Drug Enantiomers Research Articles

Non-enzymatic dynamic kinetic resolution of racemic α-arylalkanoic acids: an advanced asymmetric synthesis of chiral nonsteroidal anti-inflammatory drugs (NSAIDs)

An efficient protocol was developed to produce chiral 2-arylalkanoic esters in high yields (up to 99%) from racemic carboxylic acids utilizing the racemization of the mixed-anhydrides generated from acid components with pivalic anhydride in the presence of an acyl-transfer catalyst. The present DKR involves the enantio-discriminating esterification of the racemic 2-arylalkanoic acids and the rapid racemization of the chiral 2-arylalkanoic acids under suitable reaction conditions using pivalic anhydride, diisopropylethylamine, and benzotetramisole (BTM) in a polar solvent, and this method was successfully applied for the preparation of pharmacodynamically active (S)-enantiomers of nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and naproxen.

Spectrophotometric Methods for Simultaneous Estimation of Thiocolchicoside and Dexketoprofen Trometamol in Pharmaceutical Dosage Form

INTRODUCTION Dexketoprofen trometamol (DKP) chemically, 2-amino-2-(hydroxymethyl) propane-1,3-diol; 2-(3-benzoylphenyl propanoic acid is a water-soluble salt of the dextrorotatory enantiomer or (S)-(+)-enantiomer of the nonsteroidal anti-inflammatory drug (NSAID) ketoprofen1,2. The enantiomer is a relatively new oral NSAID with analgesic, anti-inflammatory and anti-pyretic properties and is one of the most potent in vitro inhibitors of prostaglandin synthesis.

Combination of capillary electrophoresis, molecular modelling and nuclear magnetic resonance to study the interaction mechanisms between single-isomer anionic cyclodextrin derivatives and basic drug enantiomers in a methanolic background electrolyte

In order to improve our knowledge of the mechanisms of enantiomer recognition pattern in nonaqueous systems, an approach combining nonaqueous CE (NACE), molecular modelling and NMR was undertaken. Bupivacaine and propranolol were selected as model compounds and their interactions with two single-isomer highly charged β-CD derivatives, namely heptakis(2,3-di-O-methyl-6-O-sulfo)-β-CD (HDMS-β-CD) and heptakis(2,3-di-O-acetyl-6-O-sulfo)-β-CD (HDAS-β-CD), were studied. The CD-bupivacaine complexes were evaluated by 2-D Rotating-frame Overhauser Effect SpectroscopY (ROESY) experiments. From these experiments, it can be assumed that inclusion complexes are not formed, whatever the CD derivative used. Molecular modelling was performed at the RHF/MINI-1 or B3LYP/6-31G(d) level. External as well as inclusion type complexes with the alkyl chain of propranolol into both CD cavities were located. Interaction energies calculated for bupivacaine and propranolol correlated with the enantiomer migration order observed in the NACE experiments using both anionic CD derivatives. The interaction of propranolol with HDMS-β-CD or HDAS-β-CD gives rise to a family of external and inclusion complexes in which some are more probably obtained.

Development and validation of a reversed‐phase HPLC method for determination of lesatropane and enantiomeric impurity

Lesatropane is a novel muscarinic receptor agonist and is currently being under preclinical development in China as a single enantiomer drug for the treatment of primary glaucoma. A reversed-phase chiral HPLC method for determination of lesatropane and enantiomeric impurity was developed. Enantiomeric separation of lesatropane from its enantiomer (desatropane) was achieved in normal-phase mode with Chiralpak AD-H and in reversed-phase mode with Chiralpak AS-RH. The conditions using a Chiralpak AS-RH column and mobile phase of K(2) HPO(4) -KH(2) PO(4) (pH 7.0; 0.02 M)-acetonitrile (69:31, v/v) at a flow rate of 0.5 ml/min have been fully validated with satisfactory specificity, linearity, accuracy, and precision. The method was found to be suitable for the simultaneous quantitation of lesatropane and enantiomeric impurity desatropane.

Modeling multiple chemical equilibrium for reactive extraction of naproxen enantiomers with HP-β-CD as hydrophilic selector

Reactive extraction is an emerging technology for large-scale continuous resolution of drug enantiomers. The enantioselective extraction of R,S-naproxen by hydrophilic HP-β-CD in 1,2-dichloroethane was studied at 5 °C. The experimental data were described by a reactive extraction model with a homogeneous aqueous phase reaction of R,S-naproxen with HP-β-CD which couples a complete description of chemical equilibria in aqueous phase with the overall phase equilibria of the system. Important parameters of this model were determined experimentally. The physical distribution coefficients for molecular and ionic NAP were 0.041 and 1.730, respectively. Here we show that the efficiency of extraction depends strongly on two process variables including pH and HP-β-CD concentration. The model predictions are compared graphically with the results of previous experiments and there is a good agreement between each other. By the use of modeling and experiment, an optimized extraction condition with pH of 2.5 and HP-β-CD concentration of 0.1 mol/L was obtained with high enantioselectivity (α?) of 1.59 and performance factor (pf) of 0.049. The model gives a good means of predicting enantiomers partitioning over a range of experimental conditions.

The significance of differences in the potency of enantiomers of anti-acetylcholine drugs.

Abstract The anti-acetylcholine potencies of the dimethylaminoethyl and N/-methyl piperidin-4-yl esters of R and S 2-cyclohexyl-2-hydroxy-2-phenylacetic acid and their quaternary derivatives have been measured by in vitro and in vivo procedures. The R-enantiomer of dimethylaminoethyl-2-cyclohexyl-2-hydroxy-2-phenylacetate was approximately 100 times more active than the corresponding S-enantiomer both in vivo and in vitro. In contrast, the differences in potencies of the enantiomers of the other compounds were smaller in vivo than in vitro and moreover, the in vivo differences in potency decreased as the potency of the racemates increased. The relevance of these results to general studies of enantiomeric differences is discussed.

Enantioselective open-tubular capillary electrochromatography using cyclodextrin-modified gold nanoparticles as stationary phase

Enantioselective open-tubular CEC (OTCEC) with thiolated β-CD modified gold nanoparticles (CD-GNPs) as stationary phase was developed. The enantioselective OT capillary column was fabricated by electrostatic assembly of poly(diallydimethylammonium chloride) (PDDA) followed by self-adsorption of negatively charged CD-GNPs. The enantioselective capillary column has a steady EOF mobility over a wide pH range of 3.0 to 9.2 (RSD 4.8%), and is quite stable over 240 min with very good column to column reproducibility. Efficient enantioseparation of the presented method was demonstrated by analyzing three drug enantiomers. Our results show that the column exhibits good run-to-run repeatability for enantioseparations and can maintain the enantioselectivity for more than 1 month if the column was stored in CD-GNPs solution at 4 °C.

Stereoselective actions of substituted benzamide drugs on cerebral dopamine mechanisms.

Abstract Apomorphine-induced locomotor activity in reserpine-pretreated mice was antagonized by pretreatment with (-)-sulpiride and (-)-sultopride. The (+)-enantiomers were inactive. Apomorphine- and amphetamine-induced stereotyped behaviour in rats were antagonized by (-)-sultopride but not by the (+)-enantiomer. Neither enantiomer of sulpiride prevented the onset of the stereotyped response. Both (-)-sulpiride and (-)-sultopride induced increases in striatal and mesolimbic HVA and DOPAC concentrations; (+)-sultopride elevated striatal and mesolimbic DOPAC concentrations but not HVA, while (+)-sulpiride had no effect on HVA or DOPAC in either area. Dopamine concentrations were reduced by the enantiomers of sultopride but not by sulpiride. Low concentrations (10−9 −10−66 M) of the (-)-enantiomers of both drugs displaced [3 H]spiperone from its specific binding site in rat striatal preparations, but the (+)-enantiomers were 40 and 100 times less active. However, neither enantiomer of either drug anatagonized the dopamine-induced stimulation of adenylate cyclase in rat striatal preparations. The data suggest that the central pharmacological activity of sulpiride and sultopride resides in the (-)-enantiomers and that this activity occurs at cerebral dopamine receptors not dependent on adenylate cyclase for functional activity.

The central and peripheral activities of anti-acetylcholine drugs. Some concepts of practical relevance.

Abstract Five procedures have been used to compare the pharmacological effects and time-activity profiles of anti-acetylcholine drugs in the central and peripheral nervous system (ens, pns). In particular, the potencies of optically pure enantiomers of anti-acetylcholine drugs which contain an asymmetric centre have been determined. The following four observations are made which are of relevance to all studies of anti-acetylcholine drugs. (1) Times to onset of activity of anti-acetylcholine drugs in three in vivo tests and the duration of action in one in vivo test are shown to increase as affinity constants, determined in vitro, increase. (2) Below a dose of ca 0.03 μmol kg−1no anti-acetylcholine drug produces maximum mydriatic effects and all anti-acetylcholine drugs which have log affinity constant values >9.49 produce maximal effects at approximately this dose. (3) The receptor with which anti-acetylcholine drugs interact is essentially the same in the eye, salivary gland and ens of the mouse, the guinea-pig ileum, and in the cat ens. (4) For any anti-acetylcholine drug the time to onset of effects in the ens is similar to the time of onset of effects in the pns and appears to depend on the affinity constant rather than on the partition properties of the drug. The practical and theoretical significance of these and other observations are discussed.

Critical appraisal for achiral HPLC with polarimetric detection for the determination of drug enantiomers

Critical appraisal for achiral HPLC with polarimetric detection for the determination of drug enantiomers

The stereoselectivity of the ‘single drug binding site’ of human α1-acid glycoprotein (orosomucoid)

The stereoselective binding of six pairs of basic, one pair of acidic drug enantiomers, and one pair of diastereomers for human alpha 1-acid glycoprotein was investigated by means of competition experiments against [3H]propranolol- or [14C]nicardipine-labelled binding sites using equilibrium dialysis to separate free from bound marker ligand. The affinity constants (Ka) for association of [3H]propranolol and [14C]nicardipine with alpha 1-AGP were 1.2 +/- 0.6 X 10(5) M-1 and 3.4 +/- 1.4 X 10(5) M-1, respectively, and control binding amounted to 57 +/- 7 and 91 +/- 2%, respectively. The following selectivity factors, calculated as the ratio of the higher over the lower enantiomer concentrations displacing 15% of control radiomarker binding (IC15-value), were obtained against propranolol and nicardipine: (-)/(+) propranolol: 1.9 and 1.7.; (+)-/(-)-disopyramide: 2.8 and 1.4; (+)-/(-)-verapamil: 1.6 and 1.9; (+)-(S)-/(-)-(R)-202-791, a dihydropyridine derivative: 2.6 and 2.0; (-)-/(+)-asocainol: 1.7 and 3.0; (+)-/(-)-tilidine: 1.1 and approximately equal to 2; (-)-(S)-/(+)-(R)- warfarin: 1.6 and 2.4; (+/-)-cis/(+/-)-trans-trans-tilidine: 1.7 and 1.8. When the calculation of radioligand-free fractions is also taken into account, it is apparent that only the tilidine isomers show no selectivity at propranolol-marked, and the disopyramide isomers at nicardipine-marked alpha 1-AGP-binding sites, in all other cases, a weak selectivity is detectable, which is, however, far below the values obtained for most neurotransmitter receptors. It is concluded that the single drug binding site of alpha 1-AGP is only slightly stereoselective and that the stereoselective binding of the drugs investigated is probably of no clinical consequence.

The Significance of Chirality in Drug Design and Development

Proteins are often enantioselective towards their binding partners. When designing small molecules to interact with these targets, one should consider stereoselectivity. As considerations for exploring structure space evolve, chirality is increasingly important. Binding affinity for a chiral drug can differ for diastereomers and between enantiomers. For the virtual screening and computational design stage of drug development, this problem can be compounded by incomplete stereochemical information in structure libraries leading to a "coin toss" as to whether or not the "ideal" chiral structure is present. Creating every stereoisomer for each chiral compound in a structure library leads to an exponential increase in the number of structures resulting in potentially unmanageable file sizes and screening times. Therefore, only key chiral structures, enantiomeric pairs based on relative stereochemistry need be included, and lead to a compromise between exploration of chemical space and maintaining manageable libraries. In clinical environments, enantiomers of chiral drugs can have reduced, no, or even deleterious effects. This underscores the need to avoid mixtures of compounds and focus on chiral synthesis. Governmental regulations emphasizing the need to monitor chirality in drug development have increased. The United States Food and Drug Administration issued guidelines and policies in 1992 concerning the development of chiral compounds. These guidelines require that absolute stereochemistry be known for compounds with chiral centers and that this information should be established early in drug development in order that the analysis can be considered valid. From exploration of structure space to governmental regulations it is clear that the question of chirality in drug design is of vital importance.

Evaluation of chlorine containing cellulose‐based chiral stationary phases for the LC enantioseparation of basic pharmaceuticals using polar non‐aqueous mobile phases

The discrimination ability of three cellulose-based chiral stationary phases (CSPs) was evaluated towards the enantiomers of basic drugs, using ACN as the main solvent in polar organic mobile phases. The study was focused on CSPs containing cellulose tris(3-chloro-4-methylphenylcarbamate) (3-Cl-4-MePC), cellulose tris(4-chloro-3-methylphenylcarbamate) (4-Cl-3-MePC) or cellulose tris(3,5-dichlorophenylcarbamate) (3,5-diClPC) as the chiral selector. The behaviour of these CSPs was studied systematically in order to investigate the influence of the presence and position of the chlorine substituents on the phenylcarbamate moieties on the retention and resolution of the enantiomers. The evaluation was made with three different generic mobile phases, namely ACN/0.1%DEA/0.1% TFA (DEA, diethylamine), ACN/0.1%DEA/0.2% FA and ACN/0.1%DEA/0.2%AcA, deduced from the previous study. The nature of the acidic additive and of the chiral selector was found to be particularly important for the retention and enantioresolution of these basic compounds. High-resolution values could be obtained for most studied enantiomers with these CSPs, clearly demonstrating the interest of using them in combination with polar organic mobile phases. However, significant differences in enantioresolution between the CSPs have been observed for many compounds, indicating that these phases seem to be quite complementary.

ChemInform Abstract: Pharmacologically Active Compounds in the Environment and Their Chirality

AbstractReview: 212 refs.

Effects on enantiomeric drug disposition and open-field behavior after chronic treatment with venlafaxine in the P-glycoprotein knockout mice model

P-glycoprotein (P-gp) plays an important role in the efflux of drugs from the brain back into the bloodstream and can influence the pharmacokinetics and pharmacodynamics of drug molecules. To our knowledge, no studies have reported pharmacodynamic effects of any antidepressant drug in the P-gp knockout mice model. The aim of this study was to investigate the enantiomeric venlafaxine and metabolite concentrations in serum and brain of abcb1ab⁻/⁻ mice compared to wild-type mice upon chronic dosing, and to assess the effect of venlafaxine treatment on open-field behavior. P-gp knockout and wild-type mice received two daily intraperitoneal injections of venlafaxine (10 mg/kg) over ten consecutive days. Locomotor and rearing activities were assessed on days 7 and 9. After 10 days, drug and metabolite concentrations in brain and serum were determined using an enantioselective LC/MS/MS method. The brain concentrations of venlafaxine and its three demethylated metabolites were two to four times higher in abcb1ab⁻/⁻ mice compared to abcb1ab+/+ mice. The behavioral results indicated an impact on exploration-related behaviors in the open-field as center activity was increased, and rears were decreased by venlafaxine treatment. Our results show that P-gp at the blood-brain barrier plays an important role in limiting brain entry of the enantiomers of venlafaxine and its metabolites after chronic dosing. Taken together, the present pharmacokinetic and pharmacodynamic findings offer the possibility that the expression of P-gp in patients may be a contributing factor for limited treatment response.

Bioequivalence of Venlafaxine Modified-Release Capsule Revisited with an Innovative Approach Using Experimental and Predictive Models

To evaluate the venlafaxine:O-desmethyl venlafaxine (active metabolite) in vivo formation ratio (MR) in three independent bioequivalence (BE) studies consisting of single-dosed (under fasted and fed conditions) and multiple-dosed clinical trials on healthy subjects. The pooled data pharmacokinetic (PK) analysis demonstrates a model to conduct enantiomer/racemate/active metabolite bioanalysis for regulatory submission of bioavailability/bioequivalence (BA/BE) studies using an interesting MR concept. BE was established for all three studies. Moreover, the venlafaxine:O-desmethyl venlafaxine MR for C(max) and AUC(last) differed by more than 50% for fasted and fed single-dosed studies, while pooled data analysis found the MR for C(max) to be approximately 0.63 and the AUC to be approximately 0.36 for both test and reference drugs. However, negligible variation was observed for both rate and extent of drug and active metabolite absorption into the systemic circulation at steady state, as the MR for both C(max) and AUC was approximately 0.62. The applications/consequences of the above results are immense. First, an achiral assay for venlafaxine and O-desmethyl venlafaxine estimation in human plasma has been justified for the regulatory acceptance of BA/BE studies, supported with both single- and multiple-dosed PK data showing negligible variation in terms of MR at C(max). Second, the current investigation shows the MR to be within ±10% when compared with the single-dosed reported study on a western population. Third, the racial effect would not lead to any significant clinical outcome using an interchangeable venlafaxine 150-mg capsule manufactured by Ranbaxy with an Efexor 150-mg capsule manufactured by Wyeth. Furthermore, a decision tree is proposed to evaluate if a racemate or an enantiomer drug and active metabolite bioanalysis should be executed for BA/BE regulatory submission using respective achiral or chiral assays when the drug moiety is a racemate or an enantiomer, formulated in modified-release dosage forms.

Chiral Drugs: Current Status of the Industry and the Market

The biological activity of chiral substances often depends upon their stereochemistry, since the living body is a highly chiral environment. A large percentage of commercial and investigational pharmaceutical compounds are enantiomers, and many of them show significant enantioselective differences in their pharmacokinetics and pharmacodynamics. The importance of chirality of drugs has been increasingly recognized, and the consequences of using them as racemates or as enantiomers has been frequently discussed in the pharmaceutical literature during recent years. Most of the chiral drugs are administered as racemates, despite the fact that the optical isomers of a racemic drug can exhibit different pharmacological profiles in living systems. These differences can be expressed in e.g. the affinity of the enantiomers for certain receptor subtypes or enzymes, distribution rates, their metabolism and excretion, in antagonistic actions relative to each other, or their toxicological properties. Obviously, the more chiral centres present in a (drug) molecule, the more complex the situation becomes. The origin of the discovery of the role of stereochemistry in biochemical environments dates back to the late 1850’s, when Pasteur reported the different destruction rates of dextro and levo ammonium tartrate by the mold Penicillium glaucum. In the conclusion of his observations, Pasteur wrote: “Most natural organic products, the essential products of life, are asymmetric and possess such asymmetry that they are not superimposable on their images.” For some reason, this knowledge of racemic organic compounds and racemisation seemed to be forgotten until the question of racemic compounds was raised by Ariens in the late 1980’s. He asked the question” why we in some cases have to give doses to the patient where half of the content has no effect or the opposite effect?”. After this rediscovery of stereochemistry, the regulatory authorities defined more strict requirements on drug discovery and chiral compounds. Besides the ethical reasons, the therapeutic benefit (efficacy and safety) and, in several instances, extension of the life cycle of drugs have been the motivation for developing single enantiomers. Single–enantiomer drug sales show a continuous growth worldwide and many of the topselling drugs are marketed as single enantiomers. In this context, there has been a rapid development of enantioselective synthetic methodologies, which have now reached a high degree of diversity and complexity. Simultaneously, this new trend produced a rapid increase in the demand for stereoselective separation techniques and analytical assays for precise determination of the enantiomeric purity of chiral compounds. The development of chiral stationary phases (CSPs) or chiral selectors for gas chromatography (GC), liquid chromatography (LC) and capillary electrophoresis (CE) rapidly opened a new dimension in the area of separation technologies. The business of developing single isomer drugs came about, because the chemical production methods used for pharmaceuticals often produced racemic mixtures of two enantiomers. In the case of thalidomide, it was shown that one enantiomer was responsible for efficacy and another for side effects. Worldwide, the market for chiral fine chemicals sold as single enantiomers was $6.63 billion in 2000 and is expected to grow at a rate of 13.2% annually, reaching $16.0 billion in 2007. The drug industry is the driving engine for this strong growth, accounting for 81.2% of the total, equal to an overall worth of $5.38 billion, in the year 2000. The remaining $1.25 billion is divided among Copyright © 2007 by School of Pharmacy Shaheed Beheshti University of Medical Sciences and Health Services Iranian Journal of Pharmaceutical Research (2007), 6 (2): 73-74

ChemInform Abstract: Resolution of Chiral Drugs

Many drugs are administered as racemates, yet the enantiomers may have important pharmacodynamic and/or pharmacokinetic differences. Chiral analytical techniques are necessary to properly assess and understand such differences. Novel techniques to separate drug enantiomers present in serum and/or urine have recently emerged. This review describes commonly used approaches to chiral drug analysis in: diastereomeric derivatization, chiral mobile-phase additives, chiral stationary phases, and enantioselective immunoassays. Each of these techniques has advantages and disadvantages and no single approach has emerged as the method of choice.

Serum Concentrations of Citalopram – Dose-Dependent Variation inR- andS-Enantiomer Ratios

The antidepressive effect of racemic citalopram (CIT) is exerted by S-CIT, while R-CIT is a partial antagonist to S-CIT. Since R-and S-CIT are metabolized by different pathways, we investigated whether the ratio of S- and R-CIT may differ between individuals on the same dose of racemic CIT, and if a possible variability in the R/S-ratio could be dose-dependent. A chiral analysis of R- and S-CIT in serum samples taken from 88 female patients receiving treatment with racemic CIT was performed using high-pressure liquid chromatography. The mean levels of R-CIT were significantly higher than those of S-CIT in all dose groups. The R/S-CIT ratio increased from 1.99 to 2.45 with an increase in the dose (p<0.05), and the interindividual variance in the R/S-CIT ratio was up to four-fold on the same dosage. Our findings show that the stereoselective metabolism of citalopram IN VIVO has pharmacokinetic consequences reflected by dose dependent variations of enantiomeric drug concentrations, as well as substantial interindividual variabilities in the ratios of the concentrations. The clinical consequences, however, are unclear and should be further explored.

Blood–brain barrier penetration of the enantiomers of venlafaxine and its metabolites in mice lacking P-glycoprotein

According to in vitro studies the enantiomers of venlafaxine display different degrees of serotonin and noradrenaline reuptake inhibition. Therefore, clarification of the enantiomeric drug distribution between serum and brain is highly warranted. To elucidate if P-glycoprotein (P-gp) in a stereoselective manner transports venlafaxine and its metabolites out of the brain we used abcb1ab double-knockout mice that do not express P-gp. A single dose of racemic venlafaxine (10 mg/kg bw) was intraperitoneally injected to knockout (-/-) and wildtype (+/+) mice. Serum and brain samples were collected 1, 3, 6 and 9 h following drug administration for analysis by LC/MS/MS. One to six hours post-dose, the brain concentrations of venlafaxine, O-desmethylvenlafaxine and N-desmethylvenlafaxine were 2-3, 2-6 and 3-12 times higher in abcb1ab (-/-) mice compared to abcb1ab (+/+) mice, respectively. No major differences in the serum and brain disposition of the S- and R-enantiomers of venlafaxine and its metabolites were found between the groups. We conclude that P-gp decreases the penetration of the S- and R-enantiomers of venlafaxine and its major metabolites into the brain. No evidence of a stereoselective P-gp mediated transport of these substances was observed.