Stability of cytotoxic luteinizing hormone-releasing hormone conjugate (AN-152) containing doxorubicin 14-O-hemiglutarate in mouse and human serum in vitro: implications for the design of preclinical studies.
Recently, we developed a series of cytotoxic peptide conjugates containing 14-O-glutaryl esters of doxorubicin (DOX) or 2-pyrrolino-DOX (AN-201). Serum carboxylesterase enzymes (CE) can partially hydrolyze these conjugates in the circulation, releasing the cytotoxic radical, before the targeting is complete. CE activity in serum of nude mice is about 10 times higher than in human serum. Thus, we found that the t(1/2) of AN-152, an analog of luteinizing hormone-releasing hormone (LH-RH) containing DOX, at 0.3 mg/ml is 19. 49 +/- 0.74 min in mouse serum and 126.06 +/- 3.03 min in human serum in vitro. The addition of a CE inhibitor, diisopropyl fluorophosphate (DFP), to mouse serum in vitro significantly (P < 0. 01) prolongs the t(1/2) of AN-152 to 69.63 +/- 4.44 min. When DFP is used in vivo, 400 nmol/kg cytotoxic somatostatin analog AN-238 containing AN-201 is well tolerated by mice, whereas all animals die after the same dose without DFP. In contrast, DFP has no effect on the tolerance of AN-201. A better tolerance to AN-238 after DFP treatment is due to the selective uptake of AN-238 by somatostatin receptor-positive tissues. Our results demonstrate that the suppression of the CE activity in nude mice greatly decreases the toxicity of cytotoxic hybrids containing 2-pyrrolino-DOX 14-O-hemiglutarate and brings this animal model closer to the conditions that exist in humans. The use of DFP together with these peptide conjugates in nude mice permits a better understanding of their mechanism of action and improves the clinical predictability of the oncological and toxicological results.
Nagy
A
A
Endocrine, Polypeptide, and Cancer Institute, Veterans Affairs Medical Center, New Orleans, LA 70112-2699, USA.
Plonowski
A
A
Schally
A V
AV
D000328:Adult; D000818:Animals; D000970:Antineoplastic Agents / Q000097:blood* / Q000737:chemistry; D001839:Bombesin / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000097:blood; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D004353:Drug Evaluation, Preclinical; D004347:Drug Interactions; D004355:Drug Stability; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D006207:Half-Life; D006801:Humans; D006868:Hydrolysis; D007531:Isoflurophate / Q000494:pharmacology; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009596:Nitrophenols / Q000494:pharmacology; D010261:Paraoxon / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology / Q000633:toxicity; D015431:Weight Loss / Q000187:drug effects
C110845:AN 238; D000970:Antineoplastic Agents; D002800:Cholinesterase Inhibitors; D009596:Nitrophenols; D011758:Pyrroles; D007531:Isoflurophate; C098751:AN 204; C104648:AN 207; C080883:LHRH, lysine(6)-doxorubicin; D007987:Gonadotropin-Releasing Hormone; C002887:bis(4-nitrophenyl)phosphate; D004317:Doxorubicin; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D001839:Bombesin; D010261:Paraoxon