Potential Use of Folate-polyethylene glycol (PEG)-Appended Dendrimer (G3) Conjugate with alpha-Cyclodextrin as DNA Carriers to Tumor Cells

We previously reported that polyamidoamine STARBURST dendrimer (generation 3, G3) (dendrimer) conjugate with alpha-cyclodextrin (alpha-CyD) having an average degree of substitution of 2.4 of alpha-CyD (alpha-CDE) provided remarkable aspects as novel carriers for DNA and siRNA. To develop novel alpha-CDE derivatives with tumor cell specificity, we prepared folate-appended alpha-CDEs (Fol-alpha-CDEs) and folate-polyethylene glycol (PEG)-appended alpha-CDEs (Fol-PalphaCs) with the various degrees of substitution of folate (DSF), and evaluated in vitro and in vivo gene transfer activity, cytotoxicity, cellular association and physicochemical properties. In vitro gene transfer activity of Fol-alpha-CDEs (G3, DSF 2, 5 or 7) was lower than that of α-CDE (G3) in KB cells, folate receptor (FR)-overexpressing cancer cells. Of the three Fol-PalphaCs (G3, DSF 2, 5 or 7), Fol-PalphaC (G3, DSF 5) had the highest gene transfer activity in KB cells. The activity of Fol-PalphaC (G3, DSF 5) was significantly higher than that of alpha-CDE (G3) in KB cells, but not in A549 cells, FR-negative cells. Negligible cytotoxicity of the pDNA complex with Fol-PalphaC (G3, DSF 5) was observed in KB cells or A549 cells up to a charge ratio of 100/1 (carrier/pDNA). The cellular association of the pDNA complex with Fol-PalphaC (G3, DSF 5) could be mediated by FR on KB cells, resulting in its efficient cellular uptake. Fol-PalphaC (G3, DSF 5) had higher binding affinity with folate binding protein (FBP) than alpha-CDE (G3), although the physicochemical properties of pDNA complex with Fol-PalphaC (G3, DSF 5) were almost comparable to that with alpha-CDE (G3), although the onset charge ratio and the compaction ability of Fol-PalphaC (G3, DSF 5) were slightly different. Fol-PalphaC (G3, DSF 5) tended to show higher gene transfer activity than alpha-CDE (G3) 12 h after intratumoral administration in mice. These results suggest that Fol-PalphaC (G3, DSF 5), not Fol-alpha-CDEs, could be potentially used as a FR-overexpressing cancer cell-selective DNA carrier

Synthesis and Isolation of Regiospecific Mono Iodo-Cyclodextrin as a Fragile Intermediate to Amino-Cyclodextrin

lodo-CD (II) was prepared from toluenesulfonyl-CD (I) and purified by preparative HPLC. Iodo-CD (II) was easily converted to the amino-CD (III). Examination of the structure of the hydrolyzate from amino-CD (III) showed that indination and amination proceeded with inversion of configuration in both reactions

The Pavlik harness in the treatment of developmentally dislocated hips: results of Japanese multicenter studies in 1994 and 2008

AbstractBackgroundIt has already been more than 50years since the Pavlik harness was introduced in Japan, and today the Pavlik harness is widely recognized as the standard initial treatment modality for developmental dysplasia of the hip. We performed a multicenter nationwide questionnaire study concerning the results of Pavlik harness treatment twice in 1994 and 2008.MethodsIn 1994 and in 2008, we sent questionnaires to 12 institutes in Japan specializing mainly in pediatric orthopedics. We compare the results of these two studies and discuss differences in reduction rates, incidence of avascular necrosis in the femoral epiphysis and the percentage of joints with acceptable morphology (Severin grade I+II/total) at skeletal maturity. We statistically assessed these results to see whether there were changes in the treatment outcomes over this 14-year period.ResultsReduction of the dislocated hips was obtained by the Pavlik harness in 80.2% (1990/2481 hips; 1994) and 81.9% (1248/1523 hips; 2008). The incidences of avascular necrosis of the proximal femoral epiphysis in the dysplastic hips were 14.3% (119/835 hips; 1994) and 11.5% (76/663 hips; 2008). The type of avascular necrosis in hips from the 2008 study was determined according to the classification of Kalamchi and MacEwen: 24/69 hips (34.8%) were classified as group I; 20/69 hips (29.0%) as group II; 11/69 hips (15.9%) as group Ill; 14/69 hips (20.3%) as group IV. The percentages of hips with acceptable outcomes at skeletal maturity discerned from Severin X-ray changes (grade I+II/total) were 72.3% (604/835 hips; 1994) and 77.7% (488/628 hips; 2008).ConclusionReduction rates and the incidence of avascular necrosis in 2008 were statistically similar to the results in 1994. The rate of acceptable outcome (Severin grade I+II/total) in 2008 was statistically higher than that of 1994

Hydrolytic Catalysis with Complexation by Modified Cyclodextrins Having Diastereomeric Structure

At the presence of three regiospecifically modified cyclodextrins, i. e., β-CD-D-histidine (I_D), β-CD-L-histidine (I_L) and β-CD-histamine (III), the ester cleavage at neutral pH was compared concerning in enantiomeric selectivity as well as rate enhancement. Micha-elis-Menten-type kinetics were observed for I_D and I_L in the phenol release of acetylanyl (and phenylalanyl) p-nitrophenyl ester enantiomers. Second order rate constants were obtained for III. Stereochemical comparisons of the spatial geometry in the complexes were discussed and also the schematic scheme of the host-guest fit was discussed

The Effects of Cyclodextrins on the Reduction of Ninhydrin with N-Alky1-1, 4-dihydronicotinamides

The effects of adding various cyclodextrins (α-,β- and γ-cyclodextrin) on the redox reaction between N-alky1-1, 4-dihydronicotinamide (hydride ion donnor) and ninhydrin (hydride ion acceptor) in aqueous solutions were studied kinetically. These reactions proceed through the hydride ion donnor-acceptor complex (CT complex) formation without cyclodextrin (CD). The presence of CD especially affects the reduction with N-benzy1-1,4-dihydronicotinamide (BNAH). β-CD is most effective for decreasing the reduction rate in three CDs. β-CD inhibited the formation of the CT complex by including BNAH molecule, which caused a decrease in the reaction rate. β-CD also completely inhibited the hydration of BNAH. γ-CD also included BNAH, however, it had little effect on the reduction rate constant. α-CD had no effect on both the reduction and the hydration. These inhibition behaviors were caused by an inclusion complex formation with CD

Chiral Recognition in the Inclusion Complex of Cyclodextrins with N-Dansyl-Phenylalanine Observed by Linked Scan FAB-MS Spectrometry

The cyclodextrin (CD) inclusion complex showed a chiral recognition observed by linked scan FAB-MS. The ratio, R, of the daughter ionic peak strength to the precursor ionic peak strength using the linked scan FAB-MS method was suggested to express a parameter for the stability of CD inclusion complex. The ratios between R_D and R_L were determined in inclusion complexes consisted of D-and L-N-dansyl-phenylalanines with various CDs. As for this parameter R derived from the linked scan FAB-MS ionic peaks, the wide application is possible to the inclusion complexes and the evaluation of the chiral recognition in various inclusion complexes

ケイコウプローブ ニ ヨル メチルカシクロデキストリン ノ ホウセツケイタイ ノ ケントウ

The fluorescence spectra of 8-anilino-1-naphthalene sulfonic acid (ANS) and 7-toluidino-2-naphthalene sulfonic acid (TNS) were observed with or without various cyclodextrins. From the change of fluorescence intensity, the association constants between cyclodextrins and fluorescent probes (K) and the fluorescence intensity constants (i_0) for each cyclodextrin complex were calculated. The value of i_0 depended on the probe\u27s local microenvironment. These parameters assist in explication of the complex formation. And the effect of methylation of cvclodextrin for inclusion behavior is also indicated

The Effects of Methylated Cyclodextrins on the Transphosphorylation among AMP, ADP and ATP in the Presence of Escherichia Coli

Transphosphorylation, 2ADP⇄AMP+ATP, occurred in the presence of intact Escherichia coli (E. coli. strain K-12) in neutral phosphate buffer solution, because adenylate kinase existed in both cytoplasma and periplasma of E. coli. Cyclodextrin(CD)s accelerated the transphosphorylation. The reaction rate depended on the concentration of CD and the kind of CD. Methylated CDs were more effective than parent CD, especially, hiptakis-(2,6-di-O-methyl)-β-CD(DMCD) was most effective. Addition of CDs makes neither change of E. coli concentration nor the leak of adenylate kinase from periplasma of E. coli

N-ビオチニル-アミノ-β-シクロデキストリン ノ ゴウセイ

The fact that α-biotin serves as a covalently bound "C0_2 carrier" is now unequivocally established. Though model studies such as carboxylation on ureid l\u27-N-of biotin has been reported, no appropriate biomimetic model study for a "C0_2 carrier" has been carried out. The titled compound(I)in which the biotinyl group is bound to an amide linkage withβ-cyclodextrin, was prepared with active-ester method in good yield. Compound I was purified by taking with preparative HPLC. The structure of (I) was providedby ^1H-nmr, ^C-nmr, elemental analysis, HPLC and the paper choromatography of hydrolyzates, and decided to be 6-(mono-N-biotinyl)-amino-β-cyclodextrin