SBIR-STTR Award

Advent Therapeutics Inc. (Advent) is a biotech company focusing on the development, reformulation and optimized delivery of legacy drugs to address serious unmet medical needs in underserved patient populations. Advent is developing an aerosol formulation of its proprietary optimized, water miscible vitamin A (vitA) palmitate for non-invasive (inhaled) delivery of the drug to preterm infants. Our innovative inhaled vitA formulation 1) avoids the drawbacks of invasive intramuscular (IM) injections, the current dosing modality, and 2) provides both direct-to-target-organ delivery to prevent bronchopulmonary dysplasia (BPD), known also as chronic lung disease (CLD) of prematurity, and systemic delivery for treating vitA deficiency (VAD) in premature neonates. Our preliminary in vitro testing shows that our vitA formulation can be efficiently aerosolized by a commercial nebulizer (Aerogen Aeroneb Pro®), producing optimally sized particles for efficient inhalation dosing. Early in vivo studies with our collaborator, Dr. Virender Rehan at Harbor-UCLA Medical Center, in a pre-weaned rat model, show that inhaled vitA leads to a rise in serum vitA levels, indicating systemic delivery. Additionally, in the lung, levels of protein and mRNA of genes involved in lung maturation are upregulated, providing direct evidence that inhaled vitA stimulates the developing lung. All metrics showed significant improvements in these early studies. Our proposal describes an expanded collaboration, with in vivo work in Dr. Rehan?s lab and shared efforts on assays/analysis. The consequences of VAD including BPD, retinopathy of prematurity, and sepsis are all costly complications of prematurity, with significant risk of poor clinical outcomes. Currently available vitA therapies either require invasive IM injection, or are generally inadequate for premature neonates (enteral or IV routes). Inhalation dosing overcomes all of these hurdles, enabling higher vitA utilization, reducing risk and cost for BPD/CLD. Our goals in Phase I are to demonstrate in a well characterized pre-weaned rat model that: 1) our vitA formulation delivered by non-invasive inhalation is at least as effective as invasive IM injection, and 2) that inhaled vitA can attenuate hyperoxic lung injury, thereby reducing the risk for development of BPD. To achieve this, our specific Aim 1 will measure the effectiveness of inhalation of our proprietary vitA formulation, aerosolized by the Aeroneb Pro®, on raising serum vitA and stimulating lung maturation in pre-weaned rats, comparing to effects of IM vitA dosing as a baseline. Analysis of lung and liver tissue will be by Western blot, RT-PCR, and immunohistochemistry. Vitamin A and retinol binding protein will by assayed by using ELISA. Our measure of success will be direct demonstration of at least similar effects with aerosolized vitA vs. typical IM dosing (in comparison to untreated control groups). In specific Aim 2, we will evaluate the effect of inhaled vitA on attenuating hyperoxia-induced neonatal lung injury as compared to IM delivery. Our measure of success will be efficacy in reducing hyperoxic lung damage equal to or better than IM dosing and relevant controls.

Project Terms:
Address; adipocyte differentiation; aerosolized; Aerosols; All-Trans-Retinol; Alveolar; Animal Model; Applications Grants; Attenuated; base; Benchmarking; Biological Assay; Biological Markers; Biological Models; Biotechnology; Blood Circulation; Bronchopulmonary Dysplasia; CCAAT-Enhancer-Binding Proteins; Chronic lung disease; Clinical; Collaborations; Communities; compare effectiveness; Confidential Information; Control Groups; cost; Data; design; Development; Diagnosis; Dose; Drug Delivery Systems; drug development; Effectiveness; effectiveness measure; Enteral; Enzyme-Linked Immunosorbent Assay; FDA approved; Formulation; Future; Gene Expression; Genes; Goals; Hyperoxia; Immunohistochemistry; In Vitro; in vitro testing; in vivo; Inflammation; Inhalation; Injectable; Injections; innovation; interest; Intramuscular; Intramuscular Injections; Letters; Liver; Lung; Lung diseases; lung injury; lung maturation; Measures; Medical; Medical center; Messenger RNA; Modality; Modeling; Molecular; Morphology; National Heart, Lung, and Blood Institute; Nebulizer; Neonatal; neonatal lung injury; neonatal sepsis; novel; novel strategies; Organ; Orphan; Outcome; Oxygen; particle; Particle Size; Pathway interactions; patient population; Patients; Peroxisome Proliferator-Activated Receptors; Pharmaceutical Preparations; Phase; Phospholipids; Physiology; Pilot Projects; Population; premature; Premature Infant; premature neonates; prevent; Prevention; programs; Proteins; Pulmonary Surfactant-Associated Protein B; Pulse Oximetry; Rattus; receptor; Regimen; Research; Respiratory physiology; response; Retinol Binding Proteins; Retinopathy of Prematurity; retinyl palmitate; Reverse Transcriptase Polymerase Chain Reaction; Risk; Route; RXR; Sepsis; Serum; Small Business Innovation Research Grant; standard of care; Structure of parenchyma of lung; success; Supplementation; surfactant; Therapeutic; Time; Tissues; Toxic effect; Vitamin A; Vitamin A Deficiency; Water; Weaning; Western Blotting; Work;

Advent Therapeutics Inc. (Advent) is a biotech company focusing on the development, reformulation and optimized delivery of legacy drugs to address serious unmet medical needs in the underserved neonatal and pediatric patient populations. Advent is developing an aerosol formulation of its proprietary, optimized water miscible vitamin A (vitA) palmitate for non-invasive (inhaled) delivery to preterm infants to address vitA deficiency (VAD) and associated serious complications such as bronchopulmonary dysplasia (BPD, the focus of our Phase I SBIR and this Phase II application), retinopathy of prematurity (ROP), and neonatal sepsis – all costly complications with significant morbidity/mortality. Our innovative inhaled (non-invasively dosed) vitA formulation 1) avoids the drawbacks of invasive intramuscular (IM) injections and absorption limitations of current oral forms, overcoming significant hurdles to more frequent NICU utilization, and 2) provides direct-to-target-organ delivery for increased efficacy, with our Phase I in vivo data showing significant benefit over IM dosing in mitigating hyperoxic lung damage (our BPD animal model), while providing adequate systemic delivery to also treat VAD. In collaboration with Dr. Virender Rehan at Harbor-UCLA Medical Center, we have accomplished our Phase I Specific Aims, demonstrating that: 1) inhaled vitA stimulates lung maturation as demonstrated via assay of lung biomarkers showing upregulation of retinol receptors, surfactant protein and phospholipid synthesis, and maturation biomarkers while simultaneously raising serum vitA levels similar to IM dosing; and 2) inhaled vitA dramatically (vs IM) reduces hyperoxic lung tissue damage via examination of lung tissue histomorphometry and reduction of lung-injury biomarkers. In Phase II, we will further refine the inhaled vitA dosing strategy for mitigating hyperoxic lung damage in a step-wise approach by studying the well characterized pre-weaned rat model as in Phase I and then expanding our studies to a pre-term rabbit model, with lung maturation status more closely mimicking human preterm infant lung to allow for translation of our findings into the clinical. Phase II Specific Aims are: 1: Optimize the dosing regimen of aerosolized vitA for mitigating hyperoxic lung damage in our rat model for the “neonatal” timeframe (acute phase) and long term sequalae into adulthood (chronic phase) using similar biomarkers and morphologic evaluation as per Phase I. Aim 2: Extend acute and chronic phase benefits of inhaled vitA to the premature rabbit model. Aim 1 & 2 measures of success will be demonstration of improved lung maturation and mitigation of lung injury vs IM-dosed controls, with an ideal outcome of showing lung status similar to healthy normal controls. Aim 3: Optimize aerosol characterization/delivery (initial in vitro experiments done concurrent with Aim 1), and subsequently conduct in vivo IND-enabling toxicology/PK studies. Aim 3 measures of success will be generation of data supporting further development of a potentially superior, non-invasive therapy for preventing BPD (and treating VAD), which will have significant clinical, financial, and societal implications.

Public Health Relevance Statement:
The consequences of inadequately treated vitamin A deficiency (VAD) in premature neonates including increased risk for bronchopulmonary dysplasia (BPD), retinopathy of prematurity (ROP) and sepsis, are associated with poor clinical outcomes in these fragile patients and require costly long-term treatment. Advent Therapeutics is developing an aerosol formulation of its proprietary optimized, water miscible vitamin A palmitate (vitA) for non-invasive delivery of the drug to preterm infants, enabling dosing by inhalation and thus obviating the need for intramuscular (IM) injection, and allowing potential increased effectiveness by delivery directly to the lung (a direct-to-target-organ treatment for BPD), while simultaneously providing systemic vitamin A availability important for treating VAD and the other related complications. Dosing vitA by inhalation should overcome current hurdles limiting vitamin A supplementation use in the NICU, enabling much higher vitamin A utilization, thereby reducing risk and cost of BPD in preterm infants.

Project Terms:
absorption; Acute; Address; Adult; aerosolized; Aerosols; All-Trans-Retinol; Animal Model; Biochemistry; Biological Assay; Biological Markers; Biotechnology; Bronchopulmonary Dysplasia; Chronic lung disease; Chronic Phase; Clinical; clinical application; Clinical Trials; Collaborations; Confidential Information; cost; Data; design; Development; Dose; Drug Delivery Systems; drug development; Effectiveness; Enteral; Evaluation; experimental study; FDA approved; first-in-human; Formulation; Funding; Future; Generations; Goals; Human; Hyperoxia; improved; In Vitro; in vivo; Industry Standard; Inhalation; Injections; innovation; interest; Intramuscular; Intramuscular Injections; Lung; Lung diseases; lung injury; lung maturation; Measures; Medical; Medical center; Metabolic; Modeling; Morbidity - disease rate; Morphology; mortality; National Heart, Lung, and Blood Institute; Neonatal; Neonatal Hyperoxic Injury; neonatal patient; neonatal sepsis; novel strategies; Oral; Organ; Orphan; Oryctolagus cuniculus; Outcome; Pathway interactions; patient population; Patients; pediatric patients; Peroxisome Proliferator-Activated Receptors; Pharmaceutical Economics; Pharmaceutical Preparations; Phase; Phospholipids; Population; Positioning Attribute; postnatal; premature; Premature Birth; Premature Infant; premature lungs; preterm newborn; prevent; Prevention; product development; Program Development; programs; Proteins; Pulmonary function tests; Pulmonary Surfactants; Rattus; receptor; Regimen; Respiratory physiology; Retinopathy of Prematurity; retinyl palmitate; Risk; Sepsis; Serum; Severities; Small Business Innovation Research Grant; specific biomarkers; Structure of parenchyma of lung; success; Sum; Supplementation; surfactant; Survival Rate; Testing; Therapeutic; Tissues; Toxicology; Translations; Up-Regulation; Vitamin A; Vitamin A Deficiency; Water; Weaning; Work