Small Molecules & Nanomaterials Serving Bio-medicine

Integrins are cell surface receptors playing essential roles in many physiological and pathological events. The relevance of RGD-dependent α_Vβ₃, α_Vβ₅, α_Vβ₆ and α₅β₁ integrins during tumour angiogenesis and tumour development is well established. This is also reflected by differential integrin expression patterns in normal versus tumour tissue as well as in different stages of tumour progression.

In recent years our efforts in the area of specific integrin ligands led to the development of a new series of g-aminocyclopentanecarboxylic acid (Acpca)-based or g-aminoproline (Amp)-based RGD-cyclotetrapeptides which turned out to be very potent α_Vβ₃, α_Vβ₅ and α₅β₁ integrin binders with IC₅₀ binding affinities in the nanomolar range. In particular, the covalent conjugation of our AmpRGD ligands with cytotoxic agents as well as their embodiment in liposomal nanoparticles were accomplished and biological properties of these bioconjugates were evaluated with encouraging results.

Focus of this research line is on the design, synthesis, and characterisation of novel and modulable integrin-targeted RGD-based pseudopeptide ligands and their conjugates for biomedical applications in the field of target-directed anti-cancer and anti-fibrosis therapy.

More recently, our research interest moved to the α₄β₁ integrin subfamily, which covers a prime role among the therapeutic targets of interest in fighting multiple sclerosis. The aim of our project is to design and synthesize a new class of small-molecule peptidomimetics able to effectively bind to the α₄β₁ receptor in order to interfere with the migration of lymphocytes T across the BBB with subsequent demyelinating effect.

Main publications and patents about this subject:

1. Andreucci L.; Bugatti K.; Peppicelli S.; Ruzzolini J.; Lulli M.; Calorini L.; Battistini L.; Zanardi F.; Sartori A. and Bianchini F. Nintedanib-αVβ6 Integrin Ligand Conjugates ReduceTGFβ-Induced EMT in Human Non-Small Cell Lung Cancer. Int. J. Mol. Sci. 2023, 24(2), 1475; https://doi.org/10.3390/ijms24021475

2. Bugatti, K.; Sartori, A.; Battistini, L.; Ruzzolini, J.; Nediani, C.; Curti, C.; Bianchini, F.; and Zanardi, F. Nintedanib-αVβ3 Integrin Ligand Dual-Targeting Conjugates towards Precision Treatment of Melanoma. EurJOC, 2022, e202200765, https://doi.org/10.1002/ejoc.202200765

3. Bugatti, K.; Andreucci, E.; Monaco, N.; Battistini, L.; Peppicelli, S.; Ruzzolini, J.; Curti, C.; Zanardi, F.; Bianchini, F. and Sartori, A. Nintedanib-Containing Dual Conjugates Targeting αVβ6 Integrin and Tyrosine Kinase Receptors as Potential Antifibrotic Agents. ACS Omega, 2022, 7, 21, 17658–17669, https://doi.org/10.1021/acsomega.2c00535

4. Sartori, A.; Bugatti, K.; Portioli, E.; Baiula, M.; Casamassima, I.; Bruno, A.; Bianchini, F.; Curti, C.; Zanardi, F. and Battistini, L. New 4-Aminoproline-Based Small Molecule Cyclopeptidomimetics as Potential Modulators of α4β1 Integrin. Molecules, 2021, 26(19), 6066. 10.3390/molecules26196066

5. Battistini, L.; Bugatti, K.; Sartori, A.; Curti, C.; Zanardi, F. RGD Peptide‐Drug Conjugates as Effective Dual Targeting Platforms: Recent Advances. Eur. J. Org. Chem. 2021, 2506 – 2528. doi.org/10.1002/ejoc.202100240

6. Bugatti, K.; Bruno, A.; Arosio, D.; Sartori, A.; Curti, C.; Augustijn, L.; Zanardi, F.; Battistini, L. Shifting Towards α_Vβ₆ Integrin Ligands Using Novel Aminoproline-Based Cyclic Peptidomimetics. Chem. Eur. J. 2020, 26, 13468-13475. doi.org/10.1002/chem.202002554

7. Sartori, A.; Corno, C.; De Cesare, M.; Scanziani, E.; Minoli, L.; Battistini, L.; Zanardi, F.; Perego, P. Efficacy of a Selective Binder of αVβ3 Integrin Linked to the Tyrosine Kinase Inhibitor Sunitinib in Ovarian Carcinoma Preclinical Models. Cancers, 2019, 11, 531. doi.org/10.3390/cancers11040531

8. Bianchini, F.; De Santis, A.; Portioli, E.; Russo Krauss, I.; Battistini, L.; Curti, C.; Peppicelli, S.; Calori, L.; D'Errico, G.; Zanardi, F.; Sartori, A.; Integrin-targeted AmpRGD sunitinib liposomes as integrated antiangiogenic tools, 2019, 18, 135-145. DOI: 10.1016/j.nano.2019.02.015

9. Bianchini, F.; Portioli, E.; Ferlenghi, F.; Vacondio, F.; Andreucci, E.; Biagioni, A.; Ruzzolini, J.; Peppicelli, S.; Lulli, M.; Calorini, L. Battistini, L.; Zanardi, F.; Sartori, A.; Cell-targeted c(AmpRGD)-sunitinib molecular conjugates impair tumor growth of melanoma. Cancer Lett. 2019, 446, 25-37. DOI: 10.1016/j.canlet.2018.12.021

10. Maggi, V.; Bianchini, F.; Portioli, E.; Peppicelli, S.; Lulli, M.; Bani, D.; Del Sole, R.; Zanardi, F.; Sartori, A.; Fiammengo, R. Gold Nanoparticles Functionalized with RGD-Semipeptides: A Simple yet Highly Effective Targeting System for αVβ3 Integrins. Chem. Eur. J. 2018, 24, 12093-12100. DOI: 10.1002/chem.201801823

11. Sartori, A.; Portioli, E.; Battistini, L.; Calorini, L.; Pupi, A.; Vacondio, F.; Arosio, D.; Bianchini, F.; Zanardi, F. Synthesis of Novel c(AmpRGD)-Sunitinib Dual Conjugates as Molecular Tools Targeting the α_Vβ₃ Integrin/VEGFR2 Couple and Impairing Tumor-Associated Angiogenesis. J. Med. Chem. 2017, 70(1), 248-262. doi: 10.1021/acs.jmedchem.6b01266

12. Battistini, L.; Burreddu, P.; Sartori, A.; Arosio, D.; Manzoni, L.; Paduano, L.; D’Errico, G.; Sala, R.; Reia, L.; Bonomini, S.; Rassu, G.; Zanardi, F. Enhancement of the Uptake and Cytotoxic Activity of Doxorubicin in Cancer Cells by Novel cRGD-Semipeptide-Anchoring Liposomes. Mol. Pharm. 2014, 11, 2280-2293. DOI: 10.1021/mp400718j

13. Pilkington-Miksa, M.; Arosio, D.; Battistini, L.; Belvisi, L.; De Matteo, M.; Vasile, F.; Burreddu, P.; Carta, P.; Rassu, G.; Perego, P.; Carenini, N.; Zunino, F.; De Cesare, M.; Castiglioni, V.; Scanziani, E.; Scolastico, C.; Casiraghi, G.; Zanardi, F.; Manzoni L. Design, Synthesis and Biological Evaluation of Novel cRGD-Paclitaxel Conjugates for Integrin-Assisted Drug Delivery. Bioconj. Chem. 2012, 23, 1610-1622. DOI: 10.1021/bc300164t

14. Auzzas, L.; Zanardi, F.; Battistini, L.; Burreddu, P.; Carta, P.; Rassu, G.;Curti, C.; Casiraghi, G. Targeting α_Vβ₃ Integrin: Design and Applications of Mono- and Multifunctional RGD-Based Peptides and Semipeptides. Curr. Med. Chem. 2010, 17, 1255-1299. DOI 10.2174/092986710790936301

15. Battistini, L.; Burreddu, P.; Carta, P.; Rassu, G.; Auzzas, L.; Curti, C.; Zanardi, F.; Manzoni, L.; Araldi, E. M. V.; Scolastico, C.; Casiraghi, G. 4-Aminoproline-Based Arginine-Glycine-Aspartate Integrin Binders with Exposed Ligation Points: Practical in-Solution Synthesis, Conjugation and Binding Affinity Evaluation. Org. Biomol. Chem. 2009, 7, 4924-4935. doi.org/10.1039/B914836A

16. Zanardi, F.; Burreddu, P.; Rassu, G.; Auzzas, L.; Battistini, L.; Curti, C.; Sartori, A.; Nicastro, G.; Menchi, G.; Cini, N.; Bottoncetti, A.; Raspanti, S.; Casiraghi, G. Discovery of Subnanomolar Arginine-Glycine-Aspartate-Based α_Vβ₃/α_Vβ₅ Integrin Binders Embedding 4-Aminoproline Residues. J. Med. Chem. 2008, 51, 1771-1782. doi.org/10.1021/jm701214z

17. Casiraghi, G.; Scolastico, C.; Zanardi, F.; Battistini, L.; Curti, C.; Rassu, G.; Auzzas, L.; Burreddu, P.; Manzoni, L. P. Integrin Targeted Cyclopeptide Ligands, Their Preparation and Use. EP N° 07003540.7, 21.02.2007; PCT Appl., N° PCT/IB2008/000366, 19.02.2008; USA Application Serial N° 12/527,603.

18. Casiraghi, G.; Rassu, G.; Auzzas, L.; Battistini, L.; Zanardi, F.; Curti, C.; Nicastro, G.; Belvisi, L.; Castorina, M.; Giannini, G.; Pisano, C. Ciclopeptidi Ibridi Contenenti Amminoacidi Cicloalcanici, e Loro Preparazione ed Uso. Ital. Pat. Appl. RE2005A00088, 21.07.05.

19. Casiraghi, G.; Rassu, G.; Auzzas, L.; Burreddu, P.; Gaetani, E.; Battistini, L.; Zanardi, F.; Curti, C.; Nicastro, G.; Belvisi, L.; Motto, I.; Castorina, M.; Giannini, G.; Pisano, C. Grafting Aminocyclopentane Carboxylic Acids onto the RGD Tripeptide Sequence Generates Low Nanomolar α_Vβ₃/α_Vβ₅ Integrin Dual Binders. J. Med. Chem. 2005, 48, 7675-7687.