26 results on '"Patterson, Adam"'
Search Results
2. Implementing an Acute Text-Based Wellness Alert System for Pediatric Residents: A Pilot Study.
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Hart, Rebecca J. and Patterson, Adam
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PILOT projects ,HOSPITAL medical staff ,FISHER exact test ,PSYCHOSOCIAL factors ,PEDIATRICIANS ,HEALTH ,INSTANT messaging ,CHI-squared test ,DESCRIPTIVE statistics ,OCCUPATIONAL health services - Abstract
The article focuses on addressing the well-being of pediatric residents through the implementation and evaluation of a text-message-based wellness "check-in" system, specifically targeting acute stressors. The topics include the feasibility and satisfaction of residents with the implemented program, which involved real-time responses to acute distressing events, demonstrating its potential to improve awareness and support for residents' well-being.
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- 2023
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3. Changes in blood pressure and heart rate during sedation with ketamine in the pediatric ED.
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Patterson, Adam C., Wadia, Shernaz A., Lorenz, Douglas J., and Stevenson, Michelle D.
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Background: Ketamine is commonly used in the emergency department for short, painful procedures. We describe changes in blood pressure (BP) and heart rate (HR) during procedural sedation with ketamine, as these changes have not been well described in children.Methods: We performed a secondary analysis of a prospective, observational study involving children aged 8 to 18 years who received procedural sedation with ketamine in a pediatric emergency department. Serial vital signs and sedation scores were recorded from baseline until recovery from ketamine procedural sedation. Time of orthopedic manipulation was also recorded. Linear mixed-effect models were used to evaluate changes in systolic BP (SBP), diastolic BP (DBP), and HR using 3 sedation strata: presedation (baseline), sedated (ketamine administered and patient deeply sedated), and recovery (ketamine administered with patient minimally sedated), controlling for age and weight.Results: Sixty children were enrolled; 10 were excluded due to missing manipulation time. A total of 394 observations were recorded. Mean sedated SBP, DBP, and HR were 8 mm Hg, 4 mm Hg, and 13 beats/min higher than presedation SBP (P<.001), DBP (P<.01), and HR (P<.001), respectively. Mean sedated SBP and DBP were 3 and 4 mm Hg higher than SBP (P=.006) and DBP (P<.01) during recovery. Manipulation increased mean SBP by 5 mm Hg (P<.001), mean DBP by 7 mm Hg (P<.001), and mean HR by 1 beat/min (P=.35).Conclusions: Ketamine administered during procedural sedation for painful procedures causes a statistically significant but modest increase in SBP, DBP, and HR. Orthopedic manipulation further increases BP. [ABSTRACT FROM AUTHOR]- Published
- 2017
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4. Synthesis and cytotoxicity of pyranonaphthoquinone natural product analogues under bioreductive conditions.
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Heapy, Amanda M., Patterson, Adam V., Smaill, Jeff B., Jamieson, Stephen M.F., Guise, Christopher P., Sperry, Jonathan, Hume, Paul A., Rathwell, Kris, and Brimble, Margaret A.
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NAPHTHOQUINONE , *NATURAL products , *CELL-mediated cytotoxicity , *CHEMICAL synthesis , *MOIETIES (Chemistry) , *BREAST cancer , *CELL lines , *NICOTINAMIDE adenine dinucleotide phosphate - Abstract
Abstract: We have synthesised a focused library of derivatives of natural products containing the pyranonaphthoquinone moiety including the first report of such a scaffold with an appended tetrazole functionality. Examples include kalafungin derivatives as well as analogues of nanaomycin and eleutherin. These compounds were assessed for cytotoxic activation by breast cancer cell lines engineered to express the prototypic human one- and two-electron quinone bioreductive enzymes, NADPH: cytochrome P450 oxidoreductase (POR) and NAD(P)H: quinoneoxidoreductase 1 (NQO1; DT-diaphorase), respectively. Several compounds were observed to be cytotoxic at sub-micromolar level and a pattern of increased aerobic potency was observed in cells over expressing POR. A subset of analogues was assessed under anoxic conditions, where cytotoxicity was reduced, implicating redox cycling as a major mechanism of toxicity. The substrate specificity for reductive enzymes is relevant to the future design of bioreductive prodrugs to treat cancer. [Copyright &y& Elsevier]
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- 2013
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5. Oxygen Dependence and Extravascular Transport of Hypoxia-Activated Prodrugs: Comparison of the Dinitrobenzamide Mustard PR-104A and Tirapazamine
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Hicks, Kevin O., Myint, Hilary, Patterson, Adam V., Pruijn, Frederik B., Siim, Bronwyn G., Patel, Kashyap, and Wilson, William. R.
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HYPOXEMIA , *ANTIBODY-directed enzyme prodrug therapy , *PHARMACOKINETICS , *MEDICAL radiology - Abstract
Purpose: To compare oxygen dependence and tissue transport properties of a new hypoxia-activated prodrug, PR-104A, with tirapazamine, and to evaluate the implications for antitumor activity when combined with radiotherapy. Methods and Materials: Oxygen dependence of cytotoxicity was measured by clonogenic assay in SiHa cell suspensions. Tissue transport parameters were determined using SiHa multicellular layers. Spatially resolved pharmacokinetic (PK) and pharmacodynamic (PD) models were developed to predict cell killing in SiHa tumors and tested by clonogenic assay 18 h after treatment with the corresponding phosphate ester, PR-104. Results: The K-value (oxygen concentration to halve cytotoxic potency) of PR-104A was 0.126 ± 0.021 μM (10-fold lower than tirapazamine at 1.30 ± 0.28 μM). The diffusion coefficient of PR-104A in multicellular layers (4.42 ± 0.15 × 10−7 cm2 s−1) was lower than that of tirapazamine (1.30 ± 0.05 × 10−6 cm2 s−1) but PK modeling predicted better penetration to hypoxic cells in tumors because of its slower metabolism. The tirapazamine PK/PD model successfully predicted the measured activity in combination with single-dose radiation against SiHa tumors, and the PR-104A model underpredicted the activity, which was greater for PR-104 than for tirapazamine (at equivalent host toxicity) both with radiation and as a single agent. Conclusion: PR-104/PR-104A has different PK/PD properties from tirapazamine and superior activity with single-dose radiotherapy against SiHa xenografts. We have inferred that PR-104A is better able to kill cells at intermediate partial pressure of oxygen in tumors than implied by the PK/PD model, most likely because of a bystander effect resulting from diffusion of its activated metabolites from severely hypoxic zones. [Copyright &y& Elsevier]
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- 2007
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6. Radiolytic and cellular reduction of a novel hypoxia-activated cobalt(III) prodrug of a chloromethylbenzindoline DNA minor groove alkylator
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Ahn, G-One, Botting, K. Jane, Patterson, Adam V., Ware, David C., Tercel, Moana, and Wilson, William R.
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PRODRUGS , *COBALT , *TUMORS , *BIOLOGICAL assay - Abstract
Abstract: Metabolic reduction can be used to activate prodrugs in hypoxic regions of tumours, but reduction by ionising radiation is also theoretically attractive. Previously, we showed that a cobalt(III) complex containing 8-hydroxyquinoline (8-HQ) and cyclen ligands releases 8-HQ efficiently on irradiation in hypoxic solutions [Ahn G-O, Ware DC, Denny WA, Wilson WR. Optimization of the auxiliary ligand shell of cobalt(III)(8-hydroxyquinoline) complexes as model hypoxia-selective radiation-activated prodrugs. Radiat Res 2004;162:315–25]. Here we investigate an analogous Co(III) complex containing the potent DNA minor groove alkylator azachloromethylbenzindoline (azaCBI, 1 ) to determine whether it releases 1 on radiolytic and/or enzymatic reduction under hypoxia. Monitoring by HPLC, the azaCBI ligand in the Co(III)(cyclen)(azaCBI) complex ( 2 ) slowly hydrolysed in aqueous solution, in contrast to the free ligand 1 which readily converted to its reactive cyclopropyl form. Irradiation of 2 (30–50μM) in hypoxic solutions released 1 with yields of 0.57μmol/J in formate buffer and 0.13μmol/J in human plasma. Using bioassay methods, cytotoxic activation by irradiation of 2 at 1μM in hypoxic plasma was readily detectable at clinically relevant doses (≥1Gy), with a estimated yield of 1 of 0.075μmol/J. Release of 1 from 2 was also observed in hypoxic HT29 cultures without radiation, with subsequent conversion of 1 to its O-glucuronide. Surprisingly, overexpression of human cytochrome P450 reductase in A549 cells did not increase the rate of metabolic reduction of 2 , suggesting that other reductases and/or non-enzymatic reductants are responsible. Thus the cobalt(III) complex 2 is a promising prodrug capable of being activated to release a very potent cytotoxin when reduced by either ionising radiation or cells under hypoxic conditions. [Copyright &y& Elsevier]
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- 2006
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7. Synthesis and antiproliferative activity of culicinin D analogues containing simplified AHMOD-based residues.
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Stubbing, Louise A., Kavianinia, Iman, Abbattista, Maria R., Harris, Paul W.R., Smaill, Jeff B., Patterson, Adam V., and Brimble, Margaret A.
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AMINO acids , *STRUCTURE-activity relationships - Abstract
Culicinin D is a 10 amino acid peptaibol containing a rare and synthetically challenging (2 S ,4 S ,6 R)-AHMOD residue, that exhibits potent antiproliferative activity against MDA-MB-468 cells. An SAR study focusing on replacement of the AHMOD residue was undertaken, culminating in the revelation that a 6-hydroxy or 6-keto substituent was essential to retain potent low nanomolar antiproliferative activity. Image 1 [ABSTRACT FROM AUTHOR]
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- 2019
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8. Prototyping kinase inhibitor-cytotoxin anticancer mutual prodrugs activated by tumour hypoxia: A chemical proof of concept study.
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Sansom, Geraud N., Kirk, Nicholas S., Guise, Christopher P., Anderson, Robert F., Smaill, Jeff B., Patterson, Adam V., and Kelso, Michael J.
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PRODRUGS , *PROOF of concept , *FRAGMENTATION reactions , *HYPOXEMIA , *CHEMICAL reduction , *REDUCTION potential - Abstract
Graphical abstract Abstract Amide- and ester-linked kinase inhibitor-cytotoxin conjugates were rationally designed and synthesised as prototype hypoxia-activated anticancer mutual prodrugs. Chemical reduction of an aryl nitro trigger moiety was shown to initiate a spontaneous cyclisation/fragmentation reaction that simultaneously released the kinase inhibitor semaxanib (SU5416) and the amine- or alcohol-linked cytotoxin from the prodrugs. Preliminary cell testing and reduction potential measurements support optimisation of the compounds towards tumour-selective mutual prodrugs. [ABSTRACT FROM AUTHOR]
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- 2019
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9. Evaluating the abilities of diverse nitroaromatic prodrug metabolites to exit a model Gram negative vector for bacterial-directed enzyme-prodrug therapy.
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Chan-Hyams, Jasmine V.E., Copp, Janine N., Smaill, Jeff B., Patterson, Adam V., and Ackerley, David F.
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NITROAROMATIC compounds , *DRUG metabolism , *PRODRUGS , *GRAM-negative bacteria , *DISEASE vectors , *CELL culture - Abstract
Graphical abstract Abstract Gene-directed enzyme-prodrug therapy (GDEPT) employs tumour-tropic vectors including viruses and bacteria to deliver a genetically-encoded prodrug-converting enzyme to the tumour environment, thereby sensitising the tumour to the prodrug. Nitroreductases, able to activate a range of promising nitroaromatic prodrugs to genotoxic metabolites, are of great interest for GDEPT. The bystander effect (cell-to-cell transfer of activated prodrug metabolites) has been quantified for some nitroaromatic prodrugs in mixed multilayer human cell cultures, however while these provide a good model for viral DEPT (VDEPT) they do not inform on the ability of these prodrug metabolites to exit bacterial vectors (relevant to bacterial-DEPT (BDEPT)). To investigate this we grew two Escherichia coli strains in co-culture; an activator strain expressing the nitroreductase E. coli NfsA and a recipient strain containing an SOS-GFP DNA damage responsive gene construct. In this system, induction of GFP by reduced prodrug metabolites can only occur following their transfer from the activator to the recipient cells. We used this to investigate five clinically relevant prodrugs: metronidazole, CB1954, nitro-CBI-DEI, and two dinitrobenzamide mustard prodrug analogues, PR-104A and SN27686. Consistent with the bystander efficiencies previously measured in human cell multilayers, reduced metronidazole exhibited little bacterial cell-to-cell transfer, whereas nitro-CBI-DEI was passed very efficiently from activator to recipient cells post-reduction. However, in contrast with observations in human cell multilayers, the nitrogen mustard prodrug metabolites were not effectively passed between the two bacterial strains, whereas reduced CB1954 was transferred efficiently. Using nitroreductase enzymes that exhibit different biases for the 2- versus 4-nitro substituents of CB1954, we further showed that the 2-nitro reduction products exhibit substantially higher levels of bacterial cell-to-cell transfer than the 4-nitro reduction products, consistent with their relative bystander efficiencies in human cell culture. Overall, our data suggest that prodrugs may differ in their suitability for VDEPT versus BDEPT applications and emphasise the importance of evaluating an enzyme-prodrug partnership in an appropriate context for the intended vector. [ABSTRACT FROM AUTHOR]
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- 2018
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10. Discovery and optimization of 1-(1H-indol-1-yl)ethanone derivatives as CBP/EP300 bromodomain inhibitors for the treatment of castration-resistant prostate cancer.
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Xiang, Qiuping, Wang, Chao, Zhang, Yan, Xue, Xiaoqian, Song, Ming, Zhang, Cheng, Li, Chenchang, Wu, Chun, Li, Kuai, Hui, Xiaoyan, Zhou, Yulai, Smaill, Jeff B., Patterson, Adam V., Wu, Donghai, Ding, Ke, and Xu, Yong
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CREB protein , *CASTRATION-resistant prostate cancer , *PROTEIN expression , *PROTEIN domains , *MESSENGER RNA , *CANCER treatment - Abstract
The CREB (cAMP responsive element binding protein) binding protein (CBP) and its homolog EP300 have emerged as new therapeutic targets for the treatment of cancer and inflammatory diseases. Here we report the identification, optimization and evaluation of 1-(1 H -indol-1-yl)ethanone derivatives as CBP/EP300 inhibitors starting from fragment-based virtual screening (FBVS). A cocrystal structure of the inhibitor ( 22e ) in complex with CBP provides a solid structural basis for further optimization. The most potent compound 32h binds to the CBP bromodomain and has an IC 50 value of 0.037 μM in the AlphaScreen assay which was 2 times more potent than the reported CBP bromodomain inhibitor SGC-CBP30 in our hands. 32h also exhibit high selectivity for CBP/EP300 over other bromodomain-containing proteins. Notably, the ester derivative ( 29h ) of compound 32h markedly inhibits cell growth in several prostate cancer cell lines including LNCaP, 22Rv1 and LNCaP derived C4-2B. Compound 29h suppresses the mRNA expression of full length AR (AR-FL), AR target genes and other oncogene in LNCaP cells. 29h also reduces the expression of PSA, the biomarker of prostate cancer. CBP/EP300 inhibitor 29h represents a promising lead compound for the development of new therapeutics for the treatment of castration-resistant prostate cancer. [ABSTRACT FROM AUTHOR]
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- 2018
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11. 2-Oxo-3, 4-dihydropyrimido[4, 5-d]pyrimidinyl derivatives as new irreversible pan fibroblast growth factor receptor (FGFR) inhibitors.
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Li, Xueqiang, Guise, Christopher P., Taghipouran, Rana, Yosaatmadja, Yuliana, Ashoorzadeh, Amir, Paik, Woo-Kyong, Squire, Christopher J., Jiang, Shuang, Luo, Jinfeng, Xu, Yong, Tu, Zheng-Chao, Lu, Xiaoyun, Ren, Xiaomei, Patterson, Adam V., Smaill, Jeff B., and Ding, Ke
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PYRIMIDINE derivatives , *FIBROBLAST growth factor receptors , *ANTINEOPLASTIC agents , *DRUG design , *ENZYME inhibitors - Abstract
A series of 2-oxo-3, 4-dihydropyrimido[4,5- d ]-pyrimidinyl derivatives were designed and synthesized as new irreversible inhibitors of the FGFR family. One of the most promising compounds 2l potently inhibited FGFR1/2/3 with IC 50 values of 1.06, 0.84 and 5.38 nM, respectively, whereas its potency against FGFR4 was diminished by an order of magnitude. Compound 2l strongly suppresses the proliferation of FGFR1-amplified H520 non-small cell lung cancer cells, FGFR2-amplified SUM52 breast cancer cells and FGFR3-amplified SW780 bladder cancer cells with low nanomolar IC 50 values, but was significantly less potent against four FGFR-negative cancer cell lines, with low micromolar IC 50 values. Biological investigation also confirmed the irreversible binding of the molecule with the FGFR1-3 target kinases. Compound 2l may serve as a promising new lead for further anticancer drug discovery. [ABSTRACT FROM AUTHOR]
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- 2017
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12. Rational design of an AKR1C3-resistant analog of PR-104 for enzyme-prodrug therapy.
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Mowday, Alexandra M., Ashoorzadeh, Amir, Williams, Elsie M., Copp, Janine N., Silva, Shevan, Bull, Matthew R., Abbattista, Maria R., Anderson, Robert F., Flanagan, Jack U., Guise, Christopher P., Ackerley, David F., Smaill, Jeff B., and Patterson, Adam V.
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ALDO-keto reductases , *ANTINEOPLASTIC agents , *PRODRUGS , *NITROREDUCTASES , *OXIDOREDUCTASES , *CYTOCHROME P-450 , *DRUG design , *THERAPEUTICS - Abstract
The clinical stage anti-cancer agent PR-104 has potential utility as a cytotoxic prodrug for exogenous bacterial nitroreductases expressed from replicating vector platforms. However substrate selectivity is compromised due to metabolism by the human one- and two-electron oxidoreductases cytochrome P450 oxidoreductase (POR) and aldo-keto reductase 1C3 (AKR1C3). Using rational drug design we developed a novel mono-nitro analog of PR-104A that is essentially free of this off-target activity in vitro and in vivo . Unlike PR-104A, there was no biologically relevant cytotoxicity in cells engineered to express AKR1C3 or POR, under aerobic or anoxic conditions, respectively. We screened this inert prodrug analog, SN34507, against a type I bacterial nitroreductase library and identified E. coli NfsA as an efficient bioactivator using a DNA damage response assay and recombinant enzyme kinetics. Expression of E. coli NfsA in human colorectal cancer cells led to selective cytotoxicity to SN34507 that was associated with cell cycle arrest and generated a robust ‘bystander effect’ at tissue-like cell densities when only 3% of cells were NfsA positive. Anti-tumor activity of SN35539, the phosphate pre-prodrug of SN34507, was established in ‘mixed’ tumors harboring a minority of NfsA-positive cells and demonstrated marked tumor control following heterogeneous suicide gene expression. These experiments demonstrate that off-target metabolism of PR-104 can be avoided and identify the suicide gene/prodrug partnership of E. coli NfsA/SN35539 as a promising combination for development in armed vectors. [ABSTRACT FROM AUTHOR]
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- 2016
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13. 2,4-Diarylamino-pyrimidines as kinase inhibitors co-targeting IGF1R and EGFRL858R/T790M.
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Chan, Shingpan, Han, Kun, Qu, Rong, Tong, Linjiang, Li, Yingjun, Zhang, Zhang, Cheng, Huimin, Lu, Xiaoyun, Patterson, Adam, Smaill, Jeff, Ren, Xiaomei, Ding, Jian, Xie, Hua, and Ding, Ke
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PYRIMIDINES , *KINASE inhibitors , *SOMATOMEDIN C , *CANCER cells , *ANTINEOPLASTIC agents - Abstract
IGF1R amplification was recently implied to be related to the secondary acquired resistance against the 2nd or 3rd generation EGFR inhibitor therapies. We have successfully identified a series of 2,4-diarylamino-pyrimidines as new IGF1R/EGFR L858R/T790M co-targeting agents. One of the most promising compounds 8g potently inhibits both kinases with low nanomolar IC 50 values, but is significantly less potent in inhibiting the wild type EGFR. The compound also displays a good kinase selectivity profile against a panel of 468 kinases. Moreover, 8g strongly suppresses the proliferation of CO-1686-resistant H1975-IGF1R cancer cells, suggesting its promising potential as a new lead compound for future anticancer drug discovery. [ABSTRACT FROM AUTHOR]
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- 2015
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14. Identification of one-electron reductases that activate both the hypoxia prodrug SN30000 and diagnostic probe EF5.
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Wang, Jingli, Guise, Chris P., Dachs, Gabi U., Phung, Yen, Hsu, Annie (Huai-Ling), Lambie, Neil K., Patterson, Adam V., and Wilson, William R.
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HYPOXEMIA , *PRODRUGS , *TRIAZINES , *OXIDOREDUCTASES , *CANCER cells , *GENE expression , *IMMUNOSTAINING - Abstract
SN30000 is a second-generation benzotriazine-N-oxide hypoxia-activated prodrug scheduled for clinical trial. Previously we showed that covalent binding of the hypoxia probe EF5 predicts metabolic activation of SN30000 in a panel of cancer cell lines under anoxia, suggesting that they are activated by the same reductases. However the identity of these reductases is unknown. Here, we test whether forced expression of nine oxidoreductases with known or suspected roles in bioreductive prodrug metabolism (AKR1C3, CYB5R3, FDXR, MTRR, NDOR1, NOS2A, NQO1, NQO2 and POR) enhances oxic or anoxic reduction of SN30000 and EF5 by HCT116 cells. Covalent binding of 14C-EF5 and reduction of SN30000 to its 1-oxide and nor-oxide metabolites was highly selective for anoxia in all lines, with significantly elevated anoxic metabolism of both compounds in lines over-expressing POR, MTRR, NOS2A or NDOR1. There was a strong correlation between EF5 binding and SN30000 metabolism under anoxia across the cell lines (R2 = 0.84, p = 0.0001). Antiproliferative potency of SN30000 under anoxia was increased most strongly by overexpression of MTRR and POR. Transcript abundance in human tumours, evaluated using public domain mRNA expression data, was highest for MTRR, followed by POR, NOS2A and NDOR1, with little variation between tumour types. Immunostaining of tissue microarrays demonstrated variable MTRR protein expression across 517 human cancers with most displaying low expression. In conclusion, we have identified four diflavin reductases (POR, MTRR, NOS2A and NDOR1) capable of reducing both SN30000 and EF5, further supporting use of 2-nitroimidazole probes to predict the ability of hypoxic cells to activate SN30000. [ABSTRACT FROM AUTHOR]
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- 2014
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15. A novel fluorometric assay for aldo-keto reductase 1C3 predicts metabolic activation of the nitrogen mustard prodrug PR-104A in human leukaemia cells.
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Jamieson, Stephen M.F., Gu, Yongchuan, Manesh, Donya Moradi, El-Hoss, Jad, Jing, Duohui, MacKenzie, Karen L., Guise, Christopher P., Foehrenbacher, Annika, Pullen, Susan M., Benito, Juliana, Smaill, Jeffrey B., Patterson, Adam V., Mulaw, Medhanie A., Konopleva, Marina, Bohlander, Stefan K., Lock, Richard B., and Wilson, William R.
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FLUORIMETRY , *ALDO-keto reductases , *NITROGEN mustards , *PRODRUGS , *STEROID hormones , *PROSTAGLANDINS - Abstract
Abstract: Aldo-keto reductase 1C3 (AKR1C3, EC 1.1.1.188) metabolises steroid hormones, prostaglandins and xenobiotics, and activates the dinitrobenzamide mustard prodrug PR-104A by reducing it to hydroxylamine PR-104H. Here, we describe a functional assay for AKR1C3 in cells using the fluorogenic probe coumberone (a substrate for all AKR1C isoforms) in conjunction with a specific inhibitor of AKR1C3, the morpholylurea SN34037. We use this assay to evaluate AKR1C3 activity and PR-104A sensitivity in human leukaemia cells. SN34037-sensitive reduction of coumberone to fluorescent coumberol correlated with AKR1C3 protein expression by immunoblotting in a panel of seven diverse human leukaemia cell lines, and with SN34037-sensitive reduction of PR-104A to PR-104H. SN34037 inhibited aerobic cytotoxicity of PR-104A in high-AKR1C3 TF1 erythroleukaemia cells, but not in low-AKR1C3 Nalm6 pre-B cell acute lymphocytic leukaemia (B-ALL) cells, although variation in PR-104H sensitivity confounded the relationship between AKR1C3 activity and PR-104A sensitivity across the cell line panel. AKR1C3 mRNA expression showed wide variation between leukaemia patients, with consistently higher levels in T-ALL than B-ALL. In short term cultures from patient-derived paediatric ALL xenografts, PR-104A was more potent in T-ALL than B-ALL lines, and PR-104A cytotoxicity was significantly inhibited by SN34037 in T-ALL but not B-ALL. Overall, the results demonstrate that SN34037-sensitive coumberone reduction provides a rapid and specific assay for AKR1C3 activity in cells, with potential utility for identifying PR-104A-responsive leukaemias. However, variations in PR-104H sensitivity indicate the need for additional biomarkers for patient stratification. [Copyright &y& Elsevier]
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- 2014
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16. Zinc Finger Nuclease Knock-out of NADPH:Cytochrome P450 Oxidoreductase (POR) in Human Tumor Cell Lines Demonstrates That Hypoxia-activated Prodrugs Differ in POR Dependence.
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Jiechuang Su, Yongchuan Gu, Pruijn, Frederik B., Smaill, Jeff B., Patterson, Adam V., Guise, Christopher P., and Wilson, William R.
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ZINC , *ZINC-finger proteins , *CEREBRAL anoxia , *CELL culture , *CANCER cells - Abstract
Hypoxia, a ubiquitous feature of tumors, can be exploited by hypoxia-activated prodrugs (HAP) that are substrates for oneelectron reduction in the absence of oxygen. NADPH:cytochrome P450 oxidoreductase (POR) is considered one of the major enzymes responsible, based on studies using purified enzyme or forced overexpression in cell lines. To examine the role of POR in HAP activation at endogenous levels of expression, POR knock-outs were generated in HCT116 and SiHa cells by targeted mutation of exon 8 using zinc finger nucleases. Absolute quantitation by proteotypic peptide mass spectrometry of DNA sequence-confirmed multiallelic mutants demonstrated expression of proteins with residual one-electron reductase activity in some clones and identified two (Hko2 from HCT116 and S2ko1 from SiHa) that were functionally null by multiple criteria. Sensitivities of the clones to 11 HAP (six nitroaromatics, three benzotriazine N-oxides, and two quinones) were compared with wild-type and POR-overexpressing cells. All except the quinones were potentiated by POR overexpression. Knocking out POR had a marked effect on antiproliferative activity of the 5-nitroquinoline SN24349 in both genetic backgrounds after anoxic exposure but little or no effect on activity of most other HAP, including the clinical stage 2-nitroimidazole mustard TH-302, dinitrobenzamide mustard PR-104A, and benzotriazine N-oxide SN30000. Clonogenic cell killing and reductive metabolism of PR-104A and SN30000 under anoxia also showed little change in the POR knock-outs. Thus, although POR expression is a potential biomarker of sensitivity to some HAP, identification of other one-electron reductases responsible for HAP activation is needed for their rational clinical development. [ABSTRACT FROM AUTHOR]
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- 2013
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17. Synthesis of substituted 5-bromomethyl-4-nitroimidazoles and use for the preparation of the hypoxia-selective multikinase inhibitor SN29966.
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Lu, Guo-Liang, Ashoorzadeh, Amir, Anderson, Robert F., Patterson, Adam V., and Smaill, Jeff B.
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NITROIMIDAZOLES , *SUBSTITUTION reactions , *CHEMICAL synthesis , *IMIDAZOLES , *HYPOXEMIA , *KINASE inhibitors , *CHEMICAL precursors - Abstract
Abstract: 5-Bromomethyl-4-nitroimidazoles have utility as bioreductive trigger precursors for the preparation of hypoxia-selective prodrugs. Here we describe an efficient two-step synthesis of 5-(bromomethyl)-1-methyl-4-nitro-1H-imidazole, a preferred precursor, employing an N-bromosuccinimide mediated radical bromination. Use of this precursor to prepare SN29966, a promising hypoxia-selective irreversible pan-ErbB inhibitor is reported along with the preparation of four other prodrug candidates. 5-Bromomethyl-4-nitroimidazole analogues bearing electron-donating and electron-withdrawing substituents at the N-1 and C-2 positions are also described. [Copyright &y& Elsevier]
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- 2013
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18. Creation and screening of a multi-family bacterial oxidoreductase library to discover novel nitroreductases that efficiently activate the bioreductive prodrugs CB1954 and PR-104A
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Prosser, Gareth A., Copp, Janine N., Mowday, Alexandra M., Guise, Christopher P., Syddall, Sophie P., Williams, Elsie M., Horvat, Claire N., Swe, Pearl M., Ashoorzadeh, Amir, Denny, William A., Smaill, Jeff B., Patterson, Adam V., and Ackerley, David F.
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OXIDOREDUCTASES , *BACTERIAL enzymes , *NITROREDUCTASES , *PRODRUGS , *DRUG activation , *ANTINEOPLASTIC agents , *DRUG development , *DRUG efficacy - Abstract
Abstract: Two potentially complementary approaches to improve the anti-cancer strategy gene-directed enzyme prodrug therapy (GDEPT) are discovery of more efficient prodrug-activating enzymes, and development of more effective prodrugs. Here we demonstrate the utility of a flexible screening system based on the Escherichia coli SOS response to evaluate novel nitroreductase enzymes and prodrugs in concert. To achieve this, a library of 47 candidate genes representing 11 different oxidoreductase families was created and screened to identify the most efficient activators of two different nitroaromatic prodrugs, CB1954 and PR-104A. The most catalytically efficient nitroreductases were found in the NfsA and NfsB enzyme families, with NfsA homologues generally more active than NfsB. Some members of the AzoR, NemA and MdaB families also exhibited low-level activity with one or both prodrugs. The results of SOS screening in our optimised E. coli reporter strain SOS-R2 were generally predictive of the ability of nitroreductase candidates to sensitise E. coli to CB1954, and of the k cat /K m for each prodrug substrate at a purified protein level. However, we also found that not all nitroreductases express stably in human (HCT-116 colon carcinoma) cells, and that activity at a purified protein level did not necessarily predict activity in stably transfected HCT-116. These results highlight a need for all enzyme-prodrug partners for GDEPT to be assessed in the specific context of the vector and cell line that they are intended to target. Nonetheless, our oxidoreductase library and optimised screens provide valuable tools to identify preferred nitroreductase-prodrug combinations to advance to preclinical evaluation. [Copyright &y& Elsevier]
- Published
- 2013
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19. Identification of human reductases that activate the dinitrobenzamide mustard prodrug PR-104A: A role for NADPH:cytochrome P450 oxidoreductase under hypoxia
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Guise, Chris P., Wang, Anderson T., Theil, Anke, Bridewell, David J., Wilson, William R., and Patterson, Adam V.
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DINITROBENZENES , *NAD(P)H dehydrogenases , *CYTOCHROME P-450 , *PRODRUGS - Abstract
Abstract: Hypoxia is a common trait found in many solid tumours and thus represents a therapeutic target with considerable potential. PR-104, a hypoxia-activated prodrug currently in clinical trial, is a water-soluble phosphate ester which is converted in vivo to the corresponding alcohol, PR-104A. This 3,5-dinitrobenzamide-2-nitrogen mustard is activated by reduction to the corresponding 5-hydroxylamine (PR-104H) and 5-amine (PR-104M) in hypoxic cells. The clinical effectiveness of PR-104 will depend in part on the expression of reductases within tumours that can effect this reduction. Here, we evaluate the roles of NADPH:cytochrome P450 oxidoreductase (CYPOR; E.C.1.6.2.4) and NAD(P)H:quinone oxidoreductase (NQO1; E.C.1.6.99.2) as candidate PR-104A reductases. A weak correlation was observed between NQO1 activity and aerobic cytotoxicity in a panel of eight tumour cell lines. However, overexpression of human NQO1 did not increase cytotoxicity of PR-104A or the formation of PR-104H/M, showing that PR-104A is not a substrate for NQO1. Overexpression of human CYPOR did, however, increase the hypoxic cytotoxicity of PR-104A, and its metabolism to PR-104H and PR-104M, demonstrating it to be a PR-104A reductase. To assess the contribution of CYPOR to overall activation of PR-104A in hypoxic SiHa cells, a combination of siRNA transfection and antisense expression were used to suppress CYPOR protein by 91% (±3%), a phenotype which conferred 45% (±7%) decrease in cytotoxic potency of PR-104A. Regression analysis of all CYPOR depletion data was found to correlate with cytoprotection and metabolism (p <0.001). Residual PR-104A reductase activity could be inhibited by the flavoprotein inhibitor diphenyliodonium. We conclude that CYPOR is an important PR-104A reductase, but that other flavoenzymes also contribute to its activation in hypoxic SiHa cells. [Copyright &y& Elsevier]
- Published
- 2007
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20. Dual responsive promoters to target therapeutic gene expression to radiation-resistant hypoxic tumor cells
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Chadderton, Naomi, Cowen, Rachel L., Sheppard, Freda C.D., Robinson, Suzanne, Greco, Olga, Scott, Simon D., Stratford, Ian J., Patterson, Adam V., and Williams, Kaye J.
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CEREBRAL anoxia , *CANCER cells , *HOSPITAL radiological services , *MEDICAL radiology - Abstract
Purpose: Tumor hypoxia is unequivocally linked to poor radiotherapy outcome. This study aimed to identify enhancer sequences that respond maximally to a combination of radiation and hypoxia for use in genetic radiotherapy approaches.Methods and Materials: The influence of radiation (5 Gy) and hypoxia (1% O2) on reporter-gene expression driven by hypoxia (HRE) and radiation (Egr-1) responsive elements was evaluated in tumor cells grown as monolayers or multicellular spheroids. Hypoxia-inducible factor-1alpha (HIF-1alpha) and HIF-2alpha protein expression was monitored in parallel.Results: Of the sequences tested, an HRE from the phosphoglycerate kinase-1 gene (PGK-18[5+]) was maximally induced in response to hypoxia plus radiation in all 5 cell lines tested. The additional radiation treatment afforded a significant increase in the induction of PGK-18[5+] compared with hypoxia alone in 3 cell lines. HIF-1alpha/2alpha were induced by radiation but combined hypoxia/radiation treatment did not yield a further increase. The dual responsive nature of HREs was maintained when spheroids were irradiated after delivery of HRE constructs in a replication-deficient adenovirus.Conclusions: Hypoxia-responsive enhancer element sequences are dually responsive to combined radiation and hypoxic treatment. Their use in genetic radiotherapy in vivo could maximize expression in the most radio-resistant population at the time of radiation and also exploit microenvironmental changes after radiotherapy to yield additional switch-on. [ABSTRACT FROM AUTHOR]- Published
- 2005
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21. Pharmacological and biological evaluation of a series of substituted 1,4-naphthoquinone bioreductive drugs
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Phillips, Roger M., Jaffar, Mohammed, Maitland, Derek J., Loadman, Paul M., Shnyder, Steven D., Steans, Gillian, Cooper, Patricia A., Race, Amanda, Patterson, Adam V., and Stratford, Ian J.
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NAPHTHOQUINONE , *NAPHTHALENE , *DRUGS , *PHARMACOLOGY - Abstract
The indolequinone compound EO9 has good pharmacodynamic properties in terms of bioreductive activation and selectivity for either NAD(P)H:quinone oxidoreductase-1 (NQO1)-rich aerobic or NQO1-deficient hypoxic cells. However, its pharmacokinetic properties are poor and this fact is believed to be a major reason for EO9''s lack of clinical efficacy. The purpose of this study was to develop quinone-based bioreductive drugs that retained EO9''s good properties, in terms of bioreductive activation, but have improved pharmacokinetic properties. Out of 11 naphthoquinone compounds evaluated, 2-aziridinyl-5-hydroxy-1,4-naphthoquinone (compound 2), 2,3-bis(aziridinyl)-5-hydroxy-1,4-naphthoquinone (compound 3), and 2-aziridinyl-6-hydroxymethyl-1,4-naphthoquinone (compound 11) were selected for further evaluation based on good substrate specificity for NQO1 and selectivity towards NQO1-rich cells in vitro. Compound 3 was of particular interest as it also demonstrated selectivity for NQO1-rich cells under hypoxic conditions. Compound 3 was not metabolised by murine whole blood in vitro (in contrast to compounds 2, 11 and EO9) and pharmacokinetic studies in non-tumour-bearing mice in vivo (at the maximum soluble dose of 60mgkg-1 administered intraperitoneally) demonstrated significant improvements in plasma half-life (16.2min) and AUC values (22.5μMh) compared to EO9 (T1/2 = 1.8min, AUC = 0.184μMh). Compound 3 also demonstrated significant anti-tumour activity against H460 and HCT-116 human tumour xenografts in vivo, whereas EO9 was inactive against these tumours. In conclusion, compound 3 is a promising lead compound that may target both aerobic and hypoxic fractions of NQO1-rich tumours and further studies to elucidate its mechanism of action and improve solubility are warranted. [Copyright &y& Elsevier]
- Published
- 2004
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22. The bioreductive agent RH1 and gamma-irradiation both cause G2/M cell cycle phase arrest and polyploidy in a p53-mutated human breast cancer cell line.
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Kim, Joo-Young, Kim, Chul-Hwan, Stratford, Ian J, Patterson, Adam V, and Hendry, Jolyon H
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HETEROCYCLIC compounds , *BENZOQUINONES , *APOPTOSIS , *BREAST tumors , *CELL cycle , *CELL lines , *CELL physiology , *COMPARATIVE studies , *GAMMA rays , *GENES , *RESEARCH methodology , *MEDICAL cooperation , *ONCOGENES , *RESEARCH , *TIME , *EVALUATION research , *COLONY-forming units assay , *PHYSIOLOGICAL effects of radiation , *THERAPEUTICS - Abstract
Purpose: RH1 is a newly developed bioreductive agent, and its bioactivation is mediated by the enzyme DT-diaphorase (DTD). We have shown previously that RH1 is highly cytotoxic against cells expressing high DTD, using the p53-mutated MDA231 human breast cancer cell line transfected with the DTD gene (D7 cells). We now report that both RH1 and gamma-irradiation cause D7 cells to arrest in the G2/M cell cycle phase and undergo polyploidy. The latter is a way of p53-mutated cells responding to DNA-damaging agents. Only a small proportion of the polyploid cells are clonogenic, hence polyploidy may contribute to the reproductive failure of the cells after RH1 and irradiation. Thus, we investigated the effect of RH1 and gamma-irradiation on the formation of polyploid cells and a sub-G1 population (as a measure of apoptosis) in relation to the G2/M cell cycle block.Methods and Materials: MDA231 D7 cells were treated using a range of RH1 doses. The cells were irradiated using 2 Gy or 5 Gy gamma-rays either as a single dose or in combination with RH1. An IC(90) dose (dose to kill 90% of the cells) of RH1 was administered for 3 h followed by irradiation after a further 24 h. Subsequent changes in cell cycle and polyploidy (DNA content in excess of that of G2/M cells) were examined.Results: Treatment of D7 cells with the RH1 resulted in 60-70% of cells arrested in the G2/M phase of the cell cycle by 24 h, which decreased to control levels by 48 h. Irradiation with 2 Gy and 5 Gy caused a similar G2/M block at 12-24 h, which was followed by a sharp decline at 24-48 h. In contrast, the same dose of radiation combined with RH1 held the cells in the G2/M phase up to 48 h, and this pattern reached pretreatment levels at 72-96 h. Most control cells were found to contain a small number of spontaneously arising polyploid cells. The development of polyploid cells was evident from 12 h after all treatments and showed a significant increase at 48 h and subsequently. As opposed to this, apoptosis measured by the sub-G1 cell population in DNA analyses showed a tendency to increase according to the elapsed time for each group of treatments. Single treatments with RH1 caused a significant increase in the apoptotic population between 48 and 120 h. The first significant increase in apoptosis was observed at 48 h for 5 Gy, 2 Gy + RH1, and 5 Gy + RH1 treatments, and showed a tendency to increase further at later times, but the 2 Gy dose gave an earlier apoptotic peak at 24 h, which decreased to 96 h. The addition of RH1 to the irradiation did not increase the formation of polyploid cells or apoptosis compared with radiation alone (2 Gy vs. RH1 + 2 Gy or 5 Gy vs. RH1 + 5 Gy). The higher dose of irradiation (5 Gy vs. 2 Gy) resulted in a significantly higher proportion of polyploid cells (but not of apoptotic cells) when used alone or in combination (5 Gy + RH1 vs. 2 Gy + RH1).Conclusions: Both RH1 and gamma-irradiation, individually and in combination, showed a significant G2/M block in MDA231 D7 breast cancer cells. The formation of polyploid cells was dependent more on the radiation dose rather than on the pretreatment with RH1. The polyploid cell population was observed after the G2/M cell cycle phase arrest, and it preceded the late increase of the apoptotic cell population. The role of polyploidy in cell reproductive failure in the total cell population is not known, but it appears to contribute to cytotoxicity in cells released from the G2/M cell cycle phase block. [ABSTRACT FROM AUTHOR]- Published
- 2004
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23. 3-Substituted-5-aziridinyl-1-methylindole-4,7-diones as NQO1-directed antitumour agents: mechanism of activation and cytotoxicity in vitro
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Jaffar, Mohammed, Phillips, Roger M., Williams, Kaye J., Mrema, Ibrahim, Cole, Christian, Wind, Natasha S., Ward, Timothy H., Stratford, Ian J., and Patterson, Adam V.
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ANTINEOPLASTIC agents , *HYPOXEMIA , *ALDEHYDES , *MITOMYCIN C - Abstract
Indolequinone agents are a unique class of bioreductive cytotoxins that can function as dual substrates for both one- and two-electron reductases. This endows them with the potential to be either hypoxia-selective cytotoxins or NAD(P)H:quinone oxidoreductase 1 (NQO1)-directed prodrugs, respectively. We have studied the structure–activity relationships of four novel indolequinone analogues with regard to one- and/or two-electron activation. Single-electron metabolism was achieved by exposing the human carcinoma cell line T47D to each agent under hypoxic conditions, whilst concerted two-electron metabolism was assessed by stably expressing the cDNA for human NQO1 in a cloned cell line of T47D. The C-3 and C-5 positions of the indolequinone nucleus were modified to manipulate reactivity of the reduction products and the four prodrugs were identified as NQO1 substrates of varying specificity. Two of the four prodrugs, in which both C-3 and C-5 groups remained functional, proved to be NQO1-directed cytotoxins with selectivity ratios of 60- to 80-fold in the T47D (WT) versus the NQO1 overexpressing T47D cells. They also retained selectivity as hypoxic cytotoxins with oxic/hypoxic ratios of 20- to 22-fold. Replacement of the C-3 hydroxymethyl leaving group with an aldehyde group ablated all selectivity in air and hypoxia in both cell lines. Addition of a 2-methyl group on the C-5 aziridinyl group to introduce steric hinderance reduced but did not abolish NQO1-dependent metabolism. However, it enhanced single-electron metabolism-dependent DNA cross-linking in a manner that was independent of cytotoxicity. These data demonstrate that subtle structure–activity relationship exists for different cellular reductases and under certain circumstances distinct forms of DNA damage can arise, the cytotoxic consequences of which can vary. This study identifies a candidate indolequinone analogue for further development as a dual hypoxia and NQO1-directed prodrug. [Copyright &y& Elsevier]
- Published
- 2003
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24. Synthesis and antiproliferative activity of C- and N-terminal analogues of culicinin D.
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Li, Freda F., Stubbing, Louise A., Kavianinia, Iman, Abbattista, Maria R., Harris, Paul W.R., Smaill, Jeff B., Patterson, Adam V., and Brimble, Margaret A.
- Abstract
Culicinin D (1), a 10 amino acid peptaibol containing several unusual residues, has been shown to exhibit potent anticancer activity. Previous work in our group towards developing a structure-activity relationship (SAR) for this peptaibol has concentrated on replacement of the synthetically challenging AHMOD (3) and AMD (4) residues, resulting in the discovery of analogues with equivalent or better potency and simplified synthesis. The SAR of this peptaibol is extended in this work by investigating the effect of the N -terminal lipid tail and C -terminal amino alcohol, revealing the key contribution of each of these moieties on antiproliferative activity in a panel of breast and lung cancer cell lines. [ABSTRACT FROM AUTHOR]
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- 2020
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25. Alanine scan-guided synthesis and biological evaluation of analogues of culicinin D, a potent anticancer peptaibol.
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Kavianinia, Iman, Stubbing, Louise A., Abbattista, Maria R., Harris, Paul W.R., Smaill, Jeff B., Patterson, Adam V., and Brimble, Margaret A.
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BIOSYNTHESIS , *ALANINE , *AMINO acid residues , *AMINO acids , *STRUCTURE-activity relationships , *NATURAL products - Abstract
Culicinin D (1), a 10 amino acid peptaibol originally isolated from Culicinomyces clavisporus , exhibits potent activity against a range of cancer cell lines. Building on our previous work exploring the structure–activity relationship (SAR) of the unusual (2 S ,4 S ,6 R)-AHMOD residue, a series of analogues of culicinin D were prepared to further investigate the SAR of these peptaibols. Alanine scanning of a potent and readily accessible analogue 23 revealed the effect of each residue on antiproliferative activity, and a small panel of analogues were prepared to explore the SAR of the non-natural amino acid residue (2 S ,4 R)-AMD. Results from the alanine scan were used to design an expanded library of culicinin D analogues, leading to the discovery of cyclohexylalanine analogue 52 , which exhibited better antiproliferative activity than the natural product 1. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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26. 128: Prediction of Antitumor Activity of PR-104, a New Hypoxia Activated Mustard, Using Measurements of DNA Interstrand Crosslinks by the Comet Assay
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Dorie, Mary Jo, Ahn, G-one, Menke, Douglas, Douglas, Rachelle, Siim, Bronwyn, Patterson, Adam, Wilson, William, and Brown, Martin
- Published
- 2006
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