RESUMEN
Cystic fibrosis (CF) is a serious genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Approved small molecule therapies benefit the majority of people with CF (pwCF), but unfortunately not all. Gene addition offers a mutation agnostic treatment option for all pwCF. SP-101 is an adeno-associated virus gene therapy vector (AAV2.5T) that has been optimized for efficient human airway cell transduction, and that contains a functional and regulated shortened human CFTR minigene (hCFTRΔR) with a small synthetic promoter/enhancer. To understand SP-101 airway distribution, activity, and the associated immune response, in vivo studies were performed in wild-type and CF ferrets. After single dose inhaled delivery of SP-101, followed by single dose inhaled doxorubicin (an AAV transduction augmenter) or saline, SP-101 vector genomes were detected throughout the respiratory tract. hCFTRΔR mRNA expression was highest in ferrets also receiving doxorubicin and persisted for the duration of the study (13 weeks). Pre-existing mucus in the CF ferrets did not present a barrier to effective transduction. Binding and neutralizing antibodies to the AAV2.5T capsid were observed regardless of doxorubicin exposure. Only a portion of ferrets exhibited a weak T-cell response to AAV2.5T and no T-cell response was seen against hCFTRΔR. These data strongly support the continued development of inhaled SP-101, followed by inhaled doxorubicin, for the treatment of CF.
Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística , Fibrosis Quística , Dependovirus , Doxorrubicina , Hurones , Terapia Genética , Vectores Genéticos , Transgenes , Animales , Fibrosis Quística/terapia , Fibrosis Quística/genética , Doxorrubicina/farmacología , Doxorrubicina/administración & dosificación , Doxorrubicina/uso terapéutico , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Dependovirus/genética , Administración por Inhalación , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Humanos , Modelos Animales de Enfermedad , Expresión GénicaRESUMEN
CD33 is expressed in 90% of patients with acute myeloid leukemia (AML), and its extracellular portion consists of a V domain and a C2 domain. A recent study showed that a single nucleotide polymorphism (SNP), rs12459419 (C > T), results in the reduced expression of V domain-containing CD33 and limited efficacy of V domain-binding anti-CD33 antibodies. We developed JNJ-67571244, a novel human bispecific antibody capable of binding to the C2 domain of CD33 and to CD3, to induce T-cell recruitment and CD33+ tumor cell cytotoxicity independently of their SNP genotype status. JNJ-67571244 specifically binds to CD33-expressing target cells and induces cytotoxicity of CD33+ AML cell lines in vitro along with T-cell activation and cytokine release. JNJ-67571244 also exhibited statistically significant antitumor activity in vivo in established disseminated and subcutaneous mouse models of human AML. Furthermore, this antibody depletes CD33+ blasts in AML patient blood samples with concurrent T-cell activation. JNJ-67571244 also cross-reacts with cynomolgus monkey CD33 and CD3, and dosing of JNJ-67571244 in cynomolgus monkeys resulted in T-cell activation, transient cytokine release, and sustained reduction in CD33+ leukocyte populations. JNJ-67571244 was well tolerated in cynomolgus monkeys up to 30 mg/kg. Lastly, JNJ-67571244 mediated efficient cytotoxicity of cell lines and primary samples regardless of their SNP genotype status, suggesting a potential therapeutic benefit over other V-binding antibodies. JNJ-67571244 is currently in phase 1 clinical trials in patients with relapsed/refractory AML and high-risk myelodysplastic syndrome.
Asunto(s)
Leucemia Mieloide Aguda , Linfocitos T , Animales , Dominios C2 , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Macaca fascicularis , Lectina 3 Similar a Ig de Unión al Ácido Siálico/genética , Linfocitos T/metabolismoRESUMEN
The recent Scientific Committee on Health, Environmental and Emerging Risks Final Opinion on "The need for nonhuman primates in biomedical research, production and testing of products and devices" (2017 SCHEER) highlights approaches that could significantly contribute to the replacement, reduction, and refinement of nonhuman primate (NHP) studies. Initiatives that have the potential to affect NHP welfare and/or their use are expected to be appropriate, fair, and objective and publicly disseminated information focused on NHPs in biomedical research, which includes toxicologic and pathologic research and testing, should be objectively evaluated by stakeholder scientists, researchers, and veterinarians. Thus, IQ Consortium member companies convened to develop an informed and objective response, focusing on identifying areas of agreement, potential gaps, or missing information in 2017 SCHEER. Overall, the authors agree that many positions in the 2017 SCHEER Opinion generally align with industry views on the use of NHPs in research and testing, including the ongoing need of NHPs in many areas of research. From the perspective of the IQ Consortium, there are several topics in the 2017 SCHEER that merit additional comment, attention, or research, as well as consideration in future opinions.
Asunto(s)
Alternativas al Uso de Animales/tendencias , Investigación Biomédica/métodos , Evaluación Preclínica de Medicamentos/tendencias , Primates , Alternativas al Uso de Animales/ética , Alternativas al Uso de Animales/legislación & jurisprudencia , Bienestar del Animal , Animales , Bioética , Investigación Biomédica/ética , Investigación Biomédica/legislación & jurisprudencia , Evaluación Preclínica de Medicamentos/ética , Evaluación Preclínica de Medicamentos/métodos , Unión Europea , Regulación GubernamentalRESUMEN
An Innovation and Quality (IQ) Consortium focus group conducted a cross-company survey to evaluate current practices and perceptions around the use of animal models of disease (AMDs) in nonclinical safety assessment of molecules in clinical development. The IQ Consortium group is an organization of pharmaceutical and biotechnology companies with the mission of advancing science and technology. The survey queried the utilization of AMDs during drug discovery in which drug candidates are evaluated in efficacy models and limited short-duration non-Good Laboratory Practices (GLP) toxicology testing and during drug development in which drug candidates are evaluated in GLP toxicology studies. The survey determined that the majority of companies used AMDs during drug discovery primarily as a means for proactively assessing potential nonclinical safety issues prior to the conduct of toxicology studies, followed closely by the use of AMDs to better understand toxicities associated with exaggerated pharmacology in traditional toxicology models or to derisk issues when the target is only expressed in the disease state. In contrast, the survey results indicated that the use of AMDs in development is infrequent, being used primarily to investigate nonclinical safety issues associated with targets expressed only in disease states and/or in response to requests from global regulatory authorities.
Asunto(s)
Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos/métodos , Industria Farmacéutica , Animales , Toma de Decisiones en la Organización , Evaluación Preclínica de Medicamentos/estadística & datos numéricos , Industria Farmacéutica/legislación & jurisprudencia , Industria Farmacéutica/organización & administración , Industria Farmacéutica/normas , Regulación Gubernamental , Encuestas y CuestionariosRESUMEN
alpha(1)-Adrenoceptor function and density in isolated thoracic aorta were measured during the course of streptozotocin-induced diabetes in rats. Diabetes was induced by a single tail vein injection of streptozotocin (60 mg/kg) and was verified by four measures (blood glucose level, increase in food intake, increase in water intake, and characteristic weight changes). Diabetes produced a significant increase in isolated aorta sensitivity to alpha(1)-adrenoceptor activation, manifested as a significant (p < 0.05) decrease in the A(50) value for phenylephrine and an increase in the A(50) (control)/A(50) (diabetic) ratio (1.2, 1.7, and 2.0, respectively) with increasing length of diabetes (4, 12 and 52 weeks). There was a large and biphasic change in receptor density (B(max)), without a significant change (p > 0.05) in either agonist (phenylephrine) or antagonist (prazosin) affinities (K(A) and pA(2) values, respectively). These results suggest compensatory mechanisms in receptor number and abnormalities in 2nd messenger transduction and can help direct efforts for improving antihypertensive or other pharmacological therapy for diabetic patients.
Asunto(s)
Aorta Torácica/fisiopatología , Diabetes Mellitus Experimental/metabolismo , Receptores Adrenérgicos alfa 1/biosíntesis , Agonistas de Receptores Adrenérgicos alfa 1 , Antagonistas de Receptores Adrenérgicos alfa 1 , Animales , Sitios de Unión , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/fisiopatología , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Contracción Isométrica/efectos de los fármacos , Masculino , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/fisiopatología , Fenilefrina/farmacología , Prazosina/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Estreptozocina , Factores de TiempoRESUMEN
The primary therapeutic goal for the treatment of diabetes is maintenance of a long-term, near-normoglycemic condition and prevention of the onset or progression of the complications associated with the disease. Although several analogs of human insulin have been developed, the currently prescribed long-acting insulin analogs do not provide a stable basal glycemia for more than a few hours. Here, we report the development of Albulin, a long-acting insulin analog obtained by direct gene fusion of a single-chain human insulin to human serum albumin. Albulin showed an elimination t(1/2) of approximately 7 h in normoglycemic mice. In vitro pharmacodynamic profiles for Albulin characterized by receptor binding, inhibition of gluconeogenesis, induction of glucose uptake, and global regulation of gene expression in relevant cell types showed that Albulin produced similar activity profiles compared with that of recombinant human insulin. A single Albulin administration in vivo normalized blood glucose level in diabetic mice in a relatively peakless and sustained (24-h) fashion. A further reduction in glucose levels was achieved by administering a recombinant human insulin a few hours after Albulin injection in mice, indicating the potential for Albulin therapy in combination with available fast-acting insulin derivatives. In summary, Albulin displays characteristics of a potent long-acting insulin analog that can be evaluated for use as a novel insulin therapy for patients with insulin-dependent diabetes.
Asunto(s)
Insulina/genética , Insulina/farmacocinética , Proteínas Recombinantes de Fusión/farmacocinética , Albúmina Sérica/genética , Albúmina Sérica/farmacocinética , Células 3T3 , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Animales , Clonación Molecular , Escherichia coli , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Genes Sintéticos , Glucosa/metabolismo , Humanos , Insulina/farmacología , Insulina de Acción Prolongada , Insulina Regular Humana , Masculino , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena de la Polimerasa , Receptor de Insulina/metabolismo , Proteínas Recombinantes de Fusión/farmacología , Albúmina Sérica/farmacología , Albúmina Sérica HumanaRESUMEN
Certain ribonucleases (RNases), such as eosinophil-derived neurotoxin, are associated with pathological conditions (e.g. asthma and inflammatory bowel disease) and can even be overtly cyto(neuro)toxic. It has been proposed that small-molecule inhibitors should have therapeutic utility. We used isothermal titration microcalorimetry to characterize reversible inhibitor cytidine 2'-monophosphate (2'-CMP) binding to RNase-A in a multi-ion buffer at 37 degrees as a representative system. The estimated parameters were: K(d)=13.9 microM; DeltaG degrees =-6.90 kcal/mol; DeltaH degrees =-15.7 kcal/mol; and DeltaS degrees =-0.028 kcal/mol-K ('enthalpy-driven' interaction). These data should assist drug design of small-molecule inhibitors of homologous RNase catalytic domains.
Asunto(s)
Citidina Monofosfato/farmacología , Inhibidores Enzimáticos/farmacología , Ribonucleasa Pancreática/antagonistas & inhibidores , Animales , Sitios de Unión , Tampones (Química) , Bovinos , Metales/metabolismo , Ribonucleasa Pancreática/metabolismo , TermodinámicaRESUMEN
Present antipsychotic drugs, whose clinical activity correlates with direct binding to dopamine D2 or other receptors, alleviate some of the symptoms of schizophrenia, but not all and not completely in many patients. In continuation of our overview of potential novel antipsychotic pharmacotherapy that would be based upon indirect modulation of dopamine or other neurotransmitter functioning, we focus in this article on the postulated use of retinoid analogs as novel antipsychotic agents. Several lines of evidence can be viewed as implicating retinoid dysregulation in schizophrenia, either as a causative or contributory factor. It has been proposed that using retinoid analogs to alter the downstream expression of dopamine D2 receptors might represent a novel approach to the treatment of the disease or amelioration of symptoms when used either as monotherapy or as adjunct pharmacotherapy to dopamine D2 receptor antagonists.
Asunto(s)
Antipsicóticos/farmacología , Receptores de Dopamina D2/efectos de los fármacos , Retinoides/farmacología , Esquizofrenia/tratamiento farmacológico , Antipsicóticos/química , Antipsicóticos/uso terapéutico , Quimioterapia Combinada , Humanos , Receptores de Dopamina D2/genética , Receptores de Dopamina D2/fisiología , Retinoides/química , Retinoides/uso terapéutico , Esquizofrenia/etiología , Esquizofrenia/metabolismo , Transcripción GenéticaRESUMEN
Ribonucleases (RNases) possess a variety of biological activities and, under certain conditions, are deleterious. Hence, design of selective inhibitors has been suggested as a strategy for treating RNase-related disorders. In the present study, isothermal titration calorimetry was used to measure ion effects on binding thermodynamics of the RNase A competitive inhibitor 2'-CMP as a representative system. The reaction cell (37 degrees C) contained dialyzed RNase A (0.04-0.05 mM) in buffered solution (pH 5.5) of 50 mM Na(+), K(+), Ca(2+), or Mg(2+) acetate, verified spectrophotometrically. Thirty-five sequential injections (4 microl each, 3 min apart) were made of 2'-CMP (1.2 mM) in ion-matching buffer. The data were corrected for heat of dilution. There was a 1:1 interaction in each case. The estimated parameters (+/-S.D.) were: K(d) = 4.84 +/- 0.29 microM (Na(+)); 5.62 +/- 0.98 microM (K(+)); 24.44 +/- 6.96 microM (Ca(2+)); 28.74 +/- 0.43 microM (Mg(2+)); DeltaG(o) = -7.541 +/- 0.037 kcal/mol (Na(+)); -7.458 +/- 1.03 kcal/mol (K(+)); -6.574 +/- 0.173 kcal/mol (Ca(2+)); -6.442 +/- 0.009 kcal/mol (Mg(2+)); DeltaH(o) = -22.357 +/- 1.189 kcal/mol (Na(+)); -21.917 +/- 0.891 kcal/mol (K(+)); -20.223 +/- 1.503 kcal/mol (Ca(2+)); -26.570 +/- 1.579 kcal/mol (Mg(2+)); and DeltaS(o) = -0.048 +/- 0.004 kcal/mol-K (Na(+)); -0.047 +/- 0.003 kcal/mol-K (K(+)); -0.044 +/- 0.005 kcal/mol-K (Ca(2+)); -0.065 +/- 0.005 kcal/mol-K (Mg(2+)). Thus, all reactions were enthalpy-driven. Despite a 5-fold difference in K(d) between mono- and divalent ions, the ratio of ion hydration DeltaG(o) to K(d) was constant. These data should be useful for molecular modeling and suggest that inhibitor activity will be a function of cellular conditions (normal or pathological).