Paridis saponins inhibiting carcinoma growth and metastasis <Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">in vivo</Emphasis>

Paris polyphylla Smith var. yunnanensis extracts, Rhizoma Paridis saponins (RPS) have been found to show strong antitumor activity. However, few studies have yet investigated pulmonary metastasis treatment with this herb. To detail the effective components in RPS and discuss the preliminary mechanism of antitumor effects in vivo and in vitro, a mixture isolated from RPS was investigated. The main constituents were identified as polyphyllin D, formosanin C, dioscin, Paris H, Paris VII and pennogennin 3-O-α-L-rhamnopyranosyl (1→4)-[β-L-rhamnopyranosyl (1→2)]-β-D-glucopyranoside. In our experiments, LA795 cells were exposed to the mixed compounds. Migration inhibition was evaluated by wound healing assay and migration assay in non-cytotoxic dose which was determined by MTT assay. The results demonstrated that the constituent in varying degrees inhibited the migration of the tumor cells in vitro. The mixture also showed antitumor effects on carcinoma in vivo. In conclusion, the mixture is a potent anticancer agent that elicits programmed cell death and inhibits the migration in murine lung adenocarcinoma, both in vitro and in vivo.     

Enhanced Intracellular Uptake of Sterically Stabilized Liposomal Doxorubicin <Emphasis Type="Italic">in Vitro</Emphasis> Resulting in Improved Antitumor Activity <Emphasis Type="Italic">in Vivo</Emphasis>

Purpose To investigate the correlation between the in vitro intracellular uptake and the in vivo antitumor activity of anticancer drugs delivered by sterically stabilized liposomes (SSL).Methods Arginine-glycine-aspartic acid (RGD) peptide or RGD mimetic (RGDm) was coupled onto the surface of SSL to obtain the cell-binding carrier to facilitate the intracellular delivery of the encapsulated drugs. DOX-loaded SSL (SSL-DOX), DOX-loaded RGD-modified SSL (RGD-SSL-DOX) and DOX-loaded RGDm-modified SSL (RGDm-SSL-DOX) were prepared by lipid film dispersion followed by remote loading of DOX. The intracellular uptake of DOX from the various liposomal formulations was evaluated in vitro with melanoma B16 cells, and the pharmacokinetics, biodistribution, and antitumor activity were compared in C57BL/6 mice carrying melanoma B16 tumors.Results In vitro intracellular uptake of DOX by B16 cells and in vivo antitumor activity in terms of tumor growth inhibition and mice survival time prolongation for various liposomal DOX were in the following order: RGD-SSL-DOX > RGDm-SSL-DOX > SSL-DOX. The mean survival time of the mice treated with RGD-SSL-DOX, RGDm-SSL-DOX, and SSL-DOX was 55, 49, and 44 days, respectively. The three liposomal DOX formulations produced very close DOX accumulation in tumor, which is significantly higher than that of free DOX. RGD- or RGDm-SSL-DOX demonstrated prolonged circulation time similar to that of SSL-DOX, whereas they showed significantly lower DOX level in blood and remarkably higher uptake by spleen than SSL-DOX.Conclusions Enhanced intracellular uptake of DOX encapsulated in SSL could produce an improved therapeutic effect for the melanoma B16 tumors. Enhancing intracellular delivery of the anticancer drugs encapsulated in SSL may be a promising strategy to improve their therapeutic efficacy for solid tumors.     

Stereoselective Taurine Conjugation of (<Emphasis Type="Italic">R</Emphasis>)-Benoxaprofen Enantiomer in Rats: <Emphasis Type="Italic">In Vivo</Emphasis> and <Emphasis Type="Italic">in Vitro</Emphasis> Studies Using Rat Hepatic Mitochondria and Microsomes

No HeadingPurpose. Identify (R)-BOP-T in rat bile after administration of (R)-BOT over a 12 h period.Methods. Each benoxaprofen (BOP) enantiomer was administered i.v. to bile duct-cannulated rats at a dose of 5 mg/kg. The optical isomers of BOP and its metabolites in plasma, urine, and bile were quantified using a chiral HPLC column. The amounts of BOP glucuronide (BOP-G), BOP taurine conjugate (BOP-T), and BOP enantiomers excreted into the bile over 12 h after administration of (R)-BOP were as follows: (R)-BOP-G and (S)-BOP-G, 2.1 ± 0.5 and 6.2 ± 1.4% of the dose; (R)-BOP-T and (S)-BOP-T, 5.6 ± 1.8 and 0.7 ± 0.3% of the dose; (R)-BOP and (S)-BOP, 0.7 ± 0.1 and 1.7 ± 0.2% of the dose, respectively, whereas after (S)-BOP administration, (S)-BOP-G and (S)-BOP were mainly excreted into the bile (14.3 ± 1.8 and 3.0 ± 0.4% of the dose, respectively). Only after (R)-BOP administration was the taurine conjugate of BOP found in the bile, and the configuration was R. BOP-T could not be found in the bile after (S)-BOP administration. To investigate the stereoselectivity of the conjugation enzymes responsible for BOP-T formation, in vitro studies were performed using rat hepatic organelles.Results. When (R)-BOP was used as a substrate, rat hepatic mitochondrial and microsomal fractions exhibited stereoselective BOP-T formation activity, with microsomal activity approximately 3.0 times greater than that of the mitochondria. That of (S)-BOP was approximately 2.1. Mean (R)/(S) ratios of BOP enantiomer for BOP-T formation in the mitochondrial and microsomal incubations were approximately 1.7 and 2.4, respectively.Conclusion. Although in the in vivo studies, only (R)-BOP-T originated from (R)-BOP was found in the bile, the configuration of BOP-T formed by the incubations of (R)-BOP or (S)-BOP with rat hepatic mitochondria or microsomes was S for both.     

Identification of medicinal <Emphasis Type="Italic">Dendrobium</Emphasis> species by phylogenetic analyses using <Emphasis Type="Italic">matK</Emphasis> and <Emphasis Type="Italic">rbcL</Emphasis> sequences

Species identification of five Dendrobium plants was conducted using phylogenetic analysis and the validity of the method was verified. Some Dendrobium plants (Orchidaceae) have been used as herbal medicines but the difficulty in identifying their botanical origin by traditional methods prevented their full modern utilization. Based on the emerging field of molecular systematics as a powerful classification tool, a phylogenetic analysis was conducted using sequences of two plastid genes, the maturase-coding gene (matK) and the large subunit of ribulose 1,5-bisphosphate carboxylase-coding gene (rbcL), as DNA barcodes for species identification of Dendrobium plants. We investigated five medicinal Dendrobium species, Dendrobium fimbriatum, D. moniliforme, D. nobile, D. pulchellum, and D. tosaense. The phylogenetic trees constructed from matK data successfully distinguished each species from each other. On the other hand, rbcL, as a single-locus barcode, offered less species discriminating power than matK, possibly due to its being present with little variation. When results using matK sequences of D. officinale that was deposited in the DNA database were combined, D. officinale and D. tosaense showed a close genetic relationship, which brought us closer to resolving the question of their taxonomic identity. Identification of the plant source as well as the uniformity of the chemical components is critical for the quality control of herbal medicines and it is important that the processed materials be validated. The methods presented here could be applied to the analysis of processed Dendrobium plants and be a promising tool for the identification of botanical origins of crude drugs.     

<Emphasis Type="Italic">Pulsatilla</Emphasis> saponin D: The antitumor principle from<Emphasis Type="Italic">Pulsatilla koreana</Emphasis>

By bioassay-guided separation, an already known saponin, Pulsatilla saponin D was isolated from the root of Pulsatilla koreana Nakai as a antitumor component when evaluated by in vivo antitumor activity as well as in vitro cytotoxic activity test. It showed potent inhibition rate of tumor growth (IR, 82%) at the dose of 6.4 mg/kg on the BDF1 mice bearing LLC cells.     

<Emphasis Type="Italic">In vivo</Emphasis> evidence of the immunomodulatory activity of orally administered <Emphasis Type="Italic">Aloe vera</Emphasis> gel

The gels of Aloe species contain immunomodulatory components such as aloctin A and acemannan. Most studies on these gels were performed in in vitro cell culture systems. Although several studies examined their immunomodulatory activity in vivo, the route of administration was intraperitoneal or intramuscular. Here, we evaluated the in vivo immunomodulatory activity of processed Aloe vera gel (PAG) in mice. Oral administration of PAG significantly reduced the growth of C. albicans in the spleen and kidney following intravenous injection of C. albicans in normal mice. PAG administration also reduced the growth of C. albicans in streptozotocin-induced diabetic mice. PAG administration did not increase ovalbumin (OVA)-specific cytotoxic T lymphocyte (CTL) generation in normal mice, but did increase it in high-fat-diet induced diabetic mice. These findings provide the first clear evidence for the immunomodulatory activity of orally administered Aloe vera gel.     

<Emphasis Type="Italic">In Vitro</Emphasis>/<Emphasis Type="Italic">in Vivo</Emphasis> Correlation of Transdermal Naltrexone Prodrugs in Hairless Guinea Pigs

Purpose The purpose of this investigation was to evaluate the in vitro and in vivo percutaneous absorption of the following prodrugs of naltrexone (NTX): 2-ethylbutyryl-3-O-ester-NTX (ETBUT-ester), methyl-3-O-carbonate-NTX (ME-carbonate), ethyl-3-O-carbamate-NTX (ET-carbamate), and N,N-dimethyl-3-O-carbamate-NTX (DME-carbamate) in hairless guinea pigs.Methods In vitro fluxes of NTX and its prodrugs through guinea pig skin were determined using a flow-through diffusion cell system. The pharmacokinetics of NTX prodrugs were determined after topical application of transdermal patches in guinea pigs.Results All the prodrugs hydrolyzed to NTX on passing through the skin, and ME-carbonate provided the highest NTX flux and had the highest apparent permeability coefficient (K_p). ME-carbonate and ET-carbamate underwent the highest extent of bioconversion to NTX upon passing through the skin as compared to ETBUT-ester and DME-carbamate. The results of the in vivo studies indicated that a significant amount of NTX was delivered after the application of transdermal patches of NTX prodrugs. A mean steady-state plasma concentration of 7.1 ng/ml was obtained after the application of transdermal patches of ME-carbonate. A good correlation was obtained between the in vitro and in vivo results.Conclusions The results of the in vivo studies indicated that the ME-carbonate prodrug of NTX was the most promising drug candidate for transdermal delivery.     

Synthesis and <Emphasis Type="Italic">in Vitro</Emphasis> cytotoxic activities of 2-alkyl-2,3-dihydro-1<Emphasis Type="Italic">H</Emphasis>-2,6-diazacyclopenta[<Emphasis Type="Italic">b</Emphasis>]anthracene-5,10-diones

A series of 2-alkyl-2,3-dihydro-1H-2,6-diazacyclopenta[b]anthracene-5,10-diones (4a–f) was synthesized and their in vitro cytotoxic activities were evaluated against six human cancer cell lines (HCT15, SK-OV-3, SNB19, A549, MCF7 and MCF7/ADR). They all appeared to be less potent than doxorubicin against all doxorubicin sensitive human cancer cell lines tested. However, these compounds retained considerable cytotoxic activity against the doxorubicin-resistant cell line MCF7/ADR, implying their therapeutic potential to treat doxorubicin-resistant tumors. The most active compound 4c was equipotent with doxorubicin against HCT15 cell line.     

Cross-linked Small Polyethylenimines: While Still Nontoxic,Deliver DNA Efficiently to Mammalian Cells <Emphasis Type="Italic">in Vitro</Emphasis> and <Emphasis Type="Italic">in Vivo</Emphasis>

No HeadingPurpose. Polyethylenimine (PEI) is among the most efficient nonviral gene delivery vectors. Its efficiency and cytotoxicity depend on molecular weight, with the 25-kDa PEI being most efficient but cytotoxic. Smaller PEIs are noncytotoxic but less efficient. Enhancement in gene delivery efficiency with minimal cytotoxicity by cross-linking of small PEIs via potentially biodegradable linkages was explored herein. The hypothesis was that cross-linking would raise the polycations effective molecular weight and hence the transfection efficiency, while biodegradable linkages would undergo the intracellular breakdown after DNA delivery and hence not lead to cytotoxicity. Toward this goal, we carried out cross-linking of branched 2-kDa PEI and its 1:1 (w/w) mixture with a linear 423-Da PEI via ester- and/or amide-bearing linkages; the in vitro and in vivo gene delivery efficiency, as well as toxicity to mammalian cells, of the resultant cross-linked polycations were investigated.Methods. The efficiency of the cross-linked PEIs in delivering in vitro a plasmid containing -galactosidase gene and their cytotoxicity were investigated in monkey kidney cells (COS-7). Dynamic light scattering was used to compare the relative DNA condensation efficiency of the unmodified and cross-linked PEIs. In vivo gene delivery efficiency was evaluated by intratracheal delivery in mice of the complexes of a luciferase-encoding plasmid and the PEIs and estimating the luciferase expression in the lungs.Results. Cross-linking boosted the gene delivery efficiency of the small PEIs by 40- to 550-fold in vitro; the efficiency of the most potent conjugates even exceeded by an order of magnitude that of the branched 25-kDa PEI. Effective condensation of DNA was evident from the fact that the mean diameter of the complexes of the cross-linked PEIs was some 300 nm with a narrow size distribution, while the complexes of the unmodified small PEIs exhibited a mean size of >700 nm with a very broad size distribution. At concentrations where the 25-kDa PEI resulted in >95% cell death, the conjugates afforded nearly full cell viability. The cross-linked PEIs were 17 to 80 times m ore efficient than the unmodified ones in vivo; furthermore, their efficiencies were up to twice that of the 25-kDa PEI.Conclusions. Cross-linking of small PEIs with judiciously designed amide- and ester-bearing linkers boosts their gene delivery efficiency both in vitro and in vivo without increasing the cytotoxicity. The high efficiency is dependent on the nature of the linkages and the PEIs used.     

<Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">In Vivo</Emphasis> Anticancer Activity of a Novel Nano-sized Formulation Based on Self-assembling Polymers Against Pancreatic Cancer

Purpose  

To evaluate the in vitro and in vivo pancreatic anticancer activity of a nano-sized formulation based on novel polyallylamine grafted with 5% mole cholesteryl pendant groups (CH₅-PAA).     

Population <Emphasis Type="Italic">In Vitro</Emphasis>-<Emphasis Type="Italic">In Vivo</Emphasis> Correlation Model for Pramipexole Slow-Release Oral Formulations

Purpose  

To establish an in vitro-in vivo level A correlation (IVIVC) for pramipexole slow-release formulations.     

<Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">In Vivo</Emphasis> Effects of Water Extract of White Cocoa Tea (<Emphasis Type="Italic">Camellia ptilophylla</Emphasis>) Against Human Prostate Cancer

Purpose  

We evaluated the chemotherapeutic effect of water extract of white cocoa tea (WCTE) against human prostate cancer (PCa) in vitro and in vivo.     

Nidurufin as a new cell cycle inhibitor from marine-derived fungus <Emphasis Type="Italic">Penicillium flavidorsum</Emphasis> SHK1-27

A new cell cycle inhibitor, nidurufin (1), was isolated from the marine-derived fungus Penicillium flavidorsum SHK1-27. An evaluation of antitumor activity indicated that 1 induced in vitro cell cycle arrest at G₂/M transition in the K562 cell line in a concentration and timedependent manner, with an IC₅₀ value of 12.6 M.     

<Emphasis Type="Italic">ent</Emphasis>-Abietane diterpenoids from <Emphasis Type="Italic">Isodon xerophilus</Emphasis>

Three new ent-abietanoids, named xerophilusins XIV–XVI, and four known analogues, as well as four known chemical constituents were isolated from the leaves of Isodon xerophilus. Their structures were elucidated by extensive spectroscopic studies, and comparison with literature data. In addition, the cytotoxic activity of the ent-abietanoids against chronic myelogenous leukemia (K562), stomach adenocarcinoma (MKN45), and hepatocellular carcinoma (HepG2) human cell lines was investigated and no activities were observed.     

Evaluation of <Emphasis Type="Italic">Sesbania grandiflora</Emphasis> for antiurolithiatic and antioxidant properties

In the indigenous system of medicine in India, the plant Sesbania grandiflora is claimed to be useful for various ailments, and one such use is for the treatment of renal calculi. The major purpose of this study is to investigate the potential of S. grandiflora in the treatment of renal calculi. The leaf juice of S. grandiflora was evaluated for median lethal dose, gross behavioral changes, antiurolithiatic and antioxidant activities. The antiurolithiatic activity was evaluated by a calculi-producing diet model, using gentamicin (subcutaneously) and 5% ammonium oxalate in rat feed to induce calcium oxalate-type stones. The parameters monitored in the present study are calcium and oxalate deposition in the kidney, kidney weights, urinary excretion of calcium and oxalate. The in vivo antioxidant parameters lipid peroxidation, glutathione reductase and catalase were monitored. The plant juice was also evaluated for scavenging of nitric oxide and 2-diphenyl-2-picryl hydrazyl free radicals. The leaf juice of S. grandiflora was safe orally and exhibited no gross behavioral changes except for an increase in urination. The leaf juice of S. grandiflora showed significant antiurolithiatic activity against calcium oxalate-type stones and also exhibited antioxidant properties. The results obtained in this study provide evidence for the efficacy of the leaf juice of S. grandiflora as antiurolithiatic agent.     

Pharmacological and histological effects of <Emphasis Type="Italic">Centaurea bruguierana</Emphasis> ssp. <Emphasis Type="Italic">belangerana</Emphasis> on indomethacin-induced peptic ulcer in rats

The species Centaurea bruguierana (DC.) Hand.-Mazz. ssp. belangerana (DC.) Bornm. (CBB) (Asteraceae), known as “Baad-Avard” in Borazjan, Bushehr Province, southern Iran, is used in folk medicine as a hypoglycemic herb in diabetes and as a remedy for peptic ulcer disorders. Total 80% EtOH extract and petroleum ether, CHCl₃, EtOAc, n-BuOH, and remaining fractions obtained by solvent–solvent fractionation of dried aerial flowering parts of the plant were investigated for anti-ulcer activity against indomethacin-induced ulcerogenesis in rats. Anti-ulcer activity was evaluated by measuring the ulcer index (UI) and ulcer inhibition. The UI was significantly reduced in all treated animals. A dramatic decrease in the UI was observed following the administration of total extract (100 mg/kg, p < 0.001) and CHCl₃ fraction (42 mg/kg, ***p < 0.001) in comparison with the control group. The percentage ulcer inhibition with total extract at a dose of 100 mg/kg (97.66%) and CHCl₃ fraction at a dose of 42 mg/kg (96.96%) was found to be higher (p < 0.001) than the reference group (cimetidine 100 mg/kg) (87.08%). The pharmacological and histological results of the present study proved that the aerial flowering parts of CBB possess preventive activity against peptic ulcer, supporting the traditional assertion in southern Iranian folk medicine.     

A Semi-mechanistic Modeling Strategy to Link <Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">In Vivo</Emphasis> Drug Release for Modified Release Formulations

Purpose  

To develop a semi-mechanistic model linking in vitro to in vivo drug release.     

Identification of <Emphasis Type="Italic">Curcuma</Emphasis> plants and curcumin content level by DNA polymorphisms in the <Emphasis Type="Italic">trn</Emphasis>S-<Emphasis Type="Italic">trn</Emphasis>fM intergenic spacer in chloroplast DNA

With the goal of developing an accurate plant identification method, molecular analysis based on polymorphisms of the nucleotide sequence of chloroplast DNA (cpDNA) was performed in order to distinguish four Curcuma species: C. longa, C. aromatica, C. zedoaria, and C. xanthorrhiza. Nineteen regions of cpDNA were amplified successfully via polymerase chain reaction (PCR) using total DNA of all Curcuma plants. Using the intergenic spacer between trnS and trnfM (trnSfM), all four Curcuma plant species were correctly identified. In addition, the number of AT repeats in the trnSfM region was predictive of the curcumin content in the rhizome of C. longa.     

In vitro cytotoxic activity of tri-<Emphasis Type="Italic">n</Emphasis>-butyltin(IV)lupinylsulfide hydrogen fumarate (IST-FS 35) and preliminary antitumor activity in vivo

Summary  The cytotoxicity in vitro and antitumor activity in vivo of the organotin compound tri-n-butyltin(IV)lupinylsulfide hydrogen fumarate (IST-FS 35) have been investigated. The IC50 values obtained in a panel of tumor cell lines were compared to those of the parental compound IST-FS 29 in the same cells. IST-FS 35 resulted significantly more active than IST-FS 29 with IC50 values in the range 0.16–1.8 μM. Toxicity studies in vivo, after intravenous administration of escalating concentrations of IST-FS 35, provided the identification of the maximal tolerated dose (3.5 mg/kg) which was employed as therapeutic dose in the antitumor activity experiments. Preliminary results, in transplanted murine tumor models, revealed that both the P388 myelomonocytic leukaemia and the B16-F10 melanoma, implanted subcutaneously in BDF1 mice, were inhibited about 96% in their tumor volume at day 11, following a single intravenous injection of the compound. Additional studies are mandatory to unravel the mechanism of action for the development of IST-FS 35 as potential antitumor drug.     

The synthesis and preliminary activity assay <Emphasis Type="Italic">In Vitro</Emphasis> of peptide-like derivatives as APN inhibitors

Both the aminopeptidase N (APN) and matrix metalloproteinase (MMP) are essential metallopeptidases in the development of tumor invasion and angiogenesis. A series of novel peptide-like derivatives were designed and synthesized as antitumor agents. Their structures were confirmed by IR, MS, and ¹H-NMR. These compounds exhibited potent inhibitory activities against APN and low activity against MMP in vitro. The derivatives with methoxy group show better activities than those with other substituted group and could be used as lead compounds for exploring new APN inhibitors in the future.