University of Zagreb
Repository of Faculty of Pharmacy and Biochemistry University of ZgrebNot a member yet
2946 research outputs found
Sort by
Freeze-drying of melatonin nanosuspension in the presence of a cryoprotectant
No full textMelatonin je hormon lipofilnog karaktera i ograničene topljivosti u vodi koji u ljudskom organizmu fiziološki sintetizira epifiza. Ostvaruje pleiotropni učinak na glavne fiziološke funkcije, stanični ciklus i biokemijske mehanizme s cirkadijanim obilježjima. Terapijska primjena se primarno odnosi na poremećaje spavanja poput nesanice. Suvremena istraživanja usmjerena su na razvoj inovativnih farmaceutskih oblika melatonina. Izrada nanosuspenzija predstavlja jednu od strategija oblikovanja slabo topljivih lijekova s ciljem povećanja topljivosti, brzine otapanja i bioraspoloživosti. Međutim, glavno ograničenje je dugoročna nestabilnost nanosuspenzija i sklonost čestica agregaciji. Sušenje smrzavanjem, odnosno liofilizacija, pokazalo se kao jedna od metoda razvoja čvrstih oblika nanosuspenzija povećane fizičke stabilnosti. Cilj ovog rada bio je ispitati mogućnost liofilizacije nanosuspenzije melatonina u svrhu razvoja suhog oblika koristeći glukozu, manitol i trehalozu (0,5 – 2,5 %, m/V) kao krioprotektore, te ispitati njihov utjecaj na kratkoročnu i dugoročnu stabilnost razvijenih liofilizata mjereći veličinu, indeks polidisperznosti i zeta-potencijal nanokristala u rekonstituiranim suspenzijama. Nanosuspenzija melatonina (0,143 %, m/V) pripravljena je metodom vlažnog mljevenja uz dodatak polisorbata 80 (0,286 %, m/V) kao površinski aktivne tvari, u masenom omjeru 1:2. Veličina čestica ishodne nanosuspenzije iznosila je 176,4 ± 3,3 nm uz indeks polidisperznosti od 0,141 ± 0,013. Liofilizacija nanosuspenzije melatonina uz dodatak manitola rezultirala je liofilizatom volumena jednakog volumenu suspenzije prije liofilizacije. U slučaju trehaloze uočen je djelomični kolaps, a u slučaju glukoze potpuni kolaps strukture liofilizata. Manitol i trehaloza su pri koncentracijama od 0,5 do 2,5 % (m/V) osigurali zadovoljavajuće očuvanje fizičkih svojstava nanokristala melatonina nakon liofilizacije, za razliku od glukoze. Porastom koncentracije manitola i trehaloze u liofiliziranoj nanosuspenziji djelomično se povećala uspješnost očuvanja veličine i raspodjele veličina nanokristala. Tijekom skladištenja pri 4 °C u razdoblju od 60 dana, uz manitol i trehalozu (2,5 %, m/V) kao krioprotektore, uspješno su sačuvani izgled liofilizata i fizička svojstva nanokristala nakon rekonstitucije, čime je potvrđen njihov krioprotektivni učinak u procesu liofilizacije nanosuspenzije melatonina.Melatonin is a lipophilic hormone with limited water solubility, which is physiologically synthesized in the human body by the pineal gland. It has a pleiotropic effect on the main physiological functions, the cell cycle, and biochemical mechanisms with circadian characteristics. Therapeutic use is primarily related to sleep disorders such as insomnia. Nowadays, research is focused on the development of innovative pharmaceutical forms of melatonin. The production of nanosuspensions represents one of the strategies for the formulation of poorly soluble drugs with the aim of increasing their solubility, dissolution rate and bioavailability. However, the main limitation is the long-term instability of nanosuspensions and the tendency of particles to aggregate. Freeze drying, i.e. lyophilisation, proved to be one of the methods of developing solid forms of nanosuspensions with increased physical stability. The aim of this work was to examine the possibility of lyophilisation of melatonin nanosuspension for the purpose of developing a dry form using glucose, mannitol and trehalose (0.5-2.5%, w/v) as cryoprotectants, and to examine their influence on the short-term and long-term stability of the developed lyophilisates by measuring size, polydispersity index and zeta-potential of nanocrystals in reconstituted suspensions. Nanosuspension of melatonin (0.143%, w/v) was prepared by the method of wet milling with the addition of polysorbate 80 (0.286%, w/v) as a surfactant, in a weight ratio of 1:2. The particle size within the initial nanosuspension was 176.4 ± 3.3 nm with a polydispersity index of 0.141 ± 0.013. Lyophilisation of the melatonin nanosuspension in the presence of mannitol resulted in a lyophilisate volume equal to the volume of the suspension before lyophilisation. In the case of trehalose, a partial collapse was observed, and in the case of glucose, a complete collapse of the structure of the lyophilisate was observed. Mannitol and trehalose at concentrations from 0.5 to 2.5 % (w/v) ensured satisfactory preservation of the physical properties of melatonin nanocrystals after lyophilisation, in contrast to glucose. Increasing the concentration of mannitol and trehalose in the lyophilised nanosuspension partially increased the success of preserving the size and size distribution of nanocrystals. During storage at 4 °C for a period of 60 days, with mannitol and trehalose (2.5 %, w/v) as cryoprotectants, the appearance of the lyophilisate and the physical properties of the nanocrystals after reconstitution were successfully preserved, thus confirming their cryoprotective effect in the process of lyophilisation of melatonin nanosuspensions
Immunosuppresive therapy and potentially clinically significant interactions in heart transplant patients
No full textPacijenti s transplantiranim srcem rizična su skupina za nastanak interakcija lijekova. Kompliciranost zahvata i politerapija povećavaju rizik od pojavnosti stvarnih interakcija lijekova. Interakcije lijekova kompliciraju tijek liječenja, povećavaju rizik od hospitalizacije i produljene hospitalizacije te ugrožavaju sigurnost pacijenta. Jedan od ključnih faktora za dugoročno preživljavanje pacijenata nakon transplantacije je odgovarajuća imunosupresivna terapija. Cilj ovog rada bio je utvrditi potencijalno klinički značajne interakcije lijekova u pacijenata s transplantiranim srcem te objasniti mehanizme koji ih uzrokuju. U istraživanje je uključeno 20 pacijenata s transplantiranim srcem zaprimljenih na Zavod za bolesti srca i krvnih žila, Klinička bolnica Dubrava. Za utvrđivanje podataka o pacijentima i farmakoterapiji korišten je bolnički informacijski sustav i Najbolja moguća medikacijska povijest. Politerapija je definirana kao istovremena primjena 5-9 lijekova, dok je primjena 10 ili više lijekova analizirana kao prekomjerna politerapija. Potencijalno klinički značajne interakcije analizirane su pomoću softverskog programa Lexicomp. Interakcije su također podijeljene i prema mehanizmu djelovanja. Najčešće zabilježene dijagnoze u pacijena ta bila su
hipertenzija, ishemične bolesti srca, hiperlipidemija, kronično bubrežno zatajenje, šećerna bolest i kronični gastritis. Najmanje jedna interakcija C stupnja bila je prisutna kod svih pacijenata, dok je 60 % pacijenata imalo barem jednu D interakciju. Najčešća interakcija bila je C stupnja kliničke značajnosti između MMF-a i pantoprazola, zabilježena kod 70 % pacijenata. Ovo istraživanje pokazalo je da su u utvrđenim interakcijama lijekova imunosupresivi bili najčešće zastupljeni. U interakcijama C stupnja najzastupljeniji imunosupresivni lijek bio je takrolimus s udjelom od 10,7%, a u D interakcijama ciklosporin s 16,1 %. Interakcije imunosupresiva češće su bile farmakodina mičke (50 %) dok je farmakokinetičkih bilo 28,4 %. Međusobne interakcije imunosupresiva bile su zastupljene s udjelom od 15,7 %. Interakcije imunosupresiva s ostalim lijekovima najčešće su utvrđene s ATK skupinom A. Lijekovi iz ATK skupine A najčešće zastupljeni u interakcijama bili su oralni antidijabetici, IPP-ovi te inzulini i analozi. Utvrđeni rezultati naglašavaju potrebu za stalnim praćenjem transplantiranih pacijenata. S obzirom na moguće posljedice osobitu pozornost treba obratiti na mogućnost smanjenja koncentracije imunosupresiva, nefrotok sičnost, hiperkalijemiju, hiperglikemiju, gastrointestinalnu toksičnost i hiperuricemiju. Klinički farmaceuti trebaju imati aktivnu ulogu u prepoznavanju
i upravljanju interakcijama lijekova posebice kod svakog novog propisivanja lijeka u pacijenata s transplatiranim srcem.Patients with heart transplants are a high-risk group for drug interactions. The complexity of the procedure and polytherapy increase the risk of actual drug interactions. Drug interactions complicate the course of treatment, increase the risk of hospitalization and prolonged hospital stays, and endanger patient safety. One of the key factors for long-term survival of patients after transplantation is appropriate immunosuppressive therapy. The aim of this study was to identify potentially clinically significant drug interactions in heart transplant patients and to explain the mechanisms that cause them. The study included 20 heart transplant patients admitted to the Department of Cardiology and Vascular Diseases, University Hospital Dubrava. Data on patients and pharmacotherapy were obtained using the hospital information system and the Best Possible Medication History. Polytherapy was defined as the simultaneous use of 5-9 drugs, while the use of 10 or more drugs
was analyzed as excessive polytherapy. Potentially clinically significant interactions were analyzed using the Lexicomp software program. Interactions were also classified according to their mechanism of action. The most frequently recorded diagnoses in patients were hypertension, ischemic heart disease, hyperlipidemia, chronic renal failure, diabetes mellitus, and chronic gastritis. At least one grade C interaction was present in all patients, while 60% of patients had at least one grade D interaction. The most common interaction was a grade C clinically significant interaction between MMF and pantoprazole, recorded in 70% of patients. This study showed that immunosuppressants were most commonly involved in the identified drug interactions. In grade C interactions, the most common immunosuppressive drug was tacrolimus,
accounting for 10.7%, while in grade D interactions, it was cyclosporine, accounting for 16.1%. Immunosuppressant interactions were more frequently pharmacodynamic (50%), while pharmacokinetic interactions accounted for 28.4%. Mutual interactions among immunosuppressants accounted for 15.7% of cases. Interactions between immunosuppressants and other drugs were most commonly identified with the ATC group A. The most commonly involved drugs from ATC group A in interactions were oral antidiabetics, PPIs, and insulin and its analogs. The findings underscore the need for continuous monitoring of transplant patients. Given the potential consequences, special attention should be paid to the possibility of reduced immunosuppressant concentrations, nephrotoxicity, hyperkalemia, hyperglycemia, gastrointestinal toxicity, and hyperuricemia. Clinical pharmacists should play an active role in identifying and managing drug interactions, especially when a new drug is prescribed to heart transplant patients
The effect of storage temperature on the phenolic composition of olive leaves and branches
No full textNakon rezidbe stabala masline nastaje mnogo otpada u obliku lišća i grančica koji su bogati
polifenolima. U sklopu ovog diplomskog rada izrađeni su suhi ekstrakti lista masline, grančice s
listom te grančice bez lista. Biljni materijal sušen je na zraku i zatim čuvan na sobnoj temperaturi i
na 4°C tri mjeseca prije ekstrakcije. Nakon ekstrakcije pomoću ultrazvučne kupelji i pripreme suhih
ekstrakata, analiziran je sadržaj ukupnih polifenola (TPC), o-difenola (o-DPC) i ukupnih flavonoida
(TFC) UV-Vis metodom te oleuropeina (OLE) HPLC metodom. Uspoređen je sadržaj navedenih
tvari u osušenim uzorcima čuvanim na sobnoj temperaturi i onima na 4°C. Ekstrakt lista pripremljen
iz lišća skladištenog na 4°C imao je najveći sadržaj OLE, 30,7 mg/g suhog ekstrakta (DE), a i
rezultati izraženi po masi suhog biljnog uzorka (DW) pokazali su najveći sadržaj OLE u listu
skladištenom na 4°C, 2,88 mg/g DW. Sadržaj TPC i TFC bio je najveći u ekstraktu grančica bez
lišća skladištenima na 4°C s vrijednostima od 193 mg GAE/g DE, odnosno 82,9 mg CE/g DE).
Rezultati izraženi po DW, pokazali su najveći sadržaj u listu skladištenom na 4°C s vrijednostima
TPC od 14,1 mg GAE/g DW te TFC od 5,99 mg CE/g DW. Sadržaj o-DPC bio je najveći u ekstraktu
lista na 4°C, 150 mg GAE/g DE, te u listu na 4°C, 14,1 mg GAE/g DE. Ovaj je rad pokazao da
temperatura skladištenja biljnoga materijala ima utjecaj na vrijednosti TPC, o-DPC, TFC i posebice
OLE, čiji je sadržaj bio skoro dvostruko niži u uzorcima skladištenima na sobnoj temperaturi.After pruning olive trees, a significant amount of waste is generated in the form of leaves and
branches, which are rich in polyphenols. As part of this thesis, dry extracts were prepared from olive
leaves, branches with leaves, and branches without leaves. The plant material was air-dried and then
stored at room temperature and at 4°C for three months before extraction. After extraction using an
ultrasonic bath and preparation of dry extracts, the total polyphenol content (TPC), o-diphenol
content (o-DPC), and total flavonoid content (TFC) were analyzed using the UV-Vis method, while
oleuropein (OLE) was analyzed using the HPLC method. The content of these compounds in the
dried samples stored at room temperature and ones at 4°C was then compared. The leaf extract
prepared from leaves stored at 4°C had the highest OLE content, 30.7 mg/g of dry extract (DE), and
the results expressed per dry plant material (DW) showed the highest OLE content in leaves stored
at 4°C, 2.88 mg/g DW. The TPC and TFC content was highest in the extract from branches without
leaves stored at 4°C, with values of 193 mg GAE/g DE and 82.9 mg CE/g DE, respectively. Results
expressed per DW showed the highest content in leaves stored at 4°C, with TPC values of 14.1 mg
GAE/g DW and TFC of 5.99 mg CE/g DW. The o-DPC content was highest in the leaf extract stored
at 4°C, 150 mg GAE/g DE, and in leaves at 4°C, 14.1 mg GAE/g DW. This study showed that the
storage temperature of plant material affects the values of TPC, o-DPC, TFC, and especially OLE,
whose content was nearly half as low in samples stored at room temperature
Synthesis of O-benzyl oxime quinuclidines and their anticholinesterase activity
No full textU okviru ovog rada klasičnim postupcima organske sinteze pripravljen je O-benzilni eter kinuklidin-
3-on oksima reakcijom hidroklorida kinuklidin-3-ona i O-benzilhidroksilamina, bez dodatka
anorganskih ili organskih baza. Kvaternizacijom kinuklidinskog atoma dušika u molekuli
pripravljenog O-benziloksima s nesupstituiranim i odabranim, različito supstituiranim benzilbromidima
(trifluormetilna ili trifluormetoksi skupina u para položaju, tert-butilna skupina u para
ili meta položajima, spojevi halogena na određenim pozicijama prstena) pripravljen je niz od devet
spojeva, od kojih osam do sada nisu opisani u literaturi. Novosintetiziranim spojevima određeno je
talište te je struktura određena i potvrđena spektroskopskim metodama: infracrvenom
spektroskopijom, 1D nuklearnom magnetskom rezonancijom te spektrometrijom masa. Nakon
sinteze te fizikalno-kemijske karakterizacije spojeva, svima je određen inhibitorni potencijal prema
enzimu butirilkolinesterazi iz konjskog seruma (EC 3.1.1.8) korištenjem Ellmanove kolorimetrijske
metode za određivanje aktivnosti acetilkolinesteraze. Svi ispitani spojevi reverzibilno su inhibirali
enzim u mikro- i nanomolarnom području. Kao najsnažniji inhibitor pokazao se spoj 6, kvaterni
derivat oksima s dvije tert-butilne skupine u meta položajima na aromatskom prstenu benzilne
skupine vezane na kvaterni dušikov atom kinuklidina (IC50 = 0,28 ± 0,01 μmol dm-3).Within this work, O-benzyl ether quinuclidin-3-one oxime was prepared by the reaction of
hydrochloride salts of quinuclidin-3-one and O-benzylhydroxylamine, without the addition of
inorganic or organic bases. By quaternization of a quinuclidine nitrogen atom in a molecule prepared
with O-benzyl oxime with unsubstituted and selected, differently substituted benzyl bromides
(trifluoromethyl or trifluoromethoxy groups in the para- position, tert-butyl group in the para or meta
positions, halogen compounds at certain benzene ring positions), a series of nine compounds,
quaternary derivatives of quinuclidine, was prepared. Out of those nine, eight have not been
described in the literature so far. The melting point of the newly synthesized compounds was
determined, and the structure was characterized and confirmed by spectroscopic methods: infrared
spectroscopy, 1D nuclear magnetic resonance and mass spectrometry. Subsequent to physicochemical
characterization of the compounds, the inhibitory potential toward the enzyme
butyrylcholinesterase from equine serum (EC 3.1.1.8) was determined using the Ellman colorimetric
method to determine cholinesterase activity. All tested compounds reversibly inhibited the enzyme
in the micromolar and nanomolar range. The most potent inhibitor was quaternary O-benzyl oxime
derivative with two tert-butyl groups in meta positions on the quaternary benzyl group (IC50 = 0.28
± 0.01 μmol dm-3)
Application of cyclodextrin in the development of topical preparations for the skin
No full textCiklodekstrini su prirodni ciklički oligosaharidi koji su našli široku primjenu u dermatološkim i kozmetičkim formulacijama zbog svoje jedinstvene sposobnosti stvaranja inkluzijskih kompleksa. Ova svojstva omogućuju poboljšanje stabilnosti, topljivosti i kontroliranog otpuštanja aktivnih sastojaka, čime se povećava učinkovitost i sigurnost pripravaka za kožu. U kozmetičkim proizvodima, ciklodekstrini se koriste za optimizaciju performansi krema protiv starenja, zaštitnih krema za sunčanje i hidratantnih preparata, čime se povećava učinkovitost i produljuje djelovanje aktivnih tvari na koži. U dermatološkim aplikacijama, ciklodekstrini su ključni u formulaciji hidrogelova za zacjeljivanje rana. Ovi hidrogelovi omogućuju održavanje optimalne vlažnosti i kontrolirano otpuštanje terapeutskih agenasa, što ubrzava proces regeneracije tkiva i smanjuje rizik od infekcija. Hidrogelovi bazirani na hidroksipropil-β-ciklodekstrinu pokazali su izvrsne rezultate u kliničkim ispitivanjima, značajno poboljšavajući brzinu zacjeljivanja rana.
Regulativni okviri za upotrebu ciklodekstrina variraju širom svijeta. U Sjedinjenim Američkim Državama, kozmetički proizvodi koji sadrže ciklodekstrine ne zahtijevaju odobrenje FDA, osim ako sadrže specifične boje, dok Europska unija i Kanada imaju strože smjernice koje definiraju sigurne koncentracije ciklodekstrina u kozmetičkim i dermatološkim proizvodima.
Unatoč njihovim prednostima, ciklodekstrini mogu uzrokovati iritaciju kože pri visokim koncentracijama, što naglašava potrebu za daljnjim istraživanjima kako bi se osigurala njihova sigurnost. Daljnji razvoj formulacija i usklađivanje s regulativnim zahtjevima ključni su za širu primjenu ciklodekstrina u industriji njege kože.Cyclodextrins are natural cyclic oligosaccharides widely applied in dermatological and cosmetic formulations due to their unique ability to form inclusion complexes. These properties make it possible to improve the stability, solubility and controlled release of active ingredients, thereby increasing the effectiveness and safety of skin preparations. In cosmetic products, cyclodextrins are used to optimize the performance of anti-ageing creams, sunscreens and moisturizers, thereby increasing the efficacy and prolonging the action of active substances on the skin. In dermatological applications, cyclodextrins are crucial in formulating hydrogels for wound healing. These hydrogels enable the maintenance of optimal humidity and the controlled release of therapeutic agents, which accelerates the process of tissue regeneration and reduces the risk of infections. Hydrogels based on hydroxypropyl-β-cyclodextrin) have shown excellent results in clinical trials, significantly improving the speed of wound healing.
Regulatory frameworks for the use of cyclodextrins vary around the world. In the United States, cosmetic products containing cyclodextrins do not require FDA approval unless they contain specific colours. At the same time, the European Union and Canada have stricter guidelines that define safe concentrations of cyclodextrins in cosmetic and dermatological products.
Despite their benefits, cyclodextrins can cause skin irritation at high concentrations, highlighting the need for further research to ensure their safety. Further formulation development and compliance with regulatory requirements are critical to the broader use of cyclodextrins in the skin care industry
Dietary supplements and supportive therapy in treatment of depression
No full textOvaj diplomski rad bavi se unapređenjem liječenja depresije, jedne od najčešćih mentalnih bolesti koja pogađa oko 5% svjetske populacije. Integracija dodataka prehrani i suportivnih terapija u standardnu farmakoterapiju može biti od ključne važnosti u ovom kontekstu. Iako su antidepresivi osnovni stup terapije, njihov spor početak djelovanja, nuspojave i dugotrajnost često rezultiraju smanjenom adherencijom pacijenata. Kako bi se odgovorilo na ove izazove, rad istražuje učinkovitost i sigurnost različitih dodataka prehrani poput L-5-hidroksitriptofana, eikozapentaenske kiseline, folne kiseline, šafrana i kurkumina, koji pokazuju potencijal u ublažavanju simptoma depresije i smanjenju nuspojava. Također se analiziraju suportivne terapije kao što su akupunktura, art terapija, terapija glazbom, svjetlosna terapija, mindfulness, relaksacijska terapija i joga, koje doprinose poboljšanju općeg stanja pacijenata i njihove kvalitete života.
Integracija ovih terapija u postojeće liječenje može značajno povećati učinkovitost terapije, smanjiti rizik od prekida liječenja i osigurati dugotrajniju remisiju. Stoga se predlaže multidisciplinarni pristup koji objedinjuje farmakološke i nefarmakološke metode, pružajući pacijentima individualiziran plan liječenja. Ovakav pristup je od iznimne važnosti za sveobuhvatno ljekarničko savjetovanje.This thesis explores the enhancement of depression treatment, one of the most common mental disorders, affecting around 5% of the global population. The integration of dietary supplements and supportive therapies into standard pharmacotherapy can play a crucial role in this context. Although antidepressants are the cornerstone of therapy, their slow onset of action, side effects, and prolonged duration often lead to reduced patient adherence. To address these challenges, the thesis examines the efficacy and safety of various dietary supplements such as L-5-hydroxytryptophan, eicosapentaenoic acid, folic acid, saffron, and curcumin, which show potential in alleviating depression symptoms and reducing side effects. Additionally, supportive therapies such as acupuncture, art therapy, music therapy, light therapy, mindfulness, relaxation therapy, and yoga are analyzed, as they contribute to improving the overall well-being and quality of life of patients.
The integration of these therapies into existing treatment can significantly enhance therapeutic effectiveness, reduce the risk of treatment discontinuation, and ensure longer-lasting remission. Therefore, a multidisciplinary approach is recommended, combining pharmacological and non-pharmacological methods to provide patients with an individualized treatment plan. This approach is of utmost importance for comprehensive pharmaceutical counseling
Liquid biopsy in lymphoma
No full textLimfomi su maligni tumori limfocitne loze koji se prezentiraju kao tumorske mase prvenstveno u limfoidnim organima. Prema SZO klasifikaciji dijele se u dvije glavne skupine, a u svakoj skupini nalazi se niz obitelji, entiteta/tipova i podtipova s heterogenim kliničkim ishodima i kompleksnim molekularnim karakteristikama koje se mijenjaju tijekom vremena. Konvencionalne metode za dijagnozu i praćenje limfoma su invazivne i tehnički zahtjevne, a heterogenost limfoma dodatno otežava dijagnozu i odabir odgovarajuće terapije. Razvoj tekuće biopsije otvorio je nove mogućnosti za neinvazivnu dijagnostiku, prognozu i praćenje ovih složenih hematoloških maligniteta. To je neinvazivna metoda kojom se iz uzorka krvi ili neke druge tjelesne tekućine izoliraju cirkulirajuće tumorske stanice (CTC), slobodna cirkulirajuća DNA (cfDNA), egzosomi, i cirkulirajuće RNA (miRNA, lncRNA). Tekuća biopsija pruža sveobuhvatnu sliku o heterogenosti limfoma i omogućuje praćenje dinamike tumora u stvarnom vremenu. Ovaj pregledni rad istražuje trenutno stanje i budući potencijal tekuće biopsije u limfomima te ističe njezinu korist u genotipiziranju limfoma, ranom otkrivanju relapsa, procjeni odgovora na liječenje te praćenju minimalne rezidualne bolesti i klonalne evolucije. Napredne metode temeljene na PCR-u i NGS-u značajno su poboljšale osjetljivost i specifičnost testova tekuće biopsije. Cirkulirajuća tumorska DNA (ctDNA) pokazala se kao najperspektivniji i najšire primjenjivani biljeg tekuće biopsije u različitim podtipovima limfoma. MiRNA, egzosomi i lncRNA imaju sposobnost pružanja dodatnih informacija o biologiji limfoma i progresiji bolesti. CTC imaju ograničenu korisnost u većini limfoma zbog njihovog malog broja, ali potencijalno mogu pružiti informacije u leukemijskim fazama bolesti. Unatoč obećavajućim rezultatima, izazovi ostaju u standardizaciji, kliničkoj validaciji i integraciji s postojećim dijagnostičkim i prognostičkim alatima. Stoga je potrebno riješiti navedene izazove prije integracije tekuće biopsije u rutinsku kliničku praksu.Lymphomas are malignant neoplasms of lymphocyte lineage that present as tumor masses primarily involving the lymphoid organs. According to the WHO classification, they are divided into two main groups, with each group containing a range of families, entities/types and subtypes with heterogeneous clinical outcomes and complex molecular characteristics that change over time. Conventional methods for diagnosing and monitoring lymphomas are invasive and technically demanding, and the heterogeneity of lymphomas further complicates diagnosis and selection of appropriate therapy. The development of liquid biopsy has opened up new possibilities for non-invasive diagnosis, prognosis and monitoring of these complex hematological malignancies. It is a non-invasive method that isolates circulating tumor cells (CTCs), cell-free DNA (cfDNA), exosomes and circulating RNAs (miRNA, lncRNA) from a blood sample or other body fluid. Liquid biopsy provides a comprehensive picture of lymphoma heterogeneity and allows real-time monitoring of tumor dynamics. This review paper explores the current state and future potential of liquid biopsy in lymphomas and highlights its benefits in lymphoma genotyping, early detection of relapse, assessment of treatment response, and monitoring of minimal residual disease and clonal evolution. Advanced PCR-based and NGS-based methods have significantly improved the sensitivity and specificity of liquid biopsy tests. Circulating tumour DNA (ctDNA) has emerged as the most promising and widely applicable liquid biopsy marker in various lymphoma subtypes. MiRNA, exosomes, and lncRNA have the ability to provide additional information about lymphoma biology and disease progression. CTCs have limited utility in most lymphomas due to their small numbers, but can potentially provide information in leukemic phases of the disease. Despite promising results, challenges remain in standardization, clinical validation, and integration with existing diagnostic and prognostic tools. Therefore, these challenges need to be addressed before integrating liquid biopsy into routine clinical practice
Advancements in electron paramagnetic resonance (EPR) spectroscopy: A comprehensive tool for pharmaceutical research
No full textElectron paramagnetic resonance (EPR) spectroscopy has long been established across various scientific disciplines for characterizing organic radicals, organometallic complexes, protein structures and dynamics, polymerization processes, and radical degradation phenomena. Despite its extensive utility in these areas, EPR spectroscopy’s application within pharmaceutical science has historically been constrained, primarily due to factors such as high equipment costs, a steep learning curve, complex spectral deconvolution and analysis, and a traditional lack of emphasis on single-electron chemistry in pharmaceutical research. This review aims to provide a thorough examination of EPR spectroscopy’s applications in analyzing a wide array of para-magnetic species relevant to pharmaceutical research. We detail how EPR spectroscopy can be employed to assess free radical scavenging properties in pharmaceutical compounds, elucidate drug mechanisms of action, and explore pharmacokinetics. Additionally, we investigate the role of free radicals in drug-induced toxicity and drug-membrane interactions, while also covering the application of EPR spectroscopy in drug delivery research, advanced studies of metallodrugs, and monitoring of oxygen levels in biological systems through EPR oximetry. The recent advancements in the miniaturization of EPR spectro meters have paved the way for their application in
and
monitoring during the manufacturing process and quality control of pharmaceutical substances and final drug formulations due to being the only direct and non-invasive detection technique for radical detection.
Through these discussions, we highlight the substantial contributions of EPR spectroscopy to the advancement of pharmaceutical sciences
Regulatorni aspekti radiofarmaceutika
No full textCilj istraživanja
Cilj ovog specijalističkog rada je opisati svojstva i primjenu radiofarmaceutika te pridonijeti razumijevanju karakterističnih regulatornih i znanstvenih smjernica za pripremu dokumentacije za davanje odobrenja koje propisuje Europska agencija za lijekove (EMA) i Europska farmakopeja (Ph. Eur.) vezano za razvoj, proizvodnju i provjeru kakvoće radiofarmaceutika.
Materijali i metode
Literatura je pretraživana kroz bibliografske baze podataka prema temi istraživanja, predmetu istraživanja, autorima i časopisu. Analizirani su članci (opći ili specijalizirani) relevantni za problematiku ovoga specijalističkog rada. Pri pretraživanju općih i specijaliziranih članaka korištene su on-line baze podataka (PubMed, ScienceDirect, Elsevier). Regulatorne i znanstvene smjernice detaljno su istražene pretraživanjem internetske baze Europske agencije za lijekove.
U ovom specijalističkom radu obrađene su znanstvene i regulatorne smjernice relevantne za radiofarmaceutike koje se koriste u pripremi dokumentacije za davanje odobrenja za stavljanje lijeka u promet.
Rezultati
S obzirom da se radiofarmaceutici kontroliraju kao lijekovi i kao radioaktivne tvari dva zakonodavna okvira, provjera kakvoće lijeka (dobra proizvođačka praksa, DPP) te sigurnosni propisi (zaštita od zračenja) mogu potencijalno dovesti do konfliktnih situacija. Također smjernice za dobru proizvođačku praksu su razvijene za velike i centralizirane proizvodnje lijekova. Međutim, radiofarmaceutici su često fundamentalno bitno različiti pa te smjernice nisu uvijek u potpunosti primjenjive za njihovu proizvodnju.
Zahtjevi unutar Direktive 2001/83/EZ odnose se i na radionuklidne prekursore tj. radionuklide proizvedene za radioobilježavanje drugih tvari prije primjene. Dakle, principi i zahtjevi dobre proizvođačke prakse odnose se i na proizvođače koji opskrbljuju zdravstvene ustanove ili znanstveno-istraživačke institute radionuklidima namijenjenim za pripravu radiofarmaceutika. U ovom slučaju regulatorni zahtjevi mogu negativno utjecati na dostupnost novih radiofarmaceutika pojavom dodatnih troškova i kašnjenja u opskrbi pa čak i nestašica radionuklida. Stoga se predlaže ažuriranje Direktive 2001/83/EZ kako bi se osigurala veća dostupnost relevantnih radionuklida i smanjili nepotrebni troškovi i kašnjenja s isporukama. Nadalje, Povjerenstvo za humane lijekove (engl. Committee for Medicinal Products for Human Use, CHMP) pri Europskoj agenciji za lijekove pokrenulo je reviziju Smjernice za radiofarmaceutike (oznaka EMEA/CHMP/QWP/306970/2007) koja se smatra jednom od najvažnijih smjernica za kakvoću radiofarmaceutika. S obzirom na to da je prošlo više od 15 godina od posljednje revizije Smjernice, te uzimajući u obzir velik napredak i iskustvo stečeno u proizvodnji i primjeni radiofarmaceutika, kao i činjenicu da su tekstovi Europske farmakopeje u međuvremenu doživjeli značajne promjene, predložena revizija Smjernice o radiofarmaceuticima itekako je potrebna.
Mnoge profesionalne organizacije kao što je Europsko udruženje za nuklearnu medicinu (engl. European Association of Nuclear Medicine, EANM) (neprofitna organizacija) su pokrenule nove projekte kojima će se prikupiti dosadašnje znanje o proizvodnji novih radionuklida i omogućiti razvoj novih regulatornih smjernica s ciljem opskrbe tržišta novim radiofarmaceuticima.
Zaključak
Radiofarmaceutici su radioaktivni lijekovi sa specifičnim karakteristikama za koje su razvijene posebne regulatorne smjernice. Radiofarmaceutski pripravci relativno su novi na tržištu lijekova, a potreba za ovom vrstom lijekova neprestano raste. U proizvodnji radiofarmaceutika
razlikujemo industrijsku proizvodnju velikih razmjera koja se provodi u skladu s dobrom proizvođačkom praksom (DPP) te proizvodnju malih razmjera u zdravstvenim i istraživačkim ustanovama koja se odvija u skladu s dobrom radiofarmaceutskom praksom (DRP). S obzirom na to da se prema Direktivi 2001/83/EZ radiofarmaceutici smatraju lijekovima za primjenu kod ljudi, za njih je potrebno ishoditi odobrenje za stavljanje u promet od nadležnih regulatornih tijela kroz odgovarajuće regulatorne postupke. Za radiofarmaceutike, kao i za ostale humane lijekove, primjenjuju se osnovni postupci odobravanja za stavljanje lijeka u promet (NP, CP, DCP, MRP). Međutim, zbog njihove radioaktivnosti i drugih specifičnih karakteristika, opće regulatorne i znanstvene smjernice koje se primjenjuju za lijekove nisu u potpunosti relevantne za radiofarmaceutike. Zbog toga je važno u razvoju i proizvodnji slijediti specifične smjernice za radiofarmaceutike propisane od strane Europske agencije za lijekove te smjernice regulatornih tijela za nuklearnu medicinu (EANM). Europsko ravnateljstvo za kakvoću lijekova i zdravstvenu skrb (EDQM) objavilo je u Europskoj farmakopeji veći broj monografija koje se specifično odnose na radiofarmaceutske pripravke i polazne materijale, a koje su napisane u svrhu zadovoljavanja potreba regulatornih tijela, proizvođača radiofarmaceutika te svih sudionika uključenih u provjeru kakvoće radiofarmaceutika i njihovih sastavnica. S obzirom na kontinuirani razvoj novih dijagnostičkih i terapijskih radiofarmaceutika, smjernice se i dalje poboljšavaju i mijenjaju. Potrebna je intenzivna suradnja između proizvođača radionuklida, znanstvenika koji razvijaju nove radiofarmaceutike i nadležnih regulatornih tijela kako bi se osigurala dostupnost novih radionuklida i njihova klinička primjenjivost.Objectives
The aim of this professional thesis is to describe peculiarities of radiopharmaceuticals and contribute to the understanding of regulatory and scientific guidelines related to development, production and quality control of radiopharmaceuticals.
Materials and Methods
The literature was reviewed based on the research topic, subject matter, authors, and journals, ranging from general to specialized articles. Selected articles and guidelines pertinent to the focus of the specialist work were included. Online databases such as PubMed, ScienceDirect, Elsevier and the EMA website were utilized to gather the necessary data for this paper.
Results
Given that radiopharmaceuticals are regulated as both medicines and radioactive substances under two legislative frameworks, the quality control of the drug product (Good Manufacturing Practice, GMP) and safety regulations (radiation protection) can potentially lead to conflicting situations. Additionally, the guidelines for Good Manufacturing Practice were developed for large and centralized drug production. However, radiopharmaceuticals are often fundamentally different, so these guidelines are not always fully applicable to their production.
The requirements within Directive 2001/83/EC also apply to radionuclide precursors, i.e., radionuclides produced for the radiolabeling of other substances before administration. Therefore, the principles and requirements of Good Manufacturing Practice also apply to manufacturers who supply radionuclides intended for the preparation of radiopharmaceuticals to healthcare institutions or research institutes. In this case, regulatory requirements can negatively impact the availability of new radiopharmaceuticals by introducing additional costs, delays in supply, and even radionuclide shortages. Thus, an update to Directive 2001/83/EC is
proposed to ensure greater availability of relevant radionuclides and to reduce unnecessary costs and delivery delays. Furthermore, the Committee for Medicinal Products for Human Use (CHMP) at the European Medicines Agency has initiated a revision of the Radiopharmaceuticals Guideline (EMEA/CHMP/QWP/306970/2007), which is considered one of the most important guidelines for the quality of radiopharmaceuticals. Given that more than 15 years have passed since the last revision of the Guideline, and considering the significant progress and experience gained in the production and application of radiopharmaceuticals, as well as the fact that the texts of the European Pharmacopoeia have undergone significant changes in the meantime, the proposed revision of the Radiopharmaceuticals Guideline is indeed necessary. Many professional organizations, such as the European Association of Nuclear Medicine (EANM) (a non-profit organization), have launched new projects to gather existing knowledge on the production of new radionuclides and to enable the development of new regulatory guidelines aimed at supplying the market with new radiopharmaceuticals.
Conclusion
Radiopharmaceuticals are radioactive medicines with specific characteristics for which special regulatory guidelines have been developed. Radiopharmaceutical preparations are relatively new to the drug market, and the demand for this type of medication is continuously increasing. In radiopharmaceutical production, it is distinguished between large-scale industrial production, conducted in accordance with Good Manufacturing Practice (GMP), and small-scale production in healthcare and research institutions, conducted in accordance with Good Radiopharmaceutical Practice (GRP). Since, under Directive 2001/83/EC, radiopharmaceuticals are considered medicinal products for human use, it is necessary to obtain marketing authorization from the relevant regulatory authorities through appropriate regulatory procedures. For radiopharmaceuticals, as well as for other human medicines, the basic marketing authorization procedures (NP, CP, DCP, MRP) apply. However, due to their radioactivity and other specific characteristics, general regulatory and scientific guidelines that
apply to medicines are not fully relevant for radiopharmaceuticals. Therefore, it is essential to follow the specific guidelines for radiopharmaceuticals set by the European Medicines Agency and the guidelines of nuclear medicine regulators (EANM) in their development and production. The European Directorate for the Quality of Medicines and Healthcare (EDQM) has published several monographs in the European Pharmacopoeia that specifically address radiopharmaceutical preparations and starting materials, which have been written to meet the needs of regulatory authorities, radiopharmaceutical manufacturers, and all stakeholders involved in the quality assurance of radiopharmaceuticals and their components. Given the continuous development of new diagnostic and therapeutic radiopharmaceuticals, the guidelines are being constantly improved and updated. Intense collaboration between radionuclide producers, scientists developing new radiopharmaceuticals, and regulatory authorities is needed to ensure the availability of new radionuclides and their clinical applicability
Optimisation of a capillary electrophoretic method for letrozole analysis
No full textZbog velike učestalosti raka dojke i razvoja različitih terapijskih mogućnosti liječenja ove bolesti, od iznimne je važnosti razvoj novih analitičkih metoda za terapijsko praćenje liječenja odabranim lijekovima.
Cilj je ovog istraživanja bio razviti brzu i jednostavnu kapilarnoelektroforetsku metodu za analizu letrozola koja je ekološki prihvatljivija i jeftinija od postojeće HPLC metode. Kako je letrozol u radnim uvjetima analize nenabijen, kao prikladna tehnika odabrana je micelarna elektrokinetička kromatografija.
Analiza je provedena na kapilari od izvučenog kvarca duljine 35 cm, duljine puta do detektora od 27 cm i unutrašnjeg promjera 50 μm. Uzorci su hidrodinamički injektirani u kapilaru pod tlakom od 50 mbar, u vremenu od 4 sekunde. Za detekciju letrozola korištena je valna duljina od 200 i 219 nm.
Tijekom optimizacije metode ispitan je utjecaj koncentracije boratnog pufera (15 – 25 mM) i SDS-a (20 – 65 mM) te temperature (20 – 30 °C) na vrijeme analize i površinu pika. Kao optimalne vrijednosti pokazale su se 15 mM boratni pufer pH 9,3 uz dodatak 35 mM SDS-a.
U daljnjim istraživanjima bit će dodan unutarnji standard kako bi se izbjegli vanjski utjecaji na rezultat analize.Due to high incidence of breast cancer in our population and the developement of new treatment options for this desease it is extremly important to work on deveoping new analytical methods for therapeutic drug monitoring .
The aim of this study was to develop a fast and simple capillary electrophoretic method for the analysis of letrozole, which will also be more environmentally friendly and cheaper than the already known HPLC method. In working environment letrozole is neutral so the best option for his analysis was micellar electrocinetic cromathography.
The analysis was performed on a 35 cm long capillary of fused silica with effective lenght of 27 cm and an innner diameter of 50μm. For the detection of letrozole we used a wavelenghts of 200 and 219 nm.
During the optimisation of method we observed the effect of the concentration of borate buffer (15-25 mM) and SDS (20-65 mM), as well as temperature (20-30 °C) on the migration time and peak area. Considering the experimental results, 15 mM borate buffer, 35 mM SDS and a temperature of 20 °C were taken as optimal values. In further developement of method, it is required to use internal standard in order to eliminate any external effect on the analysis