RESUMO
DNA is a key target for anticancer and antimicrobial drugs. Assessing the bioactivity of compounds involves in silico and instrumental studies to determine their affinity for biomolecules like DNA. This study explores the potential of the switchSense technique in rapidly evaluating compound bioactivity towards DNA. By combining switchSense with computational methods and UV-Vis spectrophotometry, various bioactive compounds' interactions with DNA were analyzed. The objects of the study were: netropsin (as a model compound that binds in the helical groove), as well as derivatives of pyrazine (PTCA), sulfonamide (NbutylS), and anthraquinone (AQ-NetOH). Though no direct correlation was found between switchSense kinetics and binding modes, this research suggests the technique's broader utility in assessing new compounds' interactions with DNA. used as analytes whose interactions with DNA have not been yet fully described in the literature.
Assuntos
Antraquinonas , DNA , Espectrofotometria Ultravioleta , DNA/química , DNA/metabolismo , Antraquinonas/química , Antraquinonas/farmacologia , Netropsina/química , Netropsina/metabolismo , Netropsina/farmacologia , Sulfonamidas/química , Sulfonamidas/farmacologia , Sulfonamidas/metabolismo , Cinética , Simulação de Acoplamento MolecularRESUMO
A stable intense resistance called "nonhost resistance" generates a complete multiple-gene resistance against plant pathogenic species that are not pathogens of pea such as the bean pathogen, Fusarium solani f. sp. phaseoli (Fsph). Chitosan is a natural nonhost resistance response gene activator of defense responses in peas. Chitosan may share with cancer-treatment compounds, netropsin and some anti-cancer drugs, a DNA minor groove target in plant host tissue. The chitosan heptamer and netropsin have the appropriate size and charge to reside in the DNA minor groove. The localization of a percentage of administered radio-labeled chitosan in the nucleus of plant tissue in vivo indicates its potential to transport to site(s) within the nuclear chromatin (1,2). Other minor groove-localizing compounds administered to pea tissue activate the same secondary plant pathway that terminates in the production of the anti-fungal isoflavonoid, pisatin an indicator of the generated resistance response. Some DNA minor groove compounds also induce defense genes designated as "pathogenesis-related" (PR) genes. Hypothetically, DNA targeting components alter host DNA in a manner enabling the transcription of defense genes previously silenced or minimally expressed. Defense-response-elicitors can directly (a) target host DNA at the site of transcription or (b) act by a series of cascading events beginning at the cell membrane and indirectly influence transcription. A single defense response, pisatin induction, induced by chitosan and compounds with known DNA minor groove attachment potential was followed herein. A hypothesis is formulated suggesting that this DNA target may be accountable for a portion of the defense response generated in nonhost resistance.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Quitosana/farmacologia , Substâncias Intercalantes/farmacologia , Netropsina/farmacologia , Pisum sativum/genética , Doenças das Plantas/genética , Pterocarpanos/farmacologia , Antineoplásicos Fitogênicos/química , Benzimidazóis/química , Benzimidazóis/farmacologia , Quitosana/química , Cromatina/química , Cromatina/efeitos dos fármacos , Cromatina/metabolismo , Cromomicinas/química , Cromomicinas/farmacologia , DNA de Plantas/genética , DNA de Plantas/metabolismo , Resistência à Doença/genética , Fusarium/crescimento & desenvolvimento , Fusarium/patogenicidade , Regulação da Expressão Gênica de Plantas , Proteínas HMGA/genética , Proteínas HMGA/metabolismo , Substâncias Intercalantes/química , Netropsina/química , Pisum sativum/imunologia , Pisum sativum/metabolismo , Pisum sativum/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Pterocarpanos/química , Transcrição GênicaRESUMO
The DNA as the depository of genetic information is a natural target for chemotherapy. A lot of anticancer and antimicrobial agents derive their biological activity from their selective interaction with DNA in the minor groove and from their ability to interfere with biological processes such as enzyme catalysis, replication and transcription. The discovery of the details of minor groove binding drugs, such as netropsin and distamycin A, oligoamides built of 4-amino-1-methylpyrrole-2-carboxylic acid residues, allowed to develop various DNA sequence-reading molecules, named lexitropsins, capable of interacting with DNA precisely, strongly and with a high specificity, and at the same time exhibiting significant cytotoxic potential. Among such compounds, lexitropsins built of carbocyclic sixmembered aromatic rings occupy a quite prominent place in drug research. This work is an attempt to present current findings in the study of carbocyclic lexitropins, their structures, syntheses and biological investigations such as DNA-binding and antiproliferative activity.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Distamicinas/química , Distamicinas/farmacologia , Desenho de Fármacos , Netropsina/análogos & derivados , Netropsina/farmacologia , Ácidos Carbocíclicos/síntese química , Ácidos Carbocíclicos/química , Ácidos Carbocíclicos/farmacologia , Animais , Antibacterianos/síntese química , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , DNA/química , DNA/metabolismo , Distamicinas/síntese química , Humanos , Neoplasias/tratamento farmacológico , Netropsina/síntese químicaRESUMO
Malaria caused by Plasmodium falciparum is the major disease burden all over the world. Recently, the situation has deteriorated because the malarial parasites are becoming progressively more resistant to numerous commonly used antimalarial drugs. Thus, there is a critical requirement to find other means to restrict and eliminate malaria. The mismatch repair (MMR) machinery of parasite is quite unique in several ways, and it can be exploited for finding new drug targets. MutL homolog (MLH) is one of the major components of MMR machinery, and along with UvrD, it helps in unwinding the DNA. We have screened several DNA-interacting ligands for their effect on intrinsic ATPase activity of PfMLH protein. This screening suggested that several ligands such as daunorubicin, etoposide, ethidium bromide, netropsin, and nogalamycin are inhibitors of the ATPase activity of PfMLH, and their apparent IC50 values range from 2.1 to 9.35 µM. In the presence of nogalamycin and netropsin, the effect was significant because in their presence, the V max value dropped from 1.024 µM of hydrolyzed ATP/min to 0.596 and 0.643 µM of hydrolyzed ATP/min, respectively. The effect of double-stranded RNAs of PfMLH and PfUvrD on growth of P. falciparum 3D7 strain was studied. The parasite growth was significantly inhibited suggesting that these components belonging to MMR pathway are crucial for the survival of the parasite.
Assuntos
Adenosina Trifosfatases/antagonistas & inibidores , Antimaláricos/farmacologia , DNA Helicases/metabolismo , Reparo de Erro de Pareamento de DNA/efeitos dos fármacos , Malária Falciparum/tratamento farmacológico , Proteína 1 Homóloga a MutL/metabolismo , Plasmodium falciparum/crescimento & desenvolvimento , RNA de Cadeia Dupla/farmacologia , Adenosina Trifosfatases/metabolismo , Reparo de Erro de Pareamento de DNA/genética , DNA de Protozoário/genética , Daunorrubicina/farmacologia , Resistência a Medicamentos , Etídio/farmacologia , Etoposídeo/farmacologia , Malária Falciparum/parasitologia , Simulação de Acoplamento Molecular , Proteína 1 Homóloga a MutL/genética , Netropsina/farmacologia , Nogalamicina/farmacologia , Plasmodium falciparum/genética , Plasmodium falciparum/metabolismoRESUMO
HMGA2 is an important chromatin factor that interacts with DNA via three AT-hook domains, thereby regulating chromatin architecture and transcription during embryonic and fetal development. The protein is absent from differentiated somatic cells, but aberrantly re-expressed in most aggressive human neoplasias where it is causally linked to cell transformation and metastasis. DNA-binding also enables HMGA2 to protect cancer cells from DNA-damaging agents. HMGA2 therefore is considered to be a prime drug target for many aggressive malignancies. Here, we have developed a broadly applicable cell-based reporter system which can identify HMGA2 antagonists targeting functionally important protein domains, as validated with the known AT-hook competitor netropsin. In addition, high-throughput screening can uncover functional links between HMGA2 and cellular factors important for cell transformation. This is demonstrated with the discovery that HMGA2 potentiates the clinically important topoisomerase I inhibitor irinotecan/SN-38 in trapping the enzyme in covalent DNA-complexes, thereby attenuating transcription.
Assuntos
Camptotecina/análogos & derivados , DNA Topoisomerases Tipo I/fisiologia , Proteína HMGA2/fisiologia , Inibidores da Topoisomerase I/farmacologia , Sequência de Aminoácidos , Camptotecina/farmacologia , Diferenciação Celular/efeitos dos fármacos , Estabilidade Enzimática , Genes Reporter , Células HEK293 , Células HeLa , Ensaios de Triagem em Larga Escala , Humanos , Irinotecano , Luciferases de Renilla/biossíntese , Luciferases de Renilla/genética , Netropsina/farmacologia , Regiões Promotoras Genéticas , Transcrição Gênica/efeitos dos fármacos , Ativação TranscricionalRESUMO
Acute lymphoblastic leukaemia (ALL) in infants is an intractable cancer in childhood. Although recent intensive chemotherapy progress has considerably improved ALL treatment outcome, disease cure is often accompanied by undesirable long-term side effects, and efficient, less toxic molecular targeting therapies have been anticipated. In infant ALL cells with KMT2A (MLL) fusion, the microRNA let-7b (MIRLET7B) is significantly downregulated by DNA hypermethylation of its promoter region. We show here that the expression of HMGA2, one of the oncogenes repressed by MIRLET7B, is reversely upregulated in infant ALL leukaemic cells, particularly in KMT2A-AFF1 (MLL-AF4) positive ALL. In addition to the suppression of MIRLET7B, KMT2A fusion proteins positively regulate the expression of HMGA2. HMGA2 is one of the negative regulators of CDKN2A gene, which encodes the cyclin-dependent kinase inhibitor p16(INK4A) . The HMGA2 inhibitor netropsin, when combined with demethylating agent 5-azacytidine, upregulated and sustained the expression of CDKN2A, which resulted in growth suppression of KMT2A-AFF1-expressing cell lines. This effect was more apparent compared to treatment with 5-azacytidine alone. These results indicate that the MIRLET7B-HMGA2-CDKN2A axis plays an important role in cell proliferation of leukaemic cells and could be a possible molecular target for the therapy of infant ALL with KMT2A-AFF1.
Assuntos
Proteína HMGA2/antagonistas & inibidores , MicroRNAs/farmacologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Azacitidina/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/fisiologia , Metilação de DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Sinergismo Farmacológico , Técnicas de Silenciamento de Genes , Genes p16 , Histona-Lisina N-Metiltransferase/metabolismo , Histona-Lisina N-Metiltransferase/fisiologia , Humanos , Lactente , MicroRNAs/fisiologia , Terapia de Alvo Molecular/métodos , Proteína de Leucina Linfoide-Mieloide/metabolismo , Proteína de Leucina Linfoide-Mieloide/fisiologia , Netropsina/farmacologia , Proteínas Nucleares/metabolismo , Regiões Promotoras Genéticas/efeitos dos fármacos , Fatores de Elongação da Transcrição , Regulação para CimaRESUMO
A general route for the semi-automatic synthesis of some new potential minor groove binders was established. Six four-numbered sub-libraries of new netropsin and bis-netropsin analogues have been synthesized using a Syncore Reactor. The structures of the all new substances prepared in this investigation were fully characterized by NMR ((1)H, (13)C), HPLC and LC-MS. The antiproliferative activity of the obtained compounds was tested on MCF-7 breast cancer cells. The ethidium displacement assay using pBR322 confirmed the DNA-binding properties of the new analogues of netropsin and bis-netropsin.
Assuntos
DNA/metabolismo , Netropsina/análogos & derivados , Netropsina/metabolismo , Netropsina/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Técnicas de Química Sintética , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Netropsina/síntese químicaRESUMO
Strain FIM06054 was isolated from a marine sponge sample collected from the East China Sea and was characterised as a strain of Verrucosispora genus on the basis of its 16S rRNA gene sequence. One new compound, FW054-1 (1), together with a known aminofuran compound proximicin A (2), was isolated from the culture broth of Verrucosispora sp. FIM06054. Their structures were elucidated on the basis of spectral analysis. 1 and 2 showed antiproliferative activity against several human tumour cell lines.
Assuntos
Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Micromonosporaceae/química , Netropsina/análogos & derivados , Poríferos/química , Ribose/análogos & derivados , Ribose/isolamento & purificação , Ribose/farmacologia , Animais , Antineoplásicos/química , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Biologia Marinha , Netropsina/química , Netropsina/isolamento & purificação , Netropsina/farmacologia , Ressonância Magnética Nuclear Biomolecular , RNA Ribossômico 16S/genética , Ribose/químicaRESUMO
BACKGROUND: Human malaria parasite infection and its control is a global challenge which is responsible for ~0.65 million deaths every year globally. The emergence of drug resistant malaria parasite is another challenge to fight with malaria. Enormous efforts are being made to identify suitable drug targets in order to develop newer classes of drug. Helicases play crucial roles in DNA metabolism and have been proposed as therapeutic targets for cancer therapy as well as viral and parasitic infections. Genome wide analysis revealed that Plasmodium falciparum possesses UvrD helicase, which is absent in the human host. RESULTS: Recently the biochemical characterization of P. falciparum UvrD helicase revealed that N-terminal UvrD (PfUDN) hydrolyses ATP, translocates in 3' to 5' direction and interacts with MLH to modulate each other's activity. In this follow up study, further characterization of P. falciparum UvrD helicase is presented. Here, we screened the effect of various DNA interacting compounds on the ATPase and helicase activity of PfUDN. This study resulted into the identification of daunorubicin (daunomycin), netropsin, nogalamycin, and ethidium bromide as the potential inhibitor molecules for the biochemical activities of PfUDN with IC50 values ranging from ~3.0 to ~5.0 µM. Interestingly etoposide did not inhibit the ATPase activity but considerable inhibition of unwinding activity was observed at 20 µM. Further study for analyzing the importance of PfUvrD enzyme in parasite growth revealed that PfUvrD is crucial/important for its growth ex-vivo. CONCLUSIONS: As PfUvrD is absent in human hence on the basis of this study we propose PfUvrD as suitable drug target to control malaria. Some of the PfUvrD inhibitors identified in the present study can be utilized to further design novel and specific inhibitor molecules.
Assuntos
Antígenos de Protozoários/metabolismo , DNA Helicases/metabolismo , DNA de Protozoário/metabolismo , Malária Falciparum/metabolismo , Plasmodium falciparum/metabolismo , Células Cultivadas , Daunorrubicina/farmacologia , Etídio/farmacologia , Etoposídeo/farmacologia , Humanos , Malária Falciparum/genética , Terapia de Alvo Molecular , Netropsina/farmacologia , Nogalamicina/farmacologia , Plasmodium falciparum/genética , Plasmodium falciparum/crescimento & desenvolvimento , RNA de Cadeia Dupla/metabolismo , RNA de Protozoário/metabolismoRESUMO
A quick and efficient synthesis and the biological evaluation of promising antitumor-antibiotics proximicins A, B and C are reported. The characteristic repetitive unit of these molecules, the methyl 4-Boc-aminofuran-2-carboxylate 15, was prepared in three synthetic steps in good yield using an optimised copper-catalysed amidation method. The proximicins were evaluated for their antitumor activity using cellular methods. Proximicin B induced apoptosis in both Hodgkin's lymphoma and T-cell leukemia cell lines and proximicin C exhibited significantly high cytotoxicity against glioblastoma and breast carcinoma cells. The proximicins were also screened against Escherichia coli, Enterococcus faecalis and several strains of methicillin-and multidrug-resistant Staphylococcus aureus. Proximicin B showed noteworthy activity against antibiotic-resistant Gram-positive cocci.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Antibióticos Antineoplásicos/síntese química , Antibióticos Antineoplásicos/farmacologia , Netropsina/análogos & derivados , Netropsina/farmacologia , Apoptose/efeitos dos fármacos , Infecções Bacterianas/tratamento farmacológico , Linhagem Celular Tumoral , Enterococcus faecalis/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Feminino , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Neoplasias/tratamento farmacológico , Netropsina/síntese química , Staphylococcus aureus/efeitos dos fármacosRESUMO
Overexpression of high mobility group AT-hook 1 (HMGA1) is common in human cancers. Little is known about the mechanisms underlying its deregulation and downstream targets, and information about its clinical and biological significance in medulloblastoma (MB) is lacking. Here, we demonstrated frequent genomic gain at 6p21.33-6p21.31 with copy number increase leading to overexpression of HMGA1 in MB. The overexpression correlated with a high proliferation index and poor prognosis. Moreover, we found that hsa-miR-124a targeted 3'UTR of HMGA1 and negatively modulated the expression in MB cells, indicating that loss/downregulation of hsa-miR-124a reported in our previous study could contribute to the overexpression. Regarding the biological significance of HMGA1, siRNA knockdown and ectopic expression studies revealed the crucial roles of HMGA1 in controlling MB cell growth and migration/invasion through modulation of apoptosis and formation of filopodia and stress fibers, respectively. Furthermore, we identified cdc25A as a target of HMGA1 and showed that physical interaction between HMGA1 and the cdc25A promoter is required for transcriptional upregulation. In clinical samples, HMGA1 and cdc25A were concordantly overexpressed. Functionally, cdc25A is involved in the HMGA1-mediated control of MB cell growth. Finally, netropsin, which competes with HMGA1 in DNA binding, reduced the expression of cdc25A by suppression of its promoter activity and inhibited in vitro and in vivo intracranial MB cell growth. In conclusion, our results delineate the mechanisms underlying the deregulation and reveal the functional significance of HMGA1 in controlling MB cell growth and migration/invasion. Importantly, the results highlight the therapeutic potential of targeting HMGA1 in MB patients.
Assuntos
Movimento Celular/genética , Proliferação de Células , Neoplasias Cerebelares/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Proteína HMGA1a/metabolismo , Meduloblastoma/metabolismo , Fosfatases cdc25/metabolismo , Citoesqueleto de Actina/metabolismo , Animais , Antivirais/farmacologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Cerebelares/genética , Neoplasias Cerebelares/mortalidade , Neoplasias Cerebelares/patologia , Imunoprecipitação da Cromatina , Aberrações Cromossômicas , Cromossomos Humanos Par 17 , Cromossomos Humanos Par 6 , Relação Dose-Resposta a Droga , Ensaio de Desvio de Mobilidade Eletroforética , Feminino , Citometria de Fluxo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Inativação de Genes , Proteína HMGA1a/genética , Humanos , Masculino , Meduloblastoma/genética , Meduloblastoma/mortalidade , Meduloblastoma/patologia , Camundongos , Camundongos Nus , Invasividade Neoplásica/patologia , Netropsina/farmacologia , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Análise de Sobrevida , Fatores de Tempo , Ensaios Antitumorais Modelo de Xenoenxerto , Fosfatases cdc25/genéticaRESUMO
A compound that can target cells expressing the estrogen receptor (ER), and produce predominantly 3-MeA adducts in those cells has been designed and synthesized. This compound produces mainly the 3-MeA adduct upon reaction with calf thymus DNA, and binds to the ER with a relative binding affinity of 51% (estradiol = 100%). The compound is toxic to ER-expressing MCF-7 breast cancer cells, and pre-treatment with the ER antagonist fulvestrant abrogates the toxicity. Pre-treatment of MCF-7 cells with netropsin, which inhibits N3-adenine methylation by the compound, resulted in a threefold decrease in the toxicity. These results demonstrate the feasibility of this strategy for producing 3-MeA adducts in targeted cells.
Assuntos
Adenina/química , DNA/química , Receptores de Estrogênio/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA/metabolismo , Metilação de DNA , Estradiol/análogos & derivados , Estradiol/farmacologia , Fulvestranto , Humanos , Simulação de Dinâmica Molecular , Netropsina/farmacologia , Ligação Proteica , Receptores de Estrogênio/antagonistas & inibidoresRESUMO
Herein, we study the nanomechanical characteristics of single DNA molecules in the presence of DNA binders, including intercalating agents (ethidium bromide and doxorubicin), a minor groove binder (netropsin) and a typical alkylating damaging agent (cisplatin). We have used magnetic tweezers manipulation techniques, which allow us to measure the contour and persistence lengths together with the bending and torsional properties of DNA. For each drug, the specific variations of the nanomechanical properties induced in the DNA have been compared. We observed that the presence of drugs causes a specific variation in the DNA extension, a shift in the natural twist and a modification of bending dependence on the imposed twist. By introducing a naive model, we have justified an anomalous correlation of torsion data observed in the presence of intercalators. Finally, a data analysis criterion for discriminating between different molecular interactions among DNA and drugs has been suggested.
Assuntos
DNA/química , Antineoplásicos Alquilantes/farmacologia , Fenômenos Biomecânicos , Cisplatino/farmacologia , DNA/efeitos dos fármacos , Doxorrubicina/farmacologia , Etídio/farmacologia , Substâncias Intercalantes/farmacologia , Ligantes , Magnetismo , Netropsina/farmacologia , Conformação de Ácido Nucleico/efeitos dos fármacosRESUMO
DNA-binding small molecules are widespread in the cell and heavily used in biological applications. Here, we use magnetic tweezers, which control the force and torque applied to single DNAs, to study three small molecules: ethidium bromide (EtBr), a well-known intercalator; netropsin, a minor-groove binding anti-microbial drug; and topotecan, a clinically used anti-tumor drug. In the low-force limit in which biologically relevant torques can be accessed (<10 pN), we show that ethidium intercalation lengthens DNA â¼1.5-fold and decreases the persistence length, from which we extract binding constants. Using our control of supercoiling, we measure the decrease in DNA twist per intercalation to be 27.3±1° and demonstrate that ethidium binding delays the accumulation of torsional stress in DNA, likely via direct reduction of the torsional modulus and torque-dependent binding. Furthermore, we observe that EtBr stabilizes the DNA duplex in regimes where bare DNA undergoes structural transitions. In contrast, minor groove binding by netropsin affects neither the contour nor persistence length significantly, yet increases the twist per base of DNA. Finally, we show that topotecan binding has consequences similar to those of EtBr, providing evidence for an intercalative binding mode. These insights into the torsional consequences of ligand binding can help elucidate the effects of small-molecule drugs in the cellular environment.
Assuntos
DNA/química , Anti-Infecciosos/farmacologia , DNA/efeitos dos fármacos , Etídio/farmacologia , Substâncias Intercalantes/farmacologia , Magnetismo , Netropsina/farmacologia , Conformação de Ácido Nucleico/efeitos dos fármacos , Rotação , Inibidores da Topoisomerase I/farmacologia , Topotecan/farmacologia , Torção MecânicaRESUMO
Me-lex(py/py), an adenine-N3-selective alkylating agent, and the reversible minor-groove binder netropsin were used to probe the formation of unusual minor-groove adducts by the cytotoxic hybrid agent PT-ACRAMTU ([PtCl(en)(ACRAMTU)](NO(3))(2); en = ethane-1,2-diamine, ACRAMTU = 1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea). PT-ACRAMTU was found by chemical footprinting to inhibit specific Me-lex-mediated DNA cleavage at several adenine sites but not at nonspecific guanine, which is consistent with the platination of adenine-N3. In a cell proliferation assay, a significant decrease in cytotoxicity was observed for PT-ACRAMTU, when cancer cells were pretreated with netropsin, suggesting that minor-groove adducts in cellular DNA contribute to the biological activity of the hybrid agent.
Assuntos
Adenina/química , Alquilantes/toxicidade , Adutos de DNA/química , Compostos Organoplatínicos/química , Alquilantes/química , Clivagem do DNA/efeitos dos fármacos , Netropsina/farmacologia , Compostos Organoplatínicos/toxicidadeRESUMO
We recently demonstrated that poly(ADP-ribose) polymerase (PARP)-1 is involved in angiogenesis and tumour aggressiveness. In this study we have compared the influence of abrogation of PARP-1 expression by stable gene silencing to that of the pharmacological inhibition of cellular PARP activity using PARP-1/-2 inhibitors on the chemosensitivity of tumour cells to the wide spectrum methylating agent temozolomide (TMZ) and to the N3-adenine selective methylating agent {1-methyl-4-[1-methyl-4-(3-methoxysulfonylpropanamido)pyrrole-2-carboxamido]-pyrrole-2-carboxamido}propane (Me-Lex). Silencing of PARP-1 in melanoma or cervical carcinoma lines enhanced in vitro sensitivity to TMZ and Me- Lex, and induced a higher level of cell accumulation at the G2/M phase of cell cycle with respect to controls. GPI 15427, which inhibits both PARP-1 and PARP-2, increased sensitivity to TMZ and Me-Lex both in PARP-1-proficient and - deficient cells. However, it induced different cell cycle modulations depending on PARP-1 expression, provoking a G2/M arrest only in PARP-1 silenced cells. Treatment of PARP-1 silenced cells with TMZ or Me-Lex resulted in a more extensive phosphorylation of Chk-1 and p53 as compared to PARP-1 proficient cells. The combination of the methylating agents with GPI 15427 increased Chk-1 and p53 phosphorylation both in PARP-1 proficient or deficient cells. When mice challenged with PARP-1 silenced melanoma cells were treated with the TMZ and PARP inhibitor combination there was an additional reduction in tumour growth with respect to treatment with TMZ alone. These results suggest the involvement of PARP-2 or other PARPs, in the repair of DNA damage provoked by methylating agents, highlighting the importance of targeting both PARP-1 and PARP-2 for cancer therapy.
Assuntos
Antineoplásicos/farmacologia , Dacarbazina/análogos & derivados , Inibidores Enzimáticos/farmacologia , Netropsina/análogos & derivados , Inibidores de Poli(ADP-Ribose) Polimerases , Animais , Western Blotting , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Dacarbazina/farmacologia , Sinergismo Farmacológico , Citometria de Fluxo , Fase G2/efeitos dos fármacos , Células HeLa , Humanos , Melanoma Experimental/patologia , Metilação , Camundongos , Netropsina/farmacologia , Poli(ADP-Ribose) Polimerases/genética , TemozolomidaRESUMO
The synthesis and in vitro anti-tumor 60 cell lines screen of a novel series of anthracenyl isoxazole amides (AIMs) (While not a strict acronym, the designation AIM is in honor of the memory of Professor Albert I. Meyers.) (22-33) are described. The molecules consist of an isoxazole that pre-organizes a planar aromatic moiety and a simple amide and/or lexitropsin-oligopeptide. The new conjugate molecules were prepared via doubly activated amidation modification of Weinreb's amide formation technique, using SmCl(3) as an activating agent which produces improved yields for sterically hindered 3-aryl-4-isoxazolecarboxylic esters. The results of the National Cancer Institute's (NCI) 60 cell line screening assay show a distinct structure activity relationship (SAR), wherein a trend of the highest activity for molecules with one N-methylpyrrole peptide. Evidence consistent with a mechanism of action via the interaction of these compounds with G-quadruplex (G4) DNA and a structural based rational for the observed selectivity of the AIMs for G4 over B-DNA is presented.
Assuntos
Antracenos/síntese química , Antracenos/farmacologia , Antineoplásicos/síntese química , Azóis/síntese química , Azóis/farmacologia , Netropsina/análogos & derivados , Antracenos/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Azóis/química , Linhagem Celular Tumoral , Desenho de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Espectroscopia de Ressonância Magnética , Netropsina/síntese química , Netropsina/química , Netropsina/farmacologia , Espectrometria de Massas por Ionização por Electrospray , Relação Estrutura-AtividadeRESUMO
A family of three novel aminofuran antibiotics named as proximicins was isolated from the marine Verrucosispora strain MG-37. Proximicin A was detected in parallel in the marine abyssomicin producer "Verrucosispora maris" AB-18-032. The characteristic structural element of proximicins is 4-amino-furan-2-carboxylic acid, a hitherto unknown gamma-amino acid. Proximicins show a weak antibacterial activity but a strong cytostatic effect to various human tumor cell lines.
Assuntos
Actinobacteria/metabolismo , Antibacterianos/farmacologia , Antibióticos Antineoplásicos/farmacologia , Netropsina/análogos & derivados , Actinobacteria/química , Actinobacteria/classificação , Antibacterianos/biossíntese , Antibacterianos/isolamento & purificação , Antibióticos Antineoplásicos/biossíntese , Antibióticos Antineoplásicos/isolamento & purificação , Bactérias/efeitos dos fármacos , Linhagem Celular Tumoral , Fenômenos Químicos , Físico-Química , Cromatografia Líquida de Alta Pressão , Fermentação , Humanos , Testes de Sensibilidade Microbiana , Netropsina/biossíntese , Netropsina/isolamento & purificação , Netropsina/farmacologia , Espectrofotometria UltravioletaAssuntos
Antineoplásicos/farmacologia , Inibidor de Quinase Dependente de Ciclina p21/genética , Netropsina/análogos & derivados , Netropsina/farmacologia , Proteína Supressora de Tumor p53/genética , Regulação para Cima/efeitos dos fármacos , Actinobacteria/química , Furanos , Estrutura Molecular , Netropsina/química , Netropsina/isolamento & purificaçãoRESUMO
The design of small synthetic molecules that can be used to affect gene expression is an area of active interest for development of agents in therapeutic and biotechnology applications. Many compounds that target the minor groove in AT sequences in DNA are well characterized and are promising reagents for use as modulators of protein-DNA complexes. The mammalian high-mobility-group transcriptional factor HMGA2 also targets the DNA minor groove and plays critical roles in disease processes from cancer to obesity. Biosensor-surface plasmon resonance methods were used to monitor HMGA2 binding to target sites on immobilized DNA, and a competition assay for inhibition of the HMGA2-DNA complex was designed. HMGA2 binds strongly to the DNA through AT hook domains with KD values of 20-40 nM depending on the DNA sequence. The well-characterized minor groove binder netropsin was used to develop and test the assay. The compound has two binding sites in the protein-DNA interaction sequence, and this provides an advantage for inhibition. An equation for analysis of results when the inhibitor has two binding sites in the biopolymer recognition surface is presented with the results. The assay provides a platform for discovery of HMGA2 inhibitors.