The pathogenesis of vacuoles produced in rat and mouse liver cells by a conjugate of adenine arabinoside monophosphate with lactosaminated albumin

doi:10.1016/0168-8278(92)90062-T

Journal of Hepatology

Volume 15, Issue 3, July 1992, Pages 314-322

https://doi.org/10.1016/0168-8278(92)90062-T Get rights and content

Abstract

A conjugate of adenine arabinoside monophosphate with lactosaminated albumin produced vacuoles in hepatic cells of rats and mice when given at doses 5–10 times higher than that (35 mg/kg) capable of inhibiting hepatitis B virus replication in patients with chronic hepatitis B. The vacuoles were due to the swelling of secondary lysosomes probably caused by incapacity of the lysosomal enzymes to rapidly digest large amounts of conjugate into products able to cross the lysosomal membrane. Although vacuoles progressively disappeared when conjugate administration was discontinued, the present observation suggests caution in giving the conjugate to man at daily doses higher than 35 mg/kg.

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Cited by (19)

Lactosaminated human albumin, a hepatotropic carrier of drugs
2010, European Journal of Pharmaceutical Sciences
A selective delivery of drugs to liver can be obtained by conjugation with galactosyl terminating macromolecules. The conjugates selectively enter hepatocytes after interaction of the carrier galactose residues with the asialoglycoprotein receptor (ASGP-R) present only on these cells. Within hepatocytes the conjugates are transported to lysosomes where the drug is set free from the carrier, becoming concentrated in liver cells. The present article reviews the liver targeting of drugs obtained with lactosaminated albumin (L-SA), a neoglycoprotein exposing galactosyl residues. We report: (1) experiments which demonstrate the antiviral efficacy of the L-H(human)SA-ara-AMP conjugate in laboratory animals and in humans with viral hepatitis; (2) the property of a L-HSA conjugate with fluorodeoxyuridine to produce concentrations of the drug higher in hepatic sinusoids than in systemic circulation, with the potential of accomplishing a loco-regional, noninvasive treatment of liver micrometastases; (3) the increased anticancer activity of doxorubicin (DOXO) when coupled to L-HSA on all the forms of chemically induced rat hepatocellular carcinomas including those which do not express the ASGP-R.
A new peptide ligand that targets particles and heterologous proteins to hepatocytes in vivo [1]
2003, Molecular Therapy
Coupling of 5-fluoro 2′-deoxyuridine to lactosaminated poly-L-lysine: An approach to a regional, non-invasive chemotherapy of liver micrometastases
2001, Biochemical Pharmacology
Citation Excerpt :
One hour after i.v. administration of l-poly[3H](LYS)–FUdR (14.7 μg/g, corresponding to 5 μg FUdR and 39,000 dpm/g), radioactivity values per mg cell proteins measured in parenchymal and sinusoidal cells isolated by collagenase perfusion (see Materials and methods) were 2052 ± 167 and 1641 ± 116, respectively. Penetration of a galactosyl-terminating conjugate into liver sinusoidal cells was previously observed in rats injected with l-HSA–ara-A [20,23]. The mechanism of entry of l-poly(LYS)–FUdR and l-HSA–ara-A into sinusoidal cells is not known; it might be a consequence of a non-specific adsorption endocytosis.
Nucleoside analogs conjugated with galactosyl-terminating peptides selectively enter liver cells and after intracellular release from the carrier partly exit into bloodstream, resulting in higher concentrations in liver blood than in systemic circulation. The aim of the present experiments was to ascertain whether, in mice injected with non-toxic doses of a 5-fluoro 2′-deoxyuridine (FUdR) conjugate with lactosaminated poly-l-lysine (l-poly(LYS)), the drug was released by hepatic cells in high enough amounts to be pharmacologically active on neoplastic cells infiltrating the liver. We observed that l-poly(LYS)–FUdR inhibited the growth of hepatic metastases induced by intrasplenic administration of murine colon carcinoma C-26 cells. l-poly(LYS)–FUdR was not toxic for C-26 cells in vitro, was selectively taken up by mouse liver, and was stable in mouse blood, indicating that the effect on the metastases was due to FUdR (and/or its active metabolites) released in liver blood after the conjugate was taken up by the hepatic cells. These results suggest that l-poly(LYS)–FUdR might be useful in adjuvant chemotherapy of tumors giving liver metastases. The drug released from hepatic cells into liver blood following conjugate administration via the peripheral venous route might accomplish a locoregional, non-invasive treatment of micrometastases nourished by liver sinusoids.
Hepatotropic conjugate of adenine arabinoside monophosphate with lactosaminated poly-L-lysine. Synthesis of the carrier and pharmacological properties of the conjugate
1997, Journal of Hepatology
Background/Aims: The hepatotropic conjugate of adenine arabinoside monophosphate with lactosaminated poly-l-lysine (l-Poly(Lys)) must have a high solubility in order to be injected in a small volume compatible with the intramuscular route. In this paper the molecular weights of Poly(Lys) which allowed the synthesis of conjugates with the properties of high solubility and limited loss by the kidney were determined and a procedure for obtaining Poly(Lys) preparations with the required range of polymerization has been described.
Methods: Conjugates were prepared using Poly(Lys) of different molecular weights obtained by the procedure described here or purchased from a commercial source. Their solubility and renal loss in mice was determined.
Results: Poly(Lys) with molecular weights ranging from 45000 and 65000 Da guarantees high solubility and low renal elimination of the conjugates. Conjugate preparations with these properties, intramuscularly administered to woodchuch hepatitis virus-infected woodchucks for 37 days at a daily dose of 5.8 mg/kg exerted a strong antiviral activity. These preparations were devoid of acute toxicity in rat and caused no toxic effects when injected intramuscularly daily for 28 days at a dose ten times higher than that active in woodchucks.
Conclusions: The results support the possibility of a clinical use of l-Poly(Lys) to obtain liver targeting of adenine arabinoside monophosphate for the treatment of chronic hepatitis B virus infection.
Selective delivery to the liver of antiviral nucleoside analogs coupled to a high molecular mass lactosaminated poly-l-lysine and administered to mice by intramuscular route
1995, Biochemical Pharmacology
In order to obtain hepatotropic conjugates of antiviral drugs suitable for intramuscular administration, three nucleoside analogs (adenine arabinoside monophosphate, ribavirin and azidothymidine) were coupled to a high molecular mass lactosaminated poly-l-lysine. The conjugates had a high molar ratio drug/conjugate and after intramuscular administration to mice, were selectively taken up by the liver and eliminated by the kidney only in minute quantities. The high molar ratio and low renal elimination are important properties not possessed by conjugates previously prepared by using a small molecular mass lactosaminated poly-l-lysine. The conjugate with adenine arabinoside monophosphate (ara-AMP) was found to be devoid of acute toxicity for mice and in spite of its high molecular dimension (M_n = ca. 72,500) did not induce antibodies in this animal after repeated intramuscular injections. This conjugate could have two advantages over a similar complex of ara-AMP with lactosaminated human albumin currently under clinical trials for the treatment of chronic type B hepatitis which must be injected intravenously: it might provide better patient compliance since it is injectable intramuscularly and could introduce larger amounts of ara-AMP into hepatocytes due to its higher drug/carrier molar ratio.
Ultralstructural and biochemical studies of the effect of polychlorinated biphenyl on mouse parotid gland cells
1995, Archives of Oral Biology
These effects of polychlorinated biphenyl (PCB) were examined by light and electron microscopy and biochemical analysis of lysosomal enzyme activities. Several experimental protocols with dosage schedules of either 0.2, 2.0, or 20 mg/kg of PCB were used. Typical histological changes were observed in mice given 2 mg/kg of PCB in a single injection. There were no remarkable changes until 4 days after PCB administration; marked cytoplasmic vacuolation was observed in parotid acinar cells at 7 days. The activities of lysosomal enzymes increased after the PCB injection and their maximum values appeared consistently at 4 days after the treatment; the increases were threefold for acid phosphatase, twofold for β-glucuronidase, threefold for cathepsin D, fivefold for cathepsin H and twofold for cathepsin L. As vacuolation was preceded by a large increase in lysosomal enzyme activities and the vacuoles co-localized with lysosomes, it is suggested that an increase in these activities induced by PCB may be closely related to the development of vacuolation in the parotid acinar cells as a subacute effect of PCB.

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Regular paperThe pathogenesis of vacuoles produced in rat and mouse liver cells by a conjugate of adenine arabinoside monophosphate with lactosaminated albumin

Abstract

Gastroenterology

FEBS Lett

Biochem Pharmacol

J Biol Chem

Lancet

Arch Biochem Biophys

Methods Cell Biol

J Biol Chem

J Biol Chem

J Biol Chem

Randomised controlled trial of adenine arabinoside 5′-monophosphate (ara-AMP) in chronic hepatitis B virus infection

Gut

Adenine arabinoside monophosphate (vidarabine phosphate) in combination with human leukocyte interferon in the treatment of chronic hepatitis B

Ann Intern Med

Randomised controlled trial of adenine arabinoside 5′-monophosphate in chronic active hepatitis B: comparison of the efficacy in heterosexual and homosexual patients

Hepatology

Regular paper
The pathogenesis of vacuoles produced in rat and mouse liver cells by a conjugate of adenine arabinoside monophosphate with lactosaminated albumin