ArmaGen Technologies-Enabling Biotechnology for the Brain

Brain-penetrating biologic tumor necrosis factor inhibitor (TNFI) for Alzheimer's disease

AGT-110 is an IgG-decoy receptor fusion protein, and is the first brain-penetrating biologic tumor necrosis factor inhibitor (TNFI). The decoy receptor part is the human tumor necrosis factor-alpha receptor (TNFR) Type II extracellular domain, and the IgG part is a monoclonal antibody (MAb) against the human insulin receptor (HIR). The HIRMAb acts as a molecularTrojan horse to ferry the TNFR across the blood-brain barrier (BBB) via receptor-mediated transport on the endogenous BBB insulin receptor. Once behind the BBB, the fusion protein sequesters TNF-alpha, to block cytotoxic cytokine action in the central nervous system. AGT-110 can be developed for treatment of acute brain disease including brain trauma and spinal cord injury, and for chronic conditions, including neurodegeneration and depression.

AGT-110 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-110 is AGT-m110, which is a fusion protein of the human TNFR-II ECD and a chimeric MAb against the mouse transferrin receptor (TfR). The cTfRMAb-TNFR fusion protein (AGT-m110) has high brain uptake in the mouse, and is being tested in mouse models of neurodegeneration.

AGT-115 is an IgG-EPO fusion protein. Erythropoietin (EPO) is a potent neuroportective agent for acute brain disorders, such as stroke, traumatic brain injury, or spinal cord injury, and for chronic neurodegeneration, such as Parkinson's disease. However, EPO does not cross the blood-brain barrier (BBB). AGT-115 is a genetically engineered fusion protein comprised of a blood-brain barrier (BBB) molecular Trojan horse (MTH) protein that is fused to human EPO. AGT-115 binds simultaneously to 2 receptors. AGT-115 first binds to the human insulin receptor (HIR) on the BBB to initiate receptor-mediated transcytosis (RMT) across the BBB; this enables AGT-115 to enter brain from blood following intravenous administration. AGT-115 then binds the EPO receptor (EPOR) in the brain to trigger neuroprotection. AGT-115 has been genetically engineered to enable use in humans and binds with high affinity to both the human BBB HIR and the human neuronal EPOR.

The potential market for AGT-115 is large. There are nearly 800,000 strokes in the U.S. alone each year. About 70% of stroke patients survive the event with irreversible brain damage. The annual U.S. health care costs for rehabilitation of stroke patients is $45 billion. People suffering an acute stroke do not receive a neuroprotective agent, because no neuroprotective has been effective in stroke clinical trials. Neuroprotection is only possible in the first 3-4 hours after a stroke, and during this period the BBB is intact. The BBB problem is the single most important factor that has limited the development of neuroprotectives for stroke. The BBB is intact during the neuroprotection window, and any neuroprotectives developed for stroke must be able to cross the BBB.

AGT-115 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-115 is AGT-m115, which is a fusion protein of human EPO and a chimeric MAb against the mouse transferrin receptor (TfR). The cTfRMAb-EPO fusion protein (AGT-m115) has high brain uptake in the mouse, and is being tested in mouse models of stroke and neurodegeneration.

AGT-160

Brain-penetrating anti-amyloid antibody (AAA) for Alzheimer's disease

AGT-160 is a genetically engineered fusion protein where a singel chain Fv (ScFv) antibody against the Abeta amyloid peptide is fused to a genetically engineered HIRMAb. AGT-160 is the only anti-amyloid antibody (AAA) for Alzheimer's disease (AD) that penetrates the BBB in the absence of BBB disruption. AGT-160 is a fusion protein formed by 2 different proteins. The ScFv part of the molecule inhibits the formation of beta-amyloid plaque, which causes the dementia of AD. The other part of the fusion protein is a molecular Trojan horse that crosses the human blood-brain barrier via a specific receptor-mediated transport system. The present market for anti-amyloid drugs such as AGT-160 is >14 million in the U.S. alone, and includes the 6 million people suffering from AD as well as the 8 million people with mild cognitive impairment (MCI). Because the brain amyloid deposits up to 20 years before the onset of symptoms, the market for AGT-160 is even larger than the 14 million people with AD or MCI. The number of people with AD or MCI will increase to 20 million in the U.S. by 2020. it is important that anti-amyloid neurotherapeutics (that cross the blood-brain barrier) be started years before the onset of the clinical signs of AD. The delay in onset of the symptoms of AD by just 5 years reduces U.S. health care costs by $50 billion per year.

AGT-160 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-160 is AGT-m160, which is a fusion protein of the AAA and a chimeric MAb against the mouse transferrin receptor (TfR). The cTfRMAb-AAA fusion protein (AGT-m160) has high brain uptake in the mouse, and is being tested in mouse models of Alzheimer's disease.

Publications:

2007 Bioconjugate Chemistry

2010 Biotechnology and Bioengineering (primate study)

2010 Molecular Pharmaceutics (AGT-m160)

2011 Molecular Pharmaceutics (AGT-m160 AD mouse study)

AGT-181

Brain-penetrating iduronidase (IDUA) for Mucopolysaccharidosis (MPS) Type I (Hurler's syndrome)

AGT-181 is an IgG-enzyme fusion protein. AGT-181 is a new form of enzyme replacement therapy that is specifically indicated for the treatment of the brain for Mucopolysaccharidosis (MPS) Type I, or Hurler's syndrome. AGT-181 is a genetically engineered fusion protein. One part of the molecule is iduronidase (IDUA), the lysosomal enzyme that is missing in MPS-I that affects the brain. The other part of the molecule is the HIRMAb molecular Trojan horse that crosses the human blood-brain barrier via a specific receptor-mediated transport system. The bi-functional AGT-181 molecule is a novel biopharmaceutical that binds a specific human BBB receptor at the "head" of the molecule, and contains a lysosomal enzyme at the "tail" of the molecule. This lysosomal enzyme is missing in the brain, and other organs, of patients born with a specific lysosomal storage disorder.

There are over 40 Lysosomal Storage Disorders and about 75% of these severely affect the central nervous system (CNS). Conventional Enzyme Replacement Therapy (ERT) does not treat the brain in patients with Lysosomal Storage Disorders, because the enzyme therapeutic does not cross the BBB. Thus, the enzyme cannot penetrate into the brain from the blood. Enzyme replacement therapy of the a goal of ERT, and ArmaGen's solution to the BBB problem may enable enzyme replacement therapy of the brain with a conventional intravenous infusion, providing the missing enzyme is re-formulated to enable penetration of the BBB in humans.

AGT-181 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-181 is AGT-m181, which is a fusion protein of mouse IDUA and a chimeric MAb against the mouse transferrin receptor (TfR). The cTfRMAb-IDUA fusion protein (AGT-m181) produces a reversal of lysosomal inclusion bodies in the brain of the Hurler mouse following chronic intravenous bi-weekly injection.

Publications:

2008 Biotechnology and Bioengineering

2009 Journal of Biotechnology (primate toxicology study)

2011 Molecular Pharmaceutics (AGT-m181 Hurler mouse study)

AGT-182

Brain-penetrating iduronate-2-sulfatase (IDS) for MPS Type II (Hunter's syndrome)

AGT-182 is also an IgG-enzyme fusion protein. AGT-182 is a new form of enzyme replacement therapy that is specifically indicated for the treatment of the brain for Mucopolysaccharidosis (MPS) Type II, or Hunter's syndrome. AGT-182 is a genetically engineered IgG fusion protein. One part of the molecule is iduronate-2-sulfatase (IDS), the lysosomal enzyme that is missing in MPS-II that affects the brain. The other part of the molecule is the HIRMAb molecular Trojan horse that crosses the human blood-brain barrier via a specific receptor-mediated transport system. The bi-functional AGT-182 molecule is a novel biopharmaceutical that binds a specific human BBB receptor at the "head" of the molecule, and contains a lysosomal enzyme at the "tail" of the molecule. This lysosomal enzyme is missing in the brain, and other organs, of patients born with MPS-II.

AGT-182 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-182 is AGT-m182, which is a fusion protein of human IDS and a chimeric MAb against the mouse transferrin receptor (TfR).

Publications:

2010 Bioconjugate Chemistry

2011 Biotechnology & Bioengineering (primate study)

2011 Drug Metabolism & Disposition (AGT-m182)

AGT-185

Brain-penetrating paraoxonase (PON)-1 for chemical nerve gast treatment

AGT-185 is an IgG-paroxonase (PON)-1 fusion protein. Chemical nerve gas agents, such as organophosphates, cause morbidity and mortality via action within the central nervous system (CNS). Therefore, it is important that antidotes against chemical nerve gas agents are enabled to cross the human blood-brain barrier (BBB). The most potent human organophosphatase is PON-1, a serum protein. AGT-185 is a fusion protein of the HIRMAb human BBB molecular Trojan horse and human PON-1, with high hydrolytic activity against chemical nerve gas agents. AGT-185 is a bi-functional molecule that both crosses the human BBB and hydrolyzes organophosphates, and is the first organophosphatase enzyme ever engineered to specficially cross the human BBB. AGT-185 may also be therapeutic in cerebral atherosclerosis.

Publications:

2008 Molecular Pharmaceutics

2010 Biotechnology and Bioengineering (primate study)

AGT-190

Brain-penetrating glial derived neurotrophic factor (GDNF) for Parkinson's disease

AGT-190 is an IgG-GDNF fusion protein. Glial derived neurotrophic factor (GDNF) is a potent neuroportective agent for acute brain disorders, such as stroke, and for chronic neurodegeneration, such as Parkinson's disease. However, GDNF does not cross the blood-brain barrier (BBB). AGT-190 is a genetically engineered fusion protein comprised of the HIRMAb blood-brain barrier (BBB) molecular Trojan horse (MTH) protein that is fused to human GDNF. AGT-190 binds simultaneously to 2 receptors. AGT-190 first binds to the human insulinr eceptor (HIR) on the BBB to initiate receptor-mediated transcytosis (RMT) across the BBB; this enables AGT-190 to enter brain from blood following intravenous administration. AGT-190 then binds the GDNF receptor (GFR)-alpha-1 in the brain to trigger neuroprotection. AGT-190 has been genetically engineered to enable use in humans and binds with high affinity to both the human BBB HIR and the human neuronal GFRalpha-1.

AGT-190 is biologically active in humans and Old World primates such as the Rhesus monkey. The mouse-specific homologue of AGT-190 is AGT-m190, which is a fusion protein of human GDNF and a chimeric MAb against the mouse transferrin receptor (TfR). The cTfRMAb-GDNF fusion protein (AGT-m190) has high brain uptake in the mouse, and is therapeutic in a mouse model of Parkinson's disease.

Publications:

2008 Biotechnology and Bioengineering

2009 Drug Metabolism and Disposition (primate brain uptake study)

2009 Pharmaceutical Research (primate toxicology study)

2010 Drug Metabolism and Disposition (AGT-m190)

2010 Brain Research (AGT-m190 Parkinsons model)

2011 Drug Metabolism and Dispostion (AGT-m190 chronic dosing mouse model)

AGT-3

Brain-penetrating avidin-a universal brain delivery system for biotinylated therapeutics

AGT-3. AGT-3 is an IgG-avidin fusion protein. Avidin is fused to a genetically engineered monoclonal antibody (MAb) against the human insulin receptor (HIR). The avidin part binds any biotinylated therapeutic, including peptide radiopharmaceuticals for imaging brain diseases or short interfering RNA (siRNA). The HIRMAb part acts as a molecular Trojan horse to ferry the fused avidin across the blood-brain barrier (BBB) via transport on the endogenous HIR. Any biotinylated therapeutic, which normally does not cross the BBB, is able to penetrate the brain via transport mediated by attachment to the HIRMAb through the avidin-biotin linker. The AGT-3 fusion protein is formulated in one vial, and the mono-biotinylated drug is formulated in a second vial. The 2 vials are mixed just prior to intravenous administration. Owing to the extremely high affinity of avidin binding of biotin, there is instantaneous attachment of the biotinylated drug to the HIRMAb-avidin fusion protein. ArmaGen's avidin-based delivery system is encompassed within the broad claims of issued U.S. patent 6,287,792, which covers the use of avidin-biotin technology in drug delivery.

AGT-m1. The HIRMAb part of the AGT-3 fusion protein cross-reacts with the insulin receptor in Rhesus monkeys, but does not cross-react with the insulin receptor of rats or mice. There is no known monoclonal antibody (MAb) against the mouse insulin receptor that can be used as a BBB Trojan horse. Therefore, ArmaGen has engineered a surrogate Trojan horse that is active in the mouse, which is a chimeric MAb against the mouse transferrin receptor (TfR), and is designated the cTfRMAb or AGT-m1.

AGT-m3. A cTfRMAb-avidin fusion protein has been engineered, which is designated AGT-m3. The availability of AGT-m3 allows for preclinical testing in mouse models of the brain delivery of biotinylated therapeutics or diagnostics that are re-formulated with the Trojan horse-avidin fusion protein delivery system.

Publications:

2008 Bioconjugate Chemistry (AGT-3)

2009 Molecular Pharmaceutics (siRNA)

2009 Biotechnology and Bioengineering (AGT-m1)

2011 Bioconjugate Chemistry (AGT-m3)

AGT-3100

Alzheimer's disease amyloid brain scan: Brain-penetrating peptide radiopharmaceutical

AGT-3100 is a kit comprised of 2 vials. The first vial is the HIRMAb-avidin fusion protein (AGT-3). The second vial is biotinylated form of the Abeta (1-40) peptide of Alzheimer's disease (AD), which is modified to chelate heavy metal radionuclides. Abeta(1-40) is the most potent amyloid plaque imaging agent known, but cannot be developed as amyloid imaging agent, because the peptide does not cross the blood-brain barrier (BBB). The Abeta(1-40) peptide is re-formulated as AGT-3100. Upon mixing the 2 vials of the AGT-3100 kit, the biotinylated Abeta(1-40) peptide radiopharmaceutical is attached to the HIRMAb Trojan horse via a stable, high affinity avidin-biotin bond. Following intravenous injection, and owing to attachment to the HIRMAb Trojan horse, the Abeta(1-40) peptide is able to penetrate the BBB and to access amyloid plaques behind the BBB.

The mouse-specific homologue of AGT-3100 is designated AGT-m3100, which is also a kit comprised of 2 vials. The first vial contains the cTfRMAb-avidin fusion protein (AGT-m3). The second vial contains the N-biotinylated Abeta(1-40) peptide radiopharmaceutical. The AGT-m3100 penetrates the mouse brain rapidly, and 2.1% of injected dose (ID)/gram is taken up by the brain in the mouse in vivo. This level of brain uptake is comparable to the brain uptake of lipid soluble small molecule brain amyloid imaging agents. The small molecule brain amyloid imaging agents require the use of positron emission tomography (PET). AGT-3100 is radio-labeled with 111-indium for single photon emission computed tomography (SPECT).

2011 Bioconjugate Chemistry (AGT-m3100)

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