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Pharmaceutical cocrystals are an efficient approach to improve the solubility of insoluble active pharmaceutical ingredients (APIs), while the dissolution profiles of pharmaceutical cocrystals usually exhibit a type of “spring and rapid parachute” effect that influences stability and pharmacodynamic sustainability, in which the high apparent solubility induced by the formation of cocrystals can only be maintained for a short time (usually minutes) in the metastable zone and then decreases rapidly afterwords. Herein we reported the preparation and structures of two lenalidomide cocrystals with gallic acid (1, 2). After the formation of cocrystals, the intrinsic dissolution rate and apparent solubility of lenalidomide were found to be enhanced. The high solubility of cocrystals can keep for 48 h without dropping. The result of phase solubility study indicates gallic acid (GA) forms a 1:1 complex with lenalidomide (Rev) in aqueous solution, with a stability constant of 1713.2 L/mol. The multiple hydrogen bon...

An anticancer drug, lenalidomide, has low oral bioavailability (below 33%) due to its poor solubility in water. To improve its solubility, three cocrystals of lenalidomide with urea (1) and 3,5-dihydroxybenzoic acid (2, 3) were prepared. The structures of 1–3 were determined by single crystal X-ray diffraction, and they all form three-dimensional hydrogen-bonded frameworks between lenalidomide and the coformers. The apparent solubility value and intrinsic dissolution rate of lenalidomide in phosphate buffer of pH 6.8 have been improved after the formation of cocrystals. In addition, 2 and 3 can convert to each other under certain conditions.

Sperm protein 17 (Sp17) is selectively overexpressed in several human malignancies including ovarian carcinoma, but is absent or expressed at low levels in most normal tissues. Previous work from our group characterized an anti-Sp17 monoclonal antibody (clone 3C12) and showed that it specifically targeted tumor cells. In this report, we investigated whether a novel immunoconjugate containing 3C12 linked to the chemotherapeutic agent doxorubicin [(DOX) Adriamycin] had antitumor activity against ovarian cancer cell lines and tumor models. DOX was conjugated to 3C12 using a linker, and the specificity of 3C12-DOX was examined in Sp17-positive SKOV3 and Sp17-negative COC2 ovarian cancer cells using cell-based ELISA and internalization assays. The cytotoxicity of 3C12-DOX was assessed with the MTT assay, and its therapeutic effectiveness was evaluated in immunodeficient mice bearing SKOV3 cells. In vitro, the 3C12-DOX immunoconjugate specifically bound to and was internalized by Sp17-positive SKOV3 cells but did not bind to Sp17-negative cells. Treatment with 3C12-DOX (0.001 to 10 μg/mL) decreased the viability of SKOV3 cells in a Sp17-specific manner. In vivo, 3C12-DOX (3 mg/kg) induced the regression of established SKOV3 xenograft tumors in BALB/c mice compared with control treatment. The antitumor effects of 3C12-DOX were significantly associated with the induction of apoptosis in tumor cells. In addition, 3C12-DOX showed no observable adverse effects or toxicity when compared with DOX alone in mice bearing ovarian tumor xenografts. Our findings suggest that 3C12-DOX may be a potential antibody-drug conjugate for clinical development.

Aim To investigate the anti-inflammatory activity and possible mechanism of diosgenin, and to provide some further pharmacological evidence for its clinical applications. Methods Anti-inflammatory activities of diosgenin were evaluated by zymosan A-evoked peritoneal leukocytes migration in mice and the adhesion of human pro-myelocytic leukemia cell strain (HL-60) to human umbilical vein endothelial cell line (ECV304) cells induced by tumor necrosis factor (TNF-α1;). Furthermore, the effects of diosgenin on TNF-α1;-induced intercellular adhesion molecule-1 (ICAM-1) expression were also investigated by reverse transcription-PCR (RT-PCR) and flow cytometric analysis, and nuclear factor-kappaB (NF-αB)/p65 translocation and phosphorylation were determined by Western blot. Results Diosgenin significantly inhibited peritoneal leukocytes migration induced by zymosan A in mice when given orally once at doses of 1 and 3 mg·kg−1. Pretreatment ECV304 cells with diosgenin at concentrations of 0.1 and 1 μmol·L−1 for 5 h remarkably decreased TNF-α1;-stimulated adhesion of HL-60 to ECV304 cells in vitro, with no effect on viability of ECV304 cells. Furthermore, diosgenin inhibited TNF-α-induced overexpression of ICAM-1 both at the mRNA and protein levels, and suppressed nuclear p65 accumulation and p65 activation in ECV304 cells. Conclusion These results suggested that diosgenin significantly inhibited leukocytes migration and adhesion, partly due to the down-regulation of ICAM-1 expression through NF-κB pathway.

Sperm protein 17 (Sp17) is a cancer testis antigen that has been shown to be overexpressed in a variety of gynecologic malignancies, in particular ovarian cancer. Emerging evidences indicate that Sp17 is involved in tumorigenesis and in the migration of malignant cells. It has been proposed as a useful target for tumor-vaccine strategies and a novel marker to define tumor subsets and predict drug response. However, the antitumor activity of anti-Sp17 monoclonal antibody (anti-Sp17 mAb) has not been investigated. In this study, the in vitro cytotoxicity, antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) activities of anti-Sp17 mAb were evaluated using Sp17-positive ovarian cancer cells as targets, Sp17-negative ovarian cancer cells as the control, and healthy human peripheral blood monocytes and healthy human serum as effectors. Our preliminary results indicate that the direct cytotoxicity of anti-Sp17 mAb against the investigated ovarian cancer cells was very weak. However, the cytotoxicity of anti-Sp17 mAb, mediated by peripheral blood mononuclear cells (PBMCs), as ADCC, or by human serum, as CDC, was relatively strong in the Sp17-positive ovarian cancer cells. This finding suggested that anti-Sp17 mAb could be a useful tool against ovarian cancer and may provide insight into the development of low side-effect targeting therapy for this malignant disease.

Our previous results suggested that ruscogenin inhibited tumor necrosis factorα (TNF-α)–induced leukocyte adhesion, which correlated with its suppression of intercellular adhesion molecule-1 (ICAM-1) expression in endothelial cells. In the present studies, we further examined its effects on the main signaling pathways involved in upregulation of ICAM-1 induced by TNF-α in human umbilical vein endothelial cells (HUVECs). The results showed that ruscogenin significantly suppressed p65 phosphorylation, IκB-α phosphorylation and degradation, and inhibited IκB kinaseα (IKKα) and IKKβ activation induced by TNF-α. However, it exerted weak effects on TNF-α–induced phosphorylations of p38, JNK, ERK1/2, and Akt. Overall, our results indicated that downregulation of ICAM-1 expression by ruscogenin in HUVECs might be mediated by nuclear factor-κB (NF-κB), but not by mitogen-activated protein kinase (MAPK) and Akt signaling pathways. Keywords:: ruscogenin, nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK)

To construct the expression vector of small hairpin RNA (shRNA) and to investigate its efficacy in silencing nonmuscle myosin heavy chain IIA (MYH9)gene.According to the MYH9 cDNA sequence in GenBank, three shRNA expression vectors targeting MYH9 gene were constructed and were confirmed by digestion with restriction enzymes and DNA sequencing. The recombinant plasmids were transfected into HUVEC cells. The expression of MYH9 was determined by semi-quantitative RT-PCR at mRNA level and by western blotting at protein level at 24 h, 48 h and 72 h after transfection.The decrease of MYH9 expression at mRNA and protein levels gradually became more evident from 24 h to 48 h, and achieved the maximal degree at 48 h, then became weakened and restored at 72 h. The efficiency of pGCsi-MYH9-3 was the best among these three plasmids. The introduction of pGCsi-MYH9-3 was showed to efficiently and specifically inhibit the expression of MYH9 with inhibitory rate at 71.2% according to results of Western blot. RT-PCR results showed that mRNA transcription of MYH9 gene was reduced by 86.5% at 48 h after introduction of pGCsi-MYH9-3.Plasmid pGCsi-MYH9-3 could inhibit MYH9 expression in HUVEC cells successfully and effectively, which promised its further application in related function analysis of MYH9.

Ruscogenin (RUS), first isolated from Ruscus aculeatus, also a major steroidal sapogenin of traditional Chinese herb Radix Ophiopogon japonicus, has been found to exert significant anti-inflammatory and anti-thrombotic activities. Our previous studies suggested that ruscogenin remarkably inhibited adhesion of leukocytes to a human umbilical vein endothelial cell line (ECV304) injured by tumor necrosis factor-Α (TNF-Α) in a concentration-dependent manner. Yet the underlying mechanisms remain unclear. In this study, the in vivo effects of ruscogenin on leukocyte migration and celiac prostaglandin E2 (PGE2) level induced by zymosan A were studied in mice. Furthermore, the effects of ruscogenin on TNF-Α-induced intercellular adhesion molecule-1 (ICAM-1) expression and nuclear factor-κB (NF-κB) activation were also investigated under consideration of their key roles in leukocyte recruitment. The results showed that ruscogenin significantly suppressed zymosan A-evoked peritoneal total leukocyte migration in mice in a dose-dependent manner, while it had no obvious effect on PGE2 content in peritoneal exudant. Ruscogenin also inhibited TNF-Α-induced over expression of ICAM-1 both at the mRNA and protein levels and suppressed NF-κB activation considerably by decreasing NF-κB p65 translocation and DNA binding activity. These findings provide some new insights that may explain the possible molecular mechanism of ruscogenin and Radix Ophiopogon japonicus for the inhibition of endothelial responses to cytokines during inflammatory and vascular disorders. Keywords:: ruscogenin, endothelial cell, adhesion, intercellular adhesion molecule-1 (ICAM-1), nuclear factor-κB (NF-κB)