WO2013053034A2 - Thiazacridines used in anti-cancer therapy - Google Patents
Thiazacridines used in anti-cancer therapy Download PDFInfo
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- WO2013053034A2 WO2013053034A2 PCT/BR2012/000421 BR2012000421W WO2013053034A2 WO 2013053034 A2 WO2013053034 A2 WO 2013053034A2 BR 2012000421 W BR2012000421 W BR 2012000421W WO 2013053034 A2 WO2013053034 A2 WO 2013053034A2
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- C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
- C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
- C07D277/08—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
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- A61P35/00—Antineoplastic agents
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- the present invention relates to the synthesis of specific N-acridine thiazolidine derivatives and their respective uses as drugs used in cancer therapy.
- Acridine derivatives are known to have a broad spectrum of biological activities, such as antimicrobial, antimalarial, antitripanosomal activities (BONSE et al., 1999, J. Med. Chem., V.42, n.26, p.5448-54) , leishmanicide (GIRAULT et al., 2000, J. Med. Chem., v.43, n.14, p.2646-54) and, above all, for their antineoplastic properties.
- the medicinal interest of acridines dates from 1888, but only in 1913 did these compounds begin to be used in medical practice when Browning discovered the bactericidal action of proflavin c and acriflavin b.
- Anticancer activity was first considered in 1920. Thereafter, various compounds, natural alkaloids or synthetic molecules, were tested as antitumor agents (DEMEUNYNCK et al., 2001, Curr. Pharm. Des., V.7, p. 1703-24).
- the cytotoxic properties of acridine compounds depend on their ability to intercalate between DNA base pairs and also to inhibit nucleic acid synthesis by blocking the action of the topoisomerase I and topoisomerase II enzymes (SURDON et al., 2001, Molecules, v.6, p.673-82). Therefore, these compounds exert their primary clinical effects by their ability to interfere with DNA function, either by inhibiting DNA replication or transcription.
- thiazacridine and imidazacridine derivatives that proved to be very effective in treating cancer.
- the synthetic route used by the same authors to obtain thiazacridine derivatives began with the oxidation of 9-methyl acridine prepared from diphenylamine to acridine-9-carboxaldehyde followed by a Knoevenagel-type condensation reaction. alkaline with ethyl cyanoacetate to obtain ethyl 2-cyano-acridin-9-yl-acrylate ester.
- thiazolidine part of the molecule was performed by benzylation of thiazolidine-2,4-dione with benzyl halides in alkaline medium.
- thiazacridine derivatives were obtained by an addition reaction with ethyl 2-cyano-acridin-9-yl-acrylate ester in the presence of piperidine (PITTA et al, 2002, PI 0203747-5, 2003, PCT / BR03 / 00128; PITTA et al., 2006, PI-0601827-0, PCT / WO 2007/109871 A2.
- the substances obtained according to this invention comprise a group of N-acridinic thiazolidine derivatives.
- Ethyl 2-cyano-acridin-9-yl-acrylate derivative (Figure 1) was used in condensation with 3-acridin-9-ylmethyl-thiazolidine-2,4-dione LPSF AA-1A ( Figure 2) to obtain bis-acridinium compound 3-acridin-9-ylmethyl-5-acridin-9-ylmethylene-thiazolidine-2,4-dione LPSF AA-2 ( Figure 3).
- the ⁇ / s-acridine derivative is obtained by heating 3- (acridin-9-yl-methyl) -thiazolidine-2,4-dione, dissolved in anhydrous ethanol in the presence of piperidine, with 2-cyano-acridin. Ethyl 9-yl-acrylate at a temperature of 80 ° C for 4 hours.
- HL60 leukemia
- MDA-MB 435 breast - human
- HCT-8 human - colon
- RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% antibiotics, kept in an oven at 37 ° C and an atmosphere containing 5% CO 2 .
- Cells were plated at a concentration of 0.3 x 10 6 cells / 100 ⁇ L for suspended cells and 0.1 x 10 6 ⁇ 8 / 100 ⁇ for adhered cells.
- the various compounds were added at different concentrations, ranging from 0.39 - 25ug / mL for pure substances. They were incubated for 72 hours in a 5% CO 2 oven at 37 ° C. At the end of this, the plates were centrifuged and the supernatant removed. Then 200 ⁇ of MTT solution (tetrazolium salt) was added, and the plates were incubated for 3h. The absorbance was read after dissolution of the precipitate with DMSO in a 550nm plate photometer spectrum.
- MTT solution tetrazolium salt
- the compounds were diluted to a concentration of 5mg / mL. 100 ⁇ g / mL were tested in single concentration on the following SF295 (CNS) strains; HCT-8 (colon) and MDA-MB-435 (breast). The compounds were selected according to tumor growth inhibition percentage higher than 80% in the cell lines used (GI%> 80%). These compounds were tested for IC50 determination.
- cytotoxic activity was presented in table 1, which refers to proliferation inhibition (%) performed in duplicate by the MTT method for SF-295 (CNS), HCT-8 (colon carcinoma) and MDA-MB435 (melanoma) cells; doxorubicin was used as a positive control; Table 1 shows that values of 0 - 35% were considered (SA) - No Activity; 36 - 55% (PA) - Little Activity; 56 - 85% (MO) - Moderate Activity; 86 - 100% (MA) - Lots of Activity.
- the LPSF-AA2 derivative was the most active molecule in this series. All compounds show a selective cytotoxic potential for the HCT-8 cell line, with compound AA6 showing the highest cytotoxic potential for this cell line.
- the LPSF-AA6 compound with a bromine atom for substituted on the benzylidene ring, was also quite effective. It was the most active for the HCT-8 cell line and the second most active for the SF-295 and MDA-MB435 cell lines.
- LPSF-AA5 was the least active of all compounds synthesized against cell line HCT-8 and MDA-MB435.
- LPSF-AA2, LPSF-AA3 and LPSF-AA6 were selected for the determination of IC 5 o (Table 2), since they presented tumor growth inhibition percentage higher than 80% o in the used cell lines.
- IC50 values (50% inhibitory concentration) and 95% confidence interval (95% CI) performed by the MTT method for cells HL-60 (promyelocytic leukemia), CEM (lymphocytic leukemia), MDA-MB435 (melanoma), HCT-8 (colon carcinoma) and SF-295 (CNS) obtained by nonlinear regression using the GraphPad Prism program
- LPSF-AA2 had the best IC50 values: 4.4 ⁇ g / mL for MDA-MB435, 4.45 ⁇ g / mL for HCT-8, 7.01 ⁇ g / mL for SF-295 and values greater than 25 ⁇ g / mL for HL-60 and CEM, but still these values are very high when compared to doxorubicone.
- Figure 1 Representation of ethyl 2-cyano-acridin-9-yl-acrylate ester.
- Figure 2 Representation of the intermediate acridine molecule: 3-acridin-9-ylmethyl-thiazolidine-2,4-dione - LPSF AA-1 A.
- Figure 3 Representation of the ⁇ zs-acridine derivative 3-acridm-9-ylmethyl-5-acridin-9-ylmethylene-thiazolidine-2,4-dione - LPSF AA-2.
- Figure 4 - presents the general structure of seven of the nine synthesized derivatives.
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Abstract
Description
TIAZACRIDINAS UTILIZADAS NA TERAPIA ANTICÂNCER TIAZACRIDINES USED IN ANTICANUS THERAPY
OBJETO DA INVENÇÃO OBJECT OF THE INVENTION
A presente invenção refere-se à síntese de derivados tiazolidínicos N-acridínicos específicos e seus respectivos usos como fármacos utilizados na terapêutica do câncer. The present invention relates to the synthesis of specific N-acridine thiazolidine derivatives and their respective uses as drugs used in cancer therapy.
ESTADO DA TÉCNICA TECHNICAL STATE
Os derivados acridínicos são conhecidos por apresentarem um amplo espectro de atividades biológicas, como atividades antimicrobiana, antimalárica, antitripanossômica (BONSE et al., 1999, J. Med. Chem., v.42, n.26, p.5448-54), leishmanicida (GIRAULT et al., 2000, J. Med. Chem., v.43, n.14, p.2646-54) e, sobretudo, por suas propriedades antineoplásicas. O interesse medicinal das acridinas data de 1888, mas apenas em 1913 estes compostos começaram a ser usados na prática médica quando Browning descobriu a ação bactericida da proflavina c e da acriflavina b. A atividade anticâncer foi considerada pela primeira vez em 1920. A partir de então, vários compostos, alcalóides naturais ou moléculas sintéticas, foram testados como agentes antitumorais (DEMEUNYNCK et al., 2001, Curr. Pharm. Des., v.7, p.1703-24). As propriedades citotóxicas dos compostos acridínicos dependem de sua habilidade de se intercalar entre os pares de base do DNA e, também, de inibir a síntese de ácidos nucléicos pelo bloqueio da ação das enzimas topoisomerase I e topoisomerase II (SURDON et al., 2001 , Molecules, v.6, p.673-82). Logo, estes compostos exercem seus efeitos clínicos primários pela capacidade de interferir na função do DNA, seja inibindo a replicação ou transcrição do DNA. Pesquisadores do Laboratório de Planejamento e Síntese de Fármacos - LPSF realizaram a síntese de derivados tiazacridínicos e imidazacridínicos que mostraram ser bastante eficazes no tratamento do câncer. A rota sintética utilizada pelos mesmos autores para a obtenção dos derivados tiazacridínicos iniciou-se com a oxidação da 9-metil-acridina, preparada a partir da difenilamina, em acridina-9-carboxaldeído, seguindo-se uma reação de condensação tipo Knoevenagel em meio alcalino com o cianoacetato de etila para obtenção do éster 2-ciano-acridina-9-il-acrilato de etila. A síntese da parte tiazolidínica da molécula foi realizada por meio da benzilação da tiazolidina-2,4-diona com os halogenetos de benzila em meio alcalino. Na última etapa, os derivados tiazacridídinos foram obtidos através de uma reação de adição com o éster 2-ciano-acridina-9-il-acrilato de etila na presença de piperidina (PITTA et al, 2002, PI 0203747-5, 2003, PCT/BR03/00128; PITTA et al., 2006, PI-0601827- 0, PCT/WO 2007/109871 A2. Acridine derivatives are known to have a broad spectrum of biological activities, such as antimicrobial, antimalarial, antitripanosomal activities (BONSE et al., 1999, J. Med. Chem., V.42, n.26, p.5448-54) , leishmanicide (GIRAULT et al., 2000, J. Med. Chem., v.43, n.14, p.2646-54) and, above all, for their antineoplastic properties. The medicinal interest of acridines dates from 1888, but only in 1913 did these compounds begin to be used in medical practice when Browning discovered the bactericidal action of proflavin c and acriflavin b. Anticancer activity was first considered in 1920. Thereafter, various compounds, natural alkaloids or synthetic molecules, were tested as antitumor agents (DEMEUNYNCK et al., 2001, Curr. Pharm. Des., V.7, p. 1703-24). The cytotoxic properties of acridine compounds depend on their ability to intercalate between DNA base pairs and also to inhibit nucleic acid synthesis by blocking the action of the topoisomerase I and topoisomerase II enzymes (SURDON et al., 2001, Molecules, v.6, p.673-82). Therefore, these compounds exert their primary clinical effects by their ability to interfere with DNA function, either by inhibiting DNA replication or transcription. Researchers at the Laboratory of Drug Planning and Synthesis - LPSF performed the synthesis of thiazacridine and imidazacridine derivatives that proved to be very effective in treating cancer. The synthetic route used by the same authors to obtain thiazacridine derivatives began with the oxidation of 9-methyl acridine prepared from diphenylamine to acridine-9-carboxaldehyde followed by a Knoevenagel-type condensation reaction. alkaline with ethyl cyanoacetate to obtain ethyl 2-cyano-acridin-9-yl-acrylate ester. The synthesis of the thiazolidine part of the molecule was performed by benzylation of thiazolidine-2,4-dione with benzyl halides in alkaline medium. In the last step, thiazacridine derivatives were obtained by an addition reaction with ethyl 2-cyano-acridin-9-yl-acrylate ester in the presence of piperidine (PITTA et al, 2002, PI 0203747-5, 2003, PCT / BR03 / 00128; PITTA et al., 2006, PI-0601827-0, PCT / WO 2007/109871 A2.
DESCRIÇÃO DA INVENÇÃO DESCRIPTION OF THE INVENTION
As substâncias obtidas conforme esta invenção compõem um grupo de derivados tiazolidínicos N-acridinicos. O derivado 2-ciano-acridina-9-il-acrilato de etila (Figura 1) foi utilizado na condensação com o 3-acridin-9-ilmetil-tiazolidina-2,4-diona LPSF AA-1A (Figura 2) para obtenção do composto bis-acridinico 3-acridin-9-ilmetil-5-acridin-9- ilmetileno-tiazolidina-2,4-diona LPSF AA-2 (Figura 3). Os derivados de formula geral 3- acridin-9-ilmetil-5-benzilideno-tiazolidina-2,4-diona (Figura 4) foram obtidos a partir da reação tipo Michael dos derivados 2-ciano-3-fenil-acrilatos de etila com o derivado 3-acridin- 9-ilmetil-tiazolidina-2,4-diona LPSF AA-1A (Figura 2). Os ésteres 2-ciano-3-fenil-acrilatos de etila substituídos no anel aromático por metoxi, metil, cloro, bromo ou metanosulfonil foram preparados através da condensação de Knoevenagel a partir dos benzaldeidos correspondentes com o cianacetato de etila. The substances obtained according to this invention comprise a group of N-acridinic thiazolidine derivatives. Ethyl 2-cyano-acridin-9-yl-acrylate derivative (Figure 1) was used in condensation with 3-acridin-9-ylmethyl-thiazolidine-2,4-dione LPSF AA-1A (Figure 2) to obtain bis-acridinium compound 3-acridin-9-ylmethyl-5-acridin-9-ylmethylene-thiazolidine-2,4-dione LPSF AA-2 (Figure 3). Derivatives of formula 3-acridin-9-ylmethyl-5-benzylidene-thiazolidine-2,4-dione (Figure 4) were obtained from Michael-type reaction of ethyl 2-cyano-3-phenyl-acrylates derivatives with 3-acridin-9-ylmethyl-thiazolidine-2,4-dione derivative LPSF AA-1A (Figure 2). Ethyl 2-cyano-3-phenyl-acrylates substituted on the aromatic ring by methoxy, methyl, chlorine, bromine or methanesulfonyl were prepared by condensing Knoevenagel from the corresponding benzaldehydes with ethyl cyanacetate.
A. Síntese dos Compostos A. Synthesis of Compounds
Procedimento geral de preparação dos Ésteres 2-ciano-3-fenil-acrilatos de etila substituídos General Procedure for Preparation of Substituted Ethyl 2-Cyano-3-Phenyl Acrylates Esters
Em um balão de fundo redondo, com agitação magnética, adaptado a um condensador, foram adicionados quantidades èquimolares do aldeído aromático substituído e do cianoacetato de etila na presença de piperidina como catalisador e benzeno como solvente. A mistura reacional foi aquecida sob refluxo a 110°C por 4 horas. Passado este tempo, o produto foi seco, suspenso em água, filtrado e purificado com álcool etílico. In a magnetically stirred, round-bottomed flask fitted with a condenser, ions were added to the chemimolar amounts of the substituted aromatic aldehyde and ethyl cyanoacetate in the presence of piperidine as catalyst and benzene as solvent. The reaction mixture was heated at reflux at 110 ° C for 4 hours. After this time, the product was dried, suspended in water, filtered and purified with ethyl alcohol.
Procedimento de preparação do éster 2-ciano-acridina-9-il-acrilato de etila (Figura 1) Em um balão de undo redondo, com agitação magnética, adaptado a um condensador, foram adicionadas quantidades èquimolares da a 9-acridina-carboxaldeído e do cianoacetato de etila na presença de piperidina como catalizador. A mistura reacional foi aquecida sob refluxo a 110°C por 8 horas. O produto foi purificado por cromatografia sob pressão em sílica gel 60, rc-hexano/acetato 6:4. Procedimento de preparação da 3-acridin-9-ilmetil-tiazolidina-2,4-diona codificado LPSF AA-1A (Figura 2) Ethyl 2-cyano-acridin-9-yl-acrylate ester preparation procedure (Figure 1) In a round, magnetic stirring flask fitted to a condenser, the β-acridine carboxaldehyde and ε-molecular amounts were added. of cyanoacetate in the presence of piperidine as catalyst. The reaction mixture was heated at reflux at 110 ° C for 8 hours. The product was purified by chromatography on silica gel 60, 6-hexane / acetate 6: 4. Preparation procedure for LPSF AA-1A encoded 3-acridin-9-ylmethyl-thiazolidine-2,4-dione (Figure 2)
Em um balão de fundo redondo, com agitação magnética, adaptado a um condensador, foram adicionados a tiazolidina-2,4-diona e hidróxido de sódio previamente dissolvido em álcool etílico. A mistura reacional foi agitada durante 10 minutos. Passado este tempo, foi adicionado a 9-bromometilacridina em quantidades èquimolares e a mistura foi aquecida a uma temperatura de 60°C durante 7 horas. O produto foi lavado com água destilada e, depois, purificado com álcool etílico. In a magnetic stirring, round-bottomed flask fitted with a condenser were added thiazolidine-2,4-dione and sodium hydroxide previously dissolved in ethyl alcohol. The reaction mixture was stirred for 10 minutes. After this time 9-bromomethylacridine was added in equimolar amounts and the mixture was heated at a temperature of 60 ° C for 7 hours. The product was washed with distilled water and then purified with ethyl alcohol.
Procedimento de preparação da 3-acridin-9-ilmetil-5-acridin-9-iImetileno-tiazolidina- 2,4-diona LPSF AA-2 (Figura 3) 3-Acridin-9-ylmethyl-5-acridin-9-ylmethylene-thiazolidine-2,4-dione LPSF AA-2 preparation procedure (Figure 3)
Obtêm-se o derivado è/s-acridínico através do aquecimento da 3-(acridin-9-il-metil)- tiazolidina-2,4-diona , dissolvida em etanol anidro na presença de piperidina, com o 2-ciano- acridin-9-il-acrilato de etila a uma temperatura de 80°C por 4 horas. The ε / s-acridine derivative is obtained by heating 3- (acridin-9-yl-methyl) -thiazolidine-2,4-dione, dissolved in anhydrous ethanol in the presence of piperidine, with 2-cyano-acridin. Ethyl 9-yl-acrylate at a temperature of 80 ° C for 4 hours.
Procedimento geral de preparação das tiazacridinas substituídas (Figura 4) General procedure for preparing substituted thiazacridines (Figure 4)
Em um balão de fundo redondo, com agitação magnética, adaptado a um condensador, foram adicionados quantidades èquimolares da 3-acridin-9-ilmetil-tiazolidina-2,4-diona e dos derivados 2-ciano-3-fenil-acrilatos de etila substituídos dissolvidos em álcool etílico, na presença de piperidina como catalisador. A mistura reacional foi aquecida sob refluxo a 50°C por 4 horas levando à formação dos novos derivados tiazacrinícos, que foram purificados através de lavagens sucessivas com água destilada e álcool etílico. Foram sintetizados os compostos de formula geral 3-acridin-9-ilmetil-5-benzilideno-tiazolidina-2,4-diona (Figura 4): In a magnetic stirring, round-bottomed flask fitted with a condenser, the amount of 3-acridin-9-ylmethyl-thiazolidine-2,4-dione and ethyl 2-cyano-3-phenyl-acrylates derivatives were added. Substitutes dissolved in ethyl alcohol in the presence of piperidine as catalyst. The reaction mixture was heated to reflux at 50 ° C for 4 hours leading to the formation of new thiazacrinic derivatives, which were purified by successive washes with distilled water and ethyl alcohol. The compounds of formula 3-acridin-9-ylmethyl-5-benzylidene-thiazolidine-2,4-dione were synthesized (Figure 4):
3-Acridin-9-ilmetil-5-(4-metoxi-benzilideno)-tiazolidina-2,4-diona (LPSF AA-3) 3-Acridin-9-ylmethyl-5- (4-methoxy-benzylidene) -thiazolidine-2,4-dione (LPSF AA-3)
3-Acridin-9-ilmetil-5-(4-metil-benzilideno)-tiazolidina-2,4-diona (LPSF AA-4) 3-Acridin-9-ylmethyl-5- (4-methyl-benzylidene) -thiazolidine-2,4-dione (LPSF AA-4)
3-Acridin-9-ilmetil-5-(4-cloro-benzilideno)-tiazolidina-2,4-diona (LPSF AA-5) 3-Acridin-9-ylmethyl-5- (4-chloro-benzylidene) -thiazolidine-2,4-dione (LPSF AA-5)
3-Acridin-9-ilmetil-5-(4-bromo-benzilideno)-tiazolidina-2,4-diona (LPSF AA-6) 3-Acridin-9-ylmethyl-5- (4-bromo-benzylidene) -thiazolidine-2,4-dione (LPSF AA-6)
3-Acridin-9-ilmetil-5-(4-metanosulfonil-benzilideno)-tiazolidina-2,4-diona (LPSF AA-8) 3-Acridin-9-ilmetil-5-(3-bromo-4-metoxi-benzilideno)-tiazolidina-2,4-diona (LPSF AA-9) B. Avaliação da Atividade Anticâncer Análise de citotoxicidade pelo método do MTT 3-Acridin-9-ylmethyl-5- (4-methanesulfonyl-benzylidene) -thiazolidin-2,4-dione (LPSF AA-8) 3-Acridin-9-ylmethyl-5- (3-bromo-4-methoxy) benzylidene) thiazolidine-2,4-dione (LPSF AA-9) B. Evaluation of Anticancer Activity Cytotoxicity analysis by the MTT method
Células: As linhagens utilizadas, HL60 (leucemia), MDA-MB 435(mama - humano), HCT-8 (cólon - humano), foram cedidas pelo Instituto Nacional do Câncer (USA), tendo sido cultivadas em meio RPMI 1640, suplementados com 10% de soro fetal bovino e 1% de antibióticos, mantidas em estufa a 37°C e atmosfera contendo 5% de C02. Cells: The strains used, HL60 (leukemia), MDA-MB 435 (breast - human), HCT-8 (human - colon), were provided by the National Cancer Institute (USA) and were grown in RPMI 1640 medium supplemented with with 10% fetal bovine serum and 1% antibiotics, kept in an oven at 37 ° C and an atmosphere containing 5% CO 2 .
As células foram plaqueadas na concentração de 0,3 x IO6 céls./100μL, para células suspensas e 0,1 x IO6 ϋέΐ8./100μί, para células aderidas. Os diversos compostos foram acrescidos em diferentes concentrações, que variaram entre 0,39 - 25ug/mL para substâncias puras. Foram incubadas por 72 horas em estufa a 5% de C02 a 37°C. Ao término deste, as placas foram centrifugadas e o sobrenadante foi removido. Em seguida, foram adicionados 200μί da solução de MTT (sal de tetrazolium), e as placas foram incubadas por 3h. A absorbância foi lida após dissolução do precipitado com DMSO em espectro fotômetro de placa a 550nm.Cells were plated at a concentration of 0.3 x 10 6 cells / 100μL for suspended cells and 0.1 x 10 6 ϋέΐ8 / 100μί for adhered cells. The various compounds were added at different concentrations, ranging from 0.39 - 25ug / mL for pure substances. They were incubated for 72 hours in a 5% CO 2 oven at 37 ° C. At the end of this, the plates were centrifuged and the supernatant removed. Then 200μί of MTT solution (tetrazolium salt) was added, and the plates were incubated for 3h. The absorbance was read after dissolution of the precipitate with DMSO in a 550nm plate photometer spectrum.
Os experimentos foram analisados segundo suas médias e respectivos erros-padrão. O cálculo das IC5o (concentração inibitória média capaz de provocar 50% do efeito máximo) e seus respectivos desvios foram realizados a partir da regressão não-linear no programa GraphPad Prism. Cada amostra foi analisada a partir de 2 dois experimentos realizados em triplicata.The experiments were analyzed according to their means and standard errors. The calculation of IC 50 (mean inhibitory concentration capable of causing 50% of the maximum effect) and their respective deviations were performed from nonlinear regression in the GraphPad Prism program. Each sample was analyzed from 2 two experiments performed in triplicate.
Os compostos foram diluídos na concentração de 5mg/mL. Foram testados em concentração única 100 μg/mL nas seguintes linhagens SF295 (SNC); HCT-8 (cólon) e MDA-MB-435 (mama). Os compostos foram selecionados de acordo com percentual de inibição do crescimento tumoral maior que 80% nas linhagens celulares utilizadas (GI% > 80%). Estes compostos foram testados para a determinação do IC50. The compounds were diluted to a concentration of 5mg / mL. 100 µg / mL were tested in single concentration on the following SF295 (CNS) strains; HCT-8 (colon) and MDA-MB-435 (breast). The compounds were selected according to tumor growth inhibition percentage higher than 80% in the cell lines used (GI%> 80%). These compounds were tested for IC50 determination.
Resultados Results
Os derivados tiazacridínicos foram testados nas seguintes linhagens: SF-295, HCT-8 e MDA- MB435. Os resultados da atividade citotóxica estão apresentados na tabela 1 , que se refere a inibição da proliferação (%) realizado em duplicata pelo método do MTT para as células SF- 295 (SNC), HCT-8 (carcinoma de cólon) e MDA-MB435 (melanoma); doxorrubicina foi usada como controle positivo, na tabela 1 temos que os valores de 0 - 35% foram considerados (SA) - Sem Atividade; 36 - 55% (PA) - Pouca Atividade; 56 - 85% (MO) - Moderada Atividade; 86 - 100% (MA) - Muita Atividade. Thiazacridine derivatives were tested in the following strains: SF-295, HCT-8 and MDA-MB435. The results of cytotoxic activity are presented in table 1, which refers to proliferation inhibition (%) performed in duplicate by the MTT method for SF-295 (CNS), HCT-8 (colon carcinoma) and MDA-MB435 (melanoma) cells; doxorubicin was used as a positive control; Table 1 shows that values of 0 - 35% were considered (SA) - No Activity; 36 - 55% (PA) - Little Activity; 56 - 85% (MO) - Moderate Activity; 86 - 100% (MA) - Lots of Activity.
Tabela 1 Table 1
Os compostos LPSF-AA2, LPSF-AA3 e LPSF-AA6 foram selecionados para a determinação do IC5o (Tabela 2), pois apresentaram percentual de inibição do crescimento tumoral maior que 80%o nas linhagens celulares utilizadas. Valores de IC50 (concentração inibitória de 50%) e intervalo de confiança de 95% (IC 95%) realizado pelo método do MTT para as células HL- 60 (leucemia promielocítica), CEM (leucemia linfocítica), MDA-MB435 (melanoma), HCT-8 (carcinoma de cólon) e SF-295 (SNC) obtidos por regressão não-linear através do programa GraphPad Prism The compounds LPSF-AA2, LPSF-AA3 and LPSF-AA6 were selected for the determination of IC 5 o (Table 2), since they presented tumor growth inhibition percentage higher than 80% o in the used cell lines. IC50 values (50% inhibitory concentration) and 95% confidence interval (95% CI) performed by the MTT method for cells HL-60 (promyelocytic leukemia), CEM (lymphocytic leukemia), MDA-MB435 (melanoma), HCT-8 (colon carcinoma) and SF-295 (CNS) obtained by nonlinear regression using the GraphPad Prism program
Tabela 2 - Table 2 -
Dos 3 compostos mais potentes, o LPSF-AA2 apresentou os melhores valores de IC50: 4,4 μg/mL para MDA-MB435, 4,45 μg/mL para HCT-8, 7,01 μg/mL para SF-295 e valores maiores que 25 μg/mL para HL-60 e CEM, mas ainda assim estes valores estão muito altos quando comparados a doxorrubicona. Of the 3 most potent compounds, LPSF-AA2 had the best IC50 values: 4.4 μg / mL for MDA-MB435, 4.45 μg / mL for HCT-8, 7.01 μg / mL for SF-295 and values greater than 25 μg / mL for HL-60 and CEM, but still these values are very high when compared to doxorubicone.
RELAÇÃO DAS FIGURAS List of Figures
Figura 1 - Representação do éster 2-ciano-acridina-9-il-acrilato de etila. Figure 1 - Representation of ethyl 2-cyano-acridin-9-yl-acrylate ester.
Figura 2 - Representação da molécula acridínica intermediária: 3-acridin-9-ilmetil-tiazolidina- 2,4-diona - LPSF AA-1 A. Figure 2 - Representation of the intermediate acridine molecule: 3-acridin-9-ylmethyl-thiazolidine-2,4-dione - LPSF AA-1 A.
Figura 3 - Representação do derivado òzs-acridínico 3-acridm-9-ilmetil-5-acridin-9- ilmetileno-tiazolidina-2,4-diona - LPSF AA-2. Figure 3 - Representation of the αzs-acridine derivative 3-acridm-9-ylmethyl-5-acridin-9-ylmethylene-thiazolidine-2,4-dione - LPSF AA-2.
Figura 4 - apresenta a estrutura geral de sete dos nove derivados sintetizados. Figure 4 - presents the general structure of seven of the nine synthesized derivatives.
RELAÇÃO DAS TABELAS TABLE LIST
Tabela 1. Valores referentes à determinação da atividade cito tóxica Table 1. Values for determination of cytotoxic activity
Tabela 2. Valores referentes à determinação de IC50 para os compostos LPSF-AA2, LPSF- AA3 e LPSF-AA6 Table 2. IC50 Determination Values for LPSF-AA2, LPSF-AA3, and LPSF-AA6
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