KR100806368B1 - Radio-labeled pyrazineamide composite and its kit as radiophamarceuticals for imaging tuberculosis - Google Patents

Abstract

The present invention relates to a tuberculosis nuclear medicine imaging contrast agent and a kit used to determine whether the tuberculosis bacillus is active by injecting an anti-tuberculosis antibiotic pyrazineamide combined with a radionuclide into an animal or human body. Relates to the preparation of tuberculosis nuclear medicine imaging contrast kits and kits prepared by labeling pyrazineamide with Tc-99m, Indium-111, Iodine-131 and / or Iodine-123.

Tuberculosis, Pulmonary Tuberculosis, Pyrazinamide, Pulmonary Tuberculosis Imaging Agent, Tc-99m, Indium-111, Iodine-131, Iodine-123

Description

Radio-labeled pyrazineamide composite and its kit as radiophamarceuticals for imaging tuberculosis

1 is a diagram showing the binding pyrazineamide (PZA) -I purity,

Figure 2 is a photograph showing the degree of binding pyrazineamide (PZA) -I bioabsorption,

3 and 4 are images taken with the tuberculosis bacteria infection nuclear medicine imaging kit according to the present invention and the corresponding x-ray picture.

The present invention relates to a tuberculosis nuclear medicine imaging contrast agent and a kit used to determine whether the tuberculosis bacillus is active by injecting an antituberculosis antibiotic pyrazineamide combined with a radionuclide into an animal or human body.

Existing assays are not easy to diagnose active pulmonary tuberculosis (Cegielski JP, Chin DP, Espinal MA, Frieden TR, Rodriquez Cruz R, Talbot EA, Weil DE, Zaleskis R, Raviglione MC) The global tuberculosis situation.Progress and problems in the 20th century, prospects for the 21st century.Infect Dis Clin North Am. 2002 Mar; 16 (1): 1-58 Cegielski, 2002). Normally, the existing inspection method is 1) to perform lung radiographing regularly to determine the progression of the disease through shadow reading in the photograph, but this diagnosis method has a problem of lack of objectivity and reproducibility. A method of determining a test by culturing a sample such as a sputum, but this has a disadvantage in that the incubation time is slow and the promptness is poor in determining treatment. In particular, in the case of inactive pulmonary tuberculosis, the administration of anti-tuberculosis drug is not necessary and should not be administered, but it is pointed out that the problem of anti-tuberculosis drug use is caused by the empirical use of anti-tuberculosis drug without the definite objective diagnosis method. In addition, insufficient time to use anti-tuberculosis drugs may result in a relapse of the disease. Therefore, it is important to have an accurate and objective diagnosis and to use antituberculosis drugs for a sufficient and necessary time.

According to Korean Patent Registration Publication No. 10-0242177 (imaging of infection, Applicant: Vintage International Limited), a means for early identification of infection using radionuclide-labeled antibiotics that specifically target bacterial infection sites is provided. Providing. According to this, metal radionuclides, 4-quinoline antibiotic compounds and pharmaceutically acceptable salts thereof are provided for use in gamma-ray scintography diagnostic imaging of patients, specifically ciprofloxacin, oploxacin and Norfloxacin is mentioned, and these quinoline antibiotics act as antibiotics by inhibiting the DNA gyrase action required for bacteria to actively divide (Hooper DC. Quinolone mode of action.Drugs. 1995; 49 Suppl 2: 10- 5) Therefore, antibiotics combined with radionuclides penetrate into living bacterial cells and emit gamma rays, allowing for nuclear medical imaging. Ciprofloxacin and the like according to the Korean Patent Registration Publication have been reported in clinical research on orthopedic infectious diseases (for example, osteomyelitis and purulent arthritis, etc.) (Malamitsi J, Christacopoulos J, Dounis E, et al. 99m Tc-ciprofloxacin ( infecton) as a means to detect bone and joint infection.Eur J Nucl Med. 1997; 24 (8): 876). However, in case of confining the infected area to pulmonary tuberculosis, unlike the third-line drug, ciprofloxacin, it belongs to the first-line drug and has excellent effect as a treatment for pulmonary tuberculosis (PZA, There is a need to use pyrazinamide).

Certain antibiotics are obviously not intended for use in nuclide imaging. Antibiotics are radioactive because they are produced for the purpose of achieving therapeutic use in most tissues of the body, rather than for the purpose of inducing increased resistance and sensitization in the body, and for the antibiotics to be absorbed purely at the site of infection. It is not clear if it can be converted into a diagnostic substance complexed with a nuclide.

The present invention relates to a tuberculosis nuclear medicine imaging agent and a kit used to determine whether a tuberculosis bacillus is infected by injecting the antibiotic pyrazineamide combined with a radionuclide into an animal or human body.

The present invention, in order to achieve the above object, a reducing step of mixing the antibiotic pyrazine amide and a reducing agent to produce a first mixture, a mixing step of mixing a radionuclide to the first mixture to produce a second mixture, The present invention provides a tuberculosis nuclear medicine imaging medium and its kit prepared by a binding step of binding pyrazineamide and radionuclide included in the second mixture. Mycobacterium tuberculosis can be infected not only the lung but also each part of the human body, such as bone, but for the purpose of illustrating the present invention to limit the infection site to the case of the lungs for specific examples of the present invention, the idea of the present invention is not limited to pulmonary tuberculosis. It is obvious that the present invention can be applied to the determination of the active and tuberculosis of Mycobacterium tuberculosis in each part of the human body, and it is apparent to those skilled in the art.

Here, pyrazine amide is a known primary anti-tuberculosis antibiotic and has a structure of the following general formula (1).

Although the following compounds may be mentioned as primary pulmonary tuberculosis antibiotics, it is determined that pyrazineamide, a substance of the present invention, may form a stable complex with radionuclides.

Isoniazid

Glyconiazide

Ethambutol

The pyrazineamide precursor according to the present invention can be synthesized through the following procedure.

The present inventors attached the iodine as described above, and shows the data measured the purity in Figure 1, the results of the basic animal experiment to see the biodistribution of the iodine-coupled compound is shown in FIG. That is, as shown in FIG. 2 (90 minutes after the injection), the synthesized compound was metabolized with ingestion in the liver and spleen. The animal was confirmed to have no symptoms showing toxicity.

In the reduction step, the first step according to the invention, the applicable reducing agent may be formamidine sulfinic acid or formamidine sulfonic acid. In particular, the mass ratio of the pyrazine amide and the reducing agent in the reduction step is preferably 2: 1. In the second step of the mixing step according to the present invention, the radionuclide may be any one of Tc-99m, Indium-111, Iodine-131, and Iodine-123, and the content of the isotope per mL of the first mixture is 30. It may also be characterized by a 50 mCi. In addition, the heating time in the third step of the bonding step is characterized in that the microwave oven 800W ~ 1,000W 12 seconds ~ 15 seconds.

In another aspect, the present invention relates to a pulmonary tuberculosis nuclear medicine imaging agent kit, which is combined with a radionuclide and used as a pulmonary tuberculosis infection nuclear medicine imaging agent, wherein the kit is pyrazineamide and toxic reduction thereof Reducing agents and radionuclides which are mixed to make.

The pulmonary tuberculosis infection nuclear medicine imaging agent kit herein refers to a pyrazineamide compound or a pharmaceutically acceptable salt thereof to form radionuclides and complexes selected from the group consisting of Tc-99m, Indium-111, Iodine-131 and Iodine-123. It is defined as a kit for manufacturing pulmonary tuberculosis nuclear medicine imaging contrast agent, included with the reducing agent so that it is preserved or transported in a vial.

Hereinafter, the present invention will be described in detail, but is not limited thereto.

Example 1 Preparation of Pneumococcal Tuberculosis Infection Nuclear Medicine Imaging Kit

1. Reduction step

First, 2 mg of pyrazineamide is administered to a 10 mL vial. Meanwhile, formamidine sulfinic acid is added to a 10 mL vial and diluted with distilled water to a concentration of 1 mg / 100 μL. Then dissolve in Vortex-Gene2 until completely dissolved for about 2-3 minutes.

In a 10 mL vial containing 2 mg of pyrazineamide, 100 μL of a formamidine sulfinic acid solution was added using a micropipette to generate a first mixture. The first mixture is sealed with an n-butyl rubber cap and then sealed with aluminum to maintain a precursor state that is easy to associate with the radionuclide.

2. Mixing Step

About 1 mL of sodium pertechnetate solution containing radionuclide Tc-99m (Technetium-99m) was injected into the first mixture by using a syringe to inject the first mixture and sodium pertechnetate solution. To produce a second mixture that is a mixture. At this time, Tc-99m (Technetium-99m) per 1 mL of the second mixture is 40 mCi.

3. Joining step

The second mixture was placed in a screw-type plastic container and heated for 12 seconds in a 1000 W microwave oven and 15 seconds in an 800 W microwave oven, resulting in the formation of pyrazineamide and radionuclide Tc-99m (Technetium-99m). A pulmonary tuberculosis nuclear medicine imaging kit was prepared. Immediately remove the heated kit from the microwave and place it in a shielded area, allow it to stand at room temperature for about 5 minutes, add water for injection into the kit, and filter with a sterile filter (a disc type with a pore size of 0.22 μm). After that, Tc-99m (Technetium-99m) formed a complex containing pyrazine amide and a chelate ring in a sterile vacuum bottle.

When the image contrast agent is injected intravenously by the kit, the infection site of pulmonary tuberculosis bacteria can be found through gamma camera photography. The contrast agent comprises an aqueous saline medium and can be administered via intravenous injection. The medium may, of course, include commonly used pharmaceutical adjuvants such as pharmaceutically acceptable salts, buffers or preservatives.

As mentioned above, although the manufacturing method of the kit which concerns on this invention was demonstrated to one Example, this invention is not limited to this. For example, 40 m Ci of sodium pertechnetate containing Tc-99m (Technetium-99m) is added to the first mixture in the mixing step, but the amount of sodium pertechnetate is 30 Can be ˜50 mCi and add other compounds comprising the radionuclide Tc-99m (Technetium-99m) or include other nuclides, such as Tc-99m, Indium-111, Iodine-131, Iodine-123 Compounds may be added to cause radioactive release.

Hereinafter, a method for diagnosing pulmonary tuberculosis infection using a kit according to the present invention will be described. Pulmonary tuberculosis infection diagnosis method using a kit according to the present invention includes an injection step, a photographing step and a judgment step.

1. Injection step

An imaging contrast agent using the kit prepared in Example 1 is injected intravenously into an animal or human body. Intravenous dose is about 10 mCi / 5 ~ 6 mL per person.

2. Shooting Step

After a certain period of time has passed since the injection phase, animals or patients who have been injected intravenously with a gamma camera are photographed. In this case, the radioactive material invented emits radioactivity, and the radioactivity is detected in a large amount at the pulmonary tuberculosis infection site as photographed through a gamma camera. There are two stages: immediate shooting, which is taken immediately after intravenous injection, and delayed shooting, which is taken after a considerable time has elapsed. Immediate shooting is suitable for about 1 hour and delayed shooting for 4 hours. In the case of long-term shooting, the shooting time may be one or several days. Radioisotopes with short half-lives, such as Tc-99m or compounds containing them, are suitable for single shots within 6 hours, and radioactive isotopes with long half-lives, for example, when taking multiple shots over 6 hours or days. For example, Indium-111, Iodine-131, Iodine-123 or a compound containing them are suitable.

3. Judgment

Pulmonary tuberculosis infection is determined by whether radioactivity is recorded on the captured image and the concentration of radioactivity.

Figure 3 is an image taken to take the human body, can check the pulmonary tuberculosis generated in the lower right lung. In detail, FIG. 3 is a photograph showing the patient's chest in transverse, sagittal, and coronal planes. similar). The three axes of the pictures shown are composed of red targets corresponding to each other, so that the lesions (arrows) in the lower right lung can be imaged black. FIG. 4 is a corresponding X-ray photograph, and the image contrast agent is administered according to the present invention. Confirmed lower right lung lesions (arrows) are shown.

The present invention provides a tuberculosis infection nuclear medicine imaging agent and a kit (kit) in which radionuclides are combined with pyrazine amide, and thus, it is possible to accurately determine whether the tuberculosis is active or infected. In addition, the present invention can identify the progress of treatment according to the use of anti-tuberculosis drugs, there is an advantage that can prevent overdose or insufficient treatment of the drug.

Claims (4)

Imaging of active tuberculosis infections comprising a stable complex of radionuclides selected from the group consisting of pyrazineamide of Formula 1 or a pharmaceutically acceptable salt thereof and Tc-99m, Indium-111, Iodine-131 and Iodine-123 Pharmaceutical composition for contrast agent.

A pyrazineamide of Formula 1 or a pharmaceutically acceptable salt thereof is included together with a reducing agent to form a complex with a radionuclide selected from the group consisting of Tc-99m, Indium-111, Iodine-131 and Iodine-123, Kit for use in the preparation of active TB infection imaging contrast agents.

The kit of claim 2, wherein said reducing agent is formamidine sulfinic acid or formamidine sulfonic acid. The kit of claim 2 or 3, wherein the mass ratio of pyrazineamide and reducing agent is 2: 1.

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