Network Pharmacology and In Vitro Cell Experimental Study for Exploring the Therapeutic Potential of Thiazolone Derivatives in Rhabdomyosarcoma

Summary

We present a study that systematically investigates the therapeutic potential of thiazolone derivatives against rhabdomyosarcoma based on network pharmacology and computational chemistry, with validation and evaluation performed through in vitro cell experiments.

Abstract

Rhabdomyosarcoma (RD) is a malignant tumor originating from rhabdomyoblasts, which poses a serious threat to the health of children and adolescents. Based on the potential of thiazolone derivatives in a variety of biological activities, this study intends to systematically explore the mechanisms of thiazolone derivatives in the treatment of RD by combining network pharmacology and in vitro cell experiments. Through network pharmacology, high-throughput data were integrated to predict the potential targets of thiazolone. Additionally, AutoDock Vina software was used to calculate the binding energy of the compound and the target protein molecule, revealing its signaling pathway related to RD. Finally, in vitro cell experiments further verified the tumor suppressive effect of the compounds. Network pharmacology was used to analyze the targets of thiazolone derivatives in the treatment of RD, and 172 target genes were finally obtained by combining and deduplicating. Five protein targets with low binding energy were screened by docking between the compound and the target protein molecule, PIK3CD, AKT1, GSK3B, HSP90AB1, and ITGB1. The results of in vitro cell culture experiments showed that a specific thiazolone derivative had a significant inhibitory effect on RD cell lines, providing a scientific basis for the discovery of new targets and effective compounds for the treatment of RD. This study revealed the mechanisms of thiazolone derivatives in the treatment of RD through a combination of network pharmacology and in vitro experiments and provided a new perspective for future drug development and treatment strategies.

Introduction

Rhabdomyosarcoma (RD), a highly aggressive pediatric soft tissue sarcoma that represents a significant threat to pediatric health, with limited therapeutic options. Epidemiological studies indicate that while RD exhibits relatively low incidence rates, its high malignancy and poor prognosis present substantial clinical challenges. RD is classified into four distinct histological subtypes: embryonal, alveolar, spindle cell/sclerosing, and pleomorphic, with embryonal rhabdomyosarcoma (ERMS) being the most prevalent form^1,2,3. Current therapeutic strategies for newly diagnosed l....

Protocol

All the network pharmacology procedures were carried out in accordance with the Guidelines for Network Pharmacology Evaluation Methods²¹. The comprehensive network pharmacology workflow adopted in this study is schematically illustrated in Figure 2.

Synthesis of Thiazolone derivatives
The thiazolone derivatives in this study were synthesized in our previous work¹⁹, through a copper-catalyzed remote asymmetric propargylation reaction. The reaction utilized CuI as a catalyst and a chiral Pybox ligand (L1) as the stereocontrol core, in a DIPEA ....

Results

In this study, 13 thiazolone derivatives (compounds a-m; Figure 3, Figure 4) were previously synthesized¹⁹ were selected for evaluation of their inhibitory activity against cancer cells (Figure 5). Among these compounds, compound m at 100 µM showed the highest inhibition rate against RD cells (Embryonal Rhabdomyosarcoma Cell) (STR Authenticated)²². Subsequent experiments involving.......

Discussion

Network pharmacology is commonly used to predict and screen potential active molecules within complex traditional Chinese medicine components, and to elucidate the synergistic mechanisms of multi-component, multi-target, and multi-pathway actions in herbal formulations. In this study, we applied a network pharmacology approach to systematically investigate the potential mechanism of our newly synthesized thiazolone derivative against RD by constructing a compound-target-pathway-disease network. In this study, we systemat.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
CCK-8 Assay Kit	Beyotime Biotechnology Co., Ltd.	C0038	A kit used for measuring cell viability and proliferation based on the reduction of a water-soluble tetrazolium salt to form a colored product.
Clean Bench	Suzhou Antai Airtech Co., Ltd	SW-CJ-1FD	A laminar airflow workstation that provides a sterile environment for handling cell cultures and other sensitive materials.
CO2 Incubator	Heal Force	HF90	A temperature, humidity, and CO2-controlled environment is used for cell culture to maintain optimal growth conditions.
DMEM High Glucose	cytiva	SH30022.01	A cell culture medium with high glucose concentration (4.5 g/L), suitable for a variety of cell types, including fast-growing mammalian cells.
FBS	YEASEN	40130ES76	A common supplement in cell culture media, providing essential growth factors, hormones, and nutrients for cell proliferation.
Inverted Microscope	OLYMPUS	IX51	A microscope designed with the objective lens below the stage, allowing observation of cell cultures in flasks or plates from below.
Microplate Reader	Diatek	DR-200Bs	An instrument used to detect biological, chemical, or physical events in microplate samples by measuring absorbance, fluorescence, or luminescence.
PBS	Jinuo Biomedical Technology Co., Ltd.	GNM20012	A buffer solution is commonly used in biological research for washing cells, diluting solutions, and maintaining pH stability.
RD (Embryonal Rhabdomyosarcoma Cell) (STR Authenticated)	Shanghai FuHeng Biotechnology Co., Ltd.	FH0712	The cell line has been authenticated by short tandem repeat (STR) analysis, possesses a well-defined genetic background and clear cell identity, thereby ensuring the reliability and reproducibility of experimental data.
RMPI Medium Modified (1640)	cytiva	SH30809.01	A widely used cell culture medium originally developed for human leukocyte cells, is now applied for various mammalian cell types.
Trypsin-EDTA	Jinuo Biomedical Technology Co., Ltd.	GNM25200	A solution used to detach adherent cells from culture surfaces by digesting extracellular proteins and chelating calcium ions.