high pressure for several times.
Further, in the preparation process of the double-drug-loading carrier-free nanoparticle, the organic phase is one or two of DMF (dimethyl formamide) or DMSO (dimethyl sulfoxide), the water phase is deionized water or a glucose solution with the mass fraction of glucose being 5%, and the volume ratio of the organic phase to the water phase is 1:10 ~ 1: 15.
Further, the alkali-dissolution acid-precipitation method and the high-pressure homogenization method are combined, and the method comprises the following specific steps: (1) dissolving a certain amount of photosensitizer and chemotherapeutic drug in 10 mL of NaOH solution; (2) the solution was added dropwise to HCl solution under sonication to pH = 5; (3) continuing to perform ultrasonic treatment for 10min at low temperature; (4) centrifuging at 10000 rpm for 10min, discarding supernatant, and resuspending the precipitate in deionized water; (5) homogenizing under high pressure for several times.
Further, in the preparation process of the double-drug-loading carrier-free nanoparticle, the concentrations of NaOH solution and HCl solution are both 0.2M.
Furthermore, the ultrasonic power of the anti-solvent precipitation method combined with the high-pressure homogenization method and the alkali-soluble acid precipitation method combined with the high-pressure homogenization method is 250W; controlling the ultrasonic temperature below 20 ℃; the dialysis time is 2 h; homogenizing under high pressure at 150 MPa for 3 times, each for 1.5 min.
As a preferred scheme, the double-drug-loading carrier-free nano-particles can be added with a proper amount of freeze-drying protective agent for freeze-drying.
Further, the lyoprotectant can be one or more of lactose, sucrose, trehalose, glycine, mannitol, poloxamer and polyvinylpyrrolidone, and the amount of the lyoprotectant is 0.5% ~ 10% (w/v).
The invention also requires the application of the double-drug-loading carrier-free nano particle in treating tumor diseases.
Compared with the prior art, the invention has the characteristics and beneficial effects that:
1. the invention innovatively provides a double-drug-loading carrier-free nanoparticle integrating phototherapy and chemotherapy, which is assembled by only utilizing the interaction between two drug molecules, such as pi-pi conjugation, electrostatic attraction and hydrophobic interaction, so that the dependence on a carrier material is eliminated, the toxic and side effects caused by the introduction of a synthetic carrier are avoided, and the double-drug-loading carrier-free nanoparticle has good biocompatibility and biological safety;
2. the medicine is a typical medicine in phototherapy and chemotherapy, and the two medicines are innovatively combined by a nanotechnology, so that the bioavailability of the two hydrophobic medicines is greatly improved, the synergetic anti-tumor effect of the photo-chemotherapy is realized, and the medicine has potential clinical application value.
Drawings
Fig. 1 is a particle size diagram of a dual drug-loaded carrier-free nanoparticle prepared according to example 1 of the present invention and a particle size diagram of a nanoparticle added with a lyoprotectant for lyophilization and redissolution.
Fig. 2 is an observation result of a scanning electron microscope experiment of double drug-loaded carrier-free nanoparticles performed according to example 2.
Fig. 3 is a graph comparing the release curves of dual drug-loaded, carrier-free nanoparticles and single-drug injections according to example 3.
Fig. 4 shows the result of the cytotoxicity experiment of the double-drug-loaded carrier-free nanoparticles and the single-drug injection on 4T1 cells according to example 4.
Fig. 5 shows the results of the anti-tumor efficacy experiment of the dual drug-loaded carrier-free nanoparticles and the single injection solution on 4T1 tumor-bearing mice according to the present invention in example 5.
Fig. 6 is a graph showing the therapeutic results of the double drug-loaded carrier-free nanoparticles and single injection solution of the present invention on 4T1 cell tumor-bearing mice according to example 5.
Detailed Description
The invention is further described with reference to the following figures and detailed description. It should be noted that the following description is only for explaining the present invention, and does not limit the contents thereof.
Example 17-preparation of ethyl-10-hydroxycamptothecin (SN 38)/chlorin e6 (Ce 6) double drug loaded carrier-free nanoparticles (SN 38/Ce6 NPs): accurately weighing 7.6 mg of SN38 powder and 3.8 mg of Ce6, dissolving in 1 mL of DMF together, and performing ultrasonic dissolution to form an organic phase; dropwise adding the organic phase into 15 mL of deionized water under the ultrasonic condition of 250W, controlling the temperature to be below 20 ℃, continuously performing ultrasonic treatment for 10min, and dialyzing to remove an organic solvent (namely a mixed solution obtained by mixing the organic phase and the water phase); homogenizing under 150 MPa for 3 times (each for 1.5 min) to obtain SN38/Ce6 NPs. Dynamic Light Scattering (DLS) measures the particle size. Adding 1% (w/v%) of lactose as a freeze-drying protective agent into the SN38/Ce6NPs solution, and freeze-drying for 12 h to obtain SN38/Ce6NPs freeze-dried powder. The SN38/Ce6NPs freeze-dried powder is re-dissolved by water or 5 percent glucose. DLS measures the particle size.
The particle size of SN38/Ce6NPs prepared in example 1 is shown in figure 1, and as can be seen from figure 1, the average particle size of SN38/Ce6NPs is about 150 nm, and the particle size distribution is unimodal. After the freeze-drying protective agent lactose is added for freeze drying, water or 5% glucose (glucose solution with the mass fraction of 5%) is redissolved, and the grain diameter of the nanoparticle after redissolution has no obvious difference with that before freeze-drying. Indicating that the stability is good.
Example 2 dual drug loaded, unsupported nanoparticle morphology was observed using Scanning Electron Microscopy (SEM): after the SN38/Ce6NPs prepared in example 1 were freeze-dried without adding a freeze-drying protective agent, the freeze-dried powder was fixed on a copper column with a conductive adhesive, and gold-sprayed for 240 s under vacuum and 30 mA current. And (5) observing under a scanning electron microscope.
The SN38/Ce6NPs observed in example 2 are in a regular rod-shaped structure (figure 2), the lengths and the diameters of nanorods are relatively consistent, and the average length is 250 nm and the diameter is about 20 nm. Since DLS measurement assumes that the particles in solution are spherical, the measured particle size is the equivalent volume particle size. There is a slight deviation between SEM and DLS results.
Example 3 in vitro release profile of dual drug-loaded, carrier-free nanoparticles was determined by dynamic membrane dialysis: three batches of SN38/Ce6NPs are prepared according to example 1, SN38 injection and Ce6 injection (self-made by alkalized water) are used as controls, deionized water is diluted until the concentration of SN38 is 25 mug/mL and the concentration of Ce6 is 12.5 mug/mL, and an in-vitro drug release test is carried out. Precisely sucking 2 mL of the drug-containing solution, placing into a dialysis bag (MWCO = 8000-14000) soaked in distilled water, fastening the bag opening, adding 100mL of 0.01M PBS (containing 1% SDS), and releasing in a 37 ℃ water bath constant temperature oscillator (100 rpm). 5 mL were sampled at preset time points (0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 h) with addition of an isothermal, equal volume of release medium. And calculating the cumulative drug release percentage according to the measured drug concentration, and drawing a drug release curve.
The in vitro release results of SN38/Ce6NPs measured in example 3 are shown in FIG. 3, and it can be seen from FIG. 3 that for the homemade injection, the release behavior is complete passive diffusion through membrane, and the release is complete within 6 h. On the contrary, SN38/Ce6NPs can be slowly released for about 24 h, and SN38 and Ce6 in the nanoparticles are in a sequential release state, namely, Ce6 is released quickly, SN38 is released slowly, and 24 h reaches an equilibrium state. The slow release of nanoparticles is presumed to help prolong the circulation time of nanoparticles in blood based on the correlation between in vitro release and in vivo bioavailability.
Example 4 determination of the in vitro cytotoxic effect of double drug-loaded carrier-free nanoparticles by MTT method: SN38/Ce6NPs were prepared as in example 1. Culturing 4T1 cells to logarithmic phase, inoculating to 96-well plate at 10000/well, 37 deg.C, 5% CO2Culturing for 12 h. SN38/Ce6NPs and the self-made injection are diluted by a culture medium without fetal bovine serum, and then added into a 96-well plate, each concentration is 6 duplicate wells, and a blank culture medium is used as a control. Wherein, the laser irradiation with 660 nm wavelength is 5 min (laser power is 5 mW) 4 h after the administration of the Ce6 injection group and the SN38/Ce6NPs group. The 96-well plate is put into an incubator to be continuously cultured for 48 hours, and then the supernatant is discarded. Adding 20 mu L of 5 mg/mL MTT solution into each hole, and continuously incubating for 4 h in the incubator. After upper layer liquid of each well is carefully discarded, 150 muL of DMSO is added into each well, and the culture plate is placed on a microplate oscillator to oscillate for 10min, so that crystals are dissolved. The OD value at 490 nm was measured on a microplate reader, and the cell viability was calculated according to the following formula (%) = (drug group OD mean value/blank group OD mean value) × 100%. The drug concentration is used as the abscissa and the survival rate is used as the ordinate to draw a dose-effect curve.
The in vitro cytotoxicity results of SN38/Ce6NPs measured in example 4 are shown in FIG. 4, and it can be seen from FIG. 4 that the inhibition effects of SN38 injection, Ce6 injection and SN38/Ce6NPs on 4T1 cells are concentration-dependent, and compared with the injection, the killing effect of SN38/Ce6NPs on cells is obviously improved. This is probably because, SN38 in SN38/Ce6NPs is almost all lactone-type, and the proportion of lactone-type in SN38 injection is small, and only lactone-type drugs can exert antitumor activity; meanwhile, SN38/Ce6NPs realize the synergistic effect of SN38 and Ce6 chemotherapy-phototherapy, and the cytotoxic effect on 4T1 cells is obviously improved compared with that of a single medicament.
Examples5, determining the in-vivo drug effect of the double-drug-loading carrier-free nanoparticle by using a 4T1 tumor-bearing mouse model: SN38/Ce6NPs were prepared as in example 1. 4T1 tumor-bearing mice(s) (subcutaneously inoculated) were randomly divided into 5 groups of 6 mice each, and administered by tail vein injection every 2 days for 10 days in addition to normal diet. Blank group was given 0.2 mL of physiological saline; the injection group is used for tail vein injection of SN38/Ce6NPs and self-made injection according to the dosage of SN386 mg/kg and Ce 63 mg/kg. Wherein, the laser with the wavelength of 660 nm irradiates the tumor part for 10min (the laser power is 5 mW) 4 h after the Ce6 injection group and the SN38/Ce6NPs group are administrated. Mice were observed daily for loss of hair color, activity, sleep and death. And tumor size was measured with a vernier caliper and tumor volume was calculated according to the following formula: tumor volume V (mm)3）= 0.5×L×W2. Wherein, L and W are the length and width of the tumor, respectively. After the experiment was completed, the mice were sacrificed and tumors of each group were taken out and photographed.
The in vivo efficacy of SN38/Ce6NPs measured in example 5 is shown in FIG. 5 and FIG. 6, and as can be seen from the time-dependent change curve of tumor volume of each group of mice in FIG. 5, the tumor volume of mice in saline group increases to 1700mm at the end of the experiment3And the tumor volume of mice in each experimental group is obviously reduced compared with that of the normal saline group: the tumor volume of the mice of the SN38 injection group and the Ce6 injection group is increased to 1100 mm3Left and right; the tumor volume of mice in the group without adding laser SN38/Ce6NPs is increased to 450mm3Left and right; the tumor volume of mice in the group of SN38/Ce6NPs is increased to 350 mm by adding laser3Left and right. Compared with the injection, the SN38/Ce6NPs group tumor inhibition effect is obviously improved (P)<0.05), which is also probably because the higher proportion of lactone type SN38 in SN38/Ce6NPs plays stronger anti-tumor activity, and the synergistic effect of the photo-chemotherapy double drugs in the nanoparticles obviously improves the tumor inhibition effect. The tumor inhibition effect of the group without adding laser SN38/Ce6NPs is also obvious, which is probably because the nanoparticles with the particle size of about 150 nm can be passively targeted to the tumor part through the EPR effect and are taken up by tumor cells with higher cell uptake efficiency, thereby playing a better role in inhibiting the growth of the tumor. The tumor picture results of the mice in each group in FIG. 6 are substantially consistent with the results of the tumor volume change curves with timeAs can be seen from the pictures, the tumors of the mice in the SN38/Ce6NPs group become smaller obviously, and the effect of the nanoparticle group irradiated by laser is more obvious.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (9)
1. The double-drug-loading carrier-free nanoparticle is characterized by being formed by assembling drug intermolecular interaction between phototherapeutic drugs and chemotherapeutic drugs, wherein the interaction between the drug molecules comprises pi-pi conjugation, electrostatic attraction and hydrophobic interaction.
2. The dual drug-loaded, carrier-free nanoparticle of claim 1, wherein the phototherapeutic agent is a drug with photosensitizer property, the drug with photosensitizer property is chlorin e6, the chemotherapeutic agent is a camptothecin drug, and the camptothecin drug is camptothecin, 10-hydroxycamptothecin or 7-ethyl-10-hydroxycamptothecin.
3. The dual drug-loaded carrier-free nanoparticle according to claim 2, wherein the molecular molar ratio of the photosensitizer property drug to the camptothecin class drug is 1:1, 1:2, 1:3 or 1: 4.
4. The double-drug-loaded carrier-free nanoparticle according to claim 1, wherein the particle size of the double-drug-loaded carrier-free nanoparticle is 170 nm, the double-drug-loaded carrier-free nanoparticle is lyophilized by adding a proper amount of a lyoprotectant, the lyoprotectant is one or more of lactose, sucrose, trehalose, glycine, mannitol, poloxamer and polyvinylpyrrolidone, and the amount of the lyoprotectant is 0.5% ~ 10% w/v.
5. A preparation method of double-drug-loading carrier-free nanoparticles, which is based on the double-drug-loading carrier-free nanoparticles of any one of claims 1 to 4, and is characterized in that the double-drug-loading carrier-free nanoparticles are prepared by combining an anti-solvent precipitation method with a high-pressure homogenization method or combining an alkali-soluble acid precipitation method with a high-pressure homogenization method.
6. The preparation method according to claim 5, wherein the anti-solvent precipitation method is combined with a high-pressure homogenization method, and specifically comprises the following steps:
(1) co-dissolving a certain amount of phototherapeutic drug and chemotherapeutic drug in an organic solvent to form an organic phase;
(2) the organic phase is added into the water phase drop by drop under the ultrasonic condition;
(3) continuing to perform ultrasonic treatment for 10min at low temperature;
(4) dialyzing to remove the aqueous phase and the organic phase;
(5) homogenizing for several times under high pressure to obtain double-drug-loading carrier-free nanoparticles.
7. The preparation method according to claim 5, wherein the alkali-dissolution acid-precipitation method is combined with a high-pressure homogenization method, and the method comprises the following specific steps:
(1) dissolving a certain amount of phototherapeutic drugs and chemotherapeutic drugs in 10 mL of 0.2mol/L NaOH solution to obtain a mixed solution;
(2) dropwise adding 0.2mol/L HCl solution into the mixed solution in the step (1) under ultrasonic conditions to reach the pH = 5;
(3) continuing to perform ultrasonic treatment for 10min at low temperature;
(4) centrifuging at 10000 rpm for 10min, discarding supernatant, and resuspending the precipitate in deionized water;
(5) homogenizing for several times under high pressure to obtain double-drug-loading carrier-free nanoparticles.
8. The preparation method according to claim 6, wherein in the step (1), the organic phase is one or two of DMF or DMSO, the aqueous phase is deionized water or a glucose solution with a glucose mass fraction of 5%, and the volume ratio of the organic phase to the aqueous phase is 1:10 ~ 1: 15.
9. The preparation method according to claim 6 or 7, wherein ultrasonic power in the combination of the anti-solvent precipitation method and the high-pressure homogenization method or the combination of the alkali-dissolution acid precipitation method and the high-pressure homogenization method is 250W; the ultrasonic temperature is controlled below 20 ℃; the dialysis time is 2 h; homogenizing under 150 MPa for 3 times, each for 1.5 min.
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| CN111558043A (en) * | 2020-06-03 | 2020-08-21 | 哈尔滨工业大学 | Preparation method of terpene micromolecule assembled redox response photosensitive drug |
| CN111632144A (en) * | 2020-07-06 | 2020-09-08 | 聊城大学 | High-drug-loading-rate photo-chemotherapy bifunctional nanoparticles and preparation method thereof |
| CN113350503A (en) * | 2021-05-20 | 2021-09-07 | 沈阳药科大学 | Carrier-free hybrid nano assembly and preparation method and application thereof |
| CN113384698A (en) * | 2021-06-09 | 2021-09-14 | 福州大学 | Self-assembled nano-medicament for synergetic chemotherapy/acousto-photodynamic therapy and application thereof |
| CN116327706A (en) * | 2023-02-01 | 2023-06-27 | 中山大学附属第八医院（深圳福田） | Nanometer drug delivery system and preparation method and application thereof |
| CN117122681A (en) * | 2023-08-08 | 2023-11-28 | 广东药科大学 | Carrier-free self-assembled drug nano particle and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104274413A (en) * | 2014-07-25 | 2015-01-14 | 中国医学科学院药用植物研究所 | Nanoparticles of camptothecin drugs and preparation method of nanoparticles |
| CN108159422A (en) * | 2016-12-07 | 2018-06-15 | 江西科维协同创新药物有限公司 | A kind of preparation method of self assembly drug-loading system and its compound formulation |
-
2019
- 2019-08-20 CN CN201910769983.8A patent/CN110665003A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104274413A (en) * | 2014-07-25 | 2015-01-14 | 中国医学科学院药用植物研究所 | Nanoparticles of camptothecin drugs and preparation method of nanoparticles |
| CN108159422A (en) * | 2016-12-07 | 2018-06-15 | 江西科维协同创新药物有限公司 | A kind of preparation method of self assembly drug-loading system and its compound formulation |
Non-Patent Citations (2)
| Title |
|---|
| YAN WEN等: ""Carrier-free, self-assembled pure drug nanorods composed of 10-hydroxycamptothecin and chlorin e6 for combinatorial chemophotodynamic antitumor therapy in vivo"", 《NANOSCALE》 * |
| 杨晓凤等: ""羟基喜树碱纳米晶处方工艺筛选及工业化初探"", 《医药导报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111558043A (en) * | 2020-06-03 | 2020-08-21 | 哈尔滨工业大学 | Preparation method of terpene micromolecule assembled redox response photosensitive drug |
| CN111632144A (en) * | 2020-07-06 | 2020-09-08 | 聊城大学 | High-drug-loading-rate photo-chemotherapy bifunctional nanoparticles and preparation method thereof |
| CN111632144B (en) * | 2020-07-06 | 2022-07-29 | 聊城大学 | High-drug-loading-rate photo-chemotherapy bifunctional nanoparticles and preparation method thereof |
| CN113350503A (en) * | 2021-05-20 | 2021-09-07 | 沈阳药科大学 | Carrier-free hybrid nano assembly and preparation method and application thereof |
| CN113350503B (en) * | 2021-05-20 | 2022-12-09 | 沈阳药科大学 | Carrier-free hybrid nano assembly and preparation method and application thereof |
| CN113384698A (en) * | 2021-06-09 | 2021-09-14 | 福州大学 | Self-assembled nano-medicament for synergetic chemotherapy/acousto-photodynamic therapy and application thereof |
| CN116327706A (en) * | 2023-02-01 | 2023-06-27 | 中山大学附属第八医院（深圳福田） | Nanometer drug delivery system and preparation method and application thereof |
| CN117122681A (en) * | 2023-08-08 | 2023-11-28 | 广东药科大学 | Carrier-free self-assembled drug nano particle and preparation method and application thereof |
| CN117122681B (en) * | 2023-08-08 | 2024-03-15 | 广东药科大学 | Carrier-free self-assembled drug nano particle and preparation method and application thereof |
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