Synthesis and in Vitro Efficacy of MMP9-activated NanoDendrons

Overview

Journal Mol Pharm

Publisher American Chemical Society

Specialty Pharmacology

Date 2013 Jun 12

PMID 23750801

Citations 4

Authors

Lynn E Samuelson

Randy L Scherer

Lynn M Matrisian

J Oliver McIntyre

Darryl J Bornhop

Affiliations

Soon will be listed here.

Abstract

Chemotherapeutics such as doxorubicin (DOX) and paclitaxel (PXL) have dose-limiting systemic toxicities, including cardiotoxicity and peripheral neuropathy. Delivery strategies to minimize these undesirable effects are needed and could improve efficacy, while reducing patient morbidity. Here, DOX and PXL were conjugated to a nanodendron (ND) through an MMP9-cleavable peptide linker, producing two new therapies, ND2(DOX) and ND2(PXL), designed to improve delivery specificity to the tumor microenvironment and reduce systemic toxicity. Comparative cytotoxicity assays were performed between intact ND-drug conjugates and the MMP9 released drug in cell lines with and without MMP9 expression. While ND2(DOX) was found to lose cytotoxicity due to the modification of DOX for conjugation to the ND; ND2(PXL) was determined to have the desired properties for a prodrug delivery system. ND2(PXL) was found to be cytotoxic in MMP9-expressing mouse mammary carcinoma (R221A-luc) (53%) and human breast carcinoma (MDA-MB-231) (66%) at a concentration of 50 nM (in PXL) after 48 h. Treating ND2(PXL) with MMP9 prior to the cytotoxicity assay resulted in a faster response; however, both cleaved and intact versions of the drug reached the same efficacy as the unmodified drug by 96 h in the R221A-luc and MDA-MB-231 cell lines. Further studies in modified Lewis lung carcinoma cells that either do (LLC(MMP9)) or do not (LLC(RSV)) express MMP9 demonstrate the selectivity of ND2(PXL) for MMP9. LLC(MMP9) cells were only 20% viable after 48 h of treatment, while LLC(RSV) were not affected. Inclusion of an MMP inhibitor, GM6001, when treating the LLC(MMP9) cells with ND2(PXL) eliminated the response of the MMP9 expressing cells (LLC(MMP9)). The data presented here suggests that these NDs, specifically ND2(PXL), are nontoxic until activated by MMP9, a protease common in the microenvironment of tumors, indicating that incorporation of chemotherapeutic or cytostatic agents onto the ND platform have potential for tumor-targeted efficacy with reduced in vivo systemic toxicities.

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References

Bremer C, Tung C, Weissleder R . Molecular imaging of MMP expression and therapeutic MMP inhibition. Acad Radiol. 2002; 9 Suppl 2:S314-5. DOI: 10.1016/s1076-6332(03)80214-3. View

Scherer R, VanSaun M, McIntyre J, Matrisian L . Optical imaging of matrix metalloproteinase-7 activity in vivo using a proteolytic nanobeacon. Mol Imaging. 2009; 7(3):118-31. PMC: 2777890. View

Elsadek B, Kratz F . Impact of albumin on drug delivery--new applications on the horizon. J Control Release. 2011; 157(1):4-28. DOI: 10.1016/j.jconrel.2011.09.069. View

Jastrzebska B, Lebel R, Therriault H, McIntyre J, Escher E, Guerin B . New enzyme-activated solubility-switchable contrast agent for magnetic resonance imaging: from synthesis to in vivo imaging. J Med Chem. 2009; 52(6):1576-81. DOI: 10.1021/jm801411h. View

Kridel S, Chen E, Kotra L, Howard E, Mobashery S, Smith J . Substrate hydrolysis by matrix metalloproteinase-9. J Biol Chem. 2001; 276(23):20572-8. DOI: 10.1074/jbc.M100900200. View

McIntyre J, Fingleton B, Wells K, Piston D, Lynch C, Gautam S . Development of a novel fluorogenic proteolytic beacon for in vivo detection and imaging of tumour-associated matrix metalloproteinase-7 activity. Biochem J. 2003; 377(Pt 3):617-28. PMC: 1223892. DOI: 10.1042/BJ20030582. View

Achilefu S . Rapid response activatable molecular probes for intraoperative optical image-guided tumor resection. Hepatology. 2012; 56(3):1170-3. DOI: 10.1002/hep.25807. View

Wang J, Wheeler D, Zhang J, Achilefu S, Kang K . NIR fluorophore-hollow gold nanosphere complex for cancer enzyme-triggered detection and hyperthermia. Adv Exp Med Biol. 2012; 765:323-328. DOI: 10.1007/978-1-4614-4989-8_45. View

Martin M, Carter K, Jean-Philippe S, Chang M, Mobashery S, Thiolloy S . Effect of ablation or inhibition of stromal matrix metalloproteinase-9 on lung metastasis in a breast cancer model is dependent on genetic background. Cancer Res. 2008; 68(15):6251-9. PMC: 2789265. DOI: 10.1158/0008-5472.CAN-08-0537. View

10.

McCawley L, Matrisian L . Tumor progression: defining the soil round the tumor seed. Curr Biol. 2001; 11(1):R25-7. DOI: 10.1016/s0960-9822(00)00038-5. View

11.

Majoros I, Myc A, Thomas T, Mehta C, Baker Jr J . PAMAM dendrimer-based multifunctional conjugate for cancer therapy: synthesis, characterization, and functionality. Biomacromolecules. 2006; 7(2):572-9. DOI: 10.1021/bm0506142. View

12.

Kline T, Torgov M, Mendelsohn B, Cerveny C, Senter P . Novel antitumor prodrugs designed for activation by matrix metalloproteinases-2 and -9. Mol Pharm. 2005; 1(1):9-22. DOI: 10.1021/mp0340183. View

13.

Law B, Tung C . Proteolysis: a biological process adapted in drug delivery, therapy, and imaging. Bioconjug Chem. 2009; 20(9):1683-95. DOI: 10.1021/bc800500a. View

14.

Slamon D, Eiermann W, Robert N, Pienkowski T, Martin M, Press M . Adjuvant trastuzumab in HER2-positive breast cancer. N Engl J Med. 2011; 365(14):1273-83. PMC: 3268553. DOI: 10.1056/NEJMoa0910383. View

15.

McIntyre J, Scherer R, Matrisian L . Near-infrared optical proteolytic beacons for in vivo imaging of matrix metalloproteinase activity. Methods Mol Biol. 2010; 622:279-304. PMC: 4495730. DOI: 10.1007/978-1-60327-299-5_18. View

16.

Chau Y, Tan F, Langer R . Synthesis and characterization of dextran-peptide-methotrexate conjugates for tumor targeting via mediation by matrix metalloproteinase II and matrix metalloproteinase IX. Bioconjug Chem. 2004; 15(4):931-41. DOI: 10.1021/bc0499174. View

17.

Liotta L, Tryggvason K, Garbisa S, Hart I, FOLTZ C, Shafie S . Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature. 1980; 284(5751):67-8. DOI: 10.1038/284067a0. View

18.

Nasrolahi Shirazi A, Tiwari R, Chhikara B, Mandal D, Parang K . Design and biological evaluation of cell-penetrating peptide-doxorubicin conjugates as prodrugs. Mol Pharm. 2013; 10(2):488-99. DOI: 10.1021/mp3004034. View

19.

Albright C, Graciani N, Han W, Yue E, Stein R, Lai Z . Matrix metalloproteinase-activated doxorubicin prodrugs inhibit HT1080 xenograft growth better than doxorubicin with less toxicity. Mol Cancer Ther. 2005; 4(5):751-60. DOI: 10.1158/1535-7163.MCT-05-0006. View

20.

Scherer R, McIntyre J, Matrisian L . Imaging matrix metalloproteinases in cancer. Cancer Metastasis Rev. 2008; 27(4):679-90. DOI: 10.1007/s10555-008-9152-9. View