S250 Posters / Journal of Cystic Fibrosis 20S2 (2021) S1–S322
airway area (15.8 ± 0.42%). The methodology was validated by manually
quantifying EGFP-positive cells per total nuclei on a subset of airways, which
resulted in similar efficiencies (macro: 16.6 ± 2.8%, manual: 14.6 ± 2.4%)
Conclusion: We have designed several image analysis tools for ImageJ to
accelerate our pulmonary gene therapy research. Our workflow allows for
quantitative analysis of fluorescent reporter expression or RNAscope signal
in cell and tissue models for pulmonary gene therapy.
529
A phase 1b, randomized, double-blind, placebo-controlled, dose-
escalation trial of CB-280, an arginase inhibitor, in patients with
cystic fibrosis
S. Boas1, S. Donaldson2, K. McBennett3, T. Liou4, J. Howrylak5, L. Johnson6,
C. Teneback7, A. Dozor8, G. Sawicki9, J. Dumlao10, A. Pan10, L. Akella11,
J. Zhang12, S. Carroll10, K. Orford10, E. Kuriakose10, J. Mermis13. 1Cystic
Fibrosis Institute, Glenview, USA; 2Medicine, University of North Carolina at
Chapel Hill, Chapel Hill, USA; 3University Hospitals Cleveland Medical Center,
Cleveland, USA; 4Adult CF Center, University of Utah Cystic Fibrosis Center, Salt
Lake City, USA; 5Medicine, Penn State Milton S. Hershey Medical Center,
Hershey, USA; 6Division of Pulmonary and Critical Care Medicine, University of
Arkansas for Medical Sciences, Little Rock, USA; 7Medicine, Division of
Pulmonary and Critical Care, Larner College of Medicine, University of
Vermont, Burlington, USA; 8Pediatrics, New York Medical College, Valhalla,
USA; 9Cystic Fibrosis Center, Boston Children’s Hospital, Boston, USA; 10Clinical
Research, Calithera Biosciences, Inc., South San Francisco, USA; 11Biometrics,
Calithera Biosciences, Inc., South San Francisco, USA; 12Research, Calithera
Biosciences, Inc., South San Francisco, USA; 13Pulmonary and Critical Care
Medicine, University of Kansas Medical Center, Kansas City, USA
Background: In CF, impaired nitric oxide (NO) production may contribute
to impaired host antimicrobial defense, chronic airway infection, and
compromised pulmonary function. L-arginine (Arg) is a required substrate
of NO synthases for production of NO. Depletion of Arg by arginase, an
abundant enzyme expressed and secreted into airways by neutrophils,
contributes to NO deficiency. Clinical studies in CF patients have shown
that administration of inhaled Arg improved fractional exhaled NO (FeNO)
and trended toward improvement in FEV1. CB-280 is a potent, reversible,
oral arginase inhibitor that generates sustained increases in systemic Arg.
In preclinical studies in CF mouse models, CB-280 improved central airway
resistance and decreased lung infection. We present the first 2 dose-
escalation cohorts of an ongoing phase 1b study of CB-280 versus placebo
in adults with CF (NCT04279769; CX-280-202).
Methods: Patients were randomized 3:1 to receive CB-280 or placebo
orally over 14 days in 4 sequential dose-escalation cohorts (50, 100, 200,
400 mg twice a day; n = 8/cohort). Primary endpoint was safety and
tolerability, as assessed by adverse events (AEs), laboratory changes, ECG,
and spirometry.
Results: Results are presented for 16 patients from the first 2 cohorts (n = 6
in 50-mg cohort, n = 6 in 100-mg cohort, n = 4 placebo). Median age was
31.5 and 69% were female. Median baseline ppFEV1 was 65%; 86% of
patients were on elexacaftor/tezacaftor/ivacaftor at study entry. Treatment-
emergent AEs occurred in 3 of 12 patients treated with CB-280 and 1 of 4
patients on placebo. Treatment-related AEs in CB-280-treated patients
included Grade 1/2 dizziness and acne. No Grade 3 or greater events; dose-
limiting toxicities; serious AEs; or major adverse changes on lab
assessments, ECG, or vital signs occurred in CB-280 patients. Spirometry
showed no safety problems, and there was a positive trend in ppFEV1 with
CB-280. CB-280 exhibited linear pharmacokinetics. Steady-state Ctrough at
the 100-mg dose surpassed the IC90 for arginase inhibition in plasma,
indicating continuous target coverage. Dose-related increases in plasma
Arg were observed, with a mean 1.9-fold increase at the 100-mg dose. FeNO
showed a slightly positive trend from baseline for patients on CB-280.
Updated data will be presented.
Conclusion: In conclusion, CB-280 was well tolerated in the initial dose
cohorts of this study, with no CB-280 patients experiencing dose-limiting
toxicities. CB-280 exhibited linear pharmacokinetics, achieving continuous
target coverage in plasma at the 100-mg dose, and trended favorably in
FEV1 and FeNO parameters.
530
Novel immunomodulator ELD607 reduces neutrophilic inflammation
in βENaC and Pseudomonas aeruginosa–infected mice
S. Ahmad1, M. Sassano2, A. Ghosh2, R. Tarran2. 1Eldec Pharmaceuticals,
Chapel Hill, USA; 2Department of Cell Biology and Physiology, University of
North Carolina at Chapel Hill, Chapel Hill, USA
Background: Defective CFTR causes dehydration and acidification of the
airways, which leads to chronic bacterial infection, inflammation, and
frequent exacerbations. Repeated cycles of infection and inflammation
result in a downward spiral of injury and remodeling that ultimately leads
to bronchiectasis and respiratory failure. Therefore, management of airway
inflammation is a vital aspect of CF treatment, but other than ibuprofen,
there are no currently approved antiinflammatory drugs to treat CF
patients. Orai1 is a plasma membrane Ca2+ channel that regulates
inflammation by controlling gene expression and cytokine secretion. We
have generated ELD607, a fully optimized anti-Orai1 compound. We tested
whether inhaled ELD607 could inhibit Orai1 locally in the lungs to reduce
pulmonary inflammatory responses in mice.
Methods: To mimic CF lung disease, we initially used a epithelial sodium
channel β subunit (βENaC)-overexpressing mouse model that develops
spontaneous mucus dehydration and neutrophilic inflammation. βENaC
neonates were dosed daily intranasally with vehicle or ELD607 for 10 days
and observed for survival. Because Pseudomonas aeruginosa colonizes CF
lungs, causing lung function deterioration, wild-type C57BL/6 mice were
intranasally infected with 107 CFU/mouse P. aeruginosa and treated 1 or 24
hours later with vehicle or 0.5 mg/kg ELD607. Bronchoalveolar lavage and
whole lungs were collected 24 hours after treatment. Finally, to ensure that
the antiinflammatory effects of ELD607 did not result in suppression of an
effective immune response, mice were infected with a higher dose of
109 CFU/mouse P. aeruginosa by intranasal installation, treated with vehicle
or 1.05 mg/kg ELD607, and observed for survival.
Results: βENaC neonates treated with ELD607 exhibited less neutrophilia
and longer survival than nontreated βENaC neonates. Mice infected with P.
aeruginosa, treated 1 hour after infection with ELD607 and analyzed 24
hours after infection had lower bacterial burden than nontreated mice and
normalized neutrophil levels in the lungs. Similar results were observed
when mice were treated 24 hours after infection and analyzed 48 hours
after infection ELD607-treated mice also had significantly lower neutrophil
elastase, lactate dehydrogenase, and proinflammatory cytokine levels than
nontreated mice and longer survival than vehicle controls.
Conclusion: ELD607 significantly reduces pulmonary inflammation and
lung damage, suggesting that it may serve as a novel inhaled antiin-
flammatory immunomodulator.
Acknowledgements: Funding: NIAID R42AI155107, NHLBI R43HL144290,
NHLBI 5T32HL007106, and the NC Biotech Center.
531
Identification of a compound that mediates readthrough of CFTR
nonsense mutations by reducing eRF1 levels
D. Bedwell1, J. Sharma2, M. Du1, E. Wong3, V. Mutyam2, Y. Li2, J. Chen2,
J. Wangen4, K. Thrasher1, L. Fu5, N. Peng2, L. Tang2, K. Liu1, B. Mathew6,
B. Bostwick7, C. Augelli-Szafran6, H. Bihler3, F. Liang3, J. Mahiou3, J. Saltz3,
A. Rab8, J. Hong8, E. Sorscher8, E. Mendenhall9, C. Coppola9, K. Keeling1,
R. Green5, M. Mense3, M. Suto7, S. Rowe2. 1Biochemistry and Molecular
Genetics, University of Alabama at Birmingham, Birmingham, USA; 2Medicine,
University of Alabama at Birmingham, Birmingham, USA; 3Therapeutics Lab,
Cystic Fibrosis Foundation, Lexington, USA; 4Molecular Biology and Genetics,
Johns Hopkins University, Baltimore, USA; 5Cell, Developmental, and
Integrative Biology, University of Alabama at Birmingham, Birmingham, USA;
6
Chemistry, Southern Research, Birmingham, USA; 7High Throughput
Screening, Southern Research, Birmingham, USA; 8Pediatrics, School of
Medicine, Emory University, Atlanta, USA; 9Biological Sciences, University of
Alabama at Huntsville, Huntsville, USA
Background: Although recently developed CFTR modulator drugs can
alleviate cystic fibrosis (CF) in the majority of patients, CF patients with
mutations that form premature termination codons (PTCs) cannot benefit
from modulator therapies. PTCs terminate translation before a full-length
CF transmembrane regulator (CFTR) protein can be generated. In addition, a