ABSTRACT
The arachidonic acid pathway metabolite 20-hydroxyeicosatetraenoic acid (20-HETE) contributes to ischemia/reperfusion brain injury. Inhibition of 20-HETE formation can protect the developing brain from global ischemia. Here, we examined whether treatment with the 20-HETE synthesis inhibitor N-hydroxy-N-4-butyl-2-methylphenylformamidine (HET0016) can protect the immature brain from traumatic brain injury (TBI). Male rats at postnatal day 9-10 underwent controlled cortical impact followed by intraperitoneal injection with vehicle or HET0016 (1 mg/kg, 5 min and 3 h post-injury). HET0016 decreased the lesion volume by over 50% at 3 days of recovery, and this effect persisted at 30 days as the brain matured. HET0016 decreased peri-lesion gene expression of proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin-1ß [IL-1ß]) at 1 day and increased reparative cytokine (IL-4, IL-10) expression at 3 days. It also partially preserved microglial ramified processes, consistent with less activation. HET0016 decreased contralateral hindlimb foot faults and improved outcome on the novel object recognition memory task 30 days after TBI. In cultured BV2 microglia, HET0016 attenuated the lipopolysaccharide-evoked increase in release of TNF-α. Our data show that HET0016 improves acute and long-term histologic and functional outcomes, in association with an attenuated neuroinflammatory response after contusion of an immature rat brain.
Subject(s)
Amidines/pharmacology , Brain Injuries, Traumatic/drug therapy , Hydroxyeicosatetraenoic Acids/antagonists & inhibitors , Reperfusion Injury/pathology , Animals , Brain/drug effects , Brain/metabolism , Brain/pathology , Brain Injuries, Traumatic/chemically induced , Cytokines/metabolism , Disease Models, Animal , Inflammation/drug therapy , Inflammation/metabolism , Male , Rats, Sprague-Dawley , Reperfusion Injury/chemically inducedABSTRACT
The consequences of therapeutic hypothermia for neonatal hypoxic-ischemic encephalopathy are poorly understood. Adverse effects from suboptimal rewarming could diminish neuroprotection from hypothermia. Therefore, we tested whether rewarming is associated with apoptosis. Piglets underwent hypoxia-asphyxia followed by normothermic or hypothermic recovery at 2 hours. Hypothermic groups were divided into those with no rewarming, rewarming at 0.5 °C/hour, or rewarming at 4 °C/hour. Neurodegeneration at 29 hours was assessed by hematoxylin and eosin staining, TUNEL assay, and immunoblotting for cleaved caspase-3. Rewarmed piglets had more apoptosis in motor cortex than did those that remained hypothermic after hypoxia-asphyxia. Apoptosis in piriform cortex was greater in hypoxic-asphyxic, rewarmed piglets than in naive/sham piglets. Caspase-3 inhibitor suppressed apoptosis with rewarming. Rapidly rewarmed piglets had more caspase-3 cleavage in cerebral cortex than did piglets that remained hypothermic or piglets that were rewarmed slowly. We conclude that rewarming from therapeutic hypothermia can adversely affect the newborn brain by inducing apoptosis through caspase mechanisms.
Subject(s)
Apoptosis , Hypothermia, Induced , Hypoxia-Ischemia, Brain , Infant, Newborn, Diseases , Motor Cortex , Neurons , Animals , Animals, Newborn , Caspases/metabolism , Disease Models, Animal , Humans , Hypoxia-Ischemia, Brain/metabolism , Hypoxia-Ischemia, Brain/pathology , Hypoxia-Ischemia, Brain/therapy , Infant, Newborn , Infant, Newborn, Diseases/metabolism , Infant, Newborn, Diseases/pathology , Infant, Newborn, Diseases/therapy , Motor Cortex/metabolism , Motor Cortex/pathology , Neurons/metabolism , Neurons/pathology , SwineABSTRACT
Nonimmunosuppressant ligands, exemplified by GPI 1046 (1), for the peptidyl-prolyl isomerase FKBP12 have been found to unexpectedly possess powerful neuroprotective and neuroregenerative effects in vitro and in vivo. We have extensively explored the therapeutic utility of FKBP12 ligands based on analogues of proline and pipecolic acid. As part of our ongoing program to explore novel structural classes of FKBP12 ligands, we herein wish to report a new class of FKBP12 ligands containing aza-proline and aza-pipecolic acid analogues. Details of the synthetic studies, together with biological activity will be presented.
Subject(s)
Neuroprotective Agents/chemical synthesis , Neuroprotective Agents/pharmacology , Pipecolic Acids/chemical synthesis , Pipecolic Acids/pharmacology , Proline/analogs & derivatives , Tacrolimus Binding Protein 1A/metabolism , 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/antagonists & inhibitors , Animals , Aza Compounds/chemical synthesis , Aza Compounds/chemistry , Aza Compounds/pharmacology , Corpus Striatum/drug effects , Corpus Striatum/metabolism , Dose-Response Relationship, Drug , Ligands , Male , Mice , Mice, Inbred Strains , Models, Molecular , Neuroprotective Agents/chemistry , Pipecolic Acids/chemistry , Proline/chemical synthesis , Proline/chemistry , Proline/pharmacology , Structure-Activity Relationship , Tacrolimus Binding Protein 1A/chemistry , Tyrosine 3-Monooxygenase/metabolismABSTRACT
As part of our ongoing program to explore novel structural classes of FKBP12 ligands, we herein wish to report a new class of FKBP12 ligands containing chiral bicyclic proline analogues. Details of the synthetic routes, together with preliminary biological activity, will be presented.
Subject(s)
Bridged Bicyclo Compounds/chemical synthesis , Bridged Bicyclo Compounds/pharmacology , Neuroprotective Agents/chemical synthesis , Neuroprotective Agents/pharmacology , Proline/analogs & derivatives , Tacrolimus Binding Protein 1A/drug effects , 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine , Animals , Binding Sites , Dopamine Agents , Indicators and Reagents , Ligands , Male , Mice , Models, Molecular , Molecular Conformation , Parkinson Disease, Secondary/chemically induced , Parkinson Disease, Secondary/drug therapy , Proline/chemical synthesis , Proline/pharmacology , Structure-Activity RelationshipABSTRACT
The recent discovery that small molecule ligands for the peptidyl-prolyl isomerase (PPIase) FKBP12 possess powerful neuroprotective and neuroregenerative properties in vitro and in vivo suggests therapeutic utility for such compounds in neurodegenerative disease. The neurotrophic effects of these compounds are independent of the immunosuppressive pathways by which drugs such as FK506 and rapamycin operate. Previous work by ourselves and other groups exploring the structure-activity relationships (SAR) of small molecules that mimic only the FKBP binding domain portion of FK506 has focused on esters of proline and pipecolic acid. We have explored amide and thioester analogues of these earlier structures and found that they too are extremely potent in promoting recovery of lesioned dopaminergic pathways in a mouse model of Parkinson's disease. Several compounds were shown to be highly effective upon oral administration after lesioning of the dopaminergic pathway, providing further evidence of the potential clinical utility of a variety of structural classes of FKBP12 ligands.
Subject(s)
Amides/chemical synthesis , Nerve Regeneration/drug effects , Pipecolic Acids/chemical synthesis , Proline/analogs & derivatives , Proline/chemical synthesis , Sulfhydryl Compounds/chemical synthesis , Tacrolimus Binding Protein 1A/antagonists & inhibitors , 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine , Administration, Oral , Amides/chemistry , Amides/pharmacology , Animals , Corpus Striatum/enzymology , Corpus Striatum/pathology , Corpus Striatum/ultrastructure , Dopamine Agents , Immunohistochemistry , Ligands , Mice , Molecular Mimicry , Neurites/drug effects , Neuroprotective Agents/chemical synthesis , Neuroprotective Agents/chemistry , Neuroprotective Agents/pharmacology , Parkinson Disease, Secondary/chemically induced , Parkinson Disease, Secondary/drug therapy , Parkinson Disease, Secondary/pathology , Pipecolic Acids/chemistry , Pipecolic Acids/pharmacology , Proline/chemistry , Proline/pharmacology , Structure-Activity Relationship , Substantia Nigra/enzymology , Substantia Nigra/pathology , Substantia Nigra/ultrastructure , Sulfhydryl Compounds/chemistry , Sulfhydryl Compounds/pharmacology , Tacrolimus Binding Protein 1A/chemistry , Tyrosine 3-Monooxygenase/metabolismABSTRACT
The recently discovered small-molecule ligands for the peptidyl and prolyl isomerases (PPIase) of FKBP12 have been shown to possess powerful neuroprotective and neuroregenerative effects. Ketone analogues of the prolyl and pipecolyl esters, which mimic only the FKBP binding domain portion of FK506, are proposed and an efficient synthetic strategy is presented in this report, along with the preliminary results of in vitro and in vivo biological studies.
Subject(s)
Ketones/chemical synthesis , Neuroprotective Agents/chemical synthesis , Pipecolic Acids/chemical synthesis , Proline/analogs & derivatives , Proline/chemical synthesis , Sulfhydryl Compounds/chemical synthesis , Tacrolimus Binding Protein 1A/antagonists & inhibitors , 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine , Animals , Dopamine Agents , Ketones/chemistry , Ketones/pharmacology , Ligands , Mice , Molecular Mimicry , Nerve Regeneration/drug effects , Neurites/drug effects , Neuroprotective Agents/chemistry , Neuroprotective Agents/pharmacology , Parkinson Disease, Secondary/chemically induced , Parkinson Disease, Secondary/drug therapy , Parkinson Disease, Secondary/pathology , Pipecolic Acids/chemistry , Pipecolic Acids/pharmacology , Proline/chemistry , Proline/pharmacology , Structure-Activity Relationship , Sulfhydryl Compounds/chemistry , Sulfhydryl Compounds/pharmacology , Tacrolimus Binding Protein 1A/chemistryABSTRACT
Using simple, inexpensive equipment, we have used solution-phase parallel synthesis to rapidly prepare hundreds of sulfonamide- and urea-containing FKBP inhibitors, resulting in rapid identification of extremely potent compounds in these series.
Subject(s)
Combinatorial Chemistry Techniques/methods , Enzyme Inhibitors/chemical synthesis , Tacrolimus Binding Protein 1A/antagonists & inhibitors , Urea/analogs & derivatives , Drug Design , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Kinetics , Models, Molecular , Molecular Conformation , Pipecolic Acids/chemical synthesis , Pipecolic Acids/pharmacology , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/pharmacology , Urea/chemical synthesis , Urea/pharmacologyABSTRACT
In parallel with our work on solution-phase parallel synthesis of ligands for the rotamase enzyme FKBP12, we herein report a methodology for the solid-phase synthesis of two classes of inhibitor, N-sulfonyl and N-carbamoylprolyl and pipecolyl amides along with their in vitro/in vivo biological results.
