Introduction Le lupus érythémateux systémique (LES) se caractérise par la production d’autoanticorps dirigés contre de multiples auto-antigènes par des cellules B auto-réactives. Les patients atteints de LES sévère réfractaire (LESsr) répondent mal aux traitements, présentent des lésions organiques progressives et ont un taux de mortalité élevé. Des séries récentes de cas ont montrées qu’un traitement par CAR-T (chimeric antigen receptor T-cell) dirigées contre le CD19 pouvait induire une rémission clinique complète chez des patients atteints de LESsr [1]. Le rapcabtagene autoleucel (YTB323) est une nouvelle thérapie CAR-T autologue dirigée contre le CD19 et de fabrication rapide par rapport aux CAR-T traditionnels, qui a été évaluée en phase 1 dans les hémopathies malignes [2]. L’objectif de cette étude était d’évaluer la sécurité, l’efficacité et la cinétique cellulaire de l’YTB323 chez des patients atteints de LESsr. Patients et méthodes Cette étude de phase 1/2 (NCT05798117) ouverte, multicentrique, mono-bras est actuellement en cours. Le traitement par YTB323 consiste en l’administration d’une dose unique de 12,5×106 cellules CAR-T après lymphodéplétion par cyclophosphamide/fludarabine. Les données de sécurité collectées comprenaient les événements indésirables, événements indésirables graves, décès, signes vitaux, électrocardiogramme et paramètres biologiques. La pharmacocinétique (PK) a été évaluée par PCR quantitative et cytométrie de flux et la pharmacodynamique (PD) en suivant l’évolution des taux de cellules B et T. L’efficacité préliminaire a été évaluée grâce aux scores d’activité de la maladie (SLEDAI), de l’évaluation globale par le médecin (PhGA) et des marqueurs rénaux et immunologiques. Résultats Entre avril 2023 et mars 2024, 11 patients atteints de LESsr ont reçu un traitement par YTB323. Nous rapportons les données préliminaires de sécurité, d’efficacité et de biomarqueurs pour ces 11 patients (date limite des données : mars 2024). Comme attendu, une cytopénie transitoire, y compris de grade 3 ou 4, a été observée chez les 11 patients, dont des anémies (n=6 ; grade 2 [n=2] ou 3 [n=4]) et des neutropénies (n=8, grade 3 [n=2] ou 4 [n=6]). Une hypogammaglobulinémie, n’ayant pas nécessité de traitement par immunoglobulines intraveineuses, est survenue chez 6 patients. Un syndrome de relargage des cytokines (CRS) a été observé chez 6 patients (grade 1 [n=3] ou 2 [n=3]) avec résolution sous tocilizumab. Un syndrome de neurotoxicité associé aux cellules immunitaires (ICANS), se manifestant par une ataxie transitoire, a été rapporté chez 1 patient (grade 2). Les complications infectieuses de grade 2 étaient une pneumonie (n=1) et une infection urinaire (n=1). Pour les 10 patients avec des données disponibles, les études de PK/PD ont montré une expansion maximale des cellules CAR-T à 12,8jours en médiane après la perfusion et une déplétion profonde des cellules B suivie d’une récupération de celles-ci par la suite. Les données préliminaires d’efficacité ont montré une réduction importante du score SLEDAI (Fig. 1A) et du PhGA, ainsi que l’amélioration des biomarqueurs de la maladie tels qu’autoanticorps (Fig. 1B), taux de complément et protéinurie. Conclusion Les données préliminaires de cet essai clinique suggèrent un profil de tolérance favorable, une expansion des cellules CAR-T, une déplétion des cellules B et une efficacité initiale de l’YTB323 dans le LESsr, justifiant la poursuite de son évaluation.

Background: Systemic lupus erythematosus (SLE) is characterised by the production of autoantibodies against multiple self-antigens by autoreactive B cells. Patients with severe refractory SLE (srSLE) exhibit failure to respond to treatments, progressive organ damage, and high mortality. Recent case reports indicated that traditionally manufactured CD19-directed chimeric antigen receptor T-cell (CAR T-cell) therapies may result in full clinical remission in srSLE.[1,2] YTB323 (rapcabtagene autoleucel) is a novel, rapidly manufactured, autologous CD19-directed CAR T-cell therapy, that demonstrated improved safety and efficacy in haematological malignancies in comparison to traditionally manufactured CAR T-cell therapies.[3] Objectives: To determine the safety, efficacy and cellular kinetics of YTB323, a CD19-directed CAR T-cell therapy, in patients with srSLE in a clinical trial setting. Methods: An open-label, single-arm, multicentre phase 1/2 study (NCT05798117) to assess the safety, efficacy and cellular kinetics of YTB323 in participants with srSLE is currently ongoing. YTB323 treatment is a single administered dose of 12.5×106 CAR-positive viable T cells following lymphodepletion with cyclophosphamide/fludarabine. Collected safety data include adverse events, severe adverse events and deaths, vital signs, electrocardiogram and laboratory assessments. Cellular kinetics data are monitored by quantitative polymerase chain reaction and flow cytometry, and pharmacodynamics assessments include levels of B cells, T cells, immunoglobulin (Ig)G, IgA, IgM, complement C3, complement C4 and autoantibodies. Preliminary efficacy is assessed using disease activity scores, Physician's Global Assessment (PhGA) and renal outcome measures. Results: Between April and December 2023, six patients with srSLE received YTB323 treatment. Safety, pharmacokinetics/pharmacodynamics (PK/PD) and preliminary efficacy data for the first three patients who received treatment (data cutoff: August 2023), and acute safety data for all six patients who received treatment (data cutoff: December 2023) are presently available. Transient lymphodepletion-related cytopenia (grade 3 or 4) was observed in all six patients as expected, including anaemia in three patients and neutropenia in five patients. Hypogammaglobulinaemia, which did not require intravenous immunoglobulin treatment, was a common adverse event. Cytokine release syndrome (CRS) was observed in four of six patients. All CRS events were grade 1 or 2 and resolved following treatment with tocilizumab. No events of immune cell-associated neurotoxicity syndrome were reported. Infectious complications included pneumonia in one patient (grade 2). For the first three patients (data cutoff: August 2023), PK/PD studies revealed peak expansion of CAR T cells approximately 13–21 days post-infusion and deep B-cell depletion, with subsequent B-cell recovery. Preliminary efficacy data for the first three patients indicated considerable reductions in SLE Disease Activity Index (SLEDAI) (Figure 1a) and PhGA (Figure 1b) accompanied by improvements in relevant disease biomarkers such as autoantibodies (Figure 1c), complement levels (Figure 2a and b), and proteinuria. Additional data from extended follow-up of all enrolled patients will be available at the next data cutoff (April 2024). Conclusion: Preliminary data from this clinical trial suggest favourable safety, CAR T-cell expansion, B-cell depletion and initial efficacy of YTB323 in srSLE, supporting its continued evaluation. Data from additional enrolled patients and continued follow-up are being generated as the study progresses. REFERENCES: [1] Mackensen A, et al. Nat Med. 2022;28:2124–32. [2] Taubmann J, et al. The EULAR Journal. 2023;93. [3] Dickinson MJ, et al. Cancer Discov. 2023;13:1982–97. Acknowledgements: The authors thank Rangariroyashe Chipika (Novartis Ireland Limited, Dublin, Ireland) and Nicola Harris (Novartis Pharmaceuticals UK Ltd, London, UK) for editorial and medical writing support, which was funded by Novartis Pharma AG, Basel, Switzerland in accordance with the Good Publication Practice (GPP 2022) guidelines (http://www.ismpp.org/gpp-2022). This study was sponsored by Novartis Pharma AG, Basel, Switzerland. Disclosure of Interests: Josefina Cortés-Hernández GSK, Astrazeneca, Otsuka, Pere Barba Jazz Pharmaceuticals, Miltenyi Biomed, Nektar, Novartis, Pierre Fabre, Allogene, Amgen, Incyte, Kite/Gilead, BMS/Celgene, Jose-Maria Alvaro-Gracia Abbvie, GSK, BMS, Novartis, Astra-Zeneca, Pfizer, UCB and Galapagos, Abbvie, GSK, BMS, Novartis, Astra-Zeneca, Pfizer, UCB and Galapagos, Mi Kwon Pfizer, Jazz, Gilead, Incyte, Julia Weinmann-Menke Astra Zeneca, Novartis, Chiesi, GSK, Boehringer-Ingelheim, Miltenyi, Vifor, Bayer, Otsuka, Takeda, Esanum, Bayer, Novartis, Boerhinger-Ingelheim, Chiesi, Astra Zeneca, Otsuka und GSK, GSK, Toray, Miltenyi, Diamed, Eva Wagner-Drouet Kite Gilead, Kite Gilead, Novartis, Takeda, MSD, Celgene, Janssen, Ozana Fischer Novartis, Novartis, Beata Kovacs Novartis, Frédérique Chaperon Novartis, David Pearson Novartis, Roche, Novartis, Tiina Kirsilä Novartis, Novartis, Novartis, Chih-Yung Sean Lee Novartis, Novartis, Clive Drakeford Novartis, Novartis, Peter Gergely Novartis, Novartis, Giulio Cavalli Novartis, SOBI, Pfizer, Novartis, Novartis, Novartis, SOBI, Pfizer, Tamas Shisha Novartis, Novartis.

The physiological role of protein kinase C (PKC) enzymes in the immune system is presented briefly. From earlier publications of others data were collected how the defects of one/two isoenzymes of PKC system suggested their involvement in the pathogenesis of human autoimmune diseases. Our observations on the defects of seven PKC isoenzymes in the peripheral blood mononuclear cells (PBMC) demonstrate that these molecular impairments are not prerequisits of the pathogenesis of systemic lupus erythematosus (SLE), mixed connective tissue disease and Sjögren's syndrome. However, these defects can modulate the disease activity and symptoms especially in SLE by several pathways. The role of PKC system in other forms of autoimmune diseases is also very small. It was of note that we detected decreased expression of PKC isoenzymes in PBMC of a European white family with an X-linked genetic background showing seasonal undulations in the lupus patient and also in her healthy mother.

L’ESSDAI et l’ESSPRI, utilisés seuls, ne permettent pas d’evaluer tous les symptômes des patients atteints de syndrome de Sjögren primitif (SSp). Le consortium NECESSITY a tout recemment développé le Sjögren's Tool for Assessing Response (STAR), un indice composite comprenant 5 domaines couvrant tous les aspects de la maladie (activité systemique, PRO, atteinte salivaire et occulaire, biologiqie). Cet outil developpé par consensus (78 experts et 20 patients) a pour but d’etre utilisé comme critère principal d’evluation de la réponse au traitement dans les futurs essais cliniques. Les objectifs de cette etude etaient d’évaluer la réponse au STAR globale, en fonction de l’activité à l’inclusion, et dans chacun des 5 domaines, ainsi que d’évaluer la valeur ajoutée du score oculaire OSS, de l’échographie des glandes salivaires (SGUS) et du facteur rhumatoïde (FR) dans la cotation du STAR. Cette étude s’appuie sur les 9 essais controlés randomisés utilisés pour le développement du STAR. L’OSS, la SGUS et le FR n’étant pas disponibles dans tous les essais, le concise STAR (cSTAR) a été calculé sans ces éléments à toutes les visites dans chaque essai. Un essai était considéré comme positif si une différence significative entre les deux bras était retrouvée pour au moins une visite. Des analyses stratifiées selon le clinESSDAI initial (< 5 vs ≥ 5) ont été réalisées chaque fois que possible. Le taux de réponse dans chaque domaine du STAR a été calculé, ainsi qu’un STAR le plus complet possible (STAR) incluant OSS, SGUS et FR lorsqu’ils étaient disponibles. La réponse SGUS a été définie comme une amélioration de 25 % du score disponible, car seul un essai a utilisé le score Hocevar. Les mêmes analyses ont été effectuées pour le Composite of Relevant Endpoints for Sjögren's Syndrome (CRESS). Le cSTAR et cCRESS ont classé de manière identique 6/9 trials. Les essais abatacept (ASAP-III), leflunomide + hydroxychloroquine (HCQ) et rituximab (RTX) (TEARS) ont été classés positifs par les 2 scores. Par contre, les essais HCQ (JOQUER) et RTX (TRACTISS) étaient classés positifs par le cCRESS uniquement et l’essai iscalimab par le cSTAR uniquement. Les analyses stratifiées selon l’activité basale ont montré une différence entre bras similaire pour chaque sous-groupe (clinESSDAI < or ≥ 5) avec cSTAR. Pour le domaine systémique, la définition du CRESS (clinESSDAI final < 5) classait comme répondeurs plus de patients avec un niveau d’activité intiale faible contrairement à la définition du STAR (diminution clinESSDAI ≥ 3 points), a conduit à une augmentation du taux de réponse globale, notamment pour les traitements à activité biologique (RTX, HCQ), même en l’absence d’amelioration du clinESSDAI. Pour les patients avec activité modérée à élevée, un taux de réponse plus important a été observé avec le STAR, mais la différence entre bras similaire que chez les patients avec faible activité initiale. L’ajout du FR aux IgG dans le domaine biologique améliorait la différence entre les bras pour 3/5 essais. L’ajout de OSS n’apportait pas d’amélioration dans les 2 essais disponibles. L’ajout du score SGUS augmentait légèrement la différence entre bras dans les 3 essais disponibles. Le cSTAR et cCRESS ont calssé de facon similaires pour la plupart des essais. Les différences observées sont liées à la définition de la réponse systémique ; CRESS classifie les patients avec activité faible comme répondeurs même en absence d’amélioration ESSDAI ou ESSPRI. L’ajout des FR et SGUS semble améliorer la réponse globale. Le STAR doit maintenant etre formellement validé dans l’essai prospectif NECESSITY avant de devenir le nouveau critère principal des essais au cours du SSp.

A foszfáttartalmú vegyületek az élő szervezetek alapvető alkotórészei. A foszfor (a periódusos rendszer 15. eleme) külső elektronhéján öt elektront tartalmaz, ezért öt kémiai (kovalens) kötést létesíthet: pl. négy oxigénatommal kapcsolódva foszfátaniont alkot. A foszfátvegyületek bőségesen előfordulnak a Földön. Vízoldékonyságuknak köszönhetően nagy koncentrációban rendelkezésre álltak az élet kialakulásának kezdetén és stabil észtereket, anhidrideket alkotva részt vettek a biológiailag fontos molekulák létrehozásában.

The design of glycogen phosphorylase (GP) inhibitors targeting the catalytic site of the enzyme is a promising strategy for a better control of hyperglycaemia in the context of type 2 diabetes. Glucopyranosylidene-spiro-heterocycles have been demonstrated as potent GP inhibitors, and more specifically spiro-oxathiazoles. A new synthetic route has now been elaborated through 1,3-dipolar cycloaddition of an aryl nitrile oxide to a glucono-thionolactone affording in one step the spiro-oxathiazole moiety. The thionolactone was obtained from the thermal rearrangement of a thiosulfinate precursor according to Fairbanks' protocols, although with a revisited outcome and also rationalised with DFT calculations. The 2-naphthyl substituted glucose-based spiro-oxathiazole 5h, identified as one of the most potent GP inhibitors (Ki = 160 nM against RMGPb) could be produced on the gram-scale from this strategy. Further evaluation in vitro using rat and human hepatocytes demonstrated that compound 5h is a anti-hyperglycaemic drug candidates performing slightly better than DAB used as a positive control. Investigation in Zucker fa/fa rat model in acute and subchronic assays further confirmed the potency of compound 5h since it lowered blood glucose levels by ∼36% at 30 mg kg-1 and ∼43% at 60 mg kg-1. The present study is one of the few in vivo investigations for glucose-based GP inhibitors and provides data in animal models for such drug candidates.

Type 2 diabetes mellitus (T2DM), one of the most common metabolic diseases, is characterized by insulin resistance and inadequate insulin secretion of β cells. Glycogen phosphorylase (GP) is the key enzyme in glycogen breakdown, and contributes to hepatic glucose production during fasting or during insulin resistance. Pharmacological GP inhibitors are potential glucose lowering agents, which may be used in T2DM therapy. A natural product isolated from the cultured broth of the fungal strain No. 138354, called 2,3-bis(4-hydroxycinnamoyloxy)glutaric acid (FR258900), was discovered a decade ago. In vivo studies showed that FR258900 significantly reduced blood glucose levels in diabetic mice. We previously showed that GP inhibitors can potently enhance the function of β cells. The purpose of this study was to assess whether an analogue of FR258900 can influence β cell function. BF142 (Meso-Dimethyl 2,3-bis[(E)-3-(4-acetoxyphenyl)prop-2-enamido]butanedioate) treatment activated the glucose-stimulated insulin secretion pathway, as indicated by enhanced glycolysis, increased mitochondrial oxidation, significantly increased ATP production, and elevated calcium influx in MIN6 cells. Furthermore, BF142 induced mTORC1-specific phosphorylation of S6K, increased levels of PDX1 and insulin protein, and increased insulin secretion. Our data suggest that BF142 can influence β cell function and can support the insulin producing ability of β cells.

3-(β-d-Glucopyranosyl)-5-substituted-1,2,4-triazoles have been revealed as an effective scaffold for the development of potent glycogen phosphorylase (GP) inhibitors but with the potency very sensitive to the nature of the alkyl/aryl 5-substituent (Kun et al., Eur. J. Med. Chem. 2014, 76, 567). For a training set of these ligands, quantum mechanics-polarized ligand docking (QM-PLD) demonstrated good potential to identify larger differences in potencies (predictive index PI = 0.82) and potent inhibitors with Ki's < 10 μM (AU-ROC = 0.86). Accordingly, in silico screening of 2335 new analogues exploiting the ZINC docking database was performed and nine predicted candidates selected for synthesis. The compounds were prepared in O-perbenzoylated forms by either ring transformation of 5-β-d-glucopyranosyl tetrazole by N-benzyl-arenecarboximidoyl chlorides, ring closure of C-(β-d-glucopyranosyl)formamidrazone with aroyl chlorides, or that of N-(β-d-glucopyranosylcarbonyl)arenethiocarboxamides by hydrazine, followed by deprotections. Kinetics experiments against rabbit muscle GPb (rmGPb) and human liver GPa (hlGPa) revealed five compounds as potent low μM inhibitors with three of these on the submicromolar range for rmGPa. X-ray crystallographic analysis sourced the potency to a combination of favorable interactions from the 1,2,4-triazole and suitable aryl substituents in the GP catalytic site. The compounds also revealed promising calculated pharmacokinetic profiles.

BackgroundAccording to international consensus, disease activity in primary Sjögren’s syndrome (pSS) patients shall be scored across 12 different domains according to the European Sjögren Syndrome Disease Activity Index (ESSDAI1). Two separate phase 2 proof-of-concept (PoC) studies using either BAFF receptor inhibitor (VAY736; ianalumab) or CD40 inhibitor (CFZ533) have recently been completed in comparable pSS cohorts with ESSDAI as the primary endpoint.ObjectivesTo evaluate contribution of individual domains to composite ESSDAI scores at baseline and after interventional treatment with either VAY736 or CFZ533, or placebo. Aggregate efficacy and safety results were presented at previous meetings. 2 3MethodsKey inclusion criteria for both studies were fulfilling revised European US consensus criteria for pSS,4 autoantibody positivity, exclusion of secondary SS and ESSDAI ≥6. Permitted background medications in both trials included stable doses of hydroxychloroquine, methotrexate and prednisone ≤10 mg/d and also azathioprine in the CFZ533 trial. The primary endpoint was change in ESSDAI at week 12. Descriptive and principal component analysis were done with the goal to identify distinct subgroups of patients based on ESSDAI involvement at baseline (BL), and the relative importance of single domain contribution to overall ESSDAI responses were explored.Results27 patients received single i.v. dose VAY736 10 mg/kg (n=12) or 3 mg/kg (n=6) or placebo, and 44 patients received multiple doses of CFZ533 10 mg/kg i.v. (n=21) or 3 mg/kg s.c. (n=8) or placebo. ESSDAI breakdown at BL showed a predominance of the articular, glandular, biological, constitutional and lymphadenopathy domains in both trials. Activity in more than 3 domains was recorded for 12/27 (44%) and 12/44 (27%) of patients in the VAY736 and CFZ533 studies, respectively. Principal component analysis identified the articular domain as the key component describing the difference between patients in their ESSDAI domains at BL. Two other domains that explained the variability between patients were the biological and glandular domains. Treatment effects in domains with low BL scores were more difficult to assess using ESSDAI. The majority of ESSDAI domains were not amenable to quantitative analysis, due to absence or low incidence at baseline.ConclusionsThe most frequently observed ESSDAI domain was articular involvement. Our results provide insights into ESSDAI domain frequency and distribution in the randomised controlled trial setting that may have implication for future trial design in pSS.Reference[1] Seror R, et al. (2016)Ann Rheum Dis75:382–389. [2] Dörner Th, et al. (2016)Arthritis Rheumatol68 Suppl 10; [3] Fisher B, et al. (2017) Arthritis Rheumatol69; Suppl 10; [4] Vitali C, et al. (2002) Ann Rheum Dis61:554–558.Disclosure of InterestT. Dörner Grant/research support from: Novartis, Charite CRO, Consultant for: Novartis, B. Fisher Consultant for: Novartis, Roche, BMS and AstraZeneca/MedImmune., X. Ren Employee of: Novartis Pharmaceuticals, P. Faerber Employee of: Novartis Pharmaceuticals, P. Gergely Shareholder of: Novartis Pharmaceuticals, Employee of: Novartis Pharmaceuticals, L. Mooney Employee of: Novartis Pharmaceuticals, Y. Li Shareholder of: Novartis Pharmaceuticals, Employee of: Novartis Pharmaceuticals, S. Oliver Shareholder of: Novartis Pharmaceuticals, Employee of: Novartis Pharmaceuticals, W. Hueber Shareholder of: Novartis Pharmaceuticals, Employee of: Novartis Pharmaceuticals, A. Wright Shareholder of: Novartis Pharmaceuticals, Employee of: Novartis Pharmaceuticals

Background and PurposeGlycogen phosphorylase (GP) is the key enzyme for glycogen degradation. GP inhibitors (GPi‐s) are glucose lowering agents that cause the accumulation of glucose in the liver as glycogen. Glycogen metabolism has implications in beta cell function. Glycogen degradation can maintain cellular glucose levels, which feeds into catabolism to maintain insulin secretion, and elevated glycogen degradation levels contribute to glucotoxicity. The purpose of this study was to assess whether influencing glycogen metabolism in beta cells by GPi‐s affects the function of these cells.Experimental ApproachThe effects of structurally different GPi‐s were investigated on MIN6 insulinoma cells and in a mouse model of diabetes.Key ResultsGPi treatment increased glycogen content and, consequently, the surface area of glycogen in MIN6 cells. Furthermore, GPi treatment induced insulin receptor β (InsRβ), Akt and p70S6K phosphorylation, as well as pancreatic and duodenal homeobox 1(PDX1) and insulin expression. In line with these findings, GPi‐s enhanced non‐stimulated and glucose‐stimulated insulin secretion in MIN6 cells. The InsRβ was shown to co‐localize with glycogen particles as confirmed by in silico screening, where components of InsR signalling were identified as glycogen‐bound proteins. GPi‐s also activated the pathway of insulin secretion, indicated by enhanced glycolysis, mitochondrial oxidation and calcium signalling. Finally, GPi‐s increased the size of islets of Langerhans and improved glucose‐induced insulin release in mice.Conclusion and ImplicationsThese data suggest that GPi‐s also target beta cells and can be repurposed as agents to preserve beta cell function or even ameliorate beta cell dysfunction in different forms of diabetes.Linked ArticlesThis article is part of a themed section on Inventing New Therapies Without Reinventing the Wheel: The Power of Drug Repurposing. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc

BackgroundPrimary Sjögren’s syndrome (pSS) is a systemic and progressive autoimmune disease characterised by lymphoid infiltration and progressive alteration of exocrine glands secretory function. Ectopic germinal center-like structures harbour plasma cells that generate autoantibodies leading to immune complex formation. Leniolisib (CDZ173) is an oral low molecular weight compound that selectively inhibits the lipid kinase PI3Kδ. In animals, leniolisib blocks PI3Kδ-dependent B cell functions, disrupts germinal centre formation and immune cell trafficking, supporting the rationale for a PI3Kδ–targeted therapy in pSSObjectivesTo evaluate the safety, tolerability, pharmacokinetics (PK) and preliminary clinical efficacy of multiple oral doses of leniolisib, after 12 weeks of treatment, in pSS patientsMethodsDouble-blind, randomised, placebo-controlled, parallel-design study recruited 30 seropositive pSS patients with moderate to severe disease activity as determined by EULAR Sjögren’s Syndrome Disease Activity Index (ESSDAI)≥6, EULAR Sjögren‘s Syndrome Patient Reported Index (ESSPRI)≥5 and stimulated whole salivary flow rate >0 mL/min. Patients were randomised in a 2:1 ratio to receive leniolisib (70 mg b.i.d) or placebo. The primary outcome was change in ESSPRI at week 12. Secondary outcomes included PK, changes in ESSDAI, the Short Form-36 (SF-36), Multidimensional Fatigue Inventory (MFI) and global Visual Analogue Scales (VAS) completed by patients and physicians. Additional assessments included lacrimal gland function and biomarkers relevant to pathway and diseaseResultsOverall safety and tolerability profile of leniolisib was acceptable, but appeared less favourable than placebo. In particular, rash occurred more frequently in the leniolisib group (11/20 patients) compared to placebo (1/10 patients). There was a slight improvement (not statistically significant) in ESSPRI scores (dryness, pain and fatigue) favouring leniolisib. Similar trends were observed in secondary endpoints (SF-36/mental and physical, MFI, VAS completed by patients and physicians). After 12 weeks of treatment, there was a slight improvement (not statistically significant) in the lacrimal gland function in the leniolisib group compared to placebo. The observed PK profile was as expected based on healthy volunteer data. Biomarker results suggest a strong and sustained target and pathway engagement, as evidenced by inhibition of phosphorylated Akt in ex-vivo stimulated B cells, significant decrease in serum CXCL13 and reduced frequency of circulating Follicular T helper-like cells. There was a trend of decreasing autoantibody levels in leniolisib-treated patientsConclusionsLeniolisib had an acceptable safety and tolerability profile, but caused rashes a known class effect of PI3K inhibitors. Target and pathway engagement were confirmed, however no clear efficacy signal for leniolisib was seen based on ESSPRI and ESSDAI in this Proof-of-Concept study at the studied dose.Disclosure of InterestT. Dörner: None declared, M. Zeher: None declared, U. Laessing: None declared, F. Chaperon: None declared, S. De Buck: None declared, A. Hasselberg: None declared, M.-A. Valentin Shareholder of: Novartis stock option, Employee of: Novartis, S. Ma: None declared, M. Cabanski: None declared, C. Kalis: None declared, C. Burkhart: None declared, P. Gergely: None declared

Aryl substituted 1-(β-d-glucosaminyl)-1,2,3-triazoles as well as C-β-d-glucosaminyl 1,2,4-triazoles and imidazoles were synthesized and tested as inhibitors against muscle and liver isoforms of glycogen phosphorylase (GP). While the N-β-d-glucosaminyl 1,2,3-triazoles showed weak or no inhibition, the C-β-d-glucosaminyl derivatives had potent activity, and the best inhibitor was the 2-(β-d-glucosaminyl)-4(5)-(2-naphthyl)-imidazole with a Ki value of 143 nM against human liver GPa. An X-ray crystallography study of the rabbit muscle GPb inhibitor complexes revealed structural features of the strong binding and offered an explanation for the differences in inhibitory potency between glucosyl and glucosaminyl derivatives and also for the differences between imidazole and 1,2,4-triazole analogues.

C-β-d-Glucopyranosyl pyrrole derivatives were prepared in the reactions of pyrrole, 2-, and 3-aryl-pyrroles with O-peracetylated β-d-glucopyranosyl trichloroacetimidate, while 2-(β-d-glucopyranosyl) indole was obtained by a cross coupling of O-perbenzylated β-d-glucopyranosyl acetylene with N-tosyl-2-iodoaniline followed by spontaneous ring closure. An improved synthesis of O-perbenzoylated 2-(β-d-glucopyranosyl) imidazoles was achieved by reacting C-glucopyranosyl formimidates with α-aminoketones. The deprotected compounds were assayed with isoforms of glycogen phosphorylase (GP) to show no activity of the pyrroles against rabbit muscle GPb. The imidazoles proved to be the best known glucose derived inhibitors of not only the muscle enzymes (both a and b) but also of the pharmacologically relevant human liver GPa (Ki = 156 and 26 nM for the 4(5)-phenyl and -(2-naphthyl) derivatives, respectively). An X-ray crystallographic study of the rmGPb-imidazole complexes revealed structural features of the strong binding, and also allowed to explain the absence of inhibition for the pyrrole derivatives.

Large scale (up to 20 g) preparation ofIandIIallowed the best inhibitors of glycogen phosphorylaseIIIto be synthesized in close to 60% overall yields fromI.

Synthetic methods were elaborated for d-glucals attached to oxadiazoles by a C–C bond. Introduction of the double bond was effected by either DBU induced elimination of PhCOOH from the O-perbenzoylated glucopyranosyl precursors or Zn/N-methylimidazole mediated reductive elimination from the 1-bromoglucopyranosyl starting compounds. Alternatively, heterocyclizations of 2-deoxy-d-arabino-hex-1-enopyranosyl cyanide were also carried out. Test compounds were obtained by Zemplén debenzoylation, however, none of them showed significant inhibition of rabbit muscle glycogen phosphorylase b.

The major role of liver glycogen is to supply glucose to the circulation maintaining the normal blood glucose level. In muscle and liver the accumulation and breakdown of glycogen are regulated by the reciprocal activities of glycogen phosphorylase and glycogen synthase. Glycogen phosphorylase catalyses the key step of glycogen degradation and its activity can be inhibited by glucose and its analogues. Obviously, any readily accessible inhibitor of glycogen phosphorylase can be used as a potential therapy of non-insulin-dependent or type 2 diabetes. Hepatic glycogen phosphorylase has been identified as a new target for drugs that control blood glucose concentration. In our experiments glucopyranosylidene-spirothiohydantoin (TH) was tested on the insulin sensitivity and blood glucose level of control and streptozotocin-treated rats. The streptozotocin-treated rats failed to gain weight and exhibited stable hyperglycemia (4.7 ± 0.5 mmol/L glucose in control vs. 7.8 ± 0.5 mmol/L) and low plasma insulin levels (9.6 ± 1.9 µIU/mL in control vs. 3.2 ± 2.2 µIU/mL). When insulin supplementation with slow-release implants (2 IU/day) was started 8 weeks after streptozotocin injection, blood glucose concentration remained suppressed, plasma insulin level dramatically increased and the insulin sensitivity restored. TH administration significantly reduced the high blood glucose concentration and restored the insulin sensitivity of STZtreated rats.

Glycogen phosphorylase (GP) is a target for the treatment of hyperglycaemia in the context of type 2 diabetes. This enzyme is responsible for the depolymerization of glycogen into glucose thereby affecting the levels of glucose in the blood stream. Twelve new d-glucopyranosylidene-spiro-isoxazolines have been prepared from O-peracylated exo-D-glucals by regio- and stereoselective 1,3-dipolar cycloaddition of nitrile oxides generated in situ by treatment of the corresponding oximes with bleach. This mild and direct procedure appeared to be applicable to a broad range of substrates. The corresponding O-unprotected spiro-isoxazolines were evaluated as glycogen phosphorylase (GP) inhibitors and exhibited IC50 values ranging from 1 to 800 μM. Selected inhibitors were further evaluated in vitro using rat and human hepatocytes and exhibited significant inhibitory properties in the primary cell culture. Interestingly, when tested with human hepatocytes, the tetra-O-acetylated spiro-isoxazoline bearing a 2-naphthyl residue showed a much lower IC50 value (2.5 μM), compared to that of the O-unprotected analog (19.95 μM). The most promising compounds were investigated in Zucker fa/fa rat model in acute and sub-chronic assays and decreased hepatic glucose production, which is known to be elevated in type 2 diabetes. This indicates that glucose-based spiro-isoxazolines can be considered as anti-hyperglycemic agents in the context of type 2 diabetes.