Proteases as therapeutics

Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA 94131, USA.
Biochemical Journal (Impact Factor: 4.78). 04/2011; 435(1):1-16. DOI: 10.1042/BJ20100965
Source: PubMed

ABSTRACT Proteases are an expanding class of drugs that hold great promise. The U.S. FDA (Food and Drug Administration) has approved 12 protease therapies, and a number of next generation or completely new proteases are in clinical development. Although they are a well-recognized class of targets for inhibitors, proteases themselves have not typically been considered as a drug class despite their application in the clinic over the last several decades; initially as plasma fractions and later as purified products. Although the predominant use of proteases has been in treating cardiovascular disease, they are also emerging as useful agents in the treatment of sepsis, digestive disorders, inflammation, cystic fibrosis, retinal disorders, psoriasis and other diseases. In the present review, we outline the history of proteases as therapeutics, provide an overview of their current clinical application, and describe several approaches to improve and expand their clinical application. Undoubtedly, our ability to harness proteolysis for disease treatment will increase with our understanding of protease biology and the molecular mechanisms responsible. New technologies for rationally engineering proteases, as well as improved delivery options, will expand greatly the potential applications of these enzymes. The recognition that proteases are, in fact, an established class of safe and efficacious drugs will stimulate investigation of additional therapeutic applications for these enzymes. Proteases therefore have a bright future as a distinct therapeutic class with diverse clinical applications.

Download full-text


Available from: Charles S Craik, Aug 11, 2015
1 Follower
  • Source
    • "Proteases control important biological processes, such as DNA replication, cell-cycle progression, cell proliferation, differentiation, and apoptosis through their ability to initiate , modulate, and terminate a variety of essential cellular functions by the processing of peptides and proteins [1]. In addition to their role in nature, proteases also serve as essential tools in industrial applications as well as biomedical research [2] [3]. There exist a number of methods to characterize and engineer proteases and protease substrates [4] [5]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Proteases are involved in many biological processes and have become important tools in biomedical research and industry. Technologies for engineering and characterization of, for example, proteolytic activity and specificity are essential in protease research. Here, we present a novel method for assessment of site-specific proteolysis. The assay utilizes plasmid-encoded reporters that, upon processing by a co-expressed protease, confer antibiotic resistance to bacteria in proportion to the cleavage efficiency. We have demonstrated that cells co-expressing cleavable reporters together with tobacco etch virus protease (TEVp) could be discriminated from cells with non-cleavable reporters by growth in selective media. Importantly, the resistance to antibiotics proved to correlate with the substrate processing efficiency. Thus, by applying competitive growth of a mock library in antibiotic-containing medium, we could show that the substrate preferred by TEVp was enriched relative to less-efficient substrates. We believe that this simple methodology will facilitate protease substrate identification, and hold great promise for directed evolution of proteases and protease recognition sequences towards improved or even new functionality.
    Biotechnology Journal 01/2014; 9(1). DOI:10.1002/biot.201300234 · 3.71 Impact Factor
  • Source
    • "There is already a large array of commercially used proteases ranging from detergent additives to effective therapeutics. The therapeutic proteases have recently been nicely reviewed [4]. The present review expands coverage to include the large variety of other commercial protease classes. "
    [Show abstract] [Hide abstract]
    ABSTRACT: This review presents a brief overview of the general categories of commercially used proteases, and critically surveys the successful strategies currently being used to improve the properties of proteases for various commercial purposes. We describe the broad application of proteases in laundry detergents, food processing, and the leather industry. The review also introduces the expanding development of proteases as a class of therapeutic agents, as well as highlighting recent progress in the field of protease engineering. The potential commercial applications of proteases are rapidly growing as recent technological advances are producing proteases with novel properties and substrate specificities.
    FEBS letters 01/2013; 587(8). DOI:10.1016/j.febslet.2012.12.019 · 3.34 Impact Factor
  • Source
    • "Amino acid substitutions in over 50 % of the 275 amino acids of subtilisins are reported (Bryan 2000; Leisola and Turunen 2007). The focus of protease engineering has shifted in the last years towards engineering of proteases for novel and precise sequence specificity cleavage for the use in analytical, biotechnological, and therapeutic applications since a single protease molecule can inactivate, due to its catalytic turnover, numerous target proteins (Craik et al. 2011; Pogson et al. 2009). For example, subtilisin BPN' was through iterative modeling, mutagenesis, and kinetic analysis cycles, tailored to preferentially cleave phosphotyrosine peptides (2,500-fold enhanced relative to the wild type; Knight et al. 2007). "
    [Show abstract] [Hide abstract]
    ABSTRACT: In proteins, a posttranslational deamidation process converts asparagine (Asn) and glutamine (Gln) residues into negatively charged aspartic (Asp) and glutamic acid (Glu), respectively. This process changes the protein net charge affecting enzyme activity, pH optimum, and stability. Understanding the principles which affect these enzyme properties would be valuable for protein engineering in general. In this work, three criteria for selecting amino acid substitutions of the deamidation type in the Bacillus gibsonii alkaline protease (BgAP) are proposed and systematically studied in their influence on pH-dependent activity and thermal resistance. Out of 113 possible surface amino acids, 18 (11 Asn and 7 Gln) residues of BgAP were selected and evaluated based on three proposed criteria: (1) The Asn or Gln residues should not be conserved, (2) should be surface exposed, and (3) neighbored by glycine. "Deamidation" in five (N97, N253, Q37, Q200, and Q256) out of eight (N97, N154, N250, N253, Q37, Q107, Q200, and Q256) amino acids meeting all criteria resulted in increased proteolytic activity. In addition, pH activity profiles of the variants N253D and Q256E and the combined variant N253DQ256E were dramatically shifted towards higher activity at lower pH (range of 8.5-10). Variant N253DQ256E showed twice the specific activity of wild-type BgAP and its thermal resistance increased by 2.4 °C at pH 8.5. These property changes suggest that mimicking surface deamidation by substituting Gln by Glu and/or Asn by Asp might be a simple and fast protein reengineering approach for modulating enzyme properties such as activity, pH optimum, and thermal resistance.
    Applied Microbiology and Biotechnology 11/2012; 97(15). DOI:10.1007/s00253-012-4560-8 · 3.81 Impact Factor
Show more