Anti-inflammatory and wound healing effects of mouth gel containing kaempulchraol K from Kaempferia galanga rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia galanga L. is one of the important medicinal plants and has been used in Thailand for treating inflammation and wound. AIM OF THE STUDY: This study aimed to investigate the efficacy of the compound from K. galanga on wound healing and anti-inflammatory activities and develop a new product in gel form to maximize the benefits of this plant. MATERIALS AND METHOD: The mouth gel containing kaempulchraol K (KG2) was prepared by using 1.5% carbopol 934 as a gelling agent. Formulations of mouth gel containing KG2 at 0.10%, 0.25%, and 0.50% w/w were evaluated for color, smell, pH values, viscosity, and separation. Also, the chemical and biological stabilities of mouth gel containing KG2 were evaluated by heating-cooling test. The anti-inflammatory activity was tested against RAW 264.7 cells nitric oxide (NO) production and wound healing assay was performed using human gingival fibroblasts (HGF). RESULTS: Compound KG2 exhibited anti-NO production with an IC50 value of 66.8 µM and the wound healing activity of compound KG2 showed cell viability in the range of 90.9-111.4%. In addition, compound KG2 at a concentration of 3 µM induced the highest proportion of cell migration on day 3 at 90.2 ± 2.4%. The mouth gel containing KG2 both before and after the heating-cooling test exhibited good consistency, with pH values in the range of 6.64-6.71 (before) and 6.63-6.68 (after). Meanwhile, the viscosity was 81,700-96,700 cP (before) and 78,300-93,300 cP (after). For the chemical stability test of the active ingredient of mouth gel, the compound showed good stability after mixing with the gel base. The mouth gel exhibited anti-inflammation with IC50 values > 1000 µg/ml both before and after accelerating conditions. The wound healing activity of mouth gel containing KG2 (0.50% w/w) showed the highest % cell viability at 128.6% (before) and 123.8% (after). For cell migration, the result suggested that the mouth gel containing KG2 at 0.10%, 0.25%, and 0.50% w/w (3 µg/ml) on day 3 enhanced cell migration higher than that of the positive controls both before (85.0-96.8%) and after (and 84.4-94.3%) the accelerating conditions. CONCLUSION: The present study shows that mouth gel containing 0.50% KG2 is the most appropriate with good physical, chemical, and biological stabilities and might be one of the alternative sources for treatment of mouth ulcers (oral stomatitis) derived from aphthous ulcers, chemotherapy, and radiotherapy treatments.

Microbicide Containing Ellagic Acid Can Inhibit HIV-1 Infection.

OBJECTIVES: Ellagic acid (EA) has a wide range of biological effects. The purpose of this study was to investigate the in vitro effects of EA on HIV-1 replication, viral enzyme activity and cytokine secretion by infected cells. METHODS: The anti-HIV-1 activity of EA in solution was determined in vitro using the infection of TZM-bl cells by the nano luciferase-secreting R5-tropic JRCSF strain of HIV-1, which allows for the quantification of viral growth by measuring nano luciferase in the culture supernatants. The effect of EA on the cytokine secretion of TZM-bl cells was determined by a multiplexed bead array after 48 h of HIV-1 exposure. The antiviral effect of EA in the gel formulation (Ellagel), as would be used for vaginal application, was investigated by the inhibition of infection of UC87.CD4.CCR5 cells with R5-tropic pBaLEnv-recombinant HIV-1. RESULTS: EA in solutions of up to 100 µM was not toxic to TZM-bl cells. EA added either 1 h before or 4 h after HIV-1 exposure suppressed the replication of R5-tropic HIV-1 in TZM-bl cells in a dose-dependent manner, with up to 69% inhibition at 50 µM. EA-containing solutions also exhibited a dose-dependent inhibitory effect on HIV-1 replication in U87 cells. When EA was formulated as a gel, Ellagel containing 25 µM and 50 µM EA inhibited HIV-1 replication in U87 cells by 56% and 84%, respectively. In assays of specific HIV-1 enzyme activity, Ellagel inhibited HIV-1 integrase but not protease. EA did not significantly modulate cytokine secretion. CONCLUSIONS: We conclude that EA either in solution or in a gel form inhibits HIV infection without adverse effects on target cells. Thus, gel containing EA can be tested as a new microbicide against HIV infection.

Wound healing properties of pharmaceutical gel containing isopimarane diterpene isolated from Kaempferia galanga L.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia galanga L. rhizomes have been widely used in Thailand as medicine for treating inflammation and wound. A number of bioactive compounds have been isolated from the rhizomes of K. galanga and these compounds exhibited various pharmacological activities. AIM OF THE STUDY: The objective of this study is to investigate the wound healing properties of gel containing 6ß-acetoxysandaracopimaradiene-1α, 9α-diol (KG6), a compound from K. galanga. MATERIALS AND METHODS: KG6 gel formulations were prepared using 1.0% carbopol 940 as gelling agent. Three KG6 gel formulations (0.10, 0.25, 0.50% w/w) were subjected to heating-cooling test to determine their physical, chemical and biological stabilities. The wound healing properties of KG6 gel formulations were performed using RAW264.7 cells for anti-inflammatory effect, while their impact on cell proliferation and migration, collagen content and H2O2-induced oxidative stress was examined using human dermal fibroblasts (HDF). RESULTS: The pH, viscosity and general appearance after the heating-cooling test of the three prepared gels were stable in the acceptable range of gel formulation for skin. Gel containing 0.25% KG6 showed better chemical stability than other formulations. The 0.25% KG6 gel significantly increased cell viability (102.8%) and produced the highest HDF cell migration (91.9%) which was greater than that of Aloe vera gel (96.2, 78.4%, respectively). This gel exhibited anti-inflammatory activity via suppressing nitric oxide release and improved the viability of HDF cells against H2O2-induced oxidative stress. The 0.25% KG6 gels also increased collagen content in HDF cells. CONCLUSION: The gel formulation consisting of 0.25% KG6 with 1.0% of carbopol 940 was found to be a promising pharmaceutical gel for wound treatments due to marked wound healing properties.

Anti-inflammatory, wound healing and antioxidant potential of compounds from Dioscorea bulbifera L. bulbils.

BACKGROUND: Dioscorea bulbifera L. (Dioscoreaceae) has been traditionally used in Thai folk medicine as a diuretic and anthelmintic, for longevity preparations, and for wound and inflammation treatment. This plant is also commonly used in traditional Indian and Chinese medicines in the treatment of sore throat, gastric cancer, rectal carcinoma and goiters. However, the wound healing effects of the active compounds in this plant have not been investigated. OBJECTIVE: This study aimed to identify compounds responsible for the wound healing activity of D. bulbifera and determine their potential anti-inflammatory and antioxidant activities. METHODS: Crude extracts of D. bulbifera bulbils, their derived fractions and eleven purified compounds were tested for anti-inflammatory activity against LPS-induced NO production in RAW264.7 macrophages. The wound healing effects were evaluated via cell proliferation and migration assays using human dermal fibroblasts (HDFs), and the antioxidant effects were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical (•OH) scavenging activity assays. RESULTS: 15,16-Epoxy-6α-O-acetyl-8ß-hydroxy-19-nor-clero-13(16),14-diene-17,12;18,2-diolide (2), (+)-catechin (5), quercetin (6) and myricetin (11) exhibited significantly potent wound healing effects and promoted marked cell proliferation, resulting in % viabilities of 107.4-137.6, 121.1-151.9, 98.0-131.9, 90.9-115.9, respectively. Among them, (+)-catechin produced the highest % cell migration, resulting in 100.0% wound closure sooner (at day 2) than the other compounds. In addition, 1 µg/ml (+)-catechin significantly increased fibroblast migration by 2.4-fold compared to that in the control after 24 h. Regarding anti-inflammatory properties, kaempferol (7) and quercetin (6) decreased (p < 0.005) NO production, with IC50 values of 46.6 and 56.2 µM, respectively. In addition, the crude extracts, solvent fractions and flavonoid compounds were also found to possess marked antioxidant activity in both DPPH and •OH radical scavenging assays. CONCLUSIONS: These findings provide more evidence to support the traditional use of D. bulbifera for the treatment of wounds and inflammation.

Anti-inflammatory effect of isopimarane diterpenoids from Kaempferia galanga.

Two new isopimarane diterpenes, 1α-hydroxy-14α-methoxyisopimara-8(9),15-diene (7) and 1α,14α-dihydroxyisopimara-8(9),15-diene (9) and eight known isopimarane diterpenes including (-)-sandaracopimaradiene (1), 6ß-acetoxysandaracopimaradiene-9α-ol (2), sandaracopimaradiene-7ß,9α-diol (3), sandaracopimaradiene-1α,9α-diol (4), 6ß-acetoxysandaracopimaradiene-9α-ol-1-one (5), 6ß-acetoxysandaracopimaradiene-1α,9α-diol (6), 6ß,14α-dihydroxyisopimara-8(9),15-diene (8), and 6ß,14ß-dihydroxyisopimara-8(9),15-diene (10) were isolated from hexane fraction of Kaempferia galanga ethanol extract. Compounds 5, 6, 8, and 9 exerted the good anti-inflammatory effect on lipopolysaccharide-stimulated nitric oxide production from RAW264.7 cells with IC50 of 11.2, 7.7, 14.3, and 12.1 µM, respectively. These four compounds inhibited nitric oxide synthase (iNOS) mRNA expression. Compounds 5 and 6 also suppressed cyclooxygenase 2 (COX-2) mRNA expression; in addition, compound 6 had mild inhibitory effect on TNF-α mRNA. Among these compounds, 5 dramatically inhibited iNOS and COX-2 mRNA expression. The influential structures were proposed to be oxygen substitute at C-1, C-6, and α-OH at C-14.

Evaluation of Anti-HIV-1 Integrase and Anti-Inflammatory Activities of Compounds from Betula alnoides Buch-Ham.

Betula alnoides is a medicinal plant in Thai traditional longevity preparations. The crude extracts of this plant possess various biological activities. However, the isolated compounds from this plant have no reports of anti-HIV-1 integrase (IN) activity. Therefore, the present study aims to investigate the anti-HIV-1 integrase and anti-inflammatory effects of isolated compounds from this plant and predict the interaction of compounds with integrase active sites. From the bioassay-guided fractionation of the ethanol extract of B. alnoides stems using chromatographic techniques, five pentacyclic triterpenoid compounds were obtained. They are betulinic acid (1), betulin (2), lupeol (3), oleanolic acid (4), and ursolic acid (5). Compound 2 exhibited the most potent inhibitory activity against HIV-1 IN, with an IC50 value of 17.7 µM. Potential interactions of compounds with IN active sites were investigated using computational docking. The results indicated that active compounds interacted with Asp64, a residue participating in 3'-processing, and Thr66, His67, and Lys159, residues participating in strand-transfer reactions of the integration process. Regarding anti-inflammatory activity, all compounds exerted significant inhibitory effects on LPS-induced nitric oxide production (IC50 < 68.7 µM). Thus, this research provides additional scientific support for the use of B. alnoides in traditional medicine for the treatment of HIV patients.

Anti-inflammatory and wound healing effects of gel containing Kaempferia marginata extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia marginata Carey (Zingiberaceae family) has been traditionally used in the treatment of inflammation. The whole plant decoction is used for treatment of fever. Ethanol extracts which exert potent anti-inflammatory properties are selected for wound healing assay. AIM OF THE STUDY: This study aimed to investigate activities of the extract and gel formulation on anti-inflammatory and wound healing activities. The anti-inflammatory and wound healing properties from this plant could support its traditional uses and obtain a new pharmaceutical product with good physical, chemical and biological stabilities. MATERIALS AND METHODS: The anti-inflammatory activity was tested on anti-nitric oxide (NO) production using RAW264.7 cells and wound healing assay using human dermal fibroblast (HDF) cells. RESULTS: The results found that the anti-inflammatory activity of gel containing K. marginata at 10% w/w showed the highest activity with an IC50 value of 12.50 µg/ml (Diclofenac gel, IC50 value = 64.90 µg/ml). It also revealed that wound healing activities of K. marginata gel (5% w/w) showed the highest % cell viability (134.05%) and the highest % cell migration at 85.20 using HDF cells. CONCLUSION: The present study shows that gel containing K. marginata extracts have good anti-inflammatory and wound healing properties in vitro.

Anti-inflammatory and wound healing effects of cream containing Curcuma mangga extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma mangga Valeton & Van Zijp. (Zingiberaceae family) contains curcuminoids and diterpenes which have anti-inflammatory effect. In traditional use, the elixir of this plant has been used to detoxify the poisonous plants, treatment of gastric ulcer, chest pain, fever, skin disease and womb healing. This plant has also been reported for anti-inflammation in carrageenan-induced rat paw edema. AIM OF THE STUDY: This study is aimed to evaluate the anti-inflammatory and wound healing effects of cream containing C. mangga in the formulation as well as physical and chemical stabilities. MATERIALS AND METHODS: Three cream base formulas were evaluated for color, smell, pH values, viscosity and separation. The most stable cream base was chosen to mix with 2-10% of C. mangga extract. After that, the physical, chemical and biological properties of C. mangga cream before and after heating-cooling were evaluated. RESULTS: The results showed that C. mangga cream exhibited good consistency, a pH range of acid value (5.0-6.0) and the cream was stable. Cream containing 10% w/w C. mangga inhibited inflammation before and after accelerating conditions with IC50 values of 34.1 and 37.9 µg/ml which were better than 5% (IC50 = 42.9, 44.7 µg/ml) and 2% (IC50 = 49.1, 49.6 µg/ml), respectively. In addition, the anti-inflammatory effect of cream containing C. mangga was found to be better than diclofenac gel (IC50 = 54.3 µg/ml). Cream containing C. mangga before and after heating-cooling test at 1 and 3 µg/ml enhanced HDF viability of over 100%, while cream containing 5% w/w C. mangga before and after heating-cooling test enhanced migration of HDF cells at 36 h up to 75-80%. The curcuminoids which are curcumin, demethoxycurcumin and bisdemethoxycurcumin showed good chemical stability after heating-cooling test by using HPLC with the relative peak area % at 10:75:15, respectively. CONCLUSION: This study concluded that the development of cream containing C. mangga could reduce inflammation and heal the wound.

Antiinflammation constituents from Curcuma zedoaroides.

Curcuma zedoaroides, a Zingiberaceae species, has been used for snake bite antidote and wound care in Thailand. Seven compounds were isolated from the bioactive chloroform extract consisted of one new guaiane sesquiterpene lactone, 5-epiphaeocaulisin A (4) and one new diarylheptanoid, 1,2,3,5-tetrahydroxy-1-(4-hydroxy-3-methoxyphenyl)-7-(4-hydroxyphenyl) heptane (7), together with five known guaiane-type sesquiterpene lactones including gweicurculactone (1), zedoalactone B (2), phaeocaulisin C (3), zedoalactone H (5), and zedoalactone E (6). The antiinflammation was investigated on NO and TNF-α production using RAW264.7 cells. In addition, the expressions of genes involved in inflammation including iNOS, COX-2, and TNF-α were assessed. The results found that Compounds 1-7 presented their antiinflammation against NO production. The most potential effects were demonstrated on Compounds 1 and 7 with IC50 of 27.3 and 32.6 µM, respectively. Although, Compounds 1 and 7 did not inhibit TNF-α production, their suppression on iNOS and COX-2 mRNA expression were revealed. These results support the ability of chloroform fraction, Compounds 1 and 7 on antiinflammation, whereas others exhibited moderate and mild effect.

Anti-inflammatory 12,20-Epoxypregnane and 11,12-seco-Pregnane Glycosides from the Stems of Hoya kerrii.

Five 12,20-epoxypregnane glycosides (1-3, 5, and 6) and two 11,12-seco-pregnane glycosides (4 and 7) with spirodilactone motifs, as well as spirodilactone cleavage products 8 and 9, were isolated from the stems of Hoya kerrii. The relative configurations of the three related skeletons were supported by ROESY experiments and X-ray crystallographic analyses. The isolates were evaluated for their anti-inflammatory activity based on the inhibition of NO production in RAW264.7 cells, and some showed IC50 values ranging from 12.6 to 96.5 µM. The most potent compound, 9a, was also examined for its anti-inflammatory mechanism against mRNA expression and was found to down-regulate mRNA expression of iNOS and COX-2 in a dose-dependent manner.

Wound healing property of isolated compounds from Boesenbergia kingii rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Boesenbergia kingii have been traditionally used in the treatment of inflammatory bowel disease, ulcerative colitis, aphthous ulcer, stomach discomfort, dysentery and abscess. Previously, we reported the B. kingii extract exert potential wound healing properties. Therefore the search of responsible constituents for wound healing property from these rhizomes is still relevant. AIM OF STUDY: This study was aimed to investigate for wound healing property of compounds from this plant in order to support its traditional uses. MATERIAL AND METHODS: Wound healing activities were tested using in vitro assays including cell proliferation and migration assays, collagen production and H2O2-induced oxidative stress in mouse fibroblast L929 cells. The DPPH assay was also used to determine antioxidant activity. RESULTS: Fourteen compounds from the chloroform fraction possessed potent anti-oxidant and wound healing activities. Compound 11 exhibited the most potent anti-DPPH effect (IC50=21.0µM) and also active against 0.5mMH2O2-induced oxidative stress by increasing cell survival ability up to 60.3% at 10µM. In addition, compounds 3, 8 and 14 at 10µM significantly enhanced L929 viability with 119.2%, 122.7% and 113.7%, respectively. Compounds 2, 7, 8 and 14 markedly enhanced L929 migration on day 2 up to 60-76% at 10µM, whereas 7 and 14 strongly stimulated collagen production at 75.0 and 96.7µg/ml compared to the control group (57.5µg/ml), respectively. CONCLUSION: B. kingii is responsible for wound healing property via antioxidative effect, stimulation of fibroblast proliferation and migration as well as enhancement of collagen production.

Anti-HIV-1 integrase compounds from Dioscorea bulbifera and molecular docking study.

CONTEXT: Dioscorea bulbifera L. (Dioscoreaceae) has been used in a traditional Thai longevity medicine preparation. Isolation of inhibitors from natural products is a potential source for continuous development of new HIV-1 integrase (IN) inhibitors. OBJECTIVE: The objective of this study is to isolate the compounds and evaluate their anti-HIV-1 IN activity, as well as to predict the potential interactions of the compounds with an IN. MATERIALS AND METHODS: The ethyl acetate and water fractions (1-100 µg/mL) of Dioscorea bulbifera bulbils were isolated and tested for their anti-HIV-1 IN activity using the multiplate integration assay (MIA). The interactions of the active compounds with IN were investigated using a molecular docking method. RESULTS AND DISCUSSIONS: The ethyl acetate and water fractions of Dioscorea bulbifera bulbils afforded seven compounds. Among these, allantoin (1), 2,4,3',5'-tetrahydroxybibenzyl (2), and 5,7,4'-trihydroxy-2-styrylchromone (5) were isolated for the first time from this plant. Myricetin (4) exhibited the most potent activity with an IC50 value of 3.15 µM, followed by 2,4,6,7-tetrahydroxy-9,10-dihydrophenanthrene (3, IC50 value= 14.20 µM), quercetin-3-O-ß-D-glucopyranoside (6, IC50 value = 19.39 µM) and quercetin-3-O-ß-D-galactopyranoside (7, IC50 value = 21.80 µM). Potential interactions of the active compounds (3, 4, 6, and 7) with the IN active site were additionally investigated. Compound 4 showed the best binding affinity to IN and formed strong interactions with various amino acid residues. These compounds interacted with Asp64, Thr66, His67, Glu92, Asp116, Gln148, Glu152, Asn155, and Lys159, which are involved in both the 3'-processing and strand transfer reactions of IN. In particular, galloyl, catechol, and sugar moieties were successful inhibitors for HIV-1 IN.

Anti-HIV-1 integrase effect of compounds from Aglaia andamanica leaves and molecular docking study with acute toxicity test in mice.

CONTEXT: Acquired immunodeficiency syndrome (AIDS) is a serious health problem worldwide. It has been reported that Aglaia andamanica Hiern (Meliaceae) leaves possessed an antiviral effect. Therefore, a search of anti-HIV-1 integrase (HIV-1 IN) agents from A. andamanica is a promising target. OBJECTIVE: The objective of this study is to evaluate anti-HIV-1 IN activity of isolated compounds from A. andamanica using an in vitro assay and molecular docking study as well as testing acute toxicity in mice using the up and down method. MATERIALS AND METHODS: The leaves and compounds (3-100 µg/mL) from A. andamanica were determined for the anti-HIV-1 IN effect using the multiplate integration assay (MIA) by detection the absorbance of the final product, p-nitrophenol, at 405 nm. The molecular docking with the HIV-1 IN of the active compound N-methyl-trans-4-hydroxy-l-proline (10) was also studied. The Swiss albino mice were used for an acute toxicity test. RESULTS AND DISCUSSION: Among the isolated compounds, 10 showed marked anti-HIV-1 IN effect with an IC50 value of 11.8 µg/mL, whereas other compounds were inactive (IC50 value > 100 µg/mL). The molecular docking of compound 10 with an HIV-1 IN enzyme was also studied. The result revealed that this compound formed the hydrogen bonding with the Thr66, Asn155, and Lys159 of the HIV-1 IN binding site. The acute toxicity of the A. andamanica extract was not observed at the dose 2000 mg/kg mice. This is the first report of A. andamanica for anti-HIV-1 IN activity.

Anti-HIV-1 integrase activity and molecular docking of compounds from Albizia procera bark.

CONTEXT: Albizia procera (Roxb.) Benth. (Mimosaceae) has been traditionally used in Thai longevity preparations. Thus, searching for HIV-1 integrase (HIV-1 IN) agents from natural sources is of interest. OBJECTIVE: The objective of this study is to examine the inhibitory activity against HIV-1 IN of compounds isolated from the stem bark of Albizia procera. MATERIALS AND METHODS: The EtOH extract and isolated compounds of Albizia procera bark were examined for anti-HIV-1 IN activity at various concentrations (10-100 µg/mL and 10-100 µM) using the multiplate integration assay and molecular docking. RESULTS AND DISCUSSIONS: The results showed that the ethanol extract had good anti-HIV-1 IN activity with an IC50 value of 19.5 µg/mL, whereas ethyl acetate fraction exhibited the most potent with an IC50 value of 19.1 µg/mL, followed by water fraction (IC50 value = 21.3 µg/mL), hexane and chloroform fractions (IC50 value > 100 µg/mL), respectively. From bioassay-guided isolation, the ethyl acetate fraction was further separated to give two compounds which are (+)-catechin (1) and protocatechuic acid (2), respectively. Of the tested samples, (+)-catechin (1) exhibited appreciable activity against HIV-1 IN with an IC50 value of 46.3 µM, whereas protocatechuic acid (2) showed mild activity with 46.0% inhibition at concentration of 100 µM. (+)-Catechin (1) could interact with Thr66, Gly148, and Glu152 in the core domain of IN enzyme, whereas protocatechuic acid (2) could bind with Thr66, His67, Glu152, Asn155, and Lys159. This is the first report on anti-HIV-1 IN activity of Albizia procera bark. These results may suggest that Albizia procera bark has potential as anti-HIV-1 IN agent.

Antiinflammatory and Wound Healing Effects of Caesalpinia sappan L.

Extracted compounds from Caesalpinia sappan L. were examined for the inhibitory activity against NO, PGE2 , and TNF-α productions and on associated transcription levels using RAW264.7 cells. They were also tested for their effects on wound healing using fibroblast L929 cells. Among the compounds tested, brazilin (8) was the most effective against lipopolysaccharide (LPS)-induced NO production in RAW264.7 cells with an IC50 value of 10.3 µM, followed by sappanchalcone (2, 31.0 µM). Brazilin (8) also inhibited PGE2 and TNF-α production with IC50 values of 12.6 and 87.2 µM, respectively. The antiinflammatory mechanism of brazilin involved down regulation of the mRNA expressions of the iNOS, COX-2, and TNF-α genes in a dose-dependent manner. An ethanol (EtOH) extract of C. sappan significantly increased fibroblast proliferation, fibroblast migration, and collagen production, whereas brazilin (8) only stimulated fibroblast migration. In addition, the EtOH extract showed no acute toxicity in mice, and it was therefore safe to make use of its potent antiinflammatory and wound healing activities. Brazilin was mainly responsible for its antiinflammatory effect through its ability to inhibit the production of NO, PGE2 , and TNF-α. This study supports the traditional use of C. sappan for treatment of inflammatory-related diseases.

Anti-HIV-1 Integrase Activity and Molecular Docking Study of Compounds from Caesalpinia sappan L.

Caesalpinia sappan L. (Caesalpiniaceae) has been traditionally used as blood tonic, expectorant, and astringent by boiling with water. Searching for HIV-1 integrase (IN) inhibitors from this plant is a promising approach. The EtOH extract of C. sappan and its isolated compounds were tested for their anti-HIV-1 IN effect using the multiplate integration assay, and the active compounds were determined for their mechanisms by molecular docking technique. Extraction from the heartwoods and roots of C. sappan led to the isolation of nine compounds. Among the compounds tested, sappanchalcone (2) displayed the strongest effect against HIV-1 IN with an IC50 value of 2.3 µM followed by protosappanin A (9, IC50 = 12.6 µM). Structure-activity relationships of compounds from C. sappan were found, in which the vicinal hydroxyl moiety were essential for anti-HIV-1 IN effect of compounds 2 and 9 by binding with the amino acid residues Gln148 and Thr66 in the core domain of the HIV- 1 IN enzyme, respectively.

Use of a hexasubstituted benzene scaffold in the development of multivalent HIV-1 integrase inhibitors.

The highly directional hexasubstituted benzene moiety was used as the central scaffold to create new human immunodeficiency virus (HIV)-1 integrase inhibitors through the attachment of multiple active groups. A series of potential inhibitors having substituted polyhydroxylated mono, bis and tris-cinnamoyl derivatives connected on the scaffold were prepared through Claisen-Schmidt condensations with substituted benzaldehydes, followed by partial demethylation to uncover the active phenolic groups required for the interactions with the integrase enzyme active sites. Using a multiplate integration assay method, four compounds carrying at least two sets of interacting moieties were found to be relatively potent integrase inhibitors with IC50 values in the low micromolar range. The results confirmed that multiple polyhydroxylated groups were required on the platform in order to effectively interact with the enzyme. The results from molecular docking studies consistently complemented the experimental results and revealed the nature of the potential key binding interactions responsible for the apparent activity of the active compounds.

Anti-inflammatory activity of compounds from Boesenbergia longiflora rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: The rhizomes of Boesenbergia longiflora (Wall.) Kuntze have been traditionally used in treatment of inflammatory bowel disease, ulcerative colitis, aphthous ulcer and abscess. Our previous study indicated that CHCl3 fractions of Boesenbergia longiflora had potential on anti-inflammatory properties. In the present study, we investigated the active constituents of this plant for anti-inflammatory activity in order to support its traditional use. MATERIAL AND METHODS: The CHCl3 fraction was isolated using chromatographic techniques. Isolated compounds were tested using relevant in vitro anti-inflammatory assays against LPS-induced NO and TNF-α releases as well as their mechanisms in transcription levels in murine macrophage RAW264.7 cells. RESULTS: The isolation of the CHCl3 fraction from Boesenbergia longiflora rhizomes led to the isolation of three new daucane sesquiterpenes, which were identified as 8-hydroxy-dauca-9, 11-diene-7-one (longiferone A; 1), dauca-8, 11-diene-7-one (longiferone B; 2) and dauca-8, 11-diene-7, 10-dione (longiferone C; 3); together with four known flavonoids, six known diarylheptanoids as well as one sterol. The longiferone B (2) and longiferone C (3) showed anti-inflammatory activity against NO release with IC50 values of 21.0 and 31.3µM, respectively. Longiferone B (2) also suppressed the iNOS and COX-2 mRNA expression. Moreover, the flavonoids and diarylheptanoids inhibited NO and TNF-α production in a dose dependent manner. CONCLUSION: This study demonstrated that sesquiterpenes, diarylheptanoids and some methoxyflavonoids found in Boesenbergia longiflora are responsible for anti-inflammatory activity.

Inhibition of nitric oxide production in lipopolysaccharide-activated RAW264.7 macrophages by isolated xanthones from the roots of Cratoxylum formosum ssp. pruniflorum.

The inhibitory activity of extract and compounds isolated from the roots of Cratoxylum formosum ssp. pruniflorum against nitric oxide (NO) was evaluated using RAW264.7 cells. Isolation of the CH2Cl2 extract of C. formosum ssp. pruniflorum roots afforded ten known xanthones including six tri-oxygenated xanthones (1-6) and four tetra-oxygenated xanthones (7-10), respectively. Compound 7 showed the highest inhibitory activity against NO release with an IC50 value of 3.9 µM, followed by compound 8 with an IC50 value of 4.3 µM, respectively. In order to understand the mechanism of this anti-inflammatory activity, the transcriptional level of 7 was found to down regulate mRNA expressions of iNOS and COX-2 in dose-dependent manners, whereas 8 inhibited only iNOS mRNA expression but did not affect on that of COX-2 gene. Xanthones might be the main anti-inflammatory components in C. formosum ssp. pruniflorum.

Nitric oxide and tumor necrosis factor-alpha inhibitory substances from the rhizomes of Kaempferia marginata.

The ethanol extract of the rhizomes of Kaempferia marginata showed a potent inhibitory effect against lipopolysaccharide (LPS)-induced nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) release in RAW264.7 cells. Moreover, the partition with various organic solvents also inhibited NO production. One new pimarane-type diterpene, 1alpha-acetoxysandaracopimaradien-2alpha-ol (5), along with four known diterpenes (1-4), were isolated from the n-hexane and chloroform layers, respectively. Among these metabolites, compounds 1 and 4 were isolated for the first time from K. marginata. Compounds 1-5 showed significant inhibitory effects on NO production, with IC50 values ranging from 38.6 to 51.9 microM. Furthermore, compound 2 also exhibited significant activity against TNF-alpha release (IC50 = 48.3 microM). These findings may support the use of K. marginata by traditional doctors for treatment of inflammatory-related diseases.