CN1995020B - Radioactive iodine (*I)-17-propylamino-17-desmethoxygeldanamycin and its preparation method - Google Patents

Abstract

The preparation method of radioactive iodine ( ^* I)-17-propylamino-17-desmethoxygeldanamycin relates to the technical fields of nuclear medicine, oncology and pharmaceutical preparations. The compound iodine ( ^* I) 17-propylamino-17-desmethoxygeldanamycin of the present invention is an inhibitor of HSP90 (heat shock protein 90) 17-propenylamino-17-desmethoxygeldanamycin. Iodine-labeled product of dermycin. The preparation method is to modify the structure of 17-propenylamino-17-desmethoxygeldanamycin, and introduce radionuclide iodine into the double bond at its terminal. The present invention also provides a drug or a pharmaceutical composition comprising iodine-17-propylamino-17-desmethoxygeldanamycin as a pharmaceutically acceptable carrier. The drug or pharmaceutical composition can be used for various Tumor/cancer treatment and diagnosis. The compound of the present invention is reported for the first time. Its preparation process is simple and its purity is high. It not only has the targeting property of HSP90 inhibitors, but also combines the advantages of radioactivity and chemotherapeutic drugs, which can enhance the therapeutic effect of tumor/cancer; at the same time, it is also convenient for clinicians. Get tumor diagnostic reagents at any time, and provide patients with non-invasive and timely SPECT/PET imaging.

Description

Radioactive iodine (*I)-17-propylamino-17-desmethoxygeldanamycin and its preparation method

technical field

The invention relates to a tumor treatment reagent radioactive iodine ( ^* I)-17-propylamino-17-desmethoxygeldanamycin, a tumor diagnostic agent (single photon emission computed tomography/SPECT, positron Emission computed tomography/PET) and preparation method and application of medicine or medicine composition thereof. It involves the technical fields of nuclear medicine, oncology, and pharmaceutical preparations.

Background technique

Malignant tumors are diseases that seriously threaten human survival and health, and early diagnosis is difficult. In recent years, the morbidity and mortality of tumors have been increasing year by year. The loss of surgical opportunities and recurrence and metastasis after surgery are the main reasons that make tumors difficult to control and accelerate the death of patients. At present, the treatment of tumors mainly relies on comprehensive measures such as surgery, chemotherapy, radiotherapy and biological therapy. However, although the short-term effect of these conventional treatment methods on the prevention and treatment of tumor metastasis is good, the long-term effect is poor. The development of tumor nuclear medicine provides new ideas and methods for the early diagnosis and treatment of tumors. As one of the comprehensive treatment methods, the application of radionuclide internal radiation therapy in the treatment of solid tumors has been paid more and more attention in recent years. According to the theory of radiation biology, the therapeutic effect of radionuclides is persistent; the sensitivity of cells to nuclear radiation is related to the functional state of cells, and malignant tumor cells proliferate rapidly and are more sensitive to nuclear radiation. Therefore, the number of courses of radionuclide internal radiation therapy is small, and good and long-lasting effects can be achieved.

The principle of radionuclide internal radiation therapy is to deliver appropriate radiopharmaceuticals with high energy transmission line density to the lesion or its surroundings. The nuclear radiation can cause denaturation and necrosis of tumor tissues within the effective range. Radiological injury to adjacent normal tissue outside the range of the radiation. Radiopharmaceuticals use molecules or ions that specifically act on diseased tissues as carriers of radionuclides, which can make radionuclides mainly concentrate in tumor tissues after administration, and stay in tumor tissues for a long time. The accumulation of radionuclides in normal tissues and organs other than normal tissues should be reduced as much as possible. Therefore, it is very suitable for local high-dose internal irradiation treatment of tumors, and can minimize the damage of radiation to adjacent important tissues and organs.

β-rays are superior to γ-rays in terms of tissue dose attenuation and radiation protection, so the use of β-rays to treat tumors has become a hot spot in radiopharmaceutical research. Radionuclides used for medical purposes must be able to emit radiation with high energy transmission linear density to destroy abnormally proliferating and differentiated tissues. ¹³¹ I is the most widely used imaging and therapeutic radiopharmaceutical, used for thyroid function measurement and imaging, treatment of thyroid cancer and hyperthyroidism; ¹²³ I, ¹²⁴ I are used for SPECT and PET imaging respectively.

With the in-depth research on the pathogenesis of tumors at the molecular level, researchers have found that heat shock protein 90 (Heat-Shock Proteins 90, HSP90), which acts as a molecular chaperone in vivo, plays an important role in the development of tumors. The research on inhibitors of the target is in the ascendant. The effector proteins of HSP90 in the body are often protein kinases and transcription factors that play an important role in the body's signal transduction pathways, including tyrosine kinase receptor (Her-2), vascular endothelial growth factor receptor (EGFR), cell cycle CDK4, mutant p53, and hypoxia-inducible factor 1α (HIF-1α) play a role in the blood supply of ovarian cancer, breast cancer, prostate cancer, gastric cancer, retinoblastoma and kidney cancer, etc. A protein that plays an important role in nutritional support, signal stimulation, and substance metabolism. HSP90 inhibitors can effectively inhibit multiple cell signal transduction pathways by inhibiting HSP90, and treat breast cancer, prostate cancer, melanoma, ovarian cancer, cervical cancer, liver cancer, lung cancer, thyroid cancer, leukemia, etc.

17-allylamino-17-demethoxygeldanamycin (17-allylamino, 17-demethoxygeldanamycin, 17-AAG) is the first HSP90 inhibitor to enter clinical trials. -AAG has high affinity. Studies have shown that the affinity between 17-AAG and HSP90 derived from tumor cells is 20-200 times that of HSP90 derived from normal tissues. Therefore, 17-AAG can be used as a suitable carrier to carry radionuclides for diagnosis, Treat tumors. 17-AAG is a purple-like powder with a molecular weight of 585.7. It is easily soluble in methanol, DMSO, chloroform, ethyl acetate, ethanol and other organic solvents. It is insoluble in water and becomes a lavender clear liquid after dissolution. 17-AAG was obtained from Sigma Company. Its molecular formula is C ₃₁ H ₄₃ N ₃ O ₈ , and its structural formula is:

17-AAG can inhibit multiple cell signal transduction pathways, making the effector proteins of HSP90, including ERB-B2, HER2, estrogen receptor, androgen receptor, RAF, P13 kinase signaling protein AKT, chronic myelogenous leukemia ( CML) proliferation fusion proteins BCR-ABL, CDK4, h-TERT, etc. are degraded through the ubiquitinated proteasome pathway; at the same time, they affect multiple cell signal transduction pathways to achieve multi-point attack, destroying the entire signal pathway network on which tumors depend for survival, so that Tumor cell apoptosis and growth were inhibited. 17-AAG can induce leukocyte aggregation, inhibit the growth of tumor cells in the G2 phase, increase the expression of E-cadherin, change wild-type and mutant estrogen and androgen receptors, inhibit their transcriptional activity, and eliminate hormones that promote ligand dependence Receptor activates multiple cellular signaling pathways. A large number of studies have shown that 17-AAG increases the sensitivity of tumors to chemotherapy drugs, reverses the sensitivity of drug-resistant tumors to drugs, and increases the sensitivity of tumor cells to radiation (such as using 17-AAG to induce cancer cells first, and then using ionizing radiation, found that The sensitivity of cancer cells to ionizing radiation increases after 6-48 hours of administration). Therefore, 17-AAG can be used to treat breast cancer, prostate cancer, melanoma, ovarian cancer, cervical cancer, CML, lung cancer, liver cancer, thyroid cancer, etc.

Contents of the invention

Technical problem: the purpose of this invention is to provide a kind of radioactive iodine ( ^* I)-17-propylamino-17-demethoxygeldanamycin and its preparation method. The preparation process of the present invention is simple, the markers have high purity and good stability, can be used for diagnosis in small doses, and can be used for treating tumors in large doses, so that patients can obtain non-invasive imaging and treatment at any time.

Technical scheme: technical scheme of the present invention comprises two contents:

(1) A preparation method of ^* 1-17-propylamino-17-desmethoxygeldanamycin;

(2) A pharmaceutical composition with ^* 1-17-propylamino-17-desmethoxygeldanamycin as an active ingredient.

^{The 123} I compound of the present invention can be used in SPECT (single photon emission computed tomography), and the ¹²⁴ I compound can be used in PET (positron emission computed tomography), which is a unique non-invasive, living, and dynamic imaging method in nuclear medicine.

^{The *} I-17-propylamino-17-desmethoxygeldanamycin of the present invention can be used as an active ingredient together with a pharmaceutically acceptable excipient to prepare a pharmaceutical composition, and the pharmaceutical composition can be It is prepared into various dosage forms by conventional methods in the preparation field, such as injection, tablet, powder, granule, pill, capsule, tincture, oral liquid, ointment, cream, emulsion or application. According to different dosage forms, the excipients used in the pharmaceutical composition are also different, commonly used excipients include diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants , Adsorption carrier, lubricant, emulsifier, osmotic pressure regulator.

^* I-17-propylamino-17-desmethoxygeldanamycin involved in the present invention is prepared through the following steps:

1.) Mix the ethanol solution of 17-propenylamino-17-desmethoxygeldanamycin with the oxidizing agent and Na ^* I under acidic conditions, and place it at 20-60°C for 10-30min;

2.) Add a reducing agent to the mixture in step 1) to terminate the reaction; add a pH regulator to adjust the pH to neutral;

3.) Add organic phase extraction to the product of step 2), take the organic layer, and dry it with nitrogen;

The acidic condition in the above-mentioned step 1) can be selected from one of hydrochloric acid, acetic acid and sulfuric acid solution.

There are many methods of iodine labeling, such as chloramine T method, iodine chloride method (ICl), Iodogen method, peroxide oxidation method and so on. The present invention adopts the peroxide oxidation method, which has high marking rate and few impurities. The oxidant selected can be one of hydrogen peroxide and peracetic acid.

The reductant in above-mentioned step 2) can select a kind of sodium metabisulfite, sodium thiosulfate for use. The reagent for adjusting pH should be selected from one of sodium hydroxide, phosphate buffer, sodium carbonate/sodium bicarbonate buffer, and ammonia water.

The organic phase in the above step 3) can be selected from one of ethyl acetate, ethanol, ether, acetone, and acetonitrile.

The preferred conditions are as follows:

A: Dissolve 20 μg of 17-propenylamino-17-desmethoxygeldanamycin in 100 μL of ethanol, add 20 μL of 4mol/L hydrochloric acid solution, add 20 μL of 7% hydrogen peroxide, and add 37 to 370 MBq (1 to 10 mCi) of Na ^* I , vortexed for 1min to mix well, and placed at 40°C for 15min; adding 0.01mol/L sodium metabisulfite to terminate the reaction. Adjust the pH to 7 with ammonia water.

B: Add 2 mL of water to A, extract with ethyl acetate (3 mL x 3 times), collect the organic phase, and blow dry with nitrogen to obtain the final product.

The ^* I-17-propylamino-17-desmethoxygeldanamycin and its pharmaceutical composition of the present invention are beneficial to the treatment, diagnosis and prevention of various tumors.

17-Propylamino-17-desmethoxygeldanamycin has strong fat solubility and low polarity. The present invention intends to modify the structure of 17-propenylamino-17-desmethoxygeldanamycin, introduce radionuclide iodine into its double bond, enhance the polarity and targeting of the compound, and better use in the diagnosis and treatment of tumors. The present invention makes a bold attempt to develop the active ingredients of existing antineoplastic drugs into radionuclide tumor diagnosis and treatment drugs, and organically combines HSP90 inhibitors with nuclides, so that it can be applied to the localization diagnosis of tumors and their metastases in small doses , it can be applied to the treatment of tumors in large doses. The present invention provides a new idea for the development of new radioactive drugs; the pro-tumor property of the drug itself combined with the radioactive radiation of the nuclide can better exert the targeting of anti-tumor drug treatment, reduce toxic side effects and damage to normal tissues; at the same time Provide tumor patients with new and effective reagents for non-invasive imaging, which is helpful for early diagnosis and treatment of tumors.

The present invention is also applicable to 17-AAG and other compounds with structures similar to 17-AAG.

Beneficial effect: From the above results, the advantages of the present invention can be drawn as follows.

(1) The present invention has prepared a new compound ^* I-17-propylamino-17-desmethoxygeldanamycin.

(2) ^* I-17-propylamino-17-desmethoxygeldanamycin of the present invention is safe and non-toxic, retains the original biological characteristics of 17-AAG, and has a targeting effect in tumors Strong and synergistic effect, indicating a good medicinal prospect.

(3) The preparation process of the present invention is simple, the markers have high purity and good stability, and can be used for tumor diagnosis in small doses and for tumor treatment in high doses, so that patients can obtain non-invasive imaging and treatment at any time.

(4) The present invention ^* I-17-propylamino-17-desmethoxygeldanamycin can be used as an active ingredient together with a pharmaceutically acceptable excipient to prepare a pharmaceutical composition, and can be used for Treatment of various tumors.

(5) The present invention opens up a new application field, which can be used for tumor treatment, SPECT and PET imaging.

Detailed ways

Embodiments of the present invention will be described below, but the content of the present invention is not limited thereto at all. The iodide of geldanamycin can completely adopt this method.

Example

(1): ^* Preparation of I-17-propylamino-17-desmethoxygeldanamycin

A: Dissolve 20 μg of 17-propenylamino-17-desmethoxygeldanamycin in 100 μL of ethanol, add 20 μL of 4mol/L hydrochloric acid solution, add 20 μL of 7% hydrogen peroxide, 37～370MBq (1～10mCi) Na ^* I, Vortex for 1 min to mix evenly, and place at 40°C for 15 min; add 0.01 mol/L sodium metabisulfite to terminate the reaction. Adjust the pH to 7 with ammonia water.

B: Add 2 mL of water to A, extract with ethyl acetate (3 mL x 3 times), collect the organic phase, blow dry with nitrogen, and obtain the product.

Iodine ( ^* I) in Na ^* I is a kind of in nuclide iodine ^123I , ^124I , ^125I , ^127I , ^131I ; Oxidizing agent is hydrogen peroxide, a kind of in peracetic acid; Acid condition is hydrochloric acid, One of acetic acid and sulfuric acid; the reducing agent is one of sodium metabisulfite and sodium thiosulfate; the pH regulator is one of sodium hydroxide, ammonia water, sodium carbonate/sodium bicarbonate buffer, and phosphate buffer ; The organic phase of extraction is a kind of in ethyl acetate, ether, ethanol, acetone, acetonitrile.

(2): ^* Identification of I-17-propylamino-17-desmethoxygeldanamycin

Determination by thin layer chromatography and high performance liquid chromatography.

A: Thin-layer chromatography: thin-layer silica gel GF254 plate, polyamide film or silica gel paper as the support, the developing solvent is dichloromethane/methanol=9/1, Rf=0.6-0.9.

After the sample is applied, the development is completed, and the markers are detected as light yellow spots at Rf = 0.6-0.9 under ultraviolet light; the chromatography paper is cut into ten sections, and the radioactive count rate is measured by a gamma counter, and at Rf = 0.6-0.9 Concentration of radioactivity, free iodine at Rf = 0 ~ 0.1.

B: HPLC: BIORAD HPLC, YWG C18 reverse-phase column, 50% acetonitrile plus 25mM phosphate (pH 3.00) containing 10mM triethylamine, flow rate 1mL/min, simultaneously detected by ultraviolet detector (wavelength 334nm) and radioactive detector, The retention time of free iodine is about 1.3min, ^* The retention time of I-17-propylamino-17-desmethoxygeldanamycin is about 12min.

(3): ^* Stability of I-17-propylamino-17-desmethoxygeldanamycin

Take ^{the *} I-17-propylamino-17-desmethoxygeldanamycin obtained by drying under the item of Example (1), add physiological saline to dissolve, and put it in a refrigerator at 4°C. After standing for 8 days, the radiochemical purity is still greater than 90%. It shows that the marker is stable at 4°C and can meet the requirements of routine clinical use.

Claims (4)

1. a preparation method of radioactive iodine ( ^* 1)-17-propylamino-17-demethoxygeldanamycin, characterized in that, 17-propenylamino-17-demethoxy Geldanamycin is radioactively iodine-labeled by peroxide oxidation method, including the following steps: 1.) Mix the ethanol solution of 17-propenylamino-17-desmethoxygeldanamycin with the oxidizing agent and NaI ^* under acidic conditions, and place it at 20-60°C for 10-30min; 2.) Add a reducing agent to the mixture in step 1) to terminate the reaction; add a pH regulator to adjust the pH to neutral; 3.) Add organic phase to the product of step 2) for extraction, take the organic layer, and dry it. 2. the preparation method of radioactive iodine ( ^* 1)-17-propylamino group-17-demethoxy geldanamycin according to claim 1, is characterized in that step 1) described oxygenant is hydrogen peroxide , a kind of in peracetic acid; Described acidic condition is a kind of in hydrochloric acid, acetic acid, sulfuric acid. 3. the preparation method of radioactive iodine ( ^* 1)-17-propylamino-17-demethoxygeldanamycin according to claim 1, is characterized in that step 2) described reducing agent is One of sodium metabisulfite and sodium thiosulfate; the pH regulator is one of sodium hydroxide, ammonia water, sodium carbonate/sodium bicarbonate buffer, and phosphate buffer. 4. the preparation method of radioactive iodine ( ^* 1)-17-propylamino-17-demethoxygeldanamycin according to claim 1 is characterized in that the organic phase described in step 3) is One of ethyl acetate, ether, ethanol, acetone, and acetonitrile.