CN116375769A - Double-target Pt (IV) anticancer prodrug, and preparation method and application thereof - Google Patents

Abstract

The invention discloses a double-target Pt (IV) anticancer prodrug, a preparation method and application thereof, and the chemical name is fac- [ Pt (1R, 2R-cyclohexanediamine) (hydroxy) (3-bromopyruvate) ₃ ]I.e. fac- [ Pt (1R, 2R-DACH) (OH) (3-bromoxyruvate) ₃ ]The oxaliplatin prodrug contains 3-bromopyruvate (glycolytic inhibitor) ligands in the molecule, can inhibit the replication of tumor cell DNA and glycolytic pathway simultaneously, and has two action targets. Its synthesis is based on cis- [ Pt (1R, 2R-DACH) I ₂ ]Or cis- [ Pt (1R, 2R-DACH) Cl ₂ ]As a starting material, after hydrolysis of silver sulfate, it was reacted with an alkali metal hydroxide to give cis- [ Pt (1R, 2R-DACH) (OH) ₂ ]The solution is then subjected to an axial oxidation with hydrogen peroxide to form the intermediate cis- [ Pt (1R, 2R-DACH) (OH) ₄ ]·H ₂ And finally, carrying out neutralization reaction with 3 times of 3-bromopyruvate to obtain the target complex. The invention is characterized in thatThe Pt (IV) anticancer prodrug has good water solubility and stability of aqueous solution, strong antitumor effect, low toxicity and low cross drug resistance with oxaliplatin, and is used for chemotherapy of malignant tumor.

Description

A dual-target Pt(IV) anticancer prodrug and its preparation method and application

technical field

The invention relates to a dual-target Pt(IV) anticancer prodrug and its preparation method and application. The Pt(IV) anticancer prodrug is a Pt(IV) complex with a chemical formula of fac-[Pt( 1R,2R-cyclohexanediamine)(hydroxyl)(3-bromopyruvate) ₃ ], namely fac-[Pt(1R,2R-DACH)(OH)(3-bromopyruvate) ₃ ], oxaliplatin A prodrug containing 3 3-bromopyruvic acid (glycolysis inhibitor, 3-bromopyruvic acid) ligands. The Pt(IV) anticancer prodrug of the present invention can inhibit the DNA replication and glycolysis pathway of tumor cells at the same time, has two action targets, has good water solubility and high antitumor activity, can be used for chemotherapy of malignant tumors, and belongs to biological pharmaceutical field.

Background technique

Malignant tumors are the second leading cause of death after cardiovascular and cerebrovascular diseases. Although the targeted therapy and immunotherapy that appeared in recent years have greatly improved the prognosis of tumor clinical treatment, surgery, radiotherapy and chemotherapy are still indispensable basic therapies for the clinical treatment of malignant tumors, especially chemotherapy, which targets tumor cells. The characteristics of heterogeneity and rapid proliferation show unique advantages, and combined application with targeted therapy and immunotherapy can make up for the deficiencies of targeted and immunotherapy in this regard. The basis of chemotherapy is chemotherapy drugs. Platinum anticancer drugs represented by cisplatin (DDP), carboplatin (CBP) and oxaliplatin (OXP) are a very important class of chemotherapy drugs, and their targets are Cancer cell DNA has strong anticancer activity and unique mechanism of action, and is compatible with most natural drugs (such as paclitaxel), targeted drugs (such as gefitinib, trastuzumab), immunotherapeutic drugs (such as PD-1/PD -L1 immunosuppressant pembrolizumab/atezolizumab) has a synergistic anticancer effect and is widely used in the clinical treatment of common multiple malignant tumors. According to the latest statistics, more than 50% of chemotherapy regimens are based on platinum anticancer drugs or have platinum anticancer drugs in combination. However, there are still two major drawbacks in the clinical application of platinum-based anticancer drugs: toxic side effects and drug resistance. Therefore, the development of new platinum-based drugs with high anticancer activity, low toxicity, and low cross-resistance to existing drugs still has important clinical value.

The chemical structures of cisplatin, carboplatin and oxaliplatin are:

Platinum drugs currently used clinically, including cisplatin, carboplatin, and oxaliplatin, are all Pt(II) complexes. Pt(II) belongs to the d ⁸ valence electron configuration and uses d ² sp ³ hybrid , forming a four-coordinate planar configuration, and the kinetics of the coordination substitution reaction is relatively active. Before reaching the DNA target of cancer cells, it is easy to have unwanted chemical reactions with biomolecules in blood or intercellular fluid, resulting in toxic and side effects. At the same time, some drugs were also degraded. The Pt(II) complex forms a hexacoordinated octahedral Pt(IV) complex after axial oxidation, and the general structural formula is cis, trans, cis-[Pt(IV)A ₂ Y ₂ X ₂ ] (where , A is the pharmacophore, Y is the axial ligand, X is the leaving group), the reaction kinetics is inert, the reaction rate with biomacromolecules is significantly reduced, and it can maintain relative stability in the in vivo environment , with low toxicity. Moreover, tumor tissue is a reducing microenvironment, and the concentration of glutathione and vitamin C is significantly higher than that of normal cells, which can reduce Pt(IV) to the corresponding Pt(II) anticancer drug cis-[Pt(II)A _{2X2 ]} . Therefore, Pt(IV) complexes can be regarded as prodrugs of Pt(II) anticancer drugs, providing an effective way for the tumor-targeted delivery of Pt(II) anticancer drugs. In addition, choosing an appropriate axial ligand Y can also adjust the water-solubility and fat-solubility of Pt(IV) complexes, increase the target to improve anti-cancer activity, etc. Therefore, Pt(IV) prodrugs are one of the hot research directions of platinum-based anticancer drugs in recent years.

The energy supply of human cells mainly comes from glucose, and there are two pathways: mitochondrial oxidative phosphorylation and glycolysis. Normal cells mainly rely on mitochondrial oxidative phosphorylation for energy; unlike normal cells, tumor cells lack metabolic flexibility and mainly supply energy through the glycolytic pathway, which is an adaptive response to intermittent hypoxia, even in normal This energy supply mode is also maintained under oxygen concentration, which is the so-called Warburg effect, but the energy produced by the glycolysis process is limited. Therefore, in order to maintain rapid proliferation and other activities, tumor cells will accelerate the body’s intake and metabolism of sugar. This is obvious. The characteristics make the glycolytic pathway an important target for antitumor action. 3-Bromopyruvate is a small organic carboxylic acid molecule that can inhibit the glycolysis pathway by inhibiting the activity of hexokinase II (the first rate-limiting enzyme of glycolysis), resulting in the inability of tumor cells to absorb energy and die, while It has no effect on surrounding normal tissues and cells; moreover, 3-bromopyruvate can also overcome the multidrug resistance of tumors and enhance the activity of anticancer drugs such as platinum anticancer drugs. However, 3-bromopyruvate is relatively unstable, and it will degrade soon after entering the human body, and has no obvious antitumor effect. Therefore, the stability of 3-bromopyruvate has become the main limiting factor affecting its antitumor effect.

Contents of the invention

The technical problem solved by the present invention is to introduce 3-bromopyruvate into the Pt(IV) prodrug molecule through the coordination substitution reaction with the axial and radial hydroxyl groups of Pt(IV), in order to increase the concentration of the Pt(IV) prodrug target, while improving the stability of 3-bromopyruvate.

The present invention uses cis-[PtA ₂ I ₂ ] or cis-[PtA ₂ Cl ₂ ] as the starting material, and carries out hydrolysis reaction with equimolar AgSO ₄ to generate cis-[PtA ₂ (OH ₂ ) ₂ ](SO ₄ ), and then react with an equimolar alkali metal hydroxide to convert into a cis-[PtA ₂ (OH) ₂ ] solution. After axial oxidation with hydrogen oxide, the intermediate cis-[PtA ₂ (OH) ₄ ] is obtained. H ₂ O, the synthetic route of the target dual-target Pt(Ⅳ) anticancer prodrug can be expressed as:

The above synthetic route is suitable for 2A being 2NH ₃ , ethylenediamine, 1,2-propylenediamine, 1,3-propylenediamine, 1R, 2R-DACH and other ammonia/amine ligands and their mixed amines. Finally, the intermediate The body is neutralized with 3-bromopyruvate in order to form a Pt(IV) complex of the following chemical formula:

(1) fac-[Pt(Ⅳ)A ₂ (OH)(3-bromopyruvate) ₃ ];

(2) cis-[Pt(IV)A ₂ (3-bromopyruvate) ₄ ].

The inventor successfully synthesized a Pt(IV) complex fac-[Pt(1R,2R-DACH)(OH)(3-bromopyruvate) ₃ ] containing 3 molecules of 3-bromopyruvate through multiple studies and experiments (code name: BrPt-4), is the prodrug of oxaliplatin, the yield is about 82%, and its chemical structural formula is:

A method for preparing a dual-target Pt (IV) anticancer prodrug, comprising the following steps:

Step (1): At 45-60°C, cis-[Pt(1R,2R-DACH)I ₂ ] or cis-[Pt(1R,2R-DACH)Cl ₂ ] and equimolar Ag ₂ SO ₄ in Stir and react in the aqueous solution for 6 hours in the dark, and filter to remove silver iodide or silver chloride precipitate to obtain cis-[Pt(1R,2R-DACH)(OH ₂ ) ₂ ](SO ₄ ) solution;

Step (2): Stir and react cis-[Pt(1R,2R-DACH)(OH ₂ ) ₂ ](SO ₄ ) and equimolar alkali metal hydroxide at 15-30°C for 1-4 hours, filter Remove alkali metal sulfate (MSO ₄ ) precipitate to obtain cis-[Pt(1R,2R-DACH)(OH) ₂ ] solution;

Step (3): Add excess hydrogen peroxide dropwise to the cis-[Pt(1R,2R-DACH)(OH) ₂ ] solution at 30-50°C, stir and react for 4 hours, and spin the obtained solution under reduced pressure Evaporate to the minimum volume, add a large amount of ice acetone for reverse analysis, filter, dry, and then recrystallize through a water-acetone mixed solvent to obtain the intermediate cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O;

Step (4): cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O and 3-fold molar amount of 3-bromopyruvate were stirred and reacted in aqueous solution at 30～40°C for 48～ After 72 hours, the obtained solution was lyophilized, filtered, washed with ether, dried, and recrystallized from methanol to obtain the target complex fac-[Pt(1R,2R-DACH)(OH)(3-bromopyruvate) ₃ ].

Further, the alkali metal hydroxide described in step (2) is any one of strontium hydroxide, calcium hydroxide and barium hydroxide.

Further, the molar ratio of cis-[Pt(1R,2R-DACH)(OH) ₂ ]:H ₂ O ₂ in step (3) is 1:10-25.

Its reaction process is as follows:

The inventor found in the synthesis test that in the aqueous solution system used in the synthesis, even if the amount of 3-bromopyruvate is increased to 6 times the calculated amount, four 3-bromopyruvate ligands cannot be introduced simultaneously, and its structure is:

The isolated product was still BrPt-4 containing three 3-bromopyruvate groups, which may be due to the fact that when the three hydroxyl groups in cis-[Pt(IV)A ₂ (OH) ₄ ]·H ₂ O were After 3-bromopyruvate is substituted, the pK _a value of 3-bromopyruvate is not enough to replace the fourth hydroxyl in the molecule.

BrPt-4 is a prodrug of oxaliplatin with a water solubility of 12.1mg/mL. At the same time, observing its ¹ H-NMR changes over time in D ₂ O shows that there is no significant change in ¹ H-NMR at room temperature for 72 hours. Therefore, BrPt-4 has good aqueous solution stability and meets the requirements of platinum complexes. Conditions required for the preparation of medicines.

Using the MTT method, the Pt(Ⅳ) complex BrPt-4 of the present invention was measured against human non-small cell lung cancer cell line (A549), human ovarian cancer cell line (SKOV3), human gastric cancer cell line (MKN-28), human colon The proliferation of cancer cell line (HCT116) and human liver cancer cell line (HepG2) both have high inhibitory activity, indicating that BrPt-4 has high anti-cancer cell proliferation activity in vitro. Moreover, most importantly, the inhibitory activity of BrPt-4 on the proliferation of oxaliplatin-resistant human non-small cell lung cancer cell line (A549/OXP) was significantly higher than that of oxaliplatin, suggesting that BrPt-4 has considerable resistance Cancer cell drug resistance.

In the in vivo model of transplanted tumor S180 in mice, after intraperitoneal injection of BrPt-4, the tumor inhibition rate was significantly higher than that of oxaliplatin. From the perspective of changes in routine and myeloid hyperplasia, except for the slightly larger weight difference of mice before and after administration, other toxicity indicators of BrPt-4 were significantly lower than those of oxaliplatin, indicating that BrPt-4 has high anti-tumor activity and comparative Little toxicity.

According to the results reported in literature at home and abroad, the antitumor activity of Pt(IV) complexes is generally inferior to that of the corresponding Pt(II) drugs, which is probably due to the insufficient reduction of Pt(IV) to Pt(II). The antitumor activity of BrPt-4 is higher than that of oxaliplatin, which may partly come from the antitumor effect of 3-bromopyruvate (3-BrPA), that is, BrPt-4 has two targets, and its antitumor mechanism can be Expressed as:

In addition to inhibiting the DNA replication of tumor cells, the Pt(IV) complex BrPt-4 of the present invention can also obtain additional anti-tumor effects by inhibiting the glycolysis pathway of tumor cells.

In summary, the Pt(Ⅳ) complex BrPt-4 of the present invention has good water solubility, high stability, high antitumor activity and low toxicity, and can be used as a prodrug of oxaliplatin. In the treatment of malignant tumors (such as lung cancer, ovarian cancer, gastric cancer, colon cancer and liver cancer, etc.).

Description of drawings

Figure 1: The chemical structural formula of the Pt(IV) complex BrPt-4 of the present invention.

Figure 2: Schematic diagram of the effect of intraperitoneal injection (ip) administration of the test compound on the bone marrow of tumor-bearing mice.

Detailed ways

Example 1: Synthesis of intermediate cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O

Add cis-[Pt(1R,2R-DACH)I ₂ ](6.26g, 11.12mmol) or cis-[Pt(1R,2R-DACH)Cl ₂ ](4.23g, 11.12mmol) into 60mL of water and stir to form To a uniform paste, add an aqueous solution (110mL) of silver sulfate (3.40g, 10.90mmol), stir at 45-60°C in the dark for 6 hours, check the reaction to the end, filter, and wash with water three times to obtain a light yellow clear Add equimolar alkali metal hydroxide (M(OH) ₂ ) to the filtrate, stir in the dark at 15-30°C for 1-4 hours, filter to obtain a colorless and clear filtrate, and place the filtrate in a water bath at 30-50°C , add 30wt% hydrogen peroxide (25.21～63.01g) dropwise, continue to stir and react for 4 hours, and the reaction solution is rotary evaporated under reduced pressure to the minimum volume to obtain light yellow oil, add a large amount of ice acetone for reverse analysis, and white to pale The yellow solid was filtered, washed twice with ice acetone, and dried in vacuum for 8 hours. The obtained crude product was recrystallized from a water-acetone mixed solvent to obtain 3.80 g cis-[Pt(1R,2R-DACH)(OH) ₄ ]· H ₂ O, yield 86.26%.

Structural characteristic parameters: <1>Elemental analysis: measured value Pt 49.12%, C 17.96%, H 5.88%, N 6.81% (calculated value Pt 49.40%, C 18.22%, H 5.06%, N 7.09%).

Example 2: Synthesis of target complex fac-[Pt(1R,2R-DACH)(OH)(3-bromopyruvate) ₃ (BrPt-4)

Dissolve cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O (1.01g, 2.56mmol) in 10mL water, add 3-bromopyruvate (1.35g, 8.08mmol) in water (5mL ), stirred and reacted at 30-40° C. for 48-72 hours. During the reaction, the solution gradually precipitated a brownish-yellow solid from a clear solution. After the reaction, the reaction solution was freeze-dried, filtered, washed with ether three times, and dried in vacuum for 8 hours. The obtained brownish-yellow solid was purified by methanol recrystallization to obtain 1.71g Product, the yield is 81.12%.

Structural characteristic parameters:

(1) Elemental analysis

Measured value Pt 23.38%, C 21.47%, H 3.08%, N 3.01% (calculated value Pt 23.66%, C21.84%, H 2.55%, N 3.40%);

(2) ¹ H NMR (500MHz, DMSO-d ₆ ) δ6.31-5.49 (m, 4H, 2NH ₂ ), 3.33 (s, H ₂ O), 2.50 (p, J=1.8Hz, DMSO), 2.35 (d, J=15.8Hz, 2H, 2CH-cyclohexyl), 1.98 (d, J=12.6Hz, 2H, CH ₂ -ax-3-BrPA), 1.89 (t, J=13.6Hz, 4H, 2CH ₂ - eq-3-BrPA), 1.51(s, 2H, CH ₂ -cyclohexyl), 1.26(d, J=31.1Hz, 4H, 2CH ₂ -cyclohexyl), 1.02(t, J=10.6Hz, 2H, CH ₂ - cyclohexyl);

(3) IR (KBr, cm ^-1 ): 3435(m), 3202(m), 1687(s), 1322(s), 573(w), 513(w), 440(w);

(4) ESI-MS: 825 [M+H] ⁺ .

Example 3: Inhibitory activity of Pt(IV) complex BrPt-4 of the present invention on tumor cell proliferation in vitro

Positive control sample oxaliplatin (batch number: L20200428) was purchased from Kunming Guiyan Pharmaceutical Co., Ltd.; tumor cell lines were purchased from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

The effects of compound BrPt-4 and oxaliplatin (OXP) on cell proliferation were detected by MTT assay. Take cells in the logarithmic growth phase, including human non-small cell lung cancer cell line (A549) and its oxaliplatin-resistant strain (A549/OXP), human ovarian cancer cell line (SKOV3), human gastric cancer cell line (MKN -28), human colon cancer cell line (HCT116), human liver cancer cell line (HepG2) and human normal liver cells (L02), were routinely digested to make a single cell suspension and counted, adjusted to a certain concentration of cell suspension, inoculated Incubate in a 96-well culture plate, 90 μL/well, in a 37°C, 5% CO ₂ saturated humidity incubator for 24 hours. After the cells adhere to the wall, add different concentrations of test drugs. All compounds were formulated with dextrose injection (5% GS). According to different cells, 5 different test concentrations were set, and 4 parallel wells were set for each concentration, 10 μL/well. After adding the drug, continue to culture in the incubator for 48 hours, add 20 μL of MTT (5mg/ml) to each well, continue to cultivate for 4 hours, and absorb the supernatant, then add 100 μL of DMSO to each well to dissolve the reduced product formazan, and use a microplate reader at 570nm , measured the OD value of each well at dual wavelengths of 630nm, and calculated the inhibition rate. According to the inhibition rate of each concentration, the half inhibitory concentration IC ₅₀ was calculated by using SPSS software. The results are shown in Table 1.

Table 1. Effects of test compounds on the proliferation of different cell lines

As shown in the results in Table 1, the inventive compound BrPt-4, except that the inhibitory activity on the proliferation of human non-small cell lung cancer cell line (A549) is slightly lower than that of oxaliplatin, has no effect on the human non-small cell lung cancer cell line (A549) resistant to oxaliplatin. Small cell lung cancer cell line (A549/OXP), human ovarian cancer cell line (SKOV3), human gastric cancer cell line (MKN-28), human colon cancer cell line (HCT116) and human liver cancer cell line (HepG2) all had Very high inhibitory activity, IC ₅₀ is less than the corresponding value of oxaliplatin. In particular, the inhibitory activity on the proliferation of the oxaliplatin-resistant lung cancer cell line A549/OXP is significantly higher than that of oxaliplatin, suggesting that the invented compound has considerable drug resistance against tumor cells.

During the test, we also set up a human normal cell line (L02) for comparison to evaluate the toxicity of the test compound to normal cells. The results showed that the inhibitory concentration IC ₅₀ of BrPt-4 to the growth of the normal cell line was greater than that of the corresponding Oxa Liplatin, showing that BrPt-4 has certain selectivity for tumor cells.

Example 4: In vivo tumor inhibitory effect and preliminary toxicity evaluation of Pt(IV) complex BrPt-4 administered by intraperitoneal injection of the present invention

Kunming (KM) mice, 22-25 grams, female, were purchased from Hunan Slack Experimental Animal Co., Ltd.; mouse sarcoma S180 tumor strain was quoted from Shanghai Institute of Materia Medica, Chinese Academy of Sciences; positive control drug oxaliplatin (batch number: L20200428 ) was purchased from Kunming Guiyan Pharmaceutical Co., Ltd. Both BrPt-4 and oxaliplatin were formulated with 5% GS to the desired concentration.

Take the S180 cells of ascites-type mice that grew well 5-8 days after inoculation, adjust the cell concentration to 1.0×10 ⁷ /mL with normal saline (NS), and inoculate them subcutaneously in the right axilla of the mice, 0.2 mL/mouse, 24 hours after inoculation Divide into 3 groups at random, adopt intraperitoneal injection administration (ip), give vehicle, oxaliplatin and BrPt-4 respectively, 1 time/day, continuous 13 days, the oxaliplatin that administration dose refers to report to mice The half effective dose (ED ₅₀ ) of S180 inhibition and the preliminary test results of this study were selected as 7.6 μmol/kg. Mice were sacrificed 24 hours after the last administration, and fasted for 12 hours before sacrifice. The tumor was taken out and weighed, and the tumor inhibition rate was calculated=(average tumor weight of the control group-average tumor weight of the treatment group/average tumor weight of the control group×100%). value. The results are shown in Table 2. At the same time, the effect of the compound on body weight, vital organs and blood indicators after administration was investigated to evaluate its preliminary toxicity.

1. Overall anti-tumor effect in vivo

The experimental results are shown in Table 2. Compared with the vehicle group, oxaliplatin and BrPt-4 had significant tumor inhibitory effects, and the inhibition rates were 67.81% and 83.14%, respectively. However, the body weight gain of the mice was also affected after administration, which was statistically significant compared with the vehicle group, suggesting a toxic reaction. Considering the tumor inhibition rate and the weight change of the mice after administration, the curative effect of BrPt-4 is better than that of oxaliplatin at an equimolar dose.

Table 2. Effect of test compound intraperitoneal injection (ip) administration on the growth of transplanted tumor S180 in mice

Note: Compared with vehicle control group: *, P<0.05; **, P<0.01; ***, P<0.001

2. The effect of drug administration on important organs and blood indicators

The thymus and spleen are important immune organs and the most common toxic target organs of cytotoxic anticancer drugs. The liver and kidney are the main organs of drug metabolism and the main toxicity targets of platinum-based drugs. The results in Table 3 showed that compared with the vehicle control group, the thymus and spleen weights of the mice in the administration group were significantly reduced, suggesting that both oxaliplatin and BrPt-4 have immunosuppressive effects, but from both In terms of the reduction degree of the compound, the immunosuppressive effect of oxaliplatin is stronger than that of BrPt-4. However, at the dose of 7.6 μmol/kg, oxaliplatin and BrPt-4 had little effect on the liver-kidney coefficient of mice.

Table 3. Effects of test compound intraperitoneal injection (ip) administration on spleen, thymus, liver and kidney weights of tumor-bearing mice

Note: Compared with vehicle group: *, P<0.05; **, P<0.01; ***, P<0.001.

Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are indicators of liver function that reflect whether or not the liver cells are damaged and how severe they are. They all exist in liver cells. When liver cell membranes are damaged or cells are necrotic, these enzymes enter the serum and increase; Serum creatinine CREA concentration can accurately reflect the damage degree of glomerular filtration function to a certain extent. When the renal function is normal, the creatinine excretion rate is constant, and when the renal parenchyma is damaged, the glomerular filtration rate will decrease. When the filtration rate drops to a certain level, the blood creatinine concentration will rise sharply; BUN is a metabolite of human protein, which is mainly filtered by the glomerulus and excreted with urine. When the kidney parenchyma is damaged, The glomerular filtration rate decreases, resulting in an increase in blood urea nitrogen concentration, so by measuring blood urea nitrogen, we can understand the glomerular filtration function. Table 4 shows the changes in liver and kidney function indexes of S180 mice bearing oxaliplatin and BrPt-4 by intraperitoneal injection (ip). Compared with the vehicle group, BrPt-4 had little effect on the liver and kidney function, but the AST in the oxaliplatin group was increased with statistical difference, suggesting that it had an effect on the liver function. Oxaliplatin has little effect on renal function, which is consistent with the reports at home and abroad.

Table 4. Effect of test compound intraperitoneal injection (ip) administration on liver and kidney function of tumor-bearing mice

Note: Compared with vehicle control group: *, P<0.05; **, P<0.01.

Bone marrow suppression is also the most common toxicity of platinum-based drugs, often dose-limiting toxicity, and is the main factor causing death in mice in acute toxicity tests. Bone marrow suppression leads to a decrease in the number of blood cells, especially platelets (PLT) and white blood cells (WBC). The results in Table 5 show that after administration of oxaliplatin and BrPt-4 to tumor-bearing mice, WBC, PLT, and RBC (red blood cells) all showed a downward trend in varying degrees, and the decline in the oxaliplatin group was particularly obvious. There was a significant difference in group comparison, indicating that oxaliplatin had more severe myelosuppression than BrPt-4.

Further mouse sternal bone marrow smear test can be seen (as shown in Figure 2), compared with the vehicle control group, the bone marrow hyperplasia of the mice in the oxaliplatin group was in an extremely reduced state, while the bone marrow in the BrPt-4 group was in an active state, Tip: The bone marrow toxicity of oxaliplatin is greater than that of BrPt-4 of the present invention.

Table 5. Effect of test compound intraperitoneal injection (ip) administration on blood routine of tumor-bearing mice

Note: Compared with the negative control group: *, P<0.05; **, P<0.01; ***, P<0.001.

Claims (10)

1. a double-target Pt (Ⅳ) anticancer prodrug, characterized in that the chemical name of the Pt (Ⅳ) anticancer prodrug is fac-[Pt (1R, 2R-cyclohexanediamine) (hydroxyl) (3-bromopyruvate) ₃ ], containing three glycolysis inhibitor 3-bromopyruvate ligands in the molecule, the chemical structural formula is:

2. a kind of preparation method of a kind of dual-target Pt (IV) anticancer prodrug according to claim 1, is characterized in that, comprises the following steps: Step (1): At 45-60°C, cis-[Pt(1R,2R-DACH)I ₂ ] or cis-[Pt(1R,2R-DACH)Cl ₂ ] and equimolar Ag ₂ SO ₄ in Stir the reaction in an aqueous solution in the dark, filter to remove silver iodide or silver chloride precipitate, and obtain cis-[Pt(1R,2R-DACH)(OH ₂ ) ₂ ](SO ₄ ) solution; Step (2): Stir and react cis-[Pt(1R,2R-DACH)(OH ₂ ) ₂ ](SO ₄ ) and equimolar alkali metal hydroxide at 15-30°C, and remove alkali metal sulfuric acid by filtration Salt (MSO ₄ ) was precipitated to obtain cis-[Pt(1R,2R-DACH)(OH) ₂ ] solution; Step (3): Add excess hydrogen peroxide dropwise to the cis-[Pt(1R,2R-DACH)(OH) ₂ ] solution at 30-50°C, stir for reaction, and rotate the obtained solution under reduced pressure, Add ice acetone for reverse analysis, filter, dry, and then recrystallize from a water-acetone mixed solvent to obtain the intermediate cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O; Step (4): cis-[Pt(1R,2R-DACH)(OH) ₄ ]·H ₂ O and 3-fold molar amount of 3-bromopyruvate were stirred and reacted in aqueous solution at 30-40°C to obtain The solution was lyophilized, filtered, washed with ether, dried, and recrystallized from methanol to obtain the target complex fac-[Pt(1R,2R-DACH)(OH)(3-bromopyruvate) ₃ ]. 3. preparation method according to claim 2 is characterized in that, the alkali metal hydroxide described in step (2) is any one in strontium hydroxide, calcium hydroxide, barium hydroxide. 4. preparation method according to claim 2, is characterized in that, described in step (3) cis-[Pt(1R,2R-DACH)(OH) ₂ ]:H _{2 O} mol ratio is 1: 10-25. 5. according to the preparation method described in any one of claim 2-4, it is characterized in that, in step (1), avoid light and stir reaction for 6 hours. 6. The preparation method according to any one of claims 2-4, characterized in that, in step (2), the reaction is stirred for 1-4 hours. 7. The preparation method according to any one of claims 2-4, characterized in that, stirring and reacting for 4 hours in step (3). 8. The preparation method according to any one of claims 2-4, characterized in that, in step (4), the reaction is stirred for 48-72 hours. 9. A use of a dual-target Pt(IV) anticancer prodrug according to claim 1 in the preparation of antitumor drugs for clinical chemotherapy of malignant tumors. 10. The use according to claim 9, characterized in that the malignant tumors include lung cancer, ovarian cancer, gastric cancer, colon cancer and liver cancer.

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