7 research outputs found
Tideglusib, a Non-ATP Competitive Inhibitor of GSK-3ÎČ as a Drug Candidate for the Treatment of Amyotrophic Lateral Sclerosis
Amyotrophic Lateral Sclerosis (ALS) is the most common degenerative motor neuron disease in adults. About 97% of ALS patients present TDP-43 aggregates with post-translational modifications, such as hyperphosphorylation, in the cytoplasm of affected cells. GSK-3ÎČ is one of the protein kinases involved in TDP-43 phosphorylation. Up-regulation of its expression and activity is reported on spinal cord and cortex tissues of ALS patients. Here, we propose the repurposing of Tideglusib, an in-house non-ATP competitive GSK-3ÎČ inhibitor that is currently in clinical trials for autism and myotonic dystrophy, as a promising therapeutic strategy for ALS. With this aim we have evaluated the efficacy of Tideglusib in different experimental ALS models both in vitro and in vivo. Moreover, we observed that GSK-3ÎČ activity is increased in lymphoblasts from sporadic ALS patients, with a simultaneous increase in TDP-43 phosphorylation and cytosolic TDP-43 accumulation. Treatment with Tideglusib decreased not only phospho-TDP-43 levels but also recovered its nuclear localization in ALS lymphoblasts and in a human TDP-43 neuroblastoma model. Additionally, we found that chronic oral treatment with Tideglusib is able to reduce the increased TDP-43 phosphorylation in the spinal cord of Prp-hTDP-43A315T mouse model. Therefore, we consider Tideglusib as a promising drug candidate for ALS, being proposed to start a clinical trial phase II by the end of the year
Preclinical Investigation in Neuroprotective Effects of the GPR55 Ligand VCE-006.1 in Experimental Models of Parkinsonâs Disease and Amyotrophic Lateral Sclerosis
Cannabinoids act as pleiotropic compounds exerting, among others, a broad-spectrum of
neuroprotective effects. These effects have been investigated in the last years in different preclinical
models of neurodegeneration, with the cannabinoid type-1 (CB1) and type-2 (CB2) receptors concen-
trating an important part of this research. However, the issue has also been extended to additional
targets that are also active for cannabinoids, such as the orphan G-protein receptor 55 (GPR55). In
the present study, we investigated the neuroprotective potential of VCE-006.1, a chromenopyrazole
derivative with biased orthosteric and positive allosteric modulator activity at GPR55, in murine
models of two neurodegenerative diseases. First, we proved that VCE-006.1 alone could induce
ERK1/2 activation and calcium mobilization, as well as increase cAMP response but only in the
presence of lysophosphatidyl inositol. Next, we investigated this compound administered chronically
in two neurotoxin-based models of Parkinsonâs disease (PD), as well as in some cell-based models.
VCE-006.1 was active in reversing the motor defects caused by 6-hydroxydopamine (6-OHDA) in
the pole and the cylinder rearing tests, as well as the losses in tyrosine hydroxylase-containing
neurons and the elevated glial reactivity detected in the substantia nigra. Similar cytoprotective
effects were found in vitro in SH-SY5Y cells exposed to 6-OHDA. We also investigated VCE-006.1
in LPS-lesioned mice with similar beneficial effects, except against glial reactivity and associated
inflammatory events, which remained unaltered, a fact confirmed in BV2 cells treated with LPS and
VCE-006.1. We also analyzed GPR55 in these in vivo models with no changes in its gene expression,
although GPR55 was down-regulated in BV2 cells treated with LPS, which may explain the lack
of efficacy of VCE-006.1 in such an assay. Furthermore, we investigated VCE-006.1 in two genetic
models of amyotrophic lateral sclerosis (ALS), mutant SOD1, or TDP-43 transgenic mice. Neither the
neurological decline nor the deteriorated rotarod performance were prevented with this compound,
and the same happened with the elevated microglial and astroglial reactivities, albeit modest spinal
motor neuron preservation was achieved in both models. We also analyzed GPR55 in these in vivo
models and found no changes in both TDP-43 transgenic and mSOD1 mice. Therefore, our findings
support the view that targeting the GPR55 may afford neuroprotection in experimental PD, but not
n ALS, thus stressing the specificities for the development of cannabinoid-based therapies in the
different neurodegenerative disorders.This work has been supported by grants from CIBERNED (CB06/05/0089), MICIU (RTI-
2018-098885-B-100), ELA-Madrid-CM (B2017/BMD-3813), and Emerald Health Biotechnology-Spain.
These agencies had no further role in study design, the collection, analysis, and interpretation of data,
in the writing of the report, or in the decision to submit the paper for publicatioPeer reviewe
Î9âTetrahydrocannabinolic acid alleviates collagenâinduced arthritis: Role of PPARÎł and CB1 receptors
Background and Purpose: Î9
-Tetrahydrocannabinolic acid (Î9
-THCA-A), the precur sor of Î9
-THC, is a non-psychotropic phytocannabinoid that shows PPARÎł agonist
activity. Here, we investigated the ability of Î9
-THCA-A to modulate the classic can nabinoid CB1 and CB2 receptors and evaluated its anti-arthritis activity in vitro and
in vivo.
Experimental Approach: Cannabinoid receptors binding and intrinsic activity, as well
as their downstream signalling, were analysed in vitro and in silico. The anti-arthritis
properties of Î9
-THCA-A were studied in human chondrocytes and in the murine
model of collagen-induced arthritis (CIA). Plasma disease biomarkers were identified
by LC-MS/MS based on proteomic and ELISA assays.
Key Results: Functional and docking analyses showed that Î9
-THCA-A can act as an
orthosteric CB1 receptor agonist and also as a positive allosteric modulator in the
presence of CP-55,940. Also, Î9
-THCA-A seemed to be an inverse agonist for CB2
receptors. In vivo, Î9
-THCA-A reduced arthritis in CIA mice, preventing the infiltra tion of inflammatory cells, synovium hyperplasia, and cartilage damage. Furthermore,
Î9
-THCA-A inhibited expression of inflammatory and catabolic genes on knee joints.
The anti-arthritic effect of Î9
-THCA-A was blocked by either SR141716 or
T0070907. Analysis of plasma biomarkers, and determination of cytokines and anti collagen antibodies confirmed that Î9
-THCA-A mediated its activity mainly through
PPARÎł and CB1 receptor pathways.
Conclusion and Implications: Î9
-THCA-A modulates CB1 receptors through the
orthosteric and allosteric binding sites. In addition, Î9
-THCA-A exerts anti-arthritis
activity through CB1 receptors and PPARÎł pathways, highlighting its potential for the
treatment of chronic inflammatory diseases such as rheumatoid arthritis.Instituto de Salud Carlos IIICentre for Industrial Technological DevelopmentUniversidad Complutense de MadridFundaciĂłn Marcelino BotĂnMinisterio de Ciencia e InnovaciĂłnSecretarĂa de Estado de InvestigaciĂłn, Desarrolloe InnovaciĂłnDepto. de BioquĂmica y BiologĂa MolecularFac. de MedicinaTRUEpu
Î9 -Tetrahydrocannabinolic acid alleviates collagen-induced arthritis: Role of PPARÎł and CB1 receptors.
Î9 -Tetrahydrocannabinolic acid (Î9 -THCA-A), the precursor of Î9 -THC, is a non-psychotropic phytocannabinoid that shows PPARÎł agonist activity. Here, we investigated the ability of Î9 -THCA-A to modulate the classic cannabinoid CB1 and CB2 receptors and evaluated its anti-arthritis activity in vitro and in vivo. Cannabinoid receptors binding and intrinsic activity, as well as their downstream signalling, were analysed in vitro and in silico. The anti-arthritis properties of Î9 -THCA-A were studied in human chondrocytes and in the murine model of collagen-induced arthritis (CIA). Plasma disease biomarkers were identified by LC-MS/MS based on proteomic and elisa assays. Functional and docking analyses showed that Î9 -THCA-A can act as an orthosteric CB1 receptor agonist and also as a positive allosteric modulator in the presence of CP-55,940. Also, Î9 -THCA-A seemed to be an inverse agonist for CB2 receptors. In vivo, Î9 -THCA-A reduced arthritis in CIA mice, preventing the infiltration of inflammatory cells, synovium hyperplasia, and cartilage damage. Furthermore, Î9 -THCA-A inhibited expression of inflammatory and catabolic genes on knee joints. The anti-arthritic effect of Î9 -THCA-A was blocked by either SR141716 or T0070907. Analysis of plasma biomarkers, and determination of cytokines and anti-collagen antibodies confirmed that Î9 -THCA-A mediated its activity mainly through PPARÎł and CB1 receptor pathways. Î9 -THCA-A modulates CB1 receptors through the orthosteric and allosteric binding sites. In addition, Î9 -THCA-A exerts anti-arthritis activity through CB1 receptors and PPARÎł pathways, highlighting its potential for the treatment of chronic inflammatory diseases such as rheumatoid arthritis
Targeting nuclear protein TDP-43 by cell division cycle kinase 7 inhibitors: A new therapeutic approach for amyotrophic lateral sclerosis
58 p.-11 fig.-2 tab.Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with no known cure. Aggregates of the nuclear protein TDP-43 have been recognized as a hallmark of proteinopathy in both familial and sporadic cases of ALS. Post-translational modifications of this protein, include hyperphosphorylation, cause disruption of TDP-43 homeostasis and as a consequence, promotion of its neurotoxicity. Among the kinases involved in these changes, cell division cycle kinase 7 (CDC7) plays an important role by directly phosphorylating TDP-43. In the present manuscript the discovery, synthesis, and optimization of a new family of selective and ATP-competitive CDC7 inhibitors based on 6-mercaptopurine scaffold are described. Moreover, we demonstrate the ability of these inhibitors to reduce TDP-43 phosphorylation in both cell cultures and transgenic animal models such as C. elegans and Prp-hTDP43 (A315T) mice. Altogether, the compounds described here may be useful as versatile tools to explore the role of CDC7 in TDP-43 phosphorylation and also as new drug candidates for the future development of ALS therapies.This work was supported by Comunidad de Madrid (grant B2017/BMD-3813), MINECO (grant SAF2016-76693-R and PID2019-105600RB-I00 to A.M., RTI2018-0988885-B-I00 to E.dL. and CTQ2015-66313-R to A.M.R.), ISCii CIBERNED (CB18/05/00040 to A.M., C.G. and A.M.R, and CB06/05/0089 to E.dL.), MECD (FPU14-00204. to E.R.), the National Institutes of Health (R01NS064131 to B.C.K.) and the Department of Veterans Affairs (Merit Review Grants I01BX003755 to B.C.K. and I01BX004044 to N.F.L.), CONICYT-PCI (grant REDES190074 to D.R. and A.M.) and FONDECYT (grant no. 11180604 to D.R.).Peer reviewe
TDP-43 modulation by Tau-Tubulin kinase 1 inhibitors: A new avenue for future amyotrophic lateral sclerosis therapy
68 p.-16 fig.-4 tab.+8 fig. supl.-5 tab. supl.Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease without any effective treatment. Protein TDP-43 is a pathological hallmark of ALS in both sporadic and familiar patients. Post-translational modifications of TDP-43 promote its aggregation in the cytoplasm. Tau-Tubulin kinase (TTBK1) phosphorylates TDP-43 in cellular and animal models; thus, TTBK1 inhibitors emerge as a promising therapeutic strategy for ALS. The design, synthesis, biological evaluation, kinaseâligand complex structure determination, and molecular modeling studies confirmed novel pyrrolopyrimidine derivatives as valuable inhibitors for further development. Moreover, compound 29 revealed good brain penetration in vivo and was able to reduce TDP-43 phosphorylation not only in cell cultures but also in the spinal cord of transgenic TDP-43 mice. A shift to M2 anti-inflammatory microglia was also demonstrated in vivo. Both these activities led to motor neuron preservation in mice, proposing pyrrolopyrimidine 29 as a valuable lead compound for future ALS therapy.This work was supported by Comunidad de Madrid (grant B2017/BMD-3813), the European Social Fund+ (ESF+), MINECO (Grants SAF2016-76693-R to A.M., RTI2018-0988885-B-I00 to E.d.L., and CTQ2015-66313-R to A.M.R.), AIE (RTI2018-099318-B-I00 to D.L., cofunded by the European Regional Development Fund (FEDER)), ISCiii CIBERNED (CB18/05/00040 to A.M., V.P., and A.M.R. and CB06/05/0089 to E.dL.), MECD (FPU16/04466 to V.N.), Cost Action CA15135 âMuTaLigâ (COST-STSM-CA15135-37514 to L.M.G.), FONDECYT (Grant 11180604 to D.R.). V.P. has received financial support through the Postdoctoral Junior Leader Fellowship Program (LCF/BQ/PR18/11640007) from âla Caixaâ Banking Foundation.Peer reviewe
A global point prevalence survey of antimicrobial use in neonatal intensive care units : The no-more-antibiotics and resistance (NO-MAS-R) study
Background: Global assessment of antimicrobial agents prescribed to infants in the neonatal intensive care unit (NICU) may inform antimicrobial stewardship efforts. Methods: We conducted a one-day global point prevalence study of all antimicrobials provided to NICU infants. Demographic, clinical, and microbiologic data were obtained including NICU level, census, birth weight, gestational/chronologic age, diagnoses, antimicrobial therapy (reason for use; length of therapy), antimicrobial stewardship program (ASP), and 30-day in-hospital mortality. Findings: On July 1, 2019, 26% of infants (580/2,265; range, 0-100%; median gestational age, 33 weeks; median birth weight, 1800 g) in 84 NICUs (51, high-income; 33, low-to-middle income) from 29 countries (14, high-income; 15, low-to-middle income) in five continents received >= 1 antimicrobial agent (92%, antibacterial; 19%, antifungal; 4%, antiviral). The most common reasons for antibiotic therapy were "rule-out" sepsis (32%) and "culture-negative" sepsis (16%) with ampicillin (40%), gentamicin (35%), amikacin (19%), vancomycin (15%), and meropenem (9%) used most frequently. For definitive treatment of presumed/confirmed infection, vancomycin (26%), amikacin (20%), and meropenem (16%) were the most prescribed agents. Length of therapy for culture-positive and "culture-negative" infections was 12 days (median; IQR, 8-14) and 7 days (median; IQR, 5-10), respectively. Mortality was 6% (42%, infection-related). An NICU ASP was associated with lower rate of antibiotic utilization (p = 0.02). Interpretation: Global NICU antibiotic use was frequent and prolonged regardless of culture results. NICU-specific ASPs were associated with lower antibiotic utilization rates, suggesting the need for their implementation worldwide