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IL-6-engineered DC stimulate efficient antitumor immunity via enhanced and prolonged T cell cytotoxicity and survivalZhang, Bei 06 March 2009
Dendritic cells (DCs) modified by some immunomodulatory genes can stimulate a strong antitumor immunity and improve the treatment of tumor cells on the condition that the sources of tumor-associated antigens (TAAs) are available. IL-6, a pleotropic cytokine, has been found to inhibit CD4+25+ regulatory T (Treg)-cell-mediated immune suppression and decrease activation-induced cell death (AICD) without interfering the process of T-cell activation. To enhance DC-based cancer vaccine, we engineered DCs to express transgene IL-6.<p>
We constructed a fiber-modified recombinant adenovirus vector AdVIL-6 expressing IL-6, infected DCs with AdVIL-6, and then investigated the efficacy of antitumor immunity induced by vaccination with DCs engineered to express IL-6 transgene. We demonstrated that DCs infected with the recombinant adenovirus AdVIL-6 induced DC maturation by up-regulation of the expression of MHC class U (Iab), CD40, CD54 and CD80 expression. We also demonstrated that vaccination of OVA-pulsed AdVIL-6-infected DCs (DCOVA/AdVIL-6) was able to stimulate a stronger OVA-specific effector CD8+ cytotoxic T lymphocyte (CTL) response than vaccination with the control virus AdVpLpA-infected DCs (DCOVA/AdVpLpA). More importantly, vaccination of mice with DCOVA/AdVpLpA could protect 100% mice from intravenous (i.v.) challenge of a low dose (0.5~105 cells per mouse, 8/8 mice protected) of OVA-expressing BL6-10OVA tumor cells, but only 63% mice from i.v. challenge of a high dose (1~105 cells per mouse, 5/8 mice protected) of BL6-10OVA tumor cells. However, vaccination of DCOVA/AdVIL-6 induced an augmented antitumor immunity in vivo by complete protection of mice (8/8) from challenge of both low and high doses of BL6-10OVA tumor cells.<p>
To study the immune mechanism underlying the result of IL-6 engineered-DC vaccine, we generated the DCOVA/AdVIL-6-activated OTI CD8+ T cells and DCOVA/AdVpLpA-activated OTI CD8+ T cells. We demonstrated that DCOVA/AdVIL-6-activated CD8+ T cells displayed a higher level of CD62L, FasL and perforin than DCOVA/AdVpLpA-activated CD8+ T cells. DCOVA/AdVIL-6-activated CD8+ T cells had a prolonged T cell survival after they were transferred into C57BL/6 mice. Furthermore, the results of the animal study showed that 100% of mice bearing OVA-expressing EG7 tumors (8mm in diameter, 8 mice per group) were tumor-free after they were i.v. treated with DCOVA/AdVIL-6-activated CD8+ T cells (2~106 cells per mouse). However, the control DCOVA/AdVpLpA-activated CD8+ T cells failed in eradication of EG7 tumors in all 8/8 mice.<p>
Taken together, Adenovirus-mediated IL-6 transgene engineered DC vaccine stimulates efficient CD8+ T cell responses and antitumor immunity via enhanced T cell cytotoxicity and prolonged T cell survival. DCs engineered to express IL-6 by adenovirus-mediated IL-6 gene transfer may offer a new strategy in production of DC cancer vaccines.
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IL-6-engineered DC stimulate efficient antitumor immunity via enhanced and prolonged T cell cytotoxicity and survivalZhang, Bei 06 March 2009 (has links)
Dendritic cells (DCs) modified by some immunomodulatory genes can stimulate a strong antitumor immunity and improve the treatment of tumor cells on the condition that the sources of tumor-associated antigens (TAAs) are available. IL-6, a pleotropic cytokine, has been found to inhibit CD4+25+ regulatory T (Treg)-cell-mediated immune suppression and decrease activation-induced cell death (AICD) without interfering the process of T-cell activation. To enhance DC-based cancer vaccine, we engineered DCs to express transgene IL-6.<p>
We constructed a fiber-modified recombinant adenovirus vector AdVIL-6 expressing IL-6, infected DCs with AdVIL-6, and then investigated the efficacy of antitumor immunity induced by vaccination with DCs engineered to express IL-6 transgene. We demonstrated that DCs infected with the recombinant adenovirus AdVIL-6 induced DC maturation by up-regulation of the expression of MHC class U (Iab), CD40, CD54 and CD80 expression. We also demonstrated that vaccination of OVA-pulsed AdVIL-6-infected DCs (DCOVA/AdVIL-6) was able to stimulate a stronger OVA-specific effector CD8+ cytotoxic T lymphocyte (CTL) response than vaccination with the control virus AdVpLpA-infected DCs (DCOVA/AdVpLpA). More importantly, vaccination of mice with DCOVA/AdVpLpA could protect 100% mice from intravenous (i.v.) challenge of a low dose (0.5~105 cells per mouse, 8/8 mice protected) of OVA-expressing BL6-10OVA tumor cells, but only 63% mice from i.v. challenge of a high dose (1~105 cells per mouse, 5/8 mice protected) of BL6-10OVA tumor cells. However, vaccination of DCOVA/AdVIL-6 induced an augmented antitumor immunity in vivo by complete protection of mice (8/8) from challenge of both low and high doses of BL6-10OVA tumor cells.<p>
To study the immune mechanism underlying the result of IL-6 engineered-DC vaccine, we generated the DCOVA/AdVIL-6-activated OTI CD8+ T cells and DCOVA/AdVpLpA-activated OTI CD8+ T cells. We demonstrated that DCOVA/AdVIL-6-activated CD8+ T cells displayed a higher level of CD62L, FasL and perforin than DCOVA/AdVpLpA-activated CD8+ T cells. DCOVA/AdVIL-6-activated CD8+ T cells had a prolonged T cell survival after they were transferred into C57BL/6 mice. Furthermore, the results of the animal study showed that 100% of mice bearing OVA-expressing EG7 tumors (8mm in diameter, 8 mice per group) were tumor-free after they were i.v. treated with DCOVA/AdVIL-6-activated CD8+ T cells (2~106 cells per mouse). However, the control DCOVA/AdVpLpA-activated CD8+ T cells failed in eradication of EG7 tumors in all 8/8 mice.<p>
Taken together, Adenovirus-mediated IL-6 transgene engineered DC vaccine stimulates efficient CD8+ T cell responses and antitumor immunity via enhanced T cell cytotoxicity and prolonged T cell survival. DCs engineered to express IL-6 by adenovirus-mediated IL-6 gene transfer may offer a new strategy in production of DC cancer vaccines.
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AvaliaÃÃo do potencial antitumoral dos hidrobenzofuranÃides isolados das folhas da Tapirira guianensis (anacardiaceae) / Antitumor potential of hydrobenzofuranoids isolated from the leaves of tapirira guianensis (anacardiaceae)PatrÃcia MarÃal da Costa 12 December 2006 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Os hidrobenzofuranÃides obtidos da Tapirira guianensis sÃo derivados alquilados da ciclohexanona, que parecem ser possÃveis precursores dos lipÃdios fenÃlicos. O presente trabalho avaliou, inicialmente, a atividade dos nove hidribenzofuranÃides em linhagens de cÃlulas, onde a amostra SJC-8 mostrou a citotoxicidade mais elevada. Posteriormente, foram avaliados os possÃveis mecanismos pelo qual esta amostra desenvolve seu efeito citotÃxico. No teste de MTT em painel de linhagens adicionais a SJC-8 apresentou valores de CI50 variando de 0.3 a 6.2Âg/mL. No teste de toxicidade aguda em nÃuplios de artÃmia e de atividade hemolÃtica em eritrÃcitos de camundongos, a SJC-8 nÃo desenvolveu toxicidade e hemÃlise, respectivamente. O mecanismo de aÃÃo da SJC-8 foi, entÃo, estudado. A viabilidade das cÃlulas HL-60 foi afetada pela SJC-8 apÃs um perÃodo de exposiÃÃo de 24h, quando analisada por exclusÃo por azul de tripan. Nas menores concentraÃÃes nÃo houve aumento do nÃmero de cÃlulas nÃo-viÃveis, mas apenas uma reduÃÃo da proliferaÃÃo celular (aÃÃo citostÃtica), enquanto que nas duas maiores concentraÃÃes, houve reduÃÃo do nÃmero de cÃlulas viÃveis e aumento do nÃmero de cÃlulas nÃo-viÃveis (efeito citotÃxico), o que corrobora com os achados da analise morfolÃgica, onde observou-se um aumento do nÃmero de cÃlulas mortas. A atividade citotÃxica da SJC-8 està relacionada com a inibiÃÃo da sÃntese de DNA, como revelado pela incorporaÃÃo do BrdU, alÃm de poder estar envolvida com a inibiÃÃo da Topoisomerase 1. Submetida ao estudo de toxicogenÃtica pelo teste do cometa em HL-60, a SJC-8 elevou os Ãndices e freqÃÃncias de dano de maneira concentraÃÃo-dependente, sendo observados tipos de danos maiores nas concentraÃÃes mais elevadas. A administraÃÃo de SJC-8 (25 ou 50mg/kg/dia) inibiu o desenvolvimento de tumor sÃlido em camundongos transplantados com Sarcoma 180 em 12,3 e 59,8% respectivamente. A atividade antitumoral da SJC-8 està relacionada com a inibiÃÃo da proliferaÃÃo do tumor. A anÃlise histopatolÃgica mostrou de forma reversÃvel, que o fÃgado à o alvo de toxicidade da droga. De fato, a atividade antitumoral da SJC-8 esta relacionada com um efeito antiproliferativo direto nas cÃlulas tumorais, sendo possÃvel assim que esta amostra possa atuar como possÃvel protÃtipo de novos agentes antitumorais. / The hydrobenzofuranoids obtained from Tapirira guianensis are alkylated derivates of cyclo-hexanone, which appear to be precursors of phenolic lipids. The present study initially examined the activity of nine hydrobenzofuranoids in cell lines, where the compound SJC-8 showed the highest cytotoxicity. In later studies, the cytotoxicity of this sample was investigated with regard to the possible mechanism of action. In the MTT assay, SJC-8 showed IC50 values of 0.3 to 6.2Âg/mL in a panel of cell lines. In acute toxicity assays in artemia nauplii and hemolytic activity in mouse erythrocytes, SJC-8 did not demonstrate any toxicity or hemolysis, respectively. The mechanism of action of SJC-8 was then studied. SJC-8 affected cell viability in HL-60 after an exposure period of 24h, when determined by trypan blue exclusion. At lower concentrations, there was no increase in the number of non-viable cells but only a reduction in cell proliferation (cytostatic effect). However, at the two highest concentrations, there was a decrease in the number of viable cells and increase in number of non-viable cells (cytotoxic effect), which corroborate the findings of morphologic analysis showing an increase in the number of dead cells. The cytotoxicity of SJC-8 involves the inhibition of DNA synthesis, as revealed by inhibition of BrdU incorporation into DNA and of topoisomerase 1 activity. SJC-8 was tested for genotoxicity using the comet assay in HL-60 cells, and was found to cause an increase in the frequency of DNA damage in a concentration-dependent manner, where more severe damage was seen at higher concentrations of SJC-8. The administration of SJC-8 (25 or 50 mg/kg/day) inhibited solid tumor growth in mice transplanted with sarcoma 180, by 12.3 and 59.8%, respectively. The antitumor activity of SJC-8 is attributed to inhibition of tumor cell proliferation. Histopathologic analysis showed in a reversible manner that the liver is the target of drug toxicity. In conclusion, SJC-8 has antitumor activity where it has a direct antiproliferative effect on tumor cells, and may therefore serve as a prototype for new antitumor agents.
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STAT2 in the regulation of tumor growth and antitumor effects of type I interferons in a mouse model of melanomaYue, Chanyu January 2013 (has links)
Signal Transducer and Activator of Transcription (STAT) 2 is one of seven members of the STAT family of transcription factors with dual roles in signal transduction and gene activation. STAT2 is a central transcription factor that regulates the antiviral, apoptotic and cell growth inhibitory effects of type I interferons (IFN-α/β), a small family of secreted glycoproteins induced by the host after sensing the presence of tumor cells and pathogens. The creation of Stat2-/- mice established the pivotal role of STAT2 in type I IFN signaling and in antiviral immunity. In vitro studies conducted in STAT2 deficient tumor cell lines suggested a role in suppressing tumor cell growth in response to IFN treatment. Based on these properties STAT2 is presumed to have tumor suppressor functions but data to support this notion in animal models of cancer are limited. To address the role of STAT2 in cancer, I used the murine B16-F1 tumor transplantation model of human melanoma. The B16-F1 melanoma cell line was established from a spontaneous tumor that arose in mice. I discovered that tumor cells transplanted subcutaneously in Stat2-/- mice grew more aggressively than in the counterpart wild type mice. Closer examination of B16-F1 tumors harvested from wild type and Stat2-/- mice revealed an unexpected dramatic similar reduction of STAT2 and STAT1 proteins. Yet soluble factors secreted by B16-F1 tumors established in Stat2-/- mice alone were sufficient to enhance proliferation of B16-F1 tumor cells. I further showed that tumor-bearing wild type mice treated with IFN-β developed smaller tumors compared to Stat2-/- mice, whose tumors continued to grow and hence were unresponsive to IFN intervention. Lastly, to elucidate a mechanism that leads to enhanced tumor growth in Stat2-/- mice, I questioned the involvement of the host immune response in restricting tumor growth. I found that tumor specific T cell priming by Stat2-/- dendritic cells (DCs) was defective since generated cytotoxic T cells (CD8+ T lymphocytes) produced low levels of IFN-γ and IL-2 and adoptive transfer of these B16-F1 tumor specific CD8+ T cells in B16-F1 bearing Stat2-/- mice did not cause tumor regression with IFN-β intervention. Collectively, my findings reveal that host STAT2 restricts the establishment of melanoma tumors. More importantly, type I IFN/STAT2 signaling on DCs plays a pivotal role in tumor antigen cross-presentation to CD8+ T cells and in the development of a protective antitumor response resulting in tumor rejection. To now address whether STAT2 expression in cancer cells could influence tumor establishment and the antitumor effects of type I IFNs, STAT2 expression was silenced in B16-F1 tumor cells. Contrary to my expectation, silencing STAT2 augmented the growth inhibitory effects of IFN-β both in vitro and in vivo. However, loss of STAT2 expression in the tumor did not cause B16-F1 tumor cells to grow more aggressively compared to control B16-F1 cells. Furthermore, compared to B16-F1 control cells, STAT2-silenced B16-F1 cells showed an initial delay but later persistent STAT activation and formation of the ISGF3 transcriptional complex (consisting of STAT1, STAT2 and IRF9). This observation paralleled with an initial delay and then later an increase in the expression of IFN regulated genes. In addition, reduced activation of STAT5 induced by IFN-β was observed in STAT2-silenced B16-F1 cells. This may partially explain the enhanced growth inhibitory effects of type I IFNs. Together these results shed light on the unexpected role of tumor STAT2 expression in diminishing the efficacy of type I IFN treatment of melanoma. / Biochemistry
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Development of tumour selective and endoprotease-activated anticancer therapeutics.Gill, Jason H., Loadman, Paul January 2008 (has links)
No
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Avaliação do efeito antiproliferativo da apocinina e diapocinina em células de osteossarcoma humano / Evaluation of the antiproliferative effect of apocynin and diapocynin on human osteosarcoma cellsMatos, Adriana Arruda 24 January 2018 (has links)
O osteossarcoma (OS) é o tumor maligno primário mais comum do tecido ósseo, caracterizado pela formação de osteócitos anormais. Apesar do avanço nas terapias convencionais (quimioterapia e retirada do tumor), essas não conseguem eliminar totalmente as células tumorais e impedir a progressão da doença. Recentemente, agentes derivados de fontes naturais ganharam considerável atenção por causa de sua segurança, eficácia e disponibilidade imediata. Nesse sentido, a apocinina, inibidor do complexo NADPH-oxidase, vem sendo estudada como agente antitumoral em alguns tipos de câncer como: pâncreas, próstata, pulmão e mama. Apocinina é um pró-fármaco e sua ação parece estar relacionada à sua conversão produzindo a diapocinina, a qual se mostrou mais efetiva do que a apocinina. Portanto, o objetivo desse estudo é avaliar, in vitro, o potencial antitumoral da apocinina e diapocinina em células de osteossarcoma humano. Para isso, foram utilizados osteoblastos humanos normais (HOb) e osteossarcoma humano imortalizadas (SaOS-2) tratados ou não com apocinina e diapocinina em diversas concentrações. Foram realizados os ensaios de viabilidade celular, alterações morfológicas, apoptose celular, produção de espécies reativas de oxigênio (EROs), formação de colônias, migração, invasão e expressão do fator indutor de hipóxia-1alfa (HIF-1). Também foram conduzidos ensaios para verificar a atividade de metaloproteinase de matriz (MMP) 2 e 9. Os resultados em SaOS-2 mostraram que o tratamento com apocinina nas concentrações de 1,5 e 3 mM; e diapocinina nas concentrações de 0,75 e 1,5 mM reduziram a viabilidade; aumentaram o número de células em apoptose e diminuíram a produção de EROs; sem causar danos às células HOb. Além disso, essas mesmas concentrações inibiram a migração e invasão celular; diminuíram a expressão de HIF-1; e reduziram a atividade de MMP-2 em SaOS-2. Considerando os resultados obtidos, concluímos que a apocinina e diapocinina podem atuar como possíveis moduladores de células tumorais, sendo que a diapocinina mostrou ser mais efetiva nos parâmetros testados. / Osteosarcoma (OS) is the most common primary malignant tumor of bone tissue, characterized by the formation of abnormal osteocytes. Despite advances in conventional therapies (chemotherapy and surgery) they cannot completely eliminate tumor cells and prevent the progression of the disease. Recently, agents derived from natural sources have achieved considerable attention because of their safety, efficacy and immediate availability of therapies. In this way, apocynin, an inhibitor of the NADPH-oxidase complex, has been studied as an antitumor agent in some types of cancer, such as pancreas, prostate, lung and breast. Apocynin is a prodrug and its action indicate to be related to its conversion to diapocynin, which has been shown to be more efficient than apocynin itself. Thus, the aim of this study is to evaluate, in vitro, the antitumor potential of apocynin and diapocynin in human osteosarcoma cells. For this, normal human osteoblasts (HOb) and immortalized human osteosarcoma cells (SaOS-2) were treated or no-treated with apocynin and diapocynin in various concentrations. Cell viability assay, morphological alterations, cellular apoptosis, reactive oxygen species (ROS) production, colony formation, migration, invasion and expression of hypoxia-inducible factor-1 alpha (HIF-1) were performed. We also performed assays to verify the activity of matrix metalloproteinase (MMP) 2 and 9. The results in SaOS-2 showed that treatment with apocynin at concentrations of 1,5 e 3 mM; and diapocynin at concentrations of 0,75 e 1,5 mM reduced cell viability; increased the number of cells in apoptosis and decreased the production of ROS; without damaging HOb cells. Moreover, these same concentrations inhibited cell migration and invasion; decreased HIF-1 expression; and reduced MMP 2 activity in SaOS-2. Considering the results, we suggest that apocynin and diapocynin may act as possible modulators of tumor cells, and diapocynin has been shown to be more effective.
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Synthesis and characterization of new metal complexes with antitumor propertyAlrashdi, Salma 01 July 2016 (has links)
Recent studies of new metal complexes for antitumor properties have focused on the speeding up of the treatment process in chemotherapy. In this study, focus was put on the two new metal complexes, copper (LCu(OAc)2) and iron {LFeCl2[FeCl4]}complexes that have been synthesized. Both new complexes have been characterized by FT-IR and UV-Vis. In addition, the structures of two complexes have been established by the X-ray crystallography. The molecular structures of (LCu(OAc)2) and LFeCl2[FeCl4] have been determined confirming that the ligand is bidentate (N,N) imidazopyridine and adopted octahedral configurations. The two complexes and ligand L have been tested for biological activity against prostate cancer cell line LNCaP. The complexes have shown promising results for the anti-proliferative activity. Although the ligand displayed high cytotoxicity toward prostate cancer cell line, the complexes have shown far better cytotoxicity property than the ligand.
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Antitumour MetallocenesMokdsi, George January 2000 (has links)
This thesis reports a study of the chemical stability and coordination chemistry of several antitumour metallocenes Cp2MCl2 (Cp = h5-C5H5; M = Ti 1, V 2, Nb 3, Mo 4), as well as derivatives of Cp2TiCl2 1, with nucleic acids, nucleic acid constituents and proteins. These studies were carried out in order to identify the biologically active species and more fully understand the molecular level mechanism of action of the antitumour metallocenes, in particular Cp2TiCl2 1, which is currently undergoing phase II clinical trials. The interactions of Cp2MoCl2 4 with four oligonucleotides were studied by 1H and 31P NMR spectroscopy. In 50 mM salt solutions of Cp2MoCl2 4, hydrolysis of the halide ligands occurred to give a solution with pD -2, containing a species in which both Cp rings remain metal bound for 24 h. At pD -7, partial hydrolysis of the Cp rings (-30percent) occurred after 24 h. Addition of an aqueous solution of Cp2MoCl2 4 in 50 mM salt to the self-complementary sequence d(CGCATATGCG)2, maintaining the pD at 6.0-7.0, showed no evidence for the formation of a metallocene-oligonucleotide complex and only peaks arising from hydrolysis of Cp2MoCl2 4 were detected. A similar result was obtained in titration experiments with the single stranded sequence d(ATGGTA) at pD 6.5-7.0. However, at pD 3.0, new signals assigned to a molybdocene-oligonucleotide complex(es), which was stable for hours at pD 3.0, were detected; while at pD -7 the complex is destabilised and only peaks arising from hydrolysis of Cp2MoCl2 4 were detected. Titration experiments at low pD with Cp2MoCl2 4 and the dinucleotide dCG were consistent with formation of a complex arising due to coordination of molybdenum to guanine N7 and/or cytosine N3. The results obtained showed that stable oligonucleotide adducts were not formed in 50 mM salt at pD -7 and hence it is highly unlikely that formation of molybdocene-DNA adducts in vivo is the primary action that is responsible for the antitumour properties of Cp2MoCl2 4. The rate of hydrolysis of the aromatic rings of Cp2TiX2 (X equals Cl 1, OCOCH2NH3Cl 27) and the dimethylsubstituted derivatives (MeCp)2TiX2 (X equals Cl 34, OCOCH2NH3Cl 41), in aqueous solutions at pD 2-8 was studied by 1H NMR spectroscopy. Rapid hydrolysis of both the halide/glycine and Cp ligands in Cp2TiX2 (X equals Cl 1, OCOCH2NH3Cl 27) occurred and predominantly gave a precipitate at pD -7. In contrast, under the same experimental conditions, the predominant species present in aqueous solutions of (MeCp)2TiX2 (X equals Cl 34, OCOCH2NH3Cl 41) at pH 2-8 contained both MeCp rings metal bound. At pD < 5, Cp2TiX2 (X equals Cl 1, OCOCH2NH3Cl 27) and (MeCp)2TiX2 (X equals Cl 34, OCOCH2NH3Cl 41) formed similar complex(es) with purine nucleotides. However, at pD >5, stable adducts between nucleotides and Cp2TiX2 (X equals Cl 1, OCOCH2NH3Cl 27) were not formed. In contrast, (MeCp)2TiX2 (X equals Cl 34, OCOCH2NH3Cl 41) formed complex(es) with 5'-dAMP or 5'-dGMP, which were stable for 24 h. These results suggest that formation of stable chelates between (MeCp)2TiX2 (X equals Cl 34, OCOCH2NH3Cl 41) and nucleic acid constituents in vivo is possible. However, the methyl substituted derivatives 34 and 41 did not show any antitumour activity against EAT in mice when administered in either 10percentDMSO/90percentsaline or in water at pH 6.2-6.4, which suggests that the labile Cp-Ti bond present in Cp2TiCl2 1 is required for antitumour activity. The synthesis of a range of Cp substituted titanocene derivatives was investigated in an attempt to prepare derivatives with modified Cp stability in comparison to the methyl substituted derivatives. The synthesis of derivatives (CpCH2Y)2TiCl2 where Y equals ?CHO 43, ?CONMe2 44, ?NO2 45, (RCp)2TiCl2 where R equals ?COMe 46, ?COOMe 47 or ?CONMe2 48, (CpNMe2)2TiCl2 62 and (Cp(CH2)2NMe2)2TiCl2 63 was unsuccessful, due to the presence of coordinating substituents on the Cp rings and poor stability in polar, protic solvents. Hence, these derivatives were excluded from further studies. The rate of hydrolysis of the Cp rings of Cp2TiX2 (X equals Cl 1, OCOCCl3 22 and OCOCH2NH3Cl 27) in aqueous solutions, 10percentDMSO/90percentD2O and 100percent DMSO was monitored by 1H NMR spectroscopy. Rapid hydrolysis of both the carboxylate and Cp ligands of Cp2TiX2 (OCOCCl3 22 and OCOCH2NH3Cl 27) occurred in DMSO to give biologically inactive species. The rate of these reactions were concentration dependent as dilution of these samples with saline or water to give the therapeutic conditions of 10percentDMSO/90percentD2O slowed the hydrolysis chemistry. In contrast, samples of Cp2TiX2 (X equals Cl 1 and OCOCH2NH3Cl 27) dissolved in water, gave solutions containing the presumed antitumour active species in which the halide or glycine ligands have been hydrolysed but the Cp rings remain metal bound. Thus, charged X ligands may be incorporated into Cp2TiX2 and will give comparable activity to Cp2TiCl2 1 provided the samples are administered in water. The antitumour metallocenes Cp2MCl2 (M equals Ti 1, V 2, Nb 3, Mo 4) and the inactive derivative (MeCp)2TiCl2 34 were found to inhibit the relaxation of supercoiled plasmid DNA pBR322 by human topoisomerase II in vitro. These results implicated the inhibition of topoisomerase II in the mechanism of antitumour activity although there was no direct correlation between the in vitro results with biological activity against EAT in vivo. UV spectroscopy confirmed that the metallocenes Cp2MCl2 (M equals Ti 1, Mo 4) became associated with and were stabilised to hydrolysis by calf thymus DNA but not with human serum albumin. ICP-AES was used to measure the amount of metal associated with either DNA or human serum albumin after incubation with Cp2MCl2 (M equals Ti 1, Nb 3, Mo 4) and dialysis of these solution. The results confirmed that DNA stabilises or becomes associated with the metallocenes. However, errors associated with the ICP-AES measurements did not allow these results to be quantified. 1H NMR spectroscopy was used to show that the antitumour metallocene Cp2MoCl2 4 formed an adduct with glutathione 72 in the pH range 3-7 through the sulfur donor group. In comparison, the antitumour metallocenes Cp2MCl2 (M equals Ti 1, Nb 3) showed limited adduct formation with glutathione 72 at pH -3 and no adducts were detected at pH > 5.5.
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Infrared spectroscopy as a new tool for the screening of antitumoral agents inducing original therapeutic action. La spectroscopie infrarouge comme outil de screening pour l’identification de nouveaux agents thérapeutiquesGasper, Régis 26 November 2010 (has links)
Actuellement le criblage en vue de la recherche de nouveaux agents antitumoraux se base principalement sur la qualité cytotoxique d’une molécule. Le principal défaut de cette approche est qu’aucune sélection n’est faite sur le mode d’action du médicament. L’objectif de ce travail est la mise au point d’une méthode permettant un classement rapide et objectif du mode d’action de molécules à visée thérapeutique par spectroscopie infrarouge.
La spectroscopie infrarouge est une technique d’absorption de la lumière fournit la signature chimique d’un échantillon. L’excellente qualité du signal rend possible son utilisation comme outil discriminant. En outre, cette technique d’analyse se démarque des autres par son caractère non destructif et la rapidité d’acquisition des données. Elle se révèlerait donc une méthode de choix pour effectuer du criblage de molécules en vue de la recherche de nouveaux agents thérapeutiques.
Dans un premier temps nous avons voulu évaluer la possibilité d’utiliser la spectroscopie infrarouge pour isoler la signature spectrale du mode d’action induit par des concentrations sub-létales de ouabaïne, un composé de la famille des cardénolides, sur une lignée tumorale de prostate. Nous avons montré que cette signature évolue au cours du temps et peut-être corrélée aux données biologiques décrites dans la littérature. Nous avons également mis en évidence pour la première fois une modification de la composition lipidique de la cellule. Cette altération a été caractérisée au cours du temps par spectrométrie de masse.
Nous avons ensuite voulu définir les limites de la méthode. La littérature souligne la diversité des modes d’action que peut induire un agent thérapeutique selon sa concentration. Nous avons montré que cette diversité se reflète sur le spectre infrarouge de cellules tumorales traitées à la ouabaïne en distinguant au moins deux modes d’action distincts, dépendant de la concentration en ouabaïne. Par ailleurs, nous avons montré que la confluence pouvait modifier significativement le spectre infrarouge d’une cellule. Neanmoins cette signature est unique et orthogonale à celle induite par la ouabaïne.
Finalement, nous avons évalué le potentiel de la spectroscopie infrarouge à distinguer des modes d’action induits par des molécules à la structure chimique proche. Nous avons montré qu’il était possible de caractériser spécifiquement chacun des modes d’action. D’autre part nous avons mis en évidence que les modes d’action de molécules issues d’une même classe d’agent thérapeutique conduisaient à des signatures spectrales similaires. Cette partie du travail souligne la possibilité d’utilisation de la spectroscopie infrarouge pour un classement objectif, uniquement basé sur leur mode d’action d’agents thérapeutiques potentiels.
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Development of chemotherapies for hormone-dependent breast and prostate cancersMorrow, Michael Derek 17 February 2005 (has links)
Cancer is a leading cause of human mortality worldwide, and is expected to soon become the overall leading cause of death in the United States. Some cancers are hormone-related, including the sex-specific cancers of the breast (predominantly in women) and prostate (in men). In both cases, early stage tumors are responsive to inhibitory endocrine-based therapies. However, both cancers progress to hormone-nonresponsive states and this is in part due to altered properties of the primary nuclear hormone receptor signaling pathway (estrogen receptor [ER] in breast; androgen receptor [AR] in prostate). Other nuclear receptors are thus being investigated as therapeutic targets due to their crosstalk with hormone receptor pathways and these include the aryl hydrocarbon receptor (AhR), peroxisome proliferator activated receptor γ(PPARγ, retinoic acid receptor and retinoid X receptor (RAR/RXR), and vitamin D receptor (VDR). Previous studies have demonstrated that the AhR mediates chemoprotective, antiestrogenic, and tumoristatic effects in experimental models, and relatively non-toxic selective aryl hydrocarbon receptor modulators (SAhRMs) have been developed. Studies in this dissertation have investigated the therapeutic properties of a new class of compounds related to the SAhRM 3,3‘-diindolylmethane (DIM) in models of breast cancer. Additionally, the potential therapeutic role of the AhR in human prostate cancer cells has been investigated. Several ring- and methylene-substituted DIMs exhibited antiestrogenic and tumoristatic activities in breast cancer cells and in carcinogen-induced rat mammary tumors. At least some of the methylene-substituted DIMs act through PPARγ. The AhR is expressed in LNCaP and iv
22Rv1 prostate cancer cells and AhR agonists inhibit cell growth and AR-induced transactivation through pathways independent of androgen receptor downregulation.
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