CN111388671A - Nano-drug carrier, drug-carrying system containing nano-drug carrier and preparation method of drug-carrying system - Google Patents
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Abstract
The invention discloses a nano drug-loading system with high drug-loading capacity, high photothermal conversion rate and multi-factor response drug release function, which adopts synthesized nano hollow CuS (copper sulfide), obtains CuS-PEG (copper sulfide-polyethylene glycol) through the combined action load of metal sulfide and amino, loads anticancer drug DOX (adriamycin) through electrostatic adsorption and a CuS hollow structure, and obtains a CuS-PEG-DOX @ HA type nano drug carrier through the combined load of chemical bonds and HA.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a nano-medicine carrier with high medicine-loading capacity, high photothermal conversion rate and multi-factor response medicine release function, a medicine-loading system containing the nano-medicine carrier and a preparation method of the medicine-loading system.
Background
Malignant tumors caused by disordered cell division have seriously threatened human health. And these cells grow unlimitedly beyond the cell boundaries and have the ability to invade adjacent tissues and spread to other organs. Although the technology for diagnosing and treating cancer is rapidly developed, many types of cancer cannot be cured at present. Chemotherapy is currently one of the most important therapeutic approaches for the treatment of tumors. However, due to the serious toxic and side effects of chemotherapy drugs, the design of drug carriers for chemotherapy drugs is a popular research direction in pharmacy in recent years. These drug carriers achieve drug delivery through specific design and are expected to deliver drugs to specific target sites to achieve therapeutic effects while minimizing toxic side effects to normal cells or tissues with the greatest possible potential.
The nano-drug carrier is a novel carrier with the particle size of 10-1000 nm. The particle size of the nano-drug carrier is far smaller than that of a capillary vessel passage, and the nano-drug carrier has the advantages of reducing the toxic and side effects of drugs, improving the stability of the drugs, slowly releasing and controlling the drugs, realizing the targeted release of the drugs and the like, and has extremely wide application in the field of medicine.
The first staged and breakthrough goal of cancer therapy is to selectively kill tumor cells while avoiding damage to surrounding healthy tissue. Recently, significant attention has been paid to the development of photothermal therapy. Because the light can be easily focused and adjusted to achieve local treatment, damage to healthy tissue is also relatively small. Furthermore, in the course of using light-induced treatment modalities, the dosing regimen (e.g., treatment area, time, and dose) can be adjusted as needed according to the patient's physiological response and clinical needs, and thus photothermal treatment modalities have been identified as non-invasive, direct, and accurate cancer treatment approaches.
Photothermal therapy utilizes a photothermal agent (also referred to as a light absorber) to convert the energy of near infrared light into heat, killing cancer cells at a specific target site, and develops rapidly due to the good biosafety of the photothermal therapy mode and the only slight attenuation of near infrared excitation light in tissues. In addition, photothermal therapy overcomes the disadvantages of conventional hyperthermia (external hyperthermia) that is not selective and causes severe side effects. The maximum temperature gradient is produced at the body surface due to the conventional high temperature and most of the energy is dissipated in the external radiation path located in healthy tissue. In contrast, photothermal therapy has the ability to concentrate the energy of the near infrared light on the tumor, causing local thermal damage, reversing the direction of heat loss (inside-out hyperthermia) and minimizing side effects on healthy tissue. Therefore, in order to overcome the inhomogeneous distribution of heat, increase the efficiency of photothermal conversion and correspondingly increase the thermal death of subcutaneous tumors, a number of photothermal agents have been extensively explored, especially nanomaterials with strong absorption in the NIR region (700-1000 nm).
In recent years, a plurality of drug carrier materials with photo-thermal conversion performance are developed successively, and the nano materials can effectively absorb light irradiated from the outside and convert the light into heat, thereby effectively combining photo-thermal therapy and chemical drug therapy. Meanwhile, in the design and preparation of the drug carrier material, various hidden dangers exist, and the future development and application of the drug carrier material can be seriously influenced. These concerns are as follows:
on one hand, some drug carrier materials with photothermal conversion performance have poor biocompatibility, may cause adverse reactions in vivo, and may cause serious damage to normal cells and tissues of an organism during use. Meanwhile, some materials have the problems of poor photo-thermal stability, low photo-thermal conversion efficiency and the like, and the purpose of effective and continuous photo-thermal treatment cannot be achieved.
On the other hand, some drug carrier materials have insufficient drug loading capacity and low drug loading capacity, which can seriously affect the chemotherapy effect, and the dosage needs to be increased when in use, thereby greatly affecting the biological safety of the chemotherapy. Moreover, some drug carrier materials do not have the specificity to cancer tissues, and the enrichment of the materials at the tumor tissues is difficult to realize, so that the killing effect of the drug carrier materials on the tumor tissues is greatly reduced, chemotherapeutic drugs can be diffused to various normal physiological tissues through normal physiological circulation, the same toxic effect is generated on normal cells and tissues, the larger chemotherapeutic side effect is generated on organisms, and the biological safety of the organisms is seriously influenced. Meanwhile, in some drug carrier materials, the release capacity of the loaded chemotherapeutic drug is difficult to regulate and control through in vivo and in vitro environments, uncontrollable drug release is easy to occur at non-tumor tissues in organisms, and great toxic and side effects are generated and the effect of chemical drug treatment is influenced.
The copper sulfide nanoparticles, which are often used as catalysts, can generate surface plasmon resonance under the irradiation of near infrared light to realize efficient photothermal conversion, so that the copper sulfide nanoparticles are widely applied to photothermal treatment of tumors.
Disclosure of Invention
The invention aims to provide a nano-drug carrier with high drug loading capacity, high photothermal conversion rate and multi-factor response drug release function, a drug loading system containing the nano-drug carrier and a preparation method of the drug loading system.
In order to achieve the above object, the technical means adopted by the present invention are as follows:
the nano-drug carrier is characterized in that the nano-drug carrier is a composite carrier material formed by self-assembling polyethylene glycol modified nano-hollow copper sulfide and hyaluronic acid.
As a preferable mode, the polyethylene glycol modified nano hollow copper sulfide is obtained by loading nano hollow copper sulfide and folic acid polyethylene glycol amino through the combination action of metal sulfide and amino.
As a preferred mode, the preparation process of the polyethylene glycol modified nano hollow copper sulfide is as follows: dissolving the folic acid polyethylene glycol amino in deionized water, adding a CuS material, and carrying out ultrasonic treatment, centrifugation, precipitation, cleaning and drying.
As a preferable mode, the preparation process of the nano hollow copper sulfide is as follows: adding CuCl2Dissolving in deionized water, adding PVP, adjusting the pH of the solution to 9.0 by using NaOH, and adding hydrazine hydrate under the condition of continuous stirring; mixing Na2And S is dissolved in deionized water, the prepared solution is slowly added, and after uniform mixing, the temperature is raised for reaction, cooling, centrifugal precipitation, cleaning and drying are carried out.
A drug-carrying system comprises the nano-drug carrier and a drug loaded by the nano-drug carrier.
In a preferable mode, the drug loaded on the nano drug carrier is adriamycin.
The method for preparing the medicine carrying system is characterized by comprising the following steps:
s1: dissolving the medicine in a buffer solution, mixing with CuS-PEG, centrifuging, precipitating, cleaning, drying, and loading the medicine on polyethylene glycol modified nano hollow copper sulfide;
s2: dissolving EDC and NHS in deionized water, adding hyaluronic acid, mixing, adding polyethylene glycol-modified nano hollow copper sulfide loaded with medicine, mixing, centrifuging, precipitating, cleaning and drying.
In the invention, nano-scale hollow CuS (copper sulfide) is synthesized, the material HAs higher photo-thermal conversion capability and good photo-thermal stability, CuS-PEG (copper sulfide-polyethylene glycol) is obtained by loading through the combined action of metal sulfide and amino, the biocompatibility of the material is improved, the material HAs extremely high drug loading capability through electrostatic adsorption and loading of anticancer drug DOX (adriamycin) on the CuS hollow structure, the CuS hollow structure is combined with loaded HA through chemical bonds to obtain a CuS-PEG-DOX @ HA type nano drug carrier,
compared with the prior art, the invention has the beneficial effects that: the obtained CuS-PEG-DOX @ HA type nano-drug carrier HAs specific targeting property on cancer cells. Meanwhile, the drug release capacity of the nano drug carrier has high controllability, can be simultaneously influenced by the pH environment in an organism, the hyaluronidase environment and the near infrared light irradiation environment outside the organism, and has high specificity and biological safety.
Drawings
FIG. 1 is a representation of the morphology and structure of nano-scale hollow CuS prepared by a liquid phase method, wherein (a) is a SEM representation; (b) TEM representation; (c) XRD characterization; (d) HRTEM characterization.
FIG. 2 shows the particle size distribution of CuS and CuS-PEG in deionized water.
FIG. 3 is a suspension property test chart of CuS and CuS-PEG.
FIG. 4 shows the loading of CuS and CuS-PEG.
FIG. 5 is a representation of a CuS-PEG-DOX @ HA material, wherein (a) a TEM image of the CuS-PEG-DOX @ HA material; (b) FTIR plots of different materials.
Fig. 6 is a photo-thermal characterization of the material.
Detailed Description
The present invention is aimed at overcoming the defects of the prior art and providing a nano-drug carrier material with high drug loading capacity, high photothermal conversion rate and multi-factor response drug release function, and the present invention is further described in detail with reference to the following examples.
Examples
The drug-carrying system carrying adriamycin (DOX) is prepared by the following steps:
(1) 85.24mg of CuCl2Dissolving in 250m L deionized water, adding 240mg PVP, adjusting the pH of the solution to 9.0 by NaOH, adding 64 mu L hydrazine hydrate under continuous stirring, and stirring at normal temperature for 5 min.
(2) 120 mg of Na2And dissolving S in 250m L deionized water, slowly adding the solution prepared in the previous step, uniformly mixing, heating to 75 ℃, reacting for 2 hours at 75 ℃, and naturally cooling to room temperature.
(3) And centrifuging the reacted solution at a centrifugal speed of 8000r/min, washing the separated precipitate for 3 times by using deionized water, putting the precipitate into an oven, and drying the precipitate for 4 hours at the temperature of 60 ℃ to obtain the CuS material.
The nanoscale hollow CuS is prepared by a liquid phase method, and the appearance and the structural representation of the nanoscale hollow CuS are shown in figure 1. The characteristics of SEM and TEM show that the prepared material is hollow spherical and has the diameter of about 200nm, and the characteristics of XRD and HRTEM show that the prepared material is a CuS material and has no obvious impurities.
(4) 20mg of Folate-PEG-NH was taken2Dissolving (folic acid polyethylene glycol amino) in 40m L deionized water, adding 40mg CuS material, performing ultrasonic treatment for 4h, centrifuging the solution after reaction at the centrifugal speed of 8000r/min, washing the separated precipitate with deionized water for 3 times, putting the precipitate into an oven, and drying at 60 ℃ for 4h to obtain the CuS-PEG material.
By reaction of CuS with-NH2Binding of (b) A salt-PEG-NH2Loaded on CuS, the performance characteristics of the material are shown in figure 2. The suspension property representation of the CuS and CuS-PEG materials in deionized water is obtained by carrying out suspension property test on the CuS and CuS-PEG materials. The test results are shown in fig. 3: after the PEG is loaded, the suspension property of the material is obviously improved, and the material is not easy to aggregate and precipitate in deionized water, which proves that the Folate-PEG-NH2Successfully combines with the CuS material to successfully synthesize the CuS-PEG material.
(5) And (2) dissolving 20mg of DOX in 80m L PBS solution, adding 40mg of CuS-PEG, stirring overnight at room temperature in a dark place, centrifuging the solution at a centrifugal speed of 8000r/min, washing the separated precipitate for 3 times by using deionized water, putting the washed precipitate into an oven, and drying for 4 hours at the temperature of 60 ℃ to obtain the CuS-PEG-DOX material.
DOX can be loaded on the prepared drug carrier material by utilizing the hollow structure of CuS, and the drug loading rate is shown in figure 4. By comparing the drug loading of the two materials CuS and CuS-PEG, it can be seen that: CuS and CuS-PEG both have extremely high drug loading capacity, and no obvious influence is caused on the drug loading capacity after the PEG is loaded, because the DOX can be effectively loaded on the material due to the unique hollow structure of the material, the material is proved to have extremely high drug loading capacity.
(6) 30mg of EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) and 45mg of NHS (N-hydroxysuccinimide) are dissolved in 40m L deionized water, 20mg of HA (hyaluronic acid) is added, after stirring for 1h at room temperature, 30mg of CuS-PEG-DOX is added, after stirring for 2h at room temperature, the solution is centrifuged at the centrifugal speed of 8000r/min, and the separated precipitate is washed for 3 times by deionized water, then is placed in an oven and is dried for 4h at the temperature of 60 ℃, thus obtaining the CuS-PEG-DOX @ HA material.
HA can be loaded on the surface of the material by utilizing the reaction of carboxyl on the Folate-PEG-NH2 and amino on the HA, so that the CuS-PEG-DOX @ HA type material is synthesized, and the cancer cell targeting property and the drug release controllability of the material are improved. The CuS-PEG-DOX @ HA material is characterized as shown in FIG. 5. As can be seen from TEM picture, after HA is loaded, a layer of membranous substance appears on the CuS surface, and as can be seen from FTIR picture, the CuS-PEG-HA group is 3413 nm-1Has strong absorption peak due to the absorption peak of stretching vibration of OH and NH of hyaluronic acid, 1644 nm-1The strong absorption peak is due to the absorption of O = C-N functional group of hyaluronic acid, 1051nm-1The photothermal property of the CuS-PEG-DOX @ HA type material is characterized, and the result is shown in FIG. 6, FIG. 6 (a) is a temperature rise curve of samples with different CuS concentrations under the action of 1.8W near infrared light at 808 nm, wherein the higher the concentration is, the more obvious the temperature rise effect is, because the number of atoms vibrated by the excitation of the near infrared light is increased, and the temperature is increased more rapidly, in the a diagram, the temperature rise of an experimental group with the lowest concentration is more than 10 ℃, the normal body temperature of a human body is about 37 ℃, the 10 ℃ is increased to 47 ℃, and 42 ℃ is the highest tolerance temperature of cancer cells, and normal cells can tolerate 1h at 48 ℃, so that the visible light carrier with the CuS as the substrate can meet the photothermal property requirement of medical phototherapy, FIG. 6 (b) is the photothermal property requirement that the CuS solution with 1mg/m L is under the irradiation of different wattages at 808 nm, the curve of the CuS solution is shown, the higher the watt number is, the modification effect is, the more obvious the photothermal property is, and the more obvious the temperature rise effect is, and the photothermal stability of the prepared drug carrier is shown by the photothermal property of the temperature rise curve of the nano-PEG-temperature, and the photothermal property of the photothermal material is obviously improvedThe hollow spherical material has higher photo-thermal conversion rate and photo-thermal stability, and is an ideal photo-thermal agent required in photo-thermal therapy. Meanwhile, the material has extremely high drug-loading rate, can achieve higher drug concentration through smaller drug dosage, and is an excellent carrier for the chemical drug therapy. The material combines photothermal therapy and chemical drug therapy, and has extremely wide application prospect;
the invention is well implemented in accordance with the above-described embodiments. It should be noted that, based on the above structural design, in order to solve the same technical problems, even if some insubstantial modifications or colorings are made on the present invention, the adopted technical solution is still the same as the present invention, and therefore, the technical solution should be within the protection scope of the present invention.
Claims (7)
1. The nano-drug carrier is characterized in that the nano-drug carrier is a composite carrier material formed by self-assembling polyethylene glycol modified nano-hollow copper sulfide and hyaluronic acid.
2. The nano-drug carrier according to claim 1, wherein the polyethylene glycol modified nano-hollow copper sulfide is obtained by loading nano-hollow copper sulfide and folic acid polyethylene glycol amino through the combination of metal sulfide and amino.
3. The nano-drug carrier according to claim 3, wherein the polyethylene glycol modified nano-hollow copper sulfide is prepared by the following steps: dissolving the folic acid polyethylene glycol amino in deionized water, adding a CuS material, and carrying out ultrasonic treatment, centrifugation, precipitation, cleaning and drying.
4. The nano-drug carrier according to claim 3, wherein the nano-hollow copper sulfide is prepared as follows: adding CuCl2Dissolving in deionized water, adding PVP, adjusting the pH of the solution to 9.0 by using NaOH, and adding hydrazine hydrate under the condition of continuous stirring; mixing Na2S is dissolved in deionized water and slowly added to the solutionAnd uniformly mixing the solution, heating for reaction, cooling, centrifugally precipitating, cleaning and drying.
5. A drug carrier system, which is characterized in that the drug carrier system comprises the nano drug carrier of any one of claims 1 to 4 and a drug loaded on the nano drug carrier.
6. The drug delivery system of claim 5, wherein the drug carried by the nano-drug carrier is doxorubicin.
7. A method for preparing the drug delivery system of claims 5-6, comprising the steps of:
s1: dissolving the medicine in a buffer solution, mixing with CuS-PEG, centrifuging, precipitating, cleaning, drying, and loading the medicine on polyethylene glycol modified nano hollow copper sulfide;
s2: dissolving EDC and NHS in deionized water, adding hyaluronic acid, mixing, adding polyethylene glycol-modified nano hollow copper sulfide loaded with medicine, mixing, centrifuging, precipitating, cleaning and drying.
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| CN113082213A (en) * | 2021-03-10 | 2021-07-09 | 武汉科技大学 | Hollow copper sulfide nano-drug carrier, preparation method and application |
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