SBIR-STTR Award

This proposal brings together a top team with diverse and complementary expertise to develop a novel therapeutic agent based on a newly described serum protein, renalase (RNLS) to treat acute pancreatitis (AP). This protein appears to have a novel protective effect in models of acute tissue injury, including AP. AP affects more than 250,000 people/year in the USA and can cause death in 30% of those with severe disease. It is the most common reason for hospitalization in the USA for individuals with gastrointestinal disease. RNLS is produced in the kidneys and other tissues including the pancreas. Initial data show that: 1) Serum RNLS rapidly decreases soon after the onset of AP in a murine model and in humans; 2) Genetic deletion of RNLS increases the severity of experimental acute murine pancreatitis; 3) Recombinant RNLS (rRNLS) reduces pancreatitis responses in isolated murine pancreatic acinar cells; 4) Exogenous rRNLS significantly reduces the severity of acute murine pancreatitis in vivo when given either as pretreatment or two hours after the onset of disease. Initial studies show that RNLS-derived peptides reduce acute injury in several types of cultured cells. This protective effect has been localized to the C-terminal region of RNLS. These findings suggest that RNLS will have a protective role in AP. The proposed studies will examine RNLS-derived peptides and compare their potency to rRNLS in reducing AP injury. The peptides may have advantages over rRNLS by way of ease of manufacture and stronger patent position. Lead drugs will be selected and examined with the following Specific Aims: 1) RNLS and RNLS-derived, smaller peptides will be compared/prioritized for their potency in reducing pancreatitis responses in isolated murine pancreatic acinar cells. 2) The effects on AP severity of rRNLS and the two most potent peptides identified in Aim 1 will be examined in two experimental murine models: a mild form of AP induced by high doses of cerulein and a severe model of AP induced by intraperitoneal alcohol and a fatty acid. To further reflect clinical treatment scenarios, the effectiveness of rRNLS or related peptides will be examined when given prior to the onset of injury or after the onset of disease. Further, traditional biochemical measures of AP will be correlated with RNLS levels as well as pain, a key symptom of acute pancreatitis. With success the project will be poised for late stage development and clinical studies of a novel and rationally designed therapeutic. Based on the strength of the preliminary data and evidence that this novel survival pathway is fundamentally important in both murine models and human disease, RNLS therapy has a very high potential for being effective in preventing or treating AP.

Public Health Relevance Statement:
Project Narrative Acute pancreatitis (AP) is the most frequent gastrointestinal cause for hospital admission in the US, exacting a severe toll on patients and the health care system. We propose to study whether renalase (RNLS), a naturally occurring circulating growth factor whose deficiency is linked to AP, or a RNLS peptide agonist, is a therapeutically equal or better drug candidate. This project combines the efforts of Drs. Gary Desir, who pioneered the discovery and biology of RNLS, and Fred Gorelick, an expert in the mechanisms and clinical features of AP, with those of an experienced drug development team of Bessor Pharma. It builds on promising initial results showing preclinical proof of concept that the RNLS agonist approach is effective in vitro and in vivo.

Project Terms:
abstracting; Acinar Cell; Acute; acute pancreatitis; Admission activity; Affect; Agonist; Alcohols; Amylases; base; Binding; Biochemical; Biology; body system; C-terminal; Ca(2+)-Transporting ATPase; Calcium; Cause of Death; Cell Death; Cell membrane; Cell model; Cell Survival; Cells; Cisplatin; Clinical; Clinical Research; Clinical Treatment; Creatinine; Cultured Cells; Data; design; Development; Disease; Dose; drug candidate; drug development; Edema; Effectiveness; Enzyme Precursors; Ethanol; experience; Fatty Acids; gastrointestinal; Gastrointestinal Diseases; Generations; Genetic; Growth; Healthcare Systems; Hospitalization; Hospitals; Hour; Human; human disease; IL6 gene; In Vitro; in vivo; Incidence; Individual; Infiltration; Inflammation; Inflammation Mediators; Inflammatory Response; Injury; Innovative Therapy; intraperitoneal; Kidney; Lead; Legal patent; Link; Lung; Lung Inflammation; macrophage; Measures; Mediating; Metabolism; mimetics; Modeling; mouse model; Mus; neutrophil; novel; novel therapeutics; Onset of illness; Pain; palmitoleic acid; Pancreas; Pancreatitis; Pathologic; Pathway interactions; Patients; peptide analog; Peptides; Peroxidases; Pharmaceutical Preparations; Positioning Attribute; pre-clinical; prevent; protective effect; Protein Engineering; Proteins; Pulmonary Edema; Recombinants; response; Role; Serum; Serum Proteins; Severities; Severity of illness; Signal Transduction; Staging; success; Symptoms; Testing; Therapeutic; Therapeutic Agents; Time; Tissue Survival; Tissues; Trypsin; Trypsinogen

This proposal brings together a top team with diverse and complementary expertise to develop a novel therapeutic agent based on a newly described serum protein, renalase (RNLS) to treat acute pancreatitis (AP). This protein has a novel protective effect in models of acute tissue injury, including AP. AP affects more than 250,000 people/year in the USA and can cause death in 20-30% of those with severe disease. It is the most common reason for hospitalization in the USA for individuals with gastrointestinal disease. RNLS is produced in the kidneys and other tissues, including the pancreas. Initial data show that: 1) Serum RNLS rapidly decreases soon after the onset of AP in a murine model and in humans, thus suggesting that RNLS has biomarker potential; 2) Genetic deletion of RNLS increases the severity of experimental acute murine pancreatitis; 3) Human recombinant research grade RNLS (hrRNLS) reduces pancreatitis responses in isolated murine pancreatic acinar cells; 4) Exogenous hrRNLS significantly reduces the severity of acute murine pancreatitis in vivo when given either as pretreatment or two hours after the onset of disease. The full-length hrRNLS protein was more efficacious in in vitro and in vivo AP models than hrRNLS peptides. These findings suggest that hrRNLS will have a protective role in AP. The proposed studies will examine research grade hrRNLS without an affinity tag present in previous drug supply synthesized in E. coli. We will test hrRNLS in mild (cerulein) and severe disease (arginine) AP models. The studies will determine the concentration-dependent effects and the relative effects of route (subcutaneous and IV) to reduce key measures of AP. hrRNLS will be given prior to the onset of injury (prophylactic) or after onset (therapeutic) of AP. In addition to quantifying biochemical and histologic parameters of AP in the pancreas and blood, the kidney and lung (also damaged in AP) will be examined for injury in male and female mice. Since severe pain is a major sequela of AP, hrRNLS effects to reduce sensitization to thermal and mechanical stimuli will be assessed. Pharmacokinetics of both routes will be examined. Initial toxicity studies of hrRNLS will be performed to determine its therapeutic index. Lastly, hrRNLS will be produced under GMP conditions and used in additional pharmacological studies towards an IND. Success in this project would advance hrRNLS as the potential 1st to market drug to treat AP.

Public Health Relevance Statement:
PROJECT NARRATIVE Acute pancreatitis (AP) is the most frequent gastrointestinal cause of hospital admission in the U.S., exacting a severe toll on patients and the health care system. The research supported by this grant will advance the development of recombinant human renalase for the treatment of this severe condition for which there is not approved drug treatment in the US. Renalase is a naturally occurring circulating pro-survival cytokine and immuno-inflammatory regulator; its deficiency is linked to AP and its replacement may lessen injury and improve survival.

Project Terms:
Absence of pain sensation; Acinar Cell; Acute; acute pancreatitis; acute toxicity; Advanced Development; Affect; Affinity; Amylases; Arginine; base; Biochemical; Biological; Biological Assay; Biological Markers; Blood; Calcium; Cause of Death; cell injury; Cell Line; Cellular Assay; Chromatography; Chronic; Circular Dichroism; Clinical; Creatinine; Crystallization; cytokine; Data; Development; Development Plans; Disease; Disease model; Dose; drug candidate; drug development; Drug Kinetics; drug market; Drug Targeting; Edema; Endotoxins; Enzyme Precursors; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Evaluation; Female; Funding; gastrointestinal; Gastrointestinal Diseases; Gel Chromatography; Generations; Genetic; Government; Grant; Guidelines; Healthcare Systems; Histologic; Hospitalization; Hour; Human; human model; IL6 gene; improved; In Vitro; in vivo; in vivo Model; Individual; Inflammation; Inflammation Mediators; Inflammatory; inflammatory marker; Inflammatory Response; injured; Injury; innovation; Ion Exchange; Isoelectric Focusing; Kidney; Length; Link; Lung; macrophage; male; Maximum Tolerated Dose; Measures; Mechanics; Mediating; melting; Metabolism; Methods; Modeling; mortality; mouse model; Mus; Necrosis; neutrophil; novel; novel therapeutics; Onset of illness; Pain; Pancreas; Pancreatic Diseases; Pancreatitis; Pathologic; Patients; peptide analog; Peptides; Peroxidases; Persons; Pharmaceutical Preparations; Pharmacology Study; Pharmacotherapy; Phase; Plasma; prevent; Production; prophylactic; protective effect; Proteins; Proto-Oncogene Proteins c-akt; Protocols documentation; Quality Control; receptor binding; Recombinant Proteins; Recombinants; renal damage; Research; Research Support; response; Role; Route; Serum; Serum Proteins; Severities; Signal Transduction; Small Business Innovation Research Grant; STAT3 gene; Stimulus; Structure; subcutaneous; success; Testing; Therapeutic; Therapeutic Agents; Therapeutic Index; Tissues; Toxic effect; Trypsin; Trypsinogen; Western Blotting