Patent Application
20250135051
The present application claims priority to Chinese Patent Application No. 202311388190.4, entitled “YTTRIUM [90Y] MICROSPHERE AND PREPARATION METHOD THEREFOR AND USE THEREOF”, and filed to the China National Intellectual Property Administration on Oct. 25, 2023, the entire contents of which are incorporated herein by reference.
The present application relates to the technical field of radiopharmaceutical preparation, specifically relates to an yttrium [90Y] microsphere and preparation method therefor and use thereof.
Interventional radiotherapy technology is an effective method for treating tumor developed in recent years. Yttrium [90Y] as a radiotherapy has the following advantages: high energy, short half-life, short penetration distance in the body, and easy protection. Yttrium [90Y] microspheres injection precision interventional radiotherapy is a technology that integrates interventional and precision internal radiation therapy. Yttrium [90Y] radioactive microspheres, as an important treatment for liver cancer, will hopefully provide a powerful weapon for the comprehensive treatment of liver cancer patients in our country, which can improve their cure rate and prolong the overall survival.
Yttrium-90 glass microspheres and styrene divinylbenzene copolymer microspheres have been used for the treatment of liver cancer. However, these microspheres have the defect of insufficient adsorption strength for radioactive elements, resulting in poor therapeutic effect on tumors.
Therefore, the technical problem to be solved in the present application is to overcome the defect of insufficient adsorption strength of yttrium [90Y] microspheres in the prior art, so as to provide an yttrium [90Y] microspheres and preparation method therefor and use thereof.
For this purpose, the present application provides an yttrium [90Y] microsphere, wherein the yttrium [90Y] microsphere comprises a polystyrene sulfonic acid type resin microsphere and yttrium-90 adsorbed within the polystyrene sulfonic acid type resin microsphere.
Further, the polystyrene sulfonic acid type resin microsphere is prepared by reacting a polystyrene microsphere with a sulfonating agent.
Further, the sulfonating agent is selected from one or more of liquid sulfur trioxide, concentrated sulfuric acid, chlorosulfonic acid, or sulfite; preferably, the sulfonating agent is concentrated sulfuric acid.
Wherein, the mass percentage of concentrated sulfuric acid in the present application is greater than or equal to 70%, such as 98%.
Further, the polystyrene microsphere is prepared by using styrene as a monomer and divinylbenzene as a crosslinking agent, mixing and reacting with an aqueous solution containing a suspending agent in the presence of an initiator and pore forming agent.
Further, the pore forming agent is selected from one or more of toluene and n-heptane; and/or, the suspension agent is polyvinyl alcohol; and/or, the initiator is benzoyl peroxide.
Further, the preparation method of the polystyrene sulfonic acid type resin microsphere comprises:
Further, during the preparation of polystyrene microspheres, the temperature of heating reaction is 80° C. to 100° C., and the time is 2 hours to 5 hours. Stirring is performed at 40° C. to 60° C. overnight before the heating reaction. The time of shearing emulsification is 3 min to 10 min.
Further, during the preparation of polystyrene sulfonic acid type resin microspheres, adding polystyrene microspheres to concentrated sulfuric acid at 0° C. to 4° C., and heating to 37° C. to 45° C. for reaction.
Further, the heating reaction also comprises the steps of suction filtration, washing, drying and sieving.
Further, the polystyrene microsphere is made of the following raw materials:
Further, the yttrium [90Y] microsphere has a radioactive activity ranging from 2.5 GBq/g to 25 GBq/g.
Further, the mass percentage of polyvinyl alcohol in the aqueous solution containing polyvinyl alcohol is in a range from 0.5% to 3% (e.g., 1%, 2%, or 0.8%).
Further, the yttrium [90Y] sulfonic acid type resin microsphere has a particle size ranging from 10 μM to 60 μM, preferably 28 μm to 35 μm, more preferably 30 μm to 35 μm.
The present application also provides a method for preparing the yttrium [90Y] microsphere above, wherein the method comprises:
Further, the ratio of radioactive activity of yttrium chloride [90Y] solution, the volume of yttrium sulfate-89 solution, and the mass of polystyrene sulfonic acid type resin microspheres is 30-200 GBq: 30-200 ml: 5-15.5 g. Preferably, the concentration of yttrium sulfate-89 solution is in a range from 1 g/L to 9.5 g/L.
Wherein, the yttrium chloride [90Y] solution has a concentration ranging from 1 mg/ml to 5 mg/ml.
Further, the ratio of the volume of buffer salt solution and the mass of polystyrene sulfonic acid type resin microspheres is 70-300 ml: 5-15.5 g.
Further, the buffer salt has a concentration ranging from 2 g/l to 60 g/l.
Further, the buffer salt solution is selected from one or more of sodium phosphate solution, sodium dihydrogen phosphate solution, disodium hydrogen phosphate solution, potassium phosphate solution, potassium dihydrogen phosphate solution, dipotassium hydrogen phosphate solution, PBS solution, sodium acetate and Tris-HCl buffer solution. Optionally, in the S2 step, sodium phosphate solution is used first for washing, and then sodium dihydrogen phosphate solution is used for washing.
Further, in the S2 step, water, sodium phosphate solution, sodium dihydrogen phosphate solution, and water are used sequentially for washing.
Further, the sodium dihydrogen phosphate solution has a concentration ranging from 2 g/L to 25 g/L; the sodium phosphate solution has a concentration ranging from 10 g/L to 60 g/L.
The present application also provides a pharmaceutical preparation of yttrium [90Y] microsphere, comprising any one of the yttrium [90Y] microsphere above or yttrium [90Y] microsphere prepared by any one of the methods above, and also a pharmaceutically acceptable excipient. Optionally, the pharmaceutical acceptable excipient is a solvent.
Wherein, the solvent can be distilled water, water for injection, or glucose aqueous solution and other conventional solvents. For the pharmaceutical preparation of yttrium [90Y] microspheres, in the unit dose, the radioactive activity of yttrium [90Y] microspheres is ranging from 3 GBq to 20 GBq.
Further, the pharmaceutical preparation is an injection, and the radioactive activity of the injection per unit dose is ranging from 3 GBq to 20 GBq.
The unit dose of the injection comprises 0.8 g to 4 g of the yttrium [90Y] microspheres.
The preparation method of yttrium [90Y] microspheres injection in the present application can also comprise routine intermediate radioactivity detection, encapsulation, sterilization or product activity detection.
Wherein, the filling volume of each injection bottle is 5 ml when encapsulation.
The present application also provides the use of any of the above-mentioned yttrium [90Y] microspheres or yttrium [90Y] microspheres prepared by any one of the above-mentioned preparation methods or any of the above-mentioned pharmaceutical preparations in the preparation of medicaments for preventing or treating liver cancer.
The raw material (yttrium chloride [90Y] solution) is commercially available, and its radioactive activity can also be customized. According to the activity meter operation instructions, measure and record the activity of incoming raw materials (yttrium chloride [90Y] solution) to ensure that the activity of radioactive raw materials is consistent with the target activity during production. The formula for calculating the activity of the required radioactive raw material is: A=KA0/e−0.26t, wherein k is the process loss coefficient, A0 is the activity value at the calibration time, A is the radioactive activity value at the determination time, and t is the difference in days between the determination time and the calibration time.
The technical solution of the present application has the following advantages:
1. The yttrium [90Y] microsphere provided in the present application comprises a polystyrene sulfonic acid type resin microsphere and yttrium-90 adsorbed within the polystyrene sulfonic acid type resin microsphere. The use of polystyrene sulfonic acid type resin microsphere to adsorb yttrium-90 has significantly improved adsorption strength, significantly improved safety during preparation and use, and good killing effect on tumor cells.
2. The yttrium [90Y] microspheres provided in the present application can significantly improve the killing effect on liver cancer by controlling the particle size of polystyrene sulfonic acid resin microspheres to be in a range from 10 μm to 60 μm, preferably 28 μm to 43 μm, more preferably 30 μm to 35 μm.
The microsphere with a particle size 32.5 μm±2.5 μm has a density ranging from 1.125 g/ml to 1.6 g/ml in water (equivalent to red blood cells) and can withstand moist heat sterilization.
3. The yttrium [90Y] microspheres provided in the present application comprises an aqueous solution containing polyvinyl alcohol of 100 parts by volume; syrene of 2.5 to 5.5 parts by volume; divinylbenzene of 1.2 to 4.2 parts by volume; benzoyl peroxide of 0.02-2.02 parts by weight; toluene of 0.6 to 1.6 parts by volume; and n-heptane of 0 to 1.6 parts by volume. By controlling the ratio of various substances in the prescription within the optimal range, the polystyrene sulfonic acid type resin microspheres obtained have significantly increased ion capacity.
4. The yttrium [90Y] microsphere injection provided in the present application comprises the yttrium [90Y] microsphere in the present application. Polystyrene sulfonic acid type resin microsphere is used to adsorb yttrium 90, and less yttrium ions are precipitated. The safety during preparation and use is significantly improved, and the killing effect on tumor cells is good.
5. In the yttrium [90Y] microsphere injection provided in the present application, the resin microsphere is first washed with sodium phosphate solution, and then washed with sodium dihydrogen phosphate solution in the S2 step. The combination with yttrium ions is better, the precipitation of yttrium ions is less, and the safety is significantly improved.
The following embodiments are provided for a better understanding of the present invention but not for limiting the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by combining with the features of other prior art shall fall within the protection scope of the present invention.
If the specific experimental steps or conditions are not indicated in the examples, it can be carried out according to the operations or conditions of the conventional experimental steps described in the literature in this field. The reagents or instruments used without the manufacturer's indication are all conventional reagent products that can be obtained from commercial sources.
In the present application, liquid can be discharged through conventional commercially available filters, such as filters from manufacturers such as Merck Millipore or Steritech in the United States, model TKT04700/PCT12047100, polycarbonate, filter aperture 10 μm to 18 μm. The flow rate is 15-65 ml/min.
The process for preparing yttrium [90Y] microsphere injection comprises the following steps:
This example provides a method for preparing polystyrene sulfonic acid type resin microspheres, using the following reagents: polyvinyl alcohol (PVA), styrene (St), divinylbenzene (DVB), toluene, n-heptane, ethanol, and benzoyl peroxide (BPO).
The preparation method is as follows:
This example also provides an yttrium [90Y] microsphere injection and preparation method therefor. The prescription dosage of 10 bottles of injection was shown in Table 1:
Wherein, preparation of yttrium sulfate-89 solution: 0.3 g of yttrium oxide-89 powder was dissolved in 80 ml of 0.1 M sulfuric acid at a temperature of 40° C. to obtain an yttrium sulfate-89 solution, and place it in a production reserve container for future use.
The preparation method is as follows:
The radioactive activity of yttrium [90Y] microsphere injection was determined to be 6 GBq/bottle using the radioactive activity measurement method (General Rule 1401 of Part Four of the Pharmacopoeia of the People's Republic of China 2020 Edition).
This example provides a method for preparing polystyrene sulfonic acid type resin microspheres, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the collected polystyrene sulfonic acid type resin microspheres with a particle size ranging from 28 μm to 33 μm after sieving in Step S4.
This example provides an yttrium [90Y] microsphere injection and preparation method therefor, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the polystyrene sulfonic acid type resin microsphere prepared in this example was used to replace the polystyrene sulfonic acid type resin microsphere in Example 1.
This example provides a method for preparing polystyrene sulfonic acid type resin microspheres, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the collected polystyrene sulfonic acid type resin microspheres with a particle size ranging from 38 μm to 43 μm after sieving in Step S4.
This example provides an yttrium [90Y] microsphere injection and preparation method therefor, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the polystyrene sulfonic acid type resin microsphere prepared in this example was used to replace the polystyrene sulfonic acid type resin microsphere in Example 1.
This example provides a method for preparing polystyrene sulfonic acid type resin microspheres, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the collected polystyrene sulfonic acid type resin microspheres with a particle size ranging from 13 μm to 18 μm after sieving in Step S4.
This example provides an yttrium [90Y] microsphere injection and preparation method therefor, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the polystyrene sulfonic acid type resin microsphere prepared in this example was used to replace the polystyrene sulfonic acid type resin microsphere in Example 1.
This example provides a method for preparing polystyrene sulfonic acid type resin microspheres, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the collected polystyrene sulfonic acid type resin microspheres with a particle size ranging from 48 μm to 53 μm after sieving in Step S4.
This example provides an yttrium [90Y] microsphere injection and preparation method therefor, which is substantially the same as the raw materials and process of Example 1, with the only difference being that the polystyrene sulfonic acid type resin microsphere prepared in this example was used to replace the polystyrene sulfonic acid type resin microsphere in Example 1.
To investigate the effects of different pore forming agent and crosslinking agents during the preparation process of polystyrene sulfonic acid type resin microspheres as well as their amounts on the ion exchange capacity of polystyrene sulfonic acid resin microspheres, the preparation method was substantially the same as Example 1, with the only difference being the composition of the oil phase, as shown in the table below. The ion exchange capacity of polystyrene sulfonic acid resin microspheres prepared using the following groups A to C method and commercially available microspheres were tested.
The specific method was as follows: 100 ml of purified water was added to an appropriate amount of polystyrene sulfonic acid resin microspheres, stirred for 1 minute, washed repeatedly 3 times, and filtered. The filtered resin microspheres were weighted and recorded as M. 500 ml of 0.02M sodium hydroxide solution was added to the microspheres for immersion and then sonicated for 20 minutes. Filtered by suction, 100 ml of the filtrate was taken, two drops of phenolphthalein was added and then performed a titration experiment with 0.01M hydrochloric acid. The volume of hydrochloric acid used was recorded at the titration endpoint, recorded as V.
The calculation formula for ion exchange capacity was as follows: ion exchange capacity (mmol/g)=(molar concentration of sodium hydroxide solution×volume of sodium hydroxide-molar concentration of hydrochloric acid×V)/M.
From the above table, it can be seen that compared to commercially available products, the ion exchange capacity of the resin microspheres prepared in Groups A to C of the present application is significantly improved, especially group A, which used the same method as Example 1, showed significant improvement compared to both group B and group C.
Experimental nude mice weighing 20 g+2 g, all female, each was inoculated with 3×106 SMMC-7721 cells, when the tumor grew to 5 mm to 10 mm, they were randomly divided into experimental groups 1 to 5 and control group, with 5 mice in each group, and treatment was started. 1 mL of the injection prepared in Examples 1 to 5 were taken, diluted with water to a constant volume of 10 mL to obtain the test drug solutions for each group. Experimental groups 1 to 5 animals were injected intraperitoneally with the test drug solution (5 ml/animal) prepared in Examples 1 to 5, once daily for 10 consecutive days. The control group was given the same volume of water for injection and observed for 10 days. After 10 days of discontinuation, all animals were euthanized 24 hours after discontinuation, tumor masses were dissected, tumor weight was measured, and tumor inhibition rate was calculated according to the following formula:
From the above table, it can be seen that the microspheres prepared in Examples 1 to 3 has a much better killing effect on tumor cells than the microspheres prepared in Examples 4 and 5. Example 1 is the best, followed by Example 2.
To investigate the effects of using different phosphates as washing agent on the encapsulation efficiency yttrium [90Y] microspheres and dissolution rate of yttrium, the same method as Example 1 was used to prepare yttrium [90Y] microsphere injection. The difference only lies in the different types of phosphates in step (4) (in addition, the concentration of yttrium chloride [90Y] solution added to group D and group E is different). Group A all followed Example 1, that is, washing with sodium phosphate solution first and then washing with sodium dihydrogen phosphate solution, the specific operations of step (4) in Group A and other groups are shown in Table 5 below, and the other steps are the same as Example 1.
All the filtrate generated during the preparation process of yttrium [90Y] microsphere injection prepared by each group (including the steps of adding yttrium solution, first washing microspheres with water, washing microsphere with phosphate, and second washing with water) were collected and test solution 1 was obtained after 0.45 μm filtration. The concentration of yttrium ions in test solution 1 was measured by inductively coupled plasma mass spectrometer (recorded as yttrium ions in the filtrate).
The preservation solution of yttrium [90Y] microspheres prepared in each group was filtered out and test solution 2 was obtained after 0.45 μm filtration. The concentration of yttrium ions in test solution 2 was measured by inductively coupled plasma mass spectrometer (recorded as yttrium ions precipitated).
Preparation of standard curve: 50 μL of Y89 standard solution was transferred into a 10 ml volumetric flask, and volume with 2% nitric acid to obtain a 5 mg/L intermediate solution. 0.02 ml, 0.1 ml, 0.2 ml, 0.4 ml, and 1.0 ml of the intermediate solution were transferred to different 10 ml volumetric flasks, 20 μL Ge internal standard with a concentration of 10 mg/L was added, and volume with 2% nitric acid to obtain a series of standard solutions with Y89 concentrations of 10 μg/L, 50 μg/L, 100 μg/L, 200 μg/L, and 500 μg/L. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect standard solution, test solution 1, and test solution 2. The test conditions were as follows: collection mode: KED, RF power: 1550.0 W, cooling gas flow rate: 14.000 L/min, atomization gas flow rate: 1.0600 L/min, auxiliary gas flow rate: 0.8 L/min, collision gas flow rate: 4.340 ml/min. A standard curve was established based on the concentration of the test solution and the test results, and the concentration of yttrium ions in test solution 1 and test solution 2 was calculated based on the standard curve.
Due to previous experimental investigations, the methods of Group D and Group E were unable to adsorb a large amount of yttrium ions, resulting in a high concentration of yttrium ions in the filtrate and existing safety risk. Therefore, a 5 mg/mL yttrium chloride [90Y] solution was replaced with a 2 mg/ml yttrium chloride [90Y] solution and a 3.5 mg/mL yttrium chloride [90Y] solution to prepare yttrium [90Y] microsphere injection. From the experimental results, it can be seen that even if the concentration of yttrium ions added (i.e. the concentration of yttrium chloride [90Y] solution) was reduced, the concentration of yttrium ions and the concentration of precipitated yttrium ions in both groups of filtrates were still higher than the preferred solution A of the present application.
According to the above table, the amount of yttrium ions precipitation obtained in Group A experiment was significantly reduced, and no yttrium ions was detected in the filtrate. The resin microspheres prepared in Example 1 of the present application have good binding with yttrium ions, fewer yttrium ions precipitated, and significantly improved safety during preparation and use; however, other microspheres have poor binding with yttrium ions, and there are more yttrium ions in the filtrate.
Obviously, the above-mentioned embodiments are merely examples made for clear description, but do not limit the implementation. For those of ordinary skill in the art, other different forms of variations or modifications can also be made on the basis of the above-mentioned description. All embodiments are not necessary to be and cannot be exhaustively listed herein. In addition, the obvious variations or modifications derived therefrom all fall within the scope of protection of the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202311388190.4 | Oct 2023 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2023/128145 | 10/31/2023 | WO |
