RESUMO
In the title compound, C14H11ClN2O2S, the dihedral angle between the pyrrolo-[1,2-c]pyrimidine ring system (r.m.s. deviation = 0.008â Å) and the benzene ring is 80.2â (9)°. In the crystal, inversion dimers linked by pairs of C-Hâ¯O inter-actions generate R 2 2(16) loops. Several aromatic π-π stacking inter-actions between the pyrrolo-[1,2-c]pyrimidine rings, as well as separately between the pyrrolo and pyrimidine groups [shortest centroid-centroid separation = 3.5758â (14)â Å], help to consolidate the packing.
RESUMO
Two new analogues of CDDO-Imidazolide (CDDO-Im), namely 1-[2-Cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]-4(-pyridin-2-yl)-1H-imidazole ("CDDO-2P-Im") and 1-[2-Cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]-4(-pyridin-3-yl)-1H-imidazole ("CDDO-3P-Im") have been synthesized and tested for their potential use as chemopreventive drugs. At nanomolar concentrations, they were equipotent to CDDO-Im for inducing differentiation and apoptosis in U937 leukemia cells. As inflammation and oxidative stress contribute to carcinogenesis, we also assessed their cytoprotective potential. The new compounds suppressed inducible nitric oxide synthase (iNOS) expression in RAW264.7 macrophage-like cells and significantly elevated heme oxygenase-1 (HO-1) and quinone reductase (NQO1) mRNA and protein levels in various mouse tissues in vivo. Most importantly, pharmacokinetic studies performed in vitro in human plasma and in vivo showed that each new analogue was more stable than CDDO-Im. Much higher concentrations of the new derivatives were found in mouse liver, lung, pancreas and kidney after gavage in contrast to CDDO-Im. Because of their better bioavailability and their excellent anti-inflammatory profile in vitro, CDDO-2P-Im and CDDO-3P-Im were tested for prevention in a highly relevant mouse lung cancer model, in which A/J mice develop lung carcinomas after injection of vinyl carbamate, a potent carcinogen. CDDO-2P-Im and CDDO-3P-Im were as effective as CDDO-Im for reducing the size and the severity of the lung tumors.
Assuntos
Anticarcinógenos/farmacologia , Antineoplásicos/farmacologia , Imidazóis/farmacologia , Neoplasias/prevenção & controle , Ácido Oleanólico/análogos & derivados , Animais , Linhagem Celular Tumoral , Células Cultivadas , Feminino , Heme Oxigenase-1/metabolismo , Humanos , Camundongos , NAD(P)H Desidrogenase (Quinona)/metabolismo , Neoplasias/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ácido Oleanólico/farmacologia , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/metabolismo , Células U937RESUMO
This review presents the biological activity-antibacterial, antifungal, anti-parasitic, antiviral, antitumor, antiinflammatory, antioxidant, and enzymatic activity-of halogenated marine natural products discovered in the past five years. Newly discovered examples that do not report biological activity are not included.
Assuntos
Organismos Aquáticos/química , Produtos Biológicos/farmacologia , Descoberta de Drogas , Hidrocarbonetos Halogenados/farmacologia , Animais , Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Antifúngicos/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Antiparasitários/farmacologia , Antivirais/farmacologia , HumanosRESUMO
An efficient synthesis of methyl 2-cyano-3,12-dioxoursol-1,9-dien-28-oate (CDDU-methyl ester) from commercially available ursolic acid, which features an oxidative ozonolysis-mediated C-ring enone formation, and provides the first access to ursolic acid-derived cyano enone analogues with C-ring activation. These new ursolic acid analogues show potent biological activities, with potency of approximately five-fold less than the corresponding oleanolic acid derivatives.
Assuntos
Anti-Infecciosos/síntese química , Ácido Oleanólico/química , Triterpenos Pentacíclicos/síntese química , Triterpenos/química , Animais , Anti-Infecciosos/farmacologia , Linhagem Celular , Ésteres , Concentração Inibidora 50 , Interferon gama/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Camundongos , Estrutura Molecular , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Ácido Oleanólico/farmacologia , Triterpenos Pentacíclicos/farmacologia , Triterpenos/farmacologia , Ácido UrsólicoRESUMO
2-Cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO, 2) was condensed with various amino acid methyl esters at the C-28 carboxylic acid. The new amide conjugates were evaluated for their inhibition of nitric oxide (NO) production in RAW264.7 cells stimulated with interferon-γ (IFNγ). Of these new compounds, CDDO conjugates with alanine, valine, and serine are nearly equipotent to CDDO-ethyl amide (4), a triterpenoid with promising biological activity in numerous disease models. Some of these conjugates also induce the in vitro expression of heme oxygenase-1, and inhibit the proliferation of Panc-1343 pancreatic cells.
Assuntos
Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Ácido Oleanólico/análogos & derivados , Aminoácidos/química , Aminoácidos/farmacologia , Animais , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Camundongos , Ácido Oleanólico/química , Ácido Oleanólico/farmacologiaRESUMO
Bardoxolone methyl (2-cyano-3,12-dioxooleane-1,9(11)-dien-28-oic acid methyl ester; CDDO-Me) (1), a synthetic oleanane triterpenoid with highly potent anti-inflammatory activity (levels below 1 nM), has completed a successful phase I clinical trial for the treatment of cancer and a successful phase II trial for the treatment of chronic kidney disease in type 2 diabetes patients. Our synthesis of bardoxolone methyl (1) proceeds in â¼50% overall yield in five steps from oleanolic acid (2), requires only one to two chromatographic purifications, and can provide gram quantities of 1.
Assuntos
Anti-Inflamatórios/síntese química , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/síntese química , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Cromatografia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Ésteres , Humanos , Estrutura Molecular , Ácido Oleanólico/química , Ácido Oleanólico/farmacologia , Insuficiência Renal Crônica/tratamento farmacológicoRESUMO
Forty-four novel tricycles containing nonenolizable cyano enones (TCEs) were designed and synthesized on the basis of a semisynthetic pentacyclic triterpenoid, bardoxolone methyl, which is currently being developed in phase II clinical trials for the treatment of severe chronic kidney disease in diabetic patients. Most of the TCEs having two different kinds of nonenolizable cyano enones in rings A and C are highly potent suppressors of induction of inducible nitric oxide synthase stimulated with interferon-γ and are highly potent inducers of the cytoprotective enzymes heme oxygenase-1 and NAD(P)H:quinone oxidoreductase-1. Among these compounds, (±)-(4bS,8aR,10aS)-10a-ethynyl-4b,8,8-trimethyl-3,7-dioxo-3,4b,7,8,8a,9,10,10a-octahydrophenanthrene-2,6-dicarbonitrile ((±)-31) is the most potent in these bioassays in our pool of drug candidates including semisynthetic triterpenoids and synthetic tricycles. These facts strongly suggest that an essential factor for potency is not a triterpenoid skeleton but the cyano enone functionality. Notably, TCE 31 reduces hepatic tumorigenesis induced with aflatoxin in rats. Further preclinical studies and detailed mechanism studies on 31 are in progress.
Assuntos
Anti-Inflamatórios não Esteroides/síntese química , Antineoplásicos/síntese química , Nitrilas/síntese química , Fenantrenos/síntese química , Aflatoxina B1 , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Células Cultivadas , Citoproteção , Indução Enzimática , Heme Oxigenase-1/biossíntese , Interferon gama/farmacologia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Neoplasias Hepáticas Experimentais/induzido quimicamente , Neoplasias Hepáticas Experimentais/prevenção & controle , Macrófagos/efeitos dos fármacos , Macrófagos/enzimologia , Masculino , Camundongos , NAD(P)H Desidrogenase (Quinona)/biossíntese , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/biossíntese , Nitrilas/química , Nitrilas/farmacologia , Triterpenos Pentacíclicos/química , Fenantrenos/química , Fenantrenos/farmacologia , Lesões Pré-Cancerosas/induzido quimicamente , Lesões Pré-Cancerosas/prevenção & controle , Ratos , Estereoisomerismo , Estômago/efeitos dos fármacos , Estômago/enzimologia , Relação Estrutura-AtividadeRESUMO
A novel acetylenic tricyclic bis-(cyano enone), TBE-31, is a lead compound in a series of tricyclic compounds with enone functionalities in rings A and C. Nanomolar concentrations of this potent multifunctional molecule suppress the induction of the inflammatory protein, inducible nitric oxide synthase, activate phase 2 cytoprotective enzymes in vitro and in vivo, block cell proliferation, and induce differentiation and apoptosis of leukemia cells. Oral administration of TBE-31 also significantly reduces formation of aflatoxin-DNA adducts and decreases size and number of aflatoxin-induced preneoplastic hepatic lesions in rats by >90%. Because of the two cyano enones in rings A and C, TBE-31 may directly interact with DTT and protein targets such as Keap1 that contain reactive cysteine residues. The above findings suggest that TBE-31 should also be tested for chemoprevention and chemotherapy in relevant models of cancer and against other chronic, degenerative diseases in which inflammation and oxidative stress contribute to disease pathogenesis.
Assuntos
Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Transformação Celular Neoplásica/efeitos dos fármacos , Neoplasias Hepáticas/prevenção & controle , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Fenantrenos/farmacologia , Administração Oral , Aflatoxina B1/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Adutos de DNA/metabolismo , Heme Oxigenase-1/metabolismo , Humanos , Imidazóis/farmacologia , Leucemia/tratamento farmacológico , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/patologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Estrutura Molecular , NAD(P)H Desidrogenase (Quinona)/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/farmacologia , Fenantrenos/química , Ratos , Ratos Endogâmicos F344 , Espécies Reativas de Oxigênio/metabolismoRESUMO
PURPOSE: To test whether the triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me) and the rexinoid LG100268 (268) prevent the formation of estrogen receptor (ER)-negative mammary tumors or either arrest the growth or cause regression of established tumors in MMTV-neu mice. EXPERIMENTAL DESIGN: For prevention, mice were fed control diet, CDDO-Me (60 mg/kg diet), 268 (20 mg/kg diet), or the combination for 45 weeks. For treatment, mice with established tumors at least 4 mm in diameter were fed control diet, CDDO-Me (100 mg/kg diet), 268 (60 mg/kg diet), or the combination for 4 weeks. RESULTS: CDDO-Me and 268 significantly delayed the development of ER-negative tumors, with a 14- and 24-week delay, respectively, compared with the control group for the time required to reach 50% tumor incidence. The combination of CDDO-Me and 268 was significantly more potent than the individual drugs, as only one tumor was found in the combination group, after 45 weeks on diet, at which time all control animals had tumors. Treating established tumors with CDDO-Me arrested the growth of 86% of the tumors, and 268 induced tumor regression in 85% of tumors. CDDO-Me and 268 target different signaling pathways and cell types. CDDO-Me inhibited constitutive STAT3 phosphorylation and the degradation of IKBalpha in ER-negative breast cancer cells, whereas 268 blocked IKBalpha degradation and the release of interleukin-6 in RAW264.7 macrophage-like cells, inhibited the ability of endothelial cells to organize into networks, and blocked angiogenesis in vivo. CONCLUSIONS: CDDO-Me and 268 are useful as individual drugs to prevent ER-negative mammary tumorigenesis and to treat established tumors. They synergize when used in combination for prevention.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Neoplasias Mamárias Experimentais/tratamento farmacológico , Ácidos Nicotínicos/farmacologia , Ácido Oleanólico/análogos & derivados , Tetra-Hidronaftalenos/farmacologia , Administração Oral , Animais , Antineoplásicos/farmacologia , Movimento Celular/efeitos dos fármacos , Sinergismo Farmacológico , Células Endoteliais/efeitos dos fármacos , Feminino , Humanos , Vírus do Tumor Mamário do Camundongo/genética , Camundongos , Camundongos Transgênicos , Neovascularização Patológica/tratamento farmacológico , Ácido Oleanólico/farmacologia , Receptor ErbB-2/genética , Receptores de Estrogênio/biossínteseRESUMO
Female A/J mice injected with the carcinogen vinyl carbamate develop atypical adenomatous hyperplasias in lungs 4 weeks after injection with the carcinogen. The number and severity of tumors then increase over time, making these mice a useful model for evaluating potential chemopreventive agents. The rexinoid LG100268 (LG268), a selective ligand for the retinoid X receptor, and the methyl amide of the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO) both significantly reduced the number, size, and severity of the histopathology of lung tumors in female A/J mice when fed in diet for 14 to 20 weeks. The total tumor burden was 85% to 87% lower in mice fed LG268 and CDDO-MA than in controls, and the percentage of high-grade tumors decreased from 59% in the controls to 25% or 30% with CDDO-MA and LG268. Erlotinib, which is used to treat lung cancer patients and is an inhibitor of the epidermal growth factor receptor, was less effective in this model. Immunohistochemical staining of geminin, a marker of cell cycle progression, was higher in lung sections from control mice than in mice treated with LG268. Because rexinoids and triterpenoids signal through different biological pathways, they should be tested in combination for the prevention of lung cancer.
Assuntos
Anticarcinógenos/uso terapêutico , Neoplasias Pulmonares/prevenção & controle , Pulmão/efeitos dos fármacos , Ácidos Nicotínicos/uso terapêutico , Ácido Oleanólico/análogos & derivados , Quinazolinas/uso terapêutico , Tetra-Hidronaftalenos/uso terapêutico , Animais , Linhagem Celular Tumoral , Cloridrato de Erlotinib , Feminino , Neoplasias Pulmonares/patologia , Camundongos , Ácido Oleanólico/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêuticoRESUMO
The known cembranoid diterpene eunicin (1) was isolated from a new genus of octocoral, Pseudoplexaura, collected in the Caribbean. The complete assignment of the 13C and 1H NMR spectra, including differentiation of pro-R and pro-S hydrogens and elucidation of all J values, was accomplished with the aid of TOCSY, HSQC, and HMBC data, and a consideration of three low-energy conformations using SYBYL.
Assuntos
Antozoários/química , Antifúngicos/química , Diterpenos/química , Animais , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Linhagem Celular Tumoral , Diterpenos/isolamento & purificação , Diterpenos/farmacologia , Humanos , Imunodifusão , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Camundongos , Modelos Moleculares , Conformação Molecular , Rotação Ocular , Staphylococcus aureus/efeitos dos fármacosRESUMO
The anti-tumor synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO)-imidazolide (CDDO-Im) ectopically activates the transforming growth factor beta (TGFbeta)-Smad pathway and extends the duration of signaling by an undefined mechanism. Here we show that CDDO-Imdependent persistence of Smad2 phosphorylation is independent of Smad2 phosphatase activity and correlates with delayed TGFbeta receptor degradation and trafficking. Altered TGFbeta trafficking parallels the dispersal of EEA1-positive endosomes from the perinuclear region of CDDO-Im-treated cells. The effect of CDDO-Im on the EEA1 compartment led to an analysis of the cytoskeleton, and we observed that CDDO-Im alters microtubule dynamics by disrupting the microtubule-capping protein, Clip-170. Interestingly, biotinylated triterpenoid was found to localize to the polarity complex at the leading edge of migrating cells. Furthermore, CDDO-Im disrupted the localization of IQGAP1, PKCzeta, Par6, and TGFbeta receptors from the leading edge of migrating cells and inhibited TGFbeta-dependent cell migration. Thus, the synthetic triterpenoid CDDO-Im interferes with TGFbeta receptor trafficking and turnover and disrupts cell migration by severing the link between members of the polarity complex and the microtubule network.
Assuntos
Citoesqueleto/metabolismo , Imidazóis/farmacologia , Ácido Oleanólico/análogos & derivados , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Transporte Ativo do Núcleo Celular , Animais , Células COS , Movimento Celular , Núcleo Celular/metabolismo , Chlorocebus aethiops , Fibroblastos/metabolismo , Microtúbulos/metabolismo , Modelos Biológicos , Ácido Oleanólico/farmacologia , RatosRESUMO
Inducible phase 2 enzymes constitute a primary line of cellular defense. The oleanane dicyanotriterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-onitrile (TP-225) is a very potent inducer of these systems. Topical application of TP-225 to SKH-1 hairless mice increases the levels of NAD(P)H-quinone acceptor oxidoreductase 1 (NQO1) and heme oxygenase 1 (HO-1) and protects against UV radiation-induced dermal thickening. Daily topical treatments of 10 nmol of TP-225 to the backs of mice that were previously subjected to low-level chronic UVB radiation (30 mJ/cm(2)/session, twice a week for 17 weeks), led to 50% reduction in multiplicity of skin tumors. In addition, the total tumor burden of squamous cell carcinomas was reduced by 5.5-fold. The identification of new agents for protection against UV radiation-induced skin cancer and understanding of their mechanism(s) of action is especially important in view of the fact that human skin cancers represent a significant source of increasing morbidity and mortality.
Assuntos
Heme Oxigenase-1/metabolismo , NADPH Desidrogenase/metabolismo , Neoplasias Induzidas por Radiação/tratamento farmacológico , Neoplasias Induzidas por Radiação/metabolismo , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/metabolismo , Triterpenos/administração & dosagem , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Citoproteção/efeitos dos fármacos , Feminino , Camundongos , Camundongos Pelados , NAD(P)H Desidrogenase (Quinona) , Neoplasias Induzidas por Radiação/etiologia , Neoplasias Induzidas por Radiação/patologia , Neoplasias Cutâneas/etiologia , Neoplasias Cutâneas/patologia , Raios Ultravioleta/efeitos adversosRESUMO
Betulinic acid (BA), a pentacyclic triterpene isolated from birch bark and other plants, selectively inhibits the growth of human cancer cell lines. However, the poor potency of BA hinders its clinical development, despite a lack of toxicity in animal studies even at high concentrations. Here, we describe six BA derivatives that are markedly more potent than BA for inhibiting inducible nitric oxide synthase, activating phase 2 cytoprotective enzymes, and inducing apoptosis in cancer cells and in Bax/Bak(-/-) fibroblasts, which lack two key proteins involved in the intrinsic, mitochondrial-dependent apoptotic pathway. Notably, adding a cyano-enone functionality in the A ring of BA enhanced its cytoprotective properties, but replacing the cyano group with a methoxycarbonyl strikingly increased potency in the apoptosis assays. Higher plasma and tissue levels were obtained with the new BA analogues, especially CBA-Im [1-(2-cyano-3-oxolupa-1,20(29)-dien-28-oyl)imidazole], compared with BA itself and at concentrations that were active in vitro. These results suggest that BA is a useful platform for drug development, and the enhanced potency and varied biological activities of CBA-Im make it a promising candidate for further chemoprevention or chemotherapeutic studies.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Citoproteção/efeitos dos fármacos , Triterpenos/farmacologia , Animais , Antineoplásicos Fitogênicos/sangue , Antineoplásicos Fitogênicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Inflamação , Macrófagos/efeitos dos fármacos , Macrófagos/enzimologia , Masculino , Camundongos , NAD(P)H Desidrogenase (Quinona)/metabolismo , Óxido Nítrico/biossíntese , Triterpenos Pentacíclicos , Triterpenos/sangue , Triterpenos/química , Proteína X Associada a bcl-2/metabolismo , Ácido BetulínicoRESUMO
We report the first use of new synthetic triterpenoids to prevent lung cancer in experimental animals. Female A/J mice were treated with the mutagenic carcinogen vinyl carbamate, which induces adenocarcinoma of the lung in all animals within 16 weeks. If mice were fed either the methyl ester or the ethyl amide derivative of the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO-ME and CDDO-EA, respectively), beginning 1 week after dosing with carcinogen, the number, size, and severity of lung carcinomas were markedly reduced. The mechanisms of action of CDDO-ME and CDDO-EA that are germane to these in vivo findings are the following results shown here in cell culture: (a) suppression of the ability of IFN-gamma to induce de novo formation of nitric oxide synthase in a macrophage-like cell line RAW264.7, (b) induction of heme oxygenase-1 in these RAW cells, and (c) suppression of phosphorylation of the transcription factor signal transducers and activators of transcription 3 as well as induction of apoptosis in human lung cancer cell lines.
Assuntos
Adenocarcinoma/prevenção & controle , Anticarcinógenos/farmacologia , Neoplasias Pulmonares/prevenção & controle , Ácido Oleanólico/análogos & derivados , Adenocarcinoma/induzido quimicamente , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Animais , Anticarcinógenos/sangue , Anticarcinógenos/farmacocinética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Pulmão/metabolismo , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Camundongos , Camundongos Endogâmicos A , Ácido Oleanólico/sangue , Ácido Oleanólico/farmacocinética , Ácido Oleanólico/farmacologia , Fosforilação , Fator de Transcrição STAT3/metabolismo , Uretana/análogos & derivadosRESUMO
Novel C-8a functionalized tricyclic compounds having cyano enones in rings A and C have been synthesized and biologically evaluated. Among them, compounds with acetylene groups at C-8a show extremely high potency in in vitro and in vivo bioassays for anti-inflammatory and cytoprotective activities. Both in vitro and in vivo potencies are markedly higher than those of 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO), which is being evaluated as an anticancer drug in phase I clinical trials.
Assuntos
Alcinos/síntese química , Anti-Inflamatórios não Esteroides/síntese química , Antineoplásicos/síntese química , Compostos Heterocíclicos com 3 Anéis/síntese química , Nitrilas/síntese química , Ácido Oleanólico/análogos & derivados , Alcinos/farmacologia , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/farmacologia , Linhagem Celular , Citoproteção , Compostos Heterocíclicos com 3 Anéis/farmacologia , Interferon gama/farmacologia , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Nitrilas/farmacologia , Ácido Oleanólico/síntese química , Ácido Oleanólico/farmacologia , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: Receptor activator of nuclear factor-kappaB ligand (RANKL) promotes osteoclast differentiation from monocyte precursors by inducing a cohort of genes, including tartrate-resistant acid phosphatase (TRAP) and matrix metalloproteinase-9 (MMP-9). A family of synthetic triterpenoids with antiinflammatory and pro-apoptotic properties was described to modulate differentiation in monocytic cell lineages. We therefore investigated the ability of the potent and bioavailable synthetic triterpenoid TP-222 to inhibit RANKL-induced osteoclast formation and MMP-9 expression from monocytic precursor cells. METHODS: Osteoclast formation was assayed by staining for TRAP-positive multinucleated cells. MMP-9 expression was measured by quantitative RT-PCR, Western blot, immunohistochemistry, and gel zymography. In vivo effects of TP-222 were assessed by daily intraperitoneal injection of 4-week-old mice for 7 days followed by measurement of osteoclast number and MMP-9 expression at the cartilage/bone junction of the epiphyseal growth plate. RESULTS: RANKL promoted and TP-222 (300 nM) inhibited osteoclast formation in cultures of RAW264.7 cells or bone marrow-derived monocytes. RANKL also induced MMP-9 expression in RAW264.7 cells and this was reduced by concurrent or subsequent addition of TP-222. TP-222 treatment significantly reduced the mean number of osteoclasts present at the cartilage/bone interface compared to vehicle-injected control mice. Morphometric analyses of tissue sections showed that TP-222 treatment reduced the amount of immunoreactive MMP-9 present in both mononucleated pre-osteoclasts and osteoclasts. CONCLUSION: Our data demonstrate that TP-222 inhibits osteoclast formation and MMP-9 expression in vitro and in vivo, and suggest that triterpenoids may be useful compounds for modulating bone resorption diseases.
Assuntos
Metaloproteinase 9 da Matriz/metabolismo , Osteoclastos/efeitos dos fármacos , Ligante RANK/antagonistas & inibidores , Triterpenos/farmacologia , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/enzimologia , Contagem de Células , Linhagem Celular , Combinação de Medicamentos , Inibidores Enzimáticos/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Injeções Intraperitoneais , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Metaloproteinase 9 da Matriz/genética , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Óxido Nítrico/antagonistas & inibidores , Osteoclastos/enzimologia , Osteogênese/efeitos dos fármacos , RNA Mensageiro/metabolismoRESUMO
Synthetic triterpenoids have been developed, which are potent inducers of cytoprotective enzymes and inhibitors of inflammation, greatly improving on the weak activity of naturally occurring triterpenoids. An imidazolide triterpenoid derivative, 1-[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Im or TP235), has been previously shown to potently protect against hepatic tumorigenesis, acting in part by inducing cytoprotective genes through Keap1-Nrf2-antioxidant response element (ARE) signaling. In these studies, the pharmacodynamic activity of CDDO-Im is characterized in two distinct lines of ARE reporter mice and by measuring increases in Nqo1 transcript levels as a marker of cytoprotective gene induction. Oral administration of CDDO-Im induces ARE-regulated cytoprotective genes in many tissues in the mouse, including liver, lung, kidney, intestines, brain, heart, thymus, and salivary gland. CDDO-Im induces Nqo1 RNA transcripts in some organs at doses as low as 0.3 mumol/kg body weight (orally). A structure activity evaluation of 15 additional triterpenoids (a) confirmed the importance of Michael acceptor groups on both the A and C rings, (b) showed the requirement for a nitrile group at C-2 of the A ring, and (c) indicated that substituents at C-17 dramatically affected pharmacodynamic action in vivo. In addition to CDDO-Im, other triterpenoids, particularly the methyl ester CDDO-Me (TP155) and the dinitrile TP225, are extremely potent inducers of cytoprotective genes in mouse liver, lung, small intestine mucosa, and cerebral cortex. This pharmacodynamic characterization highlights the chemopreventive promise of several synthetic triterpenoids in multiple target organs.
Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/genética , Triterpenos/farmacologia , Administração Oral , Animais , Antioxidantes/metabolismo , Quimioprevenção , Relação Dose-Resposta a Droga , Genes Reporter/genética , Imidazóis/química , Fígado/citologia , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/patologia , Masculino , Camundongos , NAD(P)H Desidrogenase (Quinona) , NADPH Desidrogenase/genética , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/química , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Elementos de Resposta/genética , Ativação Transcricional , Triterpenos/administração & dosagem , Triterpenos/químicaRESUMO
The synthetic triterpenoid 1-[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Im) is a multifunctional agent with potent anti-inflammatory, antiproliferative, cytoprotective, and apoptotic activities, whose molecular targets are unknown. Using both cell-free and cellular assays, we show that CDDO-Im is a direct inhibitor of IkappaB kinase (IKK) beta and that it thereby inhibits binding of nuclear factor-kappaB to DNA and subsequent transcriptional activation. Pretreatment of cells with CDDO-Im prevents IkappaBalpha phosphorylation and degradation in response to tumor necrosis factor alpha. The kinetics of this inhibition by CDDO-Im are rapid and occur within 15 min. A biotinylated analogue of CDDO-Im showed that CDDO-Im binds to the IKK signalsome. Furthermore, we show that Cys(179) on IKK is a target for CDDO-Im. This is the first report to show that this novel synthetic triterpenoid binds to and inhibits IKKbeta directly.
Assuntos
Quinase I-kappa B/antagonistas & inibidores , Imidazóis/farmacologia , NF-kappa B/metabolismo , Ácido Oleanólico/análogos & derivados , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Cisteína/química , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , Ácido Oleanólico/farmacologia , Compostos de Sulfidrila/químicaRESUMO
Fifteen new betulinic acid analogues were designed, synthesized, and tested for anti-inflammatory activity. Many of these analogues effectively suppress nitric oxide (NO) production in RAW cells stimulated with interferon-gamma. Analogue 10 is highly and orally active in vivo for induction of the anti-inflammatory and cytoprotective enzyme, heme oxygenase-1.