8 th International Conference on Pharmaceuticals and Novel

8 th International Conference on Pharmaceuticals and Novel DD Systems NOVEL NON-RGD CYCLIC PEPTIDE DRUG CONJUGATES FOR TARGETED TREATMENT OF HUMAN METASTATIC MELANOMA Gary Gellerman Department of Chemical Sciences Ariel University Israel 1

Targeted Drug Delivery: present and future. ADC British Journal of Cancer (2008) NDC PDC

The Versatility of our Peptide-Platform-Drugs Conjugate Firer M. A. , Gellerman G. , J. Hematology & Oncology, (2012) Improvement of PK properties (BBB permeability, stability, solubility etc. )

Linkage Strategy Bioconjug Chem. (2010) ‘Traditional’ Linkers Ragozin et al. , Pep. Sci. (2015)

Diversity of the Peptidyl-based Platforms Highly tuneable

We aim to Ø Targeted Drug Delivery (TDD) of chemotherapeutic “cocktails” to reduce toxicity and increase efficacy Ø Reversing the drug resistance by TDD Ø Old drugs new opportunities

Multi drugs conjugates in the avb 3 integrin system

avb 3 integrin RGD biconjugates: Comparative Study (Cilengitide) Gilad Y. et al. , EJMC, (2014); Gilad Y. et al. , Bioorg. Med. Chem. (2015); Gilad Y. et al. , Pep. Sci (2016).

XTT Growth Inhibition Assay (2)

XTT Growth Inhibition Assay (3)

non-RGD αvβ 3 integrin specific peptide carrier ALOS 4 for targeted drug delivery in human metastatic melanoma Phage Display for αvβ 3 binding peptides Redko B. et al. submitted (2016)

ALOS 4 / αvβ 3 integrin docking simulations Two stretches of amino acids D 550 – T 553 (Thigh domain) and D 682 – L 693 (Calf-1 domain) show as sticks form the binding site (Figure 13 B) which is distinct from the RGD binding site.

Fluorescent microscopy experiment clearly shows the accumulation of ALOS 4 -FITC in WM-266 -4 human metastatic melanoma live cells. FITC ALOS 4 -FITC In vitro cytotoxicity on MMel cell lines vs HEK 293

ALOS 4 -FITC in Confocal Microscope in WM-266 -4 human metastatic melanoma cells 30 min A B B 1 h ALOS 4 -FITC

Ex Vivo fluorescence imaging studies on WM-266 -4 human metastatic melanoma nude mice model. Based on the results, the accumulation of the ALOS 4 -FITC was observed in the tumour tissue only and not in the spleen and liver.

Western blot

Conclusions and Future Plans • We developed HT loading methods for “cocktails” onto biodegradable peptidyl platforms • We established biostability and chemostability profiling • We have shown increased targeted cytotoxicity of peptide-drug “cocktail” conjugates vs peptide-mono drug counterparts • We developed novel non-RGD αvβ 3 integrin specific peptide carrier ALOS 4 for targeted drug delivery in human metastatic melanoma • In vivo study on other solid tumors

The TEAM: Chemistry: Tamara Brider Andrei Bazilevich Boris Redko Yossi Gilad Lena Regozin Alex Rozovsky Ariel Un. Biology: Prof. Albert Pinhasov Prof. Michael Firer Dr. Gabi Gerlitz Dr. Refael Minnes Helena Tuchinsky Ariel Un. Modeling: Dr. Dror Tobi Ariel Un. Thank You Grants: NOFAR Internal AU