Molecular Micelles Research Articles

Surface charge switchable fluorinated small molecular micelles for enhanced photodynamic therapy for bacterial infections

Photodynamic therapy (PDT) employs reactive oxygen species (ROS) from a photosensitizer (PS) under light, inhibiting multi-drug resistance in bacteria. However, hypoxic conditions in infection sites and biofilms challenge PDT efficiency. We developed fluorinated small molecular micelles (PF-CBMs) as PS carriers to address this, relieving hypoxia and enhancing PS penetration into biofilms. Perfluorocarbons in PF-CBMs transport more oxygen due to their excellent oxygen-dissolving capability. Fluorination enhances loading capacity and serum stability, reduces premature release, and improves cellular uptake, to improve PDT efficacy. PF-CBMs, with acid-induced surface charge transformation, exhibit superior biofilm penetration, resulting in increased antibiofilm activity of PDT. Compared to fluorine-free micelles (PC-CBMs), PF-CBMs demonstrate better serum stability, higher drug loading, and reduced premature release, leading to significantly improved antibacterial efficacy in vitro and in vivo. In conclusion, fluorinated micelles with surface charge reversal enhance PDT for antibacterial and antibiofilm applications.

Update on chiral recognition mechanisms in separation science.

The stereospecific analysis of chiral molecules is an important issue in many scientific fields. In separation sciences, this is achieved via the formation of transient diastereomeric complexes between a chiral selector and the selectand enantiomers driven by molecular interactions including electrostatic, ion-dipole, dipole-dipole, van der Waals or π-π interactions as well as hydrogen or halogen bonds depending on the nature of selector and selectand. Nuclear magnetic resonance spectroscopy and molecular modeling methods are currently the most frequently applied techniques to understand the selector-selectand interactions at a molecular level and to draw conclusions on the chiral separation mechanism. The present short review summarizes some of the recent achievements for the understanding of the chiral recognition of the most important chiral selectors combining separation techniques with molecular modeling and/or spectroscopic techniques dating between 2020 and early 2024. The selectors include polysaccharide derivatives, cyclodextrins, macrocyclic glycopeptides, proteins, donor-acceptor type selectors, ion-exchangers, crown ethers, and molecular micelles. The application of chiral ionic liquids and chiral deep eutectic solvents, as well as further selectors, are also briefly addressed. A compilation of all published literature on chiral selectors has not been attempted.

Self‐Limited Hyperbranching Polymerization of Di‐N‐Phenoxycarbonyl N‐Substituted Diglycines

AbstractHyperbranched poly(α‐amino acid)s have caught great attention due to their excellent biocompatibility, biodegradability, and bioactivity. However, hyperbranched polypeptoids are rarely reported because of the challenge of synthesis. Herein, unique polymerizations of N,N'‐diphenoxycarbonyl‐N,N'‐(1,4‐phenylenebismethylene)–diglycine (di‐NNPC, an A*B*‐A*B*‐type monomer) which uncommonly result in hyperbranched polypeptoids rather than gels after the complete consumption of di‐NNPC are reported. The molecular weights of the hyperbranched polypeptoids are controlled by the degree of branching via tuning the initial concentrations of monomer and basic catalyst. N‐Phenyloxycarbonyl glycine pendant groups plentifully remained on the polymer chains are used for Gd3+ complexation and post‐modification, which makes the hyperbranched polypeptoids multifunctional single molecular micelles applicable as magnetic resonance imaging contrast agents and photoluminescence probes.

Antibiotic-based small molecular micelles combined with photodynamic therapy for bacterial infections

The appearance of multidrug-resistant bacteria and the formation of bacterial biofilms have necessitated the development of alternative antimicrobial therapeutics. Antibiotics conjugated with or embedded in nano-drug carriers show a great potential and advantage over free drugs, but the mass proportion of carriers generally exceeds 90% of the nano-drug, resulting in low drug loading and limited therapeutic output. Herein, we fabricated a nanocarrier using antibiotics as the building blocks, minimizing the use of carrier materials, significantly increasing the drug loading content and treatment effect. Firstly, we conjugated betaine carboxylate with ciprofloxacin (CIP) through an ester bond to form the amphiphilic conjugate (CIP-CB), which self-assembled into micelles (CIP-CBMs) in aqueous solutions, with a CIP loading content as high as 65.4% and pH-induced surface charge reversal properties. Secondly, a model photosensitizer (5, 10, 15, 20-tetraphenylporphyrin (TPP)) was encapsulated in CIP-CBMs, generating infection-targeted photodynamic/antibiotic combined nanomedicines (denoted as TPP@CIP-CBMs). Upon accumulation at infection sites or in deep bacterial biofilms, the ester bond between the betaine carboxylate and CIP is cleaved to release free TPP and CIP, leading to a synergetic antibacterial and antibiofilm activity in vitro and in vivo.

Capillary Electrophoresis Mass Spectrometry: Developments and Applications for Enantioselective Analysis from 2011-2020.

It is now more than 25 years since the first report of enantioselective analysis by capillary electrophoresis-mass spectrometry (CE-MS) appeared. This article reviews the power of chiral CE-MS in resolving issues on the use of chiral selector incompatibility with MS and poor detectability encountered for chiral compounds by UV detection. The review begins with the general principles, requirements, and critical aspects of chiral CE-MS instrumentation. Next, the review provides a survey of MS-compatible chiral selectors (CSs) reported during the past decade, and the key achievements encountered in the time period using these CSs. Within the context of the strategies used to combine CE and MS, special attention is paid to the approaches that feature partial filling technique, counter-migration techniques, and direct use of CS, such as molecular micelles. In particular, the development and application of moving and fixed CS for EKC-MS, MEKC-MS, and CEC-MS demonstrate how various chiral compounds analyses were solved in a simple and elegant way during the 2010–2020 review period. The most noteworthy applications in the determination of chiral compounds are critically examined. The operating analytical conditions are detailed in the Tables, and the authors provide commentary on future trends of chiral separations by CE-MS.

Surface Charge Switchable Antibiotics Based Small Molecular Micelles Combined with Photodynamic Therapy for Antibacterial and Antibiofilm Applications

Surface Charge Switchable Antibiotics Based Small Molecular Micelles Combined with Photodynamic Therapy for Antibacterial and Antibiofilm Applications

Helical Self-Assembly of Amphiphilic Chiral Azobenzene Alternating Copolymers.

Imposing chirality to supramolecular architectures is an important step forward toward understanding and utilization of chiral nanomaterials. This article reports the self-assembly of amphiphilic chiral alternating copolymers of poly(binaphthyl azobenzene-alt-hexaethylene glycol) (P(BNPAzo-alt-EG6)) into helical supramolecular rods. Unlike conventional chiral assembly of copolymers largely through intermolecular organization, the intrachain stacking of chiral units along the main chain into single molecular micelles with amplified axial chirality of binaphthyl is key to the formation of helical supramolecular rods, which takes advantage of the particular chiral unit and soft unit alternating topological structure of the backbones. Moreover, the supramolecular self-assembly is light reversible because the azobenzene rings in the backbone scarcely execute trans- to cis-isomerization upon UV irradiation, and therefore the supramolecular rods keep their sublevel chirality even though the helical appearance was destroyed. This work paves an effective route to construct and regulate chiral supramolecular architectures and reveals an insight into natural and artificial chiral self-assembly.

Chiral Recognition of Dansyl Derivatives with an Amino Acid-Based Molecular Micelle: A Molecular Dynamics Investigation

In this study, the chiral separation mechanisms of Dansyl amino acids, including Dansyl-Leucine (Dans-Leu), Dansyl-Norleucine (Dans-Nor), Dansyl-Tryptophan (Dans-Trp) and Dansyl-Phenylalanine (Dans-Phe) binding to poly-sodium N-undecanoyl-(L)-Leucylvalinate, poly(SULV), were investigated using molecular dynamics simulations. Micellar electrokinetic chromatography (MEKC) has previously shown that when separating the enantiomers of these aforementioned Dansyl amino acids, the L- enantiomers bind stronger to poly(SULV) than the D- enantiomers. This study aims to investigate the molecular interactions that govern chiral recognition in these systems using computational methods. This study reveals that the computationally-calculated binding free energy values for Dansyl enantiomers binding to poly(SULV) are in agreement with the enantiomeric order produced in experimental MEKC studies. The L- enantiomers of Dans-Leu, Dans-Nor, Dans-Trp, and Dans-Phe binding to their preferred binding pockets in poly(SULV) yielded binding free energy values of -21.8938, -22.1763, -21.3329 and -13.3349 kJ·mol-1, respectively. The D- enantiomers of Dans-Leu, Dans-Nor, Dans-Trp, and Dans-Phe binding to their preferred binding pockets in poly(SULV) yielded binding free energy values of -14.5811, -15.9457, -13.6408, and -12.0959 kJ·mol-1, respectively. Furthermore, hydrogen bonding analyses were used to investigate and elucidate the molecular interactions that govern chiral recognition in these molecular systems.

Establishing repeatability and ruggedness of chiral separations in micellar electrokinetic chromatography mass spectrometry: Combined use of covalently bonded capillary column and molecular micelles

Although micellar electrokinetic chromatography-mass spectrometry (MEKC-MS) using bare silica capillary filled with molecular micelles is an advantageous hyphenated technique for chiral analysis, it is still in the developmental stage. This is mainly because of the lower repeatability of retention time and peak area associated with the difficulty in controlling electroosmotic flow on bare silica capillaries. Besides the lower robustness and electrospray erosion of the fused-silica capillary tip, the lifetime is limited for 10–15 runs per capillary column. We have tested a new MEKC-MS method to eradicate this problem using a covalently bonded 2-acrylamido-2-methyl-1-propane-sulfonic acid (AMPS) column filled with free floating molecular micelles, polysodium N-undecenoxy carbonyl-L-leucinate (poly-L-SUCL). Simultaneous enantiomeric separations and MS/MS detection of three β-blockers [atenolol (ATEN), metoprolol (METO) and, pindolol (PINDO)] was achieved within 25 min with improved column lifetime of at least 45–50 runs. Excellent repeatability of retention time was observed for β-blockers, as evidenced by the relative standard deviation of less than 2% and 3% for intra-capillary and inter-capillary column, respectively. The linear calibration range of both β-blockers was simultaneously established using enantiomers of PINDO as an internal standard. The limit of detection and the lower limit of quantitation were 0.2 μg/mL and 0.5 μg/mL, respectively, for both ATEN and METO. Acceptable recovery of the enantiomeric content of the commercial METO tablet (98-99.5%) and ATEN tablet (89-92.5%) were obtained with adequate system suitability for the precision of peak area (≤10% RSD) under optimum conditions. The developed MEKC-MS approach was extended, which provided broader repeatibility of chiral separation to a panel of primary, secondary and tertiary amines as well as one anionic chiral compound.

Carbohydrate-Based Polymeric Surfactants for Chiral Micellar Electrokinetic Chromatography (CMEKC) Coupled to Mass Spectrometry.

Polymeric surfactants (molecular micelles, MoMs) with a variety of chiral head groups and chain lengths may be the most promising chiral selectors when used for sensitive detection of chiral compounds in micellar electrokinetic chromatography-mass spectrometry (MEKC-MS). Various carbohydrate-based MS compatible surfactants with phosphate and sulfate head groups have been recently synthesized in our laboratory for its application in CMEKC-MS. In this chapter, we illustrate that the synthesized glucopyranoside-based MoMs are fully compatible with electrospray ionization MS and can be successfully used as a chiral selector for high-throughput screening of multiple chiral compounds using MRM mode in CMEKC-MS/MS experiments. This chapter describes in detail synthesis and utility of α- and β-glucopyranoside-based polymeric surfactant with two different chain lengths and head groups. The presented examples optimize the effect of appropriate millimolar concentration of monomer sugar surfactants required for polymerization as it affects the separations of acidic and basic compounds. Under the optimized concentration of the monomer needed for polymerization (i.e., an equivalent monomer concentration of MoMs), the superiority of MEKC-MS over MEKC-UV is evident. Structurally similar basic drugs with the difference in hydrophobicity are first tested in MEKC-MS to find the optimum head group and optimum chain length with the aim for developing a widely applicable polymeric glucopyranoside-based surfactant. The partial enantioresolution of several structurally similar basic compounds is significantly improved when switching from one head group to another head group of the glucopyranoside MoMs. Thus, complementary separations using poly-N-β-D-SUGP versus poly-N-β-D-SUGS were seen. This phenomenon also exists when comparing the MoMs, which differ in an anomeric configuration such as poly-N-α-D-SUGP and poly-N-β-D-SUGP.

Characterization of Amino Acid Based Molecular Micelles with Molecular Modeling.

The enantiomers of chiral drugs often have different potencies, toxicities, and biochemical properties. Therefore, the FDA and other worldwide regulatory agencies require manufactures to test and prove the enantiomeric purity of chiral drugs. Amino acid based molecular micelles (AABMM) have been used in chiral CE separations since the 1990's because of their low environmental impact and because their properties can easily be tuned by changing the amino acids in the chiral surfactant headgroups. Using molecular dynamics simulations to investigate the structures and properties of AABMM is part of an ongoing study focusing on investigating and elucidating the factors responsible for chiral recognition with AABMM. The results will be useful for the proper design and selection of more efficient chiral selectors. The micelles investigated contained approximately twenty covalently linked surfactant monomers. Each monomer was in turn composed of an undecyl hydrocarbon chain bound to a dipeptide headgroup containing of all combinations of L-Alanine, L-Valine, and L-Leucine. These materials are of interest because they are effective chiral selectors in capillary electrophoresis separations. Molecular dynamics simulation analyses were used to investigate how the sizes and positions of the headgroup amino acid R-groups affected the solvent accessible surface areas of each AABMM chiral center. In addition, headgroup dihedral angle analyses were used to investigate how amino acid R-group size and position affected the overall headgroup conformations. Finally, distance measurements were used to study the structural and conformational flexibilities of each AABMM headgroup. All analyses were performed in the context of a broader study focused on developing structure-based predictive tools to identify the factors responsible for (a) self-assembly, (b) function, (c) higher ordered structure and (d) molecular recognition of these amino acid based molecular micelles.

DOTA Functionalized Cross-Linked Small-Molecule Micelles for Theranostics Combining Magnetic Resonance Imaging and Chemotherapy.

Herein, a new theranostic nanoplatform was built by introducing DOTA on the surface of the cross-linked small molecular micelles. This nanoplatform consisted of three parts: (1) Hydrophobic alkyl chain functioned with alkenes as cross-linkable part to ensure the robust stability in vivo; (2) Hydrophilic DOTA for not only gadolinium ions complexion as MR imaging contrast agents but also platinum ions complexion as drug carriers; (3) Cisplatin as antitumor drug and MR imaging amplifier simultaneously. The metal loading content of the resulting Pt/Gd@DOTA-CSMs was up to 12 and 14 wt % for Gd(III) and Pt(II), respectively. The longitudinal relaxation rate of Gd@DOTA-CSMs (8.46 mM-1 s-1) was 2.6 times than that of the clinical Gd-DTPA (3.23 mM-1 s-1). When cisplatin was involved, not only great antitumor efficiency was achieved, but higher longitudinal relaxation rate of Pt/Gd@DOTA-CSMs (11.17 mM-1 s-1) was carried out due to the extension of rotational correlation time. The experiments of Balb/c mice bearing 4T1 xenograft indicated that Pt/Gd@DOTA-CSMs had better antitumor activity, lower side effect than clinically used cisplatin, and greater MR imaging than Gd-DTPA. All the results proved that the nanoplatform of DOTA-CSMs held remarkable potential in the field of therapeutics.

Using molecular dynamics simulations to identify the key factors responsible for chiral recognition by an amino acid-based molecular micelle

Molecular dynamics (MD) simulations were used to investigate the binding of six chiral compounds to the amino acid-based molecular micelle (MM) poly-(sodium undecyl-(L)-leucine-leucine) or poly(SULL). The MM investigated is used as a chiral selector in capillary electrophoresis. The project goal was to characterize the chiral recognition mechanism in these separations and to move toward predictive models to identify the best amino acid-based MM for a given separation. Poly(SULL) was found to contain six binding sites into which chiral compounds could insert. Four sites had similar sizes, shapes, and electrostatic properties. Enantiomers of alprenolol, propranolol, 1,1′-bi-2-naphthyl-2,2′-diyl hydrogen phosphate, 1,1′-bi-2-naphthol, chlorthalidone, or lorazepam were separately docked into each binding pocket and MD simulations with the resulting intermolecular complexes were performed. Solvent-accessible surface area calculations showed the compounds preferentially associated with binding sites where they penetrated into the MM core and shielded their non-polar atoms from solvent. Furthermore, with five of the six compounds the enantiomer with the most favorable free energy of MM association also experienced the most favorable intermolecular hydrogen bonding interactions with the MM. This result suggests that stereoselective intermolecular hydrogen bonds play an important role in chiral discrimination in separations using amino acid-based MMs.GRAPHICAL ABSTRACT

Supramolecular Chemistry of Cobalt-Corrins

Cobalt corrins are chiral relatives of the cobalt porphyrins, in which the ligand is much less unsaturated and is also ‘contracted’ by one carbon. This type of ligand strongly affects the redox and coordination chemistry of the bound cobalt center. Some lipophilic artificial cobalt corrins have been prepared in tremendous synthetic efforts in the Eschenmoser labs. The natural analogues offer an apparently more straight forward chemical entry to cobalt corrins. However, the natural cobalt corrins are all hydrophilic and highly substituted at their periphery, and they are distantly related to heme and chlorophyll. Although these cobalt corrins are very soluble in water, their natural roles are intimately connected with their essential and rich organometallic chemistry, which is all cobalt-centered.[1] In this lecture, means for specific supramolecular assemblies based on modified natural cobalt corrins are delineated, as well as consequences of the supramolecular restructuring on the organometallic and redox-chemistry. So far, in the excursions to be reported, the cobalt corrin core is not altered and its redox and spectral properties are largely conserved. A unique basis for specific supramolecular assemblies is then provided by the (organometallic) ligand itself, either, when attached to and bridging more than one metal center, or, when providing a structuring interface itself, for the build-up of supra-molecules. In a second general type of supramolecular assemblies the functional groups attached at the peripheral substituents of the natural corrinoids are used in a uniform way. By this means, the direct environment of the cobalt corrin moiety may be affected and structured in a global fashion, generating, e.g., a type of molecular micelles. A third type of supramolecular assembly makes selective use of single sites of the complex, but largely conserved sphere of the peripheral substituents, in order to generate specifically structured cobalt-corrins and their dimers. These latter ones may feature binuclear (organometallic) coordination chemistry, which would allow for synchronized redox processes in a highly structured, ‘second’ sphere around the metal centers. I thank the Austrian Science Foundation (FWF, project P-28892 and earlier projects) for generous financial support.

Spectroscopic properties of an amphiphilic naphthalimide derivative with high surface activity and detection of trace salt in the water as a fluorescent sensor

A novel amphiphilic fluorescent sensor bearing high surface activity, 4-(3-Dimethyl-benzyl propylamine ammonium)-N-dodecyl-1,8-naphthalimide chloride (DADNC), has been designed and used to rapidly detect trace salt in the pure water as an “on-off” fluorescent sensor. The surface activity and high sensitivity to trace salt ions and organic contaminants were measured. The average particle sizes of the molecular micelles of DADNC were 31–55nm in diameter. The fluorescence of DADNC is highly sensitive to trace salt ions and ionic organic contaminants. The aggregation and disintegration of DADNC in the presence of ionic micelles showed sensitive “on-off-on” fluorescence switching. There were good linear relationships between its fluorescence intensities and the salt concentrations at very low salt concentrations from 1×10^−8 to 1×10^−5mol/L. This system could be used as an “on-off” fluorescence sensor for the detection and the quantitative determination of ionic substrate or anionic organic compounds at ultralow ion concentration. It has potential applications in the recognition and detection of the trace amounts of the salt ions and organic contaminants in the drinking water and pure water.

Investigation of chiral recognition by molecular micelles with molecular dynamics simulations

ABSTRACTMolecular dynamics simulations were used to characterize the binding of the chiral drugs chlorthalidone and lorazepam to the molecular micelle poly-(sodium undecyl-(L)-leucine-valine). The project’s goal was to characterize the nature of chiral recognition in capillary electrophoresis separations that use molecular micelles as the chiral selector. The shapes and charge distributions of the chiral molecules investigated, their orientations within the molecular micelle chiral binding pockets, and the formation of stereoselective intermolecular hydrogen bonds with the molecular micelle were all found to play key roles in determining where and how lorazepam and chlorthalidone enantiomers interacted with the molecular micelle.

Nanostructure characterization of asphalt-aggregate interface through molecular dynamics simulation and atomic force microscopy

Although it has been widely accepted that the asphalt-aggregate interface plays as one of the most important roles for the durability of asphalt concrete, it is still a big challenge to elucidate the failure mechanisms when it is exposed to moisture environment due to the lack of sufficient information on the nanostructure of the asphalt-aggregate interfaces. To understand the failure mechanisms of the asphalt concrete resulted from the moisture damage, the Molecular Dynamic (MD) simulation and the PeakForce Quantitative Nanomechanics atomic force microscopy (PFQN-AFM) were used to characterize the nanostructure of the asphalt-aggregate interface. The MD simulation results demonstrated that, at the asphalt-aggregate interface, the nanostructure is constructed by periodical molecular bulgy boundaries and central cavities. The polar aromatics not only form the structural skeleton with the asphaltenes aggregations, but also act as the stabilizer to enhance the compatibility between asphaltenes and maltenes. Besides, it was found that the intermolecular gaps can be filled by naphthene aromatics, and molecular micelles can be lubricated by saturates. The AFM results show that, some regions in the asphalt-aggregate interface have lower adhesion force with the tip than the surrounding regions. The low adhesion regions are relevant to the structure that polar molecules aggregate into boundaries and evade the center. The dissolution of polar molecular into water induces the nanostructure collapse, and further leads to the asphalt-aggregate interface debonding. This study not only presents the nanostructure of the asphalt-aggregates interfaces, but also helps to understand the moisture damage mechanisms of the asphalt concrete.

Chiral Capillary Electrophoresis-Mass Spectrometry: Developments and Applications in the Period 2010-2015: A Review.

The sensitive detection of chiral compounds by capillary electrophoresis (CE) in biological samples remains a significant challenge and is currently considered a bottleneck in many chiral analysis projects. Chiral CE-MS can significantly improve the limit of detection and provide high sensitivity compared with chiral CE-UV. Chiral selectors such as modified cyclodextrins (CDs) and polymeric surfactants (a.k.a. molecular micelles, MoMs) in electrokinetic chromatography (EKC), micellar electrokinetic chromatography (MEKC) and capillary electrochromatography (CEC) have been developed to address the need for high sensitivity by CE-MS. However, several problems remain to be investigated to fully understand the potential of these hyphenation modes. This review provides introduction to major chiral CE-MS modes for the novice and highlights the important working principles of each mode of chiral CE-MS. Next, recent practical developments and progress in chiral CE-MS dating from January 2010 to September 2015 are described. The achievements in clinical and biomedical sciences using a variety of chiral selectors such as CDs and MoMs in EKC-MS, MEKC-MS and CEC-MS are discussed. Finally, conclusions and future prospects of CE-MS in chiral analysis are drafted.

Molecular dynamics simulation and NMR investigation of the association of the β-blockers atenolol and propranolol with a chiral molecular micelle

Molecular dynamics simulations and NMR spectroscopy were used to compare the binding of two β-blocker drugs to the chiral molecular micelle poly-(sodium undecyl-(L)-leucine-valine). The molecular micelle is used as a chiral selector in capillary electrophoresis. This study is part of a larger effort to understand the mechanism of chiral recognition in capillary electrophoresis by characterizing the molecular micelle binding of chiral compounds with different geometries and charges. Propranolol and atenolol were chosen because their structures are similar, but their chiral interactions with the molecular micelle are different. Molecular dynamics simulations showed both propranolol enantiomers inserted their aromatic rings into the molecular micelle core and that (S)-propranolol associated more strongly with the molecular micelle than (R)-propranolol. This difference was attributed to stronger molecular micelle hydrogen bonding interactions experienced by (S)-propranolol. Atenolol enantiomers were found to bind near the molecular micelle surface and to have similar molecular micelle binding free energies.

A molecular dynamics simulation study of the association of 1,1′-binaphthyl-2,2′-diyl hydrogenphosphate enantiomers with a chiral molecular micelle

Molecular dynamics (MD) simulations were used to investigate the binding of 1,1′-binaphthyl-2,2′-diyl hydrogenphosphate (BNP) enantiomers to the molecular micelle poly-(sodium undecyl-(L,L)-leucine-valine) (poly(SULV)). Poly(SULV) is used as a chiral selector in capillary electrophoresis separations. Four poly(SULV) binding pockets were identified and either (R)-BNP or (S)-BNP were docked into each pocket. MD simulations were then used to identify the preferred BNP binding site. Within the preferred site, both enantiomers formed hydrogen bonds with poly(SULV) and penetrated into the poly(SULV) core. Comparisons of BNP enantiomer binding to the preferred poly(SULV) pocket showed that (S)-BNP formed stronger hydrogen bonds, moved deeper into the binding site, and had a lower poly(SULV) binding free energy than the (R) enantiomer. Finally, MD simulation results were in agreement with capillary electrophoresis and NMR experiments. Each technique showed (S)-BNP interacted more strongly with poly(SULV) than (R)-BNP and that the site of chiral recognition was near the poly(SULV) leucine chiral center.