Application of pyrazolo pyrazinone compound

Abstract

The invention discloses an application of a pyrazolo pyrazinone compound. According to the application of the pyrazolo pyrazinone compound in preparation of drugs for preventing and / or treating cancers, the structure of the pyrazolo pyrazinone compound is shown in the specification. The inventor of the invention finds that the pyrazolo pyrazinone compound has excellent activity on various different types of human tumors for the first time by performing activity screening on a small molecular skeleton, and provides an optional scheme for broad-spectrum anti-cancer drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of a pyrazolopyrazinone compound. Background Technology

[0002] Cancer is currently one of the leading causes of death worldwide, posing a serious threat to human health. Over the past three decades, significant progress has been made in the research and development of anti-tumor drugs. Treatment strategies have gradually shifted from traditional chemotherapy to molecularly targeted therapy and immunotherapy, forming a new treatment landscape with multiple mechanisms. Statistics show that small molecule drugs account for approximately 78% of the 174 widely used anti-tumor drugs globally. Therefore, developing a product that can solve the aforementioned technical challenges is essential. Summary of the Invention

[0003] The purpose of this invention is to provide an application of a pyrazolopyrazinone compound.

[0004] The object of this invention is achieved by the use of the pyrazolopyrazinone compound in the preparation of drugs for the prevention and / or treatment of cancer, wherein the structure of the pyrazolopyrazinone compound is as follows: .

[0005] The pyrazolopyrazinone compound contains the pyrazolopyrazinone heterocyclic system (pyrazolo[1,5-a]pyrazin-4(5H)-one), which is a common kinase inhibitor skeleton that can mimic ATP binding to the target. At the 2-position of the core, a benzodioxanepentenyl group (i.e., the common "piper ring" structure, a benzene ring fused with a five-membered dioxane ring) is directly attached. This group has unique electronic properties and steric hindrance. These structures integrate multiple pharmacophores commonly found in anticancer drugs. Although studies have reported the wide application of this type of compound in the pharmaceutical field, its comprehensive chemical composition and antitumor mechanism are still relatively few reported.

[0006] The pyrazolopyrazinone compound 2-(benzo[d][1,3]dioxol-5-yl)-5-[(5-methyl-2-(3,4,5-trimethoxyphenyl)oxazol-4-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-one described in this invention has the English name: (2-(benzo[d][1,3]dioxol-5-yl)-5-((5-methyl-2-(3,4,5-trimethoxyphenyl)oxazol-4-yl)methyl)pyrazolo[1,5-a]pyrazin-4(5H)-one H )-one) This invention has revealed that (2-(benzo[d][1,3]dioxacyclopenten-5-yl)-5-[(5-methyl-2-(3,4,5-trimethoxyphenyl)oxazol-4-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-one exhibits strong inhibitory activity against tumor cells. Based on the cell activity experiments of this invention, the pyrazolopyrazinone compound described herein showed clear and differential activity against human breast cancer cells, human liver cancer cells, human lung cancer cells, and human osteosarcoma cells in vitro.

[0007] The inventors of this invention, through activity screening of small molecule skeletons, discovered for the first time that the pyrazolopyrazinone compound exhibits excellent activity against a variety of different types of human tumors, providing an option for broad-spectrum anticancer drugs. Attached Figure Description

[0008] Figure 1 The graph shows the inhibition curves of the pyrazolopyrazinone compound on the viability of five types of human tumor cells. Detailed Implementation

[0009] The present invention will be further described below with reference to embodiments, but this is not intended to limit the present invention in any way. Any modifications or substitutions made based on the teachings of the present invention shall fall within the protection scope of the present invention.

[0010] The application of the pyrazolopyrazinone compound of the present invention in the preparation of drugs for the prevention and / or treatment of cancer, wherein the structure of the pyrazolopyrazinone compound is as follows: It originates from a drug-like compound library containing 5000 compounds developed by Shanghai Taoshu Biotechnology Co., Ltd. The pyrazolopyrazinone compound has the Library ID G825-0328, Plate layout PHD198946, CAS Number 1287101-04-5, and chemical formula C. 27 H 24 N4O7 has a relative molecular mass of 516.51. The pyrazolopyrazinone compounds used in Examples 1-4 were purchased from Shanghai Taoshu Biotechnology Co., Ltd.

[0011] The cancers mentioned are breast cancer, liver cancer, colorectal cancer, bone cancer, or lung cancer.

[0012] The human breast cancer cells used in the following examples are HCC1806, the human liver cancer cells are human liver cancer cells LM3, the human lung cancer cells are human lung cancer cells A549, the human osteosarcoma cells are human osteosarcoma cells MG63, and the human colorectal cancer cells are human ileocecal colorectal adenocarcinoma cells HCT-8. All the conventional cancer cells used above were obtained from Kunming Medical University.

[0013] Unless otherwise specified, all reagents used in the embodiments of this invention are commercially available. RPMI 1640 medium, DMEM medium, and fetal bovine serum were purchased from Biological Industries; phosphate-buffered saline (PBS) and 0.25% trypsin (containing EDTA) were purchased from Gibco.

[0014] Cell lines were cultured in DMEM or RPMI 1640 medium containing 10% fetal bovine serum at 37°C, 5% CO2 and 90% humidity.

[0015] Example 1

[0016] The CCK-8 assay was used to detect cell growth inhibition. The CCK-8 assay, short for Cell Counting Kit-8, is a commonly used method for detecting cell proliferation and cytotoxicity. The principle of the CCK-8 assay is based on the compound WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid phenyl)-2H-tetrazole monosodium salt). In the presence of the electron carrier 1-Methoxy PMS (menaquinone phosphate), WST-8 can be reduced by intracellular mitochondrial dehydrogenases to produce a highly water-soluble orange-yellow formazan product. The amount of formazan produced is directly proportional to the number of viable cells, and the color intensity reflects cellular metabolic activity. The number of viable cells can be indirectly reflected by measuring the absorbance at 450 nm using a microplate reader. Therefore, CCK-8 can be used to assess cell proliferation, cytotoxicity, or the cellular effects of drugs.

[0017] (2-(benzo[d][1,3]dioxacyclopenten-5-yl)-5-[(5-methyl-2-(3,4,5-trimethoxyphenyl)oxazol-4-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-one was dissolved in DMSO (dimethyl sulfoxide), and then solutions with DMSO were prepared at concentrations of 10 μM, 5 μM, 1 μM, 0.5 μM and 0.1 μM, respectively. These solutions were used as the test solutions.

[0018] Human breast cancer cells HCC1806 (4×10) 3Cells were seeded in 96-well plates with 99 μL of culture medium per well and incubated at 37°C with 5% CO2 for 24 h. Then, 1 μL of the test solution was added to each well. For the control group, only 1 μL of DMSO was added to the cell culture, and the plates were incubated for 72 h. Then, 10 μL of CCK8 solution was added to each well, and the mixture was gently mixed to avoid air bubbles. The plates were incubated for another 1 h, and the absorbance of each well was measured at 450 nm using a microplate reader. The average OD value of each well was calculated, and the results were compared with the control group (untreated cells) to analyze changes in cell proliferation or toxicity. The percentage of the absorbance value of the experimental group relative to the absorbance value of the control group represents the cell viability or proliferation level; the control group is assumed to be 100%.

[0019] The results are shown in Table 1 and Figure 1 As shown: Table 1. Inhibitory effects of pyrazolopyrazinone compounds on human breast cancer cell line HCC1806.

[0020] According to Table 1 and Figure 1 The results showed that the pyrazolopyrazinone compound (CAS No. 1287101-04-5) exhibited cancer cell toxicity against the human breast cancer cell line HCC1806.

[0021] Example 2

[0022] The human breast cancer cell line HCC1806 from Example 1 was replaced with the human osteosarcoma cell line MG63. Other procedures were the same as in Example 1. Results are shown in Table 2 and... Figure 1 As shown.

[0023] Table 2. Inhibitory effects of pyrazolopyrazinone compounds on human osteosarcoma cell line MG63.

[0024] According to Table 2 and Figure 1 The results showed that the pyrazolopyrazinone compound (CAS No. 1287101-04-5) exhibited cancer cell toxicity against the human osteosarcoma cell line MG63.

[0025] Example 3

[0026] The human breast cancer cell line HCC1806 was replaced with the human liver cancer cell line LM3. Other procedures were the same as in Example 1. Results are shown in Table 3. Figure 1 As shown.

[0027] Table 3. Inhibitory effects of pyrazolopyrazinone compounds on human hepatocellular carcinoma cell line LM3.

[0028] According to Table 3 and Figure 1 The results showed that the pyrazolopyrazinone compound (CAS No. 1287101-04-5) exhibited cancer cell toxicity against the human liver cancer cell line LM3.

[0029] Example 4

[0030] The human breast cancer cell line HCC1806 was replaced with human lung cancer cell line A549. Other procedures were the same as in Example 1. Results are shown in Table 4. Figure 1 As shown.

[0031] Table 4. Inhibitory effects of pyrazolopyrazinone compounds on human lung cancer cell line A549.

[0032] According to Table 4 and Figure 1 The results showed that the pyrazolopyrazinone compound (CAS No. 1287101-04-5) exhibited cancer cell toxicity against the human lung cancer cell line A549.

[0033] Example 5

[0034] The human breast cancer cell line HCC1806 was replaced with human ileocecal colorectal adenocarcinoma cell line HCT-8. Other procedures were the same as in Example 1, and the results are shown in Tables 5 and 6. Figure 1 As shown.

[0035] Table 5. Inhibitory effects of pyrazolopyrazinone compounds on human ileocecal colorectal adenocarcinoma cell line HCT-8.

[0036] According to Table 5 and Figure 1 The results showed that the pyrazolopyrazinone compound (CAS No. 1287101-04-5) exhibited cancer cell toxicity against the human ileocecal colorectal adenocarcinoma cell line HCT-8.

[0037] In summary: as shown in Tables 1-5 and Figure 1 The compound (2-(benzo[d][1,3]dioxacyclopenten-5-yl)-5-[(5-methyl-2-(3,4,5-trimethoxyphenyl)oxazol-4-yl)methyl]pyrazolo[1,5-a]pyrazin-4(5H)-one showed significant dose-dependent inhibitory activity against human breast cancer cells, human osteosarcoma cells, human liver cancer cells, human lung cancer cells, and human ileocecal colorectal adenocarcinoma cells. Cell activity decreased with increasing drug concentration, indicating that this compound has a clear anti-proliferative effect.

[0038] The above are merely embodiments of the present invention and do not limit the scope of the patent. Any equivalent modifications made based on the content of this specification, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of the present invention.

Claims

1. The application of a pyrazolopyrazinone compound, characterized in that, The pyrazolopyrazinone compound is used for preparing a medicine for preventing and / or treating cancer, and the structure of the pyrazolopyrazinone compound is as follows: 。 2. Use according to claim 1, characterized in that, The cancer is breast cancer, liver cancer, intestinal cancer, bone cancer or lung cancer.

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