Design, synthesis, antibacterial and anti-cell proliferation activities of 1,2,4triazino3,4-h 1,8naphthyridine-8-one-7-carboxylic acid derivatives / 药学学报

To discover novel fluoroquinolone lead compounds as possible anti-infective or/and antitumor chemotherapies, combination principle of pharmacophore-based drug design, a series of novel tricyclic fluoroquinolone title compounds, [1,2,4]triazino[3,4-h][1,8]naphthyridine-8-one-7-carboxylic acid derivatives ( 5a-5p), were designed and synthesized with a fused [1,2,4]-triazine ring unit. Their structures were characterized by spectral data and elemental analysis and the in vitro antibacterial and anti-cell proliferation activities were also evaluated. The results showed that the titled compounds exhibited more significant inhibitory activities against drug-resistant bacteria (Methicillin-resistant Staphylococcus aureus and multi drug-resistant Escherichia coli strains) and three tested cancer cell lines (human hepatoma SMMC-7721, murine leukemia L1210 and human murine leukemia HL60 cells). Interestingly, SAR showed that compounds with electron-donating groups attached to benzene ring had stronger antibacterial activity than antitumor activity, but electron-withdrawing compounds displayed more potential antitumor activity than antibacterial activity, especially antitumor activity of nitro compounds was comparable to comparison doxorubicin. Thus, novel triazine-fused tricyclic fluoroquinolones as potent anti-infective or/and antitumor lead compounds are valuable to pay attention and for further development.

Part IV. Synthesis and antitumor evaluation of s-triazolothiadiazines and pyrazolo s-triazoles derived from ciproxacin / 药学学报

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 < 25.0 micromo x L(-1)) than that of parent ciprofloxacin (IC50 > 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.

Part IV: Design, synthesis and antitumor activity of fluoroquinolone C-3 heterocycles: bis-oxadiazole methylsulfide derivatives derived from ciprofloxacin / 药学学报

To explore an efficient strategy for further development of anticancer fluoroquinolone candidates derived from ciprofloxacin, a heterocyclic ring as the bioisosteric replacement of C3 carboxyl group led to a key intermediate, oxadiazole thiol (5), which was further modified to the bis-oxadiazole methylsulfides (7a-7h) and the corresponding dimethylpiperazinium iodides (8a-8h), respectively. Structures were characterized by elemental analysis and spectra data, and their anticancer activities in vitro against CHO, HL60 and L1210 cancer cells were also evaluated by MTT assay. The preliminary results show that piperazinium compounds (8) possess more potent activity than that of corresponding free bases (7).

Part II: Design, synthesis and antitumor action of C3/C3 bisfluoroquinolones linked-cross 2, 5-1, 3, 4oxadiazole / 药学学报

To develop a new small molecular probe for discovering an antitumor lead compound from the replacement of carboxylic group of two molecular antibacterial fluoroquinolones with a heterocyclic ring, a series of the C3/C3 bis-fluoroquinolones tethered with an 1, 3, 4-oxadiazole ring were synthesized as their respective HCl salts, and their structures were characterized by elemental analysis and spectral data. The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated via the respective IC50 values by methylthiazole trazolium (MTT) assay.

Synthesis and antitumor activity of C3 heterocyclic-substituted fluoroquinolone derivatives (I): ciprofloxacin aminothiodiazole Schiff-bases / 药学学报

To discover a novel antitumor lead compound derived from fluoroquinolone, C3 carboxyl group of ciprofloxacin (1) was replaced with heterocyclic ring to form cyclopropyl fluoroquinolone aminothiadiazole scaffold (2), then reacted with aromatic aldehydes to give the Schiff bases compounds (3a-3j). The structures of new compounds were characterized by element analysis and spectral data, and their in vitro antitumor activity against SMMC-7721, HL60 and L1210 cell lines was evaluated by MTT assay via the respective IC50 values. The bioactive assay showed that eleven thiadiazole-substituted ciprofloxacin derivatives displayed potential cytotoxicity against the tested cancer cell lines, where the IC50 values of compounds 3d and 3f reached micromolar concentration. Therefore, the C3 carboxyl group of fluoroquinolone is not necessary to antitumor activity. Functionally modified heterocycle-substituted fluoroquinolone as potent antitumor lead compound is valuable for further study.

Synthesis and antitumor activity of anthracene-9-carbaldehyde amino-s-triazole Schiff-bases with side-chain of S-acetic acid / 药学学报

To find out a novel lead compound from heterocyclic amine Schiff bases for developing new antitumor agents, each of (4-amino-5-substituted-s-triazol-3-ylthio) -acetic acids 2a-j was condensed with anthracene-9-carbaldehyde to obtain Schiff-bases of [4-(anthracen-9-yl methylene) amino] -5-substituted-s-triazol-3-ylsulfanyl] -acetic acids 3a-j. The structures of new compounds synthesized were characterized by elemental analysis and spectral data, and in vitro antitumor activity was also evaluated against CHO, HL60 and L1210 cell lines by MTT assay.

Regulatory effect of resveratrol on JAK1/STAT3 signal transduction pathway in leukemia / 中国实验血液学杂志

The aim of this study was to explore the molecule mechanism of resveratrol antileukaemia. The mouse lymphocytic leukemia L1210 cells were cultured and the expressions of pJAK1 and pSTAT3 protein in L1210 cells were detected by immunohistochemistry and immunoprecipitation in vitro. The mouse model with L1210 leukemia ascites carcinoma was established and activities of singal transduction pathway molecules pJAK1 and pSTAT3 were measured by Western blot and immunohistochemistry assay in vitro. The results indicated that resveratrol could significantly inhibit the JAK1/STAT3 signal transduction pathway, down-regulate expressions of pJAK1 and pSTAT3 and reduce the phosphorylation of JAK1 and STAT3 in a dose-and time-dependent manner. It is concluded that the resveratrol can regulate signal transduction pathway and reduce the activation of JAK1/STAT3 tyrosine phosphorylation significantly, and therefore resveratrol shows chemotherapeutic potential to leukaemia.

Indoleamine 2, 3-dioxygenase expression in cells of human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia and therapeutic effect of its inhibitor 1-methyl tryptophan / 中国实验血液学杂志

The objective of this study was to investigate the expression and function of indoleamine 2, 3-dioxygenase (IDO) in leukemia. The IDO expressions in human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia (ALL) were detected by immunofluorescence staining. Constructed leukemia mouse model was used to observe whether the IDO inhibitor, 1-methyl tryptophan (1-MT), has any effect in treating leukemia. The experimental group were fed with 1-MT solution every day while the mice in control group had no further treatment. The results showed that the average ratios of IDO expression were 29.4 +/- 11.2% in M(5) patients and 24.7 +/- 7.96% in ALL patients respectively. After statistical test, IDO expression level in leukemia cells was significantly higher than that of normal mononuclear cells. The tumor decreased gradually in mice treated with 1-MT. At the terminal point of the experiment (88 days after vaccination), the average survival time in the experimental group was 42.3 days while the mice in control group only lived 15.1 days in average, which difference was statistically significant (P < 0.05). Some of the leukemia mice in the experimental group long-term survived without tumor (more than three months after vaccination). It is concluded that human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia (ALL) express IDO, and both can be treated by 1-MT in mice.

Anti-tumor effect of two adjuvants of CpG ODN on leukemic tumor in mouse models / 中国实验血液学杂志

To investigate the anti-tumor and side effect of CpG ODN 1826 and CpG ODN2006 as an adjuvants on leukemic tumor in mouse-models, an acute lymphocytic leukemic tumor in mouse model was established, then inoculated inactivated L1210 cells alone or with different vaccine adjuvants were injected subcutaneously into each DBA/2 model mouse at different times. The activities of mice, the tumor formation rate and the growth status of leukemic tumor were observed. The tumor was examined by pathologic section. The results showed that the vaccine of inactivated L1210 cells and CpG ODN 1826 could decrease the leukemic tumor formation rate, slow down the growth of leukemic tumor mass in mice and obviously cause necrosis of tumor cells, but it could not prolong the life spans of the tumor-burden mice; while CpG ODN2006 could not only decrease the tumor formation rate, slow down the growth of tumor mass in mice and result in obvious necrosis of tumor cells, but also could eliminate the existing tumor mass in mice, and prolong the life spans of the tumor-burden mice. It is concluded that using CpG ODN2006 as an adjuvant enhances the anti-tumor effect against the leukemic tumor, prolong the life span of tumor-burden mice without obvious side effect.

Synthesis of methotrexate-poly (ethylene glycol) conjugate and their anti-tumor activity in vitro / 药学学报

To improve the physical property and bioactivity of methotrexate, this paper investigated the new formation of conjugate methotrexate-poly (ethylene glycol) and in vitro anti-tumor activity of the synthesized conjugate. The conjugate of methotrexate-poly (ethylene glycol), which was verified by the spectroscopy analysis of UV, IR and 13C NMR, was synthesized by chemical catalysis and micro-wave irritation. The determination for the conjugate of solubility in water and distribution coefficient in octanol-water system of the conjugate was done to examine its deliquescence property. The solubility in water and the distribution coefficient of the conjugate was greatly improved, which was increased by 128 folds and 5 folds, respectively. The in vitro anti-tumor activity of the conjugate was tested by mouse L(1210) leukaemia cells, and the synthesized conjugate showed the same anti-tumor activity as the original methotrexate. Compared to the reported literature, the modification of methotrexate by poly (ethylene glycol) is more rapid and convenient.

Apoptosis-inducing effect of alternol on mouse lymphocyte leukemia cells and its mechanism / 药学学报

Alternol is purified from fermentation productions of microorganisms named as Alternaria alternata var. monosporus. The research is to investigate the apoptosis-inducing effect of alternol on mouse lymphocyte leukemia (L1210) cells and the possible mechanisms. MTT method was used to evaluate the viability of L1210 cells. Apoptosis of L1210 cells was detected by morphological assessment, DNA electrophoresis assay and flow cytometry. Western blotting analysis was carried out to determine the apoptosis-related proteins. Proliferation inhibition of L1210 cells by alternol was found remarkably in a dose-dependent manner. When treated with alternol, apoptotic morphological features of L1210 cells were observed by fluorescent microscopy (AO/EB) and the apoptosis rate was also elevated in a time-dependent manner. After treatments with various concentrations of alternol for 48 h, DNA laddering appeared. The increase of reactive oxygen species (ROS) production was found after cells were exposed to alternol for 6 h, while the decrease of mitochondrial transmembrane potential (delta psi m) was not found until cells were exposed to alternol for 24 h. Furthermore, the level of Bcel-2 and Bcl-2/Bax was down-regulated, while the level of caspase-3 and caspase-9 but not caspase-8 was up-regulated when alternol was added for 72 h. In summary, the results suggested that alternol could inhibit the proliferation of L1210 cells and induce apoptosis of L1210 cells, which was mediated by mitochondria-dependent pathway.

The effect of extremely low frequency magnetic fields on cytochrome oxidase subunit 1 mRNA transcription / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To clone and identify MF-1 gene which responded to extremely low frequency magnetic fields(ELF MF) in Daudi cells, and explore the response universality of MF-1 gene in several MF-sensitive cell lines, so as to provide experimental basis for revealing the mechanism of biological effects induced by magnetic field.</p><p><b>METHODS</b>The DNA fragment of MF-1 was cloned and sequenced; the mRNA level of MF-1 gene were analysed in MF-sensitive cell lines(HL-60, L1210 and CHL) by Northern blot after these cells being treated with 0.1 mT and 0.8 mT MF for 20 minutes and 24 hours, respectively.</p><p><b>RESULTS</b>The MF-1 cDNA sequence had 100% homology with cytochrome oxidase subunit 1 gene(CO1) by searching Gene Bank database; the transcription of CO1 in HL-60, L1210 and CHL cell lines which exposed to 0.1 mT and 0.8 mT MF for 20 minutes were significantly lower(0.38 +/- 0.12 and 0.37 +/- 0.04) than that of control(0.58 +/- 0.12) and so did for 24 hours exposure(0.46 +/- 0.09 and 0.45 +/- 0.09 vs 0.65 +/- 0.06) (P < 0.05).</p><p><b>CONCLUSION</b>CO1 is a MF-responsive gene. Cytochrome oxidase activity may be affected through low level of CO1 transcription by magnetic fields, thus induce bioeffects in organisms.</p>

Collateral methotrexate resistance in cisplatin-selected murine leukemia cells

Resistance to anticancer drugs is a major cause of failure of many therapeutic protocols. A variety of mechanisms have been proposed to explain this phenomenon. The exact mechanism depends upon the drug of interest as well as the tumor type treated. While studying a cell line selected for its resistance to cisplatin we noted that the cells expressed a >25,000-fold collateral resistance to methotrexate. Given the magnitude of this resistance we elected to investigate this intriguing collateral resistance. From a series of investigations we have identified an alteration in a membrane protein of the resistant cell as compared to the sensitive cells that could be the primary mechanism of resistance. Our studies reviewed here indicate decreased tyrosine phosphorylation of a protein (molecular mass = 66) in the resistant cells, which results in little or no transfer of methotrexate from the medium into the cell. Since this is a relatively novel function for tyrosine phosphorylation, this information may provide insight into possible pharmacological approaches to modify therapeutic regimens by analyzing the status of this protein in tumor samples for a better survival of the cancer patients

Effect of Multidrug Resistance Gene-1 (mdr1) Overexpression on In-Vitro Uptake of 99mTc-sestaMIBI in Murine L1210 Leukemia Cells / 대한핵의학회잡지

PURPOSE: To determine whether Tc-99mMIBI is recognized by the multidrug resistant P-glycoprotein (Pgp), we have measured quantitatively Tc-99mMIBI uptake in cancer cells. The effects of various Pgp reversing agents on cellular Tc-99m-MIBI uptake were also investigated in the presence of multidrug resistance gene-1 (mdr1 gene) overexpression. MATERIALS AND METHODS: We measured percentage uptake of Tc-99m-MIBI at different incubation temperatures both in mdr1 positive and negative cells. The effects of verapamil, cyclosporin, and dipyridamole on cellular uptake of Tc-99m-MIBI were also evaluated with or without overexpression of mdr1 gene in cultured murine leukemia L1210 cells. RESULTS: The mdr1 gene expressing cell lines were effectively induced in in vitro with continuous application of low-dose adriamycin or vincristine. Cellular uptake of Tc-99m-MIBI was higher in mdr1 negative L1210 cells than those of mdr1 positive cells, and higher when incubated in 37 degree C than 4 degree C. In the presence of verapamil, cyclosporin or dipyridamole, Tc-99m-MIBI uptake was increased upto 604% in mdr1 positive cells. CONCLUSION: Cellular uptake of Tc-99m-MIBI is lower in leukemia cells over-expressing mdr1 gene, and MDR-reversing agents increase cellular uptake. These results suggest that Tc-99m-MIBI can be used for characterizing Pgp expression and developing MDR-reversing agents In vitro.

Cloning of adriamycin-resistant related (arr) gene in an adriamycin-resistant L1210 variant

A partial fragment of novel sequence (arr, adriamycin-resistant related) was previously identified using the differential display (DD)-PCR technique with adriamycin-resistant L1210 variant (L1210AdR), which shows a typical multidrug resistant (MDR) phenotypes. The present research shows the isolation of full length arr cDNA sequence. To clone the full length cDNA of arr gene, DD-PCR fragments were subjected to 5'- and 3'-Rapid Amplification of cDNA End (RACE) method. The cloned arr cDNA consisted of 770 bases and contained an open reading frame of 153 bases, encoding a protein of 51 amino acid with the molecular mass of 4 kDa by in vitro translation reactions. Northern blot analysis showed that a 770 bases transcript arr gene was overexpressed in adriamycin-resistant L1210 variant, but not in the parent suggesting that the arr gene may be involved in the adriamycin-resistant phenotypes.

Analogues and prodrugs, permanent useful alternatives to improve the effectiveness of natural anticancer compounds: Enhanced activity of slightly soluble salts of olivacine in L1210 leukemia

Several derivatives of well-known natural anticancer drugs have been synthesized and will continue to be, to overcome their delivering problems, or effectiveness. Characteristics such as poor solubility, metabolic inactivation, resistance development, rapid clearance or short half-life, adverse side effects like nausea, alopecia, anorexia are common undesirable limitations to their clinical use. Drug toxicity to less sensitive tumors as those in liver or brain is another aimed goal. Recent efforts to reverse undesirable physicochemical properties of well-established natural anticancer compounds are focused. Improved antitumor action of olivacine in L(1210) leukemia with slightly soluble halides with increased life span as well as "cured" animals was observed.

Actividad antitumoral del CCNU obtenido en Cuba / Antineoplasic activity of CCNU obtained in Cuba

El (1-[2-cloroetil] 1-nitroso, 3 ciclohexil urea) (CCNU) es una droga antitumoral que forma parte actualmente de múltiples esquemas de poliquimioterapia en el tratamienmto de diferentes localizaciones y su obtención en el país aumentará la disponibilidad de este producto. El presente trabajo evalúa la actividad, en comparación con el producto comercial usado actualmente en la clínica oncológica y por los resultados obtenidos en los tumores experimentales utilizados, ambos productos manifiestan semejante actividad antitumoral

Sinergismo del Cis-Diaminodicloro Platino (II) (CIS-DDP) y el M-Tiocarbamal en el tratamiento de la leucemia L-1210 / Synergism of Cis-Platinum Diamminodichloride (II) (CIS-DDP) and M-Thiocarbamol in the treatment of leukemia L-1210

El M-tiocarbamal (morfolinditiocarbamato de sodio) es un producto sintético con el cual en estos momentos se lleva a cabo el ensayo clínico fase I. Experimentalmente se combinó con el Cis-DDP en el tratamiento de la leucemia L-1210. Los resultados obtenidos evidencian la potenciación de la acción antitumoral del Cis-DDP cuando el M-tiocarbamal se administra simultáneo o después de la administración del citostático

Effects of cimetidine combination with cyclophosphamide in transplanted murine tumors.

Studies were carried out on the combination of Cimetidine (CMTD) with Cytoxan (CTX) in three murine tumors. While the combination significantly potentiated the anticancer effect of CTX in L1210 leukemia, the results with P388 leukemia were not significantly different. The results with Lewis Lung Carcinoma showed a consistent reduction in the number of metastases. However, there was no consistent concomitant prolongation in survival. The host strain, biology of the tumour and the drug used in combination with CMTD might be some of the factors responsible for the varied response.

Actividad antitumoral y principales efectos tóxicos del 1,3 biscloroetil nitrosourea (BCNU) sintetizado en Cuba / Antitumoregenic activity and main toxic effects of 1,3 bischloroethylnitrosourea (BCNU) synthetized in Cuba

Se estudió la actividad antitumoral del 1,3 biscloroetil nitrosourea (BCNU) sintetizado en Cuba en dos tumores transplantables de ratón, las leucemias L-1210 y P-388, así como la toxicidad de diferentes esquemas de administración del producto en ratas albinas. El producto aumentó notablemente la supervivencia de los animales tratados con respecto al control en los tumores experimentales usados y manifestó efectos tóxicos notables en el sistema hematopoyético y el hígado