RESUMEN
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder which primarily affects the gastrointestinal and nervous systems. This disease is caused by mutations in the nuclear TYMP gene, which encodes for thymidine phosphorylase, an enzyme required for the normal metabolism of deoxynucleosides, thymidine, and deoxyuridine. The subsequent elevated systemic concentrations of deoxynucleosides lead to increased intracellular concentrations of their corresponding triphosphates, and ultimately mitochondrial failure due to progressive accumulation of mitochondrial DNA (mtDNA) defects and mtDNA depletion. Currently, there are no treatments for MNGIE where effectiveness has been evidenced in clinical trials. This Phase 2, multi-centre, multiple dose, open label trial without a control will investigate the application of erythrocyte-encapsulated thymidine phosphorylase (EE-TP) as an enzyme replacement therapy for MNGIE. Three EE-TP dose levels are planned with patients receiving the dose level that achieves metabolic correction. The study duration is 31 months, comprising 28 days of screening, 90 days of run-in, 24 months of treatment and 90 days of post-dose follow-up. The primary objectives are to determine the safety, tolerability, pharmacodynamics, and efficacy of multiple doses of EE-TP. The secondary objectives are to assess EE-TP immunogenicity after multiple dose administrations and changes in clinical assessments, and the pharmacodynamics effect of EE-TP on clinical assessments.
RESUMEN
We have previously shown that intravenous (i.v.) treatment with sterically stabilized nano-liposomes (NSSL) actively remote-loaded with the glucocorticoid (GC) methylprednisolone hemisuccinate (NSSL-MPS) or betamethasone hemisuccinate (NSSL-BMS) significantly decreased severity of adjuvant arthritis in Lewis rats (a model of human rheumatoid arthritis) throughout all disease stages. Here, we compared i.v. or subcutaneous (s.c.) weekly treatment with each of the two NSSL-GC to weekly or daily treatment with the free drugs or with the TNF-α antagonists Infliximab and Etanercept. Therapeutic efficacy and effects on the profile of pro-inflammatory (IL-6, TNF-α, and INF-γ) and anti-inflammatory (IL-10 and TGF-ß) cytokines in rat sera and splenocyte tissue culture supernatants were compared to those of the liposomal and free drugs. Both s.c. and i.v. NSSL-GC suppressed arthritis significantly, compared to higher doses of the free drugs or to TNF-α antagonists. NSSL-GC also suppressed the secretion of pro-inflammatory cytokines, but did not change the levels of TGF- ß. The highly efficacious anti-inflammatory therapeutic feature of these nano-drugs makes them candidates for treatment of human rheumatoid arthritis.
Asunto(s)
Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Citocinas/sangre , Portadores de Fármacos/química , Glucocorticoides/uso terapéutico , Nanopartículas/química , Animales , Antiinflamatorios/administración & dosificación , Antiinflamatorios/sangre , Artritis Experimental/inmunología , Betametasona/administración & dosificación , Betametasona/uso terapéutico , Células Cultivadas , Esquema de Medicación , Femenino , Glucocorticoides/administración & dosificación , Glucocorticoides/sangre , Inyecciones Intravenosas , Inyecciones Subcutáneas , Liposomas , Hemisuccinato de Metilprednisolona/administración & dosificación , Hemisuccinato de Metilprednisolona/uso terapéutico , Ratones , Ratones Endogámicos , Ratas , Ratas Endogámicas Lew , Bazo/citología , Bazo/efectos de los fármacos , Bazo/inmunología , Resultado del TratamientoRESUMEN
Hemophilia A is a rare X-linked bleeding disorder caused by lack or dysfunction of coagulation factor VIII (FVIII). Hemophilia A is treated with replacement therapy, but frequent injections of the missing FVIII often lead to the formation of inhibitory antibodies. Patients who develop high levels of inhibitors must be treated with bypassing agents such as activated FVII (FVIIa). Both FVIII and FVIIa have short half-lives and require multiple injections. Long-acting forms of these proteins would therefore reduce the frequency of injections, improve patient compliance and reduce complications. In this article we present a new platform technology that produces long-acting forms of FVIII and FVIIa and improves the efficacy of hemophilia treatment. This technology is based on the binding of proteins/peptides to the outer surface of PEGylated liposomes (PEGLip). Binding is dependent on an amino acid consensus sequence within the proteins and is highly specific. At the same time, binding is non-covalent and does not require any modification of the therapeutic agent or its production process. Association of proteins with PEGLip results in substantial enhancements in their pharmacodynamic properties following administration. These improvements seem to arise from the association of formulated proteins with platelets prior to induction of coagulation.
Asunto(s)
Sistemas de Liberación de Medicamentos/métodos , Factor VIII/administración & dosificación , Factor VIIa/administración & dosificación , Hemofilia A/tratamiento farmacológico , Secuencia de Aminoácidos , Animales , Química Farmacéutica , Secuencia de Consenso , Preparaciones de Acción Retardada , Modelos Animales de Enfermedad , Factor VIII/genética , Factor VIIa/genética , Hemofilia A/sangre , Humanos , Liposomas/química , Ratones , Nanomedicina , Polietilenglicoles/químicaRESUMEN
IMPORTANCE OF THE FIELD: Improving the pharmacodynamics of protein drugs has the potential to improve the care and the quality of life of patients suffering from a variety of diseases. AREAS COVERED IN THIS REVIEW: Four approaches to improve protein drugs are described: PEGylation, amino acid substitution, fusion to carrier proteins and encapsulation. A new platform technology based on the binding of proteins/peptides to the outer surface of PEGylated liposomes (PEGLip) is then presented. Binding of proteins to PEGLip is non-covalent, highly specific and dependent on an amino acid consensus sequence within the proteins. Association of proteins with PEGLip results in substantial enhancement of the pharmacodynamic properties of proteins following administration. This has been demonstrated in preclinical studies and clinical trials with coagulation factors VIII and VIIa. It has also been demonstrated in preclinical studies with granulocyte colony-stimulating factor. A mechanism is presented that explains the improvements in hemostatic efficacy of PEGLip-formulated coagulation factors VIII and VIIa. WHAT THE READER WILL GAIN: The reader will gain an understanding of the advantages and disadvantages of each of the approaches discussed. TAKE HOME MESSAGE: PEGLip formulation is an important new approach to improve the pharmacodynamics of protein drugs. This approach may be applied to further therapeutic proteins in the future.
Asunto(s)
Química Farmacéutica/métodos , Factor VIII/administración & dosificación , Factor VIIa/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Liposomas/síntesis química , Proteínas/administración & dosificación , Animales , Anticoagulantes/administración & dosificación , Evaluación de Medicamentos , Evaluación Preclínica de Medicamentos , Humanos , Liposomas/administración & dosificación , Modelos Biológicos , Modelos Moleculares , Polietilenglicoles/administración & dosificación , Polietilenglicoles/química , Proteínas/químicaRESUMEN
We have previously shown that formulation of coagulation factor VIII and activated factor VII with PEGylated liposomes (PEGLip) results in an extension of circulation time and an increase in hemostatic efficacy. Here we identified additional proteins that associate with PEGLip, including granulocyte colony-stimulating factor (G-CSF). Surface plasmon resonance analyses indicated that G-CSF bound noncovalently but with high affinity and specificity to PEGLip. A pharmacokinetic study in mice demonstrated that PEGLip formulation of G-CSF extended its circulation time and resulted in higher G-CSF levels several hours after both subcutaneous and intravenous injection. PEGLip-formulated G-CSF had a significantly improved efficacy in the mobilization of hematopoietic stem cells (HSC) from the bone marrow to the peripheral blood. The results suggest that PEGLip-formulated G-CSF may function as an effective and safe tool for the mobilization of HSC prior to bone marrow transplantation. We also identified an amino acid sequence present in proteins that associate with PEGLip but absent from those that do not. A peptide based on this consensus sequence bound PEGLip. The results suggest that PEGLip formulation may serve as a platform for the delivery of additional short-half-life proteins/peptides having the relevant consensus sequence.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/farmacología , Movilización de Célula Madre Hematopoyética/métodos , Células Madre Hematopoyéticas/efectos de los fármacos , Polietilenglicoles/química , Secuencia de Aminoácidos , Animales , Sitios de Unión , Secuencia de Consenso , Citometría de Flujo , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/sangre , Factor Estimulante de Colonias de Granulocitos/química , Humanos , Inyecciones Intravenosas , Liposomas , Masculino , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Unión Proteica , Proteínas RecombinantesRESUMEN
Haemophilia A is a bleeding disorder caused by the lack of factor VIII (FVIII). We report the prolongation of exogenous FVIII circulation time and haemostatic efficacy by its formulation with PEGylated liposomes (PEGLip). FVIII binds non-covalently but with high affinity in a specific mode with the external surface of PEGLip neither losing its activity nor its binding to von Willebrand Factor. Experiments in haemophilic and non-haemophilic mice indicate that the circulation time and clotting efficacy of PEGLip-formulated exogenous FVIII (PEGLip-FVIII) are significantly enhanced over those of free FVIII. The data support the feasibility of using PEGLip-FVIII to extend the duration of haemostatic efficacy in the treatment of haemophilia A.