HM Medical Clinic


Microsoft word - re-revised paper_planta medica-20110721-final.doc

This is an author produced version of a paper published in Planta Medica
This paper has been peer-reviewed but does not include the final publisher proof- corrections or journal pagination. Citation for the published paper: Endale, Milkyas; Alao, John Patrick; Akala, H. M.; Rono, N. K.; Eyase, F. L.; Derese, S.; Ndakala, A.; Mbugua, M.; Walsh, D. S.; Sunnerhagen, Per; Erdelyi, Mate; Yenesew, Abiy Antiplasmodial Quinones from Pentas longiflora and Pentas lanceolata
Planta Medica, 78 ( 1 ) s. 31-35 Access to the published version may require subscription. Published with
permission from: Thieme
Gothenburg University Publications Research article:
Antiplasmodial Quinones from Pentas longiflora
and Pentas lanceolata
Planta Medica 2012, 78, 31-35
The publisher's copyright information is available on:
The publisher's version of the article is available on:
Antiplasmodial Quinones from Pentas longiflora and Pentas lanceolata Milkyas Endale, 1 John Patrick Alao, 2 Hoseah M. Akala, 3 Nelson K. Rono, 1 Fredrick L.
Eyase,3 Solomon Derese,1 Albert Ndakala, 1 Martin Mbugua, 1 Douglas S. Walsh, 3 Per
Sunnerhagen, 2 Mate Erdelyi, 4* Abiy Yenesew1*
1Department of Chemistry, University of Nairobi, P.O. Box 30197-00100, Nairobi, Kenya.
2Department of Cell- and Molecular Biology, University of Gothenburg, SE-405 30 Gothenburg, Sweden. 3United States Army Medical Research Unit-Kenya, MRU 64109, APO, AE 09831- 4109, USA. 4Department of Chemistry, University of Gothenburg, SE-412 96 Gothenburg, Sweden and the Swedish NMR Centre, University of Gothenburg, P.O. Box 465, SE-40530 Gothenburg, Sweden CORRESPONDENCE
* (AbiyYenesew) Tel/Fax: +254-02-4446138, E-mail: [email protected]. (Mate Erdelyi)
Tel: +46-31-7869033, E-mail: [email protected] ABSTRACT The dichloromethane/methanol (1:1) extracts of the roots of Pentas longiflora and
Pentas lanceolata showed low micromolar (IC50 = 0.9-3µg/mL) in vitro antiplasmodial activity against chloroquine-resistant (W2) and chloroquine-sensitive (D6) strains of Plasmodium falciparum. Chromatographic separation of the extract of Pentas longiflora led to the isolation of the pyranonaphthoquinones pentalongin (1) and psychorubrin (2) with IC50 values below 1
µg/mL and the naphthalene derivative mollugin (3) which showed marginal activity. Similar
treatment of Pentas lanceolata led to the isolation of eight anthraquinones (4-11, IC50 = 5-
31µg/mL) of which one is new (5,6-dihydroxydamnacanthol, 11) while three - nordamnacanthal
(7), lucidin-ω-methyl ether (9), and damnacanthol (10) - are reported here for the first time from
the genus Pentas. The compounds were identified by NMR and mass spectroscopic techniques. KEYWORDS Pentas longiflora, Pentas lanceolata, Rubiaceae, anthraquinone, 5,6-
dihydroxydamnacanthol, pyranonaphthoquinone, malaria According to the estimates of the World Health Organization almost one million deaths are caused by malaria each year in Africa alone, of which most are children under the age of five [1]. In addition, this mosquito-borne disease has a serious economic impact due to loss of commercial and labor outputs predominantly in countries with tropical and subtropical climates. Over 300, 000, 000 people worldwide are infected, and each year, nearly one third of these exhibit acute manifestations of the disease [2]. While awaiting the development of a malaria vaccine, millions of lives are still dependent upon treatment with chemotherapeutic agents. Since most of the available drugs are becoming increasingly ineffective due to the rapid emergence of resistant Plasmodium falciparum strains [3], there is an urgent need for novel antimalarial agents. Because of the high cost of the few still effective antimalarial drugs [4] traditional medicine remains an important source of treatment in developing countries. Pentas longiflora Oliver (Rubiaceae) is an important medicinal plant of Tropical East Africa [5]. In Kenya, a decoction of its roots mixed with milk is taken as a cure for malaria [6]. Although, its leaves have previously been tested for in vitro antimalarial activity; no attempts were made to isolate and identify the antiplasmodial constituents [7]. Pentas lanceolata (Forsk.) is mostly found in the highlands of Kenya and was reported to exhibit micromolar in vitro antiplasmodial activity against P. falciparum [8]. Although extracts of these plants have been assayed against a range of diseases [8, 9], their constituents have not been investigated for antiplasmodial activity. Motivated by the traditional uses and the preliminary screening reports [7, 8, 9], we performed isolation, characterization and antiplasmodial investigation of naphthoquinones and anthraquinones found in the extracts of the roots of P. longiflora and P. lanceolata. MATERIALS AND METHODS
General experimental procedures: Column chromatography was performed on oxalic acid
impregnated silica gel [the silica gel was deactivated by mixing 2 kg of silica gel 60 (70-230 mesh) with 3% oxalic acid (30 g in 1L water) and allowed to stand for 30 min, filtered and dried in an oven (100oC) for 45 min]. TLC was done using silica gel 60 F254 (Merck) precoated plates NMR analyses were carried out on Varian 800, 600, 500 and 200 MHz spectrometers. Structural assignment was performed based on gCOSY, gTOCSY, gNOESY, gHSQC, gHMBC, and gH2BC spectra. ESI LC-MS was performed on a Perkin Elmer PE SCIEX API 150 EX instrument equipped with a Turbolon spray ion source and a Gemini 5 mm C18 110 Å HPLC column using a water-acetonitrile gradient (80:20 to 20:80). High-resolution mass spectral analysis (Q-TOF-MS) was performed at Stenhagen Analyslab AB, Gothenburg, Sweden. Compound purity was determined by NMR and HPLC. Analytical HPLC was run on a Hewlett Packard Series 1050 HPLC using the Software Chromulan (Pikron Ltd), a Gemini 5 mm C18 110 Å HPLC column and methanol-water mixture as eluent. Plant material: The roots of Pentas longiflora were collected from Nandi East district, Kenya
(Nandi Hills-Chebarus location) in August, 2009. The roots of Pentas lanceolata were collected from Ngong forest in December, 2009. The plant materials were identified by Mr Patrick Chalo Mutiso, School of Biological Sciences, University of Nairobi. Specimens are deposited at the Herbarium, School of Biological Sciences, University of Nairobi under voucher numbers MEA 2009/001 (Pentas longiflora) and MEA 2009/002 (Pentas lanceolata). Extraction and isolation: The dried and grounded roots of Pentas longiflora (1.1 kg) were
extracted by cold percolation with CH2Cl2:MeOH (1:1) three times for 24 hrs in each case. The extract was concentrated using a rotary evaporator to yield a brownish crude extract (50 g, 4.54 %). A 35 g portion of the crude extract was subjected to column chromatography (80 cm length and 80 mm diameter column size, 350 g oxalic acid impregnated silica gel) with increasing gradient of acetone in n-hexane. Two hundred fractions (each ca. 200 mL) were collected. Fractions 15-17 (2% acetone in n-hexane) were purified by Sephadex LH-20 (eluent CH2Cl2:MeOH; 1:1) to give mollugin (3, 34 mg). Fractions 18-25 (3% acetone in n-hexane) were
purified by column chromatography on oxalic acid impregnated silica gel (eluent, 2% acetone in n-hexane) to give pentalongin (1, 40 mg). Fractions 90-112 (20% acetone in n-hexane) were
combined and purified by Sephadex LH-20 (eluent, CH2Cl2/MeOH; 1:1) to give psychorubrin (2,
The ground roots (1.4 kg) of Pentas lanceolata were extracted with CH2Cl2:MeOH (1:1) and then with methanol three times for 24 hrs in each case. The extracts were concentrated using a rotary evaporator to yield a brownish crude extract (57 g, 4.8%) and (100 g, 7.1%), respectively. A 54 g portion of the crude CH2Cl2:MeOH (1:1) extract was subjected to column chromatography (80 cm length and 80 mm diameter, 420 g oxalic acid impregnated silica gel) with increasing gradient of ethyl acetate in n-hexane. A total of 550 fractions (each 200 mL) were collected. Fractions 10-13 (2% ethyl acetate in n-hexane) were combined and purified on Sephadex LH-20 (eluent, CH2Cl2:MeOH; 1:1) to give tectoquinone (4, 40 mg). Fractions 15-25
(eluent, 3% ethyl acetate in n-hexane) were combined and purified by Sephadex LH-20 (eluent, CH2Cl2:MeOH; 1:1) to give rubiadin (5, 680 mg) and rubiadin-1-methyl ether (6, 50 mg).
Fractions 30-35 (5% ethyl acetate in n-hexane as eluent) were combined and purified by column chromatography to give damnacanthal (8, 320 mg). Fractions 53-65 (7% ethyl acetate in n-
hexane) were combined and purified on Sephadex LH-20 with CH2Cl2:MeOH; (1:1) as eluent to give nordamnacanthal (7, 20 mg). Fractions 131-135 (18% ethyl acetate in n-hexane) were
combined and purified using column chromatography on oxalic acid impregnated silica gel (increasing gradient of ethyl acetate in n-hexane) to give lucidin-ω-methyl ether (9, 50 mg).
Fractions 400-405 (50% ethyl acetate in n-hexane) were combined and purified by MPLC (increasing gradient of ethyl acetate in n-hexane as eluent, flow rate of 30 mL/min) to give damnacanthol (10, 50 mg).
The methanol extract (70 g) was subjected to column chromatography on oxalic acid impregnated silica gel (80 cm length and 80 mm diameter, 500 g oxalic acid impregnated silica gel) eluting with increasing gradient of methanol in dichloromethane. A total of 100 fractions (each ca. 200 mL) were collected. Fractions 5-11 (100% dichloromethane) were combined and purified on Sephadex LH-20 (eluent, CH2Cl2:MeOH; 1:1) to give rubiadin (5, 20 mg) and
rubiadin-1-methyl ether (6, 18 mg). Fractions 21-25 (eluent, 1% of methanol in CH2Cl2) were
combined and further purified on Sephadex LH-20 (eluent, CH2Cl2:MeOH; 1:1) to give damnacanthol (10, 15 mg). Fractions 87-90 (5% MeOH in CH2Cl2) were combined and purified
using Sephadex LH-20 (eluent, CH2Cl2/MeOH; 1:1) to give 5,6-dihydroxydamnacanthol (11, 40
Drugs: The reference antimalarial drugs, chloroquine and mefloquine having well-documented
IC50 values were tested alongside test samples pyranonaphthoquinones and a naphthalene derivative isolated from the roots of Pentas longiflora; anthraquinones isolated from the roots of Pentas lanceolata as described above. Drug susceptibility testing: Two laboratory clones of Plasmodium falciparum, the Sierra Leone
D6 chloroquine-sensitive and the Indochina W2 chloroquine-resistant were maintained in continuous culture to attain replication robustness prior to assays. Drug susceptibility was tested by the Malaria SYBR Green I-based in vitro assay technique described in Juma et al. [10]. Cytotoxicity assay: Experimental details are given in the supporting information.
5,6-Dihydroxydamnacanthol (11). Red solid. UV (CH3OH) max 218, 274, 308, 424, nm. 1H
NMR (Table 2). 13C NMR (Table 2) HRMS (ESI): m/z = 317.0659 [M+H]+, calcd. 317.06558. RESULTS AND DISCUSSION
In our hands, the root extracts of P. longiflora and P. lanceolata showed significant antiplasmodial activities (Table 1). From the root extract of P. longiflora the naphthoquinone derivatives pentalongin (1) [11, 12, 13], psychorubrin (2) [14] and mollugin (3) [15, 13] were
chromatographically isolated, identified and tested for antiplasmodial activities. The major constituents 1 and 2 showed good to moderate activities (IC50 < 1 µg/mL) whereas 3 has
marginal inhibition against the W2 chloroquine-resistant and D6 chloroquine-sensitive strains of P. falciparum (Table 1). Although these compounds were previously reported [11, 12] and studied for antibacterial [16], antifungal [17], and antiviral [18] properties, their antiplasmodial activities are reported here for the first time. Chromatographic separation of the dichloromethane/methanol (1:1) extract of the roots of P. lanceolata resulted in the isolation of seven known anthraquinones (Figure 2), spectroscopically (NMR and MS) identified as tectoquinone (4) [15], rubiadin (5) [19], rubiadin-1-methyl ether (6)
[19], nordamnacanthal (7) [20], damnacanthal (8) [19], lucidin-ω-methyl ether (9) [26, 29], and
damnacanthol (10) [21]. Three of these (7, 9 and 10) are reported here for the first time from the
genus Pentas. In agreement with previous investigations on rubiadin-1-methyl ether (6),
damnacanthal (8), and lucidin-ω-methyl ether (9) [22], the anthraquinones isolated from the roots
of P. lanceolata showed moderate antiplasmodial activities (Table 1). The methanol extract yielded further amounts of 4, 5, 10 and a new compound 11 (Figure 3)
isolated as a red solid. The Q-TOF-MS spectrum provided the exact mass at m/z 317.0659 [M+H]+, suggesting a molecular formula of C16H12O7. The UV-VIS absorption maxima at 218, 274, 308 and 424 nm suggests a 9,10-anthraquinone skeleton [23]. Its 1H NMR spectrum (Table 2) revealed an aromatic singlet, a pair of ortho-coupled aromatic protons, a methoxy and an oxymethylene substituent as well as three solvent accessible and one chelated (H 12.40) hydroxyl groups. Two carbonyl functionalities were indicated by 13C-NMR. HMBC correlation of the methoxy protons with C-1, the oxymethylene protons with C-1, C-2 and C-3 (Table 2) are consistent with the methoxy, oxymethylene, and a hydroxyl substitution in ring A. The high chemical shift of the methoxyl group (C 62.4 ppm) is indicative of di-ortho [24] substitution allowing its placement at C-1 rather than C-3. Hence, in similarity to previously identified anthraquinones of the Rubiaceae family [25], ring A of 11 is oxygenated at C-1 and C-3 and has
the oxymethylene at C-2. The aromatic singlet at H 7.55 ppm (H-4) showed HMBC correlation with the C-10 carbonyl (C 189.2 ppm), indicating their peri position. The high chemical shift of this carbonyl is indicative of a peri hydroxyl group at C-5, which is further confirmed by the HMBC correlation of the aromatic doublet at H 7.57 ppm (H-8) to the carbonyl at C 178.7 ppm (C-9), but not with the one at C 188.5 ppm (C-10). These three bond heteronuclear correlations confirm the dihydroxy-substitution at C-5 and C-6 in ring C. Therefore, compound 11 was
characterized as 3,5,6-trihydroxy-1-methoxy-2-hydroxymethyl-9,10-anthraquinone (Figure 3) for which the trivial name 5,6-dihydroxydamnacanthol is proposed. Our assignation is in good agreement with that of the recently reported and closely-related 2-hydroxymethyl-1-methoxy- 3,5,6-trihydroxyanthraquinone-3-O--glycopyranoside, isolated from Putoria calabrica (L. fil, Rubiaceae) [26]. An additional evidence for the biosynthetic route in the family Rubiaceae [25] yielding compound 11 is the presence of the 2-ethoxyl-derivative of 11, 2-ethoxymethyl-3,5,6-
trihydroxy-1-methoxyanthraquinone, in the extract of Putoria calibrica (L. fil) [27]. Based on the biosynthesis of anthraquinones of the Rubiaceae, most of these compounds are substituted with hydroxyl, methoxyl and/or methyl groups in ring A (Figure 2) [25], and some carry additional hydroxyl or alkoxyl groups in ring C, mainly at positions 5 and 6 [25, 28]. These latter oxygen atoms are introduced at a late stage of the biogenesis [25] which is shown for example for morindone, as reported from the cell cultures of Morinda citrifolia [29] and for putorinoside A, isolated from Putoria calábrica [27]. As a consequence of the biosynthetic pathway, most, if not all, anthraquinones carry a carbon substituent at position 2 in ring A [25]. One of the rare exceptions from the above rule is 2-ethoxy-1-hydroxyanthraquinone isolated from Morinda citrifolia [30], a compound lacking carbon (CH2, CHO, CH, etc) substitution at C- 2. We would like to emphasize that if carbon substitution is present in an anthraquinone derived from the family Rubiaceae, based on biogenetics [25] the currently accepted nomenclature it is placed unambiguously at position 2 in ring A. Not following the above convention [31] may be perplexing in the evaluation of biosynthetic routes and bioactivities. Hence, the compounds anthraquinone [31] should be correctly named as 5,6-dimethoxy-2-methyl-9,10-anthraquinone and 6-hydroxy-5-methoxy-2-methyl-9,10-anthraquinone. Since complete and correctly assigned spectroscopic characterization was not available for several anthraquinones described here, detailed MS, and 1H and 13C NMR analysis (based on homo- and heteronuclear correlation spectra providing unambiguous assignment) is reported (Supporting information). Despite their promising activity against the W2 and D6 strains of Plasmodium falciparum, the comparably high cytotoxicity (Table 1) of 1 and 2 makes their direct application as antimalarial
agents virtually impossible. The anthraquinones isolated from Pentas lanceolata, 4-11, show low
cytotoxicity indicating the safer applicability of the anthraquinone containing indigenous decoction of P. lanceolata as compared to that of the pyranonaphtoquinone containing P. longiflora. In conclusion, the pyranonaphthoquinones and some of the anthraquinones isolated from the roots of P. lanceolata and P. longiflora showed good to moderate antiplasmodial activities against the W2 and D6 strains of Plasmodium falciparum, and an overall low cytotoxicity for anthraquinones. Careful analysis of their structure-activity relationship followed by rational synthetic modifications has potential for identifying more applicable agents in the fight against ACKNOWLEDGEMENT M. Endale is thankful to the German Academic Exchange service
(DAAD) and the Natural Products Research Network for Eastern and Central Africa (NAPRECA) for a Ph.D. Scholarship. J.P. Alao and P. Sunnerhagen are thankful to the Chemical Biology Platform at the University of Gothenburg. M. Erdelyi is thankful for the financial support of the Swedish Research Council (VR2007-4407) and the Royal Society of Arts and Sciences in Göteborg. Herewith we declare the absence of any conflict of interest, financial or personal, for all authors.
1 WHO, World Malaria Report 2008, Geneva, 2008: 9.
2 Heyneman D. The Worldwide Burden of Parasitic Disease, in Parasitic Infections, J. Leech, M. Sande and R. Root, Eds. Churchill Livingstone: New York. 1988: 11-32. 3 Wongsrichanalai C, Pickard AL, Wernsdorfer WH, Meshnick SR. Epidemiology of drug- resistant malaria. Lancet Infect Dis 2002; 2: 209-218. 4 Willox ML, Bodeker, G. Traditional herbal medicines for malaria. Br Med J 2004; 329: 5 Njoroge NG, Bussmann WR, Gemmill B, Newton LE, Ngumi VW. Utilisation of weed species as sources of traditional medicines in central Kenya. Lyonia 2004; 7: 71-87. 6 Kokwaro JO. Medicinal Plants of East Africa; University of Nairobi press, Nairobi, 2010: 7 Wanyoike GN, Chhabra SC, Lang'at-Thoruwa CC, Omar SA. Brine shrimp toxicity and antiplasmodial activity of five Kenyan medicinal plants. J Ethnopharm 2004; 90: 129-133. 8 Koch A, Tamez P, Pezzuto J, Soejarto D. Evaluation of plants used for antimalarial treatment by the Massai of Kenya. J Ethnopharm 2005; 101: 95-99. 9 Nayak BS, Vinutha B, Geetha B, Sudha B. Experimental evaluation of Pentas lanceolata flowers for wound healing activity in rats. Fitoterapia 2005; 76: 671-675. 10 Juma WP, Akala HM, Eyase FL, Muiva LM, Heydenreich M, Okalebo FA, Gitu PM, Peter MG, Walsh DS, Imbuga M, Yenesew A. Terpurinflavone: An antiplasmodial flavone from the stem of Tephrosia Purpurea. Phytochem Lett 2011; 4: 176-178. 11 De Kimpe N, Van Puyvelde L, Schripsema J, Erkelens C, Verpoorte R. Complete Proton and Carbon-13NMR Spectral Assignments of Pentalongin. Magn Res Chem 1993; 31: 329- 12 Hari L, De Buyck LF, De Pooter HL. Naphthoquinoid Pigments from Pentas longiflora. Phytochemistry 1991; 30: 1726-1727. 13 El Hady S, Bukuru J, Kesteleyn B, van Puyvelde L, De Kimpe N, Van TN. New pyranonaphthoquinone and pyranonaphthohydroquinone from the roots of Pentas longiflora. J Nat Prod 2002; 65: 1377-1379. 14 Hayashi T, Smith FT, Lee KH. Antitumor agents. 89. Psychorubrin, a new cytotoxic naphthoquinone from Psychotria rubra and its structure-activity relationships. J Med Chem 1987; 30: 2005-2008. 15 Liu R, Lu Y, Wu T, Pan Y. Simultaneous Isolation and Purification of Mollugin and Two Anthraquinones from Rubia cordifolia by HSCCC. Chromatographia 2008; 68: 95-99 16 Van T, De Kimpe N. Synthesis of pyranonaphthoquinone antibiotics involving the ring closing metathesis of a vinyl ether. Tetrahedron Lett 2004; 45: 3443-3446. 17 Puyvelde LV, Geysen D, Ayobangira FX, Hakizamungu E, Nshimyimana A, Kalise A. Screening of medicinal plants of Rwanda for acaricidal activity. J Ethnopharmacol 1985; 18 Ho LK, Don MJ, Chen HC, Yeh SF, Chen JM. Inhibition of hepatitis B surface antigen secretion on human hepatoma cells. Components from Rubia cordifolia. J Nat Prod 1996; 19 Kusamba C, Federici E, De Vicente Y, Galeffi C, The anthraquinones of Pentas zanzibarica. Fitoterapia 1993; 64:18-22. 20 Adesogan EK. Anthraquinones and anthraquinols from Morinda lucida: The biogenetic significance of oruwal and oruwalol. Tetrahedron 1973; 29: 4099-4102. 21 Ferrari F, Monache GD, Alves de Lima R. Two Naphthopyran Derivatives from Faramea cyanea. Phytochemistry 1985; 24: 2753-2755. 22 Koumaglo K, Gbeassor M, Nikabu O, De Souza C, Werner W. Effects of three compounds extracted from Morinda lucida on Plasmodium falciparum. Planta Med 1992; 58: 533-534 23 Scott AI. Interpretation of the Ultraviolet Spectra of Natural Products. Pergamon Press; Oxford: 1964; 286-289. 24 Schripsema J, Dagnino D. Elucidation of the substitution pattern of 9,10-anthraquinones through the chemical shifts of peri-hydroxyl protons. Phytochemistry 1996; 42: 177-184. 25 Han YS, Van der Heijen R, Verpoorte R. Biosynthesis of anthraquinones in cell cultures of Rubiaceae, Plant Cell, Tissue and Organ culture 2001; 67: 201-220. 26 Ealis I, Tasdemir D, Ireland CM, Sticher O. Two new Lucidin-type anthraquinone glycosides from Putoria calabrica. Chem Pharm Bull 2002; 50: 701-702. 27 Gonzalez A, Barrosso JT, Cardona RJ, Medina JM, Rodriguez Luis F. Química de la Rubiáceas. II. Componentes de la "Putoria calábrica" Perss. Anales de Quimica 1977; 73: 28 Schripsema J, Ramos VA, Verpoorte R. Robustaquinones, novel anthraquinones from an elicited Cinchona robusta suspension culture. Phytochemistry 1999; 51: 55-60. 29 Leistner E. Biosynthesis of morindone and alizarin in intact plants and cell suspension cultures of Morinda citrifolia. Phytochemistry 1973; 12: 1669-1674. 30 Ee GCL, Wen YP, Sukari MA, Go R, Lee HL. A new anthraquinone from Morinda citrifolia roots. Nat Prod Res 2009; 23: 1322-1329. 31 Osman CP, Ismail NH, Ahmad R, Ahmat N, Awang K, Jaafar FM. Anthraquinones with Antiplasmodial Activity from the Roots of Rennellia elliptica Korth. (Rubiaceae). Molecules 2010; 15: 7218-7226. Table 1. In vitro antiplasmodial activity and cytotoxicity of crude extracts and pure compounds
Sample (purity in %)
Antiplasmodial activity
50* (µg/mL)
Pentas longiflora (root extract) Pentalongin (1, ≥ 98%)
Psychorubrin (2, ≥ 98%))
Mollugin (3, ≥ 95%)
Pentas lanceolata (root extract) Tectoquinone (4, ≥ 98%)
Rubiadin (5, ≥ 98%)
Rubiadin-1-methyl ether (6, ≥ 98%)
Nordamnacanthal (7, ≥ 99%)
Damnacanthal (8, ≥ 99%)
Lucidin-ω-methyl ether (9, ≥ 98%)
Damnacanthol (10, ≥ 98%)
5,6-Dihydroxydamnacanthol ﴾11, > 99%﴿
* Data are the mean of at least 3 independent experiments. § The mean value of at least 6
independent experiments are given; 95% confidence interval and dose-response curves are
presented in the supporting information
Table 2. NMR Spectroscopic Data (DMSO-d6) for 5,6-dihydroxydamnacanthol (11)
position δH (J in Hz)
HMBC (2J, 3J)
C-1a, 2, 3, 4a, 10 Figure 1. Compounds isolated from the roots of Pentas longiflora

Figure 2
. Structures of known compounds isolated from the roots of Pentas lanceolata
Figure 3: Structure of 5, 6-Dihydroxydamnacanthol (11)


A Macroeconomic Analysis of Selected Economic Development Indicators in the Local Government Units in Specific Regions in the Philippines Ivy Benito-Lim The development concerns affected by economic indicators and being addressed are not new; they are in fact common to most Local Government Units (LGUs). The practices they employ, likewise, do not involve

18 the yukon old crow helicobacter pylori infection project

The Yukon Old Crow Helicobacter pylori Infection Project The First Report on the Prevalence and Epidemiology of Helicobacter pylori in Sander Veldhuyzen van Zanten, Laura Aplin, Amy L. Morse, John W. Morse, Monika M. Keelan, Janis Geary, Brendan Hanley, Diane M. Kirchgatter, Wendy Balsillie, Karen J. Dorji Dorji, Tashi D. Wangdi, Hoda M. Malaty, Kinley Wangchuk, Deki Yangzom, James