ABSTRACT
COVID-19 is an ongoing worldwide pandemic. Even today, there is a need for the development of effective therapeutic agents. SARS-CoV-2 is known as the causative virus of COVID-19, and its main protease is one of the enzymes essential for its growth and is considered a drug discovery target. In this study, we evaluated the inhibitory activities of a variety of fullerene derivatives, including newly synthesized derivatives, against the main protease of SARS-CoV-2. As a result, the malonic acid-type fullerene derivatives showed the strongest inhibitory activity.
Subject(s)
COVID-19 , Fullerenes , Humans , SARS-CoV-2 , Fullerenes/pharmacology , Protease Inhibitors/pharmacology , Antiviral Agents/pharmacology , Coronavirus 3C Proteases , Molecular Docking SimulationABSTRACT
In the treatment of breakpoint cluster region-Abelson (BCR-ABL)-positive chronic myeloid leukemia (CML) using BCR-ABL inhibitors, the appearance of a gatekeeper mutation (T315I) in BCR-ABL is a serious issue. Therefore, the development of novel drugs that overcome acquired resistance to BCR-ABL inhibitors by CML cells is required. We previously demonstrated that a bis-pyridinium fullerene derivative (BPF) induced apoptosis in human chronic myeloid leukemia (CML)-derived K562 cells partially through the generation of reactive oxygen species (ROS). We herein show that BPF enhanced the activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase-extracellular signal-regulated kinase (MEK-ERK) pathway in a ROS-independent manner. BPF-induced apoptosis was attenuated by trametinib, suggesting the functional involvement of the MEK-ERK pathway in apoptosis in K562 cells. In addition, the constitutive activation of the MEK-ERK pathway by the enforced expression of the BRAFV600E mutant significantly increased the sensitivity of K562 cells to BPF. These results confirmed for the first time that BPF induces apoptosis in K562 cells through dual pathways-ROS production and the activation of the MEK-ERK pathway. Furthermore, BPF induced cell death in transformed Ba/F3 cells expressing not only BCR-ABL but also T315I mutant through the activation of the MEK-ERK pathway. These results indicate that BPF is as an effective CML drug that overcomes resistance to BCR-ABL inhibitors.
Subject(s)
Apoptosis/drug effects , Fullerenes/pharmacology , Fusion Proteins, bcr-abl/genetics , MAP Kinase Signaling System/drug effects , Reactive Oxygen Species/metabolism , Animals , Fullerenes/chemistry , Genes, Essential , Humans , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Mice , Models, Biological , Mutation , Proto-Oncogene Proteins B-raf/genetics , Proto-Oncogene Proteins B-raf/metabolismABSTRACT
Acyl glucuronides are common metabolites of carboxylic acid-containing compounds. Since acyl glucuronides sometimes show high reactivity, they are considered to be involved in drug toxicity. Therefore, it is important to evaluate the risk posed by acyl glucuronides in the development of safe drugs; however, there are no suitable evaluation methods for the early stages of drug discovery. We aimed to develop a trapping reagent that detects reactive acyl glucuronides to assess their risk. We designed a diamine-structured trapping reagent, Dap-Dan, and compared its trapping ability with the reported one that has an amino group, and results showed that Dap-Dan showed higher accuracy. In the trapping assay with 17 medicines containing a carboxylic acid, Dap-Dan trapped acyl glucuronides that had a higher risk of toxicity. In conclusion, Dap-Dan can be useful for evaluating the risk of reactive acyl glucuronides.
Subject(s)
Fluorescent Dyes/chemistry , Glucuronides/analysis , Fluorescent Dyes/chemical synthesis , Glucuronides/metabolism , Humans , Molecular StructureABSTRACT
In the present study, we newly synthesized four types of novel fullerene derivatives: pyridinium/ethyl ester-type derivatives 3b-3l, pyridinium/carboxylic acid-type derivatives 4a, 4e, 4f, pyridinium/amide-type derivative 5a, and pyridinium/2-morpholinone-type derivative 6a. Among the assessed compounds, cis-3c, cis-3d, trans-3e, trans-3h, cis-3l, cis-4e, cis-4f, trans-4f, and cis-5a were found to inhibit HIV-1 reverse transcriptase (HIV-RT), HIV-1 protease (HIV-PR), and HCV NS5B polymerase (HCV NS5B), with IC50 values observed in the micromolar range. Cellular uptake of pyridinium/ethyl ester-type derivatives was higher than that of corresponding pyridinium/carboxylic acid-type derivatives and pyridinium/amide-type derivatives. This result might indicate that pyridinium/ethyl ester-type derivatives are expected to be lead compounds for multitargeting drugs to treat HIV/HCV coinfection.
Subject(s)
Anti-HIV Agents/pharmacology , Fullerenes/pharmacology , HIV Protease Inhibitors/pharmacology , Pyridinium Compounds/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , Viral Nonstructural Proteins/antagonists & inhibitors , Animals , Anti-HIV Agents/chemical synthesis , Anti-HIV Agents/toxicity , Cell Line, Tumor , Fullerenes/chemistry , Fullerenes/toxicity , HIV Protease/metabolism , HIV Protease Inhibitors/chemical synthesis , HIV Protease Inhibitors/toxicity , HIV Reverse Transcriptase/antagonists & inhibitors , HIV-1/enzymology , Hepacivirus/enzymology , Humans , Mice , Molecular Structure , NIH 3T3 Cells , Pyridinium Compounds/chemical synthesis , Pyridinium Compounds/toxicity , Reverse Transcriptase Inhibitors/chemical synthesis , Reverse Transcriptase Inhibitors/toxicity , Structure-Activity RelationshipABSTRACT
In the present study, we newly synthesized three types of novel fullerene derivatives: pyridinium-type derivatives trans-3a and 4a-5b, piperidinium-type derivative 9, and proline-type derivatives 10a-12. Among the assessed compounds, 5a, 10e, 10f, 10i, 11a-d, and 12 were found to inhibit both HIV reverse transcriptase and HIV protease (HIV-PR), with IC50 values in the low micromolar range being observed. Regarding HIV-PR inhibition activity, proline-type derivatives 11a-11d and 12, bearing an alkyl chain between the hydroxylmethylcarbonyl (HMC) moiety and pyrrolidine ring, were more potent than other derivatives. This result might indicate that connecting HMC moieties with proline-type fullerene derivatives through properly sized alkyl chain leads to improved HIV-PR inhibitory activity.
Subject(s)
Fullerenes/pharmacology , HIV Protease Inhibitors/pharmacology , HIV Protease/metabolism , HIV Reverse Transcriptase/antagonists & inhibitors , Pyridinium Compounds/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , Dose-Response Relationship, Drug , Fullerenes/chemistry , HIV Protease Inhibitors/chemical synthesis , HIV Protease Inhibitors/chemistry , HIV Reverse Transcriptase/metabolism , Molecular Structure , Pyridinium Compounds/chemistry , Reverse Transcriptase Inhibitors/chemical synthesis , Reverse Transcriptase Inhibitors/chemistry , Structure-Activity RelationshipABSTRACT
We synthesized six novel BBR derivatives that were designed to avoid metabolic activation via ipso-substitution and evaluated for their degree of toxicity and hURAT1 inhibition. It was found that all of the derivatives demonstrate lower cytotoxicity in mouse hepatocytes and lower levels of metabolic activation than BBR, while maintaining their inhibitory activity toward the uric acid transporter. We propose that these derivatives could serve as effective uricosuric agents that have much better safety profiles than BBR.
Subject(s)
Benzbromarone/analogs & derivatives , Benzbromarone/metabolism , Organic Anion Transporters/antagonists & inhibitors , Organic Cation Transport Proteins/antagonists & inhibitors , Uricosuric Agents/chemistry , Uricosuric Agents/metabolism , Activation, Metabolic , Animals , Benzbromarone/pharmacology , Benzbromarone/toxicity , Chemistry Techniques, Synthetic , HEK293 Cells , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Mice , Microsomes, Liver/drug effects , Microsomes, Liver/metabolism , Mitochondria/drug effects , Mitochondria/metabolism , Organic Anion Transporters/metabolism , Organic Cation Transport Proteins/metabolism , Rats , Uric Acid/metabolism , Uricosuric Agents/pharmacology , Uricosuric Agents/toxicityABSTRACT
The Keap1-Nrf2 system is an attractive target for drug discovery regarding various unmet medical needs. Only covalent inhibitors for protein-protein interaction (PPI) between Keap1 and Nrf2 to activate Nrf2 have been approved or are under clinical trials, but such electrophilic compounds lack selectivity. Therefore, specific non-covalent Keap1-Nrf2 PPI inhibitors are expected to be safer Nrf2 activators. We found a novel class of non-covalent Keap1-Nrf2 PPI inhibitor that has a benzo[g]indole skeleton and an indole-3-hydroxamic acid moiety and that exhibits significant PPI inhibitory activity. Additionally, the benzo[g]indole-3-carbohydrazide derivatives were newly prepared. The benzo[g]indole derivatives showed a stronger Keap1-Nrf2 PPI inhibitory activity than Cpd16, a previously reported non-covalent PPI inhibitor. Moreover, most of the PPI inhibitors showed a high metabolic stability in a human microsome system with a low cytotoxicity against HepG2 cell lines, which suggests that novel benzo[g]indole-type Keap1-Nrf2 PPI inhibitors are expected to be biological tools or lead compounds for Nrf2 activators.
Subject(s)
Indoles/chemistry , Kelch-Like ECH-Associated Protein 1/metabolism , NF-E2-Related Factor 2/metabolism , Cell Survival/drug effects , Drug Evaluation, Preclinical , Hep G2 Cells , Humans , Hydroxamic Acids/chemical synthesis , Hydroxamic Acids/chemistry , Hydroxamic Acids/toxicity , Indoles/chemical synthesis , Indoles/toxicity , Inhibitory Concentration 50 , Kelch-Like ECH-Associated Protein 1/antagonists & inhibitors , Microsomes, Liver/metabolism , NF-E2-Related Factor 2/antagonists & inhibitors , Protein Interaction Domains and MotifsABSTRACT
We evaluated the Hepatitis C virus (HCV) NS5B polymerase and HCV NS3/4A protease inhibition activities of a new set of proline-type fullerene derivatives. All of the compounds had the potential to inhibit both the enzymes, indicating that the fullerene derivatives may be dual inhibitors against NS5B and NS3/4A and could be novel lead compounds for the treatment of HCV infections.
Subject(s)
Fullerenes/pharmacology , Hepacivirus/enzymology , Viral Nonstructural Proteins/antagonists & inhibitors , Inhibitory Concentration 50 , Mass Spectrometry , Proton Magnetic Resonance SpectroscopyABSTRACT
The Keap1-Nrf2 system is involved not only in biological defense but also in malignancy progression and chemoresistance. The ubiquitin-binding protein p62/Sqstm1 (p62), which is highly expressed in several cancers, competes with Nrf2 for Keap1 binding, leading to activation of Nrf2-mediated gene expression and survival of cancer cells. We had previously identified an inhibitor for the Keap1-phosphorylated-p62 (p-p62) protein-protein interaction (PPI), the acetonyl naphthalene derivative K67. In this study, we established facile synthetic routes for K67 and derivatives with various side chains on the C-2 position of naphthalene ring. K67 possessed high selectivity in the inhibition of Keap1-p-p62. Other derivatives showed potent Keap1-Nrf2 and Keap1-p-p62 PPI inhibitory activities, though the selectivity between the two activities was lower than K67.
Subject(s)
1-Naphthylamine/analogs & derivatives , Kelch-Like ECH-Associated Protein 1/antagonists & inhibitors , NF-E2-Related Factor 2/antagonists & inhibitors , Naphthalenes/pharmacology , RNA-Binding Proteins/antagonists & inhibitors , Sulfonamides/pharmacology , ras GTPase-Activating Proteins/antagonists & inhibitors , 1-Naphthylamine/chemical synthesis , 1-Naphthylamine/chemistry , 1-Naphthylamine/pharmacology , Dose-Response Relationship, Drug , Humans , Kelch-Like ECH-Associated Protein 1/chemistry , Molecular Structure , NF-E2-Related Factor 2/chemistry , Naphthalenes/chemistry , Protein Binding/drug effects , RNA-Binding Proteins/chemistry , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/chemistry , ras GTPase-Activating Proteins/chemistryABSTRACT
Antioxidant treatments have been expected to be a novel therapeutics for various oxidative stress-mediated disorders. Our previous study revealed that 5-hydroxyoxindole and its 3-phenacyl-3-hydroxy derivatives showed excellent antioxidant activities such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and lipid-peroxidation inhibitory activity. However, the DPPH radical scavenging activity of the 3,3-disubstituted derivatives was lower than that of the original 5-hydroxyoxindole. In the present study, we synthesized novel 3-monosubstituted 5-hydroxyoxindole derivatives that exhibited stronger DPPH radical scavenging activities and lipid peroxidation-inhibitory activities than the 3,3-disubstituted 5-hydroxyoxindoles. Moreover, the 3-monosubstituted 5-hydroxyoxindole derivatives showed neither an iron-mediated pro-oxidant effect nor a remarkable cytotoxicity against HL-60 cell lines except some of the highly lipophilic compounds. These results indicate that 3-monosubstituted 5-hydroxyoxindoles can be used as a promising antioxidant scaffold for drug discovery.
ABSTRACT
Benzbromarone (BBR) is a hepatotoxic drug, but the detailed mechanism of its toxicity remains unknown. We identified 2,6-dibromohydroquinone (DBH) and mono-debrominated catechol (2-ethyl-3-(3-bromo-4,5-dihydroxybenzoyl)benzofuran; CAT) as novel metabolites of BBR in rat and human liver microsomal systems by comparison with chemically synthesized authentic compounds, and we also elucidated that DBH is formed by cytochrome P450 2C9 and that CAT is formed mainly by CYP1A1, 2D6, 2E1, and 3A4. Furthermore, CAT, DBH, and the oxidized form of DBH are highly cytotoxic in HepG2 compared with BBR. Taken together, our data demonstrate that DBH, a novel reactive metabolite, may be relevant to BBR-induced hepatotoxicity.
Subject(s)
Benzbromarone/pharmacokinetics , Cytochrome P-450 Enzyme System/metabolism , Activation, Metabolic , Animals , Hep G2 Cells , Humans , Microsomes, Liver/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
In the present study, we describe the synthesis of a novel set of pyridine/pyridinium-type fullerene derivatives. The products were assessed for human immunodeficiency virus-reverse transcriptase inhibition activities. All novel fullerene derivatives showed potent human immunodeficiency virus-reverse transcriptase inhibition without cytotoxicity.
Subject(s)
Anti-HIV Agents/pharmacology , Fullerenes/pharmacology , Pyridines/pharmacology , Pyridinium Compounds/pharmacology , Reverse Transcriptase Inhibitors/pharmacology , Anti-HIV Agents/chemical synthesis , Cell Survival/drug effects , Fullerenes/chemistry , HIV Reverse Transcriptase , HL-60 Cells , Humans , Pyridines/chemical synthesis , Pyridinium Compounds/chemical synthesis , Reverse Transcriptase Inhibitors/chemical synthesisABSTRACT
Primary effusion lymphoma (PEL) is a subtype of non-Hodgkin's B-cell lymphoma and is an aggressive neoplasm caused by Kaposi's sarcoma-associated herpesvirus (KSHV) in immunosuppressed patients. In general, PEL cells are derived from post-germinal center B-cells and are infected with KSHV. To evaluate potential novel anti-tumor compounds against KSHV-associated PEL, seven water-soluble fullerene derivatives were evaluated as potential drug candidates for the treatment of PEL. Herein, we discovered a pyrrolidinium fullerene derivative, 1,1,1',1'-tetramethyl [60]fullerenodipyrrolidinium diiodide, which induced apoptosis of PEL cells via a novel mechanism, the caspase-9 activation by suppressing the caspase-9 phosphorylation, causing caspase-9 inactivation. Pyrrolidinium fullerene treatment reduced significantly the viability of PEL cells compared with KSHV-uninfected lymphoma cells, and induced the apoptosis of PEL cells by activating caspase-9 via procaspase-9 cleavage. Pyrrolidinium fullerene additionally reduced the Ser473 phosphorylation of Akt and Ser196 of procaspase-9. Ser473-phosphorylated Akt (i.e., activated Akt) phosphorylates Ser196 in procaspase-9, causing inactivation of procaspase-9. We also demonstrated that Akt inhibitors suppressed the proliferation of PEL cells compared with KSHV-uninfected cells. Our data therefore suggest that Akt activation is essential for cell survival in PEL and a pyrrolidinium fullerene derivative induced apoptosis by activating caspase-9 via suppression of Akt in PEL cells. In addition, we evaluated whether pyrrolidinium fullerene in combination with the HSP90 inhibitor (geldanamycin; GA) or valproate, potentiated the cytotoxic effects on PEL cells. Compared to treatment with pyrrolidinium fullerene alone, the addition of low-concentration GA or valproate enhanced the cytotoxic activity of pyrrolidinium fullerene. These results indicate that pyrrolidinium fullerene could be used as a novel therapy for the treatment of PEL.
Subject(s)
Antineoplastic Agents/pharmacology , Bridged-Ring Compounds/pharmacology , Lymphoma, Primary Effusion/drug therapy , Lymphoma, Primary Effusion/pathology , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Pyrrolidines/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Apoptosis/drug effects , Benzoquinones/administration & dosage , Benzoquinones/pharmacology , Caspase 9/metabolism , Caspase Inhibitors/pharmacology , Cell Line, Tumor , Drug Screening Assays, Antitumor/methods , Fullerenes/chemistry , Fullerenes/pharmacology , HSP90 Heat-Shock Proteins/antagonists & inhibitors , Herpesvirus 8, Human/pathogenicity , Humans , Lactams, Macrocyclic/administration & dosage , Lactams, Macrocyclic/pharmacology , Lymphoma, Primary Effusion/metabolism , Lymphoma, Primary Effusion/virology , Phosphorylation/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Serine/metabolism , Virion/drug effectsABSTRACT
The antioxidant activities of 5-hydroxyoxindole (1) and newly synthesized 3,5-dihydroxy-3-phenacyl-2-oxindole derivatives against rat liver microsome/tert-butylhydroperoxide system-induced lipid peroxidation and hydrogen peroxide-induced intracellular oxidative stress were investigated. Compound 1 and its derivatives showed significant suppression of lipid peroxidation and an intracellular oxidative stress. The effects of the more lipophilic derivatives tended to be greater than that of the original compound 1. The cytotoxicity of all of the oxindole derivatives on human promyelocytic leukemia HL60 cells was lower than that of 2,6-di(tert-butyl)-4-hydroxytoluene (BHT), a widely used phenolic antioxidant. These results show that compound 1 and its 3-substituted derivatives could be good lead candidates for future novel antioxidant therapeutics.
Subject(s)
Antineoplastic Agents/pharmacology , Antioxidants/pharmacology , Indoles/pharmacology , Lipid Peroxidation/drug effects , Oxidative Stress/drug effects , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Antioxidants/chemical synthesis , Antioxidants/chemistry , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , HL-60 Cells , Humans , Hydrogen Peroxide/antagonists & inhibitors , Hydrogen Peroxide/pharmacology , Indoles/chemical synthesis , Indoles/chemistry , Microsomes, Liver/drug effects , Microsomes, Liver/metabolism , Molecular Structure , Oxindoles , Rats , Structure-Activity Relationship , Tumor Cells, Cultured , tert-Butylhydroperoxide/antagonists & inhibitors , tert-Butylhydroperoxide/pharmacologyABSTRACT
Although acyl-CoA conjugates are known to have higher reactivity than acyl glucuronides, few studies have been conducted to evaluate the risk of the conjugates. In the present study, we aimed to develop a trapping assay for acyl-CoA conjugates using trapping reagents we have developed previously. It was revealed that Cys-Dan, which has both a thiol and an amino group, was the most effective in forming stable adducts containing an amide bond after intramolecular acyl migration. Additionally, we also developed a hepatocyte-based trapping assay in the present study to overcome the shortcomings of liver microsomes. Although liver microsomes are commonly used as enzyme sources in trapping assays, they lack some of the enzymes required for drug metabolism and detoxification systems. In human hepatocytes, our three trapping reagents, CysGlu-Dan, Dap-Dan and Cys-Dan, captured CYP-dependent reactive metabolites, reactive acyl glucuronides, and reactive acyl-CoA conjugates, respectively. The work suggests that the trapping assay with the reagents in hepatocytes is useful to evaluate the risk of reactive metabolites in drug discovery.
Subject(s)
Acyl Coenzyme A , Glucuronides , Humans , Acyl Coenzyme A/chemistry , Acyl Coenzyme A/metabolism , Acyl Coenzyme A/pharmacology , Indicators and Reagents/metabolism , Glucuronides/metabolism , Microsomes, Liver/metabolism , Sulfhydryl Compounds/metabolism , Sulfhydryl Compounds/pharmacologyABSTRACT
It has been demonstrated that the function of mammalian clock gene transcripts is controlled by the binding of heme in vitro. To examine the effects of heme on biological rhythms in vivo, we measured locomotor activity (LA) and core body temperature (T(b)) in a mouse model of porphyria with impaired heme biosynthesis by feeding mice a griseofulvin (GF)-containing diet. Mice fed with a 2.0% GF-containing diet (GF2.0) transiently exhibited phase advance or phase advance-like phenomenon by 1-3 h in terms of the biological rhythms of T(b) or LA, respectively (both, P < 0.05) while mice were kept under conditions of a light/dark cycle (12 h:12 h). We also observed a transient, ~0.3 h shortening of the period of circadian T(b) rhythms in mice kept under conditions of constant darkness (P < 0.01). Interestingly, the observed duration of abnormal circadian rhythms in GF2.0 mice lasted between 1 and 3 wk after the onset of GF ingestion; this finding correlated well with the extent of impairment of heme biosynthesis. When we examined the effects of therapeutic agents for acute porphyria, heme, and hypertonic glucose on the pathological status of GF2.0 mice, it was found that the intraperitoneal administration of heme (10 mg·kg(-1)·day(-1)) or glucose (9 g·kg(-1)·day(-1)) for 7 days partially reversed (50%) increases in urinary δ-aminolevulinic acids levels associated with acute porphyria. Treatment with heme, but not with glucose, suppressed the phase advance (-like phenomenon) in the diurnal rhythms (P < 0.05) and restored the decrease of heme (P < 0.01) in GF2.0 mice. These results suggest that impairments of heme biosynthesis, in particular a decrease in heme, may affect phase and period of circadian rhythms in animals.
Subject(s)
Body Temperature/physiology , Circadian Rhythm/physiology , Heme/metabolism , Porphyrias/metabolism , Animals , Body Temperature/drug effects , Circadian Rhythm/drug effects , Disease Models, Animal , Glucose/pharmacology , Griseofulvin/adverse effects , Griseofulvin/pharmacology , Heme/pharmacology , Locomotion/drug effects , Locomotion/physiology , Male , Mice , Mice, Inbred ICR , Porphyrias/chemically induced , Porphyrias/physiopathologyABSTRACT
Fullerene (C60) and fullerene derivatives are attractive novel compounds not only for carbon materials of nanotechnology but also for medical fields because of its unique chemical and physical properties. We intend to develop fullerene derivatives as novel lead compounds for drug discovery. At first, we synthesized many types of water-soluble fullerene derivatives to investigate their biological activities because of their poor solubility in water. We found that anionic fullerene derivatives possess anti-oxidant activities, whereas di-cationic fullerene derivatives exhibited antiproliferative activities against various cancer cell lines including drug-resistant cells. Proline-type fullerene derivatives showed inhibitory activities against human immunodeficiency virus (HIV) reverse transcriptase, HIV protease, hepatitis C virus (HCV) NS5B RNA polymerase, and HCV NS3/4A protease. These activities may strongly inhibit virus replication via a synergistic effect and fullerene derivatives may be used as novel multi-target drugs for the treatment of AIDS and hepatitis C in the future.
Subject(s)
Drug Discovery/methods , Fullerenes , Acquired Immunodeficiency Syndrome/drug therapy , Antineoplastic Agents , Antioxidants , Antiviral Agents , Fullerenes/chemistry , Fullerenes/pharmacology , Fullerenes/therapeutic use , Hepatitis C/drug therapy , Humans , Solubility , Virus Replication/drug effects , WaterABSTRACT
A fusion protein, Breakpoint cluster region-Abelson (BCR-ABL) is responsible for the development of chronic myeloid leukemia (CML) and acute lymphocytic leukemia (ALL). Inhibitors against BCR-ABL are effective for the treatment of leukemia; however, a gatekeeper mutation (T315I) in BCR-ABL results in resistance to these inhibitors, which markedly impedes their efficacy. We herein demonstrated that a bis-pyridinium fullerene derivative (BPF) significantly induced apoptosis in human CML-derived K562 cells and ALL-derived SUP-B15 cells via the generation of reactive oxygen species (ROS). BPF reduced the expression of Bcr-Abl mRNA by inhibiting expression of c-Myc through ROS production. BPF also accelerated protein degradation of BCR-ABL through ROS production. Furthermore, BPF down-regulated the expression of not only BCR-ABL but also T315I-mutated BCR-ABL in ROS-dependent manner. As a result, BPF effectively induced apoptosis in transformed Ba/F3 cells expressing both BCR-ABL and T315I-mutated BCR-ABL. Collectively, these results indicate the potential of BPF as an effective leukemia drug that overcomes resistance to BCR-ABL inhibitors.
Subject(s)
Antineoplastic Agents , Fullerenes , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Apoptosis , Drug Resistance, Neoplasm , Fullerenes/chemistry , Fusion Proteins, bcr-abl/metabolism , Humans , Imatinib Mesylate/pharmacology , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Reactive Oxygen Species/metabolismABSTRACT
Tofacitinib (TFT), a JAK inhibitor used for the treatment of rheumatoid arthritis and other diseases, is associated with severe liver injury that is believed to be caused by its reactive aldehyde or epoxide metabolites. In this study, we synthesized six tofacitinib analogs designed to avoid the formation of reactive metabolites and evaluated their JAK3 inhibitory activity, metabolic stability, CYP3A time-dependent inhibition, and cytotoxicity. Our data indicated that purine analog 3, which showed little inhibition of CYP3A and cytotoxicity and inhibited JAK3 in the nanomolar range, could be a safer drug candidate than TFT. In addition, the results of the bioactivation study using TFT and its analogs suggest that the epoxide metabolite might contribute to TFT-induced CYP3A4 mechanism-based inhibition and hepatic toxicity.
Subject(s)
Piperidines , Pyrimidines , Activation, Metabolic , Cytochrome P-450 CYP3A/metabolism , Microsomes, Liver/metabolism , Piperidines/metabolism , Piperidines/pharmacology , Pyrimidines/metabolism , Pyrimidines/pharmacologyABSTRACT
Primary effusion lymphoma (PEL) is defined as a rare subtype of nonHodgkin's B cell lymphoma, which is caused by Kaposi's sarcomaassociated herpesvirus (KSHV) in immunosuppressed patients. PEL is an aggressive type of lymphoma and is frequently resistant to conventional chemotherapeutics. Therefore, the discovery of novel drug candidates for the treatment of PEL is of utmost importance. In order to discover potential novel antitumor compounds against PEL, the authors previously developed a pyrrolidiniumtype fullerene derivative, 1,1,1',1'tetramethyl [60]fullerenodipyrrolidinium diiodide (derivative #1), which induced the apoptosis of PEL cells via caspase9 activation. In the present study, the growth inhibitory effects of pyrrolidiniumtype (derivatives #1 and #2), pyridiniumtype (derivatives #3 and #5 to #9) and aniliniumtype fullerene derivatives (derivative #4) against PEL cells were evaluated. This analysis revealed a pyridiniumtype derivative (derivative #5; 3â5'(etho xycarbonyl)1',5'dihydro2'H[5,6]fullerenoC60Ih[1,9c]pyrrol2'yl]1methylpyridinium iodide), which exhibited antitumor activity against PEL cells via the downregulation of Wnt/ßcatenin signaling. Derivative #5 suppressed the viability of KSHVinfected PEL cells compared with KSHVuninfected Blymphoma cells. Furthermore, derivative #5 induced the destabilization of ßcatenin and suppressed ßcateninTCF4 transcriptional activity in PEL cells. It is known that the constitutive activation of Wnt/ßcatenin signaling is essential for the growth of KSHVinfected cells. The Wnt/ßcatenin activation in KSHVinfected cells is mediated by KSHV latencyassociated nuclear antigen (LANA). The data demonstrated that derivative #5 increased ßcatenin phosphorylation, which resulted in ßcatenin polyubiquitination and subsequent degradation. Thus, derivative #5 overcame LANAmediated ßcatenin stabilization. Furthermore, the administration of derivative #5 suppressed the development of PEL cells in the ascites of SCID mice with tumor xenografts derived from PEL cells. On the whole, these findings provide evidence that the pyridiniumtype fullerene derivative #5 exhibits antitumor activity against PEL cells in vitro and in vivo. Thus, derivative #5 may be utilized as a novel therapeutic agent for the treatment of PEL.