Safety and biodistribution of Nanoligomers^™ targeting SARS-CoV-2 genome for treatment of COVID-19

Abstract

As the world braces to enter its third year in the coronavirus disease 2019 (COVID-19) pandemic, the need for accessible and effective antiviral therapeutics continues to be felt globally. The recent surge of Omicron variant cases has demonstrated that vaccination and prevention alone cannot quell the spread of highly transmissible variants. A safe and nontoxic therapeutic with an adaptable design to respond to the emergence of new variants is critical for transitioning to treatment of COVID-19 as an endemic disease. Here, we present a novel compound, called SBCoV202, that specifically and tightly binds the translation initiation site of RNA-dependent RNA polymerase within the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome, inhibiting viral replication. SBCoV202 is a Nanoligomer,™ a molecule that includes peptide nucleic acid sequences capable of binding viral RNA with single-base-pair specificity to accurately target the viral genome. The compound has been shown to be safe and nontoxic in mice, with favorable biodistribution, and has shown efficacy against SARS-CoV-2 in vitro. Safety and biodistribution were assessed after three separate administration methods, namely intranasal, intravenous, and intraperitoneal. Safety studies showed the Nanoligomer caused no outward distress, immunogenicity, or organ tissue damage, measured through observation of behavior and body weight, serum levels of cytokines, and histopathology of fixed tissue, respectively. SBCoV202 was evenly biodistributed throughout the body, with most tissues measuring Nanoligomer concentrations well above the compound KD of 3.37 nM. In addition to favorable availability to organs such as the lungs, lymph nodes, liver, and spleen, the compound circulated through the blood and was rapidly cleared through the renal and urinary systems. The favorable biodistribution and lack of immunogenicity and toxicity set Nanoligomers apart from other antisense therapies, while the adaptability of the nucleic acid sequence of Nanoligomers provides a defense against future emergence of drug resistance, making these molecules an attractive potential treatment for COVID-19.

Targeting Neuroinflammation by Pharmacologic Downregulation of Inflammatory Pathways Is Neuroprotective in Protein Misfolding Disorders

“…Penetration of SB_NI_112 across the BBB was confirmed with ICP−MS and detected in the hippocampus (76.1 nM ± 44.3), cortex (49.9 nM ± 11.4), cerebellum, (96.9 nM ± 45.0), and brainstem (96.6 nM ± 32.4) (Figure 2A), which is significantly higher than the dissociation constant of SB_NI_112. 50 Downregulation of NF-κB and NLRP3 with SB_NI_112 leads to dampening of chronic inflammation, while preserving key immune pathways needed to respond to infectious pathogens (63). Despite downregulation of NF-κB and NLRP3, interferon gamma (IFN-γ) expression was significantly higher in prion-infected mice treated with SB_NI_112 (p = 0.048 (two-way t-test, Excel) or p = 0.0468 (one-way ANOVA, Graphpad Prism)) and not significantly different from control NBH mice (Figure 2B).…”

“…Nanoligomers were designed and synthesized (Sachi Bioworks Inc.) according to published methods. 42,43,50,75,76 The nanoligomer is composed of a nanobiohybrid molecule based on antisense peptide nucleic acid (PNA) 42,43,75,77,78 conjugated to nanoparticle 1,2,5 for improved delivery. The PNA moiety was chosen to be 17 bases long (to optimize solubility and specificity) antisense to the start codon regions of nfkb1 (N terminus-C terminus, sequence: AGTGG-TACCGTCTGCTA) and nlrp3 (N terminus-C terminus, sequence: CTTCTACTGCTCACAGG) within the mouse genome (GCF_000001635.27).…”

Inflammasome-Inhibiting Nanoligomers Are Neuroprotective against Space-Induced Pathology in Healthy and Diseased Three-Dimensional Human Motor and Prefrontal Cortex Brain Organoids

“…Additionally, these methods can also suffer from transport challenges, especially delivery to the brain across the blood-brain barrier, making molecular optimization even more challenging and reducing throughput. By contrast, the Nanoligomer therapeutic platform generates safe, targeted, 34 and highly specific (K D 3.37 nM 35,36 ) RNA-targeting molecules for facile delivery to the brain and other targeted organs and offers both up-and downregulation of desired protein through translational and transcriptional regulation. 37−42 The platform also utilizes an effective bioinformatics approach to identify top molecules for a given target, now allowing high-throughput screening and ranking of different potential targets for a given disease condition, leading to best-in-class and first-in-class therapeutics.…”

“…Nanoligomers utilize peptide nucleic acids (PNAs) as the sequence-specific binding component that demonstrate strong hybridization and specificity to their RNA (or DNA) sequence targets, 43 and exhibit no known enzymatic cleavage, leading to increased stability in human blood serum and mammalian cellular extracts. 44 Given the low K D , 35,36,43 high binding specificity, minimal off-targeting, 35,36 lack of any immunogenic response or accumulation in first pass organs, 34 absence of any observable histological damage to organs even for long (>15− 20 weeks) treatments, [34][35][36]40 and facile delivery to the brain to counter neuroinflammation, 34,37−42 we have demonstrated that Nanoligomers are a safe and targeted approach to screen for therapeutic targets in organoids and rodents for multiple neurodegenerative diseases aboard the International Space Station (ISS). Therefore, not only is the Nanoligomer-based screening well-differentiated from other traditional screening methods on Earth, but the combination of accelerated neuropathology in space and high-throughput screening by the Nanoliogmer platform can also lead to breakthrough therapeutics for different debilitating disease conditions ■ RESULTS AND DISCUSSION High-Throughput In Vitro Screening in Donor-Derived Primary Human Astrocytes.…”