CN101200448B - β-Elemene Derivatives and Their Re Complexes and 188Re Labeled Objects - Google Patents

Abstract

The invention discloses β-elemene dipyridylamine derivatives of the compound of the formula I and its Re complex-the compound of the formula II and ^{the 188} Re label-the compound of the formula III, as well as their synthesis methods. Various compounds of the invention have anticancer activity and lower toxic and side effects, and are potential anticancer drugs.

Description

β-Elemene Derivatives and Their Re Complexes and 188Re Labeled Objects

technical field

The present invention relates to a derivative of β-elemene, especially β-elemene dipyridylamine derivatives and their Re complexes and radionuclide ¹⁸⁸ Re markers, and the synthesis method of these compounds .

Background technique

Nuclear medicine can use the special physical properties of labeled nuclides in some radiopharmaceuticals to perform internal radiation therapy while imaging a variety of tumors and their lesions. This unique method of tumor diagnosis and treatment has been increasingly accepted in clinical practice. Pay attention to. At present, such radionuclides with both therapeutic and imaging functions mainly include ¹³¹ I, ¹⁵³ Sm, ^117m Sn, ¹⁸⁶ Re, ¹⁸⁸ Re, etc. Among them, ¹⁸⁸ Re has attracted much attention in the field of tumor diagnosis and treatment because of its excellent physical and chemical properties.

The half-life of ¹⁸⁸ Re is 16.9 hours, and the main β-energy is 2.12MeV. It has a short range in the tissue (more than 95% is absorbed within 4mm), and has little damage to the surrounding tissue, accompanied by a branch ratio of 15 % of 155keV gamma rays are suitable for imaging, which is convenient for clinically estimating absorbed dose and conducting pharmacokinetic research. Moreover, the Na ¹⁸⁸ ReO ₄ solution can be easily obtained from the ¹⁸⁸ W- ¹⁸⁸ Re generator at any time. The biological behavior of ¹⁸⁸ Re-sodium perrhenate (free form of ¹⁸⁸ Re) is also very good. Studies have shown that after intravenous injection of ¹⁸⁸ Re-sodium perrhenate in experimental animals, ¹⁸⁸ Re-sodium perrhenate has only a small amount of organ damage. It is absorbed and quickly excreted through urine, and its biological half-life in the human body is less than 10 hours, so even if ¹⁸⁸ Re-labeled radiopharmaceuticals undergo nuclide shedding in the body, it will not cause serious radiation damage to the human body ^[1] . The excellent nuclear properties of ¹⁸⁸ Re and the widespread application of ¹⁸⁸ W- ¹⁸⁸ Re generators have made ¹⁸⁸ Re radiopharmaceuticals a research hotspot in the field of radiopharmaceuticals.

In recent years, a great breakthrough has been made in the synthesis of kinetically inert low-valent rhenium(I) compounds [ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ^{+ [2]} , thus providing a new opportunity for the development of ¹⁸⁸ Re radiopharmaceuticals. Development has injected new vitality. However, [ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ cannot be used as a radiopharmaceutical alone, and when it is used to directly label biomolecules, the labeling rate is very low, and the labeled compounds are very unstable. Therefore, when labeling biomolecules, bifunctional chelating agents (bifunctional chelating agents, BFCA) are required in the middle. BFCA is like a bridge, one end is connected to the target compound to be labeled, as a targeting molecule or for adjuvant therapy, and the other end is complexed with radionuclide ¹⁸⁸ Re. BFCA molecules not only have strong chelating groups, but also contain groups that can form covalent linkages with targeting molecules, biomacromolecules, monoclonal antibody proteins, etc., that is, biomolecules are bound to [ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ forms a strong bond.

β-Elemene (β-Elemene) is a kind of volatile oil-like natural compound extracted from the rhizome of Zingiberaceae Wen Curcuma (Ezhu) in China in recent years. It has anti-cancer effect. Its molecular formula is C ₁₅ H ₂₄ , and its structural formula as follows:

The nuclide ¹⁸⁸ Re can kill tumor cells, and the ligand in the ¹⁸⁸ Re label can guide the nuclide to reach the tumor cells. The radionuclide will inevitably cause damage to normal tissues while killing tumor cells, while β-olium Argan has the functions of radiation sensitization and improving human immunity. Therefore, it is hoped that the synthesized β-elemene ¹⁸⁸ Re complex has lower toxicity and side effects. But so far, there are few reports on the labeling of natural products with ¹⁸⁸ Re nuclides. This is because the structures of natural products are complex. Functional chelating agent, and the separation, purification and structural modification of natural products are quite difficult.

Contents of the invention

The technical problem to be solved in the present invention is the above-mentioned subject, and a kind of β-elemene derivative that coordinates as metal ¹⁸⁸ Re is provided—— ¹⁸⁸ Re β-elemene dipyridylamine derivatives, as a structural reference The β-elemene metal rhenium (Re) complex and β-elemene dipyridylamine derivatives, and their synthesis methods.

For this reason, the present invention screens out the bifunctional chelating agent that can not only be chelated with Re, but also be connected with β-elemene——dipicolylamine derivatives, and at first synthesized the β-elemene that can chelate Re Elemene derivatives, that is, β-elemene dipyridylamine derivatives, its structural formula is shown in the following formula I; then synthesized β-elemene metal rhenium complex: β-elemene dipyridylmethyl The Re(CO) ₃ organic complex of amine derivatives, its structural formula is shown in the following formula II, which is a cold experiment of radiosynthesis; finally synthesized ¹⁸⁸ Re-labeled β-elemene dipyridylamine derivatives , whose structural formula is shown in the following formula III.

where A represents -CH ₂ -, or

CH₂CH₂O

_m

CH₂

_n其中m＝0-2，n＝1-6。-NH

_{CH2CH2O _}

_m

CH ₂

_n where m=0-2, n=1-6.

Preferably, said m=0, n=2 or 6; or m=2, n=2.

Preferred compounds of formula I of the present invention are β-elemene dipyridylmethylamine, β-elemene monosubstituted 2-[bis(2-pyridylmethyl)-amino]-ethylamine, β-elemene -6-[bis(2-pyridylmethyl)-amino]-hexylamine and N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}- Ethyl-N'-β-elemeneamine, the compound of formula II is β-elemene dipyridylmethylamine tricarbonyl rhenium complex, β-elemene monosubstituted 2-[bis(2-pyridinemethyl Base)-amino]-ethylamine tricarbonyl rhenium complex, β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine tricarbonyl rhenium complex and N-2 -{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemeneamine tricarbonyl rhenium complex, the compound of formula III is ¹⁸⁸ Re-labeled β-elemene dipyridylamine, ¹⁸⁸ Re-labeled β-elemene monosubstituted 2-[bis(2-pyridylmethyl)-amino]-ethylamine, ¹⁸⁸ Re-labeled β -Elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine and ¹⁸⁸ Re-labeled N-2-{2-[2-(bis-2-pyridine-methylamine)- Ethoxy]-ethoxy}-ethyl-N'-β-elemenamine.

The synthesis method of the compound of formula I of the present invention comprises reacting chlorinated β-elemene (mainly compound of formula IV) and dipyridylamine derivatives (compound of formula V) in an acetonitrile solvent. Its reaction scheme is as follows.

where ^R1 is hydrogen, or

_m CH_{2 n}其中m、n如上定义。NH _{2 CH2CH2O _}

_m CH _{2 n} wherein m and n are as defined above.

Preferably, the molar ratio of the chlorinated β-elemene to the compound of formula V is 0.1-10:1, and the molar ratio of the compound of formula V is 1-12.5:1 equivalent basic catalyst, and the reaction temperature is 0 -80°C, the reaction time is 1-48 hours.

The present invention prepares pure monosubstituted β-elemene derivatives by carefully adjusting the molar ratio of reactants, reaction temperature and other conditions.

The molar ratio of the above-mentioned chloro-beta-elemene to the compound of formula V is more preferably 1-3: 1, the molar ratio of the basic catalyst to the compound of formula V is more preferably 1-3: 1, and the reaction temperature is more preferably 60 to 70°C. The time is more preferably 8 to 21 hours.

The basic catalyst may preferably be sodium hydroxide.

More preferably, KI can also be added to the above reaction system of the present invention, and KI can undergo a displacement reaction with chloro-beta-elemene, so that the reaction speed between chloro-beta-elemene and the compound of formula V is faster. Its dosage is about 5%-30% (mole percentage, compared with dipyridylamine), preferably 10-15%.

The synthesis method of the compound of formula II in the present invention comprises reacting the compound of formula I with rhenium tricarbonyl in methanol solution. Its reaction scheme is as follows.

Among them, rhenium tricarbonyl——[N(Et) ₄ ] ₂ [Re(CO) ₃ Br ₃ ] (tetraethylammonium bromide tricarbonyl rhenium complex) can be synthesized using existing techniques (Alberto R, Egli A, Abrum U, et al, Synthesis and reactivity of[Net ₄ ] ₂ [ReBr ₃ (CO) ₃ ]Formation and Structural Characterization of the Clusters[Net ₄ ][Re(μ ₃ -OH)(μ-OH) ₃ (CO) ₃ ] and [Net ₄ ][Re ₂ (μ-OH) ₃ (CO) ₆ ] by Alkaline Titration J Chem Soc Dalton Trans, 1994, 2815-2820.). And [N(Et) ₄ ] ₂ [Re(CO) ₃ Br ₃ ] and the synthetic method of formula I compound also can refer to prior art (such as above-mentioned document or Schibli R, Schwarzbach R, Alberto R, et al.Steps Toward High Specific Activity Labeling of Biomolecules for Therapeutic Application: Preparation of Precursor[ ¹⁸⁸ Re(H ₂ O) ₃ (CO) ₃ ] ⁺ and Synthesis of Tailor-Made Bifunctional LigandSystems. Bioconjugate Chem, 2002, 13(4): 750-756.) .

Preferably, the molar ratio of the compound of formula I to rhenium tricarbonyl is about 1:1, the reaction temperature is room temperature (about 25° C.) to 60° C., and the reaction time is 0.5 to 1 hour.

The method for synthesizing the compound of formula III of the present invention comprises radioactively labeling the compound of formula I with Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ .

Existing radioactive labeling techniques are all carried out in NaCl aqueous solution. The water solubility of elemene in the present invention is not good, so the reaction conditions are adjusted during labeling, and organic solvents such as ethanol are added to help dissolve, so that the labeling rate is greatly improved. Therefore, preferably, the radiolabeling includes adding the Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ solution to the ethanol solution of the compound of formula I, using an incubator with a temperature control of 75°C and a rotation speed of 1300 rev/min, the reaction time was 50 minutes, and then separated by HPLC.

The present invention synthesizes the ligand of Re - the compound of formula I; successfully synthesized the compound of formula II of the β-elemene metal rhenium (Re) complex, that is, the Re of β-elemene dipyridylmethylamine and its derivatives (CO) ₃ organic complexes; and finally obtained β-elemene dipyridylmethylamine and its derivatives labeled with formula III compound ¹⁸⁸ Re. Fragrantene. These compounds have anticancer activity and are a potential anticancer drug.

Description of drawings

Fig. 1 is the HPLC spectrogram of the non-radioactive β-elemene dipyridylamine Re(CO) ₃ complex of the present invention, t=20.75min.

Fig. 2 is the HPLC spectrogram of radioactive β-elemene dipyridylamine Re(CO) ₃ complex, t=21.23min.

Fig. 3 is the HPLC spectrogram of the β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine tricarbonyl rhenium complex of the present invention, t=21.175min.

Fig. 4 is the HPLC spectrogram of the radioactive β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine tricarbonyl rhenium complex, t=21.21min.

Figure 5 is the tricarbonyl rhenium complex of N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemeneamine HPLC spectrum, t=21.187min.

Figure 6 shows the radioactive N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemeneamine tricarbonyl rhenium complex The product HPLC spectrogram, t=21.190min.

Detailed ways

The present invention is further illustrated below with examples, but the present invention is not limited thereto.

The raw material chlorinated β-elemene used in the following examples can be synthesized according to the prior art, such as the various documents mentioned in the above-mentioned background technology content are synthesized by themselves; company's product.

1. the synthesis of formula IV compound chlorinated β-elemene

Synthesis of β-elemene chlorides according to the literature (Jia Weimin, Synthesis, structure and structure-activity research of new anticancer drug β-elemene and its derivatives, doctoral thesis of Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 1991), the specific steps are as follows : Dissolve 0.01mol β-elemene in 10mL dichloromethane, add 2mL formic acid, control the temperature at 0-5°C, slowly add 15mL sodium hypochlorite solution dropwise within 2 hours, and continue the reaction for 3-5 hours. After the reaction is complete, add saturated aqueous sodium bicarbonate solution to adjust the pH to 7-8, separate the reaction mixture and separate the organic phase. Sodium drying, filtration, removal of solvent dichloromethane, a light yellow oily substance, a colorless oily substance can be obtained after silica gel column chromatography, which is a mixture of chlorinated β-elemenes by gas chromatography, which is β-elemene A mixture of alkenyl chloride and β-elemene, the content of β-elemene chloride is about 40-50%. Wherein the β-elemene chloride contains the following three compounds, mainly monosubstituted, i.e. formula IV and formula IV' compounds, wherein the formula IV compounds are the main compounds, and the content of formula IV' compounds is very small.

Because the β-elemene in this chlorinated β-elemene mixture is not easy to separate, and after the reaction finishes, because the polarity of β-elemene is very different from the polarity of the final product of the present invention, it is very easy to separate . Therefore, similar to the prior art, the chlorinated β-elemene mixture can be directly used in the synthesis reaction of the present invention, and the compound participating in the reaction is mainly the compound of formula IV.

The synthesis of embodiment 1 β-elemene dippicolylamine:

0.8110g (4.0mmol) dipyridylamine (compound a), 0.1646g (4.1mmol) NaOH, 1.8027g chlorinated β-elemene mixture (7.6mmol), 0.0512g (0.3mmol) KI, 8ml CH ₃ CN . Stir magnetically under an oil bath at 63°C, heat for 21 hours, the solution turns brown, add NaCl solution after the reaction, and then extract with CHCl ₃ 3×10ml to obtain a dark brown oily liquid.

When the developer is ethyl acetate/CH ₃ OH (volume ratio is 10:1), the Rf value of the product is 0.59, and it passes through a silica gel column [silica gel: 200-300 mesh, product of China Pharmaceutical (Group) Shanghai Chemical Reagent Co., Ltd., washed Dehydration: ethyl acetate] purification to obtain 1.05 g of the product β-elemene dipyridylmethylamine (compound 1) as a yellow liquid with a yield of 64%.

Its identification data are as follows:

IR V _max : 3080 (C=CH), 1640 (C=C).

¹ H-NMR (CDCl ₃ , TMS, 500MHz), δ0.90(3H, s), 1.47-1.27(6H, m), 1.61(3H, s), 1.92-1.89(1H, m), 2.07-2.04 (1H, m), 3.06 (2H, q, J = 14.0Hz), 3.73 (4H, q, J = 14..1Hz), 4.47 (1H, s), 4.73 (1H, s), 4.84 (1H, dd, J=5.82Hz, 1.08Hz), 4.85(1H, d, J=1.3Hz), 4.89(1H, s), 5.06(1H, s), 5.77(1H, dd, J=10.48Hz, 7.38Hz ), 7.09-7.07 (2H, m), 7.50-7.47 (2H, m), 7.60 (2H, dt, J=7.65Hz, 1.57Hz), 8.45 (2H, d, J=4.3Hz).

¹³ C-NMR (CDCl ₃ , TMS, 125MHz), δ17.39, 25.31, 27.68, 33.73, 40.57, 40.74, 42.56, 53.74, 59.71, 60.80, 110.53, 111.63, 112.71, 122.62, 123.41, 1147.16, 4 , 150.94, 152.06, 160.46.

MS [M ⁺ ] = 401, HRMS, calculated 401.2831, found 401.2842.

Example 2 Synthesis of β-elemene-2-[two (2-pyridylmethyl)-amino]-ethylamine

Dissolve 3.3mmol compound b, 4mmol NaOH, 1.8027g (7.6mmol) chlorinated β-elemene mixture, 0.0512g (0.3mmol) KI in 8ml CH ₃ CN, stir magnetically in an oil bath at 63°C, and heat for 21 hours , the solution was brown. After the reaction, NaCl solution was added, and then extracted with CHCl ₃ (3×10ml) to obtain a dark brown oily liquid.

When the developer is ethyl acetate/CH ₃ OH, the Rf value of the product is 0.59, and the product is purified by silica gel column to obtain the product β-elemene monosubstituted 2-[bis(2-pyridylmethyl)-amino]-ethyl Amine (compound 2), yield 74%, identification result is as follows:

¹ H-NMR (CDCl ₃ , TMS, 500MHz), δ1.00(s, 3H), 1.38-1.77(m, 6H), 1.69(s, 3H), 1.97-2.02(m, 1H), 2.10-2.23 (m, 1H), 3.22(s, 4H), 3.71(s, 2H), 3.95(s, 4H), 4.56(s, 1H), 4.77(s, 1H), 4.89(d, J=2.18, 1H ), 4.91 (d, J=8.42, 1H), 5.30 (s, 1H), 5.35 (s, 1H), 5.79 (dd, J=17.72, 10.55, 1H), 7.10 (d, J=7.77, 1H) , 7.14 (dd, J = 6.58, 4.86, 1H), 7.54 (dt, J = 7.65, 1.75, 1H), 8.45 (d J = 4.15, 1H).

Example 3 Synthesis of β-elemene-6-[two (2-pyridylmethyl)-amino]-hexylamine

0.9156g (3.8mmol) chlorinated β-elemene mixture was dissolved in 15.0mL acetonitrile, then 1.1441g (3.8mmol) of compound c was added, 0.5g NaOH was added, the temperature was controlled at 63°C, and the reaction was carried out under constant temperature reflux for 8h, extracted with chloroform, water Wash twice with brine, dry, remove solvent, use dichloromethane:methanol=20:1 (v/v) developer, flash column elution gives 1.53g brown product, beta-elemene-6-[ Bis(2-pyridylmethyl)-amino]-hexylamine (compound 3), yield 80%.

The identification results are as follows: ¹ H NMR (CDCl ₃ , TMS, 500MHz), δ: 0.97(s, 3H), 1.19-1.32(m, 4H), 1.35-1.60(m, 8H), 1.62-1.75(m, 2H), 1.69(s, 3H), 1.96-2.12(m, 2H), 2.55(t, J=6.88Hz, 2H), 2.89(t, J=7.78 Hz, 2H), 3.61(s, 2H), 3.84(s, 4H), 4.55(s, 1H), 4.80(s, 1H), 4.87-4.94(m, 2H), 5.15(s, 1H), 5.17(s, 1H), 5.79(dd, ¹ J =17.71 Hz, ² J=10.54 Hz, 1H), 7.17(t, J=5.6Hz, 2H), 7.50(d, J=7.79Hz, 2H), 7.67(dt, ¹ J=7.67Hz, ² J= 1.64Hz, 2H), 8.53 (d, J=4.19Hz, 2H); IR(KBr) v: 2928, 2362, 1388cm ^-1 ; ESI-HRMS calcd for C ₃₃ H ₄₉ N ₄ 501.3957, found 501.3922.

Example 4 Synthesis of N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemeneamine

0.2501g (1.05mmol) chlorinated β-elemene mixture was dissolved in 15.0mL acetonitrile, then 0.3310g (1.0mmol) compound d was added, 0.5g NaOH was added, the temperature was controlled at 63°C, the constant temperature reflux reaction was carried out for 8h, chloroform was extracted, and water Wash twice with brine, dry, remove solvent, use dichloromethane:methanol=20:1 (v/v) developer, flash column elution gives 0.4251g brown product, namely compound 4-N-2-{2 -[2-(Bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemeneamine, the yield was 80%.

The identification results are as follows: ¹ H NMR (CDCl ₃ , TMS, 500MHz), δ: 0.97(s, 3H), 1.35-1.63(m, 6H), 1.67(s, 3H), 1.97-2.16(m, 2H), 2.89(s, 2H), 3.22(t, J=4.32Hz, 2H), 3.46-3.51(m, 2H), 3.57-3.65(m, 4H), 3.69(s, 2H), 3.81-3.95(m, 6H), 4.54(s, 1H), 4.80(s, 1H), 4.86-4.94(m, 2H), 5.18(s, 1H), 5.32(s, 1H), 5.78(dd, ¹ J=17.85Hz, ² J=6.85Hz, 1H), 7.17(t, J=6.65Hz, 2H), 7.38(d, J=7.45Hz, 2H), 7.66(t, J=7.64Hz, 2H), 8.55(d, J =3.97Hz, 2H); IR(KBr) v: 765, 1591, 1639, 2928, 3371 cm ^-1 ; ESI-HRMS calcd for C ₃₃ H ₄₉ N ₄ O ₂ 533.3856, found 533.3845.

Example 5 Synthesis of β-elemene dipyridylamine Re(CO) ₃ complexes

1. Synthesis of rhenium tricarbonyl ([N(Et) ₄ ] ₂ [Re(CO) ₃ Br ₃ ], tetraethyl ammonium bromide tricarbonyl rhenium complex) (see the above literature)

Under the condition of nitrogen protection, weigh 0.2527g (0.6mmol) tetraethylamine bromide (Swiss Fluka company product) and 0.2076g (0.5mmol) bromopentacarbonyl rhenium (Swiss Fluka company product), add solvent diethylamine Glycol dimethyl ether 50.0mL was first slowly heated to 80°C under the condition of magnetic stirring, and the solid was completely dissolved after 10 minutes of reaction, and then heated to 120°C for 8 hours of reaction. At this time, a light yellow solid was formed in the solution. Filter while hot, wash with cold diglyme and anhydrous diethyl ether several times, and dry in a desiccator. After drying, the yellow powder was washed with absolute ethanol, filtered, and vacuum-dried to obtain the product, and 0.3727 g of a light yellow solid product was obtained, with a yield of 95%.

2. Coordination of β-elemene dipyridylmethylamine with rhenium tricarbonyl

Weigh 100 mg (0.13 mmol) of tricarbonyl rhenium and 54.1 mg (0.13 mmol) of the β-elemene dipyridylmethylamine prepared in Example 1, add 5 mL of methanol, stir at room temperature for 40 min, a small amount of powder is not dissolved, Heated at 60°C for one hour, the solution was transparent and dark brown, then the solvent was evaporated, and the solution was quickly placed on a silica gel column (silica gel: 200-300 mesh, product of China Pharmaceutical (Group) Shanghai Chemical Reagent Co., Ltd., eluent: dichloro Methane:methanol=10:1) to obtain the product as a dark brown solid 73mg, that is, β-elemene dipyridylamine Re(CO) ₃ complex (compound 5), yield 82%.

The HPLC retention time of the non-radioactive elemene dipyridylmethylamine Re complex was measured with a UV monitor to be 20.750 min, as shown in Figure 1 .

The identification data are as follows:

IR V _max : 3080 (C=CH), 2027 (C=O), 1910 (C=O);

¹ H-NMR (CDCl ₃ , TMS, 500MHz), δ: 1.03 (3H, s, CH ₃ ), 1.51-1.58 (3H, m), 1.68 (3H, s, CH ₃ ), 1.72-1.82 (2H, m), 1.97-2.02(1H, m), 2.17-2.19(1H, m), 2.30-2.39(1H, m), 4.36-4.49(4H, m), 4.61(1H, s), 4.85(1H, s), 4.91-4.99(2H, m), 5.53(1H, s), 5.62(1H, s), 5.77-5.89(1H, q), 6.05(2H, q, J=18.53Hz), 7.18(2H , t, J = 6.39Hz), 7.80 (2H, t, J = 7.63Hz), 8.0 (2H, d, J = 7.64Hz), 8.63 (2H, d, J = 5.34Hz);

¹³ C-NMR (CDCl ₃ , TMS, 125MHz), δ: 17.23, 25.64, 28.00, 34.65, 40.44, 40.82, 44.52, 52.92, 67.78, 67.92, 74.48, 110.86, 113.00, 121.66, 125.85, 1140.396, 2 , 148.02, 150.49, 151.07, 161.84, 161.95, 196.22, 196.86;

Elemental analysis results: calculated value [C ₃₀ H ₃₅ N ₃ O ₃ ReBr], C47.93%; H4.69%; N5.58%; measured value, C47.54%; H4.42%; N5.26% .

Example 6 Synthesis of β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine tricarbonyl rhenium complex

The synthesis of rhenium tricarbonyl is the same as in Example 5. Weigh 0.3852g (5.0mmol) rhenium tricarbonyl and 0.2513g (5.0mmol) of compound 3 obtained in Example 3, add 5.0mL of methanol, stir at room temperature for 40.0min, and the solution is transparent as Dark brown, then distill off the solvent, use dichloromethane:methanol=10:1 (v/v) developing solvent, quickly elute the crude product on the silica gel column, and then use dichloromethane:n-hexane=1:2 (v/v) The mixed solution was recrystallized to obtain 0.331g of a white solid, that is, β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine tricarbonyl rhenium complex (compound 6) , the yield was 78%.

The HPLC retention time of the non-radioactive compound 6 measured by an ultraviolet monitor was 21.175 min, as shown in FIG. 3 .

The identification data are as follows: m.p.161.7～162.1℃.

¹ H NMR (CDCl ₃ , TMS, 500MHz), δ: 0.99(s, 3H), 1.23-1.33(m, 2H), 1.38-1.46(m, 2H), 1.47-1.59(m, 3H), 1.62- 1.83(m, 4H), 1.70(s, 3H), 2.10-2.21(m, 4H), 2.27-2.33(m, 1H), 3.05(t, J=7.16Hz, 2H), 3.70(q, J= 14.00Hz, 2H), 3.81(t, J=8.08Hz, 2H), 4.57(s, 1H), 4.73(d, J=16.75Hz, 2H), 4.81(s, 1H), 4.87-4.89(m, 1H), 4.91(s, 1H), 5.26(s, 1H), 5.45(s, 1H), 5.59(d, J=17.02Hz, 2H), 5.82(dd, J=17.49, 10.85Hz, 1H), 7.23 (t, J = 6.71 Hz, 2H), 7.84 (dt, J = 7.74 Hz, 0.38, 2H), 7.93 (d, J = 7.81 Hz, 2H), 8.67 (d, J = 5.4 Hz, 2H).

IR (KBr) v: 3472, 2926, 2027, 1912 ^{cm -1} .

Anal. Calcd for [C ₃₆ H ₄₈ N ₄ O ₃ Re]Br.CH ₂ Cl ₂ : C47.49, H5.39, N5.99; found C47.29, H5.64, N6.26.

Example 7 N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemene

Synthesis of Amine Tricarbonyl Rhenium Complexes

The synthesis of rhenium tricarbonyl is the same as in Example 5, and 0.1193g (5.0mmol) rhenium tricarbonyl and 0.2660g (5.0mmol) N-2-{2-[2-(bis-2-pyridine) prepared in Example 4 are weighed -Methylamine)-Ethoxy]-Ethoxy}-Ethyl-N'-3-Elemeneamine (Compound 4), add 5.0mL of methanol, stir at room temperature for 40.0min, the solution is transparent and dark brown, and then evaporated Solvent, with dichloromethane: methanol = 10: 1 developer, quickly eluted on the silica gel column to give 0.3627g of crude product, to get N-2-{2-[2-(bis-2-pyridine-methylamine )-ethoxy]-ethoxy}-ethyl-N'-3-elemenylamine tricarbonyl rhenium complex (compound 7).

The HPLC retention time of the non-radioactive compound 7 measured by an ultraviolet monitor was 21.187 min, as shown in FIG. 5 .

The identification data are as follows: ¹ H NMR (CDCl ₃ , TMS, 500MHz), δ: 1.00 (s, 3 H), 1.38-1.47 (m, 2H), 1.48-1.61 (m, 2H), 1.74 (s, 3H) , 1.65-1.93(m, 10H), 2.08-2.31(m, 6H), 3.01-3.09(m, 2H), 3.62-3.74(m, 2H), 3.76-3.85(m, 2H), 4.58(s, 1H), 4.73(d, J=16.80Hz, 2H), 4.81(s, 1H), 4.87(s, 1H), 4.91(d, J=8.8Hz, 1H), 5.27(s, 1H), 5.48( s, 1H), 5.64(d, J=16.80Hz, 2H), 5.82(dd, J=17.49Hz, 10.85, 1H), 7.23(t, J=6.80Hz, 2H), 7.84(t, J=7.60 Hz, 2H), 7.95 (d, J = 7.60 Hz, 2H), 8.68 (d, J = 6.80 Hz, 2H).

IR(KBr)v: 1914, 2028, 2928, 3437 ^{cm -1} .

Example 8 Synthesis of ¹⁸⁸ Re-labeled β-elemene dipyridylamine

1) Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ synthesis (Schibli R, Schwarzbach R, Alberto R, et al. Steps Toward High Specific Activity Labeling of Biomolecules for Therapeutic Application: Preparation of Precursor[ ¹⁸⁸ Re (H ₂ O) ₃ (CO) ₃ ] ⁺ and Synthesis of Tailor-Made Bifunctional Ligand Systems. Bioconjugate Chem, 2002, 13(4): 750-756.):

a. Take vials A and B, weigh 5.6mg BH ₃ ·NH ₃ into the vials, cap and seal the remaining BH ₃ ·NH ₃ with argon gas for protection;

b. Inject CO into vials A and B filled with BH ₃ ·NH ₃ for 20 minutes at a medium flow rate;

c. Take the vial C, add 15 μL concentrated phosphoric acid (mass concentration 85%) to it, and then take 2.5mL ¹⁸⁸ ReO ₄ ^-solution ( ¹⁸⁸ W- ¹⁸⁸ Re medical nucleus provided by the Drug Release Center of Shanghai Institute of Applied Physics) with a syringe. Primer generator rinse) into bottle B, mix well;

d. Take out 1mL of the mixed solution from bottle C and put them in bottles A and B. At this time, gas is generated. Insert the syringe to keep the balance, and then put it on a water bath at 75°C to react for 15 minutes. After taking it out, cool it in an ice-water bath to obtain fac -[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ ;

e. First wet the SEP-Pak column (plus QMA WAT020545) with distilled water, and then use the column to separate the above-mentioned cooled product to remove ¹⁸⁸ ReO ₄ ^- .

2) Steps of radiolabeling the β-elemene dipyridylmethylamine ligand with Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ^] +:

Take 0.4 mL of Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ solution, add 10 ^-3 mol/L ethanol solution of β-elemene dipyridylmethylamine, compound of formula 1, 1.2 mL, seal with Seal the membrane well, place it on an incubator, control the temperature at 75°C, rotate at 1300 rpm for 50 min, and then use high-performance liquid chromatography (HPLC) [HPLC system: Merck-Hitachi L-7000 system (connected with a UV detector) and EG&GBerthold LB 508 radioactive detector), the chromatographic column is C-18ec reversed-phase column, Φ3mm*250mm, 5μm; HPLC elution gradient (solution A: anhydrous methanol; solution B: 0.05M TEAP triethylamine phosphate buffer solution , pH=2.25) gradient: 0-3min: 100%B, 0%A; 3-6min, 75%B, 25%A; 6-9min: 66%B, 34%A; 9-20min: 0%B , 100% A; 20-22min: 0% B, 100% A; 22-28min: 66% B, 34% A; 28-34min: 75% B, 25% A; 34-40min: 100% B, 0 %A; flow rate: 1mL/min] for analysis and separation, the radiochemical purity after separation was 95%. The retention time of the radioactive dipyridylamine Re complex in HPLC under the same conditions as measured by a radioactive monitor is 21.23 min, as shown in FIG. 2 .

Example 9 Synthesis of ¹⁸⁸ Re-labeled β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine

The synthesis of Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ is the same as in Example 8, except that compound 3 is replaced by compound 1 in the labeling process, and the rest is the same as in Example 8.

The radioactive β-elemene-6-[bis(2-pyridylmethyl)-amino]-hexylamine Re complex was measured with a radioactive monitor and had a retention time of 21.21min under the same conditions in HPLC. After separation, the radiochemical Pure up to 95%, as shown in Figure 4.

Example 10 ¹⁸⁸ Re-labeled N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemenamine synthesis

The synthesis of Fac-[ ¹⁸⁸ Re(CO) ₃ (H ₂ O) ₃ ] ⁺ is the same as in Example 8, except that compound 4 is replaced by compound 1 in the labeling process, and the rest is the same as in Example 8.

Radioactive N-2-{2-[2-(bis-2-pyridine-methylamine)-ethoxy]-ethoxy}-ethyl-N'-β-elemenamine Re The retention time of the complex in HPLC under the same conditions is 21.190 min, and the radiochemical purity reaches 95% after separation, as shown in FIG. 6 .

The raw materials or reagents not specified above are all conventional commercially available products.

Test Example Anticancer Activity Test

Study on anticancer activity against Hela and LLC cancer cell lines

Through the preliminary observation of anticancer activity in vitro, compared with β-elemene, the newly synthesized β-elemene tricarbonyl rhenium complex has significantly lower IC ₅₀ values on mouse LLC and human Hela cell lines (tested by t between the two groups). Test p<0.01), that is, the anticancer activity was significantly enhanced. The results are shown in Table 1.

Table 1 Inhibitory effect of β-elemene tricarbonyl rhenium complex on Hela and LLC cancer cell lines

compound IC₅₀(μM) Hela LLC β-Elemene Example 5 Example 6 Example 7 236.2±3.2 11.2±1.4^** 10.9±1.2^** 10.3±1.0^** 346.1±41.5 7.8±1.5^** 5.0±1.9^** 4.8±1.2^**

Note: ^** p<0.01

Claims (10)

1. beta-elemene derivative, its structural formula is as shown in the formula shown in the I:

Wherein, A represents

M=0-2 wherein, n=1-6.

2. beta-elemene derivative as claimed in claim 1 is characterized in that described m=0, n=2 or 6; Perhaps m=2, n=2.

3. the synthetic method of a beta-elemene derivative as claimed in claim 1 or 2, it comprises chloro β Elemenum and formula V compound is reacted in acetonitrile solvent,

Wherein, R ¹For

Wherein m, n are with definition in claim 1 or 2.

4. synthetic method as claimed in claim 3, the mol ratio that it is characterized in that this chloro β Elemenum and formula V compound is 1-3: 1, also adding is 1-12.5 with the mol ratio of formula V compound: 1 basic catalyst, temperature of reaction are 60-70 ℃, and the reaction times is 8～21 hours.

5. beta-elemene Re title complex, its structural formula is as shown in the formula shown in the II:

Wherein, definition in A such as the claim 1.

6. the synthetic method of a beta-elemene Re title complex as claimed in claim 5, it comprises formula I compound and three rhenium carbonyls in methanol solution, in room temperature to 60 ℃ reaction;

Wherein, definition in A such as the claim 1.

7. the application of beta-elemene Re title complex as claimed in claim 5 in the preparation antitumor drug.

8. one kind ¹⁸⁸The beta-elemene derivative of Re mark, shown in the following formula III of its structural formula:

Wherein, definition in A such as the claim 1.

One kind as claimed in claim 8 ¹⁸⁸The synthetic method of the beta-elemene derivative of Re mark, it comprises employing Fac-[ ¹⁸⁸Re (CO) ₃(H ₂O) ₃] ⁺Formula I compound is carried out radio-labeling;

Wherein, definition in A such as the claim 1.

10. synthetic method as claimed in claim 9 is characterized in that described radio-labeling comprises Fac-[ ¹⁸⁸Re (CO) ₃(H ₂O) ₃] ⁺Solution adds in the ethanolic soln of formula I compound, adopts couveuse, 75 ℃ of temperature controls, and 1300 rev/mins of rotating speeds in 50 minutes reaction times, adopt HPLC to separate then.

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