CN108514645B

CN108514645B - Preparation with bone imaging and bone metastasis tumor treatment functions and preparation and application thereof

Info

Publication number: CN108514645B
Application number: CN201810306678.0A
Authority: CN
Inventors: 陈跃; 张蜀茂; 胥芹; 黄占文; 蔡亮; 张伟; 刘楠; 张莉
Original assignee: Affiliated Hospital of Southwest Medical University
Current assignee: Affiliated Hospital of Southwest Medical University
Priority date: 2018-04-08
Filing date: 2018-04-08
Publication date: 2021-07-20
Anticipated expiration: 2038-04-08
Also published as: CN108514645A

Abstract

The invention provides a preparation with bone imaging and bone metastasis tumor treatment functions, and belongs to the technical field of bone metastasis tumor treatment. The preparation is¹⁷⁷Lu-ibandronic acid. The invention also provides a preparation method of the preparation, and the preparation method adds ibandronic acid into the preparation¹⁷⁷LuCl₃In the solution, hydrochloric acid is used for adjusting the pH value of the solution, and the solution is sterilized and filtered after reacting for a certain time at a certain temperature. The invention relates to the use of radionuclides¹⁷⁷Lu and ibandronic acid are combined to provide a preparation which not only has bone imaging, but also can realize the treatment of bone metastasis¹⁷⁷Lu-ibandronic acid.¹⁷⁷Lu belongs to an osteogenic imaging agent, has a treatment effect on bone metastasis when used in a large dose, and has a strong effect on treating bone metastasis by using ibandronic acid¹⁷⁷The Lu-ibandronic acid preparation can better exert radionuclide¹⁷⁷Therapeutic effects of Lu and ibandronic acid on bone metastases.

Description

Preparation with bone imaging and bone metastasis tumor treatment functions and preparation and application thereof

Technical Field

The invention belongs to the technical field of bone metastasis tumor treatment, and particularly relates to a preparation with bone imaging and bone metastasis tumor treatment functions, and a preparation and an application thereof.

Background

The incidence of malignant tumors increases year by year, and bone is a common site of late metastasis of malignant tumors. Clinically, the treatment of bone metastasis is very tricky, and an effective and targeted treatment mode is urgently needed.

With the development of nuclear medicine, new radiopharmaceuticals and targeted drugs labeled with radiopharmaceuticals are continuously used for the therapeutic research of tumors, and the radionuclide therapy not only can play a role in relieving pain, but also can kill tumor cells, and even can enable part of patients to achieve a relieved state. The nuclides commonly used in China at present comprise phosphorus-32 (II) ((III))³²P), strontium-89 (⁸⁹Sr and samarium-153: (¹⁵³Sm), phosphorus-32 has relatively large toxic and side effects, has dose-limiting myelosuppression and is rarely used at present. In contrast to the 32P,⁸⁹sr can relieve bone pain to a certain extent, the application of Sr obviously reduces the use rate of analgesic drugs, and the combination with chemotherapeutic drugs has better curative effect, but the Sr has very low reaction section, very small production capacity and relatively high price, and can not be used for imaging. A member of the group Sm of the lanthanides,¹⁵³sm can be used for marking phosphate drugs, can be used for imaging and treating, and has better safety than the former two drugs, but¹⁵³Sm has a short half-life (1.95 days) and is not suitable for remote transport and the need for larger doses in batch processing also limits its use.

Disclosure of Invention

The invention aims to provide a preparation which has bone imaging and bone metastasis treatment effects, and a preparation method and application thereof. The purpose of the invention is realized by the following technical scheme:

a kind ofThe preparation has the functions of bone imaging and bone metastasis tumor treatment, and is¹⁷⁷Lu-ibandronic acid.

The invention also provides a preparation method of the preparation with the functions of bone imaging and bone metastasis tumor treatment, and the preparation method adds ibandronic acid into the preparation¹⁷⁷LuCl₃In the solution, hydrochloric acid is used for adjusting the pH value of the solution, and the solution is sterilized and filtered after reacting for a certain time at a certain temperature.

As a specific embodiment of the preparation method of the preparation with the functions of bone imaging and bone metastasis tumor treatment, the pH value of the hydrochloric acid adjusting solution is 4-5.

As a specific embodiment of the preparation method of the preparation with the functions of bone imaging and bone metastasis tumor treatment, the ibandronic acid is added in an amount of 1-1.5 mg, and the concentration is 25 mg/ml; the above-mentioned¹⁷⁷The activity of Lu is 0.25mCi, and the concentration is 10mCi/ml-20 mCi/ml.

As a specific embodiment of the preparation method of the preparation with the functions of bone imaging and bone metastasis tumor treatment, the reaction temperature is 90-100 ℃, and the reaction time is 1-1.5 hours.

As a specific example of the preparation method of the preparation with the functions of bone imaging and bone metastasis tumor treatment, the preparation method is that 1.5mg of ibandronic acid is added into the preparation¹⁷⁷Having a Lu activity of 0.25mCi¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 90 deg.C for 1 hr, sterilizing, and filtering.

The application of a preparation with the functions of bone imaging and bone metastasis tumor treatment, and the application of the preparation in the bone metastasis tumor treatment.

As a specific example of the application of the preparation with the functions of bone imaging and bone metastasis tumor treatment, the preparation is used as a radiopharmaceutical for targeted treatment of bone metastasis tumor.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a preparation with bone imaging and bone metastasis tumor treatment functions, and a preparation and an application thereof. The invention relates to the use of radionuclides¹⁷⁷Lu and ibandronic acid are combined to provide a preparation which not only has bone imaging, but also can realize the treatment of bone metastasis¹⁷⁷Lu-ibandronic acid.¹⁷⁷Lu belongs to an osteogenic imaging agent, has a treatment effect on bone metastasis when used in a large dose, and has a strong treatment effect on bone metastasis by using ibandronic acid¹⁷⁷The Lu-ibandronic acid preparation can better exert radionuclide¹⁷⁷Therapeutic effects of Lu and ibandronic acid on bone metastases.

Drawings

FIG. 1 shows a normal 2Kg rabbit injection¹⁷⁷SPECT/CT planar images of Lu-ibandronate at 4mCi were visualized at 1h, 2h, 12h, 24h, 48h and 6 d.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The preparation for treating both bone imaging and bone metastasis, the preparation and the application thereof according to the present invention will be described in detail below with reference to specific principles.

A preparation for simultaneously carrying out bone imaging and treating bone metastasis tumor is prepared¹⁷⁷Lu-ibandronic acid.

¹⁷⁷Lu is an ideal radionuclide, beta^-The particles have an average energy of 0.49MeV and are sufficiently low in energy to contain a high dose of low energy beta^-The radionuclide of the particle is lower than the equivalent dose and higher energy beta^-The particles are more effective, and the bone marrow damage degree caused by dosage superposition is more than that caused by other nuclides (¹⁵³Sm：E_βmax 0.81MeV，⁸⁹Sr：E_βmax 1.41MeV) is small, which makes it sufficiently safe, while gamma rays can be emitted: 113 (6.4%) and 208 (11.0%) keV, which can be used to monitor drug distribution and therapeutic effect in vivo, and which, due to their long half-life (6.73 days), are not only suitable for long-distance transport, but also reduce the radioactive safety problems associated with larger doses during batch processing。

In order to enable the radionuclide to have stronger targeting property and be selectively absorbed at the bone lesion part, the invention adopts ibandronic acid to carry out the radionuclide¹⁷⁷Lu for labeling. Ibandronic acid is one of the most powerful bisphosphonates at present, and the bisphosphonates contain a phospho-carbo-phospho core structure that facilitates their binding to mineralized bone matrix, leading to a high concentration of bisphosphonate drugs in the resorption cavity. Meanwhile, the medicine promotes osteoclast apoptosis, thereby reducing osteoclast number, inhibiting bone resorption, reversing the progress of osteolysis pathological changes, effectively relieving bone pain, reducing relapse to a certain extent and improving the survival rate of patients. Ibandronic acid can inhibit the activity of osteoclast and induce the apoptosis of osteoclast, has the capability of bone resorption resistance obviously higher than that of the first generation and the second generation of bisphosphonate, has long action time and small administration dosage, and can be widely used for treating hypercalcemia, bone metastasis and osteoporosis caused by tumor and preventing the complication of patients with bone metastasis. Ibandronic acid having the formula C₉H₂₃NO₇P₂Molecular weight of 319.22902, and structural formula as follows:

there are studies showing that ibandronic acid is less nephrotoxic in patients with multiple myeloma after treatment with zoledronic acid and ibandronic acid. Ibandronic acid can inhibit the increase of osteoclast activity and the release of bone calcium caused by various stimulating factors released by tumors, has few adverse reactions and light damage to renal function, and has the effects of preventing, treating and delaying the occurrence of bone-related events.

Therefore, the present invention relates to a radionuclide¹⁷⁷Lu and ibandronic acid are combined to provide a preparation which not only has bone imaging, but also can realize the treatment of bone metastasis¹⁷⁷Lu-ibandronic acid.¹⁷⁷Lu belongs to an osteogenic imaging agent, has a treatment effect on bone metastasis when used in a large dose, and has a strong treatment effect on bone metastasis by using ibandronic acid¹⁷⁷The Lu-ibandronic acid preparation can be better playedRadionuclide¹⁷⁷Therapeutic effects of Lu and ibandronic acid on bone metastases. Alone¹⁷⁷lu is an osteotropic nuclide that is taken up by the bone, but the targeting of treatment for bone metastases is not as strong as after ibandronic acid, which is not imaging. The invention relates to the use of radionuclides¹⁷⁷Lu and ibandronic acid are combined to achieve imaging, and the targeting of the preparation to the treatment of bone metastasis can be enhanced. The treatment effect on the specificity of the bone metastasis tumor is improved, and the effect that one plus one is greater than two can be achieved.

A preparation method of a preparation with bone imaging and bone metastasis tumor treatment functions comprises the step of adding ibandronic acid into the preparation¹⁷⁷LuCl₃In the solution, hydrochloric acid is used for adjusting the pH value of the solution, and the solution is sterilized and filtered after reaction at a certain temperature. The preparation method adopts phosphoric acid pairs¹⁷⁷LuCl₃Leaching to obtain radionuclide¹⁷⁷Lu is adjusted by hydrochloric acid to avoid bringing other impurities, and the Lu is prepared by the preparation method¹⁷⁷The radiochemical purity of the Lu-ibandronic acid product is not lower than 90-95 percent.

Further, the pH value of the hydrochloric acid adjusting solution is 4-5, and the pH value is adjusted to 4-5, so that the marking rate of the product can be guaranteed to be optimal.

Further, the adding amount of the ibandronic acid is 1-1.5 mg, and the concentration is 25 mg/ml; the above-mentioned¹⁷⁷The activity of Lu is 0.25mCi, and the concentration is 10mCi/ml-20 mCi/ml. The success rate of labeling is relatively higher at higher concentrations, but at too high concentrations small volumes of product are not preferred and the walls of the bottle remain too much to be wasteful, so that too high concentrations are not necessary.

Further, the reaction temperature is 90-100 ℃, and the reaction time is 1-1.5 h.

Further, the preparation method is that 1.5mg of ibandronic acid is added¹⁷⁷Having a Lu activity of 0.25mCi¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 90 deg.C for 1 hr, sterilizing, and filtering.

The preparation method is the invention¹⁷⁷Lu-ibandronic acid is preferredPreparation conditions, the preparation of which gives¹⁷⁷The labeling rate of the Lu-ibandronic acid product, namely the radiochemical purity can reach 97 percent.

The application of a preparation with the functions of bone imaging and bone metastasis tumor treatment, and the application of the preparation in the bone metastasis tumor treatment. Further, the formulation is useful as a radiopharmaceutical for targeted therapy of bone metastases.

In the formulation of the present invention, the active ingredient,¹⁷⁷lu belongs to an osteogenic imaging agent, has a treatment effect on bone metastasis when used in a large dose, has a strong treatment effect on bone metastasis by ibandronic acid, and is combined to obtain a preparation¹⁷⁷Lu-ibandronic acid can better exert radionuclide¹⁷⁷Lu and ibandronic acid have the treatment effect on bone metastasis tumor, and the effect that one plus one is larger than two is achieved.

The preparation, preparation and application of the preparation for bone imaging and bone metastasis therapy are further described in the following with reference to specific examples.

Example 1

This example¹⁷⁷The specific preparation method of Lu-ibandronic acid is as follows:

ibandronic acid 1.5mg was added¹⁷⁷Having a Lu activity of 0.25mCi (10mCi/ml)¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 90 deg.C for 1 hr, sterilizing with 0.22 μm sterilizing filter, and filtering. The labeling rate was determined by thin layer paper chromatography (TLC).

Prepared by the present example¹⁷⁷The radiochemical purity of the Lu-ibandronic acid product was 97%.

Example 2

ibandronic acid 5mg was added¹⁷⁷Having a Lu activity of 2.5mCi (50mCi/ml)¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 60 deg.C for 2 hr, sterilizing with 0.22 μm sterilizing filter, and filtering. The labeling rate was determined by thin layer paper chromatography (TLC).

Prepared by the present example¹⁷⁷Radiochemical purity of Lu-ibandronic acid productThe content was found to be 96.5%.

Example 3

Ibandronic acid 5mg was added¹⁷⁷Having a Lu activity of 2.5mCi (50mCi/ml)¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 60 deg.C for 6 hr, sterilizing with 0.22 μm sterilizing filter, and filtering. The labeling rate was determined by thin layer paper chromatography (TLC).

Prepared by the present example¹⁷⁷The radiochemical purity of the Lu-ibandronic acid product was 95.4%.

Although the ratios and reaction conditions of examples 2 and 3 achieved greater than 95% radiochemical purity, the required reactant amount was greater than that of the optimum conditions (example 1), the reaction time was longer, the radiochemical purity did not increase further with the increase in reaction time, and higher radiochemical purity was achieved at 2h of reaction.

Example 4

Ibandronic acid 1mg was added¹⁷⁷Having a Lu activity of 0.25mCi (50mCi/ml)¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 60 deg.C for 1 hr, sterilizing with 0.22 μm sterilizing filter, and filtering. The labeling rate was determined by thin layer paper chromatography (TLC).

Prepared by the present example¹⁷⁷The radiochemical purity of the Lu-ibandronic acid product is 60.4%.

In example 4, the reaction was not complete and the radiochemical purity was low due to the insufficient reaction caused by the decrease in the amount of iban phosphate under the same conditions as in example 1.

In vitro stability test

Freshly prepared as in example 1¹⁷⁷Lu-ibandronic acid was incubated directly in a thermostatic water bath at room temperature and 37 ℃ and the radiochemical purity of the labeled compound was checked by TLC 2, 8, 12, 24, 48 and 72h after labeling, respectively. Another 0.1ml of fresh human serum was diluted 10-fold and freshly prepared as in example 1 was added¹⁷⁷Lu-ibandronic acid was left standing at room temperature and 37 ℃ and the radiochemical purity of the labeled compound was checked by TLC at 2, 8, 12, 24, 48 and 72h after labeling, respectively, and the results are shown in Table 1 below.

TABLE 1 results of radiochemical purity of the product of example 1 for different treatment regimes and treatment times

2h

8h

12h

24h

48h

72h

At normal temperature

97％

96％

95％

37℃

97％

96.5％

96％

Normal temperature (serum)

97％

96％

95％

37 ℃ (serum)

97％

96％

95.5％

As can be seen from table 1, it is,¹⁷⁷lu-ibandronic acid has better in vitro stability, and the radiochemical purity of the Lu-ibandronic acid is still 95 percent after the Lu-ibandronic acid is placed at room temperature for 72 hours.

Plasma protein binding rate

Fresh heparin anticoagulated human plasma 2mL is taken for standby. 3 5mL tubes were prepared and 0.1mL of human plasma 0.1m L was added to each tube, followed by 0.1mL of the freshly prepared sample of example 1, about 37KBq¹⁷⁷Lu-ibandronic acid solution, which was then incubated in an incubator at 37 ℃ for 2 h. Adding 1mL of 25% trichloroacetic acid solution into each test tube, uniformly mixing by using a vortex mixer, centrifuging at 2000r/min for 5min, and collecting supernatant; adding 1mL of 10% trichloroacetic acid solution into the precipitate, uniformly mixing by using a vortex mixer, centrifuging at 2000r/min for 5min, and collecting supernatant; repeat 3 times. A gamma counter measures total radioactive Counts (CPM) of the pellet and supernatant, respectively, and calculates the plasma protein binding rate (PPB).

The results show that example 1 produces¹⁷⁷The plasma protein binding rate (PPB) of the Lu-ibandronic acid solution was 74.14%. The higher the plasma protein binding rate of the marker, the longer the residence time in the body is, and the less rapidly the marker is removed. Example 1 preparation of¹⁷⁷Lu-The plasma protein binding rate of the ibandronic acid solution is 74.14 percent, the ibandronic acid solution stays in a body for a long time and is not easy to remove, and a better treatment effect can be achieved.

Coefficient of distribution of fat and water

3 tubes were taken and 1ml of saturated n-octanol and 0.5ml of deionized water were added to each tube, followed by 0.01mCi of freshly prepared as in example 1¹⁷⁷Lu-ibandronic acid is mixed and vibrated for 20min by using a vortex mixer, centrifuged for 5min at 2000r/min, 6 test tubes are taken and divided into A, B groups, the numbers of the test tubes are respectively A-1, B-1, A-2, B-2, A-3 and B-3, after two-phase separation, the group A correspondingly collects 0.1ml of organic phase, the group B correspondingly collects 0.1ml of water phase, the radioactivity counts of the organic phase and the water phase are respectively measured, and the lipid-water distribution coefficient is calculated.

The results show that example 1 produces¹⁷⁷The lipid-water distribution coefficient (Pc) of the Lu-ibandronic acid solution was 0.001476 at pH 7.

¹⁷⁷Lu-ibandronic acid distributed in mice

20 healthy Kunming mice (body weight 18-22 g) are divided into 5 groups at random and injected into the tail vein of the mice prepared in example 1¹⁷⁷Lu-ibandronic acid 3.7MBq/0.1ml, each of 4 cases after 1h, 4h, 24h, 48h and 6 days was decapitated and sacrificed, blood, heart, lung, liver, spleen, kidney, stomach, intestine, muscle and bone tissues were weighed and measured for radioactivity, and the injection dose per gram of tissue (% ID/g) was calculated.

¹⁷⁷The distribution results of Lu-ibandronic acid in mice are shown in Table 2 below:

TABLE 2¹⁷⁷Distribution of Lu-ibandronic acid in mice

As can be seen from Table 2 above, the bone pairs¹⁷⁷Lu-ibandronic acid hasThe high intake is 1h after injection, the bone intake is higher than other organs, and reaches the maximum value at 48h, and then the intake in the bone is reduced along with the prolonging of time, until the 6 th day, the ratio of the bone to other organs is still higher, which shows that 177 Lu-ibandronic acid stays on the bone for a longer time, and a better treatment effect can be achieved.

¹⁷⁷Imaging of Lu-ibandronic acid at different time points in rabbits

1Kg of rabbit was injected into the ear vein of 1 rabbit¹⁷⁷Lu-ibandronic acid 4mCi, for planar imaging at 1h, 2h, 12h, 24h, 48h and 6d, respectively.¹⁷⁷The results of Lu-ibandronic acid imaging at different time points in normal rabbits are shown in FIG. 1.

FIG. 1 shows normal rabbit injections¹⁷⁷SPECT/CT planar images of Lu-ibandronate at 4mCi were visualized at 1h, 2h, 12h, 24h, 48h and 6 d. As can be seen in FIG. 1, the injection was carried out¹⁷⁷Lu-ibandronic acid starts to have light images of the spine, ribs and bones of limbs after 1h, the two kidneys are light, the bladder is thick and the images are visible¹⁷⁷Lu-ibandronic acid is excreted via the urinary system. At 12h, background is basically cleared, at this time, the imaging of the spine, ribs and limbs is clear, at 6 th day, the imaging agent is still taken into the spine, ribs and limbs, the contrast between bones and background is still high, and the contrast is visible¹⁷⁷Lu-ibandronic acid is an imaging agent which has high bone uptake, quick soft tissue clearance, low liver and spleen uptake and is excreted by the urinary system, has long retention time on bones, and can achieve better treatment effect when being applied to patients with bone metastasis.

¹⁷⁷Lu-ibandronic acid and radionuclides¹⁷⁷Comparison of Lu Effect

In order to verify the invention¹⁷⁷The combination of Lu and ibandronic acid has good therapeutic effect on bone metastasis, and the inventor prepares the compound obtained in example 1¹⁷⁷Lu-ibandronic acid and individual radionuclides¹⁷⁷Lu was injected into mice separately (injection amount is equivalent, concrete operation mode and above example)¹⁷⁷Lu-ibandronic acid was distributed equally in the mice) and its distribution in different organs of the mice at different times was observed.¹⁷⁷The distribution of Lu-ibandronic acid in mice at different times is shown in Table 2 above, with the individual radionuclides¹⁷⁷The distribution of Lu in mice at different times is shown in Table 3 below.

TABLE 3¹⁷⁷Distribution of Lu in mice

From the data results and comparative analysis of tables 2 and 3 above, the following conclusions can be drawn:

1、¹⁷⁷lu and¹⁷⁷both Lu-ibandronic acid (IBN) were distributed highest in the bone and peaked at 48 h.

2、¹⁷⁷The bone uptake of Lu-IBN mice was higher than at 1h later¹⁷⁷The uptake of Lu, Explanation¹⁷⁷Lu-IBN is relatively high¹⁷⁷Lu is better taken up by bone tissue.

3. Compared with the prior art, the utility model has the advantages that,¹⁷⁷the uptake of Lu-IBN in other organs is significantly lower than that of Lu-IBN¹⁷⁷Lu, description¹⁷⁷Lu-IBN is relatively high¹⁷⁷Lu has a higher bone/organ ratio. Higher bone/kidney ratio, indicates¹⁷⁷Lu-IBN is excreted more rapidly through renal metabolism; higher bone/muscle ratio, lower soft tissue background, and is more beneficial to bone imaging. Higher bone/liver ratio, then¹⁷⁷Liver imaging by Lu-IBN will be lower than¹⁷⁷Lu; and so on.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. A preparation for simultaneously performing bone imaging and bone metastasis tumor treatment is characterized by being¹⁷⁷Lu-ibandronic acid; the preparation method of the preparationBy adding ibandronic acid¹⁷⁷LuCl₃And (3) in the solution, adjusting the pH value of the solution by using hydrochloric acid, reacting at the temperature of 90-100 ℃ for 1-1.5 h, sterilizing and filtering.

2. The formulation of claim 1, wherein the hydrochloric acid is used to adjust the pH of the solution to 4-5.

3. The preparation for treating both bone imaging and bone metastasis tumor according to claim 1, wherein ibandronic acid is added in an amount of 1-1.5 mg at a concentration of 25 mg/ml; the above-mentioned¹⁷⁷The activity of Lu is 0.25mCi, and the concentration is 10mCi/ml-20 mCi/ml.

4. The formulation of claim 1, wherein the formulation is prepared by adding 1.5mg ibandronic acid to the formulation for both bone imaging and bone metastasis therapy¹⁷⁷Having a Lu activity of 0.25mCi¹⁷⁷LuCl₃Adjusting pH of the solution to 5 with hydrochloric acid, reacting at 90 deg.C for 1 hr, sterilizing, and filtering.

5. The use of the formulation for both bone imaging and bone metastasis therapy according to claim 1, wherein said formulation is used in the preparation of a product for bone metastasis therapy.

6. The use of the formulation for both bone imaging and bone metastasis therapy according to claim 5, wherein said formulation is used in the preparation of a radioactive product for targeted therapy of bone metastasis.