In vitro and in vivo studies using BW12C: toxicity, haemoglobin modification and effects on the radiosensitivity of normal marrow and RIF-1 tumours in mice.
BW12C binds to haemoglobin, shifting the oxygen saturation curve to the left, and is under investigation as an inducer of tumour hypoxia. The intrinsic cellular toxicity of the drug to RIF-1 and EMT6 cells in monolayer culture was studied, and IC50 values of 100 micrograms ml-1 for 24 h exposure and 10 micrograms ml-1 for 4-day exposure were measured. The LD50 (95% CL) in C3H mice was shown to be 124 (118-130) mg kg-1 for normal, rapid i.v. injection of the drug, and 173 (164-181) mg kg-1 for slow injection. The well-tolerated dose of 70 mg kg-1, used for all subsequent studies, was shown to produce a maximum haemoglobin modification of 70% 5 min after i.v. administration. This effect decayed with a half-life (+/- 2 se) of 76 +/- 8 min, giving 50% modification at 30 min and 22-25% modification at 2 h after administration. A dose of 70 mg kg-1 BW12C administered 30 min before irradiation protected animals against lethality, and increased the radiation LD50 (95% CL) from 7.16 (7.05-7.27) to 7.86 (7.70-8.02) Gy, representing a DMF of 1.1. In contrast the same drug dose and schedule did not alter normal marrow CFUs radiosensitivity at doses up to 6 Gy. The dose of 70 mg kg-1 did, however, cause marked radioprotection in RIF-1 intramuscular leg tumours. Four- to seven-fold increases in survival were measured by clonogenic cell survival immediately or 24 h after treatment. Protection was maximal 15 to 30 min after administration, and absent by 2 h. The drug did not protect RIF-1 cells in culture against radiation damage, indicating that the in vivo effect is indirect. BW12C is therefore an effective tumour radioprotector in this tumour model, in a manner consistent with an increase in tumour hypoxic fraction, although factors other than changes in blood chemistry may also be involved.
MRC Unit, MRC Centre, Cambridge, UK.
D J
DJ
Honess
A B
AB
Nethersell
N M
NM
Bleehen
D000818:Animals; D001547:Benzaldehydes / Q000627:therapeutic use* / Q000633:toxicity; D002470:Cell Survival / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D005260:Female; D006454:Hemoglobins / Q000187:drug effects*; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000188:drug therapy*; D010100:Oxygen / Q000097:blood; D011836:Radiation Tolerance / Q000187:drug effects*; D011837:Radiation-Protective Agents / Q000627:therapeutic use* / Q000633:toxicity
D001547:Benzaldehydes; D006454:Hemoglobins; D011837:Radiation-Protective Agents; C039599:5-(2-formyl-3-hydroxyphenoxy)pentanoic acid; D010100:Oxygen