RESUMO
Chlorambucil is an alkylating drug that finds application towards chemotherapy of different types of cancers. In order to explore the possibility of utilization of this drug as an imaging agent for early diagnosis of solid tumors, attempt was made to synthesize a 99mTc complex of chlorambucil and evaluate its potential in tumor bearing small animal model. HYNIC-chlorambucil was synthesized by conjugation of HYNIC with chlorambucil via an ethylenediamine linker. All the intermediates and final product were purified and characterized by standard spectroscopic techniques viz. FT-IR, 1H/13C-NMR as well as by mass spectrometry. HYNIC-chlorambucil conjugate was radiolabeled with [99mTc]Tc and found to be formed with > 95 % radiochemical purity via RP-HPLC studies. The partition coefficient (Log10Po/w) of the synthesized complex was found to be -0.78 ± 0.25 which indicated the moderate hydrophilic nature for the complex. Biological behaviour of [99mTc]Tc-HYNIC-chlorambucil, studied in fibrosarcoma bearing Swiss mice, revealed a tumor uptake of about 4.16 ± 1.52 %IA/g at 30 min post-administration, which declined to 1.91 ± 0.13 % IA/g and 1.42 ± 0.14 %IA/g at 1 h and 2 h post-administration, respectively. A comparison of different [99mTc]Tc-chlorambucil derivatives (reported in the contemporary literature) formulated using different methodologies revealed that tumor uptake and pharmacokinetics exhibited by these agents strongly depend on the lipophilicity/hydrophilicity of such agents, which in turn is dependent on the bifunctional chelators used for formulating the radiolabeled chlorambucils.
Assuntos
Clorambucila , Compostos de Organotecnécio , Animais , Humanos , Camundongos , Antineoplásicos Alquilantes/síntese química , Antineoplásicos Alquilantes/química , Antineoplásicos Alquilantes/farmacologia , Linhagem Celular Tumoral , Clorambucila/química , Clorambucila/síntese química , Clorambucila/farmacologia , Estrutura Molecular , Ácidos Nicotínicos/química , Ácidos Nicotínicos/síntese química , Compostos de Organotecnécio/química , Compostos de Organotecnécio/síntese química , Compostos de Organotecnécio/farmacocinética , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/química , Tecnécio/química , Distribuição TecidualRESUMO
A pH-responsive smart nanocarrier with significant components was synthesized by conjugating the non-emissive anticancer drug methyl orange and polyethylene glycol derived folate moiety to the backbone of polynorbornene. Complete synthesis procedure and characterization methods of three monomers included in the work: norbornene-derived Chlorambucil (Monomerâ 1), norbornene grafted with polyethylene glycol, and folic acid (Monomerâ 2) and norbornene attached methyl orange (Monomerâ 3) connected to the norbornene backbone through ester linkage were clearly discussed. Finally, the random copolymer CHO PEG FOL METH was synthesized by ring-opening metathesis polymerization (ROMP) using Grubbs' second-generation catalyst. Advanced polymer chromatography (APC) was used to find the final polymer's molecular weight and polydispersity index (PDI). Dynamic light scattering, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were utilized to explore the prodrug's size and morphology. Release experiments of the anticancer drug, Chlorambucil and the coloring agent, methyl orange, were performed at different pH and time. Cell viability assay was carried out for determining the rate of survived cells, followed by the treatment of our final polymer named CHO PEG FOL METH.
Assuntos
Antineoplásicos/química , Portadores de Fármacos/química , Ácido Fólico/análogos & derivados , Plásticos/química , Polietilenoglicóis/química , Pró-Fármacos/química , Antineoplásicos/síntese química , Antineoplásicos/toxicidade , Compostos Azo/síntese química , Compostos Azo/química , Compostos Azo/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Clorambucila/toxicidade , Corantes/síntese química , Corantes/química , Corantes/toxicidade , Preparações de Ação Retardada/síntese química , Preparações de Ação Retardada/química , Preparações de Ação Retardada/toxicidade , Doxorrubicina/síntese química , Doxorrubicina/química , Doxorrubicina/toxicidade , Portadores de Fármacos/síntese química , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Ácido Fólico/síntese química , Ácido Fólico/química , Ácido Fólico/toxicidade , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Plásticos/síntese química , Plásticos/toxicidade , Polietilenoglicóis/síntese química , Polietilenoglicóis/toxicidade , Polimerização , Pró-Fármacos/síntese química , Pró-Fármacos/toxicidadeRESUMO
Chlorambucil is a nitrogen mustard-based DNA alkylating drug, which is widely used as a front-line treatment of chronic lymphocytic leukaemia (CLL). Despite its widespread application and success for the initial treatment of leukaemia, a majority of patients eventually develop acquired resistance to chlorambucil. In this regard, we have designed and synthesised a novel hybrid molecule, chloram-HDi that simultaneously impairs DNA and HDAC enzymes. Chloram-HDi efficiently inhibits the proliferation of HL-60 and U937 leukaemia cells with GI50 values of 1.24 µM and 1.75 µM, whereas chlorambucil exhibits GI50 values of 21.1 µM and 37.7 µM against HL-60 and U937 leukaemia cells, respectively. The mechanism behind its remarkably enhanced cytotoxicity is that chloram-HDi not only causes a significant DNA damage of leukaemia cells but also downregulates DNA repair protein, Rad52, resulting in the escalation of its DNA-damaging effect. Furthermore, chloram-HDi inhibits HDAC enzymes to induce the acetylation of α-tubulin and histone H3.
Assuntos
Antineoplásicos/farmacologia , Clorambucila/farmacologia , DNA de Neoplasias/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/metabolismo , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/química , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Dano ao DNA , DNA de Neoplasias/química , Relação Dose-Resposta a Droga , Desenho de Fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores de Histona Desacetilases/síntese química , Inibidores de Histona Desacetilases/química , Humanos , Leucemia Linfocítica Crônica de Células B/metabolismo , Leucemia Linfocítica Crônica de Células B/patologia , Estrutura Molecular , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
l-type amino acid transporter 1 (LAT1) is an amino acid transporter that is overexpressed in several types of cancer and, thus, it can be a potential target for chemotherapy. The objectives of this study were to (a) synthesize LAT1-targeted chlorambucil derivatives and (b) evaluate their LAT1-mediated cellular uptake as well as antiproliferative activity in vitro in the human breast cancer MCF-7 cell line. Chlorambucil was conjugated to l-tyrosine-an endogenous LAT1 substrate-via either ester or amide linkage (compounds 1 and 2, respectively). While chlorambucil itself did not bind to LAT1, its derivatives 1 and 2 bound to LAT1 with a similar affinity as with l-tyrosine and their respective cellular uptake was significantly higher than that of chlorambucil in MCF-7. The results of our cellular uptake study are indicative of antiproliferative activity, as a higher intracellular uptake of chlorambucil derivatives resulted in greater cytotoxicity than chlorambucil by itself. LAT1 thus contributes to intracellular uptake of chlorambucil derivatives and, therefore, increases antiproliferative activity. The understanding gained from our research can be used in the development of LAT1-targeted anticancer drugs and prodrugs for site-selective and enhanced chemotherapeutic activity.
Assuntos
Neoplasias da Mama/metabolismo , Clorambucila/farmacologia , Endocitose , Transportador 1 de Aminoácidos Neutros Grandes/metabolismo , Tirosina/farmacologia , Neoplasias da Mama/patologia , Proliferação de Células/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Endocitose/efeitos dos fármacos , Feminino , Humanos , Células MCF-7 , Ligação Proteica/efeitos dos fármacos , Fatores de Tempo , Tirosina/químicaRESUMO
BACKGROUND: One of the possible ways of improving the activity and selectivity profile of anticancer agents is to design drug carrier systems employing nanomolecules. Calix[4]arene derivatives and chlorambucil and ibuprofen are important compounds that exhibit interesting anticancer properties. OBJECTIVE: The objective of this article is the synthesis of new calix[4]arene-derivative conjugates of chlorambucil or ibuprofen with potential anticancer activity. METHODS: Cytotoxicity assays were determined using the protein-binding dye sulforhodamine B (SRB) in microculture to measure cell growth as described [19, 20]. Conjugates of chlorambucil and resorcinarene-dendrimers were prepared in 2% DMSO and added into the culture medium immediately before use. Control cells were treated with 2% DMSO. RESULTS: Thus, calix[4]arene-derivative conjugates of chlorambucil or ibuprofen showed good stability of the chemical link between drug and spacer. Evaluation of the cytotoxicity of the calix[4]arene chlorambucil or ibuprofen conjugates employing a sulforhodamine B (SRB) assay in K-562 (human chronic myelogenous leukemia cells) and U-251 (human glioblastoma cells) demonstrated that the conjugate was more potent as an antiproliferative agent than free chlorambucil and ibuprofen. The conjugates did not show any activity against the COS-7 African green monkey kidney fibroblast cell line. CONCLUSION: In the paper, we report the synthesis and spectroscopic analyses of new calix[4]arene derivative conjugates of chlorambucil or ibuprofen. Cytotoxicity assays revealed that at 10 µM, the conjugates were very active against K-562 (human chronic myelogenous leukemia cells) and U- 251 (human glioblastoma cells) cancer cells' proliferation. In order to explain the molecular mechanisms involved in the anticancer activity of calix[4]arene chlorambucil or ibuprofen conjugates, our research will be continued.
Assuntos
Antineoplásicos/farmacologia , Calixarenos/farmacologia , Clorambucila/farmacologia , Ibuprofeno/farmacologia , Fenóis/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Calixarenos/química , Proliferação de Células/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Ibuprofeno/síntese química , Ibuprofeno/química , Estrutura Molecular , Fenóis/química , Células Tumorais CultivadasRESUMO
Here we report the design and synthesis of a chlorambucil-alkynyl (CHL-CCH) ligand, mononuclear gold(i) complex K[(CHL-CC)AuCl], 1, and heteronuclear complex (CHL-CC)Au(µ2-η2-CS3)Ti(η5-Cp)2, 2 for renal cancer. Complex 2 is significantly more cytotoxic than complex 1 and cisplatin against renal cancer cells with a high selectivity index value. The mechanism of action of these complexes against renal cancer cells was studied in detail by experimental and computational methods.
Assuntos
Antineoplásicos/farmacologia , Clorambucila/análogos & derivados , Clorambucila/farmacologia , Complexos de Coordenação/farmacologia , Inibidores Enzimáticos/farmacologia , Tiorredoxina Dissulfeto Redutase/antagonistas & inibidores , Antineoplásicos/síntese química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Clorambucila/síntese química , Complexos de Coordenação/síntese química , Teoria da Densidade Funcional , Desenho de Fármacos , Inibidores Enzimáticos/síntese química , Ouro/química , Humanos , Neoplasias Renais/tratamento farmacológico , Modelos Químicos , Simulação de Acoplamento Molecular , Titânio/químicaRESUMO
BACKGROUND: Scientists have extensively investigated curcumin, yielding many publications on treatments of cancer. Numerous derivatives of curcumin were synthesized, evaluated for their anti-oxidant and free-radical scavenging, SAR, ADME properties and tested in anticancer applications. OBJECTIVE: We decided to exploit curcumin as a bioactive core platform for carrying anticancer drugs, which likely possesses a carboxyl moiety for potential linkage to the carrier for drug delivery. METHODS: The goal of this work is to develop biolabile multifunctional curcumin platforms towards anticancer drug delivery, including determination of drug release profiling in hydrolytic media, in vitro cytotoxicity, antioxidant properties and blockage of relevant cell survival pathways. RESULTS: We report on a facile synthesis of the bioactive multifunctional curcumin-based platforms linked to a variety of anticancer drugs like amonafide and chlorambucil, and release of the drugs in a hydrolytic environment. The leading curcumin-based platform has presented antioxidant activity similar to curcumin, but with much more potent cytotoxicity in vitro in agreement with the augmented blockage of the NF-kB cell survival pathway. CONCLUSION: The approach presented here may prove beneficial for bioactive curcumin-based delivery applications where multiple drug delivery is required in a consecutive and controlled mode.
Assuntos
Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Curcumina/análogos & derivados , Curcumina/farmacologia , Portadores de Fármacos/química , Adenina , Antineoplásicos/síntese química , Antineoplásicos/química , Antioxidantes/síntese química , Antioxidantes/química , Linhagem Celular Tumoral , Clorambucila/síntese química , Clorambucila/química , Clorambucila/farmacologia , Curcumina/síntese química , Portadores de Fármacos/síntese química , Liberação Controlada de Fármacos , Humanos , Naftalimidas/síntese química , Naftalimidas/química , Naftalimidas/farmacologia , Organofosfonatos , Fosforilação/efeitos dos fármacos , Pró-Fármacos/síntese química , Pró-Fármacos/química , Fator de Transcrição RelA/metabolismoRESUMO
Although photothermal therapy (PT) and photothermal-chemotherapy (PT-CT) treatments have been used to achieve complete ablation of solid tumors, they are often implemented at more than 50 °C under high intensity and using a high dose of NIR irradiation, concomitantly inducing heavy skin burning, tissue damage, and ugly scarring. Moreover, the residual tumor cells at the treated site cannot be completely eradicated, resulting in tumor recurrence and metathesis. These key obstacles have prohibited PT and PT-CT treatments from transitioning to clinical use, therefore achieving traceless ablation of solid tumors without recurrence is still a challenge for real applications. To balance hyperthermia and a high drug-loading capacity in polyprodrugs to achieve mild PT-CT, we rationally designed a novel type of intracellular pH and reduction-cleavable chlorambucil prodrug and synthesized high drug-loading polydopamine-chlorambucil conjugate nanoparticles (PDCBs). The PDCBs show good photothermal properties and demonstrate intracellular pH-, reduction-cleavable, and external near infrared (NIR)-triggered drug release profiles. Polydopamine-chlorambucil conjugate nanoparticles with 40 wt% CB (PDCB40) and mild NIR irradiation could facilitate cellular internalization and subcellular trafficking, generating an excellent and synergistic antitumor effect in vitro. Pharmacokinetics and small animal fluorescent and photoacoustic imaging demonstrate that PDCB40 has a 3.6-fold longer blood circulation time compared to free CB and attained selective tumor accumulation, simultaneously inducing a 4.1-fold stronger photoacoustic signal than the control. By using one intravenous injection of PDCB40 and a single dose of mild NIR irradiation, this simple and mild PT-CT treatment achieved a non-discerned tumor on the sixth day, and traceless and complete ablation of a solid MCF-7 tumor without recurrence within 50 days, opening up a new avenue for precise cancer therapy with the potential for real applications.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Nanopartículas/química , Fotoquimioterapia , Pró-Fármacos/farmacologia , Polímeros Responsivos a Estímulos/química , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Neoplasias da Mama/patologia , Proliferação de Células/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Clorambucila/farmacologia , Terapia Combinada , Dopamina/síntese química , Dopamina/química , Dopamina/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Células HeLa , Humanos , Raios Infravermelhos , Células MCF-7 , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Estrutura Molecular , Tamanho da Partícula , Pró-Fármacos/síntese química , Pró-Fármacos/química , Ratos , Ratos Sprague-Dawley , Propriedades de Superfície , Células Tumorais CultivadasRESUMO
The synergistic effect of two anticancer drugs can significantly overcome the multidrug resistance of tumor cells and improve the drug bioavailability. Herein, two different anticancer drugs, camptothecin and chlorambucil, are successfully connected together by a disulfide linkage to get a novel drug-drug conjugated prodrug (G). Using water-soluble pillar[6]arene (WP6) as a host molecule, a supramolecular host-guest complex WP6âG is formed, which can further self-assemble into supramolecular vesicles in aqueous solution. In the specific microenvironment of cancer cells, the disulfide linkage is destroyed and the two anticancer drugs can be released efficiently to achieve a better synergistic effect than a single anticancer drug. Notably, these prodrug nanocarriers can not only effectively kill the cancer cells but also obviously reduce the undesirable side effects on normal cells.
Assuntos
Antineoplásicos/farmacologia , Camptotecina/farmacologia , Clorambucila/farmacologia , Portadores de Fármacos/química , Compostos de Amônio Quaternário/química , Antineoplásicos/síntese química , Antineoplásicos/metabolismo , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Camptotecina/síntese química , Camptotecina/metabolismo , Camptotecina/toxicidade , Clorambucila/síntese química , Clorambucila/metabolismo , Clorambucila/toxicidade , Dissulfetos/síntese química , Dissulfetos/metabolismo , Dissulfetos/farmacologia , Dissulfetos/toxicidade , Portadores de Fármacos/síntese química , Portadores de Fármacos/toxicidade , Glutationa/metabolismo , Humanos , Células MCF-7 , Tamanho da Partícula , Pró-Fármacos/síntese química , Pró-Fármacos/metabolismo , Pró-Fármacos/farmacologia , Pró-Fármacos/toxicidade , Compostos de Amônio Quaternário/síntese química , Compostos de Amônio Quaternário/toxicidade , Solubilidade , Água/químicaRESUMO
Nitrogen mustards, such as chlorambucil (CLB), can cause adverse side-effects due to ubiquitous distribution in non-target organs. To minimize this toxicity, strategies of tumor-targeting drug delivery have been developed, where a cytotoxic warhead is linked to a tumor-cell-specific small ligand. Malignant cells exhibit marked glucose avidity and an accelerated metabolism by aerobic glycolysis, known as the Warburg effect, and recognized as a hallmark of cancer. A targeting approach exploiting the Warburg effect by conjugation of CLB to 2-fluoro-2-deoxyglucose (FDG) was previously reported and identified two peracetylated glucoconjugates 2 and 3 with promising antitumor activities in vivo. These results prompted us to investigate the importance of the spacer in this tumor-targeting glucose-based conjugates. Here we report the chemical synthesis and an in vitro cytotoxicity evaluation, using a 5-member panel of human tumor cell lines and human fibroblasts, of 16 new CLB glucoconjugates in which the alkylating drug is attached to the C-1 position of FDG via different linkages. We studied the structure-activity relationships in the linker, and evidenced the positive impact of an aromatic linker on in vitro cytotoxicity: compound 51 proved to be the most active FDG-CLB glucoside, characterized by a bis-aromatic spacer tethered to CLB through an amide function.
Assuntos
Antígenos de Neoplasias , Clorambucila/química , Sistemas de Liberação de Medicamentos , Fluordesoxiglucose F18/química , Fluordesoxiglucose F18/farmacologia , Antígenos de Neoplasias/química , Antineoplásicos Alquilantes/química , Antineoplásicos Alquilantes/farmacologia , Antineoplásicos Alquilantes/toxicidade , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Clorambucila/síntese química , Clorambucila/farmacologia , Fluordesoxiglucose F18/síntese química , Fluordesoxiglucose F18/toxicidade , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
A FRET donor-acceptor xanthene-coumarin conjugate has been designed for redox-regulated synergic treatment of photodynamic therapy and chemotherapy with real-time monitoring. The "locked" FRET pair was selectively "unlocked" by biological reducing thiols via rupture of a sacrificial disulfide linker. A distinct change in fluorescence color and selective cancer cell toxicity were observed in vitro.
Assuntos
Clorambucila/análogos & derivados , Clorambucila/farmacologia , Pró-Fármacos/farmacologia , Umbeliferonas/farmacologia , Xantenos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Antineoplásicos/efeitos da radiação , Clorambucila/síntese química , Clorambucila/efeitos da radiação , Terapia Combinada/métodos , Tratamento Farmacológico/métodos , Fluorescência , Glutationa/química , Células HEK293 , Células HeLa , Humanos , Luz , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/efeitos da radiação , Pró-Fármacos/síntese química , Pró-Fármacos/efeitos da radiação , Oxigênio Singlete/química , Nanomedicina Teranóstica , Umbeliferonas/síntese química , Umbeliferonas/efeitos da radiação , Xantenos/síntese química , Xantenos/efeitos da radiaçãoRESUMO
Histone deacetylases (HDACs) play a pivotal role not only in gene expression but also in DNA repair. Herein, we report the successful design, synthesis and evaluation of a chlorambucil derivative named vorambucil with a hydroxamic acid tail as a DNA/HDAC dual-targeting inhibitor. Vorambucil obtained both potent DNA and HDACs inhibitory activities. Molecular docking results supported the initial pharmacophoric hypothesis and rationalized the potent inhibitory activity of vorambucil against HDAC1, HDAC2 and HDAC6. Vorambucil showed potent antiproliferative activity against all the test four cancer cell lines with IC50 values of as low as 3.2-6.2µM and exhibited 5.0-18.3-fold enhanced antiproliferative activity than chlorambucil. Vorambucil also significantly inhibits colony formation of A375 cancer cells. Further investigation showed that vorambucil remarkably induced apoptosis and arrested the cell cycle of A375 cells at G2/M phase. Vorambucil could be a promising candidate and a useful tool to elucidate the role of those DNA/HDAC dual-targeting inhibitors for cancer therapy.
Assuntos
Antineoplásicos/farmacologia , Clorambucila/farmacologia , DNA/efeitos dos fármacos , Desenho de Fármacos , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/metabolismo , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores de Histona Desacetilases/síntese química , Inibidores de Histona Desacetilases/química , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
Multidrug resistance (MDR) limits the application of a large number of cancer-fighting agents in clinical therapy. One reason is that P-glycoprotein (Pgp) efflux pumps are usually overexpressed and lead to drug efflux in the cancer cells, which limits the viability of many chemotherapeutics. Current available inhibitors which block the Pgp pump efflux are usually not widely used in clinical practice, because they change other drug pharmacokinetic profiles or increase side effects. Here, through covalent linkage of cancer-targeting delocalized lipophilic cation FF and DNA-damaging drug nitrogen mustard chlorambucil (CLB), we rationally designed and synthesized a tumor-targeting anticancer agent FFCLB. And we found and proved that the FFCLB was capable of reducing the outflow of Pgp substrates efficiently. This conjugate selectively improves adriamycin uptake and toxicity through reducing MDR1 mRNA and Pgp protein expression. Based on molecular targeted strategy, this study can facilitate the discovery of superior MDR reducing agents to provide a more effective and safer way of resensitizing MDR.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Clorambucila/farmacologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Cátions/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Camundongos , Simulação de Acoplamento Molecular , Estrutura Molecular , Células NIH 3T3 , Relação Estrutura-AtividadeRESUMO
Glycosylated antitumor ether lipids (GAELs) kill cancer cells and cancer stem cells via a novel, apoptosis-independent mechanism. In contrast, chlorambucil, a drug in clinical use for the treatment of chronic lymphocytic leukemia reacts with nucleophiles within the major groove of DNA, leading to apoptosis. We hypothesized that hybrid molecules that combine apoptosis-dependent and apoptosis-independent mode of actions in a single molecule may lead to enhanced antitumor activity. Here, we describe the antitumor activities of chlorambucil-linked glucosamine-derived glycerolipid hybrids and investigate the role of the chlorambucil moiety and the effect of cationic charge on the hybrid molecule. Three hybrids and two control GAELs were synthesized and their activities against breast (JIMT1, MDA-MB-231, BT474), pancreas (MiaPaCa2) and prostate (DU145, PC3) cancer cell lines were determined using MTS assay. Hybrid 3 displayed the most potent activity on DU145 at CC50 of 6.0 µM while hybrid 4 displayed the best activity on JIMT1 at 7.5 µM. Hybrid 5 exhibited no activity at the highest concentration tested (CC50 >20 µM), underscoring the significance of the cationic charge at C-2 position as previously reported. Although chlorambucil (2) itself showed very little activity against all the six cell lines (CC50 >150 µM), GAELs 6 and 7 which lack the chlorambucil moiety were consistently less active than 3 and 4, suggesting that the chlorambucil moiety contributes to the overall activity. The hybrids were however not as active as the parent GAEL 1 against MiaPaCa2 whereas 6 restored activity comparable to 1.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Clorambucila/química , Clorambucila/farmacologia , Desenho de Fármacos , Éteres/química , Glicolipídeos/química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Éteres/farmacologia , Glicolipídeos/farmacologia , Glicosilação , Humanos , Estrutura Molecular , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
A newsworthy class of carboxylate esters based on the (benzo[a]acridin-12-yl)methyl (BAM) chromophore has been shown to perform dual functions as a "pH sensitive fluorescent probe" and a "phototrigger" for acids. The photophysical properties of all the BAM ester conjugates were investigated and found to be highly sensitive to solvent polarity, H-bonding capability and pH of the environment. On irradiation using UV light (≥410 nm), BAM ester conjugates underwent heterolytic cleavage of C-O bonds resulting in efficient release of carboxylic and amino acids. Interestingly, the newly synthesized BAM chromophore was also explored for the construction of a drug delivery system (DDS). In the current DDS, the BAM chromophore plays two important roles: (i) a "fluorophore" for cell imaging and (ii) a "phototrigger" for the drug release. In vitro biological studies revealed that the newly developed BAM based DDS has a good biocompatibility, cellular uptake properties and efficient photoregulated anticancer drug release ability.
Assuntos
Acridinas/síntese química , Acridinas/farmacologia , Ésteres/síntese química , Ésteres/farmacologia , Corantes Fluorescentes/síntese química , Luz , Processos Fotoquímicos , Acridinas/química , Antineoplásicos/farmacologia , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Clorambucila/farmacologia , Sistemas de Liberação de Medicamentos , Ésteres/química , Corantes Fluorescentes/química , Células HeLa , Humanos , Ligação de Hidrogênio/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Simulação de Acoplamento Molecular , Imagem Molecular , Fotólise/efeitos dos fármacos , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Fatores de TempoRESUMO
Nitrogen mustards, widely used as chemotherapeutics, have limited safety and efficacy. Mitochondria lack a functional nucleotide excision repair mechanism to repair DNA adducts and are sensitive to alkylating agents. Importantly, cancer cells have higher intrinsic mitochondrial membrane potential (Δψmt) than normal cells. Therefore, selectively targeting nitrogen mustards to cancer cell mitochondria based on Δψmt could overcome those limitations. Herein, we describe the design, synthesis, and evaluation of Mito-Chlor, a triphenylphosphonium derivative of the nitrogen mustard chlorambucil. We show that Mito-Chlor localizes to cancer cell mitochondria where it acts on mtDNA to arrest cell cycle and induce cell death, resulting in a 80-fold enhancement of cell kill in a panel of breast and pancreatic cancer cell lines that are insensitive to the parent drug. Significantly, Mito-Chlor delayed tumor progression in a mouse xenograft model of human pancreatic cancer. This is a first example of repurposing chlorambucil, a drug not used in breast and pancreatic cancer treatment, as a novel drug candidate for these diseases.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/patologia , Clorambucila/farmacologia , Mitocôndrias/efeitos dos fármacos , Neoplasias Pancreáticas/patologia , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/metabolismo , Transporte Biológico , Proliferação de Células/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Clorambucila/metabolismo , Quebras de DNA de Cadeia Dupla/efeitos dos fármacos , DNA Mitocondrial/genética , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Histonas/metabolismo , Humanos , Camundongos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Fosforilação/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
A new polyfluorene derivative containing pendent alkylating chlorambucil (PFP-Cbl) was synthesized and characterized. Under direct incubation with DNA in vitro, PFP-Cbl could undergo an efficient DNA alkylating reaction and induce DNA cross-linking. In vitro transcription and translation experiment exhibited that the PFP-Cbl significantly down-regulated the gene expression of luciferase reporter plasmid. The down-regulation of gene expression was also verified through the transfection experiment of p-EGFP plasmid, which showed decreased green fluorescent protein (GFP) in cells. Meanwhile, the self-luminous property of PFP-Cbl could make it able to trace the internalized PFP-Cbl and plasmid complexes resulted from cross-linking in cells by fluorescent microscopy. Combining the features of alkylating function, multivalent binding sites, and fluorescent characteristics, PFP-Cbl provides a new insight in the area of gene regulation and extends the new applications of conjugated polymers (CPs).
Assuntos
DNA/metabolismo , Regulação da Expressão Gênica , Plasmídeos/metabolismo , Polímeros/síntese química , Alquilação , Antineoplásicos Alquilantes/síntese química , Sítios de Ligação/genética , Clorambucila/síntese química , Reagentes de Ligações Cruzadas/síntese química , DNA/genética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Humanos , Microscopia de Fluorescência , Plasmídeos/genética , Polímeros/metabolismoRESUMO
We have successfully delivered a reactive alkylating agent, chlorambucil (Cbl), to the mitochondria of mammalian cells. Here, we characterize the mechanism of cell death for mitochondria-targeted chlorambucil (mt-Cbl) in vitro and assess its efficacy in a xenograft mouse model of leukemia. Using a ρ° cell model, we show that mt-Cbl toxicity is not dependent on mitochondrial DNA damage. We also illustrate that re-targeting Cbl to mitochondria results in a shift in the cell death mechanism from apoptosis to necrosis, and that this behavior is a general feature of mitochondria-targeted Cbl. Despite the change in cell death mechanisms, we show that mt-Cbl is still effective in vivo and has an improved pharmacokinetic profile compared to the parent drug. These findings illustrate that mitochondrial rerouting changes the site of action of Cbl and also alters the cell death mechanism drastically without compromising in vivo efficacy. Thus, mitochondrial delivery allows the exploitation of Cbl as a promiscuous mitochondrial protein inhibitor with promising therapeutic potential.
Assuntos
Antineoplásicos Alquilantes/farmacologia , Peptídeos Penetradores de Células/química , Clorambucila/farmacologia , Sistemas de Liberação de Medicamentos/métodos , Leucemia/tratamento farmacológico , Mitocôndrias/efeitos dos fármacos , Animais , Antineoplásicos Alquilantes/síntese química , Apoptose/efeitos dos fármacos , Clorambucila/síntese química , Reagentes de Ligações Cruzadas/química , DNA Mitocondrial , Células HeLa , Humanos , Leucemia/metabolismo , Leucemia/patologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Necrose/patologia , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Cancer is one of the most fatal diseases in the world and it has been years that finding new drugs and chemotherapeutic techniques with lowest side effects become one of the most important challenging matters needs really hard efforts. Chlorambucil (CBL), an ancient direct-acting alkylating anticancer agent, is commonly used for initial treatment of some kinds of cancers but the use of CBL is often limited because of the unpleasant side effects due to its lack of specificity for targeting cancer cells. In this research we tried to increase the specificity of CBL by producing a novel conjugate by using glutamine amino acid (Glut). Based on previous studies, poly amines and nitrogen compounds noticeably are used by cancer cells increasingly; therefore we decided to increase the efficiency and specificity of CBL by designing and producing a novel anti cancer conjugate using glutamine amino acid as an uptake enhancer, CBL, and Adipic acid Dihydrazide (ADH) as a spacer and linker. The biological tests were carried out on HT29 colorectal cancer cell line to evaluate its anticancer properties. Biological tests like MTT assay, finding IC50, evaluating the induced mechanism of the death of our novel CBL-Glutamine conjugate on HT29 cells, testing abnormal toxicity of this conjugate on mice in comparison with CBL drug were careid out. We found that not only CBL-Glutamine conjugate preserved its anti cancer property with regard to CBL drug, but also it represent lower abnormal toxicity in mice. Apoptosis was detected as its mechanism of the death. Our present study provides a promising strategy for targeting cancer cells using amino acids nano-conjugate drugs. The future perspectives have also been highlighted in continuing similar and relative researches.
Assuntos
Adipatos/farmacologia , Antineoplásicos/farmacologia , Clorambucila/análogos & derivados , Neoplasias Colorretais/tratamento farmacológico , Glutamina/análogos & derivados , Terapia de Alvo Molecular , Adipatos/síntese química , Adipatos/química , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Clorambucila/síntese química , Clorambucila/química , Clorambucila/farmacologia , Neoplasias Colorretais/patologia , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Glutamina/síntese química , Glutamina/química , Glutamina/farmacologia , Células HT29 , Humanos , Camundongos , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
The amino analogues of pentamidine with a polymethylene (n = 3 - 6) chain and their chlorambucil derivatives were synthesized. The obtained compounds revealed cytotoxic effect on MCF-7 human breast cancer cell line (IC50 = 22 - 95 +/- 2 pM), mainly by the induction of apoptosis. The topoisomerase I/II inhibition assay and the ethidium displacement assay with the use of pBR322 plasmid DNA were used to the study of mechanism by which the obtained compounds could act. All the compounds are able to bind with DNA and interfere in vitro with the activity of topoisomerase (I and II). The determination of association constants with the use of calf thymus DNA, T4 coliphage DNA, poly(dA-dT)2 and poly(dG-dC)2 showed that the tested compounds bind within minor groove of B-DNA, but not selectively. The alkylating activity of chlorambucil derivatives determined in vitro using a Preussmann test was similar to the activity of chlorambucil. The influence of all the compounds on the amidolytic activity of plasmin and trypsin was also examined. The plasmin activity was inhibited by pentamidine, chlorambucil and aromatic bis-amines (IC50 = 0.1 - 8 mM), whereas the trypsin activity was influenced only by pentamidine.