A Review of Small Molecule Inhibitors and Functional Probes of Human Cathepsin L

Abstract

Human cathepsin L belongs to the cathepsin family of proteolytic enzymes with primarily an endopeptidase activity. Although its primary functions were originally thought to be only of a housekeeping enzyme that degraded intracellular and endocytosed proteins in lysosome, numerous recent studies suggest that it plays many critical and specific roles in diverse cellular settings. Not surprisingly, the dysregulated function of cathepsin L has manifested itself in several human diseases, making it an attractive target for drug development. Unfortunately, several redundant and isoform-specific functions have recently emerged, adding complexities to the drug discovery process. To address this, a series of chemical biology tools have been developed that helped define cathepsin L biology with exquisite precision in specific cellular contexts. This review elaborates on the recently developed small molecule inhibitors and probes of human cathepsin L, outlining their mechanisms of action, and describing their potential utilities in dissecting unknown function.Molecules 2020, 25, 698 2 of 41 also now well established and has been a subject of elegant reviews elsewhere [25][26][27]. Unfortunately, several overlapping and redundant functions of cathepsin L have also emerged [5,[28][29][30]. It is therefore critically important that its functional biology in both normal and disease-specific cell types be clearly annotated before significant resources are directed in drug discovery endeavors. In this review, we will briefly outline the biogenesis of cathepsin L, describe its post-translational processing and trafficking, and highlight key structural features required for formation of an active and mature cathepsin L. A brief summary of cathepsin L biology and its role in human diseases is provided next. Finally, a detailed and up-to-date report on existing cathepsin L-targeting small molecule inhibitors and functional probes with their mechanistic details is described.Human cathepsin L gene, CTHL (Uniprot primary accession number: P07711), located at the 9q21-q22 position of chromosome 9, encodes for a total of 333 amino acid peptide sequence (M.W. = 37,564 kDa) [31]. The coding space includes the regions of a N-terminal signal peptide, two pro-peptides, and two mature peptides comprising of a heavy (H) chain and a light (L) chain ( Figure 1). After transcription, the signal peptide is co-translationally removed in polysome and the resulting 41 kDa pro-cathepsin L is translocated to endoplasmic reticulum where it undergoes N-linked glycosylation with mannose rich sugars. The mannose sugars on the pro-cathepsin L are then phosphorylated in cis Golgi by UDP-N-acetylglucosamine:N-acetylglucosaminephosphotransferase enzyme [32]. The mannose-6-phosphate (M6P) receptors located on the surface of the trans Golgi network recognize the M6P-pro-cathepsin peptide and deliver the pro-cathepsin L peptide to the lysosome via the endolysosomal pathway. The weakly acidic environment of endosome/lysosome releases M6P receptors and the phosphate ...

Cathepsins in the Pathophysiology of Mucopolysaccharidoses: New Perspectives for Therapy

“…The inhibition of CTSs has been widely explored over the last decades in the field of chronic inflammatory diseases [27,37,204,205], cardiovascular diseases [10,19,181], osteoporosis [70][71][72][73], arthritis [28,206], kidney diseases [30][31][32]84], pancreatitis [207], obesity [208][209][210], cancer [25,34,48,74,82,211], neurodegenerative diseases [39,41,184,185,212,213], and many other pathological states. Multiple inhibitors are currently available, ranging from reversible covalent inhibitors to irreversible inhibitors [214][215][216][217][218] (Table 4). The class of reversible inhibitors acts by engaging the target protein through non-covalent interactions, whereas irreversible inhibitors bind the target protein with stable covalent bonds [219].…”

“…Moreover, off-target effects with broad-spectrum inhibition of proteases, leading to unpredictable side effects in clinical trials, has represented the main concern for the therapeutic use of CTS inhibitors in humans. The recent focus on target-and ligand-binding drug design to selectively inhibit specific CTSs has provided excellent results in overcoming such issue [19,72,[204][205][206][214][215][216][217][218][219][220][221][222]. In this context, several new strategies have been reported for successfully CTS targeting, including designed ankyrin repeat proteins (DARPins) with high CTSB blocking activity [222], non-peptide synthetic molecules with anti-CTSK [71] and anti-CTSD [223] activity, and naturally occurring asperphenamate [224].…”

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“…Based on the structure of E-64, the cathepsin L–specific inhibitor CLIK-148 was developed [ 167 ]. In addition to epoxysuccinyl inhibitors, irreversible cysteine peptidase inhibitors include epoxides, vinyl sulphones, aziridines, azomethylketones, halomethylketones, acyloxymethylketones, azapeptides, thiadiazoles, hydroxametes, β-lactams, and α,β-unsaturated carbonyl derivatives and others (reviewed in [ 152 , 154 ]).…”

Advances in Anti-Cancer Immunotherapy: Car-T Cell, Checkpoint Inhibitors, Dendritic Cell Vaccines, and Oncolytic Viruses, and Emerging Cellular and Molecular Targets

“…Cathepsins are lysosomal enzymes which are highly expressed in tumour cells in response to the hypoxic and slightly acid microenvironment. A number of these cathepsin enzymes has been implicated in the progression of tumour growth and metastasis in the last two decades [211][212][213]. Notably, aspartyl cathepsin D gained an increased attention due to their extracellular presence in the TME [211,214].…”