KR101829913B1 - Radioisotope-labelled benzothiazole derivatives and a radiopharmaceutical product comprising the same - Google Patents

Abstract

The present invention relates to a compound that binds a DOTA ligand to a benzothiazole and binds a PET diagnostic radioactive isotope or a cancer therapeutic radioactive isotope via the ligand, and a PET contrast agent or a radiopharmaceutical for cancer treatment do.

Description

TECHNICAL FIELD The present invention relates to radioisotope-labeled benzothiazole derivatives and a radiopharmaceutical product comprising the same, and a radiolabeled benzothiazole derivative and a derivative thereof,

TECHNICAL FIELD The present invention relates to a radioactive metal-labeled benzothiazole derivative, a process for producing the same, and a radiopharmaceutical including the derivative, and more particularly, to a novel radioactive metal-labeled benzothiazide which can be used as a PET contrast agent or a cancer treatment agent. An azole derivative, a preparation method thereof, and a radiopharmaceutical including the derivative.

Positron Emission Tomography (PET) is a device that images the distribution of radiopharmaceuticals injected into a living body. It images the biological changes of the human body caused by diseases and provides accurate information for early diagnosis of diseases and methods of treatment of diseases. . Therefore, the importance of nuclear medicine imaging devices is increasing.

⁶⁸ Ga, ⁶⁴ Cu, and ⁸⁶ Y are typical metallic radioactive isotopes used for PET (Positron Emission Tomography) imaging in nuclear medicine. There are also ¹⁷⁷ Lu, ⁹⁰ Y, ¹⁸⁸ Re, and ⁶⁷ Cu radioisotopes that have been used to treat tumors with radiation in the field of nuclear medicine.

Among the radioisotopes capable of positron emission, relatively recently developed metallic radioisotope [ ⁶⁴ Cu] copper (positron emissivity, about 34%) in the nuclear medicine field has been used as a beam form of accelerated cyclotron-enriched Ni-64 (P, n) Cu-64 nuclear reaction, and Cu-64 with a high radiochemical purity is required for clinical use. The high purity Cu-64 can be obtained by separating Cu, Ni and other metal impurities through the column using electrostatic properties. The Cu-64 thus purified is present as a divalent cation on a diluted aqueous hydrochloric acid solution, and is characterized in that it can easily coordinate with a chelate having a negative charge. In addition, since Cu-64 has a relatively long half-life of 12.7 hours due to the physical properties of Cu-64, there is an advantage that images can be obtained after a long time after the injection, which could not be obtained with the conventional half-life radiolabeled radioactive pharmaceutical within 2 hours.

In radiotherapy using ionizing radiation, it is very important to evaluate the accurate dose of radiation absorbed into the tumor and normal tissues of the radiopharmaceutical administered to the patient. Because radiopharmaceuticals remain in the body when injected into a patient's body and release relatively low doses of radiation over a long period of time. Therefore, in order to determine the therapeutic dose using the image-based dose evaluation, a small amount of radiopharmaceutical is actually injected before the treatment, the radiation dose is calculated from the image test, and an appropriate amount of radiation And to determine the therapeutic injection dose to be administered.

PET is currently the most accurate method of assessing radioactivity accumulation in tissue. PET is sensitive to extinction photons and has better sensitivity than SPECT, and quantification method is well established. It is ideal to use isotopes such as isotopes to be used in PET imaging for dose assessment. For example, in the case of treatment with ⁶⁷ Cu, a PET image using ⁶⁴ Cu can be used (non-patent reference 1), and in case of ⁹⁰ Y treatment, a dose evaluation using ⁸⁶ Y can be performed. However, due to the limitations of available radioisotopes depending on the hospital environment and various circumstances, it is practically difficult to use the same radioactive isotope and the same ligand structure in imaging and treatment for dose assessment. Therefore, a dose evaluation for ⁹⁰ Y is performed using ¹¹¹ In (Non-Patent Document 2) or a dose evaluation for ¹⁷⁷ Lu is performed using ⁶⁸ Ga (Non-Patent Document 3).

Accurate dose assessment in radioactive isotope therapy is necessary to achieve effective doses at the treatment site and to reduce radiation exposure to the normal site.

PET is a valuable tool for understanding biological phenomena or for accurately diagnosing various tumors, i) high selectivity of radiopharmaceuticals for the tumor, ii) fast intake and equilibration time after injection, iii) imaging (See Non-Patent Document 4).

On the other hand, benzothiazole has a selective anticancer effect against cancer cells, and various types of derivatives have been applied as anticancer substances in clinical or preclinical studies (Non-Patent Documents 5 and 6).

1. Shen, S. et al. J. Nucl. Med., 1996, 37 (1), 146-150. 2. L ㆆ ovqvist, A. et al. J. Nucl. Med., 2001, 42 (8), 1281-1287. 3. Aurora Sainz-Esteban et al. Eur J Nucl Med Mol Imaging 2012 39, 501-511. 4. Carolyn J. Anderson, Riccardo Ferdani, Copper-64 Radiopharmaceuticals for PET Imaging of Cancer: Advances in Preclinical and Clinical Resaerch. Cancer Biotherapy and Radiopharmaceuticals. 2009, 24, 379-393 5. Ian Hutchinson et al. J. Med. Chem., 2002, 45, 744-747. 6. T. D. Bradshaw et al. Current Medicinal Chemistry, 2004, 11 1241-1253.

One aspect of the present invention is to provide a radioisotope-labeled benzothiazole derivative which can be used as an effective PET contrast agent or cancer treatment agent because of its high binding affinity for tumors.

Another aspect of the present invention is to provide a PET contrast agent comprising a diagnostic radioisotope-labeled benzothiazole derivative.

Another aspect of the present invention is to provide a radiopharmaceutical for treating cancer comprising a therapeutic radioisotope-labeled benzothiazole derivative.

Another aspect of the present invention is to provide a radiopharmaceutical for cancer treatment which is capable of evaluating a radiation dose using a PET image.

One aspect of the present invention provides compounds of formula 1:

[Chemical Formula 1]

In the above formula (1), M is ⁶⁸ Ga, ⁶⁴ Cu, ¹⁷⁷ Lu, or ⁹⁰ Y.

Another aspect of the present invention provides a PET contrast agent comprising a compound of formula (I): < EMI ID =

[Formula 1a]

In Formula 1a, M is ⁶⁸ Ga or ⁶⁴ Cu.

Another aspect of the present invention provides a radiopharmaceutical for treating cancer comprising a compound of the following formula:

[Chemical Formula 1b]

In the above formula (1b), M is ¹⁷⁷ Lu or ⁹⁰ Y.

The compound of formula (I) according to one aspect of the present invention is excellent in selectivity for cancer and is stable in serum, and can be used as safe and effective PET contrast agent and radiopharmaceutical for cancer treatment.

In addition, the radiopharmaceutical for cancer treatment, which includes both the compound of formula (Ia) containing the radioactive isotope for PET imaging and the compound of formula (Ib) containing the compound for radioactive isotypes for cancer treatment, Can be monitored by the image, and the cancer treatment status can be checked, which is preferable. In addition, the compound of formula (Ia) and the compound of formula (Ib) may have a pair of radioisotopes that can exhibit substantially the same distribution, and it is preferable to determine whether the radiopharmaceutical for cancer treatment is distributed in a desired cancer site, Radiation therapy can be performed, and thus radiation therapy can be safely and effectively performed. In addition, the radiation dose in cancer tissue can be evaluated by PET imaging using formula (Ia), and the dose of the compound of formula (Ib) suitable for the living body to be administered can be determined based on the radiation dose.

Figure 1 is a graph showing the uptake rate of ⁶⁸ Ga-DOTA-BTA for various types of four tumor cells.
Figure 2 is a graph showing the uptake rates of ¹⁷⁷ Lu-DOTA-BTA and ¹⁷⁷ Lu-DOTA to HeLa cells.
FIG. 3 is a graph showing the time-based comparison of the tissue radioactivity ratio (% ID / g) determined by injecting ⁶⁸ Ga-DOTA-BTA into a mouse having a HeLa cell tumor and measuring various organs.
FIG. 4 is a graph showing the time-based comparison of the tissue radioactivity ratio (% ID / g) measured in various organs to injected ⁶⁴ Cu-DOTA-BTA into HeLa cell tumor mice.
FIG. 5 is a graph showing the time-based comparison of the tissue radioactivity ratio (% ID / g) measured in various organs to injected ¹⁷⁷ Lu-DOTA-BTA into mice having HeLa cell tumors.
FIG. 6 is a PET image obtained after the injection of ⁶⁸ Ga-DOTA-BTA into a mouse having a HeLa cell tumor.
Fig. 7 is a PET image obtained by injecting ⁶⁴ Cu-DOTA-BTA into a mouse with HeLa cell tumor and lapse of time.
8 is a graph showing the results of comparing the survival rates of ¹⁷⁷ Lu-DOTA-BTA and ¹⁷⁷ Lu-DOTA to HeLa cells at various doses.
[Description of Drawings]
Radiopharmaceutical uptake: Radioactive drug uptake rate
Survival: Survival rate

Hereinafter, the present invention will be described in more detail.

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. In addition, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications referred to in this specification are incorporated herein by reference in their entirety.

The present inventors have studied to develop an effective and safe radiopharmaceutical for cancer diagnosis or cancer treatment and have developed a PET contrast agent compound in which a DOTA ligand is bound to a benzothiazole and a PET diagnostic radioisotope is bound to the ligand. Compounds have excellent selectivity for tumor and excellent stability in serum, and thus can be used as an effective and safe PET contrast agent. Further, a cancer therapeutic compound was prepared by binding a DOTA ligand of the above compound to a radioactive isotope for cancer treatment instead of the PET diagnostic radioisotope, and the compound was excellent in selectivity for tumor and excellent in serum stability, (Examples 5, 6, and 7). ≪ / RTI >

Accordingly, in one aspect, the present invention provides a compound of formula 1:

[Chemical Formula 1]

In the above formula (1), M is ⁶⁸ Ga, ⁶⁴ Cu, ¹⁷⁷ Lu, or ⁹⁰ Y.

The compounds of formula 1 may be prepared, for example, as described in Examples 2 to 4 below, and the preparation method thereof may be appropriately modified by those skilled in the art of organic chemistry using known organic chemistry knowledge .

And in the compound of Formula 1, may be used as a diagnostic PET contrast agents according to the kind of the radioisotope M, can also be used as therapeutic radiopharmaceuticals for for the treatment of cancer, specifically, M is a diagnostic radioisotope ⁶⁸ Ga Or ⁶⁴ Cu, and can be used as a diagnostic PET contrast agent, and when M is the therapeutic radioactive isotope ¹⁷⁷ Lu or ⁹⁰ Y, it can be used as a radiopharmaceutical for cancer treatment.

In another aspect, the present invention provides a PET contrast agent comprising a compound of formula (I): < EMI ID =

 [Formula 1a]

In Formula 1a, M is ⁶⁸ Ga or ⁶⁴ Cu.

Hereinafter, the PET contrast agent is also referred to as PET diagnostic radiopharmaceutical.

In one embodiment, the PET contrast agent is a PET contrast agent for cancer diagnosis.

The compounds of formula (Ia) are not only highly selective for tumor cells, but also have high stability for a long time in serum, and thus can be used as safe and effective contrast agents for diagnosis of cancer (see Examples 7-9, 11 and 12).

In one embodiment, the PET contrast agent is a PET contrast agent for cervical cancer diagnosis. As a result of the experiment, the compound of formula (Ia) was found to have a significantly higher uptake rate to Hela cells, especially cervical cancer cells, among various cancer cells (see Example 5).

The PET contrast agent may be administered to an adult in an amount of 0.1 to 30 mCi based on the compound of formula (I) as the active ingredient.

In another aspect, the present invention provides a radiopharmaceutical for treating cancer comprising a compound of the following formula (1b):

[Chemical Formula 1b]

In the above formula (1b), M is ¹⁷⁷ Lu or ⁹⁰ Y.

In one embodiment, the radiopharmaceutical for cancer treatment is a radiopharmaceutical for the treatment of cervical cancer.

The compound of formula (1b) has high selectivity for tumor cells and has high stability for a long time in serum, so that it can be used as safe and safe radiotherapeutic drug for cancer treatment (see Examples 7, 10, and 13).

The cancer therapeutic radiopharmaceutical may be administered to an adult in an amount of 20-50 mCi based on the compound of the formula (Ib) as the active ingredient.

In one embodiment, the radiopharmaceutical for cancer treatment is a radiopharmaceutical effective for the treatment of cervical cancer. As a result of the experiment, the compound of the formula (1b) was found to be significantly superior to Hela cells of the cervical cancer cells among various cancer cells (see Example 6).

The dose of the radiopharmaceutical for cancer treatment can be determined from the dose evaluation data using the PET image using the PET contrast agent containing the compound of formula (I).

Alternatively, the radiopharmaceutical for cancer treatment may further comprise a compound of formula (I) wherein M is ⁶⁸ Ga or ⁶⁴ Cu, and may be a radiopharmaceutical for cancer treatment capable of evaluating a dose using a PET image. By further including the compound of formula (Ia), it is possible to monitor the progress of the cancer treatment as well as the dose evaluation using the PET image.

The accurate dose of the radioisotope treatment can be evaluated by the dose evaluation using the PET image, and thus it is possible to attain an effective dose of radiation to the treatment site and to reduce the radiation exposure to the normal site.

The compound of formula (Ia) wherein M is ⁶⁸ Ga or ⁶⁴ Cu can be used as a diagnostic PET contrast agent for cancer as described above. Therefore, the radiotherapeutic drug for cancer treatment may be prepared by using PET image of formula (1a) And the dose of the therapeutic radiopharmaceutical, formula 1b, can be determined after a tissue dose assessment is performed. Thus, doses reaching normal tissues as well as tumors can be performed, which can help determine the dose that is effective in tumor treatment and minimizes radiation exposure to normal tissue.

In one embodiment, the radiopharmaceutical for cancer treatment capable of evaluating the dose comprises a compound of Formula 1b wherein M is ⁷⁷ Lu, and the compound of Formula 1a is a compound wherein M is ⁶⁸ Ga or ⁶⁴ Cu , And radiotermic medicines for cancer treatment that can be evaluated by PET imaging.

In another embodiment, the PET contrast agent comprising a compound wherein M is ⁶⁸ Ga or ⁶⁴ Cu as a compound of formula (Ia) is used in a dose assessment of a radiopharmaceutical for treating cancer comprising a compound wherein M is ⁷⁷ Lu as a compound of formula . ≪ / RTI >

These compounds of formula (Ia) and (Ib) may exhibit substantially similar bodily distributions (see examples 8, 9, and 10). Therefore, the radiopharmaceutical for cancer treatment can evaluate the radiation dose in cancer tissue using PET images, and the dose of the compound of formula (Ib) suitable for a living body to be administered based on the radiation dose can be determined.

The radiopharmaceutical for PET diagnosis and the radiopharmaceutical for cancer treatment according to the present invention may be formulated as an injectable preparation. When formulated into an injectable preparation, a non-toxic buffer solution appearing as blood may be contained as a diluent. For example, Buffer solution and the like. The PET diagnostic radiopharmaceuticals and radiopharmaceuticals for cancer treatment may contain other diluents or additives in addition to the buffer solution. Excipients and additives that may be added to such injections are well known to those skilled in the art and can be found, for example, in the following references: Dr. HP Fiedler "Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik und angrenzende Gebiete "[Encyclopedia of auxiliaries for pharmacy, cosmetics and related fields]).

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, these examples and experimental examples are provided to aid understanding of the present invention, and the scope of the present invention is not limited thereto in any sense.

Abbreviation definitions

DCM: Dichloromethane

DMF: N, N-dimethylformamide

ACN: Acetonitrile

DIEA: Diisopropylethylamine

DOTA: 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid

BTA: benzothiazole

PBS: Phosphate Buffer's Solution

TLC: Thin layer chromatography

Example 1: Preparation of DOTA-BTA

DOTA-BTA was prepared according to Reaction Scheme 1 below.

[Reaction Scheme 1]

A. Preparation of tert-butyl 2,2 ', 2'- (10- (2- (4- (benzo [d] thiazol-2- yl) phenylamino) -2-oxoethyl) tetraazacyclododecane-1,4,7-triyl) triacetate

^{_{DO 3 A (t Bu) 3}} (1.1 g, 2.2 mmol) and _{NaHCO 3 (0.5 g, 6.6 mmol} ) to a solution in DMF (10 mL) N - ( 4- (benzo [d] thizol-2-yl ) phenyl) -2-chloroacetamide (1.0 g, 3.3 mmol) was dissolved in DMF (10 mL). After 18 hours of reaction at room temperature, water (200 mL) was added and the reaction was further continued for 2 hours. The precipitate was filtered and washed with water. The precipitate was separated by silica column chromatography (silica, CH ₂ Cl ₂ / MeOH, 97: 3) to obtain a white precipitate.

Yield: 0.5 g (30%).

_{^{1 H NMR (CDCl 3):}} δ = 8.09-8.07 (d, 2H, phenyl), 8.02-8.00 (d, 1H, benzothiazole), 7.92-7.90 (d, 2H, phenyl), 7.86-7.85 (d, 1H _{, benzothiazole), 7.47-7.43 (t d} , 1H, benzothiazole), 7.36-7.32 (t d, 1H, benzothiazole), 3.05-2.85 (m, 8H, -C H 2 CO 2, -C H 2 = CO) , 2.61-2.02 ( m , 16H, C H ₂ in the cyclen ring).

Anal. Calcd for C ₄₁ H ₆₄ N ₆ O ₇ S 4 HCl: C, 53.13 H, 6.96 N, 9.07; S, 3.46. Found: C, 52.93 H, 6.62 N, 8.97; S, 3.17.

HR-FABMS (m / z) : calcd for C 41 H 60 N 6 O 7 SNa: 803.4142 ([MNa] +). Found: 803.4142 ([MNa] < ⁺ >).

B. Preparation of BTA-DOTA

tert-butyl 2,2 ', 2'- (10- (2- (4- (benzo [d] thiazol-2-yl) phenylamino) -2-oxoethyl) -1,4,7,10-tetraazacyclododecane- 1,4,7-triyl) triacetate (1.0 g, 1.3 mmol) was dissolved in a mixture of TFA (Trifluoroacetic Acid) / CH ₂ Cl ₂ (1: 1, 20 mL) and stirred for 24 hours. Dissolved in MeOH (10 mL), and then precipitated in diethyl ether (100 mL).

Yield: 0.6 g (68%)

_{^{1 H NMR (Methanol-d 4}} ): δ = 8.00-7.95 (m, 4H, phenyl), 7.79-7.77 (m, 2H, benzothiazole), 7.53-7.50 (t, 1H, bnezothiazole), 7.43-7.40 (t , 1H, bnezothiazole), 3.86-3.61 ( m , 8H, -C H ₂ CO ₂ , -C H ₂ = CO), 3.50-3.14 ( m , 16H, C H ₂ in the cyclen ring).

Anal. Calcd for C ₂₉ H ₄₆ N ₆ O ₁₂ S 5H ₂ O: C, 49.56 H, 6.60 N, 11.96; S, 4.56. Found: C, 49.57 H, 6.20 N, 12.33; S, 4.45.

HR-FABMS ( m / z ): calcd for C ₂₉ H ₃₆ N ₆ O ₇ SNa: 635.2264 ([MNa] ⁺ ). Found: 635.2264 ([MNa] < ⁺ >). HPLC (analytical: 2.1 x 250 mm, flow rate: 1.4 min / L, retention time: 24.12 min).

Example 2: ⁶⁸ Fabrication of Ga-DOTA-BTA

⁶⁸ the Ge / ⁶⁸ Ga a ⁶⁸ Ga eluted from a generator was obtained as a clear solution of the binary dissolved in dilute hydrochloric acid solution was colorless, after obtained as appropriate to the radioactivity necessary for the experiment (1 or more mCi) ⁶⁸ Ga solution in an appropriate glass vessel Followed by drying at 100 DEG C while blowing nitrogen. The dried ⁶⁸ Ga is attached to the glass container in the form of a transparent film. Subsequently, 0.5 mg of BTA-DOTA was dissolved in 0.5 mL of 1M sodium acetate buffer solution (pH 5-6) to obtain about 100 μg / 0.01 mL. The mixture was mixed with the ⁶⁸ Ga solution and the labeling reaction was allowed to proceed at 100 ° C. for 5 minutes. The final product, the BTA-DOTA- to adsorb the ⁶⁸ Ga thin layer chromatography confirmed the fully deployed after the radiochemical yield and purity as eluent (0.1M citrate buffer).

Example 3: ⁶⁴ Preparation of Cu-DOTA-BTA

⁶⁴ Cu produced from the accelerator is obtained as a colorless transparent solution dissolved in a diluted hydrochloric acid solution. After appropriately obtaining the amount of radioactivity required for the experiment (0.1 mCi or more), ⁶⁴ Cu solution is put in a suitable glass container and nitrogen is blown And dried at 100 ° C. The dried ⁶⁴ Cu was attached to the glass vessel in the form of a transparent membrane, and 0.2-0.3 mL of 1 M sodium acetate buffer solution (pH 5-6) was added and dissolved. Subsequently, BTA-DOTA was dissolved in ethanol to obtain 100 μg / 0.01 mL of the solution. The resultant mixture was mixed with the ⁶⁴ Cu solution and the labeling reaction was allowed to proceed at 50 ° C. for 30 minutes. The final product, BTA-DOTA- ⁶⁴ Cu, was adsorbed on thin layer chromatography and developed sufficiently with 0.1 M citrate buffer solution to confirm the radiochemical yield and purity.

Example 4: ¹⁷⁷ Manufacture of Lu-DOTA-BTA

The ¹⁷⁷ Lu produced from the reactor is obtained as a colorless transparent solution dissolved in a dilute hydrochloric acid solution. The ¹⁷⁷ Lu solution is appropriately obtained according to the amount of radioactivity required for the experiment (0.1 mCi or more), and ¹⁷⁷ Lu solution is placed in a suitable glass container, And dried at 100 ° C. The dried ¹⁷⁷ Lu is attached to the glass container in the form of a transparent film. Subsequently, 0.5 mg of BTA-DOTA was dissolved in 0.5 mL of 1 M sodium acetate buffer solution (pH 5-6) to obtain 100 μg / 0.01 mL of the solution. The solution was mixed with the ¹⁷⁷ Lu solution and the labeling reaction was allowed to proceed at 100 ° C. for 5 minutes. The final product, BTA-DOTA- ¹⁷⁷ Lu, was adsorbed on the thin layer chromatography and sufficiently developed with a developing solution (1 M sodium hydroxide: ethanol: distilled water mixed 1: 5: 10) Respectively.

Example 5: ⁶⁸ Determination of tumor cell selectivity of Ga-DOTA-BTA

The selectivity of ⁶⁸ Ga-DOTA-BTA prepared in Example 2 for tumor cells was examined. Four types of tumor cells were used: human cervix adenocarcinoma cell line (HeLa), human embryonic kidney cell line (293), human breast cancer cell line (MDA-MA-231) and human glioblastoma cell line (U87MG).

Each of the four tumor cells was cultured in a dish, and ⁶⁸ Ga-DOTA-BTA was added thereto. After washing for 24 hours, the amount of ⁶⁸ Ga-DOTA-BTA that the cells ingested was measured by a gamma counter. The results are shown in Fig.

FIG. 1 is a graph showing the uptake rate of ⁶⁸ Ga-DOTA-BTA of the four tumor cells. According to the results shown in Fig. 1, ⁶⁸ Ga-DOTA-BTA was found to be significantly larger than other tumor cells, especially in HeLa cells, in cancer cells.

Example 6: ¹⁷⁷ Identification of tumor cell selectivity of Lu-DOTA-BTA

The uptake rates of the ¹⁷⁷ Lu-DOTA-BTA and ¹⁷⁷ Lu-DOTA prepared in Example 4 on the HeLa cell line were compared in the same manner as in Example 5 above. The purpose of this experiment is to compare the selectivity for tumor cells with or without BTA. The results are shown in Fig.

Figure 2 is a graph showing the uptake rates of ¹⁷⁷ Lu-DOTA-BTA and ¹⁷⁷ Lu-DOTA to HeLa cells. 2, the binding capacity of ¹⁷⁷ Lu-DOTA-BTA to HeLa cells was significantly greater than that of ¹⁷⁷ Lu-DOTA.

Example 7: ¹⁷⁷ With Lu-DOTA-BTA ⁶⁴ Confirmation of in-vitro serum stability of Cu-DOTA-BTA

The stability of ¹⁷⁷ Lu-DOTA-BTA and ⁶⁴ Cu-DOTA-BTA prepared in Examples 3 and 4 in human serum and mouse serum was measured. ¹⁷⁷ Lu-DOTA-BTA and ⁶⁴ Cu-DOTA-BTA (0.1 mCi, 10 mL) were separately mixed with human serum, mouse serum (0.5 mL) . TLC was performed on the time to be measured. The stability of ¹⁷⁷ Lu-DOTA-BTA and ⁶⁴ Cu-DOTA-BTA in serum was shown in Table 1 below.

[Table 1]

According to the results in Table 1 above, both ¹⁷⁷ Lu-DOTA-BTA and ⁶⁴ Cu-DOTA-BTA were stable for 36 hours in human and mouse serum and PBS.

Example 8: ⁶⁸ Identification of Ga-DOTA-BTA tissue distribution

The distribution of mouse tissues of ⁶⁸ Ga-DOTA-BTA prepared in Example 2 was confirmed. HeLa cells were subcutaneously injected into the thighs of nude mice (BW 20 g) to form tumors. The mice were injected with 0.01 mCi of ⁶⁸ Ga-DOTA-BTA through the tail vein of the tumor-bearing mouse (n = 4), and then the tumor was treated with 1 hour, 2 hours, , Kidney, spleen, bone, muscle, intestine) were measured and the radioactivity of the tissue was measured with a gamma counter.

FIG. 3 is a graph showing time-based comparison of the tissue radioactivity ratio (% ID / g) measured in various organs after injecting ⁶⁸ Ga-DOTA-BTA according to an embodiment of the present invention.

3 shows that the radioactivity present in the tumor is maintained at 3% or more at 1 hour after injection and at 2% or more at 4 hours after injection.

Example 9: ⁶⁴ Confirmation of Cu-DOTA-BTA tissue distribution

The distribution of mouse tissues of ⁶⁴ Cu-DOTA-BTA prepared in Example 3 was confirmed. HeLa cells were subcutaneously injected into the thighs of nude mice (BW 20 g) to form tumors. The mice were injected with 0.01 mCi of ⁶⁴ Cu-DOTA-BTA via the tail vein of the tumor-bearing mouse (n = 4), and the organs (blood, tumor, brain, heart, lung, liver , Kidney, spleen, bone, muscle, intestine) were measured and the radioactivity of the tissue was measured with a gamma counter.

FIG. 4 is a graph showing the time-based comparison of the tissue radioactivity ratio (% ID / g) measured in various organs after injecting ⁶⁴ Cu-DOTA-BTA according to an embodiment of the present invention.

4, it can be seen that the radioactivity present in the tumor is maintained at 3% or more at 1 hour after injection and at 2% or more at 36 hours after injection.

Example 10: ¹⁷⁷ Identification of Lu-DOTA-BTA tissue distribution

The distribution of mouse tissues of ¹⁷⁷ Lu-DOTA-BTA prepared in Example 4 was confirmed. HeLa cells were subcutaneously injected into the thighs of nude mice (BW 20 g) to form tumors. After injection of 0.01 mCi of ¹⁷⁷ Lu-DOTA-BTA through the tail vein of the tumor-bearing mouse (n = 4), the organs (blood, tumor, brain, heart, lung, liver , Kidney, spleen, bone, muscle, intestine) were extracted and the radioactivity of the tissue was measured with a gamma counter.

FIG. 5 is a graph showing time-based comparison of the tissue radioactivity ratio (% ID / g) measured in various organs after injecting ¹⁷⁷ Lu-DOTA-BTA prepared according to an embodiment of the present invention.

5, it can be seen that the radioactivity present in the tumor is maintained at 2.7% or more at 1 hour after injection and at 1.5% or more at 4 hours after injection.

Example 11: ⁶⁸ PET image acquisition using Ga-DOTA-BTA

Tumor model mice were treated with ⁶⁸ Ga-DOTA-BTA prepared in Example 2 to obtain PET images. PET images were taken with multiple PET / SPECT / CT systems (INVEON, Simens Medical Solutions).

The tumor model was obtained by transplanting human cervix adenocarcinoma cell line (HeLa) into the thigh of nude mice and imaging when the size of tumor was ~ 0.5 g. ⁶⁸ Ga-DOTA-BTA 0.5 mCi, and PET images were obtained with the passage of time. The results are shown in Fig.

According to FIG. 6, the PET image obtained after administration of ⁶⁸ Ga-DOTA-BTA showed an increase in signal at the tumor site (the color became bright) and continued signaling for up to 4 hours after administration.

Example 12: ⁶⁴ PET image acquisition using Cu-DOTA-BTA

Tumor model mice were treated with ⁶⁴ Cu-DOTA-BTA prepared in Example 3 to obtain PET images. PET images were taken with multiple PET / SPECT / CT systems (INVEON, Simens Medical Solutions).

The tumor model was obtained by transplanting human cervix adenocarcinoma cell line (HeLa) into the thigh of nude mice and imaging when the size of tumor was ~ 0.5 g. ⁶⁴ Cu-DOTA-BTA 0.2 mCi, and the PET image was acquired with the lapse of time. The results are shown in Fig.

According to Fig. 7, the PET image obtained after administration of ⁶⁴ Cu-DOTA-BTA showed an increase in signal at the tumor site (the color became bright) and the signal persisted for up to 48 hours after administration.

Example 13: ¹⁷⁷ Investigation of in vitro tumor treatment effect of Lu-DOTA-BTA

The cancer cell inhibitory effect of ¹⁷⁷ Lu-DOTA-BTA prepared in Example 4 was tested. HeLa cells were treated with 1.85 MBq, 3.7 MBq, and 9.25 MBq of ¹⁷⁷ Lu-DOTA-BTA and ¹⁷⁷ Lu-DOTA, respectively, and cell viability was observed after 24 h. This experiment is also intended to compare the therapeutic effect of tumor cells on the presence or absence of BTA. The results are shown in Fig.

According to Fig. 8, the group treated with ¹⁷⁷ Lu-DOTA-BTA or ¹⁷⁷ Lu-DOTA showed significantly lower cell survival rate than the control group. In addition, the higher the dose of ¹⁷⁷ Lu-DOTA-BTA or ¹⁷⁷ Lu-DOTA, the lower the cell survival rate. In addition, the group containing ¹⁷⁷ Lu-DOTA-BTA showed significantly lower cell survival rate than the group containing ¹⁷⁷ Lu-DOTA. These results indicate that ¹⁷⁷ Lu-DOTA-BTA is significantly superior to the cancer cell suppression effect due to selective action on tumor cells.

Claims (9)

A pharmaceutical composition for diagnosis or treatment of cervical cancer comprising the compound of formula
[Chemical Formula 1]

In the above formula (1), M is ⁶⁸ Ga, ⁶⁴ Cu, ¹⁷⁷ Lu, or ⁹⁰ Y. A PET contrast agent for cervical cancer diagnosis comprising a compound of formula
[Formula 1a]

In Formula 1a, M is ⁶⁸ Ga or ⁶⁴ Cu. delete delete Radiopharmaceuticals for the treatment of cervical cancer comprising the compound of formula (Ib):
[Chemical Formula 1b]

In the above formula (1b), M is ¹⁷⁷ Lu or ⁹⁰ Y. delete The radiopharmaceutical for the treatment of cervical cancer according to claim 5, which further comprises a compound of formula (I):
[Formula 1a]

In Formula 1a, M is ⁶⁸ Ga or ⁶⁴ Cu. The radiopharmaceutical for the treatment of cervical cancer according to claim 7, wherein M in the formula (1b) is ¹⁷⁷ Lu. 8. The radiopharmaceutical for the treatment of cervical cancer according to claim 7, wherein M in the formula (1b) is ¹⁷⁷ Lu and M in the formula (1a) is ⁶⁴ Cu.

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