Date of Award

Fall 12-2025

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Pharmaceutical Sciences

First Advisor

Kamaljit Kaur

Second Advisor

Rachita Sumbria

Third Advisor

Jennifer Totonchy

Abstract

Amylin receptor antagonist AC253 is a 24-amino acid peptide that provides neuroprotection against amyloid beta (Aβ)-induced cell death and toxicity. Additionally, AC253 has been shown to improve spatial memory in mouse models of Alzheimer’s disease (AD). Screening of short overlapping fragments (12-mer) of AC253 showed that N-terminal residues were responsible for high affinity binding to the amylin receptor (AMY3). Both AC253 and R5 (a 12-mer fragment) have a short half-life in human serum and are hydrophilic peptides with a low log P (-1.2 for AC253) and are not optimized for brain uptake. From the structure-activity relationship studies, we have designed a 16-mer analog (R16) from the N-terminal region of AC253 as a novel AmyR antagonist for better activity and blood-brain permeability. The goal of this study is to design and synthesize a novel neuroprotective analog of AC253 called R16 for enhanced blood-brain barrier targeting. R16 was synthesized using solid-phase peptide synthesis, purified and characterized by MALDI-TOF mass spectrometry and reversed-phase HPLC. The proteolytic stability of R16 was evaluated in human and mouse sera to determine the half-lives and proteolytically labile sites. The half-life of R16 was found to be 8.98 hours and 0.47 hours in human and mouse serum, respectively. The N-terminal of R16 was found to be accessible to proteolytic cleavage by serum proteases. Therefore, blocking N-terminal of R16 with a blood brain targeting peptide Angiopep-2 led to the design of a proteolytic stable blood-brain barrier targeting analog of R16, namely Angiopep2- R16 conjugate. The conjugate was synthesized, purified and characterized using reversed-phase HPLC and mass spectrometry, and was found to be >95% pure. The conjugate will be further studied in the future to test its pharmacokinetics, stability, and brain activity.

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Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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