SBIR-STTR Award

We previously demonstrated in mice that the major pathogenic features (eosinophilic airway inflammation and airway hyperresponsiveness [AHR]) were markedly enhanced in mice exposed to repeated social stress and these changes were associated with impaired glucocorticoid receptor (GR) expression and function. In studies on non-human primates (rhesus macaques) at the California National Primate Research Center (CNPRC) at UC Davis, anxious temperament (an early manifestation of psychosocial stress) in young monkeys was significantly associated with AHR and glucocorticoid non-responsiveness of immune cells. We propose to investigate secoisolariciresinol diglucoside (SDG), a novel antiinflammatory molecule, and natural non-toxic bioactive component of flaxseed, as an alternative/adjunct therapy for glucocorticoid resistant asthma. SDG has proven reducing, chelating and free radical scavenging activities in vitro and in vivo and it induces activation of nuclear factor erythroid 2-related factor 2 (NRF2) a major anti-oxidant transcription regulator and inhibitor of NF-kB. Our aim 1 is to evaluate the effects of the synthetic version of SDG (LGM2605) on airway hyperresponsiveness (AHR), inflammatory markers of blood and bronchoalveolar lavage (BAL) immune cells and glucocorticoid function of peripheral blood mononuclear cells of socially stressed rhesus macaques. Aim 2 is to assess LGM2605 treatment on genetic and epigenetic regulation of the GR, NF-kB and NRF2 using bronchial brush biopsies. The strength and novelty of our proposal is the unique Rhesus macaque model system of asthma associated with psychosocial stress and glucocorticoid resistance, and the synthetic availability of LGM2605 through LignaMed. Upon completion, these studies will lay the groundwork for subsequent Phase II trial and ultimately to human clinical projects.

Public Health Relevance Statement:
NARRATIVE: We previously found in murine, non-human primate and patient studies that severe asthma was associated with psychosocial stress and glucocorticoid resistance. Here we will establish the effects of the synthetic version of SDG (LGM2605), a natural flaxseed lignan compound with demonstrated strong antioxidant properties, on a stress-related asthma model in non-human primates. Our studies will guide a Phase II project to verify the effectiveness and mechanism of action of this compound leading to investigations in asthma patients and ultimately to an alternative/adjunct therapy of glucocorticoid resistant asthma.

Project Terms:
Aftercare; airway hyperresponsiveness; airway inflammation; Allergic; Animals; Anti-Inflammatory Agents; Antioxidants; Anxiety; anxious temperament; Area; Asthma; asthmatic patient; Biological; Biological Models; Biopsy; Biopsy Specimen; Blood; Brain; Bronchoalveolar Lavage; California; Cells; Chelating Agents; Child; Chronic; Clinical; cohort; Disease; DNA Methylation; Effectiveness; Epigenetic Process; epigenetic regulation; Exhibits; Flax; flaxseed lignan; Free Radical Scavenging; Genes; Genetic; Genetic Transcription; Global Change; Glucocorticoid Receptor; Glucocorticoids; High Pressure Liquid Chromatography; Histone H3; Human; Immune; Impairment; in vitro activity; in vivo; Inflammatory; inflammatory marker; inhibitor/antagonist; Investigation; Lead; Lysine; Macaca mulatta; methylation pattern; Modeling; Monkeys; Mus; Natural Products; NF-kappa B; nonhuman primate; novel; NR3C1 gene; nuclear factor-erythroid 2; Pathogenicity; Pathway interactions; Patients; peripheral blood; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; phase II trial; Phylogenetic Analysis; Play; Predisposition; Prevalence; Price; Primates; Production; promoter; Property; psychosocial; Psychosocial Stress; receptor expression; receptor function; Receptor Gene; Research; Resistance; Rodent; Role; scale up; secoisolariciresinol; social stress; Stress; Testing; transcription factor; Work

IND enabling development of LGM2605 as adjuvant treatment for asthma Glucocorticoid resistance is a major treatment problem in asthma. Our recent studies in mice, non-human primates and severe asthma patients, along with reports by others suggest that glucocorticoid receptor (GR) expression was impaired by psychosocial stress, in association with enhanced NF-kB activation and glucocorticoid non-responsiveness of immune cells. This Phase II STTR proposal was developed jointly between LignaMed (Dr. Sielecki), UPenn (Dr. Christofidou-Solomidou and UC Davis (Dr. Haczku) based on our results generated by the Phase I STTR project. Our study strongly suggested that LGM2605, a racemic synthetic form of a novel, natural, non-toxic, anti-inflammatory component of flaxseed, secoisolariciresinol diglucoside (SDG) may be effective to treat severe asthma exacerbation induced by inhalation of ozone. LGM2605 is a racemate which has proven free radical scavenging activities in vitro and in vivo and it induces activation of nuclear factor erythroid 2-related factor 2 (NRF2) a major anti-oxidant transcription regulator and inhibitor of NF-kB. As part of assembling an IND package we will complete a pivotal proof of concept study using rhesus macaques, because of their phylogenetic proximity to humans with a high degree of immune crossreactivity and a predisposition to spontaneously develop both asthma and psychosocial stress. Our exciting preliminary results from stressed asthmatic macaques treated with LignaMed’s LGM2605 demonstrated a significant suppression of airway inflammation and it abolished airway hyperresponsiveness (AHR) in response to ozone exposure. We hypothesize that LGM2605 alleviates asthma symptoms by interfering with activation of immune and airway structural (epithelial and smooth muscle) cells and improving glucocorticoid responsiveness through activation of NRF2 gene expression and downstream anti-oxidant pathways. Aim 1. Assess the mechanism of action and dose-dependent effects of LGM2605 treatment on regulation of the GR, NF-kB and NRF2 expression and glucocorticoid responsiveness in vitro. Aim 2. A. Scale up and definition of release specifications of the single isomer of LGM2605; B. Pharmacokinetic evaluation of the single isomer of LGM2605 in non- human primates (rhesus macaques). Aim 3. Study the dose-dependent effects of single isomer LGM2605 on preventing AHR, immune cell activation and improving glucocorticoid responsiveness in ozone- exposed rhesus macaques. Our translational approach using rhesus macaques (in vivo clinical testing) and cells from severe asthma patients (in vitro mechanistic studies on cell types relevant to asthma) will establish how LGM2605 affects glucocorticoid responsiveness and will lay the groundwork for subsequent human clinical trials.

Public Health Relevance Statement:
NARRATIVE: In collaboration with UC Davis and the University of Pennsylvania LignaMed LLC is developing LGM2605 as an adjuvant treatment of severe asthma, a condition associated with heightened airway inflammation and non- responsiveness to mainstream corticosteroid treatment. Our Phase-I STTR study demonstrated effectiveness of LGM2605 in reducing airway hyperreactivity and airway inflammation in asthmatic rhesus macaques. In this Phase-II project we will use this macaque model for a pivotal proof of concept study. In addition, we will investigate the mechanism of action of this compound in peripheral blood mononuclear cells from severe asthma patients and in human airway smooth muscle cells by hypothesizing that LGM2605 inhibits NF-kB and improves glucocorticoid responsiveness through stimulating NRF2-related pathways. These studies will determine the optimal dosing regimen, establish the role of the oxidative molecular changes in glucocorticoid responsiveness and will inform future clinical trials of LGM2605 as an asthma treatment.

NIH Spending Category:
Asthma; Climate-Related Exposures and Conditions; Genetics; Lung; Prevention

Project Terms:
Adjuvant; Adrenal Cortex Hormones; Affect; Aftercare; Age; airway hyperresponsiveness; airway inflammation; airway obstruction; Animals; Anti-inflammatory; Antioxidants; Asthma; asthma exacerbation; asthmatic; asthmatic patient; Automobile Driving; base; biobank; Biological; Biological Assay; Biological Availability; Blood; Bronchoalveolar Lavage; Categories; Cell Count; Cell Culture Techniques; Cell Line; cell type; Cells; Clinic; Clinical Trials; cohort; Collaborations; Dendritic Cells; design; Development; Dexamethasone; Dose; Drug Kinetics; Effectiveness; Enrollment; enterodiol; Epigenetic Process; Epithelial; Epithelial Cells; Evaluation; Flax; Free Radical Scavenging; Future; Gender; Gene Expression; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; healthy volunteer; Human; Immune; immune activation; Immune Cell Activation; Impairment; improved; In Vitro; in vitro activity; in vivo; Inflammation; inflammatory marker; Inhalation; inhibitor/antagonist; Isomerism; Macaca; Macaca mulatta; Mainstreaming; methacholine; Modeling; Molecular; Mus; Myelogenous; NF-kappa B; nonhuman primate; novel; NR3C1 gene; nuclear factor-erythroid 2; Optics; Oral; Ozone; ozone exposure; Pathway interactions; Pennsylvania; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Phylogenetic Analysis; Predisposition; prevent; Psychosocial Stress; Pulmonary Surfactant-Associated Protein D; receptor expression; receptor function; Receptor Gene; Regimen; Regulation; Reporting; research clinical testing; Resistance; respiratory smooth muscle; response; Role; Sampling; scale up; secoisolariciresinol; Small Business Technology Transfer Research; Smooth Muscle Myocytes; Steroid Resistance; Stress; Structure; Symptoms; Testing; transcription factor; transcriptome sequencing; translational approach; Universities