Synthesis and characterization of ciprofloxacin imprinted polymer nanoparticles from renewable resources

In this study, novel molecularly imprinted polymer nanoparticles (MIPNPs) were synthesized from renewable resources. Maleinated castor oil (MACO) and lactic acid-maleic acid ester (LME), derived from renewable resources, were used as monomers in the synthesis. N,N′-methylenebis (acrylamide) (MBA) and TEMED/KPS (N,N,N,N-tetramethyl ethylene diamine/potassium persulfate were used as the cross-linker and the thermal initiator, respectively. Ciprofloxacin was used as the model drug molecule for the imprinting process. Polymerization was carried out at 50°C for 30, 60, and 90 minutes. Twelve different MIPs were synthesized, and the effects of the monomer types, initiator concentration, and duration of synthesis on the binding characteristics and particle sizes of MIPNPs were investigated. The produced polymeric materials were characterized by Fourier-transform infrared spectroscopy (FTIR) analysis. Furthermore, particle sizes of nanoparticles were measured using a Zetasizer particle size analyzer. Binding performances were investigated by UV-Vis spectroscopy. Finally, novel antibiotic imprinted polymer nanoparticles were obtained using renewable resources from plant oil-based materials. They can be used in many applications such as solid phase extraction, chromatography and drug delivery systems.

1. Synthesis and characterization of ciprofloxacin imprinted polymer nanoparticles from renewable resources Necla Yucel 1.2 , Elif Isikci Koca 1 , Hikmet Burcu Gencer 1 , Gokhan Cayli 3 , Pinar Cakir Hatir 1 1 Istanbul Arel University, Bioinspired Functional Polymers and Nanomaterials Laboratory (BIOFUNLAB) 2 Istanbul Medeniyet University, Nanoscience and Nanoengineering 3 Istanbul Cerrahpasa University, Faculty of Engineering, Department of Engineering Sciences [email protected] 1

2. Overview Molecularly imprinted polymer nanoparticles ( MIPnps ) Manufacturing of MIPnps Purpose of the study Experimental result Future work 2

3. 3

4. 4 Purpose of the Study To synthesize ciprofloxacin imprinted polymer nanoparticles from renewable resources. To reduce the dependence on petroleum resources and to minimize environmental harm .

5. Self- assembly CIPRO Lactic acid-Maleic acid ester (GC3) MBA Polymerization Remove template 5 Synthesis of MIPnps from GC3

6. Washed , centrifuged and dried in vacuum oven 1. All reagents were placed in vials . 2. 50 o C, 60min or 90 min 1. All reagent were placed in vials . 2. 50 o C, 30 min or 60 min. 3. Polymer nps were transferred into dialysis membrane 4. D.M. in washing solution 4. Lyophilisation MIP & NIP (gel forms ) MIPnps & NIPnps MIP & NIP 6 Synthesis Method

7. GC3 time ( min ) 1% 30 60 90 MIP - 22.34 57.99 NIP - 17.57 74.99 time (min) 2% 30 60 90 MIP 4.51 67.03 54.05 NIP 7.09 69.98 91.90 GC2 time (min) 1% 30 60 90 MIP - 76.97 58.66 NIP - 64.19 70.37 time ( min ) 2% 30 60 90 MIP 43.02 53.84 28.32 NIP 73.83 64.56 40.64 NIP GC2 GC3 initiator ratio (%) / time(min) 30 60 90 30 60 90 0.5 - - - - - - 1 - nano nano - nano gel 2 nano nano gel nano gel gel MIP GC2 GC3 initiator ratio (%) / time(min) 30 60 90 30 60 90 0.5 - - - - - - 1 - nano nano - nano gel 2 nano nano gel nano gel gel 7 Polymerization Yield %

8. Characterization of Ciprofloxacin Imprinted Polymer Nanoparticles Binding Capacity Loading Capacity and Release Profile Size Analysis ( Zeta-sizer ) Morphological Characterization (TEM) 8

9. Binding Capacity The largest difference between the binding of MIP and NIP was obtained with GC3 nanoparticles (1%, 60 min.). 9

10. Loading conditions : 0.5 mmol cipro sol. 24h- incubation , RT Centrifuged , discard supernatant Releasing conditions : PBS 1X RT UV- vis spect . w/ dialysis membrane 10 Release Profiles

11. Zeta-sizer Magnification - 6000x High voltage-100 kV TEM image 11 Particle Size of MIPnps Size d.nm % intensity GC3 %1, 30 min. MIPnp 189,9 58,5 100,8 39,7 30,18 1,7 NIPnp 189,9 58,5 100,8 39,7 30,18 1,7 %1, 60 min. MIPnp 178,4 97,6 40,1 2,4 NIPnp 188 100 %2, 30 min. MIPnp 186,1 100 NIPnp 147,4 66,7 69,62 33,3

12. Future work Binding capacity should be increased . Controlled release tests will be performed in different conditions . The synthesized polymer nanoparticles will be used in different applications depending on the intended use . Studies will be continued to obtain polymer nanoparticles with a more homogeneous distribution . 12

13. Assoc . Prof. Gokhan Cayli Dr. Pinar Cakir Hatir Dr. Elif Isıkci Koca Burcu Gencer Ehsan Sanattalab Necla Yucel Nur Muhammed Mustafaoğlu Seyma Turker 13 Funding EuroNanoMed III. project number 217S071-TUBITAK

14. References Altintas , Z. (2015), Molecular imprinting technology in advanced biosensors for diagnostics , Advances Biosensors Research , 1-30. Wackerlig , J. & Schirhagl , R. (2015), Applications of Molecularly Imprinted Polymer Nanoparticles and Their Advances toward Industrial Use : A Review , Analytical Chemistry , 88(1), 250–261. Canfarotta , F. Cecchini , A. & Piletsky . S. (2018), Nano-sized Molecularly Imprinted Polymers as Artificial Antibodies . Dong , J. Peng , Y. Gao , N. Bai , J. Ning , B. Liu , M. & Gao , Z. (2012), A novel polymerization of ultrathin sensitive imprinted film on surface plasmon resonance sensor, Analyst . 137(19), 4571-4576. Nicholls , I. A. Andersson , H. S. Golker , K. Henschel , H. Karlsson , B. C. Olsson , G. D. ... & Wikman , S. (2011), Rational design of biomimetic molecularly imprinted materials : theoretical and computational strategies for guiding nanoscale structured polymer development , Analytical and bioanalytical chemistry , 400(6), 1771. Bitar , M. Cayot , P. & Bou-Maroun , E. (2014), Molecularly imprinted polymer solid phase extraction of fungicides from wine samples, Analytical Methods, 6(16), 6467-6472. 14