Design of Cyclodextrin-derivatives at the edge between organic and supramolecular chemistry

Giornate di istituto ISOF 27 - 28 novembre 2023 Institute for Organic Synthesis and Photoreactivity Design of Cyclodextrin-derivatives @ the edge between organic & supramolecular chemistry Marco Agnes, PhD Istituto per la Sintesi Organica e la Fotoreattività (ISOF) Consiglio Nazionale delle Ricerche (CNR) Giornate di Istituto Bologna, 27-28 Novembre 2023 Giornate di istituto ISOF 27 - 28 novembre 2023 ERC Sectors Organic ♡ Supramolecular Chemistry 2(14) OH HO OH OH HO PE5_16 Supramolecular chemistry PE5_17 Organic chemistry LS7_3 Nanomedicine PE5 Synthetic Chemistry and Materials LS7 Prevention, Diagnosis and Treatment of Human Diseases 1 2 2014-2018: PhD @ NCSR Demokritos, Athens •Secondment: CycloLab, Budapest •Secondment: Hellenic Pasteur Institute, Athens 3 2020-2023: PostDoc @ ISOF-CNR Secondment: CarboHyde, Budapest 2023-2025: TD Researcher @ ISOF-CNR 2008-2014: BSc & MSc @ UniPV •Erasmus BSc: Utrecht Universiteit •Erasmus MSc: ICMAB-CSIC, Barcelona Giornate di istituto ISOF 27 - 28 novembre 2023 It is all about Cyclodextrins 3(14) n O OH OH HO O 1 23 5 4 6 Less acidic, most nucleophilic Most acidic, more nucleophilic More acidic, less nucleophilic Hydrophobic cavity Hydrophilic exterior H bond network n Label d (nm) 6 αCD 1,3 7 βCD 1,5 8 γCD 1,7 d Primary side (narrower) Secondary side (wider) Selectively tunable scaffold •Hydro-/Lipophilicity •Electrostatic properties Cyclodextrins •Macrocycles built of 1,4-α-glucopyranose units •Hydrophilic exterior + Hydrophobic cavity Pharmacological applications CD-based drug nanocarriers •Cavity dimension, flexibility, accessibilty Host:guest inclusion complexes •Enhanced water solubility and stability •Safe biocompatible profile Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 4(14) M. Agnes et al., Carbohydr. Polym. 2023, 121323 Inactived carbapenem MBL active site Carbapenems ◦Treatment of complex infections ◦Metallo-β-lactamase (MBL) enzymes: NO inhibitors available Meropenem Metal-Chelating CDs ◦Per-(6-iminodiacetic acid2,3-di-O-methyl) (IDAM)-CDs ◦Per-(6-iminodiacetic acid) (IDAM)-CDs Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 4(14) M. Agnes et al., Carbohydr. Polym. 2023, 121323 Metal-Chelating CDs ◦Per-(6-iminodiacetic acid2,3-di-O-methyl) (IDAM)-CDs ◦Per-(6-iminodiacetic acid) (IDAM)-CDs Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 4(15) M. Agnes et al., Carbohydr. Polym. 2023, 121323 Metal-Chelating CDs ◦Per-(6-iminodiacetic acid2,3-di-O-methyl) (IDAM)-CDs ◦Per-(6-iminodiacetic acid) (IDA)-CDs MALDI ToF (positive polarity) 1H NMR (500 MHz, 298 K, D2O, pH 6) Inverse-gated proton decoupled 13C NMR (63 MHz, 298 K, D2O, pH 6) = 7 BIDA 1 2 3 45 67 8 Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 5(14) pH 0 2 4 6 8 10 12 14  (ppm) 0 2 4 6 8 10 12 14 AIDAM BIDAM GIDAM BIDA pH 7 pKa1 pKa2 *Universidad de Valencia BIDAM M. Agnes et al., Carbohydr. Polym. 2023, 121323 13C NMR and Potentiometry*: pKa determination ◦Zwitterionic anionic at 4 < pH < 7: Physico-Chemical Properties Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 5(14) *Universidad de Valencia BIDAM BIDAM BIDA M. Agnes et al., Carbohydr. Polym. 2023, 121323 pH 0 2 4 6 8 10 12 14  (ppm) 0 2 4 6 8 10 12 14 AIDAM BIDAM GIDAM BIDA pKa1 pKa2 13C NMR and Potentiometry*: pKa determination ◦Zwitterionic anionic at 4 < pH < 7 Theoretical studies + 2D ROESY NMR: Cavity conformation → Guest inclusion ability ◦IDAM-CDs: flexible, distorted → Cavity NOT available ◦IDA-CDs: rigid, open → Cavity available Physico-Chemical Properties pH 7 Giornate di istituto ISOF 27 - 28 novembre 2023 Designed CD-derivatives to fight Antimicrobial Resistance (AMR) 5(14) Hc Hb Ha Hc Ha H5 H3 H5 H3 H6H7 Hb *Universidad de Valencia BIDAM BIDAM BIDA M. Agnes et al., Carbohydr. Polym. 2023, 121323 pH 0 2 4 6 8 10 12 14  (ppm) 0 2 4 6 8 10 12 14 AIDAM BIDAM GIDAM BIDA pKa1 pKa2 13C NMR and Potentiometry*: pKa determination ◦Zwitterionic anionic at 4 < pH < 7 Theoretical studies + 2D ROESY NMR: Cavity conformation → Guest inclusion ability ◦IDAM-CDs: flexible, distorted → Cavity NOT available ◦IDA-CDs: rigid, open → Cavity available Physico-Chemical Properties pH 7 Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy 8(14) Photochemical stepThermal steps [1] Angew. Chem. Int. Ed. 2016, 55, 3606 Ref. [1] 55% PABA ANBA NABA PNBA ◦Increased overall yields PABA: 12% → 41% Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy 8(14) Photochemical stepThermal steps [1] Angew. Chem. Int. Ed. 2016, 55, 3606 Ref. [1] 55% 94% ---% 84% ---% 015 30 45 60 0 20 40 60 80 100 PABA (375 nm) ANBA (368 nm) PNBA (305 nm) NABA (295 nm) A% Irradiation time (minutes) Substrate 1mM MB 50 mM DCM 9:1 THF Hg-577 nm 300 400 500 600 700 0.0 0.5 1.0 1.5 375 A l (nm) 0 0.5 1.0 1.5 2.0 3.0 4.0 5.0 7.5 10 15 20 30 45 60 0.1 cm Irradiation time (minutes) PABA PABA-O2 577 MB Hg-lamp 100W 577 nm filter DCM 9:1 THF 0.1 cm PABA ANBA NABA PNBA ◦Increased yield PABA-O2: 55% → 94% ◦Milder conditions ◦Increased overall yields PABA: 12% → 41% Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy M. Agnes et al., Chem. Eur. J. 2023, e202300511 9(14) OH HO OH OH HO PBS 1x pH 7.4 Max [O2] at a.p. DCM 9 THF 1 ≈ 2 mM ≈ 0.3 mM Photochemical Step in Water Solubilization of components in BCDPS Buffer aqueous solution (PBS 1x, pH 7.4) ◦MB : BCDPS → logK1:1 = 4.8 + logK1:2 = 2.1 ◦PABA, ANBA solubilized (mM) Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy 9(14) OH HO OH OH HO PBS 1x pH 7.4 Max [O2] at a.p. DCM 9 THF 1 ≈ 2 mM ≈ 0.3 mM Deuterated water | D2O PBS, pH 7.4 ◦No conversion of naphthalenes ◦Full conversion of anthracenes NON-Deuterated water | H2O PBS, pH 7.4 ◦(Slower) conversion + Full NMR characterization ◦Scale up (€ + quantitative extraction) Photochemical Step in Water 1O2 lifetime τDCM = 99 μs τD2O = 68 μs τH2O = 3μs M. Agnes et al., Chem. Eur. J. 2023, e202300511 Solubilization of components in BCDPS Buffer aqueous solution (PBS 1x, pH 7.4) ◦MB : BCDPS → logK1:1 = 4.8 + logK1:2 = 2.1 ◦PABA, ANBA solubilized (mM) Giornate di istituto ISOF 27 - 28 novembre 2023 ◦NMR + Abs + ESI-MS ◦Thermolysis (Δ): Heating (90 °C) ◦Photolysis (hν): Irradiation (265 nm) ◦PABA-O2 → PABA: ◦Controlled O2 release ◦Proof of concepts Exploiting CD-based polymers for anticancer therapy 9(14) OH HO OH OH HO PBS 1x pH 7.4 Max [O2] at a.p. DCM 9 THF 1 ≈ 2 mM ≈ 0.3 mM 300 325 350 375 400 425 450 0,00 0,05 0,10 0,15 0,20 0,25 A l (nm) 000 045 145 250 370 480 Heating time (minutes) PABA-O2 PABA Δ 0 2 4 6 8 0,00 0,05 0,10 0,15 0,20 0,25 A Time (h) PABA-O2 () PABA-O2 (IRR) ANBA-O2 () ANBA-O2 (IRR) DMSO Org. Solv. ≈ BCDPS in PBS PABA PABA-O2 0h 2h 4h 6h 8h 24h Ref Δ 1H NMR 500 MHz, DMSO-d6 Abs. spectra DMSO-d6 Deuterated water | D2O PBS, pH 7.4 ◦No conversion of naphthalenes ◦Full conversion of anthracenes NON-Deuterated water | H2O PBS, pH 7.4 ◦(Slower) conversion + Full NMR characterization ◦Scale up (€ + quantitative extraction) Photochemical Step in Water Cycloreversion M. Agnes et al., Chem. Eur. J. 2023, e202300511 Solubilization of components in BCDPS Buffer aqueous solution (PBS 1x, pH 7.4) ◦MB : BCDPS → logK1:1 = 4.8 + logK1:2 = 2.1 ◦PABA, ANBA solubilized (mM) √ X Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy 10(14) Simoultaneous co-encapsulation of 3 orthogonal therapeutic agents: ACHIEVED Chemotherapeutic Agent: PCX ◦BCDPS 10 mg/mL solubilize up to 400 μM Photosensitizer: Ce6 ◦1 : 1 → Kb ≈ 103 M-1 ◦2 : 1 → Kb ≈ 104 M-2 Oxygen Releasing Agents: ORAs ◦Co-encaspsulation: - 4 substrates PABA, ANBA, PNBA, NABA - 2 ORAs PABA-O2, ANBA-O2 - Dosage consistent PCX + Ce6 √ Giornate di istituto ISOF 27 - 28 novembre 2023 Exploiting CD-based polymers for anticancer therapy 10(14) 1200 1250 1300 1350 1400 -5000 0 5000 10000 15000 au l (nm) Ce6 Ce6 in EPI-pbCyD Ce6 + PCX + PABA in EPI-pbCyD Ce6 + PCX + PABA-O2 in EPI-pbCyD PDT 1270 nm emission of 1O2 (Deut. PBS, pH 7.4): - BCDPS 10 mg/mL - Ce6 50 μM - PCX 10 μM - ORAs 200 μM PCX 10 μM + Ce6 10 μM + ORAs 200 μM in BCDPS 10 mg/mL (PBS, 1cm) 300 400 500 600 700 0.0 0.5 1.0 1.5 2.0 2.5 3.0 PABA A l (nm) Arithmetic Experimental Arithmetic Experimental PABA_O2 650 675 700 725 750 775 800 0.0 5.0x104 1.0x105 1.5x105 2.0x105 2.5x105 3.0x105666 Counts l (nm) Ce6 Ce6 in EPI-pbCyD Ce6 in EPI-pbCyD + PCX + PABA Ce6 in EPI-pbCyD + PCX + PABA_O2 663 Normalized lexc = 640 nm Abs (+ CD): arith vs exp sums confirm co-loading Em (τ + λ): unaltered Ce6 excited state Ce6 1O2 production: PDT performs Simoultaneous co-encapsulation of 3 orthogonal therapeutic agents: ACHIEVED Chemotherapeutic Agent: PCX ◦BCDPS 10 mg/mL solubilize up to 400 μM Photosensitizer: Ce6 ◦1 : 1 → Kb ≈ 103 M-1 ◦2 : 1 → Kb ≈ 104 M-2 Oxygen Releasing Agents: ORAs ◦Co-encaspsulation: - 4 substrates PABA, ANBA, PNBA, NABA - 2 ORAs PABA-O2, ANBA-O2 - Dosage consistent PCX + Ce6 √ ◦Absorbance: Complete dissolution ◦Ce6 emission: same shift upon complexation ◦Ce6 excitation: NO static quenching ◦Ce6 lifetime (τ): NO dynamic quenching ◦1O2 emission: not quenched, PDT feasible Giornate di istituto ISOF 27 - 28 novembre 2023 CD-monomers and CD-polymers in pharmaceutical applications 11(14) HypoCyclo project ◦Optimized thermal & photochemical synthesis of ORAs (Photooxygenation + Cycloreversion in water) ◦Simultaneous co-loading of 3 drugs for novel combination cancer treatment CylonHit project ◦Designed synthesis of CD-monomers ◦Increase selectivity for host:guest interaction ◦Address AMR in current treatments ERC Sectors PE5_16 Supramolecular chemistry PE5_17 Organic chemistry LS7_3 Nanomedicine PE5 Synthetic Chemistry and Materials LS7 Prevention, Diagnosis and Treatment of Human Diseases Giornate di istituto ISOF 27 - 28 novembre 2023 CD-monomers and CD-polymers in pharmaceutical applications 11(14) HypoCyclo project ◦Optimized thermal & photochemical synthesis of ORAs (Photooxygenation + Cycloreversion in water) ◦Simultaneous co-loading of 3 drugs for novel combination cancer treatment ERC Sectors PE5_16 Supramolecular chemistry PE5_17 Organic chemistry LS7_3 Nanomedicine PE5 Synthetic Chemistry and Materials LS7 Prevention, Diagnosis and Treatment of Human Diseases CylonHit project ◦Designed synthesis of CD-monomers ◦Increase selectivity for host:guest interaction ◦Address AMR in current treatments Ongoing project ◦Photochemical step in flow ◦Computational design ORAs ◦Biocompatible conditions ◦Biological tests ◦Cytotoxicity, drugs + O2 release ◦PDT performance Matteo Calvaresi UniBO Paolo Dambruoso Daniele Veclani Giornate di istituto ISOF 27 - 28 novembre 2023 Design of CD-derivatives for other applications 12(14) Ecosystem for sustainable transition of Emilia Romagna Spoke 2: Clean energy production, storage and saving 2024-2025 calls ◦European Research Council (ERC) Starting Grant ◦Fondo Italiano Scienza (FIS) Starting Grant ◦CD-polymer decoration with PS ◦Water solubilization of hydrophobic antennas ◦CO2 encapsulation: Carbon Capture, Storage and Reuse ◦Electrochemical, Photochemical, PhotoElectrochemical Cells F. Tunioli et al., Chem. Eur. J. 2023, 29, e202301854 Wastewater treatment ◦Grafting of graphene with CD-monomers able to encapsulate PFAS Nicola Armaroli Manuela Melucci Andrea Barbieri Open to collaborate, brain storming, exchange of ideas