Novel targeting compound for treating various cancer types as well as preparation method and application of novel targeting compound

Abstract

The invention belongs to the technical field of organic arsine targeting compounds, and particularly relates to a novel targeting compound for treating various cancers as well as a preparation method and application of the novel targeting compound. Core innovations include precise synthesis of an arsine precursor and a process for designing a targeting structure for multiple cancer species. A series of high-purity organic arsine compounds are developed through an advanced organic synthesis technology, and screening optimization is carried out in combination with means such as computational simulation, biomacromolecule interaction analysis, cell experiments and animal models. Results show that the compound has the characteristics of efficient tumor inhibition and low toxicity, especially shows strong inhibition on thioredoxin reductase, prolongs the in-vivo circulation time and reduces the administration frequency. The effect in treatment of the triple-negative breast cancer is remarkable, and the good clinical transformation potential is achieved. The invention aims to protect the chemical structure and the preparation method of the compound and the application of the compound in cancer treatment.

Description

Technical Field

[0001] The present invention relates to the field of organic arsenic targeting compounds, and specifically discloses a novel targeting compound for treating various cancer types, a preparation method thereof, and an application thereof. Background Art

[0002] In the field of cancer therapy, arsenic agents, such as arsenic trioxide (ATO), GSAO, and ZIO101, have achieved breakthroughs in hematologic malignancies (such as acute promyelocytic leukemia). However, their application in solid tumors still faces common challenges: low activity leading to dose-dependent toxicities (such as liver and kidney damage and cardiotoxicity), rapid metabolism leading to insufficient exposure, and the nonspecific distribution defects of traditional chemotherapeutic agents. To address these challenges, novel organoarsine compounds have been innovatively designed to overcome tumor type limitations. Their core mechanistic target, thioredoxin reductase (TrxR), is highly expressed in various solid tumors, including liver cancer, non-small cell lung cancer, and colorectal cancer, and is closely associated with tumor proliferation, metastasis, and drug resistance. By constructing a "TrxR-oxidative stress" dual-axis regulatory system, organoarsines covalently bind to the selenocysteine residue in the active center of TrxR, inhibiting the antioxidant system while inducing a burst of reactive oxygen species (ROS). This mechanism has demonstrated cross-tumor killing effects in preclinical models of breast cancer, pancreatic cancer, and glioma.

[0003] To realize the broad-spectrum anti-cancer potential, new organoarsine compounds break through the limitations of traditional drugs through multi-dimensional structural optimization strategies:

[0004] 1. High efficiency: single administration achieves deep treatment

[0005] By inhibiting key enzymes in drug metabolism, the drug's circulation time in the body is significantly prolonged, increasing sustained exposure within tumor tissue. Combined with liposome drug delivery technology to enhance membrane permeability and tumor-targeted enrichment, a single dose can achieve complete elimination of tumor cells, overcoming the drawbacks of traditional chemotherapy, which requires frequent dosing.

[0006] 2. Low toxicity: Precise targeting reduces systemic damage

[0007] Based on a tumor microenvironment-specific activation mechanism, the drug selectively releases its active ingredients within cancer cells, significantly reducing nonspecific damage to normal tissues. By targeting and regulating the tumor antioxidant system and oxidative stress pathways, it can effectively kill tumor cells at extremely low concentrations, significantly reducing the risk of toxicity to key organs such as the liver, kidneys, and heart.

[0008] 3. Low cost: scalable industrial production path

[0009] Using modular molecular design and a streamlined synthesis process, functional modifications are achieved with high yields, avoiding complex purification steps. The readily available and cost-effective raw materials, combined with the potential for large-scale production, provide the drug with universal economic advantages comparable to traditional chemotherapy, laying the foundation for widespread clinical application.

[0010] By targeting the specific target TrxR protein structure, the same drug can achieve specific activation in different cancer types, such as breast cancer, leukemia, and cervical cancer. This "one drug, multiple functions" strategy broadens the coverage of treatment scenarios while maintaining low toxicity. Summary of the Invention

[0011] In response to the shortcomings of the prior art, the present invention aims to provide a novel targeted compound for treating multiple types of cancer, a preparation method thereof, and an application thereof. The present invention discloses a novel targeted compound for treating multiple types of cancer, and a preparation method thereof. Through innovative organic synthesis, a variety of organic arsenic compounds are prepared. Through computational simulation, screening and exploration at the levels of macromolecules, cells, and small animals, a series of highly effective and low-toxic targeted anti-tumor organic arsenic compounds are obtained. The single-component organic arsenic in this series has higher thioredoxin reductase inhibition ability and longer in vivo circulation half-life. Under the action of lower dosage and lower dosage frequency, targeted inhibition of tumors is achieved, and the inhibitory effect on multiple types of cancer is very significant, which has practical significance.

[0012] To achieve the above object, the present invention provides the following technical solutions:

[0013] A novel targeted compound for treating various cancer types, wherein the molecular structure of the organic arsenic targeted compound is:

[0014]

[0015] Wherein, the R1 is one of nitro and n-butoxy;

[0016] R2 is one or more methoxy groups.

[0017] Preferably, the alkyl group includes methyl (—CH3), ethyl (—C2H5), n-propyl (—C3H7), isopropyl (—C3H7), n-butyl (—C4H9), isobutyl (—C4H9), tert-butyl (—C4H9), pentyl (—C5H 11 ) etc. until dodecyl (—C 12 H 25); Olefins: vinyl (—CH=CH2), propenyl (—CH2CH=CH2), butenyl (—C4H7), etc., including cis and trans isomers; Alkoxy: methoxy (—OCH3), ethoxy (—OC3H5), n-propoxy (—OC3H7), isopropoxy (—OC3H7), n-butoxy (—OC4H9), etc. up to dodecyloxy (—OC 12 H 25 ); Alkylamino: methylamino (—NHCH3), ethylamino (—NHC2H5), dimethylamino (—N(CH3)2), diethylamino (—N(C2H5)2), etc.; Aminoalkyl: one or more of aminomethyl (—CH2NH2), 2-aminoethyl (—CH2CH2NH2), and 3-aminopropyl (—CH2CH2CH2NH2).

[0018] Preferably, the arylalkylphenyl group includes tolyl, ethylphenyl, etc.; nitrophenyl group includes p-nitrophenyl, o-nitrophenyl, m-nitrophenyl; hydroxyphenyl group includes p-hydroxyphenyl, o-hydroxyphenyl, m-hydroxyphenyl; halogenated phenyl group includes fluorophenyl, chlorophenyl, bromophenyl, iodophenyl (each halogen can be located at different positions of the benzene ring to form different isomers); one or more of naphthyl, anthracenyl, and biphenyl.

[0019] The heterocyclic substituents are oxirane, oxirane, oxirane, acridinyl, pyrrolidinyl, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, furanyl, thiophene, piperidinyl, pyridyl, morpholinyl, piperazinyl, aza Oxy or thia One or more of;

[0020] The halogen atom is one or more of fluorine, chlorine, bromine and iodine.

[0021] A method for preparing a novel targeted compound for treating multiple cancer types, used to prepare the triple-negative breast cancer targeted compound, the preparation method is as follows:

[0022] Preparation of S1 Arsine Precursor:

[0023] 1. Preparation and heating of reaction medium: First, accurately measure 8-12 ml of 70% ammonium thioglycolate solution (13 g, 120 mM) and place it in an appropriate reaction vessel. Use a heating device to control the solution temperature at 50°C to ensure the smooth progress of the subsequent reaction.

[0024] 2. Gradually add the reactants: While stirring continuously and maintaining the temperature, slowly add 3.5-4.5 g (23 mM) of phenylarsonic acid 1a and 1b to the hot solution in batches (about 4-5 times). This step takes about 1 hour to ensure a smooth reaction and avoid violent exotherm or local overconcentration.

[0025] 3. Add the second reactant: After about 1 hour, use a constant pressure dropping funnel to control the rate and slowly add 5 ml (29 mM) of 1,3-propanedithiol to the reaction mixture. This operation is carried out under mild conditions to ensure the gradual progress of the reaction and efficient coupling.

[0026] 4. Reaction and Post-Processing: The entire reaction system was placed in an oil bath and heated for 4 hours to allow the reactants to fully react. After the reaction, the resulting suspension was extracted with dichloromethane in multiple small amounts to effectively extract the target product into the organic phase. After extraction, all organic layers were combined and concentrated using a rotary evaporator to remove the solvent.

[0027] 5. Purification and Product Collection: Finally, the concentrated crude product is separated and purified by silica gel column chromatography, using a mixed solvent of petroleum ether (PE) and dichloromethane (DCM) in a 2:1 volume ratio as the eluent to achieve optimal separation. By monitoring the physical and chemical properties of the effluent during column chromatography, pure fractions of the target product are collected, completing the synthesis and purification of the organoarsine precursor.

[0028] This synthetic strategy ensures efficient synthesis and high-purity recovery of the target product through precise operational control and optimized post-processing steps, providing high-quality organoarsine compound precursor samples for subsequent chemical characterization and biological activity testing.

[0029] Synthesis of organic arsenic compounds: The organic arsenic targeting compound is prepared by mixing the arsenic precursors 2a and 2b prepared in S1 with raw material compounds for reaction.

[0030] Preferably, the synthesis steps of the organoarsine targeting compound are as follows:

[0031] S101. A series of drugs containing 1 mM P450 inhibitory structures were dissolved in 10 ml of dichloromethane (DCM) solvent to ensure uniform dispersion of the drug molecules. Subsequently, 3-4 drops of N,N-dimethylformamide (DMF) were added as a catalyst to promote the smooth progress of subsequent reactions. 300 μl of oxalyl chloride was pre-dissolved in 15 ml of dichloromethane to improve the solubility of the reaction raw materials and the uniformity of the reaction. Using a constant pressure dropping funnel technique, this oxalyl chloride solution was slowly and evenly added dropwise to the reaction mixture in a single-necked flask. This reaction is expected to produce chlorine gas. To ensure laboratory safety and prevent environmental contamination, a gas recovery system was specially installed above the constant pressure funnel to effectively capture and treat the generated harmful gases. Under strictly controlled conditions, the reaction was continued for 1 hour, during which the reaction progress was closely monitored. After the reaction was completed, the solvent was removed by rotary evaporation to obtain the expected oily product.

[0032] S102. Under room temperature, the experimental procedure was carried out as follows to synthesize the target compound: First, 1 mM of the drug acyl chloride structure that inhibits the P450 enzyme structure was dissolved in 10 ml of dichloromethane (DCM), ensuring complete dissolution to form a uniform reaction substrate solution. Next, 273 mg (1 mM) of compound Z2 was dissolved in 15 ml of DCM and slowly added to the solution containing the monosubstituted phenyl acyl chloride at a constant speed and steady rate using a constant pressure dropping funnel. This step ensured the gradual progress of the reaction and efficient coupling.

[0033] 300 μl of triethylamine was added to the system as an effective acid-binding agent to neutralize any acidic byproducts that might be produced during the reaction, thereby promoting the formation of the desired product. To monitor the reaction progress in real time, thin-layer chromatography (TLC) was used periodically until the TLC results showed the near-complete disappearance of the monosubstituted phenylacyl chloride spot, indicating near-completion of the reaction. This process typically lasted 2-3 hours.

[0034] After the reaction is complete, the product is washed once with 10% hydrochloric acid to remove any alkaline impurities, then washed once with saturated sodium bicarbonate (NaHCO3) solution to neutralize any residual acidic substances, and finally washed once with pure water to remove inorganic salts and other water-soluble impurities. After washing, the organic phase is dried over anhydrous sodium sulfate (Na2SO4) to remove residual water and ensure product purity.

[0035] Finally, the pretreated organic phase was purified by silica gel column chromatography using a 2:1 volume ratio of dichloromethane (DCM) to petroleum ether (PE) as the eluent. Optimized separation conditions allowed for high-resolution separation and collection of the target product, resulting in high-purity, desired compounds. This method integrates meticulous operational control with an efficient purification strategy to ensure high product quality and yield.

[0036] The preparation routes and structures of organoarsine targeting compounds 3c and 3d are as follows:

[0037]

[0038] Among them, taking the organic arsenic targeted drug 3c-3d as an example, in the organic arsenic targeted compound 3c, the position of 2-methyl-1,3,2-dithiarsine is 4; in the organic arsenic targeted compound 3d, the position of 2-methyl-1,3,2-dithiarsine is 2.

[0039] Application of an organoarsine targeting compound prepared according to the organoarsine targeting compound or the preparation method of the organoarsine targeting compound in treating various cancer types.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] 1. Integrated design of metabolic regulation and long-term treatment

[0042] By precisely introducing a P450 enzyme metabolism inhibitory group into an organic arsenic precursor molecule, the short-lived efficacy of traditional arsenic agents due to rapid metabolism is overcome, significantly extending the drug's circulation time in the body. The resulting single-component compound maintains high anti-tumor activity while effectively avoiding the hepatotoxicity, renal toxicity, and adverse cardiac reactions associated with traditional arsenic agents (such as arsenic trioxide) by reducing dosing frequency and single dose, achieving dual optimization of efficacy and safety.

[0043] 2. Pioneering breakthroughs in the TrxR target across multiple cancer types

[0044] This is the first organic arsenic compound to precisely target thioredoxin reductase (TrxR). Compared to the similarly targeted drug auranofin (which requires high doses and is not indicated for cancer) and the broad-spectrum arsenic agent arsenic trioxide, this compound, through the synergistic effects of targeted activation and reversal of metabolic resistance, can specifically inhibit the proliferation and metastasis of various cancer cells at extremely low doses, providing the technical foundation for the first targeted therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 1. Multiple process flow charts for the preparation of the organoarsine targeting compound of the present invention;

[0046] Figure 2 The synthesis process of the precursor of the organic arsenic target compound of the present invention;

[0047] Figure 3 This is the synthesis process of the acyl chloride in the organic arsenic target compound of the present invention;

[0048] Figure 4This is the synthesis process of the target compound in the organoarsine target compound of the present invention;

[0049] Figure 5 This is a diagram for detecting the TrxR activity of the organoarsine targeting compound of the present invention:

[0050] Figure 6 This is a fluorescence signal detection diagram of the organoarsine targeting compound of the present invention;

[0051] Figure 7 This is a detection diagram of the main active oxygen species of the organoarsenic targeting compound of the present invention;

[0052] Figure 8 This is a confocal detection image of the organoarsine targeting compound of the present invention;

[0053] Figure 9 This is a dual-channel fluorescence signal detection diagram of the organoarsine targeting compound of the present invention;

[0054] Figure 10 Pharmacokinetic experimental detection diagrams of the organoarsenic targeting compound of the present invention: Figure A is a pharmacokinetic experimental operation flow chart; Figure B is a curve chart showing the change of arsenic concentration in blood over time. DETAILED DESCRIPTION

[0055] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0056] See also Figure 1-10 , the present invention provides a technical solution:

[0057] An organic arsenic targeting compound, the molecular structure of the organic arsenic targeting compound is:

[0058]

[0059] Wherein, the R1 is one of nitro and n-butoxy;

[0060] R2 is one or more methoxy groups.

[0061] The alkyl groups: methyl (—CH3), ethyl (—C2H5), n-propyl (—C3H7), isopropyl (—C3H7), n-butyl (—C4H9), isobutyl (—C4H9), tert-butyl (—C4H9), pentyl (—C5H 11 ) etc. until dodecyl (—C 12 H 25); olefins: vinyl (—CH=CH2), propenyl (—CH2CH=CH2), butenyl (—C4H7), etc., including cis and trans isomers; alkoxy: methoxy (—OCH3), ethoxy (—OC2H5), n-propoxy (—OC3H7), isopropoxy (—OC3H7), n-butoxy (—OC4H9), etc. up to dodecyloxy (—OC 12 H 25 ); Alkylamino: methylamino (—NHCH3), ethylamino (—NHC2H5), dimethylamino (—N(CH3)2), diethylamino (—N(C2H5)2), etc.; Aminoalkyl: one or more of aminomethyl (—CH2NH2), 2-aminoethyl (—CH2CH2NH2), and 3-aminopropyl (—CH2CH2CH2NH2).

[0062] The arylalkylphenyl group includes tolyl, ethylphenyl, etc.; nitrophenyl group includes p-nitrophenyl, o-nitrophenyl, m-nitrophenyl; hydroxyphenyl group includes p-hydroxyphenyl, o-hydroxyphenyl, m-hydroxyphenyl; halogenated phenyl group includes fluorophenyl, chlorophenyl, bromophenyl, iodophenyl (each halogen can be located at different positions of the benzene ring to form different isomers); one or more of naphthyl, anthracenyl, and biphenyl.

[0063] The heterocyclic substituents are oxirane, oxirane, oxirane, acridinyl, pyrrolidinyl, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, furanyl, thiophene, piperidinyl, pyridyl, morpholinyl, piperazinyl, aza Oxy or thia One or more of;

[0064] The halogen atom is one or more of fluorine, chlorine, bromine and iodine.

[0065] In the present invention, the molecular formula of the organic arsenic targeting compound is disclosed in accordance with the content of the following specific embodiments, and specifically discloses but is not limited to the molecular formula of the following compounds:

[0066]

[0067] Regarding the preparation method of the above-mentioned organoarsine targeting compound, the present invention further discloses the following specific examples.

[0068] A method for preparing a synthetic organic arsenic targeting compound, taking compound 3c as an example, comprises the following steps:

[0069] S101 takes the following steps in the synthesis process of preparing an organic arsenic compound precursor:

[0070] 1. Preparation and heating of reaction medium: First, accurately measure 10 ml of 70% ammonium thioglycolate solution (13 g, 120 mM) and place it in an appropriate reaction vessel. Use a heating device to control the solution temperature at 50°C to ensure the smooth progress of the subsequent reaction.

[0071] 2. Gradually add the reactants: While stirring continuously and maintaining the temperature, slowly add 5 g (23 mM) of phenylarsonic acid 1a and 1b to the hot solution in batches (about 4-5 times). This step takes about 1 hour to ensure a smooth reaction and avoid violent exotherm or local overconcentration.

[0072] 3. Add the second reactant: After about 1 hour, use a constant pressure dropping funnel to control the rate and slowly add 5 ml (29 mM) of 1,3-propanedithiol to the reaction mixture. This operation is carried out under mild conditions to ensure the gradual progress of the reaction and efficient coupling.

[0073] 4. Reaction and Post-Processing: The entire reaction system was placed in an oil bath and heated for 4 hours to allow the reactants to fully react. After the reaction, the resulting suspension was extracted with dichloromethane in multiple small amounts to effectively extract the target product into the organic phase. After extraction, all organic layers were combined and concentrated using a rotary evaporator to remove the solvent.

[0074] 5. Purification and product collection: Finally, the concentrated crude product was separated and purified by silica gel column chromatography, using a mixed solvent of petroleum ether (PE) and dichloromethane (DCM) with a volume ratio of 2:1 as the eluent to achieve a good separation effect. By monitoring the physical and chemical properties of the effluent during column chromatography, the pure fractions of the target product were collected to complete the synthesis and purification process of the organic arsenic precursor 2a. The experimental process is as follows: Figure 2 shown.

[0075] This synthetic strategy ensures efficient synthesis and high-purity recovery of the target product through precise operational control and optimized post-processing steps, providing high-quality organic arsenic precursor 2a samples for subsequent chemical characterization and biological activity testing.

[0076] Synthesis of S102 p-Nitrophenylethyl Chloride

[0077] A suitable single-necked flask was selected as the reaction vessel. First, a series of drugs containing 1 mM p-nitrophenylethyl groups were dissolved in 10 ml of dichloromethane (DCM) to ensure uniform dispersion of the drug molecules. Subsequently, 3-4 drops of N,N-dimethylformamide (DMF) were added as a catalyst to facilitate the subsequent reactions.

[0078] Predissolve 300 μl of oxalyl chloride in 15 ml of dichloromethane to improve the solubility of the starting materials and ensure uniform reaction. Using a constant-pressure dropping funnel, slowly and evenly add this oxalyl chloride solution to the reaction mixture in a single-necked flask. This reaction is expected to produce chlorine gas. To ensure laboratory safety and prevent environmental contamination, a gas recovery system was installed above the constant-pressure funnel to effectively capture and treat the generated hazardous gas.

[0079] Under strictly controlled conditions, the reaction was continued for 1 hour, during which the reaction progress was closely monitored. After the reaction was completed, the solvent was removed by rotary evaporation to obtain the expected oily product. Figure 3 shown.

[0080] Synthesis of S103 target product

[0081] The target compound was synthesized under room temperature using the following procedures: First, 1 mM p-nitrophenylethyl chloride was dissolved in 10 ml of dichloromethane (DCM), ensuring complete dissolution to form a uniform reaction substrate solution. Next, 273 mg (1 mM) of an organoarsine precursor compound was dissolved in 15 ml of DCM. This organoarsine precursor solution was slowly added to the p-nitrophenylethyl chloride solution at a constant pressure dropwise rate using a constant pressure dropping funnel. This step ensured a gradual reaction and efficient coupling.

[0082] 300 μl of triethylamine was added to the system as an effective acid-binding agent to neutralize any acidic byproducts that may be produced during the reaction, thereby promoting the formation of the desired product. To monitor the reaction progress in real time, thin-layer chromatography (TLC) was used periodically until the TLC results showed the near-complete disappearance of the p-nitrophenylethyl chloride spot, indicating near-completion of the reaction. This process typically lasted 2-3 hours.

[0083] After the reaction is complete, the product is washed once with 10% hydrochloric acid to remove any alkaline impurities, then washed once with saturated sodium bicarbonate (NaHCO3) solution to neutralize any residual acidic substances, and finally washed once with pure water to remove inorganic salts and other water-soluble impurities. After washing, the organic phase is dried over anhydrous sodium sulfate (Na2SO4) to remove residual water and ensure product purity.

[0084] Finally, the pretreated organic phase was purified by silica gel column chromatography, using a mixed solvent of dichloromethane (DCM) and petroleum ether (PE) with a volume ratio of 2:1 as the eluent. The separation conditions were optimized, and the target product was separated and collected at high resolution, thereby obtaining the desired compound with high purity. This method integrates precise operational control with an efficient purification strategy to ensure the quality and yield of the synthesized product. The experimental process is as follows Figure 4 shown.

[0085] 1. MTT assay to detect the anti-proliferative ability of organic arsenic

[0086] Experimental plan: Cells (molm-13, NB4, 4T1, MCF-7, Hela, COS-7) were seeded into 96-well plates and cultured overnight to restore growth. Different concentrations of organic arsenic were added and cultured for 24 or 48 hours. After incubation with MTT, the cells were lysed and the absorbance at 490 nm was measured. The IC50 value was calculated. The results are shown in Table 1:

[0087] Table 1. IC values of organoarsenic compounds against cancer cell lines 50 value

[0088]

[0089] Data Analysis:

[0090] Among all organoarsines, organoarsine 5c has the lowest half-inhibitory concentration (CI) against triple-negative breast cancer 4T1 cells, the highest CI against normal COS-7 cells, and the highest IC50 ratio for normal cells to cancer cells (last column in the table). This indicates that organoarsine 5c has high anti-tumor activity, low ability to interfere with normal cell proliferation, and high selectivity for cancer cells versus normal cells. Because 4T1 cells lack clinically available targeted drug therapies, this cancer cell type was selected for subsequent biological studies.

[0091] 2. Targeted inhibition ability of organoarsenic on target TrxR

[0092] Experimental plan: Optimize the structure of organic arsenide using Gaussian; perform molecular docking of organic arsenide and TrxR using ledock and calculate the binding energy. Δ G.

[0093] 4T1 cells were plated in 10 cm culture dishes and cultured overnight. 0.9 μM organic arsenic was added and incubated for 24 hours. The cells were then harvested, homogenized, and lysed. Insoluble matter was removed by centrifugation (8000 g, 4°C, 10 min). TrxR activity in the supernatant was then assayed using a kit. The results are shown in Table 2.

[0094] Table 2. Interactions between organic arsenides and TrxR

[0095]

[0096] Data Analysis:

[0097] Molecular docking experiments revealed that the series of organoarsine compounds designed in this invention bind significantly better to thioredoxin reductase (TrxR) than their arsenic precursors 2a and 2b. Of particular note, organoarsine 5c exhibits the largest absolute value of the Gibbs free energy change (ΔG) upon binding to TrxR, indicating that it possesses the strongest affinity for TrxR among the series.

[0098] Further in vitro holoenzyme activity assays revealed that organoarsine 5c exhibited the lowest half-inhibitory concentration (IC50) for TrxR, indicating its most potent inhibitory potency against TrxR. Furthermore, under the premise of controlling the concentration of all organoarsines, the lowest TrxR activity was observed in the organoarsine 5c-treated group. This finding reveals that organoarsine 5c not only exhibits excellent TrxR inhibitory ability at the cellular level, but also confirms its great potential as a highly effective TrxR inhibitor.

[0099] 3. Oxidative stress and mitochondrial damage induce cell apoptosis

[0100] Experimental plan: 4T1 cells were seeded into 10 cm dishes. After overnight culture, organic arsenic was added and incubated for 3, 6, 9, 12, and 24 hours. DTNB was added. TrxR catalyzed the reduction of DTNB by NADPH to generate TNB and NADP+. TNB has a characteristic absorption peak at 412 nm. The fluorescence signal was detected by a microplate reader. The detection results are as follows: Figure 5 shown.

[0101] 4T1 cells were seeded into 6-well plates and cultured overnight. Arsenic was added and incubated for 3, 6, 12, and 24 hours. Reactive oxygen species (ROS) detection reagent DCFH-DA was added and incubated for half an hour. The cells were collected and the fluorescence signal was detected by flow cytometry. The detection results were as follows: Figure 6 shown.

[0102] 4T1 cells were seeded into 10 cm dishes and cultured overnight. Arsenic was added and incubated for 3, 6, 9, 12, and 24 hours. Divalent iron ions were added according to the kit to detect the changes in H2O2 levels. The test results were as follows: Figure 7 shown.

[0103] 4T1 cells were seeded into 1 cm confocal microplates and cultured overnight. Arsine was added and incubated for 3, 6, 9, 12, and 24 h. JC-1 dye was added according to the kit for confocal microscopy. The results were as follows: Figure 8 shown.

[0104] 4T1 cells were seeded into 6-well plates, cultured overnight, and then incubated with organic arsenic 5c for 24 hours. Apoptosis detection reagent was added and incubated. Cells were collected and dual-channel fluorescence signals were detected by flow cytometry. The detection results were as follows: Figure 9 shown.

[0105] Data Analysis:

[0106] like Figure 5 As shown, the results of microplate reader analysis showed that organic arsenic 5c could inhibit the TrxR level of 4T1 cells, resulting in a decrease in enzyme activity.

[0107] like Figure 6 As shown, flow cytometric analysis results showed that organoarsine 5c could induce elevated levels of reactive oxygen species in 4T1 cells.

[0108] like Figure 7 The results showed that organoarsine 5c induced an increase in the level of reactive oxygen species in cells. The main reactive oxygen species was H2O2, indicating that organoarsine 5c inhibited the activity of TrxR, induced oxidative stress and endoplasmic reticulum stress in cells, and further induced cell apoptosis.

[0109] like Figure 8 As shown, the confocal microscopy results after cell fixation showed that after incubation with organic arsenic 5c, the mitochondrial structure of the cells was destroyed, such as the reduction of content and destruction of the inner membrane, which led to changes in the potential of the mitochondrial membrane.

[0110] like Figure 9 As shown, organoarsine 5c induces oxidative stress in cells, induces endoplasmic reticulum stress, disrupts the structure and function of organelles such as mitochondria, and further induces cell apoptosis. Compared with the control group (C), cells incubated with organoarsine 5c at a concentration of 2×IC50 only had 23.5% of normal cells, while the rest underwent apoptosis.

[0111] 4.5c inhibits the activity of microsomal enzymes, prolongs the half-life of circulation in the body, and enhances anti-tumor activity

[0112] Adult healthy male SD rats were selected and injected with 1.5 mg / kg of organic arsenic via the tail vein. Blood samples were collected from the rats at the predetermined time points of 0, 0.5, 1, 2, 3, 4, 5, 7, 10, 12, 24, 28, 32, 36, and 48 hours. The results were then quantitatively determined by high performance liquid chromatography (HPLC). Figure 10 shown.

[0113] Data Analysis:

[0114] like Figure 10 As shown, compared with the arsine precursor 2a, the half-life of the organic arsenic 5c in the blood is significantly prolonged, and its excretion through urine or feces is relatively slow.

[0115] The above experiments on microsomal enzymes confirmed that the benzene ring structure in organic arsenic inhibits the activity of microsomal enzyme 3A4, slows down metabolism in the body, prolongs the half-life in the body, and enhances the anti-tumor activity of organic arsenic.

[0116] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A novel targeted compound for treating various cancer types, characterized in that: The molecular structural formula of the targeting compound is: Wherein, the R1 is one of nitro and n-butoxy; R2 is one or more of alkyloxy groups.

2. A novel organoarsine targeting compound for treating various cancer types according to claim 1, characterized in that: The alkyl groups: methyl (—CH3), ethyl (—C2H5), n-propyl (—C3H7), isopropyl (—C3H7), n-butyl (—C4H9), isobutyl (—C4H9), tert-butyl (—C4H9), pentyl (—C5H 11 ) etc. until dodecyl (—C 12 H 25 ); Olefins : vinyl (—CH=CH2), propenyl (—CH2CH=CH2), butenyl (—C4H7), etc., including cis and trans isomers; alkoxy: methoxy (—OCH3), ethoxy (—OC2H5), n-propoxy (—OC3H7), isopropoxy (—OC3H7), n-butoxy (—OC4H9), etc. up to dodecyloxy (—OC 12 H 25 ); Alkylamino: methylamino (—NHCH3), ethylamino (—NHC2H5), dimethylamino (—N(CH3)2), diethylamino (—N(C2H5)2), etc.; Aminoalkyl: one or more of aminomethyl (—CH2NH2), 2-aminoethyl (—CH2CH2NH2), and 3-aminopropyl (—CH2CH2CH2NH2).

3. A novel organoarsine targeting compound for treating various cancer types according to claim 2, characterized in that: The arylalkylphenyl group includes tolyl, ethylphenyl, etc.; nitrophenyl includes p-nitrophenyl, o-nitrophenyl, m-nitrophenyl; hydroxyphenyl includes p-hydroxyphenyl, o-hydroxyphenyl, m-hydroxyphenyl; halogenated phenyl includes fluorophenyl, chlorophenyl, bromophenyl, iodophenyl (each halogen can be located at different positions of the benzene ring to form different isomers); one or more of naphthyl, anthracenyl, and biphenyl; The heterocyclic substituents are oxirane, oxirane, oxirane, acridinyl, pyrrolidinyl, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, furanyl, thiophene, piperidinyl, pyridyl, morpholinyl, piperazinyl, aza Oxy or thia One or more of; The halogen atom is one or more of fluorine, chlorine, bromine and iodine.

4. A method for preparing a novel organoarsine targeting compound for treating various types of cancer, for preparing the organoarsine targeting compound according to any one of claims 1 to 3, characterized in that: The preparation method comprises: Preparation of the arsenic precursors: 8-12 mL of 70% ammonium thioglycolate was heated to 50°C, and 3.5-4.5 g of phenylarsonic acid 1a or 1b was added. After 1 hour, 2 mL of 1,3-propanedithiol was slowly added dropwise via a constant pressure dropping funnel. After 2-4 hours of reaction, the resulting suspension was extracted with dichloromethane in small amounts, and the organic phases were combined and concentrated by rotary evaporation. The product was separated and purified on a silica gel column to obtain the arsenic precursors 2a and 2b. Synthesis of organic arsenic targeting compound: The triple-negative breast cancer targeting compound is prepared by mixing the arsenic precursors 2a and 2b with raw material compounds for reaction.

5. The method for preparing a novel organoarsine targeted compound for treating various cancer types according to claim 4, characterized in that: When the raw material compound is p-nitrophenylacetic acid 3a, the synthesis steps of the organic arsenic targeting compound are as follows: S101. Dissolve 4.5-5.5 mM p-nitrophenylacetic acid 3a in 20 mL of dichloromethane, add 7-8 mM oxalyl chloride and 30-50 μL of N,N-dimethylformamide, stir at room temperature for 40-50 min, and then dry the mixture to obtain p-nitrophenylacetyl chloride 3b as a pale yellow solid. S102. At room temperature, 4.5 mM p-nitrobenzeneacetyl chloride 3b was dissolved in 8-15 mL of dichloromethane, 3-4 mM of arsine precursors 2a and 2b were added, and pyridine was added as an acid-binding agent in an acyl chloride:triethylamine molar ratio of 1:1-1:

2. The products 3c and 3d were separated and purified by silica gel column chromatography. The preparation routes and structures of organoarsine targeting compounds 3c and 3d are as follows: Among them, in the organic arsenic targeting compound 3c, the position of 2-methyl-1,3,2-dithiarsine is 4; in the triple-negative breast cancer targeting compound 3d, the position of 2-methyl-1,3,2-dithiarsine is 2.

6. The claims of this patent cover a series of organic arsenic targeting compounds, specifically including compounds 4c, 4d, 5c, 5d, 6c, 6d and their derivatives.

7. Use of an organoarsine targeting compound prepared according to any one of claims 1 to 3 or the method for preparing an organoarsine targeting compound according to any one of claims 4 to 6 in treating various cancer types.

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