{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/88246"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/88246","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Improving the process synthesis, potency, and pharmacokinetics of the anticancer compound PAC-1","abstract":"Embargo set by: Seth Robbins for item 89526 Lift date: 2017-09-29T21:08:35Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","abstract_html":"Embargo set by: Seth Robbins for item 89526 Lift date: 2017-09-29T21:08:35Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","abstract_has_math":false,"creators":["Roth, Howard Steven"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemistry","degree_department":null,"school":null,"contributors":["Hergenrother, Paul J.","Katzenellenbogen, John A.","Mitchell, Douglas A.","Fan, Timothy M."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-09-29T21:02:49Z","date_published":"2015-09-29T21:02:49Z","updated_at":"2026-07-22T22:26:31Z","subjects":["first procaspase activating compound (PAC-1)","procaspase-3","zinc"],"languages":["en"],"rights":["Copyright 2015 Howard Roth"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/88246","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Hergenrother, Paul J.","Katzenellenbogen, John A.","Mitchell, Douglas A.","Fan, Timothy M."]},{"key":"dc:creator","label":"Author","values":["Roth, Howard Steven"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2015-09-29T21:02:49Z","2017-09-30T09:15:30Z","2015-08","2015-06-17","2015-8"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["first procaspase activating compound (PAC-1)","procaspase-3","zinc"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2015 Howard Roth"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/88246"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Embargo set by: Seth Robbins for item 89526 Lift date: 2017-09-29T21:08:35Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 89526 on 2017-09-30T09:15:30Z.","PAC-1 is an ortho-hydroxy-N-acylhydrazone that induces apoptosis by chelation of antiapoptotic labile zinc, relieving zinc-mediated inhibition of procaspase-3. Favorable results from cell culture and in vivo experiments with PAC-1 and its derivative S-PAC-1 have indicated the potential for translation to the clinic. However, several challenges exist in using PAC-1 in these experiments. This report describes the attempts to improve upon the process synthesis, potency, pharmacokinetics, and safety of PAC-1. The synthetic route to access PAC-1 was successful for the generation of batches of approximately 25 grams, but adaptation to multi-kilogram scale, as would be necessary for a human clinical trial, was not feasible using the existing route. In particular, the route included three chromatographic purification steps, as well as the use of anhydrous hydrazine. Optimization studies indicated that the active pharmaceutical ingredient could be synthesized without purification of any intermediates, and hydrazine monohydrate could be used in place of the anhydrous reagent. A batch of 155 grams of PAC-1 was synthesized using the improved route, and the route was scaled up to produce a batch of greater than 10 kilograms by a contract research organization. The lessons learned through this optimization effort were general and should be applicable if an alternative PAC-1 derivative is identified. In an effort to discover PAC-1 derivatives with improved potency, a combinatorial library of 837 compounds was synthesized. Each of 31 hydrazides was condensed with each of 27 aldehydes. Because standard techniques for isolation and purification of products are impractical for libraries of this size, a solid-phase purification strategy was employed that yielded the library members with an average purity of 91%. Compounds were screened in cell culture, and six potent hits were identified. One of these compounds, B-PAC-1, was studied further in primary isolates from leukemia patients, and animal experiments are currently in progress. An additional challenge is the relatively rapid clearance of PAC-1 from circulation. Many sites of metabolism exist, giving metabolites that arise from oxidative N-dealkylation, olefin oxidation, and arene oxidation. Several derivatives were synthesized containing modifications that prevent the formation of these metabolites. The compounds were evaluated in cell culture, liver microsomes, and mice, and four lead compounds were identified. Pharmacokinetic analysis indicated that each of these four compounds displayed extended elimination half-lives compared to PAC-1 and S-PAC-1, making these compounds viable candidates as next-generation PAC-1 derivatives. One of the most significant challenges in working with PAC-1 in vivo is the neuroexcitation observed upon treatment with elevated doses of compound. Preliminary experiments suggested that the neurotoxicity was not related to the ability of the compound to bind zinc. In order to confirm these results and attempt to determine the portion of the molecule responsible for neurotoxicity, a series of compounds was designed and synthesized containing systematic modifications to PAC-1. Evaluation in cell culture and in vitro further defined the essential nature of the ortho-hydroxy-N-acylhydrazone for activity. Evaluation of the compounds in mice confirmed previous experiments that the neurotoxicity is most likely not a result of metal chelation, but no further conclusions could be drawn from these experiments, and further study will be necessary to fully define the observed neurotoxicity.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2017-08-01","The student, Howard Roth, accepted the attached license on 2015-06-15 at 10:15.","The student, Howard Roth, submitted this Dissertation for approval on 2015-06-15 at 10:18.","This Dissertation was approved for publication on 2015-06-17 at 10:45.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8274 on 2015-09-29 at 15:05:11","Made available in DSpace on 2015-09-29T21:02:49Z (GMT). No. of bitstreams: 2 ROTH-DISSERTATION-2015.pdf: 13544001 bytes, checksum: b134b687899abee227ae8dad3e7a9e5b (MD5) LICENSE.txt: 4208 bytes, checksum: b8728bf14571369ae7733c13d4cac1a9 (MD5) Previous issue date: 2015-06-17","Embargo set by: Seth Robbins for item 89526 Lift date: 2017-09-29T21:03:28Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Improving the process synthesis, potency, and pharmacokinetics of the anticancer compound PAC-1"]}]}],"canonical_facts":{"dc:contributor":["Hergenrother, Paul J.","Katzenellenbogen, John A.","Mitchell, Douglas A.","Fan, Timothy M."],"dc:creator":["Roth, Howard Steven"],"dc:date":["2015-09-29T21:02:49Z","2017-09-30T09:15:30Z","2015-08","2015-06-17","2015-8"],"dc:description":["Embargo set by: Seth Robbins for item 89526 Lift date: 2017-09-29T21:08:35Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 89526 on 2017-09-30T09:15:30Z.","PAC-1 is an ortho-hydroxy-N-acylhydrazone that induces apoptosis by chelation of antiapoptotic labile zinc, relieving zinc-mediated inhibition of procaspase-3. Favorable results from cell culture and in vivo experiments with PAC-1 and its derivative S-PAC-1 have indicated the potential for translation to the clinic. However, several challenges exist in using PAC-1 in these experiments. This report describes the attempts to improve upon the process synthesis, potency, pharmacokinetics, and safety of PAC-1. The synthetic route to access PAC-1 was successful for the generation of batches of approximately 25 grams, but adaptation to multi-kilogram scale, as would be necessary for a human clinical trial, was not feasible using the existing route. In particular, the route included three chromatographic purification steps, as well as the use of anhydrous hydrazine. Optimization studies indicated that the active pharmaceutical ingredient could be synthesized without purification of any intermediates, and hydrazine monohydrate could be used in place of the anhydrous reagent. A batch of 155 grams of PAC-1 was synthesized using the improved route, and the route was scaled up to produce a batch of greater than 10 kilograms by a contract research organization. The lessons learned through this optimization effort were general and should be applicable if an alternative PAC-1 derivative is identified. In an effort to discover PAC-1 derivatives with improved potency, a combinatorial library of 837 compounds was synthesized. Each of 31 hydrazides was condensed with each of 27 aldehydes. Because standard techniques for isolation and purification of products are impractical for libraries of this size, a solid-phase purification strategy was employed that yielded the library members with an average purity of 91%. Compounds were screened in cell culture, and six potent hits were identified. One of these compounds, B-PAC-1, was studied further in primary isolates from leukemia patients, and animal experiments are currently in progress. An additional challenge is the relatively rapid clearance of PAC-1 from circulation. Many sites of metabolism exist, giving metabolites that arise from oxidative N-dealkylation, olefin oxidation, and arene oxidation. Several derivatives were synthesized containing modifications that prevent the formation of these metabolites. The compounds were evaluated in cell culture, liver microsomes, and mice, and four lead compounds were identified. Pharmacokinetic analysis indicated that each of these four compounds displayed extended elimination half-lives compared to PAC-1 and S-PAC-1, making these compounds viable candidates as next-generation PAC-1 derivatives. One of the most significant challenges in working with PAC-1 in vivo is the neuroexcitation observed upon treatment with elevated doses of compound. Preliminary experiments suggested that the neurotoxicity was not related to the ability of the compound to bind zinc. In order to confirm these results and attempt to determine the portion of the molecule responsible for neurotoxicity, a series of compounds was designed and synthesized containing systematic modifications to PAC-1. Evaluation in cell culture and in vitro further defined the essential nature of the ortho-hydroxy-N-acylhydrazone for activity. Evaluation of the compounds in mice confirmed previous experiments that the neurotoxicity is most likely not a result of metal chelation, but no further conclusions could be drawn from these experiments, and further study will be necessary to fully define the observed neurotoxicity.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2017-08-01","The student, Howard Roth, accepted the attached license on 2015-06-15 at 10:15.","The student, Howard Roth, submitted this Dissertation for approval on 2015-06-15 at 10:18.","This Dissertation was approved for publication on 2015-06-17 at 10:45.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8274 on 2015-09-29 at 15:05:11","Made available in DSpace on 2015-09-29T21:02:49Z (GMT). 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