A small molecule inhibitor of TEAD and its use in TEAD receptor-related cancer drugs

Abstract

This invention relates to the field of medicinal chemistry, and particularly to a small molecule inhibitor of TEAD and its use in drugs for TEAD receptor-related cancers. Seven compounds were identified as TEAD inhibitors through virtual screening, and one compound was ultimately determined to have inhibitory effects on TEAD cells. This compound can therefore serve as a lead compound for the preparation of drugs for treating mesothelioma and other diseases related to the TEAD target. Specifically, the compound is: N-(4-fluorophenyl)-6-methyl-2-[5-methyl-1-(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide. This invention effectively screens ligands that inhibit the TEAD receptor using a combination of the AutoGen software for automated molecule generation and bioactivity assays. These ligands can serve as lead compounds for the preparation of drugs for treating TEAD receptor-mediated diseases, especially mesothelioma, and can effectively treat TEAD receptor-related diseases and their complications.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, in particular to a TEAD small molecule inhibitor and its use in TEAD receptor related cancer drugs. BACKGROUND

[0002] The Hippo signaling pathway was first discovered in Drosophila, and is a highly conserved signaling pathway across higher vertebrates, which regulates key target genes to control a variety of biological processes, including cell proliferation, survival, differentiation, cell fate determination, organ size and tissue homeostasis. The Hippo signaling pathway plays a very important role in inhibiting tumors. Since the Hippo signal is disordered, it will lead to the formation of tumors, so the Hippo signaling pathway is also one of the important anti-cancer targets.

[0003] The core of the Hippo pathway is composed of a series of kinase cascades. MST1 / 2 is a STE20 family protein kinase that can phosphorylate the adaptor protein SAV (Salvador homolog), the large tumor suppressor protein LAST1 / 2 and the MOB kinase activator. Activated LAST1 / 2 then phosphorylates YAP / TAZ, which is a transcriptional coactivator. Phosphorylated YAP / TAZ interacts with 14-3-3 protein and causes the complex to stay in the cytoplasm. In addition, the phosphorylation of YAP / TAZ can also lead to the degradation of YAP. When the Hippo pathway is inhibited, YAP / TAZ dephosphorylates and enters the nucleus, induces gene transcription by interacting with the transcription factor TEADs, and thus leads to the occurrence of tumors.

[0004] TEAD (transcriptional enhancer factor with TEA / ATTS domain) transcription factor is the final effector of the Hippo signaling pathway. The mammalian TEAD protein family contains four members, TEAD1-4, and the four TEADs have 61%-73% overall homology. YAP / TAZ and TEAD cannot start gene expression by themselves, only their complex can bind to DNA. Therefore, this provides a feasible strategy for designing targeted inhibitors. In addition, further studies have also found that targeting the TEAD palmitoylation hydrophobic pocket to regulate the Hippo pathway is a new strategy.

[0005] More and more studies have found that the TEAD receptor plays a very key role in many diseases, including mesothelioma, non-small cell carcinoma, liver cancer, etc. Therefore, the discovery of TEAD receptor inhibitors has become an important project of major pharmaceutical companies. SUMMARY

[0006] In view of the problems mentioned in the background art, the purpose of the present application is to provide a TEAD small molecule inhibitor and its use in a TEAD receptor related cancer drug; seven compounds discovered by virtual screening as TEAD inhibitors, 28 compounds purchased after virtual screening were subjected to activity determination, and finally one compound was determined to have the effect of inhibiting TEAD cells, so as to be used as a drug lead compound for preparing a drug for treating mesothelioma and other diseases related to the TEAD target.

[0007] The above technical purpose of the present application is achieved by the following technical solution: a TEAD small molecule inhibitor, specifically N-(4-fluorophenyl)-6-methyl-2-[5-methyl-1-(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide.

[0008] As an improvement, the screening is carried out by the following steps:

[0009] Step one, receptor preparation: the protein crystal structure with PDB library number 7CNL is extracted from the PDB library as the initial structure;

[0010] Step two, virtual screening: AutoGen is used as a tool to automatically generate potential active molecules;

[0011] Step three, biological test: find similar compounds to the potential active molecules generated by AutoGen in the MolPort compound molecular database, purchase and test.

[0012] The above technical purpose of the present application also provides a use of a TEAD small molecule inhibitor in a TEAD receptor related cancer drug, and the small molecule ligand of the TEAD receptor is the inhibitor of the TEAD receptor mentioned above.

[0013] As a preferred, the small molecule ligand of the TEAD receptor is a drug for treating TEAD receptor related diseases and their complications.

[0014] As a preferred, the TEAD receptor related disease can be mesothelioma, non-small cell carcinoma, liver cancer, ovarian cancer.

[0015] In summary, the present application mainly has the following beneficial effects: the present application can effectively screen out ligands for inhibiting TEAD receptors by combining the active molecule automatic generation software AutoGen and biological activity test, they can be used as drug lead compounds for preparing drugs for treating diseases related to the TEAD receptor, especially drugs for treating diseases such as mesothelioma, and can effectively treat TEAD receptor related diseases and be used as drugs for their complications. Attached Figure Description

[0016] Fig. 1 The molecular formula is N-(4-fluorophenyl)-6-methyl-2-[5-methyl-1-(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide;

[0017] Fig. 2 This is a schematic diagram of the analysis of N-(4-fluorophenyl)-6-methyl-2-[5-methyl-1-(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] like Figs. 1-2 As shown, the TEAD small molecule inhibitor of the present invention, after virtual screening, purchased 28 compounds and conducted activity assays, finally determined that one compound has the effect of inhibiting TEAD cells, and can therefore be used as a drug lead compound for the preparation of drugs for treating mesothelioma and other diseases related to the TEAD target, specifically: N-(4-fluorophenyl)-6-methyl-2-[5-methyl-1-(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide.

[0020] Small molecule ligands for the TEAD receptor are inhibitors of the aforementioned TEAD receptor; small molecule ligands for the TEAD receptor are drugs for treating TEAD receptor-related diseases and their complications; specific TEAD receptor-related diseases can include mesothelioma, non-small cell carcinoma, liver cancer, and ovarian cancer.

[0021] The discovery of the above compounds stemmed from the use of AutoGen, an automated active molecule generation software with proprietary intellectual property rights developed by Shanghai Aotu Zhiyao Biopharmaceutical Co., Ltd., which generated potential molecules with possible activity. Then, similar potential compounds with similar biological activities were searched in the MolPort (https: / / www.molport.com) compound library. A total of 28 compounds were purchased in this screening, of which 7 compounds had cellular-level biological activity of less than 10 μM, including the molecule in this patent.

[0022] The specific implementation method for compound generation and selection used in this invention is as follows:

[0023] Step 1: The protein crystal structure with PDB code 7CNL was extracted from the PDB library (Protein Data Bank) as the initial structure;

[0024] Step 2: Focusing on the ligands in the complex, The radius is defined as the pocket;

[0025] Step 3: Use AutoGen as a tool to automatically generate potentially active molecular compounds;

[0026] Step 4: Search the MolPort compound molecule database for compounds similar to the potentially active molecules generated by AutoGen, purchase them, and test them.

[0027] This invention, developed by Shanghai Aotu Zhiyao Biopharmaceutical Co., Ltd., combines the proprietary AutoGen software for automatically generating active molecules with a method for bioactivity testing to effectively screen ligands that inhibit TEAD receptors. These ligands can serve as lead compounds for the preparation of therapeutic drugs for TEAD receptor-mediated diseases, particularly mesothelioma.

[0028] Biological testing method: NCI-H226 cell inhibition assay

[0029] Dilute the compound in an LDV plate, repeatedly allocating 440 nL of the compound to the compound plate (Greiner-6008290), with a final DMSO concentration of 0.5%. Collect cells and resuspend them in growth medium to a density of 5E+4 cells / mL. Seed 100 μL of the appropriately concentrated cell suspension into the assay plate. Centrifuge the cell plate at 300 rpm for 1 minute and incubate the cells at 37°C, 95% humidity, and 5% CO2 for 24 hours. Add 40 μL of growth medium to the compound plate, then transfer 10 μL to the cell plate and centrifuge at 300 rpm for 1 minute. Incubate the cells at 37°C, 95% humidity, and 5% CO2 for 7 days. Equilibrate the plate and its compound at room temperature for approximately 15 minutes. Add 50 μL of CellTiter-Glo reagent to the assay plate; mix the compound on a oscillator for 20 minutes to induce cell lysis; incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal; read the plate using a reader equipped with luminescence readings. The experimental results are shown in Tables 1 and 2.

[0030] Table 1. Compound structures and corresponding bioactivity values

[0031]

[0032] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. Use of a TEAD small molecule inhibitor of N-(4-fluorophenyl)-6-methyl-2-[5-methyl- 1 -(4-methylphenyl)pyrazol-4-yl]quinoline-4-carboxamide in the preparation of a medicament for the treatment of a TEAD receptor-related cancer, characterized in that, The TEAD receptor related disease is specifically mesothelioma, non-small cell carcinoma, liver cancer, and ovarian cancer.

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