2-phenyl methylene hydrazine-1-sulfamamide derivative compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a 2-phenyl methylene hydrazine-1-amino sulfamide derivative as well as a preparation method and application of the 2-phenyl methylene hydrazine-1-amino sulfamide derivative. The antitumor cell proliferation activity of the compound is evaluated through an MTT method cancer cell proliferation inhibition experiment, and experimental data shows that the compounds with the structures as shown in the formulas I and II have definite and high antitumor cell proliferation activity, and can be used for preparing antitumor drugs. The conclusion proves that the 2-phenyl methylene hydrazine-1-sulfanilamide derivatives with the structures shown in the formula I and the formula II can be used for preparing the anti-tumor drugs and have application prospects in the anti-tumor aspect.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of 2-phenyl methylene hydrazine-1-amino sulfamide derivatives and its preparation method and use. BACKGROUND

[0002] Malignant tumor endangers human life and health, has been widely concerned in the world. Finding effective, good activity antitumor drug is an important task of treating tumor. At present, the main means of treating cancer is radiotherapy and chemotherapy, but the overall survival rate is low. Breast cancer, ovarian cancer and cervical cancer are three major cancers that endanger women's life and health, and the incidence of cervical cancer is high. There is no safe and effective drug in clinic. Therefore, it is urgent to explore new effective treatment drugs for cervical cancer to improve its effectiveness in clinical treatment. SUMMARY

[0003] The technical problem to be solved by the present application is to overcome the defects and deficiencies of the relative lack of existing cervical cancer treatment drugs. The present application provides a kind of 2-phenyl methylene hydrazine-1-amino sulfamide derivatives and its preparation method and use.

[0004] In order to solve the above technical problems, first of all, the present application adopts the following technical scheme: The present application provides a kind of 2-phenyl methylene hydrazine-1-amino sulfamide derivatives, characterized in that the compound and its pharmaceutically acceptable salt have the structure shown in formula I and formula II:

[0005] Secondly, the present application also provides the application of the pharmaceutical composition containing the above-mentioned compound in treating or inhibiting the proliferation of malignant tumor cells.

[0006] Thirdly, the present application provides a preparation method of the above-mentioned compound, comprising the following steps: S1: nucleophilic substitution reaction of compound 1 2H-indazole-7-methyl formate and compound 2 4-fluorobenzaldehyde under alkaline conditions to generate compound 3 methyl 2-(4-formylphenyl)-2H-indole-7-carboxylate; S2: condensation reaction of aminothiourea, 2-methyl aminothiourea and compound 3 methyl 2-(4-formylphenyl)-2H-indole-7-carboxylate under acidic conditions to generate compound I and compound II.

[0007] The chemical reaction formula of the 2-phenyl methylene hydrazine-1-amino sulfamide derivative provided by the present application is as follows:

[0008] The reaction solvent of the nucleophilic substitution reaction in step S1 is selected from aprotic solvents such as DMF, DMAC, DMSO, and the like, with DMF and DMAC being preferred; the base is selected from cesium carbonate, sodium carbonate, potassium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium amide, potassium amide, potassium hydroxide, sodium hydride, and the like, with potassium carbonate being preferred; the reaction temperature of the nucleophilic substitution is 100-150°C, with 150°C being preferred.

[0009] The reaction solvent of the condensation reaction in step S2 is a polar protic solvent selected from methanol, ethanol, isopropanol, and the like, with methanol being preferred; the acid catalyst used is selected from formic acid, acetic acid, hydrochloric acid, and the like, with acetic acid being preferred. The condensation reaction temperature is 50-80°C, with 80°C being preferred.

[0010] The present application provides 2-phenylmethylenehydrazine-1-amino sulfamide derivatives, a preparation method and use thereof. The 2-phenylmethylenehydrazine-1-amino sulfamide derivatives or a pharmaceutically acceptable salt thereof are compounds of formula I and formula II, which have excellent performance in the application of resisting malignant tumor cell proliferation.

[0011] The benzopyrazole derivatives of formula I and formula II provided by the present application are a new series of anti-tumor proliferation compounds. In terms of structural characteristics, the benzopyrazole ring is the basic core structure, and after introducing specific substituent groups into the basic core structure, the compounds have excellent therapeutic effect in treating and / or preventing various malignant tumor proliferations of animals (including humans). The compounds of formula I and formula II have outstanding inhibitory effect in human ovarian cancer cell (SKOV3) anti-proliferation experiments, and can effectively resist the proliferation of SKOV3 cells. The conclusion proves that the compounds of formula I and formula II have the potential to be developed as new cancer treatment drugs. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 NMR H spectrum of formula I Figure 2 NMR C spectrum of formula I Figure 3 Mass spectrum of formula I Figure 4 NMR H spectrum of formula II Figure 5 NMR C spectrum of formula II Figure 6 Mass spectrum of formula II DETAILED DESCRIPTION

[0013] This invention discloses 2-phenylmethylenehydrazine-1-aminothioamide derivatives, their preparation methods, and uses. Those skilled in the art can refer to this document and appropriately modify the process parameters to achieve the desired results. It is particularly important to note that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included within this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0014] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below with reference to specific embodiments.

[0015] All the raw material compounds described in this invention can be purchased commercially. The compounds shown in Formula I and Formula II can be synthesized using conventional compound synthesis methods according to the following synthetic routes. Example 1

[0016] Compound 1 (3.46 mmol, 1.0 eq), compound 2 (5.19 mmol, 1.5 eq), potassium carbonate (10.38 mmol, 3 eq), and DMF were added to the reaction flask. After reacting at 150 °C for about 3 hours, the reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the reaction solution was extracted three times with ethyl acetate and saturated brine. The organic phase was back-extracted, washed, dried, concentrated, and separated by column chromatography to obtain intermediate 3 with a yield of about 84%.

[0017] Compound 3 (1.07 mmol, 1 eq), aminothiourea (1.39 mmol, 1.3 eq), and 2-methylaminothiourea (1.39 mmol, 1.3 eq) were added to the mixture. The pH was adjusted to 6 with acetic acid, and the mixture was reacted at 80 °C for 4 h. After the reaction was completed, the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (V (EA): V / (PE) = 1:3) to obtain the target products I and II in 85% yield. Example 2

[0018] Compound 1 (3.46 mmol, 1.0 eq), compound 2 (5.19 mmol, 1.5 eq), sodium carbonate (10.38 mmol, 3 eq), and DMF were added to the reaction flask. The reaction was carried out at 135 °C for about 4.5 hours. The reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the reaction solution was extracted three times with ethyl acetate and saturated brine. The organic phase was back-extracted, washed, dried, concentrated, and separated by column chromatography to obtain intermediate 3 with a yield of about 78%.

[0019] Compound 3 (1.07 mmol, 1 eq), aminothiourea (1.39 mmol, 1.3 eq), and 2-methylaminothiourea (1.39 mmol, 1.3 eq) were added to the mixture, and the pH was adjusted to 6 with hydrochloric acid. The mixture was reacted at 70 °C for 5 h. After the reaction was completed, the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (V (EA): V / (PE) = 1:3) to obtain the target products I and II in 80% yield. Example 3

[0020] Compound 1 (3.46 mmol, 1.0 eq), compound 2 (5.19 mmol, 1.5 eq), sodium hydroxide (10.38 mmol, 3 eq), and DMF were added to the reaction flask. After reacting at 125 °C for about 5 hours, the reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the reaction solution was extracted three times with ethyl acetate and saturated brine. The organic phase was back-extracted, washed, dried, concentrated, and separated by column chromatography to obtain intermediate 3 with a yield of about 75%.

[0021] Compound 3 (1.07 mmol, 1 eq), aminothiourea (1.39 mmol, 1.3 eq), and 2-methylaminothiourea (1.39 mmol, 1.3 eq) were added to the mixture. The pH was adjusted to 5 with acetic acid, and the mixture was reacted at 60 °C for 4 h. After the reaction was completed, the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (V (EA): V / (PE) = 1:3) to obtain the target products I and II in 70% yield. Example 4

[0022] Compound 1 (3.46 mmol, 1.0 eq), compound 2 (5.19 mmol, 1.5 eq), potassium carbonate (10.38 mmol, 3 eq), and DMAC were added to the reaction flask. After reacting at 120 °C for about 6 hours, the reaction progress was monitored by thin-layer chromatography. After the reaction was completed, the reaction solution was extracted three times with ethyl acetate and saturated brine. The organic phase was back-extracted, washed, dried, concentrated, and separated by column chromatography to obtain intermediate 3 with a yield of about 70%.

[0023] Compound 3 (1.07 mmol, 1 eq), aminothiourea (1.39 mmol, 1.3 eq), and 2-methylaminothiourea (1.39 mmol, 1.3 eq) were added to the mixture. The pH was adjusted to 5 with acetic acid, and the mixture was reacted at 50 °C for 7 h. After the reaction was completed, the filtrate was evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography (V (EA): V / (PE) = 1:3) to obtain the target products I and II in 69% yield.

[0024] The target compounds prepared in Examples 1, 2, 3, and 4 of this invention have structures as shown in Formulas I and II. The compounds were analyzed by 1H NMR and mass spectrometry data, and the data analysis is shown in Table 1.

[0025] Table 1. 1H NMR, 1C NMR, and mass spectrometry data of the compounds shown in Formulas I and II.

[0026] Example 5 Cytotoxicity tests and quantitative toxicity tests were performed on the Formula I and Formula II compounds prepared in Examples 1, 2, 3, and 4, as well as other control compounds: 5.1 Preparation of positive control and compound solutions Weigh 3.0 mg of the positive control niraparib, dissolve it in DMSO to prepare a stock solution of 20 mg / mL, and store it at -20℃ for later use. Prepare the target compound solution in the same way, and dilute the stock solution of the test compound to the required concentration with 90% DMEM high glucose medium before use.

[0027] 5.2 Preparation of PBS and MTT solutions Phosphate-buffered saline (PBS) is prepared by dissolving PBS powder in ultrapure water, adjusting the pH to 7-8, sterilizing, filtering through a 0.22 μm microporous membrane, and storing at 4°C. MTT solution is prepared by weighing 0.25 g of MTT, dissolving it in 50 mL of pre-prepared PBS solution, stirring for half an hour in the dark to achieve a concentration of 5 mg / mL, filtering through a 0.22 μm microporous membrane, and storing at -20°C in the dark. It is crucial that this solution be prepared fresh and used immediately, and that the preparation process be conducted in the dark.

[0028] 5.3 MTT assay for inhibiting cancer cell proliferation Preparation of phosphate-buffered saline (PBS): Dissolve commercially available PBS powder in ultrapure water, adjust the pH to approximately 7.4, sterilize at high temperature, filter through a 0.22 μm microporous membrane for sterilization, and store the resulting solution at 4°C for later use.

[0029] Preparation of MTT solution: Accurately weigh 0.25 g of MTT powder and dissolve it in 50 mL of pre-prepared and sterilized PBS solution. Stir magnetically for 30 minutes in the dark until completely dissolved, resulting in a solution concentration of 5 mg / mL. After filtration through a 0.22 μm microporous membrane, aliquot and store at -20°C in the dark.

[0030] Log-phase ovarian cancer cells (SKOV-3) were harvested, digested with trypsin, resuspended, and the cell concentration was adjusted to 5 × 10⁻⁶ cells / mL.4 Cells were seeded at 100 μL / well in 96-well plates and allowed to adhere for 24 h. The culture medium was then discarded, and different concentrations of the compound (5, 10, 20 μg / mL) were added. Each concentration was used in triplicate, with corresponding concentrations of PBS as a control, a cell-free nulling group, and a positive control group. Cells were cultured at 37°C and 5% CO2 for 48 h, followed by incubation at 37°C and 5% CO2 for 4 h with 20 μL of 5 mg / mL MTT. The supernatant was discarded, and 150 μL of DMSO was added to each well. The cells were shaken for 10 minutes, and the OD value was measured at 490 nm using a microplate reader. The cell inhibition rate and half-maximal inhibitory concentration (IC50) were calculated. 50 ).

[0031] The data are shown in Table 2, which lists the evaluation results of the effects of Formula I and Formula II and other control compounds on the proliferation activity of SKOV-3 cells.

[0032] Table 2. Effects of compounds on SKOV-3 cell viability

[0033] As shown in Table 2, in the MTT assay for inhibiting cancer cell proliferation, Formulas I and II, administered at a dose of 20 μg / mL, exhibited certain anti-proliferative properties against cancer cells. The results showed that, compared to the positive control niraparib, Formulas I and II significantly inhibited the proliferation of SKOV-3 cervical cancer cells, demonstrating a clear and prominent inhibitory effect. These results indicate that for the core structure of 2-phenylmethylenehydrazine-1-aminothioamide derivatives, only specific groups introduced onto the N atom of the tail-end aminothiourea exhibit good anti-cancer cell proliferation properties, and the structural type of the group has no obvious influence on the overall anti-proliferative activity of the molecule. Therefore, it can be concluded that the type of group has an unpredictable effect on the overall anti-cancer cell proliferation activity of the molecule.

[0034] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A 2-phenylmethylenehydrazine-1-aminothioamide derivative, characterized in that, The compounds and their pharmaceutically usable salts have the structures shown in Formula I and Formula II:

2. A pharmaceutical composition, characterized in that, The active ingredient is the compound of claim 1, and it also contains one or more pharmaceutical carriers or one or more known pharmaceutical diluents.

3. The use of the compound according to claim 1 or the pharmaceutical composition according to claim 2 in an antitumor drug.

4. A method for preparing the compound according to claim 1, characterized in that, Includes the following steps: Step a): Compound 1 and compound 2 undergo a nucleophilic substitution reaction under basic conditions to generate compound 3. Step b) Thiourea, 2-methylthiourea, and compound 3 undergo a condensation reaction under acidic conditions to produce compounds shown in formulas I and II:

5. In the preparation method shown in step a) of claim 4, the reaction solvent for the nucleophilic substitution reaction is selected from aprotic solvents such as DMF, DMAC, and DMSO; the base is selected from cesium carbonate, sodium carbonate, potassium carbonate, sodium tert-butoxide, potassium tert-butoxide, sodium amino, potassium amino, potassium hydroxide, sodium hydride, etc.; and the reaction temperature is 100-150℃.

6. The reaction solvent for step b) condensation reaction is a polar protic solvent, selected from methanol, ethanol, isopropanol, etc., with methanol being preferred; the acidic catalyst used is selected from formic acid, acetic acid, hydrochloric acid, etc., and the temperature of condensation reaction is 50-80℃.

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