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US-20260125350-A1 - CENTROMERE PROTEIN CENP-M SMALL MOLECULE INHIBITOR CENPEMLIN, AND PREPARATION METHOD THEREFOR AND USE THEREOF

US20260125350A1US 20260125350 A1US20260125350 A1US 20260125350A1US-20260125350-A1

Abstract

Provided are a centromere protein CENP-M small molecule inhibitor cenpemlin, and a preparation method therefor and the use thereof. A small molecule compound cenpemlin is obtained by means of analyzing the protein plane information of the fine structures of CENP-M and CENP-L and performing targeted screening. The results of biochemical and cytological experiments all indicate that cenpemlin inhibits the proliferation of cancer cells by means of interfering with the interaction between CENP-M and CENP-L, thereby providing a new compound for analyzing and interfering with the rapid proliferation of cancer cells.

Inventors

  • Xuebiao YAO
  • Xing Liu
  • Fengrui YANG
  • Mingming Ma

Assignees

  • UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA

Dates

Publication Date
20260507
Application Date
20220727
Priority Date
20220711

Claims (15)

  1. 1 . A compound of formula I, isomers thereof, and pharmaceutically acceptable salts thereof:
  2. 2 . A method for preparing the compound of formula I according to claim 1 , comprising the following steps: 1) reacting cyanuric chloride and morpholine under an alkaline condition to generate a cyanuric chloride intermediate 1 containing a di-1,4-oxo-nitrogen heterocycle; 2) subjecting the cyanuric chloride intermediate 1 containing the di-1,4-oxo-nitrogen heterocycle to a substitution reaction with hydrazine hydrate to generate compound 2, wherein the compound 2 is 1,3(1,4-oxo-nitrogen heterocycle)-5-hydrazine cyanuric acid; 3) subjecting the 1,3(1,4-oxo-nitrogen heterocycle)-5-hydrazine cyanuric acid to a condensation reaction with a hexacarbon monosaccharide in glacial acetic acid to obtain the compound of formula I.
  3. 3 . The method according to claim 2 , wherein in the step 1), a molar ratio of the cyanuric chloride to the morpholine is 1: 2-2.5; the reaction is carried out under an action of an ice-water bath, and a reaction time is 1-1.5 h; the alkaline condition is provided by sodium bicarbonate.
  4. 4 . The method according to claim 2 , wherein in the step 2), a molar ratio of the cyanuric chloride intermediate 1 containing the di-1,4-oxo-nitrogen heterocycle to the hydrazine hydrate is 1: 1-2; a reaction temperature is 40° C.-60° C., a reaction time is 3-4 h.
  5. 5 . The method according to claim 2 , wherein in the step 3), a ratio of the 1,3(1,4-oxo-nitrogen heterocycle)-5-hydrazine cyanuric acid to the hexacarbon monosaccharide is 10 mmol: 1.8-2.0 g; a reaction temperature is 40° C.-60° C., a reaction time is 4-5 h.
  6. 6 . A method for preparing a product, comprising using the compound of formula I according to claim 1 , the isomers thereof, and the pharmaceutically acceptable salts thereof, wherein the product comprises: 1) an inhibitor of centromere protein CENP-M; 2) an agent for inhibiting cell mitosis; 3) a product for inhibiting tumor cell proliferation; and 4) a product for preventing and/or treating a cancer.
  7. 7 . The method according to claim 6 , wherein inhibiting cell mitosis is achieved by interfering with CENP-M-mediated kinetochore assembly.
  8. 8 . The method according to claim 6 , wherein tumor cells are epithelial cancer cells, and the epithelial cancer cells comprise cervical cancer cells, liver cancer cells, and breast cancer cells.
  9. 9 . The method according to claim 9 , which wherein the cancer is epithelial cancer, and the epithelial cancer comprises cervical cancer, liver cancer, and breast cancer.
  10. 10 . A reagent for inhibiting cell mitosis, comprising the compound of formula I according to claim 1 , the isomers thereof, and the pharmaceutically acceptable salts thereof.
  11. 11 . A product for inhibiting tumor cell proliferation, comprising the compound of formula I according to claim 1 , the isomers thereof, and the pharmaceutically acceptable salts thereof.
  12. 12 . A product for preventing and/or treating cancer, comprising the compound of formula I according to claim 1 , the isomers thereof, and the pharmaceutically acceptable salts thereof.
  13. 13 . A method for inhibiting an interaction between CENP-M and CENP-L, comprising: adding the compound of formula I according to claim 1 , the isomers thereof, and the pharmaceutically acceptable salts thereof to a system containing the CENP-M and the CENP-L, and incubating the system.
  14. 14 . The method according to claim 3 , wherein in the step 2), a molar ratio of the cyanuric chloride intermediate 1 containing the di-1,4-oxo-nitrogen heterocycle to the hydrazine hydrate is 1: 1-2; a reaction temperature is 40° C.-60° C., a reaction time is 3-4 h.
  15. 15 . The method according to claim 3 , wherein in the step 3), a ratio of the 1,3(1,4-oxo-nitrogen heterocycle)-5-hydrazine cyanuric acid to the hexacarbon monosaccharide is 10 mmol: 1.8-2.0 g; a reaction temperature is 40° C.-60° C., a reaction time is 4-5 h.

Description

CROSS REFERENCE TO THE RELATED APPLICATIONS This application is the national phase entry of International Application No. PCT/CN2022/108196, filed on Jul. 27, 2022, which is based upon and claims priority to Chinese Patent Application No. 202210809550.2, filed on Jul. 11, 2022, the entire contents of which are incorporated herein by reference. TECHNICAL FIELD The present invention relates to the field of medicine, and specifically relates to cenpemlin, a small molecule inhibitor of CENP-M, a key centromere protein of mitosis, and a preparation method and use thereof. BACKGROUND Precise cell mitosis ensures the stability of the genome and the maintenance of cell homeostasis. Mitosis produces two genetically identical daughter cells through the equal separation of sister chromatids. The molecular mechanism driving chromosome separation is relatively conserved during the evolution of eukaryotes. The kinetochore is an essential element of this molecular mechanism and can be regarded as a layered structure, with its inner and outer layers directly contacting the centromere chromatin and spindle microtubules, respectively. The inner layer of the kinetochore is composed of at least 16 centromere proteins, which assemble into multi-complexes and directly interact with the centromere chromatin. These complexes include: CENP-H, I, K, M complex, CENP-O, P, Q, U, R complex, CENP-T, W, S, X complex, CENP-L, N complex and CENP-C protein. After these complexes are assembled, they further recruit the remaining kinetochore proteins. Although the interactions between centromere proteins have been widely studied, the specific biological functions of some of these proteins still need to be further explored. Recently, our group used cryo-electron microscopy to analyze the structure of the CCAN complex. The CCAN complex is in the shape of Chinese character “human”, with CENP-L/N dimers forming an arch, and DNA passing through the bottom of the arch. The CENP-M protein interacts with the other three centromere proteins CENP-H, I, and K to form a complex and become part of the inner layer of the kinetochore. CENP-M binds near the CENP-L/N dimer at the top of the arch, and has an interaction interface with both CENP-L and CENP-N in space. At the same time, CENP-M is localized at the kinetochore throughout the cell cycle, and its function may be constitutive, but the specific molecular mechanism has not yet been fully resolved. CENP-M was originally named PANE1 (proliferation associated nuclear element 1) and is only conserved in metazoans. Subsequent studies have shown that CENP-M is closely related to CENP-A, CENP-L, CENP-N, and CENP-T. The loss of CENP-M leads to the mislocalization of other CCAN proteins, indicating that CENP-M is essential for the assembly and stability of the inner kinetochore. Although CENP-M is structurally and evolutionarily related to GTPase, but it is not a true enzyme. By forming a quaternary complex with the evolutionarily conserved CENP-H, I, and K proteins, it plays a vital role in a series of important biological processes such as recruiting the chromatin remodeling FACT protein complex, promoting CENP-A nucleosome remodeling, and recruiting spindle checkpoint-related proteins. There are currently no reports on CENP-M inhibitors. SUMMARY The purpose of the present invention is to provide a compound, an isomer thereof, and a pharmaceutically acceptable salt thereof, and a preparation method and use thereof. The compound provided by the present invention has a structural formula as shown in Formula I: The compound shown in the above Formula I specifically targets the key centromere protein CENP-M in mitosis, and is a CENP-M inhibitor designated as cenpemlin. CENP-M is an important member of the inner kinetochore CCAN complex. It was named PANE1 because it was identified in rapidly proliferating cells. Subsequent studies found that CENP-M is a pseudo-GTPase. In recent years, studies on the inner kinetochore CCAN complex have emerged in an endless stream, but its fine structure has not been fully resolved. We use structural biology methods based on cryo-electron microscopy to answer scientific questions. The CCAN complex proteins were expressed and purified using an insect system, and then recombined in vitro to obtain a complete CCAN complex. The three-dimensional image of the complex was collected by cryo-electron microscopy, and then three-dimensional reconstruction was performed. Finally, we obtained the structural information of the CCAN complex containing the CENP-M protein. Based on the fine structure of the interaction interface between CENP-M and CENP-L proteins, we identified a specific CENP-M targeting small molecule compound, cenpemlin, by combining molecular docking analysis, virtual screening, cell phenotype analysis and protein interaction experiments. The compound shown in Formula I was prepared by a method including the following steps according to the synthetic route shown in FIG. 1: 1)