Covalent coupling of methotrexate to dextran enhances the penetration of cytotoxicity into a tissue-like matrix.
For antitumor agents introduced directly into the intracranial space, the extent of penetration into tissue, and hence the effectiveness of therapy, depends on the rate of drug elimination from the tissue. To test the hypothesis that slowly eliminated agents would penetrate further through tissues, methotrexate (MTX)-dextran conjugates were produced by covalently linking MTX to dextran through a short-lived ester bond (MTX-ester-dextran; t1/2 approximately 3 days in buffered saline) and a longer-lived amide bond (MTX-amide-dextran; t1/2 > 20 days in buffered saline). The ability of these agents to kill cells and to penetrate through tissue was evaluated using: (a) human brain tumor (H80) cells in a standard format; (b) H80 cells in a novel three-dimensional format that mimics many characteristics of intracranial tumors; and (c) 9L gliosarcoma in the rat brain. Penetration into three-dimensional tissue-like matrices was performed by suspending H80 cells in agarose gels within a hollow fiber that was permeable to MTX but not dextran and injecting MTX or MTX-dextran conjugates into one end of the fiber. The cytotoxicity of MTX-ester-dextran and MTX-amide-dextran against H80 was equivalent to unmodified MTX (50% inhibitory concentration, approximately 0.01 microgram/ml). When released from a biodegradable polyanhydride polymer matrix, MTX and MTX-dextran conjugates retained their ability to inhibit dihydrofolate reductase activity. When MTX or MTX-dextran was diffused into the three-dimensional tumor cell matrix for 10 days, cytotoxic activity penetrated > 2 cm for MTX-amide-dextran and approximately 1 cm for MTX or MTX-ester-dextran; this enhanced penetration correlated with the stability of the MTX-dextran linkage. Intracranial polymeric delivery of MTX or MTX-amide-dextran to rats with intracranial 9L gliosarcoma produced modest but significant increases in survival; conjugation of MTX to dextran appeared to shift the dose-response curve to a lower dosage.
Dang
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Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Colvin
O M
OM
Brem
H
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Saltzman
W M
WM
D000818:Animals; D001932:Brain Neoplasms / Q000188:drug therapy*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D003692:Delayed-Action Preparations; D003911:Dextrans / Q000378:metabolism* / Q000627:therapeutic use / Q000633:toxicity*; D004058:Diffusion; D005910:Glioma; D018316:Gliosarcoma / Q000188:drug therapy*; D006801:Humans; D008727:Methotrexate / Q000031:analogs & derivatives / Q000378:metabolism / Q000627:therapeutic use / Q000633:toxicity*; D051381:Rats; D014407:Tumor Cells, Cultured
D003692:Delayed-Action Preparations; D003911:Dextrans; D008727:Methotrexate