TY  - JOUR
AU  - Živančević, Katarina
AU  - Baralić, Katarina
AU  - Vukelić, Dragana
AU  - Marić, Đurđica
AU  - Kotur-Stevuljević, Jelena
AU  - Ivanišević, Jasmina
AU  - Savić, Miroslav
AU  - Batinić, Bojan
AU  - Janković, Radmila
AU  - Buha-Đorđević, Aleksandra
AU  - Antonijević-Miljaković, Evica
AU  - Ćurčić, Marijana
AU  - Bulat, Zorica
AU  - Antonijević, Biljana
AU  - Đukić-Ćosić, Danijela
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5614
AB  - Metals exert detrimental effects on various systems within the body, including the nervous system. Nevertheless, the dose-response relationship concerning the administration of low doses of metal mixtures remains inadequately explored. The assessment of neurotoxic effects of lead, cadmium, mercury, and arsenic mixture (MIX) administered at low dose ranges, was conducted using an in vivo approach. A subacute study was conducted on a rat model consisting of a control and five treatment groups subjected to oral exposure with gradually increasing doses (from MIX 1 to MIX 5). The results indicated that behavioural patterns in an already developed nervous system displayed a reduced susceptibility to the metal mixture exposure with tendency of higher doses to alter short term memory. However, the vulnerability of the mature brain to even minimal amounts of the investigated metal mixture was evident, particularly in the context of oxidative stress. Moreover, the study highlights superoxide dismutase's sensitivity as an early-stage neurotoxicity marker, as indicated by dose-dependent induction of oxidative stress in the brain revealed through Benchmark analysis. The narrowest Benchmark Dose Interval (BMDI) for superoxide dismutase (SOD) activity (1e-06 - 3.18e-05 mg As/kg b.w./day) indicates that arsenic may dictate the alterations in SOD activity when co-exposed with the other examined metals. The predicted Benchmark doses for oxidative stress parameters were very low, supporting “no-threshold” concept. Histopathological alterations were most severe in the groups treated with higher doses of metal mixture. Similarly, the brain acetylcholinesterase (AChE) activity demonstrated a dose-dependent decrease significant in higher doses, while BMDI suggested Cd as the main contributor in the examined metal mixture. These findings imply varying susceptibility of neurotoxic endpoints to different doses of environmentally relevant metal mixtures, advocating for risk assessment and regulatory measures to address metal pollution and enhance remediation strategies.
PB  - Elsevier Inc.
T2  - Environmental Research
T1  - Neurotoxic effects of low dose ranges of environmental metal mixture in a rat model: The benchmark approach
VL  - 252
IS  - 1
DO  - 10.1016/j.envres.2024.118680
ER  - 

@article{
author = "Živančević, Katarina and Baralić, Katarina and Vukelić, Dragana and Marić, Đurđica and Kotur-Stevuljević, Jelena and Ivanišević, Jasmina and Savić, Miroslav and Batinić, Bojan and Janković, Radmila and Buha-Đorđević, Aleksandra and Antonijević-Miljaković, Evica and Ćurčić, Marijana and Bulat, Zorica and Antonijević, Biljana and Đukić-Ćosić, Danijela",
year = "2024",
abstract = "Metals exert detrimental effects on various systems within the body, including the nervous system. Nevertheless, the dose-response relationship concerning the administration of low doses of metal mixtures remains inadequately explored. The assessment of neurotoxic effects of lead, cadmium, mercury, and arsenic mixture (MIX) administered at low dose ranges, was conducted using an in vivo approach. A subacute study was conducted on a rat model consisting of a control and five treatment groups subjected to oral exposure with gradually increasing doses (from MIX 1 to MIX 5). The results indicated that behavioural patterns in an already developed nervous system displayed a reduced susceptibility to the metal mixture exposure with tendency of higher doses to alter short term memory. However, the vulnerability of the mature brain to even minimal amounts of the investigated metal mixture was evident, particularly in the context of oxidative stress. Moreover, the study highlights superoxide dismutase's sensitivity as an early-stage neurotoxicity marker, as indicated by dose-dependent induction of oxidative stress in the brain revealed through Benchmark analysis. The narrowest Benchmark Dose Interval (BMDI) for superoxide dismutase (SOD) activity (1e-06 - 3.18e-05 mg As/kg b.w./day) indicates that arsenic may dictate the alterations in SOD activity when co-exposed with the other examined metals. The predicted Benchmark doses for oxidative stress parameters were very low, supporting “no-threshold” concept. Histopathological alterations were most severe in the groups treated with higher doses of metal mixture. Similarly, the brain acetylcholinesterase (AChE) activity demonstrated a dose-dependent decrease significant in higher doses, while BMDI suggested Cd as the main contributor in the examined metal mixture. These findings imply varying susceptibility of neurotoxic endpoints to different doses of environmentally relevant metal mixtures, advocating for risk assessment and regulatory measures to address metal pollution and enhance remediation strategies.",
publisher = "Elsevier Inc.",
journal = "Environmental Research",
title = "Neurotoxic effects of low dose ranges of environmental metal mixture in a rat model: The benchmark approach",
volume = "252",
number = "1",
doi = "10.1016/j.envres.2024.118680"
}

Živančević, K., Baralić, K., Vukelić, D., Marić, Đ., Kotur-Stevuljević, J., Ivanišević, J., Savić, M., Batinić, B., Janković, R., Buha-Đorđević, A., Antonijević-Miljaković, E., Ćurčić, M., Bulat, Z., Antonijević, B.,& Đukić-Ćosić, D.. (2024). Neurotoxic effects of low dose ranges of environmental metal mixture in a rat model: The benchmark approach. in Environmental Research
Elsevier Inc.., 252(1).
https://doi.org/10.1016/j.envres.2024.118680

Živančević K, Baralić K, Vukelić D, Marić Đ, Kotur-Stevuljević J, Ivanišević J, Savić M, Batinić B, Janković R, Buha-Đorđević A, Antonijević-Miljaković E, Ćurčić M, Bulat Z, Antonijević B, Đukić-Ćosić D. Neurotoxic effects of low dose ranges of environmental metal mixture in a rat model: The benchmark approach. in Environmental Research. 2024;252(1).
doi:10.1016/j.envres.2024.118680 .

Živančević, Katarina, Baralić, Katarina, Vukelić, Dragana, Marić, Đurđica, Kotur-Stevuljević, Jelena, Ivanišević, Jasmina, Savić, Miroslav, Batinić, Bojan, Janković, Radmila, Buha-Đorđević, Aleksandra, Antonijević-Miljaković, Evica, Ćurčić, Marijana, Bulat, Zorica, Antonijević, Biljana, Đukić-Ćosić, Danijela, "Neurotoxic effects of low dose ranges of environmental metal mixture in a rat model: The benchmark approach" in Environmental Research, 252, no. 1 (2024),
https://doi.org/10.1016/j.envres.2024.118680 . .

TY  - JOUR
AU  - Sharmin, Dishary
AU  - Divović, Branka
AU  - Ping, Xingjie
AU  - Cerne, Rok
AU  - Smith, Jodi L.
AU  - Rezvanian, Sepideh
AU  - Mondal, Prithu
AU  - Michelle, Meyer Jean
AU  - Kiley, Molly E.
AU  - Arnold, Leggy A.
AU  - Mian, Md Yeunus
AU  - Pandey, Kamal P.
AU  - Jin, Xiaoming
AU  - Mitrović, Jelena
AU  - Đorović, Đorđe
AU  - Lippa, Arnold
AU  - Cook, James M.
AU  - Golani, Lalit K.
AU  - Scholze, Petra
AU  - Savić, Miroslav
AU  - Witkin, Jeffrey M.
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5505
AB  - KRM-II-81 (1) is an imidazodiazepine GABAA receptor (GABAAR) potentiator with broad antiseizure efficacy and a low sedative burden. A brominated analogue, DS-II-73 (5), was synthesized and pharmacologically characterized as a potential backup compound as KRM-II-81 moves forward into development. The synthesis from 2-amino-5-bromophenyl)(pyridin-2yl)methanone (6) was processed in five steps with an overall yield of 38% and without the need for a palladium catalyst. GABAAR binding occurred with a Ki of 150 nM, and only 3 of 41 screened binding sites produced inhibition ≥50% at 10 μM, and the potency to induce cytotoxicity was ≥240 mM. DS-II-73 was selective for α2/3/5- over that of α1-containing GABAARs. Oral exposure of plasma and brain of rats was more than sufficient to functionally impact GABAARs. Tonic convulsions in mice and lethality induced by pentylenetetrazol were suppressed by DS-II-73 after oral administration and latencies to clonic and tonic seizures were prolonged. Cortical slice preparations from a patient with pharmacoresistant epilepsy (mesial temporal lobe) showed decreases in the frequency of local field potentials by DS-II-73. As with KRM-II-81, the motor-impairing effects of DS-II-73 were low compared to diazepam. Molecular docking studies of DS-II-73 with the α1β3γ2L-configured GABAAR showed low interaction with α1His102 that is suggested as a potential molecular mechanism for its low sedative side effects. These findings support the viability of DS-II-73 as a backup molecule for its ethynyl analogue, KRM-II-81, with the human tissue data providing translational credibility.
PB  - American Chemical Society
T2  - ACS Chemical Neuroscience
T1  - New Imidazodiazepine Analogue, 5-(8-Bromo-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)oxazole, Provides a Simplified Synthetic Scheme, High Oral Plasma and Brain Exposures, and Produces Antiseizure Efficacy in Mice, and Antiepileptogenic Activity in Neural Networks in Brain Slices from a Patient with Mesial Temporal Lobe Epilepsy
VL  - 15
IS  - 3
SP  - 517
EP  - 526
DO  - 10.1021/acschemneuro.3c00555
ER  - 

@article{
author = "Sharmin, Dishary and Divović, Branka and Ping, Xingjie and Cerne, Rok and Smith, Jodi L. and Rezvanian, Sepideh and Mondal, Prithu and Michelle, Meyer Jean and Kiley, Molly E. and Arnold, Leggy A. and Mian, Md Yeunus and Pandey, Kamal P. and Jin, Xiaoming and Mitrović, Jelena and Đorović, Đorđe and Lippa, Arnold and Cook, James M. and Golani, Lalit K. and Scholze, Petra and Savić, Miroslav and Witkin, Jeffrey M.",
year = "2024",
abstract = "KRM-II-81 (1) is an imidazodiazepine GABAA receptor (GABAAR) potentiator with broad antiseizure efficacy and a low sedative burden. A brominated analogue, DS-II-73 (5), was synthesized and pharmacologically characterized as a potential backup compound as KRM-II-81 moves forward into development. The synthesis from 2-amino-5-bromophenyl)(pyridin-2yl)methanone (6) was processed in five steps with an overall yield of 38% and without the need for a palladium catalyst. GABAAR binding occurred with a Ki of 150 nM, and only 3 of 41 screened binding sites produced inhibition ≥50% at 10 μM, and the potency to induce cytotoxicity was ≥240 mM. DS-II-73 was selective for α2/3/5- over that of α1-containing GABAARs. Oral exposure of plasma and brain of rats was more than sufficient to functionally impact GABAARs. Tonic convulsions in mice and lethality induced by pentylenetetrazol were suppressed by DS-II-73 after oral administration and latencies to clonic and tonic seizures were prolonged. Cortical slice preparations from a patient with pharmacoresistant epilepsy (mesial temporal lobe) showed decreases in the frequency of local field potentials by DS-II-73. As with KRM-II-81, the motor-impairing effects of DS-II-73 were low compared to diazepam. Molecular docking studies of DS-II-73 with the α1β3γ2L-configured GABAAR showed low interaction with α1His102 that is suggested as a potential molecular mechanism for its low sedative side effects. These findings support the viability of DS-II-73 as a backup molecule for its ethynyl analogue, KRM-II-81, with the human tissue data providing translational credibility.",
publisher = "American Chemical Society",
journal = "ACS Chemical Neuroscience",
title = "New Imidazodiazepine Analogue, 5-(8-Bromo-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)oxazole, Provides a Simplified Synthetic Scheme, High Oral Plasma and Brain Exposures, and Produces Antiseizure Efficacy in Mice, and Antiepileptogenic Activity in Neural Networks in Brain Slices from a Patient with Mesial Temporal Lobe Epilepsy",
volume = "15",
number = "3",
pages = "517-526",
doi = "10.1021/acschemneuro.3c00555"
}

Sharmin, D., Divović, B., Ping, X., Cerne, R., Smith, J. L., Rezvanian, S., Mondal, P., Michelle, M. J., Kiley, M. E., Arnold, L. A., Mian, M. Y., Pandey, K. P., Jin, X., Mitrović, J., Đorović, Đ., Lippa, A., Cook, J. M., Golani, L. K., Scholze, P., Savić, M.,& Witkin, J. M.. (2024). New Imidazodiazepine Analogue, 5-(8-Bromo-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)oxazole, Provides a Simplified Synthetic Scheme, High Oral Plasma and Brain Exposures, and Produces Antiseizure Efficacy in Mice, and Antiepileptogenic Activity in Neural Networks in Brain Slices from a Patient with Mesial Temporal Lobe Epilepsy. in ACS Chemical Neuroscience
American Chemical Society., 15(3), 517-526.
https://doi.org/10.1021/acschemneuro.3c00555

Sharmin D, Divović B, Ping X, Cerne R, Smith JL, Rezvanian S, Mondal P, Michelle MJ, Kiley ME, Arnold LA, Mian MY, Pandey KP, Jin X, Mitrović J, Đorović Đ, Lippa A, Cook JM, Golani LK, Scholze P, Savić M, Witkin JM. New Imidazodiazepine Analogue, 5-(8-Bromo-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)oxazole, Provides a Simplified Synthetic Scheme, High Oral Plasma and Brain Exposures, and Produces Antiseizure Efficacy in Mice, and Antiepileptogenic Activity in Neural Networks in Brain Slices from a Patient with Mesial Temporal Lobe Epilepsy. in ACS Chemical Neuroscience. 2024;15(3):517-526.
doi:10.1021/acschemneuro.3c00555 .

Sharmin, Dishary, Divović, Branka, Ping, Xingjie, Cerne, Rok, Smith, Jodi L., Rezvanian, Sepideh, Mondal, Prithu, Michelle, Meyer Jean, Kiley, Molly E., Arnold, Leggy A., Mian, Md Yeunus, Pandey, Kamal P., Jin, Xiaoming, Mitrović, Jelena, Đorović, Đorđe, Lippa, Arnold, Cook, James M., Golani, Lalit K., Scholze, Petra, Savić, Miroslav, Witkin, Jeffrey M., "New Imidazodiazepine Analogue, 5-(8-Bromo-6-(pyridin-2-yl)-4H-benzo[f]imidazo[1,5-a][1,4]diazepin-3-yl)oxazole, Provides a Simplified Synthetic Scheme, High Oral Plasma and Brain Exposures, and Produces Antiseizure Efficacy in Mice, and Antiepileptogenic Activity in Neural Networks in Brain Slices from a Patient with Mesial Temporal Lobe Epilepsy" in ACS Chemical Neuroscience, 15, no. 3 (2024):517-526,
https://doi.org/10.1021/acschemneuro.3c00555 . .

TY  - GEN
AU  - Stanković, Tijana
AU  - Ilić, Tanja
AU  - Petković, Miloš
AU  - Pantelić, Ivana
AU  - Dobričić, Vladimir
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5578
AB  - The low aqueous and oil solubility of the novelpyrazoloquinolinone ligand (CW-02-79) with significantbinding affinity for sigma-2 receptors in the brain hindersthe development of conventional parenteral formulationsand thus a comprehensive evaluation of its efficacy andsafety. ...
T1  - Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5578
ER  - 

@misc{
author = "Stanković, Tijana and Ilić, Tanja and Petković, Miloš and Pantelić, Ivana and Dobričić, Vladimir and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2024",
abstract = "The low aqueous and oil solubility of the novelpyrazoloquinolinone ligand (CW-02-79) with significantbinding affinity for sigma-2 receptors in the brain hindersthe development of conventional parenteral formulationsand thus a comprehensive evaluation of its efficacy andsafety. ...",
title = "Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5578"
}

Stanković, T., Ilić, T., Petković, M., Pantelić, I., Dobričić, V., Cook, J. M., Savić, M.,& Savić, S.. (2024). Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation. .
https://hdl.handle.net/21.15107/rcub_farfar_5578

Stanković T, Ilić T, Petković M, Pantelić I, Dobričić V, Cook JM, Savić M, Savić S. Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation. 2024;.
https://hdl.handle.net/21.15107/rcub_farfar_5578 .

Stanković, Tijana, Ilić, Tanja, Petković, Miloš, Pantelić, Ivana, Dobričić, Vladimir, Cook, James M., Savić, Miroslav, Savić, Snežana, "Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation" (2024),
https://hdl.handle.net/21.15107/rcub_farfar_5578 .

TY  - CONF
AU  - Stanković, Tijana
AU  - Ilić, Tanja
AU  - Petković, Miloš
AU  - Pantelić, Ivana
AU  - Dobričić, Vladimir
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5577
AB  - The low aqueous and oil solubility of the novel
pyrazoloquinolinone ligand (CW-02-79) with significant
binding affinity for sigma-2 receptors in the brain hinders
the development of conventional parenteral formulations
and thus a comprehensive evaluation of its efficacy and
safety. ...
PB  - International Society of Drug Delivery Sciences and Technology (APGI)
PB  - International Association for Pharmaceutical Technology (APV)
PB  - Italian Society of Technology and Legislation (S.T.E.L.F)
C3  - 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria
T1  - Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5577
ER  - 

@conference{
author = "Stanković, Tijana and Ilić, Tanja and Petković, Miloš and Pantelić, Ivana and Dobričić, Vladimir and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2024",
abstract = "The low aqueous and oil solubility of the novel
pyrazoloquinolinone ligand (CW-02-79) with significant
binding affinity for sigma-2 receptors in the brain hinders
the development of conventional parenteral formulations
and thus a comprehensive evaluation of its efficacy and
safety. ...",
publisher = "International Society of Drug Delivery Sciences and Technology (APGI), International Association for Pharmaceutical Technology (APV), Italian Society of Technology and Legislation (S.T.E.L.F)",
journal = "14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria",
title = "Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5577"
}

Stanković, T., Ilić, T., Petković, M., Pantelić, I., Dobričić, V., Cook, J. M., Savić, M.,& Savić, S.. (2024). Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation. in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria
International Society of Drug Delivery Sciences and Technology (APGI)..
https://hdl.handle.net/21.15107/rcub_farfar_5577

Stanković T, Ilić T, Petković M, Pantelić I, Dobričić V, Cook JM, Savić M, Savić S. Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation. in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria. 2024;.
https://hdl.handle.net/21.15107/rcub_farfar_5577 .

Stanković, Tijana, Ilić, Tanja, Petković, Miloš, Pantelić, Ivana, Dobričić, Vladimir, Cook, James M., Savić, Miroslav, Savić, Snežana, "Overcoming the low solubility of novel pyrazoloquinolinone ligand (CW-02-79) by combination of drug-phospholipid complex and nanoemulsion technology: design and physicochemical evaluation" in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria (2024),
https://hdl.handle.net/21.15107/rcub_farfar_5577 .

TY  - CONF
AU  - Đoković, Jelena
AU  - Nikolić, Ines
AU  - Sharmin, Dishary
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Borchard, Gerrit
AU  - Savić, Snežana
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5575
AB  - INTRODUCTION Temporal lobe epilepsy is characterized by seizures, but can also be associated with mental health problems for which there are no clear treatment regimens. A proprietary compound, GL-II-73 - (4R)-8-ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxamide, a positive allosteric modulator of α5-containing γ-aminobutyric acid (GABA) receptors, has been shown to be effective in the treatment of these comorbidities [1]. ...
PB  - International Society of Drug Delivery Sciences and Technology (APGI)
PB  - International Association for Pharmaceutical Technology (APV)
PB  - Italian Society of Technology and Legislation (S.T.E.L.F)
C3  - 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria
T1  - The impact of the aqueous phase on the physicochemical characteristics of nanoemulsions loaded with patent protected compound GL-II-73
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5575
ER  - 

@conference{
author = "Đoković, Jelena and Nikolić, Ines and Sharmin, Dishary and Cook, James M. and Savić, Miroslav and Borchard, Gerrit and Savić, Snežana",
year = "2024",
abstract = "INTRODUCTION Temporal lobe epilepsy is characterized by seizures, but can also be associated with mental health problems for which there are no clear treatment regimens. A proprietary compound, GL-II-73 - (4R)-8-ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-benzo[f]imidazo[1,5-a][1,4]diazepine-3-carboxamide, a positive allosteric modulator of α5-containing γ-aminobutyric acid (GABA) receptors, has been shown to be effective in the treatment of these comorbidities [1]. ...",
publisher = "International Society of Drug Delivery Sciences and Technology (APGI), International Association for Pharmaceutical Technology (APV), Italian Society of Technology and Legislation (S.T.E.L.F)",
journal = "14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria",
title = "The impact of the aqueous phase on the physicochemical characteristics of nanoemulsions loaded with patent protected compound GL-II-73",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5575"
}

Đoković, J., Nikolić, I., Sharmin, D., Cook, J. M., Savić, M., Borchard, G.,& Savić, S.. (2024). The impact of the aqueous phase on the physicochemical characteristics of nanoemulsions loaded with patent protected compound GL-II-73. in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria
International Society of Drug Delivery Sciences and Technology (APGI)..
https://hdl.handle.net/21.15107/rcub_farfar_5575

Đoković J, Nikolić I, Sharmin D, Cook JM, Savić M, Borchard G, Savić S. The impact of the aqueous phase on the physicochemical characteristics of nanoemulsions loaded with patent protected compound GL-II-73. in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria. 2024;.
https://hdl.handle.net/21.15107/rcub_farfar_5575 .

Đoković, Jelena, Nikolić, Ines, Sharmin, Dishary, Cook, James M., Savić, Miroslav, Borchard, Gerrit, Savić, Snežana, "The impact of the aqueous phase on the physicochemical characteristics of nanoemulsions loaded with patent protected compound GL-II-73" in 14th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, 18 - 21 March 2024, Vienna, Austria (2024),
https://hdl.handle.net/21.15107/rcub_farfar_5575 .

TY  - JOUR
AU  - Radojević, Branislava S.
AU  - Jančić, Ivan
AU  - Savić, Miroslav
AU  - Kostić, Vladimir S.
AU  - Dragašević-Mišković, Nataša T.
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5581
AB  - Catechol-O-methyltransferase (COMT) is one of the cardinal enzymes in the degradation
of catecholamines and levodopa. Genetic variants of the COMT gene may affect COMT enzyme
activity. The most examined COMT gene polymorphism is the nonsynonymous single nucleotide
polymorphism (SNP) in exon 4 (Val108/158Met; rs4680). This highly functional polymorphism
is responsible for fourfold variations in enzyme activity and dopamine catabolism. Recent data
suggested that even synonymous SNPs of the COMT gene can lead to changes in enzyme activity.
Genetically determined COMT activity can affect an individual's response to levodopa therapy
and carries the risk of complications from prolonged levodopa use in Parkinson's disease (PD)
patients. Identifying at-risk individuals through genetic susceptibility markers could help to
prevent the development of levodopa-induced complications in PD.
AB  - Katehol-O-metiltransferaza (engl. catechol-O-methyltransferase, COMT) je jedan od
glavnih enzima u razgradnji kateholamina i levodope. Genetske varijante COMT gena mogu
uticati na aktivnost COMT enzima. Polimorfizam COMT gena koji je najviše proučavan je
nesinonimni jednonukleotidni polimorfizam (engl. single nucleotide polymorphism, SNP) u
egzonu 4 (Val108/158Met; rs4680). Ovaj visoko funkcionalni polimorfizam odgovoran je za
četvorostruke varijacije u aktivnosti enzima i katabolizmu dopamina. Nedavni podaci sugerišu da
čak i sinonimni SNP COMT gena mogu da dovedu do promena u aktivnosti enzima. Genetski
određene razlike u COMT aktivnosti mogu uticati na odgovor pojedinca na terapiju levodopom i
nose rizik od komplikacija dugotrajne primene levodope kod pacijenata sa Parkinsonovom
bolešću (PB). Identifikacija osoba u riziku putem markera genetske osetljivosti može pomoći u
prevenciji komplikacija izazvanih levodopom kod PB.
PB  - Beograd : Savez farmaceutskih udruženja Srbije
PB  - Beograd : Univerzitet u Beogradu - Farmaceutski fakultet
T2  - Arhiv za farmaciju
T1  - Association of catechol-O-methyltransferase gene polymorphisms with treatment response and levodopa-induced complications in Parkinson's disease: A summary of current knowledge
T1  - Udruženost polimorfizama gena za katehol-O- metiltransferazu sa terapijskim odgovorom i komplikacijama izazvanim levodopom kod Parkinsonove bolesti: Rezime sadašnjih saznanja
VL  - 74
IS  - 1
SP  - 23
EP  - 37
DO  - 10.5937/arhfarm74-45472
ER  - 

@article{
author = "Radojević, Branislava S. and Jančić, Ivan and Savić, Miroslav and Kostić, Vladimir S. and Dragašević-Mišković, Nataša T.",
year = "2024",
abstract = "Catechol-O-methyltransferase (COMT) is one of the cardinal enzymes in the degradation
of catecholamines and levodopa. Genetic variants of the COMT gene may affect COMT enzyme
activity. The most examined COMT gene polymorphism is the nonsynonymous single nucleotide
polymorphism (SNP) in exon 4 (Val108/158Met; rs4680). This highly functional polymorphism
is responsible for fourfold variations in enzyme activity and dopamine catabolism. Recent data
suggested that even synonymous SNPs of the COMT gene can lead to changes in enzyme activity.
Genetically determined COMT activity can affect an individual's response to levodopa therapy
and carries the risk of complications from prolonged levodopa use in Parkinson's disease (PD)
patients. Identifying at-risk individuals through genetic susceptibility markers could help to
prevent the development of levodopa-induced complications in PD., Katehol-O-metiltransferaza (engl. catechol-O-methyltransferase, COMT) je jedan od
glavnih enzima u razgradnji kateholamina i levodope. Genetske varijante COMT gena mogu
uticati na aktivnost COMT enzima. Polimorfizam COMT gena koji je najviše proučavan je
nesinonimni jednonukleotidni polimorfizam (engl. single nucleotide polymorphism, SNP) u
egzonu 4 (Val108/158Met; rs4680). Ovaj visoko funkcionalni polimorfizam odgovoran je za
četvorostruke varijacije u aktivnosti enzima i katabolizmu dopamina. Nedavni podaci sugerišu da
čak i sinonimni SNP COMT gena mogu da dovedu do promena u aktivnosti enzima. Genetski
određene razlike u COMT aktivnosti mogu uticati na odgovor pojedinca na terapiju levodopom i
nose rizik od komplikacija dugotrajne primene levodope kod pacijenata sa Parkinsonovom
bolešću (PB). Identifikacija osoba u riziku putem markera genetske osetljivosti može pomoći u
prevenciji komplikacija izazvanih levodopom kod PB.",
publisher = "Beograd : Savez farmaceutskih udruženja Srbije, Beograd : Univerzitet u Beogradu - Farmaceutski fakultet",
journal = "Arhiv za farmaciju",
title = "Association of catechol-O-methyltransferase gene polymorphisms with treatment response and levodopa-induced complications in Parkinson's disease: A summary of current knowledge, Udruženost polimorfizama gena za katehol-O- metiltransferazu sa terapijskim odgovorom i komplikacijama izazvanim levodopom kod Parkinsonove bolesti: Rezime sadašnjih saznanja",
volume = "74",
number = "1",
pages = "23-37",
doi = "10.5937/arhfarm74-45472"
}

Radojević, B. S., Jančić, I., Savić, M., Kostić, V. S.,& Dragašević-Mišković, N. T.. (2024). Association of catechol-O-methyltransferase gene polymorphisms with treatment response and levodopa-induced complications in Parkinson's disease: A summary of current knowledge. in Arhiv za farmaciju
Beograd : Savez farmaceutskih udruženja Srbije., 74(1), 23-37.
https://doi.org/10.5937/arhfarm74-45472

Radojević BS, Jančić I, Savić M, Kostić VS, Dragašević-Mišković NT. Association of catechol-O-methyltransferase gene polymorphisms with treatment response and levodopa-induced complications in Parkinson's disease: A summary of current knowledge. in Arhiv za farmaciju. 2024;74(1):23-37.
doi:10.5937/arhfarm74-45472 .

Radojević, Branislava S., Jančić, Ivan, Savić, Miroslav, Kostić, Vladimir S., Dragašević-Mišković, Nataša T., "Association of catechol-O-methyltransferase gene polymorphisms with treatment response and levodopa-induced complications in Parkinson's disease: A summary of current knowledge" in Arhiv za farmaciju, 74, no. 1 (2024):23-37,
https://doi.org/10.5937/arhfarm74-45472 . .

TY  - JOUR
AU  - Đorović, Đorđe
AU  - Lazarević, Vesna
AU  - Aranđelović, Jovana
AU  - Stevanović, Vladimir
AU  - Paslawski, Wojciech
AU  - Zhang, Xiaoqun
AU  - Velimirović, Milica
AU  - Petronijević, Nataša
AU  - Puškaš, Laslo
AU  - Savić, Miroslav
AU  - Svenningsson, Per
PY  - 2024
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5503
AB  - Background: Early life stress is a major risk factor for later development of psychiatric disorders, including post-traumatic stress disorder (PTSD). An intricate relationship exists between various neurotransmitters (such as glutamate, norepinephrine or serotonin), calcium/calmodulin-dependent protein kinase II (CaMKII), as an important regulator of glutamatergic synaptic function, and PTSD. Here, we developed a double-hit model to investigate the interaction of maternal deprivation (MD) as an early life stress model and single prolonged stress (SPS) as a PTSD model at the behavioral and molecular levels. Methods: Male Wistar rats exposed to these stress paradigms were subjected to a comprehensive behavioral analysis. In hippocampal synaptosomes we investigated neurotransmitter release and glutamate concentration. The expression of CaMKII and the content of monoamines were determined in selected brain regions. Brain-derived neurotrophic factor (BDNF) mRNA was quantified by radioactive in situ hybridization. Results: We report a distinct behavioral phenotype in the double-hit group. Double-hit and SPS groups had decreased hippocampal presynaptic glutamatergic function. In hippocampus, double-hit stress caused a decrease in autophosphorylation of CaMKII. In prefrontal cortex, both SPS and double-hit stress had a similar effect on CaMKII autophosphorylation. Double-hit stress, rather than SPS, affected the norepinephrine and serotonin levels in prefrontal cortex, and suppressed BDNF gene expression in prefrontal cortex and hippocampus. Limitations: The study was conducted in male rats only. The affected brain regions cannot be restricted to hippocampus, prefrontal cortex and amygdala. Conclusion: Double-hit stress caused more pronounced and distinct behavioral, molecular and functional changes, compared to MD or SPS alone.
PB  - Elsevier B.V.
T2  - Journal of Affective Disorders
T1  - Maternal deprivation causes CaMKII downregulation and modulates glutamate, norepinephrine and serotonin in limbic brain areas in a rat model of single prolonged stress
VL  - 349
SP  - 286
EP  - 296
DO  - 10.1016/j.jad.2024.01.087
ER  - 

@article{
author = "Đorović, Đorđe and Lazarević, Vesna and Aranđelović, Jovana and Stevanović, Vladimir and Paslawski, Wojciech and Zhang, Xiaoqun and Velimirović, Milica and Petronijević, Nataša and Puškaš, Laslo and Savić, Miroslav and Svenningsson, Per",
year = "2024",
abstract = "Background: Early life stress is a major risk factor for later development of psychiatric disorders, including post-traumatic stress disorder (PTSD). An intricate relationship exists between various neurotransmitters (such as glutamate, norepinephrine or serotonin), calcium/calmodulin-dependent protein kinase II (CaMKII), as an important regulator of glutamatergic synaptic function, and PTSD. Here, we developed a double-hit model to investigate the interaction of maternal deprivation (MD) as an early life stress model and single prolonged stress (SPS) as a PTSD model at the behavioral and molecular levels. Methods: Male Wistar rats exposed to these stress paradigms were subjected to a comprehensive behavioral analysis. In hippocampal synaptosomes we investigated neurotransmitter release and glutamate concentration. The expression of CaMKII and the content of monoamines were determined in selected brain regions. Brain-derived neurotrophic factor (BDNF) mRNA was quantified by radioactive in situ hybridization. Results: We report a distinct behavioral phenotype in the double-hit group. Double-hit and SPS groups had decreased hippocampal presynaptic glutamatergic function. In hippocampus, double-hit stress caused a decrease in autophosphorylation of CaMKII. In prefrontal cortex, both SPS and double-hit stress had a similar effect on CaMKII autophosphorylation. Double-hit stress, rather than SPS, affected the norepinephrine and serotonin levels in prefrontal cortex, and suppressed BDNF gene expression in prefrontal cortex and hippocampus. Limitations: The study was conducted in male rats only. The affected brain regions cannot be restricted to hippocampus, prefrontal cortex and amygdala. Conclusion: Double-hit stress caused more pronounced and distinct behavioral, molecular and functional changes, compared to MD or SPS alone.",
publisher = "Elsevier B.V.",
journal = "Journal of Affective Disorders",
title = "Maternal deprivation causes CaMKII downregulation and modulates glutamate, norepinephrine and serotonin in limbic brain areas in a rat model of single prolonged stress",
volume = "349",
pages = "286-296",
doi = "10.1016/j.jad.2024.01.087"
}

Đorović, Đ., Lazarević, V., Aranđelović, J., Stevanović, V., Paslawski, W., Zhang, X., Velimirović, M., Petronijević, N., Puškaš, L., Savić, M.,& Svenningsson, P.. (2024). Maternal deprivation causes CaMKII downregulation and modulates glutamate, norepinephrine and serotonin in limbic brain areas in a rat model of single prolonged stress. in Journal of Affective Disorders
Elsevier B.V.., 349, 286-296.
https://doi.org/10.1016/j.jad.2024.01.087

Đorović Đ, Lazarević V, Aranđelović J, Stevanović V, Paslawski W, Zhang X, Velimirović M, Petronijević N, Puškaš L, Savić M, Svenningsson P. Maternal deprivation causes CaMKII downregulation and modulates glutamate, norepinephrine and serotonin in limbic brain areas in a rat model of single prolonged stress. in Journal of Affective Disorders. 2024;349:286-296.
doi:10.1016/j.jad.2024.01.087 .

Đorović, Đorđe, Lazarević, Vesna, Aranđelović, Jovana, Stevanović, Vladimir, Paslawski, Wojciech, Zhang, Xiaoqun, Velimirović, Milica, Petronijević, Nataša, Puškaš, Laslo, Savić, Miroslav, Svenningsson, Per, "Maternal deprivation causes CaMKII downregulation and modulates glutamate, norepinephrine and serotonin in limbic brain areas in a rat model of single prolonged stress" in Journal of Affective Disorders, 349 (2024):286-296,
https://doi.org/10.1016/j.jad.2024.01.087 . .

TY  - JOUR
AU  - Pandey, Kamal P.
AU  - Divović, Branka
AU  - Rashid, Farjana
AU  - Golani, Lalit K.
AU  - Cerne, Rok
AU  - Zahn, Nicolas M.
AU  - Meyer, Michelle Jean
AU  - Arnold, Leggy A.
AU  - Sharmin, Dishary
AU  - Mian, Md Yeunus
AU  - Smith, Jodi L.
AU  - Ping, Xingjie
AU  - Jin, Xiaoming
AU  - Lippa, Arnold
AU  - Tiruveedhula, Phani Babu V. V. N.
AU  - Cook, James M.
AU  - Savić, Miroslav
AU  - Witkin, Jeffrey M.
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5605
AB  - To provide back-up compounds to support the development of the GABAA receptor (GABAAR) potentiator KRM-II-81, three novel analogs were designed: replacing the pyridinyl with 2'-Cl-phenyl (FR-II-60), changing the positions of the N and O atoms in the oxazole ring with addition of an ethyl group (KPPIII-34 and KPP-III-51), or substituting a Br atom for the ethynyl of KRM-II-81 (KPP-III-34). The compounds bound to brain GABAARs. Intraperitoneal administration of FR-II-60 and KPP-III-34 produced anticonvulsant activity in mice [maximal electroshock (MES)-induced seizures or 6 Hz-induced seizures], whereas KPPIII-51 did not. Although all compounds were orally bioavailable, structural changes reduced the plasma and brain (FR-II-60 and KPP-III-51) exposures relative to KRM-II-81. Oral administration of each compound produced dose-dependent increases in the latency for both clonic and tonic seizures and the lethality induced by pentylenetetrazol (PTZ) in mice. Since KPP-III-34 produced the highest brain area under the curve (AUC) exposures, it was selected for further profiling. Oral administration of KPP-III-34 suppressed seizures in corneal-kindled mice, hippocampal paroxysmal discharges in mesial temporal lobe epileptic mice, and PTZ-induced convulsions in rats. Only transient sensorimotor impairment was observed in mice, and doses of KPP-III-34 up to 500 mg/kg did not produce impairment in rats. Molecular docking studies demonstrated that all compounds displayed a reduced propensity for binding to a1His102 compared with the sedating compound alprazolam; the bromine-substituted KPP-III-34 achieved the least interaction. Overall, these findings document the oral bioavailability and anticonvulsant efficacy of three novel analogs of KRM-II-81 with reduced sedative effects. SIGNIFICANCE STATEMENT A new non-sedating compound, KRM-II-81, with reduced propensity for tolerance is moving into clinical development. Three new analogs were orally bioavailable, produced anticonvulsant effects in rodents, and displayed low sensorimotor impairment. KPP-III-34 demonstrated efficacy in models of pharmacoresistant epilepsy. Docking studies demonstrated a low propensity for compound binding to the a1His102 residue implicated in sedation. Thus, three additional structures have been added to the list of non-sedating imidazodiazepine anticonvulsants that could serve as backups in the clinical development of KRM-II-81. Copyright © 2023 by The American Society for Pharmacology and Experimental Therapeutics.
PB  - American Society for Pharmacology and Experimental Therapy (ASPET)
T2  - Journal of Pharmacology and Experimental Therapeutics
T1  - Structural Analogs of the GABAkine KRM-II-81 Are Orally Bioavailable Anticonvulsants without Sedation
VL  - 385
IS  - 1
SP  - 50
EP  - 61
DO  - 10.1124/jpet.122.001362
ER  - 

@article{
author = "Pandey, Kamal P. and Divović, Branka and Rashid, Farjana and Golani, Lalit K. and Cerne, Rok and Zahn, Nicolas M. and Meyer, Michelle Jean and Arnold, Leggy A. and Sharmin, Dishary and Mian, Md Yeunus and Smith, Jodi L. and Ping, Xingjie and Jin, Xiaoming and Lippa, Arnold and Tiruveedhula, Phani Babu V. V. N. and Cook, James M. and Savić, Miroslav and Witkin, Jeffrey M.",
year = "2023",
abstract = "To provide back-up compounds to support the development of the GABAA receptor (GABAAR) potentiator KRM-II-81, three novel analogs were designed: replacing the pyridinyl with 2'-Cl-phenyl (FR-II-60), changing the positions of the N and O atoms in the oxazole ring with addition of an ethyl group (KPPIII-34 and KPP-III-51), or substituting a Br atom for the ethynyl of KRM-II-81 (KPP-III-34). The compounds bound to brain GABAARs. Intraperitoneal administration of FR-II-60 and KPP-III-34 produced anticonvulsant activity in mice [maximal electroshock (MES)-induced seizures or 6 Hz-induced seizures], whereas KPPIII-51 did not. Although all compounds were orally bioavailable, structural changes reduced the plasma and brain (FR-II-60 and KPP-III-51) exposures relative to KRM-II-81. Oral administration of each compound produced dose-dependent increases in the latency for both clonic and tonic seizures and the lethality induced by pentylenetetrazol (PTZ) in mice. Since KPP-III-34 produced the highest brain area under the curve (AUC) exposures, it was selected for further profiling. Oral administration of KPP-III-34 suppressed seizures in corneal-kindled mice, hippocampal paroxysmal discharges in mesial temporal lobe epileptic mice, and PTZ-induced convulsions in rats. Only transient sensorimotor impairment was observed in mice, and doses of KPP-III-34 up to 500 mg/kg did not produce impairment in rats. Molecular docking studies demonstrated that all compounds displayed a reduced propensity for binding to a1His102 compared with the sedating compound alprazolam; the bromine-substituted KPP-III-34 achieved the least interaction. Overall, these findings document the oral bioavailability and anticonvulsant efficacy of three novel analogs of KRM-II-81 with reduced sedative effects. SIGNIFICANCE STATEMENT A new non-sedating compound, KRM-II-81, with reduced propensity for tolerance is moving into clinical development. Three new analogs were orally bioavailable, produced anticonvulsant effects in rodents, and displayed low sensorimotor impairment. KPP-III-34 demonstrated efficacy in models of pharmacoresistant epilepsy. Docking studies demonstrated a low propensity for compound binding to the a1His102 residue implicated in sedation. Thus, three additional structures have been added to the list of non-sedating imidazodiazepine anticonvulsants that could serve as backups in the clinical development of KRM-II-81. Copyright © 2023 by The American Society for Pharmacology and Experimental Therapeutics.",
publisher = "American Society for Pharmacology and Experimental Therapy (ASPET)",
journal = "Journal of Pharmacology and Experimental Therapeutics",
title = "Structural Analogs of the GABAkine KRM-II-81 Are Orally Bioavailable Anticonvulsants without Sedation",
volume = "385",
number = "1",
pages = "50-61",
doi = "10.1124/jpet.122.001362"
}

Pandey, K. P., Divović, B., Rashid, F., Golani, L. K., Cerne, R., Zahn, N. M., Meyer, M. J., Arnold, L. A., Sharmin, D., Mian, M. Y., Smith, J. L., Ping, X., Jin, X., Lippa, A., Tiruveedhula, P. B. V. V. N., Cook, J. M., Savić, M.,& Witkin, J. M.. (2023). Structural Analogs of the GABAkine KRM-II-81 Are Orally Bioavailable Anticonvulsants without Sedation. in Journal of Pharmacology and Experimental Therapeutics
American Society for Pharmacology and Experimental Therapy (ASPET)., 385(1), 50-61.
https://doi.org/10.1124/jpet.122.001362

Pandey KP, Divović B, Rashid F, Golani LK, Cerne R, Zahn NM, Meyer MJ, Arnold LA, Sharmin D, Mian MY, Smith JL, Ping X, Jin X, Lippa A, Tiruveedhula PBVVN, Cook JM, Savić M, Witkin JM. Structural Analogs of the GABAkine KRM-II-81 Are Orally Bioavailable Anticonvulsants without Sedation. in Journal of Pharmacology and Experimental Therapeutics. 2023;385(1):50-61.
doi:10.1124/jpet.122.001362 .

Pandey, Kamal P., Divović, Branka, Rashid, Farjana, Golani, Lalit K., Cerne, Rok, Zahn, Nicolas M., Meyer, Michelle Jean, Arnold, Leggy A., Sharmin, Dishary, Mian, Md Yeunus, Smith, Jodi L., Ping, Xingjie, Jin, Xiaoming, Lippa, Arnold, Tiruveedhula, Phani Babu V. V. N., Cook, James M., Savić, Miroslav, Witkin, Jeffrey M., "Structural Analogs of the GABAkine KRM-II-81 Are Orally Bioavailable Anticonvulsants without Sedation" in Journal of Pharmacology and Experimental Therapeutics, 385, no. 1 (2023):50-61,
https://doi.org/10.1124/jpet.122.001362 . .

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/123456789/5049
AB  - Introduction Nanopharmaceuticals offer a good method to avoid some of the difficulties that novel drug candidates confront. They can be tailored to adjust their water solubility, half-life, biodistribution, and govern the release of the integrated medication. Because of the excipients utilized, lipid nanocarriers (liposomes, nanoemulsions (NEs), nanoparticles) have been used to increase brain targeting (1,2). The investigated compound (GL-II-73) - (4R)-8-ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-benzo[f]imidazo[1,5-a] [1,4]diazepine-3-carboxamide is imidazobenzodiazepine (IBZD) ligand that acts as positive allosteric modulator on α-GABAA receptors and was shown to possess combined antidepressant and pro-cognitive effects, making it a promising candidate for further research (3). This work aims to investigate the physicochemical features of GL-II-73 to pick the best parenteral nanodelivery system for prospective research to assess its parameters. Мaterials and methods The saturation solubility of GL-II-73 was determined by adding it in excess to various oils (medium chain triglycerides, soybean, castor, and fish oil) to assess the oil solubility for the substance and select the optimal oil phase composition capable of incorporating the highest concentration of the GL-II-73. It was necessary to test the substance's solubility in buffers of various pH values to determine whether it has pH-dependent solubility. This investigation was carried out by incubating GL-II-73 and studied mediums on vortex for 24 hours and centrifuging to isolate supernatants from which the GL-II-73 concentration was evaluated using the LC-MS/MS method. The measurements were taken three times. In addition, after a 24-hour equilibration interval and determination of the GL-II-73 concentration, the log P value was obtained in an octanol/water system. Based on these findings, preliminary GL-II-73 (NE) was prepared using the high pressure homogenization method. In brief, the oil and aqueous phases were prepared separately and heated to 50 ˚C. They were then pre-mixed at the rotor stator homogenizer before being homogenized for 10 discontinuous cycles at 800 bar on the high pressure homogenizer. After diluting the sample in 1:500, v/v, ultra-purified water, the resulting formulations were characterized in terms of droplet size using the dynamic light scattering (DLS) technique on a Zetasizer Nano ZS90 (Malvern Instruments Ltd., Worcestershire, U.K.). On the same equipment, the NE's zeta potential (ZP) was measured. In addition, the pH and conductivity of the samples were examined. The ultrafiltration technique was used to evaluate the encapsulation efficacy (EE) by depositing 2 ml of the material in Amicon Ultra-4; NMWL 10 kDa filter units and centrifuging at 4500 rcf for 90 minutes. The EE was computed as %EE = ((A formulation A filtrate)/A formulation) 100, where A formulation represents the compound content in the formulation and A filtrate represents the filtrate, which was diluted in methanol and analyzed for Gl-II-73 content using the LC-MS/MS technique. During the one-month storage period, these conditions were monitored. Results and discussion Table 1 shows the solubility of GL-II-73 in the oils and buffers tested. The relatively good oil solubility, together with the log P value of 2.09, suggested that NEs could be promising carriers for GL-II-73. The highest oil solubility was detected in medium chain triglycerides, making them the oil phase of choice for future formulation development. Based on the solubility in 0.1 M HCl and phosphate buffer pH 7.4, it is possible to deduce that GL-II-73 has pH dependent solubility, with increased solubility observed as pH decreases, most likely due to the presence of ionizable functional groups and multiple H-bond acceptors. This suggested that the best EE would most likely be obtained by increasing the pH of the aqueous phase and keeping the chemical entrapped in the NE droplets. Solubility in organic solvents revealed that methanol is the best solvent for GL-II-73, as expected given its greater polarity index compared to isopropanol, which is why it was chosen for future characterization. Based on the solubility study, NE of the following composition was prepared: oil phase - medium chain tryglicerides (20%, w/w), soybean lecithin (2%, w/w), buthylhidroxytoluen (0.05%, w/w) and aqueous phase polysorbate 80 (2%, w/w), glycerol (2.25%, w/w), sodium oleate (0.03%, w/w), GL-II-73 (0.2%, w/w) and highly purified water to 100. The values of physicochemical parameters (Z-ave, PDI, ZP, pH, conductivity, drug content and encapsulation efficacy), measured both initially and after one month of storage, indicate suitability for parenteral administration. Conclusion Preliminary studies suggested that NEs are good prospective carriers for GL-II-73, but further research is needed for stability optimization.
PB  - Macedonian Pharmaceutical Association
PB  - Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy
C3  - 14th Central European Symposium on Pharmaceutical Technology, 28th - 30th September, Ohrid, N. Macedonia
T1  - Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5049
ER  - 

@conference{
author = "Đoković, Jelena and Marković, Bojan and Sharmin, Dishary and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Introduction Nanopharmaceuticals offer a good method to avoid some of the difficulties that novel drug candidates confront. They can be tailored to adjust their water solubility, half-life, biodistribution, and govern the release of the integrated medication. Because of the excipients utilized, lipid nanocarriers (liposomes, nanoemulsions (NEs), nanoparticles) have been used to increase brain targeting (1,2). The investigated compound (GL-II-73) - (4R)-8-ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-benzo[f]imidazo[1,5-a] [1,4]diazepine-3-carboxamide is imidazobenzodiazepine (IBZD) ligand that acts as positive allosteric modulator on α-GABAA receptors and was shown to possess combined antidepressant and pro-cognitive effects, making it a promising candidate for further research (3). This work aims to investigate the physicochemical features of GL-II-73 to pick the best parenteral nanodelivery system for prospective research to assess its parameters. Мaterials and methods The saturation solubility of GL-II-73 was determined by adding it in excess to various oils (medium chain triglycerides, soybean, castor, and fish oil) to assess the oil solubility for the substance and select the optimal oil phase composition capable of incorporating the highest concentration of the GL-II-73. It was necessary to test the substance's solubility in buffers of various pH values to determine whether it has pH-dependent solubility. This investigation was carried out by incubating GL-II-73 and studied mediums on vortex for 24 hours and centrifuging to isolate supernatants from which the GL-II-73 concentration was evaluated using the LC-MS/MS method. The measurements were taken three times. In addition, after a 24-hour equilibration interval and determination of the GL-II-73 concentration, the log P value was obtained in an octanol/water system. Based on these findings, preliminary GL-II-73 (NE) was prepared using the high pressure homogenization method. In brief, the oil and aqueous phases were prepared separately and heated to 50 ˚C. They were then pre-mixed at the rotor stator homogenizer before being homogenized for 10 discontinuous cycles at 800 bar on the high pressure homogenizer. After diluting the sample in 1:500, v/v, ultra-purified water, the resulting formulations were characterized in terms of droplet size using the dynamic light scattering (DLS) technique on a Zetasizer Nano ZS90 (Malvern Instruments Ltd., Worcestershire, U.K.). On the same equipment, the NE's zeta potential (ZP) was measured. In addition, the pH and conductivity of the samples were examined. The ultrafiltration technique was used to evaluate the encapsulation efficacy (EE) by depositing 2 ml of the material in Amicon Ultra-4; NMWL 10 kDa filter units and centrifuging at 4500 rcf for 90 minutes. The EE was computed as %EE = ((A formulation A filtrate)/A formulation) 100, where A formulation represents the compound content in the formulation and A filtrate represents the filtrate, which was diluted in methanol and analyzed for Gl-II-73 content using the LC-MS/MS technique. During the one-month storage period, these conditions were monitored. Results and discussion Table 1 shows the solubility of GL-II-73 in the oils and buffers tested. The relatively good oil solubility, together with the log P value of 2.09, suggested that NEs could be promising carriers for GL-II-73. The highest oil solubility was detected in medium chain triglycerides, making them the oil phase of choice for future formulation development. Based on the solubility in 0.1 M HCl and phosphate buffer pH 7.4, it is possible to deduce that GL-II-73 has pH dependent solubility, with increased solubility observed as pH decreases, most likely due to the presence of ionizable functional groups and multiple H-bond acceptors. This suggested that the best EE would most likely be obtained by increasing the pH of the aqueous phase and keeping the chemical entrapped in the NE droplets. Solubility in organic solvents revealed that methanol is the best solvent for GL-II-73, as expected given its greater polarity index compared to isopropanol, which is why it was chosen for future characterization. Based on the solubility study, NE of the following composition was prepared: oil phase - medium chain tryglicerides (20%, w/w), soybean lecithin (2%, w/w), buthylhidroxytoluen (0.05%, w/w) and aqueous phase polysorbate 80 (2%, w/w), glycerol (2.25%, w/w), sodium oleate (0.03%, w/w), GL-II-73 (0.2%, w/w) and highly purified water to 100. The values of physicochemical parameters (Z-ave, PDI, ZP, pH, conductivity, drug content and encapsulation efficacy), measured both initially and after one month of storage, indicate suitability for parenteral administration. Conclusion Preliminary studies suggested that NEs are good prospective carriers for GL-II-73, but further research is needed for stability optimization.",
publisher = "Macedonian Pharmaceutical Association, Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy",
journal = "14th Central European Symposium on Pharmaceutical Technology, 28th - 30th September, Ohrid, N. Macedonia",
title = "Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5049"
}

Đoković, J., Marković, B., Sharmin, D., Cook, J. M., Savić, M.,& Savić, S.. (2023). Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73. in 14th Central European Symposium on Pharmaceutical Technology, 28th - 30th September, Ohrid, N. Macedonia
Macedonian Pharmaceutical Association..
https://hdl.handle.net/21.15107/rcub_farfar_5049

Đoković J, Marković B, Sharmin D, Cook JM, Savić M, Savić S. Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73. in 14th Central European Symposium on Pharmaceutical Technology, 28th - 30th September, Ohrid, N. Macedonia. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5049 .

Đoković, Jelena, Marković, Bojan, Sharmin, Dishary, Cook, James M., Savić, Miroslav, Savić, Snežana, "Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73" in 14th Central European Symposium on Pharmaceutical Technology, 28th - 30th September, Ohrid, N. Macedonia (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5049 .

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  - 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  - 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/123456789/5050
AB  - Nanopharmaceuticals offer a good option to avoid
some of the difficulties that novel drug candidates
confront. They can be tailored to adjust their water
solubility, half-life, biodistribution, and govern the release
of the integrated medication. Because of the excipients
utilized, lipid nanocarriers (liposomes, nanoemulsions
(NEs), nanoparticles) have been used to increase brain
targeting (Bisso et al., 2020; Ilić et al., 2023).
The investigated compound (GL-II-73) - (4R)-8-
ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-
benzo[f]imidazo[1,5-a] [1,4]diazepine-3-carboxamide is
imidazobenzodiazepine (IBZD) ligand that acts as positive
allosteric modulator on α-GABAA receptors and was
shown to possess combined antidepressant and pro-
cognitive effects, making it a promising candidate for
further research (Prevot et al., 2019).
This work aims to investigate the physicochemical
features of GL-II-73 to pick the best parenteral
nanodelivery system for prospective research to assess its
parameters.
PB  - Macedonian Pharmaceutical Association
PB  - Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy
C3  - Macedonian Pharmaceutical Bulletin
T1  - Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73
VL  - 69
IS  - Suppl 1
SP  - 53
EP  - 54
DO  - 10.33320/maced.pharm.bull.2023.69.03.026
ER  - 

@conference{
author = "Đoković, Jelena and Marković, Bojan and Sharmin, Dishary and Cook, James M. and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Nanopharmaceuticals offer a good option to avoid
some of the difficulties that novel drug candidates
confront. They can be tailored to adjust their water
solubility, half-life, biodistribution, and govern the release
of the integrated medication. Because of the excipients
utilized, lipid nanocarriers (liposomes, nanoemulsions
(NEs), nanoparticles) have been used to increase brain
targeting (Bisso et al., 2020; Ilić et al., 2023).
The investigated compound (GL-II-73) - (4R)-8-
ethynyl-6-(2-fluorophenyl)-N,N,4-trimethyl-4H-
benzo[f]imidazo[1,5-a] [1,4]diazepine-3-carboxamide is
imidazobenzodiazepine (IBZD) ligand that acts as positive
allosteric modulator on α-GABAA receptors and was
shown to possess combined antidepressant and pro-
cognitive effects, making it a promising candidate for
further research (Prevot et al., 2019).
This work aims to investigate the physicochemical
features of GL-II-73 to pick the best parenteral
nanodelivery system for prospective research to assess its
parameters.",
publisher = "Macedonian Pharmaceutical Association, Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy",
journal = "Macedonian Pharmaceutical Bulletin",
title = "Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73",
volume = "69",
number = "Suppl 1",
pages = "53-54",
doi = "10.33320/maced.pharm.bull.2023.69.03.026"
}

Đoković, J., Marković, B., Sharmin, D., Cook, J. M., Savić, M.,& Savić, S.. (2023). Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73. in Macedonian Pharmaceutical Bulletin
Macedonian Pharmaceutical Association., 69(Suppl 1), 53-54.
https://doi.org/10.33320/maced.pharm.bull.2023.69.03.026

Đoković J, Marković B, Sharmin D, Cook JM, Savić M, Savić S. Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73. in Macedonian Pharmaceutical Bulletin. 2023;69(Suppl 1):53-54.
doi:10.33320/maced.pharm.bull.2023.69.03.026 .

Đoković, Jelena, Marković, Bojan, Sharmin, Dishary, Cook, James M., Savić, Miroslav, Savić, Snežana, "Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73" in Macedonian Pharmaceutical Bulletin, 69, no. Suppl 1 (2023):53-54,
https://doi.org/10.33320/maced.pharm.bull.2023.69.03.026 . .

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  - 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  - 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  - Stojanović, Goran M
AU  - Milić, Lazar
AU  - Endro, Ali A
AU  - Simić, Mitar
AU  - Nikolić, Ines
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5539
AB  - In this article, a sensing element for detection of date rape drugs in drinks is presented. The element consists of a three-dimensional printed holder, in which two embroidered electrodes have been fixed, forming a capacitive structure. As the dielectric properties of the liquid between these electrodes’ changes, the capacitance and consequently the impedance are changed. For experimental purposes, diazepam, a model rape drug, sucrose, and sodium chloride, which are used as control ingredients, were dissolved in a 40% V/V alcoholic beverage, serving as solutions for testing and comparison. The selectivity, repeatability, and sensitivity of the proposed sensor were tested. The sensitivity of detection of 1 mg/ml of diazepam in drinks was 0.92628 Ωl/mg, sucrose was 0.94774 Ωl/mg and sodium chloride was 2.46867 Ωl/mg, at 1 MHz. Moreover, with the use of the Cole impedance model, the selectivity of the sensor in the detection of diazepam, through the Nyquist plot and parameter estimation, has been achieved. Sensor repeatability was calculated through the relative standard deviation with the result for 1 mg/ml of diazepam dissolved in alcohol on 1 MHz being 2.48, in terms of impedance modulus. The presented sensor platform can successfully detect drugs in drinks and can protect from many cases of such assaults in a real world.
PB  - SAGE Publications Ltd
T2  - Textile Research Journal
T1  - Textile-based wearable device for detection of date rape drugs in drinks
DO  - 10.1177/00405175231218740
ER  - 

@article{
author = "Stojanović, Goran M and Milić, Lazar and Endro, Ali A and Simić, Mitar and Nikolić, Ines and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "In this article, a sensing element for detection of date rape drugs in drinks is presented. The element consists of a three-dimensional printed holder, in which two embroidered electrodes have been fixed, forming a capacitive structure. As the dielectric properties of the liquid between these electrodes’ changes, the capacitance and consequently the impedance are changed. For experimental purposes, diazepam, a model rape drug, sucrose, and sodium chloride, which are used as control ingredients, were dissolved in a 40% V/V alcoholic beverage, serving as solutions for testing and comparison. The selectivity, repeatability, and sensitivity of the proposed sensor were tested. The sensitivity of detection of 1 mg/ml of diazepam in drinks was 0.92628 Ωl/mg, sucrose was 0.94774 Ωl/mg and sodium chloride was 2.46867 Ωl/mg, at 1 MHz. Moreover, with the use of the Cole impedance model, the selectivity of the sensor in the detection of diazepam, through the Nyquist plot and parameter estimation, has been achieved. Sensor repeatability was calculated through the relative standard deviation with the result for 1 mg/ml of diazepam dissolved in alcohol on 1 MHz being 2.48, in terms of impedance modulus. The presented sensor platform can successfully detect drugs in drinks and can protect from many cases of such assaults in a real world.",
publisher = "SAGE Publications Ltd",
journal = "Textile Research Journal",
title = "Textile-based wearable device for detection of date rape drugs in drinks",
doi = "10.1177/00405175231218740"
}

Stojanović, G. M., Milić, L., Endro, A. A., Simić, M., Nikolić, I., Savić, M.,& Savić, S.. (2023). Textile-based wearable device for detection of date rape drugs in drinks. in Textile Research Journal
SAGE Publications Ltd..
https://doi.org/10.1177/00405175231218740

Stojanović GM, Milić L, Endro AA, Simić M, Nikolić I, Savić M, Savić S. Textile-based wearable device for detection of date rape drugs in drinks. in Textile Research Journal. 2023;.
doi:10.1177/00405175231218740 .

Stojanović, Goran M, Milić, Lazar, Endro, Ali A, Simić, Mitar, Nikolić, Ines, Savić, Miroslav, Savić, Snežana, "Textile-based wearable device for detection of date rape drugs in drinks" in Textile Research Journal (2023),
https://doi.org/10.1177/00405175231218740 . .

TY  - CONF
AU  - Aranđelović, Jovana
AU  - Kojić, Jana
AU  - Mirković, Kristina
AU  - Jančić, Ivan
AU  - Todorović, Lidija
AU  - Savić, Miroslav
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5520
AB  - Introduction: Impulsivity is an umbrella term that encompasses many subdomains, most of which rely on the decision-making processes. It is reported that in the process of healthy aging, the two dimensions of impulse control, cognition and motivation, are preserved or even improved. On the other hand, the attentive efficiency seems to decrease with age. Therefore, we aimed to investigate the effects of healthy aging on impulsivity in rats and the influence of food deprivation on impulsivity in aged rats as a strategy to enhance motivation. Additionally, we wanted to assess the gene expression for the alpha5 GABAA receptor subunit during aging, which plays a role in cognitive processes. Methodology: The variable-delay-to-signal (VDS) paradigm adapted to a touchscreen environment was used to assess impulsivity and attention in Sprague-Dawley rats at 2, 3, 5, 8, and 14 months of age. After one week of training, animals were tested at different ages in 3-stage testing protocol. Additionally, prior to testing, animals were fed a restricted diet (16 g/animal). The first stage included 20 trials with inter-trial interval of 6s (ITI6si) that reflected motor impulsivity. The second stage, with 60 randomly distributed trials of ITI9s or 15s, was related to delay intolerance, while the final stage (ITI6sf), similar to the first, was related to reflection impulsivity. The strict 3-day restriction diet (24h food deprivation followed by 10g/day/animal and 8g/day/animal) was applied to 14-month-old animals before testing. Gabra5 expression in the hippocampus was determined by qPCR. Results were analyzed by one-way ANOVA with or without repeated measures, followed by Sidak post-hoc test for impulsivity and attention parameters and by t-test for PCR parameters. Results: Animals aged 8 and 14 months had reduced motor impulsivity (p<0.01 for both groups) and delay intolerance (p<0.05 for both groups) and higher number of omissions (p<0.05 for both groups) compared to animals aged 2, 3 and 5 months of age. In addition, half of the animals were unable to successfully complete a task after 14 months. After rigorous food restriction in 14-month-old animals, the level of impulsivity (ITI9s and ITI15s) and attention (number of omissions) returned to the control level (2 and 3 months of age) compared to the performance of 14-month-old animals prior to rigorous food restriction (p<0.05). Further, the peak of reflection impulsivity (ITI6sf) was reached at 5 months compared to all other groups (p<0.01). No changes in Gabra5 expression in hippocampus were detected in 14-month-old compared to 3-month-old animals. Conclusion: From 8 months of age onwards, rats showed reduced impulsivity in the VDS stages where motor impulsivity and delay intolerance were tested, followed by attention deficits. After strict food restriction in 14-month-old animals, delay intolerance and attention were restored, suggesting the prominent role of motivation in controlling these processes, independently of Gabra5 expression levels in the hippocampus. Since the VDS paradigm aims to assess reward-related impulsivity based on cognition and motivation, it is suspected that results related to impaired cognition in older animals in other cognitive tests should be interpreted with caution, and with additional observation of motivation.
PB  - European College of Neuropsychopharmacology (ECNP)
C3  - 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain
T1  - Reward-related impulsivity as a possible surrogate marker of motivation in aging Sprague-Dawley rats
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5520
ER  - 

@conference{
author = "Aranđelović, Jovana and Kojić, Jana and Mirković, Kristina and Jančić, Ivan and Todorović, Lidija and Savić, Miroslav",
year = "2023",
abstract = "Introduction: Impulsivity is an umbrella term that encompasses many subdomains, most of which rely on the decision-making processes. It is reported that in the process of healthy aging, the two dimensions of impulse control, cognition and motivation, are preserved or even improved. On the other hand, the attentive efficiency seems to decrease with age. Therefore, we aimed to investigate the effects of healthy aging on impulsivity in rats and the influence of food deprivation on impulsivity in aged rats as a strategy to enhance motivation. Additionally, we wanted to assess the gene expression for the alpha5 GABAA receptor subunit during aging, which plays a role in cognitive processes. Methodology: The variable-delay-to-signal (VDS) paradigm adapted to a touchscreen environment was used to assess impulsivity and attention in Sprague-Dawley rats at 2, 3, 5, 8, and 14 months of age. After one week of training, animals were tested at different ages in 3-stage testing protocol. Additionally, prior to testing, animals were fed a restricted diet (16 g/animal). The first stage included 20 trials with inter-trial interval of 6s (ITI6si) that reflected motor impulsivity. The second stage, with 60 randomly distributed trials of ITI9s or 15s, was related to delay intolerance, while the final stage (ITI6sf), similar to the first, was related to reflection impulsivity. The strict 3-day restriction diet (24h food deprivation followed by 10g/day/animal and 8g/day/animal) was applied to 14-month-old animals before testing. Gabra5 expression in the hippocampus was determined by qPCR. Results were analyzed by one-way ANOVA with or without repeated measures, followed by Sidak post-hoc test for impulsivity and attention parameters and by t-test for PCR parameters. Results: Animals aged 8 and 14 months had reduced motor impulsivity (p<0.01 for both groups) and delay intolerance (p<0.05 for both groups) and higher number of omissions (p<0.05 for both groups) compared to animals aged 2, 3 and 5 months of age. In addition, half of the animals were unable to successfully complete a task after 14 months. After rigorous food restriction in 14-month-old animals, the level of impulsivity (ITI9s and ITI15s) and attention (number of omissions) returned to the control level (2 and 3 months of age) compared to the performance of 14-month-old animals prior to rigorous food restriction (p<0.05). Further, the peak of reflection impulsivity (ITI6sf) was reached at 5 months compared to all other groups (p<0.01). No changes in Gabra5 expression in hippocampus were detected in 14-month-old compared to 3-month-old animals. Conclusion: From 8 months of age onwards, rats showed reduced impulsivity in the VDS stages where motor impulsivity and delay intolerance were tested, followed by attention deficits. After strict food restriction in 14-month-old animals, delay intolerance and attention were restored, suggesting the prominent role of motivation in controlling these processes, independently of Gabra5 expression levels in the hippocampus. Since the VDS paradigm aims to assess reward-related impulsivity based on cognition and motivation, it is suspected that results related to impaired cognition in older animals in other cognitive tests should be interpreted with caution, and with additional observation of motivation.",
publisher = "European College of Neuropsychopharmacology (ECNP)",
journal = "36th ECPN congress, 7th -10th October 2023, Barcelona, Spain",
title = "Reward-related impulsivity as a possible surrogate marker of motivation in aging Sprague-Dawley rats",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5520"
}

Aranđelović, J., Kojić, J., Mirković, K., Jančić, I., Todorović, L.,& Savić, M.. (2023). Reward-related impulsivity as a possible surrogate marker of motivation in aging Sprague-Dawley rats. in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain
European College of Neuropsychopharmacology (ECNP)..
https://hdl.handle.net/21.15107/rcub_farfar_5520

Aranđelović J, Kojić J, Mirković K, Jančić I, Todorović L, Savić M. Reward-related impulsivity as a possible surrogate marker of motivation in aging Sprague-Dawley rats. in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5520 .

Aranđelović, Jovana, Kojić, Jana, Mirković, Kristina, Jančić, Ivan, Todorović, Lidija, Savić, Miroslav, "Reward-related impulsivity as a possible surrogate marker of motivation in aging Sprague-Dawley rats" in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5520 .

TY  - CONF
AU  - Mitrović, Jelena
AU  - Ilić, Tanja
AU  - Jančić, Ivan
AU  - Bufan, Biljana
AU  - Savić, Miroslav
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5090
AB  - Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration
Jelena Mitrović1, Tanja Ilić1, Ivan Jančić2, Biljana Bufan2, Miroslav Savić3, Snežana Savić1
1 Department of Pharmaceutical Technology and Cosmetology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
2 Department of Microbiology and Immunology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
3 Department of Pharmacology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
Estimation of endotoxin level in parenteral formulations is a prerequisite for numerous in vitro tests in preclinical studies and for future clinical development. However, the Limulus amoebocyte lysate (LAL) test in formulations containing nanoparticles could often lead to misinterpretation of results. Therefore, we tested if endotoxins could be detected in nanocrystal dispersions by the commercial gel clot assay kit. Nanocrystals of DK-I-56-1 (7‑methoxy‑2-(4‑methoxy‑d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one) were prepared by wet-ball milling, lyophilized and reconstituted with water for injection prior experiment. Different dilutions of nanocrystal dispersion in LAL reagent water were prepared as well as positive and negative control. Despite difficulties to detect gel clots, they were visible in the sample at dilutions 1:75 and below. According to the protocol, the endotoxin limit was estimated to be 25.00 EU/ml, which corresponds to <12.50 EU/mg of DK-I-56-1. This value relates to the endotoxin limit for diazepam, with the similar dosing regimen as proposed for DK-I-56-1.
C3  - NANOGVA Symposium, October 5th - 6th, 2023, Geneva, Switzerland
T1  - Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5090
ER  - 

@conference{
author = "Mitrović, Jelena and Ilić, Tanja and Jančić, Ivan and Bufan, Biljana and Savić, Miroslav and Savić, Snežana",
year = "2023",
abstract = "Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration
Jelena Mitrović1, Tanja Ilić1, Ivan Jančić2, Biljana Bufan2, Miroslav Savić3, Snežana Savić1
1 Department of Pharmaceutical Technology and Cosmetology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
2 Department of Microbiology and Immunology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
3 Department of Pharmacology, University of Belgrade – Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, Serbia
Estimation of endotoxin level in parenteral formulations is a prerequisite for numerous in vitro tests in preclinical studies and for future clinical development. However, the Limulus amoebocyte lysate (LAL) test in formulations containing nanoparticles could often lead to misinterpretation of results. Therefore, we tested if endotoxins could be detected in nanocrystal dispersions by the commercial gel clot assay kit. Nanocrystals of DK-I-56-1 (7‑methoxy‑2-(4‑methoxy‑d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one) were prepared by wet-ball milling, lyophilized and reconstituted with water for injection prior experiment. Different dilutions of nanocrystal dispersion in LAL reagent water were prepared as well as positive and negative control. Despite difficulties to detect gel clots, they were visible in the sample at dilutions 1:75 and below. According to the protocol, the endotoxin limit was estimated to be 25.00 EU/ml, which corresponds to <12.50 EU/mg of DK-I-56-1. This value relates to the endotoxin limit for diazepam, with the similar dosing regimen as proposed for DK-I-56-1.",
journal = "NANOGVA Symposium, October 5th - 6th, 2023, Geneva, Switzerland",
title = "Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5090"
}

Mitrović, J., Ilić, T., Jančić, I., Bufan, B., Savić, M.,& Savić, S.. (2023). Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration. in NANOGVA Symposium, October 5th - 6th, 2023, Geneva, Switzerland.
https://hdl.handle.net/21.15107/rcub_farfar_5090

Mitrović J, Ilić T, Jančić I, Bufan B, Savić M, Savić S. Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration. in NANOGVA Symposium, October 5th - 6th, 2023, Geneva, Switzerland. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5090 .

Mitrović, Jelena, Ilić, Tanja, Jančić, Ivan, Bufan, Biljana, Savić, Miroslav, Savić, Snežana, "Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration" in NANOGVA Symposium, October 5th - 6th, 2023, Geneva, Switzerland (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5090 .

TY  - CONF
AU  - Mirković, Kristina
AU  - Aranđelović, Jovana
AU  - Kojić, Jana
AU  - Stevanović, Vladimir
AU  - Batinić, Bojan
AU  - Todorović, Vanja
AU  - Đoković, Jelena
AU  - Santrač, Anja
AU  - Major, Tamara
AU  - Savić, Miroslav
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5521
AB  - Introduction: Ciprofloxacin is a fluoroquinolone antibiotic commonly used to treat various bacterial infections, with a potential to induce adverse mood effects in patients. Since the molecular mechanism of ciprofloxacin-induced neurotoxicity is poorly understood, we aimed to identify behavioral changes and corresponding neurotransmitter pattern after its prolonged administration in rats. We screened for untoward effects of ciprofloxacin on locomotor activity, despair, anhedonia, object recognition memory, and anxiety, as behavioral domains affected in various psychiatric diseases. Methodology: Three-month old male Sprague-Dawley rats were orally gavaged with ciprofloxacin (20 or 100 mg/kg) or solvent (0.5% methyl cellulose solution) each day for 4 weeks (n=80). One group of animals (n=40) passed the open field (OF), novel object recognition test (NORT), and forced swimming test (FST). Another group (n=40) underwent elevated plus maze (EPM) and sucrose preference test (SPT). After the completion of behavioral battery, the prefrontal cortex and cerebrospinal fluid (CSF) were collected. The neurotransmitters and metabolites of the kynurenine pathway were determined in the prefrontal cortex (PFC) through HPLC-MS/MS. Additionally, levels of interleukin-2 (IL-2) in CSF were quantified with Luminex. Behavioral and molecular parameters were analyzed by one-way ANOVA followed by Dunnett post hoc test in GraphPad Prism 9. Results: In FST, the treatment with high dose of ciprofloxacin decreased the swim time compared to control, which could be related to induction of despair-like behavior (p<0.05). The ciprofloxacin treatment did not affect object memory in NORT. In OF, the distance travelled and the number of rotations were not changed after treatment with ciprofloxacin compared to the control group. Further, animals treated with ciprofloxacin did not show changes in parameters in EPM and SPT. The acetylcholine levels in PFC were increased after ciprofloxacin treatment (p<0.05) in comparison with controls, which could be associated with depressed mood states. In line with that, high dose of ciprofloxacin treatment showed the tendency to decrease and increase levels of GABA and dopamine, respectively, but without reaching the statistical significance (p=0.07 and p=0.06). No changes in kynurenine pathway were observed after the treatment. The IL-2 concentration in CSF was increased after prolonged administration of low dose of ciprofloxacin treatment compared to the control levels (p<0.05), which could imply immunological stimulation of T lymphocytes and potential neuroinflammation. Conclusion: The despair behavior after treatment with high dose of ciprofloxacin was accompanied by increased levels of acetylcholine in PFC. Furthermore, the high dose of ciprofloxacin treatment showed tendency to decrease GABA levels, and increase dopamine levels in PFC, which could be connected to psychiatric adverse effects. Nonetheless, further studies are essential to confirm these neurotransmitter changes. On the other hand, the low dose of ciprofloxacin treatment elicited the increase of IL-2, which could be a marker of neuroinflammation-related neurotoxicity. In the future, efforts should be made to examine the role of IL-2 in the interaction of the immune system and the central nervous system, as its potential significance as a biomarker.
PB  - European College of Neuropsychopharmacology (ECNP)
C3  - 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain
T1  - Deciphering ciprofloxacin-induced neurotoxicity: behavioral and molecular profiling of ciprofloxacin treatment in rats
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5521
ER  - 

@conference{
author = "Mirković, Kristina and Aranđelović, Jovana and Kojić, Jana and Stevanović, Vladimir and Batinić, Bojan and Todorović, Vanja and Đoković, Jelena and Santrač, Anja and Major, Tamara and Savić, Miroslav",
year = "2023",
abstract = "Introduction: Ciprofloxacin is a fluoroquinolone antibiotic commonly used to treat various bacterial infections, with a potential to induce adverse mood effects in patients. Since the molecular mechanism of ciprofloxacin-induced neurotoxicity is poorly understood, we aimed to identify behavioral changes and corresponding neurotransmitter pattern after its prolonged administration in rats. We screened for untoward effects of ciprofloxacin on locomotor activity, despair, anhedonia, object recognition memory, and anxiety, as behavioral domains affected in various psychiatric diseases. Methodology: Three-month old male Sprague-Dawley rats were orally gavaged with ciprofloxacin (20 or 100 mg/kg) or solvent (0.5% methyl cellulose solution) each day for 4 weeks (n=80). One group of animals (n=40) passed the open field (OF), novel object recognition test (NORT), and forced swimming test (FST). Another group (n=40) underwent elevated plus maze (EPM) and sucrose preference test (SPT). After the completion of behavioral battery, the prefrontal cortex and cerebrospinal fluid (CSF) were collected. The neurotransmitters and metabolites of the kynurenine pathway were determined in the prefrontal cortex (PFC) through HPLC-MS/MS. Additionally, levels of interleukin-2 (IL-2) in CSF were quantified with Luminex. Behavioral and molecular parameters were analyzed by one-way ANOVA followed by Dunnett post hoc test in GraphPad Prism 9. Results: In FST, the treatment with high dose of ciprofloxacin decreased the swim time compared to control, which could be related to induction of despair-like behavior (p<0.05). The ciprofloxacin treatment did not affect object memory in NORT. In OF, the distance travelled and the number of rotations were not changed after treatment with ciprofloxacin compared to the control group. Further, animals treated with ciprofloxacin did not show changes in parameters in EPM and SPT. The acetylcholine levels in PFC were increased after ciprofloxacin treatment (p<0.05) in comparison with controls, which could be associated with depressed mood states. In line with that, high dose of ciprofloxacin treatment showed the tendency to decrease and increase levels of GABA and dopamine, respectively, but without reaching the statistical significance (p=0.07 and p=0.06). No changes in kynurenine pathway were observed after the treatment. The IL-2 concentration in CSF was increased after prolonged administration of low dose of ciprofloxacin treatment compared to the control levels (p<0.05), which could imply immunological stimulation of T lymphocytes and potential neuroinflammation. Conclusion: The despair behavior after treatment with high dose of ciprofloxacin was accompanied by increased levels of acetylcholine in PFC. Furthermore, the high dose of ciprofloxacin treatment showed tendency to decrease GABA levels, and increase dopamine levels in PFC, which could be connected to psychiatric adverse effects. Nonetheless, further studies are essential to confirm these neurotransmitter changes. On the other hand, the low dose of ciprofloxacin treatment elicited the increase of IL-2, which could be a marker of neuroinflammation-related neurotoxicity. In the future, efforts should be made to examine the role of IL-2 in the interaction of the immune system and the central nervous system, as its potential significance as a biomarker.",
publisher = "European College of Neuropsychopharmacology (ECNP)",
journal = "36th ECPN congress, 7th -10th October 2023, Barcelona, Spain",
title = "Deciphering ciprofloxacin-induced neurotoxicity: behavioral and molecular profiling of ciprofloxacin treatment in rats",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5521"
}

Mirković, K., Aranđelović, J., Kojić, J., Stevanović, V., Batinić, B., Todorović, V., Đoković, J., Santrač, A., Major, T.,& Savić, M.. (2023). Deciphering ciprofloxacin-induced neurotoxicity: behavioral and molecular profiling of ciprofloxacin treatment in rats. in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain
European College of Neuropsychopharmacology (ECNP)..
https://hdl.handle.net/21.15107/rcub_farfar_5521

Mirković K, Aranđelović J, Kojić J, Stevanović V, Batinić B, Todorović V, Đoković J, Santrač A, Major T, Savić M. Deciphering ciprofloxacin-induced neurotoxicity: behavioral and molecular profiling of ciprofloxacin treatment in rats. in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5521 .

Mirković, Kristina, Aranđelović, Jovana, Kojić, Jana, Stevanović, Vladimir, Batinić, Bojan, Todorović, Vanja, Đoković, Jelena, Santrač, Anja, Major, Tamara, Savić, Miroslav, "Deciphering ciprofloxacin-induced neurotoxicity: behavioral and molecular profiling of ciprofloxacin treatment in rats" in 36th ECPN congress, 7th -10th October 2023, Barcelona, Spain (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5521 .

TY  - CONF
AU  - Živančević, Katarina
AU  - Baralić, Katarina
AU  - Vukelić, Dragana
AU  - Marić, Đurđica
AU  - Kotur-Stevuljević, Jelena
AU  - Ivanišević, Jasmina
AU  - Savić, Miroslav
AU  - Batinić, Bojan
AU  - Janković, Radmila
AU  - Buha-Đorđević, Aleksandra
AU  - Ćurčić, Marijana
AU  - Bulat, Zorica
AU  - Antonijević, Biljana
AU  - Đukić-Ćosić, Danijela
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5424
AB  - Izloženost olovu (Pb), kadmijumu (Cd), živi (Hg) i arsenu (As) iz životne sredine, kako pojedinačno tako
i njihovim smešama jedan je od faktora za razvoj brojnih zdravstvenih poremećaja, među kojima su i
štetni efekti na nivou centralnog nervnog sistema. ...
AB  - Lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As) are top ranked toxic metal(oid)s on the list of
the most important environmental pollutants from the aspect of public health. Literature data show
that exposure to individual metals, as well as their mixtures, is one of the factors for numerous health
disorders development, including adverse effects on the central nervous system. ...
PB  - Udruženje toksikologa Srbije
C3  - 13th international congress of the Serbian society of toxicology & 1st toxSEE regional conference, 10-12 May, 2023, Belgrade, Abstract Book
T1  - Neurotoksičnost smeše niskih doza olova, kadmijuma, žive i arsena značajnih za izloženost iz životne sredine
T1  - Neurotoxicity of lead, cadmium, mercury and arsenic low dosed mixtures relevant to environmental exposure
SP  - 37
EP  - 38
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5424
ER  - 

@conference{
author = "Živančević, Katarina and Baralić, Katarina and Vukelić, Dragana and Marić, Đurđica and Kotur-Stevuljević, Jelena and Ivanišević, Jasmina and Savić, Miroslav and Batinić, Bojan and Janković, Radmila and Buha-Đorđević, Aleksandra and Ćurčić, Marijana and Bulat, Zorica and Antonijević, Biljana and Đukić-Ćosić, Danijela",
year = "2023",
abstract = "Izloženost olovu (Pb), kadmijumu (Cd), živi (Hg) i arsenu (As) iz životne sredine, kako pojedinačno tako
i njihovim smešama jedan je od faktora za razvoj brojnih zdravstvenih poremećaja, među kojima su i
štetni efekti na nivou centralnog nervnog sistema. ..., Lead (Pb), cadmium (Cd), mercury (Hg) and arsenic (As) are top ranked toxic metal(oid)s on the list of
the most important environmental pollutants from the aspect of public health. Literature data show
that exposure to individual metals, as well as their mixtures, is one of the factors for numerous health
disorders development, including adverse effects on the central nervous system. ...",
publisher = "Udruženje toksikologa Srbije",
journal = "13th international congress of the Serbian society of toxicology & 1st toxSEE regional conference, 10-12 May, 2023, Belgrade, Abstract Book",
title = "Neurotoksičnost smeše niskih doza olova, kadmijuma, žive i arsena značajnih za izloženost iz životne sredine, Neurotoxicity of lead, cadmium, mercury and arsenic low dosed mixtures relevant to environmental exposure",
pages = "37-38",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5424"
}

Živančević, K., Baralić, K., Vukelić, D., Marić, Đ., Kotur-Stevuljević, J., Ivanišević, J., Savić, M., Batinić, B., Janković, R., Buha-Đorđević, A., Ćurčić, M., Bulat, Z., Antonijević, B.,& Đukić-Ćosić, D.. (2023). Neurotoksičnost smeše niskih doza olova, kadmijuma, žive i arsena značajnih za izloženost iz životne sredine. in 13th international congress of the Serbian society of toxicology & 1st toxSEE regional conference, 10-12 May, 2023, Belgrade, Abstract Book
Udruženje toksikologa Srbije., 37-38.
https://hdl.handle.net/21.15107/rcub_farfar_5424

Živančević K, Baralić K, Vukelić D, Marić Đ, Kotur-Stevuljević J, Ivanišević J, Savić M, Batinić B, Janković R, Buha-Đorđević A, Ćurčić M, Bulat Z, Antonijević B, Đukić-Ćosić D. Neurotoksičnost smeše niskih doza olova, kadmijuma, žive i arsena značajnih za izloženost iz životne sredine. in 13th international congress of the Serbian society of toxicology & 1st toxSEE regional conference, 10-12 May, 2023, Belgrade, Abstract Book. 2023;:37-38.
https://hdl.handle.net/21.15107/rcub_farfar_5424 .

Živančević, Katarina, Baralić, Katarina, Vukelić, Dragana, Marić, Đurđica, Kotur-Stevuljević, Jelena, Ivanišević, Jasmina, Savić, Miroslav, Batinić, Bojan, Janković, Radmila, Buha-Đorđević, Aleksandra, Ćurčić, Marijana, Bulat, Zorica, Antonijević, Biljana, Đukić-Ćosić, Danijela, "Neurotoksičnost smeše niskih doza olova, kadmijuma, žive i arsena značajnih za izloženost iz životne sredine" in 13th international congress of the Serbian society of toxicology & 1st toxSEE regional conference, 10-12 May, 2023, Belgrade, Abstract Book (2023):37-38,
https://hdl.handle.net/21.15107/rcub_farfar_5424 .

TY  - CONF
AU  - Savić, Miroslav
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5116
AB  - Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs
Miroslav Savić
Department of Pharmacology, Faculty of Pharmacy, University of Belgrade, Serbia
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 IACP conference, World Conferences on Physico Chemical Methods in Drug Discovery and Development and on ADMET and DMPK; September 4th - 6th, Belgrade, Serbia
T1  - Neuropharmacokinetics: the secret life of - old and novel - psychopharmacological drugs
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5116
ER  - 

@conference{
author = "Savić, Miroslav",
year = "2023",
abstract = "Neuro pharmacokinetics: the secret life of - old and novel - psychopharmacological drugs
Miroslav Savić
Department of Pharmacology, Faculty of Pharmacy, University of Belgrade, Serbia
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 IACP conference, World Conferences on Physico Chemical Methods in Drug Discovery and Development and on ADMET and DMPK; September 4th - 6th, Belgrade, Serbia",
title = "Neuropharmacokinetics: the secret life of - old and novel - psychopharmacological drugs",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5116"
}

Savić, M.. (2023). Neuropharmacokinetics: the secret life of - old and novel - psychopharmacological drugs. in 10th IACP conference, World Conferences on Physico Chemical Methods in Drug Discovery and Development and on ADMET and DMPK; September 4th - 6th, Belgrade, Serbia
International Association of Physical Chemists..
https://hdl.handle.net/21.15107/rcub_farfar_5116

Savić M. Neuropharmacokinetics: the secret life of - old and novel - psychopharmacological drugs. in 10th IACP conference, World Conferences on Physico Chemical Methods in Drug Discovery and Development and on ADMET and DMPK; September 4th - 6th, Belgrade, Serbia. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5116 .

Savić, Miroslav, "Neuropharmacokinetics: the secret life of - old and novel - psychopharmacological drugs" in 10th IACP conference, World Conferences on Physico Chemical Methods in Drug Discovery and Development and on ADMET and DMPK; September 4th - 6th, Belgrade, Serbia (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5116 .

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  - 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 . .