RESUMO
The malaria-causing blood stage of Plasmodium falciparum requires extracellular pantothenate for proliferation. The parasite converts pantothenate into coenzyme A (CoA) via five enzymes, the first being a pantothenate kinase (PfPanK). Multiple antiplasmodial pantothenate analogues, including pantothenol and CJ-15,801, kill the parasite by targeting CoA biosynthesis/utilisation. Their mechanism of action, however, remains unknown. Here, we show that parasites pressured with pantothenol or CJ-15,801 become resistant to these analogues. Whole-genome sequencing revealed mutations in one of two putative PanK genes (Pfpank1) in each resistant line. These mutations significantly alter PfPanK activity, with two conferring a fitness cost, consistent with Pfpank1 coding for a functional PanK that is essential for normal growth. The mutants exhibit a different sensitivity profile to recently-described, potent, antiplasmodial pantothenate analogues, with one line being hypersensitive. We provide evidence consistent with different pantothenate analogue classes having different mechanisms of action: some inhibit CoA biosynthesis while others inhibit CoA-utilising enzymes.
Assuntos
Antimaláricos/farmacologia , Resistência a Medicamentos , Malária/tratamento farmacológico , Mutação , Ácido Pantotênico/análogos & derivados , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Plasmodium falciparum/efeitos dos fármacos , Animais , Coenzima A/biossíntese , Eritrócitos/parasitologia , Malária/parasitologia , Ácido Pantotênico/farmacologia , Testes de Sensibilidade Parasitária , Fosforilação , Proteínas de Protozoários/genéticaRESUMO
A fast, protecting-group-free synthesis of dihydropyridinones has been developed. Starting from commercially available aldehydes, a novel one-pot amidoallylation gave access to diene compounds in good yields. Ring-closing metathesis conditions were then employed to produce the target dihydropyridinones efficiently and in high yields.
RESUMO
Anti-virulence (AV) compounds are a promising alternative to traditional antibiotics for fighting bacterial infections. The Type Three Secretion System (T3SS) is a well-studied and attractive AV target, given that it is widespread in more than 25 species of Gram-negative bacteria, including enterohemorrhagic E. coli (EHEC), and as it is essential for host colonization by many pathogens. In this work, we designed, synthesized and tested a new series of compounds that block the functionality of the T3SS of EHEC. Affinity chromatography experiments identified the primary target of the compounds as the T3SS needle pore protein EspD, which is essential for effector protein translocation into host cells. These data were supported by mechanistic studies that determined the coiled-coil domain 1 of EspD as a key compound-binding site, thereby preventing correct assembly of the T3SS complex on the cell surface. However, binding of inhibitors to EspD or deletion of EspD itself did not result in transcriptional down-regulation of effector proteins. Instead, we found the compounds to exhibit dual-functionality by also down-regulating transcription of the entire chromosomal locus encoding the T3SS, further demonstrating their desirability and effectiveness.
Assuntos
Escherichia coli Êntero-Hemorrágica/metabolismo , Sistemas de Secreção Tipo III/antagonistas & inibidores , Sistemas de Secreção Tipo III/metabolismo , Membrana Celular/metabolismo , Regulação para Baixo , Escherichia coli Enteropatogênica/metabolismo , Proteínas de Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Humanos , Domínios Proteicos , Transporte Proteico , VirulênciaRESUMO
An efficient and selective approach for the synthesis of polyfunctionalised 3-fluoropyrroles has been developed starting from commercial aldehydes. The methodology is concise, efficient and allows for the modular and systematic assembly of polysubstituted 3-fluoropyrroles. This synthesis provides an alternative and highly convergent strategy for the generation of these chemically and biologically important units.
Assuntos
Hidrocarbonetos Fluorados/síntese química , Pirróis/síntese química , Hidrocarbonetos Fluorados/química , Estrutura Molecular , Pirróis/químicaRESUMO
The diastereoselective synthesis of fluorinated δ-lactams has been achieved through an efficient five step process. The route can tolerate a range of functionalities, and provides a quick route for the generation of new fluorinated medicinal building blocks.
Assuntos
Benzaldeídos/química , Lactamas/síntese química , Halogenação , Estrutura Molecular , EstereoisomerismoRESUMO
The total synthesis of (+)-crocacin D has been achieved in 15 steps (9 isolated intermediates) and 14% overall yield from commercially available starting materials and using (+)-crocacin C as a key intermediate. A number of simplified analogues and their biological activities are also reported.
Assuntos
Amidas/síntese química , Amidas/química , Amidas/farmacologia , Animais , Antifúngicos/farmacologia , Afídeos/efeitos dos fármacos , Herbicidas/farmacologia , Inseticidas/farmacologiaRESUMO
Osteonecrosis of the jaw (ONJ) is a relatively rare side effect after prolonged use of bisphosphonates, which are drugs used to treat bone resorption in osteoporosis and certain cancers. This study introduces a novel ONJ model in rats by combining exposure to bisphosphonates, oral surgery, and bacterial inoculation. Potential ONJ preventive effects of polyguanidine (GuaDex) or antibiotics were evaluated. The study consisted of twenty-four male Wistar rats were divided into four groups. Groups 1 to 3 were given weekly doses of i.v. Zoledronic acid (ZA), four weeks before and two weeks after an osteotomy procedure on their left mandibular first molar. Group 4 was a negative control. Streptococcus gordonii bacteria were introduced into the osteotomy pulp chamber and via the food for seven days. On day eight, the rats were given different treatments. Group 1 was given a GuaDex injection into the osteotomy socket, Group 2 was given an intramuscular (i.m.) injection of clindamycin, Group 3 (positive control) was given an i.m. injection of saline, and Group 4 was given an i.m. injection of saline. Blood samples were taken two weeks after the osteotomy procedure, after which the rats were euthanized. Bone healing, bone mineral density, histology, and blood status were analyzed. The results showed that Group 1 (GuaDex) had no ONJ, extensive ongoing bone regeneration, active healing activity, vascularization, and no presence of bacteria. Group 2 (clindamycin) showed early stages of ONJ, avascular areas, and bacteria. Group 3 showed stages of ONJ, inflammatory infiltrates, defective healing, and bacterial presence, and Group 4 had normal healing activity and no bacterial presence. Conclusion: ZA treatment and bacterial inoculation after tooth extraction inhibited bone remodeling/healing and induced ONJ characteristic lesions in the rats. Only GuaDex apparently prevented ONJ development, stimulated bone remodeling, and provided an antimicrobial effect.
Assuntos
Osteonecrose da Arcada Osseodentária Associada a Difosfonatos , Ratos Wistar , Animais , Masculino , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/prevenção & controle , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/patologia , Ratos , Difosfonatos/farmacologia , Difosfonatos/efeitos adversos , Guanidinas/farmacologia , Guanidinas/uso terapêuticoRESUMO
The enantioselective synthesis of the oxa-pinnaic acid framework has been achieved through internal asymmetric induction. The synthetic strategy pursued illustrates the adaptability of the Achmatowicz oxidative rearrangement for the synthesis of complex spirocyclic pyrans starting from tertiary alcohols.
Assuntos
Alcaloides/química , Piranos/química , Piranos/síntese química , Compostos de Espiro/química , Compostos de Espiro/síntese química , Estrutura Molecular , Piranos/farmacologia , Compostos de Espiro/farmacologiaRESUMO
Enamides, dienamides, and enynamides are important building blocks in synthetic, biological, and medicinal chemistry as well as materials science. Despite the extensive breath of their potential utility in synthetic chemistry, there is a lack of simple, high-yielding methods to deliver them efficiently and as single isomers. In this paper, we present a novel, protecting group free, efficient, and stereoselective approach to the generation of ß-halo-enamides. The methodology presented provides a robust synthetic platform from which E- or Z-enamides can be generated in good yields and with complete stereocontrol.
Assuntos
Amidas/síntese química , Amidas/química , Cristalografia por Raios X , Modelos Moleculares , Estrutura Molecular , EstereoisomerismoRESUMO
The step-economic total synthesis of (+)-crocacin C has been achieved in 20% yield from commercially available starting materials. This approach requires the isolation of only 8 intermediates and can provide a reliable supply of (+)-crocacin C for the development of new antifungal and crop protection agents.
Assuntos
Alcenos/síntese química , Amidas/síntese química , Antifúngicos/síntese química , Compostos de Boro/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , EstereoisomerismoRESUMO
Biodegradation of estrogen hormone micropollutants is a well-established approach toward their remediation. Fluorescently labeled substrates are used extensively for rapid, near-real-time analysis of biological processes and are a potential tool for studying biodegradation processes faster and more efficiently than conventional approaches. However, it is important to understand how the fluorescently tagged surrogates compare with the natural substrate in terms of chemical analysis and the intended application. We derivatized three natural estrogens with BODIPY fluorophores by azide-alkyne cycloaddition click reaction and developed an analytical workflow based on simple liquid-liquid extraction and HPLC-PDA analysis. The developed methods allow for concurrent analysis of both fluorescent and natural estrogens with comparable recovery, accuracy, and precision. We then evaluated the use of BODIPY-labeled estrogens as surrogate substrates for studying biodegradation using a model bacterium for estrogen metabolism. The developed analytical methods were successfully employed to compare the biological transformation of 17ß-estradiol (E2), with and without the BODIPY fluorescent tag. Through measuring the complete degradation of E2 and the transformation of BODIPY-estradiol to BODIPY-estrone in the presence of a co-substrate, we found that BODIPY-labeled estrogens are biologically viable surrogates for investigating biodegradation in environmental bacteria.
RESUMO
BACKGROUND: Anogenital warts are a common human papillomavirus infection. They cause emotional distress, especially when they are in the anogenital region. Cryotherapy is a first-line treatment. Previous clinical trials and case series have reported variable results with retinoids (isotretinoin) as adjuvant therapy. OBJECTIVE: To determine the safety and efficacy of low-dose oral isotretinoin as adjuvant treatment of anogenital warts. METHODS: Forty-six patients with anogenital warts were randomly assigned to isotretinoin + cryotherapy (n = 23) or only cryotherapy (n = 23). Patients were allocated via an interactive web-based randomization system. Evaluators were blinded to treatments. Isotretinoin 20 mg/daily + cryotherapy or cryotherapy were prescribed for 6 weeks. Patients were followed for 4 months. Genotyping of lesions was performed before treatment started. Dermatology Life Quality Index (DLQI) and Columbia-Suicide Severity Rating Scale (C-SSRS) were measured at the beginning and end of therapy. All patients completed the study. RESULTS: Both Groups had 50% clearance at the end of treatment. Recurrence in the combined group was not significantly lower than in the cryotherapy group (P = 0.59). Improvement was observed in the DLQI of all patients in both groups (P = 0.001). No suicidal intention was detected with the C-SSRS. Two patients (one in each group) had liver function test abnormalities after treatment. CONCLUSION: Combined therapy showed a slight not significant efficacy for anogenital warts in Hispanic patients. Low-dose isotretinoin seems to be safe even when it is used with cryotherapy on anogenital warts. TRIAL REGISTRATION: On April 25, 2019 with registration number DE19-00004, CONBIOÉTICA-19-CEI-001-20160404. Prospectively registered.
Assuntos
Condiloma Acuminado/terapia , Crioterapia , Isotretinoína/administração & dosagem , Administração Oral , Adulto , Terapia Combinada , Condiloma Acuminado/diagnóstico , Condiloma Acuminado/psicologia , Relação Dose-Resposta a Droga , Feminino , Humanos , Isotretinoína/efeitos adversos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Qualidade de Vida , Recidiva , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
Tumors of the paratesticular region are generally tumors of slow growth, with little symptomatology and, in most cases, benign in nature; in this area, a borderline serous tumor may arise hypothetically from Müllerian metaplasia of the tunica vaginalis, which is histologically identical to its ovarian counterpart. We present a 10-year-old male, with right gynecomastia and ipsilateral hydrocele, showing an enlarged right testicle with a volume of 12 ml and a left testicle with a volume of 10 ml. A right orchiectomy was performed, which presented a poorly defined tan tumor of 1.8 cm that occupied the vaginal and epididymal tunica, and infiltrates the testicular parenchyma. Histological sections revealed a cystic neoplasm, with hierarchical papillary projections, covered by one or several epithelial columnar and hobnail cells with moderate atypia and scant mitosis. Immunohistochemical reactions were performed, resulting positive for PAX-8, epithelial membrane antigen, and CK7, confirming the diagnosis of borderline serous tumor. Since the first reported case in 1986, few have been reported, the majority of these in adults with only three cases in children. In the few cases reported, the prognosis is usually favorable after surgical resection, with disease-free follow-up for up to 18 years.
RESUMO
A fast and efficient one-step approach to the synthesis of dienamides is reported. This concise methodology relies on the use of imides as reactive intermediates and allows for the preferential formation of Z,E-dienamides in good yields.
Assuntos
Amidas/síntese química , Catálise , Imidas/química , Cristalização , Cristalografia por Raios X , Ciclização , Mesilatos/química , Modelos Moleculares , Estrutura Molecular , EstereoisomerismoRESUMO
Current treatment modalities for disseminated cutaneous malignant melanoma (CMM) improve survival; however, relapses are common. A number of receptor tyrosine kinases (RTKs) including EGFR and MET have been reported to be involved in CMM metastasis and in the development of resistance to therapy, targeting the mitogen-activated protein kinase (MAPK pathway). IHC analysis showed that patients with higher MET protein expression had a significantly shorter overall survival. In addition, silencing of MET caused an upregulation of EGFR and p-AKT, which was abrogated by concomitant silencing of MET and EGFR in CMM cells resistant to MAPK-targeting drugs. We therefore explored novel treatment strategies using clinically approved drugs afatinib (ERBB family inhibitor) and crizotinib (MET inhibitor), to simultaneously block MET and ERBB family RTKs. The effects of the combination were assessed in cell culture and spheroid models using established CMM and patient-derived short-term cell lines, and an in vivo xenograft mouse model. The combination had a synergistic effect, promoting cell death, concomitant with a potent downregulation of migratory and invasive capacity independent of their BRAF/NRAS mutational status. Furthermore, the combination attenuated tumor growth rate, as ascertained by the significant reduction of Ki67 expression and induced DNA damage in vivo. Importantly, this combination therapy had minimal therapy-related toxicity in mice. Lastly, the cell cycle G2 checkpoint kinase WEE1 and the RTK IGF1R, non-canonical targets, were altered upon exposure to the combination. Knockdown of WEE1 abrogated the combination-mediated effects on cell migration and proliferation in BRAF mutant BRAF inhibitor-sensitive cells, whereas WEE1 silencing alone inhibited cell migration in NRAS mutant cells. In summary, our results show that afatinib and crizotinib in combination is a promising alternative targeted therapy option for CMM patients, irrespective of BRAF/NRAS mutational status, as well as for cases where resistance has developed towards BRAF inhibitors.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Melanoma/tratamento farmacológico , Proteínas Proto-Oncogênicas c-met/antagonistas & inibidores , Neoplasias Cutâneas/tratamento farmacológico , Afatinib/farmacologia , Animais , Linhagem Celular Tumoral , Crizotinibe/farmacologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Receptores ErbB/metabolismo , Feminino , GTP Fosfo-Hidrolases/genética , GTP Fosfo-Hidrolases/metabolismo , Humanos , Melanoma/genética , Melanoma/metabolismo , Melanoma/patologia , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos SCID , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Proteínas Proto-Oncogênicas B-raf/metabolismo , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
There are few reports of breast cancer cases with uterine metastases. Here, we report a metastatic lobular carcinoma to endometrium presenting as abnormal uterine bleeding. Diagnosis was based in previous lobular breast carcinoma and immunohistochemistry.
Assuntos
Coinfecção , Histoplasmose , Humanos , Antifúngicos/uso terapêutico , Coinfecção/microbiologia , Dermatomicoses/tratamento farmacológico , Dermatomicoses/microbiologia , Histoplasma/isolamento & purificação , Histoplasmose/tratamento farmacológico , Histoplasmose/diagnóstico , Combinação Trimetoprima e Sulfametoxazol/uso terapêuticoRESUMO
We present a new facet of isobenzofuran chemistry which allows for its efficient manipulation to generate biologically relevant entities. This methodology has been successfully applied toward the synthesis of ajudazol A.