Dissolution enhancement of active pharmaceutical ingredients by therapeutic deep eutectic systems

A therapeutic deep eutectic system (THEDES) is here defined as a deep eutectic solvent (DES) having an active pharmaceutical ingredient (API) as one of the components. In this work, THEDESs are proposed as enhanced transporters and delivery vehicles for bioactive molecules. THEDESs based on choline chloride (ChCl) or menthol conjugated with three different APIs, namely acetylsalicylic acid (AA), benzoic acid (BA) and phenylacetic acid (PA), were synthesized and characterized for thermal behaviour, structural features, dissolution rate and antibacterial activity. Differential scanning calorimetry and polarized optical microscopy showed that ChCl:PA (1:1), ChCl:AA (1:1), menthol:AA (3:1), menthol:BA (3:1), menthol:PA (2:1) and menthol:PA (3:1) were liquid at room temperature. Dissolution studies in PBS led to increased dissolution rates for the APIs when in the form of THEDES, compared to the API alone. The increase in dissolution rate was particularly noticeable for menthol-based THEDES. Antibacterial activity was assessed using both Gram-positive and Gram-negative model organisms. The results show that all the THEDESs retain the antibacterial activity of the API. Overall, our results highlight the great potential of THEDES as dissolution enhancers in the development of novel and more effective drug delivery systems.

Development of folate-targeted liposomes for rheumatoid arthritis therapy

Tese de Doutoramento em Biologia Molecular e Ambiental (área de especialização em Biologia Celular e Saúde).

Improved burn wound healing using a bioactive peptide

[Excerpt] Antimicrobial peptides (AMPs) are good candidates to treat burn wounds, a major cause of morbidity, impaired life quality and resources consumption in developed countries. We took advantage of a commercially available hydrogel, Carbopol, a vehicle for topical administration that maintains a moist environment within the wound site. We hypothesized that the incorporation of LLKKK18 conjugated to dextrin would improve the healing process in rat burns. Whereas the hydrogel improves healing, LLKKK18 released from the dextrin conjugates further accelerates wound closure, and simultaneously improving the quality of healing. Indeed, the release of LLKKK18 reduces oxidative stress and inflammation (low neutrophil and macrophage infiltration and pro-inflammatory cytokines levels). Importantly, it induced a faster resolution of the inflammatory stage through early M2 macrophage recruitment. In addition, LLKKK18 stimulates angiogenesis (increased VEGF and microvessel development in vivo), potentially contributing to more effective transport of nutrients and cytokines. Moreover, collagen staining evaluated by Masson’s Trichrome was visually much more intense after treatment with LLKKK18, suggesting higher collagen deposition. (...)

Presence of starch enhances in vitro biodegradation and biocompatibility of a gentamicin delivery formulation

The effect of α-amylase degradation on the release of gentamicin from starch-conjugated chitosan microparticles was investigated up to 60 days. Scanning electron microscopic observations showed an increase in the porosity and surface roughness of the microparticles as well as reduced diameters. This was confirmed by 67% weight loss of the microparticles in the presence of α-amylase. Over time, a highly porous matrix was obtained leading to increased permeability and increased water uptake with possible diffusion of gentamicin. Indeed, a faster release of gentamicin was observed with α-amylase. Starch-conjugated chitosan particles are non-toxic and highly biocompatible for an osteoblast (SaOs-2) and fibroblast (L929) cell line as well as adipose-derived stem cells. When differently produced starch-conjugated chitosan particles were tested, their cytotoxic effect on SaOs-2 cells was found to be dependent on the crosslinking agent and on the amount of starch used.

Reactivity studies on chromene derivatives

Dissertação de mestrado em Medicinal Chemistry

Comparison of the effect of two HMG CoA reductase inhibitors on LDL susceptibility to oxidation

OBJECTIVE: To study the differences between fluvastatin and pravastatin regarding LDL susceptibility to oxidation, plasma levels of total cholesterol (TC), HDL-C, LDL-C and triglycerides (TG) in hypercholesterolemic patients with established coronary heart disease (CHD). METHODS: A double-blind randomized parallel study was conducted that included 41 hypercholesterolemic outpatients with CHD treated at the Instituto de Cardiologia do Rio Grande do Sul. The inclusion criteria were LDL-C above 100 mg/dL and triglycerides below 400 mg/dL based on 2 measures. After 4 weeks on a low cholesterol diet, those patients that fullfilled the inclusion criteria were randomized into 2 groups: the fluvastatin group (fluvastatin 40 mg/day) and the pravastatin group (pravastatin 20 mg/day), for 24 weeks of treatment. LDL susceptibility to oxidation was analyzed with copper-induced production of conjugated dienes (Cu2+) and water-soluble free radical initiator azo-bis (2'-2'amidinopropanil) HCl (AAPH). Spectroscopy nuclear magnetic resonance was used for determination of lipids. RESULTS: After 24 weeks of drug therapy, fluvastatin and pravastatin significantly reduced LDL susceptibility to oxidation as demonstrated by the reduced rate of oxidation (azo and Cu) and by prolonged azo-induced lag time (azo lag). The TC, LDL-C, and TG reduced significantly and HDL-C increased significantly. No differences between the drugs were observed. CONCLUSION: In hypercholesterolemic patients with CHD, both fluvastatin and pravastatin reduced LDL susceptibility to oxidation.

Lipid peroxidation and nitric oxide inactivation in postmenopausal women

OBJECTIVE: To assess the effect of endogenous estrogens on the bioavailability of nitric oxide (·NO) and in the formation of lipid peroxidation products in pre- and postmenopausal women. METHODS: NOx and S-nitrosothiols were determined by gaseous phase chemiluminescence, nitrotyrosine was determined by ELISA, COx (cholesterol oxides) by gas chromatography, and cholesteryl linoleate hydroperoxides (CE18:2-OOH), trilinolein (TG18:2-OOH), and phospholipids (PC-OOH) by HPLC in samples of plasma. RESULTS: The concentrations of NOx, nitrotyrosine, COx, CE18:2-OOH, and PC-OOH were higher in the postmenopausal period (33.8±22.3 mM; 230±130 nM; 55±19 ng/mL; 17±8.7 nM; 2775±460 nM, respectively) as compared with those in the premenopausal period (21.1±7.3 mM; 114±41 nM; 31±13 ng/mL; 6±1.4 nM; 1635±373 nM). In contrast, the concentration of S-nitrosothiols was lower in the postmenopausal period (91±55 nM) as compared with that in the premenopausal p in the premenopausal period (237±197 nM). CONCLUSION: In the postmenopausal period, an increase in nitrotyrosine and a reduction of S-nitrosothiol formation, as well as an increase of COx, CE18:2-OOH and PC-OOH formation occurs. Therefore, •NO inactivation and the increase in lipid peroxidation may contribute to endothelial dysfunction and to the greater risk for atherosclerosis in postmenopausal women.

Observações sôbre a anestesia geral pelo hidrato de cloral na prática da cirurgia veterinária

In this paper the A. A. report the observations about the general anesthesia by chloral hydrate on the veterinary surgery. The observations were made on emasculation practices of horses, mules and hogs. It was possible to establish the following conclusions: 1) The choral hydrate presents low cost, it harmless, and is of easy application. 2) The more recommendable dosis for equine and swine were : 12-13 g per 100 k of body weight, in destilled water solution at 30 and 20%, respectively. 3) The anaethestic was injected by intravenous way with good results; in horses and mules the applications were made in the jugular; in swine, in the anterior vena cava, as was described by Carle and Dewhirst, because it was impracticable in the ear vein. 4) The dosis applied produced deep narcosis not lasting to long and with no danger to the animal's life. 5) In the case of fattening hogs, it must be made a discount of about 40% on the body weight, to calculate dosis to be employed. 6) The tables A and B show the results, that may be considered as good.

Observações sobre parasitologia humana e veterinária em Mato Grosso

The most interesting observations were: 1 - The "mal de cadeiras" (believed to be an equine trypanosomose, due to Trypanosoma equinum) is a syndrome originated from malnutrition, overwork, intense blood-loss through blood-sucking arthropods (chiefly tabanids and bot-flies) and aggravated by the conditions prevailing in the "Pantanal" region of Mato-Grosso, when the waters of the Paraguay river basin ebb, after the periodic flood. The trypanosomes act only as one more factor contributing to the horses debility. 2 - The "curso preto dos bezerros" (calves black scour), usually attributed to poly-helminthoses, was observed to be related exclusively to pure Strongyloides papillosus infestations.

Emprêgo do teste químico para o estudo da eliminação das gonadotropinas durante o ciclo menstrual

A chemical test previously described for the diagnosis of pregnancy was applied to the study of the excretion of gonadotropin in the urine during menstrual cycle. The chemical test is based on the selective adsorption by kaolim of the reducing substances biologically related to urinary gonadotropin. The active substance when acidified to pH 4.0 is adsorbed by the kolin and eluated with O.1N sodium hydroxide. The alkaline solution is treated by Somogyi's copper reagent and the excess not reduced is titrated by 0.005 N sodium thiosulfate. Gonadotropin is quantitatively addorbed by kaolin at pH 4.0 and eluated by alkaline solution as previously demonstrated by the A. (1). In the present paper the complete menstrual cycle was studied daily. It was observed that normally there are two distinct maxima of excretion. This study is based on 11 normal cycles (24-30 days) and 34 abnormal ones. Normal cycles showed a intramenstrual estrogens elimination from 200 to 260 mice units determinated by the Allen - Doisy full estrus smear test. The abnormal cycles belonging also to normal women showed much less estrogen excretion (14 to 25 mice units) Table II). In those cases with decreased estrogen excretion no fall in the curve after 14 th. day was observed. The A. suggest that the peaks of gonadotropin excretion is not related to the oculation but possibly due, the first one, to the follicle stimulating hormone and the second to the luteinizing hormone of hormone stimulating of the inerstitial tissue.

Trypanosoma cruzi: antigen-receptor mediated endocytosis of antibody

Trypanomastigote forms of Trypanosoma cruzi were derived from tissue culture and incubated with immune and non-immune human sera. All immune sera showed high titers of specific humoral antibodies of the IgM or the IgG type. Agglutination and swelling of parasites were observed after incubation at 37ºC, but many trypomastigotes remained free-swimming in the sera for two to three days. The quantitiy of immune serum capable of lysing a maximum of 10 x 10 [raised to the power of 6] sensitized red cells was not capable of lysing 4 x 10 [raised to the power of 3] tripomastigotes. Typically, the parasites underwent cyclical changes with the formation of clumps of amastigotes and the appearance of epimastigote forms. Multiplication of the parasites was observed in immune sera. Further, the infectivity of the parasites to susceptible mice was not lost. All sera used produced similar general effects on the growth of the parasite. The antibody bound to T. cruzi appeard to enter cells by antigen-receptor mediated endocytosis. The ferritin-conjugated antibody was internalized and delivered to phagolysosomes where they might be completely degraded to amino-acids. This seemed to be a coupled process by which the immunoglobulin is first bound to specific parasite surface receptor and then rapidly endocytosed by the cell.

Study of an outbreak of cutaneous leishmaniasis in Venezuela: the role of domestic animals

During an outbreak of cutaneous leishmaniasis in a locality (Las Rosas, Cojedes State, venezuela) previously non-endemic, 12.9% of humans, 7% of dogs and 21.4% of donkeys (Equus asinus) had lesions with paraites. The agent in the three hosts was identified as Leishmania braziliensis, subspecies braziliensis at least in man and donkey. The probable vector was Lutzomyia panamensis. No infection was found in a small sample of wild mammals examined. The outbreak was apparently linked with the importation of donkeys with ulcers, from endemic areas. The Authors call attention to the fact that not only in the foci of "uta", but also in areas of the other forms of American cutaneous leishmaniasis, dogs are frequently found infected. They emphasize the necessity of searching for the infection in donkeys and of performing hemocultures and xenodiagnosis with sandflies in human, canine and equine cases, to verify their possible role as sources of infection, and not merely as dead ends in the epidemiological chain of the disease.

A simple method for assessing the binding of concanavalin A to mononuclear cell surfaces: no interference of visceral leishmaniasis serum on this binding

We report a simple method for evaluating the binding of concanavalin A (ConA) to human peripheral blood mononuclear cells (PBMC). The binding is evidenced by an immunoenzymic assay using peroxidase-conjugated immunoglobulins of a rabbit anti-ConA serum. Using the method we show that sera from patients with American leishmaniasis do not interfere with binding of ConA to PBMC.

Evolució i estabilitat d’àcids grassos i vitamines antioxidants a la llet materna

Recerca realitzada durant el periode Gener 2006-Abril 2007, a la Facultat de Farmàcia de la Universitat de Barcelona sota la direcció de la Dra. M. Carmen López Sabater. La nutrició durant els primers anys de vida té una enorme importància, amb repercussions en el creixement i desenvolupament del nen i en les possibles malalties futures. Tenint en compte que la llet materna és l’aliment ideal per al nadó durant els primers sis mesos de vida, és important conéixer la composició nutricional de la mateixa i la seva evolució al llarg de la lactància (calostre, llet de transició i madura). La llet conté nutrients majoritaris (proteïnes, carbohidrats i lípids) i nutrients minoritaris, com vitamines i minerals. S'ha desenvolupat i validat un mètode per a la determinació d'àcids grassos (AG) i Àcid Linoleic Conjugat (CLA) en llet materna per Cromatografia de Gasos "fast" (fast-GC), permetent reduir el temps d'anàlisi considerablement en comparació amb els mètodes utilitzats fins el moment. La repetibilitat i reproducibilitat trobades han estat bones, amb coeficients de variació inferiors al 10% en tots els casos. L'aplicació a 6 mostres de llet materna ha demostrat que es tracta d'un mètode senzill, ràpid, eficient i pràctic per a l'anàlisi rutinari d'un elevat nombre de mostres. També s'ha començat a desenvolupar un mètode per a l'anàlisi dels anàlegs de tocoferol de la llet materna, degut a l'interès de la vitamina E com a antioxidant natural. És un mètode per Cromatografia Líquida d'Ultra Resolució (UPLC) amb detecció per Fluorescència (FD) i per Photodiode Array (PDA) que permet treballar amb quantitats de mostra petites mantenint o inclús millorant la sensibilitat i reduint el temps d’anàlisi.

Zoonotic cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis associated with domestic animals in Venezuela and Brazil

After outbreaks of cutaneous leishmaniasis in Solano State, Venezuela, 5% of the population had parasitized ulcers while after similar outbreaks in Mesquita, Rio de Janeiro State, Brazil, 9% had the disease. In these foci children, including some under six years of age, wre affected. There was no significant difference in the occurence of the disease according to sex or type of employment. In Solano, 3% of dogs and 28% of donkeys had parasitized lesions, while in Mesquita these indices were 19.8% and 30.8% respectively. The parasite from man, dogs and equines was identified as Leishmania (Viannia) braziliensis, by zymodeme and serodeme characterization. In these foci there is evidence suggesting that leishmaniasis is a zoonosis, possibly with equine and dogs as reservoirs, although both a wild enzootic cycle and the role of man as a source of infection can not be ruled out. Transmission is assumed to occur peridomestically by sandfly vectors such as Lutzomyia panamensis in Venezuela and Lutzomyia intermedia in Brazil. Information about the origin of these foci suggests that infected equines may be an important factor in the dissemination of the parasite in a peridomestic situation where these sandflies are abundant.