Hydrophobic polyamine analogs and methods for their use
a polyamine analog and hydrophobic technology, applied in the field of chemistry and biochemistry, can solve the problems of limited value of use in the design of mammalian transport inhibitors, and achieve the effect of suppressing atrial tachycardia and inhibiting polyamine transpor
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example i
Chemical Synthesis of Polyamine Agents (PAs)
[0131]PAs analogs were synthesized in a parallel fashion starting from the orthogonally protected diamino containing amino acid starting materials. The use of the 4-nitrophenyl activated ester L-Boc-Lys-(Cbz)-ONP in FIG. 1 provides an exemplary illustration of the synthetic process. The active ester is added dropwise to a solution of 1.5 equivalents of polyamine in methanol to give a statistical mixture of unsubstituted, mono-substituted and di-substituted acyl polyamines. Following evaporation of the solvent, the remaining free amino groups in the polyamine moiety are protected either as their tBoc or Cbz carbamates. Standard workup results in a completely protected crude product mixture. The desired orthogonally-protected product is isolated in pure form by silica gel chromatography using standard organic solvents. This purification process is based on separation of polyamine molecules with the remaining amino groups being fully protecte...
example ii
Cell Culture and Reagents
[0134]All cell lines were obtained from ATCC (Manassas, Va.) and cultured in the recommended media, serum, and CO2 concentration. Medias were obtained from Mediatech, Inc. (Herdon, Va.) and serums from Gibco BRL (Gaithersburg, Md.). 50 U / ml penicillin, 50 μg / ml streptomycin and 2 mM L-glutamine (all from Bio Whittaker, Walkersville, Md.) were included in all cultures. DFMO was obtained from Marion Merrell Dow (Cinncinati, Ohio). When cells were cultured with polyamines or ORI compounds, 1 mM aminoguanidine (AG; Sigma) was included to inhibit serum amine oxidase activity. IC50 refers to the concentration of PA that results in 50% of maximum cell growth inhibition in the presence of PA alone.
example iii
Polyamine Transport and Ki Assays
[0135][2,9-3H]spermidine (SPD) from DuPont NEN, Boston, Mass. was added alone or simultaneously with PAs to 24-well plates containing MDA-MB-23 1 cells in log growth. The cells were incubated at 37° C. for 15 min to determine initial rate polyamine uptake. The cells were then washed three times with cold PBS, lysed with 0.1% SDS, and the amount of polyamine incorporation into the cells was determined by scintillation counting of the cell lysates. To determine a Ki, four radioactive substrate concentrations (0.3-3 μM) and five inhibitor concentrations (0.01-1.0 μM) and a control were tested. The Ki values were determined using double reciprocal Lineweaver-Burke plot analyses. Ki values were determined from linear equations derived from graphing the slopes of Lineweaver-Burke plot vs. inhibitor concentration, with Ki=y-intercept / slope. Results of these analyses are shown in Table 3 above.
PUM
| Property | Measurement | Unit |
|---|---|---|
| logP | aaaaa | aaaaa |
| logP coefficient | aaaaa | aaaaa |
| logP | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More