- Email: [email protected]
P102. Development of ultradeformable liposome entrapped with nitrosyl ruthenium complex for PDT application
Poster abstracts / Nitric Oxide 19 (2008) S43–S72 flow. Acute lung injury was induced by 40% total body surface area 3rd degree cutaneous burn and smoke inhalation (48 breaths of cotton smoke, <40 °C) under deep anesthesia. Study groups: (1) 7-NI: (0.01 mg/kg/h 7-NI, [2 ml/h]) was continuously infused into bronchial artery, n = 5. (2) Control: Same amount of saline was injected into bronchial artery, n = 5. After the injury, all animals were placed on ventilator, and fluid resuscitated equally. Experiment was conducted in awaken condition for 24 h. Results: Pulmonary function evaluated by measurement of blood gas analysis, pulmonary mechanics, and pulmonary transvascular fluid flux was severely deteriorated in control group. However, the above changes were significantly attenuated by 7-NI infusion (PaO2/FiO2 ratio; 7-NI:329 ± 43*, control:135 ± 25, lung lymph flow; 7-NI:13.9 ± 3.1, control:63.6 ± 10.0*, lung permeability index; 7-NI:7.0 ± 0.8, control:39.9 ± 4.5*, *p < 0.05 vs. control). Conclusions: Previously, we have reported beneficial effects of IV administered 7-NI with dose of 1 mg/kg/h in same model. In the present study we demonstrated that low (100 times lower) dose of 7-NI directly administered into bronchial artery attenuated the pulmonary dysfunction in same extent. Our data strongly suggest that local airway production of NO may increase airway blood flow and contributes to pulmonary dysfunction following smoke inhalation and burn. Regional delivery of the pharmaceutical compounds might be both beneficial and cost efficient in treatment of burn victims, especially those who have concomitant smoke inhalation. doi:10.1016/j.niox.2008.06.199
P102. Development of ultradeformable liposome entrapped with nitrosyl ruthenium complex for PDT application Renata Galvão de Lima a, Antonio Claudio Tedesco b, Roberto Santana da Silva c, Margareth Jayne Lawrence a a Pharmacy, King’s College of London b Chemistry, Universidade de São Paulo c Pharmacy, Universidade de São Paulo Photodynamic therapy (PDT) is based on the use of 1O2 (singlet oxygen), maybe this clinical therapy could be potentialized by production of reactive nitric oxide species (RNOs) like nitric oxide (NO). Recently, [Ru(NH.NHq)(tpy)NO]3+ (A) and ZnPc (B) compounds were loaded in liposome where were showed 1O2 and NO production under light stimulation (675 nm) [1]. Considering, the PDT is a modality for the treatment of superficial tumor, we proposed the development of ultradeformable liposome (UDL) loaded with A, B compounds and in association (A + B). UDL and non-UDL (conventional liposome) were prepared by the ethanolic injection method. Typically, an ethanolic solution which add phospholipids and surfactant (DOPC:DMPC:Tween 20) were injected with a syringe into PBS, under 40 °C and at a flow rate of 1 lL/s. Homogenously vesicle size were produced following extrusion through 100 and 50 nm polycarbonate filters using pressure filter holder under external pressure (1 bar), and characterized by dynamic light scattering. A, B and A + B were loaded in UDL and non-UDL resulting 100 and 50 nm mean vesicle size, respectively. Retention degree (RD) of A and B in vesicle preparations after passage through the filter barrier were determined by UV–visible, 91% and 95% of compounds remain in to preparation after filter 50 nm, respectively. Deformability index (DI) results indicated that empty-UDL and both A-UDL, B-UDL and A + B-UDL had the same elasticity (D = 3.0) and resulted 7-fold more elastic than non-UDL (D = 0.7). These results suggested that deformable liposomes might be of potential use for the formulation of nitrosyl ruthenium compounds in photodynamic treatment of skin diseases. Reference [1] D.S. Maranho, R.G. de Lima, R.S. da Silva, L.M. Bendhack, A.C. Tedesco, Photochemistry and Photobiology, submitted for publication. doi:10.1016/j.niox.2008.06.200
P103. Dinitrosyl iron complexes with thiolate ligands as future medicines Anatoly F. Vanin, Anatoly F. Vanin Russian Academy of Sciences, Semyonov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia Dinitrosyl-iron complexes (DNIC) with thiolate ligands (formula {(RS)2Fe+(NO+. . .0 SR)2}) are endogenous nitroso compounds which can be detected by EPR method in cells and tissues generating NO via L-arginine- or nitrite dependent pathways. Now water soluble dry powder preparation of DNIC with glutathione (DNIC-GS) was designed at industrial conditions which keeps its physiological and physico-chemical properties for a long time. Very low toxicity and no mutagenic or pro-apoptotic activities are characteristics of the preparation, that allows to consider
S67
it as a base for designing of a medicine with various properties. The following physiological effects of the preparation are considered in the report. 1. Powerful long-lasting hypotensive activity. Bolus i/v injections of the preparation solution into rats or monkeys at a dose of 2.5 or 1.0 lmol/kg DNIC-GS induces profound long-lasting average arterial pressure decrease followed by its gradual recovery to the initial level for 1 or 0.5 h, respectively. Further increase of the dose does not reinforce the effect. 2. High vasorelaxing activity. The preparation efficiently relaxes rat abdominal aorta rings with endothelium. Small tone restoration is observed during prolonged period registration (20–30 min) with the value of ED50 = 1.6 107 M DNIC-GS. The EPR assay of aliquots of the incubation medium supplemented with the preparation points to the high stability of the DNIC-GS in the medium at least 5–10 min that ensures evidently long-lasting vasodilatory action of the complex. 3. Capacity of blockade of pellet aggregation. The preparation at 0.2 mM concentration of DNIC-GS inhibits efficiently (90%) pellet aggregation in patient blood plasma induced by ADP or thrombin. 4. Acceleration of skin wound healing. Twofold injections of the preparation solution into rat wound tissue at total dose of 5 lmol DNIC-GS for 1 and 2 days after skin wounding results in granulocyte volume increase 3–4 times more than that for control wound. The average time of complete wound healing decreased to 17.5 ± 1.1 days versus 24.5 ± 1.6 days for control wounds. 5. Beneficial action on the state of cavernosum bodies of rat penis. Intracavernosum injection of the preparation into rat penis at the dose of 0.5 lmol induces prolonged (1–1.5 h) erection of the organ both in intact animals and in animals with denerveted penis. Also this treatment protects the denerveted cavernosum tissue against its transformation into fibrosis tissue.
doi:10.1016/j.niox.2008.06.201
P104. Photobiological effect of nitric oxide and singlet oxygen in cancer cells occasioned by light irradiation in the phototherapeutic window of a nitrosyl ruthenium conjugated to a phthalocyanine rare earth complex Simone Aparecida Cicillini a, Ana Carolina Prazias a, Antonio Cláudio Tedesco b, Osvaldo Antonio Serra b, Roberto Santana da Silva a a FCFRP—USP, Av do Café s/n, Ribeirão Preto-SP 14040-903, Brazil b Depto. de Química da FFCLRP-USP, Av. Bandeirantes 3900, Ribeirão Preto-SP 14040-901, Brazil The development of molecular systems that absorb light intensely and produces radical useful in clinical therapy is an important research goal in several different areas. One of these systems is phthalocyanines that have been used in clinical application as photodynamic therapy (PDT). We are presenting here the photochemical, photophysical and photobiological studies of a mixture containing cis-[Ru(dcbpy)2(Cl)(NO)] (dcbpy = 4,40 -dicarboxi-2,20 -bipyridine), (I), and Na4[Tb(TsPc)(acac)] (TsPc = tetrasulfonated phthalocyanines; acac = acetylacetone), (II) as a system capable to improve PDT. The specie (I) was obtained from [RuCl2(dcbpy)2] and (II) was obtained by reacting phthalocyanine with terbium acetylacetonate. The UV–vis spectrum of (I) presents bands in the region of 280 (p–p*) and 330 nm (MLCT) and of (II) presents the bands of phthalocyanines in 340 nm (Soret) and 680 nm (Q band). The FTIR spectrum of (I) shows mNO = 1930 cm1 that was characterized as nitrosyl ligand bonded on ruthenium center. The cyclic voltammogram of (I) complex showed two processes centered in the nitrosyl ligand, in aqueous solution: E = 0.10 V (NO+/0) and E = 0.50 V (NO0/) versus Ag/AgCl. The photolysis of (I/II) species is a fascination. The NO and 1O2 quantum yields for the light irradiation in 680 nm region were measured and shows values as [NO] = 1.21 ± 0.14 lmol L1 and øOS = 0.67. The photochemical pathway of (I/II) could be attributed to a photoinduced electron-transfer process. The cytotoxic assays of (I) and (I/II) carried out with B16F10 cells show decrease cell viability to 20% in the dark and 80% under light irradiation. Our results document that NO and 1O2 produced at the same time could improve PDT and maybe useful in cancer treatment. Acknowledgment Financial support: FAPESP and CNPq. doi:10.1016/j.niox.2008.06.202
P105. Sustained and controlled NO-releasing polymeric materials Dillip Mohanty, J. Wang, Gary Teng, H. Yu Department of Chemistry, Central Michigan University, Mt. Pleasant, MI 48858, USA
P102. Development of ultradeformable liposome entrapped with nitrosyl ruthenium complex for PDT application Renata Galvão de Lima a, Antonio Claudio Tedesco b, Roberto Santana da Silva c, Margareth Jayne Lawrence a a Pharmacy, King’s College of London b Chemistry, Universidade de São Paulo c Pharmacy, Universidade de São Paulo Photodynamic therapy (PDT) is based on the use of 1O2 (singlet oxygen), maybe this clinical therapy could be potentialized by production of reactive nitric oxide species (RNOs) like nitric oxide (NO). Recently, [Ru(NH.NHq)(tpy)NO]3+ (A) and ZnPc (B) compounds were loaded in liposome where were showed 1O2 and NO production under light stimulation (675 nm) [1]. Considering, the PDT is a modality for the treatment of superficial tumor, we proposed the development of ultradeformable liposome (UDL) loaded with A, B compounds and in association (A + B). UDL and non-UDL (conventional liposome) were prepared by the ethanolic injection method. Typically, an ethanolic solution which add phospholipids and surfactant (DOPC:DMPC:Tween 20) were injected with a syringe into PBS, under 40 °C and at a flow rate of 1 lL/s. Homogenously vesicle size were produced following extrusion through 100 and 50 nm polycarbonate filters using pressure filter holder under external pressure (1 bar), and characterized by dynamic light scattering. A, B and A + B were loaded in UDL and non-UDL resulting 100 and 50 nm mean vesicle size, respectively. Retention degree (RD) of A and B in vesicle preparations after passage through the filter barrier were determined by UV–visible, 91% and 95% of compounds remain in to preparation after filter 50 nm, respectively. Deformability index (DI) results indicated that empty-UDL and both A-UDL, B-UDL and A + B-UDL had the same elasticity (D = 3.0) and resulted 7-fold more elastic than non-UDL (D = 0.7). These results suggested that deformable liposomes might be of potential use for the formulation of nitrosyl ruthenium compounds in photodynamic treatment of skin diseases. Reference [1] D.S. Maranho, R.G. de Lima, R.S. da Silva, L.M. Bendhack, A.C. Tedesco, Photochemistry and Photobiology, submitted for publication. doi:10.1016/j.niox.2008.06.200
P103. Dinitrosyl iron complexes with thiolate ligands as future medicines Anatoly F. Vanin, Anatoly F. Vanin Russian Academy of Sciences, Semyonov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia Dinitrosyl-iron complexes (DNIC) with thiolate ligands (formula {(RS)2Fe+(NO+. . .0 SR)2}) are endogenous nitroso compounds which can be detected by EPR method in cells and tissues generating NO via L-arginine- or nitrite dependent pathways. Now water soluble dry powder preparation of DNIC with glutathione (DNIC-GS) was designed at industrial conditions which keeps its physiological and physico-chemical properties for a long time. Very low toxicity and no mutagenic or pro-apoptotic activities are characteristics of the preparation, that allows to consider
S67
it as a base for designing of a medicine with various properties. The following physiological effects of the preparation are considered in the report. 1. Powerful long-lasting hypotensive activity. Bolus i/v injections of the preparation solution into rats or monkeys at a dose of 2.5 or 1.0 lmol/kg DNIC-GS induces profound long-lasting average arterial pressure decrease followed by its gradual recovery to the initial level for 1 or 0.5 h, respectively. Further increase of the dose does not reinforce the effect. 2. High vasorelaxing activity. The preparation efficiently relaxes rat abdominal aorta rings with endothelium. Small tone restoration is observed during prolonged period registration (20–30 min) with the value of ED50 = 1.6 107 M DNIC-GS. The EPR assay of aliquots of the incubation medium supplemented with the preparation points to the high stability of the DNIC-GS in the medium at least 5–10 min that ensures evidently long-lasting vasodilatory action of the complex. 3. Capacity of blockade of pellet aggregation. The preparation at 0.2 mM concentration of DNIC-GS inhibits efficiently (90%) pellet aggregation in patient blood plasma induced by ADP or thrombin. 4. Acceleration of skin wound healing. Twofold injections of the preparation solution into rat wound tissue at total dose of 5 lmol DNIC-GS for 1 and 2 days after skin wounding results in granulocyte volume increase 3–4 times more than that for control wound. The average time of complete wound healing decreased to 17.5 ± 1.1 days versus 24.5 ± 1.6 days for control wounds. 5. Beneficial action on the state of cavernosum bodies of rat penis. Intracavernosum injection of the preparation into rat penis at the dose of 0.5 lmol induces prolonged (1–1.5 h) erection of the organ both in intact animals and in animals with denerveted penis. Also this treatment protects the denerveted cavernosum tissue against its transformation into fibrosis tissue.
doi:10.1016/j.niox.2008.06.201
P104. Photobiological effect of nitric oxide and singlet oxygen in cancer cells occasioned by light irradiation in the phototherapeutic window of a nitrosyl ruthenium conjugated to a phthalocyanine rare earth complex Simone Aparecida Cicillini a, Ana Carolina Prazias a, Antonio Cláudio Tedesco b, Osvaldo Antonio Serra b, Roberto Santana da Silva a a FCFRP—USP, Av do Café s/n, Ribeirão Preto-SP 14040-903, Brazil b Depto. de Química da FFCLRP-USP, Av. Bandeirantes 3900, Ribeirão Preto-SP 14040-901, Brazil The development of molecular systems that absorb light intensely and produces radical useful in clinical therapy is an important research goal in several different areas. One of these systems is phthalocyanines that have been used in clinical application as photodynamic therapy (PDT). We are presenting here the photochemical, photophysical and photobiological studies of a mixture containing cis-[Ru(dcbpy)2(Cl)(NO)] (dcbpy = 4,40 -dicarboxi-2,20 -bipyridine), (I), and Na4[Tb(TsPc)(acac)] (TsPc = tetrasulfonated phthalocyanines; acac = acetylacetone), (II) as a system capable to improve PDT. The specie (I) was obtained from [RuCl2(dcbpy)2] and (II) was obtained by reacting phthalocyanine with terbium acetylacetonate. The UV–vis spectrum of (I) presents bands in the region of 280 (p–p*) and 330 nm (MLCT) and of (II) presents the bands of phthalocyanines in 340 nm (Soret) and 680 nm (Q band). The FTIR spectrum of (I) shows mNO = 1930 cm1 that was characterized as nitrosyl ligand bonded on ruthenium center. The cyclic voltammogram of (I) complex showed two processes centered in the nitrosyl ligand, in aqueous solution: E = 0.10 V (NO+/0) and E = 0.50 V (NO0/) versus Ag/AgCl. The photolysis of (I/II) species is a fascination. The NO and 1O2 quantum yields for the light irradiation in 680 nm region were measured and shows values as [NO] = 1.21 ± 0.14 lmol L1 and øOS = 0.67. The photochemical pathway of (I/II) could be attributed to a photoinduced electron-transfer process. The cytotoxic assays of (I) and (I/II) carried out with B16F10 cells show decrease cell viability to 20% in the dark and 80% under light irradiation. Our results document that NO and 1O2 produced at the same time could improve PDT and maybe useful in cancer treatment. Acknowledgment Financial support: FAPESP and CNPq. doi:10.1016/j.niox.2008.06.202
P105. Sustained and controlled NO-releasing polymeric materials Dillip Mohanty, J. Wang, Gary Teng, H. Yu Department of Chemistry, Central Michigan University, Mt. Pleasant, MI 48858, USA