betadex and Carcinoma--Lewis-Lung

This work presents a very new look at folate targeting and is focused on synthesizing and assessing the biological activity of folic acid-targeted drug delivery materials based on β-cyclodextrin. Both folic acid and β-cyclodextrin have been covalently conjugated to branched polyethylenimine as the polymeric vector. Host-guest inclusion of folic acid into a β-cyclodextrin cavity, demonstrated by means of the spectroscopic methods (2-D NMR, IR, UV-Vis), is found to be of crucial importance for biological activity of nanotherapeutics. This paper describes the very first example of the versatile synthetic approach to create the polymeric biosystems, where folic acid activity is not limited by the inclusion phenomenon. Cytotoxicity of the obtained polymeric materials against Lewis lung carcinoma cells is determined by neutral red uptake assay. Folate receptor-binding studies reveal that the developed synthetic approach enables full exploitation of the potential of folic acid as a targeting ligand.

Topics: Animals; beta-Cyclodextrins; Carcinoma, Lewis Lung; Cell Survival; Folate Receptors, GPI-Anchored; Folic Acid; Magnetic Resonance Spectroscopy; Mice; Polyethyleneimine; Spectrophotometry, Ultraviolet

To analyze mechanisms for cancer metastasis, we established high metastatic sublines from mouse Lewis lung cancer (P29) by repeated injection. Sublines established from the two subclones H7 and C4 commonly exhibited increased proliferation and invasion activity and reduced expression of ganglioside GM1, although they showed different preferences in their target organs of metastasis. The high metastatic sublines secreted higher levels of activated matrix metalloprotease (MMP)-9 as well as pro-MMP-9 in the culture medium than the parent lines. Furthermore, they contained MMP-9 at the glycolipid-enriched microdomain (GEM)/rafts fractionated by the sucrose density gradient ultracentrifugation of Triton X-100 extracts, whereas the parent cells showed faint bands at the fraction. When high metastatic sublines were treated with methyl-beta-cyclodextrin, their invasion activities were dramatically suppressed, and the MMP-9 secretion was also suppressed. All these results indicated that GEM/rafts play crucial roles in the increased invasion and high metastatic potential. To clarify the implication of reduced GM1 expression, low GM1-expressing cell lines were established using an RNA interference-expression vector of the GM1 synthase. Low GM1-expressing cell lines showed increased proliferation and invasion, enrichment in the GEM/rafts, and increased secretion of MMP-9. Among adhesion molecules, only integrin beta1 was detected in GEM/rafts with stronger intensity in high metastatic lines and low GM1-expressing cells. Taken together, integrins seemed to be enriched in the GEM/rafts by reduced GM1 levels, and subsequently MMP-9 was recruited to the GEM/rafts, resulting in its efficient secretion and activation, and eventually in the increased invasion and metastatic potentials.

Topics: Animals; beta-Cyclodextrins; Carcinoma, Lewis Lung; Cell Line, Tumor; Down-Regulation; G(M1) Ganglioside; Immunohistochemistry; Integrins; Lung Neoplasms; Matrix Metalloproteinase 9; Membrane Microdomains; Membrane Proteins; Mice; Mice, Inbred C57BL; Neoplasm Invasiveness; Neoplasm Transplantation; RNA, Small Interfering; Transfection

The effect of hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) on the aqueous solubility and chemical stability of O-(4-Dimethylaminoethoxycinnamoyl)fumagillol (CKD-732), a new angiogenesis inhibitor, was investigated with an aim of preparing a stable and effective parenteral formulation. The CKD-732/HP-beta-CyD inclusion complex was obtained in solid state by freeze-drying and characterized in solution by proton nuclear magnetic resonance (1H NMR). Then, the pharmacokinetic profile in rats and the in vivo tumor growth inhibitory activity in mice following the parenteral administration of aqueous CKD-732/HP-beta-CyD complex were compared to those of CKD-732.hemioxalate solution having an equivalent concentration. The aqueous solubility of CKD-732 was markedly increased by the combination of pH adjustment and HP-beta-CyD complexation through a soluble 1:1 inclusion complex formation, which was supported by NMR spectroscopy. The hydrolysis of CKD-732 following pseudo first-order kinetics was decelerated moderately but significantly in acidic and basic solutions in the presence of HP-beta-CyD. The stability of lyophilized CKD-732/HP-beta-CyD complex was also drastically improved after storage in various conditions. The intravenous pharmacokinetic profile and the subcutaneous in vivo tumor growth inhibitory activity of aqueous CKD-732/HP-beta-CyD complex were not significantly different from those of CKD-732.hemioxalate solution with the favorable reduction of irritation. These results demonstrate that the CKD-732/HP-beta-CyD complex is an attractive formulation for use in the parenteral delivery of CKD-732.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Angiogenesis Inhibitors; Animals; beta-Cyclodextrins; Carcinoma, Lewis Lung; Cinnamates; Cyclodextrins; Cyclohexanes; Drug Compounding; Drug Stability; Drug Storage; Epoxy Compounds; Hydrolysis; Injections, Intravenous; Mice; Pharmaceutical Solutions; Phase Transition; Rats; Rats, Sprague-Dawley; Sesquiterpenes; Solubility; Time Factors