Therapeutic potential of polypeptide-based conjugates: Rational design and analytical tools that can boost clinical translation

SummaryHirulog™ (BG8967) is a direct thrombin inhibitor built by rational design using the protein hirudin as a model (Maraganore et al. [1990]; Biochemistry 29: 7095–101). In order to evaluate the therapeutic potential for hirulog in the management of thrombotic disease, the tolerability and anticoagulant activity of the agent were examined in a study of human volunteers.In a randomized, placebo-controlled study (n = 54), the intravenous infusion of hirulog over 15 min showed a rapid, dose-dependent prolongation of activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). There was a corresponding dose-dependent increase in plasma hirulog levels. The peptide was rapidly cleared with a half-life of 36 min and a total body clearance rate for the peptide of 0.43 1 kg−1 h−1. Similar activity was observed following subcutaneous injection but with sustained pharmacodynamic and pharmacokinetic behavior. There was a significant correlation between pharmacokinetic and pharmacodynamic variables for both intravenous (r = 0.8, p <0.001) and subcutaneous administration (r = 0.7, p = 0.002).To evaluate the possible interactions of aspirin on the tolerability and anticoagulant activity of intravenous hirulog, a cross-over design was employed in eight subjects. Aspirin administration did not modify the peptide’s activity. At the administered dose of 0.6 mg kg−1 h−1 for 2 h, hirulog infusion prolonged APTT from 230 to 260% baseline. The infusion of hirulog in subjects who had received aspirin was not associated with any significant changes in the template bleeding time.The final phase of the study examined the activity and tolerability of hirulog in ten subjects during prolonged intravenous infusions for up to 24 h. The peptide (0.3 mg kg−1 h−1) exhibited sustained anticoagulant activity with no evidence for a cumulative effect. During hirulog infusion, APTT was prolonged from 210 to 250% baseline.In all phases of the study, hirulog administration was generally well-tolerated.Our observations show that hirulog is an active antithrombin agent with excellent tolerability in humans. As a direct thrombin inhibitor, hirulog provides a novel approach for the management of thrombotic disease.

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Towards Clinical Translation: Optimized Fabrication of Controlled Nanostructures on Implant-Relevant Curved Zirconium Surfaces

Nanomaterials ◽

10.3390/nano11040868 ◽

2021 ◽

Vol 11 (4) ◽

pp. 868

Author(s):

Divya Chopra ◽

Karan Gulati ◽

Sašo Ivanovski

Keyword(s):

Therapeutic Potential ◽

Local Therapy ◽

Clinical Translation ◽

Curved Surfaces ◽

Nanoporous Layer

Anodization enables fabrication of controlled nanotopographies on Ti implants to offer tailorable bioactivity and local therapy. However, anodization of Zr implants to fabricate ZrO2 nanostructures remains underexplored and are limited to the modification of easy-to-manage flat Zr foils, which do not represent the shape of clinically used implants. In this pioneering study, we report extensive optimization of various nanostructures on implant-relevant micro-rough Zr curved surfaces, bringing this technology closer to clinical translation. Further, we explore the use of sonication to remove the top nanoporous layer to reveal the underlying nanotubes. Nano-engineered Zr surfaces can be applied towards enhancing the bioactivity and therapeutic potential of conventional Zr-based implants.

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G4Killer web application: a tool to design G-quadruplex mutations

Bioinformatics ◽

10.1093/bioinformatics/btaa057 ◽

2020 ◽

Vol 36 (10) ◽

pp. 3246-3247

Author(s):

Vaclav Brazda ◽

Jan Kolomaznik ◽

Jean-Louis Mergny ◽

Jiri Stastny

Keyword(s):

Dna Sequences ◽

Web Application ◽

Rational Design ◽

Therapeutic Potential ◽

Supplementary Information ◽

Web Tool ◽

Web Based ◽

Dna Structures ◽

G Quadruplex ◽

User Friendly

Abstract Motivation G-quadruplexes (G4) are important regulatory non-B DNA structures with therapeutic potential. A tool for rational design of mutations leading to decreased propensity for G4 formation should be useful in studying G4 functions. Although tools exist for G4 prediction, no easily accessible tool for the rational design of G4 mutations has been available. Results We developed a web-based tool termed G4Killer that is based on the G4Hunter algorithm. This new tool is a platform-independent and user-friendly application to design mutations crippling G4 propensity in a parsimonious way (i.e., keeping the primary sequence as close as possible to the original one). The tool is integrated into our DNA analyzer server and allows for generating mutated DNA sequences having the desired lowered G4Hunter score with minimal mutation steps. Availability and implementation The G4Killer web tool can be accessed at: http://bioinformatics.ibp.cz. Supplementary information Supplementary data are available at Bioinformatics online.

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Therapeutic potential of TMS-induced plasticity in the prefrontal cortex

10.1093/oxfordhb/9780198568926.013.0038 ◽

2012 ◽

Author(s):

Stanislav R. Vorel ◽

Sarah H. Lisanby

Keyword(s):

Synaptic Plasticity ◽

Rational Design ◽

Therapeutic Potential ◽

Obsessive Compulsive ◽

Neural Structure ◽

Human Motor Cortex ◽

Compulsive Disorder ◽

Human Motor ◽

And Function ◽

Pharmacological Manipulations

This article discusses synaptic plasticity as a potential mechanism of enduring changes in function observed after relatively brief periods of repetitive (r)TMS. Plasticity is a use dependent enduring change in neural structure and function. The characteristics of plasticity are described in this article. Taking into account, the interactions between rTMS and pharmacological manipulations, this article explores how principles of synaptic plasticity may be exploited in the rational design of future rTMS paradigms in psychiatric disorders like major depressive disorder, obsessive-compulsive disorder, substance use disorders, schizophrenia etc. TMS is under active study in the treatment of a range of psychiatric and neurological disorders. Furthermore, this article discusses the implications for the interpretation of existing TMS literature and design of future interventions. TMS experiments of plasticity in the human motor cortex have been limited by the intensity and frequency of TMS protocols.

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Rational Design of New Cationic Mn Porphyrins and Evaluation of Their Therapeutic Potential

Free Radical Biology and Medicine ◽

10.1016/j.freeradbiomed.2014.10.326 ◽

2014 ◽

Vol 76 ◽

pp. S94

Author(s):

Artak Tovmasyan ◽

Sebastian Carballal ◽

Robert Ghazaryan ◽

Lida Melikyan ◽

Tin Weitner ◽

...

Keyword(s):

Rational Design ◽

Therapeutic Potential ◽

Mn Porphyrins

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A Novel, Nonpeptidic, Orally Active Bivalent Inhibitor of Human β-Tryptase

Pharmacology ◽

10.1159/000492078 ◽

2018 ◽

Vol 102 (5-6) ◽

pp. 233-243 ◽

Cited By ~ 2

Author(s):

Sarah F. Giardina ◽

Douglas S. Werner ◽

Maneesh Pingle ◽

Donald E. Bergstrom ◽

Lee D. Arnold ◽

...

Keyword(s):

Active Sites ◽

Rational Design ◽

Therapeutic Potential ◽

X Ray Crystallography ◽

Inflammatory Stimuli ◽

Invaluable Tool ◽

Xenograft Models ◽

Orally Bioavailable

β-Tryptase is released from mast cells upon degranulation in response to allergic and inflammatory stimuli. Human tryptase is a homotetrameric serine protease with 4 identical active sites directed toward a central pore. These active sites present an optimized scenario for the rational design of bivalent inhibitors, which bridge 2 adjacent active sites. Using (3-[1-acylpiperidin-4-yl]phenyl)methanamine as the pharmacophoric core and a disiloxane linker to span 2 active sites we have successfully produced a novel bivalent tryptase inhibitor, compound 1a, with a comparable profile to previously described inhibitors. Pharmacological properties of compound 1a were studied in a range of in vitro enzymic and cellular screening assays, and in vivo xenograft models. This non-peptide inhibitor of tryptase demonstrated superior activity (IC50 at 100 pmol/L tryptase = 1.82 nmol/L) compared to monomeric modes of inhibition. X-ray crystallography validated the dimeric mechanism of inhibition, and 1a demonstrated good oral bioavailability and efficacy in HMC-1 xenograft models. Furthermore, compound 1a demonstrated extremely slow off rates and high selectivity against-related proteases. This highly potent, orally bioavailable and selective inhibitor of human tryptase will be an invaluable tool in future studies to explore the therapeutic potential of attenuating the activity of this elusive target.

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Linking sequence patterns and functionality of alpha-helical antimicrobial peptides

Bioinformatics ◽

10.1093/bioinformatics/bty1048 ◽

2018 ◽

Vol 35 (16) ◽

pp. 2713-2717 ◽

Cited By ~ 2

Author(s):

Igor E Eliseev ◽

Ivan N Terterov ◽

Anna N Yudenko ◽

Olga V Shamova

Keyword(s):

Antimicrobial Peptides ◽

Rational Design ◽

Therapeutic Potential ◽

Deep Understanding ◽

Supplementary Information ◽

Helical Conformation ◽

Efficient Design ◽

Sequence Patterns ◽

Functional Features ◽

Natural Peptides

Abstract Motivation The rational design of antimicrobial peptides (AMPs) with increased therapeutic potential requires deep understanding of the determinants of their activities. Inspired by the computational linguistic approach, we hypothesized that sequence patterns may encode the functional features of AMPs. Results We found that α-helical and β-sheet peptides have non-intersecting pattern sets and therefore constructed new sequence templates using only helical patterns. Designed peptides adopted an α-helical conformation upon binding to lipids, confirming that the method captures structural and biophysical properties. In the antimicrobial assay, 5 of 7 designed peptides exhibited activity against Gram(+) and Gram(–) bacteria, with most potent candidate comparable to best natural peptides. We thus conclude that sequence patterns comprise the structural and functional features of α-helical AMPs and guide their efficient design. Supplementary information Supplementary data are available at Bioinformatics online.

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Critical Evaluation of the Interaction of Reactive Oxygen and Nitrogen Species with Blood to Inform the Clinical Translation of Nonthermal Plasma Therapy

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/9750206 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Abraham Lin ◽

Eline Biscop ◽

Colum Breen ◽

Stephen J. Butler ◽

Evelien Smits ◽

...

Keyword(s):

Whole Blood ◽

Therapeutic Potential ◽

Ambient Pressure ◽

Critical Evaluation ◽

Reactive Oxygen ◽

Clinical Translation ◽

Therapeutic Effects ◽

Nitrogen Species ◽

Preclinical Research ◽

Over Time

Non-thermal plasma (NTP), an ionized gas generated at ambient pressure and temperature, has been an emerging technology for medical applications. Through controlled delivery of reactive oxygen and nitrogen species (ROS/RNS), NTP can elicit hormetic cellular responses, thus stimulating broad therapeutic effects. To enable clinical translation of the promising preclinical research into NTP therapy, a deeper understanding of NTP interactions with clinical substrates is profoundly needed. Since NTP-generated ROS/RNS will inevitably interact with blood in several clinical contexts, understanding their stability in this system is crucial. In this study, two medically relevant NTP delivery modalities were used to assess the stability of NTP-generated ROS/RNS in three aqueous solutions with increasing organic complexities: phosphate-buffered saline (PBS), blood plasma (BP), and processed whole blood. NTP-generated RNS collectively (NO2−, ONOO−), H2O2, and ONOO− exclusively were analyzed over time. We demonstrated that NTP-generated RNS and H2O2 were stable in PBS but scavenged by different components of the blood. While RNS remained stable in BP after initial scavenging effects, it was completely reduced in processed whole blood. On the other hand, H2O2 was completely scavenged in both liquids over time. Our previously developed luminescent probe europium(III) was used for precision measurement of ONOO− concentration. NTP-generated ONOO− was detected in all three liquids for up to at least 30 seconds, thus highlighting its therapeutic potential. Based on our results, we discussed the necessary considerations to choose the most optimal NTP modality for delivery of ROS/RNS to and via blood in the clinical context.

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