Developable embolic bead capable of loading radioactive iodine and negatively charged drug, preparation method therefor, and use thereof

Abstract

Disclosed are a developable embolic bead capable of loading radioactive iodine and a negatively charged drug, a preparation method therefor, and a use thereof. The bead is a quaternary ammonium salt bead containing a catechol structure and elemental iodine, and can be loaded with radioactive iodine and a negatively charged drug. In the present invention, a quaternary ammonium salt bead containing a catechol structure is prepared, using a polyamine polymer or a hydrochloride thereof, a catechol monomer, and an iodine-containing monomer as primary raw materials; the bead can be loaded with radioactive iodine and a negatively charged drug, thereby achieving combined radiotherapy, chemotherapy, and embolization treatment, as well as development diagnosis, of tumors.

Description

A visual embolization microsphere capable of loading radioactive iodine and negatively charged drugs, and a preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomedical materials, and particularly relates to a visual embolization microsphere capable of loading radioactive iodine and negatively charged drugs, and a preparation method and application thereof. BACKGROUND

[0002] Transcatheter arterial chemoembolization (TACE) is a common clinical interventional method for treating primary liver cancer, which is a method of performing intravascular chemotherapy and embolization on lesions in the liver by placing a catheter in the hepatic artery of the liver through a minimally invasive method. Drug-eluting beads (DEB) can have the effects of embolization and chemotherapy, and currently, there are several drug-eluting beads on the market, such as DC bead, Hepasphere, Callisphere, etc. These microspheres mainly use the negative charge of carboxylic acid or sulfonic acid groups to adsorb positively charged chemotherapeutic drugs to improve the treatment effect of embolization, but they cannot adsorb negatively charged drugs.

[0003] During chemotherapy, tumor cells often develop drug resistance, which reduces the treatment effect. In addition to killing tumor cells, radiotherapy can also reduce the drug resistance of tumor cells. Therefore, if chemotherapy and radiotherapy are combined, the treatment effect of embolization can be greatly improved. Therefore, it is of great significance to develop a drug-eluting microsphere capable of loading radioactive elements to reduce drug resistance in embolization therapy and improve the treatment effect of embolization. In addition, if the microsphere can be visualized during the operation or after the operation, the doctor can more accurately aim at the treatment area, detect and determine the endpoint, identify the area of insufficient treatment during the operation, improve the safety of the patient during the operation, and provide a basis for postoperative diagnosis. SUMMARY

[0004] The present application provides a visual embolization microsphere and a visual embolization microsphere capable of loading radioactive iodine and negatively charged drugs to provide a new reagent or drug for embolization, radiotherapy and chemotherapy for treating tumors in clinic, and significantly improve the treatment effect and reduce drug resistance.

[0005] The present application also provides a preparation method and application of the visual embolization microsphere capable of loading radioactive iodine and negatively charged drugs.

[0006] Technical scheme: In order to achieve the above-mentioned purpose, the present application provides a visual embolization microsphere, wherein the microsphere is a quaternary ammonium salt type microsphere containing catechol structure and iodine element.

[0007] The preparation method of the developed embolism microspheres comprises the following steps:

[0008] (1) Water phase preparation: dissolve polyamino polymer or its hydrochloride, catechol monomer, iodine-containing monomer in water;

[0009] (2) Oil phase preparation: add emulsifier to low-polarity solvent, and stir until dissolved;

[0010] (3) Microsphere preparation: add water phase to oil phase under stirring, add 3-halogen-1,2-epoxypropane, and after heating reaction, wash and filter.

[0011] In step (1), the polyamino polymer is one or more of polyallylamine and polyethyleneimine; the hydrochloride of the polyamino polymer is one or more of polyallylamine hydrochloride and polyethyleneimine hydrochloride; the catechol monomer is one or more of dopamine and dopamine hydrochloride; and the iodine-containing monomer is one or more of 5-amino-2,4,6-triiodoisophthalic acid, 3-acetylamino-5-amino-2,4,6-triiodobenzoic acid, 5-amino-N-(2-hydroxyethyl)-2,4,6-triiodoisophthalic acid, and 3-amino-2,4,6-triiodobenzoic acid.

[0012] In step (1), 100 parts of polyamino polymer or its hydrochloride, 1-40 parts of catechol monomer, and 10-200 parts of iodine-containing monomer are dissolved in 100-1000 parts of pure water by weight; when polyamino polymer or its hydrochloride is used, 5-60 parts of an alkaline compound needs to be added, and the alkaline compound is one or more of sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, pyridine, triethylamine, sodium acetate, sodium hydroxide, and potassium hydroxide.

[0013] In step (2), the low-polarity solvent includes one or more of liquid paraffin, diethyl ether, petroleum ether, hexane, cyclohexane, and heptane; the emulsifier is one or more of Span, Tween, lauryl alcohol polyoxyethylene (120) ether, polyethylene glycol (200) monolaurate, glycerol monolaurate, N,N-dimethylhexanamide, polyglycerol fatty acid ester, polyethylene glycol (200) dilaurate, diglycerol monolaurate, sorbitol monolaurate, and nonyl phenoxypolyethoxy ethanol; 0.5-5 parts of emulsifier is added in 2-10 times the volume of the low-polarity solvent of the water phase, and the emulsifier is stirred until dissolved.

[0014] Wherein, in step (3), the water phase is added to the oil phase under mechanical stirring of 100-500 rpm, 10-50 parts of 3-halogenated-1,2-epoxypropane is added, wherein the amount of 3-halogenated-1,2-epoxypropane is added according to the weight ratio of each substance in the water phase, the temperature is raised to 30-100℃, and the reaction is carried out for 2-24h, and then washing and filtering are carried out to obtain the product; the 3-halogenated-1,2-epoxypropane is one or more of epoxy chloropropane and epoxy bromopropane.

[0015] The imaging embolization microspheres loaded with radioactive iodine and negative charge drugs according to the present application are quaternary ammonium salt type microspheres containing catechol structure and iodine element, and loaded with radioactive iodine and negative charge drugs.

[0016] The preparation of the imaging embolization microspheres loaded with radioactive iodine and negative charge drugs according to the present application comprises the following steps:

[0017] (1) Loading radioactive iodine: the prepared microspheres are placed in a container containing a catalyst, and then a radioactive iodide solution is added, and after centrifugation, the supernatant is discarded to obtain microspheres loaded with radioactive iodine;

[0018] (2) Loading negative charge radiotherapy / chemotherapy drugs: the prepared microspheres loaded with radioactive iodine are placed in a solution containing negative charge drugs, and are shaken and stirred, and then centrifuged and the supernatant is discarded to obtain microspheres loaded with negative charge radiotherapy / chemotherapy drugs; for the microspheres prepared by using polyamino polymers, acidification needs to be carried out before loading the negative charge radiotherapy / chemotherapy drugs: the microspheres are placed in an acidic compound solution, and then centrifuged and the supernatant is discarded.

[0019] The acidic compound is one or more of hydrochloric acid, sulfuric acid, acetic acid and phosphoric acid.

[0020] The radioactive iodide is one or more of

[0125] sodium iodide and

[0131] sodium iodide; the catalyst is one or more of chloramine-T and 1,3,4,6-tetrachloro-3α,6α-diphenylglycoluril; and the negative charge drug is one or more of carboxylate, phosphate, phosphate, nucleotide, deoxynucleotide, cyclic dinucleotide drug, oncolytic virus and

[0032] Na2HPO4.

[0021] As a preference, the negative charge drug is one or more of combretastin phosphate disodium (CA4P), combretastin A1 disodium phosphate tetrasodium salt (OXi-4503sodium, CA1P), STING agonist ADU-S100, MDK-563729, E7766, MK-1454, MSA-2, TAK-676, SR-717, sodium norcantharidin, mRNA-2416, oncolytic adenovirus,

[0032] Na2HPO4,

[0125] NaI and

[0131] NaI.

[0022] The application of the radioiodine and negative charge drug loaded imaging embolization microspheres in the preparation of tumor radiotherapy, chemotherapy and embolization combined treatment drugs and imaging diagnostic reagents

[0023] The microspheres can be prepared by adjusting the amount of emulsifier, stirring speed and oil-water ratio according to the technical report.

[0024] The quaternary ammonium salt type microspheres containing catechol structure are prepared by using polyamino polymer or hydrochloride thereof, catechol monomer and iodine-containing monomer as main raw materials, and can load radioiodine and negative charge drugs to realize tumor radiotherapy, chemotherapy and embolization combined treatment and imaging diagnosis.

[0025] The quaternary ammonium salt type microspheres containing catechol structure are prepared by using polyamino polymer or hydrochloride thereof, catechol monomer and iodine-containing monomer as main raw materials, and can load radioiodine and negative charge drugs to realize tumor radiotherapy, chemotherapy and embolization combined treatment and imaging diagnosis.

[0026] The quaternary ammonium salt type microspheres containing catechol structure are prepared by using polyamino polymer or hydrochloride thereof, catechol monomer and iodine-containing monomer as main raw materials, and can load radioiodine and negative charge drugs to realize tumor radiotherapy, chemotherapy and embolization combined treatment and imaging diagnosis.

[0027] Advantages: compared with the prior art, the present application has the following advantages:

[0028] 1. The quaternary ammonium salt type microspheres containing catechol structure are prepared by using polyamino polymer or hydrochloride thereof, catechol monomer and iodine-containing monomer as main raw materials, and can load radioiodine and negative charge drugs to realize tumor radiotherapy, chemotherapy and embolization combined treatment and imaging diagnosis.

[0029] 2、The microspheres prepared by the application can also load radioactive iodine elements and negative charged radiotherapy and chemotherapy drugs, and one microsphere can load multiple radioactive iodine elements and multiple negative charged drugs, realizing combined treatment of radiotherapy, chemotherapy and embolism, and can significantly improve the treatment effect of embolism.

[0030] 3、The microspheres prepared by the application can be developed during the operation process or after the operation, doctors can more accurately aim at the treatment area, detect and determine the end point, identify the area of insufficient treatment during the operation process, improve the safety of patients during the operation process, and provide the basis for postoperative diagnosis. BRIEF DESCRIPTION OF DRAWINGS

[0031] Fig. 1 is an SEM spectrum of the microspheres prepared in Example 1 of the application;

[0032] Fig. 2 is an infrared spectrum of the microspheres prepared in Example 1 of the application, and the spectrum shows that there are characteristic peaks of quaternary ammonium salt at 2000 cm -1 and 3000 cm -1 , which indicates that the microspheres are successfully synthesized.

[0033] Fig. 3 is the development of the microspheres under DSA, A-D are respectively the microspheres prepared in Example 8, Example 7, Example 6 and Example 1.

[0034] Fig. 4 is a bar chart of the tumor size volume ratio after treatment and difference analysis, "*" represents p≤0.05, "**" represents p≤0.01, "***" represents p≤0.001, and "****" represents p≤0.0001 DETAILED DESCRIPTION

[0035] The application will be further described below in combination with examples and drawings.

[0036] The materials, reagents and the like used in the following examples can be obtained from commercial channels unless otherwise specified. The experimental methods not specified in the examples are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer.

[0037] The raw materials in the embodiments of the present application, such as polyallylamine hydrochloride (polyallylamine hydrochloride, polyallylamine hydrochloride, CAS: 71550-12-4, average Mw = 17500, Merck, item number 283215), polyethyleneimine (Aldrich, item number E107079, average Mw = 10000), polyallylamine (Aldrich, item number P432586, average Mw = 15000); 5-amino-2,4,6-triiodoisophthalic acid, dopamine hydrochloride (dopamine hydrochloride), liquid paraffin, span 80, epichlorohydrin, 5-amino-N-(2-hydroxyethyl)-2,4,6-triiodoisophthalic acid, 3-amino-2,4,6-triiodobenzoic acid, 5-amino-2,4,6-triiodoisophthalic acid,

[0131] NaI (Na 131 I solution, Atom High-Tech Co., Ltd.), CA1P, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril, etc. are commercially available and can be purchased from reagent companies such as Merck, Aldrich, and McKinley, or similar reagents from pharmaceutical companies.

[0038] Example 1

[0039] Preparation of microspheres

[0040] Add 25 mL of liquid paraffin and 10 μL of span 80 to a 50 mL two-necked flask with mechanical stirring, adjust the stirring speed to 150 rpm, and stir for 0.5 h to obtain an oil phase. Then, 1600 mg of polyallylamine hydrochloride, 1600 mg of 5-amino-2,4,6-triiodoisophthalic acid, 200 mg of dopamine hydrochloride, and 300 mg of NaOH are dissolved in 3 mL of pure water to obtain an aqueous phase, which is added dropwise to the oil phase flask, and stirred at 150 rpm for 1 h. Then, 200 μL of epichlorohydrin is added, and the reaction is carried out at 50°C for 24 h. After the reaction is completed, the product is washed with 250 mL of isopropyl alcohol and 100 mL of ethanol for 1 h, respectively, and then suction filtered. Finally, the filter residue is washed with a hydrochloric acid solution, filtered, and dried. The infrared spectrum of the microspheres prepared in this example is shown in FIG. 2, and the spectrum shows characteristic peaks of quaternary ammonium salt at 2000 cm -1 and 3000 cm -1 , indicating that the microspheres are successfully synthesized. The SEM image of the microspheres is shown in FIG. 1, and the prepared microspheres have regular morphology and good sphericity. The development effect of the microspheres is shown in FIG. 3D, and the development effect is good. DSA development: the microspheres are placed in a centrifuge tube, and physiological saline is added to swell for 1 h, and then centrifuged to remove the supernatant. The centrifuge tube with the microspheres is placed under DSA for contrast, and the development of the microspheres is observed. After reducing the amount of iodine-containing monomer, the development effect of the microspheres is significantly reduced.

[0041] Example 2

[0042] Preparation of microspheres

[0043] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 10 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1600 mg polyallylamine hydrochloride, 600 mg dopamine hydrochloride, 1600 mg 3-acetylamino-5-amino-2,4,6-triiodo-benzoic acid (cas: 1713-07-1), 400 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the above oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 220 μL epichlorohydrin was added, and reaction was performed at 50°C for 24 h. After the reaction was completed, washing was performed with 250 mL isopropyl alcohol, 100 mL ethanol, and suction filtration was performed for 1 h, and finally the filter residue was washed with a hydrochloric acid solution, filtered, and dried.

[0044] Example 3

[0045] Microsphere preparation

[0046] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 10 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1600 mg polyallylamine hydrochloride, 600 mg dopamine hydrochloride, 1600 mg 3-acetylamino-5-amino-2,4,6-triiodo-benzoic acid (cas: 1713-07-1), 400 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the above oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 220 μL epichlorohydrin was added, and reaction was performed at 50°C for 24 h. After the reaction was completed, washing was performed with 250 mL isopropyl alcohol, 100 mL ethanol, and suction filtration was performed for 1 h, and finally the filter residue was washed with a hydrochloric acid solution, filtered, and dried.

[0047] Example 4

[0048] Microsphere preparation

[0049] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 10 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1600 mg polyallylamine hydrochloride, 600 mg dopamine hydrochloride, 1600 mg 3-acetylamino-5-amino-2,4,6-triiodo-benzoic acid (cas: 1713-07-1), 400 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the above oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 220 μL epichlorohydrin was added, and reaction was performed at 50°C for 24 h. After the reaction was completed, washing was performed with 250 mL isopropyl alcohol, 100 mL ethanol, and suction filtration was performed for 1 h, and finally the filter residue was washed with a hydrochloric acid solution, filtered, and dried.

[0050] Example 5

[0051] Microsphere preparation

[0052] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 15 μL span 80 were added, the stirring speed was adjusted to 140 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1800 mg polyallylamine hydrochloride, 180 mg dopamine, 1100 mg 5-amino-2,4,6-triiodoisophthalic acid, and 250 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the oil phase flask, and stirring was performed at 140 rpm for 1 h. Then 250 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. After the reaction was completed, washing was performed with 250 mL isopropyl alcohol and 50 mL ethanol for 1 h, and suction filtration was performed, and finally the filter residue was washed with a sulfuric acid solution, filtered, and dried.

[0053] Example 6

[0054] Microsphere preparation

[0055] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 20 μL span 80 were added, the stirring speed was adjusted to 110 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1600 mg polyallylamine hydrochloride, 390 mg dopamine, 800 mg 5-amino-2,4,6-triiodoisophthalic acid, and 400 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the flask, and stirring was performed at 110 rpm for 1 h. Then 220 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. After the reaction was completed, washing was performed with 220 mL isopropyl alcohol and 150 mL ethanol for 1 h, and suction filtration was performed, and finally the filter residue was washed with a hydrochloric acid solution, filtered, and dried. The prepared microspheres were developed by DSA as shown in FIG. 3B.

[0056] Example 7

[0057] Microsphere preparation

[0058] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 15 μL span 80 were added, the stirring speed was adjusted to 160 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 2500 mg polyethyleneimine, 300 mg 5-amino-2,4,6-triiodoisophthalic acid, 390 mg dopamine, and 100 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the oil phase flask, and stirring was performed at 160 rpm for 1 h. Then 210 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. After the reaction was completed, washing was performed with 250 mL isopropyl alcohol and 100 mL methanol for 1 h, and suction filtration was performed, and finally the filter residue was washed with a hydrochloric acid solution, filtered, and dried. The prepared microspheres were developed by DSA as shown in FIG. 3C.

[0059] Example 8

[0060] Microsphere preparation

[0061] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 25 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h to obtain an oil phase. Then 1000 mg polyallylamine, 200 mg dopamine, 400 mg 5-amino-2,4,6-triiodoisophthalic acid, and 270 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 200 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. After the reaction was completed, the solution was washed with 200 mL petroleum ether and 100 mL ethanol for 1 h, and suction filtration was performed. Finally, the filter residue was washed with a sulfuric acid solution, filtered, and dried. The prepared microspheres DSA development is shown in FIG. 3A.

[0062] Comparative Example 1

[0063] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 10 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h. Then 1600 mg polyallylamine hydrochloride, 400 mg dopamine hydrochloride, 1600 mg 5-amino-2,4,6-triiodoisophthalic acid, and 800 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 200 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. After the reaction was completed, the solution was washed with 250 mL isopropanol and 100 mL ethanol for 1 h, and suction filtration was performed. Finally, the filter residue was washed with a hydrochloric acid solution, filtered, and dried. Due to the increased amount of NaOH added, a large amount of NaCl was generated during the reaction, the emulsion was broken during the reaction, and the microspheres contained a large number of fragments.

[0064] Comparative Example 2

[0065] Into a 50 mL two-necked flask with mechanical stirring, 25 mL liquid paraffin and 10 μL span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h. Then 1600 mg polyallylamine hydrochloride, 1600 mg 5-amino-2,4,6-triiodoisophthalic acid, 1000 mg dopamine hydrochloride, and 300 mg NaOH were dissolved in 3 mL pure water to obtain an aqueous phase, which was added dropwise into the oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 200 μL epichlorohydrin was added, and reaction was performed at 50 °C for 24 h. A large amount of dopamine hydrochloride remained undissolved in the solution after the reaction was completed.

[0066] Comparative Example 3

[0067] Into a 50 mL two-necked flask with mechanical stirring, 25 mL of liquid paraffin and 10 μL of span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h. Then 1600 mg of polyallylamine hydrochloride, 90 mg of dopamine hydrochloride, 1600 mg of 5-amino-2,4,6-triiodoisophthalic acid, and 300 mg of NaOH were dissolved in 3 mL of pure water, and the solution was added dropwise to the above oil phase flask, and stirring was performed at 150 rpm for 1 h. Then 200 μL of epichlorohydrin was added, and the reaction was performed at 50°C for 24 h. After the reaction was completed, 250 mL of isopropyl alcohol and 100 mL of ethanol were used for washing for 1 h and suction filtration, respectively, and finally, the product was washed with a hydrochloric acid solution, filtered, and dried.

[0068] Comparative Example 4

[0069] Into a 50 mL two-necked flask with mechanical stirring, 25 mL of liquid paraffin and 20 μL of span 80 were added, the stirring speed was adjusted to 170 rpm, and stirring was performed for 0.5 h. Then 1600 mg of gelatin, 200 mg of dopamine hydrochloride, 1600 mg of 5-amino-2,4,6-triiodoisophthalic acid, and 300 mg of NaOH were dissolved in 3 mL of pure water, and the solution was added dropwise to the above oil phase flask, and stirring was performed for 1 h. Then 200 μL of epichlorohydrin was added, and the reaction was performed at 50°C for 24 h. After the reaction was completed, 250 mL of isopropyl alcohol and 100 mL of ethanol were used for washing for 1 h and suction filtration, respectively, and finally, the product was washed with a hydrochloric acid solution, filtered, and dried.

[0070] Comparative Example 5

[0071] Into a 50 mL two-necked flask with mechanical stirring, 25 mL of liquid paraffin and 10 μL of span 80 were added, the stirring speed was adjusted to 150 rpm, and stirring was performed for 0.5 h. Then 1600 mg of polyallylamine hydrochloride, 2400 mg of 5-amino-2,4,6-triiodoisophthalic acid, 200 mg of dopamine hydrochloride, and 500 mg of NaOH were dissolved in 3 mL of pure water, and the solution was added dropwise to the above oil phase flask, and stirring was performed for 1 h. Then 200 μL of epichlorohydrin was added, and the reaction was performed at 50°C for 24 h. After the reaction was completed, 250 mL of isopropyl alcohol and 100 mL of ethanol were used for washing for 1 h and suction filtration, respectively, and finally, the product was washed with a hydrochloric acid solution, filtered, and dried. Since the amount of catechol monomer was increased, the amount of NaOH added was increased, and a large amount of NaCl was generated during the reaction, and the emulsion was unstable during the reaction, and the prepared microspheres were all broken into pieces.

[0072] Example 9

[0073] Study on the drug loading capacity of different microspheres

[0074] The drug-eluting microspheres prepared by different embodiments of the present application, Comparative Examples 3 and 4, are loaded with CA1P and radionuclides, and the drug addition amount is 0.2 mg / μL of microspheres and 2.0 μci / μL of microspheres, respectively. The loading method and the method for determining the drug loading amount are as follows:

[0075] (1) Complintrin A1 diphosphate tetrasodium salt (CA1P): An 80 ug / mL CA1P physiological saline solution is prepared, and after being diluted by 2, 4, 8, 16, and 32 times, respectively, the absorbance is determined at 295 nm by using an ultraviolet spectrophotometer, and a standard curve is plotted with the concentration as the ordinate and the absorbance as the abscissa. The microspheres are placed in a CA1P physiological saline solution (2.5 mg / mL) and shaken for 1 h, and then the supernatant is taken and the drug loading amount is calculated.

[0076] (2) Radioactive iodine (

[0131] NaI): The microspheres are placed in a 100 ul container containing 1, 3, 4, 6-tetrachloro-3α, 6α-diphenylglycoluril (1 mg / mL DMSO solution), and then a radioactive iodine solution (Na 131 I solution) is added, and after being shaken for 1 h, the radioactive count of the supernatant is determined, and the loading amount is calculated.

[0077] And the maximum drug loading amount (CA1P) is calculated according to the following formula: 上清 总 Maximum drug loading amount (

[0131] NaI) = (1 - M 上清 / M 总 ) × 2.0

[0078] In the formula, M 上清 is the total mass of the drug in the supernatant; M 总 is the total mass of the added drug; and if the drug is a radionuclide, M represents the radioactive count.

[0079] The drug loading amount of the drug-eluting microspheres prepared by different embodiments of the present application for CA1P and radionuclides is shown in Table 1, and the drug-eluting microspheres prepared in Example 1 have the best performance for loading a negatively charged drug and radioactive iodine. In Comparative Example 3, the amount of dopamine added during preparation is small, so the loading of radioactive iodine is low. In other examples, the microspheres have good performance for loading radioactive iodine and negatively charged drugs. In Comparative Example 4, gelatin and dopamine are copolymerized, and the microspheres prepared thereby have low efficiency for loading radionuclides and negatively charged drugs.

[0080] Table 1 Drug loading amount of microspheres prepared by different embodiments and comparative examples for CA1P and radioactive iodine

[0081] Example 10

[0082] Test of the ability of microspheres to load different types of iodine and drugs​

[0083] The microspheres prepared in Example 1 were loaded with different types of drugs, and the amount of drug added was 0.2 mg / μL of microspheres and 2.0 μCi / μL of microspheres, respectively. The loading and determination of drug loading amount were as follows:

[0084] (1) CA1P: according to the method (1) of Example 9.

[0085] (2) Sodium cantharidate: 1.0, 2.0, 3.0, 5.0, 7.0, and 10.0 ml of sodium cantharidate aqueous solution (1 mg / mL) were accurately measured into a 50 ml volumetric flask, respectively, and diluted to the mark with the mobile phase, and shaken well. 20 μL was injected into the chromatograph, respectively, and the chromatogram was recorded. The concentration (C, μg / mL) was taken as the abscissa, and the peak area (A) was taken as the ordinate, and linear regression was performed (chromatographic conditions: SHISEIDO TYPE MG ODS column (150 mm 4.6 mm, 5 μm), column temperature 30 ℃. 0.1 mol / L potassium phosphate dibasic-acetonitrile (80:20) (phosphoric acid was added to adjust the pH value to 2.6) was used as the mobile phase, and the detection wavelength was 210 nm). The microspheres were placed in the sodium cantharidate aqueous solution (1 mg / mL), shaken for 1 h, and the supernatant was tested by liquid chromatography and the drug loading amount was calculated.

[0086] (3) ADU-S100 (interferon gene stimulator (STING)): 0.01, 0.03, 0.06, and 0.12 mg / mL of ADU-S100 saline solution were prepared, respectively, 20 μL was injected into the chromatograph, and the chromatogram was recorded. The concentration (C, μg / mL) was taken as the abscissa, and the peak area (A) was taken as the ordinate, and linear regression was performed (chromatographic conditions: iPrOH: hexane = 10:90 as the mobile phase. The column temperature was 25 ℃. The flow rate was 1.0 mL / min, and the detection wavelength was 210 nm). The microspheres were placed in the ADU-S100 saline solution (0.1 mg / mL), shaken for 1 h, and the supernatant was tested by liquid chromatography and the drug loading amount was calculated.

[0087] (4) mRNA-2416: mRNA-2416 saline solution of 0.1 mg / mL was prepared, microspheres were added, shaken for 1 h, and the supernatant was tested according to the method of GB / T 34796-2017 to determine the content of mRNA-2416 in the supernatant, and the loading amount was calculated.

[0088] (5)

[0032] Na2HPO4: The microspheres were placed in the

[0032] Na2HPO4 solution, shaken for 0.5 h, and the radioactivity count of the supernatant was measured and the loading amount was calculated.

[0089] (6) Radioactive iodine: according to the method (2) of Example 9.

[0090] The drug loading amount of the microspheres is calculated by the following formula: Drug loading amount = (1 - M 上清 / M 总 ) x 100%

[0091] In the formula, M 上清 is the total mass of the drug in the supernatant; M 总 is the total mass of the added drug; and if the drug is a radionuclide, M represents the radioactivity count.

[0092] The drug loading amount of the microspheres prepared in Example 1 is shown in Table 2.

[0093] Table 2 Drug loading amount of the microspheres loaded with radiotherapy or chemotherapy drugs

[0094] As shown in Table 2, the microspheres prepared in Example 1 have good loading performance for various drugs.

[0095] Example 11

[0096] Study on the treatment effect of free drugs and microspheres loaded with different drugs on tumors

[0097] The microspheres prepared in Example 1 are loaded with different drugs to study the treatment effect on tumors.

[0098] The method for loading a single drug is according to the method in Examples 9-10.

[0099] The method for loading two drugs is as follows: the microspheres are placed in a container containing 100 μL of 1,3,4,6-tetrachloro-3α,6α-diphenylglycoluril (1 mg / mL DMSO solution), and then a solution of radioactive iodine (

[0131] NaI) (88.9 μci / μL microspheres) is added. After oscillation for 1 h, the supernatant is discarded after centrifugation, and the microspheres are dried. Then, a solution of CA1P (0.89 mg / μL microspheres) is added, oscillation is performed for 1 h, the supernatant is discarded after centrifugation, and the microspheres are dried.

[0100] Modeling: VX-II cells are used to build a tumor rabbit model in the muscle structure of healthy New Zealand rabbits. After the subcutaneous tumor diameter grows to 2 cm, the tumor is removed, and the tumor is cut into a cube with a side length of 3 mm for use. The healthy New Zealand rabbit is anesthetized, the abdominal cavity is opened, the left outer lobe of the liver is taken, and the liver is cut open. The tumor is embedded in the liver tissue, and the wound is blocked and sutured.

[0101] Drug administration: The specific drug administration amount is shown in Table 3. The drug administration amount of the microspheres loaded with CA1P +

[0131] NaI is half of the administration amount of free drugs CA1P and

[0131] NaI injection, and half of the administration amount of the microspheres loaded with CA1P alone and the microspheres loaded with

[0131] NaI alone.

[0102] The drug-loaded microspheres are administered as follows: DSA-guided VX-2 rabbit model interventional embolization - about 10-14 days after operation, enhanced CT is used to observe the volume of the model rabbit left liver lesion, and the interventional treatment is started when the lesion size is about 1 cm x 1 cm. After the animal model is anesthetized, the muscle layer of the right femoral artery of the rabbit is incised, and the femoral artery is stripped. The distal end is ligated, a 4F sheath is inserted through vascular puncture, and a 2.7F microcatheter is inserted into the femoral artery. Under the guidance of DSA, iodixanol enhanced angiography is performed, and the microcatheter is selected to the hepatic artery branch in front of the tumor lesion. Finally, the drug-loaded microspheres prepared in Example 1 are used for embolization, and finally the catheter, guide wire and sheath tube are removed, the proximal blood vessel is ligated, and the operation is completed. The free group is administered by catheter perfusion as in the microsphere group.

[0103] Tumor volume measurement: at 0, 7 days of treatment, CT scanning was performed to measure the length, width and height of the tumor, and the tumor volume was calculated using the following formula:

[0104] wherein a, b and c are the length, width and height of the tumor, respectively.

[0105] As shown in Table 3 and Figure 4, the treatment effect of the present application using low-dose co-loaded radionuclide and chemotherapeutic drug imaging embolization microspheres after interventional administration is significantly better than that of high-dose single chemotherapeutic embolization microspheres, single radiotherapy embolization microspheres and free drug radiotherapy and chemotherapy combined systemic administration, and there is a significant difference.

[0106] Table 3 Tumor size after treatment of New Zealand white rabbits VX II liver cancer model with different drug-loaded microspheres

Claims

1. A visualizing embolizing microsphere, characterized by, The microspheres are quaternary ammonium salt type microspheres containing catechol structure and iodine element.

2. A method of producing the visualizing embolizing microspheres according to claim 1, characterized by, The method comprises the following steps: (1) water phase preparation: dissolving polyamino polymer or its hydrochloride, catechol monomer and iodine-containing monomer in water; (2) oil phase preparation: adding emulsifier to low-polarity solvent and stirring until dissolved; (3) microsphere preparation: adding water phase to oil phase under stirring, adding 3-halo-1,2-epoxypropane, washing and filtering after heating reaction.

3. The method of claim 2, wherein the microspheres are formed by a process comprising: The polyamino polymer in step (1) is one or more of polyallylamine and polyethyleneimine; the hydrochloride of the polyamino polymer is one or more of polyallylamine hydrochloride and polyethyleneimine hydrochloride; the catechol monomer is one or more of dopamine and dopamine hydrochloride; and the iodine-containing monomer is one or more of 5-amino-2,4,6-triiodoisophthalic acid, 3-acetylamino-5-amino-2,4,6-triiodobenzoic acid, 5-amino-N-(2-hydroxyethyl)-2,4,6-triiodoisophthalic acid and 3-amino-2,4,6-triiodobenzoic acid.

4. The method of claim 2, wherein the microspheres are visualized by a method selected from the group consisting of radiography, computed tomography, magnetic resonance imaging, ultrasound imaging, and nuclear medicine imaging. In step (1), 100 parts of polyamino polymer or its hydrochloride, 1-40 parts of catechol monomer and 10-200 parts of iodine-containing monomer are dissolved in 100-1000 parts of pure water by weight; when polyamino polymer hydrochloride is used, 5-60 parts of alkaline compound is further added, and the alkaline compound is one or more of sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, pyridine, triethylamine, sodium acetate, sodium hydroxide and potassium hydroxide.

5. The method for preparing the imaging embolic microspheres according to claim 2, characterized in that, The low-polarity solvent in step (2) preferably comprises one or more of liquid paraffin, diethyl ether, petroleum ether, hexane, cyclohexane and heptane; and the emulsifier is one or more of Span, Tween, lauryl alcohol polyoxyethylene (120) ether, polyethylene glycol (200) monolaurate, glycerol monolaurate, N,N-dimethylhexanamide, polyglycerol fatty acid ester, polyethylene glycol (200) dilaurate, diglycerol monolaurate, sorbitan monolaurate and nonyl phenoxypolyethoxyethanol; wherein 0.5-5 parts of emulsifier is added in 2-10 times volume of low-polarity solvent of the water phase, and stirring is performed until dissolved.

6. The method of claim 2, wherein the microspheres are formed by a process comprising: In step (3), the water phase is added to the oil phase under mechanical stirring at 100-500 rpm, 10-50 parts of 3-halo-1,2-epoxypropane is added, the temperature is raised to 30-100℃, and reaction is performed for 2-24 h, followed by washing and filtering to obtain the product; and the 3-halo-1,2-epoxypropane is one or more of epichlorohydrin and epibromohydrin.

7. A radiopaque iodine and negatively charged drug loaded microsphere embolization microsphere characterized in that, The microspheres are quaternary ammonium salt type microspheres containing catechol structure and iodine element and loaded with radioactive iodine and negatively charged drugs.

8. A method for preparing radioiodine and negatively charged drug loaded visualization embolization microspheres, characterized by The method comprises the following steps: (1) loading radioactive iodine: placing the microspheres prepared in claim 2 in a container containing a catalyst, then adding a radioactive iodide solution, centrifuging and discarding the supernatant to obtain radioactive iodine-loaded microspheres; (2) loading negative charged drug: the microspheres loaded with radioactive iodine obtained in step (2) are placed in a solution containing a negative charged drug, and shaken and stirred, and then centrifuged to discard the supernatant to obtain microspheres loaded with radioactive iodine and the negative charged drug; for the microspheres prepared by using a polyamino polymer, the microspheres need to be acidified before loading the negative charged drug: the microspheres are placed in an acid compound solution, and then centrifuged to discard the supernatant.

9. The method for preparing radioactive iodine-loaded and negatively charged drug-loaded contrast-enhancing embolic microspheres according to claim 8, characterized in that, The radioactive iodide is one or more of [125] sodium iodide and [131] sodium iodide; the catalyst is one or more of chloramine-T and 1,3,4,6-tetrachloro-3α,6α-diphenylglycoluril; and the negative charged drug is one or more of carboxylate, phosphate, phosphate, nucleotide, deoxynucleotide, cyclic dinucleotide drug, oncolytic virus and [32] Na2HPO4.

10. Use of the microspheres loaded with radioactive iodine and negative charged drug according to claim 1 in the preparation of a combined radiotherapy, chemotherapy and embolization treatment drug for tumors and a visualized diagnostic reagent.

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