Radiation Research

Published by: Radiation Research Society

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Radiation Research 168(2):218-225. 2007
doi: 10.1667/RR0962.1

The Effects of Efaproxyn™ (Efaproxiral) on Subcutaneous RIF-1 Tumor Oxygenation and Enhancement of Radiotherapy-Mediated Inhibition of Tumor Growth in Mice

Huagang Houa, Nadeem Khana, Oleg Y. Grinberga, Hongsheng Yua, Stalina A. Grinberga, Shiyi Lua, Eugene Demidenkob, Robert P. Steffenc, and Harold M. Swartz1a

aDepartment of Diagnostic Radiology, EPR Center for the Study of Viable Systems, Dartmouth Medical School, Hanover, New Hampshire 03755;

bDepartment of Community and Family Medicine, Section of Biostatistics and Epidemiology, Dartmouth Medical School, Lebanon, New Hampshire 03766;

cAllos Therapeutics, Inc., Westminster, Colorado 80020

1Address for correspondence: Department of Diagnostic Radiology, EPR Center for the Study of Viable Systems, Dartmouth Medical School, Hanover, NH 03755;

Abstract

Hou, H., Khan, N., Grinberg, O. Y., Yu, H., Grinberg, S. A., Lu, S., Demidenko, E., Steffen, R. P. and Swartz, H. M. The Effects of Efaproxyn™ (Efaproxiral) on Subcutaneous RIF-1 Tumor Oxygenation and Enhancement of Radiotherapy-Mediated Inhibition of Tumor Growth in Mice. Radiat. Res. 168, 218–225 (2007).

Efaproxiral, an allosteric modifier of hemoglobin, reduces hemoglobin-oxygen binding affinity, facilitating oxygen release from hemoglobin, which is likely to increase tissue pO2. The purpose of this study was to determine the effect of efaproxiral on tumor oxygenation and growth inhibition of RIF-1 tumors that received X radiation (4 Gy) plus oxygen breathing compared to radiation plus oxygen plus efaproxiral daily for 5 days. Two lithium phthalocyanine (LiPc) deposits were implanted in RIF-1 tumors in C3H mice for tumor pO2 measurements using EPR oximetry. Efaproxiral significantly increased tumor oxygenation by 8.4 to 43.4 mmHg within 5 days, with maximum increases at 22–31 min after treatment. Oxygen breathing alone did not affect tumor pO2. Radiation plus oxygen plus efaproxiral produced tumor growth inhibition throughout the treatment duration, and inhibition was significantly different from radiation plus oxygen from day 3 to day 5. The results of this study provide unambiguous quantitative information on the effectiveness of efaproxiral to consistently and reproducibly increase tumor oxygenation over the course of 5 days of treatment, modeling the clinical use of efaproxiral. Also, based on the tumor growth inhibition, the study shows the efaproxiral-enhanced tumor oxygenation was radiobiologically significant. This is the first study to demonstrate the ability of efaproxiral to increase tumor oxygenation and to increase the tumor growth inhibition of radiotherapy over 5 days of treatment.

Received: July 3, 2006; Accepted: January 16, 2007



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FIG. 1. Time course of tumor pO2 (mean ± SE, mmHg) measured using EPR in mice before and after treatment with efaproxiral (150 mg/kg, i.p.) and radiation (4 Gy/day). In each panel, the bold line shows the average effect for all animals in the group using an exponential quadratic function. Each thin line is the standard error from all implants (two implants/each mouse). Black arrows indicate the starting time for administering efaproxiral, while the dashed arrows indicate the starting time of irradiation. Efa + IR = efaproxiral + radiation; Veh + IR = vehicle + radiation

FIG. 2. Each line shows the average response for all animals in the treatment (efaproxiral + radiation) group using an exponential quadratic function. Tmax = time from end of irradiation to reach maximum pO2. ‡‡P < 0.01, compared with the maximum tumor pO2 on other days

FIG. 3. Tumor growth inhibition measurements in mice treated with efaproxiral. The tumors were injected with either 150 mg/kg efaproxiral or an equivalent volume of vehicle, i.p., and tumors were irradiated around 35 min later with a dose of 4 Gy. Day 0 is the day before efaproxiral or vehicle and radiation treatments. Data points are the means ± SE. ‡P < 0.01, *P <0.05, compared with efaproxiral/radiation; †P < 0.01, compared with vehicle/radiation (two-tailed unpaired t test). For reference we also show the results in unirradiated tumors (efaproxiral only) from a previous study (18) that noted the results but did not provide the detailed data

table

TABLE 1 Tumor pO2 (mmHg, mean ± SE) from Two Implants per Tumors after Daily Treatments of RIF-1 Tumors with Efaproxiral (150 mg/kg, i.p.) or Vehicle (same volume) Prior to X Irradiation (4 Gy) in Mice on 5 Consecutive Days

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