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.
","abstract_html":"<p>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.</p>","abstract_has_math":false,"creators":["Bellili, Riad"],"institution":null,"degree_name":"Master of Science (MS)","degree_level":"Thesis","degree_discipline":"Pharmaceutical Sciences","degree_department":null,"school":null,"contributors":["Kamaljit Kaur","Rachita Sumbria","Jennifer Totonchy"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-12-01T08:00:00Z","date_published":"2025-12-01T08:00:00Z","updated_at":"2026-07-24T01:38:43Z","subjects":["Other Pharmacy and Pharmaceutical Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/41","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Kamaljit Kaur","Rachita Sumbria","Jennifer Totonchy"]},{"key":"dc:creator","label":"Author","values":["Bellili, Riad"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2027-12-23T08:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Pharmaceutical Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science (MS)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Other Pharmacy and Pharmaceutical Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/41"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.
"]},{"key":"dc:source","label":"Dc Source","values":["Bellili, R. Design and Synthesis of a Novel Neuroprotective Brain-Targeting Amylin Receptor Antagonist. [master’s thesis]. Irvine, CA: Chapman University; 2025. https://doi.org/10.36837/chapman.000715"]},{"key":"dc:title","label":"Title","values":["Design and Synthesis of a Novel Neuroprotective Brain-Targeting Amylin Receptor Antagonist"]}]}],"canonical_facts":{"dc:contributor":["Kamaljit Kaur","Rachita Sumbria","Jennifer Totonchy"],"dc:creator":["Bellili, Riad"],"dc:date.available":["2027-12-23T08:00:00Z"],"dc:description.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.
"],"dc:identifier":["https://digitalcommons.chapman.edu/pharmaceutical_sciences_theses/41"],"dc:source":["Bellili, R. Design and Synthesis of a Novel Neuroprotective Brain-Targeting Amylin Receptor Antagonist. [master’s thesis]. Irvine, CA: Chapman University; 2025. https://doi.org/10.36837/chapman.000715"],"dc:subject":["Other Pharmacy and Pharmaceutical Sciences"],"dc:title":["Design and Synthesis of a Novel Neuroprotective Brain-Targeting Amylin Receptor Antagonist"],"thesis:degree_discipline":["Pharmaceutical Sciences"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["Master of Science (MS)"]},"updated_at":"2026-07-24T01:38:43Z"}