Savić, Snežana

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Authority KeyName Variants
orcid::0000-0002-6236-9730
  • Savić, Snežana (202)
Projects
Development of micro- and nanosystems as carriers for drugs with anti-inflammatory effect and methods for their characterization Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200161 (University of Belgrade, Faculty of Pharmacy)
NanoCellEmoCog - 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 Functional physiologically active plant materials with additional values for application in pharmaceutical and food industry
Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200026 (University of Belgrade, Institute of Chemistry, Technology and Metallurgy - IChTM) Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200133 (Univeristy of Niš, Faculty of Technology, Leskovac)
Razvoj i karakterizacija koloidnih nosača za antiinflamatorne lekove German Academic Exchange Service (DAAD)
Federal Republic of Germany Development of molecules with antiinflammatory and cardioprotective activity: structural modifications, modelling, physicochemical characterization and formulation investigations
Behavioral ?ffects following repeated administration of newly synthesized ligands selective for distinct subtypes of GABAA receptor benzodiazepine binding site: comparison with standard psychopharmacologic drugs Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200146 (University of Belgrade, Faculty of Physical Chemistry)
Proučavanje sinteze, strukture i aktivnosti organskih jedinjenja prirodnog i sintetskog porekla CEEPUS project CIII-RS-1113-02-1819-M-118533 - Central European Knowledge Alliance for Teaching, Learning & Research in Pharmaceutical Technology (CEKA PharmTech).
China Scholarship Council COST Action CA16231 European Network of Vaccine Adjuvants (ENOVA)
Croatian Science Foundation, grant number IP-2019-04-6048 Development of Micro- and Nanosystems as Carriers for Drugswith Anti-inflammatory Effect and Methods for Their Characterizationand OI 175011 Biological response modifiers in physiological and path-ological conditions.
Division of Chemistry through grant CHE-1625735 to JC. The APC was funded by the Ministry of Education, Science and Technological, Development, the Republic of Serbia European Cooperation in Science and Technology (COST) Action CA16231 ENOVA
European Regional Development Fund European Social Fund
German Academic Exchange Service (DAAD) 57334785 German Academic Exchange Service (DAAD) A/11/84475
Industrial Collective Research, German Federation of Industrial Research Associations (Forschungsvereinigung der Arzneimittelhersteller) and German Federal Ministry for economic affairs and energy NeuroDeRisk - Neurotoxicity De-Risking in Preclinical Drug Discovery
STRENTEX - ERA Chair for emerging technologies and innovative research in Stretchable and Textile Electronics info:eu-repo/grantAgreement/MESTD-2020/inst/200026/RS//
Oxide-based environmentally-friendly porous materials for genotoxic substances removal Electroconducting and redox-active polymers and oligomers: synthesis, structure, properties and applications

Author's Bibliography

Textile-based wearable device for detection of date rape drugs in drinks

Stojanović, Goran M; Milić, Lazar; Endro, Ali A; Simić, Mitar; Nikolić, Ines; Savić, Miroslav; Savić, Snežana

(SAGE Publications Ltd, 2023)

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 . .
1

Injectable products' bioequivalence assessment

Krajišnik, Danina; Savić, Sanela; Ilić, Tanja; Savić, Snežana

(Nova Science Publishers, Inc., 2023)


                                            

                                            
Krajišnik, D., Savić, S., Ilić, T.,& Savić, S.. (2023). Injectable products' bioequivalence assessment. in Time-Proof Perspectives on Bioequivalence
Nova Science Publishers, Inc.., 221-251.
https://hdl.handle.net/21.15107/rcub_farfar_5534
Krajišnik D, Savić S, Ilić T, Savić S. Injectable products' bioequivalence assessment. in Time-Proof Perspectives on Bioequivalence. 2023;:221-251.
https://hdl.handle.net/21.15107/rcub_farfar_5534 .
Krajišnik, Danina, Savić, Sanela, Ilić, Tanja, Savić, Snežana, "Injectable products' bioequivalence assessment" in Time-Proof Perspectives on Bioequivalence (2023):221-251,
https://hdl.handle.net/21.15107/rcub_farfar_5534 .

Sizing up the nanomedicines: a thorough examination of techniques and data interpretation

Nikolić, Ines; Đoković, Jelena; Mehn, Dora; Guerrini, Giuditta; Colpo, Pascal; Jordan, Olivier; Savić, Snežana; Borchard, Gerrit

(French Society for Nanomedicine (SFNano), 2023)

TY  - CONF
AU  - Nikolić, Ines
AU  - Đoković, Jelena
AU  - Mehn, Dora
AU  - Guerrini, Giuditta
AU  - Colpo, Pascal
AU  - Jordan, Olivier
AU  - Savić, Snežana
AU  - Borchard, Gerrit
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5427
AB  - Size is a universal and first in mind property relevant for nanomedicine/nanomaterial
characterization, representing the most suitable measurand. However, due to the acknowledged
lack of standardized protocols that are able to provide reliable results, especially in the
biorelevant context, researchers often engage with the procedures that are successful in
acquiring data, but not necessarily the correct ones. ...
PB  - French Society for Nanomedicine (SFNano)
C3  - 9th SFNano annual meeting; December 4th - 6th, Montpellier, France
T1  - Sizing up the nanomedicines: a thorough examination of techniques and data interpretation
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5427
ER  - 
@conference{
author = "Nikolić, Ines and Đoković, Jelena and Mehn, Dora and Guerrini, Giuditta and Colpo, Pascal and Jordan, Olivier and Savić, Snežana and Borchard, Gerrit",
year = "2023",
abstract = "Size is a universal and first in mind property relevant for nanomedicine/nanomaterial
characterization, representing the most suitable measurand. However, due to the acknowledged
lack of standardized protocols that are able to provide reliable results, especially in the
biorelevant context, researchers often engage with the procedures that are successful in
acquiring data, but not necessarily the correct ones. ...",
publisher = "French Society for Nanomedicine (SFNano)",
journal = "9th SFNano annual meeting; December 4th - 6th, Montpellier, France",
title = "Sizing up the nanomedicines: a thorough examination of techniques and data interpretation",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5427"
}
Nikolić, I., Đoković, J., Mehn, D., Guerrini, G., Colpo, P., Jordan, O., Savić, S.,& Borchard, G.. (2023). Sizing up the nanomedicines: a thorough examination of techniques and data interpretation. in 9th SFNano annual meeting; December 4th - 6th, Montpellier, France
French Society for Nanomedicine (SFNano)..
https://hdl.handle.net/21.15107/rcub_farfar_5427
Nikolić I, Đoković J, Mehn D, Guerrini G, Colpo P, Jordan O, Savić S, Borchard G. Sizing up the nanomedicines: a thorough examination of techniques and data interpretation. in 9th SFNano annual meeting; December 4th - 6th, Montpellier, France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5427 .
Nikolić, Ines, Đoković, Jelena, Mehn, Dora, Guerrini, Giuditta, Colpo, Pascal, Jordan, Olivier, Savić, Snežana, Borchard, Gerrit, "Sizing up the nanomedicines: a thorough examination of techniques and data interpretation" in 9th SFNano annual meeting; December 4th - 6th, Montpellier, France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5427 .

Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances

Stanković, Tijana; Ilić, Tanja; Dobričić, Vladimir; Tošić, Anđela; Pantelić, Ivana; Savić, Snežana

(Savez farmaceutskih udruženja Srbije, 2023)

TY  - JOUR
AU  - Stanković, Tijana
AU  - Ilić, Tanja
AU  - Dobričić, Vladimir
AU  - Tošić, Anđela
AU  - Pantelić, Ivana
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5386
AB  - In order to improve the delivery of topical corticosteroids into inflammatory skin lesions
while reducing the likelihood of adverse effects, lipid nanocarriers have received increasing
attention. Hence, this study aimed to develop biocompatible nanoemulsions (NEs) and
nanostructured lipid carriers (NLCs) as carriers for fluocinolone acetonide (FA) by carefully
optimizing the formulation and process parameters. After an analysis of the relevant
physicochemical parameters and stability testing, in vitro release and permeation tests were
performed to evaluate whether the nanocarriers affected the penetration of FA into/through the
skin compared to a conventional reference product (Sinoderm® cream). The developed NEs
exhibited satisfactory physicochemical properties (droplet size <200 nm, PDI<0.2, ZP>ǀ-30ǀ mV,
pH ~ 4.75) and long-term stability. Although the developed NLCs initially had satisfactory
properties, gelation was observed within 3 months of storage, implying that further formulation
testing is required to resolve the limited stability of these systems. In vitro release/permeation
findings suggest that the developed nanocarriers (especially NEs) provide better delivery of FA
into/though the skin compared to the Sinoderm® cream. Therefore, a lecithin-based NE with a
10% lipid phase (medium-chain triglycerides/oleic acid 3:1) is a promising strategy for improved
delivery of FA to the inflamed skin, allowing for ease of application, especially to larger skin
surfaces and hairy regions.
AB  - Kako bi se poboljšala topikalna isporuka kortikosteroida u inflamatorne lezije kože i
istovremeno smanjila učestalost neželjenih efekata, posebna pažnja je usmerena ka razvoju
lipidnih nanonosača. Stoga, cilj ovog rada je bio razvoj biokompatibilnih nanoemulzija (NEs) i
nanostrukturiranih lipidnih nosača (NLCs) kao nosača za fluocinolonacetonid (FA) pažljivom
optimizacijom formulacionih i procesnih parametara. Nakon analize relevantnih fizičko-
hemijskih parametara i studije stabilnosti, in vitro ispitivanje oslobađanja i permeacije je
sprovedeno kako bi se dobio uvid u to da li razvijeni nanonosači utiču na penetraciju FA u/kroz
kožu, u poređenju sa konvencionalnim referentnim preparatom (Sinoderm® krem). Uspešno su
razvijene NEs zadovoljavajućih fizičko-hemijskih osobina (veličina kapi<200 nm, PDI<0,2,
ZP>ǀ-30ǀ mV, pH~4,75) i dugoročne stabilnosti. Iako su inicijalno posedovali zadovoljavajuće
karakteristike, NLCs su gelirali tokom tri meseca čuvanja, što ukazuje na potrebu za daljim radom
na razvoju formulacije, u cilju rešavanja problema ograničene stabilnosti ovih sistema. Nalazi in
vitro ispitivanja oslobađanja/permeacije upućuju na činjenicu da razvijeni lipidni nanonosači
(prevashodno NEs) obezbeđuju bolju isporuku FA u/kroz kožu u poređenju sa Sinoderm®
kremom. Nanoemulzije bazirane na lecitinu sa 10% uljane faze (smeša triglicerida srednje dužine
lanaca i oleinske kiseline 3:1) predstavljaju obećavajuću strategiju za poboljšanu isporuku FA u
inflamatorne promene na koži, omogućavajući laku primenu, posebno na većim površinama i
kosmatim delovima tela.
PB  - Savez farmaceutskih udruženja Srbije
T2  - Arhiv za farmaciju
T1  - Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances
T1  - Biokompatibilni lipidni nanonosači za poboljšanu isporuku fluocinolonacetonida u kožu: fizičko- hemijske osobine i in vitro učinak
VL  - 73
IS  - 5
SP  - 423
EP  - 439
DO  - 10.5937/arhfarm73-46312
ER  - 
@article{
author = "Stanković, Tijana and Ilić, Tanja and Dobričić, Vladimir and Tošić, Anđela and Pantelić, Ivana and Savić, Snežana",
year = "2023",
abstract = "In order to improve the delivery of topical corticosteroids into inflammatory skin lesions
while reducing the likelihood of adverse effects, lipid nanocarriers have received increasing
attention. Hence, this study aimed to develop biocompatible nanoemulsions (NEs) and
nanostructured lipid carriers (NLCs) as carriers for fluocinolone acetonide (FA) by carefully
optimizing the formulation and process parameters. After an analysis of the relevant
physicochemical parameters and stability testing, in vitro release and permeation tests were
performed to evaluate whether the nanocarriers affected the penetration of FA into/through the
skin compared to a conventional reference product (Sinoderm® cream). The developed NEs
exhibited satisfactory physicochemical properties (droplet size <200 nm, PDI<0.2, ZP>ǀ-30ǀ mV,
pH ~ 4.75) and long-term stability. Although the developed NLCs initially had satisfactory
properties, gelation was observed within 3 months of storage, implying that further formulation
testing is required to resolve the limited stability of these systems. In vitro release/permeation
findings suggest that the developed nanocarriers (especially NEs) provide better delivery of FA
into/though the skin compared to the Sinoderm® cream. Therefore, a lecithin-based NE with a
10% lipid phase (medium-chain triglycerides/oleic acid 3:1) is a promising strategy for improved
delivery of FA to the inflamed skin, allowing for ease of application, especially to larger skin
surfaces and hairy regions., Kako bi se poboljšala topikalna isporuka kortikosteroida u inflamatorne lezije kože i
istovremeno smanjila učestalost neželjenih efekata, posebna pažnja je usmerena ka razvoju
lipidnih nanonosača. Stoga, cilj ovog rada je bio razvoj biokompatibilnih nanoemulzija (NEs) i
nanostrukturiranih lipidnih nosača (NLCs) kao nosača za fluocinolonacetonid (FA) pažljivom
optimizacijom formulacionih i procesnih parametara. Nakon analize relevantnih fizičko-
hemijskih parametara i studije stabilnosti, in vitro ispitivanje oslobađanja i permeacije je
sprovedeno kako bi se dobio uvid u to da li razvijeni nanonosači utiču na penetraciju FA u/kroz
kožu, u poređenju sa konvencionalnim referentnim preparatom (Sinoderm® krem). Uspešno su
razvijene NEs zadovoljavajućih fizičko-hemijskih osobina (veličina kapi<200 nm, PDI<0,2,
ZP>ǀ-30ǀ mV, pH~4,75) i dugoročne stabilnosti. Iako su inicijalno posedovali zadovoljavajuće
karakteristike, NLCs su gelirali tokom tri meseca čuvanja, što ukazuje na potrebu za daljim radom
na razvoju formulacije, u cilju rešavanja problema ograničene stabilnosti ovih sistema. Nalazi in
vitro ispitivanja oslobađanja/permeacije upućuju na činjenicu da razvijeni lipidni nanonosači
(prevashodno NEs) obezbeđuju bolju isporuku FA u/kroz kožu u poređenju sa Sinoderm®
kremom. Nanoemulzije bazirane na lecitinu sa 10% uljane faze (smeša triglicerida srednje dužine
lanaca i oleinske kiseline 3:1) predstavljaju obećavajuću strategiju za poboljšanu isporuku FA u
inflamatorne promene na koži, omogućavajući laku primenu, posebno na većim površinama i
kosmatim delovima tela.",
publisher = "Savez farmaceutskih udruženja Srbije",
journal = "Arhiv za farmaciju",
title = "Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances, Biokompatibilni lipidni nanonosači za poboljšanu isporuku fluocinolonacetonida u kožu: fizičko- hemijske osobine i in vitro učinak",
volume = "73",
number = "5",
pages = "423-439",
doi = "10.5937/arhfarm73-46312"
}
Stanković, T., Ilić, T., Dobričić, V., Tošić, A., Pantelić, I.,& Savić, S.. (2023). Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije., 73(5), 423-439.
https://doi.org/10.5937/arhfarm73-46312
Stanković T, Ilić T, Dobričić V, Tošić A, Pantelić I, Savić S. Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances. in Arhiv za farmaciju. 2023;73(5):423-439.
doi:10.5937/arhfarm73-46312 .
Stanković, Tijana, Ilić, Tanja, Dobričić, Vladimir, Tošić, Anđela, Pantelić, Ivana, Savić, Snežana, "Biocompatible lipid nanocarriers for improved skin delivery of fluocinolone acetonide: physicochemical and in vitro performances" in Arhiv za farmaciju, 73, no. 5 (2023):423-439,
https://doi.org/10.5937/arhfarm73-46312 . .

Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects

Pantelić, Ivana; Ilić, Tanja; Nikolić, Ines; Savić, Snežana

(Savez farmaceutskih udruženja Srbije, 2023)

TY  - JOUR
AU  - Pantelić, Ivana
AU  - Ilić, Tanja
AU  - Nikolić, Ines
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5359
AB  - A review of recent publications reveals an increased interest in the so-called self-assembled
carriers and their applicability in drug delivery via various routes of administration. Self-assembly
denotes the process of rather spontaneous formation of ordered aggregates (sometimes under
specific conditions – e.g., pH, temperature, ionic strength), via diverse interactions. This process,
seen in many naturally occurring substances (polysaccharides, proteins, lipids), has inspired
researchers to synthetize innovative self-assembling materials or combinations of existing ones.
This paper provides a review of the recently investigated self-assembling materials and the
carriers they form, often belonging to the sphere of pharmaceutical nanotechnology. Self-
assembled carriers may provide enhanced stability, more efficient encapsulation and/or controlled
delivery of active pharmaceutical ingredients. However, the diversity of geometries obtained
(spheres, polyhedrals, ellipses, discs, porous structures, etc.) presents a significant
characterization challenge, often requiring the application of several complementary techniques
for proper evaluation of carrier size and morphology. Commonly utilized characterization
techniques for investigating physico-chemical and certain biopharmaceutical properties are
discussed, along with their advantages and disadvantages. Finally, the authors offer their critical
opinion on the outlook of self-assembled drug carriers
AB  - Pregled publikacija objavljenih poslednjih godina ukazuje na interesovanje za tzv. nosače sklone samoorganizovanju, kao i njihov potencijal za isporuku lekovitih supstanci različitim putevima primene. U ovom kontekstu, samoorganizacija označava proces relativno spontanog obrazovanja visoko uređenih agregata (koji ponekad zahteva specifične uslove -npr., pH, temperaturu, jonsku jačinu), zahvaljujući interakcijama različite prirode. Ovaj proces, karakterističan za mnoge supstance prirodnog porekla (određene polisaharide, proteine, lipide), poslužio je kao inspiracija istraživačima da osmisle i sintetišu inovativne materijale sklone samoorganizovanju, ili ispitaju kombinacije postojećih materijala. Ovaj rad pruža pregled najčešće ispitivanih materijala, odnosno nosača dobijenih samoorganizovanjem, koji često pripadaju sferi farmaceutske nanotehnologije. Nosači skloni samoorganizovanju mogu unaprediti stabilnost, efikasnost inkapsulacije i/ili kontrolisanu isporuku lekovitih supstanci. Ipak, raznolikost geometrija dobijenih nosača (sfere, poliedri, elipse, diskovi, porozne strukture, itd.) predstavlja značajan izazov za karakterizaciju, često zahtevajući primenu više komplementarnih tehnika, naročito za valjanu evaluaciju veličine i morfologije dobijenih nosača. Diskutovane su najčešće korišćene tehnike fizičko-hemijske i biofarmaceutske karakterizacije, uz isticanje njihovih prednosti i nedostataka. Na kraju, dat je kritički osvrt o izgledima za buduću primenu nosača lekovitih supstanci sklonih samoorganizovanju.
PB  - Savez farmaceutskih udruženja Srbije
T2  - Arhiv za farmaciju
T1  - Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects
T1  - Nosači lekovitih supstanci skloni samoorganizovanju - trenutni izazovi u karakterizaciji i izgledi za budućnost
VL  - 73
IS  - 5
SP  - 404
EP  - 422
DO  - 10.5937/arhfarm73-46975
ER  - 
@article{
author = "Pantelić, Ivana and Ilić, Tanja and Nikolić, Ines and Savić, Snežana",
year = "2023",
abstract = "A review of recent publications reveals an increased interest in the so-called self-assembled
carriers and their applicability in drug delivery via various routes of administration. Self-assembly
denotes the process of rather spontaneous formation of ordered aggregates (sometimes under
specific conditions – e.g., pH, temperature, ionic strength), via diverse interactions. This process,
seen in many naturally occurring substances (polysaccharides, proteins, lipids), has inspired
researchers to synthetize innovative self-assembling materials or combinations of existing ones.
This paper provides a review of the recently investigated self-assembling materials and the
carriers they form, often belonging to the sphere of pharmaceutical nanotechnology. Self-
assembled carriers may provide enhanced stability, more efficient encapsulation and/or controlled
delivery of active pharmaceutical ingredients. However, the diversity of geometries obtained
(spheres, polyhedrals, ellipses, discs, porous structures, etc.) presents a significant
characterization challenge, often requiring the application of several complementary techniques
for proper evaluation of carrier size and morphology. Commonly utilized characterization
techniques for investigating physico-chemical and certain biopharmaceutical properties are
discussed, along with their advantages and disadvantages. Finally, the authors offer their critical
opinion on the outlook of self-assembled drug carriers, Pregled publikacija objavljenih poslednjih godina ukazuje na interesovanje za tzv. nosače sklone samoorganizovanju, kao i njihov potencijal za isporuku lekovitih supstanci različitim putevima primene. U ovom kontekstu, samoorganizacija označava proces relativno spontanog obrazovanja visoko uređenih agregata (koji ponekad zahteva specifične uslove -npr., pH, temperaturu, jonsku jačinu), zahvaljujući interakcijama različite prirode. Ovaj proces, karakterističan za mnoge supstance prirodnog porekla (određene polisaharide, proteine, lipide), poslužio je kao inspiracija istraživačima da osmisle i sintetišu inovativne materijale sklone samoorganizovanju, ili ispitaju kombinacije postojećih materijala. Ovaj rad pruža pregled najčešće ispitivanih materijala, odnosno nosača dobijenih samoorganizovanjem, koji često pripadaju sferi farmaceutske nanotehnologije. Nosači skloni samoorganizovanju mogu unaprediti stabilnost, efikasnost inkapsulacije i/ili kontrolisanu isporuku lekovitih supstanci. Ipak, raznolikost geometrija dobijenih nosača (sfere, poliedri, elipse, diskovi, porozne strukture, itd.) predstavlja značajan izazov za karakterizaciju, često zahtevajući primenu više komplementarnih tehnika, naročito za valjanu evaluaciju veličine i morfologije dobijenih nosača. Diskutovane su najčešće korišćene tehnike fizičko-hemijske i biofarmaceutske karakterizacije, uz isticanje njihovih prednosti i nedostataka. Na kraju, dat je kritički osvrt o izgledima za buduću primenu nosača lekovitih supstanci sklonih samoorganizovanju.",
publisher = "Savez farmaceutskih udruženja Srbije",
journal = "Arhiv za farmaciju",
title = "Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects, Nosači lekovitih supstanci skloni samoorganizovanju - trenutni izazovi u karakterizaciji i izgledi za budućnost",
volume = "73",
number = "5",
pages = "404-422",
doi = "10.5937/arhfarm73-46975"
}
Pantelić, I., Ilić, T., Nikolić, I.,& Savić, S.. (2023). Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije., 73(5), 404-422.
https://doi.org/10.5937/arhfarm73-46975
Pantelić I, Ilić T, Nikolić I, Savić S. Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects. in Arhiv za farmaciju. 2023;73(5):404-422.
doi:10.5937/arhfarm73-46975 .
Pantelić, Ivana, Ilić, Tanja, Nikolić, Ines, Savić, Snežana, "Self-assembled carriers as drug delivery systems: current characterization challenges and future prospects" in Arhiv za farmaciju, 73, no. 5 (2023):404-422,
https://doi.org/10.5937/arhfarm73-46975 . .

The Landscape of Nanomedicines: An Expert Perspective

Nikolić, Ines; Filipić, Brankica; Marija Petrović; Jordan, Olivier; Savić, Snežana; Borchard, Gerrit

(Savez farmaceutskih udruženja Srbije, 2023)

TY  - JOUR
AU  - Nikolić, Ines
AU  - Filipić, Brankica
AU  - Marija Petrović
AU  - Jordan, Olivier
AU  - Savić, Snežana
AU  - Borchard, Gerrit
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5358
AB  - The field of nanotechnology is at the forefront of a scientific revolution, where the term
“nano” transcends mere size and opens the door to enormous possibilities.
In the context of drug development, the selection of a suitable drug delivery system
(corresponding to a certain active pharmaceutical ingredient) is a pivotal decision. Accordingly,
nanosystems have emerged as a promising avenue, offering innovative solutions, and gaining
recognition for addressing healthcare issues.
While these products hold immense promise, they have faced certain complexities in their
translation from the preclinical to the clinical setting, reflected in the lack of proper assessment
protocols for quality and safety aspects and, consequently, an insufficiently defined regulatory
environment. Since the groundbreaking US Food and Drug Administration (FDA) approval of
liposomal doxorubicin in 1995, approximately 80 nanomedicine products have received regulatory
approval so far. Recent attention has gravitated toward lipid-based nanomedicines, particularly in
the development of mRNA vaccines during the COVID-19 pandemic, further highlighting their
significance. However, the relatively modest number of approved nanomedicines compared to the
extensive research efforts raises important questions and underscores areas of uncertainty.
This article provides an overview of the challenges in defining nanomedicines, their
properties, the complexities of regulatory frameworks, and the imperative for standardized
characterization protocols.
AB  - Polje nanotehnologije se nalazi na čelu naučne revolucije, gde se termin "nano" izdiže iznad pukog označavanja veličine, otvarajući vrata novim mogućnostima. U kontekstu razvoja lekova, izbor odgovarajućeg sistema za isporuku / nosača (koji odgovara određenoj aktivnoj supstanci) predstavlja ključnu odluku. U tom kontekstu, nanosistemi već određeno vreme predstavljaju inovativna rešenja. Iako farmaceutski nanosistemi nose ogroman potencijal, suočavaju se sa određenim izazovima u pogledu translacije sa prekliničkog na klinički nivo, što se ogleda u nedostatku odgovarajućih protokola za ispitivanje kvaliteta i bezbednosti i, shodno tome, nedefinisanom regulatornom okruženju. Od revolucionarnog odobrenja liposomalnog doksorubicina od strane Američke agencije za hranu i lekove 1995. godine, pa sve do danas, oko 80 nano formulacija (nanolekova) odobreno je za kliničku primenu. Odnedavno je intenzivnija pažnja usmerena ka nanoformulacijama baziranim na lipidima, što je delom posledica razvoja mRNK vakcina tokom pandemije COVID-19. Međutim, relativno skroman nastup nanolekova na tržištu (u poređenju sa obimnim istraživačkim naporima i finansijskim ulaganjima u ovu oblast) otvara važna pitanja. Ovaj rad pruža pregled izazova u definisanju nanolekova, njihovih svojstava, kompleksnosti regulatornih okvira i imperativa za stvaranje standardizovanih protokola karakterizacije.
PB  - Savez farmaceutskih udruženja Srbije
T2  - Arhiv za farmaciju
T1  - The Landscape of Nanomedicines: An Expert Perspective
T1  - "Pejzažni" prikaz nanolekova - ekspertska perspektiva
VL  - 73
IS  - 5
SP  - 390
EP  - 403
DO  - 10.5937/arhfarm73-46686
ER  - 
@article{
author = "Nikolić, Ines and Filipić, Brankica and Marija Petrović and Jordan, Olivier and Savić, Snežana and Borchard, Gerrit",
year = "2023",
abstract = "The field of nanotechnology is at the forefront of a scientific revolution, where the term
“nano” transcends mere size and opens the door to enormous possibilities.
In the context of drug development, the selection of a suitable drug delivery system
(corresponding to a certain active pharmaceutical ingredient) is a pivotal decision. Accordingly,
nanosystems have emerged as a promising avenue, offering innovative solutions, and gaining
recognition for addressing healthcare issues.
While these products hold immense promise, they have faced certain complexities in their
translation from the preclinical to the clinical setting, reflected in the lack of proper assessment
protocols for quality and safety aspects and, consequently, an insufficiently defined regulatory
environment. Since the groundbreaking US Food and Drug Administration (FDA) approval of
liposomal doxorubicin in 1995, approximately 80 nanomedicine products have received regulatory
approval so far. Recent attention has gravitated toward lipid-based nanomedicines, particularly in
the development of mRNA vaccines during the COVID-19 pandemic, further highlighting their
significance. However, the relatively modest number of approved nanomedicines compared to the
extensive research efforts raises important questions and underscores areas of uncertainty.
This article provides an overview of the challenges in defining nanomedicines, their
properties, the complexities of regulatory frameworks, and the imperative for standardized
characterization protocols., Polje nanotehnologije se nalazi na čelu naučne revolucije, gde se termin "nano" izdiže iznad pukog označavanja veličine, otvarajući vrata novim mogućnostima. U kontekstu razvoja lekova, izbor odgovarajućeg sistema za isporuku / nosača (koji odgovara određenoj aktivnoj supstanci) predstavlja ključnu odluku. U tom kontekstu, nanosistemi već određeno vreme predstavljaju inovativna rešenja. Iako farmaceutski nanosistemi nose ogroman potencijal, suočavaju se sa određenim izazovima u pogledu translacije sa prekliničkog na klinički nivo, što se ogleda u nedostatku odgovarajućih protokola za ispitivanje kvaliteta i bezbednosti i, shodno tome, nedefinisanom regulatornom okruženju. Od revolucionarnog odobrenja liposomalnog doksorubicina od strane Američke agencije za hranu i lekove 1995. godine, pa sve do danas, oko 80 nano formulacija (nanolekova) odobreno je za kliničku primenu. Odnedavno je intenzivnija pažnja usmerena ka nanoformulacijama baziranim na lipidima, što je delom posledica razvoja mRNK vakcina tokom pandemije COVID-19. Međutim, relativno skroman nastup nanolekova na tržištu (u poređenju sa obimnim istraživačkim naporima i finansijskim ulaganjima u ovu oblast) otvara važna pitanja. Ovaj rad pruža pregled izazova u definisanju nanolekova, njihovih svojstava, kompleksnosti regulatornih okvira i imperativa za stvaranje standardizovanih protokola karakterizacije.",
publisher = "Savez farmaceutskih udruženja Srbije",
journal = "Arhiv za farmaciju",
title = "The Landscape of Nanomedicines: An Expert Perspective, "Pejzažni" prikaz nanolekova - ekspertska perspektiva",
volume = "73",
number = "5",
pages = "390-403",
doi = "10.5937/arhfarm73-46686"
}
Nikolić, I., Filipić, B., Marija Petrović, Jordan, O., Savić, S.,& Borchard, G.. (2023). The Landscape of Nanomedicines: An Expert Perspective. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije., 73(5), 390-403.
https://doi.org/10.5937/arhfarm73-46686
Nikolić I, Filipić B, Marija Petrović, Jordan O, Savić S, Borchard G. The Landscape of Nanomedicines: An Expert Perspective. in Arhiv za farmaciju. 2023;73(5):390-403.
doi:10.5937/arhfarm73-46686 .
Nikolić, Ines, Filipić, Brankica, Marija Petrović, Jordan, Olivier, Savić, Snežana, Borchard, Gerrit, "The Landscape of Nanomedicines: An Expert Perspective" in Arhiv za farmaciju, 73, no. 5 (2023):390-403,
https://doi.org/10.5937/arhfarm73-46686 . .

Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test

Cekić, Nebojša; Savić, Sanela; Savić, Snežana

(Savez farmaceutskih udruženja Srbije, 2023)

TY  - JOUR
AU  - Cekić, Nebojša
AU  - Savić, Sanela
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5357
AB  - The assessment of stability in emulsion-based topical preparations can be approached through
real-time monitoring and/or accelerated methods, drawing predictions from pertinent stability-related
physicochemical parameters. Ensuring the robustness and durability of topical products during
storage, transport, and application necessitates thorough stability testing. However, due to the diversity
of emulsion types and their intended use, there is no universal standard test, placing the liability on
formulators/manufacturer to tailor appropriate assessments. Notably, topical emulsions, particularly
cosmetic variants, often exhibit impressive stability with extended shelf lives. Nonetheless, evaluating
their stability and decision-making remain challenging and time-consuming in industrial contexts.
This underscores the demand for alternative testing protocols that expedite stability assessments and
predict emulsion-based product stability accurately. This article comprehensively surveys literature,
enriched with practical insights, exploring core mechanisms behind emulsion stability and prevention
of instability. The discussion encompasses diverse approaches to stability assessment, revealing
methodologies and parameters under examination during testing. Particular focus is placed on the
dynamic-mechanical thermoanalysis (DMTA) method explored as a rapid, rheologically-based
alternative to the conventional freeze-thaw test, emphasizing its usefulness for expediting the stability
evaluation of emulsion-based topical preparations.
AB  - Procena stabilnosti emulzionih preparata za topikalnu primenu može se sprovesti praćenjem promena u realnom vremenu i/ili primenom ubrzanih metoda, te predviđanjem stabilnosti i roka trajanja proizvoda na osnovu merenja relevantnih fizičkohemijskih parametara tokom ispitivanja. Kako bi se obezbedila robusnost i dugoročnost emulzionih proizvoda za kožu tokom čuvanja, transporta i primene, neophodno je sprovesti pažljivo isplanirano, opsežno ispitivanje stabilnosti. Međutim, imajući u vidu različite tipove emulzija i njihovu namenu, ne postoji univerzalni standardni protokol za ispitivanje stabilnosti, što formulatore/proizvođača čini odgovornim kada je u pitanju izbor odgovarajućeg testa i metodologije. Evidentno je da emulzije za topikalnu primenu, a posebno kozmetičke emulzije, često pokazuju visoku stabilnost sa dugim rokovima upotrebe. S druge strane, procena stabilnosti ovakvih emulzija i donošenje odgovarajućih odluka i dalje ostaje izazov u industrijskom okruženju i zahteva dosta vremena, što nameće potrebu za alternativnim protokolima koji omogućavaju ubrzano ispitivanje, ali i uspešno predviđanje stabilnosti emulzionih proizvoda. Prikazani rad daje sveobuhvatni pregled literature prožet praktičnim pogledima na ključne fenomene odgovorne za stabilnost emulzija, zatim daje uvid u različite pristupe za procenu njihove stabilnosti, uključujući metodologije koje se koriste i parametre koji se prate tokom ispitivanja. Rad u poseban fokus stavlja dinamičko-mehanički termoanalitički (DMTA) metod kao brzu reološku alternativu konvencionalnom testu smrzavanje-odmrzavanje, posebno ističući primenljivost metoda za ubrzano ispitivanje stabilnosti emulzionih preparata za topikalnu primenu.
PB  - Savez farmaceutskih udruženja Srbije
T2  - Arhiv za farmaciju
T1  - Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test
T1  - Procena stabilnosti emulzionih preparata za topikalnu primenu - vrednost dinamičkomehaničkog termoanalitičkog (DMTA) testa kao brze reološke alternative konvencionalnom testu smrzavanje-odmrzavanje
VL  - 73
IS  - 5
SP  - 358
EP  - 389
DO  - 10.5937/arhfarm73-46319
ER  - 
@article{
author = "Cekić, Nebojša and Savić, Sanela and Savić, Snežana",
year = "2023",
abstract = "The assessment of stability in emulsion-based topical preparations can be approached through
real-time monitoring and/or accelerated methods, drawing predictions from pertinent stability-related
physicochemical parameters. Ensuring the robustness and durability of topical products during
storage, transport, and application necessitates thorough stability testing. However, due to the diversity
of emulsion types and their intended use, there is no universal standard test, placing the liability on
formulators/manufacturer to tailor appropriate assessments. Notably, topical emulsions, particularly
cosmetic variants, often exhibit impressive stability with extended shelf lives. Nonetheless, evaluating
their stability and decision-making remain challenging and time-consuming in industrial contexts.
This underscores the demand for alternative testing protocols that expedite stability assessments and
predict emulsion-based product stability accurately. This article comprehensively surveys literature,
enriched with practical insights, exploring core mechanisms behind emulsion stability and prevention
of instability. The discussion encompasses diverse approaches to stability assessment, revealing
methodologies and parameters under examination during testing. Particular focus is placed on the
dynamic-mechanical thermoanalysis (DMTA) method explored as a rapid, rheologically-based
alternative to the conventional freeze-thaw test, emphasizing its usefulness for expediting the stability
evaluation of emulsion-based topical preparations., Procena stabilnosti emulzionih preparata za topikalnu primenu može se sprovesti praćenjem promena u realnom vremenu i/ili primenom ubrzanih metoda, te predviđanjem stabilnosti i roka trajanja proizvoda na osnovu merenja relevantnih fizičkohemijskih parametara tokom ispitivanja. Kako bi se obezbedila robusnost i dugoročnost emulzionih proizvoda za kožu tokom čuvanja, transporta i primene, neophodno je sprovesti pažljivo isplanirano, opsežno ispitivanje stabilnosti. Međutim, imajući u vidu različite tipove emulzija i njihovu namenu, ne postoji univerzalni standardni protokol za ispitivanje stabilnosti, što formulatore/proizvođača čini odgovornim kada je u pitanju izbor odgovarajućeg testa i metodologije. Evidentno je da emulzije za topikalnu primenu, a posebno kozmetičke emulzije, često pokazuju visoku stabilnost sa dugim rokovima upotrebe. S druge strane, procena stabilnosti ovakvih emulzija i donošenje odgovarajućih odluka i dalje ostaje izazov u industrijskom okruženju i zahteva dosta vremena, što nameće potrebu za alternativnim protokolima koji omogućavaju ubrzano ispitivanje, ali i uspešno predviđanje stabilnosti emulzionih proizvoda. Prikazani rad daje sveobuhvatni pregled literature prožet praktičnim pogledima na ključne fenomene odgovorne za stabilnost emulzija, zatim daje uvid u različite pristupe za procenu njihove stabilnosti, uključujući metodologije koje se koriste i parametre koji se prate tokom ispitivanja. Rad u poseban fokus stavlja dinamičko-mehanički termoanalitički (DMTA) metod kao brzu reološku alternativu konvencionalnom testu smrzavanje-odmrzavanje, posebno ističući primenljivost metoda za ubrzano ispitivanje stabilnosti emulzionih preparata za topikalnu primenu.",
publisher = "Savez farmaceutskih udruženja Srbije",
journal = "Arhiv za farmaciju",
title = "Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test, Procena stabilnosti emulzionih preparata za topikalnu primenu - vrednost dinamičkomehaničkog termoanalitičkog (DMTA) testa kao brze reološke alternative konvencionalnom testu smrzavanje-odmrzavanje",
volume = "73",
number = "5",
pages = "358-389",
doi = "10.5937/arhfarm73-46319"
}
Cekić, N., Savić, S.,& Savić, S.. (2023). Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije., 73(5), 358-389.
https://doi.org/10.5937/arhfarm73-46319
Cekić N, Savić S, Savić S. Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test. in Arhiv za farmaciju. 2023;73(5):358-389.
doi:10.5937/arhfarm73-46319 .
Cekić, Nebojša, Savić, Sanela, Savić, Snežana, "Stability evaluation of emulsion-based topical preparations: a valuable potential of dynamic- mechanical thermoanalysis (DMTA) test as a rapid rheological alternative to conventional freeze- thaw test" in Arhiv za farmaciju, 73, no. 5 (2023):358-389,
https://doi.org/10.5937/arhfarm73-46319 . .

Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety

Vukašinović, Mila; Pantelić, Ivana; Savić, Sanela; Cekić, Nebojša; Vukašinović Sekulić, Maja; Antić-Stanković, Jelena; Božić, Dragana; Tošić, Anđela; Tamburić, Slobodanka; Savić, Snežana

(MDPI, 2023)

TY  - JOUR
AU  - Vukašinović, Mila
AU  - Pantelić, Ivana
AU  - Savić, Sanela
AU  - Cekić, Nebojša
AU  - Vukašinović Sekulić, Maja
AU  - Antić-Stanković, Jelena
AU  - Božić, Dragana
AU  - Tošić, Anđela
AU  - Tamburić, Slobodanka
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5311
AB  - Bioactive peptides are promising cosmetic active ingredients that can improve skin health
and appearance. They exhibit a broad spectrum of activity, including anti-aging, antioxidant, an-
timicrobial, and anti-inflammatory effects. The aim of this study was to develop a safe, stable, and
efficacious environmentally friendly (“green”) emulsion using a milk protein hydrolysate as a model
active ingredient. Potential emulsions were formulated with biodegradable emollients, stabilized
with naturally derived mixed emulsifier, and prepared by cold process. They were evaluated for
rheological behavior (continuous rotation and oscillation tests), physical stability (dynamic me-
chanical thermal analysis—DMTA test), and texture profiles, as well as cytotoxic, antioxidant, and
antimicrobial effects. Rheological characterization revealed shear-thinning flow behavior with yield
point from continuous rotation tests and predominantly elastic character from oscillation (amplitude
and frequency sweep) tests, with small structural change detected in the DMTA test. These results
implied satisfactory rheological properties and good stability. Texture analysis revealed acceptable
spreadability and substantivity of the emulsions. The protein hydrolysate showed antioxidant activity.
The developed emulsions showed low antibacterial activity against selected microorganisms, but
this was due to the action of preservatives, not peptides. All potential emulsions showed a desirable
safety profile. The results obtained provide the basis for the next stage of formulation development,
i.e., in vivo efficacy tests.
PB  - MDPI
T2  - Cosmetics
T1  - Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety
VL  - 10
IS  - 6
DO  - 10.3390/cosmetics10060162
ER  - 
@article{
author = "Vukašinović, Mila and Pantelić, Ivana and Savić, Sanela and Cekić, Nebojša and Vukašinović Sekulić, Maja and Antić-Stanković, Jelena and Božić, Dragana and Tošić, Anđela and Tamburić, Slobodanka and Savić, Snežana",
year = "2023",
abstract = "Bioactive peptides are promising cosmetic active ingredients that can improve skin health
and appearance. They exhibit a broad spectrum of activity, including anti-aging, antioxidant, an-
timicrobial, and anti-inflammatory effects. The aim of this study was to develop a safe, stable, and
efficacious environmentally friendly (“green”) emulsion using a milk protein hydrolysate as a model
active ingredient. Potential emulsions were formulated with biodegradable emollients, stabilized
with naturally derived mixed emulsifier, and prepared by cold process. They were evaluated for
rheological behavior (continuous rotation and oscillation tests), physical stability (dynamic me-
chanical thermal analysis—DMTA test), and texture profiles, as well as cytotoxic, antioxidant, and
antimicrobial effects. Rheological characterization revealed shear-thinning flow behavior with yield
point from continuous rotation tests and predominantly elastic character from oscillation (amplitude
and frequency sweep) tests, with small structural change detected in the DMTA test. These results
implied satisfactory rheological properties and good stability. Texture analysis revealed acceptable
spreadability and substantivity of the emulsions. The protein hydrolysate showed antioxidant activity.
The developed emulsions showed low antibacterial activity against selected microorganisms, but
this was due to the action of preservatives, not peptides. All potential emulsions showed a desirable
safety profile. The results obtained provide the basis for the next stage of formulation development,
i.e., in vivo efficacy tests.",
publisher = "MDPI",
journal = "Cosmetics",
title = "Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety",
volume = "10",
number = "6",
doi = "10.3390/cosmetics10060162"
}
Vukašinović, M., Pantelić, I., Savić, S., Cekić, N., Vukašinović Sekulić, M., Antić-Stanković, J., Božić, D., Tošić, A., Tamburić, S.,& Savić, S.. (2023). Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety. in Cosmetics
MDPI., 10(6).
https://doi.org/10.3390/cosmetics10060162
Vukašinović M, Pantelić I, Savić S, Cekić N, Vukašinović Sekulić M, Antić-Stanković J, Božić D, Tošić A, Tamburić S, Savić S. Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety. in Cosmetics. 2023;10(6).
doi:10.3390/cosmetics10060162 .
Vukašinović, Mila, Pantelić, Ivana, Savić, Sanela, Cekić, Nebojša, Vukašinović Sekulić, Maja, Antić-Stanković, Jelena, Božić, Dragana, Tošić, Anđela, Tamburić, Slobodanka, Savić, Snežana, "Development of a “Green” Emulsion with a Milk Protein Hydrolysate: An Evaluation of Rheology, Texture, In Vitro Bioactivity, and Safety" in Cosmetics, 10, no. 6 (2023),
https://doi.org/10.3390/cosmetics10060162 . .
1

Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test

Gledović, Ana; Ilić, Tanja; Savić, Snežana

(APGI – “Association de Pharmacie Galénique Industrielle”, 2023)

TY  - CONF
AU  - Gledović, Ana
AU  - Ilić, Tanja
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5189
AB  - Nanoemulsions (NEs), especially those prepared by low-
energy processes, have recently been proposed as
promising carriers for natural products such as plant oils and
extracts. ...
PB  - APGI – “Association de Pharmacie Galénique Industrielle”
C3  - 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France
T1  - Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5189
ER  - 
@conference{
author = "Gledović, Ana and Ilić, Tanja and Savić, Snežana",
year = "2023",
abstract = "Nanoemulsions (NEs), especially those prepared by low-
energy processes, have recently been proposed as
promising carriers for natural products such as plant oils and
extracts. ...",
publisher = "APGI – “Association de Pharmacie Galénique Industrielle”",
journal = "6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France",
title = "Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5189"
}
Gledović, A., Ilić, T.,& Savić, S.. (2023). Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test. in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France
APGI – “Association de Pharmacie Galénique Industrielle”..
https://hdl.handle.net/21.15107/rcub_farfar_5189
Gledović A, Ilić T, Savić S. Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test. in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5189 .
Gledović, Ana, Ilić, Tanja, Savić, Snežana, "Eye irritation potential of red raspberry seed oil after nanoemulsification: assessment with the HET-CAM test" in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5189 .

Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations

Ilić, Tanja; Gledović, Ana; Dobričić, Vladimir; Pantelić, Ivana; Savić, Snežana

(APGI – “Association de Pharmacie Galénique Industrielle”, 2023)

TY  - CONF
AU  - Ilić, Tanja
AU  - Gledović, Ana
AU  - Dobričić, Vladimir
AU  - Pantelić, Ivana
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5182
AB  - Repeated sun-exposure is one of the main sources of oxidative stress in the skin, which is responsible for the majority of age-associated skin conditions. ...
PB  - APGI – “Association de Pharmacie Galénique Industrielle”
C3  - 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France
T1  - Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5182
ER  - 
@conference{
author = "Ilić, Tanja and Gledović, Ana and Dobričić, Vladimir and Pantelić, Ivana and Savić, Snežana",
year = "2023",
abstract = "Repeated sun-exposure is one of the main sources of oxidative stress in the skin, which is responsible for the majority of age-associated skin conditions. ...",
publisher = "APGI – “Association de Pharmacie Galénique Industrielle”",
journal = "6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France",
title = "Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5182"
}
Ilić, T., Gledović, A., Dobričić, V., Pantelić, I.,& Savić, S.. (2023). Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations. in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France
APGI – “Association de Pharmacie Galénique Industrielle”..
https://hdl.handle.net/21.15107/rcub_farfar_5182
Ilić T, Gledović A, Dobričić V, Pantelić I, Savić S. Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations. in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_5182 .
Ilić, Tanja, Gledović, Ana, Dobričić, Vladimir, Pantelić, Ivana, Savić, Snežana, "Optimization of ORAC assay combined with in vivo tape stripping for the assessment of antioxidant efficacy of cosmetic formulations" in 6th Symposium Skin and Formulation, 2-3 October, 2023, Nantes - France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_5182 .

Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays

Nikolić, Ines; Petrović, Marija; Mitrović, Jelena; Sublet, Emmanuelle; Jordan, Oliver; Savić, Snežana; Borchard, Gerrit

(International Association of Physical Chemists, 2023)

TY  - CONF
AU  - Nikolić, Ines
AU  - Petrović, Marija
AU  - Mitrović, Jelena
AU  - Sublet, Emmanuelle
AU  - Jordan, Oliver
AU  - Savić, Snežana
AU  - Borchard, Gerrit
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/5007
AB  - It is well known that the characterization of nanomedicines can pursue different levels of complexity, both in the development stage and in the quality control process [1]. In line with physicochemical aspects, even more obstacles are encountered in biological safety assessment, while anticipation of their immunogenic potential represents an additional challenge. Moreover, interactions between the test reagents and the nanomaterial have been identified as one of the most important issues in toxicity testing that influence market authorization of nanomedicines, which ought to be resolved [1]. The European Nanomedicine Characterization Laboratory – the reference laboratory for nanomedicines, provides protocols for 2 colorimetric cytotoxicity assays employing LLC-PK1 (porcine kidney epithelial cells) and Hep-G2 (human hepatocarcinoma cells) cell lines. However, the latest recommendations in the field underline the demand for enhancing the testing procedures, while proposing incorporation of immune cells as target cell lines for toxicity evaluation, aiming to provide more reliable conclusions on nanomedicine safety in preclinical level.
In this study 2 inherently different types of pharmaceutical nanosystemes were selected: nanoemulsion (NE) and solid lipid nanoparticles (LNP) and subjected to a set of orthogonal toxicity evaluation assays. Adjusted WST-1 (assessing mitochondrial activity as an indicator of cellular well-being) and LDH (lactate dehydrogenase release evaluation as an indicator of cell membrane damage) assays have been performed as the colorimetric tests, while propidium-iodide (PI)-based assay was developed as a fluore-scent counterpart (able to directly distinguish live from dead cells), using RAW 246.7 cell line (murine macrophages – immune system cell line). Starting concentration of the tested nanoformulations was 50 % v/v, and they were subsequently diluted with the factor of 2, to create a total of 8 concentrations. Incubation time was 4 h.
Presented assays rely on completely different biological bases. Therefore, their careful combination can address some shortcomings in the in vitro evaluations established so far. Although similar toxicity trends were observed regardless the assay used, it was evident that the LDH assay required specific consideration. Since the supernatant is the subject of the analysis (not the cells directly), containing not only the enzyme of interest, but also the nanoformulations, in the wells corresponding to the 3 highest concentration of the NE/LNP pronounced scattering effects were observed. Such an event could be easily overlooked, potentially affecting the conclusions. However, it was overcome by careful design of control and blank wells (each test concentration was coupled with its own blank well containing no cells, but the same concentration of the NE/LNP in the culture medium). In contrast, absorbance measurements in WST-1 assay were performed in the absence of the NE/LNP, avoiding any interactions or scattering effects. Finally, developed PI-based assay proved to be the most relevant method. Based on the penetration of PI into the dead cell only, attaching to their DNA, the concentration of the dead vs. live cells could be directly estimated. What is more, after the incubation time, the measurements can be performed in the nanoformulation-free environment, surpas-sing the potential interactions. Notably, cell viability obtained in the PI-based assay followed the same trend as in the WST-1 assay but with significant difference in the obtained values for the first 3 concentrations (Figure 1).
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  - Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays
SP  - 21
EP  - 21
UR  - https://hdl.handle.net/21.15107/rcub_farfar_5007
ER  - 
@conference{
author = "Nikolić, Ines and Petrović, Marija and Mitrović, Jelena and Sublet, Emmanuelle and Jordan, Oliver and Savić, Snežana and Borchard, Gerrit",
year = "2023",
abstract = "It is well known that the characterization of nanomedicines can pursue different levels of complexity, both in the development stage and in the quality control process [1]. In line with physicochemical aspects, even more obstacles are encountered in biological safety assessment, while anticipation of their immunogenic potential represents an additional challenge. Moreover, interactions between the test reagents and the nanomaterial have been identified as one of the most important issues in toxicity testing that influence market authorization of nanomedicines, which ought to be resolved [1]. The European Nanomedicine Characterization Laboratory – the reference laboratory for nanomedicines, provides protocols for 2 colorimetric cytotoxicity assays employing LLC-PK1 (porcine kidney epithelial cells) and Hep-G2 (human hepatocarcinoma cells) cell lines. However, the latest recommendations in the field underline the demand for enhancing the testing procedures, while proposing incorporation of immune cells as target cell lines for toxicity evaluation, aiming to provide more reliable conclusions on nanomedicine safety in preclinical level.
In this study 2 inherently different types of pharmaceutical nanosystemes were selected: nanoemulsion (NE) and solid lipid nanoparticles (LNP) and subjected to a set of orthogonal toxicity evaluation assays. Adjusted WST-1 (assessing mitochondrial activity as an indicator of cellular well-being) and LDH (lactate dehydrogenase release evaluation as an indicator of cell membrane damage) assays have been performed as the colorimetric tests, while propidium-iodide (PI)-based assay was developed as a fluore-scent counterpart (able to directly distinguish live from dead cells), using RAW 246.7 cell line (murine macrophages – immune system cell line). Starting concentration of the tested nanoformulations was 50 % v/v, and they were subsequently diluted with the factor of 2, to create a total of 8 concentrations. Incubation time was 4 h.
Presented assays rely on completely different biological bases. Therefore, their careful combination can address some shortcomings in the in vitro evaluations established so far. Although similar toxicity trends were observed regardless the assay used, it was evident that the LDH assay required specific consideration. Since the supernatant is the subject of the analysis (not the cells directly), containing not only the enzyme of interest, but also the nanoformulations, in the wells corresponding to the 3 highest concentration of the NE/LNP pronounced scattering effects were observed. Such an event could be easily overlooked, potentially affecting the conclusions. However, it was overcome by careful design of control and blank wells (each test concentration was coupled with its own blank well containing no cells, but the same concentration of the NE/LNP in the culture medium). In contrast, absorbance measurements in WST-1 assay were performed in the absence of the NE/LNP, avoiding any interactions or scattering effects. Finally, developed PI-based assay proved to be the most relevant method. Based on the penetration of PI into the dead cell only, attaching to their DNA, the concentration of the dead vs. live cells could be directly estimated. What is more, after the incubation time, the measurements can be performed in the nanoformulation-free environment, surpas-sing the potential interactions. Notably, cell viability obtained in the PI-based assay followed the same trend as in the WST-1 assay but with significant difference in the obtained values for the first 3 concentrations (Figure 1).",
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 = "Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays",
pages = "21-21",
url = "https://hdl.handle.net/21.15107/rcub_farfar_5007"
}
Nikolić, I., Petrović, M., Mitrović, J., Sublet, E., Jordan, O., Savić, S.,& Borchard, G.. (2023). Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays. 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., 21-21.
https://hdl.handle.net/21.15107/rcub_farfar_5007
Nikolić I, Petrović M, Mitrović J, Sublet E, Jordan O, Savić S, Borchard G. Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays. 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;:21-21.
https://hdl.handle.net/21.15107/rcub_farfar_5007 .
Nikolić, Ines, Petrović, Marija, Mitrović, Jelena, Sublet, Emmanuelle, Jordan, Oliver, Savić, Snežana, Borchard, Gerrit, "Navigating towards improved cytotoxicity assessment in nanomedicine development: Shifting from colorimetric to fluorescence-based assays" 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):21-21,
https://hdl.handle.net/21.15107/rcub_farfar_5007 .

Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study

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

(Elsevier B.V., 2023)

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 . .

Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines

Filipić, Brankica; Pantelić, Ivana; Nikolić, Ines; Majhen, Dragomira; Stojić-Vukanić, Zorica; Savić, Snežana; Krajišnik, Danina

(MDPI, 2023)

TY  - JOUR
AU  - Filipić, Brankica
AU  - Pantelić, Ivana
AU  - Nikolić, Ines
AU  - Majhen, Dragomira
AU  - Stojić-Vukanić, Zorica
AU  - Savić, Snežana
AU  - Krajišnik, Danina
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4960
AB  - Ever since the development of the first vaccine, vaccination has had the great impact on global health, leading to the decrease in the burden of numerous infectious diseases. However, there is a constant need to improve existing vaccines and develop new vaccination strategies and vaccine platforms that induce a broader immune response compared to traditional vaccines. Modern vaccines tend to rely on certain nanotechnology platforms but are still expected to be readily available and easy for large-scale manufacturing and to induce a durable immune response. In this review, we present an overview of the most promising nanoadjuvants and nanoparticulate delivery systems and discuss their benefits from tehchnological and immunological standpoints as well as their objective drawbacks and possible side effects. The presented nano alums, silica and clay nanoparticles, nanoemulsions, adenoviral-vectored systems, adeno-associated viral vectors, vesicular stomatitis viral vectors, lentiviral vectors, virus-like particles (including bacteriophage-based ones) and virosomes indicate that vaccine developers can now choose different adjuvants and/or delivery systems as per the requirement, specific to combatting different infectious diseases.
PB  - MDPI
T2  - Vaccines
T1  - Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines
VL  - 11
IS  - 7
DO  - 10.3390/vaccines11071172
ER  - 
@article{
author = "Filipić, Brankica and Pantelić, Ivana and Nikolić, Ines and Majhen, Dragomira and Stojić-Vukanić, Zorica and Savić, Snežana and Krajišnik, Danina",
year = "2023",
abstract = "Ever since the development of the first vaccine, vaccination has had the great impact on global health, leading to the decrease in the burden of numerous infectious diseases. However, there is a constant need to improve existing vaccines and develop new vaccination strategies and vaccine platforms that induce a broader immune response compared to traditional vaccines. Modern vaccines tend to rely on certain nanotechnology platforms but are still expected to be readily available and easy for large-scale manufacturing and to induce a durable immune response. In this review, we present an overview of the most promising nanoadjuvants and nanoparticulate delivery systems and discuss their benefits from tehchnological and immunological standpoints as well as their objective drawbacks and possible side effects. The presented nano alums, silica and clay nanoparticles, nanoemulsions, adenoviral-vectored systems, adeno-associated viral vectors, vesicular stomatitis viral vectors, lentiviral vectors, virus-like particles (including bacteriophage-based ones) and virosomes indicate that vaccine developers can now choose different adjuvants and/or delivery systems as per the requirement, specific to combatting different infectious diseases.",
publisher = "MDPI",
journal = "Vaccines",
title = "Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines",
volume = "11",
number = "7",
doi = "10.3390/vaccines11071172"
}
Filipić, B., Pantelić, I., Nikolić, I., Majhen, D., Stojić-Vukanić, Z., Savić, S.,& Krajišnik, D.. (2023). Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines. in Vaccines
MDPI., 11(7).
https://doi.org/10.3390/vaccines11071172
Filipić B, Pantelić I, Nikolić I, Majhen D, Stojić-Vukanić Z, Savić S, Krajišnik D. Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines. in Vaccines. 2023;11(7).
doi:10.3390/vaccines11071172 .
Filipić, Brankica, Pantelić, Ivana, Nikolić, Ines, Majhen, Dragomira, Stojić-Vukanić, Zorica, Savić, Snežana, Krajišnik, Danina, "Nanoparticle-Based Adjuvants and Delivery Systems for Modern Vaccines" in Vaccines, 11, no. 7 (2023),
https://doi.org/10.3390/vaccines11071172 . .
5
5

Current role of tribological tests: striving for full characterization of medicinal and cosmetic products

Tošić, Anđela; Stanković, Tijana; Ilić, Tanja; Savić, Snežana; Pantelić, Ivana

(Savez farmaceutskih udruženja Srbije, Beograd, 2023)

TY  - JOUR
AU  - Tošić, Anđela
AU  - Stanković, Tijana
AU  - Ilić, Tanja
AU  - Savić, Snežana
AU  - Pantelić, Ivana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4770
AB  - Tribology investigates the events that happen on the surfaces of two substances/objects that 
are in direct or indirect contact through assessing friction, lubrication and/or wear. In particular, 
friction measurements could provide the information on the textural characteristics of (per)oral 
pharmaceutical  preparations  and  contribute  to  the  understanding  of  palatability.  On  the  other  
hand,  tribological  tests  have  been  more  intensively  used  to  characterize  topical  preparations  
(pharmaceutical,  cosmetic),  giving  a  thorough  insight  into  the  tactile  and  texture  properties  of  
these preparations. However, these tests are often combined with rheological, textural, and certain 
biophysical  approa
ches.  Additionally,  the  materials  used  for  constructing  artificial  joints  and  
articular  cartilages  are  true  tribological  systems,  developed  and  optimized  in  order  to  have  
properties that resemble the natural ones. Since tribological studies can be used to assess a wide 
range of drug dosage forms and products in general, the equipment used may be quite diverse. 
Accordingly,  a  special  section  of  this  work  is  committed  to  the  description  of  the  testing  
equipment’s  specifications  and  the  applied  protocols.  The  investigation  of  recently  regulatory  
discovered  phenomena,  such  as  transformation/metamorphosis  of  the  vehicle/base  of  topical  
preparations, have brought tribology back into focus as a potential assessment method.
AB  - Tribologija se bavi izučavanjem uticaja i 
događaja koji se dešavaju na površinama dveju 
materija/objekata  koji  su  u  direktnom  ili  indirektnom  kontaktu,  uključujući  procese  trenja, 
podmazivanja i/ili habanja (trošenja). Naime, primećeno je da je merenjem frikcije moguće 
ispitati teksturna svojstva 
(per)oralnih farmaceutskih preparata i doprineti razumevanju njihove 
palatabilnosti.  S  druge  strane,  nešto  duži  niz  godina  se  tribološka  ispitivanja  izvode  u  cilju  
karakterizacije preparata (farmaceutskih, kozmetičkih) koji se primenjuju na koži, dajući ti
me 
kompletniju sliku o taktilnim i teksturnim osobinama ovih preparata. Ipak, dobijeni rezultati se 
uobičajeno razmatraju zajedno sa onim dobijenim tokom reoloških, teksturnih i/ili biofizičkih 
studija. Takođe, materijali od kojih su napravljeni veštački z
globovi i zglobne hrskavice su primer 
triboloških  sistema  koji  su  razvijani  i  optimizovani  na  način  da  imaju  slične  karakteristike 
prirodnim sistemima, a za čiji je razvoj i karakterizaciju neophodno ispitivanje frikcije, lubrikacije 
i trošenja. Kako je po
lje primene triboloških ispitivanja široko i provlači se kroz, u tehnološkom 
smislu, izrazito različite farmaceutske oblike, posledično će i aparatura koja se koristi u te svrhe 
pokazivati veliki diverzitet, te je deo ovog rada posvećen i pregledu specifič
nosti uređaja za 
ispitivanje triboloških parametara, sa posebnim osvrtom na primenjene protokole ispitivanja. Na 
kraju,  dat  je  osvrt  na  potencijalne  primene  triboloških  studija  za  ispitivanje  novootkrivenih  
fenomena, poput transformacije/metamorfoze vehikuluma/podloge topikalnih preparata.
PB  - Savez farmaceutskih udruženja Srbije, Beograd
T2  - Arhiv za farmaciju
T1  - Current role of tribological tests: striving for full  characterization of medicinal and cosmetic  products
T1  - Doprinos triboloških testova sveobuhvatnoj 
karakterizaciji medicinskih i kozmetičkih 
proizvoda
VL  - 73
IS  - 2
SP  - 126
EP  - 145
DO  - 10.5937/arhfarm73- 43515
ER  - 
@article{
author = "Tošić, Anđela and Stanković, Tijana and Ilić, Tanja and Savić, Snežana and Pantelić, Ivana",
year = "2023",
abstract = "Tribology investigates the events that happen on the surfaces of two substances/objects that 
are in direct or indirect contact through assessing friction, lubrication and/or wear. In particular, 
friction measurements could provide the information on the textural characteristics of (per)oral 
pharmaceutical  preparations  and  contribute  to  the  understanding  of  palatability.  On  the  other  
hand,  tribological  tests  have  been  more  intensively  used  to  characterize  topical  preparations  
(pharmaceutical,  cosmetic),  giving  a  thorough  insight  into  the  tactile  and  texture  properties  of  
these preparations. However, these tests are often combined with rheological, textural, and certain 
biophysical  approa
ches.  Additionally,  the  materials  used  for  constructing  artificial  joints  and  
articular  cartilages  are  true  tribological  systems,  developed  and  optimized  in  order  to  have  
properties that resemble the natural ones. Since tribological studies can be used to assess a wide 
range of drug dosage forms and products in general, the equipment used may be quite diverse. 
Accordingly,  a  special  section  of  this  work  is  committed  to  the  description  of  the  testing  
equipment’s  specifications  and  the  applied  protocols.  The  investigation  of  recently  regulatory  
discovered  phenomena,  such  as  transformation/metamorphosis  of  the  vehicle/base  of  topical  
preparations, have brought tribology back into focus as a potential assessment method., Tribologija se bavi izučavanjem uticaja i 
događaja koji se dešavaju na površinama dveju 
materija/objekata  koji  su  u  direktnom  ili  indirektnom  kontaktu,  uključujući  procese  trenja, 
podmazivanja i/ili habanja (trošenja). Naime, primećeno je da je merenjem frikcije moguće 
ispitati teksturna svojstva 
(per)oralnih farmaceutskih preparata i doprineti razumevanju njihove 
palatabilnosti.  S  druge  strane,  nešto  duži  niz  godina  se  tribološka  ispitivanja  izvode  u  cilju  
karakterizacije preparata (farmaceutskih, kozmetičkih) koji se primenjuju na koži, dajući ti
me 
kompletniju sliku o taktilnim i teksturnim osobinama ovih preparata. Ipak, dobijeni rezultati se 
uobičajeno razmatraju zajedno sa onim dobijenim tokom reoloških, teksturnih i/ili biofizičkih 
studija. Takođe, materijali od kojih su napravljeni veštački z
globovi i zglobne hrskavice su primer 
triboloških  sistema  koji  su  razvijani  i  optimizovani  na  način  da  imaju  slične  karakteristike 
prirodnim sistemima, a za čiji je razvoj i karakterizaciju neophodno ispitivanje frikcije, lubrikacije 
i trošenja. Kako je po
lje primene triboloških ispitivanja široko i provlači se kroz, u tehnološkom 
smislu, izrazito različite farmaceutske oblike, posledično će i aparatura koja se koristi u te svrhe 
pokazivati veliki diverzitet, te je deo ovog rada posvećen i pregledu specifič
nosti uređaja za 
ispitivanje triboloških parametara, sa posebnim osvrtom na primenjene protokole ispitivanja. Na 
kraju,  dat  je  osvrt  na  potencijalne  primene  triboloških  studija  za  ispitivanje  novootkrivenih  
fenomena, poput transformacije/metamorfoze vehikuluma/podloge topikalnih preparata.",
publisher = "Savez farmaceutskih udruženja Srbije, Beograd",
journal = "Arhiv za farmaciju",
title = "Current role of tribological tests: striving for full  characterization of medicinal and cosmetic  products, Doprinos triboloških testova sveobuhvatnoj 
karakterizaciji medicinskih i kozmetičkih 
proizvoda",
volume = "73",
number = "2",
pages = "126-145",
doi = "10.5937/arhfarm73- 43515"
}
Tošić, A., Stanković, T., Ilić, T., Savić, S.,& Pantelić, I.. (2023). Current role of tribological tests: striving for full  characterization of medicinal and cosmetic  products. in Arhiv za farmaciju
Savez farmaceutskih udruženja Srbije, Beograd., 73(2), 126-145.
https://doi.org/10.5937/arhfarm73- 43515
Tošić A, Stanković T, Ilić T, Savić S, Pantelić I. Current role of tribological tests: striving for full  characterization of medicinal and cosmetic  products. in Arhiv za farmaciju. 2023;73(2):126-145.
doi:10.5937/arhfarm73- 43515 .
Tošić, Anđela, Stanković, Tijana, Ilić, Tanja, Savić, Snežana, Pantelić, Ivana, "Current role of tribological tests: striving for full  characterization of medicinal and cosmetic  products" in Arhiv za farmaciju, 73, no. 2 (2023):126-145,
https://doi.org/10.5937/arhfarm73- 43515 . .

Contribution of various instrumental methods to transformation/metamorphosis assessment of hydrophilic gels during skin application

Tošić, Anđela; Ilić, Tanja; Savić, Snežana; Pantelić, Ivana

(2023)

TY  - CONF
AU  - Tošić, Anđela
AU  - Ilić, Tanja
AU  - Savić, Snežana
AU  - Pantelić, Ivana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4592
AB  - INTRODUCTION The metamorphosis of topical products is a relatively new concept that has grown in importance during the past few years [1]. It is defined by the phenomenon in which the primary packaged formulation changes its composition and/or microstructure during application. This change could be caused by different internal/external stimuli, most commonly, but not limited to the evaporation of highly volatile ingredients. As a consequence, there is a high possibility of a discrepancy between the desired release profile of the active substance and the obtained one. In addition to the pharmacokinetic profile changes, metamorphosis can also cause changes in the rheological, texture and cosmetic properties of the preparation [2,3]. According to the most recent EMA Draft Guideline on quality and equivalence of topical products from 2018, it is necessary to examine metamorphosis when assessing the bioequivalence of (trans)dermal products [1]. At this moment, a part of the scientific community suggests only two methods for metamorphosis assessment: nanothermal analysis and photothermal microspectroscopy. Both methods use probes to detect crystals of the drug substance in the deeper layers of the stratum corneum [4]. However, these methods require very expensive, sophisticated equipment, and they have not yet been formalized as methodologies used for this purpose. The aim of this work was to evaluate the possibility of using rheological, tribological and mass loss testing as single or combined methods for the assessment of metamorphosis of topical hydrogels. MATERIALS AND METHODS Materials Carbopol® 934 was obtained from Lubrizol (USA), while propylene glycol, triethanolamine, isopropanol, sodium hydroxide, methyl and propyl paraben were purchased from Sigma Aldrich (Germany). A model substance, diclofenac-sodium, was kindly donated by Hemofarm (Serbia). Sample preparation The concentration of isopropanol, as a model easily volatile ingredient, was varied in the range 0-15% (w/w). In the sample F1 (Table 1), the concentration of the gelling agent was also varied, as another parameter of interest for valid transformation analysis. The gels were prepared in accordance with the usual compendial guidelines (DAC/NRF). Rheological characterization Measurements were preformed using a Rheolab MC 120 rotational rheometer (Paar Physica, Germany), with a cone/plate system with a diameter of 50 mm, at an angle of 1° and a sample thickness of 0.05 mm, at a temperature of 20±0.1°C. Tribological study Test was performed in vivo in 4 female healthy volunteers, at the forearm skin, using Frictiometer® FR700 (Courage+Khazaka, Germany) equipped with a plain, smooth Teflon (PTFE) disk. Study was performed both in finite and infinite dosing conditions. Measurements were performed at 90 rpm for 100 s, with one measurement taken per second. Mass loss analysis The test was performed at a temperature of 32±0.1°C in a closed system of the device Orbital Shaker Incubator ES 20 (Biosan, Latvia). Each sample was applied in the quantity of 1 g, in a thin layer on the glass substrate and placed in a closed chamber system. The mass of the samples was measured on the ABJ 120-4M analytical scale (Kern & Sohn GmbH, Germany) during two hours in 15-minute intervals. RESULTS AND DISCUSSION The greatest potential for instrumental transformation analysis has been demonstrated by the simple mass loss study. In Figure 1, it can be clearly seen that samples F3-F6, which contain isopropanol, have reached the "plateau" sooner than samples F1-F2. The slope of the curve (transformation rate) between 45 and 60 min of drying is also significantly different for the samples F1- F2 and F3-F6. This method additionally allows discerning regions that represent the secondary and third (residual) formulations. Part of the curve between 15 and 45 min, when the mass loss is the most significant, correlates with forming the secondary formulation. The region between 45 and 120 min, depending on the very sample, represents the process of forming the residual formulation, after all the volatile ingredients have evaporated. On the other hand, rheological test has shown somewhat different results. Flow curves (Figure 2) represent changes in the microstructure that formulations go through during the test. Expectedly, gels that contain isopropanol go through more drastic changes. In the descending part of the curve unsymmetrical regions could be spotted, relative to the ascending part. This phenomenon could not be noted in the flow curves of the formulations F1 and F2. Furthermore, maximal and minimal viscosities and hysteresis area were the parameters that failed to show great potential for in depth assessment of a vehicle’s metamorphosis.A tribological study carried out under finite dosing conditions (3 mg/cm2 ; Figure 3a) provided more informative results, especially for the samples’ F3-F6 process of metamorphosis. The levels of changes in the friction value correlate well with the concentration of isopropyl alcohol. Therefore, it can be seen that the changes of this parameter are more intensive for formulations F5-F6 compared to formulations F3-F4, that contained isopropyl alcohol in lower concentrations. The same study, performed under infinite dosing conditions (10 mg/cm2 ; Figure 3b), showed no apparent potential for these purposes.CONSLUSION The results have showed that all three methods in a certain sense contribute to the examination of the metamorphosis of carbomer gels. Certain time points during mass loss and friction tests correlate well in terms of the exact onset of each transformation phase.
C3  - 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France
T1  - Contribution of various instrumental methods to  transformation/metamorphosis assessment of  hydrophilic gels during skin application
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4592
ER  - 
@conference{
author = "Tošić, Anđela and Ilić, Tanja and Savić, Snežana and Pantelić, Ivana",
year = "2023",
abstract = "INTRODUCTION The metamorphosis of topical products is a relatively new concept that has grown in importance during the past few years [1]. It is defined by the phenomenon in which the primary packaged formulation changes its composition and/or microstructure during application. This change could be caused by different internal/external stimuli, most commonly, but not limited to the evaporation of highly volatile ingredients. As a consequence, there is a high possibility of a discrepancy between the desired release profile of the active substance and the obtained one. In addition to the pharmacokinetic profile changes, metamorphosis can also cause changes in the rheological, texture and cosmetic properties of the preparation [2,3]. According to the most recent EMA Draft Guideline on quality and equivalence of topical products from 2018, it is necessary to examine metamorphosis when assessing the bioequivalence of (trans)dermal products [1]. At this moment, a part of the scientific community suggests only two methods for metamorphosis assessment: nanothermal analysis and photothermal microspectroscopy. Both methods use probes to detect crystals of the drug substance in the deeper layers of the stratum corneum [4]. However, these methods require very expensive, sophisticated equipment, and they have not yet been formalized as methodologies used for this purpose. The aim of this work was to evaluate the possibility of using rheological, tribological and mass loss testing as single or combined methods for the assessment of metamorphosis of topical hydrogels. MATERIALS AND METHODS Materials Carbopol® 934 was obtained from Lubrizol (USA), while propylene glycol, triethanolamine, isopropanol, sodium hydroxide, methyl and propyl paraben were purchased from Sigma Aldrich (Germany). A model substance, diclofenac-sodium, was kindly donated by Hemofarm (Serbia). Sample preparation The concentration of isopropanol, as a model easily volatile ingredient, was varied in the range 0-15% (w/w). In the sample F1 (Table 1), the concentration of the gelling agent was also varied, as another parameter of interest for valid transformation analysis. The gels were prepared in accordance with the usual compendial guidelines (DAC/NRF). Rheological characterization Measurements were preformed using a Rheolab MC 120 rotational rheometer (Paar Physica, Germany), with a cone/plate system with a diameter of 50 mm, at an angle of 1° and a sample thickness of 0.05 mm, at a temperature of 20±0.1°C. Tribological study Test was performed in vivo in 4 female healthy volunteers, at the forearm skin, using Frictiometer® FR700 (Courage+Khazaka, Germany) equipped with a plain, smooth Teflon (PTFE) disk. Study was performed both in finite and infinite dosing conditions. Measurements were performed at 90 rpm for 100 s, with one measurement taken per second. Mass loss analysis The test was performed at a temperature of 32±0.1°C in a closed system of the device Orbital Shaker Incubator ES 20 (Biosan, Latvia). Each sample was applied in the quantity of 1 g, in a thin layer on the glass substrate and placed in a closed chamber system. The mass of the samples was measured on the ABJ 120-4M analytical scale (Kern & Sohn GmbH, Germany) during two hours in 15-minute intervals. RESULTS AND DISCUSSION The greatest potential for instrumental transformation analysis has been demonstrated by the simple mass loss study. In Figure 1, it can be clearly seen that samples F3-F6, which contain isopropanol, have reached the "plateau" sooner than samples F1-F2. The slope of the curve (transformation rate) between 45 and 60 min of drying is also significantly different for the samples F1- F2 and F3-F6. This method additionally allows discerning regions that represent the secondary and third (residual) formulations. Part of the curve between 15 and 45 min, when the mass loss is the most significant, correlates with forming the secondary formulation. The region between 45 and 120 min, depending on the very sample, represents the process of forming the residual formulation, after all the volatile ingredients have evaporated. On the other hand, rheological test has shown somewhat different results. Flow curves (Figure 2) represent changes in the microstructure that formulations go through during the test. Expectedly, gels that contain isopropanol go through more drastic changes. In the descending part of the curve unsymmetrical regions could be spotted, relative to the ascending part. This phenomenon could not be noted in the flow curves of the formulations F1 and F2. Furthermore, maximal and minimal viscosities and hysteresis area were the parameters that failed to show great potential for in depth assessment of a vehicle’s metamorphosis.A tribological study carried out under finite dosing conditions (3 mg/cm2 ; Figure 3a) provided more informative results, especially for the samples’ F3-F6 process of metamorphosis. The levels of changes in the friction value correlate well with the concentration of isopropyl alcohol. Therefore, it can be seen that the changes of this parameter are more intensive for formulations F5-F6 compared to formulations F3-F4, that contained isopropyl alcohol in lower concentrations. The same study, performed under infinite dosing conditions (10 mg/cm2 ; Figure 3b), showed no apparent potential for these purposes.CONSLUSION The results have showed that all three methods in a certain sense contribute to the examination of the metamorphosis of carbomer gels. Certain time points during mass loss and friction tests correlate well in terms of the exact onset of each transformation phase.",
journal = "4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France",
title = "Contribution of various instrumental methods to  transformation/metamorphosis assessment of  hydrophilic gels during skin application",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4592"
}
Tošić, A., Ilić, T., Savić, S.,& Pantelić, I.. (2023). Contribution of various instrumental methods to  transformation/metamorphosis assessment of  hydrophilic gels during skin application. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France.
https://hdl.handle.net/21.15107/rcub_farfar_4592
Tošić A, Ilić T, Savić S, Pantelić I. Contribution of various instrumental methods to  transformation/metamorphosis assessment of  hydrophilic gels during skin application. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_4592 .
Tošić, Anđela, Ilić, Tanja, Savić, Snežana, Pantelić, Ivana, "Contribution of various instrumental methods to  transformation/metamorphosis assessment of  hydrophilic gels during skin application" in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_4592 .

Sizing experiments and bio-nano interactions: method matters

Nikolić, Ines; Petrović, Marija; Krupnik, Leondard; Ranđelović, Danijela; Avaro, Jonathan; Neels, Antonia; Borchard, Gerrit; Jordan, Olivier; Đoković, Jelena; Savić, Snežana

(2023)

TY  - CONF
AU  - Nikolić, Ines
AU  - Petrović, Marija
AU  - Krupnik, Leondard
AU  - Ranđelović, Danijela
AU  - Avaro, Jonathan
AU  - Neels, Antonia
AU  - Borchard, Gerrit
AU  - Jordan, Olivier
AU  - Đoković, Jelena
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4571
AB  - INTRODUCTION
Physicochemical properties of many active ingredients jeopardize their pharmacological activity. To overcome identified obstacles, nanosystems as carriers for delivery of actives have been recognized as promising tools. Increasing number of applications for registration of nanotechnology-enabled pharmaceuticals and many more currently in preclinical or clinical studies raised some questions not only in the field of research and development, but also for regulators. Given the complexity of nanosystems, some specific challenges have been encountered in their characterization, which have not been fully addressed despite respectable research tradition in this field.  Particle size and aggregation potential (especially in complex biological fluids) are some of the critical quality attributes of nanomedicines, being important in the context of physical stability of the colloidal system, and in terms of its safety profile and in vivo performance. Even though a bright future has been predicted for nanomedicines, some of the posted expectations have not been fully met so far. This might be reflected, at least at some points, in the certain methodological issues that commonly result in in vitro to in vivo translational gaps. This aspect underlines the importance of quality and safety assessment of nanomedicines which has also been recognized by globally leading research and regulatory bodies [1,2]. Therefore, the aim of the presented research was to perform a thorough analysis of the selected nanosystem (nanoemulsion) focusing on size estimation and particle-protein interaction applying several techniques, highlighting important factors for a reliable analysis.
METHODLOGY
Materials
As a model nanosystem, previously developed nanoemulsion was used, containing medium-chain triglycerides (Mygliol 812, Fagron) as the oil phase, combination of polysorbate 80 (Acros Organics) and soybean lecithin (Lipoid S-75, Lipoid) as stabilizes, and highly purified water as the water phase. For protein interaction assessment, human serum albumin was used (HSA, Sigma Aldrich).
Methods
Nanoemulsion preparation
Nanoemulsion was prepared via spontaneous emulsification, by dropwise addition of the mixture of the oil and stabilizers to the water phase under constant stirring. For nanoparticle-protein interaction assesment, nanoemulsion was incubated (1h, 37 °C) with HSA in the final concentration of 2.5 mg/ml. 
Sizing experiments – dynamic light scattering
Size and size distribution (per se and in biorelevant environment) were evaluated applying batch mode dynamic light scattering (DLS, Zetasizer Nano ZS90, Malvern Instruments, UK), following the NCL guidance [3]. Intensity-based average hydrodynamic diameter (Z-ave) and polydispersity index (PDI) were analysed in line with relevant parameters of the method. 
Atomic force microscopy (AFM)
Additional sizing analysis and morphological evaluation of the sample were performed applying AFM as a high-resolution technique. AFM analysis of the samples was performed by NTEGRA Prima atomic force microscope (NT-MDT, Moscow, Russia). Intermittent-contact AFM mode was applied using NT-MDT NSGO1 silicon, N-type, antimony doped cantilevers with Au reflective coating. Sample dilution corresponded to the optimal one selected for DLS, and 10 μl of the dilution was placed to the high-quality silica discs (Highest Grade V1 Mica Discs, Ted Pella Inc.) and dried in vacuum. Experiments were performed in the air, in contactless mode. Topographic images and “signal-error” images were collected, AFM images were created and analyzed with the software Image Analysis 2.2.0 (NT-MDT) and Gwyddion 2.60 (Free and Open Source software, Department of Nanometrology, Czech Metrology Institute).
Small angle X-ray scattering (SAXS)
SAXS experiments were performed with the general idea to analyze the structure of the dispersed nanodroplets more profoundly, and especially interactions in biorelevant surrounding (in contact with HSA). A laboratory X-ray setup was applied (Bruker Nanostar, Bruker AXS GmbH, Karlsruhe, Germany). Here, the Kα-line of a X-ray Cu source with a wavelength of 1.541 Å was used and further monochromated by a X-ray mirror. The beam was collimated to a beam diameter of approximately 0.4 mm using three pinholes. The sample-detector distance was set to 107 cm, which lead to a q-range of 0.07 ≤ q ≤ 2.3 nm-1. Calibration of the scattering vector q and estimation of the instrumental resolution of Δq = 0.25 nm-1 was done by measuring the first diffraction peak of a silver behenate sample. The scattered intensity was measured with an avalanche-based detector (VÅNTEC-2000, Bruker AXS). The transmitted part of the beam was determined using a home-made semi-transparent beam stop. The scattered intensity was extracted, radially averaged and integrated over all q-values using the Bruker software DIF-FRAC.EVA (Bruker AXS, version 4.1). The 1D data was transmission corrected and then background subtracted from the scattering of the solvent and the capillary using Matlab 2022.
RESULTS AND DISCUSSION
When applying DLS, as a preliminary technique, primary attention was put on the selection of optimal dilution level for the measurement, analyzing attenuation factor, count rate and intercept of the correlation function in different dilution ratios and with different dilution media (water, PBS 7.4 and 10 mM NaCl), and dilution 1:100 (v/v) was marked as the optimal one. However, significant differences in obtained nanodroplet size was observed depending on the type of medium. When water was used as a dilution medium, significantly higher Z-ave values were obtained (83.71±0.86 nm) compared to the situations where PBS 7.4 (73.50±0.75nm) or 10 mM NaCl (76.59±0.50nm) were used as dilution medium, indicating how sample preparation protocol might be crucial. Even though DLS was not sensitive enough to detect any interaction with HSA (no significant difference in terms of Z-ave and PDI compared to the results obtained in the same dilution medium without HSA), AFM captured qualitative difference in the droplet topography (Figure 1), raising ides on nanoemulsion interfacial interaction with HSA and increased aggregation potential. Further on, SAXS confirmed the existence of a bilayer structure as indicated by a prominent correlation peak at around 1 nm-1, which corresponds to a bilayer thickness of around 6.2 nm. SAXS (Figure 2; probably corresponding to the lecithin formations at the interface). It may be assumed that the bilayer structure changes its structure when mixed with HSA. 
CONCLUSION
In this research, it has been demonstrated how important is to carefully select measurement conditions even for DLS -commonly used and the only standardized methods, in order to keep the measurements meaningful. Further on, not every method is capable of detecting some specific bio-nano interactions. Aiming to generate reliable datasets, condition sine qua non is to perform complementary techniques with increasing complexity. Further experimental segments should cover additional evaluation (e.g. analytical ultracentrifugation, thermal analysis, interfacial properties assessment, electron microscopy) that would shed light on bio-nano interactions important for in vivo fate of the nanosystems.
C3  - 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France
T1  - Sizing experiments and bio-nano interactions: method matters
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4571
ER  - 
@conference{
author = "Nikolić, Ines and Petrović, Marija and Krupnik, Leondard and Ranđelović, Danijela and Avaro, Jonathan and Neels, Antonia and Borchard, Gerrit and Jordan, Olivier and Đoković, Jelena and Savić, Snežana",
year = "2023",
abstract = "INTRODUCTION
Physicochemical properties of many active ingredients jeopardize their pharmacological activity. To overcome identified obstacles, nanosystems as carriers for delivery of actives have been recognized as promising tools. Increasing number of applications for registration of nanotechnology-enabled pharmaceuticals and many more currently in preclinical or clinical studies raised some questions not only in the field of research and development, but also for regulators. Given the complexity of nanosystems, some specific challenges have been encountered in their characterization, which have not been fully addressed despite respectable research tradition in this field.  Particle size and aggregation potential (especially in complex biological fluids) are some of the critical quality attributes of nanomedicines, being important in the context of physical stability of the colloidal system, and in terms of its safety profile and in vivo performance. Even though a bright future has been predicted for nanomedicines, some of the posted expectations have not been fully met so far. This might be reflected, at least at some points, in the certain methodological issues that commonly result in in vitro to in vivo translational gaps. This aspect underlines the importance of quality and safety assessment of nanomedicines which has also been recognized by globally leading research and regulatory bodies [1,2]. Therefore, the aim of the presented research was to perform a thorough analysis of the selected nanosystem (nanoemulsion) focusing on size estimation and particle-protein interaction applying several techniques, highlighting important factors for a reliable analysis.
METHODLOGY
Materials
As a model nanosystem, previously developed nanoemulsion was used, containing medium-chain triglycerides (Mygliol 812, Fagron) as the oil phase, combination of polysorbate 80 (Acros Organics) and soybean lecithin (Lipoid S-75, Lipoid) as stabilizes, and highly purified water as the water phase. For protein interaction assessment, human serum albumin was used (HSA, Sigma Aldrich).
Methods
Nanoemulsion preparation
Nanoemulsion was prepared via spontaneous emulsification, by dropwise addition of the mixture of the oil and stabilizers to the water phase under constant stirring. For nanoparticle-protein interaction assesment, nanoemulsion was incubated (1h, 37 °C) with HSA in the final concentration of 2.5 mg/ml. 
Sizing experiments – dynamic light scattering
Size and size distribution (per se and in biorelevant environment) were evaluated applying batch mode dynamic light scattering (DLS, Zetasizer Nano ZS90, Malvern Instruments, UK), following the NCL guidance [3]. Intensity-based average hydrodynamic diameter (Z-ave) and polydispersity index (PDI) were analysed in line with relevant parameters of the method. 
Atomic force microscopy (AFM)
Additional sizing analysis and morphological evaluation of the sample were performed applying AFM as a high-resolution technique. AFM analysis of the samples was performed by NTEGRA Prima atomic force microscope (NT-MDT, Moscow, Russia). Intermittent-contact AFM mode was applied using NT-MDT NSGO1 silicon, N-type, antimony doped cantilevers with Au reflective coating. Sample dilution corresponded to the optimal one selected for DLS, and 10 μl of the dilution was placed to the high-quality silica discs (Highest Grade V1 Mica Discs, Ted Pella Inc.) and dried in vacuum. Experiments were performed in the air, in contactless mode. Topographic images and “signal-error” images were collected, AFM images were created and analyzed with the software Image Analysis 2.2.0 (NT-MDT) and Gwyddion 2.60 (Free and Open Source software, Department of Nanometrology, Czech Metrology Institute).
Small angle X-ray scattering (SAXS)
SAXS experiments were performed with the general idea to analyze the structure of the dispersed nanodroplets more profoundly, and especially interactions in biorelevant surrounding (in contact with HSA). A laboratory X-ray setup was applied (Bruker Nanostar, Bruker AXS GmbH, Karlsruhe, Germany). Here, the Kα-line of a X-ray Cu source with a wavelength of 1.541 Å was used and further monochromated by a X-ray mirror. The beam was collimated to a beam diameter of approximately 0.4 mm using three pinholes. The sample-detector distance was set to 107 cm, which lead to a q-range of 0.07 ≤ q ≤ 2.3 nm-1. Calibration of the scattering vector q and estimation of the instrumental resolution of Δq = 0.25 nm-1 was done by measuring the first diffraction peak of a silver behenate sample. The scattered intensity was measured with an avalanche-based detector (VÅNTEC-2000, Bruker AXS). The transmitted part of the beam was determined using a home-made semi-transparent beam stop. The scattered intensity was extracted, radially averaged and integrated over all q-values using the Bruker software DIF-FRAC.EVA (Bruker AXS, version 4.1). The 1D data was transmission corrected and then background subtracted from the scattering of the solvent and the capillary using Matlab 2022.
RESULTS AND DISCUSSION
When applying DLS, as a preliminary technique, primary attention was put on the selection of optimal dilution level for the measurement, analyzing attenuation factor, count rate and intercept of the correlation function in different dilution ratios and with different dilution media (water, PBS 7.4 and 10 mM NaCl), and dilution 1:100 (v/v) was marked as the optimal one. However, significant differences in obtained nanodroplet size was observed depending on the type of medium. When water was used as a dilution medium, significantly higher Z-ave values were obtained (83.71±0.86 nm) compared to the situations where PBS 7.4 (73.50±0.75nm) or 10 mM NaCl (76.59±0.50nm) were used as dilution medium, indicating how sample preparation protocol might be crucial. Even though DLS was not sensitive enough to detect any interaction with HSA (no significant difference in terms of Z-ave and PDI compared to the results obtained in the same dilution medium without HSA), AFM captured qualitative difference in the droplet topography (Figure 1), raising ides on nanoemulsion interfacial interaction with HSA and increased aggregation potential. Further on, SAXS confirmed the existence of a bilayer structure as indicated by a prominent correlation peak at around 1 nm-1, which corresponds to a bilayer thickness of around 6.2 nm. SAXS (Figure 2; probably corresponding to the lecithin formations at the interface). It may be assumed that the bilayer structure changes its structure when mixed with HSA. 
CONCLUSION
In this research, it has been demonstrated how important is to carefully select measurement conditions even for DLS -commonly used and the only standardized methods, in order to keep the measurements meaningful. Further on, not every method is capable of detecting some specific bio-nano interactions. Aiming to generate reliable datasets, condition sine qua non is to perform complementary techniques with increasing complexity. Further experimental segments should cover additional evaluation (e.g. analytical ultracentrifugation, thermal analysis, interfacial properties assessment, electron microscopy) that would shed light on bio-nano interactions important for in vivo fate of the nanosystems.",
journal = "4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France",
title = "Sizing experiments and bio-nano interactions: method matters",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4571"
}
Nikolić, I., Petrović, M., Krupnik, L., Ranđelović, D., Avaro, J., Neels, A., Borchard, G., Jordan, O., Đoković, J.,& Savić, S.. (2023). Sizing experiments and bio-nano interactions: method matters. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France.
https://hdl.handle.net/21.15107/rcub_farfar_4571
Nikolić I, Petrović M, Krupnik L, Ranđelović D, Avaro J, Neels A, Borchard G, Jordan O, Đoković J, Savić S. Sizing experiments and bio-nano interactions: method matters. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_4571 .
Nikolić, Ines, Petrović, Marija, Krupnik, Leondard, Ranđelović, Danijela, Avaro, Jonathan, Neels, Antonia, Borchard, Gerrit, Jordan, Olivier, Đoković, Jelena, Savić, Snežana, "Sizing experiments and bio-nano interactions: method matters" in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_4571 .

Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer

Đoković, Jelena; Demisli, Sotiria; Papadimitriou, Vassiliki; Savić, Snežana

(2023)

TY  - CONF
AU  - Đoković, Jelena
AU  - Demisli, Sotiria
AU  - Papadimitriou, Vassiliki
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4572
AB  - INTRODUCTION
Nanoemulsions (NEs) offer a flexible platform for drug delivery via several administration routes. Rapid plasma clearance brought on by interactions with plasma proteins and the activation of the mononuclear phagocytic system is the greatest challenge NEs face after parenteral administration. PEGylation, or adding PEGylated phospholipids to the stabilizing layer of the NEs, is one method for ensuring that droplets circulate for a longer period of time. It is crucial to select the optimum concentration of the PEGylated in order to maintain the necessary physicochemical properties of NEs while providing appropriate surface coverage with the PEG chains. Curcumin is a model active that has been found to be localized in the stabilizing layer of NEs (1-3) and offers a wide range of potential health benefits, but due to its short plasma half-life, new strategies for enhancing bioavailability are required. The purpose of this study is to investigate the effects of various PEGylated phospholipid (PEG2000-DSPE) concentrations on the structural properties of NEs with an active placed in the stabilizing layer.
PREPARATION OF NANOEMULSIONS
All the NEs were prepared using the high pressure homogenization technique. The aqueous phase (glycerol, polysorbate 80, sodium oleate and highly purified water) was added to the oil phase (soybean oil, medium chain triglycerides, soybean lecithin, buthylhydroxytoluene, curcumin and 0.1%/0.3%/0.6% PEG2000-DSPE) and mixed at 11000rpm for 1 min on rotor stator homogenizer (IKA Ultra-Turrax T25 digital, IKA-Werke GmbH & Co. KG, Staufen, Germany), and then further processed at 800 bar for 10 discontinued cycles (EmulsiFlex-C3, Avestin Inc., Ottawa,ON, Canada) to obtain CS21, CS23 and CS26 formulations.
NANOEMULSION DROPLET SIZE
The droplet size was assessed through the dynamic light scattering method and presented as mean droplet size (Z-ave) and polydispersity index (PDI), after diluting the NEs 1:500 (v/v) in highly purified water.
ELECTRON PARAMAGNETIC RESONANCE (EPR) SPECTROSCOPY
For this study three amphiphilic fatty acid derivatives labeled at different positions of the aliphatic chain (5-DSA, 12-DSA and 16-DSA) were used to probe the dynamics of the membrane at different depths. Stock solutions of the spin probes were prepared in absolute ethanol at 1mM concentration. Subsequently, 15 µl of the stock solutions were evaporated and then incubated with 1 ml of the NE sample in final concentration of 0.015 mM. The resulting spectra was analyzed in terms of rotational correlation time (τR), order parameter (S) and isotropic hyperfine constant (αN).
RESULTS AND DISCUSION
All of the formulations had average droplet sizes between 95 and 103 nm and PDI values under 0.25, which indicated that they were suitable for parenteral administration.
The results of the EPR investigation showed that the stabilizing layer changed as the amount of PEGylated phospholipids increased, indicating that the PEGylation threshold has not yet been reached in the stabilizing layer.
The EPR research also showed that the 5-DSA spin probe's spectra were significantly affected by the addition of various PEGylated phospholipid concentrations (Figure 1). This indicates that the portion of the stabilizing layer nearest to the aqueous phase was the one most affected by the increase in the PEGylated phospholipid concentration. Table 1 provides the calculated values for the spectrum parameters. The mobility of the spin-probe and the time it takes for the spin-probe to make a full rotation is reflected in the τR parameter, which was changed the most, compared to the other parameters, by the variations in the PEGylated phospholipid content. A formulation with the most rigid stabilizing layer had the largest τR values, which, in this instance, was the formulation with 0.1% PEG2000-DSPE. It's interesting to note that the addition of PEGylated phospholipid had the opposite effect of strengthening the stabilizing layer. Further PEG2000-DSPE addition appeared to result in nanoemulsions with a less rigid stabilizing layer, possibly indicating that larger concentrations of the PEGylating agent lead to interface destabilization. The interactions between the curcumin, a symmetrical molecule with two aromatic ring systems and a bent conformation located in the stabilizing layer and the extra stabilizer are likely responsible for this.
The other two spin probes (12-DSA and 16-DSA) provide information about the stabilizing layer located closer to the oil core. Based on the data provided in Table 1 it can be inferred that the PEGylation mostly affects the stabilizing layer's areas closest to the aqueous interface, leaving the parts closer to the oil core largely not impacted.
CONCLUSION
This study demonstrates that one of the key elements in assessing how PEGylation affects the NE system is the active's localization. To pick the concentration of the PEGylated phospholipid that will offer the best surface coverage without compromising the integrity of the interface, additional considerations must be addressed in the event of an active situated in the stabilizing layer. In this instance, it may be hypothesized that the lowest PEG2000-DSPE concentration of 0.1%, CS21, will produce NEs that can slow down curcumin release the most compared to the other two formulations. Additionally, given that further addition of the PEGylated phospholipid causes the formation of less rigid stabilizing layer, further inquiries should be made to see the impact of these changes on the interactions with plasma proteins and biological fate of the droplets upon administration.
C3  - 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France
T1  - Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer
UR  - https://hdl.handle.net/21.15107/rcub_farfar_4572
ER  - 
@conference{
author = "Đoković, Jelena and Demisli, Sotiria and Papadimitriou, Vassiliki and Savić, Snežana",
year = "2023",
abstract = "INTRODUCTION
Nanoemulsions (NEs) offer a flexible platform for drug delivery via several administration routes. Rapid plasma clearance brought on by interactions with plasma proteins and the activation of the mononuclear phagocytic system is the greatest challenge NEs face after parenteral administration. PEGylation, or adding PEGylated phospholipids to the stabilizing layer of the NEs, is one method for ensuring that droplets circulate for a longer period of time. It is crucial to select the optimum concentration of the PEGylated in order to maintain the necessary physicochemical properties of NEs while providing appropriate surface coverage with the PEG chains. Curcumin is a model active that has been found to be localized in the stabilizing layer of NEs (1-3) and offers a wide range of potential health benefits, but due to its short plasma half-life, new strategies for enhancing bioavailability are required. The purpose of this study is to investigate the effects of various PEGylated phospholipid (PEG2000-DSPE) concentrations on the structural properties of NEs with an active placed in the stabilizing layer.
PREPARATION OF NANOEMULSIONS
All the NEs were prepared using the high pressure homogenization technique. The aqueous phase (glycerol, polysorbate 80, sodium oleate and highly purified water) was added to the oil phase (soybean oil, medium chain triglycerides, soybean lecithin, buthylhydroxytoluene, curcumin and 0.1%/0.3%/0.6% PEG2000-DSPE) and mixed at 11000rpm for 1 min on rotor stator homogenizer (IKA Ultra-Turrax T25 digital, IKA-Werke GmbH & Co. KG, Staufen, Germany), and then further processed at 800 bar for 10 discontinued cycles (EmulsiFlex-C3, Avestin Inc., Ottawa,ON, Canada) to obtain CS21, CS23 and CS26 formulations.
NANOEMULSION DROPLET SIZE
The droplet size was assessed through the dynamic light scattering method and presented as mean droplet size (Z-ave) and polydispersity index (PDI), after diluting the NEs 1:500 (v/v) in highly purified water.
ELECTRON PARAMAGNETIC RESONANCE (EPR) SPECTROSCOPY
For this study three amphiphilic fatty acid derivatives labeled at different positions of the aliphatic chain (5-DSA, 12-DSA and 16-DSA) were used to probe the dynamics of the membrane at different depths. Stock solutions of the spin probes were prepared in absolute ethanol at 1mM concentration. Subsequently, 15 µl of the stock solutions were evaporated and then incubated with 1 ml of the NE sample in final concentration of 0.015 mM. The resulting spectra was analyzed in terms of rotational correlation time (τR), order parameter (S) and isotropic hyperfine constant (αN).
RESULTS AND DISCUSION
All of the formulations had average droplet sizes between 95 and 103 nm and PDI values under 0.25, which indicated that they were suitable for parenteral administration.
The results of the EPR investigation showed that the stabilizing layer changed as the amount of PEGylated phospholipids increased, indicating that the PEGylation threshold has not yet been reached in the stabilizing layer.
The EPR research also showed that the 5-DSA spin probe's spectra were significantly affected by the addition of various PEGylated phospholipid concentrations (Figure 1). This indicates that the portion of the stabilizing layer nearest to the aqueous phase was the one most affected by the increase in the PEGylated phospholipid concentration. Table 1 provides the calculated values for the spectrum parameters. The mobility of the spin-probe and the time it takes for the spin-probe to make a full rotation is reflected in the τR parameter, which was changed the most, compared to the other parameters, by the variations in the PEGylated phospholipid content. A formulation with the most rigid stabilizing layer had the largest τR values, which, in this instance, was the formulation with 0.1% PEG2000-DSPE. It's interesting to note that the addition of PEGylated phospholipid had the opposite effect of strengthening the stabilizing layer. Further PEG2000-DSPE addition appeared to result in nanoemulsions with a less rigid stabilizing layer, possibly indicating that larger concentrations of the PEGylating agent lead to interface destabilization. The interactions between the curcumin, a symmetrical molecule with two aromatic ring systems and a bent conformation located in the stabilizing layer and the extra stabilizer are likely responsible for this.
The other two spin probes (12-DSA and 16-DSA) provide information about the stabilizing layer located closer to the oil core. Based on the data provided in Table 1 it can be inferred that the PEGylation mostly affects the stabilizing layer's areas closest to the aqueous interface, leaving the parts closer to the oil core largely not impacted.
CONCLUSION
This study demonstrates that one of the key elements in assessing how PEGylation affects the NE system is the active's localization. To pick the concentration of the PEGylated phospholipid that will offer the best surface coverage without compromising the integrity of the interface, additional considerations must be addressed in the event of an active situated in the stabilizing layer. In this instance, it may be hypothesized that the lowest PEG2000-DSPE concentration of 0.1%, CS21, will produce NEs that can slow down curcumin release the most compared to the other two formulations. Additionally, given that further addition of the PEGylated phospholipid causes the formation of less rigid stabilizing layer, further inquiries should be made to see the impact of these changes on the interactions with plasma proteins and biological fate of the droplets upon administration.",
journal = "4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France",
title = "Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer",
url = "https://hdl.handle.net/21.15107/rcub_farfar_4572"
}
Đoković, J., Demisli, S., Papadimitriou, V.,& Savić, S.. (2023). Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France.
https://hdl.handle.net/21.15107/rcub_farfar_4572
Đoković J, Demisli S, Papadimitriou V, Savić S. Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer. in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France. 2023;.
https://hdl.handle.net/21.15107/rcub_farfar_4572 .
Đoković, Jelena, Demisli, Sotiria, Papadimitriou, Vassiliki, Savić, Snežana, "Structural analysis of PEGylated nanoemulsions using EPR spectroscopy – the impact of an active compound incorporated in stabilizing layer" in 4th European Conference on Pharmaceutics, 20 - 21 March 2023, Marseille, France (2023),
https://hdl.handle.net/21.15107/rcub_farfar_4572 .

Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment

Vukašinović, Mila; Savić, Sanela; Cekić, Nebojša; Ilić, Tanja; Pantelić, Ivana; Savić, Snežana

(MDPI, 2023)

TY  - JOUR
AU  - Vukašinović, Mila
AU  - Savić, Sanela
AU  - Cekić, Nebojša
AU  - Ilić, Tanja
AU  - Pantelić, Ivana
AU  - Savić, Snežana
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4517
AB  - Since natural-origin, sustainable ingredients are preferred by modern consumers, novel emulsifiers and emollients keep entering the market. This study hypothesizes that a combination of in silico, instrumental tools and simplified sensory studies could be used to efficiently characterize emulsions in a shorter timeframe. A total of 22 rather simple o/w emulsions were prepared by a time/energy-saving emulsification process. A natural mixed emulsifier (Lauryl Glucoside/Myristyl Glucoside/Polyglyceryl-6 Laurate) and two emollients (both with INCI name C15–19 Alkane) were used. The performed D-optimal experimental design within the response surface method (RSM) significantly narrowed down the number of samples about to enter the stage of texture, friction and sensory studies to the samples comprising 30% of a respective Emogreen emollient and 2% or 3% of the emulsifier. The sample comprising 2% emulsifier/30% Emogreen® L15 showed significantly higher firmness (42.12 mN) when compared to the one with 2% emulsifier/30% Emogreen® L19 (33.62 mN), which was somewhat unexpected considering the emollients’ inherent viscosity values (4.5 mPa·s for L15 and 9 mPa·s for L19). The sample with 2% emulsifier/30% Emogreen® L19 managed to maintain the lowest friction, while the one with 3% emulsifier/30% Emogreen® L19 released its full lubricating potential in the second part of the measurement (30–60 s). The obtained results revealed the strengths and weaknesses of each formulation, narrowing down their possible applications in the early development stage.
PB  - MDPI
T2  - Pharmaceutics
T1  - Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment
VL  - 15
IS  - 2
DO  - 10.3390/pharmaceutics15020486
ER  - 
@article{
author = "Vukašinović, Mila and Savić, Sanela and Cekić, Nebojša and Ilić, Tanja and Pantelić, Ivana and Savić, Snežana",
year = "2023",
abstract = "Since natural-origin, sustainable ingredients are preferred by modern consumers, novel emulsifiers and emollients keep entering the market. This study hypothesizes that a combination of in silico, instrumental tools and simplified sensory studies could be used to efficiently characterize emulsions in a shorter timeframe. A total of 22 rather simple o/w emulsions were prepared by a time/energy-saving emulsification process. A natural mixed emulsifier (Lauryl Glucoside/Myristyl Glucoside/Polyglyceryl-6 Laurate) and two emollients (both with INCI name C15–19 Alkane) were used. The performed D-optimal experimental design within the response surface method (RSM) significantly narrowed down the number of samples about to enter the stage of texture, friction and sensory studies to the samples comprising 30% of a respective Emogreen emollient and 2% or 3% of the emulsifier. The sample comprising 2% emulsifier/30% Emogreen® L15 showed significantly higher firmness (42.12 mN) when compared to the one with 2% emulsifier/30% Emogreen® L19 (33.62 mN), which was somewhat unexpected considering the emollients’ inherent viscosity values (4.5 mPa·s for L15 and 9 mPa·s for L19). The sample with 2% emulsifier/30% Emogreen® L19 managed to maintain the lowest friction, while the one with 3% emulsifier/30% Emogreen® L19 released its full lubricating potential in the second part of the measurement (30–60 s). The obtained results revealed the strengths and weaknesses of each formulation, narrowing down their possible applications in the early development stage.",
publisher = "MDPI",
journal = "Pharmaceutics",
title = "Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment",
volume = "15",
number = "2",
doi = "10.3390/pharmaceutics15020486"
}
Vukašinović, M., Savić, S., Cekić, N., Ilić, T., Pantelić, I.,& Savić, S.. (2023). Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment. in Pharmaceutics
MDPI., 15(2).
https://doi.org/10.3390/pharmaceutics15020486
Vukašinović M, Savić S, Cekić N, Ilić T, Pantelić I, Savić S. Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment. in Pharmaceutics. 2023;15(2).
doi:10.3390/pharmaceutics15020486 .
Vukašinović, Mila, Savić, Sanela, Cekić, Nebojša, Ilić, Tanja, Pantelić, Ivana, Savić, Snežana, "Efficient Development of Green Emulsifier/Emollient-Based Emulsion Vehicles: From RSM Optimal Experimental Design to Abridged In Vivo Assessment" in Pharmaceutics, 15, no. 2 (2023),
https://doi.org/10.3390/pharmaceutics15020486 . .
3
3

Parenteral Lipid-Based Nanoparticles for CNS Disorders: Integrating Various Facets of Preclinical Evaluation towards More Effective Clinical Translation

Ilić, Tanja; Đoković, Jelena; Nikolić, Ines; Mitrović, Jelena; Pantelić, Ivana; Savić, Snežana; Savić, Miroslav

(MDPI, 2023)

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 . .
5
2

High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances

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

(Elsevier B.V., 2023)

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 . .
3
2

Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study

Demisli, Sotiria; Galani, Eleni; Goulielmaki, Maria; Kyrilis, Fotios; Ilić, Tanja; Hamdi, Farzad; Crevar, Milkica; Kastritis, Panagiotis; Pletsa, Vasiliki; Nallet, Frédéric; Savić, Snežana; Xenakis, Aristotelis; Papadimitriou, Vassiliki

(Academic Press Inc., 2023)

TY  - JOUR
AU  - Demisli, Sotiria
AU  - Galani, Eleni
AU  - Goulielmaki, Maria
AU  - Kyrilis, Fotios
AU  - Ilić, Tanja
AU  - Hamdi, Farzad
AU  - Crevar, Milkica
AU  - Kastritis, Panagiotis
AU  - Pletsa, Vasiliki
AU  - Nallet, Frédéric
AU  - Savić, Snežana
AU  - Xenakis, Aristotelis
AU  - Papadimitriou, Vassiliki
PY  - 2023
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4384
AB  - Hypothesis: Lipophilic cannabidiol can be solubilized in oil-in water nanoemulsions, which can then be impregnated into chitosan hydrogels forming another colloidal system that will facilitate cannabidiol's release. The delivery from both systems was compared, alongside structural and biological studies, to clarify the effect of the two carriers' structure on the release and toxicity of the systems. Experiments: Oil-in-water nanoemulsions (NEs) and the respective nanoemulsion-filled chitosan hydrogels (NE/HGs) were formulated as carriers of cannabidiol (CBD). Size, polydispersity and stability of the NEs were evaluated and then membrane dynamics, shape and structure of both systems were investigated with EPR spin probing, SAXS and microscopy. Biocompatibility of the colloidal delivery systems was evaluated through cytotoxicity tests over normal human skin fibroblasts. An ex vivo permeation protocol using porcine ear skin was implemented to assess the release of CBD and its penetration through the skin. Findings: Incorporation of the NEs in chitosan hydrogels does not significantly affect their structural properties as evidenced through SAXS, EPR and confocal microscopy. These findings indicate the successful development of a novel nanocarrier that preserves the NE structure with the CBD remaining encapsulated in the oil core while providing new rheological properties advantageous over NEs. Moreover, NE/HGs proved to be more efficient as a carrier for the release of CBD. Cell viability assessment revealed high biocompatibility of the proposed colloids.
PB  - Academic Press Inc.
T2  - Journal of Colloid and Interface Science
T1  - Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study
VL  - 634
SP  - 300
EP  - 313
DO  - 10.1016/j.jcis.2022.12.036
ER  - 
@article{
author = "Demisli, Sotiria and Galani, Eleni and Goulielmaki, Maria and Kyrilis, Fotios and Ilić, Tanja and Hamdi, Farzad and Crevar, Milkica and Kastritis, Panagiotis and Pletsa, Vasiliki and Nallet, Frédéric and Savić, Snežana and Xenakis, Aristotelis and Papadimitriou, Vassiliki",
year = "2023",
abstract = "Hypothesis: Lipophilic cannabidiol can be solubilized in oil-in water nanoemulsions, which can then be impregnated into chitosan hydrogels forming another colloidal system that will facilitate cannabidiol's release. The delivery from both systems was compared, alongside structural and biological studies, to clarify the effect of the two carriers' structure on the release and toxicity of the systems. Experiments: Oil-in-water nanoemulsions (NEs) and the respective nanoemulsion-filled chitosan hydrogels (NE/HGs) were formulated as carriers of cannabidiol (CBD). Size, polydispersity and stability of the NEs were evaluated and then membrane dynamics, shape and structure of both systems were investigated with EPR spin probing, SAXS and microscopy. Biocompatibility of the colloidal delivery systems was evaluated through cytotoxicity tests over normal human skin fibroblasts. An ex vivo permeation protocol using porcine ear skin was implemented to assess the release of CBD and its penetration through the skin. Findings: Incorporation of the NEs in chitosan hydrogels does not significantly affect their structural properties as evidenced through SAXS, EPR and confocal microscopy. These findings indicate the successful development of a novel nanocarrier that preserves the NE structure with the CBD remaining encapsulated in the oil core while providing new rheological properties advantageous over NEs. Moreover, NE/HGs proved to be more efficient as a carrier for the release of CBD. Cell viability assessment revealed high biocompatibility of the proposed colloids.",
publisher = "Academic Press Inc.",
journal = "Journal of Colloid and Interface Science",
title = "Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study",
volume = "634",
pages = "300-313",
doi = "10.1016/j.jcis.2022.12.036"
}
Demisli, S., Galani, E., Goulielmaki, M., Kyrilis, F., Ilić, T., Hamdi, F., Crevar, M., Kastritis, P., Pletsa, V., Nallet, F., Savić, S., Xenakis, A.,& Papadimitriou, V.. (2023). Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study. in Journal of Colloid and Interface Science
Academic Press Inc.., 634, 300-313.
https://doi.org/10.1016/j.jcis.2022.12.036
Demisli S, Galani E, Goulielmaki M, Kyrilis F, Ilić T, Hamdi F, Crevar M, Kastritis P, Pletsa V, Nallet F, Savić S, Xenakis A, Papadimitriou V. Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study. in Journal of Colloid and Interface Science. 2023;634:300-313.
doi:10.1016/j.jcis.2022.12.036 .
Demisli, Sotiria, Galani, Eleni, Goulielmaki, Maria, Kyrilis, Fotios, Ilić, Tanja, Hamdi, Farzad, Crevar, Milkica, Kastritis, Panagiotis, Pletsa, Vasiliki, Nallet, Frédéric, Savić, Snežana, Xenakis, Aristotelis, Papadimitriou, Vassiliki, "Encapsulation of cannabidiol in oil-in-water nanoemulsions and nanoemulsion-filled hydrogels: A structure and biological assessment study" in Journal of Colloid and Interface Science, 634 (2023):300-313,
https://doi.org/10.1016/j.jcis.2022.12.036 . .
9
8

Estimation of endotoxin level in nanocrystal dispersion of DK-I-56-1 intended for parenteral administration

Mitrović, Jelena; Ilić, Tanja; Jančić, Ivan; Bufan, Biljana; Savić, Miroslav; Savić, Snežana

(2023)

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 .

Preformulation and development of preliminary nanoemulsion carrier for patent protected compound GL-II-73

Đoković, Jelena; Marković, Bojan; Sharmin, Dishary; Cook, James M.; Savić, Miroslav; Savić, Snežana

(Macedonian Pharmaceutical Association, 2023)

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 . .

Searching for the best way to incorporate the proprietary compound GL-II -73 into the nanoemulsion carrier for prospective parenteral application

Đoković, Jelena; Marković, Bojan; Sharmin, Dishary; Cook, James M.; Savić, Miroslav; Savić, Snežana

(International Association of Physical Chemists, 2023)

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 .

Biocompatible nanoemulsions as a tool for preclinical testing of CW-02-79, a pyrazoloquinolinone modulator of sigma-2 receptors: preformulation and formulation studies

Ilić, Tanja; Stanković, Tijana; Mitrović, Jelena; Pantelić, Ivana; Dobričić, Vladimir; Cook, James M.; Savić, Miroslav; Savić, Snežana

(2023)

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 .