ABSTRACT
The environment-sensitive probe 2 with turn-on switch for Mcl-1 protein was developed herein. After careful evaluation, this small molecule fluorescent probe revealed a selective binding affinity with Mcl-1 protein with a Ki value of 2.6 µM and can be well applied to imaging Mcl-1 protein or detecting the cellular distribution of Mcl-1 protein inhibitors. Compared with other imaging approaches, such as the immunofluorescence and fluorescent protein-based techniques, this fluorescent method is rapid, convenient, and affordable, especially since a washing procedure is no longer needed. This environment-sensitive "off-on" design strategy may present a case in point for developing small-molecule fluorescent probe of Bcl-2 family proteins.
Subject(s)
Dansyl Compounds/chemistry , Fluorescent Dyes/chemistry , Indoles/chemistry , Myeloid Cell Leukemia Sequence 1 Protein/analysis , Cell Line, Tumor , Dansyl Compounds/chemical synthesis , Dansyl Compounds/toxicity , Fluorescent Dyes/chemical synthesis , Fluorescent Dyes/toxicity , HEK293 Cells , Humans , Indoles/chemical synthesis , Indoles/toxicity , Microscopy, Confocal/methods , Microscopy, Fluorescence/methods , Molecular Docking Simulation , Myeloid Cell Leukemia Sequence 1 Protein/chemistryABSTRACT
A novel amphiphilic imidazolium-based probe containing a dansyl fluorophore and a long cetyl chain has been developed for ATP recognition. The probe forms self-assembled micelle-like aggregates at low concentration in its aqueous solution and can selectively recognize ATP among other bioactive anions with a significant enhancement in fluorescence emission.
Subject(s)
Adenosine Triphosphate/analysis , Dansyl Compounds/chemistry , Fluorescent Dyes/chemistry , Imidazoles/chemistry , Surface-Active Agents/chemistry , Apyrase/analysis , Cell Membrane Permeability , Dansyl Compounds/chemical synthesis , Dansyl Compounds/toxicity , Enzyme Assays , Fluorescent Dyes/chemical synthesis , Fluorescent Dyes/toxicity , Hep G2 Cells , Humans , Hydrogen-Ion Concentration , Imidazoles/chemical synthesis , Imidazoles/toxicity , Surface-Active Agents/chemical synthesis , Surface-Active Agents/toxicityABSTRACT
PURPOSE: To investigate the use of nano self-assemblies formed by polyallylamine (PAA) modified with 5 or 10% mole fluorenylmethoxy carbonyl (Fmoc(5)/(10)), dimethylamino-1-naphthalenesulfonyl (Dansyl(5)/(10)) and 5% mole cholesteryl group (Ch(5)) for oral hydrophobic drug delivery. METHODS: Propofol, griseofulvin and prednisolone were loaded into amphiphilic PAAs. Particle size and morphology of drug-loaded self-assemblies were determined using photon correlation spectroscopy and transmission electron microscopy. Solubilising capacity, in vitro drug release and formulation stability were analysed by HPLC, and in vitro biocompatibility studies (haemolysis and cytotoxicity) were carried out on bovine erythrocytes and Caco-2 cells, respectively. Dansyl(10) and Ch(5) griseofulvin formulations were administered intra-gastrically to rats, and drug plasma levels were analysed by HPLC. RESULTS: Drug-encapsulated self-assemblies typically have hydrodynamic size of 300-400 nm. Dansyl(10) exhibited universal drug solubiliser property and had significantly improved prednisolone, griseofulvin and propofol solubility by 145, 557 and 224-fold, respectively. Fmoc polymers resulted in modest drug solubility improvement. These polymers were non-haemolytic, did not enhance cytotoxicity compared to unmodified PAA, and demonstrated significant increase in griseofulvin plasma concentration compared to griseofulvin in water after oral administration. CONCLUSIONS: Ch(5) and Dansyl(10) showed promising potential as nano-carriers for oral hydrophobic drug delivery.
Subject(s)
Anti-Inflammatory Agents/administration & dosage , Drug Carriers/chemistry , Griseofulvin/administration & dosage , Hypnotics and Sedatives/administration & dosage , Polyamines/chemistry , Prednisolone/administration & dosage , Propofol/administration & dosage , Administration, Oral , Animals , Anti-Inflammatory Agents/pharmacokinetics , Caco-2 Cells , Cattle , Cell Survival , Cholestenes/chemistry , Cholestenes/toxicity , Dansyl Compounds/chemistry , Dansyl Compounds/toxicity , Drug Carriers/toxicity , Fluorenes/chemistry , Fluorenes/toxicity , Griseofulvin/pharmacokinetics , Hemolysis/drug effects , Humans , Hydrophobic and Hydrophilic Interactions , Hypnotics and Sedatives/pharmacokinetics , Male , Polyamines/toxicity , Prednisolone/pharmacokinetics , Propofol/pharmacokinetics , Rats , Rats, Sprague-DawleyABSTRACT
Fluorescent dyes (e.g., dansyl, fluoresceine isothiocyanate, or naphthalimide groups) are widely used as markers to study biological properties of drugs. In order to evaluate possible mediated cytotoxicity, we attached three molecules each to 1,3,5-tris(3-propylamino)benzene initially synthesized as core molecule for the design of dendrimers. Cytotoxic effects were only observed for the NO(2)-substituted naphthalimide conjugate. The intracellular distribution was visualized via confocal fluorescence microscopy and pointed to an accumulation in the endosome or nucleus, dependent on the cell line used.
