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To confirm target engagement of hits from our high-throughput screening efforts, we ran biophysical assays on several hundreds of hits from 15 different high-throughput screening campaigns. Analyzing the biophysical assay results from these screening campaigns led us to conclude that we could be more strategic in our biophysical analysis of hits by first confirming activity in a thermal shift assay (TSA) and then confirming activity in either a surface plasmon resonance (SPR) assay or a temperature-related intensity change (TRIC) assay. To understand how this new workflow shapes the quality of the final hits, we compared TSA/SPR or TSA/TRIC confirmed and unconfirmed hits to one another using four measures of compound quality: quantitative estimate of drug-likeness (QED), Pan-Assay Interference Compounds (PAINS), promiscuity, and aqueous solubility. In general, we found that the biophysically confirmed hits performed better in the compound quality metrics than the unconfirmed hits, demonstrating that our workflow not only confirmed target engagement of the hits but also enriched for higher quality hits.

Implicit (‘unconscious’) approach–avoidance tendencies towards stimuli can be measured using the Approach Avoidance Task (AAT). We recently expanded a toolbox for analyzing the raw data of a novel, mobile version of the AAT (mAAT), that asks participants to move their phone towards their face (pull) or away (push) in response to images presented on the phone. We here tested the mAAT reaction time and the mAAT distance in a study with 71 Dutch participants that were recruited online and performed an experiment without coming to the laboratory. The participants used both the mAAT and (explicit) rating scales to respond to photographic images of food. As hypothesized, the rated wanting, rated valence and mAAT reaction time indicated a preference for palatable over unpalatable food, and for Dutch over Asian food. Additionally, as expected, arousal was rated higher for unpalatable than for palatable food, and higher for Dutch than for Asian food. The mAAT distance indicated that the unpalatable food images were moved across larger distances, regardless of the movement direction (pull or push), compared to the palatable food images; and the Dutch food images were moved across larger distances than the Asian food images. We conclude that the mAAT can be used to implicitly probe approach–avoidance motivation for complex images in the food domain. The new measure of mAAT distance may be used as an implicit measure of arousal. The ratings and the mAAT measures do not reflect the exact same information and may complement each other. Implicit measures, such as mAAT variables, are particularly valuable when response biases that can occur when using explicit ratings are expected.

To increase the chemical space around the well-known GalNAc-ligand as ASGPR-binder, a high-throughput screening campaign was performed, testing approximately 550,000 compounds. After evaluation of the potential screening hits, only one compound, which showed high similarity with guanosine nucleosides, was chosen for further profiling. Crystal structure analysis revealed the coordination of the Ca 2+ -ion within the ASGPR-binding site by the cis -diol motif of the ribose unit as well as an additional π-π-interaction of the purine heterocycle to tryptophan-243. Based on these findings, guanosine was attached via the 5'-OH group to a recently described morpholino-based nucleotide using two different linker units. The resulting morpholino-guanosine building blocks were conjugated to the 5'-end of a literature-known transthyretin targeting small interfering RNA (siRNA), leading to trivalent siRNA-guanosine conjugates, which were tested for their TTR knockdown and exhibited similar potencies as the analogous GalNAc-conjugates in vitro and in vivo.

Nicotinamide N-methyltransferase (NNMT) is a metabolic regulator that catalyzes the methylation of nicotinamide (Nam) using the co-factor S-adenosyl-L-methionine to form 1-methyl-nicotinamide (MNA). Overexpression of NNMT and the presence of the active metabolite MNA is associated with a number of diseases including metabolic disorders. We conducted a high-throughput screening campaign that led to the identification of a tricyclic core as a potential NNMT small molecule inhibitor series. Elaborate medicinal chemistry efforts were undertaken and hundreds of analogs were synthesized to understand the structure activity relationship and structure property relationship of this tricyclic series. A lead molecule, JBSNF-000028, was identified that inhibits human and mouse NNMT activity, reduces MNA levels in mouse plasma, liver and adipose tissue, and drives insulin sensitization, glucose modulation and body weight reduction in a diet-induced obese mouse model of diabetes. The co-crystal structure showed that JBSNF-000028 binds below a hairpin structural motif at the nicotinamide pocket and stacks between Tyr-204 (from Hairpin) and Leu-164 (from central domain). JBSNF-000028 was inactive against a broad panel of targets related to metabolism and safety. Interestingly, the improvement in glucose tolerance upon treatment with JBSNF-000028 was also observed in NNMT knockout mice with diet-induced obesity, pointing towards the glucose-normalizing effect that may go beyond NNMT inhibition. JBSNF-000028 can be a potential therapeutic option for metabolic disorders and developmental studies are warranted., (© 2022. The Author(s).)

The retina is considered a useful area for investigating synaptic transmission abnormalities in neuropsychiatric disorders, including as a result of using cannabis, the most widely consumed illicit substance in the developed world. The impact of regular cannabis use on retinal function has already been evaluated, using pattern and flash electroretinogram (ERG) to demonstrate a delay in ganglion and bipolar cell response. Using multifocal ERG, it was showed that the delay to be preferentially located in the central retina. ERG tests do not separately examine the impact of cannabis on the On and Off pathways. The purpose of this study is to assess On and Off pathway function using On-Off ERG. We conducted an On-Off ERG test in 42 regular cannabis users and 26 healthy controls. The protocol was compliant with the International Society for Clinical Electrophysiology of Vision (ISCEV) standards. Amplitude and peak time were measured for the a-, b- and d-waves. Results in the regular cannabis users showed a significant increase in the latencies of both the b- and the d-wave (p = 0.020, p = 0.022, respectively, Mann-Whitney U test), with no change in the wave amplitudes. A-wave peak time and amplitude were unchanged. These findings are reflective of an effect of regular cannabis use on the On and Off pathways and are consistent with previous findings which also identified increases in retinal neuron response times. We confirm here that regular cannabis use impacts the post-receptoral cones pathway at the level of bipolar cells, affecting the On and Off pathways., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Nicotinamide- N -methyltransferase (NNMT) is a cytosolic enzyme catalyzing the transfer of a methyl group from S -adenosyl-methionine (SAM) to nicotinamide (Nam). It is expressed in many tissues including the liver, adipose tissue, and skeletal muscle. Its expression in several cancer cell lines has been widely discussed in the literature, and recent work established a link between NNMT expression and metabolic diseases. Here we describe our approach to identify potent small molecule inhibitors of NNMT featuring different binding modes as elucidated by X-ray crystallographic studies.

Nonalcoholic steatohepatitis (NASH) is a progressive form of nonalcoholic fatty liver disease that can lead to irreversible liver cirrhosis and cancer. Early diagnosis of NASH is vital to detect disease before it becomes life-threatening, yet noninvasively differentiating NASH from simple steatosis is challenging. Herein, bifunctional probes have been developed that target the hepatocyte-specific asialoglycoprotein receptor (ASGPR), the expression of which decreases during NASH progression. The results show that the probes allow longitudinal, noninvasive monitoring of ASGPR levels by positron emission tomography in the newly developed rat model of NASH. The probes open new possibilities for research into early diagnosis of NASH and development of drugs to slow or reverse its progression., Competing Interests: The authors declare no conflict of interest., (© 2020 The Authors. Published by Wiley‐VCH GmbH.)

Serious adverse reactions following immunisation with adult tick-borne encephalitis (TBE) vaccines are rare, but when they occur, they most frequently involve the nervous system. We present a case of a female patient who developed a sensory and motor L4 monoradiculopathy following self-injection of an inactivated vaccine against TBE in the ipsilateral quadriceps muscle. The motor and sensory L4 dysfunction vanished after 12 months. TBE vaccine-induced radiculopathy should be considered as a mimic of spinal root compression., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Craig Kingston et al.)

In the search of a potential backup for clopidogrel, we have initiated a HTS campaign designed to identify novel reversible P2Y12 antagonists. Starting from a hit with low micromolar binding activity, we report here the main steps of the optimization process leading to the identification of the preclinical candidate SAR216471. It is a potent, highly selective, and reversible P2Y12 receptor antagonist and by far the most potent inhibitor of ADP-induced platelet aggregation among the P2Y12 antagonists described in the literature. SAR216471 displays potent in vivo antiplatelet and antithrombotic activities and has the potential to differentiate from other antiplatelet agents.

A series of novel, highly potent P2Y₁₂ antagonists as inhibitors of platelet aggregation based on a phenylpyrazole glutamic acid piperazine backbone is described. Exploration of the structural requirements of the substituents by probing the structure-activity relationship along this backbone led to the discovery of the N-acetyl-(S)-proline cyclobutyl amide moiety as a highly privileged motif. Combining the most favorable substituents led to remarkably potent P2Y₁₂ antagonists displaying not only low nanomolar binding affinity to the P2Y₁₂ receptor but also a low nanomolar inhibition of platelet aggregation in the human platelet rich plasma assay with IC₅₀ values below 50 nM. Using a homology and a three-dimensional quantitative structure-activity relationship model, a binding hypothesis elucidating the impact of several structural features was developed.

We have searched for Theta+(1540) and Xi(--)(1862) pentaquark candidates in proton-induced reactions on C, Ti, and W targets at midrapidity and square root of s = 41.6 GeV. In 2 x 10(8) inelastic events we find no evidence for narrow (sigma approximately 5 MeV) signals in the Theta+ --> pK0(S) and Xi(--) --> Xi- pi- channels; our 95% C.L. upper limits (UL) for the inclusive production cross section times branching fraction B dsigma/dy/(y approximately 0) are (4-16) mub/N for a Theta+ mass between 1521 and 1555 MeV, and 2.5 mub/N for the Xi(--). The UL of the yield ratio of Theta+/Lambda(1520) < (3-12)% is significantly lower than model predictions. Our UL of B Xi(--)/Xi(1530)0 < 4% is at variance with the results that have provided the first evidence for the Xi(--).

A 2,3,4-tri-O-pivaloylated beta-D-galactopyranosyl azide bearing a hydroxy-functionalized spacer unit at the C-6 position of the galactose was synthesized and immobilized on the solid phase by using a polymer-bound chlorosilane. The azide was reduced to the corresponding galactopyranosylamine, which served as a versatile chiral auxiliary in highly diastereoselective Ugi four-component condensation reactions at ambient temperature. Fluoride-induced cleavage from the polymeric support furnished N-glycosylated N-acylated alpha-amino acid amides. The reaction of the immobilized galactosylamine with aldehydes gave rise to the corresponding aldimines, which underwent a domino Mannich-Michael condensation reaction with Danishefsky's diene at ambient temperature to yield 2-substituted 5,6-didehydropiperidin-4-ones on the solid phase. Subsequent cleavage with tetra-n-butylammonium fluoride delivered the N-glycosylated products in high yields, purities, and diastereoselectivities. A chemoselective 1,4-hydride addition to the polymer-bound dehydropiperidinones was achieved in the presence of the bulky oxygenophilic Lewis acid methylaluminum [bis(2,6-di-tert-butyl-4-methylphenoxide)]. The conjugate addition of cyano-modified Gilman reagents to the immobilized dehydropiperidinones furnished 2,6-cis-substituted piperidine derivatives as the major diastereomers that were isolated after cleavage from the support.

To obtain mechanistic insight, detailed studies of the intriguing "spontaneous" ambient temperature ring-opening polymerization (ROP) of tin-bridged [1]ferrocenophanes Fe(eta-C(5)H(4))(2)SnR(2) 3a (R = t-Bu) and 3b (R = Mes) in solution have been performed. The investigations explored the influence of non-nucleophilic additives such as radicals and radical traps, neutral and anionic nucleophiles, Lewis acids, protic species, and other cationic electrophiles. Significantly, two novel methodologies and mechanisms for the ROP of strained [1]ferrocenophanes are proposed based on this study. First, as the addition of amine nucleophiles such as pyridine was found to strongly accelerate the polymerization rate in solution, a new nucleophilicallyassisted ROP methodology was proposed. This operates at ambient temperature in solution even in the presence of chlorosilanes but, unlike the anionic polymerization of ferrocenophanes, does not involve cyclopentadienyl anions. Second, the addition of small quantities of the electrophilic species H(+) and Bu(3)Sn(+) was found to lead to a cationic ROP process. These studies suggest that the "spontaneous" ROP of tin-bridged [1]ferrocenophanes may be a consequence of the presence of spurious, trace quantities of Lewis basic or acidic impurities. The new ROP mechanisms reported are likely to be of general significance for the ROP of other metallocenophanes (e.g., for thermal ROP in the melt) and for other metallacycles containing group 14 elements.

[structure: see text]. The concept of dipolar recoupling is introduced to 1H-1H NOESY experiments performed under HRMAS conditions. Dipole-dipole couplings are selectively recoupled during the mixing period, while MAS ensures high resolution in the spectral dimensions. Incoherent dipolar exchange is replaced by amplified coherent processes, such that time scales for polarization transfer are shortened, and dipolar double-quantum techniques become applicable. In this way, dipole-dipole couplings, as well as J-couplings, can be individually measured.

To investigate the relevance of intraoperative macroscopic evaluation of adnexal masses a prospective study was conducted from June 1st, 1993 to May 31st, 1994, which included 57 premenopausal and 60 postmenopausal women, who underwent laparotomy because of a cystic adnexal mass. The surgeons were asked to classify the tumor intraoperatively as benign or malignant and to assign to histologic groups. In addition cytology of the cyst fluid and a biopsy from the cystic wall were evaluated. Comparison of these items with the results of permanent section diagnosis revealed the tendency of the surgeons to underestimate adnexal masses depending on patients' age and the complexity of the tumor, despite of the knowledge of preoperative ultrasonographic findings. Sufficient cytolologic examination was possible in only one third of aspirates and only 21% of the examined postmenopausal malignant neoplasms have correctly been diagnosed by cytology. Evaluation of the biopsy specimens demonstrates a marked percentage of false negatives with respect to benign tumors (30% of non-functional benign neoplasms in the premenopause were assessed as functional cysts) as well as malignant neoplasms (only 72% were diagnosed correctly in the postmenopause group). In conclusion intraoperative subjective assessment, cytology and representative biopsies do not necessarily concur with the definitive histological diagnosis.

Until recently, the management of impotence was mainly concerned with the distinction of psychogenic from organic causes of erectile failure. Patients with the former were offered psychosexual counselling, while surgery, principally in the form of prosthesis implantation, was reserved for those with organic impotence. With the identification of pharmacologic regimes that can successfully induce erection, it has become necessary to distinguish between vascular and neurogenic causes of erectile failure, since it is mainly the latter patient group that can profit from an intracavernosal injection therapy with papaverine and phentolamine. Neurogenic erectile failure is further classified according to lesion localization and underlying disease.

The past 15 years have witnessed an explosion of new knowledge concerning the anatomy, neurophysiology, and pharmacology of the lower urinary tract. Knowledge of the localization of a neurologic lesion-suprapontine, suprasacral, cauda equina, or peripheral nerves-rather than the type of disease allows a more "rational approach" to the diagnosis and therapy of neurogenic voiding disorders. More specifically, advances in pharmacological understanding enable us, at least in theory, to treat voiding disorders according to their types of dysfunction: drugs to promote bladder emptying that exert their predominant effect on bladder and bladder outlet respectively, and those promoting bladder storage with the corresponding objectives. We discuss the problem that many theoretically sound drugs are, in fact, not clinically efficacious, probably due to incomplete understanding of the physiology of normal micturition and its neurogenic disorders.

We have retrospectively investigated 71 patients with ICD-9 diagnoses 293.0 or 293.1, who were treated in our clinic in 1987, with respect to the maximum severity of altered consciousness. On the basis of an available neurological grading into "clouding of consciousness", "confusional state", "delirium", "stupor", and "coma", and from knowledge of the underlying somatic disease, we have developed prognostic criteria: Disorders of heart and circulation associated with acute organic mental disorder (with altered consciousness) carry a fair prognosis for recovery of the mental disorder in approximately two-thirds of all patients affected, even in advanced age. The same statement is valid with respect to drug-induced acute organic mental disorders, whereas most cases of cerebrovascular disorders with mental disturbances proceed to chronic forms of organic mental disorder. We believe that the clinical descriptive terms referred to above allow easy grading of states of acutely/subacutely altered consciousness, thereby providing prognostic criteria for the course of the disease.