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[Display omitted] Oligonucleotide therapeutics, particularly antisense oligonucleotides (ASOs), have emerged as promising candidates in drug discovery. However, their effective delivery to the target tissues and cells remains a challenge, necessitating the development of suitable drug delivery technologies for ASOs to enable their practical application. In this study, we synthesized a library of chemically modified dipeptide-ASO conjugates using a recent synthetic method based on the Ugi reaction. We then conducted in vitro screening of this library using luciferase-expressing cell lines to identify ligands capable of enhancing ASO activity. Our findings suggest that N -(4-nitrophenoxycarbonyl)glycine may interact with the thiophosphate moiety of the phosphorothioate-modification in ASO. Through our screening efforts, we identified two ligands that modestly reduced luciferase luminescence in a cell type-selective manner. Furthermore, quantification of luciferase mRNA levels revealed that one of these promising dipeptide-ASO conjugates markedly suppressed luciferase RNA levels through its antisense effect in prostate-derived DU-145 cells compared to the ASOs without ligand modification. [ABSTRACT FROM AUTHOR]

Oligonucleotide therapeutics are generally prepared via solid-phase synthesis, and many impurities derived from the synthesis process are present in crude oligonucleotides. Among the various impurities typical of oligonucleotide synthesis, shortmers, which lack a few residues from the desired oligonucleotide therapeutics, are difficult to remove using conventional high-performance liquid chromatography (HPLC) purification because they have similar structures and physical properties to the desired oligonucleotide. In this study, we developed a simple purification method for oligonucleotide therapeutics. In our strategy, the retention of shortmers on the reversed-phase HPLC column was increased by using a highly lipophilic phosphoramidite, which is a reagent for 5′-end modification of oligonucleotides, instead of acetic anhydride during solid-phase synthesis, and only the target oligonucleotide could be eluted from a reversed-phase HPLC pretreatment column. [Display omitted] [ABSTRACT FROM AUTHOR]

3′,4′-Ethyleneoxy-bridged 5-methyluridine derivatives with methyl groups in the bridge, ( R )-Me-3′,4′-EoNA-T and ( S )-Me-3′,4′-EoNA-T, were synthesized, and these two analogs and unsubstituted 3′,4′-EoNA-T were successfully incorporated into a 2′,5′-linked oligonucleotide ( iso DNA). Their duplex-forming ability with complementary DNA and complementary RNA, and triplex-forming ability with double-stranded DNA, were evaluated by UV-melting experiments. The results indicated that iso DNAs, including these 3′,4′-EoNA analogs, could hybridize exclusively with complementary RNA. In particular, 3′,4′-EoNA-T and ( R )-Me-3′,4′-EoNA-T modifications within iso DNA could stabilize the duplexes with complementary RNA compared with unmodified or 3′,4′-BNA-modified iso DNAs. [ABSTRACT FROM AUTHOR]

Few clinical studies have determined the quantitative transfer of vaginal chloramphenicol to circulating blood in pregnant women. This study aimed to evaluate the plasma concentration of chloramphenicol in pregnant women treated with trans-vaginal tablets and its relationship with maternal background and neonatal health. Thirty-seven pregnant women treated with 100 mg of trans-vaginal chloramphenicol once daily for bacterial vaginosis and its suspected case were enrolled. The plasma concentration of chloramphenicol was determined using liquid chromatography coupled to tandem mass spectrometry at day 2 or later after starting the medication. The correlations between the maternal plasma concentration of chloramphenicol and the background and neonatal health at birth were investigated. Chloramphenicol was detected from all maternal plasma specimens and its concentration ranged from 0.043 to 73.1 ng/mL. The plasma concentration of chloramphenicol declined significantly with the administration period. The plasma concentration of chloramphenicol was lower at the second than the first blood sampling. No correlations were observed between the maternal plasma concentration of chloramphenicol and background such as number of previous births, gestational age at dosing, and clinical laboratory data. Neonatal infant health parameters such as birth-weight, Apgar score at birth, and gestational age at the time of childbearing were not related to the maternal plasma concentration of chloramphenicol. Vaginal chloramphenicol transfers to circulating blood in pregnant women. The maternal plasma concentration of chloramphenicol varied markedly and was associated with the administration day, but not with maternal background or her neonatal health. [ABSTRACT FROM AUTHOR]

Background Few reports in the literature relate morphologic changes of the ulnar collateral ligament (UCL) to prior elbow symptoms. This study used ultrasonography (US) to assess the ulnohumeral joint space width, with and without stress, and elucidate morphologic changes of the UCL of the elbow in high school pitchers with and without a history of elbow symptoms. Methods Each of 122 high school baseball pitchers who underwent US of the medial aspect of both elbows completed a self-administered questionnaire related to the self-satisfaction score (0-100) for pitching performance and throwing-related elbow joint pain sustained during the prior 3 years. We conducted gravity stress US elbow examination with 30° of flexion with and without valgus stress. Comparisons of the UCL thickness and ulnohumeral joint space width, with and without valgus stress, were made among the 122 high school pitchers with and without a history of elbow symptoms. Results Pitchers with an elbow symptom history exhibited a greater difference between the UCL thickness on the throwing side than those with no elbow symptom history ( P  = .0013). A negative significant association was found between UCL thickness on the pitching side and the self-evaluation score for pitching performance ( r  = −0.20, P  = .04). Conclusions US assessment demonstrated that the UCL in the dominant side with elbow symptom history was thicker than that with no elbow symptom history. The UCL thickness might reflect the prior pitching condition of high school baseball pitchers. [ABSTRACT FROM AUTHOR]

Objectives Pharmacokinetic disposition of bortezomib in the blood has not been fully characterized in humans. This study aimed to evaluate the blood distribution of bortezomib and its kinetics in multiple myeloma patients. Design and method Eighteen multiple myeloma patients receiving bortezomib–dexamethasone combination therapy were enrolled. Blood specimens were drawn just before bortezomib administration on days 1 and 8 in the second and third cycles and after discontinuation. The relationships between bortezomib concentration and blood components were evaluated. Results Bortezomib concentration in the blood on day 1 was higher than that on day 8 in the second cycle. No difference was observed in bortezomib blood concentrations between day 8 in the second and third cycles. The bortezomib concentration in the blood and blood cells was 3- and 7-fold higher than that in plasma. Bortezomib concentration in the blood was correlated with the red blood cell count. The half-life of bortezomib in the blood was 23 days. Conclusion Bortezomib was taken up into red blood cells to only a limited extent and eliminated in parallel to the red blood cells' lifespan. The turnover of red blood cells can affect the pharmacokinetic disposition of bortezomib in multiple myeloma patients. [ABSTRACT FROM AUTHOR]

Abstract: Artificial thymidine monomers possessing amide or N-methylamide bridges were designed, synthesized, and introduced into oligonucleotides. UV-melting experiments showed that these oligonucleotides preferred single-stranded RNA (ssRNA) to single-stranded DNA (ssDNA) in duplex formation. Both amide- and N-methylamide-modified oligonucleotides led to a significant increase in the binding affinity to ssRNA by up to +4.7 and +3.7°C of the T m value per modification, respectively, compared with natural oligonucleotide. In addition, their oligonucleotides showed high stability against 3′-exonuclease. [Copyright &y& Elsevier]

Thymidine derivatives bearing spiroacetal moieties on the C4ʹ-position (5ʹ R -spiro-thymidine and 5ʹ S -spiro-thymidine) were synthesized and incorporated into oligonucleotides. The duplex- and triplex-forming abilities of both the oligonucleotides were evaluated from UV melting experiments. Oligonucleotides with the 5ʹ S -spiro modifications could form thermally stable duplexes with complementary RNA and DNA; however, the 5ʹ R -spiro modification significantly decreased the thermal stabilities of the duplexes and triplexes. Oligonucleotides with these spiro-thymidines showed significantly high resistance towards enzymatic degradation. [ABSTRACT FROM AUTHOR]

To determine the usefulness of monkey as an animal model, bioavailability and in vivo release behaviors of theophylline (TP) after oral administration of controlled-release beads in dogs, monkeys, and minipigs were evaluated. Controlled-release beads were prepared using a centrifugal-fluid type granulator, that is, CF-granulator, and Ethylcellulose (EC) was used as controlled-release coating agent. Aqueous solution and EC-coated beads, which contained TP were orally administered to animals after at least 1-week intervals. In dogs and minipigs, their relative bioavailabilities of EC-coated beads were 33.1% and 47.0%, respectively, and in vivo TP release from EC-coated beads in the gastrointestinal tract of dogs and minipigs were not reflected in vitro data. In monkeys, relative bioavailability of EC-coated beads was 80.0% and the highest among the three species, and release amount of TP from EC-coated beads at 24 h after oral administration was 82.8% and 92.4%, which was almost correlated to in vitro data. Therefore, the discrepancy of the relative bioavailability in three species is considered to be due to the difference of in vivo release behavior of TP. The monkey may be useful animal model for bioavailability studies of controlled-release dosage forms of TP from the viewpoint of in vitro-in vivo release correlation. © 2006 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 95: 1888–1895, 2006 [ABSTRACT FROM AUTHOR]

Abstract A practical and operationally simple post-synthetic modification of oligonucleotides containing 5-trifluoromethylpyrimidine bases is described. Trifluoromethyl group was used as a post-synthetic precursor and 5-trifluoromethylpyrimidine bases within oligonucleotides were converted into the corresponding 5-carboxy-, 5-cyano-, 5-amidinyl-, and 5-carbamoyl derivatives by treatment with an alkaline solution and amines. Moreover, post-synthetic treatment of fully protected and controlled pore glass (CPG)-attached oligonucleotides proceeded successfully with the simultaneous removal of all protecting groups, cleavage from CPG, and conversion of the trifluoromethyl group to afford the corresponding modified oligonucleotides. Graphical abstract Image 1 [ABSTRACT FROM AUTHOR]

Ceramic sheet of Na + ion-conducting solid electrolyte, Na 3 Zr 2 Si 2 PO 12 (NASICON), has been prepared by tape-casting method. Addition of 60Na 2 O-10Nb 2 O 5 -30P 2 O 5 glass as a sintering aid is crucial for fabrication of fully-densified impermeable sheet. Otherwise the gas tightness is not achieved even sintered at 1200 °C. The best preparation condition is 5 or 10 wt% grass-addition with respect to NASICON and sintering at 1000–1100 °C, which is lower by 100–200 °C than the typical densification temperature of NASICON without the additive. A sample with a thickness of 35 μm and total conductivity of 4.4 × 10 −4  S cm −1 is obtained by 5 wt% glass addition and sintering at 1100 °C for 4 h. [ABSTRACT FROM AUTHOR]

[Display omitted] This study was aimed at developing a novel platform for tetravalent conjugation of 4-arm polyethylene glycol (PEG) with an antisense oligonucleotide (ASO). The ASO technology has several limitations, such as low cellular uptake, poor nuclease stability, and short half-life. PEG-conjugated ASOs may result in an improvement in the pharmacokinetic behavior of the drug. Moreover, PEGylation can reduce enzymatic degradation and renal excretion of the conjugates, thereby, increasing its blood stability and retention time. In this study, we successfully synthesized PEG-ASO conjugate consisting of 4-arm-PEG and four molecules of ASO (4-arm-PEG-tetra ASO). Its hybridization ability with complementary RNA, enzymatic stability, and in vitro gene silencing ability were evaluated. No significant difference in hybridization ability was observed between 4-arm-PEG-tetra ASO and the parent ASO. In addition, gene silencing activity of the 4-arm-PEG-tetra ASO was observed in vitro. However, the in vitro activity of the 4-arm-PEG-tetra ASO was slightly reduced as that of the parent ASO. Moreover, the 4-arm-PEG-tetra ASO showed appreciable stability in cellular extract, suggesting that it hybridizes with mRNA in its intact form, without being cleaved in the cell, and exhibits ASO activity. [ABSTRACT FROM AUTHOR]

[Display omitted] The artificial nucleobase 1,3-diaza-2-oxophenoxazine (tCO) and its derivative G-clamp strongly bind to guanine and, when incorporated into double-stranded DNA, significantly increase the stability of the latter. As the phenoxazine skeleton is a constituent of major pharmaceuticals, we hypothesized that oligonucleotides (ONs) containing phenoxazine bases would induce property changes related to intracellular uptake and migration in tissues. In this study, we designed and synthesized a novel G-clamp-linker antisense oligonucleotide (ASO) in which a G-clamp base with a flexible linker was introduced into the 5′-end of an ASO targeting mouse long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (m MALAT1). Compared to unconjugated ASO, the G-clamp-linker ASO induced significantly more effective knockdown of m MALAT1 in mouse skeletal muscle. The ASOs conjugated with 2′-deoxyribonucleotide(s) bearing a tCO nucleobase at the 5′-end exhibited a similar knockdown effect in skeletal muscle. Thus, it may be possible to improve therapeutic effects against skeletal muscle diseases, such as muscular dystrophy, by using ONs with incorporated phenoxazine nucleobases. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study is to design a new orally disintegrating tablet (ODT) containing micronized ethylcellulose (MEC). The new ODT was prepared by physical mixing of rapidly disintegrating granules (RDGs) with MEC. To obtain RDGs, mannitol was spray-coated with a suspension of corn starch and crospovidone (9:1, w/w ratio) using a fluidized-bed granulator (suspension spray-coating method). The new ODTs were evaluated for their hardness, friability, thickness, internal structure (X-ray-CT scanning), in vivo disintegration time, and water absorption rate. Since MEC increases tablet hardness by increasing the contact frequency between the granules, the new ODTs could obtain high hardness (>50N) and low friability (<0.5%) with relatively low compression force. In addition, fine capillary channels formed in ODTs facilitated the wicking action and enabled rapid disintegration in vivo (<30s). On the other hand, since MEC has low hygroscopicity, the tablet hardness of ODTs containing MEC remained high for 1 month in high-humidity conditions. In conclusion, the new ODTs containing MEC developed in this study possessed superior properties for clinical use and are expected to be applied for a wide range of functionally released drugs for bitter taste masking, sustained release, and controlled release (pH-dependent film coating, matrix, and microcapsule). [Copyright &y& Elsevier]