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Pharmacologic Studies of a Prodrug of Mitomycin C in Pegylated Liposomes (Promitil®): High Stability in Plasma and Rapid Thiolytic Prodrug Activation in Tissues

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Abstract

Purpose

Pegylated liposomal (PL) mitomycin C lipid-based prodrug (MLP) has recently entered clinical testing. We studied here the preclinical pharmacology of PL-MLP.

Methods

The stability, pharmacokinetics, biodistribution, and other pharmacologic parameters of PL-MLP were examined. Thiolytic cleavage of MLP and release of active mitomycin C (MMC) were studied using dithiothreitol (DTT), and by incubation with tissue homogenates.

Results

MLP was incorporated in the bilayer at 10% molar ratio with nearly 100% entrapment efficiency, resulting in a formulation with high plasma stability. In vitro, DTT induced cleavage of MLP with predictable kinetics, generating MMC and enhancing pharmacological activity. A long circulation half-life of MLP (10–15 h) was observed in rodents and minipigs. Free MMC is either extremely low or undetectable in plasma. However, urine from PL-MLP injected rats revealed delayed but significant excretion of MMC indicating in vivo activation of MLP. Studies in mice injected with H3-cholesterol radiolabeled PL-MLP demonstrated relatively greater tissue levels of H3-cholesterol than MLP. MLP levels were highest in tumor and spleen, and very low or undetectable in liver and lung. Rapid cleavage of MLP in various tissues, particularly in liver, was shown in ex-vivo experiments of PL-MLP with tissue homogenates. PL-MLP was less toxic in vivo than equivalent doses of MMC. Therapeutic studies in C26 mouse tumor models demonstrated dose-dependent improved efficacy of PL-MLP over MMC.

Conclusions

Thiolytic activation of PL-MLP occurs in tissues but not in plasma. Liposomal delivery of MLP confers a favorable pharmacological profile and greater therapeutic index than MMC.

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Abbreviations

Chol:

Cholesterol

DTT:

Dithiothreitol

HSPC:

Hydrogenated soybean phosphatidylcholine

IPA:

Isopropyl alcohol

MLP:

Mitomycin c lipid-based prodrug

MMC:

Mitomycin c

mPEG2000-DSPE:

methoxy-polyethylene glycol (2000)-phosphatidylethanolamine

MTD:

Maximal tolerated dose

PDI:

Polydispersity index

PK:

Pharmacokinetic(s)

PLD:

Pegylated liposomal doxorubicin

PL-MLP:

Pegylated liposomal mitomycin c lipid-based prodrug

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ACKNOWLEDGMENTS AND DISCLOSURES

Supported by the Israel Cancer Research Fund and by Lipomedix Pharmaceuticals Ltd. Special thanks to Sharon Wolf for TEM imaging at the Irving and Cherna Moskowitz Center for Nano and Bio-Nano Imaging of the Weizmann Institute of Science.

Alberto Gabizon is director and founder of Lipomedix Pharmaceuticals.

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Authors and Affiliations

  1. Lipomedix Pharmaceuticals Ltd., Giv’at Ram, Jerusalem, Israel

    Yasmine Amitay, Jenny Gorin & Patricia Ohana

  2. Shaare Zedek Medical Center, Jerusalem, Israel

    Hilary Shmeeda, Jenny Gorin, Dina Tzemach, Lidia Mak & Alberto Gabizon

  3. Hebrew University-School of Medicine, Jerusalem, Israel

    Yogita Patil & Alberto Gabizon

  4. Oncology Institute, Shaare Zedek MC, POB 3235, Jerusalem, 91031, Israel

    Alberto Gabizon

Authors
  1. Yasmine Amitay
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  2. Hilary Shmeeda
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  3. Yogita Patil
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  4. Jenny Gorin
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  5. Dina Tzemach
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  6. Lidia Mak
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  7. Patricia Ohana
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  8. Alberto Gabizon
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Correspondence to Alberto Gabizon.

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Amitay, Y., Shmeeda, H., Patil, Y. et al. Pharmacologic Studies of a Prodrug of Mitomycin C in Pegylated Liposomes (Promitil®): High Stability in Plasma and Rapid Thiolytic Prodrug Activation in Tissues. Pharm Res 33, 686–700 (2016). https://doi.org/10.1007/s11095-015-1819-7

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  • DOI: https://doi.org/10.1007/s11095-015-1819-7

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