Abstract
Human epidermal growth factor receptor 2-positive (HER2+) breast cancer is correlated with poor prognosis, the current treatment of which is still based on surgery and adjuvant targeted therapy with monoclonal antibody. Problems of drug resistance hinder the use of monoclonal antibodies. Subsequently, tyrosine kinase inhibitors (TKIs) have been noticed, TKIs have the advantages of multi-targets and reduced drug resistance. However, TKIs that target HER family proteins often cause adverse effects such as liver damage and diarrhea. Thus, TKIs with high selectivity are being developed. TH-4000, a prodrug that generated an active form TH-4000Effector (TH-4000E) under hypoxic condition, was evaluated in this research. We found that TH-4000E ([(E)-4-[[4-(3-bromo-4-chloroanilino)pyrido[3,4-d]pyrimidin-6-yl]amino]-4-oxobut-2-enyl]-dimethyl-[(3-methyl-5-nitroimidazol-4-yl)methyl]azanium) (1–1000 nM) had potent and highly selective toxic effects on HER2+ breast cancer cells and inhibited the phosphorylation of HER family kinases at lower doses than that of Lapatinib and Tucatinib. TH-4000E activated Caspase-3 and induced apoptosis through a reactive oxygen species (ROS)-dependent pathway. The prodrug TH-4000 ([(E)-4-[[4-(3-bromo-4-chloroanilino)pyrido[3,4-d]pyrimidin-6-yl]amino]-4-oxobut-2-enyl]-dimethyl-[(3-methyl-5-nitroimidazol-4-yl)methyl]azanium;bromide) (50 mg/kg) effectively suppressed the tumor growth with less liver damage in mouse tumor models. This hypoxia-targeted strategy has possessed advantage in avoiding drug-induced liver damage, TH-4000 could be a promising drug candidate for the treatment of HER2+ breast cancer.
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Data availability
The data is available upon request from the the lead contact, Dawei Yang (David_young1994@outlook.com).
Abbreviations
- EGFR:
-
Human epidermal growth factor receptor
- HER2:
-
Human epidermal growth factor receptor 2
- HER3:
-
Human epidermal growth factor receptor 3
- HER4:
-
Human epidermal growth factor receptor 4
- ERK:
-
Extracellular regulated protein kinase
- AKT:
-
Protein kinase b
- OS:
-
Overall survival
- PFS:
-
Progression-free survival
- ATP:
-
Adenosine triphosphate
- FDA:
-
Food and drug administration
- TKI:
-
Tyrosine kinase inhibitor
- TH-4000E:
-
TH-4000Effector
- DMSO:
-
Dimethyl sulfoxide
- CCK-8:
-
Cell counting kit-8
- IC50:
-
Half inhibitory concentration
- FBS:
-
Fetal bovine serum
- PBS:
-
Phosphate buffered saline
- DMEM:
-
Dulbecco’s modified eagle’s medium
- ROS:
-
Reactive oxygen species
- NAC:
-
N-Acetyl-l-cysteine
- Z-VAD:
-
Z-VAD(OMe)-FMK
- PARP:
-
Poly adp-ribose polymerase
- MMP:
-
Mitochondrial membrane potential
- CD36:
-
Platelet glycoprotein 4
- CPT1A:
-
Carnitine palmitoyltransferase 1A
- CPT1B:
-
Carnitine palmitoyltransferase 1B
- FAO:
-
Fatty acid oxidation
- mRNA:
-
Messenger RNA
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- qPCR:
-
Real-time quantitative PCR
- PI:
-
Propidium iodide
- IHC:
-
Immunohistochemistry
- HE:
-
Hematoxylin–eosin
- ALT:
-
Alanine aminotransferase
- AST:
-
Aspartate aminotransferase
- CREA:
-
Creatinine
- MRM:
-
Multiple reaction monitoring
- NSCLC:
-
Non-small cell lung cancer
- DLT:
-
Dose limiting toxicity
- MTD:
-
Maximum tolerated dose
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Acknowledgements
This research was supported by the Natural Science Foundation of Jiangsu province (No. BK20230157), the Medical Engineering Cooperation Project of Nanjing Municipal Science and Technology Bureau (No. 202110032), the Talent Support Program by the Second Hospital of Nanjing (No. RCMS23004), the High-level Talents Program from the Second Hospital of Nanjing (No. 202302), Nanjing Pukou Program for Society and Science Development (No. S2022-1), Nanjing Healthcare and Science Development Program (No. YKK22231), Guangdong Basic and Applied Basic Research Foundation (2019A1515110993) and the Postgraduate Research and Practice Innovation Program of Jiangsu Province (No. KYCX22_1927).
We thank the Center for Analysis and Testing of China Pharmaceutical University for the use of HPLC‐QQQ‐MS/MS, and would like to thank Zi-Yuan Wang (China Pharmaceutical University).
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Shao, X., Yang, D., Shan, L. et al. TH-4000, a hypoxia-activated pan-HER inhibitor, shows excellent preclinical efficacy for the treatment of HER2+ breast cancer. Arch Toxicol 98, 865–881 (2024). https://doi.org/10.1007/s00204-023-03670-6
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DOI: https://doi.org/10.1007/s00204-023-03670-6