Metal antitumor drug TTP of platinum complex and its synthetic method and application

Abstract

The present application relates to the technical field of antitumor drug synthesis, and particularly relates to a metal antitumor drug TTP of platinum complex, and a synthesis method and application thereof, wherein 2-bromopyridine and 5-methyl 2-aldehyde pyridine are used as raw materials, halogen-lithium exchange is carried out by reacting with butyllithium and reacting with bromopyridine, a hydroxyl dipyrrole compound is obtained by anionic attack on pyridine formaldehyde, a carbon-based dipyrrole compound is further prepared by oxidation, bromination is carried out by NBS, condensation is carried out by triphenylphosphine, and complexation is carried out by potassium tetrachloroplatinate, and finally the final product TTP which targets mitochondria is obtained. The present application can inhibit two energy metabolism pathways of mitochondrial oxidative phosphorylation and glycolysis, reduce the overall metabolic level of tumor cells, and finally achieve the purposes of inhibiting tumor cell proliferation and overcoming cell drug resistance. The raw material is cheap and easy to obtain, the structure is novel, the yield is high, the separation and purification operation is simple, mass production is suitable, the product shows good biological activity, and detection shows that the metal antitumor drug TTP has an IC 50 . of 5.17 uM for non-small cell lung cancer A549.

Description

Technical Field

[0001] This invention relates to the field of antitumor drug synthesis technology, and in particular to a platinum complex metal antitumor drug TTP, its synthesis method and application. Background Technology

[0002] With the development of the times, the average lifespan of humans has been continuously increasing and living standards have been gradually improving. Cancer has become another major disease threatening human health, yet no drug has been discovered that can completely cure cancer. Mitochondria are tiny organelles within cells that provide almost all the energy needed for cellular function by producing ATP. Mitochondria are widely involved in cellular processes such as signal transduction, energy metabolism, autophagy, and apoptosis, and are crucial for maintaining the normal physiological functions of organisms. At the same time, their functional impairment is also closely related to the occurrence and development of various diseases such as cancer and Alzheimer's disease. Mitochondrial function requires the participation of various proteins and inorganic substances to be completed, so the unique physicochemical properties of metal complexes can be used for intervention or regulation to achieve the purpose of preventing or treating diseases. The energy metabolism process of cancer cells differs from that of normal cells. They preferentially choose aerobic glycolysis as the glucose metabolism pathway. This abnormal energy metabolism process involves numerous proteins and enzymes, thus providing potential targets for designing anticancer drugs. Platinum-based anticancer drugs have long been the main chemotherapy drugs for treating cancer. These drugs are generally believed to react with nuclear DNA and induce apoptosis by inhibiting nuclear DNA replication and gene transcription. However, they all have certain toxic side effects and drug resistance. Their clinical efficacy is severely weakened by drug resistance. The main reason is that the accumulation of drugs in cells decreases and the cell's self-repair ability increases. Therefore, there is an urgent need to develop a drug that is low in toxicity and does not easily induce drug resistance.

[0003] Cisplatin is the most widely used metallo-based anticancer drug in clinical practice; however, addressing its toxicity and resistance remains a challenge in the pharmaceutical field. Glycolysis occurs via two pathways: mitochondrial oxidative phosphorylation and glycolysis. In normal mammalian cells, glycolysis is inhibited under aerobic conditions. In the 1920s, German biochemist Otto Warburg discovered the famous "Warburg effect," which shows that even under sufficient oxygen, glycolysis is still active in malignant tumor cells, manifested by high glucose uptake and high levels of the metabolic product lactate. Therefore, the occurrence, growth, and transformation of cancer cells can utilize both mitochondrial oxidative phosphorylation and glycolysis to provide nutrients and energy. Tumor cells possess the ability to switch between these two energy metabolism pathways, a phenomenon known as metabolic variability, which leads to resistance to chemotherapy drugs.

[0004] Traditionally, nuclear DNA (nDNA) is considered the primary target of platinum-based antitumor drugs. Due to the existence of nDNA damage repair mechanisms within cells, damage to nDNA caused by platinum-based drugs is easily repaired, leading to drug resistance in tumor cells. Mitochondria contain circular mitochondrial DNA (mtDNA), which is the only genetic material in animal cells besides nDNA. Because mtDNA lacks histone protection and has relatively weak damage repair capabilities, it is more susceptible to damage and could become a potential target for platinum-based drugs. Therefore, targeting mtDNA with platinum-based complexes may affect the energy metabolism and survival ability of tumor cells, overcoming the drug resistance problem of traditional platinum-based antitumor drugs.

[0005] The role of tumor proteins and tumor suppressor proteins in promoting malignant transformation of mammalian cells by influencing properties such as proliferation signaling, cell cycle regulation, and adhesion alterations has been well-established. Chemicals, viruses, and radiation are also widely recognized as factors that typically lead to mutations in the genes encoding these oncogenic proteins, resulting in cancer. However, recent evidence suggests that two additional key factors influence proliferating cells during the transformation to malignant tumors: hypoxia and nutrient deprivation stress conditions (such as glucose deficiency). These additional triggers can initiate and promote the malignant transformation process when a low percentage of cells overcome and evade cellular senescence. It is increasingly evident that hypoxia leads to a progressive increase in mitochondrial ROS production (chronic ROS), which, over time, leads to cell stabilization through increased HIF-2alpha expression, enabling cells to survive under persistently elevated ROS levels. Evidence suggests that reactive oxygen species can mediate aberrant activity of human telomerase reverse transcriptase (hTERT) in cancer cells. Furthermore, hTERT can activate telomerase activity, maintain telomere length, prevent telomere degradation, and induce cell immortalization—all major hallmarks of cancer. In hypoxic or low-glucose cells, DNA mismatch repair is inhibited by HIF-2alpha, which continuously accumulates mitochondrial ROS-induced oxidative DNA damage and increases the number of mutations driving malignant transformation. Recent evidence also suggests that the resulting mutant oncogenes amplify this process through a combination of direct effects on mitochondrial function, which synergistically play a crucial role in a vicious cycle promoting malignant cell transformation. Therefore, malignant transformation of tumor cells is often the result of long-term damage to reactive oxygen species (ROS), toxic byproducts of oxidative phosphorylation, and mitochondria are a major source of ROS. Excessive ROS production promotes oncogene mutations. When mutant cells survive this oxidative stress, they may emerge as immortalized cells and initiate tumor progression.

[0006] Reactive oxygen species (ROS) are normal byproducts of cellular metabolism and are essential for many cellular biological functions. They also play a significant role in cancer development. Due to metabolic abnormalities, activation of oncogenic signals, and mitochondrial dysfunction, cancer cells produce excessive ROS, causing severe oxidative damage and ultimately leading to tumor cell death. Thioredoxin reductase (TrxR) is an important ROS scavenging enzyme overexpressed in various human tumors. It plays a crucial role in regulating intracellular redox homeostasis and protecting cancer cells from ROS-induced cell death. Therefore, mitochondrial TrxR has become a promising target for anticancer drug development.

[0007] Cisplatin and its analogues, such as carboplatin, oxaliplatin, nedaplatin, lobaplatin, and heptaplatin, have been approved for clinical use in various countries for the treatment of multi-faceted solid tumors, and approximately half of chemotherapy strategies involve platinum-based drugs. However, these drugs are structural homologs of cisplatin, and a drawback is that heritability, specifically DNA, is considered the ultimate target of cisplatin. The biggest problem with existing platinum-based anticancer drugs is drug resistance. This patent synthesizes a terpyridine platinum complex (TTP) that targets mitochondria. This compound, TTP, disrupts intracellular redox homeostasis by inhibiting thioredoxin reductase (TrxR) in mitochondria and the cytoplasm, damaging mitochondrial morphology and function. Simultaneously, it inhibits two energy metabolism pathways: mitochondrial oxidative phosphorylation and glycolysis, reducing the overall metabolic level of tumor cells, ultimately inhibiting tumor cell proliferation and overcoming cellular drug resistance. The mechanism of action of the compound TTP is as follows: Figure 1 As shown. Monofunctional PtII complexes are a potential novel metal drug that breaks the traditional structure-activity relationship of platinum-based drugs and exhibits high therapeutic efficacy. This application also conducted activity tests on the synthesized compound, which showed excellent performance and can serve as a very promising lead compound. Summary of the Invention

[0008] The purpose of this invention is to address the shortcomings of existing technologies by proposing a platinum complex metal antitumor drug TTP, its synthesis method, and its application. This metal antitumor drug breaks the traditional structure-activity relationship of platinum drugs and exhibits high therapeutic efficacy. The synthesized compound shows excellent activity in activity tests and can serve as a very promising lead compound.

[0009] To achieve the above objectives, the present invention employs the following technical solution: a platinum complex metal antitumor drug TTP, which is prepared by using 2-bromopyridine and 5-methyl-2-aldehydepyridine as raw materials, reacting n-butyllithium with bromopyridine for halolithium exchange, attacking pyridinecarboxaldehyde with an ion to obtain a hydroxydipyridine compound, further oxidizing it to obtain a carbodipyridine compound, undergoing NBS bromination, triphenylphosphine condensation, and potassium tetrachloroplatinate complexation, finally yielding the mitochondrial-targeting final product TTP; the general reaction formula is:

[0010] .

[0011] A method for synthesizing a platinum complex metal antitumor drug TTP, using 2-bromopyridine and 5-methyl-2-aldehydepyridine as starting materials, involves condensation, bromination, condensation, and complexation to finally obtain the mitochondrial-targeting final product TTP. The specific synthetic method includes the following steps:

[0012] Step 1: Hydroxydipyridine compound is synthesized from 2-bromopyridine and 5-methyl-2-aldehydepyridine. The reaction utilizes the reaction of n-butyllithium with bromopyridine to carry out halolithium exchange, and then the hydroxydipyridine compound is obtained by attacking pyridine formaldehyde with an ion. The compound is then further oxidized to obtain carbon-based dipyridine compound a.

[0013] Step 2: Substitute N-bromosuccinimide (NBS) with the methyl group on the pyridine ring to obtain the bromoproduct b;

[0014] Step 3: Add triphenylphosphine, and further substitution and elimination reactions yield triphenylphosphine compound d;

[0015] Step 4: Complex with potassium tetrachloroplatinate to obtain the final product, the e-metal antitumor drug TTP, which targets mitochondria.

[0016] The present invention also provides the application of TTP, a metal antitumor drug derived from a platinum complex obtained by the above-described synthesis method, in the preparation of antitumor active drugs.

[0017] Preferably, the tumor is non-small cell lung cancer A549.

[0018] By employing the above-mentioned technical solution, this invention disrupts intracellular redox homeostasis and damages mitochondrial morphology and function by inhibiting thioredoxin reductase (TrxR) in mitochondria and cytoplasm. Simultaneously, it inhibits two energy metabolism pathways—mitochondrial oxidative phosphorylation and glycolysis—reducing the overall metabolic level of tumor cells, ultimately achieving the goal of inhibiting tumor cell proliferation and overcoming cellular drug resistance. This monofunctional PtII complex is a potential novel metal drug, breaking the traditional structure-activity relationship of platinum drugs and exhibiting high therapeutic efficacy. Furthermore, this invention also conducted activity tests on the synthesized platinum complex TTP. This triple pyridine platinum complex (TTP) showed good inhibitory activity against non-small cell lung cancer (A549) in vitro, and exhibited high inhibitory activity against IC50 in A549 cells. 50 With a value of 5.17 µM, it exhibits excellent performance and can serve as a very promising lead compound.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] 1. This invention disrupts intracellular redox homeostasis and damages mitochondrial morphology and function by inhibiting thioredoxin reductase (TrxR) in mitochondria and cytoplasm. At the same time, it inhibits two energy metabolism pathways, mitochondrial oxidative phosphorylation and glycolysis, thereby reducing the overall metabolic level of tumor cells and ultimately achieving the goal of inhibiting tumor cell proliferation and overcoming cell drug resistance.

[0021] 2. The metal-based antitumor drug synthesized in this invention breaks the traditional structure-activity relationship of platinum-based drugs, exhibiting higher therapeutic efficacy. Activity testing of the synthesized platinum complex TTP showed that this triple pyridine platinum complex (TTP) exhibited good inhibitory activity against non-small cell lung cancer (A549) in vitro, and showed high efficacy against IC50 in A549 cells. 50 With a value of 5.17 µM, it exhibits excellent performance and can serve as a very promising lead compound.

[0022] 3. The synthesis method of the present invention uses inexpensive and readily available raw materials, has a novel structure, high yield, and simple separation and purification operations, making it suitable for mass production. The product exhibits good biological activity. Attached Figure Description

[0023] Figure 1 This is a schematic diagram illustrating the mechanism of action of the metal antitumor drug TTP of the present invention;

[0024] Figure 2 The proton NMR spectrum of compound a in this embodiment of the invention. Figure 1 H-NMR;

[0025] Figure 3 The above is the 1H NMR spectrum of triphenylphosphine dipyridine compound e(TTP) from an embodiment of the present invention. 1H-NMR;

[0026] Figure 4 The above is the 1H NMR spectrum of triphenylphosphine dipyridine compound e(TTP) from an embodiment of the present invention. 13 C-NMR;

[0027] Figure 5 This is a diagram illustrating the bioactivity of the triphenylphosphine dipyridine compound e (TTP) in non-small cell lung cancer A549 according to an embodiment of the present invention.

[0028] Figure 6 This is a diagram showing the mitochondrial membrane potential depolarization of the triphenylphosphine dipyridine compound e(TTP) in an embodiment of the present invention. Detailed Implementation

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, thereby making a clearer definition of the scope of protection of the present invention. The embodiments described in this invention are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0030] General methods

[0031] Compounds with delocalized positive charges and high lipophilicity can easily penetrate the inner mitochondrial membrane due to their high permeability. Triphenylphosphine (Ph3P) + Triphenylphosphine (TPP) is such a cation that can accumulate in mitochondria due to the negatively charged microenvironment within the mitochondrial matrix. Currently, TPP is widely used as a mitochondrial targeting agent, and TPP tethering agents have shown the potential to influence mitochondrial metabolism, thereby affecting the metabolism and apoptosis of cancer cells. For example, Ph3P... + Platinum-m, a cisplatin PtIV prodrug with axial ligand, is formulated into biocompatible polymer nanoparticles for mitochondrial targeted drug delivery, which can attack mtDNA and inhibit cisplatin-resistant cancers. This invention aims to synthesize the following product based on triphenylphosphine, with the target product shown in the following structural formula (compound c is an unstable intermediate):

[0032]

[0033] Unless otherwise specified, all reagents and solvents were commercially available and ready for use without further purification. Distilled water was used in all experiments. Subsequent responses were validated using thin-layer chromatography (TLC, UV 254nm) on silica gel plates (0.25mm, China Ltd.). Recordings were performed on a Bruker AV-500 MHz spectrometer. 1 H NMR (400 MHz) and 13 C10 NMR (100 MHz) spectra were recorded with solution and tetramethylsilane as internal standards. Chemical shifts are expressed in ppm, and coupling constants (J) are expressed in Hertz (Hz).

[0034] Example

[0035] Synthesis of carbon-based dipyridine a

[0036]

[0037] Add 35 ml of dried anhydrous tetrahydrofuran to a 50 ml three-necked flask, evacuate to an argon atmosphere to maintain a dry and oxygen-free reaction environment, and cool to -78°C using a dry ice-acetone bath. o At C, 316 mg of 2.0 mmol of 2-bromopyridine was added, followed by 2.4 mL of 1.0 mol / L n-butyllithium (n-BuLi). The reaction solution rapidly changed from colorless to red. Maintain at -78°C. o After reacting at C for 10 minutes, add 242 mg of 2.0 mmol of 5-methyl-2-aldehyde pyridine and maintain at -78°C. o The reaction mixture was reacted at C for 40 minutes, and then slowly heated to room temperature in a dry ice-acetone bath. The reaction was monitored by TLC. 40 mL of water was added, and excess tetrahydrofuran solution was removed by rotary evaporation. The reaction mixture was extracted with dichloromethane (DCM) (100 mL x 4), washed with saturated ammonium chloride (40 mL x 1) and brine (40 mL x 2), and dried over anhydrous sodium sulfate. The organic layer was concentrated by rotary evaporation under reduced pressure, followed by column chromatography to obtain the desired brown solid product.

[0038]

[0039]

[0040] A 50 mL three-necked flask was evacuated under argon protection. 35 mL of dried anhydrous tetrahydrofuran was added, followed by 198.2 mg (1.0 mmol) of brown compound a. Maintaining an anhydrous and oxygen-free reaction environment, 213.6 mg (1.2 mmol) of N-bromosuccinimide (NBS) was added. The reaction mixture was stirred at room temperature for 2 hours. TLC was used to monitor the reaction; once the starting material spot disappeared, 50 mL of water was added. The tetrahydrofuran was removed using a rotary evaporator. The reaction mixture was extracted with dichloromethane (DCM) (50 mL x 3), washed with saturated brine (brine) (50 mL x 2), and dried over anhydrous sodium sulfate for at least 1 hour. The organic layer was concentrated under reduced pressure to obtain the unpurified product b. Triphenylphosphine (341 mg, 1.2 mmol) and 30 mL of anhydrous 1,4-dioxane were added, and the reaction was carried out at room temperature for at least 2 hours. The reaction was monitored by TLC spotting. The TLC showed that the starting material disappeared and a highly polar spot appeared. 50 mL of water was added, and the solvent 1,4-dioxane was removed by rotary evaporation. The reaction mixture was extracted with dichloromethane (DCM) / isopropanol (iPrOH) (50 mL x 3) and dried with anhydrous sodium sulfate for at least 1 hour. The organic layer was concentrated under reduced pressure and purified by column chromatography to give product d (339.3 mg, yield 74%). Compound d (267.7 mg, 0.5 mmol) was dissolved in anhydrous and oxygen-free N,N-dimethylformamide (DMF, 15 mL). Under argon protection, potassium tetrachloroplatinate (415.1 mg, 1.0 mmol) was added, and the reaction was carried out in a dark chamber at 55 °C for 48 hours. The oily residue of the reaction solution was first washed with 50 mL of dichloromethane and extracted with hot methanol (200 mL). The extract was concentrated to 5 mL. After adding diethyl ether, a pale yellow precipitate was obtained. The ether was washed with dichloromethane and diethyl ether, and dried under vacuum to give the final product e (TTP, 329.6 mg, 4.55 mmol), with a yield of 91%.

[0041]

[0042] The specific experimental steps for the antitumor activity experiment of the antitumor compound TTP in this invention against human non-small cell lung cancer cells (A549) are as follows:

[0043] 1. Sample preparation: Weigh the sample accurately using the weight reduction method, dissolve it in DMSO, shake it to fully dissolve it, and prepare a 10 mmol / L stock solution. Aliquot and store the stock solution.

[0044] 2. Cell resuscitation and passage: Carefully remove the cryopreservation tube containing tumor cells from liquid nitrogen and thaw it rapidly in water at 37°C. Transfer the tumor cells to centrifuge tubes with culture medium (containing 95% basal medium, 5% newborn fetal bovine serum, and 1% penicillin-streptomycin antibiotics). After centrifugation, remove the supernatant and transfer the tumor cells to culture flasks containing culture medium. Incubate the flasks in an incubator (5% CO2, 37°C). Observe cell morphology and cell confluence under an inverted microscope. When the confluence reaches 70%~80%, remove the culture medium from the original culture flask for adherent cells (A549). Rinse the cells 1-2 times with PBS buffer. Remove the residual PBS with a pipette. Add 1 ml of trypsin containing EDTA for digestion (usually controlled at 1~2 min). Observe under the microscope. When the cell edges shrink and some adherent cells begin to move (it is not recommended to digest until the cells float), remove the trypsin. Gently tap the culture flask. When a large number of cells slip off, add culture medium to stop digestion. Gently blow the cell layer to disperse the cell layer as much as possible. Transfer to a centrifuge tube and centrifuge. After centrifugation, remove the supernatant. Use culture medium to disperse the cells into single cells as much as possible and transfer to a new culture flask. Place in an incubator for further culture. If it is suspension cells, skip the digestion process and centrifuge directly. The operation procedure is the same as above.

[0045] 3. Cell Plating: For cells in the logarithmic growth phase, perform the same steps as in (1). Mix 20 µL of cell suspension with 80 µL of PBS. Mix 90 µL of cell suspension with 10 µL of trypan blue staining agent. Transfer 10 µL of the diluted cell suspension onto a hemocytometer. Count the number of cells in four squares as n. The cell count / mL = n / 4 dilution factor. The plating concentration is 1×10⁻⁶. 5 If y cells are needed for plating, the required cell suspension volume v = y / number of cells per milliliter. Take a certain volume of cell suspension and add complete culture medium to the required volume, then thoroughly pipette the cells evenly. Leave the outer edge wells of the 96 plate empty initially, and add 100 µL of diluted cell suspension to each of the remaining wells. Incubate in an incubator for 24 h.

[0046] 4. Sample addition: Dilute the stock solution with basal culture medium in a gradient manner to prepare sample solutions of 50, 25, 12.5, 6.25, 3.12, 1.55, and 0.78 µmol / L. Perform parallel experiments with three replicates for each sample and concentration, and include a control group (no drug added) and a positive drug group. Add 10 µL of sample solution to the wells containing cells, and add 100 µL of PBS to each of the peripheral wells. Incubate for 72 hours.

[0047] 5. Measure OD value: Add 10µL of MTS solution to each well and continue culturing. Stop incubation when the OD value of the untreated group differs from that of the treated group by more than 3 times, and measure the OD value of each well at a wavelength of 490 nm using a microplate reader.

[0048] ;

[0049] Based on the inhibition rate results, data processing was performed using GraphPad Prism 5.0 software to calculate the half-maximal inhibitory concentration (IC50) for the proliferation of tumor cells and normal cell lines. 50 This triple pyridine platinum complex (TTP) exhibited good inhibitory activity against non-small cell lung cancer (A549) in vitro, and showed good inhibitory activity against the IC50 of A549 cells. 50 The value is 5.17 µM ( Figure 5 ), among which, such as Figure 5 As shown, the cell growth status is different at different concentrations: when the non-small cell lung cancer (A549) cells are at the leftmost (Contr) level, the cells are still growing normally; at the middle levels (0.78, 1.55, and 3.12 uM / L), the cells show severe apoptosis; and at the rightmost level... Figure 6 The 0.25uM / L level has almost completely resulted in apoptosis.

[0050] In the early stages of apoptosis, mitochondrial membrane permeability increases, and the mitochondrial membrane potential decreases or even disappears. Therefore, changes in mitochondrial membrane potential can reflect the level of apoptosis. JC-1 is a commonly used fluorescent probe for detecting changes in mitochondrial membrane potential. When the mitochondrial membrane potential is high, JC-1 accumulates in the mitochondrial matrix, forming red fluorescence; when the mitochondrial membrane potential is low, JC-1 cannot accumulate in the mitochondrial matrix and exhibits green fluorescence. Figure 6 ).

[0051] The experimental steps are as follows:

[0052] 1. Cell preparation (adherent cells): Transfer the cells to a confocal dish and gently shake until homogeneous, then culture until 80-90% homogeneous.

[0053] 2. Preparation of JC-1 staining working solution: Take 50 μL of JC-1 (200×) and add 8 mL of PBS to dilute JC-1. Then add 2 mL of JC-1 staining buffer (5×) and mix well to obtain the JC-1 staining working solution.

[0054] 3. Washing: Discard the culture medium and wash 2-3 times with PBS.

[0055] 4. Staining: Add 1 mL of JC-1 staining working solution and mix thoroughly.

[0056] 5. Incubation: Incubate at 37°C for 20 minutes in a cell culture incubator.

[0057] 7. Washing: After incubation, discard the supernatant, wash 2-3 times with JC-1 staining buffer (1×), and then add 2mL of PBS or culture medium (which may contain serum).

[0058] 8. Observation and Photography: Observe under a fluorescence microscope or laser confocal microscope. The excitation wavelength is usually 488 nm, and the emission wavelengths are 590 nm for red fluorescence and 530 nm for green fluorescence. When the level of apoptosis decreases, the mitochondrial membrane potential increases, i.e., the red fluorescence is stronger; when the level of apoptosis increases, the mitochondrial membrane potential decreases, i.e., the red fluorescence weakens, or green fluorescence appears.

[0059] JC-1 staining, a method based on membrane permeability, is widely used in apoptosis studies to monitor mitochondria. JC-1 dye can be used as an indicator of mitochondrial membrane potential in various cell types (including muscle cells and neurons), as well as in intact tissues and isolated mitochondria. Figure 6 As shown, the left image is the red group without TTP, the right image is the group with TTP, and the green image is the group with TTP, indicating the collapse of mitochondrial membrane potential.

[0060] In summary, the synthesis method of this invention uses inexpensive and readily available raw materials, has a novel structure, high yield, and simple separation and purification operations, making it suitable for large-scale production. The product exhibits good biological activity, and detection shows that the platinum complex metal antitumor drug TTP has an IC50 of [missing information - likely related to a specific target or effect]. 50 It is 5.17uM.

[0061] The descriptions and practices disclosed in this invention are readily apparent and understandable to those skilled in the art, and various modifications and refinements can be made without departing from the principles of this invention. Therefore, any modifications or improvements made without departing from the spirit of this invention should also be considered within the scope of protection of this invention.

Claims

1. A platinum complex metal antitumor drug TTP, characterized in that, Using 2-bromopyridine and 5-methyl-2-aldehydepyridine as starting materials, a halide lithium exchange reaction is carried out via n-butyllithium and bromopyridine. The pyridine carboxaldehyde is attacked by anion to obtain a hydroxydipyridine compound, which is further oxidized to obtain a carbodipyridine compound. This compound is then subjected to NBS bromination, triphenylphosphine condensation, and potassium tetrachloroplatinate complexation to finally yield the mitochondrial-targeted final product, TTP. The general reaction formula is: 。 2. The method for synthesizing the platinum complex metal antitumor drug TTP according to claim 1, characterized in that, Using 2-bromopyridine and 5-methyl-2-aldehydepyridine as starting materials, the synthesis proceeds through condensation, bromination, condensation, and complexation to finally obtain the mitochondrial-targeted final product TTP. The specific synthetic method includes the following steps: Step 1: Hydroxydipyridine compound is synthesized from 2-bromopyridine and 5-methyl-2-aldehydepyridine. The reaction utilizes the reaction of n-butyllithium with bromopyridine to carry out halolithium exchange, and then the hydroxydipyridine compound is obtained by attacking pyridine formaldehyde with an ion. The compound is then further oxidized to obtain carbon-based dipyridine compound a. Step 2: Substitute N-bromosuccinimide (NBS) with the methyl group on the pyridine ring to obtain the bromoproduct b; Step 3: Add triphenylphosphine, and further substitution and elimination reactions yield triphenylphosphine compound d; Step 4: Complex with potassium tetrachloroplatinate to obtain the final product, the e-metal antitumor drug TTP, which targets mitochondria.

3. The application of TTP, a metal antitumor drug derived from a platinum complex obtained by the synthesis method described in claim 2, in the preparation of antitumor active drugs.

4. The application according to claim 3, characterized in that, The tumor is non-small cell lung cancer A549.

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