Design and evaluation of novel interferon lambda analogs with enhanced antiviral activity and improved drug attributes

Abstract

Type III interferons (IFNs) (also called IFN-λ: IFN-λ1, IFN-λ2, IFN-λ3, and IFN-λ4) are critical players in the defense against viral infection of mucosal epithelial cells, where the activity of type I IFNs is weak, and unlike type I IFNs that are associated with severe and diverse side effects, type III IFNs cause minimal side effects due to the highly restricted expression of their receptors, and thus appear to be promising agents for the treatment and prevention of respiratory and gastrointestinal viral infection. However, the antiviral potency of natural type III IFNs is weak compared to type I and, although IFN-λ3 possesses the highest bioactivity among the type III IFNs, IFN-λ1, instead of IFN-λ3, is being developed as a therapeutic drug due to the difficulty to express IFN-λ3 in the prokaryotic expression system. Here, to develop optimal IFN-λ molecules with improved drug attributes, we designed a series of IFN-λ analogs by replacing critical amino acids of IFN-λ1 with the IFN-λ3 counterparts, and vice versa. Four of the designed analogs were successfully expressed in Escherichia coli with high yield and were easily purified from inclusion bodies. Interestingly, all four analogs showed potent activity in inducing the expression of the antiviral genes MxA and OAS and two of them, analog-6 and -7, displayed an unexpected high potency that is higher than that of type I IFN (IFN-α2a) in activating the IFN-stimulated response element (ISRE)-luciferase reporter. Importantly, both analog-6 and -7 effectively inhibited replication of hepatitis C virus in Huh-7.5.1 cells, with an IC50 that is comparable to that of IFN-α2a; and consistent with the roles of IFN-λ in mucosal epithelia, both analogs potently inhibited replication of H3N2 influenza A virus in A549 cells. Together, these studies identified two IFN-λ analogs as candidates to be developed as novel antiviral biologics.

Its all about IFN‐λ4: Protective role of IFNL4 polymorphism against COVID‐19‐related pneumonia in females

“…Both are produced following the invasion of the viruses into the target cells, and despite engaging different receptor complexes, essentially initiate very similar chain reaction that generates antiviral response and mediates viral clearance 4,21 . Yet, the differences between them exist: type III IFNs (including IFN‐λ4) are produced more rapidly and more consistently, predominate in epithelial cells, act less potently and less pro‐inflammatory, and downregulate type I IFNs signaling 32,34–37 . Provided that the host is a carrier of IFNL4 rs12979860 T and/or rs368234815 ΔG allele, both HCV and SARS‐CoV‐2 infections will trigger IFN‐λ4 synthesis 33 .…”

“…4,21 Yet, the differences between them exist: type III IFNs (including IFN-λ4) are produced more rapidly and more consistently, predominate in epithelial cells, act less potently and less pro-inflammatory, and downregulate type I IFNs signaling. 32,[34][35][36][37] Provided that the host is a carrier of IFNL4 rs12979860 T and/or rs368234815 ΔG allele, both HCV and SARS-CoV-2 infections will trigger IFN-λ4 synthesis. 33 In HCV infection, where both viral clearance and viral escape mechanisms mainly involve type I IFNs, 38 the expression of fully functional IFN-λ4, due to its diminishing effect on type I IFNs activity, should benefit the virus and aggravate the prognosis.…”

Current Landscape of IFN-λ: Induction, Inhibition, and Potential Clinical Applications to Treat Respiratory Viral Infections

“…Different approaches have been designed to use IFN-λ as an efficacious therapeutic option with a more potent antiviral response than IFN type I. For example, the use of analogs of IFN-λ1 and IFN-λ3 have demonstrated high potency in activating IFN-stimulated response elements ( 80 ) as well as the use of recombinant bovine IFN-λ to treat COVID-19 ( 81 ). IFN-λ therapy of SARS-CoV-2 infection upregulated ISGs with marginal expression of the ACE2 receptor, positioning IFN-λ as a potential therapeutic agent for COVID-19 ( 82 ).…”

Clean and Folded: Optimized Production of High Quality Recombinant Human Interferon-Λ1

“…Under the conditions tested, the target protein was expressed in the form of inclusion bodies. According to the literature [5,10,16], this is a common occurrence with production of recombinant interferons in E. Coli. It should be noted that the hIFN-λ1 gene was also cloned into the pet22b+vector, at the MscI and XhoI sites, in order to create a fusion protein not only with a histidine tag at the C-terminus, but also with pelB sequence at the N-terminus.…”