In vitro anticancer activity of Betulinic acid and derivatives thereof on equine melanoma cell lines from grey horses and in vivo safety assessment of the compound NVX-207 in two horses.

Betulinic acid, a pentacyclic triterpene, and its derivatives are promising compounds for cancer treatment in humans. Melanoma is not only a problem for humans but also for grey horses as they have a high potential of developing melanoma lesions coupled to the mutation causing their phenotype. Current chemotherapeutic treatment carries the risk of adverse health effects for the horse owner or the treating veterinarian by exposure to antineoplastic compounds. Most treatments have low prospects for systemic tumor regression. Thus, a new therapy is needed. In this in vitro study, Betulinic acid and its two derivatives B10 and NVX-207, both with an improved water solubility compared to Betulinic acid, were tested on two equine melanoma cell lines (MelDuWi and MellJess/HoMelZh) and human melanoma (A375) cell line. We could demonstrate that all three compounds especially NVX-207 show high cytotoxicity on both equine melanoma cell lines. The treatment with these compounds lead to externalization of phosphatidylserines on the cell membrane (AnnexinV-staining), DNA-fragmentation (cell cycle analysis) and activation of initiator and effector caspases (Caspase assays). Our results indicate that the apoptosis is induced in the equine melanoma cells by all three compounds. Furthermore, we succeed in encapsulating the most active compound NVX-207 in 2-Hydroxyprolyl-ß-cyclodextrine without a loss of its activity. This formulation can be used as a promising antitumor agent for treating grey horse melanoma. In a first tolerability evaluation in vivo the formulation was administered every one week for 19 consecutive weeks and well tolerated in two adult melanoma affected horses.

Assessment of solubilization characteristics of different surfactants for carvedilol phosphate as a function of pH.

The effect of surfactants on the solubility of a new phosphate salt of carvedilol was investigated at different biorelevent pH to evaluate their solubilization capacity. Solutions of different classes of surfactants viz., anionic-sodium dodecyl sulfate (SDS) and sodium taurocholate (STC), cationic-cetyltrimethylammonium bromide (CTAB) and non-ionic-Tween 80 (T80) were prepared in the concentration range of 5-35 mmol dm(-3) in buffer solutions of pH 1.2, 3.0, 4.5, 5.8, 6.8 and 7.2. The solubility data were used to calculate the solubilization characteristics viz. molar solubilization capacity, water micelle partition coefficient, free energy of solubilization and binding constant. Solubility enhancement in basic pH was in following order: CTAB>T80>SDS>STC. CTAB and T80 showed remarkable solubility enhancement in acidic pH as well. Among the anionic surfactants, solubility in acidic medium was retarded except at pH 1.2 in case of SDS. Cationic and non-ionic surfactants were found to be suitable for enhancing the solubility of CP which can be employed for maintaining the in vitro sink condition in the basic dissolution medium. While anionic surfactants showed solubility retardant behavior which may be exploited in increasing the drug entrapment efficiency of a colloidal drug delivery system formulated by emulsification technique.

Evaluation of dynamic polar molecular surface area as predictor of drug absorption: comparison with other computational and experimental predictors.

The relationship between various molecular descriptors and transport of drugs across the intestinal epithelium was evaluated. The monolayer permeability (Pc) of human intestinal Caco-2 cells to a series of nine beta-receptor-blocking agents was investigated in vitro. The dynamic polar molecular surface area (PSAd) of the compounds was calculated from all low-energy conformations identified in molecular mechanics calculations in vacuum and in simulated chloroform and water environments. For most of the investigated drugs, the effects of the different environments on PSAd were small. The exception was H 216/44, which is a large flexible compound containing several functional groups capable of hydrogen bonding (PSAd,chloroform = 70.8 A2 and PSAd,water = 116.6 A2). The relationship between Pc and PSAd was stronger than those between Pc and the calculated octanol/water distribution coefficients (log Dcalc) or the experimentally determined immobilized liposome chromatography (ILC) retention. Pc values for two new practolol analogues and H 216/44 were predicted from the structure-permeability relationships of a subset of the nine compounds and compared with experimental values. The Pc values of the two practolol analogues were predicted well from both PSAd calculations and ILC retention studies. The Pc value of H 216/44 was reasonably well-predicted only from the PSAd of conformations preferred in vacuum and in water. The other descriptors overestimated the Pc of H 216/44 100-500-fold.

Environmental assessment of the alkanolamines.

This review provides a summary of current information available on the environmental fate and aquatic toxicology of the alkanolamines. Because these materials are widely used, there is a need to understand their fate and effects in the environment. This assessment was confined to information regarding selected physical properties of the alkanolamines as well as their potential for degradation in the atmosphere, soil, surface water, and groundwater. In addition, their relevant aquatic toxicological information and bioconcentration potential were evaluated. In general, the alkanolamines have high water solubilities and low to moderate vapor pressures. Some are solids whereas others are liquids at room temperature. Aqueous solutions of the alkanolamines are basic, with the pKas decreasing with increased alkyl substitution. Predictions of the environmental distribution of these compounds, based on a unit world model of Mackay and Paterson, suggested that alkanolamines would partition primarily into the aqueous compartment at equilibrium, with the remainder distributed to the atmosphere. Only a very small fraction of these materials is expected to sorb to soil or sediments. However, adsorption mechanisms other than partitioning into the soil organic layer were not considered in this model. Since polar compounds may sorb to soil by alternate mechanisms, this model may underestimate the true adsorption potential and subsequent environmental distribution of the alkanolamines. Future work with these compounds should focus on other types of adsorption mechanisms that could impact the environmental distribution of the alkanolamines. Although only small amount of the alkanolamines are expected to partition to the atmosphere, they are expected to be removed by reactions with photochemically generated hydroxyl radicals. They may also be removed from the atmosphere by precipitation, due to their high water solubility. Because of the relatively low levels expected to be present in the atmosphere and the relatively short half-lives, the alkanolamines are not expected to adversely impact air quality. Alkanolamines have also been shown to be highly susceptible to biodegradation and are not expected to persist in the environment. Results from numerous studies have shown that these materials undergo rapid biodegradation in soil, surface waters, and wastewater treatment plants. Degradation rates for these compounds may vary, with half-lives routinely in the range of 1 d to 2 wk, depending on the length of acclimation period and other environmental factors. The relatively low bioconcentration factor (BCF) values reported for the alkanolamines indicate that they would not be expected to bioconcentrate in aquatic organisms. Available data on the toxicity of the alkanolamines to aquatic organisms suggest low toxicity to the majority of the species studied. Based on the facts that alkanolamines exhibit low aquatic toxicity, are shown to biodegrade in a wide range of environments, and exhibit no tendency to bioaccumulate, the routine manufacturing, use, and disposal of these materials are not expected to adversely impact the environment. With increased emphasis by consumers and regulatory agencies for industry to develop products that are "environmentally friendly," these properties of the alkanolamines make them an attractive choice for a wide range of applications.