Development of a novel injectable drug delivery system for subconjunctival glaucoma treatment.

In this study we present the development of an injectable polymeric drug delivery system for subconjunctival treatment of primary open angle glaucoma. The system consists of hyaluronic acid sodium salt (HA), which is commonly used in ophthalmology in anterior segment surgery, and an isocyanate-functionalized 1,2-ethylene glycol bis(dilactic acid) (ELA-NCO). The polymer mixtures with different ratios of HA to ELA-NCO (1/1, 1/4, and 1/10 (v/v)) were investigated for biocompatibility, degradation behavior and applicability as a sustained release system. For the latter, the lipophilic latanoprost ester pro-drug (LA) was incorporated into the HA/ELA-NCO system. In vitro, a sustained LA release over a period of about 60days was achieved. In cell culture experiments, the HA/ELA-NCO (1/1, (v/v)) system was proven to be biocompatible for human and rabbit Tenon's fibroblasts. Examination of in vitro degradation behavior revealed a total mass loss of more than 60% during the observation period of 26weeks. In vivo, LA was continuously released for 152days into rabbit aqueous humor and serum. Histological investigations revealed a marked leuko-lymphocytic infiltration soon after subconjunctival injection. Thereafter, the initial tissue reaction declined concomitantly with a continuous degradation of the polymer, which was completed after 10months. Our study demonstrates the suitability of the polymer resulting from the reaction of HA with ELA-NCO as an injectable local drug delivery system for glaucoma therapy, combining biocompatibility and biodegradability with prolonged drug release.

In vitro degradation and drug release of a biodegradable tissue adhesive based on functionalized 1,2-ethylene glycol bis(dilactic acid) and chitosan.

Biodegradability and adhesive-associated local drug release are important aspects of research in tissue adhesive development. Therefore, this study focuses on investigating the in vitro degradation and drug release of a tissue adhesive consisting of hexamethylene diisocyanate functionalized 1,2-ethylene glycol bis(dilactic acid) and chitosan chloride. To prevent infections, ciprofloxacin hydrochloride (CPX·HCl) was incorporated into the adhesive. The influence of CPX·HCl on the adhesive reaction and adhesive strength was analyzed by FTIR-ATR-spectroscopy and tensile tests. The CPX·HCl release was investigated by HPLC. The degradation-induced changes at 37 °C were evaluated by gravimetric/morphological analyzes and micro-computer tomography. The antibiotic potential of the CPX·HCl loaded adhesive was determined by agar diffusion tests. The degradation tests revealed a mass loss of about 78 % after 52 weeks. The adhesive reaction velocity and tensile strength were not influenced by CPX·HCl. Using a 2 mg/g CPX·HCl loaded adhesive an inhibition of all tested bacteria was observed.

The cyanogenic syndrome in rubber tree Hevea brasiliensis: tissue-damage-dependent activation of linamarase and hydroxynitrile lyase accelerates hydrogen cyanide release.

BACKGROUND AND AIMS: The release of hydrogen cyanide (HCN) from injured plant tissue affects multiple ecological interactions. Plant-derived HCN can act as a defence against herbivores and also plays an important role in plant-pathogen interactions. Crucial for activity as a feeding deterrent is the amount of HCN generated per unit time, referred to as cyanogenic capacity (HCNc). Strong intraspecific variation in HCNc has been observed among cyanogenic plants. This variation, in addition to genotypic variability (e.g. in Trifolium repens), can result from modifications in the expression level of the enzymes involved in either cyanogenic precursor formation or HCN release (as seen in Sorghum bicolor and Phaseolus lunatus). Thus, a modification or modulation of HCNc in reaction to the environment can only be achieved from one to the next generation when under genetic control and within days or hours when transcriptional regulations are involved. In the present study, it is shown that in rubber tree (Hevea brasiliensis) HCNc is modulated by post-translational activity regulation of the key enzymes for cyanide release. METHODS: Linamarase (LIN) and hydroxynitrile lyase (HNL) activity was determined by colorimetric assays utilizing dissociation of the substrates p-nitrophenyl-ß-d-glucopyranoside and acetone cyanohydrin, respectively. KEY RESULTS: In rubber tree leaves, LIN and HNL show up to ten-fold increased activity in response to tissue damage. This enzyme activation occurs within seconds and results in accelerated HCN formation. It is restricted to the damaged leaf area and depends on the severity of tissue damage. CONCLUSIONS: LIN and HNL activation (in contrast to genetic and transcriptional regulations) allows an immediate, local and damage type-dependent modulation of the cyanogenic response. Accordingly, this post-translational activation plays a decisive role in the defence of H. brasiliensis against herbivores as well as pathogens and may allow more flexible reactions in response to these different antagonists.

Ionic liquids containing the triply negatively charged tricyanomelaminate anion and a B(C6F5)3 adduct anion.

Room-temperature ionic liquids containing the triply charged tricyanomelaminate (tcmel) ion [C(3)N(6)(CN)(3)](3-) were synthesized. The 1-methyl-3-methylimidazolium (MMIm), 1-ethyl-3-methylimidazolium (EMIm), and 1-butyl-3-methylimidazolium (BMIm) salts of the tricyanomelaminate ion have glass transition temperatures (-6, -20, and -30 °C) similar to those found for the analogous monomeric dicyanoamide salts. They are thermally stable up to over 200 °C and dissolve in polar organic solvents. Addition of B(C(6)F(5))(3) to M(3) [tcmel] (M=Na, MMIm, EMIm, BMIm) yields salts containing the very voluminous adduct ion [C(3)N(6) {CN·B(C(6)F(5))(3)}(3)](3-) (tcmel_3B). The solid-state structure of [MMIm](3-) [tcmel] shows only long cation···anion contacts but in large number, while the solid-state structure of [Na(THF)(3)](3-) [tcmel_3B]·1.76 THF displays strong interactions of the sodium cation with the amido nitrogen atoms of the anion. Hence this adduct anion cannot be regarded as a weakly coordinating anion. A similar situation is found for the MMIm salt, [MMIm](3) [tcmel_3B]·2.66 CH(2)Cl(2), in which weak hydrogen bonds with the acidic proton of the MMIm ion are observed. On the basis of computations the energetics, structural trends, and charge transfer of adduct anion formation were studied.

Vaporization and formation enthalpies of 1-alkyl-3-methylimidazolium tricyanomethanides.

Thermochemical studies of the ionic liquids 1-ethyl-3-methylimidazolium tricyanomethanide [C(2)MIM][C(CN)(3)] and 1-butyl-3-methylimidazolium tricyanomethanide [C(4)MIM][C(CN)(3)] have been performed in this work. Vaporization enthalpies have been obtained using a recently developed quartz crystal microbalance (QCM) technique. The molar enthalpies of formation of these ionic liquids in the liquid state were measured by means of combustion calorimetry. A combination of the results obtained from QCM and combustion calorimetry lead to values of gaseous molar enthalpies of formation of [C(n)MIM][C(CN)(3)]. First-principles calculations of the enthalpies of formation in the gaseous phase for the ionic liquids [C(n)MIM][C(CN)(3)] have been performed using the CBS-QB3 and G3MP2 theory and have been compared with the experimental data. Furthermore, experimental results of enthalpies of formation of imidazolium-based ionic liquids with the cation [C(n)MIM] (where n = 2 and 4) and anions [N(CN)(2)], [NO(3)], and [C(CN)(3)] available in the literature have been collected and checked for consistency using a group additivity procedure. It has been found that the enthalpies of formation of these ionic liquids roughly obey group additivity rules.

Imidazolium-based ionic liquids. 1-methyl imidazolium nitrate: thermochemical measurements and ab initio calculations.

In this work data of the molar enthalpies of formation of the ionic liquid 1-methylimidazolium nitrate [H-MIM][NO3] was measured by means of combustion calorimetry. The molar enthalpy of fusion of [H-MIM][NO3] was measured using DSC. Experiments to vaporize the ionic liquid into vacuum or nitrogen stream in order to obtain vaporization enthalpy have been performed. Ab initio calculations of the enthalpy of formation in the gaseous phase have been performed for the ionic species using the G3MP2 theory. The combination of traditional combustion calorimertry with modern high-level ab initio calculations allow the determination of the molar enthalpy of vaporization of the ionic liquid under study. The ab initio calculations indicate that [H-MIM][NO3] is most probably separated into the neutral species methyl-imidazole and HNO3 in the gaseous phase at conditions of the vaporization experiments.

Synthesis, structure, and bonding of weakly coordinating anions based on CN adducts.

The addition of alkali or silver salts of dicyanoamide (dca), tricyanomethanide (tcm) and tetracyanoborate (tcb) to a solution of B(C(6)F(5))(3) in diethyl ether affords salts containing very voluminous B(C(6)F(5))(3) adduct anions of the type [E(CN)(n)(-)] x [B(C(6)F(5))(3)](n): E = N (dca_nb with n = 1, 2; b = B(C(6)F(5))(3)); E = C (tcm_nb with n = 1, 2, 3), and E = B (tcb_nb with n = 1, 2, 3, 4). Salts bearing these anions such as B[(CN) x B(C(6)F(5))(3)](4)(-) (= [B(CN)(4)(-)] x [B(C(6)F(5))(3)](4)), C[(CN) x B(C(6)F(5))(3)](3)(-) (= [C(CN)(3)(-)] x [B(C(6)F(5))(3)](3)), and N[(CN) x B(C(6)F(5))(3)](2)(-) (=[N(CN)(2)(-)] x [B(C(6)F(5))(3)](2)) can be prepared in good yields. They are thermally stable up to over 200 degrees C and dissolve in polar organic solvents. Depending on the stoichiometry mono-, di-, tri-, or tetraadduct formation is observed. The solid state structures of dca_2b, tcm_3b and tcb_4b salts show only long cation...anion contacts and thereby weak interactions, large anion volumes and only small distortions of the dca, tcm or tcb core enwrapped between B(C(6)F(5))(3) groups. That is why these anions can be regarded as weakly coordinating anions. On the basis of B3LYP/6-31+G(d) computations the energetics, structural trends and charge transfer of the adduct anion formation were studied. Since tcm_3b and tcb_4b are easily accessible and can also be prepared in large quantities, these anions may be utilized as a true alternative to other widely used weakly coordinating anions. Moreover, for both steric and electronic reasons it seems reasonable to expect that as counterions for cationic early transition metal catalysts such anions may show reduced ion pairing and hence increased catalytic activity.