ABSTRACT
BACKGROUND: The skin microbiome, a diverse community of microorganisms, plays a crucial role in maintaining skin health. Among these microorganisms, the gram-positive bacterium Micrococcus luteus exhibits potential for promoting skin health. This study focuses on postbiotics derived from M. luteus YM-4, a strain isolated from human skin. OBJECTIVE: Our objective is to explore the beneficial effects of YM-4 culture filtrate on dermatological health, including enhancing barrier function, modulating immune response, and aiding recovery from environmental damage. METHODS: The effects of the YM-4 culture filtrate were tested on human keratinocytes and fibroblasts under various conditions using real-time PCR for gene expression analysis and fibroblast migration assays. A dehydration-simulated model was employed to prepare RNA-Seq samples from HaCaT cells treated with the YM-4 culture filtrate. Differentially expressed genes were identified and functionally classified through k-means clustering, gene ontology terms enrichment analyses, and protein-protein interactions mapping. RESULTS: The YM-4 culture filtrate enhanced the expression of genes involved in skin hydration, hyaluronic acid synthesis, barrier function, and cell proliferation. It also reduced inflammation markers in keratinocytes and fibroblasts under stress conditions. It mitigated UVB-induced collagen degradation while promoted collagen synthesis, suggesting anti-aging properties, and accelerated wound healing processes by promoting cell proliferation and migration. RNA sequencing analysis revealed that the YM-4 culture filtrate could reverse dehydration-induced transcriptional changes towards a state similar to untreated cells. CONCLUSION: M. luteus YM-4 culture filtrate exhibits significant therapeutic potential for dermatological applications.
Subject(s)
Dehydration , Epirubicin/analogs & derivatives , Micrococcus luteus , Humans , Dehydration/metabolism , Skin/metabolism , Collagen/metabolismABSTRACT
BACKGROUND: NC-6300 is a novel epirubicin (EPI) drug conjugated polymeric micelle developed using cutting-edge micellar nanoparticle technology. The nanoparticle epirubicin conjugates EPI to a polymer via a pH-sensitive linker which enables the selective EPI release into tumor. Tumor activity was observed in a monotherapy phase Ib trial, where two of two patients with angiosarcoma achieved a partial response. To further explore the activity of NC-6300 in angiosarcoma, an expansion cohort was undertaken. METHODS: Ten patients with angiosarcoma were enrolled in the expansion cohort. Patients were dosed using the recommended dose of 150 mg/m2 intravenously (IV) once every 3 weeks. The primary endpoint was progression-free survival. RESULTS: The most common adverse events (AEs) of any grade, regardless of the causal relationship with NC-6300, were neutropenia (90%), fatigue, and thrombocytopenia (60% each) and nausea (50%). The most common grades 3 and 4 AEs were neutropenia (80%), thrombocytopenia (40%), and anemia and leukopenia (20% each). The median progression-free survival (mPFS) for all subjects was 5.4 months. The mPFS was 3.8 months in subjects with prior anthracycline treatment and 8.2 months in subjects without prior anthracycline treatment. CONCLUSION: NC-6300 was well tolerated, showing promising activity in angiosarcoma patients without prior anthracycline treatment. NC-6300 warrants further investigation (ClinicalTrials.gov Identifier: NCT03168061).
Subject(s)
Hemangiosarcoma , Nanoparticles , Neutropenia , Thrombocytopenia , Anthracyclines , Antibiotics, Antineoplastic/therapeutic use , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Epirubicin/adverse effects , Epirubicin/analogs & derivatives , Hemangiosarcoma/chemically induced , Hemangiosarcoma/drug therapy , Humans , Micelles , Neutropenia/chemically induced , Polymers , Proteins , Thrombocytopenia/chemically inducedABSTRACT
PURPOSE: NC-6300 is a novel nanoparticle formulation of epirubicin that has a pH-sensitive linker conjugated to epirubicin. It exhibits selective tumor accumulation owing to enhanced permeability and retention effect. We conducted a phase 1b trial to determine MTD and recommended phase II dose (RP2D) of NC-6300 monotherapy in advanced, metastatic, or unresectable solid tumors, including soft-tissue sarcomas. PATIENTS AND METHODS: This phase 1b dose-escalation trial of NC-6300 monotherapy employed a Bayesian continuous reassessment method design. NC-6300 was administered on day 1 of every 21-day cycle, with epirubicin-equivalent dose increments from 125 to 215 mg/m2. Safety, efficacy, quality of life, and pharmacokinetic profile of NC-6300 monotherapy were evaluated. RESULTS: Twenty-nine subjects (16 male) were enrolled: 17 with soft-tissue sarcoma, one with osteosarcoma, and 11 with other solid tumors. Observed dose-limiting toxicities included thrombocytopenia, stomatitis, lung infection, and febrile neutropenia. The most common grade 3/4 adverse events were neutropenia (59%), anemia (24%), thrombocytopenia (24%), and febrile neutropenia (21%). MTD and RP2D were determined to be 185 mg/m2 and 150 mg/m2, respectively. The objective response rate in the evaluable population was 11%. Partial response was observed in angiosarcoma and endometrial stromal sarcoma. A dose-dependent increase was observed in both total and released epirubicin concentrations. CONCLUSIONS: NC-6300 was well tolerated with a manageable side effect profile, despite the MTD and RP2D being higher than conventional epirubicin doses. A signal of preliminary activity was observed in angiosarcoma. NC-6300 warrants further investigation in patients with advanced solid tumors, including sarcoma.
Subject(s)
Epirubicin/analogs & derivatives , Epirubicin/administration & dosage , Proteins/administration & dosage , Sarcoma/drug therapy , Adult , Aged , Dose-Response Relationship, Drug , Epirubicin/adverse effects , Female , Humans , Male , Maximum Tolerated Dose , Middle Aged , Neoplasm Metastasis , Neutropenia/chemically induced , Neutropenia/epidemiology , Proteins/adverse effects , Sarcoma/genetics , Sarcoma/pathologyABSTRACT
BACKGROUND/AIM: Epidoxorubicin is an anthracycline agent. The present study was undertaken to compare the antileukemic potential of epidoxorubicin and its two formamidine analogs containing either a morpholine moiety (EPIFmor) or a hexamethyleneimine moiety (EPIFhex) in the amidine group. MATERIALS AND METHODS: The experiments were performed in vitro on MOLT-4 cells using spectrophotometry, Coulter electrical impedance, flow cytometry, and light microscopy methods. RESULTS: The leukemia cell responses to the action of the anthracyclines were manifested in their different viability, count and volume, degree of apoptosis and necrosis, activity of caspases -8, -9, and -3/7, mitochondrial membrane potential, and in the cell-cycle distribution. In general, epidoxorubicin appeared to be the most active, and EPIFmor was more active than EPIFhex against MOLT-4 cells. CONCLUSION: The structural modifications of epidoxorubicin in the amidine group were responsible for the varied action of its formamidine analogs on human acute lymphoblastic leukemia cells.
Subject(s)
Amidines/pharmacology , Anthracyclines/pharmacology , Antineoplastic Agents/pharmacology , Caspases/metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism , Amidines/chemistry , Anthracyclines/chemistry , Antineoplastic Agents/chemistry , Apoptosis , Cell Cycle/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Epirubicin/analogs & derivatives , Humans , In Vitro Techniques , Membrane Potentials/drug effects , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapyABSTRACT
INTRODUCTION: The use of hybrid molecules has become one of the most significant approaches in new cytotoxic drug design. This study describes synthesis and characterization of conjugates consisting of two well-known and characterized chemotherapeutic agents: methotrexate (MTX) and epirubicin (EPR). The synthesized conjugates combine two significant anticancer strategies: combinatory therapy and targeted therapy. These two drugs were chosen because they have different mechanisms of action, which can increase the anticancer effect of the obtained conjugates. MTX, which is a folic acid analog, has high cytotoxic properties and can serve as a targeting moiety that can reach folate receptors (FRs) overexpresing tumor cells. Combination of nonselective drugs such as EPR with MTX can increase the selectivity of the obtained conjugates, while maintaining the high cytotoxic properties. MATERIALS AND METHODS: Conjugates were purified by RP-HPLC and the structure was investigated by MS and MS/MS methods. The effect of the conjugates on proliferation of LoVo, LoVo/Dx, MCF-7 and MV-4-11 human cancer cell lines was determined by SRB or MTT assay. RESULTS: The conjugation reaction results in the formation of monosubstituted (α, γ) and disubstituted MTX derivatives. In vitro proliferation data demonstrate that the conjugates synthesized in our study show lower cytotoxic properties than both chemotherapeutics used alone. DISCUSSION: Epirubicin cytotoxicity was not observed in obtained conjugates. Effective drugs release after internalization needs further investigation.
Subject(s)
Drug Design , Epirubicin/analogs & derivatives , Methotrexate/analogs & derivatives , Neoplasms/drug therapy , Antineoplastic Agents/pharmacology , Epirubicin/pharmacology , Epirubicin/therapeutic use , Humans , Mass Spectrometry , Methotrexate/pharmacology , Methotrexate/therapeutic useABSTRACT
Background K-912 also known as NC-6300 is a novel epirubicin pro-drug conjugate developed using micellar nanoparticle technology. We conducted a first-in-human, Phase 1, open-label, non-randomized dose escalation study to evaluate the safety, tolerability, efficacy, and pharmacokinetics of K-912 administered as monotherapy in patients with advanced or recurrent solid tumors. Methods Patients aged 41 to 72 years with histologically or cytologically confirmed advanced or recurrent malignant solid tumors either refractory to standard therapy or had no other viable treatment options were enrolled. K-912 was administered as a 10-min intravenous infusion every three weeks. Doses were increased in a step-wise manner based on a predetermined series: 15, 30, 60, 80, 100, 130, 170, and 225 mg/m2. The appropriateness of doses above 60 mg/m2 was assessed using a Bayesian continual reassessment model. Treatment-emergent adverse events and tumor response were evaluated according to internationally accepted criteria. Results Nineteen patients were treated with K-912. No additional adverse events expected with anthracyclines were observed. While the number of patients treated at the maximum tolerated dose (MTD) and the recommended phase 2 dose (RP2D) were small, MTD and RP2D were established to be 170 mg/m2. Partial response was observed in one patient with breast cancer treated at 100 mg/m2, yielding an objective response rate of 5% (1/19). Stable disease was observed in 10 patients. The human pharmacokinetic profile of K-912 was consistent with that observed from nonclinical studies in rats and monkeys. Conclusions This study showed that K-912 was well tolerated in patients with various solid tumors and exhibited less toxicity than conventional epirubicin formulations.
Subject(s)
Epirubicin/analogs & derivatives , Micelles , Neoplasm Recurrence, Local/drug therapy , Neoplasms/drug therapy , Proteins/administration & dosage , Adult , Aged , Epirubicin/administration & dosage , Epirubicin/adverse effects , Epirubicin/pharmacokinetics , Epirubicin/therapeutic use , Female , Humans , Leukopenia/chemically induced , Male , Maximum Tolerated Dose , Middle Aged , Neoplasm Recurrence, Local/metabolism , Neoplasms/metabolism , Polymers/administration & dosage , Polymers/adverse effects , Polymers/pharmacokinetics , Polymers/therapeutic use , Proteins/adverse effects , Proteins/pharmacokinetics , Proteins/therapeutic use , Retinal Hemorrhage/chemically induced , Ventricular Function, Left/drug effectsABSTRACT
Epirubicin (EPI) is one of the most widely used anticarcinogens; however, serious side effects, including cardiomyopathy and congestive heart failure, limit its longterm administration. To overcome this problem, the HAIYPRH peptide ligand was used with EPI in the synthesis of a HAIYPRHEPI conjugate. The anticancer activity and cellular uptake of the conjugate were measured and evaluated. The results of the present study indicated that the cytotoxicity of HAIYPRHEPI was correlated with the expression of the cell surface transferrin receptor (TfR). The conjugate exerted high cytotoxicity and proapoptotic function when in an LN229 glioma cell line, which overexpresses surface TfR. It was hypothesized that transferrin (Tf) can promote cytotoxicity. Conversely, the conjugate exhibited very low cytotoxicity and proapoptotic function in a U87 glioma cell line, in which surface TfR expression was undetectable. In addition, fluorescence microscopy and flow cytometry methods were used to evaluate cellular uptake, and the results of these methods were consistent with the present hypotheses. The conjugate cellular uptake of the conjugate in LN229 cells was markedly higher compared with that in U87 cells, and it was hypothesized that Tf can enhance the uptake in LN229 cells. The cytotoxicity of HAIYPRHEPI was dependent upon the expression of surface TfR. Considering that the majority of cancer cells have high rates of iron uptake and surface TfR is generally overexpressed on cancer cells, it was speculated by the authors that HAIYPRHEPI may form part of an effective strategy for increasing the selectivity of EPI for cancer cells, as well as reducing its systemic toxicity. To confirm the hypothesis, the effects of HAIYPRHEPI on noncancerous cell lines were investigated. A future study will examine the side effects of HAIYPRHEPI, using a suitable delivery system in an animal model.
Subject(s)
Antigens, CD/metabolism , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Epirubicin/analogs & derivatives , Epirubicin/pharmacology , Peptides/chemistry , Peptides/pharmacology , Receptors, Transferrin/metabolism , Amino Acid Sequence , Antineoplastic Agents/pharmacokinetics , Cell Line , Cell Line, Tumor , Cell Survival/drug effects , Epirubicin/pharmacokinetics , Glioma/drug therapy , Glioma/metabolism , Humans , Peptides/pharmacokineticsABSTRACT
Non-Hodgkin lymphoma is a common childhood T-cell and B-cell neoplasm that originates primarily from lymphoid tissue. Cutaneous involvement can be in the form of a primary extranodal lymphoma, or secondary to metastasis from a non-cutaneous location. The latter is uncommon, and isolated cutaneous involvement is rarely reported. We report a case of isolated secondary cutaneous involvement from nodal anaplastic large cell lymphoma (CD30 + and ALK +) in a 7-year-old boy who was on chemotherapy. This case is reported for its unusual clinical presentation as an acute febrile, generalized papulonodular eruption that mimicked deep fungal infection, with the absence of other foci of systemic metastasis.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Lymphoma, Large-Cell, Anaplastic/drug therapy , Lymphoma, Large-Cell, Anaplastic/pathology , Skin Neoplasms/drug therapy , Skin Neoplasms/secondary , Child , Cyclophosphamide/administration & dosage , Epirubicin/administration & dosage , Epirubicin/analogs & derivatives , Follow-Up Studies , Humans , Male , Neoplasm Invasiveness , Neoplasm Staging , Prednisolone/administration & dosage , Rare Diseases , Risk Assessment , Skin Neoplasms/pathology , Treatment Outcome , Vincristine/administration & dosageABSTRACT
The objectives of the present study were to develop functional targeting epirubicin liposomes for transferring drugs across the blood-brain barrier (BBB), treating glioblastoma, and disabling neovascularization. The studies were performed on glioblastoma cells in vitro and on glioblastoma-bearing mice. The results showed that the constructed liposomes had a high encapsulation efficiency for drugs (>95%), suitable particle size (109 nm), and less leakage in the blood component-containing system; were significantly able to be transported across the BBB; and exhibited efficacies in killing glioblastoma cells and in destroying glioblastoma neovasculature in vitro and in glioblastoma-bearing mice. The action mechanisms of functional targeting epirubicin liposomes correlated with the following features: the long circulation in the blood system, the ability to be transported across the BBB via glucose transporter-1, and the targeting effects on glioblastoma cells and on the endothelial cells of the glioblastoma neovasculature via the integrin ß3 receptor. In conclusion, functional targeting epirubicin liposomes could be used as a potential therapy for treating brain glioblastoma and disabling neovascularization in brain glioblastomas.
Subject(s)
Angiogenesis Inhibitors/administration & dosage , Antibiotics, Antineoplastic/administration & dosage , Brain Neoplasms/drug therapy , Epirubicin/administration & dosage , Glioblastoma/drug therapy , Glucosides/metabolism , Nanoparticles , Peptides, Cyclic/metabolism , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/metabolism , Animals , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/metabolism , Blood-Brain Barrier/metabolism , Brain Neoplasms/blood supply , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Capillary Permeability , Cell Line, Tumor , Chemistry, Pharmaceutical , Dose-Response Relationship, Drug , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Endothelial Cells/pathology , Epirubicin/analogs & derivatives , Epirubicin/chemistry , Epirubicin/metabolism , Glioblastoma/blood supply , Glioblastoma/metabolism , Glioblastoma/pathology , Glucosides/chemistry , Humans , Liposomes , Male , Mice, Inbred BALB C , Mice, Nude , Neovascularization, Pathologic , Peptides, Cyclic/chemistry , Spheroids, Cellular , Time Factors , Tissue Distribution , Xenograft Model Antitumor AssaysABSTRACT
Anthracycline antibiotics are extensively applied to clinical antitumor therapy. The binding mode and mechanism of a new anthracycline 3'-azido-epirubicin (AEPI) with calf thymus deoxyribonucleic acid (ctDNA) were investigated employing multiple spectroscopy techniques in Tris-HCl buffer solution (pH 7.4). Effect of pH on the interaction was provided to determine the proper environment for whole research. Iodide quenching studies and fluorescence polarization measurement indicated that ctDNA quenched the fluorescence of AEPI significantly via intercalation binding mode. The binding constants and binding sites for the interaction were calculated. From binding constant dependence on the temperature, static quenching mechanism of AEPI by ctDNA was confirmed based on the Stern-Volmer equation. Additionally, the thermodynamic parameters for the reaction revealed that the van der Waals force and hydrogen bonding were the main acting forces in the binding process. Molecular modeling result indicated that the hydrogen bonding played a major role in the binding of AEPI to ctDNA.
Subject(s)
Anthracyclines/chemistry , Azides/chemistry , DNA/chemistry , Epirubicin/analogs & derivatives , Epirubicin/chemistry , Intercalating Agents/chemistry , Animals , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/metabolism , Azides/metabolism , Binding Sites , Cattle , DNA/metabolism , Epirubicin/metabolism , Hydrogen Bonding , Intercalating Agents/metabolism , Kinetics , Models, Molecular , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet , Temperature , ThermodynamicsABSTRACT
It is expected that the incidence of various adverse effects of anticancer agents maybe decreased owing to the reduced drug distribution in normal tissue. Anticancer agent incorporating nanoparticles including micelles and liposomes can evade non-specific capture by the reticuloendothelial system because the outer shell of the nanoparticles is covered with polyethylene glycol. Consequently, the micellar and liposomal carrier can be delivered selectively to a tumor by utilizing the enhanced permeability and retention effect. Presently, several anticancer agent-incorporating nano-carrier systems are under preclinical and clinical evaluation. Several drug delivery system formulations have been approved worldwide. Regarding a pipeline of clinical development of anticancer agent incorporating micelle carrier system, several clinical trials are now underway not only in Japan but also in other countries. A Phase 3 trial of NK105, a paclitaxel incorporating micelle is now underway. In this paper, preclinical and clinical studies of NK105, NC-6004, cisplatin incorporating micelle, NC-6300, epirubicin incorporating micelle and the concept of cancer stromal targeting therapy using nanoparticles and monoclonal antibodies against cancer related stromal components are reviewed.
Subject(s)
Antineoplastic Agents/administration & dosage , Drug Delivery Systems , Micelles , Nanomedicine/trends , Nanoparticles/administration & dosage , Neoplasms/drug therapy , Neoplasms/metabolism , Animals , Antineoplastic Agents/pharmacology , Chromatography, High Pressure Liquid , Cisplatin/administration & dosage , Clinical Trials, Phase I as Topic , Drug Discovery , Epirubicin/administration & dosage , Epirubicin/analogs & derivatives , Gas Chromatography-Mass Spectrometry , Humans , Mass Spectrometry , Neoplasms, Experimental/drug therapy , Neoplasms, Experimental/metabolism , Organoplatinum Compounds/administration & dosage , Paclitaxel/administration & dosage , Paclitaxel/analogs & derivatives , Polyethylene Glycols/administration & dosage , Polyglutamic Acid/administration & dosage , Polyglutamic Acid/analogs & derivatives , Proteins/administration & dosageABSTRACT
Epirubicin (EPI) is a broad spectrum antineoplastic drug, commonly used as a chemotherapy method to treat osteosarcoma. However, its application has been limited by many side-effects. Therefore, targeted drug delivery to bone has been the aim of current anti-bone-tumor drug studies. Due to the exceptional affinity of Bisphosphonates (BP) to bone, 1-amino-ethylene-1, 1-dephosphate acid (AEDP) was chosen as the bone targeting moiety for water-soluble macromolecular drug delivery systems of oxidized-dextran (OXD) to transport EPI to bone in this article. The bone targeting drug of AEDP-OXD-EPI was designed for the treatment of malignant bone tumors. The successful conjugation of AEDP-OXD-EPI was confirmed by analysis of FTIR and (1)H-NMR spectra. To study the bone-seeking potential of AEDP-OXD-EPI, an in vitro hydroxyapatite (HAp) binding assay and an in vivo experiment of bone-targeting capacity were established. The effectiveness of AEDP-OXD-EPI was demonstrated by inducing apoptosis and necrosis of MG-63 tumor cell line. The obtained experimental data indicated that AEDP-OXD-EPI is an ideal bone-targeting anti-tumor drug.
Subject(s)
Antibiotics, Antineoplastic/chemical synthesis , Bone and Bones/drug effects , Dextrans/chemistry , Diphosphonates/chemistry , Drug Carriers/chemical synthesis , Epirubicin/analogs & derivatives , Epirubicin/chemical synthesis , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/pharmacology , Antibiotics, Antineoplastic/therapeutic use , Apoptosis/drug effects , Bone Neoplasms/drug therapy , Bone Neoplasms/metabolism , Bone Neoplasms/pathology , Bone and Bones/metabolism , Cell Line, Tumor , Dextrans/pharmacology , Diphosphonates/pharmacology , Drug Carriers/chemistry , Drug Carriers/pharmacology , Epirubicin/chemistry , Epirubicin/pharmacology , Epirubicin/therapeutic use , Femur/drug effects , Femur/metabolism , Humans , Molecular Structure , Osteosarcoma/drug therapy , Osteosarcoma/metabolism , Osteosarcoma/pathology , Solubility , Water/chemistryABSTRACT
Epirubicin is widely used to treat various human tumors. However, it is difficult to achieve a sufficient antitumor effect because of dosage limitation to prevent cardiotoxicity. We hypothesized that epirubicin-incorporating micelle would reduce cardiotoxicity and improve the antitumor effect. NC-6300 comprises epirubicin covalently bound to PEG polyaspartate block copolymer through an acid-labile hydrazone bond. The conjugate forms a micellar structure of 40-80 nm in diameter in an aqueous milieu. NC-6300 (10, 15 mg/kg) and epirubicin (10 mg/kg) were given i.v. three times to mice bearing s.c. or liver xenograft of human hepatocellular carcinoma Hep3B cells. Cardiotoxicity was evaluated by echocardiography in C57BL/6 mice that were given NC-6300 (10 mg/kg) or epirubicin (10 mg/kg) in nine doses over 12 weeks. NC-6300 showed a significantly potent antitumor effect against Hep3B s.c. tumors compared with epirubicin. Moreover, NC-6300 also produced a significantly longer survival rate than epirubicin against the liver orthotopic tumor of Hep3B. With respect to cardiotoxicity, epirubicin-treated mice showed significant deteriorations in fractional shortening and ejection fraction. In contrast, cardiac functions of NC-6300 treated mice were no less well maintained than in control mice. This study warrants a clinical evaluation of NC-6300 in patients with hepatocellular carcinoma or other cancers.
Subject(s)
Antibiotics, Antineoplastic/adverse effects , Antibiotics, Antineoplastic/pharmacology , Antineoplastic Agents/pharmacology , Epirubicin/analogs & derivatives , Heart/drug effects , Micelles , Proteins/adverse effects , Proteins/pharmacology , Animals , Antineoplastic Agents/adverse effects , Carcinoma, Hepatocellular/drug therapy , Cell Line, Tumor , Epirubicin/adverse effects , Epirubicin/pharmacology , Female , Humans , Liver Neoplasms/drug therapy , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Nude , Random Allocation , Tissue Distribution/drug effects , Xenograft Model Antitumor AssaysABSTRACT
Epirubicin was developed as a semi-synthetic anthracycline derivative to circumvent the cardiotoxic limitations associated with the use of doxorubicin in the clinic. Anthracycline compounds have been demonstrated to form covalent drug-DNA adducts utilising endogenous and exogenous sources of formaldehyde; however, previous investigations of the formation of epirubicin-DNA adducts provide conflicting evidence for adduct formation. This work provides evidence that epirubicin acts to form drug-DNA adducts at physiologically relevant concentrations and demonstrates that the rate of formation of epirubicin-DNA adducts is slower than that observed for other anthracycline compounds, explaining why they are only detectable under defined experimental conditions. Formation of covalent epirubicin-DNA adducts improves the apoptotic profile of epirubicin and provides opportunities to overcome drug resistance and cardiotoxic limitations.
Subject(s)
Anthracyclines/metabolism , Antibiotics, Antineoplastic/pharmacology , Cytotoxins/metabolism , DNA Adducts/drug effects , Epirubicin/analogs & derivatives , Epirubicin/pharmacology , Apoptosis/drug effects , Blotting, Western , Carbohydrates/chemistry , Cell Line, Tumor , Coloring Agents , Cross-Linking Reagents , Female , Formaldehyde/chemistry , Humans , Hydroxylation , Propidium , Rhodamines , Transcription, Genetic/drug effectsABSTRACT
Cyclin-dependent kinase 10 (Cdk10) is a Cdc2-related kinase and plays an essential role in the progression from the G2 to M phase of the cell cycle. However, relative little is known about its expression pattern, clinical relevance, and biological function in hepatocellular carcinoma (HCC). In the present study, we investigated the mRNA and protein expression levels of Cdk10 in 127 pairs of HCC samples and adjacent nontumorous liver tissues and evaluated its clinical significance. Additionally, we assessed the effects of restoration of Cdk10 on cell proliferation and drug sensitivity in HCC cells. We showed that the Cdk10 mRNA and protein expression was markedly decreased in HCC samples compared to adjacent nontumorous liver tissues. Quantitative real-time polymerase chain reaction and immunohistochemical studies revealed that reduced Cdk10 expression was significantly associated with alpha-fetoprotein levels, tumor size, and tumor stage. Ectopic expression of Cdk10 reduced HCC cell proliferation, blocked the cell cycle at the G0-G1 phase, as well as inhibited cell migration and anchorage-independent growth. Additionally, Cdk10 overexpression enhanced the chemosensitivity of HCC cells to cisplatin and epidoxorubicin, two chemotherapeutic agents commonly used in HCC. These data collectively demonstrate that reduced Cdk10 expression is closely linked to HCC development and progression. Restoration of its expression may have therapeutic benefits in treating this malignancy.
Subject(s)
Carcinoma, Hepatocellular/enzymology , Carcinoma, Hepatocellular/genetics , Cyclin-Dependent Kinases/genetics , Cyclin-Dependent Kinases/metabolism , Liver Neoplasms/enzymology , Liver Neoplasms/genetics , Aged , Base Sequence , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Cisplatin/pharmacology , Down-Regulation , Drug Resistance, Neoplasm , Epirubicin/analogs & derivatives , Epirubicin/pharmacology , Female , Glucuronates/pharmacology , Humans , Immunohistochemistry , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Male , Middle Aged , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Neoplasm/genetics , RNA, Neoplasm/metabolismABSTRACT
The C(3)-monoamine on the carbohydrate moiety (daunosamine -NH(2)-3') of epirubicin was reacted under anhydrous conditions with succinimidyl 4,4-azipentanoate to create a covalent UV-photoactivated epirubicin-(C(3)-amide) intermediate with primary amine-reactive properties. A synthetic covalent bond between the UV-photoactivated epirubicin-(C(3)-amide) intermediate and the É-amine of lysine residues within the amino acid sequence of anti-HER2/neu monoclonal immunoglobulin was subsequently created by exposure to UV light (354 nm) for 15 minutes. Size-separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis combined with immunodetection analysis and chemiluminescent autoradiographic imaging revealed a lack of IgG-IgG polymerization or degradative protein fragmentation of the covalent epirubicin-(C(3)-amide)-[anti-HER2/neu] immunochemotherapeutic. Retained binding-avidity of epirubicin-(C(3)-amide)-[anti-HER2/neu] was validated by cell-ELISA utilizing monolayer populations of chemotherapeutic-resistant mammary adenocarcinoma SKBr-3 which highly overexpress membrane-associated HER2/neu complexes. Between epirubicin-equivalent concentrations of 10(-10) to 10(-6) M the covalent epirubicin-(C(3)-amide)-[anti-HER2/neu] immunochemotherapeutic consistently evoked levels of cytotoxic anti-neoplastic potency that were highly analogous to chemotherapeutic-equivalent concentrations of epirubicin. Cytotoxic anti-neoplastic potency of epirubicin-(C(3)-amide)-[anti-HER2/neu] against chemotherapeutic-resistant mammary adenocarcinoma SKBr-3 challenged with epirubicin-(C(3)-amide)-[anti-HER2/neu] at an epirubicin-equivalent concentration of 10(-6) M was 88.5% (e.g., 11.5% residual survival). Between final epirubicin-equivalent concentrations of 10(-8) and 10(-7) M there was a marked threshold increase in the mean cytotoxic anti-neoplastic activity for epirubicin-(C(3)-amide)-[anti-HER2/neu] from 9.9% to 66.9% (90.2% to 33.1% residual survival).
Subject(s)
Antibiotics, Antineoplastic/chemical synthesis , Breast Neoplasms/drug therapy , Epirubicin/analogs & derivatives , Receptor, ErbB-2/antagonists & inhibitors , Adenocarcinoma/drug therapy , Adenocarcinoma/enzymology , Adenocarcinoma/genetics , Adenocarcinoma/pathology , Antibiotics, Antineoplastic/chemistry , Antibiotics, Antineoplastic/pharmacology , Breast Neoplasms/enzymology , Breast Neoplasms/immunology , Breast Neoplasms/pathology , Cell Line, Tumor , Enzyme-Linked Immunosorbent Assay , Epirubicin/chemical synthesis , Epirubicin/pharmacology , Female , Humans , Immunoglobulin G/immunology , Photochemical Processes , Tumor Cells, Cultured , Ultraviolet RaysABSTRACT
BACKGROUND: The most appropriate timing of chemotherapy and hormone therapy administration is a critical issue in early breast cancer patients. The purpose of our study was to compare the efficacy of concurrent vs sequential administration of adjuvant chemotherapy and tamoxifen. METHODS: Women with node-positive primary breast cancer were randomly assigned to receive tamoxifen (20 mg/d for 5 years) during (concurrent arm) or after (sequential arm) adjuvant chemotherapy. Chemotherapy consisted of alternating regimens of cyclophosphamide, epidoxorubicin, and 5-fluorouracil and cyclophosphamide, methotrexate, and 5-fluorouracil every 21 days for a total of 12 cycles. The primary endpoint was overall survival (OS), and secondary endpoints were toxic effects and disease-free survival (DFS). No provision for interim analyses was made in the original study protocol. Survival curves were estimated by the Kaplan-Meier method. Multivariable Cox regression models, adjusted for age, menopausal status, tumor stage, and lymph node and hormone receptor status, were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: From 1985 to 1992, 431 patients were randomly assigned and studied according to the intention-to-treat principle. After a maximum of 15.4 years of follow-up (median 12.3 years), the estimated actuarial 10-year OS was equivalent for the two study arms (concurrent arm: 111 patients, 66%, 95% CI = 59% to 72%; sequential arm: 114 patients, 65%, 95% CI = 59% to 72%, P = .86). No differences in DFS and toxic effects were evident. Four interim analyses were performed, but no alpha error adjustment was necessary because of the largely negative results of this final analysis (sequential vs concurrent arm: HR of death = 1.06, 95% CI = 0.78 to 1.44, P = .76; HR of relapse = 1.16, 95% CI = 0.88 to 1.52, P = .36). CONCLUSIONS: No statistically significant differences in OS, DFS, and toxic effects between concurrent and sequential adjuvant chemo- and hormone therapies were observed. Our study does not support the superiority of one schedule of chemo- and hormone-therapy administration over the other. However, because of the limited statistical power of the study, these results must be considered with caution.
Subject(s)
Antineoplastic Agents, Hormonal/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Breast Neoplasms/drug therapy , Adult , Axilla , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Breast Neoplasms/surgery , Chemotherapy, Adjuvant , Confounding Factors, Epidemiologic , Cyclophosphamide/administration & dosage , Disease-Free Survival , Drug Administration Schedule , Epirubicin/administration & dosage , Epirubicin/analogs & derivatives , Female , Fluorouracil/administration & dosage , Follow-Up Studies , Glucuronates/administration & dosage , Humans , Italy , Kaplan-Meier Estimate , Lymph Node Excision , Mastectomy, Segmental , Methotrexate/administration & dosage , Middle Aged , Multivariate Analysis , Neoplasm Staging , Neoplasms, Hormone-Dependent/drug therapy , Prognosis , Proportional Hazards Models , Receptors, Estrogen/metabolism , Receptors, Progesterone/metabolism , Risk Factors , Tamoxifen/administration & dosage , Treatment OutcomeABSTRACT
Anthracyclines have long been considered to be among the most active agents clinically available for the treatment of breast cancer despite their toxicity. To improve their pharmacological profiles, a new macromolecular prodrug, denoted NC-6300, was synthesized. NC-6300 comprises epirubicin covalently bound to polyethylene-glycol polyaspartate block copolymer through an acid-labile hydrazone bond. The conjugate forms a micellar structure spontaneously in aqueous media with a diameter of 60-70 nm. The block copolymers are partially substituted with hydrophobic benzyl groups to stabilize the micellar structure. The present study was designed to confirm that polymeric micelles incorporating epirubicin through an acid-labile linker improve the therapeutic index and achieve a broad range of therapeutic doses. Pharmacokinetic studies in rats showed highly enhanced plasma retention of NC-6300 compared with native epirubicin. The maximal tolerated doses in mice of NC-6300 and native epirubicin were 25 and 9 mg/kg, respectively, when administered three times with a 4-day interval between each dose. NC-6300 at 15 and 20 mg/kg with the same administration schedule regressed a Hep3B human hepatic tumor with slight and transient bodyweight loss. Remarkably, NC-6300 also inhibited growth of an MDA-MB-231 human breast tumor at the same dosage. In contrast, native epirubicin at 7 mg/kg administered three times with a 4-day interval was only able to slow tumor growth. Tissue distribution studies of NC-6300 showed efficient free epirubicin released in the tumor at 74% by area under the concentration-time curve (AUC) evaluation, supporting the effectiveness of NC-6300. In conclusion, NC-6300 improved the potency of epirubicin, demonstrating the advantage of NC-6300 attributable to the efficient drug release in the tumor.
Subject(s)
Anthracyclines/pharmacology , Antineoplastic Agents/pharmacology , Epirubicin/analogs & derivatives , Proteins/pharmacology , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacokinetics , Cell Line, Tumor , Chemistry, Pharmaceutical , Drug Stability , Epirubicin/chemistry , Epirubicin/pharmacokinetics , Epirubicin/pharmacology , Epirubicin/toxicity , Female , Humans , Male , Mice , Mice, Inbred BALB C , Micelles , Proteins/chemistry , Proteins/pharmacokinetics , Proteins/toxicity , Rats , Rats, Wistar , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: Epithelial ovarian cancer accounts for about 90% of all cases of ovarian cancer. Debulking surgery and six courses of platinum-based chemotherapy results in complete clinical remission (CCR) in up to 75% of cases. However, 75% of the responders will relapse within a median time of 18 to 28 months and only 20% to 40% of women will survive beyond five years. It has been suggested that maintenance chemotherapy could assist in prolonging remission. To date, there has not been a systematic review on the impact of maintenance chemotherapy for epithelial ovarian cancer. OBJECTIVES: To assess the effectiveness and toxicity of maintenance chemotherapy for epithelial ovarian cancer and to evaluate the impact on quality of life (QoL). SEARCH STRATEGY: We searched the Cochrane Gynaecological Cancer Review Group Specialized Register, The Cochrane Central Register of Controlled Trails (CENTRAL, The Cochrane Library Issue1, 2009), MEDLINE, EMBASE, PubMed, CBMdisc, CNKI and VIP (to May 2009). We collected information from ongoing trials, checked reference lists of published articles and consulted experts in the field. SELECTION CRITERIA: Randomised controlled trials (RCTs) comparing maintenance chemotherapy with no further intervention, maintenance radiotherapy or other maintenance therapy. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed trials for eligibility and quality and extracted data. We analysed overall survival (OS) and progression free survival (PFS) rates as dichotomous variables. Toxicity and QoL data were extracted where present. All analyses were based on intention to treat (ITT) on the endpoint of survival. We also analysed data by subgroups of drugs. MAIN RESULTS: We included six trials(902 women). When all chemotherapy regimens were combined, meta-analysis indicated no significant difference in 3, 5 and 10-year OS or PFS. For 5-year OS, the combined relative risk (RR) was 1.07 (95% confidence interval (CI) 0.91 to 1.27) and for the 5-year PFS the combined RR was 1.18 (95% CI 0.88 to 1.58). Results were very similar when trials of different regimens were analysed. Comparing chemotherapy with radiotherapy, only the RR for 10-year PFS in pathological complete remission was in favour of whole abdominal radiotherapy 0.51 (95% CI 0.27 to 1.00), while 3 and 5-year OS rates have no significant difference between the two groups. AUTHORS' CONCLUSIONS: There is no evidence to suggest that the use of platinum agents or doxorubicin used as maintenance chemotherapy is more effective than observation alone. Further investigations regarding the effect of paclitaxel used as maintenance chemotherapy are required.
Subject(s)
Antineoplastic Agents/therapeutic use , Ovarian Neoplasms/drug therapy , Cisplatin/administration & dosage , Cyclophosphamide/administration & dosage , Disease-Free Survival , Doxorubicin/administration & dosage , Epirubicin/administration & dosage , Epirubicin/analogs & derivatives , Female , Glucuronates/administration & dosage , Humans , Ovarian Neoplasms/mortality , Ovarian Neoplasms/radiotherapy , Quality of Life , Randomized Controlled Trials as Topic , Remission Induction/methodsABSTRACT
4'-Epiadriamycin is a better-tolerated anthracycline drug, due to lesser cardiotoxicity. We report here a study of the 2:1 complex of 4'-epiadriamycin-d-(CGATCG)(2) by proton Nuclear Magnetic Resonance Spectroscopy which show the absence of sequential connectivities between C1pG2 and C5pG6 base pair steps and presence of intermolecular cross peaks of the drug and DNA. Our studies establish the role of 9OH, NH3+, 7O, 4OCH(3) groups in binding to DNA. Time-resolved fluorescence measurement and diffusion ordered spectroscopic studies reveal the formation of complex. The nonspecific interactions as well as those essential for biological activity are discussed along with its medicinal importance.