Anton Kril

Synthesis, cytotoxicity and clastogenicity of novel α-aminophosphonic acids

Summary.α-Ethyl-N-(phosphonomethyl) glycine is synthesized and characterized by NMR and FAB spectroscopy. The cytotoxicity, clastogenic and antiproliferative effect of 3-ethyl-2-hydroxyl-2-oxo-1,4,2-oxazaphosphorinane, sodium salt of 3-ethyl-2-hydroxyl-2-oxo-1,4,2-oxazaphosphorinane, α-ethyl-α-N-(hydroxyethylamino) methylphosphonic acid, α-ethyl-N-(phosphonomethyl) glycine, α-ethyl-N-(phosphonomethyl) glycine isopropylammonium salt, glyphosate isopropylammonium salt are tested.

Synthesis, antiproliferative activity and genotoxicity of novel anthracene-containing aminophosphonates and a new anthracene-derived Schiff base.

A new Schiff base, 9-anthrylidene-furfurylamine and three novel anthracene-containing α-aminophosphonates, [N-methyl(dimethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine, [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine and [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]furfurylamine were synthesized. The compounds have been characterized by elemental analysis, TLC, IR, NMR and fluorescent spectra. The aminophosphonates and their synthetic precursors were tested for in vitro antitumor activity on a panel of seven human epithelial cancer cell lines. Safety testing was performed both in vitro (3T3 NRU test) and in vivo on ICR mice for genotoxicity and antiproliferative activity. 9-Anthrylidene-furfurylamine and [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]furfurylamine were most potent cytotoxic agents towards colon carcinoma cell line HT-29. The latter compound exhibited also antiproliferative activity to HBL-100, MDA-MB-231 and 647-V cells. The aminophosphonate [N-methyl(dimethoxyphosphonyl)-1-(9-anthryl)]-p-toluidine and its synthetic precursor 9-anthrylidene-p-toluidine were found to be cytotoxic to HBL-100 and HT-29 tumor cell lines, respectively. Moderate genotoxic and antiproliferative activity in vivo and low toxicity to Balb/c 3T3 (clone 31) mouse embryo cells were observed for all tested compounds. The subcellular distribution of two tested compounds in a tumor cell culture system was also studied.

Synthesis, characterization, antitumor activity and safety testing of novel polyphosphoesters bearing anthracene-derived aminophosphonate units

Novel polyphosphoesters containing anthracene-derived aminophosphonate units, poly(oxyethylene aminophosphonate)s (4 and 5) and poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s (6 and 7), were synthesized via an addition of poly(oxyethylene H-phosphonate)s to 9-anthrylidene-p-toluidine. The IR, NMR ((1)H, (13)C and (31)P) and fluorescence emission spectral data of the polymers are presented. The copolymers 6 and 7 were tested for in vitro antitumor activity on a panel of seven human epithelial cancer cell lines. Safety testing was performed both in vitro (3T3 NRU test) and in vivo on ICR mice for genotoxicity and antiproliferative activity. The copolymer 7 showed excellent antiproliferative activity to HBL-100, MDA-MB-231, MCF-7 and HepG2 cell lines. However, the in vitro safety testing revealed significant toxicity to Balb/c 3T3 mouse embryo cells. In contrast, the copolymer 6 showed complete absence of cytotoxicity to Balb/c 3T3 cells, but inhibited the growth of breast cancer cells, cervical carcinoma cells (HeLa) and hepatocellular carcinoma cell cultures after prolonged (72h) exposure. The polymers (4-6) exhibited low (4 and 6) to moderate (5) clastogenicity in vivo and slightly inhibited bone marrow cell division, compared to Mitomycin C. The subcellular distribution of the copolymers 6 and 7 were studied in model cell culture systems. The tested polyphosphoesters are expected to act in vivo as prodrugs of aminophosphonates and could be valuable as a new class of biodegradable polymer drug carriers.

Anthracene-Derived Bis-Aminophosphonates: Crystal Structure, In Vitro Antitumor Activity, and Genotoxicity In Vivo

Abstract The X-ray crystal structures of the anthracene-derived bis-aminophosphonates 4.4′-bis[N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]diaminodiphenylmethane (1) and 1,3-bis [N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]diaminobenzene (3) are reported. The X-ray analyses demonstrated that both compounds crystallize in a centrosymmetric manner containing a meso-form (1) and a pair of enantiomers (3). The cytotoxic potential, genotoxicity, and antiproliferative activity of bis-aminophosphonates 1 and bis[N-methyl(diethoxyphosphonyl)-1-(9-anthryl)]benzidine (2), as well as their subcellular distribution in a tumor cell culture system, are also discussed. Compounds 1 and 2 showed optimal antiproliferative activity to human tumor cells from colon carcinoma line HT-29. In vitro and in vivo safety testing revealed that the compounds exert lower toxicity to normal cells as compared with well-known anticancer and cytotoxic agents. Supplemental materials are available for this article. Go to the publishers online edition ofPhosphorus, Sulfur, and Silicon and the Related Elementsto view the free supplemental file. GRAPHICAL ABSTRACT

Low cytotoxicity and clastogenicity of some polymeric aminophosphonate derivatives

Poly(oxyethylene aminophosphonate)s synthesized on the basis of biodegradable poly(phosphorester)s and Schiff bases were tested in vitro for antitumor activity against a panel of six human epithelial cancer cell lines, for cytotoxicity to mouse fibroblast cells and in vivo for clastogenicity and antiproliferative effects. The polymers showed lower cytotoxicity, both in vivo and in vitro and lower clastogenicity in vivo than the corresponding low-molecular aminophosphonates. The biological activities of the tested polymers correlate with their low in vitro antitumor activity.

In vitro antitumour activity, genotoxicity, and antiproliferative effects of aminophosphonic acid diesters and their synthetic precursors.

The Schiff bases N-furfurylidene-p-toluidine and N-(4-dimethylaminobenzilidene)- p-toluidine, and the recently synthesized aminophosphonic acid diesters p-[N-methyl- (diethoxyphosphonyl)-(2-furyl)]toluidine and p-[N-methyl(diethoxyphosphonyl)-(4-dimethylaminophenyl)] toluidine were tested for in vitro antitumour activity on six human epithelial cancer cell lines. The genotoxicity and antiproliferative activity of these compounds were tested in mice. The aminophosphonates showed high in vitro antitumour activity towards the breast cancer-derived cell lines (MCF-7 and MDA-MB-231), the cervical carcinoma cell line (HeLa), and the human colon adenocarcinoma cell line (HT-29). In addition, the Schiff base N-furfurylidene-p-toluidine significantly inhibited the growth of bladder carcinoma cells (647-V) and the hepatocellular carcinoma line HepG2, and U-shaped dose-response curves were observed after treatment of 647-V and MCF-7 cells. All studied compounds had a moderate genotoxic and antiproliferative activity in vivo

In vitro antitumour activity, safety testing and subcellular distribution of two poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s in Ehrlich ascites carcinoma and BALB/c 3T3 cell culture systems

ABSTRACT Two polyphosphoesters containing anthracene-derived aminophosphonate and hydrophilic H-phosphonate repeating units, poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s (1 and 2), were tested for the in vitro antitumour activity on cell cultures derived from ascitic form of Ehrlich mammary adenocarcinoma by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-dye reduction assay. The in vitro safety testing of the copolymers was performed by BALB/c 3T3 neutral red uptake assay. A study on their uptake and subcellular distribution in non-tumourigenic and tumour cells was performed by means of fluorescence microscopy. Both copolymers showed significant antitumour activity towards Ehrlich ascites carcinoma (EAC) cells. However, the in vitro safety testing revealed significant toxicity of polymer 2 to BALB/c 3T3 mouse embryo cells. In contrast, polymer 1 showed complete absence of cytotoxicity to BALB/c 3T3 cells. The fluorescent studies showed that the substances were diffusely distributed in the cytoplasm in both cell culture systems. As opposed to BALB/c 3T3 cells, in EAC cells, intense fluorescent signal was observed in the nuclei and in the perinuclear region. The tested polyphosphoesters are expected to act under physiological conditions as prodrugs of aminophosphonates.

NOVEL MODELS OF AVIAN LEUCOSIS VIRUS-INDUCED CARCINOGENESIS

The LSCC-SF-MC29 cell culture model system was further characterized by studies on the provirus content of the cells, the host range and the subgroup specificity of the produced virus. The transforming potential of the Mc29 virus was evaluated by the focus-forming and colony-forming assays on primary cell cultures and continuous cell lines of avian and mammalian origin. The in ovo effects of the myelocytomatosis virus Mc29 on 15I line White Leghorn chicken embryos were studied by routine histopathological methods. Six avian leucosis virus-specific proviral sequences were detected by PCR analysis in the genome of LSCC-SF-MC29 cells. The presence of a Mc29 provirus-specific sequence located in the gag-myc region was confirmed. Using primers designed to differentiate ALV subgroups, amplification product was obtained with subgroup B/D-specific PCR primers. As it was expected, the subgroup E-specific PCR primers amplified the endogenous ALV sequences. In vitro studies on the host range of Mc29 virus showed that the primary cultures of chicken and hamster cells and a continuous hamster cell line were susceptible, while the cultures of primary quail cells and of a permanent line of duck embryo cells were resistant to the transforming effect of the virus. In ovo, the inoculated Mc29 virus induced hyperplasic and preneoplastic lesions in the embryonal liver and pancreas and myxomas of the neck.