Multifunctional microsphere formulations and methods for preparing them for radiochemoembolization therapy and imaging of tumors.

Abstract

The present invention provides a microsphere formulation and a method for preparing it that can be used for tumor embolization therapy combined with radiotherapy and chemotherapy and nuclear medicine imaging. The microspheres are made of a polyvinyl alcohol derivative as a backbone material, and are polymerized and crosslinked with N-acryloyl amino acid monomers to harden them. They can be labeled with radioactive iodine and can also adsorb and carry chemotherapeutic agents.

Description

[Technical Field]

[0001] This invention belongs to the field of drug formulation, and specifically relates to tumors. chemistry Radiation therapy Embolization therapy and Amazing for This invention relates to a multifunctional microsphere formulation and a method for preparing the same. [Background technology]

[0002] Existing technologies show a trend of increasing malignant tumor incidence year after year. Correlative statistics show that the number of deaths from malignant tumors is the second leading cause of death worldwide, after cardiovascular disease. Currently, malignant Among tumor treatments, surgical resection, radiation therapy, and chemotherapy are the mainstays. Research has shown that most malignant tumors are rich in vascular systems, and these systems not only provide the nutrients necessary for tumor growth but also play a crucial role in late-stage metastasis. Tumor embolization therapy cuts off the supply of nutrients to the tumor by blocking tumor-specific blood supply vessels, effectively "starving" the tumor. Let This refers to the act of attempting to embolize. of When performing this procedure, the therapeutic effect can be enhanced by using chemotherapy agents or radioactive radionuclides in combination.

[0003] The embolic agents commonly used in chemotherapy-embolic treatment for tumors are Lipiodol and DC Bead. TM The chemotherapeutic agent aqueous solution and Lipiodol were pushed back against each other to form a stable W / O emulsion, which was then used for chemoembolization therapy for liver cancer. case, It can extend the patient's survival time to some extent. Correlation did By practice Then Furthermore, the time it takes for Lipiodol to embolize tumor blood vessels is short, and the chemotherapeutic agents in Lipiodol of rapid Na release Out This is a serious problem, as it is known to easily trigger toxic reactions in normal tissue. DC Bead TM teeth 、 As a solid embolic microsphere, Lipiodol ofDC Bead overcame many of the shortcomings of chemotherapy-embolization therapy. TM This method can form a permanent embolism, extending the embolism time, and also allows for the gradual release of the drug contained within it, thereby maintaining a high drug concentration at the tumor site while simultaneously lowering the drug concentration in the surrounding blood, thus avoiding systemic toxicity.

[0004] therapeutic radionuclides 131 I is an ideal internal irradiation radionuclide with a half-life of 8 days, emitting beta rays, a maximum radiation energy of 0.81 MeV, and a maximum penetration depth of 2 mm. However, it also emits gamma rays with a maximum energy of 364 keV, making it suitable for use in emission computed tomography or single-photon emission computed tomography. 131 I-labeled lipiodol has been used clinically in internal radiation embolization therapy for liver cancer, achieving a certain level of therapeutic effect. However, because lipiodol deposits in the lungs and is prone to causing side effects such as lung damage, its clinical application is limited. Currently, 131 Lipiodol products bearing the I label have been withdrawn by European producers.

[0005] Both chemotherapy-embolization and radiation-embolization can extend a patient's survival time to some extent, but there are limits to their therapeutic effects. therapy While some clinical trials have shown that combining external beam radiation therapy can further extend patient survival, the radiation damage to non-tumor areas caused by external beam radiation has significant side effects on patients, and therefore embolization therapy and chemotherapy are also used. , and A therapy that combines three types of internal radiation therapy. teeth, The goal is to reduce dosage and side effects while increasing the effectiveness of tumor treatment. Expected .

[0006] on the other hand So, As a thromboembolic agent for medical therapy and as a thromboembolic agent for radiotherapy Disadvantages of Lipiodol Therefore, it is necessary to implement a combination of three treatments: embolization, chemotherapy, and internal radiation therapy. eye, Stabilization of labeling for the radionuclide iodine, and for chemotherapeutic agents Inclusion Stabilization , and The design and preparation of novel microsphere formulations that form permanent embolic effects against tumor blood vessels are necessary. Simultaneously, emission computed tomography or single-photon emission computed tomography using gamma rays emitted from radionuclides is required to obtain real-time in vivo distribution data of the microspheres. in Dynamic monitoring allows for the development of integrated diagnostic and therapeutic formulations.

[0007] Building upon the foundations and current state of existing technologies, the present invention provides a microsphere embolization agent that integrates chemotherapy, internal radiation therapy, and embolization, and can be used in emission computed tomography or single-photon emission computed tomography, integrating diagnostic and therapeutic processes. formulation It can carry various chemotherapeutic agents and can be labeled with the radionuclide iodine, and the labeling is highly stable. Compared with chemoembolic microspheres or radiotherapy embolic microsphere preparations, the embolic preparation disclosed in this invention, which combines chemotherapy and radiotherapy, significantly reduces the doses of chemotherapeutic agents and therapeutic radionuclides that need to be delivered when achieving equivalent therapeutic effects. Let Ta. [Overview of the project]

[0008] The objective of this invention is to address the foundation and current state of existing technologies regarding tumors. chemistry Radiation therapy Transembolization therapy, and Imaging for The objective is to provide a multifunctional microsphere formulation and a method for preparing the same. This microsphere uses a polyvinyl alcohol derivative as a backbone material and is polymerized and crosslinked with an N-acryloyl amino acid monomer to cure. created These microspheres can not only be labeled with the radioactive nuclide iodine, but can also adsorb chemotherapeutic agents. This multifunctional microsphere formulation enables tumor embolization and combined therapy with radiotherapy and chemotherapy, while simultaneously allowing emission computed tomography or single-photon emission computed tomography.

[0009] Polyvinyl alcohol derivative used in the microsphere formulation according to the present invention polymer The monomers are as follows: general formula It is ,

[0010] [ka]

[0011] Of these, the molecular weight range is 10 kDa ~ The polyvinyl alcohol derivative is 1000 kDa, preferably 75 kDa. polymer The monomer was formed by an acetal reaction between the hydroxyl group of N-(2,2-dimethoxyethyl)-2-acrylamide and polyvinyl alcohol. The degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide is in the range of 0.1 ~ 40%, preferably 10% ~ It is 15%.

[0012] The N-acryloyl amino acid monomer added to the microsphere formulation according to the present invention has a structure according to the following formula,

[0013] [ka]

[0014] Of these, R is one or more of the following structures:

[0015] [ka]

[0016] Preferably, it is N-acryloyltyrosine.

[0017] The molar ratio range of N-acryloyl amino acid monomer and polyvinyl alcohol derivative polymer monomer contained in the microsphere formulation according to the present invention is 50:1 ~The ratio is 1000:1, preferably 245:1. ~ The ratio is 260:1.

[0018] The present invention provides a method for preparing the microsphere formulation described above, the preparation procedure being as follows. (1) Aqueous solution of N-acryloyl amino acid monomer, aqueous solution of polyvinyl alcohol derivative polymer monomer and past Prepare potassium sulfate aqueous solutions. death ; (2) Prepare an oil phase solution containing an emulsifier or dispersant. And, at this time The oil phase consists of liquid paraffin, butyl acetate, soybean oil, or silicone oil, and the emulsifier is Span-80, Span-60, or cellulose acetate / butyrate. the law of nature ; (3) Add the aqueous solution to the oil phase drop by drop to form an emulsion. death ; (4) Catalyst N,N,N',N'-tetramethylethylene Ji An aqueous solution containing an amine is added to the emulsion, catalyzing a polymerization crosslinking reaction and causing it to harden, forming microspheres. death ; (5) After washing the microspheres, sieve them to collect microspheres of different particle sizes. 。

[0019] vinegar The procedure for preparing the solution containing the N-acryloyl amino acid monomer of step (1) is as follows: As follows: Dissolve N-acryloyl amino acids using a sodium hydroxide solution. Yes. At this time, The concentration of the sodium hydroxide solution is 200 ~ A concentration of 400 mg / mL is preferred, with a volume of 0.1 ~ 20 mL is preferred. stomach. Furthermore In contrast, concentrated hydrochloric acid by Adjust the pH to neutral; the mass of N-acryloyl amino acids is 0.1 ~ 20 g is preferred.

[0020] The concentration of the polyvinyl alcohol derivative polymer monomer solution in step (1) is 7.5 ~20% is preferred, and its volume is 2 ~ 200 mL is preferred.

[0021] The concentration of the potassium persulfate solution in step (1) is 100. ~ A concentration of 200 mg / mL is preferred, with a volume of 0.05 ~ 10 mL is preferred.

[0022] The oil phase in step (2) is liquid paraffin, soybean oil, or silicone oil, preferably liquid paraffin. The volume of liquid paraffin used is 10 ~ 1000 mL is preferred. The emulsifier is Span-80 or Span-60, preferably Span-80. The dose of Span-80 is 0.05 ~ 15 g is preferred.

[0023] Step (4) N,N,N',N'-tetramethylethylene Ji The volume of the amine solution is 0.2 ~ 20 mL is preferred, and its concentration is 5%. ~ 15% (v / v) is preferred.

[0024] Step (4) By catalyst Polymerization and crosslinking Response The temperature is 25 ~ 60°C is preferred, and the reaction time is 2 ~ 12 hours is preferable.

[0025] The washing procedure in step (5) was as follows: washing was performed twice with each solvent in the order of ethyl acetate, anhydrous ethanol, and ultrapure water.

[0026] Step (5) At After sieving the microspheres of The particle size range of microspheres is 、 20 ~ 120 μm, 120 ~ 200 μm, 200 ~ 300 μm, 300 ~ 500 μm, 500 ~ 700 μm, 700 ~ 900 μm, 900~ 1100 μm, 1100 ~ It is 1300 μm, preferably 20 ~ It is 120 μm.

[0027] The microsphere formulation according to the present invention is 、 at the same time Below It can be labeled with one or more of the following radioactive iodine nuclides: iodine-123, iodine-125, and iodine-131, preferably iodine-131.

[0028] release Iodine, a radioactive nuclide by sign The following procedure Do it this way: (1) Suspend the microspheres in phosphate buffer; (2) Add the sodium iodide solution of the radioactive nuclide iodine to the microsphere suspension from step (1) and mix thoroughly; (3) Add the phosphate buffer containing chloramine-T to the mixture from step (2) and shake gently to mix thoroughly; (4) 25 ~ 10 ~ Allow to react for 60 minutes; (5) The microsphere suspension from step (4) is centrifuged and washed, and the precipitate is radioactive iodine-labeled microspheres.

[0029] vinegar The pH of the phosphate buffer in TEP(1) is 7.4, and its volume is 0.1 ~ 10 mL is preferred, and the mass of the microsphere powder is 5 ~ 500 mg is preferred.

[0030] The activity of the radioactive iodine solution in step (2) is 10 ~ 100 mCi / mL is preferred, in addition ru The volume of the radioactive iodine solution is 0.01 ~ 10 mL is preferred.

[0031] Step (3) At In addition ru The volume of chloramine-T solution is 0.01 ~ 10 mL is preferred, and its concentration is 1 ~ A 10 mg / mL concentration is preferred.

[0032] The water bath temperature in step (4) is 25 ~ 45°C is preferred, and the labeling time is 10 ~ 60 minutes is preferable.

[0033] In this invention evening Microsphere formulations are release The labeling efficiency after labeling with radioactive iodine was 81%. ~ It's 99.5%.

[0034] In this invention evening Microspheres formulation This is achieved by ion exchange or adsorption, using one or the following chemotherapeutic agents. multiple Can carry different types of drugs: Adriamycin hydrochloride, Epirubicin, Daunorubicin, Mitoxantrone, Irinotecan, Topotecan and Preferably, it is Adriamycin hydrochloride.

[0035] In this invention evening Microsphere preparations labeled with radioactive iodine. did The procedure for radioactive microspheres to carry chemotherapeutic agents is as follows: below That is correct.

[0036] (1) Dissolve the chemotherapeutic agent in ultrapure water to prepare a solution of the chemotherapeutic agent.

[0037] (2) Step (1) twist The obtained solution is added to the microsphere suspension or radionuclide iodine-labeled microsphere suspension prepared by the method described above, and shaken several times. ~ Wait 30 minutes.

[0038] (3) After centrifuging the mixed solution from step (2), the supernatant is removed to obtain drug-supported microspheres.

[0039] Book Microsphere formulation according to the invention and radioactive microspheres labeled with the radioactive nuclide iodine The chemotherapeutic agent is administered by the procedure described above. Carrying, The amount of drug carried Adriamycin hydrochloride but 10% ~ 60%, Epirubicin but 7% ~ 45%, daunorubicin but 10% ~ 43%, Mitoxantrone but 9% ~ 35%, Irinotecan but 8% ~ 30%, Topotecan but 5% ~ twenty five% That is .

[0040] In this invention evening Microspheres formulation Its monomer composition also includes 2-acrylamido-2-methylpropylsulfonic acid monomer. During polymerization crosslinking reaction to, N-acryloyl amino acid monomers and polyvinyl alcohol derivative polymer monomers are further combined with 2-acrylamido-2-methylpropylsulfonic acid monomers. Three molar ratio ranges of N-acryloyl amino acids, 2-acrylamido-2-methylpropylsulfonic acid, and polyvinyl alcohol derivative polymer monomers. teeth Preferably 50:50:1 ~ The ratio is 1000:1000:1.

[0041] mentioned above The preparation procedure for the microsphere formulation is as follows: (1) Prepare an N-acryloyl amino acid solution; (2) Prepare a solution of 2-acrylamido-2-methylpropylsulfonic acid; (3) Prepare a solution of polyvinyl alcohol derivative polymer monomer; (4) Prepare a potassium persulfate solution; (5) Mix the solutions from steps (1), (2), (3), and (4) uniformly to form the aqueous phase; (6) The aqueous phase of step (5) 、 Add the oil phase containing the dispersant and stir under nitrogen; (7) Prepare an N,N,N',N'-tetramethylethylenediamine solution; (8) Add the solution from step (7) to the mixed solution from step (6) under nitrogen, and continue 1 ~ Allow to react for 10 hours. (9) After centrifuging the reaction mixture from step (8), wash the precipitate and disperse it in ultrapure water; (10) The precipitated solution from step (9) is sieved by a wet method to obtain microspheres of different particle sizes.

[0042] In step (1), preferably 200 ~ Dissolve N-acryloyl amino acids in a 400 mg / mL sodium hydroxide solution. Yes. At this time, The volume of the sodium hydroxide solution is 0.1 ~ 20 mL is preferred. stomach. Adjust the pH to neutral with concentrated hydrochloric acid; the mass of N-acryloyl amino acid is 0.1 ~ 20 g is preferred.

[0043] In step (2), the mass of 2-acrylamido-2-methylpropylsulfonic acid is 0.1 ~ 20 g is preferred, dissolved in ultrapure water, and the volume used is 0.1 g. ~ 20 mL is preferred.

[0044] In step (3), the concentration of the polyvinyl alcohol derivative polymer monomer solution is 10% ~ 21% is preferred, and the volume used is 1 ~ 200 mL is preferred.

[0045] The concentration of the potassium persulfate solution in step (4) is 100. ~ A concentration of 200 mg / mL is preferred, and the volume used is 0.05 mg / mL. ~ 10 mL is preferred.

[0046] The oil phase in step (6) is preferably butyl acetate, of Used body The product is 30 ~ 3000 mL is preferred, cellulose acetate butyrate is preferred as the dispersant, and the amount of cellulose acetate butyrate used is 0.3 ~ 30 g is preferred.

[0047] In step (7), N,N,N',N'-tetramethylethylene Ji Amine solution of Used body The product is preferably 0.3 to 25 mL, and its concentration is 6%. ~ A 17% (v / v) ratio is preferred.

[0048] The water bath temperature in step (8) is 45 ~ 65°C is preferred.

[0049] Regarding the washing procedure in step (9), preferably, the washing is performed sequentially twice with ethyl acetate, anhydrous ethanol, and ultrapure water in that order.

[0050] After sieving in step (10) of The particle size range of the microspheres is 20 ~ 120 μm, 120 ~ 200 μm, 200 ~ 300 μm, 300 ~ 500 μm, 500 ~ 700 μm, 700 ~ 900 μm, 900 ~ 1100 μm, 1100 ~ It is 1300 μm, preferably 20 ~ It is 120 μm.

[0051] In this invention evening Microspheres formulation It also contains N,N'-methylenediacrylamide as a monomer component. ru. the edgeThe molar ratio range of the three polymer monomers, N-acryloyl amino acids, N,N'-methylenediacrylamide, and polyvinyl alcohol derivatives, is 50:5:1. ~ A ratio of 1000:200:1 is preferable. Furthermore, as mentioned above The procedure for preparing the microsphere formulation is as follows:

[0052] (1) Prepare an N-acryloyl amino acid solution; (2) Prepare an N,N'-methylenediacrylamide solution; (3) Prepare a solution of polyvinyl alcohol derivative polymer monomer; (4) Prepare a potassium persulfate solution.

[0053] (5) Mix the solutions from steps (1), (2), (3), and (4) uniformly to form the aqueous phase; (6) Add the aqueous phase from step (5) to the oil phase containing the emulsifier and stir under nitrogen to emulsify; (7) N,N,N',N'-tetramethylethylene Ji Prepare an amine solution; (8) Add the solution from step (7) to the emulsion from step (6) under nitrogen and continue to react in a water bath for 6 to 24 hours; (9) After centrifuging the emulsion from step (8), wash the precipitate and disperse it in ultrapure water; (10) The precipitated solution from step (9) is sieved by a wet method to obtain microspheres of different particle sizes.

[0054] In step (1), preferably 200 ~ Dissolve N-acryloyl amino acids in a 400 mg / mL sodium hydroxide solution. Do so. At that time, The volume of the sodium hydroxide solution is 0.1 ~ 20 mL is preferred. stomach. Adjust the pH to neutral with concentrated hydrochloric acid; the mass of N-acryloyl amino acid is 0.1 ~ 20 g is preferred.

[0055] Step (2) InThe mass of N,N'-methylenediacrylamide is 0.01 ~ 5 g is preferred stomach. With ultrapure water Dissolution After that, its volume is 0.1 ~ 15 mL is preferred.

[0056] The concentration of the polyvinyl alcohol derivative polymer monomer solution in step (3) is 7.5%. ~ 20% is preferred, and The polyvinyl alcohol derivative polymer monomer solution used The volume is 1 ~ 200 mL is preferred.

[0057] The concentration of the potassium persulfate solution in step (4) is 100. ~ 200 mg / mL is preferred, and The potassium persulfate solution to be used The volume is 0.05 ~ 10 mL is preferred.

[0058] The oil phase in step (6) is liquid paraffin, soybean oil, or silicone oil, preferably liquid paraffin; to use The volume of liquid paraffin is 10 ~ 1000 mL is preferred; the emulsifier should be Span-80 or Span-60. and Preferably, the span is -80, and the dose is 0.05 ~ 15 g is preferred.

[0059] The N,N,N',N'-tetramethylethylene used in step (7) Ji The volume of the amine solution is 0.2 ~ 20 mL is preferred, and its concentration is 5 ~ 15% (v / v) is preferred.

[0060] The water bath temperature in step (8) is 30 ~ 50°C is preferred.

[0061] The cleaning procedure in step (9) involves washing twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water in that order.

[0062] The particle size range after sieving the microspheres in step (10) is 20 ~ 120 μm, 120 ~ 200 μm, 200 ~ 300 μm, 300 ~ 500 μm, 500 ~ 700 μm, 700 ~ 900 μm, 900 ~ 1100 μm, 1100 ~ 1300 μm, preferably 20 ~ It is 120 μm.

[0063] The microsphere formulation labeled with the radionuclide iodine-125 or iodine-131 according to the present invention is used for liver cancer, liver metastases, uterine fibroids, kidney cancer, or pancreatic cancer. against Embolization therapy, radiation therapy and It can be used in combination therapy with chemotherapy. Furthermore, Radioactive nuclide iodine-131 Therefore, Emission computed tomography or single-photon emission computed tomography implement It is possible.

[0064] The microsphere formulation labeled with the radionuclide iodine-123 according to the present invention is used for embolization therapy of liver cancer, liver metastases, uterine fibroids, kidney cancer, or pancreatic cancer. and It can be used in combination therapy with chemotherapy. Furthermore, The radioactive nuclide iodine-123 can be analyzed using emission computed tomography or single-photon emission computed tomography. [Brief explanation of the drawing]

[0065] [Figure 1] This is a field emission scanning electron microscope image of a DOX-NAT-PVA microsphere (A) and its cross-section (B). [Figure 2] This is a particle size distribution diagram of DOX-NAT-PVA microspheres. [Figure 3]These are radiochromatographic scanning patterns of 131I-DOX-NAT-PVA microspheres: (A) when the sample was placed on paper immediately after labeling the 131I-DOX-NAT-PVA microspheres, and (B) when the sample was placed on paper after centrifuging and washing the 131I-DOX-NAT-PVA microspheres. The labeling efficiency is 92.39% ± 1.51%. [Figure 4] This figure shows the label stability of 131I-NAT-PVA microspheres and 131I-DOX-NAT-PVA microspheres in phosphate buffer (PBS) and 50% fetal bovine serum (FBS). [Figure 5] Figures (A, B) show the measurement and comparison of the IC50 values ​​of DOX in DOX-NAT-PVA microspheres and 131I-DOX-NAT-PVA microspheres after 72 hours of incubation; Figures (C, D) show the measurement and comparison of the IC50 values ​​of 131I in 131I-NAT-PVA microspheres and 131I-DOX-NAT-PVA microspheres (mean ± SD, n=3, **p<0.01). [Figure 6] Figures (A, B) show a comparison of the measured IC50 values ​​of DOX in DOX-NAT-PVA microspheres and 131I-DOX-NAT-PVA microspheres under oxygen deficiency; Figures (C, D) show a comparison of the measured IC50 values ​​of 131I in 131I-NAT-PVA microspheres and 131I-DOX-NAT-PVA microspheres (mean ± SD, n=3, ***p<0.001). [Figure 7] These are SPECT / CT images of a model rat at a predetermined time after hepatic artery embolization (arrows indicate the location of microspheres). h: hours; d: days. [Figure 8](A) Model rats of liver cancer orthotopic transplantation in each treatment group were injected with 18F-FDG into the tail vein before hepatic artery embolization (Pre) and on days 1, 3, 7, and 14 after hepatic artery embolization, and the changes in 18F-FDG uptake at the tumor site were monitored. These are small animal PET / CT images (arrows indicate tumor location, and the dosage for each treatment group is as follows: Control group: 0.2 mL physiological saline, NAT-PVA group: 0.2 mL NAT-PVA microsphere suspension (containing 0.04 mL microspheres), 131Ilow-NAT-PVA group: 0.2 mL 131I-NAT-PVA microsphere suspension (containing 0.04 mL microspheres and 200 μCi 131I), 131Ihigh-NAT-PVA group: 0.2 mL 131I-NAT-PVA microsphere suspension (containing 0.04 mL microspheres and 500 μCi 131I) (B) The DOXlow-NAT-PVA group was injected with 0.2 mL DOX-NAT-PVA microsphere suspension (0.04 mL microspheres, containing 0.5 mg DOX), the DOXhigh-NAT-PVA group was injected with 0.2 mL DOX-NAT-PVA microsphere suspension (0.04 mL microspheres, containing 2.5 mg DOX), and the 131Ilow-DOXlow-NAT-PVA group was injected with 0.2 mL 131I-DOX-NAT-PVA microsphere suspension (0.04 mL microspheres, containing 0.5 mg DOX and 200 μCi 131I); (B) The curves showing the time course of the maximum 18F-FDG uptake (SUVmax) at the tumor site in each treatment group (mean ± SD, n = 5. *p<0.05, **p<0.01, ***p<0.001); (C) In vitro images of tumors 14 days after transhepatic artery embolization in each treatment group. d. Day. [Modes for carrying out the invention]

[0066] The present invention will be further described below with reference to examples and drawings. [Examples]

[0067] 120 mg ofTake 120 μL of N-acryloyltyrosine (NAT). of Dissolve in a 400 mg / mL sodium hydroxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 40 mg of Take N,N'-methylenediacrylamide and disperse it in 150 μL of ultrapure water using ultrasound, then add another 1.334 mL of 7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ 13%) was added and dissolved by ultrasound. The N-acryloyltyrosine solution was added to the polyvinyl alcohol derivative solution and mixed uniformly, then 87 μL was added. of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.12 g of 8 mL containing Span-80 of In addition to the liquid paraffin, the mixture was emulsified by stirring at 600 rpm for 10 minutes under nitrogen in a 37°C water bath. Then, 200 μL was dispensed. of 10% (v / v) N,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT-PVA microsphere powder.

[0068] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 131Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 30 minutes. We did After labeling is complete, take 5 μL of the microsphere solution and perform radiopaper chromatography. law to twist The labeling efficiency of microspheres quantitative The remaining labeled microspheres of Centrifugation is performed at 5000 rpm for 5 minutes, the supernatant is removed, and ultrapure water is then added to the lower precipitate. inside No radioactivity detected I want to Wash thoroughly, then add another 5 μL of The microsphere solution was taken and the radiochemical purity of the microspheres after washing was measured using radiopaper chromatography.

[0069] 15 mg of 131 Prepare I-NAT-PVA microspheres and 200 μL of The solution was suspended in ultrapure water. Approximately 7.5 mg of adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0070] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Having (Figure 1). The average particle size of the microspheres was approximately 51.4 ± 3.3 μm (Figure 2). As a result by Then, the drug loading and encapsulation efficiency of the microspheres were 33.25% ±1.73% and 99.76% ± 0.18%, respectively. Radio paper chromatography experimental results showed that the labeling rate of the microspheres was 92.59% ± 1.51%, and the radiochemical purity was 99.46% ± 0.47% (Figure 3). In vitro labeling stability In evaluation , on day 31 at still maintained a radiochemical purity of more than 85% (Figure 4).

[0071] Under normoxic conditions 131 After co-incubating I-DOX-NAT-PVA microspheres with N1S1 rat liver cancer cells for 72 hours, the IC of DOX and 131 I 50 was measured As a result, which were 3.74 ± 0.240 ng / mL and 8.15 ± 0.521 μCi / mL, respectively. The IC of DOX in the control group DOX-NAT-PVA microspheres 50 was 9.26 ± 0.127 ng / mL, and the IC of I in the control group 131 I-NAT-PVA microspheres 131 I 50 was 19.56 ± 1.190 μCi / mL That was the case. The Rera results from , 131 I-DOX-NAT-PVA microspheres suggested that DOX chemotherapy and 131 I radiotherapy had a synergistic effect, with a synergy index of 0.82 (Figure 5). Also, Under hypoxic conditions 131 After incubating I-DOX-NAT-PVA microspheres with N1S1 cells for 72 hours, the IC of DOX and 131 I 50 was measured As a result, which were 、 4.20 ± 1.14 ng / mL and 6.68 ± 1.25 μCi / mL, respectively. The IC of DOX in the control group DOX-NAT-PVA microspheres 50 was 37.98 ± 2.58 ng / mL, and the control group 131 I-NAT-PVA microspheres131 I IC 50 The value was 57.12 ± 7.41 μCi / mL. . So Rera As a result from , 131 DOX chemotherapy with I-DOX-NAT-PVA microspheres 131 The synergistic effect of radiotherapy was suggested, with a synergistic index of 0.23 (Figure 6).

[0072] Results from in vivo animal experiments if via the hepatic artery 131 I-NAT-PVA microspheres transplanted orthotopically into N1S1 liver cancer of Injecting into rats By doing Microspheres under SPECT / CT imaging of small animals of Model rats Okeru This suggests that the distribution was accurately monitored and accurate liver tumor images could be provided up to 31 days after injection (Figure 7). 18 Intravenous injection of F-FDG was performed, and N1S1 orthotopic transplanted liver cancer was identified using PET / CT imaging of small animals. of The growth of tumors in rats was monitored after treatment using various methods. Fruit, The median survival time for rats in the untreated group was 20 days (n = 5). Show It was done. 131 I-DOX-NAT-PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 Embolization therapy was performed using a combination therapy with a dose of 200 μCi for I and a dose of 1.67 mg / kg for DOX. Then, 14 days later of Tumor tissue 18 The F-FDG image is negative. of They showed that they would continue observation until day 60. Then, Rat death teeth Not observed (n = 5). Control group 131 I-NAT-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 I. Embolization treatment was performed using high-dose radiotherapy with a radiation dose of 500 μCi. Then, 14 days later of Tumor tissue 18 The F-FDG image is negative. of They showed that they would continue observation until day 60. Then, Rat death teeth Not observed (n = 5). Control group 131 I-NAT-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization was performed using low-dose radiotherapy with a radiation dose of 200 μCi. However, 14 days later of Tumor tissue 18 F-FDG image from tumor but recurrence What I did Show It was done. The median survival time was 23 days (n = 5). The control group received NAT-PVA microspheres (microsphere dose of 0.14 mL / kg) alone. in Embolization treatment was performed using this method. However, 14 days later of Tumor tissue 18 F-FDG image from tumor but recurrence What I did Show So The median survival time was 22 days (n = 5). these As a result twist , 131 I-DOX-NAT-PVA Microspheres Using this method, The combined effect of radiotherapy and chemotherapy in vivo on tumor treatment Show , 200μCi 131 Dosages of 1 and 1.67 mg / kg DOX provide equivalent therapeutic effect to 500 μCi of radiotherapy embolization or 8.35 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let This was confirmed (Figure 8).

[0073] Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 500 μCi (I). Then, rear ofThe median survival time in rats was 21 days (n = 5); they were treated with embolization using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.35 mg / kg). However, rear of The median survival time for rats was 22 days (n = 5). DC bead TM Embolization therapy was performed using chemotherapy-embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.35 mg / kg). However, rear of The median survival time for rats was 45 days (n = 5). [Examples]

[0074] 150 mg of Take 150 μL of N-acryloylhistidine (NAH). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 35 mg of N,N'-methylenediacrylamide was taken and dispersed in 150 μL of ultrapure water using ultrasound, and then 1.334 mL of a 7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) was added. ~ A 13% solution was added and dissolved by sonication. The N-acryloylhistidine solution was added to the polyvinyl alcohol derivative solution and mixed uniformly, then 100 μL was added. of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.12 g of Add 8 mL of liquid paraffin containing Span-80, and emulsify by stirring at 600 rpm for 10 minutes in a 40°C water bath under a nitrogen atmosphere. Then, 200 μL of 10% (v / v )N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution ofThe liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAH-PVA microsphere powder.

[0075] 10 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (20 μL) of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 15 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. Then, place the microsphere suspension in a 40°C water bath and label for 35 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 90.13% ± 3.11%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 99.75% ± 1.63%.

[0076] 15 mg of 131 Prepare I-NAH-PVA microspheres and produce 200 μL. of The solution was suspended in ultrapure water. Approximately 5.5 mg of Adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. AfterThe mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0077] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Having The average particle size of the microspheres was approximately 54.9 ± 5.7 μm. As a result by Then The drug load and encapsulation rate in the microspheres were 25.39% ± 2.37% and 94.23% ± 0.35%, respectively. In the in vitro labeling stability study, radiochemical purity of over 80% was maintained even on day 31.

[0078] Results from in vivo animal experiments if via the hepatic artery 131 I-DOX-NAH-PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 (The dose of I was 220 μCi, and the dose of DOX was 1.82 mg / kg) for N1S1 orthotopic liver cancer of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Control group 131 I-NAH-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using high-dose radiotherapy with a dose of 650 μCi (of I). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NAH-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 8.65 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group131 I-NAH-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 220 μCi (I). When I did that, rear of The median survival time in rats was 23 days (n = 5). Embolization was performed using DOX-NAH-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 1.82 mg / kg, low-dose chemotherapy) in the control group. When I did that, rear of The median survival time for rats was 22 days (n = 5). The control group received NAH-PVA microspheres (microsphere dose of 0.14 mL / kg) alone. in Embolization treatment using When I did that, rear of The median survival time for rats was 22 days (n = 5). these As a result twist , 131 I-DOX-NAH-PVA microspheres are in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 220 μCi 131 Doses of 1 and 1.82 mg / kg DOX provided equivalent therapeutic effect to 650 μCi of radiotherapy embolization or 8.65 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 650 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.65 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TMEmbolization therapy using chemotherapy-embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.65 mg / kg) When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0079] 150 mg of Take 170 μL of N-acryloyltryptophan (NATP). of Dissolve in 400 mg / mL sodium hydroxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 45 mg of Take N,N'-methylenediacrylamide and sonicate 200 μL of Disperse in ultrapure water, then add 1.334 mL of 7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ A 13% solution was added and dissolved by sonication. The N-acryloyl tryptophan solution was added to the polyvinyl alcohol derivative solution and mixed uniformly, then 95 μL was added. of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.12 g of Span -80 Contains To 8 mL of liquid paraffin Add The mixture was emulsified in a 45°C water bath under a nitrogen atmosphere by stirring at 600 rpm for 10 minutes. Then, 200 μL was used. of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered.~ Collect the precipitate between 70 μm sieves, perform dehydration and centrifugation twice with acetone, and dry it overnight at room temperature in a vacuum dryer to obtain dried NATP-PVA microsphere powder.

[0080] 15 mg of Take microspheres, add 100 μL of 0.01 M PBS with pH 7.4 to suspend the microspheres. Take 20 μL of 10 mCi / mL Na 131 I solution and add it to the microsphere suspension, mix evenly, then add 20 μL of 10 mg / mL chloramine-T in PBS, shake gently to mix .the , then place the microsphere suspension in a 40°C water bath for 40 minutes of labeling We did . After labeling is completed, the labeling rate of the microspheres is measured to be 89.24% ± 1.27%. Add 1 mL of ultrapure water to the remaining labeled microsphere solution, centrifuge at 5000 rpm for 5 minutes, remove the supernatant, continue adding ultrapure water to the lower precipitate, until no radioactivity is detected in the supernatant I want to , then stop washing. The radiochemical purity of the lower-layer microspheres is measured to be 99.64%±1.42%.

[0081] 25 mg of 131 I-NATP-PVA microspheres are prepared, and suspended in 200 μL of ultrapure water. Take about 12 mg of adriamycin hydrochloride, dissolve it in 200 μL of ultrapure water. Add the adriamycin solution to the microsphere suspension, shake gently to suspend and mix thoroughly. Then stand still, and shake once every 5 minutes I made them do it. After about 15 minutes, the color of the supernatant no longer changes After , centrifuge at 5000 rpm for 5 minutes, and collect the supernatant. The precipitate of is further washed three times repeatedly with ultrapure water.

[0082] Under a field emission scanning electron microscope, There, the microspheres are ,largesize It is uniform, with a round surface Yes . The average particle diameter of the microspheres was approximately 45.29±5.9 μm. According to measurement As a result results if , the drug loading capacity and encapsulation efficiency of the microspheres were 30.13% ± 1.73% and 92.76% ± 0.18%, respectively. In the study on in vitro labeling stability, on day 31 at still maintained a radiochemical purity of over 80%.

[0083] According to the results of in vivo animal experiments if , via the hepatic artery 131 I-DOX-NATP-PVA microspheres (microsphere dose: 0.14 mL / kg, 131 I dose: 252 μCi, DOX dose: 1.89 mg / kg) were injected into rats with N1S1 orthotopic transplanted liver cancer of rats for embolization therapy When I did that, within 60 days after Below the treatment teeth no rat death was observed (n = 5). In the control group 131 I-NATP-PVA radiotherapy embolization microspheres (microsphere dose: 0.14 mL / kg, 131 I dose: 642 μCi, high-dose radiotherapy) were used for embolization therapy When I did that, within 60 days after Below the treatment teeth no rat death but was observed (n = 5). In the control group treated with DOX-NATP-PVA chemotherapy embolization microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 9.25 mg / kg, high-dose chemotherapy) for embolization therapy When I did that, within 60 days after Below the treatment teeth no rat death but was observed (n = 5). In the control group 131 I-NATP-PVA radiotherapy embolization microspheres (microsphere dose: 0.14 mL / kg, 131 I dose: 252 μCi, low-dose radiotherapy) were used for embolization therapy When I did that, after ofThe median survival time in rats was 23 days (n = 5). Embolization was performed using DOX-NATP-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 1.89 mg / kg, low-dose chemotherapy) in the control group. When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization was performed using NATP-PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NATP-PVA microspheres Using this method, in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 252μCi 131 Doses of 1 and 1.89 mg / kg DOX were found to have an effect equivalent to 642 μCi of radiotherapy embolization or 9.25 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 642 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 9.25 mg / kg) When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0084] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of 400 mg / m L waterDissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 150 mg of Take 2-acrylamido-2-methylpropylsulfonic acid and sonicate 200 μL of Dissolved in ultrapure water. 5 mL each of N-acryloyltyrosine solution and 2-acrylamido-2-methylpropylsulfonic acid solution. of 15% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 67 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 60 μL of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.3 g of Adding 30 mL of butyl acetate containing cellulose acetate butyrate, the mixture was emulsified in a 55°C water bath under a nitrogen atmosphere by stirring at 1000 rpm for 10 minutes. Then, 200 μL of 10% (v / v) N,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution of The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was removed. The precipitate was then washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Afterward, the precipitate was dispersed with a small amount of ultrapure water, and each portion was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT-AMPS-PVA microsphere powder.

[0085] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. 5 μL of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 5 μL of10 mg / mL chloramine-T PBS solution was added, and the mixture was gently shaken to mix evenly. the After that, the microsphere suspension was placed in a 37°C water bath for labeling for 30 minutes We did . After completion of labeling, the labeling rate of the microspheres was measured to be 85.41% ± 2.54%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, followed by centrifugation at 5000 rpm for 5 minutes, the supernatant was removed, ultrapure water was continuously added to the lower precipitate, and no radioactivity was detected in the supernatant I want to until washing was completed. The radiochemical purity of the lower-layer microspheres was measured to be 99.27% ± 0.62%.

[0086] 15 mg of 131 I-NAT-AMPS-PVA microspheres were prepared and suspended in 200 μL of ultrapure water. About 15 mg of adriamycin hydrochloride was taken and dissolved in 200 μL of ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and mixed thoroughly. Then, the mixture was allowed to stand still, and shaken once every 5 minutes I made them do it. After about 15 minutes, the color of the supernatant stopped changing After , followed by centrifugation at 5000 rpm for 5 minutes, and the supernatant was collected. The precipitate of was further washed three times repeatedly with ultrapure water.

[0087] Under a field emission scanning electron microscope There, the microspheres had ,large size It is uniform, round surfaces Yes . The average particle size of the microspheres was about 49.3 ± 6.5 μm. According to measurement As a result results Then , the drug loading and encapsulation efficiency of the microspheres were 40.13% ± 2.72% and 80.2% ± 0.38%, respectively. For in vitro labeling stability In evaluation , on the 31st day at a radiochemical purity of more than 82% was still maintained.

[0088] According to the results of in vivo animal experiments, via the hepatic artery131 I-DOX-NAT-AMPS-PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 I The dose was 282 μCi, and the dose of DOX was 2.41 mg / kg) for N1S1 orthotopic liver cancer. of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Control group 131 I-NAT-AMPS-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using high-dose radiotherapy (with a dose of 629 μCi). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NAT-AMPS-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 9.42 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group 131 I-NAT-AMPS-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 282 μCi (I). When I did that, rear of The median survival time in rats was 23 days (n = 5). Embolization was performed using DOX-NAT-AMPS-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 2.41 mg / kg, low-dose chemotherapy) in the control group. When I did that, rear of The median survival time for rats was 23 days (n = 5). Embolization was performed using NAT-AMPS-PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NAT-AMPS-PVA microspheres are in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 282μCi 131 Doses of 1 and 2.41 mg / kg DOX were found to have an effect equivalent to 629 μCi of radiotherapy embolization or 9.42 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 629 μCi (I). When I did that, The median survival time of the rats afterward was 22 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 9.42 mg / kg). When I did that, The median survival time of the rats afterward was 23 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 9.42 mg / kg) When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0089] 190 mg of Take 190 μL of N-acryloylhistidine (NAH). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 150 mg of Take 2-acrylamido-2-methylpropylsulfonic acid and extract 500 μL using ultrasound. of Dissolved in ultrapure water. 5 mL each of N-acryloylhistidine solution and 2-acrylamido-2-methylpropylsulfonic acid solution. of15% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 67 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 80 μL of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.3 g of 30 mL containing cellulose acetate butyrate of In addition to butyl acetate, the mixture was emulsified in a 55°C water bath under a nitrogen atmosphere by stirring at 1000 rpm for 10 minutes. Then, 200 μL was dispensed. of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 5 hours. After the reaction was complete, the reaction solution of The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was removed. The precipitate was then washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Afterward, the precipitate was dispersed with a small amount of ultrapure water, and each portion was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAH-AMPS-PVA microsphere powder.

[0090] 10 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. 15 μL of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 15 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 40 minutes. We didAfter labeling was complete, the labeling efficiency of the microspheres was measured to be 92.62% ± 1.71%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 99.41% ± 1.22%.

[0091] 15 mg of 131 Prepare I-NAH-AMPS-PVA microspheres and produce 200 μL. of The solution was suspended in ultrapure water. Approximately 15 mg of Adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0092] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 56.2 ± 3.8 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 50.1% ± 0.26% and 99.58% ± 0.15%, respectively. In the discussion of in vitro label stability, on day 31... at It also maintained a radiochemical purity of over 88%.

[0093] Results from in vivo animal experiments show that via the hepatic artery 131 I-DOX-NAH-AMPS-PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 Iwas 282 μCi, and the dosage of DOX was 2.41 mg / kg) was injected into N1S1 orthotopic transplanted liver cancer of rats, and within 60 days after embolization therapy When I did that, 60 days after Below within teeth no rat death was observed (n = 5). In the control group 131 I-NAH-AMPS-PVA radiotherapy embolization microspheres (microsphere dosage: 0.14 mL / kg, 131 I dose: 648 μCi, high-dose radiotherapy) were used for embolization therapy When I did that, 60 days after Below within teeth rat death but was not observed (n = 5). In the control group, DOX-NAT-AMPS-PVA chemotherapy embolization microspheres (microsphere dosage: 0.14 mL / kg, DOX dosage: 10.27 mg / kg, high-dose chemotherapy) were used for embolization therapy When I did that, 60 days after Below within teeth rat death but was not observed (n = 5). In the control group 131 I-NAH-AMPS-PVA radiotherapy embolization microspheres (microsphere dosage: 0.14 mL / kg, 131 I dose: 282 μCi, low-dose radiotherapy) were used for embolization therapy When I did that, after of the median survival time of rats was 23 days (n = 5). In the control group, DOX-NAH-AMPS-PVA chemotherapy embolization microspheres (microsphere dosage: 0.14 mL / kg, DOX dosage: 2.41 mg / kg, low-dose chemotherapy) were used for embolization therapy However, after of the median survival time of rats was 23 days (n = 5). In the control group, NAH-AMPS-PVA microspheres (microsphere dosage: 0.14 mL / kg) were used alone for embolization therapy When I did that, the median survival time of the rats was 22 days (n = 5). these the results twist , 131 I-DOX-NAH-AMPS-PVA microspheres Using in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 282μCi 131 Doses of 1 and 2.41 mg / kg DOX were found to have an effect equivalent to 648 μCi of radiotherapy embolization or 10.27 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 648 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 10.27 mg / kg). When I did that, The median survival time of the rats afterward was 23 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 10.27 mg / kg). When I did that, rear of The median survival time for rats was 48 days (n = 5). [Examples]

[0094] 200 mg of Take 200 μL of N-acryloyltryptophan (NATP). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 230 mg of 2-acrylamido-2-methylpropylsulfonic acid was taken and dissolved in 500 μL of ultrapure water using ultrasound. 5 mL of the N-acryloyltryptophan solution and 2-acrylamido-2-methylpropylsulfonic acid solution were then added. of 15% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 67 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 60 μL of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.3 g of Adding 30 mL of butyl acetate containing cellulose acetate butyrate, the mixture was emulsified in a 55°C water bath under a nitrogen atmosphere by stirring at 1000 rpm for 10 minutes. Then, 200 μL of 10% (v / v) N,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 5 hours. After the reaction was complete, the reaction solution of The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was removed. The precipitate was then washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Afterward, the precipitate was dispersed with a small amount of ultrapure water, and each portion was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of a 70 μm mesh was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NATP-AMPS-PVA microsphere powder.

[0095] 15 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (25 μL) of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 20 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 40 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 85.62% ± 3.72%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 99.62% ± 1.92%.

[0096] 25 mg of 131 Prepare powder of I-NATP-AMPS-PVA microspheres, add 200 μL of and suspend the powder in ultrapure water. Take about 30 mg of doxorubicin hydrochloride, dissolve it in 200 μL of of ultrapure water. Add the doxorubicin solution to the microsphere suspension, shake gently to suspend and mix thoroughly. Then allow the mixture to stand, and shake once every 5 minutes I made them do it. After about 15 minutes, the color of the supernatant no longer changes After ; centrifuge at 5000 rpm for 5 minutes and collect the supernatant. The precipitate of is repeatedly washed three more times with ultrapure water.

[0097] Under a field emission scanning electron microscope, the There, microspheres ,large have size It is uniform, and round surfaces Yes . The average particle size of the microspheres was about 53.58 ± 3.94 μm. According to measurement As a result results Then , the drug loading capacity and encapsulation efficiency of the microspheres were 51.4% ± 4.23% and 83.76% ± 3.62%, respectively. In the study of in vitro labeling stability, radiochemical purity of over 85% was still maintained on day 31 at .

[0098] According to the results of in vivo animal experiments, via the hepatic artery 131 I-DOX-NATP-AMPS-PVA microspheres (microsphere dose: 0.14 mL / kg, 131 I radioactivity dose: 282 μCi, DOX dose: 2.41 mg / kg) were injected into N1S1 orthotopic transplanted liver cancer of rats for embolization therapy When I did that, within 60 days after the treatment Below , teeth no rat death was observed (n = 5). In the control group 131 I-NATP-AMPS-PVA radiotherapy embolization microspheres (microsphere dose: 0.14 mL / kg, 131Embolization treatment using high-dose radiotherapy (with a dose of 629 μCi). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NATP-AMPS-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 9.42 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group 131 I-NATP-AMPS-PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 282 μCi (low-dose radiotherapy). When I did that, rear of The median survival time in rats was 23 days (n = 5). The control group was treated with embolization using DOX-NATP-AMPS-PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 2.41 mg / kg, low-dose chemotherapy). However, rear of The median survival time for rats was 22 days (n = 5). Embolization was performed using NATP-AMPS-PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NATP-AMPS-PVA microspheres Using this method, in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 282μCi 131 Doses of 1 and 2.41 mg / kg DOX were found to have an effect equivalent to 629 μCi of radiotherapy embolization or 9.42 mg / kg of chemotherapy embolization. Show , significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 629 μCi (I). When I did that, The median survival time of the rats afterward was 22 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 9.42 mg / kg). When I did that, The median survival time of the rats afterward was 22 days (n = 5). DCbead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 9.42 mg / kg). When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0099] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 2 mL of 7.5% Polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 100 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.15 g of Add 10 mL of liquid paraffin containing Span-80, and emulsify by stirring at 600 rpm for 10 minutes in a 55°C water bath under a nitrogen atmosphere. Then, 200 μL of 20% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution ofThe liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT / PVA microsphere powder.

[0100] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 30 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 94.37% ± 1.71%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 99.68% ± 1.29%.

[0101] 15 mg of 131 Prepare I-NAT / PVA microspheres and 200 μL of The solution was suspended in ultrapure water. Approximately 7.5 mg of adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. AfterThe mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0102] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 55.3 ± 6.3 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 32.51% ± 2.52% and 97.51% ± 1.63%, respectively. In the discussion of in vitro label stability, on day 31... at It also maintained a radiochemical purity of over 81%.

[0103] Results from in vivo animal experiments show that via the hepatic artery 131 I-DOX-NAT / PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 I The dose was 240 μCi, and the dose of DOX was 1.74 mg / kg) for N1S1 orthotopic liver cancer. of Inject into rats for embolization treatment. When I did that, 60 days later Below No rat deaths were observed within the control group (n = 5). 131 I-NAT / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using high-dose radiotherapy with a dose of 540 μCi (of I). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NAT / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 8.73 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group131 I-NAT / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 240 μCi (low-dose radiotherapy). When I did that, rear of The median survival time in rats was 23 days (n = 5). The control group was treated with embolization using DOX-NAT / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 1.74 mg / kg, low-dose chemotherapy). However, rear of The median survival time in rats was 22 days (n = 5). Embolization was performed using NAT / PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NAT / PVA Microspheres Using this method, in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 240μCi 131 Doses of 1 and 1.74 mg / kg DOX produced an effect equivalent to 540 μCi of radiotherapy embolization or 8.73 mg / kg of chemotherapy embolization. Show ...significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 540 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.73 mg / kg). When I did that, The median survival time of the rats afterward was 22 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.73 mg / kg). When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0104] 200 mg of Take 200 μL of N-acryloylhistidine (NAH). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 2.5 mL of 7.5% Polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 150 μL. of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.15 g of Add 10 mL of liquid paraffin containing Span-80, and emulsify by stirring at 600 rpm for 10 minutes in a 40°C water bath under a nitrogen atmosphere. Then, 250 μL of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAH / PVA microsphere powder.

[0105] 10 mg of Take a microsphere and add 100 μL ofThe solution was suspended in 0.01 M pH 7.4 PBS. (20 μL) of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 15 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 35 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 92.13% ± 2.03%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 95.89% ± 2.43%.

[0106] 15 mg of 131 I-NAH / PVA microspheres were prepared and suspended in 200 μL of ultrapure water. Approximately 6.5 mg of adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0107] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 57.7 ± 2.7 μm. As a result by ThenThe drug load and encapsulation rate in the microspheres were 25.39% ± 2.37% and 84.23% ± 3.41%, respectively. In the in vitro labeling stability study, radiochemical purity of over 80% was maintained even on day 31.

[0108] Results from in vivo animal experiments show that via the hepatic artery 131 I-DOX-NAH / PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 I The dose was 245 μCi, and the dose of DOX was 1.96 mg / kg) for N1S1 orthotopic liver cancer. of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Control group 131 I-NAH / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using high-dose radiotherapy with a dose of 635 μCi (I). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NAH / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 9.25 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group 131 I-NAH / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 245 μCi (I). When I did that, rear of The median survival time in rats was 23 days (n = 5). Embolization was performed using DOX-NAH / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 1.96 mg / kg, low-dose chemotherapy) in the control group. When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization was performed using NAH / PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NAH / PVA Microspheres Using this method, in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show , 245μCi 131 Doses of 1 and 1.96 mg / kg DOX produced an effect equivalent to 635 μCi of radiotherapy embolization or 9.25 mg / kg of chemotherapy embolization. Show ...significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 635 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 9.25 mg / kg). When I did that, The median survival time of the rats afterward was 22 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 9.25 mg / kg). When I did that, The median survival time of the rats afterward was 46 days (n = 5). [Examples]

[0109] 250 mg of Take N-acryloyltryptophan (NATP) and add 250 μL. of 400 mg / m L water It was dissolved in sodium oxide solution, and the pH was adjusted to neutral with concentrated hydrochloric acid. 3 mL of7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 145 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.20 g of 15 mL containing Span-80 of In addition to the liquid paraffin, the mixture was emulsified by stirring at 600 rpm for 10 minutes in a 45°C water bath under a nitrogen atmosphere. Then, 300 μL was dispensed. of 10% (v / v) N,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of a 70 μm mesh was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NATP / PVA microsphere powder.

[0110] 20 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. 15 μL of 10 mCi / mL Na 131 Take solution I and add it to the microsphere suspension, mix uniformly, then add 20 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 30 minutes. We didAfter labeling was complete, the labeling efficiency of the microspheres was measured to be 86.24% ± 1.82%. 1 mL of ultrapure water was added to the remaining labeled microsphere solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 96.58% ± 1.63%.

[0111] 15 mg of 131 Prepare I-NATP / PVA microspheres and 200 μL of The solution was suspended in ultrapure water. Approximately 5 mg of Adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0112] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 49.29 ± 2.9 μm. As a result by Then The drug load and encapsulation rate in the microspheres were 20.43% ± 1.73% and 81.72% ± 2.31%, respectively. In the in vitro labeling stability study, radiochemical purity of over 85% was maintained even on day 31.

[0113] Results from in vivo animal experiments if via the hepatic artery 131 I-DOX-NATP / PVA microspheres (microsphere dosage is 0.14 mL / kg) 131 IThe dose was 270 μCi, and the dose of DOX was 1.95 mg / kg) for N1S1 orthotopic liver cancer. of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Control group 131 I-NATP / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using high-dose radiotherapy with a dose of 650 μCi (of I). When I did that, 60 days later Below inside teeth Rat death but No cases observed (n = 5). Embolization was performed using DOX-NATP / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 8.90 mg / kg, high-dose chemotherapy) in the control group. When I did that, 60 days later Below inside teeth Rat death but Not observed (n = 5). Control group 131 I-NATP / PVA radiotherapy embolic microspheres (microsphere dose is 0.14 mL / kg) 131 Embolization treatment using low-dose radiotherapy with a dose of 270 μCi (low-dose radiotherapy). When I did that, rear of The median survival time in rats was 23 days (n = 5). Embolization was performed using DOX-NATP / PVA chemotherapy-embolic microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 1.95 mg / kg, low-dose chemotherapy) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). Embolization was performed using NATP / PVA microspheres alone (microsphere dosage: 0.14 mL / kg) in the control group. When I did that, The median survival time of the rats afterward was 22 days (n = 5). these As a result twist , 131 I-DOX-NATP / PVA microspheres Using in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy GoodThe effect Show , 270μCi 131 Doses of 1 and 1.95 mg / kg DOX produced an effect equivalent to 650 μCi of radiotherapy embolization or 8.90 mg / kg of chemotherapy embolization. Show ...significantly reduced the dosage of radiotherapy and chemotherapy. Let We confirmed that this was the case. Also, 131 I-Lipiodol (The dosage of Lipiodol is 0.67 mL / kg) 131 Embolization treatment using a dose of 650 μCi (I). When I did that, The median survival time of the rats afterward was 21 days (n = 5). Embolization was performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.90 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.90 mg / kg). When I did that, rear of The median survival time for rats was 46 days (n = 5). these As a result twist , 131 I-DOX-NATP / PVA microspheres Using this method, in vivo Release Tumor treatment using a combination of radiation therapy and chemotherapy Good The effect Show ...significantly reduced the dosage of radiotherapy and chemotherapy. Let I confirmed that this was the case. [Examples]

[0114] 120 mg of Take 120 μL of N-acryloyltyrosine (NAT). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 40 mg of Take N,N'-methylenediacrylamide and sonicate 150 μL of Disperse in ultrapure water, then add 1.334 mL of7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ 13%) was added and dissolved by ultrasound. The N-acryloyltyrosine solution was added to the polyvinyl alcohol derivative solution and mixed uniformly, and 87 μL was added to it. of Add 150 mg / mL potassium persulfate solution, mix well, then add 0.12 g of Add 8 mL of liquid paraffin containing Span-80 and emulsify by stirring at 600 rpm for 10 minutes in a 37°C water bath under a nitrogen atmosphere. Then, 200 μL of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT / PVA microsphere powder.

[0115] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 123 Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 35 minutes. We didAfter labeling was complete, the labeling rate of the microspheres was measured to be 94.59% ± 1.31%. 1 mL of ultrapure water was added to the remaining microsphere labeling solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 96.62% ± 2.92%.

[0116] 15 mg of 123 Prepare I-NAT-PVA microspheres and 200 μL of The solution was suspended in ultrapure water. Approximately 7.5 mg of adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0117] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 51.2 ± 2.5 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 31.35% ± 2.42% and 98.16% ± 0.41%, respectively. In the in vitro labeling stability study, the radiochemical purity remained above 83% even on day 31.

[0118] Results from in vivo animal experiments if In small animal SPECT / CT images, model rats Okeru 123 The distribution of I-DOX-NAT-PVA microspheres is accurately monitored. Ta. via the hepatic artery 123I-DOX-NAT-PVA microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 8.61 mg / kg) in N1S1 orthotopic liver cancer of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT-PVA microspheres alone (microsphere dose: 0.14 mL / kg) in the control group. When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.61 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.61 mg / kg). When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0119] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of Dissolve in 400 mg / mL sodium hydroxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 150 mg of Take 2-acrylamido-2-methylpropylsulfonic acid and sonicate 200 μL of Dissolved in ultrapure water. 5 mL each of N-acryloyltyrosine solution and 2-acrylamido-2-methylpropylsulfonic acid solution. of 15% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 67 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ After adding to the 13% solution and mixing uniformly, add to 60 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the Next, 30 mL of butyl acetate containing 0.3 g of cellulose acetate butyrate was added, and the mixture was emulsified by stirring at 1000 rpm for 10 minutes in a 55°C water bath under a nitrogen atmosphere. Then, 200 μL of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution of The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was removed. The precipitate was then washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Afterward, the precipitate was dispersed with a small amount of ultrapure water, and each portion was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT-AMPS-PVA microsphere powder.

[0120] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. 5 μL of 10 mCi / mL Na 123 Take solution I and add it to the microsphere suspension, mix uniformly, then add 5 μL of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 30 minutes. We did After labeling was complete, the labeling rate of the microspheres was measured to be 85.82% ± 1.63%. 1 mL of ultrapure water was added to the remaining microsphere labeling solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 96.53% ± 2.14%.

[0121] 15 mg of123 Prepare I-NAT-AMPS-PVA microspheres and produce 200 μL. of The solution was suspended in ultrapure water. Approximately 15 mg of Adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. After The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0122] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 52.7 ± 3.5 μm. Then The drug load and encapsulation rate of the microspheres were 45.43% ± 2.61% and 90.86% ± 1.29%, respectively. In the in vitro labeling stability study, the radiochemical purity was maintained at over 84% even on day 31.

[0123] Results from in vivo animal experiments showed that in model rats under SPECT / CT imaging of small animals... Okeru 123 The distribution of I-DOX-NAT-AMPS-PVA microspheres is accurately monitored. Ta. via the hepatic artery 123 I-DOX-NAT-AMPS-PVA microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 9.21 mg / kg) for N1S1 orthotopic liver cancer transplantation of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT-AMPS-PVA microspheres alone (microsphere dose: 0.14 mL / kg) in the control group. When I did that, rear ofThe median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 9.21 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 9.21 mg / kg). When I did that, rear of The median survival time for rats was 46 days (n = 5). [Examples]

[0124] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 2 mL of 7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ Add to the 13% solution, mix uniformly, then add to 100 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.15 g of 10 mL containing Span-80 of In addition to the liquid paraffin, the mixture was emulsified by stirring at 600 rpm for 10 minutes in a 55°C water bath under a nitrogen atmosphere. Then, 200 μL was dispensed. of 20% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution ofThe liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT / PVA microsphere powder.

[0125] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 123 Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 40°C water bath and labeled for 35 minutes. We did After labeling was complete, the labeling rate of the microspheres was measured to be 95.39% ± 0.62%. 1 mL of ultrapure water was added to the remaining microsphere labeling solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 99.52% ± 2.53%.

[0126] 15 mg of 123 Prepare I-NAT / PVA microsphere powder and produce 200 μL. of The solution was suspended in ultrapure water. Approximately 7.5 mg of adriamycin hydrochloride was taken and added to 200 μL. of It was dissolved in ultrapure water. The adriamycin solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, and vibrated once every 5 minutes. I made them do it. After about 15 minutes, the color of the supernatant stops changing. AfterThe mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0127] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 54.4 ± 3.4 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 33.26% ± 3.31% and 96.26% ± 2.37%, respectively. In vitro label stability was also observed. In evaluation On the 31st day at It also maintained a radiochemical purity of over 83%.

[0128] Results from in vivo animal experiments showed that in model rats under SPECT / CT imaging of small animals... Okeru 123 The distribution of I-NAT / PVA microspheres is accurately monitored. Ta. via the hepatic artery 123 I-DOX-NAT / PVA microspheres (microsphere dose: 0.14 mL / kg, DOX dose: 8.97 mg / kg) in N1S1 orthotopic liver cancer of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT / PVA microspheres alone in the control group (microsphere dose: 0.14 mL / kg). When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-DOX emulsion (Lipiodol dose: 0.67 mL / kg, DOX dose: 8.97 mg / kg). When I did that, The median survival time of the rats afterward was 22 days (n = 5). DC bead TMEmbolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, DOX dose: 8.97 mg / kg). When I did that, rear of The median survival time for rats was 45 days (n = 5). [Examples]

[0129] 120 mg of Take 120 μL of N-acryloyltyrosine (NAT). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 40 mg of Take N,N'-methylenediacrylamide and sonicate 150 μL of Disperse in ultrapure water, then add 1.334 mL of A 7.5% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%~13%) was added and subsequently dissolved by sonication. N-acryloyltyrosine solution was added to the polyvinyl alcohol derivative solution and mixed uniformly, then 87 μL was added. of Add 150 mg / mL potassium persulfate solution and mix thoroughly. .the After, 0.12 g of Add 8 mL of liquid paraffin containing Span-80 and emulsify by stirring at 600 rpm for 10 minutes in a 37°C water bath under a nitrogen atmosphere. Then, 200 μL of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 12 hours. After the reaction was complete, the reaction solution ofThe mixture was centrifuged at 5000 rpm for 5 minutes to remove the liquid paraffin, and the lower precipitate was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. The precipitate was then dispersed with a small amount of ultrapure water and sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. The precipitate between the 40 and 70 μm sieves was collected, dehydrated twice with acetone and centrifuged, and then dried overnight in a vacuum dryer at room temperature to obtain dried NAT-PVA microsphere powder.

[0130] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 125 Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 30 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 90.41% ± 2.61%. 1 mL of ultrapure water was added to the remaining microsphere labeling solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 98.96% ± 2.51%.

[0131] 15 mg of 125 Prepare I-NAT-PVA microsphere powder and produce 200 μL. of It was suspended in ultrapure water. Approximately 7.5 mg of irinotecan was taken and added to 200 μL. of The irinotecan solution was dissolved in ultrapure water. The irinotecan solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, vibrated once every 5 minutes, and after approximately 30 minutes, centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. Precipitation. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0132] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 52.7 ± 4.6 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 20.15% ± 2.11% and 61.27% ± 2.71%, respectively. In the in vitro labeling stability study, radiochemical purity of over 85% was maintained even on day 31.

[0133] In in vivo animal experiments, model rats were subjected to SPECT / CT imaging of small animals. Okeru 125 The distribution of I-IRI-NAT-PVA microspheres is accurately monitored. Ta. via the hepatic artery 125 I-IRI-NAT-PVA microspheres (microsphere dose: 0.14 mL / kg, IRI dose: 25.61 mg / kg) for N1S1 orthotopic liver cancer transplantation of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT-PVA microspheres alone in the control group (microsphere dose: 0.14 mL / kg). When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-IRI emulsion (Lipiodol dose: 0.67 mL / kg, IRI dose: 25.61 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, IRI dose: 25.61 mg / kg) When I did that, 60 days later Below inside teeth Rat death but It could not be seen (n = 5). [Examples]

[0134] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of 400 mg / m L water Dissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 150 mg of Take 2-acrylamido-2-methylpropylsulfonic acid and sonicate 200 μL of Dissolved in ultrapure water. 5 mL each of N-acryloyltyrosine solution and 2-acrylamido-2-methylpropylsulfonic acid solution. of 15% polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 67 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ After adding to the 13% solution and mixing uniformly, add to 60 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.3 g of 30 mL containing cellulose acetate butyrate of In addition to butyl acetate, the mixture was emulsified in a 55°C water bath under a nitrogen atmosphere by stirring at 1000 rpm for 10 minutes. Then, 200 μL was dispensed. of 10% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution of The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was removed. The precipitate was then washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Afterward, the precipitate was dispersed with a small amount of ultrapure water, and each portion was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT-AMPS-PVA microsphere powder.

[0135] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. 5 μL of 10 mCi / mL Na 125 Take solution I and add it to the microsphere suspension, mix uniformly, then add 5 μL of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 30 minutes. We did After labeling was complete, the labeling efficiency of the microspheres was measured to be 89.54% ± 2.71%. 1 mL of ultrapure water was added to the remaining microsphere labeling solution, and the mixture was centrifuged at 5000 rpm for 5 minutes. The supernatant was removed, and ultrapure water was subsequently added to the precipitate. Radioactivity was not detected in the supernatant. I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 96.59% ± 3.62%.

[0136] 15 mg of 125 Prepare I-NAT-AMPS-PVA microspheres and produce 200 μL. of It was suspended in ultrapure water. Approximately 15 mg of irinotecan was taken and added to 200 μL. of The irinotecan solution was dissolved in ultrapure water. The irinotecan solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, vibrated once every 5 minutes, and after approximately 30 minutes, centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. Precipitation. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0137] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 55.27 ± 0.31 μm. As a result by ThenThe drug load and encapsulation rate in the microspheres were 32.69% ± 3.51% and 65.4% ± 2.74%, respectively. In the in vitro labeling stability study, radiochemical purity of over 80% was maintained even on day 31.

[0138] In in vivo animal experiments, model rats were subjected to SPECT / CT imaging of small animals. Okeru 125 The distribution of I-IRI-NAT-AMPS-PVA microspheres is accurately monitored. Ta. via the hepatic artery 125 I-IRI-NAT-AMPS-PVA microspheres (microsphere dose: 0.14 mL / kg, IRI dose: 29.77 mg / kg) for N1S1 orthotopic liver cancer transplantation of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT-AMPS-PVA microspheres alone (microsphere dose: 0.14 mL / kg) in the control group. When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-IRI emulsion (Lipiodol dose: 0.67 mL / kg, IRI dose: 29.77 mg / kg). When I did that, The median survival time of the rats afterward was 21 days (n = 5). DC bead TM Embolization therapy using chemotherapy embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, IRI dose: 29.77 mg / kg) When I did that, 60 days later Below inside teeth Rat death but It could not be seen (n = 5). [Examples]

[0139] 170 mg of Take 170 μL of N-acryloyltyrosine (NAT). of 400 mg / m L waterDissolve in sodium oxide solution and adjust the pH to neutral with concentrated hydrochloric acid. 2 mL of 7.5% Polyvinyl alcohol derivative (molecular weight of polyvinyl alcohol: 75 kDa, degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide: 12%) ~ After adding to the 13% solution and mixing uniformly, add to 100 μL of Add 150 mg / mL potassium persulfate solution and mix well. Match Ta .the After, 0.15 g of 10 mL containing Span-80 of In addition to the liquid paraffin, the mixture was emulsified by stirring at 600 rpm for 10 minutes in a 55°C water bath under a nitrogen atmosphere. Then, 200 μL was dispensed. of 20% (v / v ) N ,N,N',N'-tetramethylethylene Ji The amine aqueous solution was added drop by drop. The reaction mixture was stirred under nitrogen for a further 4 hours. After the reaction was complete, the reaction solution of The liquid paraffin was removed by centrifugation at 5000 rpm for 5 minutes, and the precipitate at the bottom was washed twice sequentially with ethyl acetate, anhydrous ethanol, and ultrapure water. Then, the precipitate was dispersed with a small amount of ultrapure water, and each was sieved through 70 μm and 40 μm sieves in that order. The residue through the 70 μm sieve and the residue through the 40 μm sieve were filtered. ~ The precipitate between the sieves of 70 μm was collected, and after dehydration by centrifugation twice with acetone, it was dried overnight in a vacuum dryer at room temperature to obtain a dried NAT / PVA microsphere powder.

[0140] 5 mg of Take a microsphere and add 100 μL of The solution was suspended in 0.01 M pH 7.4 PBS. (10 μL) of 10 mCi / mL Na 125 Take solution I and add it to the microsphere suspension, mix uniformly, then add 10 μL. of Add 10 mg / mL chloramine-T PBS solution and shake gently to mix. .the Next, the microsphere suspension was placed in a 37°C water bath and labeled for 45 minutes. We didAfter labeling was complete, the labeling rate of the microspheres was measured to be 92.02% ± 3.98%. 1 mL of the remaining microsphere labeling solution was added. of Ultrapure water is added, the mixture is centrifuged at 5000 rpm for 5 minutes, the supernatant is removed, and ultrapure water is added to the lower precipitate. If radioactivity is not detected in the supernatant, I want to The material was washed to this extent. The radiochemical purity of the lower layer of microspheres was measured to be 98.22% ± 3.81%.

[0141] 15 mg of 125 Prepare I-NAT / PVA microsphere powder and produce 200 μL. of It was suspended in ultrapure water. Approximately 7.5 mg of irinotecan was taken and added to 200 μL. of The irinotecan solution was dissolved in ultrapure water. The irinotecan solution was added to the microsphere suspension, gently shaken to suspend, and thoroughly mixed. Then, it was allowed to stand, vibrated once every 5 minutes, and after approximately 35 minutes, centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. Precipitation. of Furthermore, it was washed three times repeatedly with ultrapure water.

[0142] Under a field emission scanning electron microscope There, Microspheres ,large Kisa It is uniform, Round surface Yes The average particle size of the microspheres was approximately 53.0 ± 2.5 μm. As a result by Then The drug load and encapsulation rate of the microspheres were 19.03% ± 2.31% and 58.81% ± 4.81%, respectively. In vitro label stability was also observed. In evaluation On the 31st day at It also maintained a radiochemical purity of over 80%.

[0143] In in vivo animal experiments, model rats were subjected to SPECT / CT imaging of small animals. Okeru 125 The distribution of I-IRI-NAT / PVA microspheres is accurately monitored. Ta. via the hepatic artery 125I-IRI-NAT / PVA microspheres (microsphere dose: 0.14 mL / kg, IRI dose: 35.8 mg / kg) for N1S1 orthotopic liver cancer transplantation of Inject into rats for embolization treatment. When I did that, 60 days later Below inside teeth No rat deaths were observed (n = 5). Embolization was performed using NAT / PVA microspheres alone in the control group (microsphere dose: 0.14 mL / kg). When I did that, rear of The median survival time for rats was 22 days (n = 5). Embolization therapy was also performed using Lipiodol-IRI emulsion (Lipiodol dose: 0.67 mL / kg, IRI dose: 35.8 mg / kg). When I did that, The median survival time of the rats afterward was 22 days (n = 5). DC bead TM Embolization therapy using chemotherapy-embolic microspheres (microsphere dose: 0.18 mL / kg, particle size: 75-150 μm, IRI dose: 35.8 mg / kg) When I did that, 60 days later Below inside teeth Rat death but It could not be seen (n = 5).

Claims

1. It is a microsphere formulation, Its characteristics are that it is formed by the polymerization and crosslinking of an N-acryloyl amino acid monomer represented by formula (1) and a polyvinyl alcohol derivative polymer monomer represented by formula (2) in a constant proportion. 【Chemistry 1】 Of these, R is one or more of the following structures: 【Chemistry 2】 Among these, the polyvinyl alcohol derivative polymer monomer represented by formula (2) has a molecular weight range of 10 kDa to 1000 kDa, and is formed by the reaction of N-(2,2-dimethoxyethyl)-2-acrylamide with the hydroxyl group of polyvinyl alcohol, with a degree of substitution of N-(2,2-dimethoxyethyl)-2-acrylamide ranging from 0.1% to 40%; The molar ratio range of the N-acryloyl amino acid monomer and the polyvinyl alcohol derivative polymer monomer is 50:1 to 1000:1; A microsphere formulation that can be labeled with the radioactive nuclide iodine and simultaneously carry a chemotherapeutic agent.

2. A method for producing a microsphere formulation according to claim 1, comprising the following steps: (a) Prepare aqueous solutions of N-acryloyl amino acid monomer, polyvinyl alcohol derivative polymer monomer, and potassium persulfate; (b) Prepare an oil phase solution containing an emulsifier or dispersant, wherein the oil phase is liquid paraffin, butyl acetate, soybean oil, or silicone oil, the emulsifier is Span-80 or Span-60, and the dispersant is cellulose acetate butyrate; (c) Add the aqueous solution to the oil phase drop by drop to form an emulsion; (d) An aqueous solution containing the catalyst N,N,N',N'-tetramethylethylenediamine is added to the emulsion, catalyzing the polymerization crosslinking reaction and causing it to harden and form microspheres; (e) A manufacturing method comprising washing and then sieving microspheres to recover microspheres of different particle size ranges.

3. A microsphere formulation according to claim 1, wherein the radioactive nuclide iodine is one or more of the following radioactive isotopes of iodine: iodine-123, iodine-125, iodine-131.

4. A method for producing a microsphere preparation according to claim 1, wherein the labeling rate when labeled with the radioactive nuclide iodine is 81% to 99.5%, and the labeling procedure is as follows: (a) Suspend the microspheres in phosphate buffer; (b) Add the sodium iodide solution containing the radionuclide iodine described in claim 3 to the microsphere suspension of (a) and mix uniformly; (c) Add the phosphate buffer containing chloramine-T to the mixture from (b) and shake gently to combine; (d) Allow the reaction mixture to react in a water bath at 25–45°C for 10–60 minutes; A method for producing (e) microspheres, wherein the microsphere suspension of (d) is centrifuged and washed, and the precipitate is radioactive nuclide iodine-labeled microspheres.

5. A microsphere formulation according to claim 1, wherein the chemotherapeutic agent is one or more of the following drugs: adriamycin hydrochloride, epirubicin, daunorubicin, mitoxantrone, irinotecan, topotecan.

6. A method for producing a microsphere formulation according to claim 1, wherein the microsphere or the prepared radionuclide iodine-labeled microspheres are loaded with the chemotherapeutic agent according to claim 5 by ion exchange or adsorption, the amount of the drug loaded is 10% to 60%, and the loading procedure is as follows: (a) Dissolve the chemotherapeutic agent in ultrapure water to prepare a solution of the chemotherapeutic agent; (b) Add the solution from (a) to the prepared microsphere suspension or the prepared radionuclide iodine-labeled microsphere suspension, shake several times, and wait for 5 to 30 minutes; (c) A manufacturing method comprising centrifuging the mixed solution of (b), removing the supernatant, and obtaining drug-supported microspheres.

7. A microsphere formulation according to claim 1, wherein the particle size range is 20 to 1300 μm.

8. A method for producing a microsphere formulation according to claim 1, wherein, during the polymerization crosslinking, 2-acryloyl amino acid monomer is further added to the N-acryloyl amino acid monomer and the polyvinyl alcohol derivative polymer monomer, and the molar ratio of the three—N-acryloyl amino acid monomer, 2-acryloyl amino acid monomer, and polyvinyl alcohol derivative polymer monomer—is in the range of 50:50:1 to 1000:1000:

1.

9. A method for producing a microsphere formulation according to claim 1, wherein, during the polymerization crosslinking, an N,N'-methylenediacrylamide monomer is further added to the N-acryloyl amino acid monomer and the polyvinyl alcohol derivative polymer monomer, and the molar ratio of the three—N-acryloyl amino acid monomer, N,N'-methylenediacrylamide monomer, and polyvinyl alcohol derivative polymer monomer—is in the range of 50:5:1 to 1000:200:

1.

10. A microsphere formulation according to claim 3, wherein the microsphere formulation is labeled with 125I and / or 131I, 125 I and / or 131 Microsphere formulations labeled with I are used in tumor embolization, chemotherapy, radiotherapy, and emission computed tomography or single-photon emission computed tomography.

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