Substituted ceramidonines were synthesized by cyclization of 1‐(arylamino)anthraquinones under acidic conditions. These key intermediates were prepared either by copper‐promoted N‐arylation of 1‐aminoanthraquinone with iodoarenes or, more efficiently, by palladium‐catalyzed N‐arylation of anilines using 1‐chloroanthraquinone. The latter route was then extended to the synthesis of 1‐(arylamino)xanthones and ‐thioxanthones, which were subsequently cyclized to form (thio)chromeno[2,3,4‐kl]acridines. Taking the strategy further, stepwise N‐arylation of 1,5‐dichloroanthraquinone followed by cyclization afforded symmetrical and unsymmetrical benzo‐fused 3,9‐diazaperylenes. Electrochemical studies indicated that substituted ceramidonines are more readily reduced than anthraquinone, based on their first and second reduction potentials. Their photophysical behavior, however, proved remarkably diverse: Although some derivatives exhibited notable fluorescence, others functioned as photosensitizers, depending on the nature of their substituents. Thiochromeno‐ and chromeno[2,3,4‐kl]acridines showed only moderate fluorescence, whereas benzo[1,2,3‐kl:4,5,6‐k’l’]diacridines displayed stronger emission and promising singlet‐oxygen quantum yields. A symmetric indolo‐fused 3,9‐diazaperylene—already known for its high fluorescence—also delivered a significant two‐photon absorption cross section of 118 GM. Finally, biological assays revealed that 2‐hydroxyceramidonine and a helicoidal benzothieno analog of ceramidonine are promising inhibitors of PIM kinase, achieving IC50 values below the micromolar range.

Background:Many factors contribute to muscle impairment during chronic kidney disease, among which vitamin D deficiency has been implicated. The aim of this study was to analyze whether cholecalciferol supplementation in hemodialysis patients with low 25-hydroxyvitamin vitamin D (25OHD) concentrations could improve handgrip strength (HGS). Methods:VITADIAL (Does Correction of 25 OH-VITAmin D With Cholecalciferol Supplementation Increase Muscle Strength in HemoDIALysis Patients?) is a prospective open-labeled French multicenter study in which chronic hemodialysis patients with a 25OHD ≤50 nmol/L were randomized to receive monthly oral 100 000 IU cholecalciferol or no vitamin D treatment during 6 months to assess whether cholecalciferol supplementation in vitamin D-deficient hemodialysis patients improves muscle strength as measured with HGS. The main objective of the study was to analyze whether a 6-month period of oral cholecalciferol improved the muscle strength of hemodialysis patients with low 25OHD levels. Results:Within the 270 hemodialysis patients from 10 hemodialysis centers included in the study, 143 patients with 25OHD ≤50 nmol/L were randomized: 70 were allocated to the cholecalciferol group and 73 to the control group. The two groups were comparable at inclusion. The cholecalciferol supplemented group's 25OHD rose to 89.78 ± 33.43 nmol/L at 6 months and remained ≤50 nmol/L for most patients in the control group, but HGS did not change significantly between randomization and end of the study in both groups (0.07 ± 3.40 kg and -0.19 ± 3.35 kg, respectively, P-value .682). No difference was found in terms of either autonomy or frailty. Conclusions:This highly powered study did not find any effect of cholecalciferol supplementation in muscle strength of hemodialysis patients with low 25OHD.Trial registration: ClinicalTrials, NCT04262934. Registered 10 February 2020. Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04262934.

The physical properties of anthraquinones can be modulated by the introduction of arylamino groups next to the carbonyl function. Indeed, formation of an intramolecular hydrogen bond can enhance atom polarization and thus reduce their highest occupied molecular orbital‐lowest unoccupied molecular orbital energy gap, resulting in light absorption at longer wavelength. In this study, it successfully prepares different arylamino derivatives of anthraquinone, xanthone, and thioxanthone, compounds that are readily converted to their respective boracycles by reaction with BF3·Et2O. The effect of the BF2 moiety on the electronic properties is evaluated by electrochemical and photophysical studies. These studies respectively reveal for the complexes easier reduction and red‐shifted absorption wavelengths. In addition, promising 8% fluorescence quantum yield and 52% singlet oxygen quantum yield are recorded for one of the diboron‐anthraquinone complexes. Representative compounds are finally tested on a panel of disease‐related protein kinases.

In the past two decades, various non-apoptotic pathways of regulated cell death have been identified; a small subset of these, including necroptosis and ferroptosis, manifests the phenotypic features of necrotic death. These two regulated necroses are being extensively studied because of their putative roles in severe acute and chronic pathologies. Moreover, as these regulated necrotic pathways are coactivated in a number of common pathologies, the development of multi-target directed ligands (that is, the use of a polypharmacological strategy) is a path-breaking avenue of research. In this study, we determined that the 7-azaindole derivative, sibiriline, inhibited both RIPK1-driven necroptosis (induced by Tumor Necrosis Factor-α) and ferroptosis (triggered by various classes of ferroptosis inducers), with EC50s against each in the µM range. We next performed a combined large-scale transcriptomic study in order to determine the molecular mechanisms of action of sibiriline. We identified the stress response protein heme oxygenase-1 (HMOX1) as the main biomarker of ferroptosis inhibition by sibiriline. We hypothesized that this compound reacts as an antioxidant to block ferroptosis; indeed, we found that sibiriline inhibits lipid peroxidation by trapping phospholipid-derived peroxyl radicals as a radical-trapping antioxidant (RTA). Taken together, these results show that sibiriline is a new dual inhibitor of necroptosis and ferroptosis cell death pathways; it works by inhibition of both RIPK1 kinase and (phospho)lipid peroxidation. We also demonstrate the in vitro efficacy of sibiriline to inhibit cell death in cell-based models of Parkinson's disease and cystic fibrosis. These findings shed light on the high therapeutic potency of RIPK1 inhibitors with RTA activity.

Genetic investigations in nephrology have long been viewed as the prerogative of paediatricians or restricted to archetypal genetic nephropathies with highly penetrant variants affecting young adults. However, genetic testing has emerged as a pivotal tool in the field of adult nephrology, with the ability to revolutionize the understanding and management of adult kidney diseases. Here, we explore the multifaceted role of genomic testing (such as exome or genome sequencing) in chronic kidney disease, shedding light on current genetic findings for reframing diagnostic paradigms and tailoring treatment strategies. Genomic testing has enhanced our comprehension of kidney diseases of unknown origin by showing that 20

Based on previous results, new 1H-pyrrolo[3,2-g]isoquinolines were synthesized and evaluated for their ability to inhibit Haspin. Considering that analogues methylated at the indole nitrogen could retain their Haspin inhibitory potency, conjugates that could be suitable for a use in a PROTAC approach were prepared by N-alkylation. In addition, based on the Haspin inhibitory potency of 1H-pyrrolo[3,2-g]isoquinoline-3-carbaldehyde analogue, an additional PROTAC candidate was synthesized by carbonylation at the 3-position. Unfortunately, none of these conjugates exhibited Haspin inhibitory potency. Nevertheless, N-methylated derivative 10 bearing a pyridin-4-yl substituent at the 3-position is the best selective Haspin inhibitor identified to date in these series, with an IC50 value of 23.6 nM and a selectivity index superior to 14 compared to other protein kinases tested. Additionally, this compound showed both interesting effects on cell viability of various human cell lines and significant inhibitory properties on cellular Haspin kinase.

Our research group identified CTN1122, an imidazo[1,2-a]pyrazine derivative, as a promising antileishmanial agent targeting intramacrophage amastigotes of Leishmania major and Leishmania donovani. CTN1122 selectively inhibits Leishmania casein kinase 1 (L-CK1.2) with a favorable safety profile. Docking studies based on a homology model highlighted key pharmacophoric elements: a 4-pyridyl group at C3, crucial for hydrogen bonding with leucine 90 in the ATP-binding site, and a 4-fluorophenyl moiety at C2, fitting into a hydrophobic pocket. In order to validate these findings, 14 analogs were synthesized with targeted modifications on the imidazo[1,2-a]pyrazine core structure. Three probed the C8 position, three evaluated the impact of C2 substitution, six assessed the C3 4-pyridyl group, and two combined changes at C8 and C3. The study confirmed the critical role of C2 and C3 substituents, as their absence significantly reduced L-CK1.2 inhibition and antileishmanial activity. Additionally, the nitrogen's position within the pyridine ring at C3 proved essential: compound 23, with a meta-pyridyl group, was inactive. Notably, compound 30 exhibited the highest antileishmanial in vitro potency (IC50 = 0.20 μM for L. major; 0.16 μM for L. donovani) alongside enhanced L-CK1.2 inhibition (IC50 = 0.384 μM), with no significant mammalian cytotoxicity.

Our research group previously discovered CTN1122, an imidazo[1,2-a]pyrazine compound with promising antileishmanial activity against intramacrophage amastigotes of Leishmania major and L. donovani strains. CTN1122 effectively targets Leishmania casein kinase 1 (L-CK1.2) and exhibits a favorable safety profile. To further explore its chemical space, we developed a convergent strategy to modify the C2 position of the imidazo[1,2-a]pyrazine core using Suzuki-Miyaura coupling of the corresponding triflate intermediate. Among 15 newly synthesized analogs, seven derivatives featuring variously substituted phenyl rings at C2 demonstrated L-CK1.2 inhibition within micromolar to submicromolar ranges and antileishmanial activity in vitro with low cytotoxicity in macrophages. Compounds 7 d and 7 l were particularly potent, with IC50 values of 1.25 mu M and 0.92 mu M against L. major, and 1.44 mu M and 2.34 mu M against L. donovani, respectively. They showed IC50 L-CK1.2=0.30 mu M and 0.57 mu M with enhanced selectivity indices (SI=3.8 and 1.6) over the human CK1 & varepsilon; ortholog. Additionally, four C2 analogs and two C5 isomers exhibited notable antiparasitic effects without strongly inhibiting L-CK1.2, indicating a possible alternative mechanism of action. Compound 7 k displayed the highest general activity, with IC50 values of 0.31 mu M on L. major and 0.27 mu M on L. donovani, coupled with favorable selectivity indexes.

The carcinogenic potency categorisation approach (CPCA) has recently been introduced by health authorities. In this model, structural features from recent literature, industry proposals, and analyses performed by health authorities, provide a rapid assessment of the potential acceptable intake (AI) for a nitrosamine impurity. As with other screening regulatory values (such as the ICH M7 Threshold of Toxicological Concern), the CPCA is conservative and can be considered a de minimis risk management framework. In cases where a nitrosamine drug substance-related impurity (NDSRI) is present below the CPCA limit, the framework provides resolution from a toxicological perspective (i.e., no further toxicology studies are required). Where an NDSRI is above the CPCA limit, the framework provides for the initiation of additional activities (i.e., the CPCA is not the only possible limit). Read-across approaches are described in both the CPCA and M7 guidance and can provide a limit with more specific applicability than the general model. The use of available experimental data (in vitro or in vivo), is valuable in order to provide an even more specific limit. The CPCA provides a framework; however, data should permit changing the AI from initial structural assessment, based on increasing data, to ultimately increase precision of the AI.

Background. Alport syndromes (AS) are the second leading genetic cause of kidney failure. Whether the multiple kidney cysts (MKC) phenotype belongs to the AS spectrum remains debated. Methods. This multicenter retrospective study focused on patients genotyped with pathogenic COL4A3, COL4A4, or COL4A5 variants (classified as ACMG-AMP 4 or 5) between January 2011 and January 2023 across four French university hospitals. The study aimed to compare characteristics between two groups based on the presence or absence of MKC, defined by three or more cysts per kidney. The MKC group was compared to a control group with negative exome sequencing results for undetermined kidney disease (ES-UKD) to assess the association between MKC and AS. Results. Among the 257 AS patients included, 38 (14.8%) presented MKC without variation from hereditary cystic kidney panel. MKC showed a significant association with male gender (P = 0.004), cardiovascular risk factors, and loss of function variants (P = 0.012). Kidney failure onset appeared significantly later, by 6 years, in MKC patients (P = 0.035). Comparison with the ES-UKD (n = 990) control group showed no significant association between AS and MKC by univariate and multivariate analysis. Multivariate analysis identified patient age and male gender (P < 0.001) as factors linked to MKC. Conclusions. A 14.8% prevalence of MKC was found in our cohort of 257 patients with AS. MKC-AS patients exhibited clinical and histological characteristics akin to nephroangiosclerosis. Our comprehensive analysis, incorporating a sizable ES-UKD cohort, revealed no significant association between MKC and AS, thus questioning the inclusion of MKC within the spectrum of AS.

Protein kinase inhibitors are a key class of targeted cancer therapies, but finding compounds that show on-target efficacy with minimal toxicity remains challenging. CDK8 and CDK19 are paralogous kinases that regulate Mediator complex to promote transcriptional responses to extracellular signals and have also been reported to facilitate genome replication. Preclinical studies show that CDK8/19 inhibitors may have therapeutic benefit in several cancers, but whether effective inhibition of Mediator kinases is cytotoxic is debated. Here, we present a multi-modal approach for discovery of specific kinase inhibitors, and find that most CDK8/19 inhibitors show off-target toxicity and/or insufficient on-target engagement in cells. We describe several novel high affinity CDK8/19 inhibitors, report that loss of CDK8/19 activity is not cytotoxic but reduces cell proliferation without affecting DNA replication, and provide evidence that most inhibitors can bind in type I or type II modes. Our approach is applicable to any kinase inhibitor development program. ### Competing Interest Statement The authors have declared no competing interest. Institut National du Cancer, PLBIO15-005 La Ligue Contre le Cancer, https://ror.org/00rkrv905, EL2010.LNCC/DF, EL2013.LNCC/DF, EL2018.LNCC/DF

The cdc2-like kinases (CLKs), are a family of kinases that attracted recently the interest of scientists due to their significant biological roles, in particular in the regulation of the mRNA splicing process. Among the four isoforms of CLKs, CLK3 is the one for which the biological roles are less understood, in part because no selective inhibitor of this challenging kinase has been found to date. Based on structural analysis of the CLKs we have identified the lysine 241, present only in CLK3, as an attractive residue to design inhibitors with increased affinity towards this kinase as compared to the three other isoforms CLK1, CLK2, and CLK4. Based on this observation, we have been able to transform a molecule (DB18) previously established with a very low activity on CLK3 into a derivative VS-77 which has now a significant affinity toward CLK3 (IC50 = 0.3 μM). Thus, VS-77 appears as a new pan-inhibitor of the CLK family.

OBJECTIVE To examine the effects of insulin-adjunctive therapy with a sodium-glucose cotransporter 2 (SGLT2) inhibitor and a glucagon receptor antagonist (GRA) on glycemia, insulin use, and ketogenesis during insulinopenia in type 1 diabetes. RESEARCH DESIGN AND METHODS In a randomized, double-blind, placebo-controlled, crossover trial we assessed the effects of adjunctive SGLT2 inhibitor therapy (dapagliflozin 10 mg daily) alone and in combination with the GRA volagidemab (70 mg weekly) in 12 adults with type 1 diabetes. Continuous glucose monitoring, insulin dosing, and insulin withdrawal tests (IWT) for measurement of glucose and ketogenesis during insulinopenia were completed during insulin-only (Baseline), SGLT2 inhibitor, and combination (SGLT2 inhibitor + GRA) therapy periods. RESULTS Average glucose and percent time with glucose in range (70-180 mg/dL) improved with combination therapy versus Baseline and SGLT2 inhibitor (131 vs. 150 and 138 mg/dL [P < 0.001 and P = 0.01] and 86% vs. 70% and 78% [P < 0.001 and P = 0.03], respectively) without increased hypoglycemia. Total daily insulin use decreased with combination therapy versus Baseline and SGLT2 inhibitor (0.41 vs. 0.56 and 0.52 units/kg/day [P < 0.001 and P = 0.002]). Peak beta-hydroxybutyrate levels during IWT were lower with combination therapy than with SGLT2 inhibitor (2.0 vs. 2.4 mmol/L; P = 0.048) and similar to levels reached during the Baseline testing period (2.1 mmol/L). Participants reported enhanced treatment acceptability and satisfaction with combination therapy. CONCLUSIONS Glucagon antagonism enhances the therapeutic effects of SGLT2 inhibition in type 1 diabetes. Combination therapy improves glycemic control, reduces insulin dosing, and suggests a strategy to unlock the benefits of SGLT2 inhibitors while mitigating the risk of diabetic ketoacidosis.