A spirocyclic barbituric acid derivative and its preparation method and application

Abstract

The present invention relates to a spirocyclic barbituric acid derivative and a preparation method and application thereof. The present invention uses a tryptamine isonitrile derivative and an unsaturated barbituric acid derivative as reaction raw materials, and can achieve accurate synthesis of the spirocyclic barbituric acid derivative through a simple heating reaction. The preparation process of the spirocyclic barbituric acid derivative of the present invention is simple, the atom economy is high, and the post-processing is convenient. The obtained spirocyclic barbituric acid derivative has potential drug activity, shows significant inhibitory effects on a variety of cancer cells, and has broad application prospects in the preparation of anti-tumor drugs.

Description

Technical Field

[0001] The invention relates to the technical field of organic synthesis, in particular to a spirocyclic barbituric acid derivative and a preparation method and application thereof. Background Art

[0002] The activity of barbiturates is closely related to their molecular structure, especially the nature and position of the substituents, which affect the pharmacological action, potency, and duration of action of the drugs. The mechanism of action of this class of drugs usually involves the regulation of γ-aminobutyric acid (GABA) receptors in the central nervous system, thereby enhancing the opening of GABA-mediated chloride channels, leading to neuronal hyperpolarization and reducing neuronal excitability. Barbituric acid derivatives are attractive barbiturates because they are commonly found in pharmacologically active compounds. In addition, spirobarbiturates have continued to attract attention in recent years due to their various pharmacological properties, such as their applications in urease inhibitors, HIV-1 inhibitors, anticonvulsants, TACE inhibitors, MMP-13 inhibitors, and anticancer activities (Adv. Synth. Catal. 2016, 358, 2619 –2630; Org. Biomol. Chem., 2019,17, 9200–9208; Chem. Biodiversity. 2022, 19, e202100809; Adv. Synth. Catal.2024, 366, 1–7). Therefore, strategies for constructing spirobarbituric acid compounds are frequently reported, and the structures of common spirobarbituric acid derivatives are shown below:

[0003] .

[0004] Although there have been several reports on the synthesis of the above compounds, these methods require multiple additives, use expensive metal catalysts, and cannot control the product configuration. They are very inefficient in terms of step economy and atom economy. In the preparation of spirobarbituric acid derivatives, synthesis efficiency is the most important part. In addition, simple operation, safety, economy, and environmental protection also need to be taken into account to achieve the requirements of the "green organic synthesis concept." Therefore, under mild conditions, starting from simple and readily available substrates, it is extremely attractive to efficiently construct a precise synthesis method for spirobarbituric acid derivatives. At the same time, it is also of great significance to synthesize biologically active spirobarbituric acid derivatives by simple thermal reaction synthesis methods without using expensive metal catalysts in the synthesis process. Summary of the invention

[0005] Therefore, the technical problem to be solved by the present invention is to overcome the lack of a simple method for preparing spirocyclic barbituric acid derivatives in the prior art.

[0006] In order to solve the above technical problems, the present invention provides a spirocyclic barbituric acid derivative and a preparation method and application thereof. The present invention uses a tryptamine isonitrile derivative and an unsaturated barbituric acid derivative as reaction raw materials to achieve the precise synthesis of the spirocyclic barbituric acid derivative. The preparation process of the spirocyclic barbituric acid derivative of the present invention is simple, the atom economy is high, and the post-processing is convenient. The obtained spirocyclic barbituric acid derivative has potential pharmaceutical activity.

[0007] The first object of the present invention is to provide a spirocyclic barbituric acid derivative, the general structural formula of the spirocyclic barbituric acid derivative is shown in Formula III:

[0008] ;

[0009] Among them, R 1 is selected from hydrogen, alkyl or halogen;

[0010] R 2 is selected from hydrogen or alkyl;

[0011] R 3 is selected from hydrogen, halogen, alkyl or nitro.

[0012] Furthermore, the halogen includes chlorine or bromine.

[0013] Furthermore, the alkyl group includes a C1-C3 straight chain alkyl group or a C3-C6 cycloalkyl group.

[0014] Furthermore, the structure of the spirocyclic barbituric acid derivative is shown in Formula III1-III9:

[0015] .

[0016] The second object of the present invention is to provide a method for preparing the above-mentioned spirocyclic barbituric acid derivatives, wherein a tryptamine isocyanide derivative and an unsaturated barbituric acid derivative are subjected to a heating reaction in the presence of an organic solvent to prepare the spirocyclic barbituric acid derivative; wherein the general structural formula of the tryptamine isocyanide derivative is as shown in Formula I:

[0017] ;

[0018] Among them, R 1 is selected from hydrogen, alkyl or halogen;

[0019] The general structural formula of the unsaturated barbituric acid derivative is shown in Formula II:

[0020] ;

[0021] Among them, R 2 is selected from hydrogen or alkyl;

[0022] R 3 is selected from hydrogen, halogen, alkyl or nitro.

[0023] Furthermore, the organic solvent is selected from one or more of acetonitrile, methanol, ethanol, toluene, acetone, triethylamine, tetrahydrofuran, 1,2-dichloroethane, 1,4-dioxane, dimethyl sulfoxide and N,N-dimethylformamide.

[0024] Furthermore, the temperature of the heating reaction is 30-90° C., and the time of the heating reaction is 6-14 hours.

[0025] Furthermore, the structures of the tryptamine isonitrile derivatives are shown in formulas (1) to (4):

[0026] .

[0027] Furthermore, the structure of the unsaturated barbituric acid derivative is shown in formula (5)-(10):

[0028] .

[0029] Further, taking the reaction in which the raw materials are tryptamine isonitrile, unsaturated barbituric acid, and the solvent is dimethyl sulfoxide as an example, the reaction principle of the spirocyclic barbituric acid derivative represented by formula III of the present invention during the preparation process is as follows:

[0030] .

[0031] Tryptamine isocyanide Ⅰ undergoes Michael addition reaction with unsaturated barbituric acid compound Ⅱ to generate intermediate A, and then tryptamine isocyanide indole C3 position cyclization generates intermediate B, then intermediate B double bond shifts to obtain intermediate C, and finally undergoes cyclization reaction with tryptamine isocyanide C2 position to obtain the final product III.

[0032] The third object of the present invention is to provide a use of the above-mentioned spirocyclic barbituric acid derivative in the preparation of anti-tumor active drugs, and the types of tumors include breast cancer, human oral epidermoid carcinoma and cervical cancer.

[0033] Beneficial effects of the present invention:

[0034] The present invention uses tryptamine isonitrile derivatives and unsaturated barbituric acid derivatives as reaction raw materials, and can achieve accurate synthesis of spirocyclic barbituric acid derivatives through a simple heating reaction. The preparation process of the spirocyclic barbituric acid derivatives of the present invention is simple, the atom economy is high, and the post-processing is convenient. The obtained spirocyclic barbituric acid derivatives have potential drug activity, show significant inhibitory effects on a variety of cancer cells, and have broad application prospects in the preparation of anti-tumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is the synthetic route of the spirocyclic barbituric acid derivatives of the present invention. DETAILED DESCRIPTION

[0036] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0037] Example 1: Synthesis of the compound shown in (III1)

[0038] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of unsaturated barbituric acid (compound corresponding to number (5), 0.0272 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0337 g of reaction product.

[0039] The above reaction products were characterized and the results were as follows:

[0040] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.49-7.32 (m, 6H), 7.20 (d, J =7.6Hz, 1H), 7.00(t, J =7.6Hz, 1H), 6.63(t, J =7.6Hz, 1H), 6.54 (s, 1H), 6.44 (d, J =8.0 Hz, 1H), 4.17 (s, 1H), 4.10-4.03 (m, 1H), 3.83-3.76 (m, 1H), 3.13 (s, 3H), 3.06 (s, 3H), 2.20-2.11 (m, 1H), 1.89-1.82 (m, 1H) ppm.

[0041] According to the characterization data, the obtained reaction product is pure (purity>95%); the product yield is calculated and the result is 76%.

[0042] Example 2: Synthesis of the compound shown in (III2)

[0043] 0.1 mmol of 5-chlorotryptamine isonitrile (compound corresponding to number (2), 0.0204 g) and 0.1 mmol of unsaturated barbituric acid (compound corresponding to number (5), 0.0272 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0298 g of reaction product.

[0044] The above reaction products were characterized and the results were as follows:

[0045] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.55-7.24 (m, 6H), 7.18-7.12 (m, 1H), 7.03 (dd, J =8.4, 2.0Hz, 1H), 6.75 (s, 1H), 6.43 (d, J =8.4Hz, 1H), 4.22 (s, 1H), 4.11-4.02 (m, 1H), 3.86-3.74 (m, 1H), 3.12 (s, 3H), 3.06 (s, 3H), 2.22-2.09 (m, 1H), 1.94-1.85 (m, 1H) ppm.

[0046] According to the characterization data, the obtained reaction product is pure (purity> 95%); the product yield is calculated and the result is 62%.

[0047] Example 3: Synthesis of the compound shown in (III3)

[0048] 0.1 mmol of 5-bromotryptamine isonitrile (compound corresponding to number (3), 0.0248 g) and 0.1 mmol of unsaturated barbituric acid (compound corresponding to number (5), 0.0272 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0366 g of reaction product.

[0049] The above reaction products were characterized and the results were as follows:

[0050] 1H NMR (400 MHz, DMSO- d 6 ) d 7.63-7.08 (m, 8H), 6.79 (s, 1H), 6.40 (d, J =8.4Hz, 1H), 4.22 (s, 1H), 4.14-3.98 (m, 1H), 3.88-3.71 (m, 1H), 3.12 (s, 3H), 3.06 (s, 3H), 2.23-2.09 (m, 1H), 1.97-1.81 (m, 1H) ppm.

[0051] According to the characterization data, the obtained reaction product is pure (purity>95%); the product yield is calculated and the result is 70%.

[0052] Example 4: Synthesis of the compound shown in (III4)

[0053] 0.1 mmol of 5-methyltryptamine isocyanate (compound corresponding to number (4), 0.0184 g) and 0.1 mmol of unsaturated barbituric acid (compound corresponding to number (5), 0.0272 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0212 g of reaction product.

[0054] The above reaction products were characterized and the results were as follows:

[0055] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.61-7.34 (m, 7H), 7.02 (s, 1H), 6.81 (d, J =8.0Hz, 1H), 6.34(d, J =8.0Hz, 1H), 4.15 (s, 1H), 4.11-4.05 (m, 1H), 3.92-3.88 (m, 1H), 3.10 (s, 3H), 3.04 (s, 3H), 2.24 (s, 3H), 2.17-2.10 (m, 1H), 1.89-1.82 (m, 1H) ppm.

[0056] According to the characterization data, the obtained reaction product is pure (purity> 95%); the product yield is calculated and the result is 46%.

[0057] Example 5: Synthesis of the compound shown in (III5)

[0058] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of 4-nitrobenzoylbarbituric acid (compound corresponding to number (6), 0.0317 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1) to obtain 0.0275 g of the reaction product.

[0059] The above reaction products were characterized and the results were as follows:

[0060] 1 H NMR (400 MHz, DMSO- d 6 ) d 8.25-7.51 (m, 6H), 7.20 (d, J =7.6Hz, 1H), 7.01(t, J =7.6Hz, 1H), 6.64(t, J =7.2Hz, 1H), 6.45(d, J =8.0Hz, 1H), 4.19 (s, 1H), 4.14-3.94 (m, 2H), 3.35-3.10 (m, 6H), 2.23-2.10 (m, 1H), 1.92-1.80 (m, 1H) ppm.

[0061] According to the characterization data, the obtained reaction product is pure (purity>95%); the product yield is calculated and the result is 56%.

[0062] Example 6: Synthesis of the compound shown in (III6)

[0063] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of 4-chlorobenzoyl unsaturated barbituric acid (compound corresponding to number (7), 0.0306 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0249 g of reaction product.

[0064] The above reaction products were characterized and the results were as follows:

[0065] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.58-7.29 (m, 6H), 7.19 (d, J =7.6Hz, 1H), 7.00(t, J =7.6Hz, 1H), 6.63(t, J =7.2Hz, 1H), 6.43(d, J =8.0Hz, 1H), 4.17 (s, 1H), 4.11-4.02 (m, 1H), 3.80-3.73 (m, 1H), 3.17 (s, 3H), 3.05 (s, 3H), 2.20-2.09 (m, 1H), 1.91-1.82 (m, 1H) ppm.

[0066] According to the characterization data, the obtained reaction product is pure (purity> 95%); the product yield is calculated and the result is 52%.

[0067] Example 7: Synthesis of the compound shown in (III7)

[0068] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of 4-bromobenzoyl unsaturated barbituric acid (compound corresponding to number (8), 0.0350 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0324 g of reaction product.

[0069] The above reaction products were characterized and the results were as follows:

[0070] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.69-7.26 (m, 6H), 7.19 (d, J =7.2Hz, 1H), 6.99(t, J =8.0Hz, 1H), 6.63(t, J =7.4Hz, 1H), 6.44(d, J=7.6Hz, 1H), 4.17 (s, 1H), 4.12-4.00 (m, 1H), 3.82-3.73 (m, 1H), 3.18 (s, 3H), 3.05 (s, 3H), 2.20-2.09 (m, 1H), 1.91-1.81 (m, 1H) ppm.

[0071] According to the characterization data, the obtained reaction product is pure (purity> 95%); the product yield is calculated and the result is 62%.

[0072] Example 8: Synthesis of the compound shown in (III8)

[0073] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of 4-methylbenzoyl unsaturated barbituric acid (compound corresponding to number (9), 0.0268 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0231 g of reaction product.

[0074] The above reaction products were characterized and the results were as follows:

[0075] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.37-7.15 (m, 6H), 7.00 (t, J =7.6Hz, 1H), 6.63(t, J =7.6Hz, 1H), 6.55-6.48 (t, J =7.6Hz, 1H), 6.43(d, J =8.0Hz, 1H), 4.15 (s, 1H), 4.11-4.00 (m, 1H), 3.83-3.73 (m, 1H), 3.15 (s, 3H), 3.05 (s, 3H), 2.33 (s, 3H), 2.19-2.08 (m, 1H), 1.89-1.81 (m, 1H) ppm.

[0076] According to the characterization data, the obtained reaction product is a pure product (purity> 95%); the product yield is calculated and the result is 50%.

[0077] Example 9: Synthesis of the compound shown in (III9)

[0078] 0.1 mmol of tryptamine isonitrile (compound corresponding to number (1), 0.0170 g) and 0.1 mmol of 2-methylbenzoyl unsaturated barbituric acid (compound corresponding to number (10), 0.0268 g) were weighed into an 8 mL reaction bottle, 1 mL of dimethyl sulfoxide was added as solvent, and the mixture was stirred at 50°C for 12 hours. After the reaction, the reaction solution was subjected to vacuum evaporation and column chromatography separation (column chromatography separation conditions: stationary phase was 200-300 mesh silica gel powder, mobile phase was dichloromethane (A) and anhydrous methanol (B), and the mobile phase change program A:B was 50:1), to obtain 0.0322 g of reaction product.

[0079] The above reaction products were characterized and the results were as follows:

[0080] 1 H NMR (400 MHz, DMSO- d 6 ) d 7.42-6.95 (m, 7H), 6.62-6.54 (m, 2H), 6.41 (d, J =8.0Hz, 1H), 4.18 (s, 1H), 3.95 (s, 1H), 3.75 (s, 1H), 3.23 (s, 3H), 3.03 (s, 3H), 2.21 (s, 1H), 2.13 (s, 3H), 1.88-1.75 (m, 1H) ppm.

[0081] According to the characterization data, the obtained reaction product is pure (purity>95%); the product yield is calculated and the result is 70%.

[0082] The present invention selected the 9 compounds prepared in the above examples and used the MTT method to perform toxicity tests on three human cancer cells (where pTACE is tumor necrosis factor (TNFa) converting enzyme, a-glucosidase is α-glucosidase, and HT-29 is a human colorectal adenocarcinoma cell), and used the drug oxaliplatin as a toxicity test control. The results are shown in Table 1. The results show that the compounds represented by formula III2, III3, III5, III6, and III7 exhibit low micromolar cytotoxicity; the compound represented by formula III1 is not active (IC 50 >100μM). It is noteworthy that spirobarbituric acid compounds III5 and III6 showed stronger inhibitory effects on one or more human cancer cells (IC 50 lower than oxaliplatin).

[0083] Table 1 Cytotoxicity (IC) of different compounds 50 , μM) test results

[0084]

[0085] The above results indicate that the spirocyclobarbituric acid derivatives synthesized in the present invention have anti-tumor activity and can be prepared into anti-tumor drugs.

[0086] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. A spirocyclic barbituric acid derivative, characterized in that: The general structural formula of the spirocyclic barbituric acid derivative is shown in Formula III: Among them, R 1 is selected from hydrogen, alkyl or halogen; R 2 is selected from hydrogen or alkyl; R 3 is selected from hydrogen, halogen, alkyl or nitro; The alkyl group includes a C1-C3 straight chain alkyl group or a C3-C6 cycloalkyl group.

2. The spirocyclic barbituric acid derivative according to claim 1, characterized in that The halogen is chlorine or bromine.

3. The spirocyclic barbituric acid derivative according to claim 1, characterized in that The structures of the spirocyclic barbituric acid derivatives are shown in Formula III1-III9:

4. A method for preparing a spirocyclic barbituric acid derivative according to any one of claims 1 to 3, characterized in that: The spirocyclic barbituric acid derivative is prepared by heating a tryptamine isocyanide derivative and an unsaturated barbituric acid derivative in the presence of an organic solvent; wherein the structural formula of the tryptamine isocyanide derivative is as shown in Formula I: Among them, R 1 is selected from hydrogen, alkyl or halogen; The general structural formula of the unsaturated barbituric acid derivative is shown in Formula II: Among them, R 2 is selected from hydrogen or alkyl; R 3 is selected from hydrogen, halogen, alkyl or nitro.

5. The preparation method according to claim 4, characterized in that: The organic solvent is selected from one or more of acetonitrile, methanol, ethanol, toluene, acetone, triethylamine, tetrahydrofuran, 1,2-dichloroethane, 1,4-dioxane, dimethyl sulfoxide and N,N-dimethylformamide.

6. The preparation method according to claim 4, characterized in that: The temperature of the heating reaction is 30-90° C., and the time of the heating reaction is 6-14 hours.

7. The preparation method according to claim 4, characterized in that: The structures of the tryptamine isonitrile derivatives are shown in formulas (1) to (4):

8. The preparation method according to claim 4, characterized in that: The structures of the unsaturated barbituric acid derivatives are shown in formulas (5)-(10):

9. Use of the spirocyclic barbituric acid derivative according to any one of claims 1 to 3 in the preparation of an anti-tumor drug, characterized in that: The types of tumors included breast cancer, human oral epidermoid carcinoma, and cervical cancer.

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