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The use of cyclodextrins in the development of therapeutic delivery systems for antitumor drugs
No full textOnkološke bolesti su drugi uzrok smrtnosti u svijetu, a broj oboljelih raste. Terapija tumora uništava i zdrave, normalne stanice, osobito one koje se brzo dijele što dovodi do brojnih nuspojava. Citostatici su većinom slabo topljivi u vodi pa ih je teško formulirati, a pomoćne tvari koje se koriste u tu svrhu često uzrokuju dodatne nuspojave. Ciklodekstrini omogućuju povećanje topljivosti citostatika bez izazivanja novih nuspojava. To su biokomptibilni ciklički oligosaharidi izgrađeni iz α-1,4-D-glukopiranoznih jedinica. Strukture su šupljeg krnjeg stošca čija je unutrašnjost hidrofobna, a vanjština hidrofilna. Uklapanjem slabo topljivih lijekova u svoju šupljinu povećavaju im topljivost, maskiraju okus i miris lijeka, sprječavaju iritaciju gastrointestinalnog trakta ili sprječavaju interakcije s drugim lijekovima.
CD se primjenjuju kao solubilizatori pri čemu povećavaju bioraspoloživost citostatika, a smanjuju im toksičnost. Klinička ispitivanja melfalana uklopljenog u CD pokazala su objektivnu stopu odgovora od 95%, a teške nuspojave koje se povezuju s konvecionalnom formulacijom melfalana izbjegnute su. CD polimeri s kamptotecinom stvaraju CRLX101 nanočestice koje imaju do 1000 puta veću topljivost, a zbog EPR učinka smanjena je sistemska toksičnost. Klinička ispitivanja CRLX101 pokazala su stopu kliničkog benefita od 68% za monoterapiju i čak 95% za kombinaciju s bevacizumabom. Također, CD se koriste i kako bi se spriječila inkompatibilnost alkalne otopine 5-FU s kiselim leukovorinom. Naime, CD omogućavaju topljivost 5-FU pri fiziološkoj pH vrijednosti pa se, osim reakcije neutralizacije, izbjegava i kardiotoksičnost povezana s primjenom alkalne otopine. Razvijaju se i istražuju napredni sustavi dostave citostatika u tumorske stanice pri čemu se CD modificiraju za ciljanu dostavu ili lakše uklapanje gena.Oncologycal diseases are the second leading cause of death worldwide, with a growing number of patients. Tumor therapy also destroys healthy normal cells, especially ones that divide quickly, which leads to numerous adverse effects, Cytostatics are usually poorly soluble in water which makes them hard to formulate and other excipients often cause additional adverse effects. Cyclodextrins can be used to improve drug solubility without aggravating side effects. Cyclodextrins are biocompatible cyclic oligosaccharides built from α-1,4-D-glucopyranose units. Spatially, they form an inverted hollow truncated cone, the interior of which is hydrophobic, and the exterior is hydrophilic. By fitting poorly soluble drugs into their cavity, they increase their solubility, mask the taste and smell of the drug, prevent irritation of the gastrointestinal tract or prevent interactions with other drugs.
CDs are used as solubilizers, increasing the bioavailability of cytostatics and reducing their toxicity. Clinical trials of melphalan incorporated into CD have shown an objective response rate of 95%, and the severe side effects associated with conventional melphalan formulation have been avoided. CD polymers create CRLX101 nanoparticles with camptothecin that have up to 1000 times greater solubility, and due to the EPR effect, systemic toxicity is reduced. Clinical trials of CRLX101 showed a clinical benefit rate of 68% for monotherapy and even 95% for the combination with bevacizumab. Also, CDs are used to prevent the incompatibility of the alkaline solution of 5-FU with the acidic leucovorin. Namely, CDs enable the solubility of 5-FU at a physiological pH value, so, in addition to the neutralization reaction, the cardiotoxicity associated with the use of an alkaline solution is also avoided. Advanced delivery systems for cytostatics to tumor cells are being developed and investigated, whereby CDs are modified for targeted delivery or easier gene insertion
The significance of determining complement component C1q concentration in patients with systemic lupus erythematosus
No full textSustav komplementa čini više od 30 proteina prisutnih u plazmi i na staničnim membranama, čije funkcije
uključuju moduliranje imunosnog odgovora, liziranje stanica patogena te uklanjanje staničnog otpada i
imunokompleksa. Komponenta komplementa C1q prepoznaje imunokomplekse nastale kao dio imunosnog
odgovora te započinje klasičan put aktivacije sustava komplementa. Sistemski eritemski lupus (SLE) je
multisistemska autoimuna bolest koju karakterizira nastanak te odlaganje imunokompleksa u različitim organima.
Najteža komplikacija SLE koja nastaje nakupljanjem imunokompleksa u bubrezima naziva se lupusni nefritis. Cilj
ovog istraživanja bio je utvrditi važnost određivanja koncentracije komponente komplementa C1q kao
potencijalnog biomarkera u serumu pacijenata sa SLE te povezanost s komponentama komplementa C3 i C4 koje
su dio redovne obrade i praćenja bolesnika. Istraživanje je provedeno u 101 ispitaniku, od kojih 33 zdravih i 68
oboljelih od SLE. Na temelju izmjerenih koncentracija komponente komplementa C1q u serumu zdravih
ispitanika, izračunat je referentni interval koji iznosi 20,7 - 33,4 mg/dL, te se razlikovao od referentnog intervala
predloženim od proizvođača (15,7 - 30,6 mg/dL). Usporedbom vrijednosti koncetracija C1q dobivenih u uzorcima
zdravih i bolesnik ispitanika, utvrđena je statisitički značajno niža vrijednost koncetracije C1q u serumu bolesnih
ispitanika (P<0,001). U skupini bolesnih ispitanika utvrđena je statistički značajna korelacija između koncentracije
komponente komplementa C1q i koncetnracije komponenata komplementa C3 (r=0,450; P=<0,001) i C4
(ρ=0,460; P=<0,001). Dobiveni rezultati podudarni su s dosadašnjim spoznajama o SLE i literaturnim navodima.
Utvrđene značajne razlike i trendovi pokazuju kako se komponenta komplementa C1q može smatrati potencijalnim
biomarkerom, ali je istraživanje potrebno nadopuniti upotrebom različitih analitičkih sustava te većim brojem
ispitanika.The complement system consists of more than 30 proteins present in plasma and on cell membranes, whose
functions include modulation of the immune response, lysis of pathogen cells, and removal of cellular debris and
immune complexes. The complement component C1q recognizes immune complexes formed as part of the
immune response and initiates the classical pathway of complement activation. Systemic lupus erythematosus
(SLE) is a multisystem autoimmune disease characterized by the formation and deposition of immune complexes
in various organs. The most severe complication of SLE resulting from the accumulation of immune complexes in
the kidneys is termed lupus nephritis. The aim of this study was to determine the significance of measuring the
concentration of complement component C1q as a potential biomarker in the serum of patients with SLE, as well
as its correlation with complement components C3 and C4, which are part of routine patient evaluation and
monitoring. The study was conducted on 101 participants, including 33 healthy individuals and 68 patients with
SLE. Based on the measured concentrations of complement component C1q in the serum of healthy subjects, a
reference interval was calculated to be 20.7 - 33.4 mg/dL, which differed from the reference interval proposed by
the manufacturer (15.7 - 30.6 mg/dL). Comparison of C1q concentrations obtained from healthy and patient
samples revealed a statistically significant lower concentration of C1q in the serum of patients (P<0.001). In the
patient group, a statistically significant correlation was found between the concentration of complement
component C1q and the concentrations of complement components C3 (r=0.450; P<0.001) and C4 (ρ=0.460;
P<0.001). The obtained results are consistent with existing knowledge regarding SLE and literature references.
The identified significant differences and trends suggest that complement component C1q may be considered a
potential biomarker; however, further research is necessary, incorporating various analytical systems and a larger
number of participants
Comparison of LC-MS and nano-LC-MS methods for the analysis of N-glycosylation of alpha-1-acid glycoprotein
No full textGlikozilacija proteina složena je posttranslacijska modifikacija koja ima mnoge biološke uloge u
organizmu. Abnormalnosti u glikozilaciji primijećene su u nekoliko različitih bolesti te su razlike u
glikozilaciji proteina povezane s osjetljivošću na bolest, tijekom bolesti i odgovorom na terapiju.
Alfa-1 kiseli glikoprotein (AGP) jedan je od glavnih proteina akutne faze kod ljudi. Ima 5 Nglikozilacijskih mjesta, a na ta mjesta kovalentno se vežu različiti tipovi glikana. Nedavno je Nglikanski profil AGP-a analiziran nano-LC-MS metodom pri čemu je pokazano da pojedinci s većim
rizikom od dijabetesa imaju povećano grananje AGP N-glikana i nižu sijalinizaciju na nekim
glikozilacijskim mjestima, te bi AGP N-glikanski profil mogao pomoći u razlikovanju pojedinaca
koji su u riziku od razvoja dijabetesa tipa 2.
U ovom radu cilj je bio ispitati može li se glikozilacija AGP-a jednako učinkovito analizirati
korištenjem klasične UHPLC-MS metode koja je jednostavnija, robusnija i lakša za korištenje od
nano-LC-MS-a. AGP je najprije obogaćen iz uzorka precipitacijom „seromukoidne“ frakcije, nakon
čega je uzorak prošao kroz redukciju, alkilaciju i razgradnju enzimom tripsinom. Zatim su
glikopeptidi AGP-a obogaćeni pomoću ekstrakcije na čvrstoj fazi pomoću HILIC zrnaca. Potom su
dobiveni AGP glikopeptidi analizirani pomoću nano-LC-MS-a, te UHPLC-MS-a. Samo su određeni
glikopeptidi prošli parametre kontrole kvalitete (QC), 53 analita u UHPLC-MS-u, a 83 analita u
nano-LC-MS-u. Glikopeptidi koji se preklapaju u obje metode izdvojeni su i normalizirani na
ukupnu integriranu površinu po pojedinom glikozilacijskom mjestu kojem pripadaju. Kako bi se
usporedila ponovljivost između dviju metoda, izračunat je koeficijent varijacije, a kako bi se utvrdila
korelacija, koristio se koeficijent korelacije (r) te je izračunat i koeficijent determinacije (R2).
Pokazano je da obje metode imaju usporedivu ponovljivost, te je vidljiva dobra korelacija u
relativnom intenzitetu većine glikopeptida. U slučaju određenih glikopeptida, pogotovo u onih s
nižim udjelima, ipak postoje razlike u rezultatima. To se može pripisati manjoj osjetljivosti UHPLCMS metode. Unatoč tome, UHPLC-MS je brža, jednostavnija i prikladnija za analizu velikog broja
uzoraka, što je čini idealnom za visoko-propusne analize. Sveukupno, obje metode imaju svoje
mjesto u analizi N-glikozilacije AGP-a, a izbor metode ovisi o specifičnim zahtjevima analize.Glycosylation of proteins is a complex post-translational modification with numerous biological
roles in the organism. Abnormalities in glycosylation have been observed in various diseases, and
differences in protein glycosylation are linked to disease susceptibility, disease progression, and
therapeutic response. Alpha-1-acid glycoprotein (AGP) is one of the main acute-phase proteins in
humans. It has 5 N-glycosylation sites, to which various types of glycans are covalently attached.
Recently, the N-glycan profile of AGP was analyzed using the nano-LC-MS method, showing that
individuals at higher risk of diabetes have increased branching of AGP N-glycans and reduced
sialylation at certain glycosylation sites, suggesting that the AGP N-glycan profile could help
distinguish individuals at risk of developing type 2 diabetes.
The aim of this study was to determine whether AGP glycosylation can be analyzed just as
effectively using the conventional UHPLC method, which is simpler, more robust, and easier to use
than nano-LC-MS. AGP was first enriched from the sample by precipitating the “seromucoid”
fraction, followed by reduction, alkylation, and digestion with trypsin. AGP glycopeptides were then
enriched using solid-phase extraction with HILIC beads. The AGP glycopeptides were then analyzed
by both nano-LC-MS and UHPLC-MS. Only certain glycopeptides met the quality control (QC)
criteria, with 53 analytes in UHPLC-MS and 83 analytes in nano-LC-MS. Overlapping
glycopeptides from both methods were selected and normalized to the total integrated area of their
respective glycosylation sites. To compare the repeatability of the two methods, the coefficient of
variation was calculated, and the correlation was assessed using the correlation coefficient (r). The
coefficient of determination (R²) was also calculated.
It was shown that both methods have comparable repeatability, with good correlation in the relative
intensity of most glycopeptides. However, for certain glycopeptides, especially those with lower
relative intensity, differences in results were observed. This can be attributed to the lower sensitivity
of the UHPLC-MS method. Despite this, UHPLC-MS is faster, simpler, and more suitable for
analyzing a large number of samples, making it ideal for high-throughput analyses. Overall, both
methods have their place in the analysis of AGP N-glycosylation, and the choice of method depends
on the specific requirements of the analysis
Drug therapy problems related to benzodiazepines and Z-drugs in primary healthcare
No full textCilj istraživanja: Utvrditi i definirati najčešće terapijske probleme u pacijenata s barem jednim lijekom iz skupine
benzodiazepina i z-lijekova na razini primarne zdravstvene zaštite u Republici Hrvatskoj.
Materijali i metode: Provedeno je prospektivno, intervencijsko istraživanje u FTS unutar DZZC gdje se provodila usluga
upravljanja farmakoterapijom u razdoblju od siječnja 2018. do travnja 2024. Prikupljeni su sociodemografski i antropometrijski
podatci o pacijentu, njegovim bolestima i lijekovima te su identificirani terapijski problemi i izrađen je plan skrbi s predloženim
intervencijama. Kriterij za izbor sudionika bio je ustanovljen terapijski problem povezan s primjenom benzodiazepina ili z-lijeka.
Podatci su analizirani metodom deskriptivne statistike pomoću Microsoft Excel programa.
Rezultati: U istraživanje je bilo uključeno 118 pacijenata, od kojih su 77,1 % bile žene, prosječne dobi 71,2 ± 11 godina.
Prosječno su pacijenti imali 10,2±6,0 komorbiditeta i 11,2±5,1 lijekova. Identificirano je ukupno 1042 terapijskih problema
(8,83±3,0 po pacijentu), od čega se 14,5 % odnosilo na benzodiazepine i z-lijekove (prosječno 1,28 po pacijentu). Najčešći
terapijski problemi bili su previsoka doza (53,2 % za benzodiazepine, 55,6 % za z-lijekove), nepotrebna terapija (23,4 % i 11,1
%) te nuspojava lijeka (13,7 % i 14,8 %). Najčešće predložena intervencija bila je prekid terapije (45 % za benzodiazepine i 40,7
% za z-lijekove). Ukupno je 52,4 % terapijskih problema povezanih s primjenom benzodiazepina i 37,0 % povezanih s primjenom z-lijekova bilo riješeno.
Zaključak: Rezultati ovog istraživanja pokazuju da ukoliko farmaceut u radu koristi jasno definiran i standardiziran proces rada
ljekarničke prakse, u suradnji sa liječnicima obiteljske medicine i pacijentima, može riješiti velik broj terapijskih problema
povezanih s neracionalnom primjenom benzodiazepina i z- lijekova.Objective: To identify and define the most common drug therapy problems in patients using at least one drug from the
benzodiazepine and z-drug groups at the primary healthcare level in the Republic of Croatia.
Materials and Methods: A prospective, interventional study was conducted within the FTS at the DZZC, where a
comprehensive medication management service was provided from January 2018 to April 2024. Sociodemographic and
anthropometric data about the patients, their diseases, and medications were collected, and drug therapy problems were identified.
A care plan with proposed interventions was developed. The inclusion criterion for participants was the presence of a drug
therapy problem related to the use of benzodiazepines or z-drugs. The data were analyzed using descriptive statistics with
Microsoft Excel.
Results: A total of 118 patients were included in the study, of which 77.1% were women, with an average age of 71.2 ± 11 years. On average, patients had 10.2 ± 6.0 comorbidities and 11.2 ± 5.1 medications. A total of 1042 drug therapy problems were identified (8.83 ± 3.0 per patient), of which 14.5% were related to benzodiazepines and z-drugs (an average of 1.28 per patient). The most common drug therapy problems were excessive dosage (53.2% for benzodiazepines, 55.6% for z-drugs), unnecessary therapy (23.4% and 11.1%), and drug side effects (13.7% and 14.8%). The most frequently proposed intervention was discontinuing the therapy (45% for benzodiazepines and 40.7% for z-drugs). In total, 52.4% of drug therapy problems related to the use of benzodiazepines and 37.0% related to z-drugs were resolved.
Conclusion: The results of this study show that when a pharmacist employs a clearly defined and standardized working process in pharmacy practice, in collaboration with family medicine doctors and patients, a large number of drug therapy problems related to the irrational use of benzodiazepines and z-drugs can be resolved
Ultrasound-assisted extraction of polyphenolics of Plantago major L.
No full textVeliki ili širokolisni trputac (Plantago major L., Plantaginaceae) je široko rasprostranjena biljka, koja je
poznata u narodnoj medicini već dugi niz godina. Sadrži brojne biološki aktivne spojeve, a to su:
flavonoidi, alkaloidi, terpenoidi, iridoidi, vitamini, polisaharidi, masne i fenolne kiseline. Zahvaljujući svom
kemijskom sastavu posjedu brojne biološke učinke. Primjerice, pomaže u zacjeljivanju rana, koristan je kod
dijareje, pomaže u prevenciji virusnih i bakterijskih infekcija, te snižava razinu glukoze i kolesterola u krvi.
Polifenoli djeluju protuupalno, antioksidacijski, te smanjuju rizik od nekih kroničnih bolesti, npr.
kardiovaskularnih. U ovom radu je za ekstrakciju polifenola iz P.major korištena brza i učinkovita metoda
ekstrakcije, ultrazvučna ekstrakcija. Spektrofotometrijski je određena količina ukupnih polifenola,
flavonoida i fenolnih kiselina. Koncentracija verbaskozida određena je korištenjem HPLC-a. Usporedbom
koncentracija spojeva i uvjeta pripreme ekstrakta ustanovilo se koji su parametri uvjetovali najefikasniju
ekstrakciju pojedinih polifenola. Najveći sadržaj ukupnih polifenola imao je ekstrakt koji sadrži 70 % GL, 5
% mliječne kiseline, a ekstrahirao se 30 min pri temperaturi od 70°C i snazi ultrazvuka od 144 W.
Flavonoidi su se najbolje ekstrahirali sa 70 % GL, bez mliječne kiseline, a ekstrakcija je trajala 30 min pri
70 °C i snazi ultrazvuka od 720 W. Fenolnim kiselinama je odgovarao medij s 10 % GL i 0,3 mmol HP-β-
CD, a ekstrakcija je trajala 30 min pri 20 °C i snazi ultrazvuka od 144 W. Ekstrakcija verbaskozida je
najučinkovitije provedena s 10 % GL, 5% mliječne kiseline i 0,3 mmol HP-β-CD, a ekstrakcijsko vrijeme je
bilo 10 min na 20 °C i pri snazi ultrazvuka od 144 W. Potrebno je statistički obraditi rezultate kako bi se
preciznije odredilo koliko pojedini parametri utječu na ekstrakciju svih navedenih polifenolnih sastavnica.Greater or broadleaf plantain (Plantago major L., Plantaginaceae) is a widespread plant, known in folk medicine
for many years. It contains numerous biologically active compounds including flavonoids, alkaloids, terpenoids,
iridoids, vitamins, polysaccharides, fatty and phenolic acids. Due to its chemical constituents, it has numerous
biological effects. For example, it promotes wound healing, prevents viral and bacterial infections, it is useful in
treatment of diarrhea, and lowers the level of glucose and cholesterol in the blood. Polyphenols act as antioxidants,
anti-inflammatory and reduce the risk of some chronic diseases, eg cardiovascular. In this work, a fast and
efficient extraction method, ultrasonic extraction, was used for the extraction of polyphenols from P. major. The
amount of total polyphenols, flavonoids and phenolic acids was determined spectrophotometrically. Verbascoside
concentration was determined using HPLC. By comparing the concentrations of the target compounds and the
conditions in which extracts were prepared, the parameters necesary for the most efficient extraction of individual
polyphenols were established. The extract with the highest content of total polyphenols was prepared using 70 %
GL, 5 % lactic acid and was extracted for 30 min at a temperature of 70 °C with an ultrasound power of 144 W.
Flavonoids were best extracted using 70 % GL, without lactic acid, and the extraction lasted 30 min at 70 °C with
an ultrasound power of 720 W. The most suitable medium for phenolic acid was with 10 % GL, 0.3 mmol HP-β-
CD, and the extraction lasted 30 min at 20 °C with an ultrasound power of 144 W. The extract with 10 % GL, 5%
lactic acid, 0.3 mmol HP-β-CD showed the highest verbascoside extraction, and the extraction time was 10 min at
20 °C and with an ultrasound power of 144 W. It is still necessary to statistically process the results in order to
precisely determine how certain parameters affect the extraction of all the mentioned polyphenol components
Utjecaj koronavirusa na crijevnu mikrobiotu
No full textCilj istraživanja: Ravnoteža crijevne mikrobiote od velike je važnosti za očuvanje zdravlja ljudi. Stanje kada je ravnoteža među komponentama mikrobiote narušena, naziva se disbioza. Godine 2019. u kineskom gradu Wuhan izbijala je bolest teškog akutnog respiratornog sindroma nazvana COVID-19, koja je uzrokovana novim koronavirusom SARS-CoV-2. Osim infekcije respiratornog sustava, SARS-CoV-2 također inficira gastrointestinalni trakt, gdje se intenzivno razmnožava. Cilj ovog rada je pružiti uvid u promjene sastava crijevne mikrobiote kod pacijenata s COVID-19 i kod osoba koje su se oporavile od COVID-19, kao i uvid u terapijske intervencije koje utječu na promjenu sastava crijevne mikrobiote u pacijenata s COVID-19.
Metode: Za izradu ovog preglednog rada pretraživana je dostupna znanstvena literatura na temu crijevne mikrobiote, COVID-19, analize crijevne mikrobiote u COVID-19 pacijenata te utjecaja probiotika i prehrane na crijevnu mikrobiotu kod COVID-19 pacijenata.
Rezultati: Sastav crijevne mikrobiote COVID-19 pacijenata te osoba koje su se oporavile od COVID-19 pokazuje značajnu disbiozu u usporedbi sa zdravim kontrolama. Dokazano je smanjenje mikrobne raznolikosti i mikrobnog bogatstva, kao i promjene u sastavu crijevne mikrobiote u obliku gubitka korisnih bakterija i povećanja brojnosti patogenih bakterija. Disbioza crijeva usko je povezana s gastrointestinalnim simptomima i težinom bolesti COVID-19.
Zaključak: Za utvrđivanje odražavaju li te promjene u sastavu crijevne mikrobiote uzrok ili posljedicu COVID-19, potrebno je provesti dodatna istraživanja. Terapijska intervencija na mikrobiotu putem probiotika, prebiotika i fekalne transplantacije je jedna od obećavajućih strategija za liječenje COVID-19 u budućnosti, ali neophodna su daljnja istraživanjaObjectives: The intestinal microbiota balance is of great importance for maintaining human health. The condition when the balance in the microbial community is disturbed is called dysbiosis. In 2019, a severe acute respiratory syndrome disease called COVID-19 broke out in the Chinese city of Wuhan. It is an infectious disease caused by the new coronavirus SARS-CoV-2. In addition to infection of the respiratory system, SARS-CoV-2 also infects the gastrointestinal tract, where it multiplies intensively. The aim of this work is to provide insight into changes in the intestinal microbiota compostion in patients with COVID-19 and in people who have recovered from COVID-19, as well as insight into therapeutic interventions that affect changes in the intestinal microbiota composition in patients with COVID-19.
Methods: The methods included a review of available scientific literature on the topic of intestinal microbiota, COVID-19, analysis of intestinal microbiota in COVID-19 patients, and the impact of probiotics and nutrition on intestinal microbiota in COVID-19 patients.
Results: The composition of the intestinal microbiota of COVID-19 patients and persons who have recovered from COVID-19 shows significant dysbiosis compared to healthy controls. A decrease in microbial diversity and richness, as well as changes in the composition of the intestinal microbiota in the form of a loss of beneficial bacteria and an increase in the number of pathogenic bacteria, have been proven. Gut dysbiosis is closely related to gastrointestinal symptoms and the severity of the disease of COVID-19.
Conclusion: To determine whether these changes in the composition of the intestinal microbiota reflect the cause or effect of COVID-19, it is necessary to perform additional research. Therapeutic intervention on the microbiota through probiotics, prebiotics and fecal transplantation is one of the promising strategies for the treatment of COVID-19 in the future, but further research is necessary
Preparation and characterisation of spray-dried sodium hyaluronate and mannitol microspehers
No full textSušenje raspršivanjem često je korišten tehnološki postupak u razvoju lijekova zbog jednostavnosti i mogućnosti pripravljanja produkata kontroliranih svojstava optimiranjem procesnih parametara postupka. Produkti sušenja raspršivanjem su prašci koji se koriste u razne svrhe, između ostalog i za razvoj nazalnih farmaceutskih oblika. Nazalnom primjenom lijekova zaobilazi se metabolizam prvog prolaska kroz jetru kao i krvno-moždana barijera što omogućuje dostavu lijeka izravno u središnji živčani sustav. Inovativni oblici za nazalnu primjenu koriste in situ gelirajuće polimere s ciljem duljeg zadržavanja lijeka na nosnoj sluznici te kontroliranog oslobađanja.
U ovom radu pripravljene su mikrosfere natrijevog hijaluronata i manitola metodom sušenja raspršivanjem te su određena njihova fizičko – kemijska svojstva. U otopinama za sušenje varirana je koncentracija manitola (5 %, 6 %, 7 %, 8 %, 10 %, m/V), dok je koncentracija natrijevog hijaluronata bila konstantna pri 0,5 % (m/V). Iskorištenje procesa sušenja raspršivanjem bilo je u rasponu od 39,4 % do 45,1 %. Sadržaj vlage bio je i iznosio je od 1,25 % ± 0,50 % do 1,98 % ± 0,52 %. Volumni promjeri mikrosfera, Dv10, Dv50 i Dv90, redom su iznosili 9,7 ± 0,1 – 11,3 ± 0,1 μm, 18,3 ± 0,1 – 21,1 ± 0,2 μm i 31,7 ± 0,8 – 44,5 ± 1,4 μm. Najveći Dv90 zabilježen je za mikrosfere iz otopine s 10 % manitola (m/V), dok je uzorak s 5 % manitola u otopini za sušenje (m/V) imao najveći udio čestica promjera većeg od 10 μm. Svojstva tečenja uzoraka prikazana su Hausnerovim omjerom koji je bio u rasponu od 1,565 ± 0,037 do 1,857 ± 0,045. Negativan zeta potencijal mikrosfera (−23,4 ± 1,7 - −8,6 ± 2,0 mV) očekivan je zbog negativno nabijenih karboksilnih skupina natrijeva hijaluronata. Razvijene mikrosfere apsorbirale su znatno veći volumen pročišćene vode u odnosu na apsorbirani volumen simuliranog nosnog fluida tijekom procesa bubrenja. S porastom udjela manitola u mikrosferama, apsorbiran je veći volumen medija za bubrenje.Spray drying is a commonly used technique in drug development due to its simplicity and the possibility of preparing products with controlled properties by optimizing the process parameters. Spray-dried products are powders that are used for various purposes, among others, for nasal drug delivery systems development. Nasal drug administration bypasses the first pass liver metabolism as well as the blood-brain barrier, which enables the delivery of the drug directly to the central nervous system. Innovative pharmaceutical forms for nasal administration use in situ gelling polymers to achieve longer retention of the drug on the nasal mucosa and its controlled release. In this work, microspheres of sodium hyaluronate and mannitol were prepared by the spray drying method and their physical and chemical properties were determined. In the drying solutions, the concentration of mannitol was varied (5%, 6%, 7%, 8%, 10%, m/v), while the concentration of sodium hyaluronate was constant at 0.5% (m/v). The efficiency of the spray drying process ranged from 39.4% to 45.1%. The moisture content was and amounted to 1.25% ± 0.50% to 1.98% ± 0.52%. The volume diameters of the microspheres, Dv10, Dv50 and Dv90, were respectively 9.7 ± 0.1 – 11.3 ± 0.1 μm, 18.3 ± 0.1 – 21.1 ± 0.2 μm and 31.7 ± 0.8 – 44.5 ± 1.4 μm. The highest Dv90 was recorded for microspheres from a solution with 10% mannitol (m/v), while the sample with 5% mannitol in the drying solution (m/v) had the highest proportion of particles with a diameter greater than 10 μm. The flow properties of the samples were shown by the Hausner ratio, which ranged from 1.565 ± 0.037 to 1.857 ± 0.045. The negative zeta potential of the microspheres (−23.4 ± 1.7 - −8.6 ± 2.0 mV) is expected due to the negatively charged carboxyl groups of sodium hyaluronate. The developed microspheres absorbed a significantly larger volume of purified water compared to the absorbed volume of simulated nasal fluid during the swelling process. With an increase in the mannitol ratio in the microspheres, a larger volume of the swelling medium was absorbed
Impact of CYP2C19 polymorphisms and CYP2C haplotype on the clinical outcomes of clopidogrel therapy
No full textZbog velike pojavnosti kardiovaskularnih bolesti, klopidogrel je često propisivan lijek. Kao i kod svakog lijeka važno je pogoditi optimalnu dozu kako pacijent ne bi bio poddoziran i tako imao povećan rizik od razvoja nuspojava poput moždanog udara i infarkta miokarda. No, isto tako može doći i to predoziranja i time rizika od krvarenja. Budući da je klopidogrel prolijek, kako bi ispoljio svoj učinak mora se aktivirati. Jedan od važnijih enzima koji sudjeluje u aktivaciji je enzim CYP2C19. Ovaj enzim je važan za metabolizam više od 30 lijekova te je njegova polimorfnost povezana s velikim interindividualnim varijacijama u terapijskom odgovoru. Novije studije pokazale su mogući utjecaj novog haplotipa CYP2C:TG na aktivnost enzima CYP2C19 i posljedično brzinu aktivacije klopidogrela. U ovom radu ispitivana je povezanost polimorfizama gena CYP2C19 (rs4244285 i rs12248560) i CYP2C (rs2860840 i rs 11188059) s nuspojavama krvarenja ili neučinkovitosti klopidogrela. Genotipizirano je 145 ispitanika koji su primali klopidogrel kao dio terapije različitih kardiovaskularnih i cerebrovaskularnih bolesti. Genotipizacija je učinjena metodom TaqMan®. Pokazana je statistički značajna povezanost između nefunkcionalnog alela (CYP2C19*2) i neučinkovitosti klopidogrela (p=0,043), što se slaže s mnogobrojnim studijama koje su to dokazale. Naspram tome, nije dokazana povezanost alela povećane funkcije CYP2C19*17 i novog haplotipa CYP2C:TG niti s neučinkovitošću, niti sa pojavom krvarenja. Potrebna su daljnja istraživanja na većem broju ispitanika kako bi se ispitala povezanost polimorfizma ovog enzima i haplotipa CYP2C:TG s varijacijama u terapijskom odgovoru na klopidogrel.Due to the high occurrence of cardiovascular diseases, clopidogrel is a frequently prescribed drug. As with any drug, it is important to find the optimal dose so that the patient is not underdosed and thus has an increased risk of developing side effects such as stroke and myocardial infarction. However, there can also be an overdose and thus the risk of bleeding. Because clopidogrel is a prodrug, it must be activated in order to exert its effect. One of the most important enzymes involved in activation is the CYP2C19 enzyme. This enzyme is important for more than 30 drugs, and its polymorphisms are associated with large interindividual variations in therapeutic response. Recent studies have shown a possible influence of the new haplotype CYP2C:TG on CYP2C19 enzyme activity and consequently clopidogrel activation. This study investigated the association of CYP2C19 (rs4244285 and rs12248560) and CYP2C (rs2860840 and rs 11188059) gene polymorphisms with the side effect of bleeding or ineffectiveness of clopidogrel. 145 subjects who received clopidogrel as part of therapy for various cardiovascular and cerebrovascular diseases were genotyped. Genotyping was done using the TaqMan® method. A statistically significant association between the allele without function (CYP2C19*2) and the ineffectiveness of clopidogrel was shown (p=0.043), which is in-line with several studies that have proven this. In contrast, we were unable to prove the association of the allele of accelerated CYP2C19*17 function and the new haplotype CYP2C:TG neither with ineffectiveness nor with the occurrence of bleeding. Further research is needed on a larger number of subjects in order investigate the association of the polymorphism of this enzyme and the CYP2C:TG haplotype with variability in clopidogrel therapeutic response
The Impact of the COVID-19 Pandemic on the Occurrence of Medication Errors
No full textCilj ovog rada bio je ispitati utjecaj pandemije na učestalost nastanka i karakteristike medikacijskih pogrešaka. Prijave medikacijskih pogrešaka su analizirane prema vrsti prijavitelja, dobi i spolu pacijenata, raspodjeli suspektnih lijekova prema ATK klasifikaciji, raspodjeli nuspojava prema djelatnim tvarima, ozbiljnosti prijava i njihovom kriteriju ozbiljnosti te prijavljenim vrstama medikacijskih pogrešaka. Analizirano je ukupno 12 349 nuspojava prijavljenih prije pandemije (1.1.2017.-31.12.2019.) te 12 391 prijava tijekom pandemije(1.1.2020.-31.12.2022.). Rezultati upućuju na značajno povećanje broja medikacijskih pogrešaka tijekom pandemije, kao i na porast broja nuspojava prijavljenih od strane pacijenata, što se može objasniti smanjenom dostupnošću zdravstvenih radnika i posljedičnom nemogućnosti savjetovanja pacijenata o ispravnom načinu primjene lijekova.The aim of this study was to examine the impact of the pandemic on the frequency and characteristics of medication errors. Reports of medication errors were analyzed according to the type of reporter, the age and gender of patients, the distribution of suspect drugs according to ATC classification, the distribution of side effects according to active substances, the seriousness of reports and their seriousness criteria, and the reported types of medication errors. A total of 12,349 side effects reported before the pandemic (January 1, 2017 - December 31, 2019) and 12,391 reports during the pandemic (January 1, 2020 - December 31, 2022) were analyzed. The results indicate a significant increase in the number of medication errors during the pandemic, as well as an increase in the number of side effects reported by patients, which can be explained by the reduced availability of healthcare workers and the consequent inability to advise patients on the proper use of medications
Synthesis and characterization of tafamidis amide prodrug
No full textOvaj rad dio je istraživanja provedenog u Zavodu za farmaceutsku kemiju Farmaceutsko-biokemijskog fakulteta Sveučilišta u Zagrebu u sklopu projekta Uloga transtiretina u leptomeningealnoj i cerebrovaskularnoj amiloidozi povezanoj sa sporadičnom Alzheimerovom bolesti i neuroprotektivni potencijal prolijeka tafamidisa – TransADamis Medicinskog fakulteta Sveučilišta u Zagrebu koji obuhvaća sintezu i in vivo ispitivanje sposobnosti amidnog prolijeka tafamidisa da stabilizira transtiretin (TTR) u središnjem živčanom sustavu i ublaži streptozotocinom induciranu neurodegeneraciju i kognitivni deficit.
Cilj ovog rada bila je sinteza i karakterizacija amidnog prolijeka tafamidisa. Sinteza je započeta prevođenjem karboksilne kiseline tafamidisa do odgovarajućeg piperazinskog amida 1 s tert-butiloksikarbonilnom zaštitnom skupinom nakon čega je zaštitna skupina hidrolizirana čime je dobiven konačni produkt 2.
Strukture svih sintetiziranih spojeva potvrđene su uobičajenim analitičkim i spektroskopskim metodama (FT-IR, 1H i 13C NMR, MS), a relativna čistoća određena je pomoću tekućinske kromatografije visoke djelotvornosti (HPLC). Konačni produkt u potpunosti zadovoljava Lipinskijeva i Veberova pravila koja se koriste za predviđanje oralne bioraspoloživosti lijekova. Također, web alat SwissADME predviđa visoku oralnu bioraspoloživost i mogućnost prelaska KMB-a amida 2 što je u skladu s rezultatima prethodno objavljene studije in vivo.
Učinak amida 2 na stabilizaciju TTR-a u središnjem živčanom sustavu i ublažavanje neurodegeneracije bit će ispitan u daljnjim istraživanjima koja prelaze okvire ovog rada.This work is part of the research conducted at the Department of Pharmaceutical Chemistry, University of Zagreb Faculty of Pharmacy and Biochemistry, within the project Role of Transthyretin in Leptomeningeal and Cerebrovascular Amyloidosis Associated with Sporadic Alzheimer's Disease and the Neuroprotective Potential of Tafamidis Prodrug – TransADamis of the School of Medicine, University of Zagreb, which encompasses the synthesis and in vivo testing of the ability of the amide prodrug of tafamidis to stabilize transthyretin (TTR) in the central nervous system and mitigate streptozotocin-induced neurodegeneration and cognitive deficit.
The aim of this thesis was the synthesis and characterization of the amide prodrug of tafamidis. The synthesis began with the conversion of tafamidis carboxylic acid to the corresponding piperazine amide 1 with a tert-butoxycarbonyl protecting group, followed by hydrolysis of the protecting group to obtain the final product 2.
The structures of all synthesized compounds were confirmed by standard analytical and spectroscopic methods (FT-IR, 1H and 13C NMR, MS), and the relative purity was determined by high-performance liquid chromatography (HPLC). The final product fully complies with Lipinski's and Veber's rules used to predict the oral bioavailability of drugs. Additionally, the web tool SwissADME predicts high oral bioavailability and the ability of amide 2 to cross the blood-brain barrier, which is consistent with the results of a previously published in vivo study. The effect of amide 2 on the stabilization of TTR in the central nervous system and the mitigation of neurodegeneration will be examined in further research beyond the scope of this paper