Razvoj i odobravanje lijekova za dermalnu primjenu

Cilj istraživanja Lijekovi za dermalnu primjenu razlikuju se od lijekova koji su namijenjeni drugim putovima primjene po tome što odabir pomoćnih tvari ovisi o tome je li cilj postići sistemski ili lokalni učinak. U lijekovima za dermalnu primjenu bioraspoloživost djelatne tvari ovisi o njenim fizičko-kemijskim svojstvima, o farmaceutsko-tehnološkim značajkama pripravka, proizvodnom procesu i načinu na koji pacijent primjenjuje lijek. Cilj ovog specijalističkog rada je opisati biofarmaceutske posebnosti lijekova za dermalnu primjenu i pridonijeti razumijevanju smjernica za pripremu dokumentacije za davanje odobrenja koje propisuje Europska agencija za lijekove (EMA), vezano za razvoj lijeka, proizvodnju, provjeru kakvoće i ispitivanja stabilnosti. Materijali i metode Prilikom izrade specijalističkog rada literatura je pretraživana prema temi istraživanja, predmetu istraživanja, autorima i časopisu. Literatura je pretraživana od općih prema specijaliziranim člancima pri čemu su odabrani članci i smjernice relevantne za problematiku rada. Pri pretraživanju literature i prikupljanju relevantnih podataka korištene su on-line baze podataka (PubMed, ScienceDirect, baza lijekova Agencije za lijekove i medicinske proizvode). Rezultati U posljednjih 60 godina razvoj lijekova za dermalnu primjenu sve više napreduje i puno više se zna o djelovanju lijekova na kožu. U današnje vrijeme razvijaju se različiti nosači djelatnih tvari koji su prilagođeni potrebama pacijenata. Također, produbljeno je razumijevanje utjecaja farmaceutskog oblika lijeka na obim isporuke lijeka do ciljanog mjesta djelovanja, te na djelotvornost i sigurnost primjene lijeka. Na tržištu su dostupni brojni sustavi za dermalnu primjenu, ali ograničeni su na određene djelatne tvari. Daljnje napredovanje u razvoju lijekova za dermalnu primjenu ovisi o mogućnosti prevladavanja problema povezanih s permeabilnošću kože i s iritacijom kože uzrokovane djelatnom i/ili pomoćnim tvarima. Razvoj novih tehnologija koje bi povećale permeabilnost i smanjile iritaciju kože, omogućio bi korištenje hidrofilnih molekula, makromolekula i konvencionalnih djelatnih tvari za nove terapijske indikacije u pripravcima za dermalnu primjenu. Zaključak Lijekovi za dermalnu primjenu velik su izazov za proizvođače lijekova s obzirom na kožnu barijeru čija je primarna uloga zaštititi tijelo od vanjskih utjecaja. Stoga je poznavanje strukture i funkcije kože ključno u razvoju lijekova za dermalnu primjenu. Razvoj lijekova za dermalnu primjenu temelji se na prilagodbi prijenosa kroz kožu izborom odgovarajućih podloga i korištenjem suvremenih farmaceutskih oblika za dermalnu primjenu (nanosustavi). Zahtjev za davanje odobrenja za lijekove za dermalnu primjenu u Europskoj uniji može biti podnesen prema Article 8(3) Direktive 2001/83/EC (cjelovita dokumentacija) ili prema Article 10(3) Direktive 2001/83/EC (hibridna aplikacija). U postupku razvoja hibridnih/generičkih lijekova glavna strategija je „kopiranje“ kvalitativnog i kvantitativnog sastava referentnog lijeka, a najveći izazov je dokazivanje terapijske ekvivalencije.Objectives Difference between drugs for dermal use and drugs that are intended for other routes of administration is that the selection of excipients depends on whether the goal is to achieve a systemic or local effect. In drugs for dermal use bioavailability of the active substance depends on its physical-chemical properties, pharmaceutical-technological features of the preparation, production process and way the drug is administered by a patient. The aim of this paper is to describe biopharmaceutical peculiarities of drugs for dermal use and contribute to the understanding of guidelines for preparation of documentation for marketing authorisation laid down by the European Medicines Agency (EMA), which are related to drug development, production, quality control and stability testing. Materials and methods Literature search was done by topic and subject of research, authors and journals, from general to specialized articles and guidelines relevant for the problem of the final paper. During literature search and collection of relevant data, on-line databases were used (PubMed, ScienceDirect, medicinal products database of the Agency for Medicinal Products and Medical Devices). Results Over the past 60 years topical delivery of drugs has become much more widely understood and much more is now known about the disposition of drugs in the skin. Today, different dermatological vehicles that are tailored to patients’ needs are produced. Additionally, better is appreciated how the formulation may affect rates of drug delivery, and ultimately, its efficacy and safety. On the market numerous systems for dermal use are available, but they are restricted to specific active substances. Further progress in the development of drugs for dermal use depends on the possibilities of overcoming the problems associated with the permeability of the skin and the skin irritation caused by the active substance and/or excipients. The development of new technologies that would increase the permeability and reduce skin irritation, would allow the use of hydrophilic molecules, macromolecules and conventional active ingredients for new therapeutic indications in preparations for dermal use. Conclusion Drugs for dermal use are big challenge for drug manufacturers due to the skin barrier whose primary role is to protect the body from external influences. Therefore, knowledge of the structure and function of the skin is crucial in the development of drugs for dermal use. Development of these drugs is based on the adjustment of transmission through the skin by choosing appropriate vehicles and on the use of modern pharmaceutical forms for dermal use (nanosystems). Application for granting marketing authorisation for drugs for dermal use in EU can be submitted according to Article 8(3) of Directive 2001/83/EC (full dossier) or according to Article 10(3) of Directive 2001/83/EC (hybrid application). In generic drug development, main strategy is „copying“ of qualitative and quantitative composition of reference product and greatest challenge is demonstration of therapeutic equivalence

Composition and Morphology Investigation of NiCoB Nanoparticles, as-Prepared and Coated by SiO2

NiCoB nanoparticles, as-prepared and coated by SiO2, were synthesized by chemical reduction of metallic salts. The as-prepared samples were additionally annealed at 650 for 1 hour in argon atmos-phere. All samples (the as-prepared and annealed ones) were investigated by X-ray powder diffraction (XRD), field emission scanning electron microscopy (FE-SEM) and energy dispersive spectroscopy (EDS). According to FE-SEM observations each of the investigated samples was composed of nearly spherical nanoparticles with average dimension smaller than 30 nm. Also, tendency of formation of irregular agglomerates was present in both cases of the as-prepared and annealed investigated samples. XRD measurements of as-prepared samples revealed fully amorphous structure. In case of annealed samples, crystallization of different phases was confirmed (as a consequence of heat treatment at 650 C). The existence of these phases was also observed from FE-SEM micrographs as different morphologies present in the samples.(doi: 10.5562/cca2234

Verification of automated latex-enhanced particle immunoturbidimetric D-Dimer assays on different analytical platforms and comparability of test results

Introduction: The aim of the study was the analytical verification of automated latex-enhanced particle immunoturbidimetric (LPIA) D-Dimer assay INNOVANCE D-dimer on Sysmex CS-5100 and Atellica COAG 360 analysers, and HemosIL D-dimer HS500 on ACL TOP 550, as well as the comparison with the enzyme-linked immunofluorescent assay (ELFA) on the miniVidas analyser. Materials and methods: Verification included assessment of within-run and between-run precision, bias, measurement uncertainty (MU), verification of the cut-off, method comparison between all assessed assays, and the reference commercial ELFA VIDAS D-Dimer Exclusion II. Results: Within-run coefficients of variations (CVs) ranged from 1.6% (Atellica COAG 360) to 7.9% (ACL TOP 550), while between-run CVs ranged from 1.7% (Sysmex CS-5100) to 6.9% (Atellica COAG 360). Spearman’s rank correlation coefficients were > 0.99 between LPIAs and ≥ 0.93 when comparing ELFA with LPIA. Passing-Bablok regression analysis yielded constant and proportional difference for comparison of ACL TOP 550 with both Sysmex CS-5100 and Atellica COAG360, and for miniVidas with Atellica COAG360. Small proportional difference was found between miniVidas and both Sysmex CS-5100 and ACL TOP 550. Calculated MUs using D-dimer HS 500 calibrator were 12.6% (Sysmex CS-5100) and 15.6% (Atellica COAG 360), while with INNOVANCE D-dimer calibrator 12.0% (Sysmex CS-5100), 10.0% (Atellica COAG 360) and 28.1% (ACL TOP 550). Excellent agreement of results was obtained, with occasional discrepancies near the cut-off. The cut-off (0.5 mg/L FEU) was confirmed. Conclusions: The obtained results prove satisfactory analytical performance of LPIAs, their high comparability and almost equal discriminatory characteristics, suggesting them as a valid alternative to ELFA

Diet Quality in Elderly Nursing Home Residents Evaluated by Diet Quality Index Revised (DQI-R)

The objective of this research was to evaluate diet quality in elderly nursing home residents and to point out the critical dietary components. The participants (277 females and 62 males) were recruited from all elderly nursing homes in Zagreb and each of elderly nursing homes was equally represented in this study. The age of subjects was ranging from 61 to 93 years; most of the females (53.4%) and males (53.2%) were between 70 and 80 years old. The dietary data from the multi pass 24-hour recall were used to compute the Diet Quality Index Revised (DQI-R). DQI-R is an instrument that provides a summary assessment of a diet’s overall healthfulness and is based on ten different aspects, including recommendations for both nutrient and food types. Pearson correlation analysis was used to compare the total DQI-R score with dietetic parameters and t-test was calculated between mean values of all the components of DQI-R as well as for total DQI-R score for men and women. The mean DQI-R score for the 339 sample was 62.1±11.7. The biggest number of participants satisfied recommendations about dietary cholesterol intake (88.5 % of participants) and dietary moderation score (71.1% of participants) but nobody satisfied recommendation about dietary diversity score. Only 3.2% of subjects had an adequate calcium intake (6.5% of male participants and only 2.5% of female participants). Recommended servings of fruit intake were satisfied by 19.8% of population, 30.4% satisfied vegetables recommendations and 38.6% recommendations for grains. According to DQI-R, beside positive dietary habits regarding dietary moderation and dietary cholesterol intake the population of elderly nursing home residents in the capital of Croatia needs improvement in other dietary habits in order to enhance successful aging

Prvi bolnički registar bolesnika s multiplom sklerozom u Hrvatskoj

Th e fi rst hospital-based registry of patients with multiple sclerosis (MS) was established at the University Department of Neurology, Sestre milosrdnice University Hospital Centre, Zagreb, Croatia, in 2014. Th e aim of the registry was to continuously provide data on the number of hospital-managed MS patients, patterns of disease progression, predictors of disability progression, changes in lifespan and long-term outcomes. Relevant medical data included age and gender of MS patients, family history of MS, data on previous immunization, disease course, Expanded Disability Status Scale (EDSS) score, cerebral magnetic resonance imaging (MRI) lesion load quantification, and cerebrospinal fl uid analysis. Lifestyle habits in MS patients including smoking and alcohol consumption were also analyzed. All data were obtained from primary medical records between January 1, 2014 and January 1, 2015, and entered into the database. Data were evaluated retrospectively according to age and gender diff erences. Results showed that the majority of patients enrolled in the registry had the remitting relapsing course of disease, with low EDSS score indicating no disability or minimal disability. Cerebrospinal fl uid analysis showed that oligoclonal bands were present in the majority of MS patients, with aff ected blood-brain-barrier permeability. According to the remitting relapsing course of the disease, cerebral MRI quantitative analysis demonstrated a signifi cant lesion load in the majority of patients. When stratifi ed by lifestyle habits, smokers and alcohol consumers were more prevalent among male patients. Our hospital-based registry might be considered as a prototype for the national MS registry and should be improved for reliable statistical analysis.Prvi bolnički registar bolesnika s multiplom sklerozom (MS) u Hrvatskoj uspostavljen je 2014. godine u Klinici za neurologiju, Klinički bolnički centar “Sestre milosrdnice”, Zagreb. Cilj registra je kontinuirano prikupljanje podataka o broju hospitalno obrađenih bolesnika s MS, oblicima bolesti, prediktorima napredovanja onesposobljenosti, životnim navikama i dugoročnim ishodima za bolesnike. Relevantni medicinski podaci u registru MS uključivali su procjenu dobi i spola bolesnika, obiteljsku anamnezu na MS, podatke o prethodnim imunizacijama, tijeku bolesti, procjenu funkcionalne onesposobljenosti, kvantitativni prikaz lezija mozga magnetskom rezonancijom (MRI) i analizu cerebrospinalnog likvora. Nadalje, analizirane su životne navike bolesnika uključujući pušenje i konzumaciju alkohola. Svi podaci iz bolničke medicinske dokumentacije koji su prikupljeni od 1. siječnja 2014. godine do 1. siječnja 2015. godine upisani su u bazu podataka. Retrospektivno smo analizirali podatke prema dobi i spolu. Relevantni rezultati su pokazali da većina registriranih bolesnika ima relapsno remitirajući oblik bolesti s niskim stupnjem funkcionalne onesposobljenosti. Analiza cerebrospinalnog likvora u većine bolesnika pokazala je pozitivne oligoklonske vrpce i poremećenu propusnost krvno-moždane barijere. Kvantitativna MRI analiza mozga pokazala je značajno opterećenje brojem cerebralnih lezija u većine bolesnika, što odgovara relapsno remitirajućem obliku bolesti. Podjela prema životnim navikama pokazala je da je bilo više pušača i konzumenata alkohola u skupini muških bolesnika. Naš bolnički registar može poslužiti kao prototip za uspostavljanje nacionalnog registra MS, ali ga treba pobolj šati kako bi se osigurali svi potrebni podaci za pouzdanu statističku analizu podataka

Reevaluation of von Willebrand disease diagnosis in a Croatian paediatric cohort combining bleeding scores, phenotypic laboratory assays and next generation sequencing: a pilot study

This study reevaluated von Willebrand disease (vWD) diagnosis in a Croatian paediatric cohort by combining bleeding scores (BS), phenotypic laboratory testing, and next-generation sequencing (NGS). A total of 25 children (11 males and 14 females, median age 10 years, from 2 to 17) previously diagnosed with vWD were included. BS were calculated using an online bleeding assessment tool. Phenotypic laboratory analyses included platelet count, platelet function analyser closure times, prothrombin time, activated partial thromboplastin time, von Willebrand factor antigen (vWF:Ag), vWF gain-of-function mutant glycoprotein Ib binding activity (vWF:GPIbM), vWF collagen binding activity (vWF:CBA), factor VIII activity (FVIII:C) and multimeric analysis. Next-generation sequencing covered regions of both vWF and FVIII genes and was performed on MiSeq (Illumina, San Diego, USA). Disease-associated variants identified in 15 patients comprised 11 distinct heterozygous vWF gene variants in 13 patients and one novel FVIII gene variant (p.Glu2085Lys) in two male siblings. Four vWF variants were novel (p.Gln499Pro, p.Asp1277Tyr, p.Asp1277His, p.Lys1491Glu). Three patients without distinctive variants had vWF:GPIbM between 30 and 50%. Patients with identified vWF gene variants had statistically significant lower values of vWF:GPIbM (P = 0.002), vWF:Ag (P = 0.007), vWF:CBA (P < 0.001) and FVIII:C (P = 0.002), compared to those without. Correlations between BS and phenotypic laboratory test results were not statistically significant for either of the tests. The applied diagnostic approach confirmed the diagnosis of vWD in 13 patients and mild haemophilia A in two. Limited utility of BS in the paediatric population was evidenced

Clinical significance of T315I ABL kinase domain mutation detection in patients resistant to imatinib mesylate therapy

Uvod: Kronična mijeloična leukemija (engl. chronic myeloid leukemia, CML) je mijeloproliferativna bolest koju karakterizira prisutnost fuzijskog gena bcr-abl i posljedično fuzijskog proteina bcr-abl. Iako je otkriće inhibitora tirozin-kinaze (engl. tyrosine kinase inhibitor, TKI), imatinib mesilata (IM) poboljšalo liječenje bolesnika oboljelih od CML, dio bolesnika razvija rezistenciju na lijek, što dovodi do povišene razine bcr-abl prijepisa. Jedan od mogućih razloga te rezistencije su mutacije u domeni ABL kinaze. Neke se mutacije mogu prevladati povećanjem doze lijeka ili primjenom nove generacije TKI. Jedina mutacija rezistentna na trenutno dostupne TKI jest T315I. Cilj ovog istraživanja bio je otkriti da li je prisutnost T315I kod bolesnika rezistentnih na liječenje imatinib mesilatom povezana s povećanom ili neprekidno visokom razinom bcr-abl prijepisa. Također, cilj je bio procijeniti moguću razliku u razini bcr-abl prijepisa kod bolesnika rezistentnih na liječenje imatinib mesilatom sa i bez T315I mutacije. Ispitanici i metode: U ispitivanje su bila uključena 24 bolesnika oboljela od CML s neodgovarajućim odgovorom na liječenje imatinib mesilatom. Provedena je kvantitativna lančana reakcija polimerazom u stvarnom vremenu (engl. real time quantitative polymesare chain reaction, RQ-PCR) prema protokolu udruženja Europa protiv raka (engl. Europe Against Cancer), a za dokazivanje mutacije T315I rabljena je metoda alel specifične oligonukleotidne PCR (engl. allele specific oligonucleotide PCR, ASO PCR). Rezultati: Kod 4 od 24 bolesnika dokazana je mutacija T315I (17%). Izrač u-nat je i medijan omjera bcr-abl/abl koji je iznosio 19% za bolesnike s T315I mutacijom i 13% za bolesnike bez mutacije, no razlika između tih dviju skupina nije bila statistički značajna (P = 0,394) Zaključak: Dokazivanje prisutnosti mutacije T315I ključno je u terapijskom pristupu bolesnicima s CML, budući daje liječenje izbora za nositelje te mutacije transplantacija matičnih stanica.Background: Chronic myeloid leukemia (CML) is a myeloproliferative disease characterized by the presence of bcr-abl fusion gene and consequently bcr-abl fusion protein. Although the discovery of tyrosine kinase inhibitor (TKI), imatinib mesylate (IM), improved the treatment of CML patients, a proportion of patients develop resistance to the drug resulting in increased bcr-abl level. One of possible reasons for resistance are the mutations in ABL kinase domain. Some mutations can be overcome by increasing the drug dose or by using the new generation of TKI. The only mutation resistant to currently available TKI is T315I. The aim of this study was to detect if the presence of T315I in patients resistant to imatinib mesylate therapy is associated with the increase or constantly high bcr-abl level. We also aimed to assess the possible difference in bcr-abl level in imatinib-resistant patients with and without T315I mutation. Materials and methods: The study included 24 CML patients with inadequate response to IM therapy. Real time quantitative PCR was performed according to Europe Against Cancer protocol and allele specific oligonucleotide PCR was used for T315I mutation detection. Results:T315I was detected in 4out of 24 patients (17%). Calculated median bcr-abl/abl levels were 19%forT315I positive and 13%forT315I negative patients, but the difference was not statistically significant (P = 0.394). Conclusions: T315I detection is essential in therapy approach for CML patients as the treatment of choice for T315I carriers is stem-cell transplantation

Clinical significance of T315I ABL kinase domain mutation detection in patients resistant to imatinib mesylate therapy

Uvod: Kronična mijeloična leukemija (engl. chronic myeloid leukemia, CML) je mijeloproliferativna bolest koju karakterizira prisutnost fuzijskog gena bcr-abl i posljedično fuzijskog proteina bcr-abl. Iako je otkriće inhibitora tirozin-kinaze (engl. tyrosine kinase inhibitor, TKI), imatinib mesilata (IM) poboljšalo liječenje bolesnika oboljelih od CML, dio bolesnika razvija rezistenciju na lijek, što dovodi do povišene razine bcr-abl prijepisa. Jedan od mogućih razloga te rezistencije su mutacije u domeni ABL kinaze. Neke se mutacije mogu prevladati povećanjem doze lijeka ili primjenom nove generacije TKI. Jedina mutacija rezistentna na trenutno dostupne TKI jest T315I. Cilj ovog istraživanja bio je otkriti da li je prisutnost T315I kod bolesnika rezistentnih na liječenje imatinib mesilatom povezana s povećanom ili neprekidno visokom razinom bcr-abl prijepisa. Također, cilj je bio procijeniti moguću razliku u razini bcr-abl prijepisa kod bolesnika rezistentnih na liječenje imatinib mesilatom sa i bez T315I mutacije. Ispitanici i metode: U ispitivanje su bila uključena 24 bolesnika oboljela od CML s neodgovarajućim odgovorom na liječenje imatinib mesilatom. Provedena je kvantitativna lančana reakcija polimerazom u stvarnom vremenu (engl. real time quantitative polymesare chain reaction, RQ-PCR) prema protokolu udruženja Europa protiv raka (engl. Europe Against Cancer), a za dokazivanje mutacije T315I rabljena je metoda alel specifične oligonukleotidne PCR (engl. allele specific oligonucleotide PCR, ASO PCR). Rezultati: Kod 4 od 24 bolesnika dokazana je mutacija T315I (17%). Izrač u-nat je i medijan omjera bcr-abl/abl koji je iznosio 19% za bolesnike s T315I mutacijom i 13% za bolesnike bez mutacije, no razlika između tih dviju skupina nije bila statistički značajna (P = 0,394) Zaključak: Dokazivanje prisutnosti mutacije T315I ključno je u terapijskom pristupu bolesnicima s CML, budući daje liječenje izbora za nositelje te mutacije transplantacija matičnih stanica.Background: Chronic myeloid leukemia (CML) is a myeloproliferative disease characterized by the presence of bcr-abl fusion gene and consequently bcr-abl fusion protein. Although the discovery of tyrosine kinase inhibitor (TKI), imatinib mesylate (IM), improved the treatment of CML patients, a proportion of patients develop resistance to the drug resulting in increased bcr-abl level. One of possible reasons for resistance are the mutations in ABL kinase domain. Some mutations can be overcome by increasing the drug dose or by using the new generation of TKI. The only mutation resistant to currently available TKI is T315I. The aim of this study was to detect if the presence of T315I in patients resistant to imatinib mesylate therapy is associated with the increase or constantly high bcr-abl level. We also aimed to assess the possible difference in bcr-abl level in imatinib-resistant patients with and without T315I mutation. Materials and methods: The study included 24 CML patients with inadequate response to IM therapy. Real time quantitative PCR was performed according to Europe Against Cancer protocol and allele specific oligonucleotide PCR was used for T315I mutation detection. Results:T315I was detected in 4out of 24 patients (17%). Calculated median bcr-abl/abl levels were 19%forT315I positive and 13%forT315I negative patients, but the difference was not statistically significant (P = 0.394). Conclusions: T315I detection is essential in therapy approach for CML patients as the treatment of choice for T315I carriers is stem-cell transplantation

Serum chitotriosidase: a circulating biomarker in polycythemia vera

Objectives: Serum chitotriosidase activity (CHIT1) is a biomarker of macrophage activation with an important role in inflammation-induced tissue remodeling and fibrosis. Macrophages have been described to play a crucial role in regulating pathological erythropoiesis in polycythemia vera (PV). The aim of this study was to evaluate CHIT1 in patients diagnosed with Philadelphia-negative myeloproliferative neoplasms (MPNs). ----- Methods: Using fluorometric assay, we measured CHIT1 in 28 PV, 27 essential thrombocythemia (ET), 17 primary myelofibrosis (PMF), 19 patients with secondary myelofibrosis and in 25 healthy controls. ----- Results: CHIT1 was significantly higher in PV (p < .001) and post-PV myelofibrosis (MF) transformation (post-PV MF) (p = .020), but not in ET (p = .080), post-ET MF transformation (p = .086), and PMF patients (p = .287), when compared to healthy controls. CHIT1 in PV was positively correlated with hemoglobin (p = .026), hematocrit (p = .012), absolute basophil count (p = .030) and the presence of reticulin fibrosis in the bone marrow (p = .023). ----- Discussion: A positive correlation between CHIT1 and these distinct laboratory PV features might imply macrophages closely related to clonal erythropoiesis as cells of CHIT1 origin. In addition, a positive association between CHIT1 and reticulin fibrosis might indicate its potential role in PV progression. ----- Conclusion: CHIT1 might be considered as a circulating biomarker in PV. Additional studies are needed to clarify the role of CHIT1 in promoting disease progression and bone marrow fibrosis in PV