{"title":"Bacterial Cytochrome P450 Catalyzed Macrocyclization of Ribosomal Peptides.","authors":"Jing Liu, Runze Liu, Bei-Bei He, Xiaoqian Lin, Longcheng Guo, Gengfan Wu, Yong-Xin Li","doi":"10.1021/acsbiomedchemau.4c00080","DOIUrl":"10.1021/acsbiomedchemau.4c00080","url":null,"abstract":"<p><p>Macrocyclization is a vital process in the biosynthesis of ribosomally synthesized and post-translationally modified peptides (RiPPs), significantly enhancing their structural diversity and biological activity. Universally found in living organisms, cytochrome P450 enzymes (P450s) are versatile catalysts that facilitate a wide array of chemical transformations and have recently been discovered to contribute to the expansion and complexity of the chemical spectrum of RiPPs. Particularly, P450-catalyzed biaryl-bridged RiPPs, characterized by highly modified structures, represent an intriguing but underexplored class of natural products, as demonstrated by the recent discovery of tryptorubin A, biarylitide and cittilin. These P450 enzymes demonstrate their versatility by facilitating peptide macrocyclization through the formation of carbon-carbon (C-C), carbon-nitrogen (C-N) and ether bonds between the side chains of tyrosine (Tyr), tryptophan (Trp) and histidine (His). This Review briefly highlights the latest progress in P450-catalyzed macrocyclization within RiPP biosynthesis, resulting in the generation of structurally complex RiPPs. These findings have expedited the discovery and detailed analysis of new P450s engaged in RiPP biosynthetic pathways.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"268-279"},"PeriodicalIF":3.8,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659900/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Bacterial Cytochrome P450 Catalyzed Macrocyclization of Ribosomal Peptides","authors":"Jing Liu, Runze Liu, Bei-Bei He, Xiaoqian Lin, Longcheng Guo, Gengfan Wu and Yong-Xin Li*, ","doi":"10.1021/acsbiomedchemau.4c0008010.1021/acsbiomedchemau.4c00080","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00080https://doi.org/10.1021/acsbiomedchemau.4c00080","url":null,"abstract":"<p >Macrocyclization is a vital process in the biosynthesis of ribosomally synthesized and post-translationally modified peptides (RiPPs), significantly enhancing their structural diversity and biological activity. Universally found in living organisms, cytochrome P450 enzymes (P450s) are versatile catalysts that facilitate a wide array of chemical transformations and have recently been discovered to contribute to the expansion and complexity of the chemical spectrum of RiPPs. Particularly, P450-catalyzed biaryl-bridged RiPPs, characterized by highly modified structures, represent an intriguing but underexplored class of natural products, as demonstrated by the recent discovery of tryptorubin A, biarylitide and cittilin. These P450 enzymes demonstrate their versatility by facilitating peptide macrocyclization through the formation of carbon–carbon (C–C), carbon–nitrogen (C–N) and ether bonds between the side chains of tyrosine (Tyr), tryptophan (Trp) and histidine (His). This Review briefly highlights the latest progress in P450-catalyzed macrocyclization within RiPP biosynthesis, resulting in the generation of structurally complex RiPPs. These findings have expedited the discovery and detailed analysis of new P450s engaged in RiPP biosynthetic pathways.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"268–279 268–279"},"PeriodicalIF":3.8,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00080","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142843254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-20eCollection Date: 2024-12-18DOI: 10.1021/acsbiomedchemau.4c00089
Madison Knapp, Minshik Jo, Courtney L Henthorn, Marley Brimberry, Andrew D Gnann, Daniel P Dowling, Jennifer Bridwell-Rabb
{"title":"Chlorophyllase from <i>Arabidopsis thaliana</i> Reveals an Emerging Model for Controlling Chlorophyll Hydrolysis.","authors":"Madison Knapp, Minshik Jo, Courtney L Henthorn, Marley Brimberry, Andrew D Gnann, Daniel P Dowling, Jennifer Bridwell-Rabb","doi":"10.1021/acsbiomedchemau.4c00089","DOIUrl":"10.1021/acsbiomedchemau.4c00089","url":null,"abstract":"<p><p>Chlorophyll (Chl) is one of Nature's most complex pigments to biosynthesize and derivatize. This pigment is vital for survival and also paradoxically toxic if overproduced or released from a protective protein scaffold. Therefore, along with the mass production of Chl, organisms also invest in mechanisms to control its degradation and recycling. One important enzyme that is involved in these latter processes is chlorophyllase. This enzyme is employed by numerous photosynthetic organisms to hydrolyze the phytol tail of Chl. Although traditionally thought to catalyze the first step of Chl degradation, recent work suggests that chlorophyllase is instead employed during times of abiotic stress or conditions that produce reactive oxygen species. However, the molecular details regarding how chlorophyllases are regulated to function under such conditions remain enigmatic. Here, we investigate the <i>Arabidopsis thaliana</i> chlorophyllase isoform <i>At</i>CLH2 using site-directed mutagenesis, mass spectrometry, dynamic light scattering, size-exclusion multiangle light scattering, and both steady-state enzyme kinetic and thermal stability measurements. Through these experiments, we show that <i>At</i>CLH2 exists as a monomer in solution and contains two disulfide bonds. One disulfide bond putatively maps to the active site, whereas the other links two N-terminal Cys residues together. These disulfide bonds are cleaved by chemical or chemical and protein-based reductants, respectively, and are integral to maintaining the activity, stability, and substrate scope of the enzyme. This work suggests that Cys residue oxidation in chlorophyllases is an emerging regulatory strategy for controlling the hydrolysis of Chl pigments.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"353-370"},"PeriodicalIF":3.8,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659893/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-20DOI: 10.1021/acsbiomedchemau.4c0008910.1021/acsbiomedchemau.4c00089
Madison Knapp, Minshik Jo, Courtney L. Henthorn, Marley Brimberry, Andrew D. Gnann, Daniel P. Dowling and Jennifer Bridwell-Rabb*,
{"title":"Chlorophyllase from Arabidopsis thaliana Reveals an Emerging Model for Controlling Chlorophyll Hydrolysis","authors":"Madison Knapp, Minshik Jo, Courtney L. Henthorn, Marley Brimberry, Andrew D. Gnann, Daniel P. Dowling and Jennifer Bridwell-Rabb*, ","doi":"10.1021/acsbiomedchemau.4c0008910.1021/acsbiomedchemau.4c00089","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00089https://doi.org/10.1021/acsbiomedchemau.4c00089","url":null,"abstract":"<p >Chlorophyll (Chl) is one of Nature’s most complex pigments to biosynthesize and derivatize. This pigment is vital for survival and also paradoxically toxic if overproduced or released from a protective protein scaffold. Therefore, along with the mass production of Chl, organisms also invest in mechanisms to control its degradation and recycling. One important enzyme that is involved in these latter processes is chlorophyllase. This enzyme is employed by numerous photosynthetic organisms to hydrolyze the phytol tail of Chl. Although traditionally thought to catalyze the first step of Chl degradation, recent work suggests that chlorophyllase is instead employed during times of abiotic stress or conditions that produce reactive oxygen species. However, the molecular details regarding how chlorophyllases are regulated to function under such conditions remain enigmatic. Here, we investigate the <i>Arabidopsis thaliana</i> chlorophyllase isoform <i>At</i>CLH2 using site-directed mutagenesis, mass spectrometry, dynamic light scattering, size-exclusion multiangle light scattering, and both steady-state enzyme kinetic and thermal stability measurements. Through these experiments, we show that <i>At</i>CLH2 exists as a monomer in solution and contains two disulfide bonds. One disulfide bond putatively maps to the active site, whereas the other links two N-terminal Cys residues together. These disulfide bonds are cleaved by chemical or chemical and protein-based reductants, respectively, and are integral to maintaining the activity, stability, and substrate scope of the enzyme. This work suggests that Cys residue oxidation in chlorophyllases is an emerging regulatory strategy for controlling the hydrolysis of Chl pigments.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"353–370 353–370"},"PeriodicalIF":3.8,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00089","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142843024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-19DOI: 10.1021/acsbiomedchemau.4c0008810.1021/acsbiomedchemau.4c00088
Mochen Dong, Zhuoyun Chen, Yuan He, Rémi Zallot and Yi Jin*,
{"title":"Bioinformatics-Facilitated Identification of Novel Bacterial Sulfoglycosidases That Hydrolyze 6-Sulfo-N-acetylglucosamine","authors":"Mochen Dong, Zhuoyun Chen, Yuan He, Rémi Zallot and Yi Jin*, ","doi":"10.1021/acsbiomedchemau.4c0008810.1021/acsbiomedchemau.4c00088","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00088https://doi.org/10.1021/acsbiomedchemau.4c00088","url":null,"abstract":"<p >Glycan sulfation is a widespread postglycosylation modification crucial for modulating biological functions including cellular adhesion, signaling, and bacterial colonization. 6-Sulfo-β-GlcNAcases are a class of enzyme that alters sulfation patterns. Such changes in sulfation patterns are linked to diseases such as bowel inflammation, colitis, and cancer. Despite their significance, 6-sulfo-β-GlcNAcases, which cleave β-linked 6-sulfo-<i>N</i>-acetylglucosamine (6S-GlcNAc), have been but rarely identified. This scarcity results mainly from the short, diverse, and distinctive sulfate-binding motifs required for recognition of the 6-sulfate group in 6S-GlcNAc in addition to the conserved GH20 family features. In this study, we discovered 6-sulfo-β-GlcNAcases and assigned two novel sulfate-binding motifs by the use of comparative genomics, structural predictions, and activity-based screening. Our findings expand the known microbiota capable of degrading sulfated glycans and add significant enzymes to the tool kit for analysis and synthesis of sulfated oligosaccharides.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"342–352 342–352"},"PeriodicalIF":3.8,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00088","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142842948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-19eCollection Date: 2024-12-18DOI: 10.1021/acsbiomedchemau.4c00088
Mochen Dong, Zhuoyun Chen, Yuan He, Rémi Zallot, Yi Jin
{"title":"Bioinformatics-Facilitated Identification of Novel Bacterial Sulfoglycosidases That Hydrolyze 6-Sulfo-<i>N</i>-acetylglucosamine.","authors":"Mochen Dong, Zhuoyun Chen, Yuan He, Rémi Zallot, Yi Jin","doi":"10.1021/acsbiomedchemau.4c00088","DOIUrl":"10.1021/acsbiomedchemau.4c00088","url":null,"abstract":"<p><p>Glycan sulfation is a widespread postglycosylation modification crucial for modulating biological functions including cellular adhesion, signaling, and bacterial colonization. 6-Sulfo-β-GlcNAcases are a class of enzyme that alters sulfation patterns. Such changes in sulfation patterns are linked to diseases such as bowel inflammation, colitis, and cancer. Despite their significance, 6-sulfo-β-GlcNAcases, which cleave β-linked 6-sulfo-<i>N</i>-acetylglucosamine (6S-GlcNAc), have been but rarely identified. This scarcity results mainly from the short, diverse, and distinctive sulfate-binding motifs required for recognition of the 6-sulfate group in 6S-GlcNAc in addition to the conserved GH20 family features. In this study, we discovered 6-sulfo-β-GlcNAcases and assigned two novel sulfate-binding motifs by the use of comparative genomics, structural predictions, and activity-based screening. Our findings expand the known microbiota capable of degrading sulfated glycans and add significant enzymes to the tool kit for analysis and synthesis of sulfated oligosaccharides.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"342-352"},"PeriodicalIF":3.8,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659886/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Ex Vivo Delivery of mRNA to Immune Cells via a Nonendosomal Route Obviates the Need for Nucleoside Modification","authors":"Bartika Ghoshal, Debajyoti Chakraborty, Manish Nag, Raghavan Varadarajan and Siddharth Jhunjhunwala*, ","doi":"10.1021/acsbiomedchemau.4c0005710.1021/acsbiomedchemau.4c00057","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00057https://doi.org/10.1021/acsbiomedchemau.4c00057","url":null,"abstract":"<p >Base modification and the use of lipid nanoparticles are thought to be essential for efficient in vivo delivery and expression of mRNA. However, for ex vivo immune cell engineering, the need for either of the two is unclear. Previous reports have suggested that nucleic acids may be efficiently delivered to immune cells ex vivo, through a nonendosomal delivery route, but the need for base modification has not been determined. Herein, we demonstrate that when a nonendosomal delivery method is used, unmodified mRNA performs equally well to the commonly used base-modified mRNA, including the <i>N</i><sup>1</sup> methyl pseudouridine modification, in terms of protein expression and inflammatory response in cells. However, if an endosomal delivery route is used, then <i>N</i><sup>1</sup> methyl pseudouridine modification is necessary for high expression and low inflammatory response, as demonstrated by others as well. Overall, we show that nonendosomal mRNA delivery renders nucleoside modifications nonessential and that unmodified mRNA combined with nonendosomal delivery route may be used for efficient ex vivo mRNA-based engineering of immune cells.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"291–299 291–299"},"PeriodicalIF":3.8,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00057","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142842911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Ex Vivo Delivery of mRNA to Immune Cells via a Nonendosomal Route Obviates the Need for Nucleoside Modification.","authors":"Bartika Ghoshal, Debajyoti Chakraborty, Manish Nag, Raghavan Varadarajan, Siddharth Jhunjhunwala","doi":"10.1021/acsbiomedchemau.4c00057","DOIUrl":"10.1021/acsbiomedchemau.4c00057","url":null,"abstract":"<p><p>Base modification and the use of lipid nanoparticles are thought to be essential for efficient in vivo delivery and expression of mRNA. However, for ex vivo immune cell engineering, the need for either of the two is unclear. Previous reports have suggested that nucleic acids may be efficiently delivered to immune cells ex vivo, through a nonendosomal delivery route, but the need for base modification has not been determined. Herein, we demonstrate that when a nonendosomal delivery method is used, unmodified mRNA performs equally well to the commonly used base-modified mRNA, including the <i>N</i> <sup>1</sup> methyl pseudouridine modification, in terms of protein expression and inflammatory response in cells. However, if an endosomal delivery route is used, then <i>N</i> <sup>1</sup> methyl pseudouridine modification is necessary for high expression and low inflammatory response, as demonstrated by others as well. Overall, we show that nonendosomal mRNA delivery renders nucleoside modifications nonessential and that unmodified mRNA combined with nonendosomal delivery route may be used for efficient ex vivo mRNA-based engineering of immune cells.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"291-299"},"PeriodicalIF":3.8,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659889/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-05eCollection Date: 2024-12-18DOI: 10.1021/acsbiomedchemau.4c00087
Noah M Moriarty, Annaleigh M Benton, Lauren E Gartenhaus, Andrew R Nelson, Haley A Harper, Carli J McMahan, Bennett D Elzey, Jason A Hanna, Elizabeth I Parkinson
{"title":"Design, Synthesis, and Evaluation of Trihalomethyl Ketone Derivatives of Neocarzilin A as Improved Antimetastatic Agents.","authors":"Noah M Moriarty, Annaleigh M Benton, Lauren E Gartenhaus, Andrew R Nelson, Haley A Harper, Carli J McMahan, Bennett D Elzey, Jason A Hanna, Elizabeth I Parkinson","doi":"10.1021/acsbiomedchemau.4c00087","DOIUrl":"10.1021/acsbiomedchemau.4c00087","url":null,"abstract":"<p><p>Vesicle Amine Transport-1 (VAT1) is a protein that is overexpressed in many cancers, including breast cancer, glioblastoma, and angiosarcoma. High VAT1 expression correlates with poor overall survival, and genetic knockout models of VAT1 indicate potent antimigratory activity, suggesting that VAT1 is a promising antimetastasis target. Recently, the natural product neocarzilin A (NCA) from <i>Streptomyces carzinostaticus</i> was reported to be the first validated small-molecule inhibitor of VAT1, having strong activity in metastasis models of angiosarcoma and breast cancer. While knockdown of VAT1 has no effect on cell viability, NCA has significant cytotoxicity, suggesting that NCA is not selective for VAT1. Additionally, NCA has poor aqueous solubility, making <i>in vivo</i> administration of NCA challenging and thus limiting its therapeutic potential. Here, we report the design, synthesis, bioactivity, and pharmacokinetics of novel NCA derivatives with improved drug-like properties. Specifically, we have developed derivatives with altered warheads, replacing chlorines on the trichloroketone with fluorines. Using a modified synthetic route, we accessed NCA derivatives with greater than 25-fold improvements in solubility and 30-fold improvements in the antimigratory to antiproliferative bioactivity ratio. The two best derivatives, along with the parent, were analyzed for oral bioavailability, with the two more soluble derivatives showing greatly improved bioavailability. Overall, these studies have resulted in the development of VAT1 inhibitors with improved properties, which will enable further study of the pharmacological inhibition of VAT1 as an antimetastatic strategy. Additionally, these studies provide insights into novel trihalomethyl ketone warheads and identify chlorodifluoroketone as a potent and selective new warhead.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"331-341"},"PeriodicalIF":3.8,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659896/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
ACS Bio & Med Chem AuPub Date : 2024-11-05DOI: 10.1021/acsbiomedchemau.4c0008710.1021/acsbiomedchemau.4c00087
Noah M. Moriarty, Annaleigh M. Benton, Lauren E. Gartenhaus, Andrew R. Nelson, Haley A. Harper, Carli J. McMahan, Bennett D. Elzey, Jason A. Hanna* and Elizabeth I. Parkinson*,
{"title":"Design, Synthesis, and Evaluation of Trihalomethyl Ketone Derivatives of Neocarzilin A as Improved Antimetastatic Agents","authors":"Noah M. Moriarty, Annaleigh M. Benton, Lauren E. Gartenhaus, Andrew R. Nelson, Haley A. Harper, Carli J. McMahan, Bennett D. Elzey, Jason A. Hanna* and Elizabeth I. Parkinson*, ","doi":"10.1021/acsbiomedchemau.4c0008710.1021/acsbiomedchemau.4c00087","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.4c00087https://doi.org/10.1021/acsbiomedchemau.4c00087","url":null,"abstract":"<p >Vesicle Amine Transport-1 (VAT1) is a protein that is overexpressed in many cancers, including breast cancer, glioblastoma, and angiosarcoma. High VAT1 expression correlates with poor overall survival, and genetic knockout models of VAT1 indicate potent antimigratory activity, suggesting that VAT1 is a promising antimetastasis target. Recently, the natural product neocarzilin A (NCA) from <i>Streptomyces carzinostaticus</i> was reported to be the first validated small-molecule inhibitor of VAT1, having strong activity in metastasis models of angiosarcoma and breast cancer. While knockdown of VAT1 has no effect on cell viability, NCA has significant cytotoxicity, suggesting that NCA is not selective for VAT1. Additionally, NCA has poor aqueous solubility, making <i>in vivo</i> administration of NCA challenging and thus limiting its therapeutic potential. Here, we report the design, synthesis, bioactivity, and pharmacokinetics of novel NCA derivatives with improved drug-like properties. Specifically, we have developed derivatives with altered warheads, replacing chlorines on the trichloroketone with fluorines. Using a modified synthetic route, we accessed NCA derivatives with greater than 25-fold improvements in solubility and 30-fold improvements in the antimigratory to antiproliferative bioactivity ratio. The two best derivatives, along with the parent, were analyzed for oral bioavailability, with the two more soluble derivatives showing greatly improved bioavailability. Overall, these studies have resulted in the development of VAT1 inhibitors with improved properties, which will enable further study of the pharmacological inhibition of VAT1 as an antimetastatic strategy. Additionally, these studies provide insights into novel trihalomethyl ketone warheads and identify chlorodifluoroketone as a potent and selective new warhead.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"4 6","pages":"331–341 331–341"},"PeriodicalIF":3.8,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsbiomedchemau.4c00087","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142843990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}