SBIR-STTR Award

Type II Mucopolysaccharidosis (MPS), also known as Hunter's syndrome, is an genetic disease that affects the lysosomal enzyme, iduronate-2-sulfatase (IDS). Consequently, patients with MPS-II develop extensive lysosomal storage product accumulation in tissues, including the brain. Children develop multiple brain disorders including mental retardation and hydrocephalus. The current therapy for MPS-II is Enzyme Replacement Therapy (ERT) with the recombinant human protein. However, ERT does not treat the brain of MPS-II, because IDS does not cross the blood-brain barrier (BBB). The present work will re-engineer human IDS to enable transport across the BBB, using molecular Trojan horse technology. A molecular Trojan horse is a genetically engineered peptidomimetic monoclonal antibody (MAb) against an endogenous BBB peptide receptor, such as the human insulin receptor (HIR). The human IDS is fused to the heavy chain of the HIRMAb to create a new chemical entity, called the HIRMAb-IDS fusion protein. Preliminary studies show the HIRMAb-IDS fusion protein retains high IDS enzyme activity and high binding for the HIR. The present work will engineer a tandem vector for electroporation of host cells for development of a permantently transfected cell line to enable commercial production of the new fusion protein. The fusion protein will be purified with 2 columns and validated with HIR binding assays, IDS enzyme activity, Western blotting, and other biochemical assays. The biological activity of the HIRMAb-IDS fusion protein will be verified with assays in MPS-II fibroblasts that measure fusion protein uptake, cell IDS enzyme activity, and clearance of glycosoaminoglycans. These phase I studies will enable future phase II studies that test the safety, efficacy, and pharmacokinetics in animals prior to human testing.

Public Health Relevance:
Mucopolysacchridosis Type II, or Hunter's Syndrome, patients develop severe brain abrormalities, owing to the genetic defect in the gene encoding the lysosomal enzyme, iduronate-2-sulphatase (IDS). Enzyme replacement therapy (ERT) is not effective for the brain, because the IDS do not cross the blood-brain barrier (BBB). This research will develop a new treatment for the brain in patients with Hunter's Syndrome, which involves the re-engineering of human IDS as a fusion protein that crosses the human BBB.

Thesaurus Terms:
0-11 Years Old;4-Amino-10-Methylfolic Acid;4-Amino-4-Deoxy-10-Methylpteroyl-L-Glutamic Acid;5,6,7,8-Tetrahydrofolate[{..}]nadp+ Oxidoreductase;Affect;Affinity;Affinity Chromatography;Animals;Assay;Binding;Binding (Molecular Function);Bioassay;Biochemical;Biologic Assays;Biological;Biological Assay;Blood - Brain Barrier Anatomy;Blood-Brain Barrier;Blotting, Western;Body Tissues;Brain;Brain Diseases;Brain Disorders;Cho Cells;Cos Cells;Cos-1;Cell Line;Cell Lines, Strains;Cell Membrane;Cellline;Cells;Chemicals;Child;Child Youth;Children (0-21);Chimera Protein;Chimeric Proteins;Chinese Hamster;Chinese Hamster Ovary Cell;Chromatography, Affinity;Chromatography, High Performance Liquid;Chromatography, High Pressure Liquid;Chromatography, High Speed Liquid;Cloning;Culture Media, Serum-Free;Cytoplasmic Membrane;Dhfr;Dna;Dna Sequence;Deoxyribonucleic Acid;Deterioration;Development;Dihydrofolate Dehydrogenase;Dihydrofolate Reductase;Disease;Disorder;Dose;Drug Kinetics;Drugs;Ec 1.5.1.3;Elisa;Electroporation;Encephalon;Encephalon Diseases;Encephalons;Engineering;Engineerings;Enzyme-Linked Immunosorbent Assay;Enzymes;Equipment;Evaluation;Exclusion;Exhibits;Fibroblasts;Folic Acid Reductase;Fusion Protein;Future;Gamma Globulin, 7s;Gargoylism, Hunter Syndrome;Gene Amplification;Genes;Genetic Alteration;Genetic Change;Genetic Condition;Genetic Diseases;Genetic Defect;Genital System, Female, Ovary;Hplc;Hemato-Encephalic Barrier;Hereditary;Hereditary Disease;High Pressure Liquid Chromatography;Hour;Human;Human Engineering;Human, Child;Human, General;Hunter Corrective Factor;Hunter Syndrome Gargoylisms;Hunter Syndrome;Hunter's Syndrome;Hunter-Fraser Syndrome;Hunter-Mcalpine Syndrome;Hydrocephalus;Hydrocephaly;Hypoxanthines;Insr;Insr Protein, Human;Iduronate Sulfatase;Iduronate Sulfate Sulfatase;Iduronatesulfate Sulfohydrolase;Igg;Immunoglobulin G;Inherited;Insulin Receptor;Insulin Receptor Protein-Tyrosine Kinase;Insulin-Dependent Tyrosine Protein Kinase;Intracranial Cns Disorders;Intracranial Central Nervous System Disorders;L-Glutamic Acid, N-(4-(((2,4-Diamino-6-Pteridinyl)Methyl)Methylamino)Benzoyl)-;L-Iduronate-2-Sulfate 2-Sulfohydrolase;Label;Light;Mps Ii;Man (Taxonomy);Man, Modern;Measures;Medication;Mental Retardation;Methotrexate;Methotrexate Methylaminopterin;Methotrexatum;Metotrexato;Moab, Clinical Treatment;Molecular Disease;Molecular Interaction;Monoclonal Antibodies;Mucopolysaccharidosis 2;Mucopolysaccharidosis Ii;Mutation;Nerve Cells;Nerve Unit;Nervous System, Brain;Neural Cell;Neurocyte;Neurons;Ovary;Patients;Peptide Receptor;Pharmaceutic Preparations;Pharmaceutical Preparations;Pharmacokinetics;Pharmacology And Toxicology;Phase;Photoradiation;Physiologic Pulse;Plasma Membrane;Production;Protein Replacement Therapy;Protein-Free Media;Proteins;Publishing;Pulse;Recombinant Fusion Proteins;Recombinants;Research;Robotics;Sbir;Sbirs (R43/44);Sds Page;Sds Polyacrylamide Gel Electrophoresis;Scientist;Secondary To;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;Structure;Sulfoiduronate Sulfatase;Testing;Tetrahydrofolate Dehydrogenase;Therapeutic;Thymidin;Thymidine;Time;Tissues;Transgenes;Triage;Western Blotting;Western Blottings;Western Immunoblotting;Work;X-Linked Hurler Syndrome;Affinity Chromatography;Affinity Purification;Blood Brain Barrier;Brain Cell;Children;Cultured Cell Line;Disease /Disorder;Disease/Disorder;Drug /Agent;Drug Development;Drug/Agent;Enzyme Activity;Enzyme Linked Immunosorbent Assay;Enzyme Replacement Therapy;Expression Vector;Gene Product;Genetic Disorder;Genome Mutation;Hereditary Disorder;High Performance Liquid Chromatography;Human Insr Protein;Iduronate Sulfatase (Ids) Deficiency;Iduronate Sulfatase Deficiency;Iduronate-2-Sulfatase;Iduronate-2-Sulfatase Deficiency;Insulin Receptor, Human;Molecular Trojan Horse;Molecular Trojan Horse Technology;Monoclonal Antibody;Mucopolysaccharide Storage Disease Ii;Mucopolysaccharidosis (Mps) Ii;Mucopolysaccharidosis Type Ii;Natural Gene Amplification;Neuronal;Peptidomimetics;Phase 1 Study;Phase 2 Study;Plasmalemma;Protein Blotting;Receptor Binding;Response;Safety Testing;Sulfo-Iduronate Sulfatase (Sids) Deficiency;Sulfo-Iduronate Sulfatase Deficiency;Uptake;Vector;Youngster

Type II Mucopolysaccharidosis (MPS), also known as Hunter's syndrome, is a genetic disease that affects the lysosomal enzyme, iduronate-2-sulfatase (IDS). Consequently, patients with MPS-II develop extensive lysosomal storage product accumulation in tissues, including the brain. Children born with MPS-II develop multiple brain disorders including mental retardation and hydrocephalus. The current therapy for MPS-II is Enzyme Replacement Therapy (ERT) with the recombinant human enzyme, IDS. However, ERT does not treat the brain of MPS-II, because IDS does not cross the blood-brain barrier (BBB). The present work will continue work on the development of a new treatment of the brain of MPS-II, which is a genetically engineered biopharmaceutical fusion protein. The IDS is genetically fused to a BBB molecular Trojan horse, which is a genetically engineered peptidomimetic monoclonal antibody (MAb) against an endogenous BBB peptide receptor, the human insulin receptor (HIR). The human IDS is fused to the heavy chain of the HIRMAb to create a new chemical entity, called the HIRMAb-IDS fusion protein. Phase I studies show the HIRMAb-IDS fusion protein could be engineered and expressed by stably transfected host cells. The fusion protein retained high affinity binding to the HIR and retained high IDS enzyme activity. The proposed phase II work will develop a manufacturing plan that can be replicated for future GMP manufacturing. The AGT-182 produced in phase II will be evaluated for safety, toxicology and pharmacokinetics in 8 Rhesus monkeys. The completion of this work will enable entry of the AGT-182 drug development program into GLP toxicology and GMP manufacturing required for submission of an IND to begin treatment of the brain in patients with MPS-II.

Public Health Relevance:
Mucopolysacchridosis Type II, or Hunter's Syndrome, patients develop severe brain abrormalities, owing to the genetic defect in the gene encoding the lysosomal enzyme, iduronate-2-sulphatase (IDS). Enzyme replacement therapy (ERT) is not effective for the brain, because the IDS does not cross the blood-brain barrier (BBB). This research will develop a new treatment for the brain in patients with Hunter's Syndrome, which involves the re-engineering of human IDS as a fusion protein that crosses the human BBB.

Public Health Relevance Statement:
Mucopolysacchridosis Type II, or Hunter's Syndrome, patients develop severe brain abrormalities, owing to the genetic defect in the gene encoding the lysosomal enzyme, iduronate-2-sulphatase (IDS). Enzyme replacement therapy (ERT) is not effective for the brain, because the IDS does not cross the blood-brain barrier (BBB). This research will develop a new treatment for the brain in patients with Hunter's Syndrome, which involves the re-engineering of human IDS as a fusion protein that crosses the human BBB.

Project Terms:
Adult; Affect; Affinity; Affinity Chromatography; Anions; Antibodies; Binding (Molecular Function); Biochemical; Biological Products; Bioreactors; Blood; Blood - brain barrier anatomy; Brain; brain cell; Brain Diseases; Cations; Cells; Cerebrospinal Fluid; Chemicals; Child; Chimeric Proteins; Chinese Hamster; Chinese Hamster Ovary Cell; Chromatography; Clinical Trials; design; Development; Disease; Dose; Drug Delivery Systems; drug development; Drug Kinetics; Engineering; enzyme activity; enzyme replacement therapy; Enzyme-Linked Immunosorbent Assay; Enzymes; Exhibits; Filtration; Future; Genes; Growth; Hereditary Disease; Human; Human Engineering; human INSR protein; Hydrocephalus; iduronate-2-sulfatase; Immunoglobulin G; Insulin Receptor; Macaca mulatta; Measurement; Measures; Mediating; Membrane; Mental Retardation; molecular trojan horse; Monoclonal Antibodies; Mucopolysaccharidosis II; Mutation; nano; Neurons; neuropathology; Organ; Ovary; Patients; pediatrician; Penetration; Peptide Receptor; peptidomimetics; Perfusion; Peripheral; Pharmaceutical Preparations; Phase; phase 1 study; Plasma; Primates; Process; Production; programs; Progress Reports; Protein Binding; Proteins; receptor; Recombinant Fusion Proteins; Recombinants; Research; Safety; Serum-Free Culture Media; Small Business Innovation Research Grant; Structure; Temperature; Testing; Tissues; Toxicology; uptake; Validation; Work