ABSTRACT
Pharmacogenomic tests used to guide clinical treatment for major depressive disorder (MDD) must be thoroughly validated. One important assessment of validity is the ability to predict medication blood levels, which reflect altered metabolism. Historically, the metabolic impact of individual genes has been evaluated; however, we now know that multiple genes are often involved in medication metabolism. Here, we evaluated the ability of individual pharmacokinetic genes (CYP2C19, CYP2D6, CYP3A4) and a combinatorial pharmacogenomic test (GeneSight Psychotropic®; weighted assessment of all three genes) to predict citalopram/escitalopram blood levels in patients with MDD. Patients from the Genomics Used to Improve DEpression Decisions (GUIDED) trial who were taking citalopram/escitalopram at screening and had available blood level data were included (N=191). In multivariate analysis of the individual genes and combinatorial pharmacogenomic test separately (adjusted for age, smoking status), the F statistic for the combinatorial pharmacogenomic test was 1.7 to 2.9-times higher than the individual genes, showing that it explained more variance in citalopram/escitalopram blood levels. In multivariate analysis of the individual genes and combinatorial pharmacogenomic test together, only the combinatorial pharmacogenomic test remained significant. Overall, this demonstrates that the combinatorial pharmacogenomic test was a superior predictor of citalopram/escitalopram blood levels compared to individual genes.
Subject(s)
Antidepressive Agents/blood , Antidepressive Agents/pharmacokinetics , Citalopram/blood , Citalopram/pharmacokinetics , Cytochrome P-450 CYP2C19/genetics , Cytochrome P-450 CYP2D6/genetics , Cytochrome P-450 CYP3A/genetics , Depressive Disorder, Major/drug therapy , Selective Serotonin Reuptake Inhibitors/blood , Selective Serotonin Reuptake Inhibitors/pharmacokinetics , Adult , Algorithms , Antidepressive Agents/therapeutic use , Citalopram/therapeutic use , Cytochrome P-450 CYP2D6/metabolism , Cytochrome P-450 CYP3A/metabolism , Depressive Disorder, Major/blood , Depressive Disorder, Major/genetics , Female , Humans , Male , Middle Aged , Pharmacogenetics , Pharmacogenomic Testing , Selective Serotonin Reuptake Inhibitors/therapeutic use , Treatment OutcomeABSTRACT
Modification of nuclear and cytoplasmic proteins by the addition or removal of O-GlcNAc dynamically impacts multiple biological processes. Here, we present the development of a chemoenzymatic histology method for the detection of O-GlcNAc in tissue specimens. We applied this method to screen murine organs, uncovering specific O-GlcNAc distribution patterns in different tissue structures. We then utilized our histology method for O-GlcNAc detection in human brain specimens from healthy donors and donors with Alzheimer's disease and found higher levels of O-GlcNAc in specimens from healthy donors. We also performed an analysis using a multiple cancer tissue array, uncovering different O-GlcNAc levels between healthy and cancerous tissues, as well as different O-GlcNAc cellular distributions within certain tissue specimens. This chemoenzymatic histology method therefore holds great potential for revealing the biology of O-GlcNAc in physiopathological processes.
Subject(s)
Acetylglucosamine/analysis , Histological Techniques , Organ Specificity , Acetylglucosamine/metabolism , Alzheimer Disease/metabolism , Animals , Brain/metabolism , Brain Chemistry , Humans , Mice , Neoplasms/chemistry , Neoplasms/metabolism , Tissue DistributionABSTRACT
A panel of 1-deoxynojirimycin (DNJ) N-linked peptides were synthesized. Their IC50 values were measured in vitro against α-glucosidases I and II and were found to be in the micromolar range for both isozymes, and better than that of the iminosugar NB-DNJ (miglustat, 3) against α-glucosidase II. Cell-based studies revealed that although the free iminosugar 3 is most effective at disrupting N-linked glycan processing for short-term incubations (one day), when the cell-based studies were extended to three days, the DNJ N-linked tetrapeptide KDEL, which is an endoplasmic reticulum (ER)-retaining sequence, performed far better than 3. In low inhibitor washout studies, NB-DNJ inhibition was decreased to zero after 24 h, but DNJ-KDEL retained 13 % activity. This method offers a general approach for targeting drugs to the ER and prolonging their activity. Moreover, it is modular, so as new iminosugars of increased potency are discovered, they can be added to this template for targeting.
Subject(s)
Enzyme Inhibitors/chemical synthesis , Glucosamine/analogs & derivatives , Peptides/chemistry , Polysaccharides/metabolism , Amino Acid Sequence , Animals , CHO Cells , Cricetinae , Cricetulus , Endoplasmic Reticulum/enzymology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/metabolism , Glucosamine/chemistry , Glycosylation , Imino Sugars/chemistry , Nitrogen/chemistry , Polysaccharides/chemistry , Protein Binding , alpha-Glucosidases/chemistry , alpha-Glucosidases/metabolismABSTRACT
Attachment of carbohydrates to oligonucleotides has proven to induce receptor-mediated endocytosis. A facile method for the formation of covalent linkages between glycans and oligonucleotides is herein described. Thus, use of 3,4-diethoxy-3-cyclobutene-1,2-dione as a linking reagent provides easy conjugation between carbohydrates bearing an amino group at the reducing end and oligonucleotides bearing an aminoalkyl modification.
Subject(s)
Carbohydrates/chemical synthesis , Cyclobutanes/chemical synthesis , Glycoconjugates/chemical synthesis , Oligonucleotides/chemical synthesis , Cross-Linking Reagents/chemistry , Drug Delivery Systems , Glycosylation , Oligonucleotides/metabolism , Oligonucleotides/therapeutic use , Polysaccharides/metabolismABSTRACT
Advanced cleaning formulations, such as liquid laundry packages, consist of many components that require a complex mixture of interfacial actives including silicones for foam control, bleach for brightening, and enzymes for stain removal. Many of these ingredients are mutually incompatible, particularly in liquid formulations where they can be in intimate contact over extended periods of time. Solid dispersions of a prototypical bleach, NaBO3, in silicone polyether surfactants were shown to be very stable over time, even in the presence of water-in-silicone (D(4)) emulsions containing the enzyme alpha-chymotrypsin. Normally, perborates undergo rapid decomposition on contact with water. The rate of denaturation of the enzyme in the emulsion was similarly unaffected by the presence of the bleach until the emulsion was broken, unlike the case where the polyether surfactant was not present. The polyether surfactant thus protects the perborate from hydration and the enzyme from denaturing on contact with silicone oil until excess water and high shear are applied to the emulsion; protective mechanisms are discussed.
Subject(s)
Borates/chemistry , Chymotrypsin/chemistry , Silicone Oils/chemistry , Surface-Active Agents/chemistry , EmulsionsABSTRACT
We herein report a convenient synthesis of 3',5'-cyclic diguanylic acid via the modified H-phosphonate approach. The 1-(4-chlorophenyl)-4-ethoxypiperidin-4-yl (Cpep) group was used as protecting group for the 2'-hydroxy functions of ribonucleosides. Complete unblocking of the fully protected 3',5'-cyclic diguanylic acid gave cdiGMP as a homogeneous compound in an excellent yield.
