A semicarbazide derivative, its preparation method and application

Abstract

The present invention discloses a semicarbazide derivative and its preparation method and application, which belongs to the technical field of anti-lung cancer targeted drugs. The semicarbazide derivative of the present invention can inhibit the growth of H1299 cells in a dose-dependent manner, but has little toxicity to normal lung epithelial cells BEAS-2B. The semicarbazide derivative of the present invention can significantly inhibit the cloning of non-small cell lung cancer cells A549 and H1299, so it has a significant inhibitory effect on the proliferation of non-small cell lung cancer. The semicarbazide derivative provided by the present invention can inhibit the protein expression of GPX4 in non-small cell lung cancer A549 cells, and by inhibiting the protein expression of GPX4, promote the ferroptosis of non-small cell lung cancer cells, thereby playing a role in suppressing tumors.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-lung cancer targeted drugs, and in particular to a substituted semicarbazide derivative and a preparation method and application thereof. Background Art

[0002] Non-small cell lung cancer (NSCLC) is categorized into squamous cell carcinoma, adenocarcinoma, and large cell carcinoma. NSCLC accounts for approximately 85% of all lung cancer cases. The etiology of NSCLC is complex, with genetic mutations and genetic susceptibility playing a major role. Given its high morbidity and mortality, lung cancer remains a significant public health challenge, necessitating further research and improved prevention and treatment strategies.

[0003] Treatment options for non-small cell lung cancer include surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. Surgery is the preferred treatment for early-stage NSCLC and can achieve radical cure, but it is only suitable for localized tumors and patients whose physical condition allows it. Radiotherapy is often used for local tumor control and palliative treatment of advanced disease, although it may cause damage to normal tissue. Chemotherapy is effective for all stages of NSCLC, especially in the middle and late stages, but it has significant side effects and patient tolerance varies greatly. Immunotherapy has shown good results in some patients through PD-1 / PD-L1 inhibitors such as nivolumab and pembrolizumab, but it is only effective in some patients. Targeted therapy has shown significant efficacy in patients with gene mutations such as epidermal growth factor receptor (EGFR) and oncogenes such as ALK and ROS1. However, the problem of drug resistance to these drugs is serious, and the development of a new generation of targeted drugs is often needed to address it.

[0004] Drug resistance to targeted drugs includes target alterations due to gene mutations, activation of alternative signaling pathways, and increased drug excretion. In recent years, by synthesizing new molecular structures and optimizing existing drugs, researchers have continuously discovered and developed more effective therapeutics, such as the third-generation EGFR inhibitor osimertinib, which effectively overcomes the drug resistance of first- and second-generation EGFR inhibitors, providing NSCLC patients with more treatment options and hope. However, existing inhibitors not only have inhibitory effects on non-small cell lung cancer cells but are also cytotoxic to normal lung epithelial cells and lack specific targeting. Summary of the Invention

[0005] The present invention provides a semicarbazide derivative and a preparation method and application thereof. The semicarbazide derivative inhibits the protein expression of glutathione peroxidase 4, promotes the ferroptosis of non-small cell lung cancer cells, and thus plays a role in inhibiting tumors. The present invention provides a semicarbazide derivative with low reaction cost, high yield, simple and easy-to-control reaction process, and is suitable for industrial production.

[0006] The object of the present invention is to provide a semicarbazide derivative having the structural formula: Wherein, the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group.

[0007] As a preferred embodiment, the alkyl group is a C1-C5 alkyl group, the cycloalkyl group is cyclopropane, cyclobutane, cyclopentane or cyclohexane, and the substituted aromatic group is an alkyl-substituted phenyl group, an alkoxyphenyl group, a halogenated phenyl group or a phenylalkyl group.

[0008] As a preferred embodiment, the structural formula of the semicarbazide derivative is:

[0009]

[0010] A second object of the present invention is to provide a method for preparing the above-mentioned semicarbazide derivative, comprising the following steps:

[0011] Using 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I and tert-butoxybis(dimethylamino)methane of formula II as raw materials, toluene is added, the temperature is raised to 80-90°C with stirring, and the nucleophilic addition reaction is carried out for 8 hours. The product is filtered and dried to obtain (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-en-1-one of formula III.

[0012] (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propen-2-en-1-one of formula III is dissolved in glacial acetic acid, and ethyl acetoacetate and ammonium acetate are added. The temperature is raised to 110-125° C. with stirring, and the cyclization reaction is carried out for 36 hours. The extract is extracted to obtain an extract, which is dried, filtered, rotary evaporated, sanded, and purified to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV.

[0013] Ethanol and 80% hydrazine hydrate were added to the 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate ethyl ester, and the mixture was heated to 80° C. with stirring, refluxed for 48 hours, filtered and dried to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinic acid hydrazide of formula V.

[0014] The 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide and substituted isocyanate are used as raw materials, ethanol is added, the temperature is raised to 70-80°C while stirring, and the addition reaction is carried out for 8 hours. The mixture is filtered and dried to obtain a semicarbazide derivative. The synthesis route is as follows:

[0015] Wherein, the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group.

[0016] As a preferred embodiment, the molar ratio of 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I to tert-butoxybis(dimethylamino)methane of formula II is 1:1-3.

[0017] As a preferred embodiment, the molar ratio of (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propen-2-en-1-one of Formula III, ethyl acetoacetate and ammonium acetate is 1:1~2:6~9.

[0018] As a preferred embodiment, the molar ratio of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV to hydrazine hydrate is 1:20-30.

[0019] As a preferred embodiment, the molar ratio of 2-methyl-6-(quinolin-3-yl)nicotinamide hydrazide of formula V to isocyanate is 1:1.07-1.1.

[0020] As a preferred embodiment, during extraction, ethyl acetate and water are extracted three times in a volume ratio of 2:1, the extract is dried over anhydrous sodium sulfate, filtered, rotary evaporated, and sanded to obtain a crude product; during purification, the crude product is subjected to silica gel column chromatography using an eluent of ethyl acetate and petroleum ether in a volume ratio of 1:6.

[0021] The third object of the present invention is to provide a use of the above-mentioned semicarbazide derivative and a pharmaceutically acceptable salt thereof in the preparation of a protein expression inhibitor of glutathione peroxidase 4.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention provides a semicarbazide derivative and a preparation method and application thereof. The structural formula of the semicarbazide derivative is: Since ferroptosis is an iron-dependent form of cell death, it is characterized by the accumulation of lipid peroxidation and the destruction of cell membranes. Glutathione peroxidase 4 is a key regulatory factor in ferroptosis, which prevents the accumulation of lipid peroxidation by reducing phospholipid hydroperoxides, thereby protecting cells from the effects of ferroptosis. The semicarbazide derivatives of the present invention can inhibit the growth of H1299 cells in a dose-dependent manner, but have little toxicity to normal lung epithelial cells BEAS-2B. The semicarbazide derivatives of the present invention can significantly inhibit the cloning of non-small cell lung cancer cells A549 and H1299, so they have a significant inhibitory effect on the proliferation of non-small cell lung cancer. The semicarbazide derivatives provided by the present invention can inhibit the protein expression of GPX4 in non-small cell lung cancer A549 cells, and by inhibiting the protein expression of GPX4, promote the ferroptosis of non-small cell lung cancer cells, thereby playing a role in suppressing tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figures 1 and 2 These are the hydrogen and carbon spectra of (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-en-1-one of formula III of the present invention.

[0025] Figures 3 and 4 The hydrogen spectrum and carbon spectrum of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV of the present invention are shown.

[0026] Figures 5 and 6 The hydrogen spectrum and carbon spectrum of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide of formula V of the present invention are shown.

[0027] Figures 7 and 8 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 1 of the present invention.

[0028] Figures 9 and 10 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 2 of the present invention.

[0029] Figures 11 and 12 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 3 of the present invention.

[0030] Figures 13 and 14 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 4 of the present invention.

[0031] Figures 15 and 16 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 5 of the present invention.

[0032] Figures 17 and 18 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 6 of the present invention.

[0033] Figures 19 and 20 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 7 of the present invention.

[0034] Figures 21 and 22 These are the hydrogen and carbon spectra of the semicarbazide derivative prepared in Example 8 of the present invention.

[0035] Figure 23 The present invention uses the CCK8 method to test the in vitro activity of the compound. The compound is set at four concentrations of 5μM, 10μM, 20μM and 40μM. The treatment time is 48 hours, and the OD value is measured. Among them, Figure A is normal lung epithelial cells BEAS-2B, and Figure B is non-small cell lung cancer cells H1299.

[0036] Figure 24 The IC50 values of the active compounds were determined using the CCK8 method. The compounds were set at six concentrations: 1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM. The treatment time was 48 hours, and the OD values were measured to calculate the IC50 values.

[0037] Figure 25 This is a graph showing the inhibitory effect of the semicarbazide derivative prepared in Example 2 of the present invention on the proliferation of non-small cell lung cancer cells A549 and H1299.

[0038] Figure 26 This is a diagram showing the effect of the semicarbazide derivatives prepared in Example 2 of the present invention on inhibiting GPX4 expression in non-small cell lung cancer cells A549 and BEAS-2B. DETAILED DESCRIPTION

[0039] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention is further described below with reference to specific examples, but the examples are not intended to limit the present invention. The following experimental and detection methods are conventional methods unless otherwise specified; the reagents and raw materials are commercially available unless otherwise specified.

[0040] The present invention is directed to the problem mentioned in the background technology: the drug resistance problem of targeted drugs includes target changes caused by gene mutations, activation of alternative signaling pathways, and increased drug excretion. In recent years, by synthesizing new molecular structures and optimizing existing drugs, researchers have continuously discovered and developed more effective therapeutic drugs, such as the third-generation EGFR inhibitor osimertinib, which effectively overcomes the drug resistance problem of the first and second generation EGFR inhibitors, which provides more treatment options and hope for NSCLC patients. Therefore, studying new pathogenesis of non-small cell lung cancer and discovering new drug targets, and designing and synthesizing new targeted drugs are of great significance for the treatment of non-small cell lung cancer. Based on the above considerations, the present invention provides a semicarbazide derivative and its preparation method and application, and the technical solution of the present invention is further explained below.

[0041] The present invention provides a semicarbazide derivative, the structural formula of the semicarbazide derivative is: Wherein, the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group.

[0042] GPX4 (glutathione peroxidase 4) is a key regulator of ferroptosis. It reduces phospholipid hydroperoxides to prevent the accumulation of lipid peroxidation, thereby protecting cells from ferroptosis. The semicarbazide derivatives provided by the present invention inhibit GPX4 protein expression, promoting ferroptosis in non-small cell lung cancer cells, thereby suppressing tumors.

[0043] 1. Synthesis process of semicarbazide derivatives of the present invention

[0044] Synthesis of Intermediate 2 (Compound of Formula III): To a clean 25 mL round-bottom flask, add 0.5977 g (3.7317 mmol) of 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of Formula I and 0.8 mL (3.8742 mmol) of tert-butoxybis(dimethylamino)methane of Formula II, followed by 3.5 mL of toluene. The mixture was heated to 80°C with stirring and refluxed for 8 hours in an oil bath. TLC monitoring was performed until the reaction was complete. The reaction was stopped, cooled to room temperature, vacuum filtered, and dried at 80°C to obtain a yellow solid product, (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propen-2-en-1-one of Formula III, yielding 0.67 g in 83.5% yield.

[0045] Synthesis of intermediate 3 (compound of formula IV): In a clean 25 mL round-bottom flask, 0.5910 g (2.7475 mmol) of (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-en-1-one of formula III was added, followed by 15 mL of glacial acetic acid. After dissolution, 430 μL (3.3973 mmol) of ethyl acetoacetate and 1.95 g (0.0253 mol) of ammonium acetate were added in sequence. The mixture was heated to 125°C with stirring and refluxed. After 36 hours, TLC tracking monitoring was carried out until the reaction was completed, stirring was stopped, and the mixture was cooled to room temperature. 10 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate. The extract was dried over anhydrous sodium sulfate, filtered, rotary evaporated, and sanded to obtain a crude product. The crude product was subjected to silica gel column chromatography (eluent: A = V (ethyl acetate) / V (petroleum ether) = 1 / 6) to obtain a white solid product, namely, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV, 0.4003 g, and a yield of 51.06%.

[0046] Synthesis of Intermediate 4 (Compound of Formula V): In a clean 25 mL round-bottom flask, 0.4634 g (1.6484 mmol) of ethyl 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate, 10 mL of ethanol, and 2.3 mL of 80% hydrazine hydrate were added, respectively. The mixture was heated to 80° C. and refluxed with stirring. The mixture was kept in an oil bath for 48 h. The reaction was monitored by TLC until completion, after which the reaction was stopped. The mixture was cooled to room temperature, and a light yellow precipitate was formed. The product was vacuum filtered and dried to obtain a light yellow solid, i.e., 0.4115 g of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide of Formula V, in a yield of 93.48%.

[0047] The 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide and substituted isocyanate are used as raw materials, ethanol is added, the temperature is raised to 70-80°C with stirring, and the reaction is carried out for 8 hours. The mixture is filtered and dried to obtain a semicarbazide derivative. The synthetic route is as follows:

[0048] Wherein, the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group.

[0049] 2. The present invention synthesized the following 8 kinds of semicarbazide derivatives:

[0050] Example 1: The specific preparation steps of the semicarbazide derivative (YF-1) are:

[0051] In a 25 mL round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1 g, 0.3744 mmol) of formula V and 5 mL of anhydrous ethanol were added in sequence. After dissolving, 57 μL of 3,5-dimethylphenylisocyanate (0.0589 g, 0.4 mmol) was added. The mixture was stirred and heated to 80°C under reflux for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain the product having the structural formula: The yield of white solid product was 0.1106 g, with a yield of 71.32%.

[0052] Example 2: The specific preparation steps of the semicarbazide derivative (YF-2) are:

[0053] In a 25 mL round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1 g, 0.3744 mmol) of formula V and 5 mL of anhydrous ethanol were added in sequence. After dissolving, 45 μL of n-butyl isocyanate (0.0397 g, 0.4 mmol) was added. The mixture was stirred and heated to 80°C under reflux for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain the product having the structural formula: The yield of white solid product was 0.0907 g, with a yield of 66.16%.

[0054] Example 3: The specific preparation steps of the semicarbazide derivative (YF-3) are as follows:

[0055] In a 25 ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1 g, 0.3744 mmol) of formula V and 5 mL of anhydrous ethanol were added in sequence. After dissolving, 52 μL of p-methoxyphenyl isocyanate (0.0597 g, 0.4 mmol) was added. The mixture was stirred and heated to 80°C under reflux for 8 hours. The reaction was stopped after TLC detection. The mixture was filtered and washed with ethanol to obtain the product having the structural formula: The white solid product 4-(4-methoxyphenyl)-1-(2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinoyl)semicarbazide was obtained in an amount of 0.1292 g, with a yield of 82.92%.

[0056] Example 4: The specific preparation steps of the semicarbazide derivative (YF-4) are:

[0057] In a 25 ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1 g, 0.3744 mmol) of formula V and 5 mL of anhydrous ethanol were added in sequence. After dissolving, 49 μL of m-chlorophenyl isocyanate (0.0614 g, 0.4 mmmol) was added. The mixture was stirred and heated to 80°C under reflux for 8 hours. The reaction was stopped after TLC detection. The mixture was filtered and washed with ethanol to obtain the product having the structural formula: The yield of white solid product was 0.1096 g, with a yield of 69.68%.

[0058] Example 5: The specific preparation steps of the semicarbazide derivative (YF-5) are:

[0059] In a 25ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1g, 0.3744mmol) of formula V and 5mL of anhydrous ethanol were added in sequence. After dissolving, 45μL of cyclopentyl isocyanate (0.044g, 0.4mmmol) was added. The mixture was stirred and heated to 80℃ and refluxed for 8 hours. The reaction was stopped after TLC detection. Filtered and washed with ethanol to obtain the structural formula The yield of white solid product was 0.12 g, with a yield of 65.6%.

[0060] Example 6: The specific preparation steps of the semicarbazide derivative (YF-6) are:

[0061] In a 25ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1g, 0.3744mmol) of formula V and 5mL of anhydrous ethanol were added in sequence. After dissolving, 54μL of p-toluene isocyanate (0.53g, 0.4mmmol) was added. The mixture was stirred and heated to 80℃ under reflux. The reaction was allowed to proceed for 8 hours. The reaction was stopped after TLC detection. Filtered and washed with ethanol to obtain the product with the structural formula The yield of the white solid product was 0.10 g, with a yield of 66.7%.

[0062] Example 7: The specific preparation steps of the semicarbazide derivative (YF-7) are:

[0063] In a 25ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1g, 0.3744mmol) of formula V and 5mL of anhydrous ethanol were added in sequence. After dissolving, 51μL of m-toluene isocyanate (0.053g, 0.4mmmol) was added. The mixture was stirred and heated to 80℃ under reflux for 8 hours. The reaction was stopped after TLC detection. Filtered and washed with ethanol to obtain the structural formula The yield of white solid product was 0.11 g, with a yield of 73.3%.

[0064] Example 8: The specific preparation steps of semicarbazide derivative (YF-8) are as follows:

[0065] In a 25ml round-bottom flask, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide (0.1g, 0.3744mmol) of formula V and 5mL of anhydrous ethanol were added in sequence. After dissolving, 49μL of benzyl isocyanate (0.053g, 0.4mmmol) was added. The mixture was stirred and heated to 80℃ and refluxed for 8 hours. The reaction was stopped after TLC detection. Filtered and washed with ethanol to obtain the structural formula The yield of the white solid product was 0.10 g, with a yield of 66.7%.

[0066] III. The physicochemical parameters and structural parameters of the semicarbazide derivatives of the structural formulas of Examples 1 to 8 are shown in Table 1 below.

[0067] Table 1 Physicochemical parameters and structural parameters of intermediates and 8 kinds of semicarbazide derivatives

[0068]

[0069]

[0070]

[0071]

[0072]

[0073] IV. Performance testing of the semicarbazide derivatives prepared by the present invention in terms of application

[0074] First, a preliminary in vitro toxicity test was conducted on the semicarbazide derivatives (YF-1, YF-2, YF-3, and YF-4) prepared by the present invention using the CCK8 method. The semicarbazide derivatives were set at four concentrations of 5 μM, 10 μM, 20 μM, and 40 μM, and treated for 48 hours. The OD values were measured to screen out active compounds with concentration-dependent and better effects. Figure 23 As shown, the product YF-2 can inhibit the growth of H1299 cells in a dose-dependent manner, but has little toxicity to normal lung epithelial cells BEAS-2B. Therefore, YF-2 was used as the active compound for the next experiment.

[0075] The IC50 value of the active compound YF-2 was further tested using A549, H1299, H1975, and BEAS-2B cells, with the semicarbazide derivative at five concentrations of 1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM for 48 hours. Figure 24 The results showed that the IC50 values of YF-2 for the four cell lines were: A549: 10.41±0.72μM, H1299: 19.42±0.23μM; H1975: 23.74±0.55μM, BEAS-2B: 162.2±0.18μM.

[0076] Since YF-2 is more sensitive to A549 and H1299 cells, we further tested the inhibitory effect of YF-2 on the clone formation ability of non-small cell lung cancer using these two cells. We treated A549 and H1299 cells with 20μM YF-2 and an equal dose of DMSO, respectively. After 7 days, the samples were harvested and stained with 0.1% crystal violet solution for 10 minutes. After drying, the samples were photographed. Figure 25 As shown, the results showed that YF-2 could significantly inhibit the clone formation of the two cell lines, indicating that YF-2 has a significant inhibitory effect on the proliferation of non-small cell lung cancer.

[0077] GPX4 protein plays a key role in inhibiting ferroptosis during the development and progression of non-small cell lung cancer. Designing compounds that can regulate GPX4 expression to induce tumor cell death has important therapeutic potential and clinical application value. We further treated A549 and BEAS-2B cells with DMSO, YF-2 (10μM) and YF-2 (20μM). After 24 hours, samples were collected and Western Blot was used to detect the expression of GPX4 protein. GAPDH was used as an internal control. Figure 26 As shown in the results, YF-2 can inhibit the protein expression of GPX4 in A549 cells, but does not affect the expression of GPX4 in BEAS-2B cells, which are insensitive to YF-2. This suggests that YF-2 may promote ferroptosis in non-small cell lung cancer cells by inhibiting GPX4 expression, thereby achieving a tumor-suppressing effect.

[0078] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A semicarbazide derivative, characterized in that The structural formula of the semicarbazide derivative is: , Wherein, the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group; the substituted aromatic group is an alkyl-substituted phenyl group, an alkoxyphenyl group, a halogenated phenyl group or a benzyl group.

2. The semicarbazide derivative according to claim 1, characterized in that The alkyl group is a C1~C5 alkyl group, and the cycloalkyl group is cyclopropane, cyclobutane, cyclopentane or cyclohexane.

3. The semicarbazide derivative according to claim 1, characterized in that The structural formula of the semicarbazide derivative is: , , , , , , or .

4. A method for preparing the semicarbazide derivative according to claim 1, characterized in that: The following steps are involved: Using 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I and tert-butoxybis(dimethylamino)methane of formula II as raw materials, toluene is added, and the temperature is raised to 80-90°C while stirring to allow a nucleophilic addition reaction to occur, thereby obtaining (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propen-2-en-1-one of formula III; The (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propen-2-en-1-one is dissolved in glacial acetic acid, and ethyl acetoacetate and ammonium acetate are added. The temperature is raised to 110-125° C. while stirring to allow a cyclization reaction to occur. Extraction is performed to obtain an extract, which is dried, filtered, rotary evaporated, sanded, and purified to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV; Adding ethanol and hydrazine hydrate to the 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinic acid ethyl ester, heating to 80° C. with stirring, and performing reflux reaction to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinic acid hydrazide of formula V; Using the aforementioned 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinamide hydrazide and isocyanate as raw materials, ethanol is added, and the mixture is heated to 70-80°C while stirring to allow an addition reaction to occur. The mixture is then filtered and dried to obtain a semicarbazide derivative. The synthetic route is as follows: , wherein the R group is an alkyl group, a cycloalkyl group or a substituted aromatic group.

5. The preparation method according to claim 4, characterized in that The molar ratio of 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I to tert-butoxybis(dimethylamino)methane of formula II is 1:1-3.

6. The preparation method according to claim 4, characterized in that The molar ratio of (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-en-1-one of formula III, ethyl acetoacetate and ammonium acetate is 1:1-2:6-9.

7. The preparation method according to claim 4, characterized in that The molar ratio of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV to hydrazine hydrate is 1:20-30.

8. The preparation method according to claim 4, characterized in that The molar ratio of 2-methyl-6-(quinolin-3-yl)nicotinylhydrazide of formula V to isocyanate is 1:1.07-1.

1.

9. The preparation method according to claim 4, characterized in that During extraction, ethyl acetate and water in a volume ratio of 2:1 are used for extraction. The extract is dried over anhydrous sodium sulfate, filtered, rotary evaporated, and sand made to obtain a crude product. During purification, the crude product is subjected to silica gel column chromatography using an eluent of ethyl acetate and petroleum ether in a volume ratio of 1:

6.

10. Use of the semicarbazide derivative according to any one of claims 1 to 3 and a pharmaceutically acceptable salt thereof in the preparation of a glutathione peroxidase 4 protein expression inhibitor.

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