CEFYBO   02669
CENTRO DE ESTUDIOS FARMACOLOGICOS Y BOTANICOS
Unidad Ejecutora - UE
artículos
Título:
Two recombinant human interferon-beta 1a pharmaceutical preparations produce a similar transcriptional response determined using whole genome microarrays analysis
Autor/es:
STERIN-PRYNC A; YANKILEVICH PATRICIO; BARRERO P; BELLO RICARDO; MARANGUNICH LAURA; VIDAL ALEJANDRO; CRISCUOLO MARCELO; KAUFFMAN MARCELO; DIEZ ROBERTO
Revista:
INTERNATIONAL JOURNAL OF CLINICAL PHARMACOLOGY AND THERAPEUTICS
Referencias:
Año: 2008 vol. 46 p. 64 - 71
ISSN:
0946-1965
Resumen:
Ab stract. Objectives: Re com bi nant human in ter feron-beta (IFN-b) is a well-established treatment for multiple sclerosis (MS). The reg u la tory pro cess for mar ket ing au tho riza tion of biosimilars is cur rently un der de bate in cer tain coun tries. In the EU, EMEA has clearly de fined the pro cess in clud ing overarch ing and prod uct-spe cific guide lines which includes clinical testing. Biosimilarity needs to be based on com pa ra bil ity cri te ria, including at least molecular characterization, bio log ical activity relevant for the ther apeu ticef fect and relative bioavailability (“bioequivalence”). In the case of such com plex diseases as MS, where the ef fect of treat ment is not so directly mea sur able, in vi tro tools can pro vide ad di tional data to sup port com pa ra bil ity. Genomic microarrays as says might be use ful to com pare multisource biopharmaceuticals. The aim of the pres ent study was to com pare the pharmacodynamic genomic ef fects (in terms of transcriptional reg u la tion) of two recom bi nant hu man IFN-b-1a prep a ra tions on lymphocytes of multiple sclerosis patients using a whole ge nome microarray as say. Methods: We per formed an ex vivo whole ge nome ex pres sion pro fil ing of the ef fect of two prepa ra tions of IFN-b1a on non-ad her ent mononuclears from five re laps ing-re mit ting MS patients analyzing microarrays (CodeLink™Hu man Whole Ge nome). Patients blood was drawn, PBMCs iso lated and cul tured in three dif fer ent con di tions: cul ture me dium (control), 1,000 U/ml of IFN-b1a (BLASTOFERON™, Bio Sidus) and 1,000 U/ml of IFN-b1a (REBIF™, Serono) RNA was puri fied from non-ad her ent cells (mostly lympho cytes), am pli fied and hy brid ized. Raw data were gen er ated by CodeLink™ pro prietary software. Data normalization, quality control and anal y sis of dif fer en tial gene expres sion be tween treat ments were done using linear model for microarray data. Func tional annotation analysis of IFN-b1a MS treatment trascription was done us ing DAVID. Results: Out of the approx i mately 45,000 human se quences ex am ined, no ev i dence of differential reglation was found when both treatments were com pared (minimum ad justed p-value > 0.999). The IFN-b1a ef fect differen tially regulated the expression of 868 genes. The expression of standard markers such as GTP cyclohidrolase, MxA, and OAS isoenzymes A and B changed as a consequence of the ac tion of IFN-b1a. Conclusions: This exhaustive and highly sensitive assay did not show dif fer ences in the genomic expression profile of these two products under the assayed experimen tal condi tions. These re sults sug gest that this technology might be use ful for the initial comparison of biosimilars, being part of a compre hensive comparabil ity program that includes clinical testing.Objectives: Re com bi nant human in ter feron-beta (IFN-b) is a well-established treatment for multiple sclerosis (MS). The reg u la tory pro cess for mar ket ing au tho riza tion of biosimilars is cur rently un der de bate in cer tain coun tries. In the EU, EMEA has clearly de fined the pro cess in clud ing overarch ing and prod uct-spe cific guide lines which includes clinical testing. Biosimilarity needs to be based on com pa ra bil ity cri te ria, including at least molecular characterization, bio log ical activity relevant for the ther apeu ticef fect and relative bioavailability (“bioequivalence”). In the case of such com plex diseases as MS, where the ef fect of treat ment is not so directly mea sur able, in vi tro tools can pro vide ad di tional data to sup port com pa ra bil ity. Genomic microarrays as says might be use ful to com pare multisource biopharmaceuticals. The aim of the pres ent study was to com pare the pharmacodynamic genomic ef fects (in terms of transcriptional reg u la tion) of two recom bi nant hu man IFN-b-1a prep a ra tions on lymphocytes of multiple sclerosis patients using a whole ge nome microarray as say. Methods: We per formed an ex vivo whole ge nome ex pres sion pro fil ing of the ef fect of two prepa ra tions of IFN-b1a on non-ad her ent mononuclears from five re laps ing-re mit ting MS patients analyzing microarrays (CodeLink™Hu man Whole Ge nome). Patients blood was drawn, PBMCs iso lated and cul tured in three dif fer ent con di tions: cul ture me dium (control), 1,000 U/ml of IFN-b1a (BLASTOFERON™, Bio Sidus) and 1,000 U/ml of IFN-b1a (REBIF™, Serono) RNA was puri fied from non-ad her ent cells (mostly lympho cytes), am pli fied and hy brid ized. Raw data were gen er ated by CodeLink™ pro prietary software. Data normalization, quality control and anal y sis of dif fer en tial gene expres sion be tween treat ments were done using linear model for microarray data. Func tional annotation analysis of IFN-b1a MS treatment trascription was done us ing DAVID. Results: Out of the approx i mately 45,000 human se quences ex am ined, no ev i dence of differential reglation was found when both treatments were com pared (minimum ad justed p-value > 0.999). The IFN-b1a ef fect differen tially regulated the expression of 868 genes. The expression of standard markers such as GTP cyclohidrolase, MxA, and OAS isoenzymes A and B changed as a consequence of the ac tion of IFN-b1a. Conclusions: This exhaustive and highly sensitive assay did not show dif fer ences in the genomic expression profile of these two products under the assayed experimen tal condi tions. These re sults sug gest that this technology might be use ful for the initial comparison of biosimilars, being part of a compre hensive comparabil ity program that includes clinical testing.