Current perspectives in fragment based lead discovery (FBLD)

It is over 20 years since the first fragment-based discovery projects were disclosed. The methods are now mature for most ‘conventional’ targets in drug discovery such as enzymes (kinases and proteases) but there has also been growing success on more challenging targets, such as disruption of protein–protein interactions. The main application is to identify tractable chemical startpoints that non-covalently modulate the activity of a biological molecule. In this essay, we overview current practice in the methods and discuss how they have had an impact in lead discovery – generating a large number of fragment-derived compounds that are in clinical trials and two medicines treating patients. In addition, we discuss some of the more recent applications of the methods in chemical biology – providing chemical tools to investigate biological molecules, mechanisms and systems

Fragment screening of cyclin G-associated kinase by weak affinity chromatography.

Fragment-based drug discovery (FBDD) has become a new strategy for drug discovery where lead compounds are evolved from small molecules. These fragments form low affinity interactions (dissociation constant (K(D)) = mM - μM) with protein targets, which require fragment screening methods of sufficient sensitivity. Weak affinity chromatography (WAC) is a promising new technology for fragment screening based on selective retention of fragments by a drug target. Kinases are a major pharmaceutical target, and FBDD has been successfully applied to several of these targets. In this work, we have demonstrated the potential to use WAC in combination with mass spectrometry (MS) detection for fragment screening of a kinase target-cyclin G-associated kinase (GAK). One hundred seventy fragments were selected for WAC screening by virtual screening of a commercial fragment library against the ATP-binding site of five different proteins. GAK protein was immobilized on a capillary HPLC column, and compound binding was characterized by frontal affinity chromatography. Compounds were screened in sets of 13 or 14, in combination with MS detection for enhanced throughput. Seventy-eight fragments (46 %) with K(D) < 200 μM were detected, including a few highly efficient GAK binders (K(D) of 2 μM; ligand efficiency = 0.51). Of special interest is that chiral screening by WAC may be possible, as two stereoisomeric fragments, which both contained one chiral center, demonstrated twin peaks. This ability, in combination with the robustness, sensitivity, and simplicity of WAC makes it a new method for fragment screening of considerable potential

Performance, carcass and meat traits of non-castrated cattle fed with or without monensin and/or probiotic addition to the diet

The objective of this experiment was to evaluate the performance, carcass and meat characteristics of non-castrated cattle with or without monensin (M) and/or probiotic (P) (Saccharomyces cerevisiae) to the diet. The animals were divided into individual stalls, being left 145 days in feedlot. The roughage used was corn silage + 1.2% of the weight live of concentrate in natural matter basis. There was no effect of the addition of the addictives supplied isolated or associated over feed intake, weight gain and fed conversion. Feed intake, weight gain and feed conversion averages presented quadratic behavior with the increase of the period feedlot. Although the addition of M or P to the diet resulted in the numeric increase (P>.05) of the intake (4.2%) the average of weight gain reduced (5.8 and 5.3%, respectively), resulting in smaller fed conversion (P>.05). The association of M+P already increased (P>.05) the food intake to the order of 9.5% with concomitant increase (P>.05) of the weight gain (6,4%) when compared to diet control. The animals fed with M+P presented better carcass finish (5.5mm), followed by the control (4.7mm), and smaller values, it was verified in the carcasses of the animals that received M (3.7mm) and P (3.5mm). The addition of monensin and/or probiotic for feedlot steers did not improve the performance, carcass and meat quality.O objetivo do estudo foi avaliar o desempenho e as características da carcaça e da carne de novilhos não-castrados alimentados com ou sem adição de monensina (M) e/ou probiótico (P) (Sacharomyces cerevisiae) à dieta. Os animais foram distribuídos em baias individuais, permanecendo 145 dias em confinamento. A dieta foi composta de silagem de milho e 1,2% do peso vivo de concentrado com base da matéria natural. Não houve efeito da adição dos aditivos fornecidos de forma isolada ou mesmo da associação destes sobre o consumo de alimento, o ganho de peso e a conversão alimentar. As médias de consumo, ganho de peso e conversão alimentar apresentaram comportamento quadrático com o avanço do período de confinamento. Embora a adição de M ou P à dieta tenha resultado no aumento numérico (P>0,05) do consumo (4,2%), a média de ganho de peso reduziu (5,8 e 5,3%, respectivamente), resultando em pior conversão alimentar (P>0,05). Já a associação de M+P aumentou (P>0,05) o consumo em 9,5%, com concomitante aumento (P>0,05) do ganho de peso (6,4%) em relação à dieta controle. Os animais alimentados com M+P apresentaram melhor acabamento de carcaça (5,5mm), seguidos por aqueles do grupo controle (4,7mm), sendo os valores inferiores verificados nas carcaças dos animais M (3,7mm) e P (3,5mm). A adição de monensina e/ou probiótico (Sacharomyces cerevisiae) na dieta de novilhos na fase de terminação em confinamento não proporciona melhora no desempenho e nas características da carcaça e da carne de novilhos

The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models

Avoidance of apoptosis is critical for the development and sustained growth of tumours. The pro-survival protein myeloid cell leukemia 1 (MCL1) is overexpressed in many cancers, but the development of small molecules targeting this protein that are amenable for clinical testing has been challenging. Here we describe S63845, a small molecule that specifically binds with high affinity to the BH3-binding groove of MCL1. Our mechanistic studies demonstrate that S63845 potently kills MCL1-dependent cancer cells, including multiple myeloma, leukaemia and lymphoma cells, by activating the BAX/BAK-dependent mitochondrial apoptotic pathway. In vivo, S63845 shows potent anti-tumour activity with an acceptable safety margin as a single agent in several cancers. Moreover, MCL1 inhibition, either alone or in combination with other anti-cancer drugs, proved effective against several solid cancer-derived cell lines. These results point towards MCL1 as a target for the treatment of a wide range of tumours