Discovery of 6-Diazo-5-oxo- l -norleucine (DON) Prodrugs with Enhanced CSF Delivery in Monkeys: A Potential Treatment for Glioblastoma
Abstract
The glutamine antagonist 6-diazo-5-oxo-l-norleucine (DON, 1) has shown robust anticancer efficacy in preclinical and clinical studies, but its development was halted due to marked systemic toxicities. Herein we demonstrate that DON inhibits glutamine metabolism and provides antitumor efficacy in a murine model of glioblastoma, although toxicity was observed. To enhance DON's therapeutic index, we utilized a prodrug strategy to increase its brain delivery and limit systemic exposure. Unexpectedly, simple alkyl ester-based prodrugs were ineffective due to chemical instability cyclizing to form a unique diazo-imine. However, masking both DON's amine and carboxylate functionalities imparted sufficient chemical stability for biological testing. While these dual moiety prodrugs exhibited rapid metabolism in mouse plasma, several provided excellent stability in monkey and human plasma. The most stable compound (5c, methyl-POM-DON-isopropyl-ester) was evaluated in monkeys, where it achieved 10-fold enhanced cerebrospinal fluid to plasma ratio versus DON. This strategy may provide a path to DON utilization in glioblastoma multiforme patients.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
DON was identified as a potential anti-cancer glutamine antagonist more than half a century ago, but its lack of substrate specificity caused too much off-target toxicity.
“6-Diazo-5-oxo-l-norleucine (DON) was identified as a potential anti-cancer glutamine antagonist more than a half century ago201, but its lack of substrate specificity provided too much off-target toxicity202.”
From Pancreatic cancer: Advances and challenges (2023) · cites this paper as [202]
Cites
Radiotherapy Plus Concomitant and Adjuvant Temozolomide for Glioblastoma
Stupp, R., Mason, W. P., van den Bent, M. J., Weller, M., Fisher, B., Taphoorn, M. J. B., Belanger, K., Brandes, A. A., Marosi, C., Bogdahn, U., et al. (2005) · New England Journal of Medicine · doi:10.1056/nejmoa043330 · Cited by 21,807 · see citing works
Stupp, R., Hegi, M. E., Mason, W. P., van den Bent, M. J., TAPHOORN, M. J. B., Janzer, R. C., Ludwin, S. K., Allgeier, A., Fisher, B., Bélanger, K., et al. (2009) · The Lancet Oncology · doi:10.1016/s1470-2045(09)70025-7 · Cited by 7,667 · see citing works
Ostrom, Q. T., Gittleman, H. R., Fulop, J., Liu, M. B., Blanda, R., Kromer, C. M., Wolinsky, Y., Kruchko, C. A., & Barnholtz‐Sloan, J. S. (2015) · Neuro-Oncology · doi:10.1093/neuonc/nov189 · Cited by 2,142 · see citing works
Glutamine addiction: a new therapeutic target in cancer
Wise, D. R., & Thompson, C. B. (2010) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2010.05.003 · Cited by 1,770 · see citing works
Prodrugs: design and clinical applications
Rautio, J., Kumpulainen, H., Heimbach, T., Oliyai, R., Oh, D.-M., Järvinen, T., & Savolainen, J. (2008) · Nature Reviews Drug Discovery · doi:10.1038/nrd2468 · Cited by 1,645 · see citing works
Glutamine and cancer: cell biology, physiology, and clinical opportunities
Hensley, C. T., Wasti, A. T., & DeBerardinis, R. J. (2013) · Journal of Clinical Investigation · doi:10.1172/jci69600 · Cited by 1,262 · see citing works
Glucose-Independent Glutamine Metabolism via TCA Cycling for Proliferation and Survival in B Cells
Le, A., Lane, A. N., Hamaker, M., Bose, S., Gouw, A., Barbi, J., Tsukamoto, T., Rojas, C. J., Slusher, B. S., Zhang, H., et al. (2012) · Cell Metabolism · doi:10.1016/j.cmet.2011.12.009 · Cited by 1,095 · see citing works
Self-citationAntitumor activity of the glutaminase inhibitor CB-839 in triple-negative breast cancer
Gross, M. I., Demo, S. D., Dennison, J. B., Chen, L., Chernov-Rogan, T., Goyal, B., Janes, J. R., Laidig, G. J., Lewis, E. R., Li, J., et al. (2014) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-13-0870 · Cited by 1,068 · see citing works
How cancer metabolism is tuned for proliferation and vulnerable to disruption
Schulze, A., & Harris, A. L. (2012) · Nature · doi:10.1038/nature11706 · Cited by 927 · see citing works
Uptake and Metabolism of Plasma Glutamine by the Small Intestine
Windmueller, H. G., & Spaeth, A. E. (1974) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)42329-6 · Cited by 791 · see citing works
EANO guideline for the diagnosis and treatment of anaplastic gliomas and glioblastoma
Weller, M., van den Bent, M. J., Hopkins, K., Tonn, J. C., Stupp, R., Falini, A., Moyal, E. C., Frappaz, D., Henriksson, R., Balañá, C., et al. (2014) · The Lancet Oncology · doi:10.1016/s1470-2045(14)70011-7 · Cited by 754 · see citing works
Drug Delivery Systems, CNS Protection, and the Blood Brain Barrier
Upadhyay, R. K. (2014) · BioMed Research International · doi:10.1155/2014/869269 · Cited by 532 · see citing works
Enzymes involved in the bioconversion of ester-based prodrugs
Liederer, B. M., & Borchardt, R. T. (2006) · Journal of Pharmaceutical Sciences · doi:10.1002/jps.20542 · Cited by 378 · see citing works
Compensatory glutamine metabolism promotes glioblastoma resistance to mTOR inhibitor treatment
Tanaka, K., Sasayama, T., Irino, Y., Takata, K., Nagashima, H., Satoh, N., Kyotani, K., Mizowaki, T., Imahori, T., Ejima, Y., et al. (2015) · Journal of Clinical Investigation · doi:10.1172/jci78239 · Cited by 243 · see citing works
Metabolism and action of amino acid analog anti-cancer agents
Ahluwalia, G. S., Grem, J. L., Hao, Z., & Cooney, D. A. (1990) · Pharmacology & Therapeutics · doi:10.1016/0163-7258(90)90094-i · Cited by 241 · see citing works
Genetic Alterations and Oncogenic Pathways Associated with Breast Cancer Subtypes
Hu, X., Stern, H. M., Ge, L., O'Brien, C. L., Haydu, L. E., Honchell, C. D., Haverty, P. M., Peters, B. A., Wu, T. D., Amler, L. C., et al. (2009) · Molecular Cancer Research · doi:10.1158/1541-7786.mcr-08-0107 · Cited by 218 · see citing works
Shukla, K. H., Ferraris, D. V., Thomas, A. G., Stathis, M. J., Duvall, B. R., Delahanty, G., Alt, J., Rais, R., Rojas, C. J., Gao, P., et al. (2012) · Journal of Medicinal Chemistry · doi:10.1021/jm301191p · Cited by 199 · see citing works
Self-citationEshleman, J. S., Carlson, B. L., Mladek, A. C., Kastner, B. D., Shide, K. L., & Sarkaria, J. N. (2002) · PubMed · Cited by 197 · see citing works
Glutaminase: A Hot Spot For Regulation Of Cancer Cell Metabolism?
Erickson, J. W., & Cerione, R. A. (2010) · Oncotarget · doi:10.18632/oncotarget.208 · Cited by 155 · see citing works
Cervantes-Madrid, D., Romero, Y., & Dueñas‐González, A. (2015) · BioMed Research International · doi:10.1155/2015/690492 · Cited by 119 · see citing works
Trypanosoma brucei CTP synthetase: A target for the treatment of African sleeping sickness
Hofer, A., Steverding, D., Chabes, A., et al. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.111139498 · Cited by 116 · see citing works
Structural Basis for the Active Site Inhibition Mechanism of Human Kidney-Type Glutaminase (KGA)
Thangavelu, K., Chong, Q. Y., Low, B. C., & Sivaraman, J. (2014) · Scientific Reports · doi:10.1038/srep03827 · Cited by 108 · see citing works
Tougou, K., Nakamura, A., Watanabe, S., et al. (1998) · PubMed · Cited by 100 · see citing works
6-Diazo-5-oxo-L-norleucine, A New Tumor-inhibitory Substance. II.1 Isolation and Characterization2
Dion, H. W., Fusari, S. A., Jakubowski, Z. L., et al. (1956) · Journal of the American Chemical Society · doi:10.1021/ja01594a036 · Cited by 96 · see citing works
Fortner, J. G. (1957) · Cancer · doi:10.1002/1097-0142(195711/12)10:6 · Cited by 76 · see citing works
Lee, Y., Yang, C., Chang, H.-Y., Hsu, H., Chen, I.-S., Chang, H.-S., Lee, C., Lee, J., Kumar, C. R., Qiu, Y., et al. (2014) · Oncotarget · doi:10.18632/oncotarget.2173 · Cited by 73 · see citing works
Earhart, R. H., Amato, D. J., Chang, A. Y.-C., Borden, E. C., Shiraki, M., Dowd, M. E., Comis, R. L., Davis, T. E., & Smith, T. J. (1990) · Investigational New Drugs · doi:10.1007/bf00216936 · Cited by 71 · see citing works
Tumor Metabolism of Malignant Gliomas
Ru, P., Williams, T. M., Chakravarti, A., & Guo, D. (2013) · Cancers · doi:10.3390/cancers5041469 · Cited by 71 · see citing works
Design and evaluation of novel glutaminase inhibitors
McDermott, L. A., Iyer, P. C., Vernetti, L., Rimer, S., Sun, J., Boby, M. L., Yang, T., Fioravanti, M., O’Neill, J., Wang, L., et al. (2016) · Bioorganic & Medicinal Chemistry · doi:10.1016/j.bmc.2016.03.009 · Cited by 68 · see citing works
Species-dependent and site-specific intestinal metabolism of ester prodrugs
van Gelder, J., Shafiee, M., Clercq, E. D., PENNINCKX, F. M., den Mooter, G. V., Kinget, R., & Augustijns, P. (2000) · International Journal of Pharmaceutics · doi:10.1016/s0378-5173(00)00507-x · Cited by 59 · see citing works
Harding, J. J., Telli, M. L., Münster, P. N., Le, M. H., Molineaux, C. J., Bennett, M. K., Mittra, E., Burris, H. A., Clark, A. S., Dunphy, M. P., et al. (2015) · Journal of Clinical Oncology · doi:10.1200/jco.2015.33.15_suppl.2512 · Cited by 58 · see citing works
Lynch, G., Kemeny, N., & Casper, E. (1982) · American Journal of Clinical Oncology · doi:10.1097/00000421-198210000-00014 · Cited by 53 · see citing works
Phase I study and clinical pharmacology of 6-diazo-5-oxo-L-norleucine (DON)
Rahman, A., Smith, F. P., Luc, P.-V. T., & Woolley, P. V. (1985) · Investigational New Drugs · doi:10.1007/bf00170760 · Cited by 49 · see citing works
Influence of glutamine on the growth of human glioma and medulloblastoma in culture.
Dranoff, G., Elion, G. B., Friedman, H. S., Campbell, G. L., & Bigner, D. D. (1985) · PubMed · Cited by 49 · see citing works
Mitochondrial p32 is upregulated in Myc expressing brain cancers and mediates glutamine addiction
Fogal, V., Babić, I., Chao, Y. S., Pastorino, S., Mukthavaram, R., Jiang, P., Cho, Y., Pingle, S. C., Crawford, J. R., Piccioni, D. E., et al. (2014) · Oncotarget · doi:10.18632/oncotarget.2708 · Cited by 45 · see citing works
Pharmacokinetic and phase I study of intravenous DON (6-diazo-5-oxo-L-norleucine) in children
Sullivan, M. P., Nelson, J. A., Feldman, S. L., et al. (1988) · Cancer Chemotherapy and Pharmacology · doi:10.1007/bf00262746 · Cited by 40 · see citing works
SUPPRESSION OF URIC ACID SYNTHESIS IN THE GOUTY HUMAN BY THE USE OF 6-DIAZO-5-OXO-L-NORLEUCINE*
Grayzel, A. I., Seegmiller, J. E., & Love, E. (1960) · Journal of Clinical Investigation · doi:10.1172/jci104057 · Cited by 39 · see citing works
Potter, M. C., Baxter, V. K., Mathey, R. W., Alt, J., Rojas, C., Griffin, D. E., & Slusher, B. S. (2015) · Journal of NeuroVirology · doi:10.1007/s13365-015-0314-6 · Cited by 32 · see citing works
Self-citationWillis, R. C., & Seegmiller, J. E. (1977) · Journal of Cellular Physiology · doi:10.1002/jcp.1040930308 · Cited by 32 · see citing works
Dranoff, G., Elion, G. B., Friedman, H. S., et al. (1985) · PubMed · Cited by 25 · see citing works
Interspecies Differences in the Metabolism of a Multiester Prodrug by Carboxylesterases
Fu, J., Pacyniak, E., Leed, M. G. D., Sadgrove, M. P., Marson, L., & Jay, M. (2015) · Journal of Pharmaceutical Sciences · doi:10.1002/jps.24632 · Cited by 23 · see citing works
Pharmacokinetics of dapsone and amino acid prodrugs of dapsone.
Pochopin, N. L., Charman, W. N., & Stella, V. J. (1994) · Drug Metabolism and Disposition · doi:10.1016/s0090-9556(25)07567-1 · Cited by 20 · see citing works
Barclay, R. K., & Phillipps, M. A. (1966) · PubMed · Cited by 20 · see citing works
Alt, J., Potter, M. C., Rojas, C. J., & Slusher, B. S. (2015) · Analytical Biochemistry · doi:10.1016/j.ab.2015.01.001 · Cited by 18 · see citing works
Self-citationSakamoto, F., IKEDA, S., Kondo, H., et al. (1985) · Chemical and Pharmaceutical Bulletin · doi:10.1248/cpb.33.4870 · Cited by 18 · see citing works
Alterations in Calcium Metabolism in Cancer Patients Treated with 6-Diazo-5-Oxo-L-Norleucine.
Myers, W. P. L., & Magill, G. B. (1956) · Experimental Biology and Medicine · doi:10.3181/00379727-93-22741 · Cited by 13 · see citing works
COUTTS, I. G. C., & Saint, R. E. (1998) · Tetrahedron Letters · doi:10.1016/s0040-4039(98)00402-x · Cited by 13 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Pancreatic cancer: Advances and challenges
Halbrook, C. J., Lyssiotis, C. A., Pasca di Magliano, M., & Maitra, A. (2023) · Cell · doi:10.1016/j.cell.2023.02.014 · Cited by 1,321 · see citing works
Glutamine blockade induces divergent metabolic programs to overcome tumor immune evasion
Leone, R. D., Zhao, L., Englert, J. M., Sun, I.-M., Oh, M.-H., Sun, I.-H., Arwood, M. L., Bettencourt, I. A., Patel, C. H., Wen, J., et al. (2019) · Science · doi:10.1126/science.aav2588 · Cited by 1,169 · see citing works
Self-citationOh, M.-H., Sun, I.-H., Zhao, L., Leone, R. D., Sun, I.-M., Xu, W., Collins, S. L., Tam, A. J., Blosser, R. L., Patel, C. H., et al. (2020) · Journal of Clinical Investigation · doi:10.1172/jci131859 · Cited by 456 · see citing works
Self-citationTargeting tumor-intrinsic hexosamine biosynthesis sensitizes pancreatic cancer to anti-PD1 therapy
Sharma, N. S., Gupta, V. K., Garrido, V. T., Hadad, R., Durden, B. C., Kesh, K., Giri, B., Ferrantella, A., Dudeja, V., Saluja, A., et al. (2019) · Journal of Clinical Investigation · doi:10.1172/jci127515 · Cited by 203 · see citing works
- Authors
- Rais, R., Jančařík, A., Tenora, L., Nedelcovych, M., Alt, J., Englert, J., Rojas, C., Le, A., Elgogary, A., Tan, J., Monincová, L., Pate, K., Adams, R., Ferraris, D., Powell, J., Majer, P., & Slusher, B. S.
- Affiliations
- Johns Hopkins University (US)
- Czech Academy of Sciences (CZ)
- McDaniel College (US)
- Corresponding author
- Majer, Pavel
- Slusher, Barbara S.
- ORCID
- Venue
- Journal of Medicinal Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-09-06
- Volume
- 59
- Issue
- 18
- Pages
- 8621-8633
- ISSN
- 0022-2623 (ISSN-L)
- 1520-4804
- OpenAlex
- W2516129687
- Cited by
- 138
- References
- 47
- Citations by year
- 2026: 11 · 2025: 16 · 2024: 20 · 2023: 22 · 2022: 14 · 2021: 18 · 2020: 7 · 2019: 14 · 2018: 11 · 2017: 5
- FWCI
- 7.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Medicinal Chemistry (publishedVersion)
- PubMed (publishedVersion)
- ASEP (submittedVersion, public-domain)
- Funders
- Ústav organické chemie a biochemie AV CR, v.v.i. (RVO 61388963)
- Maryland Technology Development Corporation
- Topics
- Health Sciences › Medicine › Genetics › Glioma Diagnosis and Treatment
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis and biological activity
- Keywords
- 6-diazo-5-oxo-L-norleucine, prodrugs, glioblastoma, brain drug delivery, cerebrospinal fluid, plasma stability, antitumor efficacy, systemic toxicity
- MeSH terms
- Animals, Haplorhini, Antimetabolites, Antineoplastic, Brain Neoplasms, Diazooxonorleucine, Female, Glioblastoma, Glutamine, Humans, Mice, Nude, Prodrugs, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review