One of the major problems of the common chemotherapeutic drugs are the resistance faced as well as significant side effects.

BaX, a protein remains in cytosol & on initiation of apoptosis move to mitochondria and activates caspases through cytochrome c release.

Two of the major activators studied are BAM7 and BTSA1.

Through Docking analysis, we found BTSA1 and BAM7 affect Alpha 2 and Alpha 6 helices directly

v BAM7 relies on a strong, optimal alkyl bonds to anchor itself precisely, complemented by a broad set of interactions across the pocket, conferring stability

v The three strong alkyl bonds are at Ala 34 and Met 137

v BTSA1 serves as a characteristic activator creating a more powerful hydrophobic interaction network.

v  Highlighted by its dual engagement with ARG 134 and pi alkyl bond with lys21.

 

In a 50ns MD simulation, The BAM7-BAX showed lower fluctuations than BTSA1-BaX complex that showed 2 distinct states, that’s why we decided to extend the run to 100ns to validate.

�� Binding at  alpha 2 and alpha 6 residues lead to RMSF peaks (with BTSA1 showing higher peaks), showing strong rearrangements

�� The activation is proved by peak at BH3 helix and alpha 5 loops.

��Interestingly, the peak at around residue 57 and 96 in BTSA1 starts very low but gradually grows and reaches peak at arounf 50-55 ns before sunsiding

�� BAM7 starts off with high flexibility and the peak suppresses a bit in the final frame. Proving that BTSA1 is more potent

Pearson’s cross coefficient

• In BAM7 Diagonal line (self-correlation) is strong, rest are weak, Suggests a more localized and less cooperative residue motion
• BTSA1 Higher density of both pink (positive) and green (negative) regions → indicates greater cooperativity and inter-domain communication between residue groups.

In-Silico Toxicity Assay

��Both show hepatotoxicity & nephrotoxicity, BTSA1 has a higher probability (91.8 & 89.1% vs. 85.5 &79.9%),showing respiratory toxicity, slightly higher clearance and bio-availability. (admetSAR)

��Bam7 and btsa1 both show good drug-likeness and no significant off-target interactions in SwissADME. (Complete 0 in BAM7 and 0.106% ADORA1 and ADORA2B in BTSA1)

��One Problem Faced by BTSA1 is carcinogenicity as shown in admetSAR, but it needs clinal validation

�� BTSA1 drives drastic conformational changes, with a higher toxicity profile

We conclude that It can be suitable for senescence-targeted therapies, hematologic malignancies or higher stages of metastasis where apoptosis induction is desirable.

�� BAM7, while still inducing apoptosis, appears to be weak but controlled modulator of BAX, with fewer off-target effects and

 �� It can be used in long term therapies, pulsed therapies, synergistic treatment or in lower stages of cancer.

 

We collected data related to BAM7 sensitivity from GDSC and depMAP and made a rule based classifier on the sensitivity of BAM7 across 488 cell lines.

This model will serve as a pioneer to study the BAM7 sensitivity across cell lines under one roof.