Semicarbazide derivative as well as preparation method and application thereof

Abstract

The invention discloses a semicarbazide derivative as well as a preparation method and application thereof, and belongs to the technical field of anti-lung cancer targeted drugs, the semicarbazide derivative can inhibit the growth of H1299 cells in a dose-dependent manner, but has very small toxicity to normal pulmonary epithelial cells BEAS-2B. The semicarbazide derivative disclosed by the invention can be used for remarkably inhibiting the clone formation of non-small cell lung cancer cells A549 and H1299, so that the semicarbazide derivative has a remarkable inhibiting effect on the proliferation of the non-small cell lung cancer. The semicarbazide derivative provided by the invention can inhibit the protein expression of GPX4 in non-small cell lung cancer A549 cells, and promotes ferroptosis of the non-small cell lung cancer cells by inhibiting the protein expression of GPX4, thereby playing a role in inhibiting 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 is divided into squamous cell carcinoma, adenocarcinoma and large cell carcinoma. Non-small cell lung cancer accounts for about 85% of all lung cancer cases. The causes of non-small cell lung cancer are complex, and gene mutations and genetic susceptibility are a major cause of non-small cell lung cancer. Given its high morbidity and mortality, lung cancer remains a major challenge to public health, and further research and improved prevention and treatment strategies are needed.

[0003] Treatments for non-small cell lung cancer include surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. Surgery is the first choice for early-stage NSCLC and can achieve radical treatment, but it is only suitable for localized tumors and patients with physical conditions that allow 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 the side effects are obvious 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 for some patients. Targeted therapy has shown significant efficacy in patients with gene mutations such as epidermal growth factor receptor (EGFR) and proto-oncogenes such as ALK and ROS1. However, the problem of drug resistance to these drugs is serious, and a new generation of targeted drugs often needs to be developed to deal with it.

[0004] The drug resistance 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, providing more treatment options and hope for NSCLC patients. 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 do not have specific targeting. Summary of the invention

[0005] The present invention provides a semicarbazide derivative and a preparation method and application thereof. The semicarbazide derivative is used to inhibit the protein expression of glutathione peroxidase 4, promote the ferroptosis of non-small cell lung cancer cells, and thus play 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 suitable for industrial production.

[0006] The object of the present invention is to provide 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.

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

[0011] Take 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I and tert-butoxybis(dimethylamino)methane of formula II as raw materials, add toluene, heat to 80-90°C under stirring, carry out nucleophilic addition reaction for 8h, filter and dry to obtain (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-ene-1-one of formula III.

[0012] Use glacial acetic acid to dissolve (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-ene-1-one of formula III, then add ethyl acetoacetate and ammonium acetate, heat to 110-125° C. in a stirring state, carry out cyclization reaction for 36 hours, extract to obtain an extract, dry, filter, rotary evaporate, sand make, and purify to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV.

[0013] Add ethanol and hydrazine hydrate with a mass concentration of 80% to the 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinic acid ethyl ester, heat to 80° C. with stirring, reflux for 48 hours, filter and dry 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)nicotinylhydrazide and substituted isocyanate are used as raw materials, ethanol is added, the temperature is raised to 70-80° C. under stirring, the addition reaction is carried out for 8 hours, and the mixture is filtered and dried to obtain a semicarbazide derivative. The synthesis route thereof 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)propene-2-en-1-one of formula III, ethyl acetoacetate and ammonium acetate is 1:1 to 2:6 to 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 the 2-methyl-6-(quinolin-3-yl)nicotinylhydrazide of formula V to the isocyanate is 1:1.07-1.1.

[0020] As a preferred embodiment, during extraction, ethyl acetate and water are used in a volume ratio of 2:1 for extraction three times, 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, characterized by the accumulation of lipid peroxidation and the destruction of cell membranes, glutathione peroxidase 4 is a key regulator of ferroptosis, which prevents the accumulation of lipid peroxidation by reducing phospholipid hydroperoxides, thereby protecting cells from the effects of ferroptosis. The aminourea 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 aminourea 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 aminourea derivatives provided by the present invention can inhibit the protein expression of GPX4 in non-small cell lung cancer A549 cells, and promote the ferroptosis of non-small cell lung cancer cells by inhibiting the protein expression of GPX4, thereby playing a role in inhibiting tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1-2 The hydrogen spectrum and carbon spectrum 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 are shown.

[0025] Figure 3-4 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] Figure 5-6 The hydrogen spectrum and carbon spectrum of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinylhydrazide of formula V of the present invention are shown.

[0027] Figure 7-8 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 1 of the present invention.

[0028] Figure 9-10 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 2 of the present invention.

[0029] Figure 11-12 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 3 of the present invention.

[0030] Figure 13-14 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 4 of the present invention.

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

[0032] Figure 17-18 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 6 of the present invention.

[0033] Figure 19-20 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 7 of the present invention.

[0034] Figure 21-22 The hydrogen spectrum and carbon spectrum of the semicarbazide derivative prepared in Example 8 of the present invention.

[0035] Fig.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 5 μM, 10 μM, 20 μM and 40 μM. The treatment time is 48 hours, and the OD value is measured; wherein, Figure A is a normal lung epithelial cell BEAS-2B, and Figure B is a non-small cell lung cancer cell H1299.

[0036] Fig.24 The IC50 value of the active compound was determined by CCK8 method in the present invention. The compound was set at six concentrations of 1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM, and the treatment time was 48 hours. The OD value was measured and the IC50 value was calculated.

[0037] Fig.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] Fig.26 This is a diagram showing the effect of the semicarbazide derivative prepared in Example 2 of the present invention on inhibiting the expression of GPX4 in non-small cell lung cancer cells A549 and BEAS-2B. DETAILED DESCRIPTION

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

[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, 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 a preparation method and application thereof. The technical scheme 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] Since GPX4 (glutathione peroxidase 4) is a key regulator of ferroptosis, it prevents the accumulation of lipid peroxidation by reducing phospholipid hydroperoxides, thereby protecting cells from ferroptosis. The semicarbazide derivatives provided by the present invention inhibit the protein expression of GPX4 and promote the ferroptosis of non-small cell lung cancer cells, thereby inhibiting tumors.

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

[0044] Synthesis of intermediate 2 (compound of formula III): Take a clean 25mL round-bottom flask, add 0.5977g (3.7317mmol) of 1-(1H-pyrrolo[2,3-b]pyridin-5-yl)ethanone of formula I, 0.8mL (3.8742mmol) of tert-butoxybis(dimethylamino)methane of formula II, and 3.5mL toluene, heat to 80℃ and reflux under stirring, keep in oil bath for 8h, and monitor by TLC until the reaction is completed. Stop the reaction and cool to room temperature, vacuum filter, and dry at 80℃ to obtain a yellow solid product, that is, (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-ene-1-one of formula III, 0.67g, with a yield of 83.5%.

[0045] Synthesis of intermediate 3 (compound of formula IV): In a clean 25 mL round-bottom flask, add 0.5910 g (2.7475 mmol) of (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propylene-2-ene-1-one of formula III, then add 15 mL of glacial acetic acid, and after dissolving, add 430 μL (3.3973 mmol) of ethyl acetoacetate and 1.95 g (0.0253 mol) of ammonium acetate in sequence, and heat to 125 °C under stirring for reflux reaction. 36h, TLC tracking monitoring until the reaction is completed, then stirring is stopped, cooled to room temperature, 10mL of water is added, and then extracted with 20mL of ethyl acetate for 3 times. The extract is dried over anhydrous sodium sulfate, filtered, rotary evaporated, and sand made to obtain a crude product. The crude product is chromatographed on a silica gel column (eluent: A=V(ethyl acetate) / V(petroleum ether)=1 / 6) to obtain a white solid product, that is, 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinate of formula IV, 0.4003g, yield 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 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, and the mixture was heated to 80° C. and refluxed under stirring. The mixture was kept in an oil bath for 48 h. The reaction was stopped after TLC tracking and monitoring until the reaction was completed. The mixture was cooled to room temperature and a light yellow precipitate was precipitated. The light yellow solid product was obtained by vacuum filtration and drying, i.e., 0.4115 g of 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinylhydrazide of formula V was obtained, with a yield of 93.48%.

[0047] The 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinylhydrazide and substituted isocyanate are used as raw materials, ethanol is added, the temperature is raised to 70-80° C. under stirring for 8 hours, and then 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 and refluxed for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain a product having the structural formula: The 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 and refluxed for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain a 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:

[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-methoxyphenylisocyanate (0.0597 g, 0.4 mmol) was added. The mixture was stirred and heated to 80°C and refluxed for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain a 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 and refluxed for 8 hours. The reaction was stopped by TLC detection. The mixture was filtered and washed with ethanol to obtain a 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 25 ml round-bottom flask, add 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 in sequence, add 45 μL of cyclopentyl isocyanate (0.044 g, 0.4 mmmol) after dissolving, stir and heat to 80 °C and reflux, react for 8 hours, TLC detection shows that the reaction is complete, and the reaction is stopped. Filter and wash 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 25 ml round-bottom flask, add 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 in sequence, add 54 μL of p-toluene isocyanate (0.53 g, 0.4 mmmol) after dissolving, stir and heat to 80 ° C and reflux, react for 8 hours, TLC detection shows that the reaction is complete, and the reaction is stopped. Filter and wash with ethanol to obtain the structural formula 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 25 ml round-bottom flask, add 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 in sequence, add 51 μL of m-toluene isocyanate (0.053 g, 0.4 mmmol) after dissolving, stir and heat to 80 °C under reflux, react for 8 hours, TLC detection shows that the reaction is complete, and the reaction is stopped. Filter and wash 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 the semicarbazide derivative (YF-8) are:

[0065] In a 25 ml round-bottom flask, add 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 in sequence, add 49 μL of benzyl isocyanate (0.053 g, 0.4 mmmol) after dissolving, stir and heat to 80 °C and reflux, react for 8 hours, TLC detection shows that the reaction is complete, and the reaction is stopped. Filter and wash with ethanol to obtain the structural formula 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 application

[0074] First, the CCK8 method was used to conduct a preliminary in vitro toxicity test on the semicarbazide derivatives (YF-1, YF-2, YF-3 and YF-4) prepared by the present invention. The semicarbazide derivatives were set at 4 concentrations of 5 μM, 10 μM, 20 μM and 40 μM, treated for 48 hours, and their OD values ​​were measured to screen out active compounds with concentration-dependency and good effects. Fig.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 an active compound for the next experiment.

[0075] The IC50 value of the active compound YF-2 was further tested. Four cell lines, A549, H1299, H1975 and BEAS-2B, were used, and the semicarbazide derivatives were set at 5 concentrations of 1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM and 50 μM for 48 hours. Fig.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 used these two cells to further test the inhibitory effect of YF-2 on the clone formation ability of non-small cell lung cancer. We treated A549 and H1299 cells with 20μM YF-2 and an equal dose of DMSO, respectively. After 7 days, the samples were collected and stained with 0.1% crystal violet stain for 10 minutes. After drying, the samples were photographed. Fig.25 As shown, the results showed that YF-2 could significantly inhibit the clone formation of both cells, 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 in the development 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 (10 μM) and YF-2 (20 μM), collected samples after 24 hours and detected the expression of GPX4 protein by Western Blot, GAPDH as an internal reference, such as Fig.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 that are insensitive to YF-2. This suggests that YF-2 may promote ferroptosis of non-small cell lung cancer cells by inhibiting GPX4 expression, thereby achieving the effect of tumor suppression.

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

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.

2. The substituted semicarbazide derivative according to claim 1, characterized in that 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.

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

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, adding toluene, heating to 80-90° C. under stirring, a nucleophilic addition reaction occurs, and (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-ene-1-one of formula III is obtained; The (E)-3-(dimethylamino)-1-(1H-pyrrolo[2,3-b]pyridin-3-yl)propene-2-en-1-one is dissolved in glacial acetic acid, and ethyl acetoacetate and ammonium acetate are added, and the temperature is raised to 110-125° C. under stirring to cause a cyclization reaction, and 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; Add ethanol and hydrazine hydrate to the 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinic acid ethyl ester, raise the temperature to 80° C. under stirring, and perform reflux reaction to obtain 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinylhydrazide of formula V; The 2-methyl-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)nicotinylhydrazide and isocyanate are used as raw materials, ethanol is added, the temperature is raised to 70-80° C. under stirring, an addition reaction occurs, and the mixture is filtered and dried to obtain a semicarbazide derivative. The synthesis 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]pyridine-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 the 2-methyl-6-(quinolin-3-yl)nicotinylhydrazide of formula V to the 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 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.

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 protein expression inhibitor of glutathione peroxidase 4.

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