Streszczenie -

Streszczenie
Zanieczyszczenia i produkty degradacji fumaranu bisoprololu
w substancji czynnej oraz w produkcie leczniczym
Praca
y Bisocard®
podsumowanie przeprowadzonych prac.
1.
zy substancji czynnej oraz przedstawiono
analiz, zmniejszenie zu
otrzymano na Acquity® UPLC, BEH C18, Waters, 100 x 2,1 mm, 1,7 µm.
przedstawiono w Publikacji I.
2.
problemy oznaczania
metody badania
naj
yniki
w fumaranie bisoprololu. Odtworzenie
tym opracowano i zwalidowano
-FID do oznaczania epichlorohydryny oraz
HPLC/UV
do kontrolowania poziomu epoksybisoprololu. Analiza substancji czynnej wytwarzanej w ICN
Polfa Rzeszów S.A.
rutynowej kontroli.
3.
Opracowano
-FID Headspace do równoczesnego oznaczania chlorku metylenu,
toluenu, propan-2-olu, 2-izopropoksyetanolu i benzenu.
,w
trzech kolejnych seriach produkcyjnych substancji czynnej, pozwalaj
ich rutynowej
kontroli.
4.
e
GC-FID Headspace
,
1.
Poznano
badanej substancji
u fenylu). Walidacja
izopropyloaminy, wyznaczona w
nie od jej rutynowej kontroli.
. W badaniach obserwowano
mechanizmów typu SN1 i SN2. Dla obu reakcji wyznaczono parametry termodynamiczne.
bardzo stabilne. Otrzymane wyniki opisano w Publikacji II. W
badaniach preformulacyjnych un
i nadmiaru wody.
2.
Badano przebieg reakcji Maillarda i przegrupowania Amadoriego
bisoprololem a cukrami
na szybk
. Wyniki wykorzystano
przemiany. Zidentyfikowano
produktu Maillarda, produktu
przegrupowania Amadoriego i produktu
ego reakcji (N-formylobisoprololu) przy pomocy techniki
LC/MS2. Wyniki opisano w Publikacji III.
3.
leki,
, poddano
przyspieszonym
produktów oraz wyznaczono tempo przyrostu
poszczególnych degradantów.
4.
procesów
-
Próby poddano
Otrzymane wyniki wykorzystano do wytypowania formulacji,
która zapewni
produktów z fumaranem bisoprololu. Na podstawie otrzymanych
wyników wybrano najkorzystniejszy proces powlekania
.
Abstract
Impurities and degradation products of bisoprolol fumarate
in the active substance and in the drug product
This doctoral dissertation was performed for ICN Polfa Rzeszów S.A. The main scope of our studies was to
resolve analytical problems that prevented the registration of the active substance in the EU member states and to
improve the stability of Bisocard® drug products. Current ICH, EMA and FDA guidelines were followed. A
short summary was presented below.
Part One:
1.
Characterisation of impurities arising during the synthesis of the drug substance and degradants arising
during storage of the drug product was performed. A new VHPLC method for the determination of related
substances in bisoprolol fumarate was developed. Analysis time, consumption of organic solvents, and
formation of chemical waste was significantly reduced. The proposed theoretical model enabled the
comparison of resolution power of chromatographic columns filled with totally porous as well as core-shell
adsorbents. Obtained results were summarized in Publication I.
2.
Analytical problems with the determination of genotoxic impurities in bisoprolol fumarate were solved. New
GC-FID method and new HPLC/UV method were developed to control the concentration of epichlorohydrin
and epoxybisoprolol, respectively. Both methods were fully validated according to ICH guidelines. The
active substance manufactured at ICN Polfa Rzeszów S.A. is free from genotoxic impruities.
3.
A new GC-FID Headspace method was developed and validated for the simultaneous determination of
several organic volatile impurities (methylene chloride, toluene, propan-2-ol, 2-isopropoxyethanol, benzene).
Class 1 solvent was not detected. Only trace amounts of other mentioned compounds were found in the final
active substance. The routine analysis of the residual solvents in bisoprolol fumarate was skipped based on
ICH guidelines.
4.
Difficulties in the determination of isopropylamine with GC-FID Headspace technique were found. That’s
why the impurity was controlled by a new HPLC/UV method with a pre-column derivatization with phenyl
isocyanate. The content of isopropylamine in bisoprolol fumarate was lower than 40 ppm in each of the three
consecutive production scale batches, thus the routine control was skipped.
Part Two:
5.
Kinetics of degradation of bisoprolol fumarate in aqueous acidic solutions was studied. Hydrolysis of the
benzyl ether linkage occurred according to two competitive mechanisms (SN1 and SN2). Activation
parameters for both reactions were calculated. Other functional groups of bisoprolol remained unchanged.
Obtained results were presented in Publication II. In the pre-formulation studies both acidic environment and
excess of water were avoided.
6.
Formulation incompatibilities between secondary amine of bisoprolol and reducing carbohydrates, which
resulted in the non-enzymatic browning reactions, were thoroughly studied. Such pharmaceutical systems are
usually unstable and improve the degradation of the drug substance. Results of experiments in low-moisture
alkaline liquid mixtures were used to present the reaction mechanism. The influence of several diluents
and lubricants on the pace of the non-enzymatic browning reaction was tested. Structures of appropriate
glycosylamines, Amadori rearrangement products and N-formylbisoprolol (final reaction product) were
confirmed by LC/MS2 studies. Obtained results were summarized in Publication III.
7.
Marketed drug products, containing bisoprolol fumarate, were subjected to the accelerated stability studies.
Obtained impurity profiles and pace of the formation of individual degradants were carefully considered.
The most stable marketed formulation of the drug product was recognised.
8.
Several laboratory scale batches with various qualitative compositions and different film-coating processes
were manufactured. The batches were packed into various immediate packages and subjected to the
accelerated stability studies. The influence of a diluent type on the impurity profile was assessed. Obtained
results were utilized to present the most stable formulation, the most advantageous film-coating process
and the most effective immediate package for the drug product.