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1.
Nature ; 560(7718): 372-376, 2018 08.
Artículo en Inglés | MEDLINE | ID: mdl-30046109

RESUMEN

Regeneration of myelin is mediated by oligodendrocyte progenitor cells-an abundant stem cell population in the central nervous system (CNS) and the principal source of new myelinating oligodendrocytes. Loss of myelin-producing oligodendrocytes in the CNS underlies a number of neurological diseases, including multiple sclerosis and diverse genetic diseases1-3. High-throughput chemical screening approaches have been used to identify small molecules that stimulate the formation of oligodendrocytes from oligodendrocyte progenitor cells and functionally enhance remyelination in vivo4-10. Here we show that a wide range of these pro-myelinating small molecules function not through their canonical targets but by directly inhibiting CYP51, TM7SF2, or EBP, a narrow range of enzymes within the cholesterol biosynthesis pathway. Subsequent accumulation of the 8,9-unsaturated sterol substrates of these enzymes is a key mechanistic node that promotes oligodendrocyte formation, as 8,9-unsaturated sterols are effective when supplied to oligodendrocyte progenitor cells in purified form whereas analogous sterols that lack this structural feature have no effect. Collectively, our results define a unifying sterol-based mechanism of action for most known small-molecule enhancers of oligodendrocyte formation and highlight specific targets to propel the development of optimal remyelinating therapeutics.


Asunto(s)
Vaina de Mielina/metabolismo , Oligodendroglía/citología , Oligodendroglía/metabolismo , Remielinización , Esteroles/química , Esteroles/metabolismo , Inhibidores de 14 alfa Desmetilasa/farmacología , Animales , Colesterol/biosíntesis , Células HEK293 , Ensayos Analíticos de Alto Rendimiento , Humanos , Imidazoles/farmacología , Masculino , Proteínas de la Membrana/antagonistas & inhibidores , Ratones , Ratones Endogámicos C57BL , Esclerosis Múltiple , Oligodendroglía/efectos de los fármacos , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/antagonistas & inhibidores , Remielinización/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos , Esferoides Celulares/metabolismo , Médula Espinal/efectos de los fármacos , Médula Espinal/patología , Esteroide Isomerasas/antagonistas & inhibidores , Esterol 14-Desmetilasa/metabolismo , Especificidad por Sustrato
2.
Chemistry ; 29(53): e202301506, 2023 Sep 21.
Artículo en Inglés | MEDLINE | ID: mdl-37415318

RESUMEN

The reaction of [(L)MnII ]2+ (L = neutral polypyridine ligand framework) in the presence of mCPBA (mCPBA = m-Chloroperoxybenzoic acid) generates a putative MnV =O species at RT. The proposed MnV =O species is capable of performing the aromatic hydroxylation of Cl-benzoic acid derived from mCPBA to give [(L)MnIII (m-Cl-salicylate)]+ , which in the presence of excess mCPBA generates a metastable [(L)MnV (O)(m-Cl-salicylate)]+ , characterized by UV/Vis absorption, EPR, resonance Raman spectroscopy, and ESI-MS studies. The current study highlights the fact that [(L)MnIII (m-Cl-salicylate)]+ formation may not be a dead end for catalysis. Further, a plausible mechanism has been proposed for the formation of [(L)MnV (O)-m-Cl-salicylate)]+ from [(L)MnIII (m-Cl-salicylate)]+ . The characterized transient [(L)MnV (O)-m-Cl-salicylate)]+ reported in the current work exhibits high reactivity for oxygen atom transfer reactions, supported by the electrophilic character depicted from Hammett studies using a series of para-substituted thioanisoles. The unprecedented study starting from a non-heme neutral polypyridine ligand framework paves a path for mimicking the natural active site of photosystem II under ambient conditions. Finally, evaluating the intracellular effect of Mn(II) complexes revealed an enhanced intracellular ROS and mitochondrial dysfunction to prevent the proliferation of hepatocellular carcinoma and breast cancer cells.


Asunto(s)
Manganeso , Neoplasias , Manganeso/química , Ligandos , Clorobenzoatos
3.
Chem Commun (Camb) ; 60(14): 1928-1931, 2024 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-38268331

RESUMEN

Developing new chemotherapeutics that are structurally and mechanistically unique is needed due to the rapid rise of the cancer incidence across the globe. Here, we report the identification of irreversible, thiol-reactive diazepam derivatives as GPX4 modifiers and nanomolar inducers of ferroptosis in liver cancer cells.


Asunto(s)
Ferroptosis , Neoplasias Hepáticas , Humanos , Línea Celular , Diazepam , Compuestos de Sulfhidrilo , Neoplasias Hepáticas/tratamiento farmacológico
4.
iScience ; 27(6): 109899, 2024 Jun 21.
Artículo en Inglés | MEDLINE | ID: mdl-38799569

RESUMEN

The emergence of multidrug resistance in cancer cells necessitates the development of new therapeutic modalities. One way cancer cells orchestrate energy metabolism and redox homeostasis is through overloaded iron pools directed by iron regulatory proteins, including transferrin. Here, we demonstrate that targeting redox homeostasis using nitrogen-based heterocyclic iron chelators and their iron complexes efficiently prevents the proliferation of liver cancer cells (EC50: 340 nM for IITK4003) and liver cancer 3D spheroids. These iron complexes generate highly reactive Fe(IV)=O species and accumulate lipid peroxides to promote oxidative stress in cells that impair mitochondrial function. Subsequent leakage of mitochondrial cytochrome c activates the caspase cascade to trigger the intrinsic apoptosis pathway in cancer cells. This strategy could be applied to leverage the inherent iron overload in cancer cells to selectively promote intrinsic cellular apoptosis for the development of unique iron-complex-based anticancer therapeutics.

5.
ACS Chem Biol ; 17(8): 2188-2200, 2022 08 19.
Artículo en Inglés | MEDLINE | ID: mdl-35833657

RESUMEN

Regeneration of myelin in the central nervous system is being pursued as a potential therapeutic approach for multiple sclerosis. Several labs have reported small molecules that promote oligodendrocyte formation and remyelination in vivo. Recently, we reported that many such molecules function by inhibiting a narrow window of enzymes in the cholesterol biosynthesis pathway. Here we describe a new high-throughput screen of 1,836 bioactive molecules and a thorough re-analysis of more than 60 molecules previously identified as promoting oligodendrocyte formation from human, rat, or mouse oligodendrocyte progenitor cells. These studies highlight that an overwhelming fraction of validated screening hits, including several molecules being evaluated clinically for remyelination, inhibit cholesterol pathway enzymes like emopamil-binding protein (EBP). To rationalize these findings, we suggest a model that relies on the high druggability of sterol-metabolizing enzymes and the ability of cationic amphiphiles to mimic the transition state of EBP. These studies further establish cholesterol pathway inhibition as a dominant mechanism among screening hits that enhance human, rat, or mouse oligodendrocyte formation.


Asunto(s)
Remielinización , Roedores , Animales , Diferenciación Celular , Colesterol/metabolismo , Humanos , Ratones , Oligodendroglía/metabolismo , Ratas
6.
Chem Commun (Camb) ; 58(83): 11669-11672, 2022 Oct 18.
Artículo en Inglés | MEDLINE | ID: mdl-36172810

RESUMEN

Antimicrobial resistance (AMR) is a global health concern. Targetting AMR, we present an in situ lactonization mechanism generating 4-nitroisobenzofuran-1(3H)-one (IITK2020), an exclusive S. aureus inhibitor at 2-4 µg mL-1 MIC including multidrug-resistant S. aureus clinical strains, that prevents peptidoglycan biosynthesis.


Asunto(s)
Antiinfecciosos , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Humanos , Pruebas de Sensibilidad Microbiana , Peptidoglicano , Staphylococcus aureus
7.
ACS Chem Biol ; 16(7): 1288-1297, 2021 07 16.
Artículo en Inglés | MEDLINE | ID: mdl-34232635

RESUMEN

Inducing the formation of new oligodendrocytes from oligodendrocyte progenitor cells (OPCs) represents a potential approach to repairing the loss of myelin observed in multiple sclerosis and other diseases. Recently, we demonstrated that accumulation of specific cholesterol precursors, 8,9-unsaturated sterols, is a dominant mechanism by which dozens of small molecules enhance oligodendrocyte formation. Here, we evaluated a library of 56 sterols and steroids to evaluate whether other classes of bioactive sterol derivatives may also influence mouse oligodendrocyte precursor cell (OPC) differentiation or survival. From this library, we identified U-73343 as a potent enhancer of oligodendrocyte formation that induces 8,9-unsaturated sterol accumulation by inhibition of the cholesterol biosynthesis enzyme sterol 14-reductase. In contrast, we found that mouse OPCs are remarkably vulnerable to treatment with the glycosterol OSW-1, an oxysterol-binding protein (OSBP) modulator that induces Golgi stress and OPC death in the low picomolar range. A subsequent small-molecule suppressor screen identified mTOR signaling as a key effector pathway mediating OSW-1's cytotoxic effects in mouse OPCs. Finally, evaluation of a panel of ER and Golgi stress-inducing small molecules revealed that mouse OPCs are highly sensitive to these perturbations, more so than closely related neural progenitor cells. Together, these studies highlight the wide-ranging influence of sterols and steroids on OPC cell fate, with 8,9-unsaturated sterols positively enhancing differentiation to oligodendrocytes and OSW-1 able to induce lethal Golgi stress with remarkable potency.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Células Precursoras de Oligodendrocitos/efectos de los fármacos , Esteroles/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Colestenonas/farmacología , Colestenonas/toxicidad , Evaluación Preclínica de Medicamentos , Retículo Endoplásmico/efectos de los fármacos , Estrés del Retículo Endoplásmico/efectos de los fármacos , Estrenos/farmacología , Aparato de Golgi/efectos de los fármacos , Células HeLa , Humanos , Ratones , Células Precursoras de Oligodendrocitos/metabolismo , Oligodendroglía/metabolismo , Pirrolidinonas/farmacología , Saponinas/farmacología , Saponinas/toxicidad , Bibliotecas de Moléculas Pequeñas/farmacología , Bibliotecas de Moléculas Pequeñas/toxicidad , Esteroles/toxicidad
8.
Cell Chem Biol ; 28(6): 866-875.e5, 2021 06 17.
Artículo en Inglés | MEDLINE | ID: mdl-33636107

RESUMEN

Small molecules that promote the formation of new myelinating oligodendrocytes from oligodendrocyte progenitor cells (OPCs) are potential therapeutics for demyelinating diseases. We recently established inhibition of specific cholesterol biosynthesis enzymes and resulting accumulation of 8,9-unsaturated sterols as a unifying mechanism through which many such molecules act. To identify more potent sterol enhancers of oligodendrocyte formation, we synthesized a collection of 8,9-unsaturated sterol derivatives and found that 24,25-epoxylanosterol potently promoted oligodendrocyte formation. In OPCs, 24,25-epoxylanosterol was metabolized to 24,25-epoxycholesterol via the epoxycholesterol shunt pathway. Increasing flux through the epoxycholesterol shunt using genetic manipulation or small-molecule inhibition of lanosterol synthase (LSS) increased endogenous 24,25-epoxycholesterol levels and OPC differentiation. Notably, exogenously supplied 24,25-epoxycholesterol promoted oligodendrocyte formation despite lacking an 8,9-unsaturation. This work highlights epoxycholesterol shunt usage, controlled by inhibitors of LSS, as a target to promote oligodendrocyte formation. Additionally, sterols beyond the 8,9-unsaturated sterols, including 24,25-epoxycholesterol, drive oligodendrocyte formation.


Asunto(s)
Colesterol/análogos & derivados , Transferasas Intramoleculares/metabolismo , Oligodendroglía/metabolismo , Animales , Células Cultivadas , Colesterol/biosíntesis , Colesterol/química , Masculino , Ratones , Oligodendroglía/citología
9.
Cell Chem Biol ; 26(4): 593-599.e4, 2019 04 18.
Artículo en Inglés | MEDLINE | ID: mdl-30773481

RESUMEN

Small molecules that promote oligodendrocyte formation have been identified in "drug repurposing" screens to nominate candidate therapeutics for diseases in which myelin is lost, including multiple sclerosis. We recently reported that many such molecules enhance oligodendrocyte formation not by their canonical targets but by inhibiting a narrow range of enzymes in cholesterol biosynthesis. Here we identify enhancers of oligodendrocyte formation obtained by screening a structurally diverse library of 10,000 small molecules. Identification of the cellular targets of these validated hits revealed a majority inhibited the cholesterol biosynthesis enzymes CYP51, TM7SF2, or EBP. In addition, evaluation of analogs led to identification of CW3388, a potent EBP-inhibiting enhancer of oligodendrocyte formation poised for further optimization.


Asunto(s)
Inhibidores de 14 alfa Desmetilasa/farmacología , Oligodendroglía/efectos de los fármacos , Oxidorreductasas/antagonistas & inhibidores , Bibliotecas de Moléculas Pequeñas/farmacología , Esteroide Isomerasas/antagonistas & inhibidores , Inhibidores de 14 alfa Desmetilasa/química , Animales , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Ratones , Oligodendroglía/citología , Oligodendroglía/metabolismo , Oxidorreductasas/metabolismo , Bibliotecas de Moléculas Pequeñas/química , Esteroide Isomerasas/metabolismo
11.
ACS Chem Biol ; 12(8): 2124-2131, 2017 08 18.
Artículo en Inglés | MEDLINE | ID: mdl-28613814

RESUMEN

Resurgent interest in covalent target engagement in drug discovery has demonstrated that small molecules containing weakly reactive electrophiles can be safe and effective therapies. Several recently FDA-approved drugs feature an acrylamide functionality to selectively engage cysteine side chains of kinases (Ibrutinib, Afatinib, and Neratinib). Additional electrophilic functionalities whose reactivity is compatible with highly selective target engagement and in vivo application could open new avenues in covalent small molecule discovery. Here, we report the synthesis and evaluation of a library of small molecules containing the 2-chloropropionamide functionality, which we demonstrate is less reactive than typical acrylamide electrophiles. Although many library members do not appear to label proteins in cells, we identified S-CW3554 as selectively labeling protein disulfide isomerase and inhibiting its enzymatic activity. Subsequent profiling of the library against five diverse cancer cell lines showed unique cytotoxicity for S-CW3554 in cells derived from multiple myeloma, a cancer recently reported to be sensitive to PDI inhibition. Our novel PDI inhibitor highlights the potential of 2-chloropropionamides as weak and stereochemically tunable electrophiles for covalent drug discovery.


Asunto(s)
Amidas/farmacología , Sistemas de Liberación de Medicamentos , Diseño de Fármacos , Proteína Disulfuro Isomerasas/metabolismo , Bibliotecas de Moléculas Pequeñas/química , Triptófano/análogos & derivados , Amidas/química , Elementos de Respuesta Antioxidante , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Células HEK293 , Humanos , Estructura Molecular , Mieloma Múltiple/tratamiento farmacológico , Bibliotecas de Moléculas Pequeñas/farmacología , Triptófano/química , Triptófano/farmacología
12.
PLoS One ; 12(11): e0187991, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29155853

RESUMEN

Glutaredoxin (Grx1) is a ubiquitously expressed thiol-disulfide oxidoreductase that specifically catalyzes reduction of S-glutathionylated substrates. Grx1 is known to be a key regulator of pro-inflammatory signaling, and Grx1 silencing inhibits inflammation in inflammatory disease models. Therefore, we anticipate that inhibition of Grx1 could be an anti-inflammatory therapeutic strategy. We used a rapid screening approach to test 504 novel electrophilic compounds for inhibition of Grx1, which has a highly reactive active-site cysteine residue (pKa 3.5). From this chemical library a chloroacetamido compound, CWR-J02, was identified as a potential lead compound to be characterized. CWR-J02 inhibited isolated Grx1 with an IC50 value of 32 µM in the presence of 1 mM glutathione. Mass spectrometric analysis documented preferential adduction of CWR-J02 to the active site Cys-22 of Grx1, and molecular dynamics simulation identified a potential non-covalent binding site. Treatment of the BV2 microglial cell line with CWR-J02 led to inhibition of intracellular Grx1 activity with an IC50 value (37 µM). CWR-J02 treatment decreased lipopolysaccharide-induced inflammatory gene transcription in the microglial cells in a parallel concentration-dependent manner, documenting the anti-inflammatory potential of CWR-J02. Exploiting the alkyne moiety of CWR-J02, we used click chemistry to link biotin azide to CWR-J02-adducted proteins, isolating them with streptavidin beads. Tandem mass spectrometric analysis identified many CWR-J02-reactive proteins, including Grx1 and several mediators of inflammatory activation. Taken together, these data identify CWR-J02 as an intracellularly effective Grx1 inhibitor that may elicit its anti-inflammatory action in a synergistic manner by also disabling other pro-inflammatory mediators. The CWR-J02 molecule provides a starting point for developing more selective Grx1 inhibitors and anti-inflammatory agents for therapeutic development.


Asunto(s)
Acetanilidas/farmacología , Antiinflamatorios/farmacología , Glutarredoxinas/antagonistas & inhibidores , Microglía/efectos de los fármacos , Ácidos Ftálicos/farmacología , Acetanilidas/síntesis química , Secuencia de Aminoácidos , Animales , Antiinflamatorios/síntesis química , Sitios de Unión , Biotina/química , Línea Celular , Química Clic , Expresión Génica , Glutarredoxinas/química , Glutarredoxinas/genética , Glutarredoxinas/metabolismo , Ensayos Analíticos de Alto Rendimiento , Cinética , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/farmacología , Ratones , Microglía/citología , Microglía/metabolismo , Simulación de Dinámica Molecular , Ácidos Ftálicos/síntesis química , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Estreptavidina/química , Termodinámica
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