Therapeutic potential and pharmacological activities of Atractylodes lancea (Thunb.) DC.

The rhizome of Atractylodes Iancen(A.lancea)(Thunb.) DC.(AL)is extensively used in Chinese,Thai,and Japanese traditional medicines as crude extracts/decoctions or a component in various herbal formulations.Various pharmacological activities of Al.and its major constituents have been demonstrated in ritro.ex ciro.and in animal models.Results from the toxicity studies in animal models suggest safety profile of AL,and its active constituents.Despite extensive use with positive impression in many diseases,there has not been a clinical study that can conclusively support its efficacy and safely profile in human.This review comprehensively summarizes current information on the pharmacological activities of AL and their active constituents including anticancer,anti-inflammatory,antimicrobial and antipyretic activities,as well as activities on central nervous,cardiovascular,and gastrointestinal systems.

Polymorphisms of the oxidant enzymes glutathione S-transferase and glutathione reductase and their association with resistance of Plasmodium falciparum isolates to antimalarial drugs

Objective:To investigate the association between amplification of the two regulatory genes controlling glutathione(GSH) levels,glutathione reductase(PfGR) and glutathione S-transferase (PfGST) genes and sensitivity of Plasmodium falciparum(P.falciparum) isolates collected from different malaria endemic areas of Thailand to standard antimalarial drugs.Methods:A total of 70 P.falciparum isolates were collected from endemic areas of multi-drug resistance (Tak,Chantaburi and Ranong Provinces) during the year 2008-2009.The in vitro assessment of antimalarial activity of P.falciparum clones(K1- and Dd2 chloroquine resistant and 3D7- chloroquine sensitive) and isolates to chloroquine,quinine,mefloquine and arteusnate was performed based on SYBR Green modified assay.Results:68(97.14%),11(15.71%) and 28(40%) isolates respectively were classified as chloroquine-,quinine- and mefloquine-resistant isolates. With this limited number of P.falciparum isolates included in the analysis,no significant association between amplification of PfGST gene and sensitivity of the parasite to chloroquine, quinine,mefloquine and quinine was found.Based on PCR analysis,Dd2,Kl and 3D7 clones all contained only one copy of the PfGST gene.All isolates(70) also carried only one copy number of PfGST gene.There appears to be an association between amplification of PfGR gene and chloroquine resistance.The 3D7 and Dd2 clones were found to carry only one PfGR gene copy, whereas the K1 clone carried two gene copies.Conclusions:Chloroquine resistance is likely to be a consequence of multi-factors and enzymes in the GSH system may be partly involved. Larger number of parasite isolates are required to increase power of the hypothesis testing in order to confirm the involvement of both genes as well as other genes implicated in glutathione metabolism in conferring chloroquine resistance.

Antimalarial activity and toxicity of Garcinia mangostana Linn.

Objective:To investigate the antimalarial activity and toxicity of the crude ethanolic extract of its pericarp both in vitro and in vim.Methods:The antimalarial activity of Gareinja mangostana(G.mangostana)Linn.extract against 3D7 and Kl Plasmodium falciparum(P.falciparum)clone were assessed using SYBR green I-based assay.A 4-day suppressive test of Plasmodium berghei{P.berghei)infected mouse was performed to investigate in vivo antimalarial activity.Results:The in vitro antimalarial activity was seleclive(SI>5?and classified as weak and good lo moderate activity against both 3D7 and K1 P.falciparum,clones with median IC_(50)(range)values of 11.12(10.94-11.29)and 7.54(6.80-7.68)μg/mL,respectively.The extract was considered nontoxic to mice.The maximum tolerated doses for acute and subacute toxicity in mice were 5 000and 2 000 mg/kg,respectively.Median(range)parasite density on day 4 of the negative control group(25%Tween-80),mice treated with 250,500,1000,and 2 000 mg/kg body weight of the extract,and 10 mg/kg body weight of chloroquine for 14 d were 12.8(12.2-13.7),11.4(9.49-13.8),11.6(9.9-12.5),11.7(10.6-12.8),10.9(9.4-11.6)and 0(0-0)%respectively.Parasite density on day 4in the control group treated with Tween-80 was higher than the groups treated with chloroquine and all dose levels of the extract.Conclusions:G.mangostana linn,showed weak antimalarial activity of the extract both in vitro and in vivo could be due to limitation of absorption of the active compounds.

In vitro inhibitory effects of plumbagin,the promising antimalarial candidate,on human cytochrome P450 enzymes

Objective:To investigate the propensity of plumbagin to inhibit the three isoforms of human cytochrome P450(CYP),ie.,CYP1A2,CYP2C19,and CYP3A4 using human liver microsomes in ritro.Methods:Inhibitory effects of plumbagin on the three human CYP isoformswere investigated using pooled human liver microsomes.Phenacetin O-deethylation,omeprazole hydroxylation and nifedipine oxidation were used as selective substrates for CYP1A2,CYP2C19 and CYP3A4 activities,respectively.Concentrations of paracetamol,5-hydroxyomeprazole,and oxidized nifedipine were determined in microsomal incubation mixture using high performance liquid chromatography.Results:Plumbagin showed significantinhibitory effects on all CYP isoforms.but with the most potent activity on CYP2C19-mediated omeprazole hydroxylation.The IC50(concentration that inhibits enzyme activity by 50%) values of plumbagin and nootkatone(selective inhibitor) for CYP2C19 were(0.78±0.01) and(27.31±0.66) μM,respectively.The inhibitory activities on CYP1 A2-mediated phenacetin O-deethylation and CYP3A4-mediated nifedipine oxidation were moderate.The IC_(50) values of plumbagin and-naphthoflavone(selective inhibitor) for CYP1A2 were(1.39±0.01) and(0.02±.0.36) μM,respectively.The corresponding IC_(50) values of plumbagin and ketoconazole(selective inhibitor) for CYP3A4 were(2.37+0.10) and(0.18±0.06) μM,respectively.Conclusions:Clinical relevance of the interference of human drug metabolizing enzymes should be aware of for further development scheme of plumbagin as antimalarial drug when used in combination with other antimalarial drugs which are metabolized by these CYP isoforms.

Polymorphic patterns of pfcrt and pfmdrl in Plasmodium falciparum isolates along the Thai-Myanmar border

Objective:To investigate the distribution and patterns of pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum(P.falciparum)isolates collected from the malaria endemic area of Thailand along Thai-Myanmar border.Methods:Dried blood spot samples were collected from 172 falciparum malaria patients prior received treatment.The samples were extracted using chelex to obtain parasite DNA.PCR-RFLP was employed to detect pfert mutation at codons 76,220,271,326,3S6 and 371,and the pfmdr1mutation at codon 86.Pfmdr1 gene copy number was determined by SYBR Green 1 real-time PCR.Results:Mutant alleles of pfcrt and wild type allele of pfmdrl were found in almost all samples.Pfmdrl gene copy number in isolates collected from all areas ranged from 1.0 to S.0 copies and proportion of isolates carrying>1 gene copies was 38.1%.The distribution and patterns of pfcrt and pfmdrl mutations were similar in P.falciparum isolates from all areas.However,significant differences in both number of gfmdr1 copies and prevalence of isolates carrying>1 gene copies were observed among isolates collected from different areas.The median pfmdr1 copy number in P.falciparum collected from Kanchanaburi and Mae Hongson were 2.5 and 2.0,respectively and more than half of the isolates carried>1 gene copies.Conclusions:The observation of pfindr1 wild type and increasing of gene copy number may suggest declining of artesunate-mefloquine treatment efficacy in P.falciparum isolates in this border area.

Possible role of PGD_2 in malaria infections

Objective:To preliminarily investigate the possible role of prostaglandin D_2(PGD_2) in malaria infections.Methods:Blood and urinary samples(n=120 each) were collected from Thai patients with Plasmodium falciparum(P.falciparum) with moderate(n=26) and high(n=4) parasitemia,patients with Plasmodium vivax(P.vivax)(n=30),patients with fever associated with other infections(n=30),and healthy subjects(n=30).PGD_2 concentrations in plasma and urinary samples of healthy subjects,patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit(Cayman Chemical,USA).Results:The possible association between PGD_2 and malaria infections is clearly demonstrated with PGD_2 concentration in urine.The urinary PGD_2 concentrations were relatively high(about 5-fold) in patients with P.falciparum with moderate parasitemia and P.vivax infections compared with other groups.Furthermore,the concentration in patients with P.falciparum with moderate parasitemia and P.vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.Conclusions:Urinary PGD_2 concentrations may offer a more dependable and useful tool for predicting malaria severity.Confirmation is this preliminary finding is required with a larger sample size.

Glycoproteomics analysis of plasma proteins associated with Opisthorchis viverrini infection-induced cholangiocarcinoma in hamster model

Objective: To apply lectin affinity chromatography and glycoproteomics-based LC-MS/MS to preliminarily investigate the possible potential plasma biomarkers of Opisthorchis viverrini(OV)-associated CCA in OV/dimethylnitrosamine(DMN)-induced CCA hamster model. Methods: Nine Syrian hamsters were divided into 3 groups as follows(n=3 each): normal(healthy control group); OV group; and OV/DMN group(CCA group). Pooled plasma samples collected from animals in each group at the 6th month post-infection with OV metacercarae were subjected to glycoproteomics analysis. Glycoproteins in the pooled sample from each group were initially isolated by concanavilin A(Con A)-based affinity chromatography. The expression of glycoproteins isolated by both enrichment methods were determined using LCMS/MS. Results: Among the 24 Con A-binding glycoproteins isolated, two proteins, N-myc downstream regulated gene 1(NDRG1) and fetuin-B(FETUB) were found up-regulated only in the samples from the OV and control groups, but not in the OV/DMN(CCA) groups. On the other hand, one protein, i.e., NSFL1 cofactor p47 isoform x3(NSFL1C) was found only in the samples from OV/DMN(CCA) and control groups, but not in the OV group. The remaining 21 proteins were upregulated in the samples from all groups. Conclusions: NDRG1, FETUB and NSFL1 C glycoproteins isolated by Con A-based affinity chromatography could be potential biomarkers for CCA. Plasma samples with negative for NDRG1 and FETUB proteins but positive for NSFL1 C are likely to be OV-associated CCA. Nevertheless, this conclusion remains to be confirmed whether this battery test can discriminate OV-associated CCA from other risk factors.

Inhibitory activities of plumbagin on cell migration and invasion and inducing activity on cholangiocarcinoma cell apoptosis

Objective: To investigate the cytotoxic, apoptotic and inhibitory activities on cell migration and invasion of plumbagin in the human cholangiocarcinoma(CCA) cell line(CL-6) in comparison with human embryonic fibroblast cell line(OUMS). Methods: Cytotoxicity activity was evaluated using MTT assay. Inhibitory effect on cell migration and invasion were investigated using label-free real-time cell analysis and QCM ECMatrix cell invasion chamber, respectively. Apoptotic activity was evaluated using flow cytometry and Cell Event? Caspase 3/7 assay. Results: Based on results of the cytotoxicity test in CL-6 cells, 50% inhibitory concentration(IC_(50), Mean±SD) values of plumbagin and the standard drug 5-fluorouracil were(24.00±3.33) and(1 036.00±137.77) μmol/L, respectively. The corresponding values for OUMS cells were(57.00±5.23) and(2 147.00±209.98) μmol/L, respectively. The selectivity index was 2.28. The inhibitory activities of plumbagin on cell migration and invasion were potent and concentration-dependent with IC_(50) of 25.0 μmol/L and complete inhibition at 25.0 μmol/L. Flow cytometry analysis showed that plumbagin at 12.5 μmol/L(half IC_(50)) induced CL-6 cell apoptosis(43.24% of control) through stimulation of caspase 3/7 activities. Complete cell apoptosis was observed at 12.5 μmol/L. Conclusions: The cytotoxic activity and inhibition of migration and invasion including apoptosis induction in the human CCA cell line(CL-6) suggest that plumbagin could be a promising candidate for CCA chemotherapeutics. However, its relatively low selective cytotoxic effect on CCA cells is a major concern.

A brief review on biomarkers and proteomic approach for malaria research

Malaria remains as one of the significant health threat to people living in countries throughout tropical and subtropical zones.Proteomic studies of Plasmodium,the protozoan causing malaria,is essential for understanding its cellular structure,growth stage-specific expression of protein metabolites and complex interaction with host.In-depth knowledge of the pathogen is required for identification of novel biomarkers that can be utilized to develop diagnostic tests and therapeutic antimalarial drugs.The alarming rise in drug-resistant strains of Plasmodium has created an urgent need to identify new targets for drug development that can act by obstructing life cycle of this parasite.In the present review,we briefly discuss on role of various biomarkers including Plnsmodium-assaciated aldolase,histidine-rich proteins and lactate dehydrogenase for diagnosis of malaria.Here we also summarize the present and future prospects of currently used techniques in proteomic approaches such as two dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight(MALDI-TOF)for diagnosis and potential identification of novel proteins for malaria research.

Comparison of protein patterns between Plasmodium falciparum mutant clone T9/94-M1-1(b3) induced by pyrimethamine and the original parent clone T9/94

Objective:To compare the protein patterns from the extracts of the mutant clone T9/94-M1-1(b3)induced by pyrimethamine,and the original parent clone T9/94 following separation of parasite extracts by two-dimensional electrophoresis(2-DE).Methods:Proteins were solubilized and separated according to their charges and sizes.The separated protein spots were then detected by silver staining and analyzed for protein density by the powerful image analysis software.Results:Differentially expressed protein patterns(up—or down-regulation)were separated from the extracts from the two clones.A total of 223 and 134 protein spots were detected from the extracts of T9/94 and T9/94-M1-1(b3)clones,respectively.Marked reduction in density of protein expression was observed with the extract from the mutant(resistant)clone compared with the parent(sensitive)clone.A total of 25 protein spots showed at least two-fold difference in density,some of which exhibited as high as ten-fold difference.Conclusions:These proteins may be the molecular targets of resistance of Plasmodium falciparum to pyrimethamine.Further study to identify the chemical structures of these proteins by mass spectrometry is required.

The role of heme-oxygenase-1 in pathogenesis of cerebral malaria in the co-culture model of human brain microvascular endothelial cell and ITG Plasmodium falciparum-infected red blood cells

Objective: To investigate the role of human host heme-oxygenase-1(HO-1) in pathogenesis of cerebral malaria in the in vitro model,Methods: The effect of human host HO-1 [human brain microvascular endothelial cell(HBMEC)] on hemoglobin degradation in the co-culture model of HBMEC and ITG Plasmodium falciparum-infected red cells(i RBC) through measurement of the enzymatic products iron and bilirubin,Results: Following exposure to the HO-1 inducer Co PPIX at all concentrations,the HBMEC cells apoptosis occurred,which could be prominently observed at 15 μM of 3 h exposure,In contrast,there was no significant change in the morphology in the non-exposed i RBC at all concentrations and exposure time,This observation was in agreement with the levels of the enzymatic degradation products iron and bilirubin,of which the highest levels(106.03 and 1 753.54% of baseline level,respectively) were observed at 15 μM vs,20 μM at 3 h vs,24 h exposure,For the effect of the HO-1 inhibitor Zn PPIX,HBMEC cell morphology was mostly unchanged,but significant inhibitory effect on cell apoptosis was seen at 10 μM for the exposure period of 3 h(37.17% of baseline level),The degree of the inhibitory effect as reflected by the level of iron produced was not clearly observed(highest effect at 10 μM and 3 h exposure),Conclusions: Results provide at least in part,insight into the contribution of HO-1 on CM pathogenesis and need to be confirmed in animal model.

Proteomics analysis of antimalarial targets of Garcinia mangostana Linn.

Objective:To investigate possible protein targets for antimalarial activity of Garcina mangostana Linn.(G.mangostana)(pericarp)in 3D7 Plasmodium falciparum clone using 2-dimensional electrophoresis and liquid chromatography mass-spectrometry(LC/MS/MS).Methods:3D7 Plasmodium falciparum was exposed to the crude ethanolic extract of G.mangostana Linn.(pericarp)at the concentrations of 12μg/mL(1C_(50)level:concentration that inhibits parasite growth by 50%)and 30μg/mL(1C_(90)level:concentration that inhibits parasite growth by 90%)for 12 h.Parasite proteins were separated by 2-dimensional electrophoresis and identified by LC/MS/MS.Results:At the IC_(50)concentration,about 82%of the expressed parasite proteins were matched with the control(non-exposed),while at the IC_(90)concentration,only 15%matched proteins were found.The selected protein spots from parasite exposed to the plant extract at the concentration of 12μg/mL were identified as eneymes that play role in glycolysis pathway,i.e.,phosphoglyeerate mutase putative,L-lactate dehydrogenase/glyceraldehyde-3-phosphate dehydrogenase,and fruetose-bisphosphate aldolase/phosphoglyeerate kinase.The proteosome was found in parasite exposed to 30μg/mL of the extract.Conclusions:Results suggest that proteins involved in the glycolysis pathway may be the targets for antimalarial activity of G.mangostana Linn.(pericarp).

Atractylodin-loaded PLGA nanoparticles：formulation and characterization

OBJECTIVE To formulate atractylodin-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and characterize the prepared nanoparticle formulation.METHODS The nanoparticle formulation was developed using solvent displacement method.The encapsulation and loading efficiency were characterized and particle size,and zeta potential were determined by dynamic light scattering technique.Drug release was assessed in vitro.RESULTS The size(mean±SD of diameter) of the prepared atractylodin-loaded PLGA nanoparticles were(161.27 ± 1.87)nm with narrow size distribution(mean PDI:0.068±0.015) and zeta potential(28.83±0.35)mV.The encapsulation and loading efficiency were(48.31±0.83)% and(2.15±0.04)%,respectively.Drug release from atractylodin-loaded PLGA nanoparticles was observed up to(87.70±0.47)% in 72 h with biphasic manner.Moreover,the nanoparticles were found to be freely dispersible in water without aggregation.CONCLUSION Results suggest that PLGA nanoparticles may be used as an effective drug delivery system for atractylodin.The anti-cholangiocarcinoma activity of this nanoparticle formulation is required.

Identification of potential protein targets of atractylodin against cholangiocarcinoma

OBJECTIVE To identify potential cell signaling pathways and protein targets of the active compound isolated from Atracylodes lancea "atractylodin" in cholangiocarcinoma,using proteomics approach.METHODS The holangiocar-cinoma cell line was exposed with atractylodin for 3 and 6 h and the proteins from both intra-and extra-cellular components were extracted.The extract proteins were separated by SDS-PAGE and digested with trypsin.The LC-MS/MS was applied to identify proteins.Signaling pathways and protein expression were analyzed by MASCOT and STITCH software.RESULTS A total of 4,323 and 4,318 proteins were identified from intra-and extracellular components,respectively.Six intracellular proteins were linked with the signaling pathways(apoptosis,cell cycle control,and PI3K-AKT).Four extracellular proteins were linked with the signaling pathways(NF-κB and PI3K-AKT).CONCLUSION All these proteins will further study to confirm the link to the anticholangiocarcinoma ac.tivity of actractylodin.

Preliminary investigation on the prevalence of malaria and HIV co-infection in Mae Sot District, Tak Province of Thailand

Objective: To preliminarily investigate the prevalence of HIV co-infection in patients with malaria in Mae Sot District, Tak Province of Thailand.Methods: The study was a retrospective study on blood samples collected from a total of 256 patients with malaria(all species and severity) who attended Mae Tao clinic for migrant workers, Tak Province during 2005-2007(148 samples) and 2010-2012(108 samples). Malaria diagnosis was performed based on microscopic examination of patients' blood smears. Chemiluminescent microparticle immunoassay and gel particle passive agglutination were employed for the detection of HIV antigen in patients' plasma. Results: Plasmodium falciparum(P. falciparum) and Plasmodium vivax(P. vivax) are the two predominant malaria species with the ratio of about 1: 1 to 1.5:1. Most of the P. falciparum cases were presented with acute uncomplicated signs and symptoms with highest parasitemia of 1 045 000 asexual parasites/μL bloods. The prevalence of malaria and HIV co-infection during 2005-2007 was 1.35%(2/148 cases, 1 each for P. falciparum and P. vivax co-infection), but was increased to 2.78%(3/108 cases, 2 and 1 for P. falciparum and P. vivax co-infection, respectively) during 2010-2012.Conclusions: The increasing trend of prevalence of malaria and HIV co-infection in Mae Sot, Tak province was of a great concern on either pharmacodynamics or pharmacokinetics aspect. The study in a larger numbers of malaria patients in different endemic areas throughout the country with different time periods is underway.

Simulation of optimal dose regimens of photoactivated curcumin for antimicrobial resistance pneumonia in COVID-19 patients:A modeling approach

Background:Secondary antimicrobial resistance bacterial(AMR)pneumonia could lead to an increase in mortality in COVID-19 patients,particularly of geriatric patients with underlying diseases.The comedication of current medicines for AMR pneumonia with corticosteroids may lead to suboptimal treatment or toxicities due to drug-drug interactions(DDIs).Objective:This study aimed to propose new promising dosage regimens of photoactivated curcumin when co-administered with corticosteroids for the treatment of antimicrobial resistance(AMR)pneumonia in COVID-19 patients.Methods:A whole-body physiologically-based pharmacokinetic(PBPK)with the simplified lung compartments model was built and verified following standard model verification(absolute average-folding error or AAFEs).The pharmacokinetic properties of photo-activated were assumed to be similar to curcumin due to minor changes in physiochemical properties of compound by photoactivation.The acceptable AAFEs values were within 2-fold.The verified model was used to simulate new regimens for different formulations of photoactivated curcumin.Results:The AAFEs was 1.12-fold.Original formulation(120 mg once-daily dose)or new intramuscular nano-formulation(100 mg with a release rate of 10/h given every 7 days)is suitable for outpatients with MRSA pneumonia to improve patient adherence.New intravenous formulation(2000 mg twice-daily doses)is for hospitalized patients with both MRSA and VRSA pneumonia.Conclusion:The PBPK models,in conjunction with MIC and applied physiological changes in COVID-19 patients,is a potential tool to predict optimal dosage regimens of photo-activated curcumin for the treatment of co-infected AMR pneumonia in COVID-19 patients.Each formulation is appropriate for different patient conditions and pathogens.

In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component,ethyl-p-methoxycinnamate, against cholangiocarcinoma

Objective:To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG)extract and ethyl-p-methoxycinnamate(EPMC)in vitro.Methods:The present study investigated the cytotoxic[using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphe nyl-2-H-tetrazolium bromide test],apoptotic(using a mitochondrial membrane potential assay),mutagenic(using a micronucleus test)and immunomodulatory(using flow cytometry)activities of the ethanolic extract of KG and its bioactive component,EPMC,against two cholangiocarcinoma(CCA)cell lines,CL-6 and HuCCT1,and one normal human cell line,OUMS-36 T-1 F.Results:Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells.The cytotoxic activity was supported by their concentration-dependent induction of apoptosis.CL-6 was most sensitive(3–4 fold)and selective to 5-fluorouracil(5-FU),compared with KG extract and EPMC[median half inhibiting concentration(IC50)and selectivity index(SI)were 23.01 lg/mL and 17.32;78.41 lg/mL and 4.44;100.76 lg/mL and 2.20,respectively for 5-FU vs.KG extract vs.EPMC].HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract[median IC50 and SI were 66.03 lg/mL and6.04;60.90 lg/mL and 3.65;156.60 lg/mL and 2.23,respectively for 5-FU vs.EPMC vs.KG extract].EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells(IC50=92.20 lg/mL).KG extract and EPMC exhibited concentration-dependent mutagenic activity,as well as inhibition of tumor necrosis factor-a and interleukin-6.Conclusion:Considering cytotoxic,apoptotic,immunomodulatory and mutagenic activities,further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.

Development of oral pharmaceutical formulation of standardized crude ethanolic extract of Atractylodes lancea(Thunb) DC.

Cholangiocarcinoma(CCA), the adenocarcinoma of the biliary duct, is commonly reported in Asia with the highest incidence in northeastern Thailand. Chemotherapy of this type of cancer is limited due to the lack of effective chemotherapeutic drugs. A series of previous studies support further research and development of Atractylodes lancea(Thunb) DC.(AL) as a potential candidate for the treatment of CCA as a crude ethanolic extract. In the present study, we aimed to develop an oral pharmaceutical formulation(capsule) of the standardized AL crude ethanolic extract for further clinical development in patients with CCA. Major steps included macroscopic and microscopic authentication of the AL rhizomes, preparation of standardized AL extract, preparation of oral pharmaceutical formulation(capsule) of the standardized AL extract, quantitative and qualitative analysis of the marker compound(atractylodin) in the formulated AL extract, evaluation of contaminations of heavy metals, pesticides residues, and microorganisms in the ground AL rhizomes and the formulated(capsule) powder of AL, physicochemical and pharmaceutical properties of the formulated AL extract/capsule, and cytotoxicity evaluation of the formulated AL extract. Results of all evaluations confirmed satisfactory pharmaceutical properties of oral(capsule) formulation of the standardized AL extract.