Synthetic method of naphthalimide derivative

Abstract

The invention relates to the technical field of naphthalimide preparation, in particular to a synthetic method of naphthalimide derivatives. Comprising the following steps: dissolving a mixture of 1, 8-naphthalic anhydride and 4-aminoacetophenone in acetic acid containing sodium acetate, carrying out a reflux reaction, after the reaction is completed, cooling the mixture to room temperature, filtering to obtain a crude product, and washing to obtain a pure compound 1; the method comprises the following steps: dissolving a mixture of a compound 1, a hydrazine derivative and an acid catalyst in an organic solvent, carrying out a reflux reaction, after the reaction is completed, cooling the mixture to room temperature, filtering to obtain a crude product, and washing to obtain the naphthalimide compound. The synthesis reaction is safe and reliable, the process is simple and easy to control, and the reaction yield is high; the combination of the synthesized drug and DNA has spontaneity, and the drug has good anti-proliferative activity.

Description

Technical Field

[0001] This invention relates to the field of naphthalimide preparation technology, specifically to a method for synthesizing naphthalimide derivatives. Background Technology

[0002] Malignant tumors, as a major disease posing a serious threat to human life and health, have always been a research hotspot in the fields of medicine and pharmacy. Chemotherapy, as one of the main methods of tumor treatment, relies on chemical drugs with specific anti-tumor activity. Among them, anticancer drugs that target DNA have become an important direction in anti-tumor drug development because they can directly interfere with the replication and expression of genetic material in tumor cells.

[0003] Naphthalene imide compounds, with their unique planar conjugated structure, can specifically intercalate with DNA molecules. By intercalating into the DNA double helix and disrupting the normal structure and function of DNA, they inhibit the proliferation of tumor cells, thus exhibiting significant antitumor activity and attracting much attention in the field of anticancer drug research. Among these compounds, mitonafide and amonafide are two representative naphthalene imide drugs, which have shown good antitumor effects in multiple clinical trials and exhibited strong inhibitory activity against various tumor cell lines (Liang GB, et al. European Journal of Medicinal Chemistry, 2021, 210: 112951.).

[0004] However, the aforementioned two naphthylimide drugs have revealed serious limitations in clinical application, greatly hindering their further development and application. Both mitonaphthylamine and aminonaphthylamine have demonstrated severe central nervous system toxicity in clinical trials. This neurotoxicity leads to a series of adverse reactions in patients, such as headache, dizziness, and cognitive impairment, severely impacting patients' quality of life and treatment adherence. Simultaneously, both drugs cause significant venous inflammation during administration, not only causing additional pain for patients but also potentially making the dosing regimen impossible due to venous irritation. Due to these side effects, the clinical trials of aminonaphthylamine are currently stalled in Phase III, failing to advance to market application; while the Phase II clinical trial of mitonaphthylamine has failed, preventing its continued clinical application. These results indicate that the toxic side effects of existing naphthalimide drugs have become a key bottleneck restricting their clinical translation, and there is an urgent need to improve their safety and tolerability through structural modification and optimization (Parish CA, et al. Journal of Natural Products, 2009, 72(1): 59. and Banerjee S, et al. Chemical Society Reviews, 2013, 42(4): 1601.).

[0005] Structural modification based on the basic skeleton of naphthalimide compounds is an important research strategy for improving their pharmacological activity and reducing toxic side effects. By introducing different functional side chains or aromatic systems, the binding ability of compounds to DNA can be adjusted, their physicochemical properties and pharmacokinetic behavior can be improved, thereby achieving the goal of reducing toxicity and improving efficacy. The hydrazone group (-C=N-NH-) is an important functional active group with diverse biological activities and has broad application value in the field of medicinal chemistry. For example, the compound A-007, containing a hydrazone structure, exhibits high cytotoxicity against human breast cancer cell lines (MCF-7 and ZR-75-1), showing good potential for anti-tumor applications (Nasr T, et al. European Journal of Medicinal Chemistry, 2018, 151: 723-739.). In addition, the hydrazone group has strong nucleophilicity, making it easy to coordinate with metal ions, which may enhance the bioactivity of drugs by regulating the distribution and function of metal ions in the body. At the same time, the -NH- part in the hydrazone group can interact with suitable anions and biomolecules (such as proteins and nucleic acids) in the body, which helps to improve the specific binding ability of drugs to targets and reduce the toxic side effects caused by non-specific effects (Chang HQ, et al. Transition Metal Chemistry, 2015, 40(5):485-491. and Qin JC, et al. Journal of Photochemistry and Photobiology A:Chemistry, 2016, 324: 152-158.). Summary of the Invention

[0006] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for synthesizing naphthalimide derivatives. The synthesis reaction is safe and reliable, the process is simple and easy to control, and the reaction yield is high. The synthesized drug binds to DNA spontaneously and has good anti-proliferative activity.

[0007] This invention is achieved using the following technical solution: The method for synthesizing the aforementioned naphthalimide derivatives includes the following steps: (1) Dissolve a mixture of 1,8-naphthalenedicarboxylic anhydride and 4-aminoacetophenone in acetic acid containing sodium acetate and reflux the reaction. After the reaction is complete, cool the mixture to room temperature and filter to obtain the crude product. Wash the crude product to obtain pure compound 1. (2) The mixture of compound 1, hydrazine derivative and acid catalyst was dissolved in an organic solvent and refluxed. After the reaction was completed, the mixture was cooled to room temperature and filtered to obtain crude product. The crude product was washed to obtain naphthalimide compounds.

[0008] In step (1), the molar ratio of 1,8-naphthalenedicarboxylic anhydride and 4-aminoacetophenone is (1-1.5):(1-1.5), preferably 1:1; the molar ratio of sodium acetate to 1,8-naphthalenedicarboxylic anhydride is 1:(2-2.5), preferably 1:2.

[0009] In step (1), acetic acid is used as a solvent, and the reflux reaction time is 24-36 h, preferably 24 h; the solvent used for washing is ethanol.

[0010] The hydrazine derivative in step (2) is one of the acylhydrazine compounds.

[0011] The hydrazine derivative in step (2) is one of p-methylbenzenesulfonyl hydrazine, nicotinic hydrazine, or methoxyacetyl hydrazine.

[0012] The molar ratio of compound 1 and hydrazine derivative in step (2) is 1:(5.5-6), preferably 1:5.5.

[0013] The acid catalyst in step (2) is one of glacial acetic acid, sulfuric acid or hydrogen chloride, and the amount of acid catalyst is 0.5% to 500% of 1 mole of the compound. The organic solvent is one of 1,2-dichloroethane or anhydrous ethanol.

[0014] The reflux reaction time in step (2) is 6-8 hours, preferably 6 hours, and the solvent used for washing is ethyl acetate.

[0015] A mixture of 1,8-naphthalenedicarboxylic anhydride and 4-aminoacetophenone was dissolved in acetic acid containing sodium acetate and refluxed for 24 hours. After the reaction was complete (TLC), the mixture was cooled to room temperature and filtered to give a crude product, which was then washed with ethanol to give pure compound 1 as a yellow solid.

[0016] .

[0017] A mixture of compound 1, 4-methylbenzenesulfonylhydrazine, and glacial acetic acid as a catalyst was dissolved in 1,2-dichloroethane and refluxed for 6 hours. After the reaction was complete (TLC), the mixture was cooled to room temperature and filtered to give a crude product, which was then washed with ethyl acetate (3 × 15 mL) to give pure compound 2 as a white solid, N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylene)-4-methylbenzenesulfonylhydrazine.

[0018] Compound 1, nicotinic acid hydrazide, and glacial acetic acid were placed in anhydrous ethanol, and compound 3 was prepared following the same procedure as compound 2. Pure compound 3 was obtained as a white solid, N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylidene)nicotinic acid hydrazide.

[0019] Compound 4 was prepared from compound 1, 2-methoxyacetylhydrazine, acetic acid, and anhydrous ethanol as solvent, following the steps described for compound 2. The pure compound 4 was obtained as a yellow solid, N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylidene)-2-methoxyacetylhydrazine.

[0020] .

[0021] Compared with the prior art, the beneficial effects of the present invention are: (1) The synthesis reaction of the present invention is safe and reliable, the process is simple and easy to control, and the reaction yield is high.

[0022] (2) The drug synthesized in this invention binds to DNA spontaneously and has good anti-proliferative activity. Attached Figure Description

[0023] Figure 1 This is the proton NMR spectrum of compound 1 in Example 1 of the present invention; Figure 2 The carbon spectrum of compound 1 in Example 1 of this invention; Figure 3 The hydrogen spectrum of compound 2 in Example 1 of this invention; Figure 4 The carbon spectrum of compound 2 in Example 1 of this invention; Figure 5 This is a high-resolution mass spectrum of compound 2 in Example 1 of the present invention; Figure 6 The hydrogen spectrum of compound 3 in Example 2 of this invention; Figure 7 This is the high-resolution mass spectrum of compound 3 in Example 2 of the present invention; Figure 8 The hydrogen spectrum of compound 4 in Example 2 of this invention; Figure 9 This is the high-resolution mass spectrum of compound 4 in Example 2 of the present invention; Figure 10 The UV-Vis spectrum is shown for the titration of 0 to 400 μL of ctDNA (200 μg / mL) with 200 μM compound 2 (A) at 37 °C. Figure 11The UV-Vis spectrum is shown for the titration of 0 to 400 μL of ctDNA (200 μg / mL) with 200 μM compound 3 (B) at 37 °C. Figure 12 The UV-Vis spectra of 200 μM compound 4 (C) titrated with 0 to 400 μL of ctDNA (200 μg / mL) at 37 °C. Figure 13 The linear fitting curve of A0 / (A0-A) versus 1 / [DNA] for compound 2(A); Figure 14 The linear fitting curve of A0 / (A0-A) versus 1 / [DNA] for compound 3(B); Figure 15 The linear fitting curve of A0 / (A0-A) versus 1 / [DNA] for compound 4 (C); Figure 16 The cell viability (%) of A549 cells after treatment with mitonaphthylamine, aminonaphthylphenate, compound 2 (N-1), compound 3 (N-2), and compound 4 (N-3) for 24 h. Detailed Implementation

[0024] To make the objectives and technical solutions of this invention clearer, the invention will be further described in detail below.

[0025] Example 1 (1) A mixture of 1,8-naphthalenedicarboxylic anhydride (1.0 mmol, 0.198 g) and 4-aminoacetophenone (1.0 mmol, 0.136 g) was dissolved in acetic acid (15 mL) containing sodium acetate (2.0 mmol, 0.165 g) and refluxed for 24 hours. After the reaction was complete (TLC), the mixture was cooled to room temperature and filtered to give a crude product, which was then washed with ethanol to give pure compound 1 (0.170 g) as a yellow solid, yield 54.0%, mp > 300 °C.

[0026] (2) A mixture of compound 1 (1.0 mmol, 0.315 g), 4-methylbenzenesulfonyl hydrazine (5.5 mmol, 1.023 g), and glacial acetic acid (5.0 mmol, 0.286 mL) as a catalyst was dissolved in 1,2-dichloroethane (7 mL) and refluxed for 6 hours. After the reaction was complete (TLC), the mixture was cooled to room temperature and filtered to obtain a crude product, which was then washed with ethyl acetate (3 × 15 mL) to obtain pure compound 2 (0.357 g) as a white solid with a yield of 74.0% and a melting point of 260-261 °C. Compound 2 is N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylidene)-4-methylbenzenesulfonyl hydrazine.

[0027] Compound 1 was tested, and the proton NMR spectrum was obtained: 1 H NMR (CDCl3, 500 MHz): δ 8.70(2H, d, -Ar),8.34(2H, d, -Ar), 8.19(2H, d, -Ar), 7.85(2H, t, -Ar), 7.49(2H, d, -Ar), 2.71(3H, s, -COC H 3). Carbon spectrum: 13 C NMR (CDCl3, 125 MHz): δ 196.2, 163.1, 138.8, 136.1, 133.6, 130.8, 128.4, 128.2, 127.5, 126.1, 121.5, 25.8. Figure 1 The hydrogen spectrum of compound 1, Figure 2 This is the carbon spectrum of compound 1. Compound 2 was tested: 1H NMR spectrum: 1 H NMR (DMSO- d 6, 500 MHz): δ 10.58(1H, s, =NN H -), 8.51(4H, ddd, -Ar), 7.91(2H, dd, -Ar), 7.87–7.82(2H, m, -Ar), 7.80–7.75(2H, m, -Ar), 7.45–7.39(4H, m, -Ar), 2.38(3H, s, -N=CC H 3), 2.25(3H, s, -Ar-CC H 3). Carbon spectrum: 13 C NMR (DMSO- d (6, 125 MHz): δ 164.1, 153.1, 143.9, 137.7, 137.5, 136.6, 135.0, 131.9, 131.2, 130.0, 128.3, 128.1, 127.7, 126.9, 123.0, 21.5, 14.9. High-resolution mass spectrometry: HRMS calculated for C 20 H 13 NO3[M+H] + : 484.1331, found 484.1335. Figure 3 The hydrogen spectrum of compound 2, Figure 4 The carbon spectrum of compound 2. Figure 5 This is the high-resolution mass spectrum of compound 2.

[0028] Example 2 Step (1) is the same as in Example 1.

[0029] (2) Compound 1 (1.0 mmol, 0.315 g), nicotinamide (5.5 mmol, 0.686 g), and glacial acetic acid (5.0 mmol, 0.286 mL) were placed in anhydrous ethanol (8 mL), and compound 3 was prepared according to the preparation steps of compound 2. Pure compound 3 (0.165 g) was obtained as a white solid with a yield of 38.0% and a melting point >300 °C. Compound 3 is N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylidene)nicotinamide.

[0030] Tests were performed on compound 3: 1H NMR spectrum: 1 H NMR (DMSO- d 6, 500MHz): δ 11.05(1H, s, =NN H -), 9.07(1H, s, O=CCC H =N-), 8.77(1H, s, -NC H =CH-), 8.53(4H, d, -Ar),8.26(1H, s, O=CC=C H -), 8.05–7.89(4H, m, -Ar), 7.57(1H, s, =N-CH=C H -), 7.48(2H, d, -Ar), 2.47(3H, s, -Ar-CC H 3). High-resolution mass spectrometry: HRMS calculated for C 26 H 18 N4O3[M+H] + : 435.1457, found 435.1447. Figure 6 The hydrogen spectrum of compound 3. Figure 7 This is the high-resolution mass spectrum of compound 3.

[0031] Example 3 Step (1) is the same as in Example 1.

[0032] (2) Compound 4 was prepared from compound 1 (1.0 mmol, 0.315 g), 2-methoxyacetylhydrazine (5.5 mmol, 0.572 g), acetic acid (5.0 mmol, 0.286 mL), and anhydrous ethanol (8 mL) as solvent, following the steps described for compound 2. Pure compound 4 (0.136 g) was obtained as a yellow solid with a yield of 34.0% and a melting point of 258-259 °C. Compound 4 is N-(1-(4-(1,3-dioxo-1H-benzo[de]isoquinoline-2(3H)-yl)phenyl)ethylidene)-2-methoxyacetylhydrazine.

[0033] Tests were performed on compound 4: 1H NMR spectrum: 1 H NMR (DMSO- d 6, 500 MHz) δ 10.61(1H, s, =NN H -),8.41(4H, d, -Ar), 7.82–7.78(4H, m, -Ar), 7.33(2H, d, -Ar), 4.39(2H, s, O=CC H 2-O-), 2.20(3H, s, -OC H 3), 1.12(3H, s, -Ar-CC H 3). High-resolution mass spectrometry: HRMS calculated for C 23 H 19 N3O4[M+H] + : 402.1454, found 402.1449. Figure 8 The proton spectrum of compound 4. Figure 9 This is the high-resolution mass spectrum of compound 4.

[0034] Figure 10-12 The UV-Vis spectra of 200 μM compounds 2 (A), 3 (B), and 4 (C) titrated with 0 to 400 μL of ctDNA (200 μg / mL) at 37 °C are shown. The intercalation mechanisms of compounds 2, 3, and 4 with ctDNA were tested using UV-Vis spectroscopy. When drug molecules insert into double-stranded DNA, the π* orbitals of the drug molecules couple with the π orbitals of the DNA base pairs, reducing the π-π* transition energy and shifting the UV-Vis absorption wavelength to a longer wavelength (redshift). Simultaneously, the coupling of partially filled π orbitals reduces the transition probability, resulting in a heterochromatic effect. Figure 10-12 As shown, the three drugs were added sequentially. ctThe absorbance of the DNA-drug mixture changes. With increasing ctDNA addition, the maximum UV absorption peak of the drug decreases, and the maximum absorption wavelength also red-shifts, exhibiting a color reduction effect. According to the formula: The calculated decolorization rates (H) of compounds 2, 3, and 4 were 25.13%, 15.10%, and 22.50%, respectively. (Compound 2-) ct DNA, compound 3- ct DNA and compounds 4- ct The binding constant of DNA complexes (K) b Based on the formula: Where A0 and A are respectively the result of not adding ct Compound 2, compound 3, or compound 4 and added to DNA ct The uptake value of the complex formed after DNA, ε G and ε H-G These are compound 2, compound 3 or compound 4, and added ct The molar extinction coefficient of the complex formed after DNA, [DNA] is ct DNA concentration. The A0 / (A0-A) value is linearly related to 1 / [DNA]. Figure 13-15 The ratio of intercept to slope is K. b Compound 2- ct K of DNA complex b 2.42×10 4 M -1 Compound 3- ct K in DNA complex b It is 1.67 × 10 4 M -1 Compound 4- ct K in DNA complex b 1.18×10 4 M -1 In addition, compound 2- ct DNA, compound 3- ct DNA and compounds 4- ctDNA The stability of the complex was also calculated using the following formula: Calculate the change in Gibbs free energy (ΔG). The calculated Gibbs free energy is less than zero (ΔG<0), indicating that the intercalation pattern between the molecule and ctDNA is spontaneous.

[0035] like Figure 16As shown, the inhibitory activity of compounds 2 (N-1), 3 (N-2), and 4 (N-3) against human non-small cell lung cancer cells A549 was evaluated using the MTT assay, with mitonaphthylamine and aminaflipide as positive controls. For the A549 cell line, the concentration of compound 2 (N-1), compound 3 (N-2), or compound 4 (N-3) was 50 μg·mL⁻¹. -1 At these concentrations, cell viability was 35.12%, 39.00%, and 41.07%, respectively. When the concentration of N-1, N-2, or compound 4 (N-3) reached 100 μg / mL... -1 At the specified times, the cell viability was 24.12%, 26.71%, and 32.55%, respectively, lower than that of mitonaphthylamine (44.35%) and aminofivir (49.00%). Furthermore, compounds 2 (N-1), 3 (N-2), and 4 (N-3) showed low IC50 values ​​for A549 cell lines. 50 The values ​​were 28.73 µg·mL. -1 27.91µg·mL -1 and 28.42 µg·mL -1 This indicates that compounds 2 (N-1), 3 (N-2), and 4 (N-3) all exhibited superior antiproliferative activity compared to mitonaphthylamine (55.36 µg·mL⁻¹). -1 ) and aminapeptide (63.61 µg·mL -1 Therefore, compounds 2 (N-1), 3 (N-2), and 4 (N-3) achieved the expected inhibitory effects on cancer cells, which may be due to the introduction of hydrazone groups enhancing intercalation ability and oxidative damage to DNA. Mitonaphthylamine, aminonaphthylphenate, compounds 2 (N-1), 3 (N-2), and 4 (N-3) showed IC50 values ​​against the A549 cell line. 50 The values ​​are shown in Table 1.

[0036] Table 1 shows the IC50 values ​​for the A549 cell line. 50 value

Claims

1. A method for synthesizing a naphthalimide derivative, characterized by, The method comprises the following steps: (1) dissolving a mixture of 1,8-naphthalic anhydride and 4-aminoacetophenone in acetic acid containing sodium acetate, refluxing, cooling the mixture to room temperature after the reaction is completed, and filtering to obtain a crude product, and washing to obtain a pure compound 1; (2) dissolving a mixture of the compound 1, a hydrazine derivative, and an acid catalyst in an organic solvent, refluxing, cooling the mixture to room temperature after the reaction is completed, and filtering to obtain a crude product, and washing to obtain a naphthalimide compound.

2. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, In the step (1), the molar ratio of 1,8-naphthalic anhydride to 4-aminoacetophenone is (1-1.5):(1-1.5); and the molar ratio of sodium acetate to 1,8-naphthalic anhydride is 1:(2-2.5).

3. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, In the step (1), the refluxing time is 24-36 h; the reaction solvent is acetic acid; and the washing solvent is ethanol.

4. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, The hydrazine derivative in the step (2) is one of acylhydrazine compounds.

5. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, The hydrazine derivative in the step (2) is one of p-methylbenzenesulfonylhydrazide, nicotinoylhydrazine, or methoxyacetylhydrazine.

6. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, In the step (2), the molar ratio of the compound 1 to the hydrazine derivative is 1:(5.5-6).

7. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, The acid catalyst in the step (2) is one of glacial acetic acid, sulfuric acid, or hydrogen chloride; the amount of the acid catalyst is 0.5%-500% of the molar amount of the compound 1; and the organic solvent is one of 1,2-dichloroethane or anhydrous ethanol.

8. The method of synthesis of naphthalimide derivatives according to claim 1, wherein, In the step (2), the refluxing time is 6-8 h; and the washing solvent is ethyl acetate.

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