Central Nervous System (CNS) physical injuries, including bacterial or viral infection, can inducechronic neuroinflammation that is believed to persist for the lifetime of an individual. Among the otherinflammatory events, it is recognized that both acute and chronic activation of the complement pathway plays arole in the development of secondary brain injuries by inducing neuronal cell loss and synaptic pruning.Complement over-activation is also firmly implicated in the pathology that underlies the irreversible progressionof multiple sclerosis (MS), a common inflammatory and neurodegenerative disease of the CNS. We hypothesizethat therapeutic inhibition of the complement system and concurrent stimulation of nerve growth may preventCNS tissue damage and slow or even block the progression of MS.Currently, the main obstacle for drug delivery to the CNS is the presence of a selectively permeable blood-brainbarrier (BBB), limiting blood-borne proteins' entryinto the CNS. To overcome this issue, we have recentlydeveloped several potent camelid-derived nanobodies. The first group of nanobodies can inhibit complementactivation, whereas the second group comprises tyrosine kinase receptor TrkB agonists that mimic the action ofbrain-derived neurotrophic factor (BDNF), a growth factor in the brain that promotes neuronal survival, synapticplasticity and neurogenesis. Here, we propose to engineer these nanobodies further to facilitate their crossingof the BBB, thereby gaining the ability to more effectively inhibit the complement cascade and/or stimulate nervegrowth within the CNS.During phase I, bispecific nanobodies will be produced and validated in in vitro cellular functional assays.Therapeutic efficacy will be further validated in a well-characterized murine model of progressive MS, theTheiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD). To this end, mice will betreated with control nanobodies, a complement inhibitor nanobody, TrkB agonistic nanobody, or a combinationof the latter two. Once therapeutic efficacy is confirmed in TMEV-IDD, the camelid-derived nanobodies will behumanized to reduce antigenicity in humans.Statistically significant improvement in treated mice monitored as an impact on disability progression and CNSpathology will be the foundation for a phase II submission. The goals of phase II are 1) revalidate the therapeuticefficacy of humanized antibodies in more extensive experiments, including a detailed analysis of the effect ofsex, age, dose-ranges and delayed treatments, i.e., later than 30 days post-infection, on disability progression,disease pathology and recovery; 2) to establish manufacturing protocols under current Good ManufacturingPractice conditions and; 3) to define the biological response, PK/PD, dose-ranging and toxicology in multiplemodel animals, including toxicology studies in non-human primates.
Public Health Relevance Statement: Narrative: As result of this project, first in class complement inhibiting and neurotropic antibody therapeutics capable of crossing the blood brain barrier for treatment of progressive multiple sclerosis will be obtained.
Project Terms: Adult ; 21+ years old ; Adult Human ; adulthood ; Age ; ages ; Amino Acids ; aminoacid ; Animals ; Antibodies ; Bacterial Infections ; bacteria infection ; bacterial disease ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Blood ; Blood Reticuloendothelial System ; Blood - brain barrier anatomy ; Blood-Brain Barrier ; Hemato-Encephalic Barrier ; bloodbrain barrier ; Brain ; Brain Nervous System ; Encephalon ; Cell Culture Techniques ; cell culture ; Cells ; Cell Body ; Central Nervous System Diseases ; CNS Diseases ; CNS disorder ; Central Nervous System Disorders ; Complement ; Complement Proteins ; Complement 1q ; C1 q ; C1q ; Complement C1q ; Complement Activation ; complement pathway regulation ; Complement Inactivators ; Complement Inhibitors ; Demyelinating Diseases ; Demyelinating Disorders ; Disease ; Disorder ; Engineering ; Exhibits ; Eye diseases ; eye disorder ; ophthalmopathy ; Foundations ; Goals ; Growth ; Generalized Growth ; Tissue Growth ; ontogeny ; Hemolysis ; erythrolysis ; Human ; Modern Man ; In Vitro ; Infection ; Intraperitoneal Injections ; IP injection ; Japanese Population ; Japanese ; Multiple Sclerosis ; Disseminated Sclerosis ; insular sclerosis ; Mus ; Mice ; Mice Mammals ; Murine ; Nerve ; Nerve Degeneration ; Neuron Degeneration ; neural degeneration ; neurodegeneration ; neurodegenerative ; neurological degeneration ; neuronal degeneration ; Ophthalmology ; Pathology ; Patients ; Peptides ; Play ; Production ; Proteins ; Thrombotic Thrombocytopenic Purpura ; Moschkowitz Disease ; Thrombotic Thrombopenic Purpura ; Rabies virus ; Role ; social role ; medical schools ; medical college ; school of medicine ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Testing ; Tissues ; Body Tissues ; Toxicology ; Virus Diseases ; Viral Diseases ; viral infection ; virus infection ; virus-induced disease ; Measures ; Neurotrophic Tyrosine Kinase Receptor Type 2 ; BDNF Receptor ; BDNF/NT-3 Growth Factors Receptor ; Brain-Derived Neurotrophic Factor Receptor ; GP145-TRKB ; NTRK2 Receptor ; TRKB Tyrosine Kinase ; trkB Receptor ; trkB(gp145) Protein ; Drug Delivery Systems ; Drug Delivery ; Brain-Derived Neurotrophic Factor ; BDNF ; Catalytic Antibodies ; abzyme ; Injury ; injuries ; base ; improved ; Acute ; Chronic ; Clinical ; Phase ; Variant ; Variation ; Biological ; Receptor Protein-Tyrosine Kinases ; PTK Receptors ; Receptor Tyrosine Kinase Gene ; Transmembrane Receptor Protein Tyrosine Kinase ; Tyrosine Kinase Linked Receptors ; Tyrosine Kinase Receptors ; TMEV ; Mouse Polioviruses ; Murine Polioviruses ; Theiler Murine Encephalomyelitis Virus ; Theiler's Virus ; Theiler's encephalomyelitis virus ; Bispecific Antibodies ; Bi-specific antibodies ; Bifunctional Antibodies ; bsAb ; disability ; Individual ; nonhuman primate ; non-human primate ; Recovery ; Agonist ; Brain Injuries ; Acquired brain injury ; brain damage ; brain-injured ; Antibody Therapy ; antibody based therapies ; antibody treatment ; antibody-based therapeutics ; antibody-based treatment ; Collaborations ; Therapeutic ; Inflammatory ; Reporter ; Event ; Protocol ; Protocols documentation ; Degenerative Neurologic Diseases ; Degenerative Neurologic Disorders ; Nervous System Degenerative Diseases ; Neural Degenerative Diseases ; Neural degenerative Disorders ; Neurodegenerative Diseases ; Neurologic Degenerative Conditions ; degenerative diseases of motor and sensory neurons ; degenerative neurological diseases ; neurodegenerative illness ; Neurodegenerative Disorders ; brain cell ; nerve cell death ; nerve cell loss ; neuron cell death ; neuron cell loss ; neuron death ; neuronal cell death ; neuronal cell loss ; neuronal death ; neuronal loss ; neuron loss ; Animal Models and Related Studies ; model of animal ; model organism ; Animal Model ; treatment planning ; Disease Outcome ; Agreement ; disorder model ; Disease model ; Position ; Positioning Attribute ; neurogenesis ; response ; CNS Nervous System ; Central Nervous System ; Neuraxis ; Molecular Interaction ; Binding ; Pharmaceutical Agent ; Pharmaceuticals ; Pharmacological Substance ; Pharmacologic Substance ; preventing ; prevent ; complement pathway ; Dose ; Data ; in vivo ; Rodent Model ; Validation ; Pathologic ; Monitor ; Molecular ; sex ; Development ; developmental ; neuroinflammation ; neuroinflammatory ; Treatment Efficacy ; intervention efficacy ; therapeutic efficacy ; therapy efficacy ; Outcome ; Synaptic plasticity ; N-terminal ; NH2-terminal ; C-terminal ; Therapeutic antibodies ; treatment effect ; neurotropic ; mouse model ; murine model ; neuronal survival ; humanized antibody ; good laboratory practice ; complement system ; nanobodies ; nanobody ; sdAb ; single domain antibodies ; behavioral study ; behavior study ; murine antibody ; Growth Factor ; Growth Agents ; Growth Substances ; Proteins Growth Factors ; experimental study ; experiment ; experimental research ; synaptic pruning ; pharmacokinetics and pharmacodynamics ; PK/PD ; blood-brain barrier penetration ; BBB penetration ; bloodbrain barrier penetration ; blood-brain barrier permeabilization ; BBB permeabilization ; BBB permeable ; blood-brain barrier permeable ; bloodbrain barrier permeabilization ; bloodbrain barrier permeable ; blood-brain barrier crossing ; BBB crossing ; bloodbrain barrier crossing ; effectiveness testing ;
