CN1861616A - Compound for tumour display and caring and preparation process thereof - Google Patents

Abstract

The invention discloses a radioactive rhenium-labeled diethylenetriaminepentaacetic acid-deoxyglucose (DTPA-DG) for diagnosis and treatment with high labeling rate and good stability and a preparation method thereof. The preparation method of the present invention is characterized in that an appropriate amount of reaction ligand, radioactive rhenium, sodium gluconate solution and reducing agent are added into a reactor, and the reaction system is adjusted for a period of time to obtain the desired product. Radioactive rhenium-DTPA-DG is expected to be used in the radiological diagnosis and treatment of solid tumors. The preparation prepared by the present invention, coupled with a ¹⁸⁸ W- ¹⁸⁸ Re generator, can prepare ¹⁸⁸ Re-labeled DTPA-DG preparations at any time, and has the advantages of simple method, easy operation, high tumor uptake, good retention, high target/non-target ratio and low price Advantages, easy to use in medical institutions, and more convenient to use in units without cyclotrons.

Description

Complex for tumor imaging and treatment and preparation method thereof

technical field

The invention belongs to the field of medical compounds, in particular to an acyl sugar derivative of radioactive rhenium-labeled diethylenetriaminepentaacetic acid (DTPA) and a preparation method thereof.

Background technique

Cancer is a common disease that seriously endangers health. Cancer has become the first cause of death. Cancer prevention and research have become a topic of concern to scientists all over the world.

The glycolysis of malignant tumors is enhanced, and the glucose analogue 18F-FDG (18F-fluorodeoxyglucose) can be taken up by tumor cells, and the level of 18F-FDG in tumor tissues is significantly higher than that in surrounding normal tissues and benign lesions. 18F-FDG PET (Positron Emission Computed Tomography) non-invasively detects tumor glucose metabolism, plays an important role in the early diagnosis of malignant tumors, formulates tumor treatment, curative effect judgment, and prognosis evaluation for clinicians. 18F-FDG drugs need to be produced by cyclotron, which is very expensive; the half-life is short, only 110min; the PET imaging equipment used is expensive; and the production time of drug synthesis is long. 18F-FDG PET tumor imaging has been clinically used for more than 20 years, but its popularization and application in hospitals at all levels is greatly limited due to its high cost and expense.

Later, people found that 99mTc has ideal nuclear physical properties, its half-life is 6.02h, emits 140keV gamma rays, and 99mTc generators are easy to obtain. The half-life, energy and rays of 99mTc are the most ideal gamma imaging radionuclides. If 99mTc can be used to label glucose analogues, it will play an important clinical role and is expected to be widely used in basic medical research and clinical practice. It has become a hot spot in the development of nuclide imaging agents. In 2003, Yang DJ first reported the synthesis of dicysteine (EC)-DG and labeled it with 99mTc; the distribution study of 99mTc-ECDG showed that the ratio of tumor/brain and tumor/muscle was higher than that of 18F-FDG, and 99mTc-ECDG imaging can be clear Shows malignant tumor foci. However, the distribution of 99mTc-ECDG showed that the tumor/blood and tumor/lung ratios were less than 1.0, which was lower than that of 18F-FDG; 99mTc-ECDG was not yet an ideal 99mTc-labeled DG imaging agent, and there was a large gap with 18F-FDG.

The inventors completed 99mTc labeling of acyl sugar derivatives of diethylenetriaminepentaacetic acid (DTPA), which has high labeling rate, good stability in vitro, easy operation, early non-invasive diagnosis of tumors, and low imaging cost (see Chinese patent application 200510020334.6 ). However, 99mTc-labeled DG imaging agent only emits γ particles and cannot be used for tumor treatment.

Contents of the invention

The object of the present invention is to overcome the disadvantages of the above-mentioned prior art and provide a radioactive rhenium-labeled complex with excellent therapeutic and imaging properties and good stability in vitro.

Another object of the present invention is to provide a new preparation method of radioactive rhenium-labeled complex, which is simple in preparation method, simple in labeling method and high in labeling rate.

The inventor considers that ¹⁸⁶ Re or ¹⁸⁸ Re, like 99mTc, can emit gamma particles suitable for imaging, but unlike 99mTc, they can also emit beta particles, which have moderate energy and can be used for treatment at the same time, ¹⁸⁶ Both Re and ¹⁸⁸ Re have longer half-lives, and ¹⁸⁸ Re can be provided by the generator, which is convenient to use and is an ideal therapeutic nuclide. However, it is necessary to find a complex compound suitable for forming a complex with radioactive rhenium before using radioactive The rhenium-labeled complex is labeled with deoxyglucose, and then used for tumor targeting therapy and radiographic imaging research.

In order to apply the above theory in practice, on the basis that the inventor has completed the following ligand compounds, he proposed the technical idea of using radioactive rhenium-labeled diethylenetriaminepentaacetic acid (DTPA) acyl sugar derivatives, To find a complex that can achieve the purpose of the invention:

The ligand compound used for imaging diagnosis is an acyl sugar derivative of diethylenetriaminepentaacetic acid (DTPA), and its structure is shown in the general formula (II)

_R in the structure of formula (II) is five-carbon sugar, six-carbon sugar, polysaccharide, and mercapto, amino, amino, cyano, carboxyl or ester group substituted deoxypentose, deoxyhexose, deoxypolysaccharide structure , R ₂ and R ₃ are H respectively, or five-carbon sugar, six-carbon sugar, polysaccharide, and deoxy-pentose, deoxy-hexose, deoxygenated sulfhydryl, amino, amino, cyano, carboxyl or ester polysaccharide structure.

After countless experiments, the radioactive rhenium was combined with the above-mentioned complex compound, and the complex with the following general structure was obtained:

In formula (I), a is 186 or 188, R ₁ is sugar or sugar amine, R ₂ and R ₃ are H and/or sugar or sugar amine respectively.

The complex is prepared as follows:

Add reaction ligand, weak complexing agent, radioactive rhenium and reducing agent into a closed reaction container; adjust the pH of the reaction system between 2-5.5; heat in a water bath to keep the reaction temperature at 20-100°C, and place it for 30-180 minutes; The reaction ligand is any one of DTPA-DG, DTPA-2DG, DTPA-3DG, the weak complexing agent used is sodium gluconate, and the reducing agent used is SnCl ₂ ·2H ₂ O solution saturated with inert gas; Under the protection of inert gas.

Wherein, the dosage of reducing agent SnCl ₂ ·2H ₂ O solution is 0.5-10mg/ml, and its inert gas is nitrogen;

The inert gas used for reaction protection is any one of helium, neon, argon and nitrogen.

It should be noted that the weak complexing agent is sodium gluconate, which is an intermediate ligand.

The preparation process of the above-mentioned complex generally can adopt:

1. Preparation of acyl sugar derivatives of diethylenetriaminepentaacetic acid

Under stirring, dihalosulfoxide (such as the most commonly used SOCl ₂ ) is dropped into diethylenetriaminepentaacetic acid (DTPA) at -30°C-0°C, stirred at -30°C-100°C, and the reactant is refluxed for 12 -28 hours, steam the excess dihalide sulfoxide under reduced pressure to obtain the acid halide bisanhydride intermediate compound, which can be directly carried out in the next step without separation; in the obtained acid halide bisanhydride, add DMSO, pyridine and the corresponding sugar or Substitute sugar (such as 2-amino D-glucose hydrochloride, etc.), and stir the mixture at -50°C-196°C for 24-48 hours. The reaction solution after the reaction is separated and purified by means of dialysis membrane or Sephadex (G10; G15; or G25) to obtain the corresponding structural product.

2. Preparation of complexes

Medical radioactive rhenium complex of the present invention, its general chemical structure formula (I) is as follows:

In formula (I), a is 186 or 188, R ₁ is sugar or sugar amine, R ₂ and R ₃ are H and/or sugar or sugar amine respectively.

The medical radioactive rhenium complex of the present invention is prepared by reacting radioactive perrhenate (ReO ₄ ^- ) with acyl sugar derivatives of diethylenetriaminepentaacetic acid in the presence of a reducing agent to obtain the radioactive rhenium-labeled diethylenetri Acyl sugar derivatives of aminopentaacetic acid.

The ligand in the reaction system can be any one of DTPA-DG, DTPA-2DG, and DTPA-3DG.

Rhenium exists in the form of perrhenate (ReO ₄ ^- in solution), and rhenium is ¹⁸⁶ Re or ¹⁸⁸ Re.

Weak complexing agent sodium gluconate solution as an intermediate ligand, on the one hand, stabilizes the effect of Sn ²⁺ and reduces the hydrolysis of Sn ²⁺ ; on the other hand, it can maintain weak complexation with ¹⁸⁸ Re, so that ¹⁸⁸ Re is in a stable and low price state , to ensure the smooth progress of the substitution reaction.

In order to prevent the reaction product from being oxidized, the reaction is carried out under the protection of helium, neon, argon, nitrogen, and nitrogen is usually used.

The reducing agent is tin dichloride, and the dosage of SnCl ₂ ·2H ₂ O is 0.5-10 mg.

The pH value of the reaction system is adjusted between 2-5.5 with common acid and base.

The suitable reaction temperature is 20-100°C.

Symbol Description:

Re: Represents the element rhenium, with or without radioactive isotopes.

¹⁸⁶ Re and ¹⁸⁸ Re: two radioactive isotopes of rhenium with atomic weights 186 and 188, respectively.

DTPA-DG: stands for diethylenetriaminepentaacetic acid-2-glucosamine (deoxyglucose) or its salt.

¹⁸⁶ Re or ¹⁸⁸ Re-DTPA-DG: represent rhenium-186 or rhenium-188 diethylenetriaminepentaacetic acid-glucosamine (deoxyglucose) or its salts, respectively.

3. The treatment and imaging experiments of radioactive rhenium-labeled complexes and the comparison results of ^99m Tc-DTPA-DG:

The biodistribution of ^99m Tc-DTPA-DG breast cancer MCF-7 nude mice showed that the tumor/blood ratios were 1.29 and 3.13 at 1h and 2h respectively, and the tumor/muscle ratios were 2.63 and 5.01 at 1h and 2h respectively. 12h after intravenous injection of ¹⁸⁸ Re-DTPA-DG in nude mice with breast cancer MCF-7, the content ratio of the tumor to the surrounding muscle was as high as 12.76.

^{Imaging of 99m} Tc-DTPA-DG in nude mice bearing MCF-7 breast cancer showed high uptake in tumor tissue, and T/NT values were 2.46 and 3.54 at 0.5h and 2h, respectively. Imaging of ¹⁸⁸ Re-DTPA-DG-bearing MCF-7 breast cancer nude mice showed high tumor tissue uptake, and T/NT values were 5.9 and 7.8 at 12h and 24h, respectively.

In vivo biodistribution and imaging of ¹⁸⁸ Re-DTPA-DG tumor-bearing mice showed higher tumor uptake, higher radioactivity in tumor than in blood, lung, muscle, brain tissue, tumor/blood, tumor/lung, tumor/ Muscle and tumor/brain ratios were high.

After 3 weeks of tail vein injection of ¹⁸⁸ Re-DTPA-DG in tumor-bearing mice, the tumor inhibition rate was obvious, and the tumor volume was significantly lower than that of perrhenate eluate group and normal saline group.

The radioactive rhenium-labeled acyl sugar derivative of diethylenetriaminepentaacetic acid provided by the invention has a simple labeling method, is easy to prepare, has high labeling rate and low cost. Biological experiment results show that it has high uptake and good retention in tumor tissue, has a high target/non-target ratio, and is suitable for use as a tumor therapeutic agent and an imaging agent.

Description of drawings

Figure 1 is an anterior plane image of breast cancer MCF-7 nude mice 12 hours after injection of ¹⁸⁸ Re-DTPA-DG complex;

Fig. 2 is an anterior plane image of breast cancer MCF-7 nude mice 24 hours after injection of ¹⁸⁸ Re-DTPA-DG complex.

Detailed ways

Below in conjunction with accompanying drawing and embodiment the present invention is described in further detail, but the present invention is not limited to these examples:

Example 1:

1) Add 10ml of dihalosulfoxide (SOCl ₂ ) dropwise into 3.94g of diethylenetriaminepentaacetic acid (DTPA) (IV) at -30°C under stirring, and continue stirring at room temperature for 3 hours. The test results show that the temperature range of -30°C to 100°C in this process generally does not show significant adverse effects on the results. Then the reactant was refluxed for 20 hours, and excess SOCl ₂ was distilled off under reduced pressure to obtain 3.76 g of light yellow powder of the intermediate compound of acid chloride dianhydride, which can be directly carried out to the next reaction without separation.

2) In the acid chloride bisanhydride 3.76g (0.010mol) obtained in the previous step, add 30ml DMSO, 5ml pyridine and 2.85g (0.0132mol) 2-amino-D-glucose hydrochloride, the mixture can generally be stirred at room temperature for 24 hours (the operating temperature of this process can be allowed to be 20°C-150°C) to obtain a transparent brown solution. The solution is separated and purified by dialysis membrane or Sephadex (G10) to obtain the condensate of DTPA and D-glucosamine (DTPA-DG) represented by formula (II).

MS (m/e): 555 (M ⁺ ), 394, 351

¹ H-NMR (ppm): δ12.2(4H,b); δ8.1(b); δ5.2(1H); δ4.6(1H,); δ3.8(2H); 2H); δ3.6(1H); δ3.5(10H); δ3.1(4H); δ2.9(4H); δ2.5(4H)

3) Preparation of ¹⁸⁸ Re-DTPA-DG complex:

Take DTPA-DG 10 mg, dissolve it in a test tube of 400 μL pH value 5.5 phosphate buffer solution, wait for complete dissolution, add 100 μL of 0.5mol/L sodium gluconate solution, add radioactive perrhenate ¹⁸⁸ ReO ₄ Na 100-200 μL, mix well , the solution is transparent, add SnCl ₂ ·2H ₂ O 4mg, adjust the pH of the reaction system to 5.5, shake and mix immediately, fill with nitrogen and seal, place in a 38°C incubator, react for 3 hours, and mark is complete.

The radiochemical purity of the ¹⁸⁸ Re complex was determined by TLC. TLC was carried out on 10cm Xinhua No. 1 chromatography paper. Place 1-2 μL sample at the starting point of the chromatographic strip, develop it with 0.9% NaCl and acetone solution respectively, after development, let it dry naturally in the air, divide it into 11 sections, and measure the radioactive count of each section. For normal saline upward expansion, the ratio shift value Rf of ¹⁸⁸ Re-DTPA-DG and ReO ₄ ^- is 0.9-1.0, and the Rf of ReO ₂ is 0; for upward expansion of acetone, ¹⁸⁶ Re (or ¹⁸⁸ Re)-DTPA-DG and ReO ₂ The Rf of ReO 4 - is 0, and the Rf of ReO ₄ ^- is 1.0.

Stability of the ¹⁸⁸ Re complex of DTPA-DG:

^{The 188} Re-DTPA-DG prepared according to Example 1 was placed at room temperature, sampled for 1h, 2h, 3h, 6h, and 12h respectively, and analyzed by TLC. The Xinhua No. 1 chromatography paper was used as a support, and 0.9% The radiochemical purity of ⁹⁹ Tc ^m -DTPA-DG was determined using NaCl and acetone solution as developing solvents. The marker ¹⁸⁸ Re-DTPA-DG has good stability, and the radiochemical purity still reaches 89.2% after being placed at room temperature for 12 hours.

The performance measurement of ¹⁸⁸ Re-DTPA-DG is described as follows:

1. Biodistribution of ¹⁸⁸ Re-DTPA-DG breast cancer MCF-7 nude mice:

Select female nude mice with a body weight of about 25 g, and subcutaneously inoculate the human breast cancer MCF- ⁷ cell line in the left thigh. 1.0cm is used for experiments. Nine nude mice with breast cancer MCF-7 were divided into 3 groups, 3 in each group, injected with Re-DTPA-DG3.7MBq/0.1ml through the tail vein, the nude mice were decapitated 3h, 12h, and 24h after injection, and the blood, Heart, lung, liver, spleen, kidney, stomach, small intestine, muscle, tumor and other tissues and organs, weigh and measure their radioactive counts, calculate the percent injection dose per gram of tissue (%ID/g), and calculate the organ/blood ratio and tumor/organ (T/NT) ratio.

See Table 1 and Table 2 for the in vivo distribution test results of ¹⁸⁸ Re-DTPA-DG in tumor-bearing mice. The table shows that ¹⁸⁸ Re-DTPA-DG showed higher tumor uptake when the injection time was 3, 12, and 24 hours respectively. The radioactivity of the tumor at 3, 12, and 24 hours accounted for 1.98±0.29ID%·g ^-1 , 2.89±0.43ID%·g ^-1 , 0.42±0.06ID%·g ^-1 of the injected dose, respectively. ¹⁸⁸ Re-DTPA-DG has high radioactivity in kidney, no obvious radioactivity distribution in stomach, low intestinal radioactivity, radiopharmaceuticals are stable in vivo, and the formation of free ReO ₄ ^- is rare. Radiotracers are mainly excreted by the kidneys and into the bladder through urine. The distribution of radioactivity in brain tissue is low. The distribution of radioactivity in brain tissue is low. Because DTPA-DG is water-soluble, its molecular weight is larger than that of FDG, and it is difficult to pass through the blood-brain barrier.

The radioactivity in the tumor is higher than that in blood, lung, muscle, and brain tissue. At 12 hours after injection, the ratios of tumor/blood, tumor/lung, tumor/muscle, and tumor/brain are 11.00, 5.22, 12.76, and 16.72, respectively. The tumor within 24 hours / blood ratio, tumor / muscle ratio were high. The uptake of ¹⁸⁸ Re-DTPA-DG in tumor tissue is high, which is feasible for tumor treatment and imaging research.

Table 1 Tissue distribution of ¹⁸⁸ Re-DTPA-DG in tumor-bearing nude mice (x±s)ID%·g ^-1 organ t/h 3 12 twenty four Blood 0.40±0.06 0.26±0.04 0.05±0.01 heart 0.81±0.12 0.42±0.06 0.21±0.03 lung 0.35±0.05 0.55±0.08 0.19±0.03 liver 1.26±0.18 0.53±0.08 0.30±0.04 spleen 0.34±0.08 0.21±0.03 0.37±0.06 kidney 1.94±0.25 2.10±0.32 0.15±0.02 Stomach 0.76±0.11 0.51±0.08 0.32±0.05 small intestine 0.47±0.07 0.30±0.05 0.36±0.05 muscle 0.27±0.04 0.23±0.03 0.12±0.02 brain 0.11±0.02 0.17±0.03 0.04±0.01 the tumor 1.98±0.29 2.89±0.43 0.42±0.06

Table 2 T/NT values of ¹⁸⁸ Re-DTPA-DG tumor-bearing nude mice T/NT t/h 3 12 twenty four tumor/blood 4.99 11.00 8.20 tumor/lung 5.68 5.22 2.18 tumor/muscle 7.40 12.76 3.56 tumor/brain 18.16 16.72 10.98

2. Imaging experiment of ¹⁸⁸ Re-DTPA-DG breast cancer MCF-7 nude mice:

Select female nude mice with a body weight of about 25 g, and subcutaneously inoculate the human breast cancer MCF-7 cell line in the left thigh. The cell dosage is 0.2 mL, containing about 1× ¹⁰⁷ tumor cells. 1.0cm is used for experiments. Three nude mice with breast cancer MCF-7 were injected with ¹⁸⁸ Re-DTPA-DG 18.5MBq/0.1mL through the tail vein, and collected statically at 1, 2, 4, 12, and 24 hours after injection to observe the effect ^{of 188} Re-DTPA-DG on the tumor According to the concentration of the lesion, the ratio of radioactive counts (T/B) between the lesion and the corresponding position of the uninjured thigh was calculated using the ROI technique.

¹⁸⁸ Re-DTPA-DG breast cancer MCF-7 nude mouse imaging From Figure 1 and Figure 2, it can be seen that the tumor focus is clearly displayed, and its radioactivity is higher than that of the contralateral side by naked eye observation. Kidney shadows were seen, bladder radioactivity gradually increased, and other organs such as brain, lung, small intestine, and muscle had no obvious high radioactivity distribution, and no thyroid and stomach development were seen. The 12h and 24h T/B ratios measured by ROI technique were 5.9 and 7.8 respectively. ¹⁸⁸ Re-DTPA-DG imaging of tumor-bearing animals can clearly show the tumor focus. ¹⁸⁸ Re-DTPA-DG had higher radioactivity in kidney, no obvious radioactivity distribution in stomach, and lower intestinal radioactivity. Radiotracers are mainly excreted by the kidneys and into the bladder through urine. ¹⁸⁸ Re-DTPA-DG imaging of tumor-bearing animals can clearly display tumor foci, which can be used for tumor diagnosis.

3. MCF-7 breast cancer cells uptake ¹⁸⁸ Re-DTPA-DG experiment:

Take 30 disposable test tubes, add 100μl cell suspension to each tube, set up ¹⁸⁸ Re-DTPA-DG group and ReO ₄ ^-group , add ¹⁸⁸ Re-DTPA-DG and ReO ₄ ^- eluent 3.7kBq/10μL respectively at the beginning of the experiment , placed in a 37 °C 5% CO ₂ incubator. After 0.5h, 1h, 2h, 3h, and 4h, centrifuge (1000 rpm) for 5 minutes, carefully absorb the supernatant with a disposable cotton swab, wash twice with PBS, centrifuge, discard the supernatant, SN-695B The type intelligent RIA gamma measuring instrument detects and counts cpm, and measures the total T tube count cpm. Calculate the tumor cell uptake rate of the experimental group and the control group at different time periods, the tumor cell uptake rate=(count per test tube/total T tube count)×100%, and the value is represented by x±s.

Table 3 shows the results of the uptake of ¹⁸⁸ Re-DTPA-DG by MCF-7 breast cancer cells. ¹⁸⁸ There were significant differences between Re-DTPA-DG group and ReO ₄ ^-group at different time periods (P<0.05=, the control group basically did not ingest, and the uptake rate of the experimental group gradually increased with time, and there were differences. Tumor cells The uptake of ¹⁸⁸ Re-DTPA-DG is high, and ¹⁸⁸ Re-DTPA-DG can enter cells for diagnosis and treatment research.

Table 3 Uptake rate of tumor cells (x±s) Tumor cell uptake rate (%) 0.5h 1h 2 hours 3 hours 4 hours ¹⁸⁸ Re-DTPA-DG group 9±0.91 10.8±1.73 11.8±1.08 14.1±0.17 15.6±0.49 ReO ₄ ^-group 2.1±0.2 2.4±0.31 2.6±0.39 3.4±0.54 3.4±0.21

4. ¹⁸⁸ Re-DTPA-DG tumor-bearing nude mice growth inhibition experiment:

Six nude mice were inoculated subcutaneously with MCF-7 breast cancer cells at 1×10 ⁷ mL ^-1 in the right forelimb, and the experiment began when the diameter of the tumor was about 10 mm. They were divided into normal saline control group, ReO ₄ ^-group and ¹⁸⁸ Re-DTPA-DG group. The normal saline control group was given 0.1 mL of normal saline, while the ReO ₄ ^- group and ^{the 188} Re-DTPA-DG group were given tail vein specific activity of 92.5 GBq/L (0.1 mL). The size of the tumor was observed every 2-3 days, and the volume change of the tumor was continuously observed for 3 weeks. Tumor inhibition rate=(1-experimental group volume/control group volume)×100%

¹⁸⁸ The tumor volume in the Re-DTPA-DG group after 21 days of treatment was significantly smaller than that in the ReO ₄ ^- group and the normal saline group, 823.6±50.58, 1153.0±99.42, and 1162.7±73.08mm ³ , respectively. There were significant differences among the ¹⁸⁸ Re-DTPA-DG group, the perrhenate eluate group and the normal saline group (F=13.893, P<0.01) (Table 4). 3 weeks after ¹⁸⁸ Re-DTPA-DG injection, the tumor inhibition rate was 29.2%. ¹⁸⁸ Re-DTPA-DG can inhibit tumor growth and play a therapeutic role.

Table 4 Comparison of tumor volume in nude mice with tumors in each group after treatment for 3 weeks (x±s) group Tumor volume (mm ³ ) saline group 1162.7±73.08 ReO ₄ ^-group 1153.0±99.42 ¹⁸⁸ Re-DTPA-DG group 823.6±50.58

5. ¹⁸⁸ Re-DTPA-DG induced apoptosis in lung cancer A549 cell line:

The lung cancer A549 cell line in the logarithmic growth phase was inoculated in a 24-well culture plate, placed in a 5% CO ₂ incubator at 37° C. for 24 hours, and then the corresponding solution was added. Divided into ReO ₄ ^-group and ¹⁸⁸ Re-DTPA-DG group, add 1mL ReO ₄ ^- 37, 55.5, 74kBq/mL, 1mL ¹⁸⁸ Re-DTPA-DG 37, 55.5, 74kBq/mL respectively. After adding the drug, culture in a 37°C 5% CO ₂ incubator for 18 hours, collect the cells by blowing mechanically, and follow the operation steps of the flow cytometry kit, count 10 ⁵ cells each time, and detect the percentage of apoptosis and the peak of apoptosis.

After cell counting, the tumor cell apoptosis percentage was detected (Table 5). There was a significant difference between the different specific activity ReO ₄ ^-groups and the ¹⁸⁸ Re-DTPA-DG group (P<0.01), and the ReO ₄ ^-group and ^{the 188} Re- There were significant differences among different specific activities in DTPA-DG group (P<0.01). ¹⁸⁸ Re-DTPA-DG can increase the apoptosis of tumor cells and can effectively treat tumors.

Table 5 Apoptotic percentage of lung cancer A549 cells in different groups (x±s) group Apoptotic percentage (%) 37kBq/mL 55.5kBq/mL 74kBq/mL ¹⁸⁸ Re-DTPA-DG group 22.4±4.51 22.9±2.18 40.0±5.6 ReO ₄ ^-group 4.3±1.29 5.2±1.65 9.3±1.24

¹⁸⁸ Re-DTPA-DG can be selectively taken up by tumor tissue after injection, while the uptake by non-tumor tissue is low, which can improve the targeting of Re treatment of tumors, with good curative effect, less side effects, and significantly improved prognosis. Oncology provides new methods of internal radiation therapy. ¹⁸⁸ Re-DTPA-DG is simple to prepare, easy to use, low in price, and can be supplied at any time. It will be an ideal new drug for internal radiation therapy of tumor metabolism and has broad clinical application prospects.

Therefore, the radioactive rhenium-labeled DTPA-DG complex of the present invention has a simple preparation method and low cost, can be used for tumor imaging and treatment, and is expected to become a new tumor therapeutic agent and imaging agent with good prospects.

Example 2:

In the acid chloride bisanhydride (0.010mol) obtained in the manner of Example 1, add 40mlDMSO, 8ml pyridine and 7.55g (0.035mol) 2-amino-D-glucose hydrochloride, and the mixture was stirred at 20°C-140°C for 40 hours , a transparent brown solution was obtained. This solution is separated and purified by dialysis membrane or Sephadex (G15), etc. to obtain the product (DTPA-3DG) in which R ₁ , R ₂ and R ₃ are all D-glucosamine-2 in the structure of formula (II).

Refer to the method for preparing ¹⁸⁸ Re-DTPA-3DG complexes with the resulting DTPA-3DG product

Example 1.

Example 3:

In the acid chloride bisanhydride (0.010mol) obtained in the manner of Example 1, 30mlDMSO, 6ml pyridine and 2.35g (0.012mol) 2-mercapto-D-glucose were added, and the mixture was stirred at -10°C-120°C for 40 hours to obtain brown solution. This solution is separated and purified by dialysis membrane or Sephadex (G10), etc., to obtain the structure product (DTPA-2DG) of formula (II) in which R ₁ and R ₂ are 2-mercapto-D-glucose, and R ₃ is H.

Refer to the method for preparing ¹⁸⁸ Re-DTPA-2DG complexes with the resulting DTPA-2DG product

Example 1.

Example 4:

Obtain DTPA-DG in the manner of Example 1, and use the obtained DTPA-DG product to prepare ¹⁸⁶ Re-DTPA-DG complex with reference to Example 1, except that SnCl ₂ 2H ₂ O is changed to 5 mg, radioactive perrhenate Change it to 100-500μL.

Example 5:

Obtain DTPA-3DG according to the method of Example 2, and use the obtained DTPA-3DG product to prepare ¹⁸⁶ Re-DTPA-3DG complex with reference to Example 1, except that SnCl ₂ 2H ₂ O is changed to 5 mg, radioactive perrhenate Change it to 100-500μL.

Embodiment 6:

Obtain DTPA-2DG according to the method of Example 3, and use the obtained DTPA-2DG product to prepare ¹⁸⁶ Re-DTPA-2DG complex with reference to Example 1, except that SnCl ₂ 2H ₂ O is changed to 5 mg, radioactive perrhenate Change it to 100-500μL.

Claims (4)

1, a kind of title complex that is used for tumor imaging and treatment, chemical structure of general formula is shown in (I):

A is 186 or 188 in the formula (I), R ₁Be sugar or osamine, R ₂And R ₃Difference H and/or sugar or osamine.

2, a kind of method for preparing title complex described in the claim 1 is characterized in that comprising the steps:

In closed reaction vessel, add reaction part, weak complexing agent, radioactive rhenium and reductive agent; Conditioned reaction system pH value is between 2-5.5; Heating in water bath makes temperature of reaction at 20-100 ℃, placing response 30-180 minute; Used reaction part is any one among DTPA-DG, DTPA-2DG, the DTPA-3DG, and used weak complexing agent is a Sunmorl N 60S, and used reductive agent is the saturated SnC of rare gas element ₁₂2H ₂O solution; Be reflected under the protection of inert gas and carry out.

3, method according to claim 2 is characterized in that reductive agent SnC ₁₂2H ₂The consumption of O solution is 0.5-10mg/ml, and rare gas element is a nitrogen.

4, method according to claim 2 is characterized in that described rare gas element is any one in helium, neon, argon, the nitrogen.

Images