# Allosteric Pharmacologic Chaperones for alpha-1 Antitrypsin Mutants

> **NIH NIH F31** · SCRIPPS RESEARCH INSTITUTE, THE · 2023 · $34,694

## Abstract

Project Summary/Abstract
The major deficiency E342K mutant or “Z-variant” of the abundant serum antiprotease Alpha-1 antitrypsin
(AAT) is responsible for the vast majority of morbidity and mortality associated with Alpha-1 antitrypsin
deficiency (AATD), a leading cause of hereditary lung and liver disease affecting millions of patients globally.
This missasembly-prone variant is known to be highly polymerogenic, owing to a widened -sheet A domain
which predisposes the AAT-Z monomer to form cytotoxic loop-sheet oligomers. These toxic oligomers
accumulate in producing hepatocytes leading to chronic liver disease, and build up extracellularly leading to
both gain-of-toxic function in the lung with a concomitant loss of serum AAT-Z antiprotease activity which leads
to proteolytic destruction of lung parenchyma by neutrophil proteases like elastase. Crystal structures of
monomeric AAT-Z have been studied and previous mutational analyses have demonstrated the capacity for
space-filling mutations within surface-accessible hydrophobic pockets on this protein to prevent polymerization
without abrogating antiprotease activity. It has thus been hypothesized that small molecules could be
discovered which act as pharmacological chaperones, preventing AAT-Z polymer formation while permitting
native antiprotease activity of the stabilized monomer, serving to ameliorate the multiorgan injury (including
lung pathology) associated with AATD. While stabilizing ligands for AAT-Z have previously been reported,
these ligands universally fail to permit the native antiprotease activity of AAT-Z. In this training proposal, I
propose to use Fully-Functionalized Fragment” (FFF) substructures along with photo-crosslinking, affinity
chromatography and tandem mass-spectrometry in Aim 1 to identify the small molecule sites on the AAT-Z
monomer accessible to binding by drug-like substructures. In Aim 2 I will develop a novel screening assay that
I conceived of for identifying AAT-Z stabilizing ligands or pharmacologic chaperones that prevent RCL insertion
while permitting antiprotease activity of ligand-bound AAT-Z. In Aim 3 I will employ a cell-based phenotypic
assay already developed and validated by our collaborators in the Balch Lab to simultaneously evaluate the
capacity for screening hits to restore functional monomeric AAT-Z secretion efficiency, while reducing
intracellular oligomers in cultured hepatocyte and pulmonary cell models of AATD. This project will afford a
multidisciplinary training experience with guidance from experts in the fields of protein misfolding biology,
chemical proteomics, high-throughput assay development, as well as AATD patient treatment. Through the
research proposed herein, I will develop a robust expertise in state-of-the-art methods for proteomic analysis,
screening assay optimization, and applications of mammalian cell models of disease. These valuable
discovery-oriented research competencies, together with my previous training background...

## Key facts

- **NIH application ID:** 10633070
- **Project number:** 5F31GM145186-02
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** Adrian M. Guerrero
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $34,694
- **Award type:** 5
- **Project period:** 2022-02-01 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10633070

## Citation

> US National Institutes of Health, RePORTER application 10633070, Allosteric Pharmacologic Chaperones for alpha-1 Antitrypsin Mutants (5F31GM145186-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10633070. Licensed CC0.

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