Multi-target cis-platinum complex as well as synthesis method and anticancer activity application thereof

Abstract

The invention discloses a multi-target cis-platinum (CDDP)-platinum (IV) complex as well as a preparation method and application thereof, and belongs to the field of medicinal chemistry. The complex has two different bioactive ligands in the axial position, cis-platinum is used as a raw material, cis-platinum is oxidized into oxyplatinum in hydrogen peroxide, and then the oxyplatinum, axial ligand NHS ester and anhydride are subjected to continuous two-time carboxylation to obtain a three-effect platinum (IV) anticancer prodrug candidate. The first carboxylation is carried out by adopting 1 / 1 NHS ester and oxyplatinum in DMSO, and the second carboxylation is carried out by adopting 20 / 1 anhydride and a monosubstituted complex in DMF; the reaction raw materials are easy to obtain, the synthesis steps are simple, and the reaction conditions are mild. The multi-target cis-platinum complex shows extremely strong physiological activity to cancer cells such as A549, HepG2 and MCF-7, has the median inhibitory concentration of less than 1 mu M, and can be further used in potential anti-cancer drugs.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and specifically relates to a synthesis method of a multi-target Pt (IV) complex and its anti-tumor application. Background Art

[0002] Cancer remains the leading cause of death in humans. The main means of cancer treatment are radiotherapy and chemotherapy. Nearly half of patients use platinum chemotherapy to treat cancer. Cisplatin, as the first generation of platinum chemotherapy drugs, has a simple structure and a clear mechanism of action. After entering the body, cisplatin enters cells from the bloodstream passively or actively through copper transporters. Cisplatin is first hydrolyzed in the cell, and then binds to the N7 position of the nuclear DNA purine base through the Pt(II) center, distorting the DNA double-stranded structure, thereby inducing cell apoptosis. Although cancer cells are more sensitive to cisplatin than healthy tissue cells, only a small part of the drug enters the cancer cell during the application of platinum drugs, and a considerable part of the drug will interact with DNA and other nucleophiles in normal cells, resulting in off-target. This off-target platinization leads to dose-limiting toxic side effects in the human body, and drug resistance seriously limits the widespread application of platinum drugs.

[0003] In the past few decades, it has been demonstrated that inflammation predisposes to cancer and promotes all stages of tumorigenesis. Cyclooxygenase (COX) is a key enzyme in the biosynthesis of arachidonic acid-derived prostaglandins. COX-1 is expressed in almost all tissues and is responsible for regulating the basal level of prostaglandins in normal physiological processes. COX-2 can be induced in inflammatory and tumorigenic tissues and can mediate overexpression of prostaglandin E2 (PGE2). PGE2 plays an important role in tumorigenesis, invasion, metastasis and drug resistance. Widely used nonsteroidal anti-inflammatory drugs such as diclofenac and naproxen can inhibit COX activity, thereby preventing the formation of prostaglandins and inflammatory effects.

[0004] Histones are proteins that control the structure of chromatin and nucleosomes. Histone deacetylase (HDAC) deacetylates histones, making DNA more tightly wrapped around histones, making it difficult for these DNAs to be contacted by gene transcription factors. This inhibits the expression of proteins related to cell cycle arrest, tumor immunity, and apoptosis of damaged cells, all of which promote the development of cancer. Histone deacetylase inhibitors (HDACi) can selectively restore the expression of these cancer suppressors and other anti-cancer genes by controlling the tightness of DNA wrapped around histones. HDACi can also indirectly inhibit the expression of angiogenic factors and help block the blood supply to tumors. 4-phenylbutyric acid (PhB) is also a known HDACi that binds to coenzyme A (CoA) through thiol adducts and inhibits histone deacetylase (HDAC), thereby affecting cell metabolism. Its synergistic effect with DNA-binding drugs has attracted attention as an anti-cancer compound.

[0005] Cinnamic acid (Cin) is a naturally occurring phytochemical with a variety of pharmacological activities, including antimicrobial, anti-inflammatory, antioxidant, and antitumor activities, with no significant effects on normal cells. In malignant tumor cell lines, Cin induces tumor cell apoptosis and differentiation by inhibiting the activity of matrix metalloproteinases (MMPs), inhibiting invasive growth and metastasis, while inducing cell cycle arrest and cytoskeletal disruption. Cin can also affect the differentiation of sphere-derived cancer stem cells (CSCs), making them more sensitive to cisplatin through apoptosis, and reducing the toxic effects of cisplatin on normal cells.

[0006] To overcome these limitations and improve the anti-cancer effect, clinicians use platinum drugs in combination with other drugs for chemotherapy. The main advantage of combined chemotherapy is that several antiproliferative drugs with different mechanisms of action and different cellular targets attack the tumor, thereby increasing the chance of killing cancer cells and overcoming resistance to a single drug. Currently, researchers are conducting (pre)clinical studies on the combined use of several non-steroidal anti-inflammatory drugs such as aspirin and ibuprofen with chemotherapy drugs to understand their anti-tumor potential. However, each drug has different pharmacokinetic properties, and combined use can complicate the overall treatment results.

[0007] Platinum (IV) prodrugs with low spin octahedral structures have the potential to overcome the side effects and drug resistance development of FDA-approved platinum (II) anticancer drugs cisplatin, carboplatin, and oxaliplatin. They are kinetically more inert than their platinum (II) precursors and can be administered orally, a feature that can reduce the difficulty of drug administration and improve the quality of life of cancer patients. As prodrugs, Pt (IV) complexes are activated by reduction inside cancer cells, releasing two axial ligands as well as square planar Pt (II) drugs. Due to the reduction of Pt (IV) prodrugs inside cells, axial ligands with specific therapeutic properties can be released, making the actual therapeutically active Pt (II) drug and its bioactive ligand a perfect combination for multi-action therapy. Based on this combination strategy, previous researchers designed a series of dual-target multi-specific Pt (IV) complexes with non-steroidal anti-inflammatory drugs as axial ligands, such as naproxen, indomethacin, ibuprofen, etc., combined with Pt (IV), and explored their anticancer activities. Summary of the invention

[0008] In order to overcome the above technical defects, the purpose of the present invention is to provide the synthesis, characterization and chemical and biological properties of multi-target Pt (IV) complexes, which are prepared by connecting diclofenac or naproxen at one axial position and using hydroxyl coupling Cin or PhB at the second axial position to prepare triple-acting prodrugs. A series of biological studies were conducted to study its antiproliferative effects on a variety of different cancer cells (Hela, SW480 and A549, etc.) and the accumulation of Pt in MCF-7 cells. The results show that the three-target active group Pt (IV) complex based on cisplatin has greatly improved the enhanced anti-tumor activity compared to cisplatin, showing better anti-tumor potential.

[0009] The multi-target Pt (IV) complex of the present invention has the following structural formula:

[0010]

[0011] The present invention also provides a method for synthesizing the above-mentioned multi-target Pt (IV) complex. The technical solution is to first convert cisplatin into a single-coordinated dual-target tetravalent platinum prodrug, and then convert it into a multi-target tetravalent platinum prodrug compound.

[0012] The synthesis method comprises the following steps: using cisplatin, phenylbutyric acid, naproxen and diclofenac as raw materials, and obtaining multi-target cisplatin complexes 1-3 through oxidation, esterification, carboxylation and other steps;

[0013] Specifically, the steps include:

[0014] Step A, cisplatin a is oxidized with hydrogen peroxide under light-proof conditions to generate oxyplatinum b; diclofenac c / phenylbutyric acid f and N-hydroxysuccinimide are condensed in the presence of dicyclohexylcarbodiimide to generate intermediates d / j respectively; phenylbutyric acid f / cinnamic acid h / naproxen l are condensed in the presence of dicyclohexylcarbodiimide to generate anhydrides g / i / m respectively;

[0015] Step B, heating oxygen platinum b and intermediate d / j to react to generate intermediate e / k;

[0016] Step C: The intermediate e / k reacts with the anhydride g / i / m by heating at elevated temperature to generate multi-target Pt(IV) complexes 1-3.

[0017] The synthetic route is shown below:

[0018]

[0019] Among them: the axial ligands of cisplatin, diclofenac and naproxen, are both cyclooxygenase (COX) inhibitors; the axial ligand phenylbutyric acid is a histone deacetylase (HDAC) inhibitor; and cinnamic acid is a matrix metalloproteinase (MMP) inhibitor.

[0020] Furthermore, in step A of the above technical solution, the concentration of hydrogen peroxide is 20-30%, and the reaction temperature is 60-80°C.

[0021] Furthermore, in step A of the above technical solution, the molar ratio of diclofenac / phenylbutyric acid to N-hydroxysuccinimide (NHS) is 1:1, and the molar ratio of diclofenac / phenylbutyric acid to dicyclohexylcarbodiimide (DCC) is 1:1.

[0022] Furthermore, in step A of the above technical solution, the molar ratio of phenylbutyric acid / cinnamic acid / naproxen to dicyclohexylcarbodiimide (DCC) is 1:0.5.

[0023] Furthermore, in step B of the above technical scheme, the molar ratio of the intermediate d / j to the oxygen platinum b is 1:1; the reaction is carried out in DMSO solvent, and the reaction temperature is selected from 60-70°C.

[0024] Furthermore, in step C of the above technical solution, the molar ratio of the intermediate e / k to the anhydride g / i / m is 10:1; the reaction is carried out in DMF solvent, and the reaction temperature is selected to be 75-90°C.

[0025] The present invention also provides the application of the multi-target cisplatin complex in preparing anti-tumor drugs.

[0026] Furthermore, in the above technical solution, the anti-tumor is anti-Hela, MCF-7, HepG2, HCT116, SW480, A549 tumor cells; the complex has the best activity against A549 cells.

[0027] Advantageous Effects of the Invention

[0028] 1. The present invention uses cisplatin and phenylbutyric acid, cinnamic acid, diclofenac and naproxen as raw materials, splices together different compounds targeting tumor targets, and innovatively constructs three new multi-target cisplatin complexes.

[0029] 2. The multi-target cisplatin complex of the present invention was tested for its activity on Hela, MCF-7, HepG2, HCT116, SW480 and A549, wherein the cisplatin complex showed extremely strong cell activity against drug-resistant A549 cancer cells.

[0030] 3. The multi-target cisplatin complex of the present invention was tested for the total cell platinum accumulation in MCF-7, wherein: the accumulation of the multi-target cisplatin complex in MCF-7 cancer cells was five times that of cisplatin, showing extremely strong cancer cell killing performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1It is the accumulated amount of platinum of the multi-target cisplatin complex in MCF-7 cells in the example. Specific embodiments

[0032] The present invention will be further described below by specific examples. These embodiments should be understood to be only used to illustrate the present invention and not to limit the scope of protection of the present invention. After reading the content of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

[0033] This reaction was modified using cisplatin as a template substrate, and different tumor target inhibitors were used to splice cisplatin to find the best reaction conditions, resulting in three extremely active multi-target cisplatin complex prodrugs.

[0034] Example 1

[0035] Taking compound 1 as an example, the experimental operation steps are:

[0036] 30wt% H 2 O 2 The (70mL) solution was added dropwise to a round-bottom flask containing cisplatin a (1.00g, 3.33mmol). The mixture was heated to 70°C and stirred vigorously for 4 hours under light-proof conditions. The suspension became a bright yellow transparent solution. The reaction was terminated and placed at 4°C overnight. Bright yellow crystals precipitated the next day. The crystals were filtered and recovered, washed with cold water, ethanol and ether, and vacuum dried to obtain yellow crystals of 0.89g of oxyplatinum b.

[0037] Weigh diclofenac c (2.96 g, 10 mmol) and N-hydroxysuccinimide (1.15 g, 10 mmol) and dissolve in 50 mL CHCl. 3 DCC (2.06 g, 10 mmol) was added and stirred at room temperature for 7 h. After the reaction was completed, the dicyclohexylurea precipitate was removed by filtration, and the filtrate was concentrated in vacuo to obtain a white solid, which was purified by column chromatography (methanol / dichloromethane = 1:20) to obtain 1.86 g NHS-diclofenac ester d.

[0038] Weigh phenylbutyric acid f (3.28 g, 20 mmol) and DCC (1.15 g, 10 mmol) and dissolve them in 50 mL CH 2 Cl 2 The mixture was stirred at room temperature for 7 hours. After the reaction, the dicyclohexylurea precipitate was removed by filtration. The filtrate was concentrated in vacuo to obtain a waxy white solid. The residue was dissolved in ethyl acetate and allowed to stand overnight at -20°C. The insoluble residue was removed by filtration the next day, washed with petroleum ether and dried; purified by column chromatography (methanol / dichloromethane = 1:20) to obtain 2.14 g of phenylbutyric anhydride g.

[0039] Weigh NHS-diclofenac d (196.6 mg, 0.5 mmol) and platinum oxide b (166 mg, 0.5 mmol) and dissolve them in 10 mL DMSO. The mixture is stirred at 65°C in the dark for 48 h. At this time, the reaction liquid is a clear and transparent light yellow solution. After the reaction is completed, filter and remove a large amount of DMSO by vacuum rotary evaporation to obtain a yellow viscous solid. Freeze-dry to remove the remaining DMSO, and then dissolve the residual solid and phenylbutyric anhydride g (1.55 g, 5 mmol) in 10 mL DMF. The mixture is stirred at 80°C in the dark for 48 h. At this time, the reaction liquid is a clear and transparent light yellow solution. Then evaporate the solvent, freeze-dry to remove DMF, and purify by column chromatography (methanol / dichloromethane = 1:20) to obtain 65 mg of product 1.

[0040] Multi-target cisplatin complex 1 (65 mg); yield 17%; 1 H NMR (400 MHz, DMSO-d 6 )δ7.51(d,J=8.1Hz,2H),7.27(m,3H),7.18(m,5H),7.04(td,J=7.7,1.6Hz,1H),6.82(td,J=7.4,1.2Hz,1H),6.56(s, 6H), 6.25 (dd, J=8.1, 1.2Hz, 1H), 3.77 (s, 2H), 2.60 (t, J=7.6Hz, 2H), 2.23 (t, J=7.4Hz, 2H), 1.76 (hept, J=6.5Hz, 2H). 13 C NMR(101MHz,DMSO-d6)δ181.26,179.30,157.12,143.59,142.46,137.87,131.4 4,130.75,130.13,129.51,128.96,128.78,128.70,127.63,126.29,126.16,12 5.79,125.68,120.83,116.13,79.65,55.38,47.98,36.27,35.62,34.90,33.82,31.75,31.25,29.49,29.30,29.04,28.00,27.02,25.79,24.93,22.56,14.42. 195 Pt NMR(86MHz,DMSO-d6)δ1228.40; HRMS(ESI)m / z:[M+Na] + Calcd for C 24 H 27 Cl 4 N 3 O4 PtNa 780.0273; Found 780.0287.

[0041] Compounds 2 and 3 were obtained by a similar method to compound 1, and the product characteristics are as follows: Multi-target cisplatin complex 2 (105 mg); yield 28%; 1 H NMR (400 MHz, DMSO-d 6 )δ7.63(m,2H),7.52(d,J=8.1Hz,2H),7.40(m,4H),7.27(s,1H),7.18(ddd,J=8.0,5.0,3.4Hz,2H),7.04(t d,J=7.6,1.6Hz,1H),6.83(m,1H),6.65(s,6H),6.55(d,J=15.9Hz,1H),6.26(d,J=8.0Hz,1H),3.79(s,2H); 13 CNMR (101MHz, DMSO-d 6 )δ179.31,174.41,162.80,143.61,141.64,137.88,134.96,131.47,130.84,130.20,129.53,129. 42,128.67,128.31,127.66,125.86,125.65,121.80,120.83,116.13,55.40,40.90,36.27,31.25; 195 Pt NMR (86 MHz, DMSO-d 6 )δ1219.55; HRMS(ESI)m / z:[M+Na] + Calcd for C 23 H 23 Cl 4 N 3 O 4 PtNa 763.9960; Found 763.9958.

[0042] Multi-target cisplatin complex 3 (70 mg); yield 20%; 1 H NMR (400 MHz, DMSO-d 6)δ7.74(m,3H),7.47(dd,J=8.5,1.9Hz,1H),7.26(m,3H),7.21(d,J=1.6Hz,1H),7.19(s,1H),7.14(m,2H),6.53(s,6H ),3.86(s,3H),3.82(m,1H),2.58(m,2H),2.23(t,J=7.4Hz,2H),1.75(p,J=7.6Hz,2H),1.42(dd,J=15.3,7.1Hz,3H). 13 C NMR (101 MHz, DMSO-d 6 )δ182.50,181.03,157.58,157.40,142.47,137.86,133.71,133.51,129.56,128.95,128.83,128.69,127.47,127.3 2,126.82,126.15,126.04,125.96,119.16,118.84,106.13,55.60,46.92,36.25,35.45,34.92,27.97,20.26,18.90. 195 PtNMR (86 MHz, DMSO-d 6 )δ1218.53; HRMS(ESI)m / z:[M+Na] + Calcd for C 24 H 30 Cl 2 N 2 O 5 PtNa 715.1060; Found715.1100.

[0043] Example 2

[0044] This example is the test results of the in vitro anti-proliferation experiment of the compounds described in Example 1 against various cancer cells, namely the half-inhibitory concentration and the accumulation of platinum in cells. Unless otherwise specified, the materials and consumables listed in this example can be obtained from commercial channels, and the cells are from the cell and microbial resource library of CTCC or other relevant institutions. The experimental method of this example is a standard molecular biology, cell biology or virology operation procedure, which can be easily understood and operated by researchers in the field. The specific steps are as follows:

[0045] 1. Cells:

[0046] The cell lines used were: human cervical cancer cell line Hela, human liver cancer cell line HepG-2, human breast cancer cell line MCF-7, human colon cancer cell line SW480, human colon cancer cell line HCT116, human lung cancer cell line A549, and human lung cancer cisplatin-resistant cell line A549 (A549 / CDDP). Among them: Hela, HepG-2, MCF-7, A549, HCT116 cell lines were cultured with DMEM complete medium, SW480 cell line was cultured with L-15 complete medium, and human lung cancer cisplatin-resistant A549 cell line was cultured with A549 / CDDP-specific F-12K complete medium. The above cell lines were all cultured at 37°C, 5% CO 2 The cells were cultured in a humidified cell incubator.

[0047] 2. Main reagents:

[0048] DMEM medium;

[0049] Fetal bovine serum (FBS);

[0050] L-15 medium

[0051] 3. In vitro anti-proliferation experimental steps of tumor cells:

[0052] Step 1: Observe the state of cells after subculturing. When they are in the logarithmic growth phase, digest and centrifuge them, then blow and mix them evenly. Count them with a cell counter and dilute the cell suspension to 3-8×10 3 Cells / 100 μL were seeded in a 96-well plate, with 100 μL in each well.

[0053] Step 2: Cultivate cells: Place the 96-well plate with cells in a 37°C, 5% CO 2 Culture the cells in a saturated humidity cell incubator for 12-24 hours, and treat with drugs after the cells are fully attached.

[0054] Step 3: Drug administration: In a clean bench, in the dark, use blank culture medium to dilute the mother solution of the compound to be tested to a 10-fold drug solution with the corresponding concentration gradient; carefully remove the original culture medium and add 180 μL of fresh complete culture medium; add 20 μL of drug solution to the corresponding wells of the 96-well plate at each well, that is, dilute 10 times to achieve the corresponding drug concentration gradient, and the DMSO concentration is less than 0.1%. Set up 5 replicates for each concentration, and set up a blank control group. Shake gently to mix, mark, and place at 37°C, 5% CO 2 The cells were cultured in a humidified cell incubator for 48 h.

[0055] Step 4 Color development: After 48 h, remove the culture medium from the 96-well plate, add 100 μL of fresh complete culture medium and 10 μL of CCK8 (5 mg / mL) solution, shake gently in parallel to mix, and place at 37°C, 5% CO 2 Continue culturing in a saturated humidity cell incubator for 1-5 hours.

[0056] Step 5: Determination: Finally, the absorbance value (OD value) at λ = 450 nm was measured using an ELISA reader; IC was estimated from the concentration-response curve using GraphPad Prizm 8.01 software. 50 Values ​​were analyzed by nonlinear regression using curve fitting.

[0057] 4. Experimental steps for determination of Pt content in whole cells

[0058] Step 1: Digest the MCF-7 cells in the logarithmic growth phase, centrifuge them, collect them in a centrifuge tube, count them, and adjust the cell suspension concentration to 1×10 6 The cells were inoculated into a 6-well plate, 2 mL per well, i.e. 1×10 6 cells / well, incubate at 37°C, 5% CO 2 The cells were cultured overnight in a cell incubator under saturated humidity.

[0059] Step 2: Drug administration: After the cells are completely attached, the drug is administered. The mother solution of the compound to be measured is diluted to 1 μM with the culture medium and the cells are administered. A cisplatin control group and a drug group are set up, with 3 wells in each group. The cells are placed at 37°C and 5% CO. 2 The cells were cultured overnight in a cell incubator under saturated humidity.

[0060] Step 3 Collection: After removing the culture medium, wash the 6-well plate 3 times with cold PBS, add 500 μL of 0.25% trypsin to digest until the cells become round and the edges become clear, remove the trypsin, add 2 mL of complete culture medium and gently blow until the cells fall off, transfer the cells to a centrifuge tube, centrifuge at 2000 rpm for 5 minutes, discard the supernatant and wash with PBS 2 times, add 1 mL of PBS and gently blow evenly, count, centrifuge at 12000 rpm for 1 minute, collect the cells and transfer to ep tube for freeze-drying. Then seal and store in a -80℃ refrigerator for later use.

[0061] Step 4 determination: After taking out the freeze-dried cells, use a pipette to accurately measure 200 μL of concentrated nitric acid for overnight digestion, add 1 mL of ultrapure water for dilution, and use ICP-MS to determine the Pt content.

[0062] 5. Test results:

[0063] The results of the in vitro antiproliferative activity of the three studied prodrugs against tumor cells at gradient concentrations are as follows;

[0064] <![CDATA[IC 50 / μM]]> CDDP 1 FI 2 FI 3 FI Hela 4.20±0.06 0.26±0.01 16.2 0.13±0.01 33.1 0.13±0.004 31.3 MCF-7 18.46±1.25 0.31±0.02 58.7 0.27±0.01 69.2 0.19±0.02 96.5 HepG-2 12.81±0.57 0.26±0.01 49.7 0.32±0.02 39.5 0.25±0.01 51.2 SW480 29.21±2.36 0.21±0.01 137.6 0.18±0.01 162.8 0.15±0.01 201.3 HCT116 15.46±0.22 0.01±0.001 1260.0 0.01±0.001 1387.8 0.03±0.001 477.3 A549 4.33±0.18 0.10±0.002 42.8 0.11±0.003 39.3 0.11±0.005 40.8 A549 / CDDP 28.16±0.50 0.40±0.02 71.1 0.28±0.004 101.9 0.35±0.02 81.0

[0065] Fold increase (FI) = IC 50 (Cisplatin) / IC 50 (Prodrug compound).

[0066] The results of the antiproliferative activities of the three studied prodrugs against cisplatin-resistant strains are as follows:

[0067] <![CDATA[IC 50 / μM]]> A549 A549 / CDDP RF CDDP 4.33±0.18 28.16±0.50 6.5 1 0.10±0.002 0.40±0.02 3.9 2 0.11±0.003 0.28±0.004 2.5 3 0.11±0.005 0.35±0.02 3.3

[0068] Resistance Factor(RF)=IC 50 (Human lung cancer cisplatin-resistant A549 cell line) / IC 50 (Human lung cancer A549 cell line).

[0069] The same concentration of three prodrugs (compound concentration: 1 μM) was used to treat MCF-7 cells (1×10 6 ) after the whole cell Pt content ( Figure 1 ).

[0070] in conclusion:

[0071] 1. The most obvious conclusion from the cytotoxicity data is that complexes 1-3 all exhibit nanomolar antiproliferative activity against most tested cell lines, which is dozens of orders of magnitude higher than cisplatin. Especially for the HCT116 cell line, the half-inhibitory concentrations of complexes 1 and 2 are both around 10nM (FI>1000), which is more than 1000 times higher than cisplatin, showing extremely high anti-tumor activity.

[0072] 2. Regarding the selectivity of the drug for drug-resistant cell lines, the anti-proliferative activity of the three studied prodrugs against cisplatin-resistant strains was improved to varying degrees, that is, the resistance factors were all less than the resistance factor of cisplatin for drug-resistant strains, 6.509 (RF<6.509). In addition, multi-target complexes 1-3 have higher selectivity for drug-resistant cell lines, which indicates that multi-target complexes have great potential for the treatment of drug-resistant cells.

[0073] 3. ICP-MS was used to detect the intracellular Pt content of MCF-7 cells treated with 1 μM compound for 24 h. Figure 1 As shown, the Pt contents in whole cells treated with compounds 1-3 were 52.8 ng Pt / 10 6 cells、57.6ng Pt / 10 6 cells、58.3ng Pt / 10 6cells, which is about 5 times that of the group treated with the same dose of cisplatin. Since cellular uptake or efflux is the first step in its anti-tumor mechanism, the accumulation of small molecule platinum chemotherapy drugs in tumor cells plays a key role in their anti-proliferative activity.

[0074] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. Three multi-target Pt(IV) complexes, characterized in that: The structural formula is as follows: Among them: the axial ligands of cisplatin, diclofenac and naproxen, are both cyclooxygenase (COX) inhibitors; the axial ligand phenylbutyric acid is a histone deacetylase (HDAC) inhibitor; and the axial ligand cinnamic acid is a matrix metalloproteinase (MMP) inhibitor.

2. The method for synthesizing a multi-target Pt(IV) complex according to claim 1, characterized in that: The method comprises the following steps: using cisplatin, naproxen / diclofenac, phenylbutyric acid / cinnamic acid as raw materials, and obtaining multi-target cisplatin complexes 1-3 through oxidation, esterification, carboxylation and other steps.

3. The method for synthesizing a multi-target cisplatin complex according to claim 2, characterized in that: The specific steps include: Step A, cisplatin a is oxidized with hydrogen peroxide under light-proof conditions to generate oxyplatinum b; diclofenac c / phenylbutyric acid f and N-hydroxysuccinimide are condensed in the presence of dicyclohexylcarbodiimide to generate intermediates d / j respectively; phenylbutyric acid f / cinnamic acid h / naproxen l are condensed in the presence of dicyclohexylcarbodiimide to generate anhydrides g / i / m respectively; Step B, heating oxygen platinum b and intermediate d / j to react to generate intermediate e / k; Step C: The intermediate e / k reacts with the anhydride g / i / m by heating at elevated temperature to generate multi-target Pt(IV) complexes 1-3.

4. The method for synthesizing the multi-target cisplatin complex according to claim 3, characterized in that: In step A, the concentration of hydrogen peroxide is 20-30%, and the reaction temperature is 60-80°C.

5. The method for synthesizing the multi-target cisplatin complex according to claim 3, characterized in that: In step A, the molar ratio of diclofenac / phenylbutyric acid to N-hydroxysuccinimide is 1:1, and the molar ratio of diclofenac / phenylbutyric acid to dicyclohexylcarbodiimide is 1:

1.

6. The method for synthesizing the multi-target cisplatin complex according to claim 3, characterized in that: In step A, the molar ratio of phenylbutyric acid / cinnamic acid / naproxen to dicyclohexylcarbodiimide is 1:0.

5.

7. The method for synthesizing the multi-target cisplatin complex according to claim 3, characterized in that: In step B, the molar ratio of the intermediate d / j to the oxygen platinum b is 1:1; the reaction is carried out in DMSO solvent, and the reaction temperature is selected from 60-70°C.

8. The method for synthesizing the multi-target cisplatin complex according to claim 3, characterized in that: In step C, the molar ratio of the intermediate e / k to the anhydride g / i / m is 10:1; the reaction is carried out in DMF solvent, and the reaction temperature is selected to be 75-90°C.

9. Use of the multi-target cisplatin complex as claimed in claim 1 in the preparation of anti-tumor drugs.

10. The use of the multi-target cisplatin complex according to claim 1 in the preparation of an anti-cisplatin resistant tumor drug, characterized in that: The anti-tumor agents are against Hela, MCF-7, HepG2, HCT116, SW480, and A549 tumor cells.

Images