Targeting of Perforin Inhibitor into the Brain Parenchyma Via a Prodrug Approach Can Decrease Oxidative Stress and Neuroinflammation and Improve Cell Survival

Abstract

The cytolytic protein perforin has a crucial role in infections and tumor surveillance. Recently, it has also been associated with many brain diseases, such as neurodegenerative diseases and stroke. Therefore, inhibitors of perforin have attracted interest as novel drug candidates. We have previously reported that converting a perforin inhibitor into an L-type amino acid transporter 1 (LAT1)-utilizing prodrug can improve the compound’s brain drug delivery not only across the blood–brain barrier (BBB) but also into the brain parenchymal cells: neurons, astrocytes, and microglia. The present study evaluated whether the increased uptake into mouse primary cortical astrocytes and subsequently improvements in the cellular bioavailability of this brain-targeted perforin inhibitor prodrug could enhance its pharmacological effects, such as inhibition of production of caspase-3/-7, lipid peroxidation products and prostaglandin E2 (PGE2) in the lipopolysaccharide (LPS)-induced neuroinflammation mouse model. It was demonstrated that increased brain and cellular drug delivery could improve the ability of perforin inhibitors to elicit their pharmacological effects in the brain at nano- to picomolar levels. Furthermore, the prodrug displayed multifunctional properties since it also inhibited the activity of several key enzymes related to Alzheimer’s disease (AD), such as the β-site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1), acetylcholinesterase (AChE), and most probably also cyclooxygenases (COX) at micromolar concentrations. Therefore, this prodrug is a potential drug candidate for preventing Aβ-accumulation and ACh-depletion in addition to combatting neuroinflammation, oxidative stress, and neural apoptosis within the brain.

Potential Mechanism Underlying Exercise Upregulated Circulating Blood Exosome miR-215-5p to Prevent Necroptosis of Neuronal Cells and a Model for Early Diagnosis of Alzheimer’s Disease

“…In addition, the AD risk assessment model was closely associated with the local immune microenvironment and the pathways in this study, consistent with previous findings. Cytolytic activity-related proteins are associated with neurodegenerative diseases ( Tampio et al, 2020 ). Higher neutrophil counts are associated with AD ( Ramos-Cejudo et al, 2021 ).…”

L-type Amino Acid Transporter 1 Utilizing Ferulic Acid Derivatives Show Increased Drug Delivery in the Mouse Pancreas Along with Decreased Lipid Peroxidation and Prostaglandin Production

“…MDA was quantified by the derivatization reaction with thiobarbituric acid (TBA) (Sigma, St. Louis, MO, USA), with a previously described protocol. 30 The analysis was carried out with an Agilent 1290 Infinity II LC System (Agilent Technologies, Santa Clara, CA, USA) combined with an Agilent 1290 Infinity II Diode Array Detector. The results were calculated as μmol of formed MDA-TBA 2 per mg of tissue.…”

“…The LC–MS/MS analysis was carried out with a previously described method 30 using an Agilent 1290 Infinity LC System (Agilent Technologies, Waldbronn, Germany) and an Agilent 6495 triple quadrupole mass spectrometer with a heated electrospray ionization source (Agilent Technologies, Palo Alto, CA, USA). Data were collected using Agilent MassHunter Workstation software (version B.06.00) and processed with the Quantitative Analysis (B.07.00) software.…”

Targeting Transporters for Drug Delivery to the Brain: Can We Do Better?

“…The transport kinetic parameters, such as affinity (K m ) and transport capacity (V max ), are essential for understanding the capability of the (pro)drugs to utilize the transporter and planning pharmacokinetics studies. However, in some studies the kinetic parameters (K m and V max ) were calculated from linear or non-saturable cellular uptake data, which does not produce reliable results (119,123).…”