Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia

Paul Smith, Marie Wiltshire, Heather Nesbitt, Niall Byrne, Louise Ming, Declan McKenna, Jenny Worthington, Stephanie McKeown, Laurence Patterson, Rachel Errington

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Background: Tumour microenvironment hypoxia (TMH)facilitates disease progression and therapeutic resistance,presenting challenges for the treatment of prostate and pancreaticcancer. The non-toxic anthraquinone prodrug OCT1002(OncoTherics, UK) is designed to distribute widely in tissues,become irreversibly activated to OCT1001, a potent DNAtopoisomerase II inhibitor, thereby acting as unidirectionalhypoxia activated prodrug (uHAP). We have investigated therelationship between intracellular generation and persistence ofTMH-generated OCT1001 and biological responses both in vitroand in vivo.Methods: Confocal imaging and FCM exploited the intrinsic farredfluorescence of the uHAP. Cancer cell lines were exposed toclinically relevant prodrug levels at 1-100 nM under hypoxia (4d;1-3% oxygen); studied for cell proliferation, OCT1002bioreduction and cell cycle arrest. Human prostate cancerxenografts expressing a luciferase reporter (LNCaP-luc) in Balb/cSCID mice were used to monitor growth delay, lung metastasisand vascular changes in relation to OCT1002 targeting in dorsalskin-flap/window chambers. pO2 of tumours was recorded usingan Oxylite 2000 system and assessed in tissues by Glut1 staining.Xenograft induced hypoxia was achieved by exposure tobicalutamide (Casodex™; a non-steroidal anti-androgen).Pancreatic cancer human BX-PC3/SCID mouse xenografts,presenting intrinsic hypoxia, were used to assess uHAPmonotherapy.Results: Hypoxia-dependent growth arrest induced by OCT1002was found in a wide range cancer cell lines irrespective of p53status. Cytostasis and sustained G2 cell cycle arrest was achievedwithin 1 population doubling at 100 nM [% cells arrested at1%>3%>>air]. OCT1002 and hypoxia co-exposure generatedpersistent intracellular fluorescence attributable to the metaboliteOCT1001 while the prodrug OCT1002 was not retained.OCT1001 located within LNCaP-luc tumours at regions distantfrom blood vessels, persisted for >7 days, slowed tumour growthand reduced lung metastasis. A pilot study showed that a singledose OCT1002 induced a growth delay in pancreatic cancer BXPC3/SCIDmice xenografts, which are hypoxic, comparable withthat achieved by fractionated Gemcitabine treatment.Conclusion: Cytometric analyses have shown effective OCT1002activation in vitro. A quantitative relationship was found betweenthe degree of hypoxia, extent of uHAP bioreduction, persistenceof OCT1001, growth delay and first cycle G2 arrest. In vivoimaging, in a prostate cancer model that recapitulates TMHinduced by anti-androgen treatment, showed that uHAPactivation at TMH locations. The slowing of tumour growth andreduction in metastatic potential in prostate cancer together withmonotherapy efficacy in a pancreatic cancer model suggest thatOCT1002 has potential for tumour indications and in combinedmodalities where TMH is considered to be a bar
LanguageEnglish
Title of host publicationUnknown Host Publication
Pages128
Number of pages268
Publication statusPublished - Jun 2015
EventCYTO 2015, the 30th Congress of the International Society for Advancement of Cytometry - Glasgow,Scotland
Duration: 1 Jun 2015 → …

Conference

ConferenceCYTO 2015, the 30th Congress of the International Society for Advancement of Cytometry
Period1/06/15 → …

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Prodrugs
Tumor Microenvironment
Pancreatic Neoplasms
Neoplasms
Heterografts
Growth
gemcitabine
Androgens
Blood Vessels
Prostate
Prostatic Neoplasms
G2 Phase Cell Cycle Checkpoints
Cell Line
Anthraquinones
Lung
SCID Mice
Cell Cycle Checkpoints
Tumor Hypoxia
In Vitro Techniques
Luciferases

Keywords

  • OCT1002
  • uHAP
  • Prostate Cancer
  • Hypoxia

Cite this

Smith, P., Wiltshire, M., Nesbitt, H., Byrne, N., Ming, L., McKenna, D., ... Errington, R. (2015). Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia. In Unknown Host Publication (pp. 128)
Smith, Paul ; Wiltshire, Marie ; Nesbitt, Heather ; Byrne, Niall ; Ming, Louise ; McKenna, Declan ; Worthington, Jenny ; McKeown, Stephanie ; Patterson, Laurence ; Errington, Rachel. / Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia. Unknown Host Publication. 2015. pp. 128
@inproceedings{76ac94d111d842819030b7b6045e3654,
title = "Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia",
abstract = "Background: Tumour microenvironment hypoxia (TMH)facilitates disease progression and therapeutic resistance,presenting challenges for the treatment of prostate and pancreaticcancer. The non-toxic anthraquinone prodrug OCT1002(OncoTherics, UK) is designed to distribute widely in tissues,become irreversibly activated to OCT1001, a potent DNAtopoisomerase II inhibitor, thereby acting as unidirectionalhypoxia activated prodrug (uHAP). We have investigated therelationship between intracellular generation and persistence ofTMH-generated OCT1001 and biological responses both in vitroand in vivo.Methods: Confocal imaging and FCM exploited the intrinsic farredfluorescence of the uHAP. Cancer cell lines were exposed toclinically relevant prodrug levels at 1-100 nM under hypoxia (4d;1-3{\%} oxygen); studied for cell proliferation, OCT1002bioreduction and cell cycle arrest. Human prostate cancerxenografts expressing a luciferase reporter (LNCaP-luc) in Balb/cSCID mice were used to monitor growth delay, lung metastasisand vascular changes in relation to OCT1002 targeting in dorsalskin-flap/window chambers. pO2 of tumours was recorded usingan Oxylite 2000 system and assessed in tissues by Glut1 staining.Xenograft induced hypoxia was achieved by exposure tobicalutamide (Casodex™; a non-steroidal anti-androgen).Pancreatic cancer human BX-PC3/SCID mouse xenografts,presenting intrinsic hypoxia, were used to assess uHAPmonotherapy.Results: Hypoxia-dependent growth arrest induced by OCT1002was found in a wide range cancer cell lines irrespective of p53status. Cytostasis and sustained G2 cell cycle arrest was achievedwithin 1 population doubling at 100 nM [{\%} cells arrested at1{\%}>3{\%}>>air]. OCT1002 and hypoxia co-exposure generatedpersistent intracellular fluorescence attributable to the metaboliteOCT1001 while the prodrug OCT1002 was not retained.OCT1001 located within LNCaP-luc tumours at regions distantfrom blood vessels, persisted for >7 days, slowed tumour growthand reduced lung metastasis. A pilot study showed that a singledose OCT1002 induced a growth delay in pancreatic cancer BXPC3/SCIDmice xenografts, which are hypoxic, comparable withthat achieved by fractionated Gemcitabine treatment.Conclusion: Cytometric analyses have shown effective OCT1002activation in vitro. A quantitative relationship was found betweenthe degree of hypoxia, extent of uHAP bioreduction, persistenceof OCT1001, growth delay and first cycle G2 arrest. In vivoimaging, in a prostate cancer model that recapitulates TMHinduced by anti-androgen treatment, showed that uHAPactivation at TMH locations. The slowing of tumour growth andreduction in metastatic potential in prostate cancer together withmonotherapy efficacy in a pancreatic cancer model suggest thatOCT1002 has potential for tumour indications and in combinedmodalities where TMH is considered to be a bar",
keywords = "OCT1002, uHAP, Prostate Cancer, Hypoxia",
author = "Paul Smith and Marie Wiltshire and Heather Nesbitt and Niall Byrne and Louise Ming and Declan McKenna and Jenny Worthington and Stephanie McKeown and Laurence Patterson and Rachel Errington",
year = "2015",
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language = "English",
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Smith, P, Wiltshire, M, Nesbitt, H, Byrne, N, Ming, L, McKenna, D, Worthington, J, McKeown, S, Patterson, L & Errington, R 2015, Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia. in Unknown Host Publication. pp. 128, CYTO 2015, the 30th Congress of the International Society for Advancement of Cytometry, 1/06/15.

Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia. / Smith, Paul; Wiltshire, Marie; Nesbitt, Heather; Byrne, Niall; Ming, Louise; McKenna, Declan; Worthington, Jenny; McKeown, Stephanie; Patterson, Laurence; Errington, Rachel.

Unknown Host Publication. 2015. p. 128.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - Cytometry of Anticancer Prodrug OCT1002 Activation and Targeting Using In Vitro and In Vivo Models of Tumour Hypoxia

AU - Smith, Paul

AU - Wiltshire, Marie

AU - Nesbitt, Heather

AU - Byrne, Niall

AU - Ming, Louise

AU - McKenna, Declan

AU - Worthington, Jenny

AU - McKeown, Stephanie

AU - Patterson, Laurence

AU - Errington, Rachel

PY - 2015/6

Y1 - 2015/6

N2 - Background: Tumour microenvironment hypoxia (TMH)facilitates disease progression and therapeutic resistance,presenting challenges for the treatment of prostate and pancreaticcancer. The non-toxic anthraquinone prodrug OCT1002(OncoTherics, UK) is designed to distribute widely in tissues,become irreversibly activated to OCT1001, a potent DNAtopoisomerase II inhibitor, thereby acting as unidirectionalhypoxia activated prodrug (uHAP). We have investigated therelationship between intracellular generation and persistence ofTMH-generated OCT1001 and biological responses both in vitroand in vivo.Methods: Confocal imaging and FCM exploited the intrinsic farredfluorescence of the uHAP. Cancer cell lines were exposed toclinically relevant prodrug levels at 1-100 nM under hypoxia (4d;1-3% oxygen); studied for cell proliferation, OCT1002bioreduction and cell cycle arrest. Human prostate cancerxenografts expressing a luciferase reporter (LNCaP-luc) in Balb/cSCID mice were used to monitor growth delay, lung metastasisand vascular changes in relation to OCT1002 targeting in dorsalskin-flap/window chambers. pO2 of tumours was recorded usingan Oxylite 2000 system and assessed in tissues by Glut1 staining.Xenograft induced hypoxia was achieved by exposure tobicalutamide (Casodex™; a non-steroidal anti-androgen).Pancreatic cancer human BX-PC3/SCID mouse xenografts,presenting intrinsic hypoxia, were used to assess uHAPmonotherapy.Results: Hypoxia-dependent growth arrest induced by OCT1002was found in a wide range cancer cell lines irrespective of p53status. Cytostasis and sustained G2 cell cycle arrest was achievedwithin 1 population doubling at 100 nM [% cells arrested at1%>3%>>air]. OCT1002 and hypoxia co-exposure generatedpersistent intracellular fluorescence attributable to the metaboliteOCT1001 while the prodrug OCT1002 was not retained.OCT1001 located within LNCaP-luc tumours at regions distantfrom blood vessels, persisted for >7 days, slowed tumour growthand reduced lung metastasis. A pilot study showed that a singledose OCT1002 induced a growth delay in pancreatic cancer BXPC3/SCIDmice xenografts, which are hypoxic, comparable withthat achieved by fractionated Gemcitabine treatment.Conclusion: Cytometric analyses have shown effective OCT1002activation in vitro. A quantitative relationship was found betweenthe degree of hypoxia, extent of uHAP bioreduction, persistenceof OCT1001, growth delay and first cycle G2 arrest. In vivoimaging, in a prostate cancer model that recapitulates TMHinduced by anti-androgen treatment, showed that uHAPactivation at TMH locations. The slowing of tumour growth andreduction in metastatic potential in prostate cancer together withmonotherapy efficacy in a pancreatic cancer model suggest thatOCT1002 has potential for tumour indications and in combinedmodalities where TMH is considered to be a bar

AB - Background: Tumour microenvironment hypoxia (TMH)facilitates disease progression and therapeutic resistance,presenting challenges for the treatment of prostate and pancreaticcancer. The non-toxic anthraquinone prodrug OCT1002(OncoTherics, UK) is designed to distribute widely in tissues,become irreversibly activated to OCT1001, a potent DNAtopoisomerase II inhibitor, thereby acting as unidirectionalhypoxia activated prodrug (uHAP). We have investigated therelationship between intracellular generation and persistence ofTMH-generated OCT1001 and biological responses both in vitroand in vivo.Methods: Confocal imaging and FCM exploited the intrinsic farredfluorescence of the uHAP. Cancer cell lines were exposed toclinically relevant prodrug levels at 1-100 nM under hypoxia (4d;1-3% oxygen); studied for cell proliferation, OCT1002bioreduction and cell cycle arrest. Human prostate cancerxenografts expressing a luciferase reporter (LNCaP-luc) in Balb/cSCID mice were used to monitor growth delay, lung metastasisand vascular changes in relation to OCT1002 targeting in dorsalskin-flap/window chambers. pO2 of tumours was recorded usingan Oxylite 2000 system and assessed in tissues by Glut1 staining.Xenograft induced hypoxia was achieved by exposure tobicalutamide (Casodex™; a non-steroidal anti-androgen).Pancreatic cancer human BX-PC3/SCID mouse xenografts,presenting intrinsic hypoxia, were used to assess uHAPmonotherapy.Results: Hypoxia-dependent growth arrest induced by OCT1002was found in a wide range cancer cell lines irrespective of p53status. Cytostasis and sustained G2 cell cycle arrest was achievedwithin 1 population doubling at 100 nM [% cells arrested at1%>3%>>air]. OCT1002 and hypoxia co-exposure generatedpersistent intracellular fluorescence attributable to the metaboliteOCT1001 while the prodrug OCT1002 was not retained.OCT1001 located within LNCaP-luc tumours at regions distantfrom blood vessels, persisted for >7 days, slowed tumour growthand reduced lung metastasis. A pilot study showed that a singledose OCT1002 induced a growth delay in pancreatic cancer BXPC3/SCIDmice xenografts, which are hypoxic, comparable withthat achieved by fractionated Gemcitabine treatment.Conclusion: Cytometric analyses have shown effective OCT1002activation in vitro. A quantitative relationship was found betweenthe degree of hypoxia, extent of uHAP bioreduction, persistenceof OCT1001, growth delay and first cycle G2 arrest. In vivoimaging, in a prostate cancer model that recapitulates TMHinduced by anti-androgen treatment, showed that uHAPactivation at TMH locations. The slowing of tumour growth andreduction in metastatic potential in prostate cancer together withmonotherapy efficacy in a pancreatic cancer model suggest thatOCT1002 has potential for tumour indications and in combinedmodalities where TMH is considered to be a bar

KW - OCT1002

KW - uHAP

KW - Prostate Cancer

KW - Hypoxia

M3 - Conference contribution

SP - 128

BT - Unknown Host Publication

ER -