10-hydroxybenzoquinoline platinum complexes, process for their synthesis and use thereof

Abstract

The present application provides 10-hydroxybenzoquinoline platinum complex and its synthesis method and application, belongs to the medical technology field, 10-hydroxybenzoquinoline platinum complex chemical formula is: C 15 H 15 ClNO2PtS.The platinum complex L-Pt shows significant inhibitory effect on MDA-MB-231 cancer cells, and the IC 50 Value is 0.092±0.01 μM, far lower than the clinical antitumor drug cisplatin (8.86±0.79 μM);Compared with the synthesis raw material ligand L and cis-Pt (DMSO) 2Cl2 of platinum complex L-Pt, MDA-MB-231 cancer cells have almost no inhibitory effect, platinum complex L-Pt has high tumor selectivity on MDA-MB-231 cancer cells, 10-hydroxybenzoquinoline platinum complex L-Pt shows excellent antitumor activity, shows its potential value as drug development, and is expected to become a variety of antitumor drug preparation candidate.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to 10-hydroxybenzoquinoline platinum complex and its synthesis method and application. BACKGROUND

[0002] Although platinum drugs have achieved great success in the clinical application of treating cancer, platinum drugs have the disadvantages of serious toxic side effects and easy drug resistance. Quinoline alkaloids are a class of alkaloids with quinoline ring as the basic mother nucleus, such as antimalarial drug quinine and anticancer drug camptothecin. Hydroxyquinoline derivatives are intermediates of various drugs and have wide pharmacological activities, such as anticancer, anti-AIDS, antifungal, antischistosome, antioxidant and neuroprotective activities. Therefore, it is necessary to design 10-hydroxybenzoquinoline platinum complex and its synthesis method and application. SUMMARY

[0003] The purpose of the present application is to provide 10-hydroxybenzoquinoline platinum complex and its synthesis method and application, which solves the technical problem that there is no effective drug for treating breast cancer.

[0004] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows:

[0005] 10-hydroxybenzoquinoline platinum complex, chemical formula is: C 15 H 15 ClNO2PtS, chemical structure formula is:

[0006]

[0007] The synthesis method of 10-hydroxybenzoquinoline platinum complex is as follows: 0.3g of K2PtCl4 is weighed and dissolved in 10mL of H2O, continuously stirred at room temperature, and dimethyl sulfoxide (DMSO) is added dropwise after 10min until the solution is yellow, heated to 65℃, stirred for 30min, cooled to room temperature, transferred to a 4℃ refrigerator, and filtered the next day, and washed with acetone three times.

[0008] The synthesis of platinum complex L-Pt is as follows: 0.39g of ligand L is dissolved in 10mL of ethanol and 5mL of acetone, and 0.85g of platinum intermediate [Pt(DMSO)2Cl2] (2mmol) is dissolved in 5mL of ethanol and 5mL of acetone. The [Pt(DMSO)2Cl2] solution is slowly added to the L solution at room temperature, and the reaction temperature is controlled at 63-67℃, at which time the solution is light yellow. The solution is refluxed for 6h under water bath stirring, and cooled overnight. The next day, the filtrate is transferred to a beaker, sealed with plastic wrap, and then slowly volatilized at room temperature for 7d to obtain yellow needle-shaped crystals, which are 10-hydroxybenzoquinoline platinum complex.

[0009] 10-Hydroxybenzoquinoline platinum complex is applied to preparation of anti-breast cancer drugs.

[0010] Further, 10-Hydroxybenzoquinoline platinum complex is applied to preparation of anti-breast cancer drugs, and the proliferation inhibition activity experiment of human breast cancer cell MDA-MB-231 is carried out.

[0011] Further, the specific process of the proliferation inhibition activity experiment is as follows:

[0012] (2) Preparation of L-Pt solution: a proper amount of L-Pt complex is weighed, dissolved in DMSO to prepare a 10mmol·L -1 solution;

[0013] (2) MDA-MB-231 tumor cells in logarithmic growth phase are taken, washed twice with 5mL PBS buffer, digested with 1mL trypsin, terminated digestion with 5mL medium and gently blown off the adherent cells, and then the cells are evenly mixed and inoculated in a 96-well culture plate;

[0014] (3) The growth of cells is observed under a microscope, when the confluence of cells reaches more than 80%, 20μL of prepared L-Pt is taken by a pipette and added to each well, and 5 parallel holes are set for the drug administration group and the blank group, and the experiment is repeated three times;

[0015] (4) The culture plate is placed in a 37℃, 5% CO2 incubator, and cultured for another 48h, then the plate is taken out, 20μL of 5mg / mL [4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide reagent is added, the plate is evenly shaken for four times, and then the plate is placed in the incubator for another 4h;

[0016] (5) The plate is taken out, the culture solution is poured out, 200μL of DMSO solution is added to each well, and the plate is placed on a horizontal shaker and shaken gently for 15min;

[0017] (6) The absorbance value is measured at 570nm by using an enzyme-labeled instrument.

[0018] The present application has the following beneficial effects due to the adoption of the above technical solutions:

[0019] The platinum complex L-Pt shows significant inhibition effect on MDA-MB-231 cancer cells, and the IC 50The value is 0.092±0.01 μM, which is much lower than that of the clinical anti-tumor drug cisplatin (8.86±0.79 μM); in comparison, the synthesis raw material ligand L of the platinum complex L-Pt and cis-Pt(DMSO)2Cl2 have little inhibitory effect on MDA-MB-231 cancer cells. The platinum complex L-Pt has high tumor selectivity for MDA-MB-231 cancer cells. The 10-hydroxybenzoquinoline platinum complex L-Pt exhibits excellent anti-tumor activity, showing its potential value as a drug development and being expected to become a candidate for the preparation of various anti-tumor drugs. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a crystal structure diagram of the 10-hydroxybenzoquinoline platinum complex of the present application. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical scheme and advantages of the present application more clear and explicit, the present application is further described in detail below with reference to the drawings and preferred embodiments. However, it should be noted that many details in the description are only to make the reader have a thorough understanding of one or more aspects of the present application, and these aspects can be realized without these specific details.

[0022] 1. Synthesis of 10-hydroxybenzoquinoline platinum complex

[0023] Synthesis of platinum intermediate [Pt(DMSO)2Cl2]: weigh 0.3 g of K2PtCl4 and dissolve in 10 mL of H2O, continuously stir at room temperature, add dimethyl sulfoxide (DMSO) drop by drop after 10 min until the solution is yellow, warm to 65℃, stir for 30 min, cool to room temperature, transfer to a 4℃ refrigerator, and filter the next day, wash with acetone three times.

[0024] Synthesis of platinum complex L-Pt: weigh 0.39 g of ligand L (2 mmol) and dissolve in 10 mL of ethanol and 5 mL of acetone, weigh 0.85 g of platinum intermediate [Pt(DMSO)2Cl2] (2 mmol) and dissolve in 5 mL of ethanol and 5 mL of acetone, slowly add the [Pt(DMSO)2Cl2] solution to the L solution at room temperature, control the reaction temperature at about 65℃, at this time the solution is light yellow, reflux the reaction under water bath stirring for 6 h, cool overnight, filter the next day, transfer the filtrate to a small beaker, seal with plastic wrap, and then slowly evaporate at room temperature for 7 d to obtain yellow needle-shaped crystals, the yield is about 55%. Select suitable single crystals for X-ray diffraction analysis to determine the crystal structure, as shown in Figure 1 .

[0025] 2. Inhibition activity experiment of platinum complex L-Pt on the proliferation of human breast cancer cells MDA-MB-231

[0026] (1) Preparation of L-Pt solution: Weigh an appropriate amount of L-Pt complex and dissolve it in DMSO to prepare 10 mmol·L -1 of solution.

[0027] (2) MDA-MB-231 tumor cells in the logarithmic growth phase were taken, washed twice with 5 mL of PBS buffer, digested with 1 mL of trypsin, and terminated with 5 mL of culture medium. The adherent cells were gently blown off, and the cells were mixed and inoculated into a 96-well culture plate;

[0028] (3) Observe the cell growth under a microscope. When the cell confluence reaches more than 80%, pipette 20 μL of the prepared L-Pt into each well. Set up 5 replicate wells in parallel for the drug-treated group and the blank group, and repeat the experiment three times.

[0029] (4) Place the culture plate in a 37°C, 5% CO2 incubator for 48 h. Remove the plate, add 20 μL of 5 mg / mL MTT ([4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) reagent, pat the plate evenly, and return the plate to the incubator for 4 h.

[0030] (5) Remove the plate, discard the culture medium, add 200 μL of DMSO solution to each well, place the plate on a horizontal shaker, and gently shake for 15 minutes;

[0031] (6) Use an enzyme-labeled instrument to measure the absorbance at 570 nm.

[0032] 3.1 Structural characterization of platinum complexes

[0033] Analysis of X-ray single crystal diffraction data shows that L-Pt belongs to the monoclinic system with a space group of P21 / c. 10-Hydroxybenzoquinoline (L) reacts with [Pt(DMSO)2Cl2] to synthesize [Pt(DMSO)LCl]. Pt(II) coordinates with the hydroxyl O atom in 10-hydroxybenzo[h]quinoline and the N atom of the quinoline heterocycle, while retaining the original DMSO and Cl atoms to form a tetracoordinate complex. The crystal structure of L-Pt is shown in Figure 1. Figure 1 The crystallographic data and structural modification parameters are shown in Table 1, and some bond lengths and bond angles are shown in Table 2.

[0034] Table 1 L-Pt crystallographic data and structural correction parameters

[0035]

[0036]

[0037] Table 2 Partial bond lengths of L-Pt and bond angle (°)

[0038]

[0039] 3.2 L-Pt inhibits tumor cell growth experiment results

[0040] Table 3. IC50 values of compounds on MDA-MB-231 cell line (μM)

[0041]

[0042]

[0043] According to the IC 50 screening results, the platinum compound L-Pt exhibits significant inhibitory effect on MDA-MB-231 cancer cells, with an IC 50 value of 0.092±0.01 μM, which is much lower than that of the clinical anti-tumor drug cisplatin (8.86±0.79 μM); in comparison, the synthesis raw material ligand L and cis-Pt(DMSO)2Cl2 of the platinum compound L-Pt have little inhibitory effect on MDA-MB-231 cancer cells. This result shows that the platinum compound L-Pt has high tumor selectivity on MDA-MB-231 cancer cells. In summary, the 10-hydroxybenzoquinoline platinum complex L-Pt exhibits excellent anti-tumor activity, showing its potential value as a drug development, and is expected to become a candidate for the preparation of various anti-tumor drugs.

[0044] The details of the present application are known in the art.

[0045] The above description is only preferred embodiments of the present application, and it should be pointed out that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. 10-Hydroxybenzoquinoline platinum complex, characterized by: The chemical formula is: C15H15ClNO2PtS, and the chemical structure is:

2. The method for synthesizing a 10-hydroxybenzoquinoline platinum complex according to claim 1, wherein: Synthesis of platinum intermediate [Pt(DMSO)2Cl2]: Weigh 0.3g K2PtCl4 and dissolve it in 10mL H2O. Stir continuously at room temperature. After 10 minutes, add dimethyl sulfoxide (DMSO) dropwise until the solution turns yellow. Heat to 65°C, stir for 30 minutes, cool to room temperature, and refrigerate at 4°C. Filter the solution the next day and wash three times with acetone. Synthesis of platinum complex L-Pt: Weigh 0.39 g of ligand L and dissolve it in 10 mL of ethanol and 5 mL of acetone, and 0.85 g of platinum intermediate [Pt(DMSO)2Cl2] (2 mmol) and dissolve it in 5 mL of ethanol and 5 mL of acetone. Slowly add [Pt(DMSO)2Cl2] solution dropwise to the L solution at room temperature. The reaction temperature is controlled at 63-67°C. At this time, the solution is light yellow. Reflux the solution under stirring in a water bath for 6 hours, cool overnight, and filter the solution the next day. Transfer the filtrate to a beaker, seal it with plastic wrap, and then slowly evaporate it at room temperature for 7 days to precipitate yellow needle-like crystals, which are 10-hydroxybenzoquinoline platinum complex.

3. Application of 10-hydroxybenzoquinoline platinum complex, characterized by: 10-Hydroxybenzoquinoline platinum complexes are used to prepare anti-breast cancer drugs.

4. The use of the 10-hydroxybenzoquinoline platinum complex according to claim 3, characterized in that: 10-Hydroxybenzoquinoline platinum complex is used in the preparation of anti-breast cancer drugs, and the proliferation inhibitory activity of human breast cancer cells MDA-MB-231 is first tested.

5. The use of the 10-hydroxybenzoquinoline platinum complex according to claim 4, characterized in that: The specific process of the proliferation inhibition activity experiment is as follows: (1) Preparation of L-Pt solution: Weigh an appropriate amount of L-Pt complex and dissolve it in DMSO to prepare 10 mmol·L -1 of solution; (2) MDA-MB-231 tumor cells in the logarithmic growth phase were taken, washed twice with 5 mL of PBS buffer, digested with 1 mL of trypsin, and terminated with 5 mL of culture medium. The adherent cells were gently blown off, and the cells were mixed and inoculated into a 96-well culture plate; (3) Observe the cell growth under a microscope. When the cell confluence reaches more than 80%, pipette 20 μL of the prepared L-Pt into each well. Set up 5 replicate wells in parallel for the drug-treated group and the blank group, and repeat the experiment three times. (4) Place the culture plate in a 37°C, 5% CO2 incubator for 48 h. Remove the plate, add 20 μL of 5 mg / mL [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reagent, pat the plate evenly, and return the plate to the incubator for 4 h. (5) Remove the plate, discard the culture medium, add 200 μL of DMSO solution to each well, place the plate on a horizontal shaker, and gently shake for 15 minutes; (6) Use an enzyme-labeled instrument to measure the absorbance at 570 nm.

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