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Enzymatic Stability (enzymatic + stability)
Selected AbstractsSynthesis, CD Spectra, and Enzymatic Stability of ,2 -Oligoazapeptides Prepared from (S)-2-Hydrazino Carboxylic Acids Carrying the Side Chains of Val, Ala, and LeuHELVETICA CHIMICA ACTA, Issue 12 2003Gérald Lelais , -Peptides offer the unique possibility to incorporate additional heteroatoms into the peptidic backbone (Figs.,1 and 2). We report here the synthesis and spectroscopic investigations of ,2 -peptide analogs consisting of (S)-3-aza- , -amino acids carrying the side chains of Val, Ala, and Leu. The hydrazino carboxylic acids were prepared by a known method: Boc amidation of the corresponding N -benzyl- L - , -amino acids with an oxaziridine (Scheme,1). Couplings and fragment coupling of the 3-benzylaza- ,2 -amino acids and a corresponding tripeptide (N -Boc/C -OMe strategy) with common peptide-coupling reagents in solution led to ,2 -di, ,2 -tri-, and ,2 -hexaazapeptide derivatives, which could be N -debenzylated (4,9; Schemes,2,4). The new compounds were identified by optical rotation, and IR, 1H- and 13C-NMR, and CD spectroscopy (Figs.,4 and 5) and high-resolution mass spectrometry, and, in one case, by X-ray crystallography (Fig.,3). In spite of extensive measurements under various conditions (temperatures, solvents), it was not possible to determine the secondary structure of the ,2 -azapeptides by NMR spectroscopy (overlapping and broad signals, fast exchange between the two types of NH protons!). The CD spectra of the N -Boc and C -OMe terminally protected hexapeptide analog 9 in MeOH and in H2O (at different pH) might arise from a (P)- 314 -helical structure. The N -Boc- ,2 -tri and N -Boc- ,2 -hexaazapeptide esters, 7 and 9, were shown to be stable for 48,h against the following peptidases: pronase, proteinase,K, chymotrypsin, trypsin, carboxypeptidase,A, and 20S proteasome. [source] C5-Functionalized LNA: Unparalleled Hybridization Properties and Enzymatic StabilityCHEMBIOCHEM, Issue 17 2009Michael E. Østergaard The best of both worlds: C5-functionalized locked nucleic acids (LNAs) synergistically combine the beneficial features of C5-alkynyl DNAs and conventional antisense LNA,they are straightforward to make and yet exhibit unparalleled thermal affinity toward RNA targets, extraordinary discrimination of singly mismatched RNA strands, and superb stability against degradation by 3,-exonucleases. [source] Enzymatic stability of 2,-ethylcarbonate-linked paclitaxel in serum and conversion to paclitaxel by rabbit liver carboxylesterase for use in prodrug/enzyme therapyBIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 5 2008Tadatoshi Tanino Abstract In prodrug/enzyme therapy for cancer, information on the sensitivity of hydrolytic enzymes to prodrug is required to reduce adverse effects of the parental drug and to find the activating enzyme. The aim of this study was to characterize the enzymatic stability of 2,-ethylcarbonate-linked paclitaxel (TAX-2,-Et) in the sera of several different species including humans. TAX-2,-Et disposition in serum was kinetically analysed using models with hydrolytic and/or degradation processes. To further evaluate the capability of liver carboxylesterases (CESs) in TAX-2,-Et hydrolysis, a CES isolated from rabbit liver (Ra-CES) was utilized as a model enzyme. Rat serum provided rapid enzymatic hydrolysis of TAX-2,-Et with a half-life of 4 min. The degradation of paclitaxel (TAX) (degradation rate constant, 0.16,h,1) was accompanied by the formation of an unknown compound. The conversion to TAX was almost completely inhibited by phenylmethyl sulfonylfluoride (PMSF) and bis(p-nitrophenyl) phosphate (BNPP). In human and rabbit sera, the degradation rate constant of TAX-2,-Et was 5.1,×,10,2 and 0.15,h,1, respectively, when excepting hydrolysis. The degradation products had the same molecular weight as TAX-2,-Et. The amount of TAX produced accounted for only 8,11% of the decrease in TAX-2,-Et after a 9 h exposure to rabbit or human serum. PMSF, but not BNPP, inhibited more than 90% of the TAX production in a 1.5,h incubation with human or rabbit serum. Ra-CES enzyme converted TAX-2,-Et to TAX with Vmax and Km of 74.7±13.8 nmol/min/mg protein and 8.8±2.8 µM, respectively. These results indicate that TAX-2,-Et is sensitive to serum CESs, but not cholinesterases. However, serum CESs show species-dependent hydrolysis of TAX-2,-Et. Although human serum allows the slow release of TAX, TAX-2,-Et is expected to reduce the side-effects of TAX. The Ra-CES enzyme is capable of hydrolysing TAX-2,-Et, which may be beneficial for the development of a TAX-2,-Et/enzyme therapy strategy for ovarian cancer. Copyright © 2008 John Wiley & Sons, Ltd. [source] Synthesis and Stability in Biological Media of 1H -Imidazole-1-carboxylates of ROS203, an Antagonist of the Histamine H3 ReceptorCHEMISTRY & BIODIVERSITY, Issue 1 2008Mirko Rivara Abstract A series of carbamate derivatives of the H3 antagonist ROS203 (1) were prepared, and their lipophilicity and steric hindrance were modulated by introducing linear or branched alkyl chains of various lengths. In vitro stability studies were conducted to evaluate how structural modulations affect the intrinsic reactivity of the carbamoyl moiety and its recognition by metabolic enzymes. Linear alkyl carbamates were the most susceptible to enzymatic hydrolysis, with bioconversion rates being higher in rat liver and plasma. Chain ramification significantly enhanced the enzymatic stability of the set, with two derivatives (1g and 1h) being more stable by a factor of 8,40 than the ethyl carbamate 1a. Incubation with bovine serum albumin (BSA) showed a protective role of proteins on chemical and porcine-liver esterase (PLE)-catalyzed hydrolysis. Ex vivo binding data after i.v. administration of 1h revealed prolonged displacement of the labeled ligand [3H]-(R)- , -methylhistamine ([3H]RAMHA) from rat-brain cortical membranes, when compared to 1. However, the high rates of bioconversion in liver, as well as the chemical instability of 1h, suggest that further work is needed to optimize the enzymatic and chemical stability of these compounds. [source] Lipophilicity Plays a Major Role in Modulating the Inhibition of Monoamine Oxidase B by 7-Substituted CoumarinsCHEMISTRY & BIODIVERSITY, Issue 2 2006Angelo Carotti Abstract A series of coumarin derivatives (1,22), bearing at the 7-position ether, ketone, ester, carbamate, or amide functions of varying size and lipophilicity, were synthesized and investigated for their in vitro monoamine oxidase-A and -B (MAO-A and -B) inhibitory activities. Most of the compounds acted preferentially as MAO-B inhibitors, with IC50 values in the micromolar to low-nanomolar range. A structure,activity-relationship (SAR) study highlighted lipophilicity as an important property modulating the MAO-B inhibition potency of 7-substituted coumarins, as shown by a linear correlation (n=20, r2=0.72) between pIC50 and calculated log P values. The stability of ester-containing coumarin derivatives in rat plasma provided information on factors that either favor (lipophilicity) or decrease (steric hindrance) esterase-catalyzed hydrolysis. Two compounds (14 and 22) were selected to investigate how lipophilicity and enzymatic stability may affect in vivo MAO activities, as assayed ex vivo in rat. The most-potent and -selective MAO-B inhibitor 22 (=7-[(3,4-difluorobenzyl)oxy]-3,4-dimethyl-1-benzopyran-2(2H)-one) within the examined series significantly inhibited (>60%) ex vivo rat-liver and striatal MAO-B activities 1,h after intraperitoneal administration of high doses (100 and 300,,mol kg,1), revealing its ability to cross the blood,brain barrier. At the same doses, liver and striatum MAO-A was less inhibited in vivo, somehow reflecting MAO-B selectivity, as assessed in vitro. In contrast, the metabolically less stable derivative 14, bearing an isopropyl ester in the lateral chain, had a weak effect on hepatic MAO-B activity in vivo, and none on striatal MAO-B, but, surprisingly, displayed inhibitory effects on MAO-A in both peripheral and brain tissues. [source] | ||||||||||