Background:: Although several research efforts have been made worldwide to discover novel drug ca... more Background:: Although several research efforts have been made worldwide to discover novel drug candidates for the treatment of Chagas disease, the nitroimidazole drug benznidazol remains the only therapeutic alternative in the control of this disease. However, this drug presents reduced efficacy in the chronic form of the disease and limited safety after long periods of administration, making it necessary to search for new, more potent and safe prototypes. Objective:: We described herein the synthesis and the trypanocidalaction of new functionalized carbohydrazonamides (2-10) against trypomastigote forms of Trypanosoma cruzi. Methods: These compounds were designed through the application of molecular hybridization concept between two potent anti-T. cruzi prototypes, the nitroimidazole derivative megazol (1) and the cinnamyl N-acylhydrazone derivative (14) which have been shown to be twice as potent in vitro as benznidazole. Results:: The most active compounds were the (Z)-N'-((E)-3-(4-nitrophenyl)-acryloyl)-1-methyl-5- nitro-1H-imidazol-2-carbohydrazonamide (6) (IC50=9.50 μM) and the (Z)-N'-((E)-3-(4- hydroxyphe-nyl)-acryloyl)-1-methyl-5-nitro-1H-imidazol-2-carbohydrazonamide (8) (IC50=12.85 μM), which were almost equipotent to benznidazole (IC50=10.26 μM) used as standard drug. The removal of the amine group attached to the imine subunit in the corresponding N-acylhydrazone derivatives (11-13) resulted in less potent or inactive compounds. The para-hydroxyphenyl derivative (8) presented also a good selectivity index (SI = 32.94) when tested against mammalian cells from Swiss mice. Conclusion:: The promising trypanocidal profile of new carbohydrazonamide derivatives (6) and (8) was characterized. These compounds have proved to be a good starting point for the design of more effective trypanocidal drug candidates.
Mitogen-activated protein kinase (MAPK) signaling is strongly implicated in cardiovascular remode... more Mitogen-activated protein kinase (MAPK) signaling is strongly implicated in cardiovascular remodeling in pulmonary hypertension (PH) and right ventricle (RV) failure. The effects of a newly designed p38 inhibitor, LASSBio-1824, were investigated in experimentally induced PH. Male Wistar rats were exposed to hypoxia and SU5416 (SuHx), and normoxic rats were used as controls. Oral treatment was performed for 14 days with either vehicle or LASSBio-1824 (50 mg/kg). Pulmonary vascular resistance and RV structure and function were assessed by echocardiography and catheterization. Histological, immunohistochemical and Western blot analysis of lung and RV were performed to investigate cardiovascular remodeling and inflammation. Treatment with LASSBio-1824 normalized vascular resistance by attenuating vessel muscularization and endothelial dysfunction. In the heart, treatment decreased RV systolic pressure, hypertrophy and collagen content, improving cardiac function. Protein content of TNF-...
Background: Prostate cancer is the second most frequently diagnosed malignancy worldwide. Here, t... more Background: Prostate cancer is the second most frequently diagnosed malignancy worldwide. Here, the cytotoxic and antimetastatic effects of a new HDAC6/8 inhibitor, LASSBio-1911, and a new dual-PI3K/HDAC6 inhibitor, LASSBio-2208, were evaluated against PC3 prostate cancer cell line. Methods: A MTT assay was used to assess the cell viability. Annexin V/propidium iodide (PI) was used to detect apoptotic cell death and to analyze the cell cycle distribution. Interleukin 6 (IL-6) levels were measured by ELISA. A cell scratch assay was performed to assess cell migration, and the expression of proteins was estimated by Western blotting. Results: LASSBio-1911 and LASSBio-2208 exert cytotoxic effects against PC3 cells. However, LASSBio-2208 was demonstrated to be more potent than LASSBio-1911. The apoptosis assays showed that both compounds trigger apoptotic processes and cause the arrest of cells in the G2/M phase of the cell cycle. The Western blot analysis revealed that LASSBio-2208 sign...
The deregulation of the Wnt/β-catenin signaling pathway is a central event in colorectal cancer p... more The deregulation of the Wnt/β-catenin signaling pathway is a central event in colorectal cancer progression, thus a promising target for drug development. Many natural compounds, such as flavonoids, have been described as Wnt/β-catenin inhibitors and consequently modulate important biological processes like inflammation, redox balance, cancer promotion and progress, as well as cancer cell death. In this context, we identified the chalcone lonchocarpin isolated from Lonchocarpus sericeus as a Wnt/β-catenin pathway inhibitor, both in vitro and in vivo. Lonchocarpin impairs β-catenin nuclear localization and also inhibits the constitutively active form of TCF4, dnTCF4-VP16. Xenopus laevis embryology assays suggest that lonchocarpin acts at the transcriptional level. Additionally, we described lonchocarpin inhibitory effects on cell migration and cell proliferation on HCT116, SW480, and DLD-1 colorectal cancer cell lines, without any detectable effects on the non-tumoral intestinal cell...
Tumor aggressiveness is usually associated with metastasis. MDA-MB 231, a triple-negative breast ... more Tumor aggressiveness is usually associated with metastasis. MDA-MB 231, a triple-negative breast cancer (TNBC), is an aggressive type of breast cancer and associated with early metastasis. The Rho/ROCK pathway is a key regulator of cell motility involving cytoskeleton regulation through stabilization of actin filaments and stress fiber formation. In this study we show that Fasudil, a ROCK inhibitor, inhibited the migration of MDA-MB 231 and A549 cells, without altering the viability of these cells at the concentration of 10 μM, modified tumor cell morphology, with disorganization of stress fibers and promotes activation of the canonical-Wnt/beta-catenin pathway. Therefore, Fasudil present a promising approach to the prevention of breast cancer metastasis through a different mechanism of action from the well-known one.
The Journal of pharmacology and experimental therapeutics, Aug 18, 2016
Rheumatoid arthritis is an inflammatory autoimmune condition and tumor necrosis factor-α (TNF-α) ... more Rheumatoid arthritis is an inflammatory autoimmune condition and tumor necrosis factor-α (TNF-α) plays an important role in its pathophysiology. In vitro, (E)-N'-(3,4-dimethoxybenzylidene)-N-methylbenzohydrazide (LASSBio-1359) has exhibited anti-TNF-α properties and in vivo these effects are mediated via activation of adenosine receptor. This work investigates the antinociceptive action of LASSBio-1359 in murine models of acute and chronic inflammatory pain. Male mice received an intraperitoneal injection of LASSBio-1359 and then were evaluated in formalin- and carrageenan-induced paw edema assays. Complete Freund's Adjuvant (CFA) was used to induce a mouse model of monoarthritis. These mice were treated with LASSBio-1359 by oral gavage to evaluate thermal and mechanical hyperalgesia. TNF-α and i-NOS expression, as well as histological features were also analyzed. The time of reactivity to formalin in the neurogenic phase was reduced from 56.3 ± 6.0 s to 32.7 ± 2.2 s and 23....
Acta Crystallographica Section A Foundations and Advances, 2014
Hypertension is considered a multi-factorial disease being an important worldwide public-health c... more Hypertension is considered a multi-factorial disease being an important worldwide public-health challenge due to its high frequency and concomitant risks of cardiovascular and other diseases. It has been identified as the leading risk factor for mortality, and is ranked as the third cause of disability-adjusted life-years[1]. The LASSBio-1289 compound was designed from the lead compound LASSBio-294 through the use of medicinal chemistry strategy of bioisosteric replacement of aromatic rings[2]. Many factors are involved in the task of describing the relationship between chemical structure and pharmacological activity of a prototype of a new drug. Among these factors the study of atomic-level structure can be included. The aim of this work is to carry out the structural characterization of LASSBio-1289, in solid-state. Since many NAH derivatives, as LASSBio-1289, do not form single crystals for proper structure determination, this task was overcome by using X-ray powder diffraction d...
A high-performance liquid chromatographic (HPLC) and a ultraviolet (UV) methods were developed an... more A high-performance liquid chromatographic (HPLC) and a ultraviolet (UV) methods were developed and validated for the quantitative determination of Dexibuprofen (DI) in pharmaceutical dosage form. HPLC was carried out by reversed phase technique on a RP-18 column with a mobile phase composed of acetonitrile and 0.5% triethylamine (pH 7.5 adjusted with orthophosphoric acid (30 : 70, v/v)). UV method was performed with the λ max at 222.0 nm. Both the methods showed good linearity, reproducibility and precision. No spectral or chromatographic interferences from the tablet excipients were found in UV and HPLC. The method was successfully applied to commercial DEXIFEN tablets. Validation parameters such as linearity, precision, accuracy, and specificity were determined. The proposed method could be applicable for routine analysis of DI and monitoring of the quality of marketed drugs.
Background and purpose: Compound LASSBio‐881 is an orally effective antinociceptive that binds t... more Background and purpose: Compound LASSBio‐881 is an orally effective antinociceptive that binds to cannabinoid receptors and is active mainly on the neurogenic component of pain models. We investigated whether transient receptor potential vanilloid subfamily type 1 (TRPV1) channels are involved in the effects of LASSBio‐881.Experimental approach: Modulation of capsaicin (CAP)‐ and low pH‐induced currents was evaluated in TRPV1‐expressing Xenopus oocytes. In vivo effects were evaluated in CAP‐induced acute and inflammatory changes in nociception, as well as in partial sciatic ligation‐induced thermal hypernociception.Key results: LASSBio‐881 inhibited TRPV1 currents elicited by CAP with an IC50 of 14 µM, and inhibited proton‐gated currents by 70% at 20 µM. Functional interaction with CAP was surmountable. Locally applied LASSBio‐881 decreased time spent in CAP‐elicited nocifensive behaviour by 30%, and given orally it reduced measures of CAP‐ or carrageenan‐evoked thermal hypernoci...
Background: The drugs available for the treatment of Chagas disease are ineffective in its chroni... more Background: The drugs available for the treatment of Chagas disease are ineffective in its chronic phase and produce many adverse effects, making the search for new drugs an urgent medical need. Objective: This study aimed to design, synthesize, and evaluate the trypanocidal and cytotoxic profiles of new 4-nitroimidazole prototypes. Method: The new compounds were synthesized in overall yields ranging from 31-to 52% through the use of classical and reproducible methodologies. Their trypanocidal profile and cytotoxicity were assayed against trypomastigote forms of T. cruzi and mammalian macrophages, respectively. Results: The best trypanocidal activity was evidenced in compounds that present lipophilic and electronegative substituents, e.g. 4-chlorophenyl derivative (5), with an IC50 = 206.98 μM; Conclusion: The new benznidazole N-glycinyl-hydrazone analogues demonstrated a very significant reduction in the trypanocidal activity compared to benznidazole, which seems to be related to the position occupied by the nitro group in the imidazole ring.
Background:: Although several research efforts have been made worldwide to discover novel drug ca... more Background:: Although several research efforts have been made worldwide to discover novel drug candidates for the treatment of Chagas disease, the nitroimidazole drug benznidazol remains the only therapeutic alternative in the control of this disease. However, this drug presents reduced efficacy in the chronic form of the disease and limited safety after long periods of administration, making it necessary to search for new, more potent and safe prototypes. Objective:: We described herein the synthesis and the trypanocidalaction of new functionalized carbohydrazonamides (2-10) against trypomastigote forms of Trypanosoma cruzi. Methods: These compounds were designed through the application of molecular hybridization concept between two potent anti-T. cruzi prototypes, the nitroimidazole derivative megazol (1) and the cinnamyl N-acylhydrazone derivative (14) which have been shown to be twice as potent in vitro as benznidazole. Results:: The most active compounds were the (Z)-N'-((E)-3-(4-nitrophenyl)-acryloyl)-1-methyl-5- nitro-1H-imidazol-2-carbohydrazonamide (6) (IC50=9.50 μM) and the (Z)-N'-((E)-3-(4- hydroxyphe-nyl)-acryloyl)-1-methyl-5-nitro-1H-imidazol-2-carbohydrazonamide (8) (IC50=12.85 μM), which were almost equipotent to benznidazole (IC50=10.26 μM) used as standard drug. The removal of the amine group attached to the imine subunit in the corresponding N-acylhydrazone derivatives (11-13) resulted in less potent or inactive compounds. The para-hydroxyphenyl derivative (8) presented also a good selectivity index (SI = 32.94) when tested against mammalian cells from Swiss mice. Conclusion:: The promising trypanocidal profile of new carbohydrazonamide derivatives (6) and (8) was characterized. These compounds have proved to be a good starting point for the design of more effective trypanocidal drug candidates.
Mitogen-activated protein kinase (MAPK) signaling is strongly implicated in cardiovascular remode... more Mitogen-activated protein kinase (MAPK) signaling is strongly implicated in cardiovascular remodeling in pulmonary hypertension (PH) and right ventricle (RV) failure. The effects of a newly designed p38 inhibitor, LASSBio-1824, were investigated in experimentally induced PH. Male Wistar rats were exposed to hypoxia and SU5416 (SuHx), and normoxic rats were used as controls. Oral treatment was performed for 14 days with either vehicle or LASSBio-1824 (50 mg/kg). Pulmonary vascular resistance and RV structure and function were assessed by echocardiography and catheterization. Histological, immunohistochemical and Western blot analysis of lung and RV were performed to investigate cardiovascular remodeling and inflammation. Treatment with LASSBio-1824 normalized vascular resistance by attenuating vessel muscularization and endothelial dysfunction. In the heart, treatment decreased RV systolic pressure, hypertrophy and collagen content, improving cardiac function. Protein content of TNF-...
Background: Prostate cancer is the second most frequently diagnosed malignancy worldwide. Here, t... more Background: Prostate cancer is the second most frequently diagnosed malignancy worldwide. Here, the cytotoxic and antimetastatic effects of a new HDAC6/8 inhibitor, LASSBio-1911, and a new dual-PI3K/HDAC6 inhibitor, LASSBio-2208, were evaluated against PC3 prostate cancer cell line. Methods: A MTT assay was used to assess the cell viability. Annexin V/propidium iodide (PI) was used to detect apoptotic cell death and to analyze the cell cycle distribution. Interleukin 6 (IL-6) levels were measured by ELISA. A cell scratch assay was performed to assess cell migration, and the expression of proteins was estimated by Western blotting. Results: LASSBio-1911 and LASSBio-2208 exert cytotoxic effects against PC3 cells. However, LASSBio-2208 was demonstrated to be more potent than LASSBio-1911. The apoptosis assays showed that both compounds trigger apoptotic processes and cause the arrest of cells in the G2/M phase of the cell cycle. The Western blot analysis revealed that LASSBio-2208 sign...
The deregulation of the Wnt/β-catenin signaling pathway is a central event in colorectal cancer p... more The deregulation of the Wnt/β-catenin signaling pathway is a central event in colorectal cancer progression, thus a promising target for drug development. Many natural compounds, such as flavonoids, have been described as Wnt/β-catenin inhibitors and consequently modulate important biological processes like inflammation, redox balance, cancer promotion and progress, as well as cancer cell death. In this context, we identified the chalcone lonchocarpin isolated from Lonchocarpus sericeus as a Wnt/β-catenin pathway inhibitor, both in vitro and in vivo. Lonchocarpin impairs β-catenin nuclear localization and also inhibits the constitutively active form of TCF4, dnTCF4-VP16. Xenopus laevis embryology assays suggest that lonchocarpin acts at the transcriptional level. Additionally, we described lonchocarpin inhibitory effects on cell migration and cell proliferation on HCT116, SW480, and DLD-1 colorectal cancer cell lines, without any detectable effects on the non-tumoral intestinal cell...
Tumor aggressiveness is usually associated with metastasis. MDA-MB 231, a triple-negative breast ... more Tumor aggressiveness is usually associated with metastasis. MDA-MB 231, a triple-negative breast cancer (TNBC), is an aggressive type of breast cancer and associated with early metastasis. The Rho/ROCK pathway is a key regulator of cell motility involving cytoskeleton regulation through stabilization of actin filaments and stress fiber formation. In this study we show that Fasudil, a ROCK inhibitor, inhibited the migration of MDA-MB 231 and A549 cells, without altering the viability of these cells at the concentration of 10 μM, modified tumor cell morphology, with disorganization of stress fibers and promotes activation of the canonical-Wnt/beta-catenin pathway. Therefore, Fasudil present a promising approach to the prevention of breast cancer metastasis through a different mechanism of action from the well-known one.
The Journal of pharmacology and experimental therapeutics, Aug 18, 2016
Rheumatoid arthritis is an inflammatory autoimmune condition and tumor necrosis factor-α (TNF-α) ... more Rheumatoid arthritis is an inflammatory autoimmune condition and tumor necrosis factor-α (TNF-α) plays an important role in its pathophysiology. In vitro, (E)-N'-(3,4-dimethoxybenzylidene)-N-methylbenzohydrazide (LASSBio-1359) has exhibited anti-TNF-α properties and in vivo these effects are mediated via activation of adenosine receptor. This work investigates the antinociceptive action of LASSBio-1359 in murine models of acute and chronic inflammatory pain. Male mice received an intraperitoneal injection of LASSBio-1359 and then were evaluated in formalin- and carrageenan-induced paw edema assays. Complete Freund's Adjuvant (CFA) was used to induce a mouse model of monoarthritis. These mice were treated with LASSBio-1359 by oral gavage to evaluate thermal and mechanical hyperalgesia. TNF-α and i-NOS expression, as well as histological features were also analyzed. The time of reactivity to formalin in the neurogenic phase was reduced from 56.3 ± 6.0 s to 32.7 ± 2.2 s and 23....
Acta Crystallographica Section A Foundations and Advances, 2014
Hypertension is considered a multi-factorial disease being an important worldwide public-health c... more Hypertension is considered a multi-factorial disease being an important worldwide public-health challenge due to its high frequency and concomitant risks of cardiovascular and other diseases. It has been identified as the leading risk factor for mortality, and is ranked as the third cause of disability-adjusted life-years[1]. The LASSBio-1289 compound was designed from the lead compound LASSBio-294 through the use of medicinal chemistry strategy of bioisosteric replacement of aromatic rings[2]. Many factors are involved in the task of describing the relationship between chemical structure and pharmacological activity of a prototype of a new drug. Among these factors the study of atomic-level structure can be included. The aim of this work is to carry out the structural characterization of LASSBio-1289, in solid-state. Since many NAH derivatives, as LASSBio-1289, do not form single crystals for proper structure determination, this task was overcome by using X-ray powder diffraction d...
A high-performance liquid chromatographic (HPLC) and a ultraviolet (UV) methods were developed an... more A high-performance liquid chromatographic (HPLC) and a ultraviolet (UV) methods were developed and validated for the quantitative determination of Dexibuprofen (DI) in pharmaceutical dosage form. HPLC was carried out by reversed phase technique on a RP-18 column with a mobile phase composed of acetonitrile and 0.5% triethylamine (pH 7.5 adjusted with orthophosphoric acid (30 : 70, v/v)). UV method was performed with the λ max at 222.0 nm. Both the methods showed good linearity, reproducibility and precision. No spectral or chromatographic interferences from the tablet excipients were found in UV and HPLC. The method was successfully applied to commercial DEXIFEN tablets. Validation parameters such as linearity, precision, accuracy, and specificity were determined. The proposed method could be applicable for routine analysis of DI and monitoring of the quality of marketed drugs.
Background and purpose: Compound LASSBio‐881 is an orally effective antinociceptive that binds t... more Background and purpose: Compound LASSBio‐881 is an orally effective antinociceptive that binds to cannabinoid receptors and is active mainly on the neurogenic component of pain models. We investigated whether transient receptor potential vanilloid subfamily type 1 (TRPV1) channels are involved in the effects of LASSBio‐881.Experimental approach: Modulation of capsaicin (CAP)‐ and low pH‐induced currents was evaluated in TRPV1‐expressing Xenopus oocytes. In vivo effects were evaluated in CAP‐induced acute and inflammatory changes in nociception, as well as in partial sciatic ligation‐induced thermal hypernociception.Key results: LASSBio‐881 inhibited TRPV1 currents elicited by CAP with an IC50 of 14 µM, and inhibited proton‐gated currents by 70% at 20 µM. Functional interaction with CAP was surmountable. Locally applied LASSBio‐881 decreased time spent in CAP‐elicited nocifensive behaviour by 30%, and given orally it reduced measures of CAP‐ or carrageenan‐evoked thermal hypernoci...
Background: The drugs available for the treatment of Chagas disease are ineffective in its chroni... more Background: The drugs available for the treatment of Chagas disease are ineffective in its chronic phase and produce many adverse effects, making the search for new drugs an urgent medical need. Objective: This study aimed to design, synthesize, and evaluate the trypanocidal and cytotoxic profiles of new 4-nitroimidazole prototypes. Method: The new compounds were synthesized in overall yields ranging from 31-to 52% through the use of classical and reproducible methodologies. Their trypanocidal profile and cytotoxicity were assayed against trypomastigote forms of T. cruzi and mammalian macrophages, respectively. Results: The best trypanocidal activity was evidenced in compounds that present lipophilic and electronegative substituents, e.g. 4-chlorophenyl derivative (5), with an IC50 = 206.98 μM; Conclusion: The new benznidazole N-glycinyl-hydrazone analogues demonstrated a very significant reduction in the trypanocidal activity compared to benznidazole, which seems to be related to the position occupied by the nitro group in the imidazole ring.
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