两类氮芥衍生物的合成及其生理活性研究

设计合成了3个苯胺氮芥和6个萘酰亚胺-氮芥衍生物,产物结构均经1H NMR,13C NMR,ESI-MS和元素分析确认.使用MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]法测试了9个化合物对HL-60,K562和SMMC-7721三种...     

Design and Synthesis of Glycosylated Aromatic Nitrogen Mustard Derivatives

Antibody-directed enzyme prodrug therapy(ADEPT) is a new strategy for the treatment of cancer that has arisen in recent twenty years, the main merits of which are that it can improve the selectivity of anticancer drugs and reduce the side effects in remote tissue. In the present study, two prodrugs-glycosylated aromatic nitrogen mustard derivatives were synthesized. Glucose and lactose were converted into glycosyl donors-trichloroacetimidate; the obtained glycosyl donors were glycosylated with p-nitrophenol(glycosyl donors) to form β-glucosyl p-nitrobenzene and β-lactosyl p-nitrobenzene that were protected by acetyl in a stereoselective manner; the two products were reduced by zinc dust and then treated with ethylene oxide, afforded two glycosylated nitrogen mustard derivatives that were protected by acetyl; the last step was to deacetylate and then afforded the two target compounds that could be used as prodrugs of ADEPT for further Anti-tumor research.     

Synthesis,cytotoxicity, topoisomerase I inhibition and molecular docking of novel phosphoramide mustard sophoridinic acid analogues

A series of novel phosphoramide mustard sophoridinic acid analogues, consisting of nitrogen mustard group and sophoridinic acid scaffold, have been designed, synthesized and evaluated for their topoisomerase inhibitory activity as well as cytotoxicity against six tumor cell lines (SMMC‐7721, LoVo, MCF‐7, K562, S180 and H22) and a normal cell line (L929). Among the compounds tested, five were found to be potent inhibitors and exhibited potent cytotoxicity against S180 and H22 cell lines with IC₅₀ values of 1–4 μM. Further mechanistic studies showed that this class of compounds acted as novel topoisomerase I (Topo I) catalytic inhibitors by preventing the binding of Topo I to DNA and inhibiting the cleavage of DNA, and molecular docking studies revealed that the binding energy for these compounds was comparable to that for classic Topo I inhibitors CPT and HCPT, indicating that the compounds have an interaction with DNA and Topo I.     

长链醇和胺修饰的芳香氮芥衍生物的合成及生理活性

为了研究不同取代基对苯甲酸氮芥抗肿瘤活性的影响,以对氨基苯甲酸为起始原料,设计合成了6个新化合物,目标产物结构经元素分析、1H NMR和MS等确证.采用MTT法对白血病细胞HL-60、白血病细胞K562和肝癌细胞SMMC-7721进行了体外的抗肿瘤活性筛选.活性测试结果表明这些长链醇和胺修饰苯甲酸氮芥后,目标产物的抗肿瘤活性未得到提高,均弱于参照药物美法伦.     

The cellular labeling and pH-sensitive responsive-drug release of celastrol in cancer cells based on Cys-CdTe QDs

As one of the active compounds derived from Traditional Chinese Medicine,Celastrol(CSL)had cytotoxicity for human leukemia cancer cells K562 and its multidrug-resistant cell line K562/A02.Here,we introduced cysteamine-modified CdTe QDs as the labeling and drug carrier into CSL research and found that the self-assembly and conjugation of anticancer molecular CSL with the Cys-CdTe QDs could significantly increase the drug’s cytotoxicity for K562 cells.More important,these CSL-Cys-CdTe nanocomposites could overcome the multidrug resistance of K562/A02 cells and efficiently inhibit the cancer cell proliferation by realizing the pH-sensitive responsive release of CSL to cancer cells.The enhanced cytotoxicity was caused by the increase of the G2/M phase arrest for K562/A02 cells as well as for K562 cells.Cys-CdTe QDs can readily bind on the cell plasma membranes and be internalized into cancer cells to trace and detect human leukemia cancer cells in real time.In addition,these Cys-CdTe QDs can facilitate the inhibition of the multidrug resistance of K562/A02 cells and readily induce apoptosis.As a good photosensitizer for the therapy,labeling,and tracing of cancer cells,the combination of CSL with Cys-CdTe QDs can optimize the use of and a new potential therapy method for CSL and yield new tools to explore the mechanisms of active compounds from Traditional Chinese Medicine.     

硒化壳聚糖抗K562细胞的实验研究

用MTT法和AO/EB荧光染色法观察了硒化壳聚糖对K 562肿瘤细胞株生长的影响。结果发现,硒化壳聚糖可有效地抑制K 562细胞生长,并呈量效、时效关系。经硒化壳聚糖作用后的细胞可明显出现核固缩、碎裂等凋亡形态改变。硒化壳聚糖可诱导K 562细胞凋亡,抑制其生长。     

Design,synthesis, and biological evaluation of sorafenib derivatives containing indole (ketone) semicarbazide analogs as antitumor agents

A series of new sorafenib derivatives was designed and synthesized. The antiproliferative activity of the synthesized compounds against human lung cancer cell (A549), human pancreatic cancer cell (PC-3), human leukemia cell (K562), and human hepatoma cell (SMMC-7721) was evaluated by MTT assay. The results revealed that several compounds displayed more significant antitumor activities than commercial anticancer agent sorafenib against SMMC-7721. In addition, compounds 7a , 7g , 7l , 7m , and 7p represented obvious growth inhibition with IC₅₀ values of 1-9 μM against four cancer cell lines, demonstrating more predominant activities against cancer cells as compared to sorafenib. Furthermore, some structure-activity relationships have also been established. Compounds containing indole and benzene ring substituted by halogen showed better activity than sorafenib. Wound healing assay suggested that cells would be targeted on their migratory capacity by 7g , potentially affecting the migration activity of these tumors. The effects of A549 and PC-3 cell apoptosis induced by compound 7g were significantly increased compared with sorafenib. Importantly, the result of western blot assay showed that 7g inhibited cell growth by suppressing the activity of EGFR, especially the expression of p-EGFR (Tyr1068).     

茄呢基氮芥衍生物的合成及生理活性

茄呢基氮芥衍生物的合成及生理活性;氮芥;茄呢醇;合成;生理活性     

单独和合用硒化壳聚糖与阿霉素对K562细胞作用的研究

应用MTT法和AO／EB荧光染色法观察单独和合用硒化壳聚糖与阿霉素对K562细胞株的影响,结果发现硒化壳聚糖和阿霉素均可有效地抑制K562细胞生长。且两药联合使用效果更佳。硒化壳聚糖可诱导细胞凋亡。两药合用诱导细胞凋亡效果更好。     

8-取代黄酮的合成及生理活性研究

设计合成了8-黄酮甲基氮芥和6种8-黄酮甲基多聚异戊二烯胺类化合物, 产物结构经¹H NMR, MS及元素分析确证. 合成的7个化合物与氮芥类抗肿瘤药物美法仑一起对白血病细胞(K562)、中国仓鼠卵巢细胞(CHO)、黑色素瘤细胞(B16)等3种肿瘤细胞进行了初步的体外生理活性测试. 结果表明, 对3种受测细胞, 8-黄酮甲基氮芥的体外活性均与美法仑接近; N,N-二(8-黄酮甲基)香叶基胺则表现出比美法仑更高的体外抗肿瘤活性.     

Synthesis and Characterization of Thiophene‐derived Amido Bis‐nitrogen Mustard and Its Antimicrobial and Anticancer Activities

The thiophene‐derived amido bis‐nitrogen mustard N²,N²,N⁵,N⁵‐tetrakis(2‐chloroethyl)‐3,4‐dimethylthiophene‐2,5‐dicarboxamide was designed and synthesized via five‐step reactions from commercially available 2‐chloroacetonitrile. This target compound was confirmed by ¹H NMR, ¹³C NMR, MS, IR spectra and elemental analyses, and its structure was further characterized by X‐ray single‐crystal analysis. The biological activities for the title compound and some intermediates were evaluated in vitro for their antibacterial, antifungal and cytotoxic activities. The preliminary results showed that the title compound could inhibit efficiently the growth of the tested microorganisms including drug‐resistant bacteria MRSA to some extent. Moreover, the target compound was found to be effective against prostatic carcinoma cell line (PC‐3), breast carcinoma cell line (MCF‐7), colon carcinoma (LoVo) and lung cancer (A549). Especially, it gave selective antitumor efficacy against prostatic carcinoma cell line (PC‐3) at a low dose.     

Anti-lipid peroxidation and induction of apoptosis in the erythroleukaemic cell line K562 by extracts from (Tunisian) Rhamnus alaternus L. (Rhamnaceae)

Total oligomer flavonoids (TOF) enriched and ethyl acetate (EA) extracts from Rhamnus alaternus induce apoptotic death in human chronic myelogenous leukaemia K562 cell line, as demonstrated by gel electrophoresis, which demonstrates the characteristic ladder patterns of DNA fragmentation and the proteolytic cleavage of poly(ADP ribose) polymerase (PARP). The effect of R. alaternus extract in reducing oxidative stress was evaluated by anti-lipid peroxidation which was monitored by measuring malondialdehyde level in K562 cultured cells. The TOF and EA extracts were found to be effective to protect against lipid peroxidation. Their IC?? values were 196 and 273?μg?mL?1, respectively. These findings suggest that R. alaternus extracts exhibit potential antioxidant and proapoptotic properties.     

Aromatic Nitrogen Mustard‐Based Prodrugs: Activity,Selectivity, and the Mechanism of DNA Cross‐Linking

Three novel H₂O₂‐activated aromatic nitrogen mustard prodrugs ( 6 – 8 ) are reported. These compounds contain a DNA alkylating agent connected to a H₂O₂‐responsive trigger by different electron‐withdrawing linkers so that they are inactive towards DNA but can be triggered by H₂O₂ to release active species. The activity and selectivity of these compounds towards DNA were investigated by measuring DNA interstrand cross‐link (ICL) formation in the presence or absence of H₂O₂. An electron‐withdrawing linker unit, such as a quaternary ammonia salt ( 6 ), a carboxyamide ( 7 ), and a carbonate group ( 8 ), is sufficient to deactivate the aromatic nitrogen mustard resulting in less than 1.5 % cross‐linking formation. However, H₂O₂ can restore the activity of the effectors by converting a withdrawing group to a donating group, therefore increasing the cross‐linking efficiency (>20 %). The stability and reaction sites of the ICL products were determined, which revealed that alkylation induced by 7 and 8 not only occurred at the purine sites but also at the pyrimidine site. For the first time, we isolated and characterized the monomer adducts formed between the canonical nucleosides and the aromatic nitrogen mustard ( 15 ) which supported that nitrogen mustards reacted with dG, dA, and dC. The activation mechanism was studied by NMR spectroscopic analysis. An in vitro cytotoxicity assay demonstrated that compound 7 with a carboxyamide linker dramatically inhibited the growth of various cancer cells with a GI₅₀ of less than 1 μM , whereas compound 6 with a charged linker did not show any obvious toxicity in all cell lines tested. These data indicated that a neutral carboxyamide linker is preferable for developing nitrogen mustard prodrugs. Our results showed that 7 is a potent anticancer prodrug that can serve as a model compound for further development. We believe these novel aromatic nitrogen mustards will inspire further and effective applications.     

Synthesis and characterization of glycosylated nitrogen mustard derivatives and their interaction with bovine serum albumin

To improve the specificity of nitrogen mustards towards tumor cells, glucose-nitrogen mustard, fructose-nitrogen mustard, and lactose-nitrogen mustard were prepared as three novel glycosylated nitrogen mustard derivatives by esterification of bis(2-chloroethyl)carbamic chloride (BCC) with glucose, fructose, and lactose, respectively. BCC was synthesized from bis(2-chloroethyl)amine hydrochloride and triphosgene. The topic products were characterized by infrared (IR) and mass spectrometry (MS), and their interaction with bovine serum albumin was investigated by measuring fluorescence spectra in tris(hydroxymethyl)aminomethane hydrochloride (Tris–HCl) buffer solution at physiological conditions.     

Synthesis,characterization, and biological studies of lanthanide complexes with 2,6-pyridine dicarboxylic acid and α-picolinic acid

Four new solid ternary complexes of lanthanide with 2,6-pyridine dicarboxylic acid and α-picolinic acid [Ln(DPA)(L^α)(H₂O)] · 2H₂O (Ln = La³⁺, Ce³⁺, Eu³⁺, or Gd³⁺; DPA = 2,6-pyridine dicarboxylic acid; HL^α = α-picolinic acid) have been synthesized and characterized by elemental analysis, molar conductance, FT-IR, UV–Vis, and TG–DTA. The antibacterial activities indicate that all the complexes exhibit antibacterial ability against Escherichia coli and Staphylococcus aureus with broad antimicrobial spectra. The anticancer activity of the La complex against K562 tumor cell in vitro is measured using methyl thiazolyl tetrazolium (MTT) colorimetry and flow cytometry. The La complex can induce K562 tumor cell apoptosis, presenting the best apoptosis effect by acting on the S period after inducing K562 tumor cell for 72 h.     

Preferential cytotoxicity for multidrug-resistant K562 cells using the combination of a photosensitizer and a cyanine dye

The cyanine dye 1,1',3,3,3',3'-hexamethylindodicarbocyanine iodide (HIDC) protects K562 leukemia cells from photodynamic membrane damage caused by cis-di(4-sulfonatophenyl)diphenylporphine (TPPS2) and 420 nm light. This wavelength of light is chosen because it is absorbed by TPPS2, but not by HIDC. The photodynamic system studied may be useful as a model for antineoplastic therapy. A subline of K562 leukemia (K562/DOX), expressing the multidrug-resistance (MDR) phenotype, is found to accumulate smaller amounts of HIDC than the parent cell line and thus has less photoprotection. In the absence of added HIDC, the K562/DOX cell line is more resistant to photodynamic cytotoxicity than the K562 cell line. The resistance of the K562/DOX cell line is not due to a smaller accumulation of TPPS2 than the K562 cell line. However, when both cell lines are incubated with HIDC and TPPS2, and then exposed to light, the K562/DOX cell line becomes more sensitive to photodynamic cell damage than the K562 cell line. The combination of a photosensitizer with a cationic or lysomorphotropic photoprotector represents a novel strategy for the eradication of malignant cells expressing the MDR phenotype.     

茄呢基胺-氮芥衍生物的合成

首次设计并制备了二种以茄呢基胺和茄呢基哌嗪为载体的新型氮芥衍生物.即以二 (2 氯乙基 )胺盐酸盐为原料与邻苯二甲酸酐反应,合成出含有抗肿瘤活性基团的化合物 2 { [二 ( 2 氯乙基 )氨基 ]羰基 }苯甲酸,然后在二环己基碳二亚胺存在下,分别与茄呢基胺和茄呢基哌嗪进行酰化反应,得到两种含茄呢基胺类基团的氮芥衍生物,其结构经元素分析,IR,1HNMR和MS确证.     

Furocoumarins photolysis products induce differentiation of human erythroid cells

Psoralens, also known as furocoumarins, are a well-known class of photosensitizers largely used in the therapy of various skin disease. In this study we have evaluated the effects of crude pre-irradiated solutions of furocoumarins derivatives on (a) erythroid differentiation and apoptosis of human leukemic K562 cells and (b) hemoglobin synthesis in cultures of human erythroid progenitors derived from the peripheral blood. To prove the activity of a mixture of photoproducts generated by UVA irradiation of the three psoralen derivatives 5-methoxypsoralen (5-MOP) 8-methoxypsoralen (8-MOP), and angelicin (ANG), we employed the human leukemic K562 cell line and the two-phase liquid culture procedure for growing erythroid progenitors. The results obtained demonstrate that pre-irradiated solutions of psoralen derivatives significantly induce erythroid differentiation of K562 cells irrespective of the type of derivative used, suggesting that the active photoproduct(s) share a common structure. Interestingly, solutions of psoralens irradiated in anaerobic conditions do not exhibits erythroid inducing ability, indicating that the effect is mostly due to photooxidized psoralen products. In erythroid precursor cells, psoralens photolysis products stimulates at low concentrations an increase of hemoglobin A and hemoglobin F. Altogether, these data suggest that photoproducts of psoralen warrant further evaluation as potential therapeutic drugs in beta-thalassaemia and sickle cell anaemia.     

Blocking ACAT‐1 Activity for Tumor Therapy with Fluorescent Hyperstar Polymer‐Encapsulated Avasimible

Targeting the distinct cholesterol metabolism of tumor cells is proposed as a novel way to treat tumors. Blocking acyl‐CoA cholesterol acyltransferase‐1 (ACAT‐1) by the inhibitor avasimible (Ava), which elevates intracellular free cholesterol levels, is shown to effectively induce apoptosis. However, Ava faces disadvantages of poor water solubility, a short half‐life, and no capability for fluorescence detection, which have greatly limited its application. Herein, a fluorescent hyperstar polymer (FHSP) is developed to encapsulate Ava to improve its ability to inhibit HeLa cells and K562 cells. The results of this study show that the obtained Ava–FHSP micelles possess a high drug loading capacity of 22.7% and bright green fluorescence. Ava and Ava–FHSP are cytotoxic to both HeLa and K562 cells and cause reductions in cell size, nuclear lysis, and chromatin condensation and hindered proliferation of both cell types by causing S phase cell cycle arrest. Further mechanistic analysis indicates that Ava–FHSP reduces the protein and messenger RNA expression of ACAT‐1 and significantly increases intracellular free cholesterol levels, which can increase endoplasmic reticulum stress and finally cause cell apoptosis. All these results suggest that this fluorescent hyperstar polymer represents a potential therapeutic tumor strategy by changing the cholesterol metabolism of tumor cells.     

Synthesis, Crystal Structure and Inhibition of the K562 Cell Proliferation of N＇-Tertbutylaminocarbonyl-N-2-chlorophenoxyacetylthiourea

The title compound N'-tert-butylaminocarbonyl-N-2-chlorophenoxyacetylthiou- rea has been synthesized for the first time. Complete assignments were achieved by IR, 1H NHR and single-crystal X-ray diffraction analyses. The inhibitory rate of the cellular growth of K562 cells (chronic myeloid 1eukemic cells) was measured using MTT [3-(4,5-dimethylthiazo-2-y1)-2,5-di- phenyltetra-zolium bromide] assay. The cell apoptosis was assessed by agarose gel electrophoresis to find that the title compound has antiproliferation and apoptosis inducing effects on K562 cells. In order to investigate the relationship between structure and activity of the target compound, we report its crystal structure and biological behavior in the present paper. Crystallographic data: C14H18- ClN3O3S, Mr = 343.82, orthorhombic, space group Pnma, a = 19.786(6), b = 6.789(2), c = 12.938(4) , V = 1738.0(9) 3, Z = 4, Dc = 1.314 g/cm3, F(000) = 720, μ(MoKα) = 0.354 mm-1, R = 0.0378 and wR = 0.0941. The molecule is a planar structure.