S-Nitrosated human serum albumin dimer is not only a novel anti-tumor drug but also a potentiator for anti-tumor drugs with augmented EPR effects.

Macromolecules have been developed as carriers of low-molecular-weight drugs in drug delivery systems (DDS) to improve their pharmacokinetic profile or to promote their uptake in tumor tissue via enhanced permeability and retention (EPR) effects. In the present study, recombinant human serum albumin dimer (AL-Dimer), which was designed by linking two human serum albumin (HSA) molecules with the amino acid linker (GGGGS)(2), significantly accumulated in tumor tissue even more than HSA Monomer (AL-Monomer) and appearing to have good retention in circulating blood in murine colon 26 (C26) tumor-bearing mice. Moreover, we developed S-nitrosated AL-Dimer (SNO-AL-Dimer) as a novel DDS compound containing AL-Dimer as a carrier, and nitric oxide (NO) as (i) an anticancer therapeutic drug/cell death inducer and (ii) an enhancer of the EPR effect. We observed that SNO-AL-Dimer treatment induced apoptosis of C26 tumor cells in vitro, depending on the concentration of NO. In in vivo experiments, SNO-AL-Dimer was found to specifically deliver large amounts of cytotoxic NO into tumor tissue but not into normal organs in C26 tumor-bearing mice as compared with control (untreated tumor-bearing mice) and SNO-AL-Monomer-treated mice. Intriguingly, S-nitrosation improved the uptake of AL-Dimer in tumor tissue through augmenting the EPR effect. These data suggest that SNO-AL-Dimer behaves not only as an anticancer therapeutic drug, but also as a potentiator of the EPR effect. Therefore, SNO-AL-Dimer would be a very appealing carrier for utilization of the EPR effect in future development of cancer therapeutics.

[1]  P. Cullis,et al.  Drug Delivery Systems: Entering the Mainstream , 2004, Science.

[2]  H. Maeda,et al.  A new concept for macromolecular therapeutics in cancer chemotherapy: mechanism of tumoritropic accumulation of proteins and the antitumor agent smancs. , 1986, Cancer research.

[3]  H. Yasuda,et al.  Solid tumor physiology and hypoxia-induced chemo/radio-resistance: novel strategy for cancer therapy: nitric oxide donor as a therapeutic enhancer. , 2008, Nitric oxide : biology and chemistry.

[4]  Jeannette Bigler,et al.  Non-steroidal anti-inflammatory drugs for cancer prevention: promise, perils and pharmacogenetics , 2006, Nature Reviews Cancer.

[5]  W. Lu,et al.  Cationic albumin-conjugated pegylated nanoparticles allow gene delivery into brain tumors via intravenous administration. , 2006, Cancer research.

[6]  M. Dewhirst,et al.  Tumor vascular permeability, accumulation, and penetration of macromolecular drug carriers. , 2006, Journal of the National Cancer Institute.

[7]  Y. J. Tejwani,et al.  A new concept , 1984 .

[8]  Robert Langer,et al.  Advancing the field of drug delivery: taking aim at cancer. , 2003, Cancer cell.

[9]  S. Tanase,et al.  Recombinant Human Serum Albumin Dimer has High Blood Circulation Activity and Low Vascular Permeability in Comparison with Native Human Serum Albumin , 2006, Pharmaceutical Research.

[10]  J. Blohmer,et al.  A phase II study of topotecan plus gemcitabine in the treatment of patients with relapsed ovarian cancer after failure of first-line chemotherapy. , 2002, Annals of oncology : official journal of the European Society for Medical Oncology.

[11]  H. Terada,et al.  The triterpene celastrol as a very potent inhibitor of lipid peroxidation in mitochondria. , 1990, Biochemical and biophysical research communications.

[12]  H. Fiebig,et al.  Pre‐clinical evaluation of a methotrexate–albumin conjugate (MTX‐HSA) in human tumor xenografts in vivo , 2001, International journal of cancer.

[13]  Tetsuro Tanaka,et al.  Tumor targeting based on the effect of enhanced permeability and retention (EPR) and the mechanism of receptor-mediated endocytosis (RME). , 2004, International journal of pharmaceutics.

[14]  L. H. Reddy Drug delivery to tumours: recent strategies , 2005, The Journal of pharmacy and pharmacology.

[15]  Rakesh K Jain,et al.  Delivery of molecular and cellular medicine to solid tumors. , 1997, Advanced drug delivery reviews.

[16]  H. Maeda,et al.  Modulation of enhanced vascular permeability in tumors by a bradykinin antagonist, a cyclooxygenase inhibitor, and a nitric oxide scavenger. , 1998, Cancer research.

[17]  H. Maeda,et al.  Conjugates of Anticancer Agents and Polymers: Advantages of Macromolecular Therapeutics in vivo , 1993 .

[18]  M. Hashida,et al.  Pharmacokinetic analysis of in vivo disposition of succinylated proteins targeted to liver nonparenchymal cells via scavenger receptors: importance of molecular size and negative charge density for in vivo recognition by receptors. , 2002, The Journal of pharmacology and experimental therapeutics.

[19]  J. Wallace,et al.  The therapeutic potential of NO‐NSAIDs , 2003, Fundamental & clinical pharmacology.

[20]  B. Brüne,et al.  Nitric oxide-induced apoptosis: p53-dependent and p53-independent signalling pathways. , 1996, The Biochemical journal.

[21]  D. Kerr,et al.  Pharmacokinetic principles of locoregional chemotherapy. , 1993, Cancer surveys.

[22]  H. Maeda,et al.  Enhanced delivery of macromolecular antitumor drugs to tumors by nitroglycerin application , 2009, Cancer science.

[23]  Groopman Je Biologic and biochemical effects of mitoxantrone. , 1984 .

[24]  Felix Kratz,et al.  Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[25]  R. F. Chen,et al.  Removal of fatty acids from serum albumin by charcoal treatment. , 1967, The Journal of biological chemistry.

[26]  H. Maeda,et al.  Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[27]  Makoto Sato,et al.  Design and Evaluation of S-Nitrosylated Human Serum Albumin as a Novel Anticancer Drug , 2008, Journal of Pharmacology and Experimental Therapeutics.

[28]  H. Maeda,et al.  Tumoritropic and lymphotropic principles of macromolecular drugs. , 1989, Critical reviews in therapeutic drug carrier systems.

[29]  R. Jain Delivery of molecular and cellular medicine to solid tumors. , 2001, Advanced drug delivery reviews.

[30]  U. Bickel,et al.  Delivery of peptides and proteins through the blood-brain barrier. , 1993, Advanced drug delivery reviews.

[31]  G. Peters,et al.  In vitro and in vivo antitumor activity of methotrexate conjugated to human serum albumin in human cancer cells. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[32]  The determination of fracture mechanics properties of pharmaceutical materials in mode III loading using an anti-clastic plate bending method. , 2001, International journal of pharmaceutics.

[33]  T. Akaike,et al.  Cellular uptake mechanisms and responses to NO transferred from mono- and poly-S-nitrosated human serum albumin , 2011, Free radical research.

[34]  Min Zhang,et al.  Albumin Binding as a General Strategy for Improving the Pharmacokinetics of Proteins* , 2002, The Journal of Biological Chemistry.

[35]  J. Kutok,et al.  JS-K, a GST-activated nitric oxide generator, induces DNA double-strand breaks, activates DNA damage response pathways, and induces apoptosis in vitro and in vivo in human multiple myeloma cells. , 2007, Blood.

[36]  Y. Miyamoto,et al.  S-Nitrosylation of Human Variant Albumin Liprizzi (R410C) Confers Potent Antibacterial and Cytoprotective Properties , 2007, Journal of Pharmacology and Experimental Therapeutics.

[37]  N. Hogg Biological chemistry and clinical potential of S-nitrosothiols. , 2000, Free radical biology & medicine.

[38]  U. Bickel,et al.  Delivery of peptides and proteins through the blood-brain barrier. , 1993 .

[39]  T. Akaike Mechanisms of biological S-nitrosation and its measurement , 2000, Free radical research.

[40]  W. Hryniuk,et al.  Applications of dose intensity to problems in chemotherapy of breast and colorectal cancer. , 1987, Seminars in oncology.

[41]  H. Kubo,et al.  Randomized phase II trial comparing nitroglycerin plus vinorelbine and cisplatin with vinorelbine and cisplatin alone in previously untreated stage IIIB/IV non-small-cell lung cancer. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.