Low-density lipoprotein as a carrier of antitumoral drugs: in vivo fate of drug-human low-density lipoprotein complexes in mice.

Previous studies have demonstrated that human leukemic cells and certain cancer cells in culture have a higher uptake of plasma low-density lipoprotein (LDL) than the corresponding normal cells. Therefore LDL has been proposed as a drug carrier for anticancer agents. In the present investigation, we have developed a method to incorporate a lipophilic derivative of doxorubicin, N-trifluoroacetyladriamycin-14-valerate, into LDL. The method involves lyophilization of LDL in the presence of sucrose as protective agent and gives an N-trifluoroacetyladriamycin-14-valerate-LDL complex containing about 100 drug molecules per LDL particle. The in vivo fate of the complex in mice as judged from the disappearance from plasma and accumulation in organs was similar to that of native LDL. When cultured human fibroblasts were incubated with N-trifluoroacetyladriamycin-14-valerate-LDL, cellular drug accumulation was dependent on the LDL receptor activity of the cells. The covalent linkage of two anthracycline derivatives to lysine residues of LDL yielded conjugates with drug/LDL molar ratios ranging up to 80. With increasing substitution, there was a progressive decline in the affinity of the conjugate for the LDL receptor in vitro. The in vivo fate of such conjugates was quite similar to that of native LDL. We conclude that it is possible to associate cytotoxic agents with LDL without interfering with its in vivo behavior.

[1]  G. W. Anderson,et al.  Synthesis of N-hydroxysuccinimide esters of acyl peptides by the mixed anhydride method. , 1967, Journal of the American Chemical Society.

[2]  R. Mahley,et al.  Role of lysine residues of plasma lipoproteins in high affinity binding to cell surface receptors on human fibroblasts. , 1978, The Journal of biological chemistry.

[3]  C. McArdle,et al.  Low-density lipoprotein metabolism in mice with soft tissue tumours. , 1984, Biochimica et biophysica acta.

[4]  R. Mahley,et al.  Altered metabolism (in vivo and in vitro) of plasma lipoproteins after selective chemical modification of lysine residues of the apoproteins. , 1979, The Journal of clinical investigation.

[5]  M. Brown,et al.  Binding and degradation of low density lipoproteins by cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. , 1974, The Journal of biological chemistry.

[6]  M. Krieger,et al.  Reconstituted low density lipoprotein: a vehicle for the delivery of hydrophobic fluorescent probes to cells. , 1979, Journal of supramolecular structure.

[7]  M. Moore,et al.  A transplantable myelomonocytic leukemia in BALB-c mice: cytology, karyotype, and muramidase content. , 1969, Journal of the National Cancer Institute.

[8]  F. Zunino,et al.  Biologic Activity of Daunorubicin Linked to Proteins via the Methylketone Side Chain , 1981, Tumori.

[9]  E. Simpson,et al.  Low-density lipoprotein as a potential vehicle for chemotherapeutic agents and radionucleotides in the management of gynecologic neoplasms. , 1981, American journal of obstetrics and gynecology.

[10]  W. Mantulin,et al.  Incorporation of defined cholesteryl esters into lipoproteins using cholesteryl ester-rich microemulsions. , 1982, The Journal of biological chemistry.

[11]  G. Gahrton,et al.  Significance of the low-density lipoprotein (LDL) receptor pathway for the in vitro accumulation of AD-32 incorporated into LDL in normal and leukemic white blood cells. , 1984, Cancer treatment reports.

[12]  M. Masquelier,et al.  A covalent linkage between daunorubicin and proteins that is stable in serum and reversible by lysosomal hydrolases, as required for a lysosomotropic drug-carrier conjugate: in vitro and in vivo studies. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[13]  F. Zunino,et al.  Interaction of some daunomycin derivatives with deoxyribonucleic acid and their biological activity. , 1971, Biochemical pharmacology.

[14]  S. Yanovich,et al.  Preparation and interaction of a low-density lipoprotein:daunomycin complex with P388 leukemic cells. , 1984, Cancer research.

[15]  R. Levy,et al.  The metabolism of low density lipoprotein in familial type II hyperlipoproteinemia. , 1972, The Journal of clinical investigation.

[16]  G. Canti,et al.  In vivo assimilation of low density lipoproteins by a fibrosarcoma tumour line in mice. , 1984, Cancer letters.

[17]  R. Juliano,et al.  Liposomes as a drug delivery system , 1981 .

[18]  B. Lundberg,et al.  Preparation of biologically active analogs of serum low density lipoprotein. , 1984, Journal of lipid research.

[19]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[20]  G. Gahrton,et al.  HYPOCHOLESTEROLAEMIA IN MALIGNANCY DUE TO ELEVATED LOW-DENSITY-LIPOPROTEIN-RECEPTOR ACTIVITY IN TUMOUR CELLS: EVIDENCE FROM STUDIES IN PATIENTS WITH LEUKAEMIA , 1985, The Lancet.

[21]  J. Lovelock,et al.  The denaturation of lipid-protein complexes as a cause of damage by freezing , 1957, Proceedings of the Royal Society of London. Series B - Biological Sciences.

[22]  M. Brown,et al.  Low-density lipoprotein (LDL) receptor activity in human acute myelogenous leukemia cells. , 1978, Blood.

[23]  M. Masquelier,et al.  Amino acid and dipeptide derivatives of daunorubicin. 1. Synthesis, physicochemical properties, and lysosomal digestion. , 1980, Journal of medicinal chemistry.

[24]  V. P. Collins,et al.  Delivery of aclacinomycin A to human glioma cells in vitro by the low-density lipoprotein pathway. , 1983, Cancer research.

[25]  M. Brown,et al.  Receptor-mediated endocytosis: insights from the lipoprotein receptor system. , 1979, Proceedings of the National Academy of Sciences of the United States of America.