In Vitro and In Vivo Sequestration of Methamphetamine by a Sulfated Acyclic CB[n]-Type Receptor

We report the synthesis of two new acyclic sulfated acyclic CB[n]-type receptors ( TriM0 and Me₄TetM0 ) and investigations of their binding properties toward a panel of drugs of abuse ( 1 – 13 ) by a combination of ¹H NMR spectroscopy and isothermal titration calorimetry. TetM0 is the most potent receptor with K_a≥10⁶ M⁻¹ toward methamphetamine, fentanyl, MDMA and mephedrone. TetM0 is not cytotoxic toward HepG2 and HEK 293 cells below 100 μM according to MTS metabolic and adenylate kinase release assays and is well tolerated in vivo when dosed at 46 mg kg⁻¹. TetM0 does not inhibit the hERG ion channel and is not mutagenic based on the Ames fluctuation test. Finally, in vivo efficacy studies show that the hyperlocomotion of mice treated with methamphetamine can be greatly reduced by treatment with TetM0 up to 5 minutes later. TetM0 has potential as a broad spectrum in vivo sequestrant for drugs of abuse.     

The cucurbit[n]uril family

In 1981, the macrocyclic methylene-bridged glycoluril hexamer (CB[6]) was dubbed "cucurbituril" by Mock and co-workers because of its resemblance to the most prominent member of the cucurbitaceae family of plants--the pumpkin. In the intervening years, the fundamental binding properties of CB[6]-high affinity, highly selective, and constrictive binding interactions--have been delineated by the pioneering work of the research groups of Mock, Kim, and Buschmann, and has led to their applications in waste-water remediation, as artificial enzymes, and as molecular switches. More recently, the cucurbit[n]uril family has grown to include homologues (CB[5]-CB[10]), derivatives, congeners, and analogues whose sizes span and exceed the range available with the alpha-, beta-, and gamma-cyclodextrins. Their shapes, solubility, and chemical functionality may now be tailored by synthetic chemistry to play a central role in molecular recognition, self-assembly, and nanotechnology. This Review focuses on the synthesis, recognition properties, and applications of these unique macrocycles.     

IBX Oxidation of Benzenedimethanols in the Presence of Cucurbit[8]uril

The cucurbit[8]uril (Q[8]) mediated oxidation of benzenedimethanols with o‐iodoxybenzoic acid (IBX) in aqueous solution has been investigated, and the results reveal the supramolecular catalysis depends on the electronic and geometric structure of substrate. In the cases of o‐benzenedimethanol ( 1a ) and m‐benzenedimethanol ( 1b ), the IBX oxidation could be obviously enhanced by the addition of Q[8] at different extent. There is no observation of the catalytic activity of Q[8] when p‐benzenedimethanol ( 1c ) is subjected to the IBX oxidation. The addition amount of Q[8] is discussed herein, and the addition of more than 10% mol catalyst cannot improve the oxidation much more. The investigation of host‐guest interactions by isothermal titration calorimetry implies the supramolecular catalysis is related to the formation of complexes between benzenedimethanols and cucurbit[8]uril.     

Complex Formation of Crown Ethers with the Cucurbit[6]uril–Spermidine and Cucurbit[6]uril–Spermine Complex in Aqueous Solution

Alkyl amines are able to form complexes with either crown ethers or cyclodextrins or cucurbit[6]uril. The same is known for polyamines such as spermidine and spermine. However, the simultaneous formation of such polyamines with crown ethers and cucurbit[6]uril has not been studied. The ability of polyamines such as spermidine and spermine to form mixed complexes with different ligands, e.g. crown ethers and cucurbit[6]uril has been studied in aqueous solution using pH-metric and calorimetric titrations. The thermodynamic data of reaction between crown ethers with spermidine, spermine and their cucurbit[6]uril complexes have been determined. The presence of cucurbit[6]uril on the polyamines has no important influence upon the reaction of these amines with crown ethers. The reactions between polyamines, cucurbit[6]uril and crown ethers are simple examples for the self organization of molecules due to specific interactions. Received in final form: 26 January 2005     

A Cucurbit[8]uril 2:2 Complex with a Negative pKa Shift

A viologen derivative carrying a benzimidazole group ( V-P-I ²⁺; viologen–phenylene–imidazole V-P-I ) can be dimerized in water using cucurbit[8]uril (CB[8]) in the form of a 2:2 complex resulting in a negative shift of the guest pK_a, by more than 1 pH unit, contrasting with the positive pK_a shift usually observed for CB-based complexes. Whereas 2:2 complex protonation is unclear by NMR, silver cations have been used for probing the accessibility of the imidazole groups of the 2:2 complexes. The protonation capacity of the buried imidazole groups is reduced, suggesting that CB[8] could trigger proton release upon 2:2 complex formation. The addition of CB[8] to a solution containing V-P- I ³⁺ indeed released protons as monitored by pH-metry and visualized by a coloured indicator. This property was used to induce a host/guest swapping, accompanied by a proton transfer, between V-P-I ³⁺ ⋅ CB[7] and a CB[8] complex of 1-methyl-4-(4-pyridyl)pyridinium. The origin of this negative pK_a shift is proposed to stand in an ideal charge state, and in the position of the two pH-responsive fragments inside the two CB[8] which, alike residues engulfed in proteins, favour the deprotonated form of the guest molecules. Such proton release triggered by a recognition event is reminiscent of several biological processes and may open new avenues toward bioinspired enzyme mimics catalyzing proton transfer or chemical reactions.     

Cucurbit[7]uril Complexation of Near-Infrared Fluorescent Azobenzene-Cyanine Conjugates

Two new azobenzene heptamethine cyanine conjugates exist as dispersed monomeric molecules in methanol solution and exhibit near-infrared (NIR) cyanine absorption and fluorescence. Both conjugates form non-emissive cyanine H-aggregates in water, but the addition of cucurbit[7]uril (CB7) induces dye deaggregation and a large increase in cyanine NIR fluorescence emission intensity. CB7 encapsulates the protonated azonium tautomer of the 4-(N,N-dimethylamino)azobenzene component of each azobenzene–cyanine conjugate and produces a distinctive new absorption band at 534 nm. The complex is quite hydrophilic, which suggests that CB7 can be used as a supramolecular additive to solubilize this new family of NIR azobenzene–cyanine conjugates for future biomedical applications. Since many azobenzene compounds are themselves potential drug candidates or theranostic agents, it should be possible to formulate many of them as CB7 inclusion complexes with improved solubility, stability, and pharmaceutical profile.     

Sequential Formation of Heteroternary Cucurbit[10]uril (CB[10]) Complexes

The globular and monocationic guest molecule trimethyl-azaphosphatrane ( AZAP , a protonated Verkade superbase) was shown to form a host:guest 1 : 1 complex with the cucurbit[10]uril (CB[10]) macrocycle in water. Molecular dynamics calculations showed that CB[10] adopts an 8-shape with AZAP occupying the majority of the internal space, CB[10] contracting around AZAP and leaving a significant part of the cavity unoccupied. This residual space was used to co-include planar and monocationic co-guest ( CG ) molecules, affording heteroternary CB[10]⋅ AZAP ⋅ CG complexes potentially opening new perspectives in supramolecular chemistry.     

Cucurbit[6]uril dimer induces supramolecular polymerisation of a cationic polyethylene glycol derivative

Recently, we reported the synthesis of two new dimeric cucurbit[6]uril compounds (1 and 2). Herein, we describe the ability of compound 1 to undergo supramolecular polymerisation when combined with polyethylene glycol (PEG) derived divalent guests 3_MW (MW = 300, 1000, 3350, 10,000). Interestingly, we find that the shorter PEG-derived guests 3₃₀₀, 3₁₀₀₀ and 3₃₃₅₀ lead to low degrees of oligomerisation based on the diffusion coefficients obtained by diffusion-ordered spectroscopy measurements. In contrast, when 1 was combined with the longest PEG-derived guest 3_10,000, we observed the formation of a supramolecular polymer with a degree of oligomerisation of at least 36. We observe that the supramolecular polymer 1_n·(3_10,000)n undergoes a slow decrease in its degree of oligomerisation to 22 over the course of 14 days controlled by the slow dissociation rate constant of the hexanediammonium unit of 3_10,000 from the CB[6]-sized cavity of 1.     

Selective Release of Doxorubicin from Cucurbit[8]uril Stabilized Gold Supra-Pyramid Host at pH of Small Intestine

Gold supra-pyramid structures were obtained by the addition of acidic solution of cucurbit[8]uril (CB[8]) to an aqueous solution of citrate stabilized gold nanoparticles (AuNP). The reaction resulted in the precipitation of supra-pyramid from the solution after just 1 min of shaking. Microscopic images confirmed formation of the supra-pyramid. The stepwise structural transformation towards the supra-pyramid was examined with variable concentrations of CB[8] to AuNP solution. Anionic counter parts of these acids (Br⁻, NO₃⁻, SO₄²⁻ and Cl⁻) controlled the size of the synthesized supra-pyramids. These supra-pyramid hosts showed uptake of three anticancer drugs: oral drugs etoposide, prednisolone and intravenous drug doxorubicin. Releases of drugs from these hosts were emulated at acidic stomach pH, basic small intestinal pH and in the presence of human serum albumin (HSA). The specific release of doxorubicin was confirmed at small intestinal pH 7.4. Poor release of drugs in presence of CB[8] specific guest 1-adamantanamine confirmed the role of the supra-pyramid as the exclusive host. The release of doxorubicin from the supra-pyramid at pH 7.4 was confirmed by fluorescence microscopic imaging with prostate cancer DU-145 cell line.     

Surface‐Bound Cucurbit[8]uril Catenanes on Magnetic Nanoparticles Exhibiting Molecular Recognition

We demonstrate the preparation of surface‐bound cucurbit[8]uril (CB[8]) catenanes on silica nanoparticles (NPs), where CB[8] was employed as a tethered supramolecular “handcuff” to selectively capture target guest molecules. In this catenane, CB[8] was threaded onto a methyl viologen (MV²⁺) axle and immobilized onto silica NPs. The formation of CB[8] catenanes on NPs were confirmed by UV/Vis titration experiments and lithographic characterization, demonstrating a high density of CB[8] on the silica NPs surface, 0.56 nm^?2. This CB[8] catenane system exhibits specific molecular recognition towards certain aromatic molecules such as perylene bis(diimide), naphthol and aromatic amino acids, and thus it can act as a nanoscale molecular receptor for target guests. Furthermore, we also demonstrate its use as an efficient and recyclable nano‐platform for peptide separation. By embedding magnetic NPs inside silica NPs, separation could be achieved by simply applying an external magnetic field. Moreover, the peptides captured by the catenanes could be released by reversible single‐electron reduction of MV²⁺. The entire process demonstrated high recoverability.     

High Affinity Recognition of a Selected Amino Acid Epitope within a Protein by Cucurbit[8]uril Complexation

Supramolecular interactions between the host cucurbit[8]uril (CB[8]) and amino acids have been widely interrogated, but recognition of specific motifs within a protein domain have never been reported. A phage display approach was herein used to select motifs with the highest binding affinity for the heteroternary complex with methyl viologen and CB[8] (MV?CB[8]) within a vast pool of cyclic peptide sequences. From the selected motifs, an epitope consisting of three amino acid was extrapolated and incorporated into a solvent‐exposed loop of a protein domain; the protein exhibited micromolar binding affinity for the MV?CB[8] complex, matching that of the cyclic peptide. By achieving selective CB[8]‐mediated conjugation of a small molecule to a recombinant protein scaffold we pave the way to biomedical applications of this simple ternary system.     

pH-Triggered dethreading-rethreading and switching of cucurbit[6]uril on bistable [3]pseudorotaxanes and [3]rotaxanes

A series of water-soluble [3]rotaxanes-(n+2) and [3]pseudorotaxanes-(n+2) with short (propyl, n=1) and long (dodecyl, n=10) aliphatic spacers have been prepared in high yields by a 1,3-dipolar cycloaddition reaction catalyzed by cucurbit[6]uril (CB6). The pH-triggered dethreading and rethreading of CB6 on these pseudorotaxanes was monitored by 1H NMR spectroscopy. A previously reported [3]rotaxane-12 that is known to behave as a bistable molecular switch has two recognition sites for CB6, that is, the diaminotriazole moieties and the dodecyl spacer. By changing the pH of the system, it is possible to observe more than one state in the shuttling process. At low pH values both CB6 units are located on the diaminotriazole moieties owing to an ion-dipole interaction, whereas at high pH values both of the CB6 units are located on the hydrophobic dodecyl spacer. Surprisingly, the CB6 units shuttle back to their initial state very slowly after reprotonation of the axle. Even after eighteen days at room temperature, only about 50 % of the CB6 units had relocated back onto the diaminotriazole moieties. The rate constants for the shuttling processes were measured as a function of temperature over the range from 313 to 333 K and the activation parameters (enthalpy, entropy, and free energy) were calculated by using the Eyring equation. The results indicate that this [3]rotaxane behaves as a kinetically controlled molecular switch. The switching properties of [3]rotaxane-3 have also been studied. However, even under extreme pH conditions this rotaxane has not shown any switching action, which confirms that the propyl spacer is too short to accommodate CB6 units.     

八元瓜环与抗癌药物甲氨蝶呤的超分子相互作用

利用紫外吸收光谱法考察了八元瓜环(Q[8])与抗癌药物甲氨蝶呤(MTX)的相互作用;同时研究了甲氨蝶呤的不同质子化形式及体系温度对相互作用的影响;计算了相应的稳定常数及热力学参数等.实验结果表明,Q[8]与MTX作用比为1∶1,包结平衡常数为(7.64±1.52)×104 L/mol;龄前热力学参数测定结果表明,上述主...     

Acyclic cucurbit[n]uril type receptors: secondary versus tertiary amide arms

ABSTRACT

Two acyclic CB[n]-type hosts (1 and 2) which possess four 2° or 3° amide arms are reported. Host 2 has four 3° amide arms that exist as a mixture of E- and Z-isomers. ¹H NMR was used to qualitatively investigate the binding properties of 1 and 2 which indicates they retain the essential binding features of macrocyclic CB[n] hosts. We measured the K_a values of 1 and 2 toward guests 6–14 by ITC. Neutral hosts 1 and 2 bind less tightly than tetraanionic hosts M1, ACB1, and ACB2. We attribute the lower K_a values to the absence of secondary ion-ion electrostatic interactions for host?guest complexes of 1 and 2. The secondary amide functionality on 1 decreases affinity by the formation of intramolecular NH???O=C H-bonds. Tertiary amide host 2 binds even more weakly than 1 due to backfolding of the amide N-CH₃-groups of 2 into its own cavity.     

Supramolecular Recognition of Amino Acids by Twisted Cucurbit[14]uril

Binding interactions between twisted cucurbit[14]uril (tQ[14]) and twenty standard amino acids (AAs) have been investigated by NMR spectroscopy and isothermal titration calorimetry (ITC) in aqueous HCl solutions and in DMSO. The results showed that tQ[14] displays clear binding affinity for AAs with a positively charged side chain or containing an aromatic ring, but weaker binding affinity for AAs with hydrophobic or polar side chains, with the binding mode depending on the type of side chain present in the AAs.     

Encapsulation-Controlled Photoisomerization of a Styryl Derivative: Stereoselective Formation of the Anti Z-Isomer in the Cucurbit[7]uril Cavity

The photochemical isomerization of a styrylpyridinium dye ( SP ) bearing an unsymmetrically attached benzo-15-crown-5 ether has been studied in aqueous solution in the absence and presence of cucurbit[7]uril (CB[7]). The detailed analysis of the UV/Vis and NMR spectra showes that the isomeric composition of the photostationary mixtures of SP can be modulated by the host-guest complexation with CB[7]. It was found that steric hindrance caused by encapsulation of SP in the host cavity induces the exclusive formation of the anti conformer of Z- SP in contrast with the mixture of both anti and syn conformers obtained during photoisomerization of the dye without CB[7]. Remarkably, the displacement of anti Z- SP from CB[7] does not lead to the transformation of the anti Z-isomer into the syn Z-isomer pointing out the conformational memory of the system. The results provide an interesting example of the supramolecular stereorecognition by the achiral CB[7] host.     

嘌呤衍生物在葫芦［6］脲单轮烷键合固定相上的色谱行为

在反相和正相色谱模式下,研究了几种嘌呤衍生物在葫芦［6］脲单轮烷键合硅胶固定相上的高效液相色谱行为,并在反相模式下与ODS固定相进行了比较,考察了流动相中甲醇含量、流动相pH值和离子强度对嘌呤化合物保留的影响。研究结果表明:在反相模式下,嘌呤化合物与葫芦［6］脲单轮烷键合相之间存在多种相互作用,除疏水作用外,分离过程中还存在与ODS不同的色谱分离机制。在正相条件下,多作用力的色谱分离机制同样存在。葫芦［6］脲单轮烷键合相与溶质之间存在疏水、氢键、π-π和偶极-偶极等多种作用力,协同作用提高了固定相对嘌呤化合物的分离选择性。     

V型菁染料与葫芦脲[8]的包结研究

近年来葫芦脲因为其在超分子化学领域有着巨大的潜在应用价值而备受研究者关注。本文描述了一种两臂均为葫芦脲潜在包结点的V型菁染料分子DP与葫芦脲[8]之间的包结现象,通过一维、二维核磁共振氢谱、质谱和紫外可见吸收光谱等手段对包结物进行了研究,结果表明DP与葫芦脲[8]形成1∶1包结物,且其两臂均被葫芦脲[8]所环绕,相关包结常数为(5.4±0.4)×105。