CN112999169B - Regorafenib-coated nano porous hydroxyapatite sustained-release microsphere as well as preparation method and application thereof - Google Patents

Regorafenib-coated nano porous hydroxyapatite sustained-release microsphere as well as preparation method and application thereof Download PDF

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CN112999169B
CN112999169B CN202110231978.9A CN202110231978A CN112999169B CN 112999169 B CN112999169 B CN 112999169B CN 202110231978 A CN202110231978 A CN 202110231978A CN 112999169 B CN112999169 B CN 112999169B
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于凤波
孟焕
高俊霞
王强
崔燎
黄春明
张治平
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Abstract

The invention relates to the field of drug sustained release preparations, in particular to a regorafenib coated nano porous hydroxyapatite sustained release microsphere and a preparation method and application thereof. The slow release microspheres comprise 5-10 parts of regorafenib and 10-20 parts of nano porous hydroxyapatite; the nano-porous hydroxyapatite is modified by polyglutamic acid, polyaspartic acid or polylysine. According to the invention, through carrying out polyamino acid modification on the nano-porous hydroxyapatite, the release rate of the active ingredient regorafenib can be effectively controlled, the drug release rate and the treatment effect of the drug are improved, the side effect of the drug on a patient can be remarkably reduced, and the nano-porous hydroxyapatite carrier has a wide application prospect in the field of drug sustained release carriers.

Description

Regorafenib-coated nano porous hydroxyapatite sustained-release microsphere as well as preparation method and application thereof
Technical Field
The invention relates to the field of drug sustained release preparations, in particular to a regorafenib coated nano porous hydroxyapatite sustained release microsphere and a preparation method and application thereof.
Background
Cancer is one of the major factors threatening human life in modern society. The world cancer report issued by the world health organization international research center for cancer (IARC)2014 predicts that global cancer cases will exhibit a rapidly growing situation, increasing year by year from 1400 to 2012 to 1900 to 2025 to 2400 by year 2035. Under such severe conditions, it is very important to find an effective treatment for cancer diseases. In the field of clinical medicine, although the traditional anticancer drug injection therapy is convenient and quick and has a certain treatment effect, the injury can be caused due to overlarge local drug concentration in a body, the drug molecule metabolism of the human body is quick, the effective drug concentration is short in the retention time in the body, the pain of diseases can be temporarily relieved, the long-term repeated administration of the drug to a patient is needed, the accumulated residual quantity of the drug molecules in the human body is continuously increased, and certain toxic and side effects are caused to the patient. In order to control the release of drugs, research on the sustained release performance of drugs carried by drug carriers has received much attention from researchers at home and abroad. In the process of constructing a Drug release System (Drug release System), the Drug carrier is used as an important component of the Drug release System, and physicochemical factors such as morphology particle size, chemical components, pore structure and the like can influence the Drug loading performance of the carrier and the Drug release performance of the Drug release System, so that ideal release effect and treatment effect are achieved. Therefore, continuous research and innovation on the preparation method of the drug carrier material become new research hotspots in recent years.
Hydroxyapatite (HAP) is a major inorganic component of human bones and teeth, and is widely used in the research of drug sustained-release carriers due to its excellent biocompatibility and biosafety. Studies have shown that HAP has good biocompatibility and little toxicity to normal cells. Meanwhile, HAP has good biodegradability, and the degradation period of the HAP is closely related to the physical and chemical properties of the material. The porous hollow HAP has a hollow structure and a porous structure, has the advantages of large specific surface area, good biocompatibility and the like, and has the advantages of high drug loading rate, high entrapment rate, small side effect and the like when being used as a drug slow release carrier, so that the drug slow release system has the advantages of good drug loading performance, good drug slow release performance and the like, and can safely, slowly and efficiently control the drug to be released as required. Regorafenib is a novel anticancer drug, the chemical structure of which is fat-soluble macromolecule, the solubility of which in water is not high, and the regorafenib is not beneficial to the loading of hydroxyapatite. The regorafenib-coated nano porous hydroxyapatite sustained release microspheres with high drug loading and long drug release time are prepared by modifying hydroxyapatite.
Disclosure of Invention
The invention provides a regorafenib-coated nano porous hydroxyapatite sustained release microsphere and a preparation method and application thereof. Through carrying out polyamino acid modification on the nano porous hydroxyapatite, the release rate of the active ingredient regorafenib can be effectively controlled, the drug release rate and the treatment effect of the drug are improved, the side effect of the drug on patients can be remarkably reduced, and the nano porous hydroxyapatite sustained-release carrier has wide application prospect in the field of drug sustained-release carriers.
In order to achieve the purpose, the invention adopts the technical scheme that:
a regorafenib-coated nano porous hydroxyapatite sustained release microsphere comprises the following components in parts by weight: 5-10 parts of regorafenib and 10-20 parts of nano porous hydroxyapatite; the nano-porous hydroxyapatite is modified by polyamino acid.
Preferably, the polyamino acid is polyglutamic acid, polyaspartic acid or polylysine.
Further, the invention also provides a preparation method of the regorafenib coated nano porous hydroxyapatite sustained release microspheres, which comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving surfactant in 0.5% toluene, mixing 70ml toluene and surfactant solution in 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 10-12 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2-3 hr to obtain product, separating, filtering, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and oven drying at 80 deg.C for 8-12 hr to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of solution is dripped into the suspension at the speed of 2 ml/min; adjusting the pH value of the solution by using 20 percent NaOH solution in percentage by mass under the conditions of normal pressure and constant temperature water bath at 60 DEG C10-12, separating a product after reacting for a certain time, washing with distilled water until the solution is neutral, washing with absolute ethyl alcohol for 3 times, and drying in an oven at 80 ℃ for 8-12 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: weighing 10.0g of amino acid monomer, placing the amino acid monomer in a 250ml round bottom flask, adding 1.0ml of 85% phosphoric acid solution and 2.0ml of distilled water, fully and uniformly mixing, gradually heating the mixture to 220 ℃ under the condition of oil bath, simultaneously vacuumizing, reacting for 1-2h, cooling to 80 ℃ of oil bath temperature after the reaction is finished, and adding 30ml of 80% ethanol water solution to completely dissolve the product; and slowly dripping 10ml of the dissolved solution into 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature under the condition of stirring, filtering and drying the resultant to obtain the slow-release microspheres.
Preferably, the surfactant in the step (1) is tween-80 or lecithin.
Preferably, the oven drying time in the steps (1) and (2) is 10 h.
Preferably, the oil bath heating temperature in the step (3) is 200 ℃.
Furthermore, the invention also provides application of the regorafenib-coated nano-porous hydroxyapatite sustained release microspheres in preparation of a cancer treatment drug.
Preferably, the cancer is colorectal cancer or liver cancer.
Regorafenib is an oral multi-kinase inhibitor developed by Bayer pharmaceutical companies, the structure of the drug is similar to that of sorafenib, and the difference is that only one fluorine atom is added on the central benzene ring of regorafenib, so that the two drugs are similar and have different biochemical characteristics. In vitro biochemical analysis shows that compared with sorafenib, regorafenib has stronger inhibition effect on VEGFR-2, PDGFR-beta, FGFR-1 and c-Kit. Meanwhile, regorafenib can also inhibit Tie-2 and has a wider anti-angiogenesis effect. Both regorafenib and sorafenib target c-RAF, wild-type BRAF and BRAFV600E in inhibiting tumor cell proliferation and interfere with the MAP kinase intracellular signaling pathway that is abnormally activated at tumorigenesis. Among them, inhibition of MAP kinase p38 is a unique feature of regorafenib. Therefore, compared with sorafenib, the regorafenib-targeted kinase has a wider range and a stronger pharmacological effect, and is a promising antitumor drug. The chemical structure of regorafenib is as follows:
Figure BDA0002958813900000031
compared with the prior art, the invention has the beneficial effects that:
(1) the invention uses nano-porous hydroxyapatite as a drug carrier to carry out load combination with a clinical antitumor drug regorafenib to obtain a drug carrier system. The nanotechnology is helpful for improving the solubility of the insoluble drug regorafenib and improving the utilization rate of the drug. The nano-drug delivery system can improve the membrane penetration capacity and stability of the drug and is also beneficial to improving the concentration of the drug at a lesion site. The hydroxyapatite is a complex process with physical adsorption and chemical adsorption, such as hydrogen bond, van der waals force, chemical bond, etc., and the physical adsorption is easier to wash away than the chemical adsorption. Hydroxyl exists on the surface of hydroxyapatite, can form hydrogen bonds with regorafenib molecules, can adsorb regorafenib by virtue of van der waals force, can form chemical bonds with regorafenib molecules by virtue of a surface modifier to be tightly combined, and loads regorafenib on the surface of hydroxyapatite, so that the permeability of regorafenib on tumor cell membranes is improved, and the regorafenib is brought into tumor cells. The drug is then released inside the cells, which can increase the intracellular drug concentration and decrease the effective drug concentration, which can reduce the damage to normal tissues.
(2) The nano-hydroxyapatite composite material prepared by the invention is prepared by fully mixing spherical porous hydroxyapatite and medicine regorafenib to prepare a suspension, preliminarily absorbing the medicine by utilizing the porosity of the hydroxyapatite, and further modifying free medicine and a medicine-containing hydroxyapatite preparation in a solution by utilizing the film forming property and the curling property of an amino acid monomer in the process of polymerizing to form a high polymer, thereby finally forming the nano sustained-release preparation taking the medicine-carrying porous hydroxyapatite as a core and the high polymer as a film coating. When the release medium exists, the drug loaded in the polymer film, the hydroxyapatite core and the surface can be slowly released at different time along with the swelling characteristic of the polymer. On one hand, the slow release microspheres improve the easy aggregation of inorganic carrier hydroxyapatite particles through the addition of organic polymers, and reduce the particle size of the hydroxyapatite particles, so that the hydroxyapatite particles reach the nanometer level and are easier to enter the interior of tumor cells; on the other hand, through modification, the release of the drug adsorbed on the surface of the hydroxyapatite particle is slowed down, so that the phenomenon of sudden release of the drug of the nano particles is avoided.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated. Unless otherwise specified, reagents and starting materials for use in the present invention are commercially available.
Example 1
A preparation method of a regorafenib coated nano-porous hydroxyapatite sustained release microsphere comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving Tween-80 in 0.5% toluene, adding 70ml mixed solution of toluene and Tween-80 into 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 11 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2.5h in the solution, separating and filtering the obtained product, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and drying in an oven at 80 ℃ for 10h to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of solution is dripped into the suspension at the speed of 2 ml/min; regulating pH value to 11 with 20% NaOH solution at normal pressure and 60 deg.C in thermostatic water bath, reacting for a certain time, separating out product, and adding distilled waterWashing until the solution is neutral, washing for 3 times by absolute ethyl alcohol, and drying in an oven at 80 ℃ for 10 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: weighing 10.0g of glutamic acid monomer, placing the glutamic acid monomer in a 250ml round bottom flask, adding 1.0ml of 85% phosphoric acid solution and 2.0ml of distilled water, fully and uniformly mixing, gradually heating the mixture to 200 ℃ under the condition of oil bath, simultaneously vacuumizing, reacting for 1.5h, cooling to 80 ℃ of oil bath temperature after the reaction is finished, and adding 30ml of 80% ethanol water solution to completely dissolve the product; and slowly dripping 10ml of the dissolved solution into 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature under the condition of stirring, filtering and drying the resultant to obtain the slow-release microspheres. The microsphere drug loading was determined to be 7.86%.
Example 2
A preparation method of a regorafenib coated nano-porous hydroxyapatite sustained release microsphere comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving Tween-80 in 0.5% toluene, adding 70ml mixed solution of toluene and Tween-80 into 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 11 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2.5h in the solution, separating and filtering the obtained product, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and drying in an oven at 80 ℃ for 10h to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of solution is dripped into the suspension at the speed of 2 ml/min; regulating the pH value to 11 by using a NaOH solution with the mass percent of 20% under the conditions of normal pressure and constant temperature water bath at 60 ℃, separating a product after reacting for a certain time, washing the product by using distilled water until the solution is neutral, washing the product for 3 times by using absolute ethyl alcohol, and drying the product in an oven at 80 ℃ for 10 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: weighing 10.0g of aspartic acid monomer, placing the aspartic acid monomer in a 250ml round bottom flask, adding 1.0ml of 85% phosphoric acid solution and 2.0ml of distilled water, fully and uniformly mixing, gradually heating the mixture to 200 ℃ under the condition of oil bath, simultaneously vacuumizing, reacting for 1.5h, cooling to 80 ℃ of oil bath temperature after the reaction is finished, and adding 30ml of 80% ethanol water solution to completely dissolve the product; and slowly dripping 10ml of the dissolved solution into 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature under the condition of stirring, filtering and drying the resultant to obtain the slow-release microspheres. The drug loading of the microspheres was determined to be 7.10%.
Example 3
A preparation method of a regorafenib coated nano-porous hydroxyapatite sustained release microsphere comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving Tween-80 in 0.5% toluene, adding 70ml mixed solution of toluene and Tween-80 into 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 11 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2.5h in the solution, separating and filtering the obtained product, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and drying in an oven at 80 ℃ for 10h to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of solution is dripped into the suspension at the speed of 2 ml/min; regulating the pH value to 11 by using a NaOH solution with the mass percent of 20% under the conditions of normal pressure and constant temperature water bath at 60 ℃, separating a product after reacting for a certain time, washing the product by using distilled water until the solution is neutral, washing the product for 3 times by using absolute ethyl alcohol, and drying the product in an oven at 80 ℃ for 10 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: weighing 10.0g of lysine monomer, placing the lysine monomer in a 250ml round bottom flask, adding 1.0ml of 85% phosphoric acid solution and 2.0ml of distilled water, fully and uniformly mixing, gradually heating the mixture to 200 ℃ under the condition of oil bath, simultaneously vacuumizing, reacting for 1.5h, cooling to 80 ℃ of oil bath temperature after the reaction is finished, and adding 30ml of 80% ethanol water solution to completely dissolve the product; and slowly dripping 10ml of the dissolved solution into 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature under the condition of stirring, filtering and drying the resultant to obtain the slow-release microspheres. The microsphere drug loading was determined to be 8.09%.
Comparative example 1
A preparation method of a regorafenib coated nano-porous hydroxyapatite sustained release microsphere comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving Tween-80 in 0.5% toluene, adding 70ml mixed solution of toluene and Tween-80 into 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 11 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2.5h in the solution, separating and filtering the obtained product, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and drying in an oven at 80 ℃ for 10h to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of solution is dripped into the suspension at the speed of 2 ml/min; regulating the pH value to 11 by using a NaOH solution with the mass percent of 20% under the conditions of normal pressure and constant temperature water bath at 60 ℃, separating a product after reacting for a certain time, washing the product by using distilled water until the solution is neutral, washing the product for 3 times by using absolute ethyl alcohol, and drying the product in an oven at 80 ℃ for 10 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: stirring 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature, filtering the resultant, and drying to obtain the slow-release microspheres. The microsphere drug loading was determined to be 6.69%.
Research on drug release performance of regorafenib-coated nano-porous hydroxyapatite sustained-release microspheres
Accurately transferring 20ml of Phosphate Buffer Solution (PBS) into a 50ml plastic centrifuge tube, precisely weighing 10mg of the sustained-release microspheres prepared in the examples 1-3 and the comparative example 1, placing the microspheres into the PBS solution, placing the reagent into a water bath constant-temperature shaking table for fixing, and adjusting the working condition of the constant-temperature shaking table to be 37.3 ℃. Another 50ml centrifuge tube 1 was transferred to PBS 20ml and put into a water bath constant temperature shaking table for use. Starting timing when the samples are put in, taking out the samples at 1, 2, 4, 6, 12, 24, 36, 48, 60, 72, 84, 96, 108 and 120h respectively, washing the samples with distilled water, putting the samples into standby centrifuge tubes, detecting samples taken out at different time intervals respectively by using an ultraviolet spectrophotometer, and calculating the cumulative release rate (%) of the drug regorafenib. The experiment was set up in 3 replicates. The results of the experiment are shown in table 1.
TABLE 1 in vitro cumulative release process of regorafenib
Figure BDA0002958813900000071
Figure BDA0002958813900000081
As can be seen from the experimental results in table 1, regorafenib is suddenly released from the surface of the nanoporous hydroxyapatite within 0-4h of the initial stage, and these may be physically adsorbed drug molecules, which are easily eluted because the physical adsorption is not tight; the release rate of the drug is reduced in different degrees within the range of 4-96h, and the cumulative release rates of the drugs of examples 1-3 exceed 90% within 96 h; in the range of 96-120h, the drug release level is basically balanced, and regorafenib cannot be completely released from the nano-porous hydroxyapatite, mainly because the nano-porous hydroxyapatite and regorafenib molecules have stronger adsorption. Specifically, the cumulative release rate of the polyglutamic acid modified sustained-release microspheres in example 1 reaches 92.7%, the cumulative release rate of the polyaspartic acid modified sustained-release microspheres in example 2 reaches 93.3%, and the cumulative release rate of the polylysine modified sustained-release microspheres in example 3 reaches 96.6%. Therefore, the polylysine modified sustained-release microspheres have the optimal accumulative release rate. In addition, compared with the slow release microspheres which are not modified by polyamino acid, the regorafenib coated nano porous hydroxyapatite slow release microspheres have more stable release rate, and can continuously and stably supply the drug active molecules within 120 h.
Tumor inhibition experiment of regorafenib-coated nano porous hydroxyapatite sustained release microspheres
90 colorectal cancer-bearing mice were randomly divided into 6 groups of 15 mice each. I, II groups of tumor-surrounding injection nano-porous hydroxyapatite sustained-release microspheres coated with regorafenib prepared in example 3, wherein the regorafenib doses are 200mg/kg and 100mg/kg respectively; in the III and IV groups, regorafenib injection is directly injected around tumor, and the doses of regorafenib are respectively 200mg/kg and 100 mg/kg; v group tumor is injected with nano porous hydroxyapatite sustained release microspheres with the dosage of 500 mg/kg; group VI was blank and was not treated. I. Groups II, III, IV and V were injected at 0d and 5d, respectively, and were injected around the tumor margin 5mm from the tumor margin. The microspheres are suspended and dissolved in iodized oil before injection, and the injection amount is 0.2 ml.
The survival status of the nude mice was observed at 0d, 5d, and 10d, and the tumor size was measured. Tumor volume calculation formula: v ═ B (axb)2) And/2, (A is the major diameter, B is the minor diameter). Mice were sacrificed at 10d, tumor weights were weighed, and tumor inhibition rates were calculated as (1-mean tumor weight in dosing group/mean tumor weight in control group) × 100%. The results of the experiment are shown in table 2.
TABLE 2 Effect of Regorafenib-coated nanoporous hydroxyapatite sustained release microspheres on tumor growth in colorectal cancer tumor-bearing mice
Figure BDA0002958813900000091
The experimental results in table 2 show that the injection of the regorafenib-coated nanoporous hydroxyapatite sustained release microspheres can continuously and significantly inhibit the growth of tumors of colorectal cancer tumor-bearing mice within 10 days, reduce the tumor volume and the tumor weight, show a positive dose correlation, and achieve a tumor inhibition rate of the drug-loaded sustained release microspheres with a dose of 200mg/kg of 69.7%. Compared with the existing injection, the injection of regorafenib has low tumor inhibition rate, which is directly related to the short tumor inhibition time and the rapid metabolism of the active ingredients of the medicine. The experimental results prove that the regorafenib-coated nano-porous hydroxyapatite sustained-release microsphere tumor-peripheral injection sustained-release chemotherapy has a reliable treatment effect on cancer transplantable tumors and has an important clinical application value.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications or equivalents may be made to the technical solution without departing from the principle of the present invention, and these modifications or equivalents should also be regarded as the protection scope of the present invention.

Claims (2)

1. The regorafenib-coated nano porous hydroxyapatite sustained release microsphere is characterized by comprising the following components in parts by weight: 5-10 parts of regorafenib and 10-20 parts of nano porous hydroxyapatite; the nano-porous hydroxyapatite is modified by polyamino acid,
wherein the polyamino acid is polyglutamic acid, polyaspartic acid or polylysine,
the preparation method of the regorafenib-coated nano-porous hydroxyapatite sustained-release microspheres comprises the following steps:
(1) preparation of porous spherical calcium carbonate: dissolving surfactant in 0.5% toluene, mixing 70ml toluene and surfactant solution in 30ml 1mol/L sodium carbonate solution, and ultrasonic emulsifying for 10-12 min; pouring the emulsion into CaCl with the concentration of 0.2mol/L2Stirring for 2-3 hr to obtain product, separating, filtering, repeatedly washing with distilled water, washing with anhydrous ethanol for 3 times, and oven drying at 80 deg.C for 8-12 hr to obtain porous spherical calcium carbonate powder;
(2) preparing porous hydroxyapatite: weighing 1g of porous spherical calcium carbonate powder, adding into 200ml of water to prepare suspension, and stirring 0.03mol/L of Na2HPO4200ml of the solution was added dropwise to the suspension at a rate of 2ml/min(ii) a Regulating the pH value to 10-12 by using a NaOH solution with the mass percent of 20% under the conditions of normal pressure and constant temperature water bath at 60 ℃, separating a product after reacting for a certain time, washing the product with distilled water until the solution is neutral, washing the product with absolute ethyl alcohol for 3 times, and drying the product in an oven at 80 ℃ for 8-12 hours to obtain porous hydroxyapatite;
(3) preparing a regorafenib coated nano porous hydroxyapatite sustained release microsphere: weighing 10.0g of amino acid monomer, placing the amino acid monomer in a 250ml round bottom flask, adding 1.0ml of 85% phosphoric acid solution and 2.0ml of distilled water, fully and uniformly mixing, gradually heating the mixture to 220 ℃ under the condition of oil bath, simultaneously vacuumizing, reacting for 1-2h, cooling to 80 ℃ of oil bath temperature after the reaction is finished, and adding 30ml of 80% ethanol water solution to completely dissolve the product; slowly dripping 10ml of the dissolved solution into 250ml of suspension containing 1.0g of regorafenib and 2.0g of porous hydroxyapatite at room temperature under the condition of stirring, filtering and drying the resultant to obtain the slow-release microspheres;
wherein the surfactant in the step (1) is tween-80 or lecithin,
the drying time of the drying oven in the steps (1) and (2) is 10h,
the oil bath heating temperature in the step (3) is 200 ℃.
2. Use of the regorafenib coated nanoporous hydroxyapatite sustained release microspheres according to claim 1 for the preparation of a medicament for the treatment of cancer, which is colorectal cancer or liver cancer.
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CN105169492A (en) * 2015-09-07 2015-12-23 广东省微生物研究所 Gamma-polyglutamic acid/hydroxyapatite gel microsphere carrier material and preparation method thereof
CN107233318A (en) * 2017-06-02 2017-10-10 河南工程学院 The preparation method of hydroxyapatite drug bearing microsphere with multistage slow controlled-release effect
CN111053751A (en) * 2018-10-16 2020-04-24 正大天晴药业集团股份有限公司 Regorafenib sustained-release tablet and preparation method thereof

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CN102153060A (en) * 2011-05-12 2011-08-17 合肥工业大学 Method for preparing hydroxyapatite nano-structure microspheres
CN105169492A (en) * 2015-09-07 2015-12-23 广东省微生物研究所 Gamma-polyglutamic acid/hydroxyapatite gel microsphere carrier material and preparation method thereof
CN107233318A (en) * 2017-06-02 2017-10-10 河南工程学院 The preparation method of hydroxyapatite drug bearing microsphere with multistage slow controlled-release effect
CN111053751A (en) * 2018-10-16 2020-04-24 正大天晴药业集团股份有限公司 Regorafenib sustained-release tablet and preparation method thereof

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