TY  - JOUR
AU  - Ilić, Tanja
AU  - Đoković, Jelena
AU  - Nikolić, Ines
AU  - Mitrović, Jelena
AU  - Pantelić, Ivana
AU  - Savić, Snežana
AU  - Savić, Miroslav
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4515
AB  - Contemporary trends in combinatorial chemistry and the design of pharmaceuticals targeting brain disorders have favored the development of drug candidates with increased lipophilicity and poorer water solubility, with the expected improvement in delivery across the blood–brain barrier (BBB). The growing availability of innovative excipients/ligands allowing improved brain targeting and controlled drug release makes the lipid nanocarriers a reasonable choice to overcome the factors impeding drug delivery through the BBB. However, a wide variety of methods, study designs and experimental conditions utilized in the literature hinder their systematic comparison, and thus slows the advances in brain-targeting by lipid-based nanoparticles. This review provides an overview of the methods most commonly utilized during the preclinical testing of liposomes, nanoemulsions, solid lipid nanoparticles and nanostructured lipid carriers intended for the treatment of various CNS disorders via the parenteral route. In order to fully elucidate the structure, stability, safety profiles, biodistribution, metabolism, pharmacokinetics and immunological effects of such lipid-based nanoparticles, a transdisciplinary approach to preclinical characterization is mandatory, covering a comprehensive set of physical, chemical, in vitro and in vivo biological testing.
PB  - MDPI
T2  - Pharmaceutics
T1  - Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation
VL  - 15
IS  - 2
DO  - 10.3390/pharmaceutics15020443
ER  - 

@article{
author = "Ilić, Tanja and Đoković, Jelena and Nikolić, Ines and Mitrović, Jelena and Pantelić, Ivana and Savić, Snežana and Savić, Miroslav",
year = "2023",
abstract = "Contemporary trends in combinatorial chemistry and the design of pharmaceuticals targeting brain disorders have favored the development of drug candidates with increased lipophilicity and poorer water solubility, with the expected improvement in delivery across the blood–brain barrier (BBB). The growing availability of innovative excipients/ligands allowing improved brain targeting and controlled drug release makes the lipid nanocarriers a reasonable choice to overcome the factors impeding drug delivery through the BBB. However, a wide variety of methods, study designs and experimental conditions utilized in the literature hinder their systematic comparison, and thus slows the advances in brain-targeting by lipid-based nanoparticles. This review provides an overview of the methods most commonly utilized during the preclinical testing of liposomes, nanoemulsions, solid lipid nanoparticles and nanostructured lipid carriers intended for the treatment of various CNS disorders via the parenteral route. In order to fully elucidate the structure, stability, safety profiles, biodistribution, metabolism, pharmacokinetics and immunological effects of such lipid-based nanoparticles, a transdisciplinary approach to preclinical characterization is mandatory, covering a comprehensive set of physical, chemical, in vitro and in vivo biological testing.",
publisher = "MDPI",
journal = "Pharmaceutics",
title = "Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation",
volume = "15",
number = "2",
doi = "10.3390/pharmaceutics15020443"
}

Ilić, T., Đoković, J., Nikolić, I., Mitrović, J., Pantelić, I., Savić, S.,& Savić, M.. (2023). Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation. in Pharmaceutics
MDPI., 15(2).
https://doi.org/10.3390/pharmaceutics15020443

Ilić T, Đoković J, Nikolić I, Mitrović J, Pantelić I, Savić S, Savić M. Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation. in Pharmaceutics. 2023;15(2).
doi:10.3390/pharmaceutics15020443 .

Ilić, Tanja, Đoković, Jelena, Nikolić, Ines, Mitrović, Jelena, Pantelić, Ivana, Savić, Snežana, Savić, Miroslav, "Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation" in Pharmaceutics, 15, no. 2 (2023),
https://doi.org/10.3390/pharmaceutics15020443 . .

TY  - JOUR
AU  - Mitrović, Jelena
AU  - Divović-Matović, Branka
AU  - Knutson, Daniel E.
AU  - Petković, Miloš
AU  - Đorović, Đorđe
AU  - Ranđelović, Danijela V.
AU  - Dobričić, Vladimir
AU  - Đoković, Jelena
AU  - Lunter, Dominique J.
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4434
AB  - Preclinical development of deuterated pyrazoloquinolinone ligands, promising drug candidates for various neuropsychiatric disorders, was hindered by unusually low solubility in water and oils. DK-I-60-3 (7-methoxy-d3- 2-(4-methoxy-d3-phenyl)-2,5-dihydro-3Hpyrazolo[4,3-c]quinolin-3-one) is one of such pyrazoloquinolinones, and we recently reported about increased oral bioavailability of its nanocrystal formulation (NC). Lipid nano- particles (LNP) with a high concentration of lecithin, which enhances loading capacity of the lipid matrix, may give rise to further improvement. After preformulation studies by differential scanning calorimetry and polarized light microscopy, LNP were prepared by the hot high pressure homogenization, and characterized in terms of particle size, morphology, and encapsulation efficacy. The layered structure visible on atomic force micrographs was confirmed by nuclear magnetic resonance. Obtained formulations were desirably stable, with small particle size (<100 nm), and high encapsulation efficacy (>99 %). Lecithin was partially fluid and most probably located in the outer shell of the particle, together with DK-I-60-3. While the hydrophobic part of polysorbate 80 was completely immobilized, its hydrophilic part was free in the aqueous phase. In oral neuropharmacokinetic study in rats, an around 1.5-fold increase of area under the curve with LNP compared to NC was noticed both in brain and plasma. In bioavailability study, F value of LNP (34.7 ± 12.4 %) was 1.4-fold higher than of NC (24.5 ± 7.8 %); however, this difference did not reach statistical significance. Therefore, employment of LNP platform in preclinical formulation of DK-I-60-3 imparted an incremental improvement of its physicochemical as well as pharmacokinetic behavior.
PB  - Elsevier B.V.
T2  - International Journal of Pharmaceutics
T1  - High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances
VL  - 633
DO  - 10.1016/j.ijpharm.2023.122613
ER  - 

@article{
author = "Mitrović, Jelena and Divović-Matović, Branka and Knutson, Daniel E. and Petković, Miloš and Đorović, Đorđe and Ranđelović, Danijela V. and Dobričić, Vladimir and Đoković, Jelena and Lunter, Dominique J. and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Preclinical development of deuterated pyrazoloquinolinone ligands, promising drug candidates for various neuropsychiatric disorders, was hindered by unusually low solubility in water and oils. DK-I-60-3 (7-methoxy-d3- 2-(4-methoxy-d3-phenyl)-2,5-dihydro-3Hpyrazolo[4,3-c]quinolin-3-one) is one of such pyrazoloquinolinones, and we recently reported about increased oral bioavailability of its nanocrystal formulation (NC). Lipid nano- particles (LNP) with a high concentration of lecithin, which enhances loading capacity of the lipid matrix, may give rise to further improvement. After preformulation studies by differential scanning calorimetry and polarized light microscopy, LNP were prepared by the hot high pressure homogenization, and characterized in terms of particle size, morphology, and encapsulation efficacy. The layered structure visible on atomic force micrographs was confirmed by nuclear magnetic resonance. Obtained formulations were desirably stable, with small particle size (<100 nm), and high encapsulation efficacy (>99 %). Lecithin was partially fluid and most probably located in the outer shell of the particle, together with DK-I-60-3. While the hydrophobic part of polysorbate 80 was completely immobilized, its hydrophilic part was free in the aqueous phase. In oral neuropharmacokinetic study in rats, an around 1.5-fold increase of area under the curve with LNP compared to NC was noticed both in brain and plasma. In bioavailability study, F value of LNP (34.7 ± 12.4 %) was 1.4-fold higher than of NC (24.5 ± 7.8 %); however, this difference did not reach statistical significance. Therefore, employment of LNP platform in preclinical formulation of DK-I-60-3 imparted an incremental improvement of its physicochemical as well as pharmacokinetic behavior.",
publisher = "Elsevier B.V.",
journal = "International Journal of Pharmaceutics",
title = "High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances",
volume = "633",
doi = "10.1016/j.ijpharm.2023.122613"
}

Mitrović, J., Divović-Matović, B., Knutson, D. E., Petković, M., Đorović, Đ., Ranđelović, D. V., Dobričić, V., Đoković, J., Lunter, D. J., Cook, J. M., Savić, M.,& Savić, S.. (2023). High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances. in International Journal of Pharmaceutics
Elsevier B.V.., 633.
https://doi.org/10.1016/j.ijpharm.2023.122613

Mitrović J, Divović-Matović B, Knutson DE, Petković M, Đorović Đ, Ranđelović DV, Dobričić V, Đoković J, Lunter DJ, Cook JM, Savić M, Savić S. High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances. in International Journal of Pharmaceutics. 2023;633.
doi:10.1016/j.ijpharm.2023.122613 .

Mitrović, Jelena, Divović-Matović, Branka, Knutson, Daniel E., Petković, Miloš, Đorović, Đorđe, Ranđelović, Danijela V., Dobričić, Vladimir, Đoković, Jelena, Lunter, Dominique J., Cook, James M., Savić, Miroslav, Savić, Snežana, "High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances" in International Journal of Pharmaceutics, 633 (2023),
https://doi.org/10.1016/j.ijpharm.2023.122613 . .

TY  - CONF
AU  - Savić, Miroslav
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5006
AB  - In the 21st century, the pace of development of new pharmacological treatments for general medical and psychiatric disorders is remarkably different. While successes for the former are frequent, new drugs for mental, behavioral and neurodevelopmental disorders are sporadic. Many factors contribute to this discrepancy. Probably the most important is the complexity of etiology and manifestations of mood, psychotic, anxiety, neurocognitive and other disorders that are commonly treated with psycho pharmacological drugs. Although this factor cannot be directly addressed in drug development, optimization of drug exposure in brain tissue can certainly help to balance the efficacy and safety of both widely used and novel psycho pharmacological drugs. While drug exposure of various organs and tissues can be easily assessed from free (unbound) blood concentrations, the central nervous system (CNS) has a number of barriers, most notably the blood-brain barrier (BBB) that separates nervous tissue from the periphery. Optimised CNS exposure of a drug to its target site over a desired time period is critical to triggering its therapeutic effect. The presence of the BBB readily leads to an asymmetry of drug (unbound) exposure in the brain and in the systemic circulation, which prohibits the use of unbound drug concentration in plasma as a surrogate for unbound drug concentration in the brain. Comprehensive pharmacokinetic/pharmacodynamic studies of marketed CNS drugs have shown that the concentration of unbound drug in brain interstitial fluid is an appropriate measure of CNS exposure in the context of cell membrane targets of action. The more recent concept of brain-plasma partition coefficient, Kp,uu, as a parameter describing the relationship between the concentration of unbound drug in brain interstitial fluid and the concentration of unbound drug in plasma, is assumed to be the most important means of assessing brain exposure. Experimental assessment of Kp,uu requires either measurement of the brain unbound drug concentration in the interstitial fluid by microdialysis in vivo or estimation of the fraction of unbound drug in the whole brain homogenate (fu,brain) by equilibrium dialysis in vitro. Kp,uu can be calculated by dividing either the area under the curve (AUC) of the profile of the concentration of unbound drug in brain and plasma after a single administration or the steady-state unbound concentrations of drug in brain interstitial fluid and plasma. Although many marketed psychotropic drugs have Kp,uu values as low as 0.1-0.2, it is theorized that the most successful small molecule CNS drugs should have Kp,uu values near 1. The neuropharmacokinetic behavior of benzodiazepine compounds, both widely used and those in preclinical and clinical development, is presented as a showcase. Given the variety of receptor subpopulations at which these drugs act as positive allosteric modulators of GABAA receptors, the complexity of linking brain exposure data to the corresponding pharmacodynamic effect is explained.
PB  - International Association of Physical Chemists
C3  - 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
T1  - Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs
SP  - 6
EP  - 6
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5006
ER  - 

@conference{
author = "Savić, Miroslav",
year = "2023",
abstract = "In the 21st century, the pace of development of new pharmacological treatments for general medical and psychiatric disorders is remarkably different. While successes for the former are frequent, new drugs for mental, behavioral and neurodevelopmental disorders are sporadic. Many factors contribute to this discrepancy. Probably the most important is the complexity of etiology and manifestations of mood, psychotic, anxiety, neurocognitive and other disorders that are commonly treated with psycho pharmacological drugs. Although this factor cannot be directly addressed in drug development, optimization of drug exposure in brain tissue can certainly help to balance the efficacy and safety of both widely used and novel psycho pharmacological drugs. While drug exposure of various organs and tissues can be easily assessed from free (unbound) blood concentrations, the central nervous system (CNS) has a number of barriers, most notably the blood-brain barrier (BBB) that separates nervous tissue from the periphery. Optimised CNS exposure of a drug to its target site over a desired time period is critical to triggering its therapeutic effect. The presence of the BBB readily leads to an asymmetry of drug (unbound) exposure in the brain and in the systemic circulation, which prohibits the use of unbound drug concentration in plasma as a surrogate for unbound drug concentration in the brain. Comprehensive pharmacokinetic/pharmacodynamic studies of marketed CNS drugs have shown that the concentration of unbound drug in brain interstitial fluid is an appropriate measure of CNS exposure in the context of cell membrane targets of action. The more recent concept of brain-plasma partition coefficient, Kp,uu, as a parameter describing the relationship between the concentration of unbound drug in brain interstitial fluid and the concentration of unbound drug in plasma, is assumed to be the most important means of assessing brain exposure. Experimental assessment of Kp,uu requires either measurement of the brain unbound drug concentration in the interstitial fluid by microdialysis in vivo or estimation of the fraction of unbound drug in the whole brain homogenate (fu,brain) by equilibrium dialysis in vitro. Kp,uu can be calculated by dividing either the area under the curve (AUC) of the profile of the concentration of unbound drug in brain and plasma after a single administration or the steady-state unbound concentrations of drug in brain interstitial fluid and plasma. Although many marketed psychotropic drugs have Kp,uu values as low as 0.1-0.2, it is theorized that the most successful small molecule CNS drugs should have Kp,uu values near 1. The neuropharmacokinetic behavior of benzodiazepine compounds, both widely used and those in preclinical and clinical development, is presented as a showcase. Given the variety of receptor subpopulations at which these drugs act as positive allosteric modulators of GABAA receptors, the complexity of linking brain exposure data to the corresponding pharmacodynamic effect is explained.",
publisher = "International Association of Physical Chemists",
journal = "10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6",
title = "Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs",
pages = "6-6",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5006"
}

Savić, M.. (2023). Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
International Association of Physical Chemists., 6-6.
https://hdl.handle.net/21.15107/rcub_farfar_5006

Savić M. Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6. 2023;:6-6.
https://hdl.handle.net/21.15107/rcub_farfar_5006 .

Savić, Miroslav, "Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs" in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6 (2023):6-6,
https://hdl.handle.net/21.15107/rcub_farfar_5006 .

TY  - CONF
AU  - Stanković, Tijana
AU  - Ilić, Tanja
AU  - Pantelić, Ivana
AU  - Tošić, Anđela
AU  - Mitrović, Jelena
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5000
AB  - Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors 

Tijana Stanković1, Tanja Ilić1, Ivana Pantelić1, Anđela Tošić1, Jelena Mitrović1, James M. Cook2, Miroslav Savić3, Snežana Savić1

1 University of Belgrade-Faculty of Pharmacy, Department of Pharmaceutical Technology and Cosmetology, Vojvode Stepe 450, Belgrade, Serbia,
2 University of Wisconsin-Milwaukee, Milwaukee Institute for Drug Discovery, 3210 N. Cramer St. Milwaukee, Wisconsin, United States,
3 University of Belgrade-Faculty of Pharmacy, Department of Pharmacology, Vojvode Stepe 450, Belgrade, Serbia.

The poor water solubility of novel patent-protected ligand of the pyrazoloquinolinone chemotype (CW-02-79), with significant binding affinity for sigma-2 receptors in the brain, restricts the development of conventional parenteral formulations and consequently, extensive pharmacological studies during the preclinical investigation. Therefore, we aimed to develop a biocompatible nanocarrier tailored to specific physicochemical properties of CW-02-79, to improve its transport across the blood-brain barrier and achieve the optimal brain disposition. In this context, a detailed analysis of lipophilicity (via log P and log D determination), solubility in various solvents/excipients (using shake-flask method) and crystalline state of CW-02-07 (using X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC) with melt quenching approach and polarization microsocopy) was performed. After the analysis of key “input” physicochemical descriptors, based on the developed decision tree, nanoemulsions were selected as promising carriers for CW-02-79. The nanoemulsions were prepared using the high pressure homogenization method, varying the process (number of cycles, temperature and pressure) and formulation parameters (the content of the oil phase, the stabilizer mixture composition). Additionally, the influence of the sterilization process (thermal sterilization/aseptic filtration) on the nanoemulsion physicochemical properties was investigated, including droplet size and size distribution, zeta potential, pH, electrical conductivity and osmolality. The obtained results showed that it was possible to formulate CW-02-79-loaded nanoemulsions with 20% oil phase (medium chain triglycerides:castor oil at ratio 1:1), stabilized with the biocompatible emulsifiers (lecithin/polysorbate 80), exhibiting the nano-sized droplets (<200 nm) with narrow size distribution (polydispersity index < 0.2), zeta potential (> ǀ-30ǀ mV), pH (~ 5.7) and osmolality (295 mOsm/kg). The sterilization process did not remarkably affect the physiochemical properties of nanoemulsions, making them suitable for the parenteral administration. Owing to sastifying solubilization capacity for CW-02-79, physicochemical properties and preliminary stability, the nanoemulsions are the promising carriers worth exploring further to support the preclinical evalution of CW-02-79.
ACKNOWLEDGEMENT. This research was supported by the Science Fund of the Republic of Serbia, Grant No. 7749108, Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platform — NanoCellEmoCog
PB  - International Association of Physical Chemists
C3  - 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
T1  - Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5000
ER  - 

@conference{
author = "Stanković, Tijana and Ilić, Tanja and Pantelić, Ivana and Tošić, Anđela and Mitrović, Jelena and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors 

Tijana Stanković1, Tanja Ilić1, Ivana Pantelić1, Anđela Tošić1, Jelena Mitrović1, James M. Cook2, Miroslav Savić3, Snežana Savić1

1 University of Belgrade-Faculty of Pharmacy, Department of Pharmaceutical Technology and Cosmetology, Vojvode Stepe 450, Belgrade, Serbia,
2 University of Wisconsin-Milwaukee, Milwaukee Institute for Drug Discovery, 3210 N. Cramer St. Milwaukee, Wisconsin, United States,
3 University of Belgrade-Faculty of Pharmacy, Department of Pharmacology, Vojvode Stepe 450, Belgrade, Serbia.

The poor water solubility of novel patent-protected ligand of the pyrazoloquinolinone chemotype (CW-02-79), with significant binding affinity for sigma-2 receptors in the brain, restricts the development of conventional parenteral formulations and consequently, extensive pharmacological studies during the preclinical investigation. Therefore, we aimed to develop a biocompatible nanocarrier tailored to specific physicochemical properties of CW-02-79, to improve its transport across the blood-brain barrier and achieve the optimal brain disposition. In this context, a detailed analysis of lipophilicity (via log P and log D determination), solubility in various solvents/excipients (using shake-flask method) and crystalline state of CW-02-07 (using X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC) with melt quenching approach and polarization microsocopy) was performed. After the analysis of key “input” physicochemical descriptors, based on the developed decision tree, nanoemulsions were selected as promising carriers for CW-02-79. The nanoemulsions were prepared using the high pressure homogenization method, varying the process (number of cycles, temperature and pressure) and formulation parameters (the content of the oil phase, the stabilizer mixture composition). Additionally, the influence of the sterilization process (thermal sterilization/aseptic filtration) on the nanoemulsion physicochemical properties was investigated, including droplet size and size distribution, zeta potential, pH, electrical conductivity and osmolality. The obtained results showed that it was possible to formulate CW-02-79-loaded nanoemulsions with 20% oil phase (medium chain triglycerides:castor oil at ratio 1:1), stabilized with the biocompatible emulsifiers (lecithin/polysorbate 80), exhibiting the nano-sized droplets (<200 nm) with narrow size distribution (polydispersity index < 0.2), zeta potential (> ǀ-30ǀ mV), pH (~ 5.7) and osmolality (295 mOsm/kg). The sterilization process did not remarkably affect the physiochemical properties of nanoemulsions, making them suitable for the parenteral administration. Owing to sastifying solubilization capacity for CW-02-79, physicochemical properties and preliminary stability, the nanoemulsions are the promising carriers worth exploring further to support the preclinical evalution of CW-02-79.
ACKNOWLEDGEMENT. This research was supported by the Science Fund of the Republic of Serbia, Grant No. 7749108, Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platform — NanoCellEmoCog",
publisher = "International Association of Physical Chemists",
journal = "10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6",
title = "Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5000"
}

Stanković, T., Ilić, T., Pantelić, I., Tošić, A., Mitrović, J., Cook, J. M., Savić, M.,& Savić, S.. (2023). Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
International Association of Physical Chemists..
https://hdl.handle.net/21.15107/rcub_farfar_5000

Stanković T, Ilić T, Pantelić I, Tošić A, Mitrović J, Cook JM, Savić M, Savić S. Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5000 .

Stanković, Tijana, Ilić, Tanja, Pantelić, Ivana, Tošić, Anđela, Mitrović, Jelena, Cook, James M., Savić, Miroslav, Savić, Snežana, "Design of tailor-made biocompatible nanocarrier for novel pyrazoloquinolinone ligand (CW-02-79) based on comprehensive evaluation of critical physicochemical descriptors" in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6 (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5000 .

TY  - CONF
AU  - Đoković, Jelena
AU  - Marković, Bojan
AU  - Sharmin, Dishary
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4999
AB  - Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application
Jelena Đoković1, Bojan Marković2, Dishary Sharmin3, James M Cook3, Miroslav Savić4, Snežana Savić1
1University of pharmacy - Faculty of pharmacy, Department of pharmaceutical technology and cosmetology, 11221 Belgrade, Serbia
2University of pharmacy - Faculty of pharmacy, Department of pharmaceutical chemistry, 11221 Belgrade, Serbia
3Department of Chemistry and Biochemistry and the Milwaukee Institute for Drug Discovery, University of Wisconsin-Milwaukee, Milwaukee, WI, 53201, USA
4University of pharmacy - Faculty of pharmacy, Department of pharmacology, 11221 Belgrade, Serbia
The maximum amount of drug that can be incorporated into lipid nanoemulsions (NE) is usually judged by their solubility in the internal phase of the formulation. This can lead to various problems, such as precipitation of the drug after processing the formulation or, depending on the preparation technique used, the use of a large amount of the drug. To this end, it is useful to consider other drug loading methods, especially in the early stages of formulation development. In this study, we aimed to find the best way to achieve the highest loading of GL-II -73 in NEs for future parenteral applications for in vivo animal studies. This ligand acts as a positive allosteric modulator at α-GABAA receptors with combined antidepressant and cognition enhancing effects. NEs were prepared using the high pressure homogenization technique, a standard technique for parenteral NE preparation. The oil phase (medium-chain triglycerides, soy lecithin, and butylated hydroxytoluene) and the aqueous phase (glycerol, polysorbate 80, and 0.01 M phosphate buffer, pH 8) were separately heated to 50 ˚C and mixed until all components were dissolved. The aqueous phase was added to the oil phase and processed first on a rotor-stator homogenizer at 11000 rpm for 1 minute and then on a high-pressure homogenizer at 800 bar for 10 cycles. This resulted in a droplet size of 117.1 ± 1.5 nm, a PDI of 0.060 ± 0.008, a zeta potential of - 43.3 ± 1.3 mV, a pH of 7.89 ± 0.02, and a conductivity of 1061.67 ± 5.51 S/cm, indicating initial suitability for parenteral use. Using the empirical method for drug loading, we were able to dissolve GL-II -73 in the oil phase and achieve a drug concentration of 1.5 mg/ml in NE. For our experimental setup, this required the use of 120 mg of GL-II -73 per experiment. For the passive drug loading procedure, we incubated 1 ml of the placebo NE in the eppendorf tube (in duplicate) with the excess drug (approximately 10 mg per tube) for 72 h. The drug was then added to the eppendorf tube. The excess of the undissolved drug was removed after centrifugation. The drug content in the supernatant was 3.10 ± 0.25 mg/ml, indicating above-average loading of the drug and possibly suggesting localization of the drug in the droplet-stabilising layer, but this needs to be further demonstrated. This approach could contribute to more rational formulation development in the selection of formulation factors.
Acknowledgments
This research was funded by the Science Fund of the Republic of Serbia, GRANT No 7749108, Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platform - NanoCellEmоCog.
PB  - International Association of Physical Chemists
C3  - 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
T1  - Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4999
ER  - 

@conference{
author = "Đoković, Jelena and Marković, Bojan and Sharmin, Dishary and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application
Jelena Đoković1, Bojan Marković2, Dishary Sharmin3, James M Cook3, Miroslav Savić4, Snežana Savić1
1University of pharmacy - Faculty of pharmacy, Department of pharmaceutical technology and cosmetology, 11221 Belgrade, Serbia
2University of pharmacy - Faculty of pharmacy, Department of pharmaceutical chemistry, 11221 Belgrade, Serbia
3Department of Chemistry and Biochemistry and the Milwaukee Institute for Drug Discovery, University of Wisconsin-Milwaukee, Milwaukee, WI, 53201, USA
4University of pharmacy - Faculty of pharmacy, Department of pharmacology, 11221 Belgrade, Serbia
The maximum amount of drug that can be incorporated into lipid nanoemulsions (NE) is usually judged by their solubility in the internal phase of the formulation. This can lead to various problems, such as precipitation of the drug after processing the formulation or, depending on the preparation technique used, the use of a large amount of the drug. To this end, it is useful to consider other drug loading methods, especially in the early stages of formulation development. In this study, we aimed to find the best way to achieve the highest loading of GL-II -73 in NEs for future parenteral applications for in vivo animal studies. This ligand acts as a positive allosteric modulator at α-GABAA receptors with combined antidepressant and cognition enhancing effects. NEs were prepared using the high pressure homogenization technique, a standard technique for parenteral NE preparation. The oil phase (medium-chain triglycerides, soy lecithin, and butylated hydroxytoluene) and the aqueous phase (glycerol, polysorbate 80, and 0.01 M phosphate buffer, pH 8) were separately heated to 50 ˚C and mixed until all components were dissolved. The aqueous phase was added to the oil phase and processed first on a rotor-stator homogenizer at 11000 rpm for 1 minute and then on a high-pressure homogenizer at 800 bar for 10 cycles. This resulted in a droplet size of 117.1 ± 1.5 nm, a PDI of 0.060 ± 0.008, a zeta potential of - 43.3 ± 1.3 mV, a pH of 7.89 ± 0.02, and a conductivity of 1061.67 ± 5.51 S/cm, indicating initial suitability for parenteral use. Using the empirical method for drug loading, we were able to dissolve GL-II -73 in the oil phase and achieve a drug concentration of 1.5 mg/ml in NE. For our experimental setup, this required the use of 120 mg of GL-II -73 per experiment. For the passive drug loading procedure, we incubated 1 ml of the placebo NE in the eppendorf tube (in duplicate) with the excess drug (approximately 10 mg per tube) for 72 h. The drug was then added to the eppendorf tube. The excess of the undissolved drug was removed after centrifugation. The drug content in the supernatant was 3.10 ± 0.25 mg/ml, indicating above-average loading of the drug and possibly suggesting localization of the drug in the droplet-stabilising layer, but this needs to be further demonstrated. This approach could contribute to more rational formulation development in the selection of formulation factors.
Acknowledgments
This research was funded by the Science Fund of the Republic of Serbia, GRANT No 7749108, Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platform - NanoCellEmоCog.",
publisher = "International Association of Physical Chemists",
journal = "10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6",
title = "Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4999"
}

Đoković, J., Marković, B., Sharmin, D., Cook, J. M., Savić, M.,& Savić, S.. (2023). Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6
International Association of Physical Chemists..
https://hdl.handle.net/21.15107/rcub_farfar_4999

Đoković J, Marković B, Sharmin D, Cook JM, Savić M, Savić S. Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application. in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_4999 .

Đoković, Jelena, Marković, Bojan, Sharmin, Dishary, Cook, James M., Savić, Miroslav, Savić, Snežana, "Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application" in 10th IAPC Meeting Tenth World Conference on Physico-Chemical Methods in Drug Discovery & Sixth World Conference on ADMET and DMPK Belgrade, Serbia, September 4-6 (2023),
https://hdl.handle.net/21.15107/rcub_farfar_4999 .

TY  - JOUR
AU  - Mitrović, Jelena
AU  - Bjelošević Žiberna, Maja
AU  - Vukadinović, Aleksandar
AU  - Knutson, Daniel E.
AU  - Sharmin, Dishary
AU  - Kremenović, Aleksandar
AU  - Ahlin Grabnar, Pegi
AU  - Planinšek, Odon
AU  - Lunter, Dominique
AU  - Cook, James M
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4982
AB  - Recently, nanocrystal dispersions have been considered as a promising formulation strategy to improve the bioavailability of the deuterated pyrazoloquinolinone ligand DK-I-56-1 (7‑methoxy-2-(4‑methoxy-d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one). In the current study, the freeze-drying process (formulation and process parameters) was investigated to improve the storage stability of the previously developed formulation. Different combinations of lyoprotectant (sucrose or trehalose) and bulking agent (mannitol) were varied while formulations were freeze-dried under two conditions (primary drying at -10 or -45 °C). The obtained lyophilizates were characterized in terms of particle size, solid state properties and morphology, while the interactions within the samples were analyzed by Fourier transform infrared spectroscopy. In the preliminary study, three formulations were selected based on the high redispersibility index values (around 95%). The temperature of primary drying had no significant effect on particle size, but stability during storage was impaired for samples dried at -10 °C. Samples dried at lower temperature were more homogeneous and remained stable for three months. It was found that the optimal ratio of sucrose or trehalose to mannitol was 3:2 at a total concentration of 10% to achieve the best stability (particle size < 1.0 μm, polydispersity index < 0.250). The amorphous state of lyoprotectants probably provided a high degree of interaction with nanocrystals, while the crystalline mannitol provided an elegant cake structure. Sucrose was superior to trehalose in maintaining particle size during freeze-drying, while trehalose was more effective in keeping particle size within limits during storage. In conclusion, results demonstrated that the appropriate combination of sucrose/trehalose and mannitol together with the appropriate selection of lyophilization process parameters could yield nanocrystals with satisfactory stability.
PB  - Elsevier B.V.
T2  - European Journal of Pharmaceutical Sciences
T1  - Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study
VL  - 189
DO  - 10.1016/j.ejps.2023.106557
ER  - 

@article{
author = "Mitrović, Jelena and Bjelošević Žiberna, Maja and Vukadinović, Aleksandar and Knutson, Daniel E. and Sharmin, Dishary and Kremenović, Aleksandar and Ahlin Grabnar, Pegi and Planinšek, Odon and Lunter, Dominique and Cook, James M and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Recently, nanocrystal dispersions have been considered as a promising formulation strategy to improve the bioavailability of the deuterated pyrazoloquinolinone ligand DK-I-56-1 (7‑methoxy-2-(4‑methoxy-d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one). In the current study, the freeze-drying process (formulation and process parameters) was investigated to improve the storage stability of the previously developed formulation. Different combinations of lyoprotectant (sucrose or trehalose) and bulking agent (mannitol) were varied while formulations were freeze-dried under two conditions (primary drying at -10 or -45 °C). The obtained lyophilizates were characterized in terms of particle size, solid state properties and morphology, while the interactions within the samples were analyzed by Fourier transform infrared spectroscopy. In the preliminary study, three formulations were selected based on the high redispersibility index values (around 95%). The temperature of primary drying had no significant effect on particle size, but stability during storage was impaired for samples dried at -10 °C. Samples dried at lower temperature were more homogeneous and remained stable for three months. It was found that the optimal ratio of sucrose or trehalose to mannitol was 3:2 at a total concentration of 10% to achieve the best stability (particle size < 1.0 μm, polydispersity index < 0.250). The amorphous state of lyoprotectants probably provided a high degree of interaction with nanocrystals, while the crystalline mannitol provided an elegant cake structure. Sucrose was superior to trehalose in maintaining particle size during freeze-drying, while trehalose was more effective in keeping particle size within limits during storage. In conclusion, results demonstrated that the appropriate combination of sucrose/trehalose and mannitol together with the appropriate selection of lyophilization process parameters could yield nanocrystals with satisfactory stability.",
publisher = "Elsevier B.V.",
journal = "European Journal of Pharmaceutical Sciences",
title = "Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study",
volume = "189",
doi = "10.1016/j.ejps.2023.106557"
}

Mitrović, J., Bjelošević Žiberna, M., Vukadinović, A., Knutson, D. E., Sharmin, D., Kremenović, A., Ahlin Grabnar, P., Planinšek, O., Lunter, D., Cook, J. M., Savić, M.,& Savić, S.. (2023). Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study. in European Journal of Pharmaceutical Sciences
Elsevier B.V.., 189.
https://doi.org/10.1016/j.ejps.2023.106557

Mitrović J, Bjelošević Žiberna M, Vukadinović A, Knutson DE, Sharmin D, Kremenović A, Ahlin Grabnar P, Planinšek O, Lunter D, Cook JM, Savić M, Savić S. Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study. in European Journal of Pharmaceutical Sciences. 2023;189.
doi:10.1016/j.ejps.2023.106557 .

Mitrović, Jelena, Bjelošević Žiberna, Maja, Vukadinović, Aleksandar, Knutson, Daniel E., Sharmin, Dishary, Kremenović, Aleksandar, Ahlin Grabnar, Pegi, Planinšek, Odon, Lunter, Dominique, Cook, James M, Savić, Miroslav, Savić, Snežana, "Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study" in European Journal of Pharmaceutical Sciences, 189 (2023),
https://doi.org/10.1016/j.ejps.2023.106557 . .

TY  - JOUR
AU  - Stanković, Dragana
AU  - Radović, Magdalena
AU  - Stanković, Aljoša
AU  - Mirković, Marija
AU  - Vukadinović, Aleksandar
AU  - Mijović, Milica
AU  - Milanović, Zorana
AU  - Ognjanović, Miloš
AU  - Janković, Drina
AU  - Antić, Bratislav
AU  - Vranješ-Đurić, Sanja
AU  - Savić, Miroslav
AU  - Prijović, Željko
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4962
AB  - As an alternative to classical brachytherapy, intratumoral injection of radionuclide-labeled nanoparticles (nanobrachytherapy, NBT) has been investigated as a superior delivery method over an intravenous route for radionuclide therapy of solid tumors. We created superparamagnetic iron oxide nanoparticles (SPIONs) coated with meso-1,2-dimercaptosuccinic acid (DMSA) and radiola- beled with Lutetium-177 (177Lu), generating 177Lu-DMSA@SPIONs as a potential antitumor agent for nanobrachytherapy. Efficient radiolabeling of DMSA@SPIONS by 177Lu resulted in a stable bond with minimal leakage in vitro. After an intratumoral injection to mouse colorectal CT-26 or breast 4T1 subcutaneous tumors, the nanoparticles remained well localized at the injection site for weeks, with limited leakage. The dose of 3.70 MBq/100 μg/50 μL of 177Lu-DMSA@SPIONs applied intratumorally resulted in a high therapeutic efficacy, without signs of general toxicity. A decreased dose of 1.85 MBq/100 μg/50 μL still retained therapeutic efficacy, while an increased dose of 9.25 MBq/100 μg/50 μL did not significantly benefit the therapy. Histopathology analysis revealed that the 177Lu-DMSA@SPIONs act within a limited range around the injection site, which explains the good therapeutic efficacy achieved by a single administration of a relatively low dose without the need for increased or repeated dosing. Overall, 177Lu-DMSA@SPIONs are safe and potent agents suitable for intra-tumoral administration for localized tumor radionuclide therapy.
PB  - MDPI
T2  - Pharmaceutics
T1  - Synthesis, Characterization, and Therapeutic Efficacy of 177Lu-DMSA@SPIONs in Nanobrachytherapy of Solid Tumors
VL  - 15
IS  - 7
DO  - 10.3390/pharmaceutics15071943
ER  - 

@article{
author = "Stanković, Dragana and Radović, Magdalena and Stanković, Aljoša and Mirković, Marija and Vukadinović, Aleksandar and Mijović, Milica and Milanović, Zorana and Ognjanović, Miloš and Janković, Drina and Antić, Bratislav and Vranješ-Đurić, Sanja and Savić, Miroslav and Prijović, Željko",
year = "2023",
abstract = "As an alternative to classical brachytherapy, intratumoral injection of radionuclide-labeled nanoparticles (nanobrachytherapy, NBT) has been investigated as a superior delivery method over an intravenous route for radionuclide therapy of solid tumors. We created superparamagnetic iron oxide nanoparticles (SPIONs) coated with meso-1,2-dimercaptosuccinic acid (DMSA) and radiola- beled with Lutetium-177 (177Lu), generating 177Lu-DMSA@SPIONs as a potential antitumor agent for nanobrachytherapy. Efficient radiolabeling of DMSA@SPIONS by 177Lu resulted in a stable bond with minimal leakage in vitro. After an intratumoral injection to mouse colorectal CT-26 or breast 4T1 subcutaneous tumors, the nanoparticles remained well localized at the injection site for weeks, with limited leakage. The dose of 3.70 MBq/100 μg/50 μL of 177Lu-DMSA@SPIONs applied intratumorally resulted in a high therapeutic efficacy, without signs of general toxicity. A decreased dose of 1.85 MBq/100 μg/50 μL still retained therapeutic efficacy, while an increased dose of 9.25 MBq/100 μg/50 μL did not significantly benefit the therapy. Histopathology analysis revealed that the 177Lu-DMSA@SPIONs act within a limited range around the injection site, which explains the good therapeutic efficacy achieved by a single administration of a relatively low dose without the need for increased or repeated dosing. Overall, 177Lu-DMSA@SPIONs are safe and potent agents suitable for intra-tumoral administration for localized tumor radionuclide therapy.",
publisher = "MDPI",
journal = "Pharmaceutics",
title = "Synthesis, Characterization, and Therapeutic Efficacy of 177Lu-DMSA@SPIONs in Nanobrachytherapy of Solid Tumors",
volume = "15",
number = "7",
doi = "10.3390/pharmaceutics15071943"
}

Stanković, D., Radović, M., Stanković, A., Mirković, M., Vukadinović, A., Mijović, M., Milanović, Z., Ognjanović, M., Janković, D., Antić, B., Vranješ-Đurić, S., Savić, M.,& Prijović, Ž.. (2023). Synthesis, Characterization, and Therapeutic Efficacy of 177Lu-DMSA@SPIONs in Nanobrachytherapy of Solid Tumors. in Pharmaceutics
MDPI., 15(7).
https://doi.org/10.3390/pharmaceutics15071943

Stanković D, Radović M, Stanković A, Mirković M, Vukadinović A, Mijović M, Milanović Z, Ognjanović M, Janković D, Antić B, Vranješ-Đurić S, Savić M, Prijović Ž. Synthesis, Characterization, and Therapeutic Efficacy of 177Lu-DMSA@SPIONs in Nanobrachytherapy of Solid Tumors. in Pharmaceutics. 2023;15(7).
doi:10.3390/pharmaceutics15071943 .

Stanković, Dragana, Radović, Magdalena, Stanković, Aljoša, Mirković, Marija, Vukadinović, Aleksandar, Mijović, Milica, Milanović, Zorana, Ognjanović, Miloš, Janković, Drina, Antić, Bratislav, Vranješ-Đurić, Sanja, Savić, Miroslav, Prijović, Željko, "Synthesis, Characterization, and Therapeutic Efficacy of 177Lu-DMSA@SPIONs in Nanobrachytherapy of Solid Tumors" in Pharmaceutics, 15, no. 7 (2023),
https://doi.org/10.3390/pharmaceutics15071943 . .

TY  - JOUR
AU  - Mirković, Duško
AU  - Beletić, Anđelo
AU  - Savić, Miroslav
AU  - Milinković, Neda
AU  - Sarić-Matutinović, Marija
AU  - Jančić, Ivan
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4910
AB  - Occupational and environmental toxicology specialists find catecholamine fluctuations in brain tissue relevant for research of neurotoxicity, such as that induced by manganese or zinc, pesticides, industrial solvents, plastic, air pollution, or irradiation. Considering that catecholamine tissue concentrations are generally very low, their extraction requires a reliable and optimal method that will achieve maximum recovery and minimise other interferences. This study aimed to evaluate whether the aluminium (III) oxide (Al2O3, alumina) based cartridges designed for catecholamine isolation from plasma could be used for solid-phase extraction (SPE) of catecholamine from the brain tissue. To do that, we homogenised Wistar rat brain tissue with perchloric acid and compared three extraction techniques: SPE, the routine filtration through a 0.22 µm membrane filter, and their combination. In the extracts, we compared relative chromatographic catecholamine mobility measured with high performance liquid chromatography with electrochemical detection. Chromatographic patterns for norepinephrine and epinephrine were similar regardless of the extraction technique, which indicates that the alumina cartridge is good enough to isolate them from brain tissue. However, the dopamine pattern was unsatisfactory, and further experiments are needed to identify the issue and optimise the protocol. © 2023 Duško Mirković et al., published by Sciendo.  Promjene razine katekolamina (KAT) u moždanom tkivu značajne su za brojna područja profesionalne toksikologije odnosno ekotoksikologije u kojima se istražuje neurotoksičnost izazvana različitim agensima poput mangana ili cinka, pesticida, industrijskih otapala, plastike, aerozagađenja ili zračenja. Niske koncentracije KAT-a u tkivu zahtijevaju pouzdanu i učinkovitu tehniku ekstrakcije kojom se postiže maksimalni „prinos“ katekolamina i minimalni sadržaj interferirajućih spojeva. Cilj istraživanja bio je procijeniti mogu li se kolone na bazi aluminijeva (III) oksida (Al2O3), dizajnirane za izolaciju KAT-a iz plazme, koristiti za ekstrakciju čvrstom fazom (eng. solid-phase extraction – SPE) KAT-a iz moždanoga tkiva. Nakon homogenizacije tkiva Sprague Dawley štakora upotrebom perklorne kiseline, primijenjene su tri tehnike ekstrakcije: SPE, filtracija kroz 0,22 µm membranski filtar, koji je zapravo rutinska tehnika za izolaciju KAT-a iz mozga, i kombinacija tih dviju tehnika. U ekstraktima je relativna kromatografska pokretljivost KAT-a analizirana HPLC metodom s elektrokemijskom detekcijom. Ponašanje norepinefrina i epinefrina tijekom kromatografije bilo je slično, bez obzira na tehniku ekstrakcije, što upućuje na to da aluminijev oksid ima zadovoljavajuća svojstva izolirati ta dva KAT-a iz moždanoga tkiva. Međutim, uočeni su problemi s ekstrakcijom dopamina, koji zahtijevaju dodatne eksperimente kako bi se otkrio njihov uzrok i osmislio protokol optimizacije. Author keywords aluminijev oksid; aluminium oxide; brain; catecholamines; ekstrakcija čvrste faze; katekolamini; mozak; solid phase extraction; tissue; tkivo   © This record is sourced from MEDLINE/PubMed, a database of the U.S. National Library of Medicine      1of1   Top of page Cited by 0 documents Inform me when this document is cited in Scopus: Related documents Find more related documents in Scopus based on: Authors Keywords
AB  - Promjene razine katekolamina (KAT) u moždanom tkivu značajne su za brojna područja profesionalne toksikologije odnosno ekotoksikologije u kojima se istražuje neurotoksičnost izazvana različitim agensima poput mangana ili cinka, pesticida, industrijskih otapala, plastike, aerozagađenja ili zračenja. Niske koncentracije KAT-a u tkivu zahtijevaju pouzdanu i učinkovitu tehniku ekstrakcije kojom se postiže maksimalni „prinos“ katekolamina i minimalni sadržaj interferirajućih spojeva. Cilj istraživanja bio je procijeniti mogu li se kolone na bazi aluminijeva (III) oksida (Al2 O 3), dizajnirane za izolaciju KAT-a iz plazme, koristiti za ekstrakciju čvrstom fazom (eng. solid-phase extraction – SPE) KAT-a iz moždanoga tkiva. Nakon homogenizacije tkiva Sprague Dawley štakora upotrebom perklorne kiseline, primijenjene su tri tehnike ekstrakcije: SPE, filtracija kroz 0,22 μm membranski filtar, koji je zapravo rutinska tehnika za izolaciju KAT-a iz mozga, i kombinacija tih dviju tehnika. U ekstraktima je relativna kromatografska pokretljivost KAT-a analizirana HPLC metodom s elektrokemijskom detekcijom. Ponašanje norepinefrina i epinefrina tijekom kromatografije bilo je slično, bez obzira na tehniku ekstrakcije, što upućuje na to da aluminijev oksid ima zadovoljavajuća svojstva izolirati ta dva KAT-a iz moždanoga tkiva. Međutim, uočeni su problemi s ekstrakcijom dopamina, koji zahtijevaju dodatne eksperimente kako bi se otkrio njihov uzrok i osmislio protokol optimizacije.
PB  - De Gruyter Poland
T2  - Arhiv za higijenu rada i toksikologiju
T2  - aluminijev oksid
T2  - ekstrakcija čvrste faze
T2  - katekolamini
T2  - mozak
T2  - tkivo
T1  - Is alumina suitable for solid phase extraction of catecholamines from brain tissue?
T1  - Je li aluminijev oksid pogodan za postupak ekstrakcije čvrste faze katekolamina iz moždanoga tkiva?
VL  - 74
IS  - 2
SP  - 120
EP  - 126
DO  - 10.2478/aiht-2023-74-3706
ER  - 

@article{
author = "Mirković, Duško and Beletić, Anđelo and Savić, Miroslav and Milinković, Neda and Sarić-Matutinović, Marija and Jančić, Ivan",
year = "2023",
abstract = "Occupational and environmental toxicology specialists find catecholamine fluctuations in brain tissue relevant for research of neurotoxicity, such as that induced by manganese or zinc, pesticides, industrial solvents, plastic, air pollution, or irradiation. Considering that catecholamine tissue concentrations are generally very low, their extraction requires a reliable and optimal method that will achieve maximum recovery and minimise other interferences. This study aimed to evaluate whether the aluminium (III) oxide (Al2O3, alumina) based cartridges designed for catecholamine isolation from plasma could be used for solid-phase extraction (SPE) of catecholamine from the brain tissue. To do that, we homogenised Wistar rat brain tissue with perchloric acid and compared three extraction techniques: SPE, the routine filtration through a 0.22 µm membrane filter, and their combination. In the extracts, we compared relative chromatographic catecholamine mobility measured with high performance liquid chromatography with electrochemical detection. Chromatographic patterns for norepinephrine and epinephrine were similar regardless of the extraction technique, which indicates that the alumina cartridge is good enough to isolate them from brain tissue. However, the dopamine pattern was unsatisfactory, and further experiments are needed to identify the issue and optimise the protocol. © 2023 Duško Mirković et al., published by Sciendo.  Promjene razine katekolamina (KAT) u moždanom tkivu značajne su za brojna područja profesionalne toksikologije odnosno ekotoksikologije u kojima se istražuje neurotoksičnost izazvana različitim agensima poput mangana ili cinka, pesticida, industrijskih otapala, plastike, aerozagađenja ili zračenja. Niske koncentracije KAT-a u tkivu zahtijevaju pouzdanu i učinkovitu tehniku ekstrakcije kojom se postiže maksimalni „prinos“ katekolamina i minimalni sadržaj interferirajućih spojeva. Cilj istraživanja bio je procijeniti mogu li se kolone na bazi aluminijeva (III) oksida (Al2O3), dizajnirane za izolaciju KAT-a iz plazme, koristiti za ekstrakciju čvrstom fazom (eng. solid-phase extraction – SPE) KAT-a iz moždanoga tkiva. Nakon homogenizacije tkiva Sprague Dawley štakora upotrebom perklorne kiseline, primijenjene su tri tehnike ekstrakcije: SPE, filtracija kroz 0,22 µm membranski filtar, koji je zapravo rutinska tehnika za izolaciju KAT-a iz mozga, i kombinacija tih dviju tehnika. U ekstraktima je relativna kromatografska pokretljivost KAT-a analizirana HPLC metodom s elektrokemijskom detekcijom. Ponašanje norepinefrina i epinefrina tijekom kromatografije bilo je slično, bez obzira na tehniku ekstrakcije, što upućuje na to da aluminijev oksid ima zadovoljavajuća svojstva izolirati ta dva KAT-a iz moždanoga tkiva. Međutim, uočeni su problemi s ekstrakcijom dopamina, koji zahtijevaju dodatne eksperimente kako bi se otkrio njihov uzrok i osmislio protokol optimizacije. Author keywords aluminijev oksid; aluminium oxide; brain; catecholamines; ekstrakcija čvrste faze; katekolamini; mozak; solid phase extraction; tissue; tkivo   © This record is sourced from MEDLINE/PubMed, a database of the U.S. National Library of Medicine      1of1   Top of page Cited by 0 documents Inform me when this document is cited in Scopus: Related documents Find more related documents in Scopus based on: Authors Keywords, Promjene razine katekolamina (KAT) u moždanom tkivu značajne su za brojna područja profesionalne toksikologije odnosno ekotoksikologije u kojima se istražuje neurotoksičnost izazvana različitim agensima poput mangana ili cinka, pesticida, industrijskih otapala, plastike, aerozagađenja ili zračenja. Niske koncentracije KAT-a u tkivu zahtijevaju pouzdanu i učinkovitu tehniku ekstrakcije kojom se postiže maksimalni „prinos“ katekolamina i minimalni sadržaj interferirajućih spojeva. Cilj istraživanja bio je procijeniti mogu li se kolone na bazi aluminijeva (III) oksida (Al2 O 3), dizajnirane za izolaciju KAT-a iz plazme, koristiti za ekstrakciju čvrstom fazom (eng. solid-phase extraction – SPE) KAT-a iz moždanoga tkiva. Nakon homogenizacije tkiva Sprague Dawley štakora upotrebom perklorne kiseline, primijenjene su tri tehnike ekstrakcije: SPE, filtracija kroz 0,22 μm membranski filtar, koji je zapravo rutinska tehnika za izolaciju KAT-a iz mozga, i kombinacija tih dviju tehnika. U ekstraktima je relativna kromatografska pokretljivost KAT-a analizirana HPLC metodom s elektrokemijskom detekcijom. Ponašanje norepinefrina i epinefrina tijekom kromatografije bilo je slično, bez obzira na tehniku ekstrakcije, što upućuje na to da aluminijev oksid ima zadovoljavajuća svojstva izolirati ta dva KAT-a iz moždanoga tkiva. Međutim, uočeni su problemi s ekstrakcijom dopamina, koji zahtijevaju dodatne eksperimente kako bi se otkrio njihov uzrok i osmislio protokol optimizacije.",
publisher = "De Gruyter Poland",
journal = "Arhiv za higijenu rada i toksikologiju, aluminijev oksid, ekstrakcija čvrste faze, katekolamini, mozak, tkivo",
title = "Is alumina suitable for solid phase extraction of catecholamines from brain tissue?, Je li aluminijev oksid pogodan za postupak ekstrakcije čvrste faze katekolamina iz moždanoga tkiva?",
volume = "74",
number = "2",
pages = "120-126",
doi = "10.2478/aiht-2023-74-3706"
}

Mirković, D., Beletić, A., Savić, M., Milinković, N., Sarić-Matutinović, M.,& Jančić, I.. (2023). Is alumina suitable for solid phase extraction of catecholamines from brain tissue?. in Arhiv za higijenu rada i toksikologiju
De Gruyter Poland., 74(2), 120-126.
https://doi.org/10.2478/aiht-2023-74-3706

Mirković D, Beletić A, Savić M, Milinković N, Sarić-Matutinović M, Jančić I. Is alumina suitable for solid phase extraction of catecholamines from brain tissue?. in Arhiv za higijenu rada i toksikologiju. 2023;74(2):120-126.
doi:10.2478/aiht-2023-74-3706 .

Mirković, Duško, Beletić, Anđelo, Savić, Miroslav, Milinković, Neda, Sarić-Matutinović, Marija, Jančić, Ivan, "Is alumina suitable for solid phase extraction of catecholamines from brain tissue?" in Arhiv za higijenu rada i toksikologiju, 74, no. 2 (2023):120-126,
https://doi.org/10.2478/aiht-2023-74-3706 . .

TY  - CONF
AU  - Ilić, Tanja
AU  - Stanković, Tijana
AU  - Mitrović, Jelena
AU  - Pantelić, Ivana
AU  - Dobričić, Vladimir
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4583
AB  - INTRODUCTION
Recently, the modulation of sigma-2 receptors localized in the brain is proposed to be linked with regulation of mood, anxiety, and cognition [1]. Hence, we hypothesized that novel patent-protected ligand of the pyrazoloquinolinone chemotype (CW-02-79) with a substantial binding affinity for sigma-2 receptors may have a distinct pharmacological profile useful for the treatment of mood, anxiety, and/or cognitive symptoms that usually accompany numerous psychiatric and neurodegenerative disorders. Having in mind that the neuroimmune mechanisms play an important role in pathogenesis of various emotional and cognitive impairments, we aim to test whether modulation of sigma-2 receptors with CW-02-79 results in substantial improvements in neuroimmune and/or behavioral outputs in in vitro cell platforms consisting of human induced pluripotent stem cells and in vivo animal models made to mimic a compromised neuroimmune status. However, very low water solubility of CW-02-79 hinders its administration and reliable efficacy and safety in vitro/in vivo evaluation. In order to avoid usage of non-physiological solvents/vehicles such as dimethyl sulfoxide and consequently, vehicle-related safety issues, nanoemulsions based on biocompatible excipients could be a promising tool for effective preclinical testing of the selected drug candidate. Therefore, firstly, this study aimed to develop biocompatible nanoemulsions (NEs), as carrier for CW-02-79, tailored for the described preclinical studies, using high pressure homogenization (HPH) method. As a first step, preformulation studies were performed to obtain insight into the key properties of CW-02-79 required for further stages of formulation development. Afterward, during NE preparation, the influence of formulation and process parameters on particle size was investigated to obtain NEs with small and uniform particle size suitable for parenteral administration.
EXPERIMENTAL METHODS
Materials
For the preparation of NEs the following ingredients were used: CW-02-79 (synthesized at the Department of Chemistry and Biochemistry, University of Wisconsin—Milwaukee, WI, USA), medium-chain triglycerides (MCT) (Fagron GmbH & KG, Germany), castor oil, polysorbate 80, butylhydroxytoluene, glycerol (Sigma-Aldrich GmbH, Germany), soybean lecithin (Lipoid S75; Lipoid GmbH, Germany) and ultrapure water.
Preformulation Studies
The solubility of CW-02-79 in different oils and oil mixtures, distilled water, 0.1 M hydrochloride acid, phosphate buffer (pH 7.4), commonly used organic solvents (isopropanol, methanol and dimethyl sulfoxide) at 25 °C was investigated by the shake flask method. CW-02-79 concentration in the obtained supernatants was measured by LC-MS/MS. To gain certain insight into the physical state of CW-02-79, polarization microscopy and differential scanning calorimetry (DSC 1, Mettler–Toledo AG, Switzerland) were used.
Preparation and Characterization of NEs
Blank and CW-02-79-loaded NEs were prepared by varying the content of the oil phase (20%/30%, w/w) and process parameters (number of homogenization cycles), using hot HPH (EmulsiFlex-C3, Avestin Inc., Canada) at 800 bar and 50°C. The oil to surfactant ratio was kept constant (5:1, w/w) in all tested formulations. Droplet size (Z-ave), polydispersity index (PDI) and zeta potential (ZP) of corresponding NEs, after proper dilution, were determined using Zetasizer Nano ZS90 (Malvern Instruments Ltd., UK). Conductivity and pH value were measured by the conductometer (CDM230 Radiometer, Denmark) and pH meter (HI 9321, Hanna Instruments Inc, USA), respectively.
RESULTS AND DISCUSSION
Substance CW-02-79 appeared as a yellow powder, with broad particle size distribution. Results of the solubility study showed that, among the tested oils, the highest solubility of CW-02-79 was achieved in MCT-castor oil mixture (1:1, w/w) which was chosen as the oil phase for NE development. Elevated temperature (50°C) and presence of soybean lecithin as a solubilizer contributed to the loading of the target 2 mg/ml concentration, without precipitation during the storage.
After the oil phase selection, blank and CW-02-79-loaded NEs were prepared by varying the content of oil phase, 20% and 30%, w/w (increasing the oil content would reduce the volume to be injected). Polysorbate 80 was added as an additional stabilizer and functional excipient due to its tendency to enhance brain uptake of drugs by acting as P-glycoprotein inhibitor, stealth agent or promoter of receptor-mediated endocytosis [2]. Simultaneously, the impact of the number of homogenization cycles on critical quality attributes of NEs (Z-ave and PDI) was tested.
The observed increase in droplet size distribution (Figure 1) with increasing the number of homogenization cycles (> 8 cycles) could be attributed to over-processing (probably caused by increased droplet collision and re-coalescence rates or by insufficient emulsifier concentration in relation to the increasing interfacial area). Interestingly, although larger oil volume fractions generally lead to increased droplet collisions and hence larger droplet size [3], no statistically significant difference regarding droplet size was observed between formulations prepared with 20 and 30% of the oil phase (at 7 HPH cycles, 800 bar, 50°C). Likewise, a relatively narrow particle size distribution (PDI < 0.15) was observed, suggesting that the developed NEs were suitable for parenteral application. Moreover, satisfactory values were observed for all other tested physicochemical parameters (Table 2). Absolute ZP values were above 30 mV, indicating good stability of the system. Furthermore, the incorporation of CW-02-79 did not exert any influence on NE physicochemical properties, irrespective of the oil content.
In conclusion, although the formulation prepared with 30% of the oil phase had satisfying physicochemical properties, its relatively high viscosity can restrict syringeability and injectability. On the other hand, owing to satisfying solubilization capacity for CW-02-79 as well as small and uniform droplet size and low viscosity, NE prepared with 20% of the oil phase represents a promising carrier worth exploring further to support the preclinical progress of CW-02-79.
C3  - 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France
T1  - Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4583
ER  - 

@conference{
author = "Ilić, Tanja and Stanković, Tijana and Mitrović, Jelena and Pantelić, Ivana and Dobričić, Vladimir and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "INTRODUCTION
Recently, the modulation of sigma-2 receptors localized in the brain is proposed to be linked with regulation of mood, anxiety, and cognition [1]. Hence, we hypothesized that novel patent-protected ligand of the pyrazoloquinolinone chemotype (CW-02-79) with a substantial binding affinity for sigma-2 receptors may have a distinct pharmacological profile useful for the treatment of mood, anxiety, and/or cognitive symptoms that usually accompany numerous psychiatric and neurodegenerative disorders. Having in mind that the neuroimmune mechanisms play an important role in pathogenesis of various emotional and cognitive impairments, we aim to test whether modulation of sigma-2 receptors with CW-02-79 results in substantial improvements in neuroimmune and/or behavioral outputs in in vitro cell platforms consisting of human induced pluripotent stem cells and in vivo animal models made to mimic a compromised neuroimmune status. However, very low water solubility of CW-02-79 hinders its administration and reliable efficacy and safety in vitro/in vivo evaluation. In order to avoid usage of non-physiological solvents/vehicles such as dimethyl sulfoxide and consequently, vehicle-related safety issues, nanoemulsions based on biocompatible excipients could be a promising tool for effective preclinical testing of the selected drug candidate. Therefore, firstly, this study aimed to develop biocompatible nanoemulsions (NEs), as carrier for CW-02-79, tailored for the described preclinical studies, using high pressure homogenization (HPH) method. As a first step, preformulation studies were performed to obtain insight into the key properties of CW-02-79 required for further stages of formulation development. Afterward, during NE preparation, the influence of formulation and process parameters on particle size was investigated to obtain NEs with small and uniform particle size suitable for parenteral administration.
EXPERIMENTAL METHODS
Materials
For the preparation of NEs the following ingredients were used: CW-02-79 (synthesized at the Department of Chemistry and Biochemistry, University of Wisconsin—Milwaukee, WI, USA), medium-chain triglycerides (MCT) (Fagron GmbH & KG, Germany), castor oil, polysorbate 80, butylhydroxytoluene, glycerol (Sigma-Aldrich GmbH, Germany), soybean lecithin (Lipoid S75; Lipoid GmbH, Germany) and ultrapure water.
Preformulation Studies
The solubility of CW-02-79 in different oils and oil mixtures, distilled water, 0.1 M hydrochloride acid, phosphate buffer (pH 7.4), commonly used organic solvents (isopropanol, methanol and dimethyl sulfoxide) at 25 °C was investigated by the shake flask method. CW-02-79 concentration in the obtained supernatants was measured by LC-MS/MS. To gain certain insight into the physical state of CW-02-79, polarization microscopy and differential scanning calorimetry (DSC 1, Mettler–Toledo AG, Switzerland) were used.
Preparation and Characterization of NEs
Blank and CW-02-79-loaded NEs were prepared by varying the content of the oil phase (20%/30%, w/w) and process parameters (number of homogenization cycles), using hot HPH (EmulsiFlex-C3, Avestin Inc., Canada) at 800 bar and 50°C. The oil to surfactant ratio was kept constant (5:1, w/w) in all tested formulations. Droplet size (Z-ave), polydispersity index (PDI) and zeta potential (ZP) of corresponding NEs, after proper dilution, were determined using Zetasizer Nano ZS90 (Malvern Instruments Ltd., UK). Conductivity and pH value were measured by the conductometer (CDM230 Radiometer, Denmark) and pH meter (HI 9321, Hanna Instruments Inc, USA), respectively.
RESULTS AND DISCUSSION
Substance CW-02-79 appeared as a yellow powder, with broad particle size distribution. Results of the solubility study showed that, among the tested oils, the highest solubility of CW-02-79 was achieved in MCT-castor oil mixture (1:1, w/w) which was chosen as the oil phase for NE development. Elevated temperature (50°C) and presence of soybean lecithin as a solubilizer contributed to the loading of the target 2 mg/ml concentration, without precipitation during the storage.
After the oil phase selection, blank and CW-02-79-loaded NEs were prepared by varying the content of oil phase, 20% and 30%, w/w (increasing the oil content would reduce the volume to be injected). Polysorbate 80 was added as an additional stabilizer and functional excipient due to its tendency to enhance brain uptake of drugs by acting as P-glycoprotein inhibitor, stealth agent or promoter of receptor-mediated endocytosis [2]. Simultaneously, the impact of the number of homogenization cycles on critical quality attributes of NEs (Z-ave and PDI) was tested.
The observed increase in droplet size distribution (Figure 1) with increasing the number of homogenization cycles (> 8 cycles) could be attributed to over-processing (probably caused by increased droplet collision and re-coalescence rates or by insufficient emulsifier concentration in relation to the increasing interfacial area). Interestingly, although larger oil volume fractions generally lead to increased droplet collisions and hence larger droplet size [3], no statistically significant difference regarding droplet size was observed between formulations prepared with 20 and 30% of the oil phase (at 7 HPH cycles, 800 bar, 50°C). Likewise, a relatively narrow particle size distribution (PDI < 0.15) was observed, suggesting that the developed NEs were suitable for parenteral application. Moreover, satisfactory values were observed for all other tested physicochemical parameters (Table 2). Absolute ZP values were above 30 mV, indicating good stability of the system. Furthermore, the incorporation of CW-02-79 did not exert any influence on NE physicochemical properties, irrespective of the oil content.
In conclusion, although the formulation prepared with 30% of the oil phase had satisfying physicochemical properties, its relatively high viscosity can restrict syringeability and injectability. On the other hand, owing to satisfying solubilization capacity for CW-02-79 as well as small and uniform droplet size and low viscosity, NE prepared with 20% of the oil phase represents a promising carrier worth exploring further to support the preclinical progress of CW-02-79.",
journal = "4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France",
title = "Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4583"
}

Ilić, T., Stanković, T., Mitrović, J., Pantelić, I., Dobričić, V., Cook, J. M., Savić, M.,& Savić, S.. (2023). Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France.
https://hdl.handle.net/21.15107/rcub_farfar_4583

Ilić T, Stanković T, Mitrović J, Pantelić I, Dobričić V, Cook JM, Savić M, Savić S. Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_4583 .

Ilić, Tanja, Stanković, Tijana, Mitrović, Jelena, Pantelić, Ivana, Dobričić, Vladimir, Cook, James M., Savić, Miroslav, Savić, Snežana, "Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies" in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_4583 .

TY  - JOUR
AU  - Ping, Xingjie
AU  - Meyer, Michelle J.
AU  - Zahn, Nicolas M.
AU  - Golani, Lalit K.
AU  - Sharmin, Dishary
AU  - Pandey, Kamal P.
AU  - Revanian, Sepideh
AU  - Mondal, Prithu
AU  - Jin, Xiaoming
AU  - Arnold, Leggy A.
AU  - Cerne, Rok
AU  - Cook, James M.
AU  - Divović, Branka
AU  - Savić, Miroslav
AU  - Lippa, Arnold
AU  - Smith, Jodi L.
AU  - Witkin, Jeffrey M.
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4427
AB  - A series of imidazodiazepines has been developed that possess reduced sedative liabilities but retain efficacy in anticonvulsant screening models. The latest of these compounds, (5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazole[1,5-α][1,4]diazepin-3-yl) oxazole known as KRM-II-81) is currently awaiting advancement into the clinic. A deuterated structural analog (D5-KRM-II-81) was made as a potential backup compound and studied here in comparison to KRM-II-81. In the present study, both compounds significantly prevented seizures in mice induced by 6 Hz (44 mA) electrical stimulation without significantly altering motoric function on a rotarod after intraperitoneal administration. Both compounds also significantly prevented clonic seizures, tonic seizures, and lethality induced by pentylenetetrazol in mice when given orally. D5-KRM-II-81 had a slightly longer duration of action against clonic and tonic seizures than KRM-II-81. Oral administration of 100 mg/kg of either KRM-II-81 or D5-KRM-II-81 was significantly less disruptive of sensorimotor function in mice than diazepam (5 mg/kg, p.o.). The present report documents that D5-KRM-II-81 represents another in this series of imidazodiazepines with anticonvulsant activity at doses that do not impair sensorimotor function.
PB  - John Wiley and Sons Inc
T2  - Drug Development Research
T1  - Comparative anticonvulsant activity of the GABAkine KRM-II-81 and a deuterated analog
VL  - 84
IS  - 3
DO  - 10.1002/ddr.22042
ER  - 

@article{
author = "Ping, Xingjie and Meyer, Michelle J. and Zahn, Nicolas M. and Golani, Lalit K. and Sharmin, Dishary and Pandey, Kamal P. and Revanian, Sepideh and Mondal, Prithu and Jin, Xiaoming and Arnold, Leggy A. and Cerne, Rok and Cook, James M. and Divović, Branka and Savić, Miroslav and Lippa, Arnold and Smith, Jodi L. and Witkin, Jeffrey M.",
year = "2023",
abstract = "A series of imidazodiazepines has been developed that possess reduced sedative liabilities but retain efficacy in anticonvulsant screening models. The latest of these compounds, (5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazole[1,5-α][1,4]diazepin-3-yl) oxazole known as KRM-II-81) is currently awaiting advancement into the clinic. A deuterated structural analog (D5-KRM-II-81) was made as a potential backup compound and studied here in comparison to KRM-II-81. In the present study, both compounds significantly prevented seizures in mice induced by 6 Hz (44 mA) electrical stimulation without significantly altering motoric function on a rotarod after intraperitoneal administration. Both compounds also significantly prevented clonic seizures, tonic seizures, and lethality induced by pentylenetetrazol in mice when given orally. D5-KRM-II-81 had a slightly longer duration of action against clonic and tonic seizures than KRM-II-81. Oral administration of 100 mg/kg of either KRM-II-81 or D5-KRM-II-81 was significantly less disruptive of sensorimotor function in mice than diazepam (5 mg/kg, p.o.). The present report documents that D5-KRM-II-81 represents another in this series of imidazodiazepines with anticonvulsant activity at doses that do not impair sensorimotor function.",
publisher = "John Wiley and Sons Inc",
journal = "Drug Development Research",
title = "Comparative anticonvulsant activity of the GABAkine KRM-II-81 and a deuterated analog",
volume = "84",
number = "3",
doi = "10.1002/ddr.22042"
}

Ping, X., Meyer, M. J., Zahn, N. M., Golani, L. K., Sharmin, D., Pandey, K. P., Revanian, S., Mondal, P., Jin, X., Arnold, L. A., Cerne, R., Cook, J. M., Divović, B., Savić, M., Lippa, A., Smith, J. L.,& Witkin, J. M.. (2023). Comparative anticonvulsant activity of the GABAkine KRM-II-81 and a deuterated analog. in Drug Development Research
John Wiley and Sons Inc., 84(3).
https://doi.org/10.1002/ddr.22042

Ping X, Meyer MJ, Zahn NM, Golani LK, Sharmin D, Pandey KP, Revanian S, Mondal P, Jin X, Arnold LA, Cerne R, Cook JM, Divović B, Savić M, Lippa A, Smith JL, Witkin JM. Comparative anticonvulsant activity of the GABAkine KRM-II-81 and a deuterated analog. in Drug Development Research. 2023;84(3).
doi:10.1002/ddr.22042 .

Ping, Xingjie, Meyer, Michelle J., Zahn, Nicolas M., Golani, Lalit K., Sharmin, Dishary, Pandey, Kamal P., Revanian, Sepideh, Mondal, Prithu, Jin, Xiaoming, Arnold, Leggy A., Cerne, Rok, Cook, James M., Divović, Branka, Savić, Miroslav, Lippa, Arnold, Smith, Jodi L., Witkin, Jeffrey M., "Comparative anticonvulsant activity of the GABAkine KRM-II-81 and a deuterated analog" in Drug Development Research, 84, no. 3 (2023),
https://doi.org/10.1002/ddr.22042 . .

TY  - GEN
AU  - Savić, Snežana
AU  - Pantelić, Ivana
AU  - Jančić, Ivan
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4298
AB  - Mood, anxiety and cognitive symptoms in psychiatry and neurology represent a significant worldwide burden. Due to difficulties in disease modeling and drug delivering to the site of action, as well as gaps in in vitro/in vivo extrapolation, the efforts to elucidate the roles of stress and neuroimmune pathways in both, etiology and therapy of these symptoms are challenging, but may nevertheless result in novel mechanisms of action. Recent preclinical studies provided novel leads/drug candidates with promising mood, anxiety and cognitive effects, the intellectual property rights of which are co-owned by the project beneficiary. We aim to: (1) incorporate the selective ligands of GABAA and/or sigma-2 receptors, with code names GL-II-73, DK-I-56, MM-I-03 and CW-02-79, together with two reference sigma-2 receptor ligands (siramesine and RHM-1), into the optimized nanoparticles and target their delivery to the human induced pluripotent stem cell (hiPSC)- based tri-culture cell neuroinflammation model, or rat brain; (2) quantify the immunological/morphological/neurochemical markers in immunologically challenged hiPSC-derived neurons, astrocytes and glia cells, and (3) assess their effects on behavior and biological markers in immunologically challenged animals of both sexes subjected to chronic mild unpredictable stress. We assume that the targeted nanodelivery of selected compounds to the brain will improve their pharmacokinetic profile, fortify their beneficial effect on mood, anxiety and cognition, and help delineate the contributing neuroimmune effects presumably arising mainly from microglia. The familiarization with neuroimmune aspects and pharmacokinetic optimization will support the preclinical progress of these compounds and might provide a rationale for designing clinical trials.
T2  - 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija
T1  - Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4298
ER  - 

@misc{
author = "Savić, Snežana and Pantelić, Ivana and Jančić, Ivan and Savić, Miroslav",
year = "2022",
abstract = "Mood, anxiety and cognitive symptoms in psychiatry and neurology represent a significant worldwide burden. Due to difficulties in disease modeling and drug delivering to the site of action, as well as gaps in in vitro/in vivo extrapolation, the efforts to elucidate the roles of stress and neuroimmune pathways in both, etiology and therapy of these symptoms are challenging, but may nevertheless result in novel mechanisms of action. Recent preclinical studies provided novel leads/drug candidates with promising mood, anxiety and cognitive effects, the intellectual property rights of which are co-owned by the project beneficiary. We aim to: (1) incorporate the selective ligands of GABAA and/or sigma-2 receptors, with code names GL-II-73, DK-I-56, MM-I-03 and CW-02-79, together with two reference sigma-2 receptor ligands (siramesine and RHM-1), into the optimized nanoparticles and target their delivery to the human induced pluripotent stem cell (hiPSC)- based tri-culture cell neuroinflammation model, or rat brain; (2) quantify the immunological/morphological/neurochemical markers in immunologically challenged hiPSC-derived neurons, astrocytes and glia cells, and (3) assess their effects on behavior and biological markers in immunologically challenged animals of both sexes subjected to chronic mild unpredictable stress. We assume that the targeted nanodelivery of selected compounds to the brain will improve their pharmacokinetic profile, fortify their beneficial effect on mood, anxiety and cognition, and help delineate the contributing neuroimmune effects presumably arising mainly from microglia. The familiarization with neuroimmune aspects and pharmacokinetic optimization will support the preclinical progress of these compounds and might provide a rationale for designing clinical trials.",
journal = "8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija",
title = "Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4298"
}

Savić, S., Pantelić, I., Jančić, I.,& Savić, M.. (2022). Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija.
https://hdl.handle.net/21.15107/rcub_farfar_4298

Savić S, Pantelić I, Jančić I, Savić M. Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija. 2022;.
https://hdl.handle.net/21.15107/rcub_farfar_4298 .

Savić, Snežana, Pantelić, Ivana, Jančić, Ivan, Savić, Miroslav, "Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms" in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija (2022),
https://hdl.handle.net/21.15107/rcub_farfar_4298 .

TY  - CONF
AU  - Mitrović, Jelena
AU  - Knutson, Daniel
AU  - Nikolić, Ines
AU  - Cook, James
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4285
AB  - After parenteral administration, nanoparticles interact with different proteins,
forming a shell called corona, which further influence nanoparticles’ biodistribution. Protein
adsorption is affected by particle size and shape, but also by molecular interactions of
chemical groups from the particle surface and amino-acid residues of the proteins. In human
plasma, albumin is the most abundant protein so it is frequently used for the investigation of
protein-nanoparticle interactions (1). In this study we investigated the attachment of bovine
serum albumin (BSA) to recently developed nanocrystals (2) of DK-I-56-1 (7-methoxy-2-
(4-methoxy-d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one), stabilized by
polysorbate 80 (NS2) or the combination of polysorbate 80 and poloxamer 407 (NS4).
Nanocrystal dispersion was incubated in medium containing 0.1% or 1% BSA in phosphate
buffer saline (pH 7,4) at 37 °C for 1 h. Particle size analysis was conducted by dynamic light
scattering in 10 min interval, at 37 °C on Zetasizer ZS90 (Malvern Instruments Ltd.,
Worcestershire, UK). It was shown that albumin adsorption was influenced by the
nanocrystal formulation and albumin concentration, but not incubation time. In a medium
with 0.1% BSA, no particle size difference was noticed in either formulation. However, in
case of NS2, after the addition of 1% albumin, particle size and particle size distribution
increased, which indicated albumin binding. On the other hand, in formulation NS4, with
higher albumin concentration two peaks were visible, one from the free albumin, and one
from nanocrystal particles. Therefore, it could be concluded that the affinity of albumin was
influenced mainly by the interaction with the nanocrystal stabilizers.
C3  - 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija
T1  - Investigation of albumin adsorption on DK-I-56-1 nanocrystals by dynamic light scattering
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4285
ER  - 

@conference{
author = "Mitrović, Jelena and Knutson, Daniel and Nikolić, Ines and Cook, James and Savić, Miroslav and Savić, Snežana",
year = "2022",
abstract = "After parenteral administration, nanoparticles interact with different proteins,
forming a shell called corona, which further influence nanoparticles’ biodistribution. Protein
adsorption is affected by particle size and shape, but also by molecular interactions of
chemical groups from the particle surface and amino-acid residues of the proteins. In human
plasma, albumin is the most abundant protein so it is frequently used for the investigation of
protein-nanoparticle interactions (1). In this study we investigated the attachment of bovine
serum albumin (BSA) to recently developed nanocrystals (2) of DK-I-56-1 (7-methoxy-2-
(4-methoxy-d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one), stabilized by
polysorbate 80 (NS2) or the combination of polysorbate 80 and poloxamer 407 (NS4).
Nanocrystal dispersion was incubated in medium containing 0.1% or 1% BSA in phosphate
buffer saline (pH 7,4) at 37 °C for 1 h. Particle size analysis was conducted by dynamic light
scattering in 10 min interval, at 37 °C on Zetasizer ZS90 (Malvern Instruments Ltd.,
Worcestershire, UK). It was shown that albumin adsorption was influenced by the
nanocrystal formulation and albumin concentration, but not incubation time. In a medium
with 0.1% BSA, no particle size difference was noticed in either formulation. However, in
case of NS2, after the addition of 1% albumin, particle size and particle size distribution
increased, which indicated albumin binding. On the other hand, in formulation NS4, with
higher albumin concentration two peaks were visible, one from the free albumin, and one
from nanocrystal particles. Therefore, it could be concluded that the affinity of albumin was
influenced mainly by the interaction with the nanocrystal stabilizers.",
journal = "8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija",
title = "Investigation of albumin adsorption on DK-I-56-1 nanocrystals by dynamic light scattering",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4285"
}

Mitrović, J., Knutson, D., Nikolić, I., Cook, J., Savić, M.,& Savić, S.. (2022). Investigation of albumin adsorption on DK-I-56-1 nanocrystals by dynamic light scattering. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija.
https://hdl.handle.net/21.15107/rcub_farfar_4285

Mitrović J, Knutson D, Nikolić I, Cook J, Savić M, Savić S. Investigation of albumin adsorption on DK-I-56-1 nanocrystals by dynamic light scattering. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija. 2022;.
https://hdl.handle.net/21.15107/rcub_farfar_4285 .

Mitrović, Jelena, Knutson, Daniel, Nikolić, Ines, Cook, James, Savić, Miroslav, Savić, Snežana, "Investigation of albumin adsorption on DK-I-56-1 nanocrystals by dynamic light scattering" in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija (2022),
https://hdl.handle.net/21.15107/rcub_farfar_4285 .

TY  - CONF
AU  - Mitrović, Jelena
AU  - Petković, Miloš
AU  - Ranđelović, Danijela
AU  - Đoković, Jelena
AU  - Knutson, Daniel
AU  - Cook, James
AU  - Savić, Vladimir
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4097
AB  - Lipid nanoparticles are being intensively investigated for
the formulation of the drugs with poor solubility substances
(1). They represent colloid dispersions of the particles with
lipid matrix that is solid at room and body temperature.
Because of the low capacity of triglycerides for the drug
substances incorporation, alternatively, high amounts of
lecithin could be added to increase the solubilization (2).
This was used for the incorporation of DK-I-60-3 (7-
methoxy-d3-2-(4-methoxyd3-phenyl)-2,5-dihydro-
3Hpyrazolo[4,3-c]quinolin-3-one), novel deuterated
pyrazoloquinolinone ligand, with very low solubility in
water as well as in oils (3,4). However, because of
amphiphilic nature of lecithin, its localization within
nanoparticles should be analyzed, especially with respect
to stability, drug loading capacity and drug localization,
because it may additionally influence the drug release
mechanism (2).
C3  - 13th World meeting on pharmaceutics, biopharmaceutics and pharmaceutical technology, 28-31 March 2022, Rotterdam, Netherlands
T1  - Physicochemical/structural investigation of lipid nanoparticles with high lecithin amounts loaded with patent protected pyrazoloquinolinone ligand DK-I-60-3
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4097
ER  - 

@conference{
author = "Mitrović, Jelena and Petković, Miloš and Ranđelović, Danijela and Đoković, Jelena and Knutson, Daniel and Cook, James and Savić, Vladimir and Savić, Miroslav and Savić, Snežana",
year = "2022",
abstract = "Lipid nanoparticles are being intensively investigated for
the formulation of the drugs with poor solubility substances
(1). They represent colloid dispersions of the particles with
lipid matrix that is solid at room and body temperature.
Because of the low capacity of triglycerides for the drug
substances incorporation, alternatively, high amounts of
lecithin could be added to increase the solubilization (2).
This was used for the incorporation of DK-I-60-3 (7-
methoxy-d3-2-(4-methoxyd3-phenyl)-2,5-dihydro-
3Hpyrazolo[4,3-c]quinolin-3-one), novel deuterated
pyrazoloquinolinone ligand, with very low solubility in
water as well as in oils (3,4). However, because of
amphiphilic nature of lecithin, its localization within
nanoparticles should be analyzed, especially with respect
to stability, drug loading capacity and drug localization,
because it may additionally influence the drug release
mechanism (2).",
journal = "13th World meeting on pharmaceutics, biopharmaceutics and pharmaceutical technology, 28-31 March 2022, Rotterdam, Netherlands",
title = "Physicochemical/structural investigation of lipid nanoparticles with high lecithin amounts loaded with patent protected pyrazoloquinolinone ligand DK-I-60-3",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4097"
}

Mitrović, J., Petković, M., Ranđelović, D., Đoković, J., Knutson, D., Cook, J., Savić, V., Savić, M.,& Savić, S.. (2022). Physicochemical/structural investigation of lipid nanoparticles with high lecithin amounts loaded with patent protected pyrazoloquinolinone ligand DK-I-60-3. in 13th World meeting on pharmaceutics, biopharmaceutics and pharmaceutical technology, 28-31 March 2022, Rotterdam, Netherlands.
https://hdl.handle.net/21.15107/rcub_farfar_4097

Mitrović J, Petković M, Ranđelović D, Đoković J, Knutson D, Cook J, Savić V, Savić M, Savić S. Physicochemical/structural investigation of lipid nanoparticles with high lecithin amounts loaded with patent protected pyrazoloquinolinone ligand DK-I-60-3. in 13th World meeting on pharmaceutics, biopharmaceutics and pharmaceutical technology, 28-31 March 2022, Rotterdam, Netherlands. 2022;.
https://hdl.handle.net/21.15107/rcub_farfar_4097 .

Mitrović, Jelena, Petković, Miloš, Ranđelović, Danijela, Đoković, Jelena, Knutson, Daniel, Cook, James, Savić, Vladimir, Savić, Miroslav, Savić, Snežana, "Physicochemical/structural investigation of lipid nanoparticles with high lecithin amounts loaded with patent protected pyrazoloquinolinone ligand DK-I-60-3" in 13th World meeting on pharmaceutics, biopharmaceutics and pharmaceutical technology, 28-31 March 2022, Rotterdam, Netherlands (2022),
https://hdl.handle.net/21.15107/rcub_farfar_4097 .

TY  - CONF
AU  - Stanković, Dragana
AU  - Janković, Drina
AU  - Mirković, Marija
AU  - Radović, Magdalena
AU  - Milanović, Zorana
AU  - Vukadinović, Aleksandar
AU  - Stanković, Aljoša
AU  - Perić, Marko
AU  - Vranješ Đurić, Sanja
AU  - Prijović, Željko
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4576
AB  - Nanoparticle delivery to solid tumors after an intravenous injection has shown to be
very limited in its ability to achieve therapeutic dosage in the tumor due to nonspecific
nanoparticle uptake by RES. To overcome these problems, local intratumoral injection of
nanoparticles is being investigated as more relevant route of administration. In the present
study, superparamagnetic iron oxide nanoparticles (SPIONs) were synthetized, coated with
citric (CA) or dimercaptosuccinic acid (DMSA) and radiolabeled with 90 Y or 177Lu, aiming to
develop radioactive nanoparticles for localized tumor therapy. Biodistribution and
antitumor efficacy of radiolabeled SPIONs after local intratumoral administration in CT26 or
4T1 xenografts-bearing BALB/c mice were studied. Tracking the radioactivity distribution of
injected 90 Y-CA-SPIONs and 177Lu-DMSA-SPIONs revealed that due to the size of the
nanoparticles, their diffusive escape from the tumor into healthy organs and tissues is
slowed down; the particles remain at the injection site up to 14 days after the injection, and
thereby increasing the tumor's exposure to radiation. Lower therapeutic efficacy of 177Lu-
DMSA-SPIONs in CT26 or 4T1 tumor can be explained by slight diffusion of particles from
injection sites into distant tumor regions and moderate-energy β-particles emitted by 177 Lu
(0.5MeV). These studies suggest that 90Y-CA-SPIONs is superior to 177 Lu-DMSA-SPIONs at
inhibiting both tumors growth, due to the high-energy β-particles emitted by 90 Y (2.27MeV)
and a longer path length. 90 Y is therapeutically superior to 177Lu in investigated xenograft
models. We believe that an intratumorally injected radiolabeled SPIONs can be considered as
a potential therapeutic agent for localized cancer therapy.
AB  - Prethodna istraživanja su pokazala da se intravenskim načinom aplikacije nanočestica
ne postiže zadovoljavajuća terapijska doza u solidnim tumorima, zbog nespecifičnog
preuzimanja nanočestica od strane retikuloendotelnog sistema. Da bi se prevazišli ovi
problemi, smatra se da je intratumorski način aplikacije pogodniji način primene nanočestica
u terapiji solidnih tumora. Sa ciljem da se razvije radiofarmaceutik za lokalizovanu terapiju
tumora, u ovim ispitivanjima, superparamagnetne nanočestice oksida gvožđa (SPION) su
sintetisane, površinski obložene limunskom (CA) i dimerkaptoćilibarnom (DMSA) kiselinom
i radioobeležene sa 90 Y i 177 Lu. Posebna pažnja je posvećena ispitivanjima distribucije i
antitumorske efikasnosti radioaktivno obeleženih SPIONa nakon lokalne intratumorske
primene u ksenografte indukovane supkutanim injekcijama CT26 i 4T1 ćelija BALB/c
miševima. Praćenje distribucije intratumorski injektovanih 90 Y-CA-SPION-a i 177 Lu-DMSA-
SPION-a je pokazalo da je zbog veličine nanočestica njihova migracija iz tumorskog tkiva u
zdrave organe i tkiva usporena, pa čestice ostaju na mestu ubrizgavanja do 14 dana, čime se
značajno povećava izloženost tumora zračenju. Niža terapijska efikasnost 177Lu-DMSA-
SPION-a u CT26 ili 4T1 tumorima se može objasniti slabom migracijom čestica sa mesta
aplikacije do udaljenih tumorskih ćelija kao i kratkim dometom u tkivu β– čestica koje
emituje 177 Lu zbog energije zračenja od 0,5MeV. Ova ispitivanja su pokazala da je 90 Y-CA-
SPION značajno efikasniji od 177 Lu-DMSA-SPION u inhibiciji rasta obe vrste tumora, zbog
visokoenergetskih β– čestica koje emituje 90 Y (2,27MeV) i većeg dometa u tkivu. 90 Y je
terapeutski superiorniji od 177 Lu u istraživanim modelima ksenografta. Mišljenja smo da se
intratumorski primenjeni radioaktivno obeleženi SPION-i mogu smatrati potencijalnim
terapijskim agensom za lokalizovanu terapiju solidnih, inoperabilnih i teško dostupnih
tumora.
PB  - Savez farmaceutskih udruženja Srbije (SFUS)
C3  - Arhiv za farmaciju
T1  - Efficacy of 177Lu- and 90Y-labeled nanoparticles in targeted tumor therapy in a mouse CT26 and 4T1 xenograft model
T1  - Efikasnost 177Lu‐ i 90 Y‐obeleženih nanočestica u ciljanoj terapiji tumora na modelu mišjih CT26 I 4T1 ksenografta
VL  - 72
IS  - 4 suplement
SP  - S428
EP  - S429
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4576
ER  - 

@conference{
author = "Stanković, Dragana and Janković, Drina and Mirković, Marija and Radović, Magdalena and Milanović, Zorana and Vukadinović, Aleksandar and Stanković, Aljoša and Perić, Marko and Vranješ Đurić, Sanja and Prijović, Željko and Savić, Miroslav",
year = "2022",
abstract = "Nanoparticle delivery to solid tumors after an intravenous injection has shown to be
very limited in its ability to achieve therapeutic dosage in the tumor due to nonspecific
nanoparticle uptake by RES. To overcome these problems, local intratumoral injection of
nanoparticles is being investigated as more relevant route of administration. In the present
study, superparamagnetic iron oxide nanoparticles (SPIONs) were synthetized, coated with
citric (CA) or dimercaptosuccinic acid (DMSA) and radiolabeled with 90 Y or 177Lu, aiming to
develop radioactive nanoparticles for localized tumor therapy. Biodistribution and
antitumor efficacy of radiolabeled SPIONs after local intratumoral administration in CT26 or
4T1 xenografts-bearing BALB/c mice were studied. Tracking the radioactivity distribution of
injected 90 Y-CA-SPIONs and 177Lu-DMSA-SPIONs revealed that due to the size of the
nanoparticles, their diffusive escape from the tumor into healthy organs and tissues is
slowed down; the particles remain at the injection site up to 14 days after the injection, and
thereby increasing the tumor's exposure to radiation. Lower therapeutic efficacy of 177Lu-
DMSA-SPIONs in CT26 or 4T1 tumor can be explained by slight diffusion of particles from
injection sites into distant tumor regions and moderate-energy β-particles emitted by 177 Lu
(0.5MeV). These studies suggest that 90Y-CA-SPIONs is superior to 177 Lu-DMSA-SPIONs at
inhibiting both tumors growth, due to the high-energy β-particles emitted by 90 Y (2.27MeV)
and a longer path length. 90 Y is therapeutically superior to 177Lu in investigated xenograft
models. We believe that an intratumorally injected radiolabeled SPIONs can be considered as
a potential therapeutic agent for localized cancer therapy., Prethodna istraživanja su pokazala da se intravenskim načinom aplikacije nanočestica
ne postiže zadovoljavajuća terapijska doza u solidnim tumorima, zbog nespecifičnog
preuzimanja nanočestica od strane retikuloendotelnog sistema. Da bi se prevazišli ovi
problemi, smatra se da je intratumorski način aplikacije pogodniji način primene nanočestica
u terapiji solidnih tumora. Sa ciljem da se razvije radiofarmaceutik za lokalizovanu terapiju
tumora, u ovim ispitivanjima, superparamagnetne nanočestice oksida gvožđa (SPION) su
sintetisane, površinski obložene limunskom (CA) i dimerkaptoćilibarnom (DMSA) kiselinom
i radioobeležene sa 90 Y i 177 Lu. Posebna pažnja je posvećena ispitivanjima distribucije i
antitumorske efikasnosti radioaktivno obeleženih SPIONa nakon lokalne intratumorske
primene u ksenografte indukovane supkutanim injekcijama CT26 i 4T1 ćelija BALB/c
miševima. Praćenje distribucije intratumorski injektovanih 90 Y-CA-SPION-a i 177 Lu-DMSA-
SPION-a je pokazalo da je zbog veličine nanočestica njihova migracija iz tumorskog tkiva u
zdrave organe i tkiva usporena, pa čestice ostaju na mestu ubrizgavanja do 14 dana, čime se
značajno povećava izloženost tumora zračenju. Niža terapijska efikasnost 177Lu-DMSA-
SPION-a u CT26 ili 4T1 tumorima se može objasniti slabom migracijom čestica sa mesta
aplikacije do udaljenih tumorskih ćelija kao i kratkim dometom u tkivu β– čestica koje
emituje 177 Lu zbog energije zračenja od 0,5MeV. Ova ispitivanja su pokazala da je 90 Y-CA-
SPION značajno efikasniji od 177 Lu-DMSA-SPION u inhibiciji rasta obe vrste tumora, zbog
visokoenergetskih β– čestica koje emituje 90 Y (2,27MeV) i većeg dometa u tkivu. 90 Y je
terapeutski superiorniji od 177 Lu u istraživanim modelima ksenografta. Mišljenja smo da se
intratumorski primenjeni radioaktivno obeleženi SPION-i mogu smatrati potencijalnim
terapijskim agensom za lokalizovanu terapiju solidnih, inoperabilnih i teško dostupnih
tumora.",
publisher = "Savez farmaceutskih udruženja Srbije (SFUS)",
journal = "Arhiv za farmaciju",
title = "Efficacy of 177Lu- and 90Y-labeled nanoparticles in targeted tumor therapy in a mouse CT26 and 4T1 xenograft model, Efikasnost 177Lu‐ i 90 Y‐obeleženih nanočestica u ciljanoj terapiji tumora na modelu mišjih CT26 I 4T1 ksenografta",
volume = "72",
number = "4 suplement",
pages = "S428-S429",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4576"
}

Stanković, D., Janković, D., Mirković, M., Radović, M., Milanović, Z., Vukadinović, A., Stanković, A., Perić, M., Vranješ Đurić, S., Prijović, Ž.,& Savić, M.. (2022). Efficacy of 177Lu- and 90Y-labeled nanoparticles in targeted tumor therapy in a mouse CT26 and 4T1 xenograft model. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije (SFUS)., 72(4 suplement), S428-S429.
https://hdl.handle.net/21.15107/rcub_farfar_4576

Stanković D, Janković D, Mirković M, Radović M, Milanović Z, Vukadinović A, Stanković A, Perić M, Vranješ Đurić S, Prijović Ž, Savić M. Efficacy of 177Lu- and 90Y-labeled nanoparticles in targeted tumor therapy in a mouse CT26 and 4T1 xenograft model. in Arhiv za farmaciju. 2022;72(4 suplement):S428-S429.
https://hdl.handle.net/21.15107/rcub_farfar_4576 .

Stanković, Dragana, Janković, Drina, Mirković, Marija, Radović, Magdalena, Milanović, Zorana, Vukadinović, Aleksandar, Stanković, Aljoša, Perić, Marko, Vranješ Đurić, Sanja, Prijović, Željko, Savić, Miroslav, "Efficacy of 177Lu- and 90Y-labeled nanoparticles in targeted tumor therapy in a mouse CT26 and 4T1 xenograft model" in Arhiv za farmaciju, 72, no. 4 suplement (2022):S428-S429,
https://hdl.handle.net/21.15107/rcub_farfar_4576 .

TY  - CONF
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4483
AB  - The adverse effects of pharmaceuticals on the central or peripheral nervous systems
are poorly predicted by the current in vitro and in vivo preclinical studies performed during
research and development process. Increasing the predictivity of the preclinical toolbox is a
clear need, and would benefit to human volunteers/patients (safer drugs) and
pharmaceutical industry (reduced attrition). The NeuroDeRisk Consortium consists of 18
academic, pharmaceutical industry and small and medium enterprise partners within the
Innovative Medicines Initiative project (2019-2022). University of Belgrade – Faculty of
Pharmacy is the partner that leads the work package devoted to preclinical prediction of
mood and cognitive adverse effects. The other two groups of challenging adverse effects
tackled by the project are seizures and peripheral neuropathies. By employing a plethora of
experimental techniques and numerous pharmaceuticals previously connected with each of
major adverse effects examined, the project looks for innovative tools, assays and protocols,
which cover in silico, in vitro, in vivo and ex vivo approaches. Besides widening the
knowledge on many widely used pharmaceuticals, the major goal of the project is to develop
an integrated platform for better risk-assessment in exploratory and regulatory studies in
the future.
AB  - Postojeće in vitro i in vivo prekliničke studije koje se sprovode tokom procesa
istraživanja i razvoja teško mogu predvideti neželjene efekte farmaceutika na centralni i
periferni nervni sistem. Postoji jasna potreba za povećanjem prediktivnosti prekliničkih
oruđa, što bi bilo korisno za humane volontere/pacijente (bezbedniji lekovi) i farmaceutsku
industriju (smanjena stopa neuspeha). NeuroDeRisk konzorcijum se sastoji od 18
akademskih institucija, kompanija farmaceutske industrije i malih i srednjih preduzeća u
okviru projekta Inicijative za inovativne lekove (2019-2022). Univerzitet u Beogradu –
Farmaceutski fakultet jeste partner koji predvodi radni parket posvećen prekliničkom
predviđanju neželjenih efekata na raspoloženje i kogniciju. Druge dve grupe izazovnih
neželjenih efekata koje projekat obrađuje jesu konvulzivni napadi i periferne neuropatije.
Korišćenjem mnoštva eksperimentalnih tehnika i brojnih farmaceutika koji su prethodno
povezani sa svakim od ispitivanih velikih neželjenih efekata, projekat traga za inovativnim
oruđima, esejima i protokolima, koji pokrivaju in silico, in vitro, in vivo i ex vivo pristupe.
Mimo širenja znanja o brojnim široko korišćenim farmaceuticima, glavni cilj projekta jeste
razvoj integrisane platforme za bolju procenu rizika u budućim eksploratornim i
regulatornim studijama.
PB  - Savez farmaceutskih udruženja Srbije (SFUS)
C3  - Arhiv za farmaciju
T1  - Neurotoxicity De-Risking in Preclinical Drug Discovery
T1  - Prekliničko predviđanje neželjenih efekata lekova na nervni sistem
VL  - 72
IS  - 4 suplement
SP  - S163
EP  - S164
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4483
ER  - 

@conference{
author = "Savić, Miroslav",
year = "2022",
abstract = "The adverse effects of pharmaceuticals on the central or peripheral nervous systems
are poorly predicted by the current in vitro and in vivo preclinical studies performed during
research and development process. Increasing the predictivity of the preclinical toolbox is a
clear need, and would benefit to human volunteers/patients (safer drugs) and
pharmaceutical industry (reduced attrition). The NeuroDeRisk Consortium consists of 18
academic, pharmaceutical industry and small and medium enterprise partners within the
Innovative Medicines Initiative project (2019-2022). University of Belgrade – Faculty of
Pharmacy is the partner that leads the work package devoted to preclinical prediction of
mood and cognitive adverse effects. The other two groups of challenging adverse effects
tackled by the project are seizures and peripheral neuropathies. By employing a plethora of
experimental techniques and numerous pharmaceuticals previously connected with each of
major adverse effects examined, the project looks for innovative tools, assays and protocols,
which cover in silico, in vitro, in vivo and ex vivo approaches. Besides widening the
knowledge on many widely used pharmaceuticals, the major goal of the project is to develop
an integrated platform for better risk-assessment in exploratory and regulatory studies in
the future., Postojeće in vitro i in vivo prekliničke studije koje se sprovode tokom procesa
istraživanja i razvoja teško mogu predvideti neželjene efekte farmaceutika na centralni i
periferni nervni sistem. Postoji jasna potreba za povećanjem prediktivnosti prekliničkih
oruđa, što bi bilo korisno za humane volontere/pacijente (bezbedniji lekovi) i farmaceutsku
industriju (smanjena stopa neuspeha). NeuroDeRisk konzorcijum se sastoji od 18
akademskih institucija, kompanija farmaceutske industrije i malih i srednjih preduzeća u
okviru projekta Inicijative za inovativne lekove (2019-2022). Univerzitet u Beogradu –
Farmaceutski fakultet jeste partner koji predvodi radni parket posvećen prekliničkom
predviđanju neželjenih efekata na raspoloženje i kogniciju. Druge dve grupe izazovnih
neželjenih efekata koje projekat obrađuje jesu konvulzivni napadi i periferne neuropatije.
Korišćenjem mnoštva eksperimentalnih tehnika i brojnih farmaceutika koji su prethodno
povezani sa svakim od ispitivanih velikih neželjenih efekata, projekat traga za inovativnim
oruđima, esejima i protokolima, koji pokrivaju in silico, in vitro, in vivo i ex vivo pristupe.
Mimo širenja znanja o brojnim široko korišćenim farmaceuticima, glavni cilj projekta jeste
razvoj integrisane platforme za bolju procenu rizika u budućim eksploratornim i
regulatornim studijama.",
publisher = "Savez farmaceutskih udruženja Srbije (SFUS)",
journal = "Arhiv za farmaciju",
title = "Neurotoxicity De-Risking in Preclinical Drug Discovery, Prekliničko predviđanje neželjenih efekata lekova na nervni sistem",
volume = "72",
number = "4 suplement",
pages = "S163-S164",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4483"
}

Savić, M.. (2022). Neurotoxicity De-Risking in Preclinical Drug Discovery. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije (SFUS)., 72(4 suplement), S163-S164.
https://hdl.handle.net/21.15107/rcub_farfar_4483

Savić M. Neurotoxicity De-Risking in Preclinical Drug Discovery. in Arhiv za farmaciju. 2022;72(4 suplement):S163-S164.
https://hdl.handle.net/21.15107/rcub_farfar_4483 .

Savić, Miroslav, "Neurotoxicity De-Risking in Preclinical Drug Discovery" in Arhiv za farmaciju, 72, no. 4 suplement (2022):S163-S164,
https://hdl.handle.net/21.15107/rcub_farfar_4483 .

TY  - CONF
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4466
AB  - Throughout the process of drug discovery, development and commercialization,
impact of adverse effects on nervous system is second only to cardiovascular adverse effects.
Many of these effects are within the interconnected domains of mood and cognition, usually
with low incidence and thus more difficult to be revealed. While most of them have more or
less impact on quality of life, some are life-threating, directly (suicide ideation and behavior,
including completed suicide) or indirectly (cognitive impairment that affects driving
performance). Intriguingly, mood and cognitive adverse effects (MCAEs) are encountered
not only with low-molecular weight pharmaceuticals that readily pass the blood-brain
barrier, but also with biological drugs, most notably several monoclonal antibodies. Besides
a number of receptors, transporters and other elements involved in processes of
neurotransmission and neuronal plasticity, neuroimmunomodulation has taken a distinct
place in still limited understanding the mechanisms that induce MCAEs. The latter concept
corresponds with substantial data demonstrating that inflammatory signals generated both
in the central nervous system and in the peripheral tissues play an important role in
pathogenesis of various psychiatric and neurological disorders. It is possible that changes in
conventional immune components, such are interleukins, are linked with impaired
behavioral manifestations through metabolites of the kynurenine pathway and/or changes
in activity of non-neuronal glial cells. In clinical as well as experimental settings, recognition
of MCAEs is hindered by difficulties in differentiation between normal compared to
pathological changes in behavior – especially having in mind substantial variations in output
of behavioral parameters both in humans and experimental animals.
AB  - Tokom procesa otkrića, razvoja i komercijalizacije lekova, jedino su kardiovaskularni
neželjeni efekti češći nego oni koji su vezani za nervni sistem. Mnogi od ovih efekata se
nalaze u povezanim domenima raspoloženja i kognicije, često sa niskom učestalošću
javljanja, što otežava njihovo prepoznavanje. Dok većina ovih efekata ima veći ili manji uticaj
na kvalitet života, pojedini među njima i ugrožavaju život, direktno (suicidalna ideacija i
ponašanje, uključujući izvršeni suicid) ili indirektno (kognitivno oštećenje koje utiče na
sposobnost vožnje). Intrigantno, neželjeni efekti na raspoloženje i kogniciju (NERK) sreću se
ne samo sa farmaceuticima male molekulske mase, koji lako prolaze krvno-moždanu
barijeru, nego i sa biološkim lekovima, najistaknutije sa nekoliko monoklonskih antitela.
Pored pojedinih receptora, transportera i drugih elemenata uključenih u procese
neurotransmisije i neuronske plastičnosti, neuroimunomodulacija zauzima istaknuto mesto
u još uvek ograničenom razumevanju mehanizama koji dovode do NERK. Ovaj poslednji
koncept korespondira sa obimnim nalazima koji potvrđuju da u patogenezi različitih
psihijatrijskih i neuroloških poremećaja značajnu ulogu imaju inflamatorni signali koji se
generišu kako u centralnom nervnom sistemu, tako i u perifernim tkivima. Moguće je da
metaboliti kinureninskog puta i/ili promene u aktivnosti ne-neuronskih glijalnih ćelija
povezuju promene u konvencionalnim imunskim komponentama, kao što su interleukini, sa
manifestacijama oštećenog ponašanja. U kliničkim kao i u eksperimentalnim uslovima,
prepoznavanje NERK ograničavaju poteškoće u diferencijaciji između normalnih i patoloških
promena u ponašanju – posebno kada se uzmu u obzir znatne varijacije u vrednostima
parametara ponašanja kako kod ljudi tako i kod eksperimentalnih životinja.
PB  - Savez farmaceutskih udruženja Srbije (SFUS)
C3  - Arhiv za farmaciju
T1  - Adverse effects of pharmaceuticals on mood, cognition and behavior
T1  - Neželjena dejstva lekova na raspoloženje, kogniciju i ponašanje
VL  - 72
IS  - 4 suplement
SP  - S122
EP  - S123
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4466
ER  - 

@conference{
author = "Savić, Miroslav",
year = "2022",
abstract = "Throughout the process of drug discovery, development and commercialization,
impact of adverse effects on nervous system is second only to cardiovascular adverse effects.
Many of these effects are within the interconnected domains of mood and cognition, usually
with low incidence and thus more difficult to be revealed. While most of them have more or
less impact on quality of life, some are life-threating, directly (suicide ideation and behavior,
including completed suicide) or indirectly (cognitive impairment that affects driving
performance). Intriguingly, mood and cognitive adverse effects (MCAEs) are encountered
not only with low-molecular weight pharmaceuticals that readily pass the blood-brain
barrier, but also with biological drugs, most notably several monoclonal antibodies. Besides
a number of receptors, transporters and other elements involved in processes of
neurotransmission and neuronal plasticity, neuroimmunomodulation has taken a distinct
place in still limited understanding the mechanisms that induce MCAEs. The latter concept
corresponds with substantial data demonstrating that inflammatory signals generated both
in the central nervous system and in the peripheral tissues play an important role in
pathogenesis of various psychiatric and neurological disorders. It is possible that changes in
conventional immune components, such are interleukins, are linked with impaired
behavioral manifestations through metabolites of the kynurenine pathway and/or changes
in activity of non-neuronal glial cells. In clinical as well as experimental settings, recognition
of MCAEs is hindered by difficulties in differentiation between normal compared to
pathological changes in behavior – especially having in mind substantial variations in output
of behavioral parameters both in humans and experimental animals., Tokom procesa otkrića, razvoja i komercijalizacije lekova, jedino su kardiovaskularni
neželjeni efekti češći nego oni koji su vezani za nervni sistem. Mnogi od ovih efekata se
nalaze u povezanim domenima raspoloženja i kognicije, često sa niskom učestalošću
javljanja, što otežava njihovo prepoznavanje. Dok većina ovih efekata ima veći ili manji uticaj
na kvalitet života, pojedini među njima i ugrožavaju život, direktno (suicidalna ideacija i
ponašanje, uključujući izvršeni suicid) ili indirektno (kognitivno oštećenje koje utiče na
sposobnost vožnje). Intrigantno, neželjeni efekti na raspoloženje i kogniciju (NERK) sreću se
ne samo sa farmaceuticima male molekulske mase, koji lako prolaze krvno-moždanu
barijeru, nego i sa biološkim lekovima, najistaknutije sa nekoliko monoklonskih antitela.
Pored pojedinih receptora, transportera i drugih elemenata uključenih u procese
neurotransmisije i neuronske plastičnosti, neuroimunomodulacija zauzima istaknuto mesto
u još uvek ograničenom razumevanju mehanizama koji dovode do NERK. Ovaj poslednji
koncept korespondira sa obimnim nalazima koji potvrđuju da u patogenezi različitih
psihijatrijskih i neuroloških poremećaja značajnu ulogu imaju inflamatorni signali koji se
generišu kako u centralnom nervnom sistemu, tako i u perifernim tkivima. Moguće je da
metaboliti kinureninskog puta i/ili promene u aktivnosti ne-neuronskih glijalnih ćelija
povezuju promene u konvencionalnim imunskim komponentama, kao što su interleukini, sa
manifestacijama oštećenog ponašanja. U kliničkim kao i u eksperimentalnim uslovima,
prepoznavanje NERK ograničavaju poteškoće u diferencijaciji između normalnih i patoloških
promena u ponašanju – posebno kada se uzmu u obzir znatne varijacije u vrednostima
parametara ponašanja kako kod ljudi tako i kod eksperimentalnih životinja.",
publisher = "Savez farmaceutskih udruženja Srbije (SFUS)",
journal = "Arhiv za farmaciju",
title = "Adverse effects of pharmaceuticals on mood, cognition and behavior, Neželjena dejstva lekova na raspoloženje, kogniciju i ponašanje",
volume = "72",
number = "4 suplement",
pages = "S122-S123",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4466"
}

Savić, M.. (2022). Adverse effects of pharmaceuticals on mood, cognition and behavior. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije (SFUS)., 72(4 suplement), S122-S123.
https://hdl.handle.net/21.15107/rcub_farfar_4466

Savić M. Adverse effects of pharmaceuticals on mood, cognition and behavior. in Arhiv za farmaciju. 2022;72(4 suplement):S122-S123.
https://hdl.handle.net/21.15107/rcub_farfar_4466 .

Savić, Miroslav, "Adverse effects of pharmaceuticals on mood, cognition and behavior" in Arhiv za farmaciju, 72, no. 4 suplement (2022):S122-S123,
https://hdl.handle.net/21.15107/rcub_farfar_4466 .

TY  - CONF
AU  - Savić, Snežana
AU  - Pantelić, Ivana
AU  - Jančić, Ivan
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4297
AB  - Mood, anxiety and cognitive symptoms in psychiatry and neurology represent a significant worldwide burden. Due to difficulties in disease modeling and drug delivering to the site of action, as well as gaps in in vitro/in vivo extrapolation, the efforts to elucidate the roles of stress and neuroimmune pathways in both, etiology and therapy of these symptoms are challenging, but may nevertheless result in novel mechanisms of action. Recent preclinical studies provided novel leads/drug candidates with promising mood, anxiety and cognitive effects, the intellectual property rights of which are co-owned by the project beneficiary. We aim to: (1) incorporate the selective ligands of GABAA and/or sigma-2 receptors, with code names GL-II-73, DK-I-56, MM-I-03 and CW-02-79, together with two reference sigma-2 receptor ligands (siramesine and RHM-1), into the optimized nanoparticles and target their delivery to the human induced pluripotent stem cell (hiPSC)- based tri-culture cell neuroinflammation model, or rat brain; (2) quantify the immunological/morphological/neurochemical markers in immunologically challenged hiPSC-derived neurons, astrocytes and glia cells, and (3) assess their effects on behavior and biological markers in immunologically challenged animals of both sexes subjected to chronic mild unpredictable stress. We assume that the targeted nanodelivery of selected compounds to the brain will improve their pharmacokinetic profile, fortify their beneficial effect on mood, anxiety and cognition, and help delineate the contributing neuroimmune effects presumably arising mainly from microglia. The familiarization with neuroimmune aspects and pharmacokinetic optimization will support the preclinical progress of these compounds and might provide a rationale for designing clinical trials.
AB  - Raspoloženje, anksioznost i kognitivni simptomi u psihijatriji i neurologiji predstavljaju
značajno opterećenje za zdravstvene sisteme na globalnom nivou. Usled poteškoća u
modelovanju bolesti i isporuci lekova na mesto delovanja, kao i jaza u in vitro/in vivo
ekstrapolaciji, potreba da se osvetle uloge stresa i neuroimunskih puteva u etiologiji i terapiji
ovih simptoma je izazov, i može rezultovati u novim mehanizmima delovanja. Nedavne
prekliničke studije obezbedile su nova vodeća jedinjenja/lekove kandidate sa obećavajućim
efektima na raspoloženje, anksioznost i kogniciju, rezultujući prihvatanjem patentnih prava
čiji je suvlasnik predlagač projekta. Cilj projekta je da: (1) inkorporiramo ligande selektivne
za GABAA i/ili sigma-2 receptore, sa kodiranim imenima GL-II-73, DK-I56, MM-I-03 i
CW-02-79, zajedno sa dva referentna liganda za sigma-2 receptore (siramesin i RHM-1), u
optimizovane nanočestice i da omogućimo njihovu ciljnu isporuku u hiPSCbazirani trićelijski
model neuroinflamacije, ili mozak pacova; (2) kvantifikujemo
imunološke/morfološke/neurohemijske markere u imunološki izazvanim hiPSC-derivisanim
neuronima, astrocitima i glija ćelijama, i (3) procenimo njihove efekte na ponašanje i
biološke markere u imunološki izazvanim životinjama oba pola podvrgnutim blagom
neočekivanom stresu. Procenjujemo da će ciljana isporuka odabranih jedinjenja pomoću
nanonosača poboljšati njihove farmakokinetičke (FK) profile, ojačati njihove korisne efekte
na raspoloženje, anksioznost i kogniciju i pomoći delineaciji doprinosa neuroimunskih
efekata, po svemu sudeći, poreklom uglavnom od mikroglija ćelija. Upoznavanje sa
neuroimunskim aspektima i FK optimizacija mogu da podrže napredak u prekliničkom
razvoju ovih jedinjenja i obezbede osnov za dizajniranje prospektivnih kliničkih studija.
C3  - 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija
T1  - Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at gabaa and/or sigma‐2 receptors: in vitro/in vivo delineation by nano‐ and hipsc‐based platforms
T1  - Neuroimunski aspekti efekata vodećih jedinjenja/lekova kandidata koji deluju preko gabaa i/ili sigma‐2 receptora na raspoloženje, anksioznost i kogniciju: in vitro/in vivo delineacija primenom nano‐ i hipsc platformi
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4297
ER  - 

@conference{
author = "Savić, Snežana and Pantelić, Ivana and Jančić, Ivan and Savić, Miroslav",
year = "2022",
abstract = "Mood, anxiety and cognitive symptoms in psychiatry and neurology represent a significant worldwide burden. Due to difficulties in disease modeling and drug delivering to the site of action, as well as gaps in in vitro/in vivo extrapolation, the efforts to elucidate the roles of stress and neuroimmune pathways in both, etiology and therapy of these symptoms are challenging, but may nevertheless result in novel mechanisms of action. Recent preclinical studies provided novel leads/drug candidates with promising mood, anxiety and cognitive effects, the intellectual property rights of which are co-owned by the project beneficiary. We aim to: (1) incorporate the selective ligands of GABAA and/or sigma-2 receptors, with code names GL-II-73, DK-I-56, MM-I-03 and CW-02-79, together with two reference sigma-2 receptor ligands (siramesine and RHM-1), into the optimized nanoparticles and target their delivery to the human induced pluripotent stem cell (hiPSC)- based tri-culture cell neuroinflammation model, or rat brain; (2) quantify the immunological/morphological/neurochemical markers in immunologically challenged hiPSC-derived neurons, astrocytes and glia cells, and (3) assess their effects on behavior and biological markers in immunologically challenged animals of both sexes subjected to chronic mild unpredictable stress. We assume that the targeted nanodelivery of selected compounds to the brain will improve their pharmacokinetic profile, fortify their beneficial effect on mood, anxiety and cognition, and help delineate the contributing neuroimmune effects presumably arising mainly from microglia. The familiarization with neuroimmune aspects and pharmacokinetic optimization will support the preclinical progress of these compounds and might provide a rationale for designing clinical trials., Raspoloženje, anksioznost i kognitivni simptomi u psihijatriji i neurologiji predstavljaju
značajno opterećenje za zdravstvene sisteme na globalnom nivou. Usled poteškoća u
modelovanju bolesti i isporuci lekova na mesto delovanja, kao i jaza u in vitro/in vivo
ekstrapolaciji, potreba da se osvetle uloge stresa i neuroimunskih puteva u etiologiji i terapiji
ovih simptoma je izazov, i može rezultovati u novim mehanizmima delovanja. Nedavne
prekliničke studije obezbedile su nova vodeća jedinjenja/lekove kandidate sa obećavajućim
efektima na raspoloženje, anksioznost i kogniciju, rezultujući prihvatanjem patentnih prava
čiji je suvlasnik predlagač projekta. Cilj projekta je da: (1) inkorporiramo ligande selektivne
za GABAA i/ili sigma-2 receptore, sa kodiranim imenima GL-II-73, DK-I56, MM-I-03 i
CW-02-79, zajedno sa dva referentna liganda za sigma-2 receptore (siramesin i RHM-1), u
optimizovane nanočestice i da omogućimo njihovu ciljnu isporuku u hiPSCbazirani trićelijski
model neuroinflamacije, ili mozak pacova; (2) kvantifikujemo
imunološke/morfološke/neurohemijske markere u imunološki izazvanim hiPSC-derivisanim
neuronima, astrocitima i glija ćelijama, i (3) procenimo njihove efekte na ponašanje i
biološke markere u imunološki izazvanim životinjama oba pola podvrgnutim blagom
neočekivanom stresu. Procenjujemo da će ciljana isporuka odabranih jedinjenja pomoću
nanonosača poboljšati njihove farmakokinetičke (FK) profile, ojačati njihove korisne efekte
na raspoloženje, anksioznost i kogniciju i pomoći delineaciji doprinosa neuroimunskih
efekata, po svemu sudeći, poreklom uglavnom od mikroglija ćelija. Upoznavanje sa
neuroimunskim aspektima i FK optimizacija mogu da podrže napredak u prekliničkom
razvoju ovih jedinjenja i obezbede osnov za dizajniranje prospektivnih kliničkih studija.",
journal = "8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija",
title = "Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at gabaa and/or sigma‐2 receptors: in vitro/in vivo delineation by nano‐ and hipsc‐based platforms, Neuroimunski aspekti efekata vodećih jedinjenja/lekova kandidata koji deluju preko gabaa i/ili sigma‐2 receptora na raspoloženje, anksioznost i kogniciju: in vitro/in vivo delineacija primenom nano‐ i hipsc platformi",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4297"
}

Savić, S., Pantelić, I., Jančić, I.,& Savić, M.. (2022). Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at gabaa and/or sigma‐2 receptors: in vitro/in vivo delineation by nano‐ and hipsc‐based platforms. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija.
https://hdl.handle.net/21.15107/rcub_farfar_4297

Savić S, Pantelić I, Jančić I, Savić M. Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at gabaa and/or sigma‐2 receptors: in vitro/in vivo delineation by nano‐ and hipsc‐based platforms. in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija. 2022;.
https://hdl.handle.net/21.15107/rcub_farfar_4297 .

Savić, Snežana, Pantelić, Ivana, Jančić, Ivan, Savić, Miroslav, "Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at gabaa and/or sigma‐2 receptors: in vitro/in vivo delineation by nano‐ and hipsc‐based platforms" in 8. Kongres farmaceuta Srbije; 12.-15. oktobar, Beograd, Srbija (2022),
https://hdl.handle.net/21.15107/rcub_farfar_4297 .

TY  - JOUR
AU  - Divović-Matović, Branka
AU  - Knutson, Dan
AU  - Mitrović, Jelena
AU  - Stevanović, Vladimir
AU  - Stanojević, Boban
AU  - Savić, Snežana
AU  - Cook, James
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4289
AB  - Several pyrazoloquinolinone (PQ) ligands were recently discovered as func-tionally selective positive modulators at the PQ site ofα6-containing GABAAreceptors. PQs are also neutral modulators at the benzodiazepine site. Weassessed the influence of PQ compounds from three structural groups (PZ-II-029 and related deuterated analogues DK-I-56-1, RV-I-029, DK-I-60-3 and DK-I-86-1; LAU 463 and related analogues DK-I-58-1 and DK-II-58-1; and DK-I-87-1), alone and in combination with diazepam, on the behaviour of maleSprague–Dawley rats. An excellent behavioural safety profile of all tested PQswas demonstrated in the spontaneous locomotor activity, rotarod, loss of right-ing reflex and pentylenetetrazol tests. In interaction studies, only PZ-II-029and its analogues prevented the ataxic effects of the benzodiazepine, asassessed in the rotarod test and during monitoring of rat locomotor activityafter awakening from the loss of righting reflex. Published electrophysiologicalprofiles of PQ ligands imply that positive modulation elicited atα6-GABAAreceptors that contain theγ2 andδsubunit, rather than their neutral modula-tory action at the benzodiazepine site, may prevent the ataxic action of diaze-pam. Thus, PZ-II-029 and its deuterated analogues are not prone to untowardinteractions with benzodiazepines and may indeed completely abolish theirataxic action, seen at therapeutic, and especially toxic concentrations.
PB  - John Wiley and Sons Inc
T2  - Basic and Clinical Pharmacology and Toxicology
T1  - Behavioural interaction of pyrazoloquinolinone positive allosteric modulators at α6GABAA receptors and diazepam in rats: Anti-diazepam-induced ataxia action as a structure-dependent feature
VL  - 131
IS  - 6
SP  - 514
EP  - 524
DO  - 10.1111/bcpt.13801
ER  - 

@article{
author = "Divović-Matović, Branka and Knutson, Dan and Mitrović, Jelena and Stevanović, Vladimir and Stanojević, Boban and Savić, Snežana and Cook, James and Savić, Miroslav",
year = "2022",
abstract = "Several pyrazoloquinolinone (PQ) ligands were recently discovered as func-tionally selective positive modulators at the PQ site ofα6-containing GABAAreceptors. PQs are also neutral modulators at the benzodiazepine site. Weassessed the influence of PQ compounds from three structural groups (PZ-II-029 and related deuterated analogues DK-I-56-1, RV-I-029, DK-I-60-3 and DK-I-86-1; LAU 463 and related analogues DK-I-58-1 and DK-II-58-1; and DK-I-87-1), alone and in combination with diazepam, on the behaviour of maleSprague–Dawley rats. An excellent behavioural safety profile of all tested PQswas demonstrated in the spontaneous locomotor activity, rotarod, loss of right-ing reflex and pentylenetetrazol tests. In interaction studies, only PZ-II-029and its analogues prevented the ataxic effects of the benzodiazepine, asassessed in the rotarod test and during monitoring of rat locomotor activityafter awakening from the loss of righting reflex. Published electrophysiologicalprofiles of PQ ligands imply that positive modulation elicited atα6-GABAAreceptors that contain theγ2 andδsubunit, rather than their neutral modula-tory action at the benzodiazepine site, may prevent the ataxic action of diaze-pam. Thus, PZ-II-029 and its deuterated analogues are not prone to untowardinteractions with benzodiazepines and may indeed completely abolish theirataxic action, seen at therapeutic, and especially toxic concentrations.",
publisher = "John Wiley and Sons Inc",
journal = "Basic and Clinical Pharmacology and Toxicology",
title = "Behavioural interaction of pyrazoloquinolinone positive allosteric modulators at α6GABAA receptors and diazepam in rats: Anti-diazepam-induced ataxia action as a structure-dependent feature",
volume = "131",
number = "6",
pages = "514-524",
doi = "10.1111/bcpt.13801"
}

Divović-Matović, B., Knutson, D., Mitrović, J., Stevanović, V., Stanojević, B., Savić, S., Cook, J.,& Savić, M.. (2022). Behavioural interaction of pyrazoloquinolinone positive allosteric modulators at α6GABAA receptors and diazepam in rats: Anti-diazepam-induced ataxia action as a structure-dependent feature. in Basic and Clinical Pharmacology and Toxicology
John Wiley and Sons Inc., 131(6), 514-524.
https://doi.org/10.1111/bcpt.13801

Divović-Matović B, Knutson D, Mitrović J, Stevanović V, Stanojević B, Savić S, Cook J, Savić M. Behavioural interaction of pyrazoloquinolinone positive allosteric modulators at α6GABAA receptors and diazepam in rats: Anti-diazepam-induced ataxia action as a structure-dependent feature. in Basic and Clinical Pharmacology and Toxicology. 2022;131(6):514-524.
doi:10.1111/bcpt.13801 .

Divović-Matović, Branka, Knutson, Dan, Mitrović, Jelena, Stevanović, Vladimir, Stanojević, Boban, Savić, Snežana, Cook, James, Savić, Miroslav, "Behavioural interaction of pyrazoloquinolinone positive allosteric modulators at α6GABAA receptors and diazepam in rats: Anti-diazepam-induced ataxia action as a structure-dependent feature" in Basic and Clinical Pharmacology and Toxicology, 131, no. 6 (2022):514-524,
https://doi.org/10.1111/bcpt.13801 . .

TY  - JOUR
AU  - Mian, Md Yeunus
AU  - Divović, Branka
AU  - Sharmin, Dishary
AU  - Pandey, Kamal P.
AU  - Golani, Lalit K.
AU  - Tiruveedhula, V. V. N. Phani Babu
AU  - Cerne, Rok
AU  - Smith, Jodi L.
AU  - Ping, Xingjie
AU  - Jin, Xiaoming
AU  - Imler, Gregory H.
AU  - Deschamps, Jeffrey R.
AU  - Lippa, Arnold
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Rowlett, James
AU  - Witkin, Jeffrey M.
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4363
AB  - Imidazodiazepine (5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazole[1,5-α][1,4]diazepin-3-yl) oxazole or KRM-II-81) is a potentiator of GABAA receptors (a GABAkine) undergoing preparation for clinical development. KRM-II-81 is active against many seizure and pain models in rodents, where it exhibits improved pharmacological properties over standard-of-care agents. Since salts can be utilized to create opportunities for increased solubility, enhanced absorption, and distribution, as well as for efficient methods of bulk synthesis, a hydrochloride salt of KRM-II-81 was prepared. KRM-II-81·HCl was produced from the free base with anhydrous hydrochloric acid. The formation of the monohydrochloride salt was confirmed by X-ray crystallography, as well as 1H NMR and 13C NMR analyses. High water solubility and a lower partition coefficient (octanol/water) were exhibited by KRM-II-81·HCl as compared to the free base. Oral administration of either KRM-II-81·HCl or the free base resulted in high concentrations in the brain and plasma of rats. Oral dosing in mice significantly increased the latency to both clonic and tonic convulsions and decreased pentylenetetrazol-induced lethality. The increased water solubility of the HCl salt enables intravenous dosing and the potential for higher concentration formulations compared with the free base without impacting anticonvulsant potency. Thus, KRM-II-81·HCl adds an important new compound to facilitate the development of these imidazodiazepines for clinical evaluation.
PB  - ACS Publications
T2  - ACS Omega
T1  - Hydrochloride Salt of the GABAkine KRM-II-81
VL  - 7
IS  - 31
SP  - 27550
EP  - 27559
DO  - 10.1021/acsomega.2c03029
ER  - 

@article{
author = "Mian, Md Yeunus and Divović, Branka and Sharmin, Dishary and Pandey, Kamal P. and Golani, Lalit K. and Tiruveedhula, V. V. N. Phani Babu and Cerne, Rok and Smith, Jodi L. and Ping, Xingjie and Jin, Xiaoming and Imler, Gregory H. and Deschamps, Jeffrey R. and Lippa, Arnold and Cook, James M. and Savić, Miroslav and Rowlett, James and Witkin, Jeffrey M.",
year = "2022",
abstract = "Imidazodiazepine (5-(8-ethynyl-6-(pyridin-2-yl)-4H-benzo[f]imidazole[1,5-α][1,4]diazepin-3-yl) oxazole or KRM-II-81) is a potentiator of GABAA receptors (a GABAkine) undergoing preparation for clinical development. KRM-II-81 is active against many seizure and pain models in rodents, where it exhibits improved pharmacological properties over standard-of-care agents. Since salts can be utilized to create opportunities for increased solubility, enhanced absorption, and distribution, as well as for efficient methods of bulk synthesis, a hydrochloride salt of KRM-II-81 was prepared. KRM-II-81·HCl was produced from the free base with anhydrous hydrochloric acid. The formation of the monohydrochloride salt was confirmed by X-ray crystallography, as well as 1H NMR and 13C NMR analyses. High water solubility and a lower partition coefficient (octanol/water) were exhibited by KRM-II-81·HCl as compared to the free base. Oral administration of either KRM-II-81·HCl or the free base resulted in high concentrations in the brain and plasma of rats. Oral dosing in mice significantly increased the latency to both clonic and tonic convulsions and decreased pentylenetetrazol-induced lethality. The increased water solubility of the HCl salt enables intravenous dosing and the potential for higher concentration formulations compared with the free base without impacting anticonvulsant potency. Thus, KRM-II-81·HCl adds an important new compound to facilitate the development of these imidazodiazepines for clinical evaluation.",
publisher = "ACS Publications",
journal = "ACS Omega",
title = "Hydrochloride Salt of the GABAkine KRM-II-81",
volume = "7",
number = "31",
pages = "27550-27559",
doi = "10.1021/acsomega.2c03029"
}

Mian, M. Y., Divović, B., Sharmin, D., Pandey, K. P., Golani, L. K., Tiruveedhula, V. V. N. P. B., Cerne, R., Smith, J. L., Ping, X., Jin, X., Imler, G. H., Deschamps, J. R., Lippa, A., Cook, J. M., Savić, M., Rowlett, J.,& Witkin, J. M.. (2022). Hydrochloride Salt of the GABAkine KRM-II-81. in ACS Omega
ACS Publications., 7(31), 27550-27559.
https://doi.org/10.1021/acsomega.2c03029

Mian MY, Divović B, Sharmin D, Pandey KP, Golani LK, Tiruveedhula VVNPB, Cerne R, Smith JL, Ping X, Jin X, Imler GH, Deschamps JR, Lippa A, Cook JM, Savić M, Rowlett J, Witkin JM. Hydrochloride Salt of the GABAkine KRM-II-81. in ACS Omega. 2022;7(31):27550-27559.
doi:10.1021/acsomega.2c03029 .

Mian, Md Yeunus, Divović, Branka, Sharmin, Dishary, Pandey, Kamal P., Golani, Lalit K., Tiruveedhula, V. V. N. Phani Babu, Cerne, Rok, Smith, Jodi L., Ping, Xingjie, Jin, Xiaoming, Imler, Gregory H., Deschamps, Jeffrey R., Lippa, Arnold, Cook, James M., Savić, Miroslav, Rowlett, James, Witkin, Jeffrey M., "Hydrochloride Salt of the GABAkine KRM-II-81" in ACS Omega, 7, no. 31 (2022):27550-27559,
https://doi.org/10.1021/acsomega.2c03029 . .

TY  - JOUR
AU  - Radojević, Branislava
AU  - Dragašević-Mišković, Nataša T.
AU  - Milovanović, Andona
AU  - Svetel, Marina
AU  - Petrović, Igor
AU  - Pešić, Maja
AU  - Tomić, Aleksandra
AU  - Stanisavljević, Dejana
AU  - Savić, Miroslav
AU  - Kostić, Vladimir S.
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4329
AB  - Objectives: Adherence to medication is an important factor that can influence Parkinson's disease (PD) control. We aimed to explore patients' adherence to antiparkinsonian medication and determine factors that might affect adherence to medications among PD patients. Methods: A cross-sectional, exploratory survey of PD patients treated with at least one antiparkinsonian drug and with a total score of MoCA (Montreal Cognitive Assessment) ≥26 was conducted. The final sample included 112 PD patients. A patient's adherence was assessed through ARMS (Adherence to Refills and Medications Scale). ARMS scores higher than 12 were assumed lower adherence. In addition, each patient underwent neurological examination, assessment of depression, anxiety, and evaluation of the presence of PD nonmotor symptoms. Results: The mean ARDS value in our cohort was 14.9 ± 2.5. Most PD patients (74.1%) reported lower adherence to their medication. Participants in the lower adherence group were younger at PD onset, had significantly higher UPDRS (Unified PD Rating Scale) scores, as well as UPDRS III and UPDRS IV subscores, HARS (Hamilton Anxiety Rating Scale), and NMSQuest (Non-Motor Symptoms Questionnaire for PD) scores compared to the fully adherent group (p=0.013, p=0.017, p=0.041, p=0.043, and p=0.023, respectively). Among nonmotor PD symptoms, the presence of cardiovascular, apathy/attention-deficit/memory disorders, hallucinations/delusions, and problems regarding changes in weight, diplopia, or sweating were associated with lower adherence. Multivariate regression analysis revealed depression as the strongest independent predictor of lower adherence. Conclusion: Depressed PD patients compared to PD patients without clinical depression had a three times higher risk for lower adherence to pharmacotherapy. Recognition and adequate treatment of depression might result in improved adherence.
PB  - Hindawi
T2  - International journal of clinical practice
T1  - Adherence to Medication among Parkinson's Disease Patients Using the Adherence to Refills and Medications Scale
VL  - 2022
DO  - 10.1155/2022/6741280
ER  - 

@article{
author = "Radojević, Branislava and Dragašević-Mišković, Nataša T. and Milovanović, Andona and Svetel, Marina and Petrović, Igor and Pešić, Maja and Tomić, Aleksandra and Stanisavljević, Dejana and Savić, Miroslav and Kostić, Vladimir S.",
year = "2022",
abstract = "Objectives: Adherence to medication is an important factor that can influence Parkinson's disease (PD) control. We aimed to explore patients' adherence to antiparkinsonian medication and determine factors that might affect adherence to medications among PD patients. Methods: A cross-sectional, exploratory survey of PD patients treated with at least one antiparkinsonian drug and with a total score of MoCA (Montreal Cognitive Assessment) ≥26 was conducted. The final sample included 112 PD patients. A patient's adherence was assessed through ARMS (Adherence to Refills and Medications Scale). ARMS scores higher than 12 were assumed lower adherence. In addition, each patient underwent neurological examination, assessment of depression, anxiety, and evaluation of the presence of PD nonmotor symptoms. Results: The mean ARDS value in our cohort was 14.9 ± 2.5. Most PD patients (74.1%) reported lower adherence to their medication. Participants in the lower adherence group were younger at PD onset, had significantly higher UPDRS (Unified PD Rating Scale) scores, as well as UPDRS III and UPDRS IV subscores, HARS (Hamilton Anxiety Rating Scale), and NMSQuest (Non-Motor Symptoms Questionnaire for PD) scores compared to the fully adherent group (p=0.013, p=0.017, p=0.041, p=0.043, and p=0.023, respectively). Among nonmotor PD symptoms, the presence of cardiovascular, apathy/attention-deficit/memory disorders, hallucinations/delusions, and problems regarding changes in weight, diplopia, or sweating were associated with lower adherence. Multivariate regression analysis revealed depression as the strongest independent predictor of lower adherence. Conclusion: Depressed PD patients compared to PD patients without clinical depression had a three times higher risk for lower adherence to pharmacotherapy. Recognition and adequate treatment of depression might result in improved adherence.",
publisher = "Hindawi",
journal = "International journal of clinical practice",
title = "Adherence to Medication among Parkinson's Disease Patients Using the Adherence to Refills and Medications Scale",
volume = "2022",
doi = "10.1155/2022/6741280"
}

Radojević, B., Dragašević-Mišković, N. T., Milovanović, A., Svetel, M., Petrović, I., Pešić, M., Tomić, A., Stanisavljević, D., Savić, M.,& Kostić, V. S.. (2022). Adherence to Medication among Parkinson's Disease Patients Using the Adherence to Refills and Medications Scale. in International journal of clinical practice
Hindawi., 2022.
https://doi.org/10.1155/2022/6741280

Radojević B, Dragašević-Mišković NT, Milovanović A, Svetel M, Petrović I, Pešić M, Tomić A, Stanisavljević D, Savić M, Kostić VS. Adherence to Medication among Parkinson's Disease Patients Using the Adherence to Refills and Medications Scale. in International journal of clinical practice. 2022;2022.
doi:10.1155/2022/6741280 .

Radojević, Branislava, Dragašević-Mišković, Nataša T., Milovanović, Andona, Svetel, Marina, Petrović, Igor, Pešić, Maja, Tomić, Aleksandra, Stanisavljević, Dejana, Savić, Miroslav, Kostić, Vladimir S., "Adherence to Medication among Parkinson's Disease Patients Using the Adherence to Refills and Medications Scale" in International journal of clinical practice, 2022 (2022),
https://doi.org/10.1155/2022/6741280 . .

TY  - CONF
AU  - Mitrović, Jelena
AU  - Bjelošević, Maja
AU  - Knutson, Daniel E.
AU  - Kremenović, Aleksandar
AU  - Lunter, Dominique
AU  - Ahlin Grabnar, Pegi
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4270
AB  - 1.	INTRODUCTION 
Nanocrystal dispersions are considered as the universal formulation strategy for brick dust substances. However, the stability of these systems to aggregation represents a big issue. To overcome this, nanocrystal dispersions are usually solidified by freeze-drying (lyophilization). During this process the risk of aggregation is considered to be high, due to ice formation and/or water loss. To prevent the aggregation, For the particle size preservation, therefore, it is necessary to add cryoprotectants/lyoprotectants, among which sugars are most commonly used. To ensure good structure of the cake, bulking agents are often included in formulations, as well [1,2], although in nanocrystalline dispersions the combination of cryoprotectants and bulking agents is not frequent nor much investigated.
Nanocrystals of DK-I-56-1 (7‑methoxy‑2-(4‑methoxy‑d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one), patent protected pyrazoloquinolinone ligand, have been developed recently, and characterized in terms of physicochemical properties and pharmacokinetics after intraperitoneal administration in mice. These formulations were stable for three weeks [3]. Our aim in this study was to improve the stability by freeze-drying, and investigate the influence of different concentrations and physical form of cryoprotectants (sucrose, trehalose) and bulking agent (mannitol) as well as different primary drying conditions on the aggregation prevention.

2. MATERIALS AND METHODS
2.1. Materials 
DK-I-56-1 was synthesized at the Department of Chemistry and Biochemistry, University of Wisconsin—Milwaukee, USA. The following other materials were used: polysorbate 80, poloxamer 407, sucrose, mannitol (Sigma-Aldrich Laborchemikalien GmbH, Germany) and trehalose (Carl Roth GmbH, Germany). 
2.2. Lyophilization 
Nanocrystal dispersions stabilized by polysorbate 80 and poloxamer 407 were prepared by wet ball milling [3]. After addition of mannitol (M), sucrose (S), or trehalose (T) alone or in combination samples were freeze- dried. Two processes were applied: (1) freezing at -80 °C (3 h), primary drying at -10 °C, 0.340 mbar, secondary drying at 25 °C (24 h) or (2) freezing at -50 °C (3 h), primary drying at -45 °C, 0.2 mbar (21 h), secondary drying at 20 °C (30 h). Samples were stored in crimped vials at 25 °C (lyophilization 1) or 2-8 ºC (lyophilization 2) for three months.
2.3. Physicochemical characterization
Particle size (z-ave) was measured by Zetasizer Nano ZS (Malvern Instruments, UK) and Mastersizer (Malvern Mastersizer 2000 Malvern, UK). Redispersibility index (RDI) was calculated as z-ave (before)/z-ave (after) and expressed in percentages. Physical state of samples was determined by differential scanning calorimetry (DSC1; Mettler Toledo, Switzerland),powder X-ray diffraction (Rigaku Smartlab X-ray Diffractometer) and polarized light microscopy (PLM) (Carl Zeiss ApoTome Imager Z1 microscope Zeiss, Germany). 
3. RESULTS AND DISCUSSION
Right after preparation, nanocrystal dispersions were with submicron particle size around 160 nm, and PDI below 0.2, suggesting narrow size distribution. In the cryoprotectant screening phase, sucrose and/or mannitol were added in different concentrations. It was shown that 10% of the total stabilizer concentration was needed for the particle size preservation: the achieved RDI was above 95%, while cakes with sucrose alone or in combination with mannitol in ratio 1:1 or 3:2 were also with satisfied appearance (Figure 1).  
Lyophilization was conducted above or below the glass transition temperature of the maximally freeze-concentrated solution (Tg’) (around -39 ºC). When primary drying was performed at -10 °C, no aggregation was noticed right after lyophilization, but particle size increased significantly, lowering down the RDI to < 50%, after one month storage at 25 °C. This was confirmed by laser diffraction. In lyophilization 2, with primary drying at temperature below Tg’, trehalose was also used in the same concentration as sucrose and in combination with mannitol. Interestingly, in this process parameters setup, sucrose or trehalose alone did not prevent aggregation during freeze-drying. Particle size remained almost unchanged in formulation S+M 3+2 (RDI 95%) or slightly higher in T+M 3+2 (RDI 90%), after three months storage, suggesting it was most probably the optimal combination for the stabilization. 
Physical state analysis revealed that sucrose and mannitol in samples lyophilized by process 1 were in crystalline state, as well as sucrose when used alone in lyophilization 2. Trehalose, on the other hand was amorphous in all samples containing it. Amorphous state of lyoprotectants allows maximal hydrogen bonding due to higher molecule flexibility and availability of hydroxyl groups [3]. Surprisingly, mannitol as a substance with high crystallization tendency was with low crystallinity in lyophilizates. These observations were confirmed by PLM. It is possible that it formed interactions with sucrose or nanocrystal stabilizers [4]. 
4. CONCLUSION
Results from this study demonstrated freeze- drying as an important technique for the improvement of nanocrystals stability. However, the selection of cryoprotectant and bulking agent ratio beside process parameters (primary drying at -45 ºC) was crucial to get freeze-dried samples with good stability. Sucrose or trehalose in combination with mannitol (ratio 3+2) in total concentration 10% successfully hindered aggregation, thus prolonging the stability to 3 months at 2-8 ºC.
5. REFERENCES
1.	Van Eerdenbrugh, B., et al. Top-down production of drug nanocrystals: nanosuspension stabilization, miniaturization and transformation into solid products. International journal of pharmaceutics, 2008. 364(1): 64-75.
2.	Trenkenschuh, E., and Friess, W. Freeze-drying of nanoparticles: How to overcome colloidal instability by formulation and process optimization. European Journal of Pharmaceutics and Biopharmaceutics, 2021.165: 345-360.
3.	Mitrović, J.R., et al. Overcoming the low oral bioavailability of deuterated pyrazoloquinolinone ligand DK-I-60-3 by nanonization: A knowledge-based approach. Pharmaceutics, 2021. 13(8): 1188.
4.	Kumar, S., et al. Sugars as bulking agents to prevent nano-crystal aggregation during spray or freeze-drying. International journal of pharmaceutics, 2014. 471(1-2): 303-311.
ACKNOWLEDGMENT
This research was supported by the Science Fund of the Republic of Serbia, grant No. 7749108, project Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms-NanoCellEmoCog.
C3  - 9th BBBB International conference on pharmaceutical sciences; 15th - 17th September, Ljubljana, Slovenia
T1  - Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): process parameters and cryoprotectant selection through stability study
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4270
ER  - 

@conference{
author = "Mitrović, Jelena and Bjelošević, Maja and Knutson, Daniel E. and Kremenović, Aleksandar and Lunter, Dominique and Ahlin Grabnar, Pegi and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2022",
abstract = "1.	INTRODUCTION 
Nanocrystal dispersions are considered as the universal formulation strategy for brick dust substances. However, the stability of these systems to aggregation represents a big issue. To overcome this, nanocrystal dispersions are usually solidified by freeze-drying (lyophilization). During this process the risk of aggregation is considered to be high, due to ice formation and/or water loss. To prevent the aggregation, For the particle size preservation, therefore, it is necessary to add cryoprotectants/lyoprotectants, among which sugars are most commonly used. To ensure good structure of the cake, bulking agents are often included in formulations, as well [1,2], although in nanocrystalline dispersions the combination of cryoprotectants and bulking agents is not frequent nor much investigated.
Nanocrystals of DK-I-56-1 (7‑methoxy‑2-(4‑methoxy‑d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one), patent protected pyrazoloquinolinone ligand, have been developed recently, and characterized in terms of physicochemical properties and pharmacokinetics after intraperitoneal administration in mice. These formulations were stable for three weeks [3]. Our aim in this study was to improve the stability by freeze-drying, and investigate the influence of different concentrations and physical form of cryoprotectants (sucrose, trehalose) and bulking agent (mannitol) as well as different primary drying conditions on the aggregation prevention.

2. MATERIALS AND METHODS
2.1. Materials 
DK-I-56-1 was synthesized at the Department of Chemistry and Biochemistry, University of Wisconsin—Milwaukee, USA. The following other materials were used: polysorbate 80, poloxamer 407, sucrose, mannitol (Sigma-Aldrich Laborchemikalien GmbH, Germany) and trehalose (Carl Roth GmbH, Germany). 
2.2. Lyophilization 
Nanocrystal dispersions stabilized by polysorbate 80 and poloxamer 407 were prepared by wet ball milling [3]. After addition of mannitol (M), sucrose (S), or trehalose (T) alone or in combination samples were freeze- dried. Two processes were applied: (1) freezing at -80 °C (3 h), primary drying at -10 °C, 0.340 mbar, secondary drying at 25 °C (24 h) or (2) freezing at -50 °C (3 h), primary drying at -45 °C, 0.2 mbar (21 h), secondary drying at 20 °C (30 h). Samples were stored in crimped vials at 25 °C (lyophilization 1) or 2-8 ºC (lyophilization 2) for three months.
2.3. Physicochemical characterization
Particle size (z-ave) was measured by Zetasizer Nano ZS (Malvern Instruments, UK) and Mastersizer (Malvern Mastersizer 2000 Malvern, UK). Redispersibility index (RDI) was calculated as z-ave (before)/z-ave (after) and expressed in percentages. Physical state of samples was determined by differential scanning calorimetry (DSC1; Mettler Toledo, Switzerland),powder X-ray diffraction (Rigaku Smartlab X-ray Diffractometer) and polarized light microscopy (PLM) (Carl Zeiss ApoTome Imager Z1 microscope Zeiss, Germany). 
3. RESULTS AND DISCUSSION
Right after preparation, nanocrystal dispersions were with submicron particle size around 160 nm, and PDI below 0.2, suggesting narrow size distribution. In the cryoprotectant screening phase, sucrose and/or mannitol were added in different concentrations. It was shown that 10% of the total stabilizer concentration was needed for the particle size preservation: the achieved RDI was above 95%, while cakes with sucrose alone or in combination with mannitol in ratio 1:1 or 3:2 were also with satisfied appearance (Figure 1).  
Lyophilization was conducted above or below the glass transition temperature of the maximally freeze-concentrated solution (Tg’) (around -39 ºC). When primary drying was performed at -10 °C, no aggregation was noticed right after lyophilization, but particle size increased significantly, lowering down the RDI to < 50%, after one month storage at 25 °C. This was confirmed by laser diffraction. In lyophilization 2, with primary drying at temperature below Tg’, trehalose was also used in the same concentration as sucrose and in combination with mannitol. Interestingly, in this process parameters setup, sucrose or trehalose alone did not prevent aggregation during freeze-drying. Particle size remained almost unchanged in formulation S+M 3+2 (RDI 95%) or slightly higher in T+M 3+2 (RDI 90%), after three months storage, suggesting it was most probably the optimal combination for the stabilization. 
Physical state analysis revealed that sucrose and mannitol in samples lyophilized by process 1 were in crystalline state, as well as sucrose when used alone in lyophilization 2. Trehalose, on the other hand was amorphous in all samples containing it. Amorphous state of lyoprotectants allows maximal hydrogen bonding due to higher molecule flexibility and availability of hydroxyl groups [3]. Surprisingly, mannitol as a substance with high crystallization tendency was with low crystallinity in lyophilizates. These observations were confirmed by PLM. It is possible that it formed interactions with sucrose or nanocrystal stabilizers [4]. 
4. CONCLUSION
Results from this study demonstrated freeze- drying as an important technique for the improvement of nanocrystals stability. However, the selection of cryoprotectant and bulking agent ratio beside process parameters (primary drying at -45 ºC) was crucial to get freeze-dried samples with good stability. Sucrose or trehalose in combination with mannitol (ratio 3+2) in total concentration 10% successfully hindered aggregation, thus prolonging the stability to 3 months at 2-8 ºC.
5. REFERENCES
1.	Van Eerdenbrugh, B., et al. Top-down production of drug nanocrystals: nanosuspension stabilization, miniaturization and transformation into solid products. International journal of pharmaceutics, 2008. 364(1): 64-75.
2.	Trenkenschuh, E., and Friess, W. Freeze-drying of nanoparticles: How to overcome colloidal instability by formulation and process optimization. European Journal of Pharmaceutics and Biopharmaceutics, 2021.165: 345-360.
3.	Mitrović, J.R., et al. Overcoming the low oral bioavailability of deuterated pyrazoloquinolinone ligand DK-I-60-3 by nanonization: A knowledge-based approach. Pharmaceutics, 2021. 13(8): 1188.
4.	Kumar, S., et al. Sugars as bulking agents to prevent nano-crystal aggregation during spray or freeze-drying. International journal of pharmaceutics, 2014. 471(1-2): 303-311.
ACKNOWLEDGMENT
This research was supported by the Science Fund of the Republic of Serbia, grant No. 7749108, project Neuroimmune aspects of mood, anxiety and cognitive effects of leads/drug candidates acting at GABAA and/or sigma-2 receptors: In vitro/in vivo delineation by nano- and hiPSC-based platforms-NanoCellEmoCog.",
journal = "9th BBBB International conference on pharmaceutical sciences; 15th - 17th September, Ljubljana, Slovenia",
title = "Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): process parameters and cryoprotectant selection through stability study",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4270"
}

Mitrović, J., Bjelošević, M., Knutson, D. E., Kremenović, A., Lunter, D., Ahlin Grabnar, P., Cook, J. M., Savić, M.,& Savić, S.. (2022). Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): process parameters and cryoprotectant selection through stability study. in 9th BBBB International conference on pharmaceutical sciences; 15th - 17th September, Ljubljana, Slovenia.
https://hdl.handle.net/21.15107/rcub_farfar_4270

Mitrović J, Bjelošević M, Knutson DE, Kremenović A, Lunter D, Ahlin Grabnar P, Cook JM, Savić M, Savić S. Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): process parameters and cryoprotectant selection through stability study. in 9th BBBB International conference on pharmaceutical sciences; 15th - 17th September, Ljubljana, Slovenia. 2022;.
https://hdl.handle.net/21.15107/rcub_farfar_4270 .

Mitrović, Jelena, Bjelošević, Maja, Knutson, Daniel E., Kremenović, Aleksandar, Lunter, Dominique, Ahlin Grabnar, Pegi, Cook, James M., Savić, Miroslav, Savić, Snežana, "Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): process parameters and cryoprotectant selection through stability study" in 9th BBBB International conference on pharmaceutical sciences; 15th - 17th September, Ljubljana, Slovenia (2022),
https://hdl.handle.net/21.15107/rcub_farfar_4270 .

TY  - JOUR
AU  - Aranđelović, Jovana
AU  - Santrač, Anja
AU  - Batinić, Bojan
AU  - Todorović, Lidija
AU  - Stevanović, Vladimir
AU  - Tiruveedhula, Veera Venkata Naga Phani Babu
AU  - Sharmin, Dishary
AU  - Rashid, Farjana
AU  - Stanojević, Boban
AU  - Cook, James
AU  - Savić, Miroslav
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4203
AB  - Aims: GABAergic modulation involved in cognitive processing appears to be substan- tially changed in Alzheimer's disease (AD). In a widely used 5xFAD model of AD, we aimed to assess if negative and positive allosteric modulators of α5 GABA A receptors (NAM and PAM, respectively) would affect social interaction, social, object and spatial memory, and neuroinflammation. Methods: After 10-day treatment with PAM, NAM, or solvent, 6-month-old trans- genic and non-transgenic 5xFAD mice underwent testing in a behavioral battery. Gene expressions of IL-1β, IL-6, TNF-α, GFAP, and IBA-1 were determined in hippocampus and prefrontal cortex by qPCR. Results: PAM treatment impaired spatial learning in transgenic females compared to solvent-treated transgenic females, and social recognition in transgenic and non- transgenic males. NAM treatment declined social interaction in transgenic and non- transgenic males, while had beneficial effect on cognitive flexibility in non-transgenic males compared to solvent-treated non-transgenic males. Transgenic animals have not fully displayed cognitive symptoms, but neuroinflammation was confirmed. NAM reduced proinflammatory gene expressions in transgenic females and astrogliosis in transgenic males compared to pathological controls. Conclusion: PAM and NAM failed to exert favorable behavioral effects in transgenic animals. Suppression of neuroinflammation obtained with NAM calls for more studies with GABAergic ligands in amyloid beta- and/or tau-dependent models with promi- nent neuroinflammation.
PB  - John Wiley and Sons Inc
T2  - CNS Neuroscience & Therapeutics
T1  - Effects of α5 GABAA receptor modulation on social interaction, memory, and neuroinflammation in a mouse model of Alzheimer's disease
VL  - 28
IS  - 11
SP  - 1767
EP  - 1778
DO  - 10.1111/cns.13914
ER  - 

@article{
author = "Aranđelović, Jovana and Santrač, Anja and Batinić, Bojan and Todorović, Lidija and Stevanović, Vladimir and Tiruveedhula, Veera Venkata Naga Phani Babu and Sharmin, Dishary and Rashid, Farjana and Stanojević, Boban and Cook, James and Savić, Miroslav",
year = "2022",
abstract = "Aims: GABAergic modulation involved in cognitive processing appears to be substan- tially changed in Alzheimer's disease (AD). In a widely used 5xFAD model of AD, we aimed to assess if negative and positive allosteric modulators of α5 GABA A receptors (NAM and PAM, respectively) would affect social interaction, social, object and spatial memory, and neuroinflammation. Methods: After 10-day treatment with PAM, NAM, or solvent, 6-month-old trans- genic and non-transgenic 5xFAD mice underwent testing in a behavioral battery. Gene expressions of IL-1β, IL-6, TNF-α, GFAP, and IBA-1 were determined in hippocampus and prefrontal cortex by qPCR. Results: PAM treatment impaired spatial learning in transgenic females compared to solvent-treated transgenic females, and social recognition in transgenic and non- transgenic males. NAM treatment declined social interaction in transgenic and non- transgenic males, while had beneficial effect on cognitive flexibility in non-transgenic males compared to solvent-treated non-transgenic males. Transgenic animals have not fully displayed cognitive symptoms, but neuroinflammation was confirmed. NAM reduced proinflammatory gene expressions in transgenic females and astrogliosis in transgenic males compared to pathological controls. Conclusion: PAM and NAM failed to exert favorable behavioral effects in transgenic animals. Suppression of neuroinflammation obtained with NAM calls for more studies with GABAergic ligands in amyloid beta- and/or tau-dependent models with promi- nent neuroinflammation.",
publisher = "John Wiley and Sons Inc",
journal = "CNS Neuroscience & Therapeutics",
title = "Effects of α5 GABAA receptor modulation on social interaction, memory, and neuroinflammation in a mouse model of Alzheimer's disease",
volume = "28",
number = "11",
pages = "1767-1778",
doi = "10.1111/cns.13914"
}

Aranđelović, J., Santrač, A., Batinić, B., Todorović, L., Stevanović, V., Tiruveedhula, V. V. N. P. B., Sharmin, D., Rashid, F., Stanojević, B., Cook, J.,& Savić, M.. (2022). Effects of α5 GABAA receptor modulation on social interaction, memory, and neuroinflammation in a mouse model of Alzheimer's disease. in CNS Neuroscience & Therapeutics
John Wiley and Sons Inc., 28(11), 1767-1778.
https://doi.org/10.1111/cns.13914

Aranđelović J, Santrač A, Batinić B, Todorović L, Stevanović V, Tiruveedhula VVNPB, Sharmin D, Rashid F, Stanojević B, Cook J, Savić M. Effects of α5 GABAA receptor modulation on social interaction, memory, and neuroinflammation in a mouse model of Alzheimer's disease. in CNS Neuroscience & Therapeutics. 2022;28(11):1767-1778.
doi:10.1111/cns.13914 .

Aranđelović, Jovana, Santrač, Anja, Batinić, Bojan, Todorović, Lidija, Stevanović, Vladimir, Tiruveedhula, Veera Venkata Naga Phani Babu, Sharmin, Dishary, Rashid, Farjana, Stanojević, Boban, Cook, James, Savić, Miroslav, "Effects of α5 GABAA receptor modulation on social interaction, memory, and neuroinflammation in a mouse model of Alzheimer's disease" in CNS Neuroscience & Therapeutics, 28, no. 11 (2022):1767-1778,
https://doi.org/10.1111/cns.13914 . .

TY  - JOUR
AU  - Golani, Lalit
AU  - Divović, Branka
AU  - Sharmin, Dishary
AU  - Pandey, Kamal
AU  - Mian, Md Yeunus
AU  - Cerne, Rok
AU  - Zahn, Nicolas
AU  - Meyer, Michelle
AU  - Tiruveedhula, Veera
AU  - Smith, Jodi
AU  - Ping, Xingjie
AU  - Jin, Xiaoming
AU  - Lippa, Arnold
AU  - Schkeryantz, Jeffrey
AU  - Arnold, Leggy
AU  - Cook, James
AU  - Savić, Miroslav
AU  - Witkin, Jeffrey
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4108
AB  - The imidazodiazepine, (5‐(8‐ethynyl‐6‐(pyridin‐2‐yl)‐4H‐benzo [f]imidazole[1,5‐α] [1,4]diazepin‐3‐yl) oxazole or KRM‐II‐81) is a new α2/3‐selective GABAkine (gamma aminobutyric acid A receptor potentiator) with anticonvulsant, anxiolytic, and antinociceptive activity in preclinical models. Reducing metabolism was utilized as a means of potentially extending the half‐life of KRM‐II‐81. In vitro and in vivo studies were conducted to evaluate metabolic liabilities. Incubation of KRM‐II‐81 in hepatocytes revealed sites of potential metabolism on the oxazole and the diazepine rings. These sites were targeted in the design of a deuterated analog (D5‐KRM‐II‐ 81) that could be evaluated as a potentially longer‐acting analog. In contrast to computer predictions, peak plasma concentrations of D5‐KRM‐II‐81 in rats were not significantly greater than those produced by KRM‐II‐81 after oral administra- tion. Furthermore, brain disposition of KRM‐II‐81 was higher than that of D5‐KRM‐ II‐81. The half‐life of the two compounds in either plasma or brain did not statis- tically differ from one another but the tmax for D5‐KRM‐II‐81 occurred slightly earlier than for KRM‐II‐81. Non‐metabolic considerations might be relevant to the lack of increases in exposure by D5‐KRM‐II‐81. Alternative sites of metabolism on KRM‐II‐81, not targeted by the current deuteration process, are also possible. Despite its lack of augmented exposure, D5‐KRM‐II‐81, like KRM‐II‐81, significantly prevented seizures induced by pentylenetetrazol when given orally. The present findings introduce a new orally active anticonvulsant GABAkine, D5‐KRM‐II‐81.
PB  - John Wiley and Sons Ltd
T2  - Biopharmaceutics and Drug Disposition
T1  - Metabolism, pharmacokinetics, and anticonvulsant activity of a deuterated analog of the α2/3-selective GABAkine KRM-II-81
VL  - 43
IS  - 2
SP  - 66
EP  - 75
DO  - 10.1002/bdd.2313
ER  - 

@article{
author = "Golani, Lalit and Divović, Branka and Sharmin, Dishary and Pandey, Kamal and Mian, Md Yeunus and Cerne, Rok and Zahn, Nicolas and Meyer, Michelle and Tiruveedhula, Veera and Smith, Jodi and Ping, Xingjie and Jin, Xiaoming and Lippa, Arnold and Schkeryantz, Jeffrey and Arnold, Leggy and Cook, James and Savić, Miroslav and Witkin, Jeffrey",
year = "2022",
abstract = "The imidazodiazepine, (5‐(8‐ethynyl‐6‐(pyridin‐2‐yl)‐4H‐benzo [f]imidazole[1,5‐α] [1,4]diazepin‐3‐yl) oxazole or KRM‐II‐81) is a new α2/3‐selective GABAkine (gamma aminobutyric acid A receptor potentiator) with anticonvulsant, anxiolytic, and antinociceptive activity in preclinical models. Reducing metabolism was utilized as a means of potentially extending the half‐life of KRM‐II‐81. In vitro and in vivo studies were conducted to evaluate metabolic liabilities. Incubation of KRM‐II‐81 in hepatocytes revealed sites of potential metabolism on the oxazole and the diazepine rings. These sites were targeted in the design of a deuterated analog (D5‐KRM‐II‐ 81) that could be evaluated as a potentially longer‐acting analog. In contrast to computer predictions, peak plasma concentrations of D5‐KRM‐II‐81 in rats were not significantly greater than those produced by KRM‐II‐81 after oral administra- tion. Furthermore, brain disposition of KRM‐II‐81 was higher than that of D5‐KRM‐ II‐81. The half‐life of the two compounds in either plasma or brain did not statis- tically differ from one another but the tmax for D5‐KRM‐II‐81 occurred slightly earlier than for KRM‐II‐81. Non‐metabolic considerations might be relevant to the lack of increases in exposure by D5‐KRM‐II‐81. Alternative sites of metabolism on KRM‐II‐81, not targeted by the current deuteration process, are also possible. Despite its lack of augmented exposure, D5‐KRM‐II‐81, like KRM‐II‐81, significantly prevented seizures induced by pentylenetetrazol when given orally. The present findings introduce a new orally active anticonvulsant GABAkine, D5‐KRM‐II‐81.",
publisher = "John Wiley and Sons Ltd",
journal = "Biopharmaceutics and Drug Disposition",
title = "Metabolism, pharmacokinetics, and anticonvulsant activity of a deuterated analog of the α2/3-selective GABAkine KRM-II-81",
volume = "43",
number = "2",
pages = "66-75",
doi = "10.1002/bdd.2313"
}

Golani, L., Divović, B., Sharmin, D., Pandey, K., Mian, M. Y., Cerne, R., Zahn, N., Meyer, M., Tiruveedhula, V., Smith, J., Ping, X., Jin, X., Lippa, A., Schkeryantz, J., Arnold, L., Cook, J., Savić, M.,& Witkin, J.. (2022). Metabolism, pharmacokinetics, and anticonvulsant activity of a deuterated analog of the α2/3-selective GABAkine KRM-II-81. in Biopharmaceutics and Drug Disposition
John Wiley and Sons Ltd., 43(2), 66-75.
https://doi.org/10.1002/bdd.2313

Golani L, Divović B, Sharmin D, Pandey K, Mian MY, Cerne R, Zahn N, Meyer M, Tiruveedhula V, Smith J, Ping X, Jin X, Lippa A, Schkeryantz J, Arnold L, Cook J, Savić M, Witkin J. Metabolism, pharmacokinetics, and anticonvulsant activity of a deuterated analog of the α2/3-selective GABAkine KRM-II-81. in Biopharmaceutics and Drug Disposition. 2022;43(2):66-75.
doi:10.1002/bdd.2313 .

Golani, Lalit, Divović, Branka, Sharmin, Dishary, Pandey, Kamal, Mian, Md Yeunus, Cerne, Rok, Zahn, Nicolas, Meyer, Michelle, Tiruveedhula, Veera, Smith, Jodi, Ping, Xingjie, Jin, Xiaoming, Lippa, Arnold, Schkeryantz, Jeffrey, Arnold, Leggy, Cook, James, Savić, Miroslav, Witkin, Jeffrey, "Metabolism, pharmacokinetics, and anticonvulsant activity of a deuterated analog of the α2/3-selective GABAkine KRM-II-81" in Biopharmaceutics and Drug Disposition, 43, no. 2 (2022):66-75,
https://doi.org/10.1002/bdd.2313 . .

TY  - JOUR
AU  - Sieghart, Werner
AU  - Chiou, Lih-Chu
AU  - Ernst, Margot
AU  - Fabjan, Jure
AU  - Savić, Miroslav
AU  - Lee, Ming Tatt
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4107
AB  - GABAA receptors containing the α6 subunit are highly expressed in cerebellar granule cells and less abundantly in many other neuronal and peripheral tissues. Here, we for the first time summarize their importance for the functions of the cerebellum and the nervous system. The cerebellum is not only involved in motor control but also in cognitive, emotional, and social behaviors. α6βγ2 GABAA receptors located at cerebellar Golgi cell/granule cell synapses enhance the precision of inputs required for cerebellar timing of motor activity and are thus involved in cognitive processing and adequate responses to our environment. Extrasynaptic α6βδ GABAA receptors regulate the amount of information entering the cerebellum by their tonic inhibition of granule cells, and their optimal functioning enhances input filtering or contrast. The complex roles of the cerebellum in multiple brain functions can be compromised by genetic or neurodevelopmental causes that lead to a hypofunction of cerebellar α6-containing GABAA receptors. Animal models mimicking neuropsychiatric phenotypes suggest that compounds selectively activating or positively modulating cerebellar α6-containing GABAA receptors can alleviate essential tremor and motor disturbances in Angelman and Down syndrome as well as impaired prepulse inhibition in neuropsychiatric disorders and reduce migraine and trigeminal-related pain via α6-containing GABAA receptors in trigeminal ganglia. Genetic studies in humans suggest an association of the human GABAA receptor α6 subunit gene with stress-associated disorders. Animal studies support this conclusion. Neuroimaging and post-mortem studies in humans further support an involvement of α6-containing GABAA receptors in various neuropsychiatric disorders, pointing to a broad therapeutic potential of drugs modulating α6-containing GABAA receptors.
PB  - American Society for Pharmacology and Experimental Therapy
T2  - Pharmacological Reviews
T1  - α6-Containing GABAA Receptors: Functional Roles and Therapeutic Potentials
VL  - 74
IS  - 1
SP  - 238
EP  - 270
DO  - 10.1124/PHARMREV.121.000293
ER  - 

@article{
author = "Sieghart, Werner and Chiou, Lih-Chu and Ernst, Margot and Fabjan, Jure and Savić, Miroslav and Lee, Ming Tatt",
year = "2022",
abstract = "GABAA receptors containing the α6 subunit are highly expressed in cerebellar granule cells and less abundantly in many other neuronal and peripheral tissues. Here, we for the first time summarize their importance for the functions of the cerebellum and the nervous system. The cerebellum is not only involved in motor control but also in cognitive, emotional, and social behaviors. α6βγ2 GABAA receptors located at cerebellar Golgi cell/granule cell synapses enhance the precision of inputs required for cerebellar timing of motor activity and are thus involved in cognitive processing and adequate responses to our environment. Extrasynaptic α6βδ GABAA receptors regulate the amount of information entering the cerebellum by their tonic inhibition of granule cells, and their optimal functioning enhances input filtering or contrast. The complex roles of the cerebellum in multiple brain functions can be compromised by genetic or neurodevelopmental causes that lead to a hypofunction of cerebellar α6-containing GABAA receptors. Animal models mimicking neuropsychiatric phenotypes suggest that compounds selectively activating or positively modulating cerebellar α6-containing GABAA receptors can alleviate essential tremor and motor disturbances in Angelman and Down syndrome as well as impaired prepulse inhibition in neuropsychiatric disorders and reduce migraine and trigeminal-related pain via α6-containing GABAA receptors in trigeminal ganglia. Genetic studies in humans suggest an association of the human GABAA receptor α6 subunit gene with stress-associated disorders. Animal studies support this conclusion. Neuroimaging and post-mortem studies in humans further support an involvement of α6-containing GABAA receptors in various neuropsychiatric disorders, pointing to a broad therapeutic potential of drugs modulating α6-containing GABAA receptors.",
publisher = "American Society for Pharmacology and Experimental Therapy",
journal = "Pharmacological Reviews",
title = "α6-Containing GABAA Receptors: Functional Roles and Therapeutic Potentials",
volume = "74",
number = "1",
pages = "238-270",
doi = "10.1124/PHARMREV.121.000293"
}

Sieghart, W., Chiou, L., Ernst, M., Fabjan, J., Savić, M.,& Lee, M. T.. (2022). α6-Containing GABAA Receptors: Functional Roles and Therapeutic Potentials. in Pharmacological Reviews
American Society for Pharmacology and Experimental Therapy., 74(1), 238-270.
https://doi.org/10.1124/PHARMREV.121.000293

Sieghart W, Chiou L, Ernst M, Fabjan J, Savić M, Lee MT. α6-Containing GABAA Receptors: Functional Roles and Therapeutic Potentials. in Pharmacological Reviews. 2022;74(1):238-270.
doi:10.1124/PHARMREV.121.000293 .

Sieghart, Werner, Chiou, Lih-Chu, Ernst, Margot, Fabjan, Jure, Savić, Miroslav, Lee, Ming Tatt, "α6-Containing GABAA Receptors: Functional Roles and Therapeutic Potentials" in Pharmacological Reviews, 74, no. 1 (2022):238-270,
https://doi.org/10.1124/PHARMREV.121.000293 . .

TY  - JOUR
AU  - Radojević, Branislava
AU  - Dragašević-Mišković, Nataša
AU  - Marjanović, Ana
AU  - Branković, Marija
AU  - Milovanović, Andona
AU  - Petrović, Igor
AU  - Svetel, Marina
AU  - Jančić, Ivan
AU  - Stanisavljević, Dejana
AU  - Milićević, Ognjen
AU  - Savić, Miroslav
AU  - Kostić, Vladimir
PY  - 2022
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4082
AB  - Background: Clinical-related risk factors to freezing of gait (FOG) in Parkinson’s disease (PD) have been iden- tified. Still, the influence of genetic variations on the FOG occurrence has been poorly studied thus far. Aim: We aimed to evaluate the association of six selected polymorphisms of DRD2, ANKK1, and COMT genes with the FOG occurrence and explore the influence of ANNK1/DRD2 haplotypes on the onset of FOG in the group of PD patients. Method: PD patients (n = 234), treated with levodopa for at least two years, were genotyped for the rs4680 in COMT, rs6277, rs1076560, and rs2283265 in DRD2, and rs1800497 and rs2734849 polymorphisms in ANKK1 genes. FOG was evaluated by posing a direct question. In addition, a comprehensive set of clinical scales was applied to all patients. Results: FOG occurred in 132 (56.4%) PD patients in our cohort. Freezers were younger at PD onset, had longer disease duration, used higher levodopa daily doses and dopaminergic agents, and had higher motor and non- motor scales scores than non-freezers. FOG was more frequent among AA rs4680 COMT carriers than AG and GG rs4680 COMT carriers. Independent predictors of FOG were: disease duration of more than ten years, levodopa daily dose higher than 500 mg/day, motor status, and COMT AA genotype. AGGAA and GGAAA haplotypes were revealed as protective and vulnerability factors for FOG occurrence. Conclusion: In addition to previously identified disease- and therapy-related risk factors, our results suggested a possible contribution of dopamine-related genes to the FOG occurrence.
PB  - Elsevier Ltd
T2  - Parkinsonism and Related Disorders
T1  - The correlation between genetic factors and freezing of gait in patients with Parkinson's disease
VL  - 98
SP  - 7
EP  - 12
DO  - 10.1016/j.parkreldis.2022.03.018
ER  - 

@article{
author = "Radojević, Branislava and Dragašević-Mišković, Nataša and Marjanović, Ana and Branković, Marija and Milovanović, Andona and Petrović, Igor and Svetel, Marina and Jančić, Ivan and Stanisavljević, Dejana and Milićević, Ognjen and Savić, Miroslav and Kostić, Vladimir",
year = "2022",
abstract = "Background: Clinical-related risk factors to freezing of gait (FOG) in Parkinson’s disease (PD) have been iden- tified. Still, the influence of genetic variations on the FOG occurrence has been poorly studied thus far. Aim: We aimed to evaluate the association of six selected polymorphisms of DRD2, ANKK1, and COMT genes with the FOG occurrence and explore the influence of ANNK1/DRD2 haplotypes on the onset of FOG in the group of PD patients. Method: PD patients (n = 234), treated with levodopa for at least two years, were genotyped for the rs4680 in COMT, rs6277, rs1076560, and rs2283265 in DRD2, and rs1800497 and rs2734849 polymorphisms in ANKK1 genes. FOG was evaluated by posing a direct question. In addition, a comprehensive set of clinical scales was applied to all patients. Results: FOG occurred in 132 (56.4%) PD patients in our cohort. Freezers were younger at PD onset, had longer disease duration, used higher levodopa daily doses and dopaminergic agents, and had higher motor and non- motor scales scores than non-freezers. FOG was more frequent among AA rs4680 COMT carriers than AG and GG rs4680 COMT carriers. Independent predictors of FOG were: disease duration of more than ten years, levodopa daily dose higher than 500 mg/day, motor status, and COMT AA genotype. AGGAA and GGAAA haplotypes were revealed as protective and vulnerability factors for FOG occurrence. Conclusion: In addition to previously identified disease- and therapy-related risk factors, our results suggested a possible contribution of dopamine-related genes to the FOG occurrence.",
publisher = "Elsevier Ltd",
journal = "Parkinsonism and Related Disorders",
title = "The correlation between genetic factors and freezing of gait in patients with Parkinson's disease",
volume = "98",
pages = "7-12",
doi = "10.1016/j.parkreldis.2022.03.018"
}

Radojević, B., Dragašević-Mišković, N., Marjanović, A., Branković, M., Milovanović, A., Petrović, I., Svetel, M., Jančić, I., Stanisavljević, D., Milićević, O., Savić, M.,& Kostić, V.. (2022). The correlation between genetic factors and freezing of gait in patients with Parkinson's disease. in Parkinsonism and Related Disorders
Elsevier Ltd., 98, 7-12.
https://doi.org/10.1016/j.parkreldis.2022.03.018

Radojević B, Dragašević-Mišković N, Marjanović A, Branković M, Milovanović A, Petrović I, Svetel M, Jančić I, Stanisavljević D, Milićević O, Savić M, Kostić V. The correlation between genetic factors and freezing of gait in patients with Parkinson's disease. in Parkinsonism and Related Disorders. 2022;98:7-12.
doi:10.1016/j.parkreldis.2022.03.018 .

Radojević, Branislava, Dragašević-Mišković, Nataša, Marjanović, Ana, Branković, Marija, Milovanović, Andona, Petrović, Igor, Svetel, Marina, Jančić, Ivan, Stanisavljević, Dejana, Milićević, Ognjen, Savić, Miroslav, Kostić, Vladimir, "The correlation between genetic factors and freezing of gait in patients with Parkinson's disease" in Parkinsonism and Related Disorders, 98 (2022):7-12,
https://doi.org/10.1016/j.parkreldis.2022.03.018 . .