This review discusses recent advances in drug discovery related to β-lactam antibiotics, focusing on β-lactamases and cell wall transpeptidases. β-lactams are a class of antibiotics that inhibit penicillin-binding proteins (PBPs), which are involved in peptidoglycan synthesis. A major resistance mechanism involves β-lactamase enzymes that hydrolyze β-lactam antibiotics. Recent efforts have focused on developing novel inhibitors against β-lactamases, creating new β-lactams less susceptible to resistance, and identifying non-β-lactam inhibitors against transpeptidases. The review highlights the importance of understanding the targets, screening methods, and new inhibitor chemotypes in this field. It also discusses the development of cyclic boronate compounds that inhibit all classes of β-lactamases and the DBO series of small molecules that may lead to new antibiotics by targeting PBPs. The review emphasizes the challenges and opportunities in this area, including the need for new inhibitor chemotypes to counter future resistance. It also covers the development of new screening methods, such as microscale thermophoresis and surface plasmon resonance, and the use of computational methods in drug discovery. The review concludes with the development of new β-lactamase inhibitors and transpeptidase inhibitors, highlighting the potential for future drug discovery in this field.This review discusses recent advances in drug discovery related to β-lactam antibiotics, focusing on β-lactamases and cell wall transpeptidases. β-lactams are a class of antibiotics that inhibit penicillin-binding proteins (PBPs), which are involved in peptidoglycan synthesis. A major resistance mechanism involves β-lactamase enzymes that hydrolyze β-lactam antibiotics. Recent efforts have focused on developing novel inhibitors against β-lactamases, creating new β-lactams less susceptible to resistance, and identifying non-β-lactam inhibitors against transpeptidases. The review highlights the importance of understanding the targets, screening methods, and new inhibitor chemotypes in this field. It also discusses the development of cyclic boronate compounds that inhibit all classes of β-lactamases and the DBO series of small molecules that may lead to new antibiotics by targeting PBPs. The review emphasizes the challenges and opportunities in this area, including the need for new inhibitor chemotypes to counter future resistance. It also covers the development of new screening methods, such as microscale thermophoresis and surface plasmon resonance, and the use of computational methods in drug discovery. The review concludes with the development of new β-lactamase inhibitors and transpeptidase inhibitors, highlighting the potential for future drug discovery in this field.