Commiphora leptophloeos (Mart.) J.B. Gillet, from the Burseraceae family, is a plant
native to Brazil, found in the Caatinga, Cerrado, and Amazon biomes. It is popularly
known as imburana and is recognized for its medicinal properties in treating general
inflammation. Moreover, there are reports of its effects in alleviating gastrointestinal
symptoms such as colic and diarrhea. This plant contains important secondary
metabolites, including flavonoids such as orientin, vitexin, and quercetin, which have
recognized intestinal anti-inflammatory activity. Thus, this study aimed to evaluate the
effects of the free and nanoparticulated hydroethanolic extract (NPE) from C.
leptophloeos leaves in models of intestinal inflammation (preventive and curative) and
colitis-associated colorectal cancer (CAC). Additionally, this study included the
phytochemical evaluation of the extract and the development and characterization of a
nanoformulation containing the extract. Phytochemical annotation identified Cglycosylated flavonoids (quercetin, isovitexin, isoorientin, and orientin) and Oglycosylated flavonoids (quercetin-O-(O-galloyl)-hexose and quercetin-O-feruloyl-Ohexose), along with quinic acid. The total phenolic and flavonoid contents were 311.02 ±
0.03 mg/g and 174.90 ± 0.02 mg/g, respectively. The physicochemical characterization
of the NPE demonstrated a size of up to 100 nm, a PdI value of 0.13, stable Zeta potential,
and a significant reduction in the effective dose for regulating inflammation and
tumorigenesis signaling pathways—from 400 mg/kg with the free extract to only 4 mg/kg
with the NPE. Treatment with the free extract showed significant results (p<0.05) in
regulating inflammatory markers such as NF-κB p65/Cox-2, TNF-α, IL-1β, IL-6, Il-17,
and mTOR in a colitis model induced by 2,4-dinitrobenzenesulfonic acid (DNBS). The
free and nanoparticulated extract in a colitis model induced by dextran sulfate sodium
(DSS) regulated Tlr2/4, Jnk1/2, Cox-2, Tnf-α, Il-1β, Il-6, Il-17, Mip2, and Icam-1. In both
models, improvements in clinical symptoms and intestinal cytoarchitecture were
observed. Furthermore, both the free extract and NPE exhibited antitumor activity in the
CAC model, acting on important tumorigenesis signaling pathways such as Tp53, Ctnnb1,
Bax, and Bcl-2, reducing tumor volume, and increasing the abundance of the bacterium
Akkermansia muciniphila. In conclusion, C. leptophloeos exhibits both preventive and
curative intestinal anti-inflammatory activity in experimental models, as well as
antitumor activity. These findings support the potential development of a new
phytoingredient for the treatment of chronic inflammatory diseases.

Kalanchoe pinnata (Lam.) Pers. is a plant native to Madagascar that can be found in

Brazilian biomes such as the Caatinga. It has a complex chemical composition with

considerable potential as a raw material for use in cosmetic products. The purpose of

this study was to develop safe, multifunctional cosmetic formulations with antioxidant,

antityrosinase, and moisturizing activities for the prevention of signs of skin aging,

enriched with the aqueous extract of Kalanchoe pinnata leaves. The extract was

chemically characterized through the quantification of sugars, phenolic compounds,

and flavonoids, and its safety was evaluated using 2D and 3D in vitro models, in vivo

and clinical assessments. The antioxidant potential was assessed in vitro and in vivo

using Caenorhabditis elegans as a model organism. Antityrosinase activity was

determined using an enzymatic method. Nanoemulsions were developed based on the

study of the hydrophilic-lipophilic balance using the phase inversion method and were

subjected to preliminary and accelerated stability tests. A high-performance liquid

chromatography method was developed and validated for the quantification of the

marker compound in the nanoemulsions, which were evaluated for safety and clinically

regarding moisturizing efficacy and skin microrelief. The extract contains flavonoids,

phenolic compounds, and sugars, and demonstrated safety for topical use, supporting

the claim "dermatologically tested." It acted at different stages of the oxidative cascade

in vitro and in vivo at the evaluated concentrations and showed antityrosinase activity.

The developed formulations remained stable for 30 days under different storage

conditions, and the quantification of the Bp1 marker demonstrated the chemical

stability of the nanoemulsions, which proved to be safe and effective in moisturizing

and anti-aging benefits. Thus, this work provides scientific contributions and

technological innovation to the cosmetic sector, while also adding value to the plant's

production chain, with the goal of enhancing social development, with the aim of

valuing social aspects, boost the bioeconomy and Brazilian biodiversity.

Kalanchoe pinnata (Lam.) Pers. is a plant native to Madagascar that can be found in

Brazilian biomes such as the Caatinga. It has a complex chemical composition with

considerable potential as a raw material for use in cosmetic products. The purpose of

this study was to develop safe, multifunctional cosmetic formulations with antioxidant,

antityrosinase, and moisturizing activities for the prevention of signs of skin aging,

enriched with the aqueous extract of Kalanchoe pinnata leaves. The extract was

chemically characterized through the quantification of sugars, phenolic compounds,

and flavonoids, and its safety was evaluated using 2D and 3D in vitro models, in vivo

and clinical assessments. The antioxidant potential was assessed in vitro and in vivo

using Caenorhabditis elegans as a model organism. Antityrosinase activity was

determined using an enzymatic method. Nanoemulsions were developed based on the

study of the hydrophilic-lipophilic balance using the phase inversion method and were

subjected to preliminary and accelerated stability tests. A high-performance liquid

chromatography method was developed and validated for the quantification of the

marker compound in the nanoemulsions, which were evaluated for safety and clinically

regarding moisturizing efficacy and skin microrelief. The extract contains flavonoids,

phenolic compounds, and sugars, and demonstrated safety for topical use, supporting

the claim "dermatologically tested." It acted at different stages of the oxidative cascade

in vitro and in vivo at the evaluated concentrations and showed antityrosinase activity.

The developed formulations remained stable for 30 days under different storage

conditions, and the quantification of the Bp1 marker demonstrated the chemical

stability of the nanoemulsions, which proved to be safe and effective in moisturizing

and anti-aging benefits. Thus, this work provides scientific contributions and

technological innovation to the cosmetic sector, while also adding value to the plant's

production chain, with the goal of enhancing social development, with the aim of

valuing social aspects, boost the bioeconomy and Brazilian biodiversity.

Keywords: Biodiversty. Bryophyllum pinnatum. Cosmetics. Moisturizer. Saião.

Chagas disease, also known as American Trypanosomiasis, is a neglected tropical illness
caused by the hemoflagellate protozoan Trypanosoma cruzi. Currently, only two drugs,
Benznidazole (BNZ) and Nifurtimox, has been approved for disease treatment, with BNZ
being the only one authorised for use in Brazil. However, BNZ faces various limitations
including low bioavailability and higher toxicity to the host. Parasite resistance to the
treatment has been also reported. In this regard, nanotechnological formulations for drug
delivery have emerged as a promising alternative to improve the pharmacological
properties and bioavailability of this drug. In this study, we have investigated the in vitro
effectiveness of cationic PLGA-cholesterol hybrid loaded with BNZ as a drug-delivery
platform against T. cruzi. The nanoprecipitation method was used to obtain the HNPs,
and their size, polydispersity index, and zeta potential were determined. Following PEI
functionalization, the size of the NPHs was less than 200 nm, with homogeneous particle
distribution and a zeta potential ranging from -20 to +10. The efficiency of encapsulation
was determined to be 40% for anionic HNPs, increasing to 54% for cationic counterparts.
FTIR and XRD analyses corroborated the successful encapsulation of BNZ.
Morphological characterization throughout scanning electron microscopy (SEM)
demonstrated spherical-shaped particles, regardless of their composition. In vitro studies
showed that nanoparticles are safe at the evaluated concentrations for cardiomyoblasts.
Furthermore, compared to the free drugs, HNPs exhibited higher trypanocidal effects
against intracellular amastigotes, as evidenced by IC50 values of <5.9 and 38.6,
respectively. However, no statistical difference was observed between the trypanocidal
activity of the anionic and cationic systems. Fluorescence microscopy images showed the
internalization of HNPs from 5 minutes onward. Taken together, our results showed the
successful development of HNPs, underscoring their potential as a promising platform
for the delivery of therapeutic agents used in the treatment of Trypanosoma cruzi
infection.

Solid-state modification technologies such as salts and cocrystals represent
multicomponent systems that allow improvement of physical-chemical properties of
drugs, such as stability and aqueous solubility. Ferulic acid (FEA) is a molecule with
high antioxidant and hypoglycemic potential, but with limited aqueous solubility, which
may imply low bioavailability. Metformin (MT) is a consolidated hypoglycemic agent in
the treatment of type 2 diabetes mellitus (DM), which showed a synergistic effect with
AFE. The objective of this study is to develop and characterize multicomponent
systems with AFE and MT with improved aqueous solubility, as well as to incorporate
them into a solid pharmaceutical form. For this, a recrystallization process
demonstrated high reproducibility and yield (91.3 ± 0.8%). After a series of
experiments, it was possible to develop a multicomponent system between FEA and
MT in a 1:1 molar ratio, characterized as a salt, SFM (metformin ferulate, by X-ray
diffraction techniques, thermal techniques, infrared spectroscopy and microscopy.
FEA in SFM showed an increase of at least 740 fold in its aqueous solubility (643 ± 18
mg/mL). Tablets were developed with the SFM, which met the quality control
requirements for this dosage form. From in vitro dissolution tests with SFM tablets with,
a superior dissolution efficiency was calculated, in relation to a control formulation, of
95.4 ± 0.5% and 42.1 ± 0.5%, respectively. The FEA and MT contents were determined
using a high-performance liquid chromatography method, which was properly
developed and validated. Finally, in a stability study at 40 ºC for 3 months, no physical
or chemical changes were found, for both the SFM raw material and tablets. Thus, the
developed product emerges as a technological innovation to be applied in the
treatment of DM.

Chagas disease is caused by the parasite Trypanosoma cruzi and is considered

a neglected tropical disease. Novel immunoadjuvants have been studied to

improve the immune response and treatment efficacy. Nanoparticles are drug

delivery devices that target specific cells, enhancing therapeutic action and may

possess immunomodulatory characteristics. This study explored the use of

cationic PLGA-PEI nanoparticles as potential immunoadjuvants in response to

T. cruzi infection. The results obtained were satisfactory and promising. During

the research, it was demonstrated that cationic nanoparticles interacted with

proteins and exhibited favorable properties, such as appropriate size and

cellular viability. When tested as immunoadjuvants in animal models, the

nanoparticles stimulated an immune response similar to aluminum hydroxide, a

commonly used human immunoadjuvant. Furthermore, the nanoparticles were

observed to stimulate the production of high avidity and specific antibodies of

subclasses IgG2b and IgG3, which play important roles in the inflammatory

process and effector cell activity. These findings are relevant as the formulated

nanoparticles are unprecedented in the literature and demonstrate significant

potential for the development of immunomodulatory strategies in the context of

T. cruzi infection.

Moringa oleifera Lam. (Moringaceae), popularly known as moringa or white acacia, native to
India, is currently cultivated in several countries around the world, and distributed in many
regions from Brazil. It is a fast growing perennial plant, with significant content of nutrients
such as vitamins, minerals, amino acids, and proteins. All parts of the plant are utilized (seeds,
leaves, fruits, branches and roots) medicinally, ornamentally, industrially or for food for human
and animal use. It can also be an alternative source for the improvement of drinking water
quality and for fertilization in the cultivation of other plant species. However, when it comes to
therapeutic use, the leaves are the most used part of the plant, mainly to prevent and treat
diabetes. However, the problem regarding the therapeutic use of M. oleifera is that there is no
definition about the chemical composition content to ensure the safe and effective use of
derivatives of the species. Within this context, the present work aimed to standardize the culture
conditions and optimize the extraction conditions to obtain a flavonoid-rich extract from M.
oleifera leaves, evaluate the in vitro and in vivo toxicity and the in vitro anti-inflammatory
activity. It was also part of the thesis to compare different samples of moringa commercialized
in Brazil and the United States. For this, initially the study of different soil cultivation
conditions (mineral fertilization, mineral fertilization + organic compost, mineral fertilization
+ charcoal, and the control, soil without fertilization) and two harvesting seasons (dry and rainy)
of the leaves of the species M. oleifera was developed, aiming to identify the best condition to
obtain the highest content of flavonoids in the leaves. The extraction process was then
optimized to concentrate the flavonoids. This optimized extraction method was applied to
samples from different growing and harvesting conditions to select the most promising sample.
All extracts were submitted to analysis of total content of phenols, flavonoids, sugars and
proteins, yield analysis and chromatographic profile by ultra efficient liquid chromatography
coupled to mass spectrometer (CLUE-EM/MS). The extract from the condition of cultivation
with mineral fertilization was the one with the highest content of phenols, flavonoids, proteins
and good yield, being selected for the next steps of the study. In the analysis by CLUE-EM/MS,
it was possible to characterize eith majority substances predominantly from the flavonoid class,
with prevalence of flavonol type nuclei. Cytotoxicity and oral toxicity (acute and repeated
doses) of the optimized flavonoid-rich extract were evaluated. And was observed safety for
acute use until a dose of 2,000mg/kg and subchronic use at a dose of 400 mg/kg. Finally, a
comparative analysis was made of the extracts of all culture conditions against commercial
samples that were purchased in Brazil and the United States, and a comparison between plant
drug (pulverized dried leaves) and plant derivative (plant extract). A visual comparison was
made, of chemical profile by CLAE-DAD and by in vitro biological activity, antioxidant and
anti-inflammatory, with two macrophage cell lines. The results showed great variation visually
and between the phytochemical profiles of the commercial samples, regardless of the country
of their commercialization. And the phytochemical and in vitro activity evaluation showed
better results for the vegetal extract samples, compared to the vegetal drugs. This set of results
contributes to the quality control of the optimized extract of M. oleifera leaves, to the future
characterization of the species' chemical markers and to the development of a with standardized
process to obtain products with quality, efficacy and safety for the population.

;

The genus Selaginella, the largest genus of Lycopodiophyta comprise about 700-
750 species, which several species are recognized for their pharmacological activities.
Our overview of the phytochemical and bioactive aspects pointed out that future studies
should afford valuable new data on better explore the biological potential of the flavonoid
amentoflavone and their derivatives as chemical bioactive entities; develop studies about
toxicity and, finally, concentrate efforts on elucidate mechanisms of action for biological
properties already reported. The family Selaginellaceae managed to survive the many
biotic and abiotic pressures during the last 400 million years. Several species of the genus
Selaginella have been acquired an interesting evolutionary trait: desiccation tolerance
(DT). Selaginella convoluta is a desiccation-tolerant plant native from Brazilian semiarid.
DT plants have highly effective drought-resistance mechanisms that allow their survival
under extreme conditions. In this study we investigate Selaginella convoluta in two
hydration states: (i) dehydrated – harvested in the natural semiarid environment; and (ii)
hydrated – after acclimatization in lab. The proteome dataset pointed out that shoots and
roots appear to have different strategies in response to different hydration states; however,
appear to have a well-orchestrated response to regulate desiccation tolerance. The
presence of enzymes involved in the biosynthesis of specialized metabolites such as
phenolic, terpenoid, as well as metabolites containing nitrogen/sulfur were identified
through proteomic analysis. The NMR-based metabolite profiling showed the presence
of metabolites such as carbohydrates and phenolic derivatives related to desiccation
tolerance events, corroborating with the proteome data. As an unprecedented finding, the
most important annotated alkaloid was anabasine. The specialized metabolites,
flavonoids, bioflavonoids and selaginellins were the main phenolic derivatives annotated.
Thus, we performed a LC-MS/MS analysis combined with the molecular networking
(MN) approach to assess the chemodiversity of flavonoids and selaginellins. So far, this
has been the first study on Selaginella spp. using the MN approach that showed not only
a proposal of the fragmentation pattern for all the selaginellins annotated, but also the
prediction of new selaginellins based on the MS/MS spectral data. Several biological
potentials of selaginellin from Selaginella spp. have been reported. Selaginellins are
naturally produced at low levels in the plant, which makes the chemical extraction of
these metabolites difficult and time consuming. In this context, methyl jasmonate (MeJA)
has been used to induce specialized metabolites of pharmaceutical interest. Our data
showed that MeJA can be an interesting alternative to induce the accumulation of
selaginellins in S. convoluta. In addition, the in silico studies pointed out interesting
bioactive potentials for selaginellins: antineurodegenerative, antiproliferative and
antiparasitic. The MN approach allowed the prediction of seven unprecedent
selaginellins, which contribute as a new chemical information for this species.

Mansoa hirsuta D.C. (Bignoniaceae), popularly known as cipó-de-alho, is a native species from
the Brazilian semiarid region and is traditionally used in the treatment of sore throats and
diabetes. Studies report that extracts and fractions of this species have several pharmacological
actions such as antifungal and antioxidant activities, endothelium-dependent vasodilator effect,
and inhibition of the angiotensin-converting enzyme, nitric oxide production, and lymphocyte
proliferation. In this scenario, this study followed research lines: (1) investigating the
composition, toxicity, anti-inflammatory and analgesic actions, and chemical of the fraction
obtained from M. Hirsuta leaves (MHF); and (2) developing chitosan films incorporated with
MHF as an innovative formulation for treating wounds. In vitro cell viability was evaluated in
3T3 cells using the MTT assay. At 24, 48, and 72 h, MHF did not show damage to cell viability.
For the acute toxicity assay, a single dose of the fraction was administered orally in male and
female mice, with no mortality or signs of toxicity. Additionally, in the subchronic toxicity test,
no relevant toxicological changes were observed. The results of the open field and rota-rod tests
showed that MHF did not alter the locomotor activity and behavior of the mice, nor did it alter
the motor coordination and balance of the treated animals. In evaluating the anti-inflammatory
activity, single oral administration of MHF significantly reduced carrageenan-induced edema
and levels of myeloperoxidase. Furthermore, it has decreased the migration of leukocytes and
proteins in the air pouch model. Regarding the antinociceptive activity, it was observed that the
highest doses reduced abdominal contortion in response to the administration of acetic acid and
significantly inhibited the second phase of the formalin test. Ultra-performance liquid
chromatography coupled with a time-of-flight analyzer and mass spectrometry indicated that
this fraction is rich in acid triterpenes that can be derived from oleanolic and ursolic acids. Then,
films were prepared using the solvent evaporation method and characterized by Fouriertransform infrared spectroscopy, X-ray diffraction, thermogravimetry, differential scanning
calorimetry, scanning electron microscopy, and atomic force microscopy. In addition, tensile
strength, elongation at break, and thickness were also measured. Chitosan films containing
MHF (CMHF) exhibited characteristic bands of chitosan and MHF, revealing a physical
mixture of both. CMHF showed amorphous nature, thermostability, and MHF dispersion in the
chitosan matrix, resulting in a rough structure. Incorporation of the fraction into the chitosan
matrix significantly improved the mechanical performance and thickness of the films.
Treatment of wounds in vivo with CMHF for seven days showed a characteristic area of
advanced healing, re-epithelialization, cell proliferation, and collagen formation. Furthermore,
wound closure reached 100% contraction after ten days of treatment with modulation of
interleukins. Therefore, the results of this study indicated that MHF is rich in acid triterpenes
and an anti-inflammatory and analgesic potential without causing acute or subchronic toxic
effects, and could be a promising source to be explored. Furthermore, the incorporation of the
M. hirsuta fraction in chitosan films was advantageous and showed great potential to stimulate
healing and regeneration of wounds.

The resistance to conventional drugs in diseases of infectious and neoplasic origin is
a serious public health problem worldwide, presenting high morbidity and mortality,
leading to an incessant search for new therapeutic agents in different natural
sourcesIn this context, the scorpion venom constitutes a rich arsenal of bioactive
molecules, being non disulphide-bridged peptides (NDBPs) target of studies due to
its multifunctional actions.Stigmurin (FFSLIPSLVGGLISAFK-NH2, 1795.2 Da) and
TsAP-2 (FLGMIPGLIGGLISAFK-NH2, 1733.2 Da, NDBPs obtained from the venom
gland transcriptome of the Tityus stigmurus scorpion with charge +2 and theoretical
isoelectric point of 8,75, have demonstrated antimicrobial and antiproliferative effect
in vitro and antibiotic action in the polymicrobial sepsis model. Considering the
multifunctional action of NDBPS, in this study the three-dimensional structure of the
Stigmurin was elucidated by nuclear magnetic resonance and the antiproliferative,
immunomodulatory, antiparasitic, antiviral, antioxidant and antibiofilm activity of the
peptides Stigmurine and TsAP-2 were evaluated. The healing and antibiotic effect of
both peptides has been evaluated by bacterial wound infection model in vivo, as well
as expanding the in vitro antibacterial potential of the TsAP-2. Stigmurin in
trifluoroethanol: water showed a random conformation in the N-terminal region, with a
helical structure from the seventh amino acid residue, being the first NDBP present in
the venom of scorpions of the genus Tityus with an elucidated three-dimensional
structure. Stigmurin and TsAP-2 revealed antiproliferative action on neoplastic cells
in non-toxic concentration for normal cells, modulating the release of nitrite in murine
macrophages stimulated by lipopolysaccharides. Stigmurin and TsAP-2 did not
demonstrate a leishmanicidal effect on the amastigote form of Leishmania
braziliensis. However, TsAP-2 revealed a high trypanocidal action of on epimastigote
and trypomastigote forms of Trypanosoma cruzi, with broad spectrum of antibacterial
action against Gram-positive and Gram-negative microorganisms. A protective effect
on primate renal epithelial cells against Zika virus infection has also been
demonstrated in the pretreatment assay for both peptides, revealing a high virucidal
action on herpes simplex virus type 1, with antiviral activity in post-infection assay. In
addition, Stigmurin and TsAP-2 were able to scavenge the hydroxyl radical in vitro at
concentration of 10 µM and 5 µM, respectivety, reducing its action in high
concentrations, indicating the formation of agglomerates. Both peptides showedantibiofilm activity, as well as antibacterial action in the skin wound infection model,
increasing the retraction rate of the lesion, suggesting the ability to induce tissue
repair. In histopathological analysis, TsAP-2 promoted increased neovascularization
and re-epithelization, reducing necrosis in the surgical wound. Taken together, the
data suggest that Stigmurin and TsAP-2 are promising multifunctional peptides for
use as prototype to obtaining new therapeutic agents.

The non-clinical study is a primordial step of the research and development process
of new drugs, ensuring the safe use and the efficacy in humans. The isatin
derivatives have aroused interest for their diversity of biological activities (antiviral,
antibacterial, antitumor, anti-inflammatory, analgesic, etc.), with the possibility of
being models of new therapeutic agents. So, the aim of this study was to evaluate, in
vivo, the toxicity and the anti-inflammatory and antinociceptive activities of the isatin
thiosemicarbazone (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl
hydrazinecarbothioamide or PA-Int6. Swiss mice of both sexes were used to
evaluate three doses of this compound, administered by gavage: 1.0 mg/Kg (Test 1);
2.5 mg/Kg (Test 2) and 5.0 mg/Kg (Test 3). A prior assessment of central neural
activity was made through the Open Field and Rotarod tests. Toxicity assays (acute
and subchronic) performed were based on OECD protocols (Nos. 423, 407 and 408).
In order to evaluate the anti-inflammatory activity were used the carrageenan
induced paw edema model and the zymosan-induced air pouch model. For the
antinociceptive activity, the formalin test and the acetic acid-induced abdominal
writhing test were used. Impairment on locomotor activity and coordination of animals
treated with PA-Int6 (5.0 mg/Kg) has not been demonstrated. In addition, during the
toxicity assays, there were no deaths or physical and behavioral changes. In the
acute toxicity, no hematological parameter presented any difference, whereas the
increase of Triglyceride for males was the only biochemical alteration. In subchronic
toxicity few biochemical parameters had increased values among males (Albumin
and Total Proteins) and among females (Triglyceride and Albumin). The paw edema
test demonstrated that since the first time evaluated all test groups showed
inflammatory edema below 50%, but in the second and fourth hour they had
significant responses. On induced inflammation by the airbag model there was
decrease both in the leukocyte migration (55%, 50% and 47%) as in the protein
extravasation (71%, 58% and 71%) for the three doses, respectively.
Antinomyciceptive activity was observed for the three doses in the second phase of
the formalin test, with reduction of paw pain time of 73.61% (Test 1), 79.46% (Test 2)
and 73.85% (Test 3 ). The number of abdominal writhes decreased with increasing
dose of PA-Int-6, but only with 5.0 mg/kg there was a significant response, reducing
24.88%. Thus, the results of this study suggest that PA-Int6 is safe, because it has
demonstrated few and slight alterations, which can be physiologically reversed. With
anti-inflammatory and antinociceptive activities, this compound can be considered
promising, with the prospect of following in the research and development process as
a candidate for a new drug.

Olanzapine is a benzodiazepine which proved efficacy in the treatment of positive and
negative symptoms of schizophrenia, bipolar disorder and others psychoses. The
pharmaceutical industry and analytical laboratories have focus on quality, safety and efficacy
of their products, for this planning tools and monitoring are required in the production scale.
The concept of Process Analytical Technology introduces innovative approaches, as
improvement of process control and comprehension of carried out procedures to improve the
efficiency.
This study applies the near infrared spectroscopy to build the Process Analytical
technology in the production of Olanzapine coated tablets, since this technique has being
wide use, because it is not necessary to prepare the samples, the absence of residues
produced and the analysis are fast. Previously to implementation of Process Analytical
Technology some laboratory steps are required to evaluate if the technique are sensitive to
the objective. To prove the theory, alternative methods were developed to compare with their
references. In the Chapter 1 it was carried out the assay of the drug using high performance
liquid chromatography and near infrared associated to multivariate calibration techniques
(partial least square – PLS), after the spectra pre-treatment). The results obtained by the
near infrared were compared to the chromatography and as response: the assay by both
methods did not presented statistically significance difference. In the Chapter 2, the thesis
was focused at the construction of multivariate control charts, based on Net Analytical Signal
(NAS) to monitor the assay of the drug in laboratory and production samples using near
infrared spectroscopy. The control charts (NAS, interferers and residuals) were built and
they were promisor with high level of recognition (95 to 100%) of samples that were exactly
in or out of the specified range. Lastly, at the Chapter 3 the determination of water content by
near infrared associated to multivariate calibration, PLS, of Olanzapine and excipients
granulate mixtures in laboratory and production samples was analyzed and compared to the
reference method (Karl Fischer). The results, when NIR and Karl Fischer were compared,
presented no significant statistically difference, promoting the NIR associated to multivariate
calibration also works to analyze the water content into the coated olanzapine tablets.
The results showed that the multivariate calibration models constructed, using the
spectra obtained by the near infrared, were promisors, when compared to the reference
methods. Therefore, the alternative analytical tool (near infrared spectroscopy) can be
installed at production scale to monitor the drug quality at this matrix used.

Chemical composition and therapeutic properties of Aloe vera (L.) Burman f. (A.
vera) explain its potential use in cosmetic, nutritional and pharmaceutical applications.
Mucilaginous gel present in the leaves of A. vera is rich in several compounds, mainly
polysaccharides. Most of the reported therapeutic activities of these polysaccharides are
indicated by the presence of glucomannan, with acemannan being the predominant
polysaccharide. Acemannan has been incorporated into commercially available wound
healing products for wound application. The objective of the present work has
developed a hydrogel for topical use containing mucilage of A. vera to use in skin
disease like psoriasis. The hydrogels were prepared with 80% (w / w) of A. vera
mucilage, varying two types of polymer (anion and nonionic) carbomer 940 1% (FC1
and FC2) or hydroxyethylcellulose 2% (FH3 and FH4), as well as preservatives,
antioxidants and sequestrants. Polysaccharide fractions were extracted from the
mucilage. All of them were used as a group of chemical markers and characterized by
X-ray diffraction, infrared (IR) spectroscopy and nuclear magnetic resonance (NMR).
The quantification of these markers in the raw material, mucilage, and in the hydrogel
was carried out using spectrophotometric techniques in the UV-VIS region. Four
hydrogels (FC1, FC2, FH3 and FH4) were obtained from the mucilage of A. vera and
evaluated for their organoleptic, rheological, pH and acemannan content. The hydrogels
FH3 and FH4 (hydroxyethylcellulose) presented acemannan content of 6.76 and 4.01
mg / g, respectively, FH4 presented pH 4.6 and FH3 showed reoptic behaviour. The
carbomer formulations FC1 and FC2 presented acemannan content of 8.69 mg / g and
9.17 mg / g, respectively, ideal pH for application in psoriasis, good spreadability,
rheological behaviour of the pseudoplastic and thixotropic gels, therefore, taking into
account the characteristics ideal for a semi-solid form of topical application.

Chagas disease is a kind of Neglected diseases, it represents a global health problem due to
the industry's lack of interest in research for new drugs. Benznidazole (BNZ), a drug available
in Brazil for the treatment of Chagas disease, and distributed by governement, presents some
limitations regarding its use, especially in the chronic's phase. Pharmaceutical technology
provides for the development of systems that increase drug solubility or increase its
concentration in infected cells / tissues which directly reflects the increased bioavailability
and therapeutic efficacy of BNZ. Thus, colloidal lipid systems (CLS), such as emulsions,
microemulsions and nanoemulsions, represent an interesting alternative for the increase of
oral and parenteral bioavailability of BNZ. This work aims to improve the method of
obtaining these systems and characterize the biocompatible CLS containing medium chain
triglycerides (Miglyol®812) stabilized by a suitable mixture of surfactants (soy
phosphatidylcholine and sodium oleate) associated with a co-solvent, besides evaluating the
trypanocidal effect of these systems. The improvement of the ideal conditions for obtaining
the systems of interest was evaluated by the study of the phase inversion temperature and the
addition of a new component, a co-solvent. The validation of the methodology followed the
guide recommended by ANVISA and "ICH". Liquid and translucent systems were formed
with addition of 2-methylpyrrolidone (NMP) as co-solvent with uniform droplet size and less
than 162.66 nm, when more than 5% of the surfactant mixture was added. The systems
remained stable for up to 60 days when stored at room temperature. When stored at 45 ° C,
the stability time was shorter, with a direct relationship with the co-solvent concentration, the
presence of BNZ and sialic acid Added. The rheology measurements showed a behavior
dependent on the amount of co-solvent added to the system. Polarized light microscopy and
low-angle X-ray scattering images confirmed the formation of isotropic systems when the
cosolvent is in a concentration greater than 5%. Atomic force microscopy images showed the
formation of large numbers of spherical, smooth droplets of a size well correlated with the
measurements obtained by the dynamic scattering of light. Addition of sialic acid to the
nanoemulsion containing 10% NMP did not increase drug uptake. The release of BNZ in
systems containing sialic acid at different concentrations was up to 3 (three) times faster than
systems without the substance. The mathematical model that fit all the systems investigated
was the Kormeyer-Peppas (r2 > 0.911), indicating mechanism non-Fickian of release. The
systems present biocompatibility in different cell lines (Vero, SiHa and LLC-MK2) over a
long period of exposure, up to 72 hours. The anti-epimastigote activity showed that much

smaller amounts of BNZ are able to inactivate this parasite life form when it is associated
with NMP-containing systems. The trypomastigote forms of the parasite showed greater
sensitivity to the constituents of the investigated systems and, cell death in concentrations up
to 170 times lower than when the drug was used alone and not incorporated in nanoemulsions.
Nanoemulsions have the ability to transport BNZ in biocompatible systems with excellent
trypanocidal activity, thus representing a promising future in the treatment of Chagas disease,
considering that to date there are no liquid dosage forms of administration which make it
impossible to adjust the dose, decrease the side effects.

ABSTRACT: Current costs of treating patients with systemically progressing
inflammatory diseases, especially sepsis, are the leading cause of death in non
cardiological intensive care units worldwide. In Brazil, the incidence of sepsis in
patients is becoming more frequent. Despite pharmacological, technological
and surgical advances, mortality due to sepsis and/or associated diseases
remains high worldwide, hence the search for easily accessible and inexpensive
therapeutic alternatives to contain the progression of this disease. Natural
products have played an important role in the pharmaceutical industry because
some of these substances act in a beneficial way on the human immune
system. In recent times, there is a growing investigation of the possible
biological and therapeutic properties attributed to bullfrog oil, since this oil has
been used indiscriminately by the population in the treatment of various
diseases, such as bronchitis, asthma, lichen sclerosus, furunculosis, sebaceous
cyst, and for healing of skin and mucous membranes. However, the possible
consequences of excessive consumption of this oil are increased production of
eicosanoids derived from proinflammatory arachidonic acid and deficiency in
hepatic regulation, predisposing to steatosis, which may progress to hepatic
inflammation and fibrosis. Many hypotheses are based on the performance of
the compounds present in bullfrog oil in the modulation of the inflammatory
response, thus preventing the onset of tissue injuries. The microemulsion is
presented as a possible alternative to a new drug delivery system, in order to
reduce the incidence of hepatotoxicity and protect the organism against the
installation of tissue lesions as a function of the septic condition. The present
study is aimed to evaluate the anti-inflammatory potential of pure bullfrog oil and
a microemulsion system in an experimental model. In this study, a
microemulsified system (WIV) was determined and characterized, and applied
in biological tests to evaluate the anti-inflammatory potential of pure oil and
microemulsion. We used the sepsis model, induced by cecal ligant puncture
(CLP) technique, and the formalin-induced muscle injury. The tests were
performed in murine models, where the animals were randomly separated into
groups and treated with pure bullfrog oil and microemulsion, using the gavage
technique for further evaluation of the hepatotoxic and anti-inflammatory
potential. For the analysis of the anti-inflammatory potential in a sepsis model,
bronchoalveolar lavage was performed with subsequent inflammatory cell
counts and histopathological analyzes of lung tissue. For the analysis in the
muscle injury model, the horizontal extension of the limbs of the animals was
evaluated, as well as the histopathological analysis of the muscle tissue. For the
analysis of the hepatotoxic potential of the substances, the post-sepsis survival
rate and the histopathological analyzes of the hepatic tissues of the animals
were evaluated. When evaluating the toxicity of pure bullfrog oil and in a
microemulsion system, it was observed that in the group that received the microemulsion (ME), the liver had the architecture preserved, but with clinical
signs of hepatic steatosis, unlike the pure bullfrog oil group, which presented
multiple outbreaks of hepatocytic necrosis accompanied by polymorphonuclear
infiltrates. These findings indicate a picture of steatohepatitis, that is, a more
advanced stage and a precursor of hepatic carcinoma. When evaluating the
survival of the animals, it was observed that the ME group survival rate was
significantly higher when compared to the oil group. When evaluating the anti
inflammatory potential of the ME and the oil group in a sepsis model, the
potential for modulation of the inflammatory response was observed in both
groups, since the leukocyte migration to the lungs was significantly reduced
(P<0.01) after the induction of sepsis. When analyzing the histology of the lung
tissues of the animals of both groups, intense wear was observed in the animals
of the oil group, when compared with the ME group, where there was little
tissue compromise. In the muscle injury test, it was observed that the ME and
oil group had good anti-nematode potential until the second hour of injury
induction, when compared to the control group (P<0.01), but no significant
differences were observed between the two groups until twenty-fourth hour
post-injury. In the histological analyzes, greater wear was observed in the oil
group muscle tissue, with an intense presence of cellular infiltrate (edema) and
muscle fibers involvement, and the same intensity of injury was not observed in
the ME group. Thus, it is concluded that pure bullfrog oil and microemulsion
system present good anti-inflammatory potential in the evaluated models,
although the pure oil showed high hepatotoxic potential, characterizing that in a
microemulsified system, it is shown with a possible new drug delivery system
(NSLF).

The benign enlargement of the prostate, better known as benign prostatic
hyperplasia (BPH) results in symptoms of the lower urinary tract (LUTS) that
contributes to lower quality of life of men over the fourth decade, and is mostly
treated by allopathic drugs of the group of 5-alpha-reductase inhibitors and / or
alpha-blockers, and / or phytotherapics, as Serenoa repens the most used. The
scientific literature shows lack research from randomised and doubleblind clinical
trials on the efficacy of homeopathic medicinal products that already have
homeopathic medical materia and new homeopathic medicinal for BPH.
The present study aimed to evaluate the effectiveness of the homeopathic product
containing Momordica charantia 12CH in the LUTS control caused by BPH in a
clinical, randomized, double-blind and placebo-controlled trial.A total of 81 patients
were randomized to group A - Placebo and group B - Momordica charantia 12CH
and evaluated at each visit by anamnesis with the International Prostate Symptom
Score (IPSS), by imaging tests measured prostate volume (PV) and post-void
residual (PVR) and laboratory tests for prostate specific antigen (PSA), quantitative
C-reactive protein (CRP) and interleukin-6 (IL-6), and in 72 samples, lymphocyte and
subpopulation immunophenotyping were performed by flow cytometry.Both the main
variable (IPSS) and the secondary variables (PSA, PV, PVR and CRP) and
accessory variables (IL-6, total lymphocytes, B lymphocytes, T lymphocytes, T helper
cells, cytotoxic T lymphocytes and CD4 / CD8 ratio and Natural Killer cells) did not
present statistical significance in the means between the groups treated and
Placebo.About the safety variables, the use of homeopathic medication by the
patients did not interfere during the six months of research, either in the fasting
glucose level, as well as in the dosages of alanine aminotransferase, aspartate
aminotransferase, alkaline phosphatase, total bilirubins and fractions and creatinine
levels used for hepatic evaluation and renal function respectively.The absence of
urinary infection confirmed by urinalysis of patients predominated in both groups A
and B throughout the clinical trial period. Clinical studies with Momordica charantia in other dynamizations to evaluate their effectiveness in the control of LUTS/BPH
should be performed, since in 12CH their action has not been proven

Toxoplasma gondii é um protozoário intracelular obrigatório amplamente distribuído, ele é o causador da toxoplasmose, doença de curso normalmente benigno que pode ter sérias repercursões em imunossuprimidos e fetos congenitamente infectados. Esta doença é uma das principais  responsáveis  por  hospitalização relacionada à alimentos e, considerando o padrão atípico pelo qual esta se observa no Brasil, os efeitos adversos no tratamento-padrão e os  fenótipos emergentes de resistência o presente estudo teve como objetivo padronizar a manutenção e determinar o perfil de patogenicidade e resistência à  sulfadiazina  de isolados locais do  T. gondii; verificar o potencial imunogênico e alterações comportamentais;  além de avaliar o potencial antiparasitário do  timol e estragol versus cepa clonal e isolados locais do parasito. O efeito citopático induzido pelo parasito foi avaliado em cultura de células RAW  264.7 e a patogenicidade foi determinada em modelo murino  após infecção com monitoramento por 30 dias e confirmada por PCR-RFLP, usando o marcador CS3. A resistência fenotípica à  sulfadiazina  foi medida utilizando de uma curva de doses em animais infectados (100, 200 ou 300 mg/kg) e a atividade anti-Toxoplasma do  timol  e  estragol  contra isolados locais e  cepa  padrão  foram  determinadas  em camundongos  Suiços  infectados e tratados durante 6 dias.  As alterações na memória/aprendizado e impulsividade foram avaliados em labirinto em cruz elevado. Polimorfismos foram determinados por sequenciamento do gene DHPS. Amostras de sangue foram coletadas de camundongos C57BL/6 para o ELISA e dosagem de citocinas em camundongos C57BL/6. O efeito citopático e a PCR-RFLP mostraram que os isolados de galinhas  representam diferentes populações do parasito. O isolado Ck3 foi o mais patogênico in vivo, o isolado moderadamente patogênico Ck2 induziu uma resposta humoral e alterações comportamentais diferentes da cepa ME49. Falhas terapêuticas foram observadas nos animais infectados com Ck3 e Pg1 e tratados com  sulfadiazina, porém não foram observados polimorfismos no gene DHPS.  Foi possível estabelecer um modelo para  determinação da  atividade  anti-Toxoplasma de novos compostos e, através deste, foi obervado que timol e estragol apresentaram efetividade contra a cepa ME49, mas apenas o  estragol apresentou ação contra o isolado Ck2,  com  forte resposta Th1.  Timol,  estragol  e  sulfadiazina não tiveram ação contra Ck3 e Pg1.  Estes perfis atípicos de patogenicidade eresistência dos parasitos locais foram observados de forma inédita e podem explicar os perfis diferenciados da toxoplasmose observados nessa região.

Os acidentes ocasionados por animais peçonhentos, pela frequência com que
ocorrem e pela mortalidade que ocasionam, representam um sério problema de saúde
pública em diversas regiões do Brasil, bem como em outros países do mundo. O uso de
extratos de plantas medicinais  como antídoto para casos de envenenamento é uma
antiga prática utilizada, ainda hoje, em muitas comunidades que não têm acesso à
soroterapia. As  plantas medicinais representam uma importante fonte de obtenção de
compostos bioativos capazes de auxiliar diretamente no tratamento do envenenamento
ou indiretamente, suplementando a soroterapia utilizada atualmente. O objetivo deste
trabalho  é  avaliar o efeito dos extratos  aquosos, frações e compostos  isolados das
espécies  Mimosa.  Tenuiflora  (jurema preta)  e  Harconia  speciosa  (mangaba)  no
processo inflamatório induzido por carragenina e as peçonhas de Bothrops  jararaca  e
Tityus. serrulatus. Os resultados demonstraram que tanto  os extratos de M. tenuiflora
quanto  de  H. speciosa  foram capazes de inibir a migração celular e a produção de
citocinas no modelo de peritonite induzido por carragenina e peçonha de T. serrulatus.
No modelo de envenenamento por  B. jararaca, os camundongos  tratados com  os
extratos das plantas em estudo reduziram o influxo leucocitário para a cavidade
peritoneal. Por último, os extratos das plantas M. tenuiflora e H. speciosa apresentaram
atividade antiflogística, reduzindo a formação do edema exercendo também uma ação
inibitória da migração de leucócitos e dano tecidual na inflamação local induzida pela
peçonha de B. jararaca. Além disso, foi desenvolvida a análise fitoquímica de ambos os
extratos aquosos obtidos. Nessa abordagem,foi possível identificar, por meio de
Cromatografia em Camada Delgada  (CCD), a presença de flavonóides e saponinas ou
terpenos no extrato aquoso da espécie vegetal M. tenuiflora. Paralelamente, por meio da
análise por Cromatografia Líquida de Alta Eficiência (CLAE), foi possível identificar a
presença de rutina e ácido clorogênico no extrato aquoso da planta H. speciosa. A partir
dos resultados apresentados, pode-se concluir então, que a administração dos extratos,
frações e compostos isolados  de M. tenuiflora  e H. speciosa  resultou na inibição do
processo inflamatório em diversos modelos experimentais. Este estudo demonstra, pela
primeira vez, o efeito das plantas M. tenuiflora e H. speciosa na inibição da inflamação
causada pelas peçonhas de B. jararaca e T. serrulatus.

Durante as últimas décadas têm sido demonstrado que existe uma relação entre a depressão maior e a ativação do sistema imunológico. Nociceptina/orfanina FQ (N/OFQ) é o ligante natural do receptor acoplado à proteína Gi chamado NOP, ambos constituem um sistema peptídico que está envolvido na regulação do humor e de respostas inflamatórias. Considerando essas ações,  a presente tese  teve como objetivo investigar as consequências do bloqueio  da  sinalização  do receptor  NOP  nos comportamentos doentio e do tipo depressivo induzidos pela administração de lipopolissacarídeo (LPS) em camundongos. A administração sistêmica de doses de LPS, que não causam  sepse, induzem alterações nos  comportamentos  de camundongos relacionadas com a atividade das citocinas pró-inflamatórias fator de necrose tumoral-α (TNF-α) e interleucinas 6 (IL-6) e 1β  (IL-  1  β). Após  2 a 6 h e 24 h  da injeção intraperitoneal,  camundongos  tratados com LPS  apresentam, respectivamente,  o comportamento doentio e o comportamento do tipo depressivo. No presente trabalho, a administração de LPS (0,8 mg/kg, ip) induziu sinais de doença em camundongos Swiss e CD-1, como perda de peso, redução transitória da temperatura retal e diminuição da ingestão de  ração e água. Além disso, nas 24 h após a injeção de LPS, essas mesmas linhagens de camundongos mostraram aumento no tempo de imobilidade durante os 6 min que foram submetidos ao teste de suspensão pela cauda (TSC). O tratamento com nortriptilina (30 mg/kg,  ip, 60 minutos antes do TSC) reduziu o tempo de imobilidade dos  camundongos  controle  e  tratados com LPS,  e foi utilizado como antidepressivo padrão. A administração prévia ao LPS do antagonista do receptor NOP SB-612111 (10 mg/kg,  ip), não alterou os comportamento doentio e do tipo depressivo induzidos pelatoxina. No entanto, quando injetado 24 h após o tratamento com LPS, SB-612111 (ip, 30 minutos antes do TSC), como também  o antagonista  peptídico  do receptor NOP UFP-101 (10 nmol/2ul,  icv, 5 min antes do TSC),  inverteram  significativamente os efeitos  da toxina. O protocolo de  indução do comportamento do tipo depressivo pela administração intraperitoneal de LPS também foi testado  em  camundongos nocautes para o receptor NOP (NOP(-/-)) e seus respectivos wild types (NOP(+/+)). O tratamento com LPS provocou alterações significativas, como a redução temporária da temperatura retal  nos camundongos NOP(-/-) e  perda de peso corporal e redução no consumo de ração e água em ambos os camundongos NOP(+/+) e NOP(-/-). O consumo de água foi significativamente diferente  entre os  genótipos. A injeção de LPS  induziu alterações transitórias nas citocinas pró-inflamatórias. Nas 6 horas após o tratamento com LPS, os níveis séricos de TNF-α mostraram-se aumentados significativamente nos camundongos NOP (+/+) e NOP (-/-),  já  os níveis de IL-6  mostraram-se  significativamente aumentados apenas no soro dos camundongos NOP (+/+). Nas 24 horas após a injeção de LPS, os níveis séricos das citocinas pró-inflamatórias retornaram à linha de base. O tratamento com LPS provocou efeitos de tipo depressivo nos camundongos NOP (+/+), mas foi ineficaz nos camundongos NOP (-/-). Os dados obtidos nesta tese mostram que o bloqueio farmacológico  e genético  da sinalização mediada pelo receptor NOP não previne os comportamentos doentio e do tipo depressivo induzidos por LPS, no entanto revertem o comportamento do tipo depressivo.  Em conclusão, esses resultados evidenciam o envolvimento do sistema peptídico N/OFQ - receptor NOP na modulação dos comportamentos relacionados ao humor e à ativação do sistema imunológico.

O topiramato é um agente antiepiléptico que tem se mostrado promissor no tratamento da enxaqueca, controle da obesidade e em diversos transtornos psiquiátricos, incluindo o transtorno bipolar. Estudos clínicos apontam o uso do topiramato para alívio da impulsividade comportamental associada ao consumo abusivo, fissura e sintomas psíquicos decorrentes da retirada do etanol. Considerando o perfil farmacocinético, farmacodinâmico e a alta tolerabilidade do topiramato na clínica comparada aos ansiolíticos e antidepressivos disponíveis, é de grande interesse investigar o potencial ansiolítico e antidepressivo deste fármaco em ratos  naive  e abstinentes ao etanol. No presente estudo foi  investigado se o tratamento agudo e crônico com topiramato altera comportamentos relacionados à ansiedade, à depressão, e os decorrentes da retirada do etanol após consumo crônico forçado. Ratos Wistar adultos machos foram tratados com topiramato (10 e 40 mg/Kg/mL) ou salina, por gavagem, 60 minutos antes dos testes do labirinto em cruz elevado (LCE) e campo aberto (CA) e natação forçada (TNF). Observou-se que tanto  a administração aguda quanto a administração crônica do topiramato (40 mg/Kg/mL) promoveu ação ansiolítica em animais naive e ação antidepressiva, após tratamento agudo, sem alteração da atividade locomotora.  Em outra série experimental, ratos foram submetidos a concentrações crescentes de etanol (2%, 4% e 6%) durante 21 dias, como única fonte de dieta líquida, enquanto que o grupo controle recebeu água  ad libitum. Após a exposição crônica, o etanol foi substituído por água, e os animais foram avaliados no LCE, CA, rota-rod, reconhecimento de objetos e TNF.  Os ratos expostos cronicamente ao etanol  exibiram comportamento ansiogênico, 72 h após a retirada da droga. Além disso, déficits de memória e comportamento do tipo depressivo, sem alteração na atividade locomotora e coordenação motora, foram observados em ratos abstinentes em longo prazo. Em uma terceira etapa deste trabalho, foi avaliado o efeito da administração aguda e crônica de topiramato  (40 mg/Kg/mL) no LCE, CA e NF em ratos abstinentes (a curto e longo prazo) ou não abstinentes a exposição forçada com etanol por 21 dias. A administração aguda e crônica com topiramato (40 mg/Kg/mL) reverteu o comportamento ansiogênico de ratos abstinentes em curto e longo prazo, sem afetar a locomoção. No entanto, a administração crônica de topiramato não contrabalanceou o comportamento do tipo depressivo de ratos abstinentes ao etanol.  O topiramato induziu um efeito ansiolítico após a administração aguda e crônica em ratos  naive  e abstinentes ao etanol. Também foi visto que, a administração aguda do topiramato provoca efeitos antidepressivos em ratos naive, porém este efeito se dissipa após tratamento crônico. Além disso, o topiramato não foi bem sucedido em aliviar o comportamento tipo depressivo em ratos abstinência. Contudo,  topiramato parece ser uma opção terapêutica interessante para o tratamento da ansiedade, inclusive aquela provocada pela retirada do etanol.

The availability of pediatric formulation for treating tuberculosis is a reality for the
countries of Asia and Africa, but not in other countries that have considerable incidence of this
disease. Considering the situation of the global need for medicines for pediatric use and
specifically for this type of product for the treatment of tuberculosis, whose drugs association
represents a major challenge, this study aimed to develop oral formulations containing
rifampicin, isoniazid and pyrazinamide in fixed-dose combination for pediatric use. It began
with the physico-chemical characterization of active ingredients (identification by IR, UV and
HPLC, determination of content by HPLC and UV, bulk density and angle of repose). It then
carried the evaluation of thermal compatibility of active ingredients with different excipients
(DSC and TG); the vehicle development for use as a carrier of oral drugs in children with
minimal components, at low concentrations, selecting them from the inputs considered safe for
children (evaluating viscosity, pH, palatability, physical and microbiological stability). It was
developed and validated spectrophotometric analytical methodology for use in the assay and
dissolution profile of three drugs in the developed products, in addition to pre-formulation
studies by apparent density and angle of repose of the individual drugs and their mixtures with
aerosil, granulation with different polymers and their effect on the release of rifampicin,
rifampicin stability in function of pH, coating and microencapsulation of rifampicin with
chitosan, and finally the development of solid oral formulations in the form of powder for
reconstitution and dispersible tablet containing a fixed-dose combination of rifampicin,
isoniazid and pyrazinamide. We evaluated the viscosity, pH, content of drug and stability of the
reconstituted preparations, as well as average weight, hardness, disintegration time, dissolution
profile and friability of the tablets. The Ozawa method was used for DSC studies kinetic to
estimate the frequency factor, the activation energy and the order of reactions that lead to
degradation of solid dosage forms. The two solid dosage forms when dispersed in water,
resulting in pseudo-plastic suspension easy to be handled, measured and administered, and can
be used for up to 12 hours after reconstitution.

Avaliação da genotoxicidade em mulheres com Síndrome dos Ovários Policísticos:

Impacto da dieta

Introdução:  Síndrome dos Ovários Policísticos (SOP), presente em 6-12% das mulheres em idade reprodutiva, é caracterizada  pelo  hiperandrogenismo,  resistência à insulina (RI) e por seu estado inflamatório, fatores exacerbados pela  obesidade  e associados  com o  aumento no  dano  de DNA. A  perda de peso,  aliada  à alimentação saudável, atua restabelecendo as funções reprodutivas e metabólicas na SOP, entretanto sua influencia na redução  do  dano de DNA,  na SOP,  são  desconhecidos. Objetivos: investigar se existem diferenças entre os marcadores de dano de DNA em mulheres com SOP e controle,  e avaliar  a efetividade da  intervenção nutricional  nos marcadores de dano de DNA e nos marcadores de risco cardiometabólicos em mulheres sobrepeso e obesas com SOP. Metodologia: as participantes tinham faixa etária entre 18 e 35 anos. No primeiro  estudo,  caso-controle prospectivo,  foram incluídas  27  mulheres  com diagnóstico  de SOP  e 20 controles. O segundo estudo foi um ensaio clínico  de intervenção nutricional com duração de 12 semanas com dieta de restrição calórica de 500Kcal/dia. A genotoxicidade, dano de DNA (tail intensity, tail moment e tail length), foi avaliada pelo  teste do cometa. Dados  antropométricos,  de consumo alimentar, hormonais, bioquímicos e inflamatórios foram avaliados nos distintos estudos. Resultados: não houve diferença significativa entre o marcador de dano de DNA  tail intensity na SOP  (24± 6,32% vs 21,7± 5,22% ) (p= 0,18) quando comparado ao grupo controle, assim como nos marcadores de tail moment (p= 0,76) e tail length ( p=0,109). Dados após  intervenção nutricional,  em  mulheres  SOP  com  sobrepeso e obesidade exibiram uma diminuição dos marcadores de danos de DNA: tail intensity (24,35 ± 5,86 – pré-dieta vs. 17,15 ± 5,04 -pos-dieta) e tail moment (20,47 ± 7,85 – pré-dieta vs. 14,13 ± 6,29  -pós-dieta) (p <0,001). Redução da ingestão de calorias, perda de peso  (3,5%), diminuição do hormônio sexual e marcadores cardiometabólicos, tais como insulina, HOMA-IR e colesterol LDL foram também reduzidos. Conclusão: mulheres com SOP não apresentam aumento na genotoxicidade identificadas pelos danos de DNA, quando comparadas a mulheres sem SOP. A intervenção nutricional reduziu a genotoxicidade de mulheres sobrepeso e obesas  com SOP, além de reduzir os fatores de  riscos cardiometabólicos.

A asma é uma doença crônica caracterizada por dispneia, tosse, espirro intermitente e opressão torácica, sintomas decorrentes de processos fisiológicos como edema, aumento de secreção de muco e contração da musculatura lisa brônquica e também encontra nas plantas medicinais sugestões para seu tratamento. A  espécie Cissampelos sympodialis Eichl é uma espécie  vegetal  bastante  estudada,  tendo sido  avaliadas e comprovadas uma série de ações farmacológicas (anti-anafilática,  anti-inflamatória  e efeito imunomodulador) que a colocam em posição de destaque para terapia da asma. Esses efeitos farmacológicos são associados à presença de alcaloides,  destacando-se a warifteína.  O controle de qualidade do material vegetal segundo métodos farmacopeicos foi realizado na busca de especificações de qualidade para as folhas de Cissampelos sympodialis e garantia de uso seguro desse insumo vegetal. A aplicação de técnicas quimiométricas do tipo planejamento experimental foi útil no estabelecimento de condições ótimas de extração para obtenção de extratos hidroalcoólcios das folhas de Cissampelos sympodialis, cuja otimização ocorreu também através de análise univariada relacionada ao tamanho de partícula do material vegetal e método de extração. A monitoração de atividade biológica anti-inflamatória, presumida através de modelos de cultura de células de linfonodos e macrófagos para quantificação de citocinas associadas ao processo inflamatório,  foi realizada para os extratos com maior e menor teor de warifteína, decorrentes do planejamento experimental, na forma de suas respectivas  frações aquosas. A relação do teor de warifteína com a potência de atividade anti-inflamatória pôde ser sugerida, havendo ainda necessidade de otimização da obtenção da fração aquosa. As condições padronizadas para obtenção do extrato hidroalcoólico envolvem a utilização de material vegetal com partículas de 500 µm, na proporção 1:10 (droga: solvente p/v), com o sistema de solvente extrator sendo etanol : água na proporção 80 : 20 (v/v),  através da maceração por 48h, com reposição de solvente após as primeiras 24 h. Foram realizados alguns ensaios de secagem por  spray dryer  das frações aquosas obtidas,  sendo sugerida a utilização de maltodextrina, isolada ou em associação a dióxido de silício coloidal, havendo ainda necessidade de ensaios definitivos. Todo o estudo foi realizado  com o intuito de se obter insumo  farmacêutico estável e de qualidade para futuro desenvolvimento de formulações e consequente obtenção de medicamento fitoterápico para o tratamento de asma.

O trissoralen (Tri) é um fármaco de baixa dosagem utilizado no tratamento da psoríase e outras dermatoses. O objetivo do trabalho foi aplicação da análise térmica e técnicas complementares para caracterização, avaliação da estabilidade do trissoralen e dos componentes das formulações farmacêuticas manipuladas. O comportamento térmico do Tri por TG/DTG-DTA em atmosfera dinâmica de ar sintético e nitrogênio evidenciou o mesmo perfil com um pico de fusão seguido de um evento relacionado à volatilização. A partir das curvas TG/DTG observou uma única etapa de perda de massa. Ao aquecer o fármaco na estufa nas temperaturas de 80, 240 e 260 °C observou-se que o mesmo não apr esentou alterações na sua estrutura química através das técnicas de DRX, CLAE, MIR, MO e MEV. A partir dos estudos cinéticos não-isotérmicos e isotérmicos por TG foi possível calcular a energia de ativação e ordem de reação para o Tri. O fármaco apresentou uma boa estabilidade térmica. Estudos de compatibilidade fármaco-excipiente mostraram que há interação do trissoralen com lauril sulfato sódio na proporção 1:1. Não se observou interação com aerosil, amido pré-gelatinizado, amido glicolato sódico, celulose, croscarmelose, estearato de magnésio, lactose e manitol. A caracterização de três formulações de trissoralen nas concentrações de 2,5; 5; 7,5; 10; 12,5 e 15 mg foi realizada por DSC, TG/DTG, DRX, MIR e NIR. Um método de classificação PCA baseado nos dados dos espectros MIR e NIR das formulações de trissoralen permitiu uma diferenciação bem sucedida em três grupos. A formulação 3 foi a que melhor apresentou o perfil analítico com a seguinte composição de excipientes aerosil, amido pré-gelatinizado e celulose. A energia de ativação do processo de volatização do fármaco foi determinada nas misturas binárias e formulação 3 por meio dos métodos isoconversional e  fitting. A mistura binária com amido glicolato sódico e lactose apresentaram diferença nos parâmetros cinéticos em comparação ao fármaco isolado. As técnicas termoanalíticas (DSC e TG/DTG) mostraram ser metodologias promissoras para quantificação do trissoralen pela linearidade obtida, seletividade, não uso de solventes, sem preparo de amostra, rapidez e praticidade.

Esse estudo propôs aplicar as técnicas térmicas, calorimetria exploratória diferencial (DSC), análise térmica diferencial (DTA) e termogravimetria/ termogravimetria derivada (TG/DTG) e não-térmicas,  espectroscopia no infravermelho (IV) e difração de raios-X (DRX) para caracterizar as propriedades físico-químicas dos fármacos despigmentantes, hidroquinona (HQ), ácido retinóico (RET), ácido kójico (KOJ), ácido glicólico (GLICOL), ácido fítico (FIT) e ácido glicirrízico (GLICIR) e dos excipientes usados em formas farmacêuticas semissólidas, bem como avaliar as possíveis interações fármaco/fármaco e fármaco/excipiente a temperatura ambiente e em condições forçadas de temperatura de exposição. Os excipientes analisados foram álcool cetílico (ACETI), álcool cetoestearílico e cetilestearilsulfato de sódio (ACETO), metilparabeno (MTP), propilparabeno (PPP), dipropilenoglicol (DPG), glicerina (GLI), EDTA dissódico (EDTA), hidroxipropilmetilcelulose (HPMC), oleato de decila (ODC) e imidazolidinil uréia (IMD). Os fármacos HQ, RET e KOJ apresentaram um intenso evento endotérmico relacionado com sua fusão, porém não foi possível identificar a fusão dos fármacos GLICOL, FIT e GLICIR, a partir das curvas DSC e DTA. A partir das curvas TG/DTG observaram-se uma única etapa de perda de massa para HQ e RET, sendo duas para o KOJ, três para o GLICOL e quatro etapas para o FIT e o GLICIR. O aquecimento dos fármacos isolados, HQ, RET e KOJ, revelou uma estabilidade química e cristalina pela manutenção, respectivamente, dos grupos funcionais e dos picos de difração dos fármacos, exceto pela modificação da cristalinidade do RET com o aquecimento. Nos estudos de compatibilidade fármaco-fármaco, observaram-se interações físicas entre HQ e KOJ, HQ e GLICIR, RET e KOJ, RET e GLICOL, RET e FIT a partir das curvas térmicas, bem como a técnica do IV mostrou interação química a temperatura ambiente entre HQ e GLICOL e RET e GLICIR. Nos estudos de compatibilidade fármaco-excipiente, observaram-se interações físicas a partir das curvas térmicas da HQ com MTP, PPP, DPG, HPMC e IMD; do RET com ACETI, ACETO, MTP e PPP; e do KOJ com MTP, PPP, GLI e IMD. Foram observadas interações químicas a temperatura ambiente da HQ com EDTA, do RET com IMD e do KOJ com IMD, notadamente destacada pelo desaparecimento de bandas de absorção importantes desses fármacos quando em mistura. O aquecimento das misturas binárias, tanto no estudo fármaco-fármaco como fármaco-excipiente, favoreceu em alguns casos o surgimento de interações líquido-líquido entre os componentes estudados, observadas pelos espectros IV e difratogramas, favorecendo o aumento das interações inter e intramoleculares. Dessa maneira, não são recomendadas em uma mesma preparação farmacêutica as associações dos fármacos HQ com KOJ, HQ com GLICOL, HQ com GLICIR, RET com KOJ, RET com GLICOL, RET com FIT e RET com GLICIR, podendo ser utilizados separadamente. Recomenda-se evitar os excipientes DPG, EDTA, HPMC e IMD em formulações com HQ, bem como o ACETI, ACETO, MTP, PPP e IMD em formulações com RET em sua composição. Sugere-se evitar MTP, PPP e IMD em formulações com KOJ. Assim, propõe-se realizar outros estudos para avaliar a compatibilidade desses fármacos com outros excipientes utilizados na manipulação magistral como substitutos daqueles que demostraram incompatibilidade com a HQ, RET e KOJ. 

O alfa-ácido lipóico (AAL) é um potente antioxidante, com favoráveis efeitos
antiinflamatórios, metabólicos e endoteliais, assim intensamente  investigado no combate aos
fatores de risco cardiovascular. O estudo teve como objetivo avaliar o efeito da
suplementação oral do AAL sobre biomarcadores de estresse oxidativo, inflamação e fatores
de risco cardiovascular em portadores de hipertensão. Trata-se de um estudo clínico
randomizado, duplo-cego e controlado com placebo, onde a intervenção foi avaliada
prospectivamente, comparando os resultados nos dois grupos. A amostra foi constituída por
64 indivíduos hipertensos que foram distribuídos aleatoriamente em grupo ALA (n=32), que
recebeu 600 mg/dia de AAL  por doze semanas e grupo controle (n=32), que recebeu o
placebo pelo mesmo período. Foram avaliados antes e depois da intervenção: substâncias
reativas ao ácido tiobarbitúrico (SRAT), conteúdo da glutationa reduzida (GSH); atividades
enzimáticas da glutationa peroxidase (GPx) e da superóxido dismustase; proteína C reativa
(PCR-us); triglicerídeos, colesterol total e frações; glicemia de jejum; e indicadores
antropométricos. Observou-se uma redução estatisticamente significativa (p < 0.05) nas
concentrações séricas de colesterol total, lipoproteína de muita baixa densidade (VLDL),
lipoproteína de alta densidade (HDL), triglicerídeos e glicose. Também houve redução do
peso corporal e das circunferências abdominais, da cintura e do quadril no grupo que recebeu
o AAL. Ainda, observou-se um aumento estatisticamente significativo (p < 0.05) do conteudo
de glutationa reduzida (GSH) e da atividade da glutationa peroxidase  (GPx) no grupo que
recebeu o AAL.  A  administração oral do AAL  mostra-se como um valioso adjuvante
terapêutico, que pode contribuir com a diminuição dos danos causados pelo estresse oxidativo
e outros fatores de risco que estão associados com o processo aterosclerótico.