Fabrication of viable cyborg cells with cyclodextrin functionality

We describe two alternative methods for surface functionalisation of Saccharomyces cerevisiae cells with cyclodextrin molecules without affecting the cell viability. The first strategy involved using epichlorohydrin as a cross-linking agent which binds covalently the cyclodextrin to the glycoproteins on the cell wall. The second strategy of interfacing of the cells with CD involved polyelectrolyte mediated deposition of cyclodextrin sulphate on the cell surface. We used the formation of host-guest inclusion complexes of a dye with the grafted cyclodextrins to estimate the average number of CD molecules grafted per cell which can reach up to hundreds of millions of CD molecules. This indicates more than one monolayer of CDs on the cell surface within the surface layer surrounding the yeast cell membrane. Fluorescein diacetate was used to check the viability of the cells after functionalisation. Living cells functionalised with CDs may find many potential applications as they can be loaded with drugs, immunosuppressants and other molecules forming inclusion complexes with their cyclodextrin interface. Therefore, we foresee such cells being used as novel selective biosorbents in polluted waters, whole cell biosensors, drug delivery, cell therapy and cell implant applications

Delivery of the vitamin E compound tocotrienol to cancer cells

Tocotrienol, a member of the vitamin E family of compounds, is currently receiving increased attention because of its highly promising anti-cancer effects. However, its potential in cancer therapy is limited by its poor bioavailability and its inability to specifically reach tumors at therapeutic concentrations after intravenous administration. In order to remediate to these problems, various delivery strategies have been proposed, such as the inclusion of tocotrienol in γ-cyclodextrins, prodrugs and emulsions, entrapment in lipid nanoparticles and vesicles. Among these approaches, we demonstrated that the entrapment of tocotrienol within vesicles bearing transferrin, whose receptors are overexpressed on numerous cancer cells, significantly improved the uptake by cancer cells overexpressing transferrin receptors. Consequently, the intravenous administration of tocotrienol entrapped in transferrin-bearing vesicles led to tumor regression and even complete tumor suppression in some cases in a murine tumor model, as well as improvement of animal survival. Transferrin-bearing vesicles are therefore highly promising for the delivery of tocotrienol to cancer cells in vitro and in vivo and should be further investigated to optimize the anti-cancer therapeutic effect of tocotrienol

Engineering of Cyclodextrin Product Specificity and pH Optima of the Thermostable Cyclodextrin Glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1

The product specificity and pH optimum of the thermostable cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacterium thermosulfurigenes EM1 was engineered using a combination of x-ray crystallography and site-directed mutagenesis. Previously, a crystal soaking experiment with the Bacillus circulans strain 251 β-CGTase had revealed a maltononaose inhibitor bound to the enzyme in an extended conformation. An identical experiment with the CGTase from T. thermosulfurigenes EM1 resulted in a 2.6-Å resolution x-ray structure of a complex with a maltohexaose inhibitor, bound in a different conformation. We hypothesize that the new maltohexaose conformation is related to the enhanced α-cyclodextrin production of the CGTase. The detailed structural information subsequently allowed engineering of the cyclodextrin product specificity of the CGTase from T. thermosulfurigenes EM1 by site-directed mutagenesis. Mutation D371R was aimed at hindering the maltohexaose conformation and resulted in enhanced production of larger size cyclodextrins (β- and γ-CD). Mutation D197H was aimed at stabilization of the new maltohexaose conformation and resulted in increased production of α-CD. Glu258 is involved in catalysis in CGTases as well as α-amylases, and is the proton donor in the first step of the cyclization reaction. Amino acids close to Glu258 in the CGTase from T. thermosulfurigenes EM1 were changed. Phe284 was replaced by Lys and Asn327 by Asp. The mutants showed changes in both the high and low pH slopes of the optimum curve for cyclization and hydrolysis when compared with the wild-type enzyme. This suggests that the pH optimum curve of CGTase is determined only by residue Glu258.

Melarsoprol cyclodextrin inclusion complexes as promising oral candidates for the treatment of human African trypanosomiasis

Human African trypanosomiasis (HAT), or sleeping sickness, results from infection with the protozoan parasites <i>Trypanosoma brucei</i> (<i>T.b.</i>) <i>gambiense</i> or <i>T.b.rhodesiense</i> and is invariably fatal if untreated. There are 60 million people at risk from the disease throughout sub-Saharan Africa. The infection progresses from the haemolymphatic stage where parasites invade the blood, lymphatics and peripheral organs, to the late encephalitic stage where they enter the central nervous system (CNS) to cause serious neurological disease. The trivalent arsenical drug melarsoprol (Arsobal) is the only currently available treatment for CNS-stage <i>T.b.rhodesiense</i> infection. However, it must be administered intravenously due to the presence of propylene glycol solvent and is associated with numerous adverse reactions. A severe post-treatment reactive encephalopathy occurs in about 10% of treated patients, half of whom die. Thus melarsoprol kills 5% of all patients receiving it. Cyclodextrins have been used to improve the solubility and reduce the toxicity of a wide variety of drugs. We therefore investigated two melarsoprol cyclodextrin inclusion complexes; melarsoprol hydroxypropyl-͎-cyclodextrin and melarsoprol randomly-methylated-β-cyclodextrin. We found that these compounds retain trypanocidal properties <i>in vitro</i> and cure CNS-stage murine infections when delivered orally, once per day for 7-days, at a dosage of 0.05 mmol/kg. No overt signs of toxicity were detected. Parasite load within the brain was rapidly reduced following treatment onset and magnetic resonance imaging showed restoration of normal blood-brain barrier integrity on completion of chemotherapy. These findings strongly suggest that complexed melarsoprol could be employed as an oral treatment for CNS-stage HAT, delivering considerable improvements over current parenteral chemotherapy

Complexation between oleanolic and maslinic acids with native and modified cyclodextrins

Oleanolic (OA) and maslinic (MA) acids are two natural triterpenoids with a wide range of beneficial effects for human health. However, their low solubility and permeability make their application in the food or industry difficult. The complexation of OA and MA with alpha- beta-, gamma-, HP-alpha-, HP-beta- and HP-gamma-CDs under different pH and temperature conditions has been studied. Neither alpha- nor HP-alpha-CDs formed inclusion complexes, while beta-, HP-beta- and HP-gamma-CDs provided AL type and gamma-CDs BS phase solubility diagrams. Complexation was shown to be more stable in the case of MA but complexation efficiency was greater for OA. Increasing the pH and temperature of the complexation media tended to improve the complexation process with triterpenic acids.Agencia de Ciencia y Tecnología de la Región de Murcia under the project PFEseneca/06/10Ciencias de la Alimentació

A hopeful therapy for Niemann-Pick C diseases

Not abstract availabl

Vibrational properties of inclusion complexes: the case of indomethacin-cyclodextrin

Vibrational properties of inclusion complexes with cyclodextrins are studied by means of Raman spectroscopy and numerical simulation. In particular, Raman spectra of the non-steroidal, anti-inflammatory drug indomethacin undergo notable changes in the energy range between 1600 and 1700 cm$^{-1}$ when inclusion complexes with cyclodextrins are formed. By using both \emph{ab initio} quantum chemical calculations and molecular dynamics, we studied how to relate such changes to the geometry of the inclusion process, disentangling single-molecule effects, from changes in the solid state structure or dimerization processes.Comment: 14 file figure

The influence of the preparation methods on the inclusion of model drugs in a β-cyclodextrin cavity

NOTICE: this is the author’s version of a work that was accepted for publication in European Journal of Pharmaceutics and Biopharmaceutics. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Eur J Pharm Biopharm. 2009 Feb;71(2):377-386. Epub 2008 Oct 17.The work aims to prove the complexation of two model drugs (ibuprofen, IB and indomethacin, IN) by bcyclodextrin (bCD), and the effect of water in such a process, and makes a comparison of their complexation yields. Two methods were considered: kneading of a binary mixture of the drug, bCD, and inclusion of either IB or IN in aqueous solutions of bCD. In the latter method water was removed by air stream, spray-drying and freeze-drying. To prove the formation of complexes in final products, optical microscopy, UV spectroscopy, IR spectroscopy, DSC, X-ray and NMR were considered. Each powder was added to an acidic solution (pH = 2) to quantify the concentration of the drug inside bCD cavity. Other media (pH = 5 and 7) were used to prove the existence of drug not complexed in each powder, as the drugs solubility increases with the pH. It was observed that complexation occurred in all powders, and that the fraction of drug inside the bCD did not depend neither on the method of complexation nor on the processes of drying considered

Effects of Cyclodextrin Type on Vitamin C, Antioxidant Activity, and Sensory Attributes of a Mandarin Juice Enriched with Pomegranate and Goji Berries

The effects of the addition of cyclodextrins (CDs), β-CD, or HP-β-CD (1%), on the protection of antioxidant compounds of mandarin juices enriched with pomegranate extract and goji berries juice, was studied. Juices were prepared and after their thermal treatment (98 ◦C, 30 s) they were stored at 4 ◦C during 75 d. Vitamin C content, CIE L∗a∗b∗ color, antioxidant capacity, retinol equivalents, and sensory properties were studied. Losses on vitamin C were higher (6%) for juices with β-CD than juices with HP-β-CD. Retinol equivalents degradation was lower (3.4%) in juices with HP-β-cyclodextrins than in those treated with β-CD. Lower losses were observed for the instrumental and sensory color intensity in juices with HP-β-CD addition. Finally, the antioxidant capacity was also higher in juices treated with HP-β-CD. Finally, the overall sensory quality of juices with HP-β-CD was the best one after 30 d of cold storage. Even though β-CD addition did not cause any improvement compared with control juice (without CD addition), the benefits of adding HP-β-CD to this particular juice were shown in almost all parameters under study.Ciencias de la Alimentació