RESUMO
Background: Autotaxin-lysophosphatidic acid (ATX-LPA) signaling has a predominant role in immunological and fibrotic processes, including cancer. Several ATX inhibitors and LPA receptor antagonists have been clinically evaluated, but none in patients with solid tumors. Many cancers are burdened with a high degree of fibrosis and an immune desert phenotype (so-called 'cold' tumors). In these cold tumors, the fibrotic stroma provides an intrinsic cancer-supporting mechanism. Furthermore, the stroma prevents penetration and limits the effectiveness of existing therapies. IOA-289 is a novel ATX inhibitor with a unique chemical structure, excellent potency and an attractive safety profile. Materials and methods: In vitro and in vivo pharmacology studies have been carried out to elucidate the pharmaceutical properties and mechanism of action of IOA-289. A phase I clinical study in healthy volunteers was carried out to determine the pharmacokinetics and pharmacodynamics of IOA-289 following a single oral dose. Results: In vitro and in vivo studies showed that IOA-289 is a potent inhibitor of ATX and, as a monotherapy, is able to slow progression of lung fibrosis and tumor growth in mouse models. In a clinical study, IOA-289 showed a dose-dependent increase in plasma exposure levels and a corresponding decrease in circulating LPA. Conclusions: Our data show that IOA-289 is a novel ATX inhibitor with a unique chemical structure, excellent potency and an attractive safety profile. Our data support the further development of IOA-289 as a novel therapeutic approach for the treatment of cancer, particularly those with a high fibrotic and immunologically cold phenotype.
RESUMO
IOA-289 is a novel small molecule inhibitor of autotaxin developed as a first-in-class therapy of fibrotic pathologies including cancer. A method for quantitation of IOA-289 in human plasma was developed using a stable isotope labeled compound ([13C4]IOA-289) as internal standard. The analytes were extracted from human plasma by protein precipitation and the analysis was performed by liquid chromatography coupled with tandem mass spectrometric detection (LCMS/MS). The chromatographic separation was performed with a gradient elution from a BEH C18 column and under these conditions the retention time and the run time were 1 and 2â¯min, respectively. The assay was fully validated over the range 3-3000â¯ng/mL, proved to be accurate, precise and selective and was successfully applied to quantitate IOA-289 in plasma samples from subjects in a first-in-humanclinical trial.
Assuntos
Plasma , Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida/métodos , Humanos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodosRESUMO
The heme group of myeloperoxidase is covalently linked via two ester bonds to the protein and a unique sulfonium ion linkage involving Met(243). Mutation of Met(243) into Thr, Gln, and Val, which are the corresponding residues of eosinophil peroxidase, lactoperoxidase, and thyroid peroxidase, respectively, and into Cys was performed. The Soret band in the optical absorbance spectrum in the oxidized mutants is now found at approximately 411 nm. Both the pyridine hemochrome spectra and resonance Raman spectra of the mutants are affected by the mutation. In the Met(243) mutants the affinity for chloride has decreased 100-fold. All mutants have lost their chlorination activity, except for the M243T mutant, which still has 15% activity left. By Fourier transform infared difference spectroscopy it was possible to specifically detect the (13)CD(3)-labeled methionyl sulfonium ion linkage. We conclude that the sulfonium ion linkage serves two roles. First, it serves as an electron-withdrawing substituent via its positive charge, and, second, together with its neighboring residue Glu(242), it appears to be responsible for the lower symmetry of the heme group and distortion from the planar conformation normally seen in heme-containing proteins.
Assuntos
Peroxidase/metabolismo , Compostos de Sulfônio/metabolismo , Substituição de Aminoácidos , Animais , Células CHO , Cloretos/metabolismo , Cricetinae , Espectroscopia de Ressonância de Spin Eletrônica , Heme/metabolismo , Hidrocarbonetos , Concentração de Íons de Hidrogênio , Metano/análogos & derivados , Metano/metabolismo , Modelos Químicos , Mutagênese Sítio-Dirigida , Oxirredução , Peroxidase/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman , TransfecçãoRESUMO
OBJECTIVE: To compare the attitudes of couples whose children were conceived through artificial insemination by donor (AID) in 1980 and in 1996. DESIGN: Replication study. SETTING: Infertility clinic of the Utrecht university hospital. PATIENT(S): Couples who conceived a child through AID in 1980 (n = 134) or in 1996 (n = 110). INTERVENTION(S): Anonymous questionnaires. MAIN OUTCOME MEASURE(S): Answers to questionnaires. RESULT(S): Couples who conceive a child through AID still strongly prefer (84%) absolute anonymity of the donor. In both 1980 and 1996, most couples (approximately 80%) decided not to inform their child about the nature of his or her conception. In 1996, couples who considered informing their child hesitated significantly less and showed significantly more openness toward others. In addition, more couples wanted unidentifiable data about the donor and considered it more important to use the same donor for a subsequent child. CONCLUSION(S): Between 1980 and 1996, the number of couples who conceived a child through AID and adhered to absolute anonymity of the donor and secrecy toward the child remained the same, whereas their openness toward others and desire for unidentifiable data about the donor increased.
Assuntos
Atitude , Fertilização/fisiologia , Inseminação Artificial Heteróloga , Confidencialidade , Feminino , Humanos , Masculino , Estudos Retrospectivos , Inquéritos e QuestionáriosRESUMO
A sol-particle immunoassay (SPIA) was developed for determination of anti-rubella antibodies. The test is based on antibody-coated gold sol particles, which may agglutinate in the presence of rubella haemagglutinin antigen. The agglutination which gives a decrease in optical density can be inhibited by specific antibodies present in sera incubated with the immunochemical reagents. The test is performed in microtitration plates and results can be read in a plate-reader. Clinical validation studies were performed comparing the anti-rubella SPIA with the haemagglutination inhibition test and with the latex agglutination test. The sensitivity of the SPIA was calculated to be 99%; the specificity of the test was at least 99.5%. The anti-rubella SPIA is highly suitable for large-scale screening to determine the rubella immune-status.