Discovery and SAR Study of Boronic Acid-Based Selective PDE3B Inhibitors from a Novel DNA-Encoded Library.

Human genetic evidence shows that PDE3B is associated with metabolic and dyslipidemia phenotypes. A number of PDE3 family selective inhibitors have been approved by the FDA for various indications; however, given the undesirable proarrhythmic effects in the heart, selectivity for PDE3B inhibition over closely related family members (such as PDE3A; 48% identity) is a critical consideration for development of PDE3B therapeutics. Selectivity for PDE3B over PDE3A may be achieved in a variety of ways, including properties intrinsic to the compound or tissue-selective targeting. The high (>95%) active site homology between PDE3A and B represents a massive obstacle for obtaining selectivity at the active site; however, utilization of libraries with high molecular diversity in high throughput screens may uncover selective chemical matter. Herein, we employed a DNA-encoded library screen to identify PDE3B-selective inhibitors and identified potent and selective boronic acid compounds bound at the active site.

The value of domiciliary medication reviews - a thematic analysis of pharmacist's views.

BACKGROUND: Domiciliary medication reviews are thought to enable comprehensive medication reviews centred around the needs of individuals. However, there is no clear consensus on where the value of these services lie. AIM: To determine the value of domiciliary medication reviews to service providers through semi-structured focus groups, interviews and thematic analysis. METHOD: Study participants were recruited from domiciliary medication review services provided in the United Kingdom. Semi-structured focus groups and interviews were analysed using thematic analysis. RESULTS: Six themes were identified: the scope of domiciliary medication review services, the professional role, advantages over traditional settings, disadvantages of domiciliary medication reviews for the professional, levels of engagement and outcomes. CONCLUSION: Pharmacy professionals believe that the domiciliary setting provides advantages over traditional healthcare settings when conducting medication reviews. They believe it enables a more in-depth review of an individual's medications and needs. The traditional clinical outcomes recorded by services may not be capturing the holistic impact of domiciliary medication reviews.

The value of domiciliary medication reviews: a thematic analysis of patient views.

Background Medication reviews are recognised as essential to tackling problematic polypharmacy. Domiciliary medication reviews (DMRs) have become more prevalent in recent years. They are proclaimed as being patient-centric but published literature mainly focuses on clinical outcomes. However, it is not known where the value of DMRs lies for patients who participate in them. Objective To determine the value of domiciliary medication reviews to service users. Setting Interviews took place with recipients of domiciliary medication reviews residing in the London boroughs of Islington and Haringey. Method Semi-structured interviews analysed using thematic analysis. Main outcome measure Themes and sub-themes identified from interview transcripts. Results Five themes were identified: advantages over traditional settings, attributes of the professional, adherence, levels of engagement and knowledge. Conclusion For many patients, the domiciliary setting is preferred to traditional healthcare settings. Patients appreciated the time spent with them during a DMR and felt listened to. Informal carers felt reassured that the individual medication needs of their relative had been reviewed by an expert.

The outcome of domiciliary medication reviews and their impact: a systematic review.

OBJECTIVES: Medication reviews in the domiciliary setting are becoming more prevalent internationally. Understanding the benefits of these reviews is essential to ensuring quality healthcare services. To date there has not been a systematic evaluation of the outcomes of these services and their impact on patients. A systematic review of the literature was undertaken with a view to understanding the impact of medication reviews in this setting. Controlled and uncontrolled studies were included. Outcomes were categorised according to the ECHO model. A narrative synthesis was developed. KEY FINDINGS: Nineteen out of 31 papers included demonstrated an improvement in outcome. Clinical outcomes were the most commonly measured and humanistic outcomes the least commonly measured. Domiciliary medication reviews (DMRs) services are presented as providing benefit. However, it is difficult to quantify the impact of services from the published outcomes. SUMMARY: Future work should focus on demonstrating the meaningful changes to patients that DMRs have enabled.

Modular Protein Ligation: A New Paradigm as a Reagent Platform for Pre-Clinical Drug Discovery.

Significant resource is spent by drug discovery project teams to generate numerous, yet unique target constructs for the multiple platforms used to drive drug discovery programs including: functional assays, biophysical studies, structural biology, and biochemical high throughput screening campaigns. To improve this process, we developed Modular Protein Ligation (MPL), a combinatorial reagent platform utilizing Expressed Protein Ligation to site-specifically label proteins at the C-terminus with a variety of cysteine-lysine dipeptide conjugates. Historically, such proteins have been chemically labeled non-specifically through surface amino acids. To demonstrate the feasibility of this approach, we first applied MPL to proteins of varying size in different target classes using different recombinant protein expression systems, which were then evaluated in several different downstream assays. A key advantage to the implementation of this paradigm is that one construct can generate multiple final products, significantly streamlining the reagent generation for multiple early drug discovery project teams.

A Scalable Platform for Producing Recombinant Nucleosomes with Codified Histone Methyltransferase Substrate Preferences.

Wolf-Hirschhorn Syndrome Candidate 1 (WHSC1; also known as NSD2) is a SET domain-containing histone lysine methyltransferase. A chromosomal translocation occurs in 15-20% of multiple myeloma patients and is associated with increased production of WHSC1 and poor clinical prognosis. To define the substrate requirements of NSD2, we established a platform for the large-scale production of recombinant polynucleosomes, based on authentic human histone proteins, expressed in E. coli, and complexed with linearized DNA. A brief survey of methyltransferases whose substrate requirements are recorded in the literature yielded expected results, lending credence to the fitness of our approach. This platform was readily 'codified' with respect to both position and extent of methylation at histone 3 lysines 18 and 36 and led to the conclusion that the most readily discernible activity of NSD2 in contact with a nucleosome substrate is dimethylation of histone 3 lysine 36. We further explored reaction mechanism, and conclude a processive, rather than distributive mechanism best describes the interaction of NSD2 with intact nucleosome substrates. The methods developed feature scale and flexibility and are suited to thorough pharmaceutical-scale drug discovery campaigns.

High throughput screening identifies ATP-competitive inhibitors of the NLRP1 inflammasome.

Nod-like receptors (NLRs) are cytoplasmic pattern recognition receptors that are promising targets for the development of anti-inflammatory therapeutics. Drug discovery efforts targeting NLRs have been hampered by their inherent tendency to form aggregates making protein generation and the development of screening assays very challenging. Herein we report the results of an HTS screen of NLR family member NLRP1 (NLR family, pyrin domain-containing 1) which was achieved through the large scale generation of recombinant GST-His-Thrombin-NLRP1 protein. The screen led to the identification of a diverse set of ATP competitive inhibitors with micromolar potencies. Activity of these hits was confirmed in a FP binding assay, and two homology models were employed to predict the possible binding mode of the leading series and facilitate further lead-optimization. These results highlight a promising strategy for the identification of inhibitors of NLR family members which are rapidly emerging as key drivers of inflammation in human disease.

Development of a high-throughput screen to detect inhibitors of TRPS1 sumoylation.

Small ubiquitin-like modifier (SUMO) belongs to the family of ubiquitin-like proteins (Ubls) that can be reversibly conjugated to target-specific lysines on substrate proteins. Although covalently sumoylated products are readily detectible in gel-based assays, there has been little progress toward the development of robust quantitative sumoylation assay formats for the evaluation of large compound libraries. In an effort to identify inhibitors of ubiquitin carrier protein 9 (Ubc9)-dependent sumoylation, a high-throughput fluorescence polarization assay was developed, which allows detection of Lys-1201 sumoylation, corresponding to the major site of functional sumoylation within the transcriptional repressor trichorhino-phalangeal syndrome type I protein (TRPS1). A minimal hexapeptide substrate peptide, TMR-VVK1201TEK, was used in this assay format to afford high-throughput screening of the GlaxoSmithKline diversity compound collection. A total of 728 hits were confirmed but no specific noncovalent inhibitors of Ubc9 dependent trans-sumoylation were found. However, several diaminopyrimidine compounds were identified as inhibitors in the assay with IC50 values of 12.5 µM. These were further characterized to be competent substrates which were subject to sumoylation by SUMO-Ubc9 and which were competitive with the sumoylation of the TRPS1 peptide substrates.