RESUMO
Peptide KVPLITVSKAK was selected to design a synthetic ligand for affinity chromatography purification of recombinant human follicle stimulating hormone (rhFSH), based on the interaction of the hormone with the exoloop 3 of its receptor. The peptide was acetylated to improve its stability to degradation by exopeptidases. A cysteine was incorporated at the C-termini to facilitate its immobilization to the chromatographic activated SulfoLink agarose resin. A sample of crude rhFSH was loaded to the affinity column, using 20 mM sodium phosphate, 0.5 mM methionine, and pH 5.6 and 7.2 as adsorption and elution buffers, respectively. The dynamic capacity of the matrix was 54.6 mg rhFSH/mL matrix and the purity 94%. The percentage of oxidized rhFSH was 3.4%, and that of the free subunits was 1.2%, both in the range established by the European Pharmacopeia, as also were the sialic acid content and the isoforms profile.
Assuntos
Cromatografia de Afinidade/métodos , Hormônio Foliculoestimulante Humano/isolamento & purificação , Peptídeos/metabolismo , Proteínas Recombinantes/isolamento & purificação , Acetilação , Animais , Células CHO , Cricetulus , Hormônio Foliculoestimulante Humano/química , Hormônio Foliculoestimulante Humano/metabolismo , Humanos , Proteínas Imobilizadas/síntese química , Proteínas Imobilizadas/metabolismo , Peptídeos/síntese química , Estabilidade Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismoRESUMO
BACKGROUND: Triptorelin is an established treatment for central precocious puberty (CPP) as 1- and 3-month formulations. The current triptorelin 22.5 mg 6-month formulation is approved for prostate cancer therapy. This is the first study in patients with CPP. METHODS: The efficacy and safety of the triptorelin 6-month formulation in CPP were investigated. The primary objective was to evaluate the efficacy in achieving luteinizing hormone (LH) suppression to pre-pubertal levels at month 6. This was an international, non-comparative phase III study over 48 weeks. Eighteen medical centers in the US, Chile and Mexico participated. Forty-four treatment naïve patients (39 girls and five boys) aged at treatment start 2-8 years for girls and 2-9 years for boys with an advancement of bone age over chronological age ≥1 year were to be included. Triptorelin was administered im twice at an interval of 24 weeks. LH, follicle stimulating hormone (FSH) (basal and stimulated), estradiol (girls), testosterone (boys), auxological parameters, clinical signs of puberty and safety were assessed. RESULTS: Forty-one patients (93.2%) showed pre-pubertal LH levels (stimulated LH ≤5 IU/L) at month 6 and maintained LH suppression through month 12. The percentage of patients with LH suppression exceeded 93% at each time point and reached 97.7% at month 12. No unexpected drug-related adverse events were reported. CONCLUSIONS: The triptorelin 6-month formulation was safe and effective in suppressing the pituitary-gonadal axis in children with CPP. The extended injection interval may improve compliance and increase comfort in the management of CPP.
Assuntos
Hormônio Liberador de Gonadotropina/agonistas , Hormônio Luteinizante/antagonistas & inibidores , Puberdade Precoce/tratamento farmacológico , Substâncias para o Controle da Reprodução/administração & dosagem , Pamoato de Triptorrelina/administração & dosagem , Biomarcadores/sangue , Criança , Desenvolvimento Infantil/efeitos dos fármacos , Pré-Escolar , Chile , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/efeitos adversos , Preparações de Ação Retardada/uso terapêutico , Esquema de Medicação , Estradiol/sangue , Estradiol/química , Estradiol/metabolismo , Feminino , Hormônio Foliculoestimulante Humano/antagonistas & inibidores , Hormônio Foliculoestimulante Humano/sangue , Hormônio Foliculoestimulante Humano/metabolismo , Humanos , Hormônio Luteinizante/sangue , Hormônio Luteinizante/metabolismo , Masculino , México , Osteogênese/efeitos dos fármacos , Puberdade Precoce/sangue , Puberdade Precoce/metabolismo , Substâncias para o Controle da Reprodução/efeitos adversos , Substâncias para o Controle da Reprodução/uso terapêutico , Testosterona/antagonistas & inibidores , Testosterona/sangue , Testosterona/metabolismo , Pamoato de Triptorrelina/efeitos adversos , Pamoato de Triptorrelina/uso terapêutico , Estados UnidosRESUMO
OBJECTIVE: To describe the management of a patient with a pituitary adenoma secreting follicle-stimulating hormone (FSH) associated with ovarian hyperstimulation who was treated with a gonadotropin-releasing hormone (GnRH) antagonist. DESIGN: Case report. SETTING: University teaching hospital. PATIENT(S): A woman of reproductive age with secondary amenorrhea and ovarian hyperstimulation due to a pituitary adenoma secreting FSH, which persisted after transsphenoidal surgery. INTERVENTION(S): Clinical treatment with a GnRH antagonist. MAIN OUTCOME MEASURE(S): A decrease in serum estradiol levels. RESULT(S): During the treatment period, ovarian hyperstimulation decreased as shown by a reduction in estradiol levels and an improvement in the patient's clinical condition and in the ultrasonography parameters. CONCLUSION(S): The GnRH antagonist was found to be effective for the short-term treatment of ovarian hyperstimulation secondary to a pituitary adenoma secreting FSH, thus representing a therapeutic option that should be taken into consideration in such cases.
Assuntos
Adenoma/tratamento farmacológico , Hormônio Foliculoestimulante Humano/metabolismo , Hormônio Liberador de Gonadotropina/análogos & derivados , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Síndrome de Hiperestimulação Ovariana/tratamento farmacológico , Neoplasias Hipofisárias/tratamento farmacológico , Adenoma/complicações , Adenoma/metabolismo , Adenoma/cirurgia , Adulto , Feminino , Hormônio Liberador de Gonadotropina/administração & dosagem , Antagonistas de Hormônios/administração & dosagem , Humanos , Síndrome de Hiperestimulação Ovariana/etiologia , Neoplasias Hipofisárias/complicações , Neoplasias Hipofisárias/metabolismo , Neoplasias Hipofisárias/cirurgia , Resultado do TratamentoRESUMO
The effects of human FSH glycoforms on mouse follicle development and function in vitro were analysed, and an attempt was made to relate markers of follicular maturation to the expression of immunolocalized connexin (Cx) 43 and Cx26-based gap junctions. Three FSH fractions comprising discrete pI ranges [7.10-5.99 (pool I), pI 5.62-4.95 (pool II) and <3.75 (pool III)] were studied. Pool I produced the strongest effect on preantral granulosa cell proliferation and oestradiol production, and was highly effective for stimulating antral formation; this isoform also evoked a peripheral distribution of Cx43-containing gap junctions. Pool II was effective in promoting preantral granulosa cell proliferation but required higher FSH doses. This particular isoform provoked a more central distribution of Cx43-containing gap junctions, which was associated with a lower oestradiol production and less effective antral formation. Pool III was the least active for all markers of follicle development, and this was associated with minimal induction of Cx43-based gap junctions. The effects of the three FSH isoform pools on Cx26 expression were similar. The pattern of differences strongly suggests that FSH isoforms have complementary and specific actions on developing follicles, and that a shifting stage specific balance of isoforms is required for optimal follicle development.