CN128: A New Orally Active Hydroxypyridinone Iron Chelator.

Deferoxamine, deferiprone, and deferasirox are used for the treatment of systemic iron overload, although they possess limitations due to lack of oral activity, lower efficacy, and side effects. These limitations led to a search for an orally active iron chelator with an improved therapeutic index. The lower efficacy of deferiprone is due to rapid glucuronidation, leading to the formation of a nonchelating metabolite. Here, we demonstrate that the influence of metabolism can be reduced by introducing a sacrificial site for glucuronidation. A log P-guided investigation of 20 hydroxpyridinones led to the identification of CN128. The Fe(III) affinity and metal selectivity of CN128 are similar to those of deferiprone, the log P value is more lipophilic, and its iron scavenging ability is superior. Overall, CN128 was demonstrated to be safe in a range of toxicity assessments and is now in clinical trials for the treatment of ß-thalassemia after regular blood transfusion.

[The effect of ferric ion on the differentiation of osteoclast and bone resorption].

OBJECTIVE: To explore the effects of ferric ion on the differentiation from both RAW264.7 and bone marrow macrophages to osteoclast in vitro and bone resorption in vivo. METHODS: In the presence of 50 ng/ml receptor activator of nuclear factor kappa-B ligand (RANKL), RAW264.7 was treated with ferric ammonium citrate (FAC). The formation of osteoclast was observed by staining of tartrate-resistant acid phosphatase (TRAP) and the TRAP positive cell counted. The expression levels of TRAP, cathepsin-K, nuclear factor of activated T cells c1 (NFATc1) and (receptor activator of NF-κB) RANK were measured by reverse transcription-polymerase chain reaction (RT-PCR).The control and iron overload groups were established by the intraperitoneal injection of normal saline and FAC. In vivo imaging system was employed to determine the bone density of femoral midportion and the fourth lumbar vertebra. After that, the bone marrow cells of femurs were used for osteoclast culture. The serum levels of ferritin, TRAP-5b, RANKL, osteoprotegerin (OPG) and C-terminal cross-linking telopeptide (CTX) were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Ferric ion could stimulate the formation of TRAP positive cells in a dose-dependent manner. The expression levels of TRAP, cathepsin-K and NFATc1 in the FAC treated group were significantly higher those of the control group (P < 0.05) while the expression of RANK showed no statistical difference among these groups (P = 0.967). The bone marrow density of femoral midportion and the fourth lumbar vertebra of the iron-overload group decreased significantly versus the control group. The concentrations of ferritin, TRAP-5b, RANKL and CTX of the iron overload group were markedly higher than those of the control group (P < 0.05). Moreover, the concentration of ferritin showed a positive correlation with TRAP-5b and CTX respectively in the iron-overload group (r = 0.65, r = 0.76, P < 0.05). But no significant differences existed in the concentration of OPG for two groups (P > 0.05). CONCLUSION: Ferric ion may enhance the differentiation of osteoclast in vitro as well as bone resorption in vivo.

Cigarette smoke-induced failure of apoptosis resulting in enhanced neoplastic transformation in human bronchial epithelial cells.

The lack of apoptotic pathways may lead to undesirable cell survival and proliferation, which are recognized hallmarks of cancer. It is well known that exposure to cigarette smoke induces DNA lesions in pulmonary cells. At present, it is not fully elucidated whether these lesions are repaired to restore normal functions or induce apoptosis. In order to examine the role of apoptosis in smoking-induced effects, immortalized human bronchial epithelial cells (BEAS-2B) were exposed to cigarette smoke and examined for parameters associated with apoptosis and neoplastic transformation. Our results indicated a significant reduction in apoptosis and enhanced neoplastic transformation and decreased mitochondrial membrane potential Δψm of mitochondria compared to control cells. Time-course experiments revealed increased aberrant methylation of CpG islands of RAS-associated domain family protein 1A (RASSF1A) and O (6)-methylguanine-DNA-methyltransferase (MGMT). The activities were downregulated and repair of DNA adducts was inhibited. Our observations suggested that although cigarette smoke-induced damage in BEAS-2B cells after chronic exposure is not necessarily lethal, as evidenced by cell viability, the protein expression levels of caspase-3 showed a decrease in the S20 passage (metaphase) but subsequently increased from S30 to S40 (anaphase). Survivin expression was significantly changed in S5 cells, and this rise was maintained until S40. Our data suggest that the potency of cigarettes as carcinogens may be due to their ability to induce aberrant gene expression and failure to trigger apoptosis leads to subsequent neoplastic transformation.

[Radiation protection and possible mechanisms for low intensity microwave].

OBJECTIVE: To investigate radiation protection and possible mechanisms of low intensity microwave on gamma-ray exposed mice. METHODS: 96 healthy Kunming mice were randomly divided into the following four groups: normal control, microwave (120 microW/cm(2), 900 MHz), gamma-ray irradiation (5 Gy), combined exposure of microwave and gamma-ray (120 microW/cm(2) + 5 Gy). The microwave group and combined group were exposed to 120 microW/cm(2) microwave firstly, 1 h/d, for 14 days. Then the ionization and combined group were exposed to 5 Gy (60)Co gamma-ray irradiation on the 15th day. Animals were sacrificed on the third, 6th, 9th and 12th day after irradiation. The sternum and spleen paraffin section were produced, and the histological changes were observed. Apoptosis rate of mice splenic cells in each group was examined by flow cytometry, and serum concentration of antioxidant and lipid peroxide was detected at the same time. RESULTS: Bone marrow was obviously injured either by radiation or microwave exposure, characterized by undergoing four-phase lesions, namely apoptosis-necrosis, void, regeneration and recovery phase. Compared with the gamma-ray group, the pathological changes in combined group were slighter and the recovery was quicker. The pathological injuries of spleen were similar to that of bone marrow. Injuries in the combined group were slighter than gamma-ray group. It showed that apoptosis rate of splenic cells in combined group was significantly lower on the 6th and 9th day after gamma-ray radiation (23.02% +/- 15.18%, 25.37% +/- 11.62% respectively) from FCM results. Assays of oxidative damages suggested that serum superoxide dismutase (SOD) level in combined group increased while lipid peroxide level decreased significantly (P < 0.05). CONCLUSION: Low intensity microwave may exert protection effects on injuries induced by ionizing radiation. The underlying mechanisms might be related with suppression on the hematopoietic cells apoptosis induced by gamma-ray radiation, inhibition of oxidative damages, and thus enhanced reconstruction of the hematopoietic system.

[Antagonistic effect of microwave on hematopoietic damage of mice induced by gamma-ray irradiation].

OBJECTIVE: To investigate antagonistic effect of microwave on hematopoietic damage of mice induced by gamma-ray irradiation. METHODS: Male healthy Kunning mice were treated with low dose microwave radiation before exposure to (60)Co gamma-ray irradiation of 8.0 Gy. The 30-day survival rate and average survival time of the mice after the treatment were examined. Peripheral blood parameters and the organ indexes of thymus and spleen were also observed in the irradiated mice. After exposure to 5.0 Gy gamma irradiation, indexes of hematopoietic foci formation of bone marrow cells (CFU-GM) and the proliferation activity of BMNCs were examined. The serum concentration of hemopoietic factors (GM-CSF and IL-3) were detected by ELISA kits. RESULTS: Pre-exposure with 120 microW/cm(2) 900 MHz microwave increased the 30-day survival rate (P < 0.05) and the number of white blood cells of gamma-ray treated mice. The increases of the organ indexes of thymus and spleen, proliferation activity of BMNCs and CFU-GM hematopoietic foci numbers, as well as the higher serum concentration of GM-CSF and IL-3 were observed in the microwave pre-exposure group. CONCLUSION: Low dose microwave radiation may exert potential antagonistic effects on hematopoietic injuries induced by ionizing radiation. The underlying mechanisms might be related with stimulation of hematopoietic growth factors expression, promotion of HSCs/HPCs proliferation, suppression on the reduction of HSCs/HPCs caused by (60)Co gamma-ray, and enhanced construction of the hematopoietic system.