Improvement of Pharmacokinetic Profile of TRAIL via Trimer-Tag Enhances its Antitumor Activity in vivo

Abstract

TNF-related apoptosis-inducing ligand (TRAIL/Apo2L) has long been considered a tantalizing target for cancer therapy because it mediates activation of the extrinsic apoptosis pathway in a tumor-specific manner by binding to and trimerizing its functional receptors DR4 or DR5. Despite initial promise, both recombinant human TRAIL (native TRAIL) and dimeric DR4/DR5 agonist monoclonal antibodies (mAbs) failed in multiple human clinical trials. Here we show that in-frame fusion of human C-propeptide of α1(I) collagen (Trimer-Tag) to the C-terminus of mature human TRAIL leads to a disulfide bond-linked homotrimer which can be expressed at high levels as a secreted protein from CHO cells. The resulting TRAIL-Trimer not only retains similar bioactivity and receptor binding kinetics as native TRAIL in vitro which are 4–5 orders of magnitude superior to that of dimeric TRAIL-Fc, but also manifests more favorable pharmacokinetic and antitumor pharmacodynamic profiles in vivo than that of native TRAIL. Taken together, this work provides direct evidence for the in vivo antitumor efficacy of TRAIL being proportional to systemic drug exposure and suggests that the previous clinical failures may have been due to rapid systemic clearance of native TRAIL and poor apoptosis-inducing potency of dimeric agonist mAbs despite their long serum half-lives.

Apoptosis: A Target for Anticancer Therapy

“…Some death ligands include Fas ligand (Fas-L), TNF-related apoptosis-inducing ligand (TRAIL) and tumor necrosis factor (TNF) [ 13 ]. An adaptor protein is recruited to the death receptor [ 4 , 14 ]; adaptor proteins include Fas-associated death domain (FADD) and TNF receptor-associated death domain (TRADD) [ 13 ]. Initiator procaspases-8 and -10 bind to the adaptor protein, forming the death-inducing signaling complex (DISC) [ 4 , 14 ].…”

Progress and Prospects on Vaccine Development against SARS-CoV-2

“…For example, the University of Queensland is developing a subunit vaccine based on the "molecular clamp" technology [44]. Clover Biopharmaceuticals Inc. revealed that they are developing a vaccine candidate against SARS-CoV-2 using the "Trimer-Tag" technology [45], and the trimeric S protein subunit vaccine candidate was produced via a mammalian cell expression system. Novavax, Inc. announced that they had produced multiple nanoparticle vaccine candidates based on S protein, and now is assessing efficacy in animal models to identify an optimal vaccine candidate for human testing.…”

Ligand nanovectorization using graphene to target cellular death receptors of cancer cell

“…Their docking was then carried out in order to obtain the best match based on the total free enthalpy (or score) value. The soluble TRAIL was chosen in its cristallographic trimerized structure, because it is its most stable natural state and was used as such during docking calculations . Then, docking simulations were performed progressively by adding the receptor to the optimized TRAIL trimer, one by one.…”

“…The soluble TRAIL was chosen in its cristallographic trimerized structure, because it is its most stable natural state and was used as such during docking calculations. 35 Then, docking simulations were performed progressively by adding the receptor to the optimized TRAIL trimer, one by one. The scoring energies were then classified and the crystallographic structure was compared with the best scoring structure.…”