A Plug-and-Play Approach for the De Novo Generation of Dually Functionalised Bispecifics

Author(s):  
Antoine Maruani ◽  
Peter A. Szijj ◽  
Calise Bahou ◽  
João C. F. Nogueira ◽  
Stephen Caddick ◽  
...  

<p>Diseases are multifactorial, with redundancies and synergies between various pathways. However, most of the antibody-based therapeutics in clinical trials and on the market interact with only one target thus limiting their efficacy. The targeting of multiple epitopes could improve the therapeutic index of treatment and counteract mechanisms of resistance. To this effect, a new class of therapeutics emerged: bispecific antibodies.</p><p>Bispecific formation using chemical methods is rare and low yielding and/or requires a large excess of one of the two proteins to avoid homodimerisation. In order for chemically prepared bispecifics to deliver their full potential, high-yielding, modular and reliable cross-linking technologies are required. Herein, we describe a novel approach not only for the rapid and high-yielding chemical generation of bispecific antibodies from native antibody fragments, but also for the site-specific dual functionalisation of the resulting bioconjugates. Based on orthogonal clickable functional groups, this strategy enables the assembly of functionalised bispecifics with controlled loading in a modular and convergent manner.</p>

2019 ◽  
Author(s):  
Antoine Maruani ◽  
Peter A. Szijj ◽  
Calise Bahou ◽  
João C. F. Nogueira ◽  
Stephen Caddick ◽  
...  

<p>Diseases are multifactorial, with redundancies and synergies between various pathways. However, most of the antibody-based therapeutics in clinical trials and on the market interact with only one target thus limiting their efficacy. The targeting of multiple epitopes could improve the therapeutic index of treatment and counteract mechanisms of resistance. To this effect, a new class of therapeutics emerged: bispecific antibodies.</p><p>Bispecific formation using chemical methods is rare and low yielding and/or requires a large excess of one of the two proteins to avoid homodimerisation. In order for chemically prepared bispecifics to deliver their full potential, high-yielding, modular and reliable cross-linking technologies are required. Herein, we describe a novel approach not only for the rapid and high-yielding chemical generation of bispecific antibodies from native antibody fragments, but also for the site-specific dual functionalisation of the resulting bioconjugates. Based on orthogonal clickable functional groups, this strategy enables the assembly of functionalised bispecifics with controlled loading in a modular and convergent manner.</p>


2021 ◽  
Vol 22 (13) ◽  
pp. 7236
Author(s):  
Endah Dwi Hartuti ◽  
Takaya Sakura ◽  
Mohammed S. O. Tagod ◽  
Eri Yoshida ◽  
Xinying Wang ◽  
...  

Plasmodium falciparum’s resistance to available antimalarial drugs highlights the need for the development of novel drugs. Pyrimidine de novo biosynthesis is a validated drug target for the prevention and treatment of malaria infection. P. falciparum dihydroorotate dehydrogenase (PfDHODH) catalyzes the oxidation of dihydroorotate to orotate and utilize ubiquinone as an electron acceptor in the fourth step of pyrimidine de novo biosynthesis. PfDHODH is targeted by the inhibitor DSM265, which binds to a hydrophobic pocket located at the N-terminus where ubiquinone binds, which is known to be structurally divergent from the mammalian orthologue. In this study, we screened 40,400 compounds from the Kyoto University chemical library against recombinant PfDHODH. These studies led to the identification of 3,4-dihydro-2H,6H-pyrimido[1,2-c][1,3]benzothiazin-6-imine and its derivatives as a new class of PfDHODH inhibitor. Moreover, the hit compounds identified in this study are selective for PfDHODH without inhibition of the human enzymes. Finally, this new scaffold of PfDHODH inhibitors showed growth inhibition activity against P. falciparum 3D7 with low toxicity to three human cell lines, providing a new starting point for antimalarial drug development.


2020 ◽  
Vol 22 (1) ◽  
pp. 279
Author(s):  
Andrzej Kuczumow ◽  
Renata Chałas ◽  
Jakub Nowak ◽  
Wojciech Smułek ◽  
Maciej Jarzębski

A series of linear profiles of the elements of the enamel in human molar teeth were made with the use of an electron microprobe and a Raman microscope. It is postulated that the enamel can be treated as the superposition of variable “overbuilt” enamel on the stable “core” enamel at the macro-, micro- and nanoscale level. The excessive values characterize the “overbuilt enamel”. All the profiles of excessive parameters along the enamel thickness from the enamel surface to the dentin enamel junction (DEJ) can be approximated very precisely with the use of exponential functions, where Ca, P, Cl and F spatial profiles are decaying while Mg, Na, K and CO32− ones are growing distributions. The “overbuilt” apatite formed on the boundary with DEJ, enriched in Na, Mg, OH and carbonates, reacts continuously with Ca, Cl and F, passing into an acid-resistant form of the “overbuilt” enamel. The apparent phases arriving in boundary regions of the “overbuilt enamel” were proposed. Microdiffraction measurements reveal relative variation of energy levels during enamel transformations. Our investigations are the milestones for a further new class of biomaterial and nanomaterial development for biomedical applications.


2018 ◽  
Vol 46 (8) ◽  
pp. 4325-4325 ◽  
Author(s):  
Thomas Germe ◽  
Judit Vörös ◽  
Frederic Jeannot ◽  
Thomas Taillier ◽  
Robert A Stavenger ◽  
...  

2017 ◽  
Vol 61 (10) ◽  
Author(s):  
Marianne Lucas-Hourani ◽  
Daniel Dauzonne ◽  
Hélène Munier-Lehmann ◽  
Samira Khiar ◽  
Sébastien Nisole ◽  
...  

ABSTRACT De novo pyrimidine biosynthesis is a key metabolic pathway involved in multiple biosynthetic processes. Here, we identified an original series of 3-(1H-indol-3-yl)-2,3-dihydro-4H-furo[3,2-c]chromen-4-one derivatives as a new class of pyrimidine biosynthesis inhibitors formed by two edge-fused polycyclic moieties. We show that identified compounds exhibit broad-spectrum antiviral activity and immunostimulatory properties, in line with recent reports linking de novo pyrimidine biosynthesis with innate defense mechanisms against viruses. Most importantly, we establish that pyrimidine deprivation can amplify the production of both type I and type III interferons by cells stimulated with retinoic acid-inducible gene 1 (RIG-I) ligands. Altogether, our results further expand the current panel of pyrimidine biosynthesis inhibitors and illustrate how the production of antiviral interferons is tightly coupled to this metabolic pathway. Functional and structural similarities between this new chemical series and dicoumarol, which was reported before to inhibit pyrimidine biosynthesis at the dihydroorotate dehydrogenase (DHODH) step, are discussed.


2018 ◽  
Author(s):  
Martin A. Fascione ◽  
Richard J. Spears ◽  
Robin L. Brabham ◽  
Darshita Budhadev ◽  
Tessa Keenan ◽  
...  

The bioconjugation of proteins with small molecules has proved an invaluable strategy for probing and perturbing dynamic biological mechanisms. The general use of chemical methods for the functionalisation of proteins remains limited however by the frequent requirement for complicated reaction partners to be present in large excess, and harsh reaction conditions which are incompatible with many protein scaffolds. Herein we describe a site-selective organocatalyst-mediated protein aldol ligation (OPAL) that affords stable carbon-carbon linked bioconjugates at neutral pH under biocompatible conditions. OPAL enables rapid chemical modification of proteins within an hour using simple aldehyde probes in minimal excess, and is utilised here in the selective affinity tagging of proteins in cell lysate. Furthermore we demonstrate that the b-hydroxy aldehyde product of the OPAL can be functionalised a second time at neutral pH in a subsequent organocatalyst-mediated oxime ligation. This tandem strategy is showcased in the ‘chemical mimicry’ of a previously inaccessible natural dual post-translationally modified protein integral to the pathogenesis of the neglected tropical disease Leishmaniasis. <br>


2021 ◽  
Author(s):  
Michael Thiel ◽  
◽  
Haifeng Wang ◽  
Dzevat Omeragic ◽  
Jean-Michel Denichou ◽  
...  

Faulting is one type of structural trap for hydrocarbon reservoirs. With more and more fields moving toward the brownfield or mature operations stage of life, the opportunity to target bypassed or attic oil in the vicinity of bounding fault(s) is becoming more and more attractive to operators. However, without an effective logging-while-drilling (LWD) tool to locate and map a fault parallel to the well trajectory, it has been challenging and potentially high risk to optimally place a well to drain oil reserves near the fault. Operators often plan these horizontal wells at a significant distance away from the mapped fault position to avoid impacts to the well construction and production of the well. Often, the interpreted fault position, based on seismic data, can have significant lateral uncertainty, and uncertainties attached to standard well survey measurements make it challenging to place the well near the fault. This often results in the wells being placed much farther from the fault than expected, which is not optimal for maximizing recovery. In other cases, due to uncertainty in the location of the fault, the wells would accidentally penetrate the side faults and cause drilling and other issues. Conventional remote boundary detection LWD tools do not assist with locating the fault position, as they only detect formation boundaries above or below the trajectory and not to the side. In this paper, the authors propose a novel approach for mapping features like a fault parallel to the well trajectory, which was previously impossible to map accurately. This new approach utilizes a new class of deep directional resistivity measurements acquired by a reservoir mapping-while-drilling tool. The deep directional resistivity measurements are input to a newly devised inversion algorithm, resulting in high-resolution reservoir mapping on the transverse plane, which is perpendicular to the well path. These new measurements have a strong sensitivity to resistivity in contrast to the sides of the wellbore, making them suitable for side fault detection. The new inversion in the transverse plane is not limited to detecting a side fault; it can also map any feature on the transverse plane to the well path, which further broadens the application of this technology. Using the deep directional resistivity data acquired from a horizontal ultra-ERD well recently drilled in the Wandoo Field offshore Western Australia, the authors tested this approach against the well results and existing control wells. Excellent mapping of the main side fault up to 30 m to the side of the well was achieved with the new approach. Furthermore, the inversion reveals other interesting features like lateral formation thickness variations and the casing of a nearby well. In addition, the methodology of utilizing this new approach for guiding geosteering parallel to side fault in real time is elaborated, and the future applications are discussed.


2018 ◽  
Vol 62 (8) ◽  
Author(s):  
Saskia du Pré ◽  
Nicola Beckmann ◽  
Mariana Cruz Almeida ◽  
Graham E. M. Sibley ◽  
Derek Law ◽  
...  

ABSTRACTF901318 (olorofim) is a novel antifungal drug that is highly active againstAspergillusspecies. Belonging to a new class of antifungals called the orotomides, F901318 targets dihydroorotate dehydrogenase (DHODH) in thede novopyrimidine biosynthesis pathway. In this study, the antifungal effects of F901318 againstAspergillus fumigatuswere investigated. Live cell imaging revealed that, at a concentration of 0.1 μg/ml, F901318 completely inhibited germination, but conidia continued to expand by isotropic growth for >120 h. When this low F901318 concentration was applied to germlings or vegetative hyphae, their elongation was completely inhibited within 10 h. Staining with the fluorescent viability dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC) showed that prolonged exposure to F901318 (>24 h) led to vegetative hyphal swelling and a decrease in hyphal viability through cell lysis. The time-dependent killing of F901318 was further confirmed by measuring the fungal biomass and growth rate in liquid culture. The ability of hyphal growth to recover in drug-free medium after 24 h of exposure to F901318 was strongly impaired compared to that of the untreated control. A longer treatment of 48 h further improved the antifungal effect of F901318. Together, the results of this study indicate that F901318 initially has a fungistatic effect onAspergillusisolates by inhibiting germination and growth, but prolonged exposure is fungicidal through hyphal swelling followed by cell lysis.


Author(s):  
Cecilie Melau ◽  
John E Nielsen ◽  
Signe Perlman ◽  
Lene Lundvall ◽  
Lea Langhoff Thuesen ◽  
...  

Abstract Context Disorders affecting adrenal steroidogenesis promote an imbalance in the normally tightly controlled secretion of mineralocorticoids, glucocorticoids, and androgens. This may lead to differences/disorders of sex development in the fetus, as seen in virilized girls with congenital adrenal hyperplasia (CAH). Despite the important endocrine function of human fetal adrenals, neither normal nor dysregulated adrenal steroidogenesis is understood in detail. Objective Due to significant differences in adrenal steroidogenesis between human and model species (except higher primates), we aimed to establish a human fetal adrenal model that enables examination of both de novo and manipulated adrenal steroidogenesis. Design and Setting Human adrenal tissue from 54 1st trimester fetuses were cultured ex vivo as intact tissue fragments for 7 or 14 days. Main Outcome Measures Model validation included examination of postculture tissue morphology, viability, apoptosis, and quantification of steroid hormones secreted to the culture media measured by liquid chromatography-tandem mass spectrometry. Results The culture approach maintained cell viability, preserved cell populations of all fetal adrenal zones, and recapitulated de novo adrenal steroidogenesis based on continued secretion of steroidogenic intermediates, glucocorticoids, and androgens. Adrenocorticotropic hormone and ketoconazole treatment of ex vivo cultured human fetal adrenal tissue resulted in the stimulation of steroidogenesis and inhibition of androgen secretion, respectively, demonstrating a treatment-specific response. Conclusions Together, these data indicate that ex vivo culture of human fetal adrenal tissue constitutes a novel approach to investigate local effects of pharmaceutical exposures or emerging therapeutic options targeting imbalanced steroidogenesis in adrenal disorders, including CAH.


Author(s):  
K. Darshana Abeyrathna ◽  
Ole-Christoffer Granmo ◽  
Xuan Zhang ◽  
Lei Jiao ◽  
Morten Goodwin

Relying simply on bitwise operators, the recently introduced Tsetlin machine (TM) has provided competitive pattern classification accuracy in several benchmarks, including text understanding. In this paper, we introduce the regression Tsetlin machine (RTM), a new class of TMs designed for continuous input and output, targeting nonlinear regression problems. In all brevity, we convert continuous input into a binary representation based on thresholding, and transform the propositional formula formed by the TM into an aggregated continuous output. Our empirical comparison of the RTM with state-of-the-art regression techniques reveals either superior or on par performance on five datasets. This article is part of the theme issue ‘Harmonizing energy-autonomous computing and intelligence’.


Sign in / Sign up

Export Citation Format

Share Document