ABSTRACT
Lysolipid-containing thermosensitive liposomes (LTSL) are clinically-relevant drug nanocarriers which have been used to deliver small molecule cytostatics to tumors in combination with local hyperthermia (42⯰C) to trigger local drug release. The objective of this study was to investigate the feasibility of LTSL for encapsulation and triggered release of macromolecular drugs such as plant-derived cytotoxins. As therapeutic protein we used Mistletoe lectin-1 (ML1) - a ribosome-inactivating protein with potent cytotoxic activity in tumor cells. Model macromolecules (dextrans, albumin) and ML1 were encapsulated in small unilamellar LTSL with varying lipid compositions by the thin film hydration method and extrusion. LTSLs showed molecular weight dependent heat-triggered release of the loaded cargo. The most promising composition, ML1 formulated in LTSL composed of 86:10:4 %mol DPPC:MSPC:DSPE-PEG2000, was further studied for bioactivity against murine CT26 colon carcinoma cells. Confocal live-cell imaging showed uptake of released ML1 after mild hyperthermia at 42⯰C, subsequently leading to potent cytotoxicity by LTSL-ML1. Our study shows that LTSL in combination with localized hyperthermia hold promise as local tumor delivery strategy for macromolecular cytotoxins.
Subject(s)
Antineoplastic Agents, Phytogenic/administration & dosage , Colonic Neoplasms/drug therapy , Lipids/chemistry , Ribosome Inactivating Proteins, Type 2/administration & dosage , Toxins, Biological/administration & dosage , Albumins/chemistry , Animals , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Cell Line, Tumor , Chemistry, Pharmaceutical/methods , Colonic Neoplasms/pathology , Dextrans/chemistry , Drug Delivery Systems , Drug Liberation , Hot Temperature , Liposomes , Mice , Molecular Weight , Ribosome Inactivating Proteins, Type 2/chemistry , Ribosome Inactivating Proteins, Type 2/pharmacology , Temperature , Toxins, Biological/chemistry , Toxins, Biological/pharmacologyABSTRACT
Viscumin (mistletoe lectin I, MLI) in concentrations of 10-11-10-7 M causes endoplasmic reticulum stress and triggers unfolded protein response, a modulator of antitumor immunity, in target cells.
Subject(s)
Endoplasmic Reticulum Stress , Ribosome Inactivating Proteins, Type 2/pharmacokinetics , Toxins, Biological/pharmacokinetics , Animals , Caco-2 Cells , Cell Line, Tumor , Endoplasmic Reticulum Stress/drug effects , Humans , Injections, Subcutaneous , Male , Mice , Plant Lectins/administration & dosage , Plant Lectins/pharmacokinetics , Plant Lectins/pharmacology , Ribosome Inactivating Proteins, Type 2/administration & dosage , Ribosome Inactivating Proteins, Type 2/pharmacology , Toxins, Biological/administration & dosage , Toxins, Biological/pharmacology , Unfolded Protein Response/drug effectsABSTRACT
Some patients use complementary medicine. We present a patient with Hodgkin's lymphoma, scanned with 18F-FDG PET/CT for evaluation of response after chemotherapy, who was self-administering mistletoe as a homeopathic medicine product. The careful review of the images of the entire scan and patient collaboration in anamnesis were crucial to avoid a false positive result. A review of the published scientific data on the effects of mistletoe is also presented.
Subject(s)
Antineoplastic Agents, Phytogenic/adverse effects , Fluorine Radioisotopes/analysis , Fluorodeoxyglucose F18/analysis , Hodgkin Disease/diagnostic imaging , Lymph Nodes/diagnostic imaging , Materia Medica/adverse effects , Phytotherapy/adverse effects , Positron Emission Tomography Computed Tomography , Radiopharmaceuticals/analysis , Ribosome Inactivating Proteins, Type 2/adverse effects , Toxins, Biological/adverse effects , Viscum album/adverse effects , Adult , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/therapeutic use , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Bleomycin/administration & dosage , Dacarbazine/administration & dosage , Doxorubicin/administration & dosage , Female , Hodgkin Disease/drug therapy , Humans , Injections, Subcutaneous , Lymph Nodes/drug effects , Neoplasm Staging , Ribosome Inactivating Proteins, Type 2/administration & dosage , Ribosome Inactivating Proteins, Type 2/therapeutic use , Self Medication , Subcutaneous Fat/diagnostic imaging , Subcutaneous Fat/metabolism , Toxins, Biological/administration & dosage , Toxins, Biological/therapeutic use , Vinblastine/administration & dosageABSTRACT
An essential in vivo drug delivery system of a momordica anti-HIV protein, MAP30, was developed through encapsulating in chemically synthesized matrices of zirconium egg- and soy-phosphatidylcholines, abbreviated to Zr/EPC and Zr/SPC, respectively. Matrices were characterized by transmission electron microscopy and powder X-ray diffractometry studies. Zr/EPC granule at an approximate diameter of 69.43±7.78 nm was a less efficient encapsulator than the granule of Zr/SPC. Interlayer spacing of the matrices encapsulating MAP30 increased from 8.8 and 9.7 Å to 7.4 and 7.9 nm, respectively. In vivo kinetics on degradation and protein release was performed by analyzing the serum sampling of intravenously injected SPF chickens. The first order and biphasic variations were obtained for in vivo kinetics using equilibrium dialysis. Antimicrobial and anti-HIV assays yielded greatly decreased MIC50 and EC50 values of nanoformulated MAP30. An acute toxicity of MAP30 encapsulated in Zr/EPC occurred at a single intravenous dose above 14.24 mg/kg bw in NIH/KM/ICR mice. The folding of MAP30 from Zr/EPC sustained in vivo chickens for more than 8 days in high performance liquid chromatography assays. These matrices could protect MAP30 efficiently with strong structure retention, lowered toxicity and prolonged in vivo life.
Subject(s)
Anti-Bacterial Agents/pharmacology , Anti-HIV Agents/pharmacology , Bacteria/drug effects , Drug Delivery Systems , HIV-1/drug effects , Nanocapsules/chemistry , Phosphatidylcholines/chemistry , Ribosome Inactivating Proteins, Type 2/metabolism , Zirconium/chemistry , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/metabolism , Anti-HIV Agents/chemistry , Anti-HIV Agents/metabolism , Injections, Intravenous , Mice , Mice, Inbred Strains , Molecular Structure , Nanocapsules/administration & dosage , Particle Size , Phosphatidylcholines/administration & dosage , Ribosome Inactivating Proteins, Type 2/administration & dosage , Ribosome Inactivating Proteins, Type 2/chemistry , Surface Properties , Zirconium/administration & dosageABSTRACT
Phase I oncology clinical trials are designed to identify the optimal dose that will be recommended for phase II trials. This dose is typically defined as the dose associated with a certain probability of severe toxicity during the first cycle of treatment, although toxicity is repeatedly measured over cycles on an ordinal scale. We propose a new adaptive dose-finding design using longitudinal measurements of ordinal toxic adverse events, with proportional odds mixed-effect models. Likelihood-based inference is implemented. The optimal dose is then the dose producing a target rate of severe toxicity per cycle. This model can also be used to identify cumulative or late toxicities. The performances of this approach were compared with those of the continual reassessment method in a simulation study. Operating characteristics were evaluated in terms of correct identification of the target dose, distribution of the doses allocated and power to detect trends in the risk of toxicities over time. This approach was also used to reanalyse data from a phase I oncology trial. Use of a proportional odds mixed-effect model appears to be feasible in phase I dose-finding trials, increases the ability of selecting the correct dose and provides a tool to detect cumulative effects.
Subject(s)
Antineoplastic Agents/administration & dosage , Clinical Trials, Phase I as Topic/methods , Likelihood Functions , Longitudinal Studies , Maximum Tolerated Dose , Models, Statistical , Adult , Antineoplastic Agents/toxicity , Computer Simulation , Humans , Neoplasms/drug therapy , Plant Preparations/administration & dosage , Plant Preparations/adverse effects , Ribosome Inactivating Proteins, Type 2/administration & dosage , Ribosome Inactivating Proteins, Type 2/adverse effects , Toxins, Biological/administration & dosage , Toxins, Biological/adverse effectsABSTRACT
BACKGROUND: Ribosome-inactivating proteins (RIP) have been studied in the search for toxins that could be used as immunotoxins for cancer treatment. Pulchellin, a type 2 RIP, is suggested to induce immune responses that have a role in controlling cancer. METHODS: The percentage of dendritic cells and CD4(+) and CD8(+) T cells in the spleen (flow cytometry), cytokines' release by PECs and splenocytes (ELISA) and nitric oxide production by PECs (Griess assay) were determined from tumor-bearing mice injected intratumorally with 0.1 ml of pulchellin at 0.75 µg/kg of body weight. Statistical analysis was performed by one-way ANOVA with Tukey's post hoc test. RESULTS: Pulchellin-treated mice showed significant immune system activation, characterized by increased release of IFN-γ and Th2 cytokines (IL-4 and IL-10), while IL-6 and TGF-ß levels were decreased. There was also an increase in macrophage's activation, as denoted by the higher percentage of macrophages expressing adhesion and costimulatory molecules (CD54 and CD80, respectively). CONCLUSIONS: Our results suggest that pulchellin is promising as an adjuvant in breast cancer treatment.
Subject(s)
Abrus/chemistry , Breast Neoplasms/drug therapy , Breast Neoplasms/immunology , Plant Proteins/administration & dosage , Ribosome Inactivating Proteins, Type 2/administration & dosage , Animals , CD8-Positive T-Lymphocytes/immunology , Cell Line, Tumor , Cytokines/immunology , Female , Humans , Macrophages/drug effects , Mice , Mice, Inbred BALB C , Spleen/immunology , Th2 Cells/immunologyABSTRACT
Safety of aviscumine by subcutaneous route was assessed in patients with advanced cancer refractory to chemotherapy. Patients with progressive disease received escalating doses twice weekly. Treatment of the accrued 26 patients (10 colorectal cancer (CRC), 6 soft tissue sarcoma (STS), 5 melanoma (MM), 5 others) was well tolerated without substance-related grade 3 or 4 toxicities. Grade 1/2 toxicities were predominantly injection site reactions. Aviscumine lacked dose-limiting toxicity (DLT) up to a maximal dose of 10 ng/kg. An increase of interleukin-1 beta and interferon-gamma from baseline was seen in the patient's plasma between the 1st and 11th injection. Highest release of both cytokines was in the dose range of 4-5.9 ng/kg. Interferon-gamma was not detected after doses higher than 6 ng/kg. Eight patients (5 CRC, 1 MM, 1 STS, 1 RCC) had disease stabilisation for 79-250 days (median122 days) associated with an increase of interleukin (IL)-1 beta and interferon (IFN)-gamma. Aviscumine was well tolerated and appeared to possess clinical activity at a biologically active dose between 4 and 6 ng/kg.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Antineoplastic Agents, Phytogenic/administration & dosage , Neoplasms/drug therapy , Plant Preparations/administration & dosage , Ribosome Inactivating Proteins, Type 2/administration & dosage , Toxins, Biological/administration & dosage , Adjuvants, Immunologic/adverse effects , Adjuvants, Immunologic/pharmacokinetics , Administration, Cutaneous , Adult , Aged , Antibodies, Neoplasm/metabolism , Antineoplastic Agents, Phytogenic/adverse effects , Antineoplastic Agents, Phytogenic/pharmacokinetics , Dose-Response Relationship, Immunologic , Female , Humans , Infusions, Intravenous , Male , Maximum Tolerated Dose , Middle Aged , Plant Preparations/adverse effects , Plant Preparations/pharmacokinetics , Ribosome Inactivating Proteins, Type 2/adverse effects , Ribosome Inactivating Proteins, Type 2/pharmacokinetics , Toxins, Biological/adverse effects , Toxins, Biological/pharmacokinetics , Treatment OutcomeABSTRACT
This study investigates the effects of mistletoe lectin-I (ML-I) on melanoma growth and spread in vivo. The human melanoma cell line MV3 was xenografted into severe combined immunodeficient mice and vehicle solution or purified ML-I was administered at 30, 150 and 500 ng per kg body weight (20 mice per group) daily. After 19 days, mice were killed, primary tumours (PTs) and lungs were dissected out, and tumour weights, number of lung metastases (LMs), number of tumour-infiltrating dendritic cells (DCs), and apoptosis rates in the melanoma cells and in the DCs were assessed. A 35% reduction of PT weight (P=0.03) and a 55% decrease in number of LMs (P=0.016) were evident for low-dose ML-I (30 ng kg(-1)) treatment but not for higher doses. Mistletoe lectin-I increased apoptosis rates in the melanoma cells of PTs at all doses, while no induction of apoptosis was noted in the LMs. Low-dose ML-I significantly increased the number of DCs infiltrating the PTs (P<0.0001) and protected DCs against apoptosis, while higher doses induced apoptosis in the DCs (P<0.01). Our results demonstrate that low-dose ML-I reduced melanoma growth and number of metastases in vivo, primarily due to immunomodulatory effects.