# Direct Small Molecule Activation of Pro-apoptotic BAK

> **NIH NIH F30** · ALBERT EINSTEIN COLLEGE OF MEDICINE · 2021 · $51,036

## Abstract

Abstract:
 Apoptosis is a highly regulated, energy dependent form of cell death critical for the development and
homeostasis of multicellular organisms. When dysregulated, apoptosis contributes to disease states such as
cancer, autoimmunity, and neurodegeneration. Genetic and biochemical studies have established the BCL-2
family of proteins as critical regulators of mitochondrial apoptosis. Pro-apoptotic BAK is an effector of
mitochondrial apoptosis whose activation leads to mitochondrial outer membrane permeabilization (MOMP).
Upon transient interaction with a BH3-only activator protein, BAK undergoes several conformational changes
including exposure of its own BH3-domain, which allows for the formation of BAK oligomers, responsible for
MOMP. Despite its critical role in committing a cell to death, its mechanism of activation has not been fully
elucidated. The development of small molecule modulators of apoptosis has helped elucidate the role of BCL-2
proteins biochemically as well as in disease. Furthermore, anti-apoptotic inhibitors and direct and selective
BAX activators have shown promise in clinical and in vivo studies. The development of small molecule
modulators of BAK activity would aid in the elucidation of its activation mechanism as well as demonstrate the
therapeutic potential of pharmacologic control of BAK. Here, we propose to develop small molecule activators
of BAK from fragment 7H8 identified in an NMR-based fragment screen. Our specific aims are 1) to design and
synthesize small molecule analogs of 7H8 targeting BAK and determine their activation mechanism and 2) to
evaluate the biochemical and cellular activity of small molecules. To accomplish our first aim, we will use
HSQC-NMR and computational docking data to virtually design and chemically synthesize elaborated 7H8
analogs. We will then determine their affinity and specificity using microscale thermophoresis and fluorescence
polarization binding assays. We will use hydrogen-deuterium exchange mass spectrometry to determine the
conformational changes associated with BAK activation. We will accomplish our second aim by assessing
small molecule activity in vitro using liposomal fluorescence release, isolated mitochondrial cytochrome c
release, and oligomerization and conformational change assays. We will then assess activity in genetically
modified mouse embryonic fibroblasts to assess for cellular activity and specificity. By realizing these aims, this
proposal will advance our understanding of the activation mechanism of pro-apoptotic BAK and demonstrate a
new paradigm for pharmacologic induction of apoptosis through BAK activation. Under the guidance of my
mentors, I will be able to accomplish these goals and further expand our knowledge of BAK's role in apoptosis
and human disease while gaining skills and knowledge in structural biology, biophysics, cellular biology,
biochemistry, and medicinal chemistry. I will expand my training through attending scientific meetings an...

## Key facts

- **NIH application ID:** 10076802
- **Project number:** 5F30CA228453-04
- **Recipient organization:** ALBERT EINSTEIN COLLEGE OF MEDICINE
- **Principal Investigator:** Adam Zoltan Spitz
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $51,036
- **Award type:** 5
- **Project period:** 2019-01-01 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10076802

## Citation

> US National Institutes of Health, RePORTER application 10076802, Direct Small Molecule Activation of Pro-apoptotic BAK (5F30CA228453-04). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10076802. Licensed CC0.

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