Deformable liposomes containing alkylcarbonates of γ-cyclodextrins for dermal applications

Liposomes made with hydrogenated soya lecithin (HPC) mixed with dodecylcarbonate γ-cyclodextrin (C12CD) at 20:1, 10:1 and 5:1 w/w ratios were prepared by the solvent evaporation method. C12CD had emulsifying properties and the possibility of producing deformable liposomes, as topical delivery system of progesterone (PG), was evaluated. Liposome size, deformability and drug entrapment were determined and the interaction between C12CD and HPC was investigated using differential scanning calorimetry (DSC). The size and the amount of PG loaded in the liposomes depended on the lipid:C12CD ratio: the smallest liposomes were obtained using 20:1 ratio and the maximum drug entrapment at 5:1 ratio. DSC analysis suggested that C12CD interacted with liposomes disrupting and fluidizing the lipid bilayer. PG transepidermal permeation through intact pig skin and PG skin uptake from deformable liposomes were assessed and compared to the values obtained from aqueous suspension and conventional liposomes. The PG permeations were negligible for all systems, while skin uptake increased for liposomes containing C12CD. This was attributed to the deformability and to the increase in the drug entrapment efficiency of these liposomes. The use of C12CD in liposome formulations can improve PG topical therapy.     

Flow-injection chemiluminometric analysis of some steroids by their sensitizing effect on the bromate-sulphite reaction

The chemiluminescent oxidation of sulphite by bromate was investigated and compared with that by cerium(IV). The reaction is sensitized by various steroid hormones which can thus be determined in the ranges 0.50–20.0μg ml ¹ for cortisone; 0.50–5.00 μg ml^?1 for hydrocortisone and progesterone and 0.50–6.00 μg ml^?1 for testosterone and corticosterone.     

A comparative study of the adsorption equilibrium of progesterone by a carbon black and a commercial activated carbon

In this paper the adsorption process of a natural steroid hormone (progesterone) by a carbon black and a commercial activated carbon has been studied. The corresponding equilibrium isotherms have been analyzed according to a previously proposed model which establishes a kinetic law satisfactorily fitting the C versus t isotherms. The analysis of the experimental data points out the existence of two well-defined sections in the equilibrium isotherms. A general equation including these two processes has been proposed, the global adsorption process being fitted to such equation. From the values of the kinetic equilibrium constant so obtained, values of standard average adsorption enthalpy () and entropy () have been calculated. Finally, information related to variations of differential adsorption enthalpy () and entropy () with the surface coverage fraction (θ) was obtained by using the corresponding Clausius-Clapeyron equations.     

Norcholanic acids as substrates for recombinant 3β-hydroxysteroid dehydrogenase and progesterone 5β-reductase, enzymes of the 5β-cardenolide biosynthesis

The conversion of 23-nor-5,20(22)E-choladienic acid-3β-ol and other intermediates of the putative norcholanic acid pathway of cardenolide biosynthesis by recombinant 3β-hydroxysteroid dehydrogenase from Digitalis lanata in dehydrogenation and reduction reactions was investigated. 23-nor-4,20(22)E-choladienic acid-3-one was found to be a substrate of recombinant progesterone 5β-reductases from D. lanata and Arabidopsis thaliana. The role of various substrates in cardenolide biosynthesis is discussed.     

Photo-induced electron emission from 17beta-estradiol and progesterone and possible biological consequences

It was established for the first time, that the sexual hormones 17beta-estradiol (17betaE2) and progesterone (PRG) are able to emit electrons from their excited single state in water-ethanol mixtures. The yield of the "solvated electrons" (e(s)(-)) depends on the substrate concentration, the ratio of water-alcohol-mixtures and the temperature. The e(s)(-) yield obtained from 17betaE2 is by two orders of magnitude higher than this of PRG. The possible relationship of the resulting hormone transients from 17betaE2 leading via specific metabolites to breast cancer is discussed.     

A Dual-Label Time-Resolved Fluorescence Immunoassay for the Simultaneous Determination of β-Human Chorionic Gonadotropin and Progesterone

A dual-label time-resolved fluorescence immunoassay (TRFIA) for the simultaneous determination of β human chorionic gonadotropin and progesterone was developed for the early diagnosis of ectopic pregnancy. The performance of this assay was evaluated using clinical serum and then compared with standard procedures. The sensitivity for β human chorionic gonadotropin detection was 1 U/L (linear dynamic range, 0–8000 U/L), and the sensitivity for progesterone detection was 0.05 ng/mL (linear dynamic range, 0–50 ng/mL). High correlation coefficients (R²) were obtained between the reported immunoassay and standard methods (R² = 0.99 for β human chorionic gonadotropin, R² = 0.97 for progesterone). The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical analyses and is a suitable alternative to the single-labeled diagnostic methods.     

Nanoparticles derived from amphiphilic γ-cyclodextrins

Three alkylcarbonates of γ-cyclodextrin, i.e. hexyl, octyl and dodecylcarbonate, were synthesized and characterized, with the goal of formulating solid nanoparticles. The series of alkylcarbonates showed amphiphilic properties and were capable of forming micelles and nanoparticles. Blank and drug-loaded alkylcarbonate nanoparticles were prepared with each alkylcarbonate, using the solvent injection technique. Progesterone was chosen as model drug. The sizes of both unloaded and loaded nanoparticles were in the 80–200 nm range, with narrow size distribution and spherical shape, as shown by TEM analysis. The zeta potentials of unloaded nanoparticles were in the −20 to −24.0 mV range, and were slightly decreased in loaded nanoparticles. Drug-loading capacity was good; DSC analysis did not detect the progesterone melting peak, indicating the drug had interacted with the cyclodextrin alkylcarbonates. In vitro release kinetics of progesterone from the three types of nanoparticles were slow. These results indicate that γ-CD alkylcarbonate nanoparticles might be used as prolonged drug delivery system.     

Chemical structure characterization of polysaccharide from Osmunda japonica Thunb and its inhibitory activity on uterine fibroids

The active component polysaccharide of Osmunda japonica Thunb was extracted and its chemical structure was detected by FT-IR, 1D-NMR and 2D-NMR spectra, in order to have a positive preventive and therapeutic effect on uterine fibroids. The physicochemical properties of Osmunda japonica Thunb polysaccharides (OPP) were analyzed, and the results showed that OPP was heteropolysaccharides composed of six monosaccharides. Ara and Glc were the main monosaccharides of the polysaccharide, with a molecular weight of 252.54 ± 10.84 kDa. The main chain was connected by 1,3-linked Araf, which had a high degree of branching, and was connected to the branch at the O-2 and O-3 positions. The mice model of uterine fibroids was established by the estrogen load method. The coefficient of the uterus in the mice model increased significantly, irregular nodular processes were observed in the uterus, focal hyperplasia appeared in the smooth muscle layer, cell enlargement, disordered arrangement of smooth muscle cells, and endometrial hyperplasia was obvious. Furthermore, this work found that the serum estradiol (E2) and progesterone (P) concentrations in the model group were significantly higher than those in the normal group (P < 0.01), indicating successful modeling. OPP decreased serum E2 and P levels in a dose-dependent manner. Collectively, OPP had an obvious inhibitory effect on uterine fibroids in estrogen-loaded mice.     

孕血和药物中孕酮的单扫极谱法测定

孕酮２，４－二硝基苯腙在单扫描示波极谱仪上，以吡啶－氢氧化钠为底液（ｐＨ２．０）于－０．７４Ｖ（ｖｓ．ＳＣＥ）处可出现一灵敏的吸附催化波，孕酮浓度在１．０×１０＾－８～２．０×１０＾－６ｍｏｌ／Ｌ范围内与该波呈线性关系，检出限为５．０×１０＾－９ｍｏｌ／Ｌ方法用于测定孕血和药物中孕酮含量，获得满意的结果。     

Water soluble progesterone–hydroxypropyl-β-cyclodextrin complex for injectable formulations

The use of cyclodextrin to increase the water solubility of progesterone (P) was described by Pitha as a complex with β-cyclodextrin and derivates to obtain a water soluble formulation (Pitha, J.: US patent n. 4,727,064). Hydroxypropyl-β-cyclodextrin (HPBCD) has a high water solubility which allows the solubilization of high quantity of P. Considering a 1:2 guess/host complex stoichiometry it is possible to obtain up to 50 mg/ml of P concentration, which is a considerable dosage for drug development in the progesterone therapy. In our drug development the P/HPBCD complex in water showed the formation of a light precipitate during stability ICH conditions. A precipitate formation was described already by Choi (J. Korean Pharm. Sci. 31(3), 151, 2001) and also by Pitha (US patent n. 4,727,064) but the chemical structure was not elucidated. In our case the precipitate was purified and it turned out to contain progesterone and residual unmodified β-cyclodextrin. We have developed a production process in which the residual unreacted β-cyclodextrin is separated from the HPBCD by the formation of the insoluble inclusion complex (Zoppetti et al.: European Patent deposit n. 05108494.5). The resulting P/HPBCD contains up to 0.1% of residual β-cyclodextrin and does not produce precipitate during the stability study. The complex stoichiometry and the complex constant were calculated by the phase solubility study according to Higuchi and Connors (Adv. Anal. Chem. Instrum. 4, 117, 1965) and the presence of the inclusion complex was demonstrated by DSC, NMR, X-ray, FTIR. The formulation prepared at pilot scale as injectable form compared with the commercial oil formulation demonstrated a favourable kinetic in humans.