SBIR-STTR Award

Monoclonal antibodies (MAb) are potential new therapeutics for many brain diseases, including Alzheimer's disease (AD), Parkinson's disease, mad cow disease, West Nile encephalitis, neuro-AIDS, brain injury, brain cancer, or multiple sclerosis. In almost all cases, it is necessary that the MAb therapeutic that is administered into the blood be able to access target sites within the brain. However, MAb's are large molecule drugs that do not cross the blood-brain barrier (BBB). The BBB problem prevents the brain drug development of antibody drugs. The proposed research will develop a new technology for antibody drug delivery to brain, which could also be used for other organs. This work is based on the genetic engineering of a fusion protein comprised of 2 antibodies. One antibody is the therapeutic antibody, and the other antibody is a drug delivery system. The prototype of this new technology is novel fusion protein, which is created by fusion of a single chain Fv (ScFv) antibody to the carboxyl terminus of both heavy chains of a genetically engineered MAb against the human insulin receptor (HIR). The MAb against the HIR does penetrate brain via transport across the BBB on the endogenous insulin receptor. Feasibility studies show the fusion protein retains its bi-functional properties following genetic engineering and transient expression in COS cells: the fusion antibody both binds the HIR, to enable transport across the BBB in vivo, and binds Abeta plaque, to cause amyloid disaggregation in AD transgenic mouse brain. The purpose of these Phase I studies is (1) genetically engineer a tandem vector expressing the hetero-tetrameric fusion protein, (2) to engineer a permanently transfected host cell line that expresses high levels of the fusion antibody in serum free medium, (3) to biochemically and functionally characterize the fusion antibody, and (4) determine the plasma pharmacokinetics and brain uptake of the fusion protein in the adult Rhesus monkey. If successful, this research will develop a new technology for the drug delivery of antibody therapeutics.

Public Health Relevance:
Monoclonal antibodies are powerful new therapeutic products of biotechnology. Antibody drugs could be applied to many serious brain disorders, such as Alzheimer's disease, Parkinson's disease, mad cow disease, West Nile encephalitis, neuro-AIDS, brain injury, brain cancer, or multiple sclerosis. However, antibody drugs cannot be developed for these disorders, because the antibody drugs do not cross the blood-brain barrier (BBB). The present research will develop a new technology for the drug delivery of antibody drugs for the brain, which could be applied to other organs.

Public Health Relevance:
This Public Health Relevance is not available.

Thesaurus Terms:
There Are No Thesaurus Terms On File For This Project.

Monoclonal antibodies (MAb) are potential new therapeutics for many brain diseases, including Alzheimer's disease (AD), Parkinson's disease, mad cow disease, West Nile encephalitis, neuro- AIDS, brain injury, brain cancer, or multiple sclerosis. In almost all cases, it is necessary that the MAb therapeutic that is administered into the blood be able to access target sites within the brain. However, MAb's are large molecule drugs that do not cross the blood-brain barrier (BBB). The BBB problem prevents the brain drug development of antibody drugs. The proposed research will develop a new technology for antibody drug delivery to brain, which could also be used for other organs, and the new technology will be applied to AD. This work is based on the genetic engineering of a fusion protein comprised of 2 antibodies. One antibody is the therapeutic antibody against the Abeta amyloid peptide of AD, and the other antibody is a drug delivery system, which is directed at an endogenous transporter on the human BBB. The Phase I studies accomplished the following: (1) genetic engineering of a tandem vector expressing the hetero-tetrameric fusion protein, (2) cloning of a permanently transfected host cell line that expresses high levels of the fusion antibody in serum free medium, (3) biochemical and functional characterizion of the fusion antibody, and (4) determination of the plasma pharmacokinetics (PK) and brain uptake of the fusion protein in the adult Rhesus monkey. The phase II studies will accomplish the following: (1) growth of the host cell line in a 50L bioreactor, followed by 3-column downstream processing that can be replicated in a GMP lab;(2) biochemical analysis of the fusion protein with over 15 analytical tests;(3) dose finding PK and toxicity study in Rhesus monkeys. These studies will enable future submission of an IND for human testing of this new fusion protein for AD.

Public Health Relevance:
Monoclonal antibodies are powerful new therapeutic products of biotechnology. Antibody drugs could be applied to many serious brain disorders, such as Alzheimer's disease (AD), Parkinson's disease, mad cow disease, West Nile encephalitis, neuro-AIDS, brain injury, brain cancer, or multiple sclerosis. However, antibody drugs cannot be developed for these disorders, because the antibody drugs do not cross the blood-brain barrier (BBB). The present research will develop a new technology for the drug delivery of antibody drugs for the brain, which could be applied to diseases such as AD.

Thesaurus Terms:
21+ Years Old;Aids;Accounting;Acquired Immune Deficiency;Acquired Immune Deficiency Syndrome;Acquired Immuno-Deficiency Syndrome;Acquired Immunodeficiency Syndrome;Acquired Brain Injury;Active Immunization;Adult;Affinity;Affinity Chromatography;Alzheimer;Alzheimer Beta-Protein;Alzheimer Disease;Alzheimer Sclerosis;Alzheimer Syndrome;Alzheimer's;Alzheimer's Disease;Alzheimer's Amyloid;Alzheimers Dementia;Alzheimers Disease;Amentia;Amyloid Alzeheimer's Dementia Amyloid Protein;Amyloid Beta-Peptide;Amyloid Fibril Protein (Alzheimer's);Amyloid Plaques;Amyloid Protein A4;Amyloid Beta-Protein;Amyloid Beta-Protein, Alzheimer's;Animals;Anions;Antibodies;Appearance;Assay;Astroprotein;Autopsy;Berlin Blue;Binding;Binding (Molecular Function);Bioassay;Biochemical;Biologic Assays;Biological Assay;Biomedical Engineering;Bioreactors;Biotechnology;Biotechnology, Genetic Engineering;Bleeding;Blood;Blood - Brain Barrier Anatomy;Blood Plasma;Blood-Brain Barrier;Blotting, Western;Body Weight;Bovine Spongiform Encephalopathy;Brain;Brain Diseases;Brain Disorders;Brain Injuries;Cho Cells;Cancer Of Brain;Carbohydrates;Cations;Cell Line;Cell Lines, Strains;Cellline;Cells;Cerebrum;Chimera Protein;Chimeric Proteins;Chinese Hamster;Chinese Hamster Ovary Cell;Chromatography;Chromatography / Separation Science;Chromatography, Affinity;Chromatography, High Performance Liquid;Chromatography, High Pressure Liquid;Chromatography, High Speed Liquid;Clinical Chemistry Tests;Cloning;Complex;Culture Media, Serum-Free;Dna;Dementia;Dementia, Alzheimer Type;Dementia, Primary Senile Degenerative;Dementia, Senile;Deoxyribonucleic Acid;Development;Disease;Disorder;Dose;Drug Delivery;Drug Delivery Systems;Drug Kinetics;Drug Targeting;Drug Targetings;Drugs;Elisa;Egypt 101 Virus;Electrofocusing;Encephalitis;Encephalitis, Bovine Spongiform;Encephalon;Encephalon Diseases;Encephalons;Encephalopathy, Bovine Spongiform;Endotoxins;Engineering;Engineerings;Enzyme-Linked Immunosorbent Assay;Exclusion;Fc Receptor, Neonatal;Fcrn;Fcrn Neonatal Transfer Protein;Fcrn Protein, Human;Filtration;Fluorescence Microscopy;Fractionation, Filtration;Fractionation, Isoelectric Focusing;Fusion Protein;Future;Gfa-Protein;Gfap;Gamma Globulin, 7s;Generalized Growth;Genetic Engineering;Genital System, Female, Ovary;Glial Fibrillary Acid Protein;Glial Fibrillary Acidic Protein;Glial Intermediate Filament Protein;Growth;H And E;Hplc;Hemato-Encephalic Barrier;Hematoxylin And Eosin;Hematoxylin And Eosin Staining Method;Hemorrhage;High Pressure Liquid Chromatography;Histocytochemistry;Human;Human, Adult;Human, General;Insr;Insr Protein, Human;Itx;Idiopathic Parkinson Disease;Igg;Immunoglobulin G;Immunologic Deficiency Syndrome, Acquired;Immunologically Directed Therapy;Immunotherapy;Inflammation, Brain;Injection Of Therapeutic Agent;Injections;Insulin Receptor;Insulin Receptor Protein-Tyrosine Kinase;Insulin-Dependent Tyrosine Protein Kinase;Intracranial Cns Disorders;Intracranial Central Nervous System Disorders;Isoelectric Focusing;Lewy Body Parkinson Disease;Mab Therapeutics;Ms (Multiple Sclerosis);Macaca Mulatta;Mad Cow Disease;Malignant Tumor Of The Brain;Malignant Neoplasm Of Brain;Mammals, Primates;Man (Taxonomy);Man, Modern;Mediating;Medication;Microscopy, Fluorescence;Microscopy, Light, Fluorescence;Moab, Clinical Treatment;Molecular Biology, Genetic Engineering;Molecular Interaction;Monoclonal Antibodies;Multiple Sclerosis;Nervous System, Brain;Neuritic Plaques;Organ;Organ Weight;Ovary;Paralysis Agitans;Parkinson;Parkinson Disease;Parkinson's;Parkinson's Disease;Parkinsons Disease;Passive Immunization;Peptide Fingerprinting;Peptide Mapping;Peptides;Perfusion;Peripheral;Pharmaceutic Preparations;Pharmaceutical Preparations;Pharmacokinetics;Phase;Plasma;Primary Parkinsonism;Primary Senile Degenerative Dementia;Primates;Process;Production;Programs (Pt);Programs [publication Type];Progress Reports;Protein Binding;Protein-Free Media;Proteins;Prussian Blue;Publications;Receptor Protein;Recombinant Dna Technology;Recombinant Fusion Proteins;Reference Standards;Relative;Relative (Related Person);Reporting;Research;Reticuloendothelial System, Blood;Reticuloendothelial System, Serum, Plasma;Rhesus;Rhesus Macaque;Rhesus Monkey;Sbir;Sbirs (R43/44);Sds Page;Sds Polyacrylamide Gel Electrophoresis;Scientific Publication;Sclerosis, Disseminated;Senile Plaques;Serum, Plasma;Serum-Free Culture Media;Serum-Free Media;Site;Small Business Innovation Research;Small Business Innovation Research Grant;Sodium Dodecyl Sulfate-Page;Sodium Dodecylsulfate-Polyacrylamide Gel Electrophoresis;Symptoms;Technology;Temperature;Testing;Therapeutic Monoclonal Antibodies;Therapeutic Antibodies;Time;Tissue Growth;Tissue Stains;Toxic Effect;Toxicities;Toxicology;Transgenic Mice;Urinary System, Urine;Urine;Wnv;West Nile;West Nile Virus;Western Blotting;Western Blottings;Western Immunoblotting;Work;A Beta Peptide;Abeta;Adult Human (21+);Affinity Chromatography;Affinity Purification;Amyloid Beta;Amyloid Beta Plaque;Amyloid Peptide;Amyloid-B Plaque;Amyloid-B Protein;Antibody;Base;Beta Amyloid Fibril;Bioengineering;Bioengineering /Biomedical Engineering;Bioengineering/Biomedical Engineering;Blood Brain Barrier;Blood Loss;Brain Damage;Brain Injury;Brain Lesion (From Injury);Brain Tissue;Cored Plaque;Cultured Cell Line;Dementia Of The Alzheimer Type;Diffuse Plaque;Disease /Disorder;Disease/Disorder;Drug /Agent;Drug Development;Drug/Agent;Enzyme Linked Immunosorbent Assay;Ferric Ferrocyanide;Ferrihexacyanoferrate;Ferrotsin;Fluorescence Microscopy;Fluoro Jade;Gene Product;High Performance Liquid Chromatography;Histochemistry;Histochemistry /Cytochemistry;Histochemistry/Cytochemistry;Human Insr Protein;Immune Therapy;Immunocytochemistry;Insular Sclerosis;Insulin Receptor, Human;Manufacturing Process;Molecular Trojan Horse;Molecular Trojan Horse Technology;Monoclonal Antibody;Nano;Necropsy;Neonatal Fc Receptor;Neuropathology;New Technology;New Therapeutics;Next Generation Therapeutics;Novel Therapeutics;Ontogeny;Passive Immunization;Phase 1 Study;Phase 2 Study;Postmortem;Prevent;Preventing;Primary Degenerative Dementia;Programs;Protein Blotting;Receptor;Receptor Binding;Senile Dementia Of The Alzheimer Type;Soluble Amyloid Precursor Protein;Uptake;Vector