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Professor Dulcie Mulholland


Professor of Organic Chemistry
B.Sc, B.Sc(Hons), M.Sc, PhD(Natal)
+44 (0)1483 686751
02 AZ 03

Academic and research departments

Department of Chemistry.

Research

Research interests

My publications

Highlights

1. The Antiangiogenic Activity of Naturally-occurring and synthetic Homoisoflavonoids from the Hyacinthaceae (sensu APGII). SL Schwikkard, H Whitmore, RS. Sulaiman, K. Sishtla, T Shetty, H Basavarajappa, C Waller, A Alqhatani, L Frankemoelle, A Chapman, N Crouch, W Wetschnig, W Knirsch, J Andriantiana, MK Langat , TW Corson and D Mulholland, Journal of Natural Products, 2019, DOI: 10.1021/acs.jnatprod.8b00989

2. Application of Residual Dipolar Couplings and Selective Quantitative NOE to Establish the Structures of Tetranortriterpenoids from Xylocarpus rumphii (Meliaceae). W Waratchareeyakul, E. Hellemann, RR Gil, K Chantrapromma, MK Langat and DA Mulholland. 2017, Journal of Natural Products. 10.1021/acs.jnatprod.6b00906

3. Phytochemical investigations of three Rhodocodon (Hyacinthaceae sensu APG II) species. SL Schwikkard, A Alqahtani, W Knirsch, W Wetschnig, A Jaksevicius, EI Opara, MK Langat and DA Mulholland, 2017, Journal of Natural Products. DOI 10.1021/acs.jnatprod.6b00240

4. Diterpenoids from the roots of Croton dichogamus Pax. AHS Aldhaher, MK Langat. BM Ndunda, DK Chirchir, JO Midiwo, AW Njue, SL Schwikkard, M Carew, DA Mulholland. 2017, Phytochemistry, 144, 1-8.

5.Efficacy of extracts from eight economically important forestry species against downy mildew (Plasmopora viticola) and identification of active constituents. DA Mulholland*, MK Langat, EE James, DA Nawrot, L Tamm, B Thürig, Hans-Jakob Schärer, N Demidova, D Izotov, H Kleeberg, I Kleeberg, J Treutwein, S Cergel, H Hokkanen. 2017, Crop Protection, 102, 104-109.  

Publications

Sihra J, Langat M, Crouch N, Nuzillard J, Plainchont B, Mulholland D (2017) Novel triterpenoid derivatives from Eucomis bicolor Bak. (Hyacinthaceae: Hyacinthoideae),RSC Advances 7 pp. 15416-15427 Royal Society of Chemistry
The bulbs of Eucomis bicolor (Hyacinthoideae) yielded fourteen novel natural compounds, including (17S)-3-oxo-24,25,26,27,28-pentanorlanost-8-en-23,17±-olide, whose structure was determined using the Logic for Structure Determination Program, and nine novel lanosterol glycosides. Compounds were screened against the NCI-59 cancer cell panel but showed limited activity.
Reid KA, Maes J, Maes A, van Staden J, De Kimpe N, Mulholland DA, Verschaeve L (2006) Evaluation of the mutagenic and antimutagenic effects of South African plants, Journal of Ethnopharmacology 106 (1) pp. 44-50
Dichloromethane and 90% methanol extracts of 42 South African plants were screened for mutagenicity and antimutagenicity using the Salmonella/microsome mutagenicity assay (Ames) against Salmonella typhimurium TA98 and TA100 bacterial strains in the presence and absence of metabolic activator S9. The methanol extracts from whole plants of Helichrysum simillimum, Helichrysum herbaceum and Helichrysum rugulosum indicated mutagenicity. These are the first reported tests on the mutagenicity of Helichrysum species. Six species indicated antimutagenic properties, all in the presence of S9: methanol leaf extract of Bauhinia galpinii, and dichloromethane leaf extracts of Bauhinia galpinii, Clerodendrum myricoides, Datura stramonium, Buddleja saligna, Millettia sutherlandii and Sutherlandia frutescens. © 2005 Elsevier Ireland Ltd. All rights reserved.
Moodley B, Mulholland DA, Brookes HC (2012) The chemical oxidation of lignin found in Sappi Saiccor dissolving pulp mill effluent,WATER SA 38 (1) pp. 1-7 Water Research Commission
Claessens S, Naidoo D, Mulholland D, Verschaeve L, Van Staden J, De Kimpe N (2006) Synthesis of pyranonaphthoquinone antibiotics involving the phthalide annulation strategy, Synlett (4) pp. 621-623
The synthesis of the pyranonaphthoquinone antibiotic pentalongin was performed using the phthalide annulation strategy. Annulation of the cyanophthalide onto 6H-pyran-3-one resulted in a hongconin analogue, which upon further elaboration was converted into the natural product. © Georg Thieme Verlag Stuttgart.
Magadula JJ, Mulholland DA, Crouch NR (2008) Triterpenoids from Anthocleista grandiflora (Gentianaceae), NATURAL PRODUCT COMMUNICATIONS 3 (6) pp. 885-889 NATURAL PRODUCTS INC
Du Toit K, Elgorashi EE, Malan SF, Mulholland DA, Drewes SE, Van Staden J (2007) Antibacterial activity and QSAR of homoisoflavanones isolated from six Hyacinthaceae species, SOUTH AFRICAN JOURNAL OF BOTANY 73 (2) pp. 236-241 ELSEVIER SCIENCE BV
Dlova NC, Nevondo FT, Mwangi EM, Summers B, Tsoka-Gwegweni J, Martincigh BS, Mulholland DA (2013) Chemical analysis and in vitro UV-protection characteristics of clays traditionally used for sun protection in South Africa, PHOTODERMATOLOGY PHOTOIMMUNOLOGY & PHOTOMEDICINE 29 (3) pp. 164-169 WILEY-BLACKWELL
Randrianarivelojosia M, Mulholland DA, Mc Farland K (2005) Prenylated coumarins from Cedrelopsis longibracteata (Ptaeroxylaceae), Biochemical Systematics and Ecology 33 (3) pp. 301-304
Ndunda B, Langat MK, Mulholland DA, Eastman H, Jacob M, Khan S, Walker L, Muhammad I, Kerubo L, Midiwo J (2016) New ent-clerodane and abietane diterpenoids from the roots of Kenyan Croton megalocarpoides,Planta Medica 82 (11-12) pp. 1079-1086 Georg Thieme Verlag
The roots of the endangered medicinal plant Croton megalocarpoides (Euphorbiaceae) collected in Kenya has been investigated. Twenty two compounds has been isolated. Among them twelve new ¬ent-clerodane (1-12) and a new abietane (13) diterpenoid alongside the known crotocorylifuran (4a), two known abietane and four known ent-trachylobane diterpenoids, and the triterpenoids, lupeol and acetyl aleurotolic acid. The structures of the compounds were determined using NMR, HRMS and ECD. The isolated compounds have been evaluated against a series of microorganisms (fungal and bacteria) and also against Plasmodium falciparum, however no activity was observed.
Moodley N, Crouch NR, Mulholland DA, Slade D, Ferreira D (2006) 3-Benzyl-4-chromanones (homoisoflavanones) from bulbs of the ethnomedicinal geophyte Ledebouria revoluta (Hyacinthaceae), SOUTH AFRICAN JOURNAL OF BOTANY 72 (4) pp. 517-520 ELSEVIER SCIENCE BV
Crouch NR, du Toit K, Mulholland DA, Drewes SE (2006) Bufadienolides from bulbs of Urginea lydenburgensis (Hyacinthaceae : Urgineoideae), PHYTOCHEMISTRY 67 (19) pp. 2140-2145 PERGAMON-ELSEVIER SCIENCE LTD
Fusi F, Ferrara A, Koorbanally C, Crouch NR, Mulholland DA, Sgaragli G (2008) Vascular myorelaxing activity of isolates from South African Hyacinthaceae partly mediated by activation of soluble guanylyl cyclase in rat aortic ring preparations, JOURNAL OF PHARMACY AND PHARMACOLOGY 60 (4) pp. 489-497 PHARMACEUTICAL PRESS-ROYAL PHARMACEUTICAL SOC GREAT BRITIAN
Langat MK, Mulholland DA, Crouch NR (2008) New diterpenoids from Croton sylvaticus and Croton pseudopulchellus (Euphorbiaceae) and antiplasmodial screening of ent-kaurenoic acid, PLANTA MED
Obbo CJD, Makanga B, Mulholland DA, Coombes PH, Brun R (2013) Antiprotozoal activity of Khaya anthotheca, (Welv.) CDC a plant used by chimpanzees for self-medication,JOURNAL OF ETHNOPHARMACOLOGY 147 (1) pp. 220-223 ELSEVIER IRELAND LTD
Magadula JJ, Kapingu MC, Mbwambo ZH, Mulholland DA (2008) Secondary Metabolites from Teclea amanuensis (Rutaceae) from Tanzania, NATURAL PRODUCT COMMUNICATIONS 3 (10) pp. 1683-1686 NATURAL PRODUCTS INC
Koorbanally NA, Crouch NR, Harilal A, Pillay B, Mulholland DA (2006) Coincident isolation of a novel homoisoflavonoid from Resnova humifusa and Eucomis montana (Hyacinthoideae: Hyacinthaceae), Biochemical Systematics and Ecology 34 (2) pp. 114-118
We report on the first phytochemical investigation of a member of the African genus Resnova (Hyacinthoideae: Hyacinthaceae). From the dichloromethane extract of the bulbs of both Resnova humifusa and Eucomis montana (Hyacinthoideae: Hyacinthaceae) a novel 3-benzyl-4-chromanone homoisoflavonoid, 5,6-dimethoxy-7-hydroxy-3-(42-hydroxybenzyl)-4-chromanone, was isolated. A further 11 known homoisoflavonoids were also identified, the 12 in total presenting a clear biosynthetic sequence. Eight of the 12 compounds found were common to both species. © 2005 Elsevier Ltd. All rights reserved.
Essop AB, van Zyl RL, van Vuuren SF, Mulholland D, Viljoen AM (2008) The in vitro pharmacological activities of 12 South African Hermannia species, JOURNAL OF ETHNOPHARMACOLOGY 119 (3) pp. 615-619 ELSEVIER IRELAND LTD
Moodley B, Brookes HC, Mulholland DA (2011) The electro-oxidation of lignin in Sappi Saiccor dissolving pulp effluent.,Water SA 37 (1) Water Research Commission, SA
Electro-oxidation reactions using a nickel anode were carried out on the calcium-spent liquor effluent obtained from Sappi Saiccor (formerly South African Industrial Cellulose Corporation) dissolving pulp mill as well as on lignin- and lignan-type compounds previously identified in the effluent. Voltammograms were obtained for each solution in order to identify the oxidation potentials of the compounds to be electro-oxidised. Value-added products such as vanillin and syringaldehyde were identified in the electro-oxidised reaction mixtures using gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC). Profiles of the changes in concentration of these compounds were determined as a function of time with maximum concentrations reached within the first half hour. These findings are significant in that few electro-oxidation reactions have been carried out on the effluent of a pulp mill which uses the acid bisulphite pulping process and no results have previously been reported on the electro-oxidation of syringaldehyde. This study contributes to a further understanding of the electro-oxidation of lignin and is of value to the paper and pulp industry at large. Reduction of the organic content of the effluent by electro-oxidation was shown to be possible.
Mulholland DA, Langat MK, Crouch NR, Coley HM, Mutambi EM, Nuzillard J-M (2010) Cembranolides from the stem bark of the southern African medicinal plant, Croton gratissimus (Euphorbiaceae),PHYTOCHEMISTRY 71 (11-12) pp. 1381-1386 PERGAMON-ELSEVIER SCIENCE LTD
Mohammed AMA, Coombes PH, Crouch NR, Mulholland DA (2012) Garjasmine unequivocally isolated from the Rubiaceae,BIOCHEMICAL SYSTEMATICS AND ECOLOGY 44 pp. 109-111 PERGAMON-ELSEVIER SCIENCE LTD
Fusi F, Cavalli M, Mulholland D, Crouch N, Coombes P, Dawson G, Bova S, Sgaragli G, Saponara S (2010) Cardamonin Is a Bifunctional Vasodilator that Inhibits Ca(v)1.2 Current and Stimulates K(Ca)1.1 Current in Rat Tail Artery Myocytes, JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 332 (2) pp. 531-540 AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS
The bulbs of Ledebouria socialis (Hyacinthaceae) yielded the benzocyclobutene homoisoflavonoid, (R)-2',5-dihydroxy-3',4',7-trimethoxyspiro{2H-1-benzopyran-3-(4H)-9-bicyclo[4.2.0]octa[1,3,5]triene}-4-one, socialinone (1). Ledebouria ovatifolia yielded (2µ,3R)-2,5-dihydroxy-7-methoxyspiro[2H-1-benzopyran-3(4H), 5'(6'H)-cyclobuta[f][1,3]benzodioxol]-4-one (2) and the homoisoflavanone, (E)-3-(3',4'-dihydroxybenzylidene)-5,7-dihydroxychroman-4-one, ovatifolionone (5), the dihydrochalcone, 4,4'-dihydroxy-2',6'-dimethoxydihydrochalcone (3), and xanthone, 1,6-dihydroxy-2,3,5-trimethoxy-8-methyl-9H-xanthen-9-one (4) along with 21 known compounds. Structures were determined using spectroscopic techniques. The anti-inflammatory activities of the homoisoflavonoids and xanthones isolated were evaluated against cyclooxygenase-1 and -2 isoenzymes. (R)-3-(3',4'-Dihydroxybenzyl)-7-hydroxy-5-methoxychroman-4-one (7), (E)-3-(3',4'-dihydroxybenzylidene)-7-hydroxy-5-methoxychroman-4-one (10), 1,3,6-trihydroxy-2-methoxy-8-methylxanthen-9-one (6) and ovatifolionone acetate (5Ac) exhibited significant activity against cyclooxygenase-2 at
Coombes PH, Mwangi EM, Peters BK, Crouch NR, Mulholland DA (2009) The cyclobuta[b]quinoline alkaloid cyclomegistine from Teclea gerrardii I.Verd. (Toddalioideae: Rutaceae),BIOCHEMICAL SYSTEMATICS AND ECOLOGY 37 (4) pp. 494-496 PERGAMON-ELSEVIER SCIENCE LTD
Coombes PH, Mulholland DA, Randrianarivelojosia M (2008) Vilasinin limonoids from Malleastrum antsingyense J.F. Leroy (Meliodeae : Meliaceae),BIOCHEMICAL SYSTEMATICS AND ECOLOGY 36 (1) pp. 74-76 PERGAMON-ELSEVIER SCIENCE LTD
Tunbridge CP, Crouch NR, Thumser AE, Mulholland DA (2012) COX-2 SPECIFIC INHIBITORS FROM LEDEBOURIA OVATIFOLIA AND LEDEBOURIA SOCIALIS (HYACINTHACEAE:HYACINTHOIDEAE), PHARMACEUTICAL BIOLOGY 50 (5) pp. 569-569
Tarus PK, Coombes PH, Crouch NR, Mulholland DA, Moodley B (2005) Furoquinoline alkaloids from the southern African Rutaceae Teclea natalensis., Phytochemistry 66 (6) pp. 703-706
The chloroform and ethyl acetate extracts of the leaves of Teclea natalensis have yielded two furoquinoline alkaloids, 6-[(2,3-epoxy-3-methylbutyl)oxy]-4,7-dimethoxyfuro[2,3-b]quinoline and 4,7-dimethoxy-6-[(3-methyl-2-butenyl)oxy]furo[2,3-b]quinoline, and the known alkaloids 4,7-dimethoxy-8-[(3-methyl-2-butenyl)oxy]furo[2,3-b]quinoline, flindersiamine and dictamnine.
Waratchareeyakul W, Rocco A, Fusi F, Schwikkard SL, Knirsch W, Mulholland DA (2015) Vasorelaxant studies on stilbenoids and phenanthrene derivatives from Brasiliorchis porphyrostele Rchb.f. (Orchidaceae), PLANTA MEDICA 81 (16) pp. 1418-1418 GEORG THIEME VERLAG KG
Schwikkard S, James EE, Mulholland DA, Knirsch W, Corson T, Opara E, Jaksevicius A (2015) Novel, bioactive homoisoflavonoids from a Madagascan Rhodocodon species (Hyacinthaceae, sensu APG II), PLANTA MEDICA 81 (16) pp. 1451-1451 GEORG THIEME VERLAG KG
Tsamo A, Langat MK, Nkounga P, Kamdem Waffo AF, Nkengfack AE, Mulholland DA (2013) Limonoids from the West African Trichilia welwitschii (Meliaceae),Biochemical Systematics and Ecology 50 pp. 368-370
The seeds of Trichilia welwitschii C.DC. (Meliaceae) yielded three limonoids, dregeanin DM4 (1), reported here from a natural source for the first time, the known rohituka 3 (2) and trichilia lactone D5 (3). The bark yielded 28,29-dinorcycloart-24-ene-3,4,6-triol (4), sitosterol-3-O-²-d-glucoside, 4-hydroxy-N-methyl-l-proline, stigmasterol and sitosterol. © 2013 Elsevier Ltd.
Mohammed AMA, Coombes PH, Crouch NR, Mulholland DA (2009) Non-volatile isolates of two Heteropyxis species: A first chemotaxonomic assessment of subfamily Psiloxyloideae (Myrtaceae), BIOCHEMICAL SYSTEMATICS AND ECOLOGY 37 (3) pp. 241-243 PERGAMON-ELSEVIER SCIENCE LTD
Reid KA, Jäger AK, Light ME, Mulholland DA, Van Staden J (2005) Phytochemical and pharmacological screening of Sterculiaceae species and isolation of antibacterial compounds., J Ethnopharmacol 97 (2) pp. 285-291
Little previous phytochemical investigation has been conducted on South African Sterculiaceae species used in traditional medicine. In this study, five species, varying in growth type (small herbs, shrubs and large trees) and traditional usage were investigated. The species screened were Cola greenwayi Brenan, Cola natalensis Oliv., Dombeya burgessiae Gerr. ex Harv., Dombeya cymosa Harv. and Hermannia depressa N.E.Br. Extracts were screened for alkaloids, cardiac glycosides, cyanogenic glycosides, saponins and tannins. The probable presence of bufadienolides in the leaf material of Dombeya burgessiae and Dombeya cymosa was determined. Alkaloids, cyanogenic glycosides and saponins were absent in all the plant material investigated. Tannins were detected in the leaf extract of Cola greenwayi and in the leaves, stems and roots of Hermannia depressa. Extracts were screened for anti-inflammatory and antibacterial activity using the cyclooxygenase-1 (COX-1) inhibition assay and the microdilution antibacterial assay. The ethanol and dichloromethane extracts of Cola greenwayi, Dombeya burgessiae and Dombeya cymosa, and the dichloromethane extracts of Hermannia depressa showed the highest levels of COX-1 inhibition. It is possible that the high levels observed may be due to the presence of tannins in some of the extracts. Generally, all the aqueous extracts exhibited low activity. Similarly, no antibacterial activity was observed with the aqueous extracts, although some mild activity was exhibited with some of the ethanol and ethyl acetate extracts. Following the general phytochemical and pharmacological screening, extracts showing antibacterial activity were further purified using bioassay-guided fractionation. Dombeya rotundifolia (Hochst.) Planch., which was screened in a previous study, was also included in the isolation of active compounds. A bioautographic assay, using Staphylococcus aureus, was used to detect the presence of the antibacterial compounds. These were isolated and identified as fatty acids.
Mulholland DA, Crouch NR, Koorbanally C, Moodley N, Pohl T (2006) Infraspecific chemical variation in Scilla zebrina (Hyacinthaceae), BIOCHEMICAL SYSTEMATICS AND ECOLOGY 34 (3) pp. 251-255 PERGAMON-ELSEVIER SCIENCE LTD
Schwikkard SL, Mulholland DA (2014) Useful methods for targeted plant selection in the discovery of potential new drug candidates.,Planta medica 80 (14) pp. 1154-1160
The efficient and effective selection of appropriate plants for investigative purposes in a drug discovery program is of crucial importance for a successful outcome. A variety of approaches have been used by researchers with varying levels of success. A variety of different approaches to plant selection are discussed, including the ethnomedicinal approach, some ecological approaches, and the use of combinatorial and computational methodologies.
Mulholland DA, Langlois A, Randrianarivelojosia M, Derat E, Nuzillard JM (2006) The structural elucidation of a novel iridoid derivative from Tachiadenus longiflorus (Gentianaceae) using the LSD programme and quantum chemical computations, PHYTOCHEMICAL ANALYSIS 17 (2) pp. 87-90 JOHN WILEY & SONS LTD
Sihra JK, Thumser AE, Langat MK, Crouch NR, Mulholland DA (2015) Constituents of Bulbs of three Species of the Hyacinthaceae (Hyacinthoideae): Eucomis vandermerwei, E. zambesiaca and Resnova humifusa, NATURAL PRODUCT COMMUNICATIONS 10 (7) pp. 1207-1209 NATURAL PRODUCTS INC
Tembu VJ, Langat MK, Crouch NR, Mulholland DA (2014) Use of Circular Dichroism to Determine the Absolute Configuration of a Pimarane Diterpenoid from the Southern African Sclerocroton integerrimus (Euphorbiaceae), NATURAL PRODUCT COMMUNICATIONS 9 (8) pp. 1131-1133 NATURAL PRODUCTS INC
Mulholland DA (2005) The future of ethnopharmacology: a southern African perspective., J Ethnopharmacol 100 (1-2) pp. 124-126
The traditional use of medicinal plants in southern Africa is widespread with over 80% of South African households using medicinal plants. A mushrooming of research into the study of medicinal plants in the developing world has occurred but most of Africa is lagging behind in research output despite long traditions of medicinal plant usage. A considerable investment in research infrastructure is required.
Coombes PH, Naidoo D, Mulholland DA, Randrianarivelojosia M (2005) Quassinoids from the leaves of the Madagascan Simaroubaceae Samadera madagascariensis, PHYTOCHEMISTRY 66 (23) pp. 2734-2739 PERGAMON-ELSEVIER SCIENCE LTD
Ismail F, Mulholland DA, Marsh JJ (2005) An analysis of the water soluble components of Sappi Saiccor's effluent streams, Water SA 31 (4) pp. 569-574
Sappi Saiccor is a pulp mill that produces high-grade chemical cellulose (dissolving pulp) from predominantly hardwood timber and is currently the world's largest manufacturer of this type of pulp. Attempts to isolate pure lignosulphonates were unsuccessful; however, an acid hydrolysis of the aqueous portion of the calcium effluent stream yielded a range of organic compounds. These included lignans, lignin-type precursors as well as small quantities of vanillin and syringaldehyde. The structures of these compounds were determined using NMR spectroscopic and mass spectrometric techniques.
Crouch NR, Mulholland DA, Chetty J (2005) Lupane triterpenoid and alkaloid isolates from Cyrtanthus breviflorus (Amaryllidaceae), South African Journal of Botany 71 (1) pp. 104-106
Cyrtanthus breviflorus Harv. is a highly variable bulbous taxon from both moist and dry grasslands of the eastern seaboard region of southern Africa. Amongst at least 42 ethnomedicinal regional amaryllids it is the only species reported to treat intestinal worms. The current investigation, the first to phytochemically profile C. breviflorus, has determined the presence in ethanolic bulb extracts of the four known isoquinoline alkaloids: haemanthamine, crinamine hydrochloride, lycorine and tazettine. From the hexane bulb extract five known lupane triterpenoids were isolated: lupeol, lupenone, glochidone, 3², 27-dihydroxylup-20(29)-ene and betulinaldehyde. Copyright © NISC Pty Ltd.
Koorbanally C, Mulholland DA, Crouch NR (2006) Norlignans and homoisoflavanones from two south African Drimiopsis species (Hyacinthaceae : Hyacinthoideae), BIOCHEMICAL SYSTEMATICS AND ECOLOGY 34 (7) pp. 588-592 PERGAMON-ELSEVIER SCIENCE LTD
Kamdem Waffo AF, Coombes PH, Crouch NR, Mulholland DA, El Amin SMM, Smith PJ (2007) Acridone and furoquinoline alkaloids from Teclea gerrardii (Rutaceae : Toddalioideae) of southern Africa, PHYTOCHEMISTRY 68 (5) pp. 663-667 PERGAMON-ELSEVIER SCIENCE LTD
Langat MK, Crouch NR, Pohjala L, Tammela P, Smith PJ, Mulholland DA (2012) Ent-kauren-19-oic acid derivatives from the stem bark of Croton pseudopulchellus Pax, Phytochemistry Letters 5 (3) pp. 414-418
Two new ent-kauren-19-oic acid derivatives, ent-14S*-hydroxykaur-16- en-19-oic acid and ent-14S*,17-dihydroxykaur-15-en-19-oic acid together with eleven known compounds ent-kaur-16-en-19-oic acid, ent-kaur-16-en-19-al, ent-12²-hydroxykaur-16-en-19-oic acid, ent-12²-acetoxykaur-16-en-19- oic acid, 8R,13R-epoxylabd-14-ene, eudesm-4(15)-ene-1²,6±-diol, (-)-7-epivaleran-4-one, germacra-4(15), 5E,10(14)-trien-9²-ol, acetyl aleuritolic acid, ²-amyrin, and stigmasterol were isolated from the stem bark of Croton pseudopulchellus (Euphorbiaceae). Structures were determined using spectroscopic techniques. Ent-14S*-hydroxykaur-16-en-19-oic acid, ent-kaur-16-en-19-oic acid, ent-12²-hydroxykaur-16-en-19-oic acid, ent-12²-acetoxykaur-16-en-19-oic acid and 8R,13R-epoxylabd-14-ene were tested for their effects on Semliki Forest virus replication and for cytotoxicity against human liver tumour cells (Huh-7 strain) but were found to be inactive. Ent-kaur-16-en-19-oic acid, the major constituent, showed weak activity against the Plasmodium falciparum (CQS) D10 strain. © 2012 Phytochemical Society of Europe.
Douwes E, Crouch NR, Edwards TJ, Mulholland DA (2008) Regression analyses of southern African ethnomedicinal plants: informing the targeted selection of bioprospecting and pharmacological screening subjects,Journal of Ethnopharmacology 119 (3) pp. 356-364
Ethnopharmacological relevance: Regression analyses of local medicinal floras are considered potentially useful when prioritising candidate plant taxa for pharmacological/bioprospecting investigations. Aim of the study: To identify plant orders and subsequently families within the highly diverse ethnomedicinal flora of southern Africa, towards which biases by traditional healers are demonstrable. Taxa so identified can subsequently be weighted appropriately in semi-quantitative selection systems. Methodology: Plant data sourced from the SANBI MedList database, the most comprehensive inventory of ethnomedicinal plants for the Flora of southern Africa region were grouped by order. A least squares regression analysis was applied to test the null hypothesis that the use of these plants by traditional healers is strictly random. Of 'hot' orders subsequently identified, characteristics of taxa therein were assessed to better determine the roles played by (i) growth forms, and (ii) inherent chemical diversity, in plant selections by ethnomedicinal practitioners. Results: Analyses identified seven principally 'hot' plant orders (Malpigiales, Fabales, Gentianales, Asteraceae, Solanales, Malvales and Sapindales) and 'hot' families therein from a total of 55 regional ethnomedicinal orders. Five 'cold' ethnomedicinal orders (Rosales, Proteales, Poales, Asparagales and Caryophyllales) were shown to be significantly less represented in the medicinal flora than predicted. No clear growth form preferences were identified across orders. The presence of highly diverse bioactives was evident in the 'hottest' plant families from 'hot' plant orders. Conclusions: These 12 outliers identified by the regression analyses allowed for the falsification of the null hypothesis. Indications are that 'hot' taxa are selected traditionally on the basis of bioactivity, which is reflected in chemical diversity. © 2008 Elsevier Ireland Ltd. All rights reserved.
Chukwujekwu JC, Coombes PH, Mulholland DA, van Staden J (2006) Emodin, an antibacterial anthraquinone from the roots of Cassia occidentalis, South African Journal of Botany 72 (2) pp. 295-297
The antibacterial activity of the ethanolic root extract of Cassia occidentalis was examined. A biologically active component was isolated and identified as emodin by spectroscopic analysis. The bioactive Minimum Inhibitory Concentration (MIC) values of emodin were 7.8 × 10- 3 and 3.9 × 10- 3 mg ml- 1 against Bacillus subtilis and Staphylococcus aureus, respectively. It was not active against two Gram-negative bacteria (Klebsiella pneumoniae and Escherichia coli) at the highest concentration (5.0 × 10- 1 mg ml- 1) tested. © 2005 Elsevier B.V. All rights reserved.
Kapingu MC, Magadula JJ, Mbwambo ZH, Mulholland DA (2008) Puguflavanones A and B; Prenylated flavanones from Baphia puguensis, NATURAL PRODUCT COMMUNICATIONS 3 (5) pp. 749-753 NATURAL PRODUCTS INC
Nawrot DA, Randrianarivelojosia M, Langat MK, Mulholland DA (2013) Coumarins from the Malagasy Cedrelopsis rakotozafyi Cheek and Lescot (Rutaceae),Biochemical Systematics and Ecology 50 pp. 452-454
A phytochemical analysis of the bark of the Malagasy Cedrelopsis rakotozafyi Cheek and Lescot (Rutaceae) yielded the novel 8-hydroxy-7-methoxy-6-(2. R-hydroxy-3-methylbut-3-enoxy)-2H-1-benzopyran-2-one, and 7-hydroxy-6-(2. R-hydroxy-3-methylbut-3-enoxy)-2H-1-benzopyran-2-one, 5,6,7-trimethoxy-2H-1-benzopyran-2-one, scoparone, scopoletin, lupeol and ²-amyrin. The placement of Cedrelopsis within the Rutaceae is supported phytochemically by the typically Rutaceous coumarins isolated. © 2013 Elsevier Ltd.
Mulholland DA, Langat MK, Crouch NR, Coley HM, Mutambi EM, Nuzillard JM (2010) Cembranolides from the stem bark of the southern African medicinal plant, Croton gratissimus (Euphorbiaceae), Phytochemistry 71 (11-12) pp. 1381-1386
The stem bark of Croton gratissimus (Euphorbiaceae) yielded four cembranolides, including the first reported example of a 2,12-cyclocembranolide, (+)-[1R*,2S*,7S*,8S*,12R*]-7,8-epoxy-2,12- cyclocembra-3E,10Z-dien-20,10-olide, whose structure was confirmed by means of single crystal X-ray analysis. This compound showed moderate activity against the PEO1 and PEO1TaxR ovarian cancer cell lines. © 2010 Elsevier Ltd. All rights reserved.
Mulholland DA, Schwikkard SL, Crouch NR (2013) The chemistry and biological activity of the Hyacinthaceae., Nat Prod Rep 30 (9) pp. 1165-1210 Royal Society of Chemistry
Covering: 1914 to 2012The Hyacinthaceae (sensu APGII), with approximately 900 species in about 70 genera, can be divided into three main subfamilies, the Hyacinthoideae, the Urgineoideae and the Ornithogaloideae, with a small fourth subfamily the Oziroëoideae, restricted to South America. The plants included in this family have long been used in traditional medicine for a wide range of medicinal applications. This, together with some significant toxicity to livestock has led to the chemical composition of many of the species being investigated. The compounds found are, for the most part, subfamily-restricted, with homoisoflavanones and spirocyclic nortriterpenoids characterising the Hyacinthoideae, bufadienolides characterising the Urgineoideae, and cardenolides and steroidal glycosides characterising the Ornithogaloideae. The phytochemical profiles of 38 genera of the Hyacinthaceae will be discussed as well as any biological activity associated with both crude extracts and compounds isolated. The Hyacinthaceae of southern Africa were last reviewed in 2000 (T. S. Pohl, N. R. Crouch and D. A. Mulholland, Curr. Org. Chem., 2000, 4, 1287-1324; ); the current contribution considers the family at a global level.
Cheplogoi PK, Mulholland DA, Coombes PH, Randrianarivelojosia M (2008) An azole, an amide and a limonoid from Vepris uguenensis (Rutaceae),PHYTOCHEMISTRY 69 (6) pp. 1384-1388 PERGAMON-ELSEVIER SCIENCE LTD
Naidoo D, Coombes PH, Mulholland DA, Crouch NR, van den Bergh AJ (2005) N-Substituted acridone alkaloids from Toddaliopsis bremekampii (Rutaceae: Toddalioideae) of south-central Africa., Phytochemistry 66 (14) pp. 1724-1728
Toddaliopsins A-D, four novel 1,2,3-trioxygenated acridone alkaloids, have been isolated from the leaves of Toddaliopsis bremekampii. Toddaliopsins B-D are the first reported acridone alkaloids with substituted N-methyl groups, in the light of which the chemotaxonomic relationship of Toddaliopsis and Vepris is discussed. Toddaliopsin C possesses moderate anti-inflammatory activity, which may be related to the hydroxy group present at C-1.
Moodley N, Neil RC, Mulholland DA (2007) Bufadienolides from drimia macrocentra and urginea riparia (Hyacinthaceae: urgineoideae), PHYTOCHEMISTRY 68 (19) pp. 2415-2419 PERGAMON-ELSEVIER SCIENCE LTD
Langat MK, Mulholland DA, Langat MK, Crouch NR, Mulholland DA, Crouch NR, Pohjala L, Tammela P, Pohjala L, Smith PJ (2012) Ent-kauren-19-oic acid derivatives from the stem bark of Croton pseudopulchellus Pax,Phytochemistry Letters 5 (3) pp. 414-418 Elsevier
Two new ent-kauren-19-oic acid derivatives, ent-14S*-hydroxykaur-16-en-19-oic acid and ent-14S*,17-dihydroxykaur-15-en-19-oic acid together with eleven known compounds ent-kaur-16-en-19-oic acid, ent-kaur-16-en-19-al, ent-12²-hydroxykaur-16-en-19-oic acid, ent-12²-acetoxykaur-16-en-19-oic acid, 8R,13R-epoxylabd-14-ene, eudesm-4(15)-ene-1²,6±-diol, (-)-7-epivaleran-4-one, germacra-4(15), 5E,10(14)-trien-9²-ol, acetyl aleuritolic acid, ²-amyrin, and stigmasterol were isolated from the stem bark of Croton pseudopulchellus (Euphorbiaceae). Structures were determined using spectroscopic techniques. Ent-14S*-hydroxykaur-16-en-19-oic acid, ent-kaur-16-en-19-oic acid, ent-12²-hydroxykaur-16-en-19-oic acid, ent-12²-acetoxykaur-16-en-19-oic acid and 8R,13R-epoxylabd-14-ene were tested for their effects on Semliki Forest virus replication and for cytotoxicity against human liver tumour cells (Huh-7 strain) but were found to be inactive. Ent-kaur-16-en-19-oic acid, the major constituent, showed weak activity against the Plasmodium falciparum (CQS) D10 strain. © 2012 Phytochemical Society of Europe.
Mulholland DA, McFarland K, Randrianarivelojosia M (2006) Sesquiterpenoid derivatives from Cipadessa boiviniana (Meliaceae), BIOCHEMICAL SYSTEMATICS AND ECOLOGY 34 (4) pp. 365-369 PERGAMON-ELSEVIER SCIENCE LTD
Nawrot DA, Langat MK, Mulholland DA (2012) CHEMICAL CONSTITUENTS OF EAST EUROPEAN FOREST SPECIES, PHARMACEUTICAL BIOLOGY 50 (5) pp. 588-588
Waffo AFK, Coombes PH, Mulholland DA, Nkengfack AE, Fomum ZT (2006) Flavones and isoflavones from the west African Fabaceae Erythrina vogelii, PHYTOCHEMISTRY 67 (5) pp. 459-463 PERGAMON-ELSEVIER SCIENCE LTD
Koorbanally C, Mulholland DA, Crouch NR (2005) A novel 3-hydroxy-3-benzyl-4-chromanone-type homoisoflavonoid from Albuca fastigiata (Ornithogaloideae: Hyacinthaceae), Biochemical Systematics and Ecology 33 (5) pp. 545-549
Waffo AFK, Mulholland D, Wansi JD, Mbaze LM, Powo R, Mpondo TN, Fomum ZT, Konig W, Nkengfack AE (2006) Afzeliixanthones A and B, two new prenylated xanthones from Garcinia afzelii ENGL. (Guttiferae), CHEMICAL & PHARMACEUTICAL BULLETIN 54 (4) pp. 448-451 PHARMACEUTICAL SOC JAPAN
Coombes PH, Mulholland DA, Randrianarivelojosia M (2005) Mexicanolide limonoids from the Madagascan Meliaceae Quivisia papinae., Phytochemistry 66 (10) pp. 1100-1107
An investigation of the seeds of the Madagascan Meliaceae Quivisia papinae has yielded five mexicanolide group limonoids, together with two known mexicanolide limonoids and two known triterpenoids. Quivisianolide A 9 possesses a hitherto unreported 9alpha,11alpha-epoxide ring, quivisianolide B 10 the corresponding delta(9(11)) double bond, and quivisianone 11 is a 17-keto seco-ring D compound.
Langlois A, Mulholland DA, Duncan GD, Crouch NR, Edwards TJ (2005) A novel 3-benzylchromone from the South African Lachenalia rubida (Hyacinthaceae), Biochemical Systematics and Ecology 33 (9) pp. 961-966
Koorbanally C, Mulholland DA, Crouch NR (2005) Eudesmane-type sesquiterpenoids from Urginea epigea (Urgineoideae; Hyacinthaceae), Biochemical Systematics and Ecology 33 (3) pp. 295-299
Vazquez R, Riveiro ME, Vermeulen M, Mondillo C, Coombes PH, Crouch NR, Ismail F, Mulholland DA, Baldi A, Shayo C, Davio C (2012) Toddaculin, a natural coumarin from Toddalia asiatica, induces differentiation and apoptosis in U-937 leukemic cells,PHYTOMEDICINE 19 (8-9) pp. 737-746 ELSEVIER GMBH, URBAN & FISCHER VERLAG
du Toit K, Elgorashi EE, Malan SF, Drewes SE, van Staden J, Crouch NR, Mulholland DA (2005) Anti-inflammatory activity and QSAR studies of compounds isolated from Hyacinthaceae species and Tachiadenus longiflorus Griseb. (Gentianaceae)., Bioorg Med Chem 13 (7) pp. 2561-2568
Twenty-two homoisoflavanones and structurally related compounds isolated from plants were screened for anti-inflammatory activity. Seventeen compounds were isolated from southern African Hyacinthaceae species, one from the Madagascan gentian Tachiadenus longiflorus Griseb. and four were of synthetic origin. Inhibition of prostaglandin synthesis in cell microsomal fractions was first evaluated, followed by screening for specific inhibition of isolated cyclooxygenase enzymes (COX-1 and COX-2). Six homoisoflavanone and structurally related compounds showed significantly high levels of anti-inflammatory activity in the microsomal fraction assay. Only one compound exhibited a high level of anti-inflammatory activity in the COX-1 enzyme assay and no significant activity was detected in the COX-2 enzyme assay. Biological screening was followed by a computer-based quantitative structure-activity relationship (QSAR) study. The physicochemical descriptors: strain energy, heat of formation, volume, surface area, aqueous phase energy, dipole moment, enthalpy, entropy, molar refractivity, parachor, density, refractive index, surface tension, polarizability, logP, Van der Waals interaction energy, Coulombic interaction energy and nonbonded interaction energy were used to characterize the structures of the homoisoflavanones and structurally related compounds. This study produced three equations with significant prediction values for the anti-inflammatory activity of the compounds investigated. The derived models also provided valuable parameter guidelines for those properties influencing the anti-inflammatory activity of the studied compounds.
Koorbanally C, Mulholland DA, Crouch NR (2005) A novel homoisoflavonoid from Drimia delagoensis (Urgineoideae: Hyacinthaceae), Biochemical Systematics and Ecology 33 (7) pp. 743-748
Langat MK, Crouch NR, Smith PJ, Mulholland DA (2011) Cembranolides from the Leaves of Croton gratissimus.,J Nat Prod 74 (11) pp. 2349-2355 American Chemical Society
Ten new cembranolides, (-)-(1R*,4R*,10R*)-4-methoxycembra-2E,7E,11Z-trien-20,10-olide (1), (-)-(1S*,4R*,10R*)-1-hydroxy-4-methoxycembra-2E,7E,11Z-trien-20,10-olide (2), (-)-(1S*,4S*,10R*)-1,4-dihydroxycembra-2E,7E,11Z-trien-20,10-olide (3), (-)-(1S*,4S*,10R*)-1,4-dihydroxycembra-2E,7E,11Z-trien-20,10-olide (4), (+)-(10R*)-cembra-1E,3E,7E,11Z,16-pentaen-20,10-olide (5), (+)-(10R*)-cembra-1Z,3Z,7E,11Z,15-pentaen-20,10-olide (6), (+)-(5R*,10R*)-5-methoxycembra-1E,3E,7E,11Z,15-pentaen-20,10-olide (7), (+)-(1S*,4S*,7R*,10R*)-1,4,7-trihydroxycembra-2E,8(19),11Z-trien-20,10-olide (8), (-)-(1S*,4S*,7S*,10R*)-1,4,7-trihydroxycembra-2E,8(19),11Z-trien-20,10-olide (9), and (+)-(1S*,4R*,8S*,10R*)-1,4,8-trihydroxycembra-2E,6E,11Z-trien-20,10-olide (10), together with six known compounds, lupeol, 4(15)-eudesmene-1²,6±-diol, ±-glutinol, 24-ethylcholesta-4,22-dien-3-one, (+)-(1R*,10R*)-cembra-2E,4E,7E,11Z-tetraen-20,10-olide, and (+)-(1R*,4S*,10R*)-4-hydroxycembra-2E,7E,11Z-trien-20,10-olide (4a), have been isolated from the leaves of Croton gratissimus. The acetyl derivatives of 8 and 4a were evaluated against a chloroquine-sensitive strain of Plasmodium falciparum (D10).
Mulholland DA, Crouch NR, Randrianarivelojosia M (2008) Phytochemistry and chemotaxonomy of the Madagascan Cedrelopsis, PLANTA MED 74 (9) pp. 914-914 GEORG THIEME VERLAG KG
Tarus PK, Coombes PH, Crouch NR, Mulholland DA (2006) Benzo[c]phenanthridine alkaloids from stem bark of the Forest Knobwood, Zanthoxylum davyi (Rutaceae), South African Journal of Botany 72 (4) pp. 555-558
The stem bark of the southern African ethnomedicinal tree Zanthoxylum davyi (Rutaceae) has yielded five benzo[c]phenanthridine alkaloids, chelerythrine, dihydrochelerythrine, bocconoline, 6-hydroxydihydrochelerythrine and 6-methoxy-7-demethyldihydrochelerythrine, together with 4-methoxy-1-methyl-2(1H)-quinolinone and the uncommon lignan meso-sesamin. The chemotaxonomic and ethnopharmacological relevance of these compounds is discussed, particularly that of the well-documented antimicrobial and anti-inflammatory chelerythrine. © 2006 Elsevier B.V. All rights reserved.
Mulholland DA, Mohammed AMA, Coombes PH, Haque S, Pohjala LL, Tammela PSM, Crouch NR (2011) Triterpenoid Acids and Lactones from the Leaves of Fadogia tetraquetra var. tetraquetra (Rubiaceae), NATURAL PRODUCT COMMUNICATIONS 6 (11) pp. 1573-1576
Ndlebe VJ, Crouch NR, Mulholland DA (2008) Triterpenoids from the African tree Phyllanthus polyanthus,PHYTOCHEMISTRY LETTERS 1 (1) pp. 11-17 ELSEVIER SCIENCE BV
Eldeen IM, Elgorashi EE, Mulholland DA, van Staden J (2006) Anolignan B: a bioactive compound from the roots of Terminalia sericea., J Ethnopharmacol 103 (1) pp. 135-138
Antibacterial bioassay-guided fractionation of an ethyl acetate root extract of Terminalia sericea led to the isolation of anolignan B. The isolated compound was further tested for anti-inflammatory activity using the cyclooxygenase enzyme assays (COX-1 and COX-2) and for potential mutagenic effects using the Ames test. In the antibacterial test, anolignan B showed activity against both Gram-positive and Gram-negative bacteria. The minimum inhibitory concentration values obtained (MIC) ranged from 3.8 microg/ml against Bacillus subtilis (Gram-positive) to 31 microg/ml against Escherichia coli (Gram-negative). In the anti-inflammatory assays, anolignan B showed activity against both COX-1 (IC(50) = 1.5 mM) and COX-2 (IC(50) = 7.5 mM) enzymes. No potential mutagenic effects were observed in the Salmonella microsome assay (TA98). Isolation of anolignan B from Terminalia sericea as well as the antibacterial and anti-inflammatory activities observed in this study has not been reported previously.
Penali L, Mulholland DA, Tano KD, Cheplogoi PK, Randrianarivelojosia M (2007) Low antiplasmodial activity of alkaloids and amides from the stem bark of Zanthoxylum rubescens (Rutaceae), PARASITE-JOURNAL DE LA SOCIETE FRANCAISE DE PARASITOLOGIE 14 (2) pp. 161-164 PRINCEPS EDITIONS
Tarus PK, Coombes PH, Crouch NR, Mulholland DA (2006) Benzo[c]phenanthridine alkaloids from stem bark of the Forest Knobwood, Zanthoxylum davyi (Rutaceae), SOUTH AFRICAN JOURNAL OF BOTANY 72 (4) pp. 555-558 ELSEVIER SCIENCE BV
Randrianarivelojosia M, Langlois A, Mulholland DA (2006) Investigations of the Malagasy species Tachiadenus longiflorus Grisebach (Gentianaceae): Linking chemical finding and traditional usage, JOURNAL OF ETHNOPHARMACOLOGY 105 (3) pp. 456-458 ELSEVIER IRELAND LTD
Tunbridge CP, Crouch NR, Thumser AE, Mulholland DA (2012) COX-2 SPECIFIC INHIBITION FROM NATURAL PRODUCT (E)-HINOKIRESINOL AND A FACILE SYNTHESIS OF 3-VINYLPHENYLINDANES, PHARMACEUTICAL BIOLOGY 50 (5) pp. 649-649
Crouch NR, Langlois A, Mulholland DA, Nair JJ (2005) A novel alkylamide from the leaves of Acmella caulirhiza (Asteraceae), a traditional surface analgesic, South African Journal of Botany 71 (2) pp. 228-230
Dried leaves of the widespread African ethnomedicinal herb Acmella caulirhiza (Asteraceae) have been phytochemically characterised. The hexane extract yielded two unsaturated alkylamides, spilanthol and a novel compound tentatively identified as N-isobutylnona-2E, 4E-dien-8-ynamide based on 1H, 13C, COSY and HETCOR spectra. The occurrence of these amides supports the validation of documented traditional usage patterns. Copyright © NISC Pty Ltd.
Ezra L, Waratchareeyakul W, Chantrapromma K, Chantrapromma S, Langat MK, Schwikkard SL, Mulholland DA (2015) Chemical constituents from Xylocarpus rumphii (Meliaceae), PLANTA MEDICA 81 (16) pp. 1462-1462 GEORG THIEME VERLAG KG
Munro OQ, du Toit K, Drewes SE, Crouch NR, Mulholland DA (2006) Experimental and theoretical studies of a naturally occurring non-oligomeric steroidal supramolecular zipper, NEW JOURNAL OF CHEMISTRY 30 (2) pp. 197-207 ROYAL SOC CHEMISTRY
Chukwujekwu JC, Smith P, Coombes PH, Mulholland DA, van Staden J (2005) Antiplasmodial diterpenoid from the leaves of Hyptis suaveolens., J Ethnopharmacol 102 (2) pp. 295-297
Bioactivity-guided fractionation of the petroleum ether extract of the leaves of Hyptis suaveolens, widely used in Traditional Medicine, has led to the isolation of an abietane-type diterpenoid endoperoxide, 13 alpha-epi-dioxiabiet-8(14)-en-18-ol, displaying high antiplasmodial activity (IC(50) 0.1 microg/ml).
Mulholland DA, Mwangi E, Dlova N, Plant N, Crouch N, Coombes P (2013) Non-toxic melanin production inhibitors from Garcinia livingstonei (Clusiaceae),JOURNAL OF ETHNOPHARMACOLOGY 149 (2) pp. 570-575 ELSEVIER IRELAND LTD
Ethnopharmacological relevance The stem bark of Garcinia livingstonei is used traditionally as a skin lightening agent. Aim of the study To isolate and identify compounds responsible for the observed skin lightening activity of Garcinia livingstonei and to evaluate their cytotoxicity. Materials and methods Constituents of the stem bark and fruits of Garcinia livingstonei were isolated using chromatographic techniques and structures were determined using 1D and 2D NMR and MS analysis. MeWo cells were used to evaluate the cytotoxicity and impact on melanin levels of extracts and compounds isolated, in vitro. Results Twelve known compounds, morelloflavone (1), morelloflavone-73-sulphate (2), guttiferone A (3), sargaol (4), isojacareubin (5), 6-deoxyisojacareubin (6) and in addition to the common triterpenoids, betulin, betulin aldehyde, lupeol, lupenone, euphol and stigmasterol were isolated in this investigation. Morelloflavone, morelloflavone-73-sulphate and sargaol, were found to be considerably less cytotoxic and more effective as skin lightening agents than hydroquinone. Conclusions A range of compounds was isolated from the stem bark and fruit of Garcinia livingstonei. Although the bark extract contained the cytotoxic guttiferone A, it was found to be less toxic than hydroquinone, and morelloflavone, the 73-sulphate derivative and sargaol show potential for development as depigmentation/skin lightening agents.
Koorbanally C, Crouch NR, Langlois A, Du Toit K, Mulholland DA, Drewes SE (2006) Homoisoflavanones and spirocyclic nortriterpenoids from three Eucomis species: E comosa, E schijffii and E pallidiflora subsp pole-evansii (Hyacinthaceae), SOUTH AFRICAN JOURNAL OF BOTANY 72 (3) pp. 428-433 ELSEVIER SCIENCE BV
Nawrot DA, Randrianarivelojosia M, Mulholland DA (2012) COUMARINS FROM THE MALAGASY CEDRELOPSIS RAKOTOZAFYI (PTAEROXYLACEAE), PHARMACEUTICAL BIOLOGY 50 (5) pp. 593-593
Mulholland D, Langat M, Crouch N (2012) CEMBRANOLIDES FROM CROTON GRATISSIMUS (EUPHORBIACEAE), PHARMACEUTICAL BIOLOGY 50 (5) pp. 652-652
Crouch NR, Pohl TL, Mulholland DA, Ndlovu E (2005) Alkaloids from three ethnomedicinal Haemanthus species: H. albiflos, H. deformis and H. pauculifolius (Amaryllidaceae), South African Journal of Botany 71 (1) pp. 49-52
Three closely-allied ethnomedicinal Haemanthus (Amaryllidaceae) species from the summer-rainfall region of South Africa have been phytochemically investigated and have yielded a range of isoquinoline alkaloids. H. albiflos yielded homolycorine, albomaculine and the O-methyl-lycorenium salt. From H. pauculifolius, homolycorine, the novel paucamine present as a salt, and the quaternary salts of homolycorine, montanine and manthidine were isolated. H. deformis yielded coccinine, montanine and the quaternary salt of manthidine. Copyright © NISC Pty Ltd.
Crouch NR, Langlois A, Mulholland DA (2007) Bufadienolides from the southern African Drimia depressa (Hyacinthaceae : Urgineoideae), PHYTOCHEMISTRY 68 (13) pp. 1731-1734 PERGAMON-ELSEVIER SCIENCE LTD
Koorbanally C, Sewjee S, Mulholland DA, Crouch NR, Dold A (2007) Homoisoflavanones from Pseudoprospero firmifolium of the monotypic tribe Pseudoprospereae (Hyacinthaceae : Hyacinthoideae), PHYTOCHEMISTRY 68 (22-24) pp. 2753-2756 PERGAMON-ELSEVIER SCIENCE LTD
Elgorashi EE, Coombes PH, Mulholland DA, Van Staden J (2007) Isoeugenitol, a cyclooxygenase-1 inhibitor from Gethyllis ciliaris, South African Journal of Botany 73 (1) pp. 156-158
Bioassay guided fractionation of the dichloromethane extract of the underground parts of Gethyllis ciliaris, using a cyclooxygenase-1 enzyme assay, has resulted in the isolation of the dihydroxydimethylbenzopyran-4-one isoeugenitol 1 (IC50 = 262 ¼M). The 90% methanolic extract yielded 9Z-octadec-9-enamide 3 and the novel compound isoeugenitol glycoside 2. The structures of the isolated compounds have been assigned on the basis of spectral analysis, including 1D and 2D NMR techniques. © 2006 Elsevier B.V. All rights reserved.
Salim F, Zain MM, Ahmad R, Ridzuan MSM, Langat MK, Mulholland DA (2013) Flavan-3-ols from the leaves of Malaysian Uncaria longiflora var. pteropoda (Miq.) Ridsd,Phytochemistry Letters 6 (2) pp. 236-240 Elsevier
A novel flavonoid, (-)-2R,3R-3,5,42-trihydoxyflavan-[6,7:53, 63]-23-pyranone, named uncariechin (1), was isolated from the methanolic extract of the leaves of Uncaria longiflora var. pteropoda (Miq.) Ridsd. along with the known (-)-epiafzelechin (2) and (-)-epicatechin (3), methyl 4-hydroxybenzoate and 4-hydroxybenzaldehyde, four pentacyclic oxindole alkaloids, isopteropodine, pteropodine, uncarine F and isopteropodic acid, previously found in the stems, and two coumarins, scopoletin and 3,4-dihydroxy-7-methoxycoumarin. Structures of the compounds were elucidated by 1D and 2D NMR, FTIR, UV, MS, and experimental as well as calculated electronic circular dichroism (ECD) data. Compounds 2 and 3 were evaluated for their neurotoxic and neuroprotective properties against differentiated SH-SY5Y neuroblastoma cell lines using the MTS assay. Compounds 2 and 3 did not show any neurotoxic effects but showed strong protective potential against hydrogen peroxide-induced neurotoxicity with maximum cell viability at a concentration of 1 ¼M. © 2013 Phytochemical Society of Europe.
Schwikkard SL, Waratchareeyakul W, Knirsch W, Mulholland DA (2014) Chemical constituents from Maxillaria porphyrostele (Orchidaceae), PLANTA MEDICA 80 (10) pp. 803-803 GEORG THIEME VERLAG KG
Sihra JK, Mulholland DA, Langat MK, Crouch NR, Nuzillard J-M, Thumser AE (2012) NOVEL TRITERPENOID DERIVATIVES FROM EUCOMIS BICOLOR (HYACINTHACEAE: HYACINTHOIDEAE), PHARMACEUTICAL BIOLOGY 50 (5) pp. 645-645
Elgorashi EE, Coombes PH, Mulholland DA, Van Staden J (2007) Erratum to "Isoeugenitol, a cyclooxygenase-1 inhibitor from Gethyllis ciliaris" [South African Journal of Botany 73 (2007) 156-158] (DOI:10.1016/j.sajb.2006.06.008), South African Journal of Botany 73 (2)
Waratchareeyakul W, Hellemann E, Gil R, Chantrapromma K, Langat M, Mulholland D (2017) Application of Residual Dipolar Couplings and Selective Quantitative NOE to Establish the Structures of Tetranortriterpenoids from Xylocarpus rumphii,Journal of Natural Products 80 (2) pp. 391-402 American Chemical Society
Nine triterpenoid derivatives were isolated from the heartwood of X. rumphii and were identified as xylorumphiins E (1), C (2), L (3), and M-R (4-9). Compounds 4-9 have a hemiacetal group in the triterpenoid sidechain making them impossible to purify. Purification was achieved after acetylation and subsequent separation of the epimeric mixtures of acetates, however differentiaition of the R and S epimers was not possible using standard NMR techniques. In one case, the relative configuration of a remotely located stereocenter with respect to the stereocenters in the main skeleton was unambiguously determined using residual dipolar couplings (RDCs). Dipolar couplings were collected from the sample oriented in compressed poly (methyl methacrylate) (PMMA) gels swollen in CDCl3. In another case, the relative configuration was determined using 1D selective quantitative NOE experiments. Xylorumphiin K (10), xyloccensin E, taraxer-14-en-3b-ol, (22S)-hydroxytirucalla-7,24-diene-3,23-dione and 25-hydroxy-(20S,24S)-epoxydammaran-3-one were isolated from the bark of the same plant. Compounds 3-10 are new compounds. Compounds 1-6 and xyloccensin E were tested at one concentration, 1 x 10-5 M, in the NCI59 cell one-dose screen but did not show significant activity.
Schwikkard S, Alqahtani A, Knirsch W, Wetschnig W, Jaksevicius A, Opara E, Langat M, Andriantiana J, Mulholland D (2017) Phytochemical Investigations of Three Rhodocodon (Hyacinthaceae Sensu APG II) Species,Journal of Natural Products 80 (1) pp. 30-37 American Chemical Society
The genus Rhodocodon (Hyacinthaceae sensu APG II) is endemic to Madagascar and its phytochemistry has not been described previously. The phytochemistry of three species in this genus has been investigated and eight compounds, including three bufadienolides (compounds 1, 4, and 5), a norlignan (2), and four homoisoflavonoids (compounds 3 and 6-8) have been isolated and identified. Compounds 1-3 and 6-8 have not been described previously. The COX-2 inhibitory activity of compound 6 and compound 7 acetate (compound 7A) were investigated on isolated colorectal cancer cells. Compounds 6 and 7A inhibited COX-2 by 10% and 8%, respectively, at a concentration of 12.5 mM compared to 12% for 1 mM aspirin (the positive control).
Atangana A, Toze F, Langat M, Happi E, Meva?a Mbazea L, Mulholland D, Waffo A, Sewald N, Wansi J (2017) Acridone alkaloids from Vepris verdoorniana (Excell & Mendonça) Mziray (Rutaceae),Phytochemistry Letters 19 pp. 191-195 Elsevier
Two new acridone alkaloids, verdoocridone A (1) and B (4), together with fifteen known compounds were isolated from methanol extracts of the roots and leaves of Vepris verdoorniana. The structures of all compounds were determined by comprehensive spectroscopic analyses (1D and 2D NMR, EI- and ESI? MS). The 13C NMR values of 1,2,3,5-tetramethoxy-N-methylacridone (2) and 5-methoxyaborinine (3) are also reported. The crude extracts and compounds (1-6) were tested for their antimicrobial activity. The test delivered moderate activities for crude extracts and compounds 1, 5 and 6 against the bacterium Staphylococcus aureus and the fungi Mucor meihei and Candida albicans with MIC values between 115 and 180mg/mL for extracts and between 21.3 and 29.4mM for compounds, compared to gentamycin with 0.2mM and nystatin with 5.2mM against both fungi. The determination of the radical scanvenging activity using 1,1-dephenyl-2-picrylhydrazyl (DPPH) assay gave moderate antioxidant values for all tested compounds, with IC50 between 0.29 and 0.41mM, compared to the standard 3-t-butyl-4- hydroxyanisole (BHA) displaying 0.03mM.
Aldhaher A, Langat Moses, Ndunda B, Chirchir D, Midiwo JO, Njue A, Schwikkard S, Carew M, Mulholland Dulcie (2017) Diterpenoids from the Roots of Croton dichogamus Pax,Phytochemistry 144 pp. 1-8 Elsevier
Four previously undescribed diterpenoids including two crotofolanes, crotodichogamoin A and B, and two halimanes, crothalimene A and B, a new sesquiterpenoid, and fifteen previously reported compounds, including the crotofolane, crotohaumanoxide, the casbane, depressin, a further seven furanohalimane diterpenoids, three patchoulane and two further cadinane sesquiterpenoids and aleuritolic acid were isolated from the root of Croton dichogamus. Crotodichogamoin B is an important biosynthetic intermediate of the crotofolane class and this is the first report of patchoulene sesquiterpenoids from the genus. Compounds were tested at one concentration, 1x10-5 M, in the NCI59 cell one-dose screen but did not show significant activity snd were also evaluated for their cytotoxicity against Caco-2 cell lines using the neutral red assay. 10-epi-Maninsigin D reduced Caco-2 cell viability at 10, 30 and 100 µM, with values of decreased viability of 28%, 48% and 43% respectively. None of the other tested compounds showed significant activity.
Mulholland Dulcie, Thuerig B, Langat Moses, Tamm L, Nawrot DA, James Emily, Qayyum M, Shen D, Ennis K, Jones A, Hokkanen H, Demidova N, Izotove D, Schärer H-J (2017) Efficacy of extracts from eight economically important forestry species against grapevine downy mildew (Plasmopara viticola) and identification of active constituents.,Crop Protection 102 pp. 104-109 Elsevier
Downy mildew caused by the oomycete Plasmopara viticola is the most devastating pathogen of grapevine. In this study, extracts of the bark of eight important northern forestry species were screened to find extracts showing activity against this pathogen and to identify the active compounds. Extracts from all eight species showed activity against P. viticola, the most promising, dichloromethane extracts of three Larix species, almost completely protecting grapevine from downy mildew under semi-controlled conditions. Five promising lead compounds were isolated: larixol, larixyl acetate, lariciresinol and lariciresinol acetate from Larix species and 7±,15-dihydroxydehydroabietic acid from Pinus sylvestris. These compounds showed 90%-100% efficacy. Larixol and larixyl acetate or 7±,15-dihydroxydehydroabietic acid are present in significant amounts in Larix or P. sylvestris bark extracts, respectively.
The identified active compounds are gained from a renewable resource potentially available in large quantities at relatively low prices, making them interesting candidates for the development of a plant-derived fungicide active against a key pathogen in agriculture, providing an opportunity to the forest industry to transform low value by-products into high value-added, bioactive extracts.
The Asparagaceae family is richly represented in the southern African region and has approximately 900 species. It consists of four tribes, the Hyacintheae, Urgineeae, Ornithogaleae, and Oziroeeae. Members of the Asparagaceae are widely used in traditional remedies for the treatment of several ailments, such as infections, rheumatism, inflammation and disorders associated with the central nervous system. Different species belonged to Urgineeae tribe (Rhodocodon campanulatus, R. calcicola), Hyacintheae tribe (Chionodoxa luciliae (Gigantea Hort)), Ornithogaleae tribe (Ornithogalum pyramidale and Avonsera convallarioides) have been selected for the present phytochemical study. The bulbs of R. campanulatus yielded five new compounds, a bufadienolide (RC.1), a homoisoflavonoid (RC.2) and two acetylated derivatives of a homosoflavonoid (RC.2a & RC.2b) and an acetylated neolignan (RC.3a). The bulbs of R. calcicola gave two known bufadienolides (RA.1 & RA.2). C. luciliae yielded ten compounds, including scillascillin-type homoisoflavonoids (CL.1-CL.5a), 4-chromanone (CL.4), p-hydroxybenzaldehyde (CL.6), 3-benzyl-chromanone type (CL.7-CL.9) and 3-benzylidene-4-chromanone (CL.10). O. pyramidale yielded new cardenolide glycosides (OP.1). A. convallarioides gave a known polyhydroxylated compound, polybotrin (AV.1), a purine nucleoside, adenosine (AV.2) and trans N-p-coumaroyloctopamine (AV.3). The Euphorbiaceae family is a large and diverse family which comprises 340 genera. Several species belonging to the Croton genus are employed as traditional medicinal agents in Africa, Asia and South America. The Croton species are popularly used to treat cancer, hypertension, inflammation, rheumatism, bleeding gums, malaria, asthma, diabetes and ulcers. In this study, three Croton species, C. megalocarpus, C. menyhartii and C. rivularis have been investigated for their chemistries. The roots of C. megalocarpus gave ten compounds, six diterpenoid compounds belonged to ent-clerodane (CP.1-CP.6), halimane (CP.7) and ent-abietane (CP.8-CP.9) and a triterpenoid, lupeol (CP.10). The leaves of C. menyhartii gave two flavonoids: (CM.1-CM.2) and 4-hydroxy-1-methyl-2-pyrrolidinone (CM.3) as well as large quantities of sitosterol. The leaves of C. rivularis gave one acyclic triterpene, squalene (CR.1) and an acyclic diterpenoids, phytol (CR.2). The structures of these compounds were determined using FTIR, MS, 1D, 2D NMR studies and CD analysis. Compounds RC.1 and RC.3a, a
Thueriga Barbara, James Emily, Schärer Hans-Jakob, Langat Moses, Mulholland Dulcie, Treutwein Jonas, Kleeberg Ina, Ludwig Mathias, Jayarajah Praveen, Giovannini Oscar, Markellou Emilia, Tamm Lucius (2017) Reducing copper use in the environment: The use of larixol and larixyl acetate to treat downy mildew caused by Plasmopara viticola in viticulture,Pest Management Science 74 (2) pp. 477-488 Wiley

BACKGROUND: Plant extracts might provide sustainable alternatives to copper fungicides, which are still widely used despite their unfavourable ecotoxicological profile. Larch bark extract and its constituents, larixyl acetate and larixol, have been shown to be effective against grapevine downy mildew (Plasmopara viticola) under semi-controlled conditions. The aim of this study was to reduce the gap between innovation and the registration of a marketable product, namely to develop scalable extraction processes and to evaluate and optimize performance of larch extracts under different conditions.

RESULTS: Toxicologically and technically acceptable solvents like ethanol were used to extract the active compounds larixyl acetate and larixol from bark in sufficient amounts and their combined concentration could be increased up to 39% by purification steps. The combined concentration of larixyl acetate and larixol from larch turpentine could be increased up to 66%. MIC100 against P. viticola in vitro (6-23 mg mL-1) and EC50 in planta under semi-controlled conditions (0.2-0.4 mg mL-1) were promising compared to other plant extracts. In vineyards, efficacies of larch extracts reached up to 68% in a stand-alone strategy and 84% in low-copper strategies.

CONCLUSIONS: Larch extracts represent valid candidates for copper reduction in organic vineyards, and their development into a sustainable plant protection product might be feasible

Langat Moses, Crouch Neil R., Nuzillard Jean-Marc, Mulholland Dulcie (2017) Pseudopulchellol: A unique sesquiterpene-monoterpene derived C-25 terpenoid from the leaves of Croton pseudopulchellus Pax (Euphorbiaceae),Phytochemistry Letters 23 pp. 38-40 Elsevier
A C-25 terpenoid, pseudopulchellol, a rare example of a compound formed by the combination of a monoterpenoid and a sesquiterpenoid unit, was isolated along with eudesm-4(15)-en-1²,6±-diol, its likely precursor, and nine known compounds, including six ent-kaurenoic acids, from the leaves of Croton pseudopulchellus. The Logic for Structure Determination (LSD) protocol was used to determine that pseudopelchellol consisted of the sesquiterpene, eudesm-4(15)-ene-6±-ol, attached at the C-1² position to C10 of the monoterpene, ±-pinene. The Electronic circular dichroism (ECD) studies were used to assign the absolute configuration of pseudopulchellol.
Njue Alice W., Omolo Josiah O., Cheplogoi Peter K., Langat Moses K., Mulholland Dulcie (2017) Cytotoxic Ergostane Derivatives from the Edible Mushroom Termitomyces microcarpus (Lyophyllaceae),Biochemical Systematics and Ecology 76 pp. 12-14 Elsevier
Five ergostanes, dimethylincisterol (1), 5±,8±?epidioxy-(22E,24R)-ergosta-6,9(11),22-trien-3²-ol (2), 5±,8±?epidioxy-(22E,24R)-ergosta-6,22-dien-3²-ol (3), 5±,6±-epoxy-(22E,24R)-ergosta-8(14),22-diene-3²,7±-diol (4), (22E,24R)-ergosta-7,22-diene-3²,5±,6²-triol (5) and betulinic acid (6) were isolated from the fresh fruiting bodies of the edible mushroom, Termitomyces microcarpus. The cytotoxicity of the compounds isolated was evaluated against the NCI 60 human cancer cell line panel. The degraded sterol dimethylincisterol (1) and 5±,8±?epidioxyergosta-6,22-dien-3²-ol (3) possessed cytotoxic activity at the 1 µM level.
Aldhaher A, Langat Moses, Knirsch W, Andriantiana J, Mulholland Dulcie (2018) Isoquinoline alkaloids from three Madagascan Crinum (Amaryllidaceae) species.,Biochemical Systematics and Ecology 77 pp. 7-9 Elsevier
The phytochemistry of two Madagascan Crinum species, Crinum hardyi Lehmiller and Crinum pronkii Lehmiller were investigated and compared to that of Crinum firmifolium Baker which has been investigated previously. In addition to hippadine, lycorine, hippeastrine, masonine and crinine from C. firmifolium, lycorine, crinine, and 6-hydroxycrinamine from Crinum hardyi and lycorine, 6-hydroxycrinamine and pseudolycorine from C. pronkii, the novel lycorine-relatediminium salt, the 6,7,11b,11c-didehydrolycorinium salt, was isolated from bulbs of both C. firmifolium and C. hardyi.
Benniou L, Langat Moses, Mulholland Dulcie, Benayache F, Benayache S (2018) Oleanane Triterpenoids from the Algerian Salvia phlomoides,Phytochemistry Letters 24 pp. 136-139 Elsevier
Three previously unreported triterpenoids 2±,3²,11±-trihydroxyolean-18-ene 1, 3²-acetoxy,2±,11±- dihydroxyolean-18-ene 2 and 2±-acetoxy,3²,11±-dihydroxyolean-18-ene 3 were isolated from the chloroform extract of aerial parts (leaves and flowers) of Salvia phlomoides collected from the Constantine region in the east of Algeria. Structures were determined using NMR spectroscopy and HR-ESI mass spectrometry. Compounds 1, 2 and 3 were tested against the NCI panel of human tumour cell lines at a single dose of 10 µM. Compound 2 was the most active showing a 38 % growth inhibition against the leukemia RPMI-8226 line.
Schwikkard S, Whitmore Hannah, Corson T, Sishtla K, Langat Moses, Carew M, Mulholland Dulcie (2018) Antiangiogenic Activity and Cytotoxicity of Triterpenoids and Homoisoflavonoids from Massonia pustulata and Massonia bifolia,Planta Medica 84 (09/10) pp. 638-644 Georg Thieme Verlag
The Hyacinthaceae family (sensu APGII) with approximately 900 species in around 70 genera, plays a significant role in traditional medicine in Africa as well as across Europe and the Middle and Far East. The dichloromethane extract of the bulbs of Massonia pustulata (Hyacinthaceae sensu APGII) yielded two known homoisoflavonoids, (R)-5-hydroxy-3-(4-hydroxybenzyl)-7-methoxy-4-chromanone 1 and 5-hydroxy-3-(4-hydroxybenzyl)-7-methoxy-4-chromone 2 and four spirocyclic nortriterpenoids, eucosterol 3, 28-hydroxyeucosterol 4 and two previously unreported triterpenoid derivatives, (17S,23S)-17±,23-epoxy-3²,22²,29-trihydroxylanost-8-en-27,23-olide 5 and (17S, 23S)-17±,23-epoxy-28,29-dihydroxylanost-8-en-3-on-27,23-olide 6. Compounds 1, 2, 3, and 5 were assessed for cytotoxicity against CaCo-2 cells using a neutral red uptake assay. Compounds 1, 2 and 5 reduced cell viability by 70% at concentrations of 30, 100 and 100 ¼M respectively. Massonia bifolia yielded three known homoisoflavonoids, (R)-(4?-hydroxy)-5-hydroxy-7-methoxy-4-chromanone 1, (R)-(4?-hydroxy)-5,7-dihydroxy-4-chromanone 7 and (R)-(3?-hydroxy-4?-methoxy)-5,7-dihydroxy-4-chromanone 9, two previously unreported homoisoflavonoids, (E)-3-benzylidene-(3?,4?-dihydroxy)-5-hydroxy-7-methoxy-4-chromanone 8 and (R)-(3?,4?-dihydroxy)-5-hydroxy-7-methoxy-4-chromanone 10, and a spirocyclic nortriterpenoid, 15-deoxoeucosterol 11. Compounds 1, 1Ac, 7, 8, 9 and 10 were screened for antiangiogenic activity against human retinal microvascular endothelial cells. Some compounds showed dose-dependent antiproliferative activity and blocked endothelial tube formation, suggestive of antiangiogenic activity.
Langat Moses, Helfenstein A, Horner C, Tammel P, Hokkanen H, Izotov D, Mulholland Dulcie (2018) Pumilol, a Diterpenoid with a Rare Strobane Skeleton from Pinus pumila (Pinaceae),Chemistry and Biodiversity 15 (10) e1800056 Wiley
Pumilol (1), a strobane diterpenoid, reported herein for the first time was isolated from the bark of Pinus pumila (Pall.) Regel (Siberian Dwarf Pine or Japanese Stone Pine), along with seventeen known compounds including serratane triterpenoids, not previously reported from this species, and four ferulate derivatives. The stereostructure of pumilol was established using HRESIMS, NMR, the DP4+ probabilities and by comparison of the experimental and calculated electronic circular dichroim (ECD) spectra. Labda-8(17),13-dien-15-oic acid (4), bornyl trans-4-hydroxycinnamate (14) and E-bornyl ferulate (15) showed activity against S. aureus and/or E. faecalis with MIC90 values 12.5-50 µM.
Control of pathogens by means of plant-derived plant protection products (PPPs) can be an effective, sustainable and environmentally friendly method of crop protection in organic agriculture. Larix decidua (European larch) bark is a by-product of the wood processing industry and provides a readily available, low cost and sustainable raw material. It was therefore planned to commercialise Larix extracts, containing two highly active diterpenoids (larixyl acetate and larixol) into a marketable PPP, Larixyne®, to combat grapevine downy mildew (Plasmopara viticola) in organic farming.

This thesis outlines the processes and knowledge developed in order to facilitate the commercialisation of Larixyne®:
(i) Analytical protocols using high-performance liquid chromatography (HPLC) and gas chromatography (GC-FID) techniques to quantify larixyl acetate, larixol and epimanool (the primary impurity) in Larix extracts were developed.
(ii) Methods for large scale extraction and purification of active compounds from Larix were optimised in line with EU specifications for products with organic agricultural use. Through extraction with 2-amino-2-methyl-1-propanol (AMP), Larix bark contained extract compositions of up to 30.53% larixyl acetate and 8.45% larixol. Larixyl acetate and larixol extracts were isolated in sufficient quantity (8.0 kg) for large scale field trials.
(iii) High-yield sources of larixyl acetate and larixol were evaluated and identified through screening a range of plant samples from central and northern Europe. Larix samples were found to vary in content of larixyl acetate (median 0.017% w/w, range 0.000 ? 4.544% w/w), larixol (median 0.003% w/w, range 0.000 ? 0.578% w/w), and epimanool (median 0.009% w/w, range 0.000 ?0.752% w/w).
(iv) The activity of larixyl acetate, larixol, epimanool and formulated products of Larix extracts against P. viticola on Vitis vinifera were demonstrated. Larixyl acetate and larixol exhibited excellent activity in vitro (mean MIC100 of 7 and 16 ¼g/mL) and in planta (EC50 0.2 ? 0.7 mg/mL).

In this work two lichen species (Cladonia portentosa and Ramalina farinacea) were investigated. Lichen have been used as antimicrobials, perfume and dye ingredients. Five known compounds were isolated from C. portentosa and seven known compounds were isolated from R. farinacea. Antibacterial activity investigation was carried out for four compounds isolated from these lichen the compounds were: (-)-usnic acid (43), olivetocarboxylic acid (45), perlatolic acid (46) and atranorin (49). Their activity was investigated against seven different bacteria, Escherichia coli (90, 91, 93, 94, 98), Salmonella Enteritidis (446, 496), Klebsiella pneumonia (97), Acinetobacter baumannii (210, 211), Pseudomonas aeruginosa (2, 92), Staphylococcus aureus (NCTC 12493) and Enterococcus faecalis (ATCC 29212). All compounds were active at high concentrations.
In Larix decidua bark two active compounds (larixol and larixyl acetate) have previously been isolated and are being developed for use in the agrochemical Larixyne ®. To extract the compounds traditional solvents such as methanol were used. In this work, two environmentally friendly natural deep eutectic solvents, choline chloride / lactic acid (1:1 mol) and choline chloride / malonic acid (1:1 mol), were investigated. It was determined that the NADES successfully extract larixol and larixyl acetate from ground L. decidua bark.
Antimicrobial resistance is a global threat, with some pathogens demonstrating resistance to all currently available commercial antimicrobials. With few novel antimicrobials in the pharmaceutical pipeline, treatments for bacterial infections, particularly those caused by Gram-negative bacteria, are extremely limited. In this work synergy between the natural product vanillin and the commercially available antibiotic colistin was investigated against various Gram-negative bacteria (Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Salmonella Enteritidis, Salmonella Typhimurium and Esherichia coli). It was determined that the combination has a synergistic
relationship against Gram-negative bacteria. Traditionally, the leaves of Sambucus ebulus are used to treat Newcastle?s Disease Virus in chicken. In this work the antiviral activity of the oil of the S. ebulus plant, its main component (isovaleric acid) and three compounds of a similar structure to isovaleric acid (menthyl isovalerate, benzyl isovalerate and levulinic acid) were investigated. Three main types of fluorometric antiviral assays were carried out: combined, therapeutic and prophylactic. The oil was found to be active against NDV therefore supporting its traditional use. Isovaleric acid and other compounds were found to have some activity against NDV.
Aziz Ahmad Nazif, Ismail Nor Hadiani, Abdul Halim Siti Nadiah, Loo Chung Yeng, El Hassane Anouar, Langat Moses K, Mulholland Dulcie, Awang Khalijah (2018) Laevifins A-G, Clerodane Diterpenoids from the Bark of Croton oblongus Burm.f.,Phytochemistry 156 pp. 193-200 Elsevier
A phytochemical investigation of the stem barks of the Malaysian Croton oblongus Burm.f. (Syn. Croton laevifolius Blume) (Euphorbiaceae) yielded seven previously undescribed ent-neo-clerodane diterpenoids, laevifins A - G (1, 2, 4-8) and the known crovatin (3). Structures were established by a combination of spectroscopic methods including HRESIMS, NMR spectroscopy and X-ray crystallography. The absolute configuration of crovatin and laevifins A-G was established by comparison of experimental ECD and theoretical TDDFT ECD calculated spectra. This is the first report on the occurrence of the sesquiterpenoid cryptomeridiol in a Croton species. In vitro cytotoxicity assays on laevifins A (1), B (2) and G (8) showed moderate activities against the MCF-7 cancer cell line (IC50 102, 115 and 106 ¼M, respectively) while ²-amyrin and acetyl aleuritolic acid showed good anti-inflammatory activity on the LPS-induced NF-ºB translocation inhibition in RAW 264.7 cells assay with IC50 values of 23.5 and 35.4 ¼g/mL, respectively.
Pathogenic neovascularization underlies a number of degenerative ocular diseases, which together are the most common causes of blindness in children and adults. Novel therapies are sorely needed to treat these diseases as even the newer, highly expensive, anti-vascular endothelial growth factor (VEGF) drugs, such as ranibizumab, can have acute, systemic side effects.
Natural product chemistry involves the extraction of potentially pharmacologically active phytochemicals from plants. Homoisoflavonoids are a class of active compounds isolated from the Scilloideae sub-family of the Asparagaceae family. Homoisoflavonoids are frequently extracted from the Scilloideae sub-family and are considered promising pharmacological candidates due to their anti-bacterial, anti-mutagenic, anti-oxidant and anti-inflammatory effects. Cremastranone is a homoisoflavonoid extracted from an orchid used in traditional medicine, Cremastra appendiculata, and has an anti-proliferative effect on blood vessels. This compound has been shown to be effective in treating pathogenic neovascularisation in premature mice. A synthetic method, which can be used to produce a wide variety of homoisoflavonoid compounds, is reported here with the goal to synthesise the most promising homoisoflavonoid candidates extracted in this work for in vivo studies.
The sub-family Scilloideae contains the tribes Ornithogaleae, Hyacintheae, Urgineeae and Oziroëeae. Seven plant species from the Hyacintheae, Urgineeae and Oziroëeae tribes were investigated in this work. This is the first report of the phytochemical investigation of Rhodocodon cryptopodus, R. rotundus, Massonia (Whiteheadia) bifolia and Oziroë arida.
Rhodocodon cryptopodus (Baker) and Rhodocodon rotundus (H. Perrier) bulbs were investigated, producing six known compounds identified as triterpenoids, cinnamic acid derivatives and a homoisoflavonoid and four novel compounds. From R. cryptopodus one novel coumarin was extracted and from R. rotundus, two novel bufadienolides and one novel 3S-homoisoflavonoid were extracted
Plants within the Eucomis (L'Her) genus are known to contain many homoisoflavonoids. E. autumnalis (Mill. Chitt.), E. bicolor (Baker) and E. comosa (Hort ex. Wehrh) were investigated. These species have been investigated previously. They were reinvestigated using bulbs from a commercial bulb supplier as large quantities were available and large amounts of extracted compounds were required for screening purposes. Phytochemical investigations yielded a spirocyclic nortriterpenoid, phenolic compounds, a chromanone, a purine nucleoside and homoisoflavonoids. One novel homoisoflavonoid was extracted from E. autumnalis and two novel homoisoflavonoids were extracted from E. comosa.
The phytochemistry of Massonia (Whiteheadia) bifolia ((Jacq.) J.C. Manning & Goldblatt) was investigated. Analysis of the bulbs of M. bifolia yielded four known compounds including a spirocyclic nortriterpenoid and three homoisoflavonoids. Novel compounds included a xanthone and two homoisoflavonoid compounds.
A phytochemical analysis of Oziroë arida (Raf.) bulbs yielded six known compounds, identified as two phytosterols, a cinnamic acid derivative, a purine nucleoside and two homoisoflavonoids.
Extracted and synthetic homoisoflavonoids were tested for their anti-proliferative activities in cell-based assays. Active compounds were also tested for their anti-angiogenic activities in vitro. Results indicated that several homoisoflavonoids are very active at stopping the proliferation, migration and tube formation of cultured endothelial cells. Selected promising compounds will be taken on for in vivo studies in the future.
This work considers interactions occurring between humic acid (HA) and other representative components of oil sands such as sand and clays, and their potential effect on bitumen recovery. Further, it considers the characterisation of oil sand components in terms of physico-chemical and dielectric properties. The study focuses on a combination of artificially prepared systems analysed using a variety of techniques, including dielectric spectroscopy (DS), UV-Vis spectrophotometry (UV-Vis), sedimentation analysis and adsorption isotherms. Finally, it talks about HA extraction from natural oil sands, HA behaviour in solution and its interactions with oil sand components.

Dielectric properties arise from electronic and atomic polarisation or dipole, ionic or dielectric relaxation mechanisms. The particular sensitivity of basic electrical measurements, e.g. resistivity, to the presence of water made the original use of this approach most relevant to the determination of water-filled porosity in model oil sand samples. The present study builds on recent developments in the literature, and, specifically determines the extent to which the potentially dominant effects of water can be overcome in order to access additional information, such as wettability. DS was used to study physico-chemical and structural characteristics of artificially prepared simplified oil sand systems. At the low-frequency DS analysis (10-3 ? 107 Hz) yields complex properties, which revealed contributions from underlying processes in materials, including electronic, ionic and interfacial polarisation. Dielectric measurements undertaken on a range of artificially prepared samples revealed strong, regular dependencies on the composition of the systems. This model study proved DS to be a valid method for the analysis of complex systems such as synthetically prepared oil sands. The results indicated wettability change in a systematic manner in samples made up of varying combinations of sand, clays, water and bitumen.

It has been considered that water-soluble organic species can influence the recovery of bitumen by having an effect on parameters such as wettability or surface charge, thereby altering interactions between oil sand components. HA is of considerable environmental significance, being a major component of soil. However, its structure and colloidal properties continue to be the subject of debate, largely owing to its molecular complexity and association with other humic substances and mineral matter. Natural oil sand (NOS) samples obtained from Alberta, Canada were separated into solid, oil and water fractions. HA was further extracted out of water fractions using a range of procedures. The obtained organic species were identified mainly using Fourier Transform infrared (FTIR) spectroscopy and compared against standard components. The presence of aromatics, alcohols, phenols, ketones and aliphatic and aromatic amines was confirmed in the extracted HA samples.

An adsorption study of HA on mineral surfaces was done by means of conductivity measurements. The adsorption of various concentrations of NaHA from the aqueous solutions onto the surface of sand, clay and sand-clay mixtures with varying ratios was studied by means of DS and the UV-vis absorption measurements. The results showed a strong affinity of the HA present in the solutions to the surface of sand and clay. The calculations based on the results obtained proved the significance of clay in the HA-solids interactions. Although the sand surface covered in HA remains naturally hydrophilic (slightly less than a natural sand), it hinders the adsorption of asphaltenes onto the surface. The HA adsorbs onto the asphaltenes coated sand, making the surface increasingly more hydrophobic over time.

Colloid and interfacial properties were studied by means of conductivity, surface tension and zeta potential measurements. a class, HA is considered to comprise supramolecular assemblies of heterogeneous spec

Tabekoueng Georges Bellier, Akak Carine Mvot, Langat Moses K., Blaise Azebaze Anatole Guy, Kamdem Waffo Alain Francois, Mulholland Dulcie A., Vardamides Juliette Catherine (2019) The chemical constituents of Penianthus longifolius Miers,Phytochemistry Letters 30 pp. 103-106 Elsevier
Fourteen compounds were isolated from the roots, leaves and twigs of Penianthus longifolius Miers (Menispermaceae), including two previously unreported neo-clerodane diterpenoids, penianthin (1), its C-8 epimer (2). In addition, the previously reported O-methylmoschatoline (3), taraxerol (4), taraxerone (5), rubrosterone, (6), panuosterone (7), 22-epi-20-hydroxyecdysone (8), ergosterol peroxide (9), stigmast-5-ene-3,7-dione (10), stigmasterol (11), ²-sitosterol (12), stigmasterol glucoside (13) and ²-sitosterol glucoside (14) were isolated. The structures of the compounds were determined by means of NMR spectroscopic and mass spectrometric analysis. The absolute configurations of 1 and 2 were determined by circular dichroism analysis. Compounds 1 and 2 were screened against the NCI60 cancer cell panel but showed no significant activity at 10 ¼M.
Kalantarzadeh Mina, Mulholland Dulcie, De Leij Frans, Webber Joan F. (2019) Induced antimicrobial activity in heat-treated woodchips inhibits the activity of the invasive plant pathogen Phytophthora ramorum,Plant Pathology 68 (5) pp. 889-900 Wiley
We investigated the antimicrobial activity of heat?treated woodchips of three woody host species against the invasive oomycete plant pathogen Phytophthora ramorum to assess the potential of heated woodchips for disease suppression. Results demonstrated that heat?treated pine (Pinus sylvestris), Japanese larch (Larix kaempferi) and rhododendron (Rhododendron ponticum) woodchips inhibited the recovery of P. ramorum spores and mycelium compared with similar material that had only been air?dried. Effects were most evident with pine and larch; inhibition was maintained even when larch woodchips were diluted with soil. In vitro assays using methanol crude extracts from woodchips of the three species showed they all had an inhibitory effect on P. ramorum zoospores and reduced chlamydospore germination compared with air?dried wood extracts. Chemical analysis of the extracts revealed several induced compounds were present but in different concentrations for each species. Coniferaldehyde was the most active inhibitory against spores and mycelium, whilst the dominant resin acids, dehydroabietic and abietic acid, decreased the minimum inhibitory concentration of phenolic compounds tested against P. ramorum but were ineffective when used alone. An array of compounds, including dehydroabietic acid, methyl abietate, ±?pinene and 3?carene, occurred at elevated levels in the living tissue of Japanese larch bark attacked by P. ramorum. These compounds may be part of the induced resistance response of larch to P. ramorum. Results of a field trial using heat?treated and air?dried woodchips were consistent with the crude extract bioassay results, suggesting that heat?treated woody materials have potential to reduce the survival of P. ramorum under natural conditions.
Schwikkard Sianne, Whitmore Hannah, Sishtla Kamakshi, Sulaiman Rania S., Shetty Trupti, Basavarajappa Halesha D., Waller Catherine, Alqahtani Alaa, Frankemoelle Lennart, Chapman Andy, Crouch Neil, Wetschnig Wolfgang, Knirsch Walter, Andriantiana Jacky, Mas-Claret Eduard, Langat Moses K., Mulholland Dulcie, Corson Timothy W. (2019) The Antiangiogenic Activity of Naturally Occurring and Synthetic Homoisoflavonoids from the Hyacinthaceae (sensu APGII),Journal of Natural Products American Chemical Society
Excessive blood vessel formation in the eye is implicated in wet age-related macular degeneration, proliferative diabetic retinopathy, neovascular glaucoma, and retinopathy of prematurity, which are major causes of blindness. Small molecule antiangiogenic drugs are strongly needed to supplement existing biologics. Homoisoflavonoids have been previously shown to have potent antiproliferative activities in endothelial cells over other cell types. Moreover, they demonstrated a strong antiangiogenic potential in vitro and in vivo in animal models of ocular neovascularization. Here, we tested the antiangiogenic activity of a group of naturally occurring homoisoflavonoids isolated from the family Hyacinthaceae and related synthetic compounds, chosen for synthesis based on structure?activity relationship observations. Several compounds showed interesting antiproliferative and antiangiogenic activities in vitro on retinal microvascular endothelial cells, a disease-relevant cell type, with the synthetic chromane, 46, showing the best activity (GI50 of 2.3 × 10?4 ¼M).
Three new homoisoflavonoids, (3R)-3,5-dihydroxy-7-methoxy-3-(4?-hydroxybenzyl)-4-chromanone, 4, and its 5-O-²-D-glucopyranoside and 5-O-²-D-gentiobiose derivatives, 5 and 6, along with the known homoisoflavonoids, (3S)-5-hydroxy-7-methoxy-3-(4-methoxybenzyl)-4-chromanone, 1, (3R)-5-hydroxy-7-methoxy-3-(4-methoxybenzyl)-4-chromanone (2) and (3S)-5-hydroxy-7-methoxy-3-(4-hydroxybenzyl)-4-chromanone, 3, were isolated from the bulbs of Ornithogalum dubium Houtt.. In addition, three known cardenolides, 3²-(O-±-L-rhamnoside)-5²,14²-dihydroxy-19-oxocardenolide 7, 3²-(O-±-L-rhamnoside)-5²,11±,14²-trihydroxy-19-oxocardenolide, 8 and 3²-(O-±-L-rhamnoside)-5²,11±,14²,19-tetrahydroxycard-20(22)-enolide, 9, were isolated from Ornithogalum ponticum ?Sochi?. Yields of homoisoflavonoids were low (0.0002 % - 0.0003 %) so stereospecific synthetic methodology needs further refinement to produce antiangiogenic (3R)-3-benzylchromanones and (3S)-3-hydroxyhomoisoflavonoid analogues for further evaluation. This work confirms that, apart from producing cardenolides, the absolute configuration at C-3 of homoisoflavonoids produced by Ornithogalum is 3R (H-3²) for the 3-benzylchromanones and 3S (3-OH ²) for the 3-hydroxy benzylchromanones, in agreement with our earlier reports.
Isyaka Sani M., Langat Moses K., Mas-Clare Eduard, Mbala Blaise M., Mvingu Bienvenu K., Mulholland Dulcie A. (2019) Ent-abietane and ent-pimarane diterpenoids from Croton mubango (Euphorbiaceae),Phytochemistry 170 112217 Elsevier
Twelve ent-abietane and two ent-pimarane diterpenoids were isolated from the leaves of Croton mubango Müll. Arg. (Euphorbiaceae) collected in the Democratic Republic of the Congo. 2²-Hydroxy-ent-abieta-7,13-dien-3-one, 15-hydroxy-ent-abieta-7,13-dien-3-one, 13±,15-dihydroxy-ent-abieta-8(14)-en-3-one, 2²,9,13-trihydroxy-ent-abieta-7-en-3-one, 2²,7²-dihydroxy-ent-abieta-8,11,13-trien-3-one, 15-hydroxy-ent-abieta-8,11,13-trien-3-one and ent-pimara-8(14),15-dien-3-one and the ent-forms of the previously reported normal series diterpenoids, ent-abieta-8,11,13-trien-3-one, 7²-hydroxy-ent-abieta-8,11,13-trien-3-one, 3±-hydroxy-ent-abieta-8,11,13-triene, 15-hydroxy-ent-abieta-8,11,13-triene and 6²-hydroxy-ent-abieta-8,11,13-triene are reported here for the first time. Structures were established using HRESIMS, FTIR, NMR, DP4+ probability calculations and by comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Ent-pimara-8(14), 15-dien-3-one, showed antiproliferative activity against melanoma (MALME-3M), renal (UO-31) and ovarian cancer cell lines (IGROV1) at a concentration of 10?5 M in the NCI 60 screen.
The genus Croton consists of an important group of trees, with several of its species being commercially exploited either as medicines, biofuels or for the cosmetics industry. Plaunotol, an anti-peptic ulcer drug marketed as Kelnac® is isolated from the leaves of Croton stellatopilosus, Sangre De Drago from Croton lechleri is commercially available for the treatment of wounds and diarrhoea, and Croton megalocarpus seed oil is commercially available as a biofuel and for cosmetic uses. Several species of the Croton genus have been reported as being used ethnomedicinally to treat hypertension, rheumatism, inflammation, bleeding gums, malaria, diarrhoea, syphilitic ulcers, asthma, pain and diabetes. The genus Croton is one of the largest genera of the Euphorbiaceae sensu stricto, and consists of over 1300 species of shrubs and trees that are distributed in the warm tropics and sub-tropics. In Africa, there are more than 292 Croton species where 124 are reported to occur in continental Africa, 156 in Madagascar and 12 in the Indian Ocean islands of Reunion, Comoros, Mauritius and Sao Tome and Principe.
This study focussed on the chemistry and pharmacological activity of three Central and East African Croton species, Croton mubango Mull. Arg. and Croton haumanianus J. Leonard from the Democratic Republic of Congo and Croton dictyophlebodes Radcl.-Sm from Tanzania. These species are traditionally used in the treatment of a variety of diseases. A total of seventy-five compounds were isolated from these species, including sixty diterpenoids, three triterpenoids, three sesquiterpenoids, three phytosterols, two phenolic compounds, one ferulic acid derivative, linoleic acid and two chlorophyll derivatives. The chemical structures of the isolated compounds were determined using NMR spectroscopy, mass spectrometry, and infrared and specific rotation experiments. DP4+ calculations were used to determine the absolute configuration of compound CM-6 whereas TDDFT-ECD simulation experiments were used to determine absolute configurations for compounds CM-1, CM-2, CM-5, CM-17, CH-19 and CH-21. Thirty-eight diterpenoids are described in this study for the first time, including eleven ent-abietane and one ent-pimarane diterpenoids from C. mubango, nine ent-kauranes, three ent-clerodanes and five ent-isopimaranes from C. haumanianus, and eight ent-clerodanes and one ent-trachylobane from C. dictyophlebodes. Thirty diterpenoids were submitted to the Development Therapeutics Program (DTP) of the National Cancer Institute (NCI) in the United States of America for the NCI 60 anticancer cell line screening programme. Compounds CM-17, CH-11, CH-12, CH-13 and CH-15 showed selective antiproliferative effects against three of the NCI 60 cancer cell lines. Compounds CH-11, CH-12, CH-13 and CH-15 were found to show 100% lethality against colon (HCT-116), melanoma (M14) and renal (786-0) cancer cell lines whereas CM-17 gave 99%, 89% and 82% cell lethality against melanoma (MALME-3M), renal (UO-31) and ovarian cancer (IGROV1) cell lines respectively at a concentration of 10-5 M.
Langat Moses K., Dlova Ncoza C., Mulcahy-Ryan Lauren E., Schwikkard Sianne L., Opara Elizabeth I., Crouch Neil R., Hiles Jacob D., Mulholland Dulcie (2020) The effect of isolates from Cassipourea flanaganii (Schinz) Alston, a plant used as a skin lightening agent, on melanin production and tyrosinase inhibition,Journal of Ethnopharmacology 113272 Elsevier
Ethnopharmacological relevance: The Zulu and Xhosa people of South Africa use the stem bark of Cassipourea flanaganii as a skin-lightening cosmetic.
Aim of the study: To isolate and identify compounds responsible for the skin lightening properties from the stem bark of Cassipourea flanaganii and to evaluate their cytotoxicity towards skin cells, and potential to treat hyperpigmentation.
Materials and methods: Extracts from the stem bark of Cassipourea flanaganii were isolated using chromatographic methods and structures were determined using NMR, IR and MS analysis. The tyrosinase inhibitory activity and the ability to inhibit the production of melanin were determined using human primary epidermal melanocyte cells. Cytoxicity was established using the same melanocytes and a neutral red assay.
Results: One previously undescribed compound, ent-atis-16-en-19-al (1) along with the known ent-atis-16-en-19-oic acid (2), ent-atis-16-en-19-ol (3), ent-kaur-16-en-19-oic acid (4), ent-kaur-16-en-19-al (5), ent-manoyl oxide (6), guinesine A (7), guinesine B (8), guinesine C (9), lichenxanthone (10), 2,4-dihydroxy-3,6-dimethyl benzoic acid methyl ester (11), lynoside (12), lupeol (13), ²-amyrin (14), docosyl ferulate (15), stigmasterol, sitosterol and sitosterol-O-glucoside were isolated in this investigation. An impure fraction containing compound 3 was acetylated to obtain 19-acetoxy-ent-atis-16-ene (3a). Compounds 10 and 11 are usually isolated from lichen, hence they are possible contaminants of lichen harvested with the bark. Compounds 1, 3a, 5-14 were not significantly cytotoxic to the primary epidermal melanocyte cells (P > 0.05) when compared to the negative and positive controls (DMSO, 0.1% and hydrogen peroxide, 30 wt% in water). Inhibition of tyrosinase was significantly greater with respect to the negative control (P Conclusions: Compounds 1, 5-14, isolated from Cassipourea flanaganii and the derivative 3a showed low cytotoxicity. All compounds had a clear time and concentration dependent effect on melanin content which did not appear to be dependent on their inhibition of tyrosinase.
Takou Dieudonné M., Waffo Alain F. Kamdem, Langat Moses K., Wansi Jean D., Mulcahy-Ryan Lauren E., Schwikkard Sianne L., Opara Elizabeth I., Mas-Claret Eduard, Mulholland Dulcie (2019) Melanin Production Inhibitors from the West African Cassipourea congoensis,Planta Medica International Open 6 (02) pp. e50-e56 Thieme Open
Cassipourea congoensis (syn. Cassipourea malosana) is used in African countries as a skin-lightening agent. Two previously unreported cycloartane triterpenoids, 26-hydroxy-3-keto-24-methy lenecycloartan-30-oic acid 1 and 24-methylene-cycloartan-3²,26,30-triol 2 along with the known mahuannin B 3, 7-methoxymahuannin B 4, 7-methoxygeranin A 5, methyl-3-(4-hydroxy-3-methoxyphenyl)-2E-propenoate, glycerol-1-alkanoate, (E)-3-(4-hydroxy-3-methoxyphenyl)prop-2-enal 6, (-)-syringaresinol 7, and stigmast-5-en-3-O-²-D-glucoside, were isolated from the roots of C. congoensis. The crude extract and compounds 1 and 5 were found to inhibit the production of melanin at 10 ¼M with low cytotoxicity validating the ethno-medicinal use of this plant
Munissi Joan J.E., Isyaka Sani M., Mas-Claret Eduard, Brabner Molly, Langat Moses K., Nyandoro Stephen S., Mulholland Dulcie A. (2020) Ent-clerodane and ent-trachylobane diterpenoids from Croton dictyophlebodes,Phytochemistry 179 112487 Elsevier
The stem bark and root bark extracts of Croton dictyophlebodes (Euphorbiaceae) yielded seven undescribed ent-clerodanes: 15,16-epoxy-17,12(S)-olide-ent-cleroda-1,3,13(16),14-tetraen-18-oic acid methyl ester (crotodictyo A), 3²,4²:15,16-diepoxy-ent-cleroda-13(16),14-dien-20-al (crotodictyo B), 3²,4²:15,16-diepoxy-ent-cleroda-13(16),14-dien-19,20-dioic acid (crotodictyo C), 3²,4²:15,16-diepoxy-ent-cleroda-13(16),14-dien-20,19-olide (crotodictyo D), 3²,4²:15,16-diepoxy-20,12(R)-olide ent-cleroda-13(16),14-dien-19-oic acid methyl ester (crotodictyo E), 15,16-epoxy-ent-cleroda-3,13(16),14-trien-12-oxo-18-oic acid (crotodictyo F) and 15,16-epoxy-ent-cleroda-1,3,13(16),14-tetraen-12-oxo-18-oic acid (crotodictyo G), in addition to 15,16-epoxy- ent-cleroda-3,13(16),14-trien-12-oxo-18-oic acid methyl ester (crotodictyo H), reported previously as a synthetic derivative, and acetyl aleuritolic acid. The root extract yielded two ent-trachylobanes, ent-trachylobane-18,19-diol, the undescribed ent-trachylobane-2±,19-diol, along with ent-kaur-16-en-19-oic acid and 2-methoxybenzyl benzoate. Compounds were evaluated against the NCI 60 panel of human tumour cell lines at a single dose of 10?5 M, but showed no significant activity.
Isyaka Sani M., Mas-Claret Eduard, Langat Moses K., Hodges Thomas, Selway Bethany, Mbala Blaise M., Mvingu Bienvenu K., Mulholland Dulcie (2020) Cytotoxic diterpenoids from the leaves and stem bark of Croton haumanianus (Euphorbiaceae),Phytochemistry 178 112455 Elsevier
The leaf extract of Croton haumanianus J. Léonard (Euphorbiaceae) yielded twenty-six compounds, including eight previously reported ent-kauranes and an ent-labdane and eight undescribed ent-kauranes, ent-16R-kauran-17-al, ent-3²-hydroxy-16R-kauran-17-al, ent-16S,17-epoxykauran-19-ol, ent-16S,17-epoxykauran-3²-ol, ent-17-palmityloxykaurane-3²,16²-diol, ent-17-palmityloxykauran-16²-ol, ent-3±,18-cyclokaurane-16²,17-diol and 19-nor-16±,17-dihydroxy-ent-kaur-4(18)-ene and three undescribed ent-clerodanes, dimethyl ent-15,16-epoxy-6²-hydroxy-1,3,13(16),14-clerodatetraen-20,12S-olide-18,19-dioate (saniolide A), dimethyl ent-15,16-epoxy-6²-hydroxy-1,3,13(16),14-clerodatetraen-20,12R-olide-18,19-dioate (12-epi-saniolide A), methyl ent-15,16-epoxy-1,3,13(16),14-clerodatetraen-18,6R:20,12S-diolide-19-oate (saniolide B) . The stem bark extract yielded the ent-clerodane crotocorylifuran, and five undescribed ent-isopimaranesent-isopimara-8(14),15-dien-18-al, ent-18-hydroxyisopimara-8(14),15-dien-7-one, ent-isopimara-7,15-dien-18-oic acid, ent-isopimara-7,15-dien-18-ol and ent-isopimara-8,15-dien-7-oxo-18-oic acid. Three compounds, ent-kaurane-3²,16²,17-triol, ent-17-palmityloxykaurane-3²,16²-diol and ent-17-palmityloxykauran-16²-ol, showed selective activity against three of the NCI 60 cancer cell lines, the colon (HCT-116), the melanoma (M14) and the renal (786-0) cancer cell lines at a concentration of 10-5 M.
Whitmore Hannah, Sishtla Kamakshi, Knirsch Walter, Andriantiana Jacky L., Schwikkard Sianne, Mas-Claret Eduard, Nassief Sarah M., Isyaka Sani M., Corson Timothy W., Mulholland Dulcie A. (2020) Bufadienolides and anti-angiogenic homoisoflavonoids from Rhodocodon cryptopodus, Rhodocodon rotundus and Rhodocodon cyathiformis,Fitoterapia 141 104479 Elsevier

Background

Homoisoflavonoids have been shown to have potent anti-proliferative activities in endothelial cells over other cell types and have demonstrated a strong antiangiogenic potential in vitro and in vivo in animal models of ocular neovascularization. Three species of Rhodocodon (Scilloideaea subfamily of the Asparagaceae family), endemic to Madagascar, R. cryptopodus, R. rotundus and R. cyathiformis, were investigated.

Purpose

To isolate and test homoisoflavonoids for their antiangiogenic activity against human retinal microvascular endothelial cells (HRECs), as well as specificity against other ocular cell lines.

Methods

Plant material was extracted at room temperature with EtOH. Compounds were isolated using flash column chromatography and were identified using NMR and CD spectroscopy and HRESIMS. Compounds were tested for antiproliferative effects on primary human microvascular retinal endothelial cells (HRECs), ARPE19 retinal pigment epithelial cells, 92?1 uveal melanoma cells, and Y79 retinoblastoma cells. HRECs exposed to compounds were also tested for migration and tube formation ability.

Results

Two homoisoflavonoids, 3S-5,7-dihydroxy-(32-hydroxy-42-methoxybenzyl)-4-chromanone (1) and 3S-5,7-dihydroxy-(42-hydroxy-32-methoxybenzyl)-4-chromanone (2), were isolated along with four bufadienolides. Compound 1 was found to be non-specifically antiproliferative, with GI50 values ranging from 0.21?0.85 ¼M across the four cell types, while compound 2 showed at least 100-fold specificity for HRECs over the other tested cell lines. Compound 1, with a 3S configuration, was 700 times more potent that the corresponding 3R enantiomer recently isolated from a Massonia species.

Conclusion

Select homoisoflavonoids have promise as antiangiogenic agents that are not generally cytotoxic.

Waratchareeyakul Watcharee, Fusi Fabio, Durante Miriam, Ahmed Amer, Knirsch Walter, Mas-Claret Eduard, Mulholland Dulcie A. (2020) Vasorelaxing Activity of Stilbenoid and Phenanthrene Derivatives from Brasiliorchis porphyrostele: Involvement of Smooth Muscle CaV1.2 Channels,Planta Medica 86 (09) pp. 631-642 Thieme Gruppe
Five compounds, 3,4?-dihydroxy-3?,5,5?-trimethoxydihydrostilbene, 1, 3,4'-dihydroxy-3',5'-dimethoxydihydrostilbene, 2, 3,4'-dihydroxy-5,5'-dimethoxydihydrostilbene, 3, 9,10-dihydro-2,7-dihydroxy-4,6-dimethoxyphenanthrene, 4, and the previously unreported 1,2,6,7-tetrahydroxy-4-methoxyphenanthrene, 5, were isolated from the South American orchid, Brasiliorchis porphyrostele. An in depth analysis of their vascular effects was performed on in vitro rat aorta rings and tail main artery myocytes. Compounds 1-4 were shown to possess vasorelaxant activity on rings pre-contracted by the ±1 receptor agonist phenylephrine, the CaV1.2 stimulator (S)-(-)-Bay K 8644, or depolarised with high K+ concentrations. However, compound 5 was active solely on rings stimulated by 25 mM but not 60 mM K+. The spasmolytic activity of compounds 1 and 4 was significantly affected by the presence of an intact endothelium. The KATP channel blocker glibenclamide and the KV channel blocker 4-aminopyridine significantly antagonised the vasorelaxant activity of compounds 4 and 1, respectively. In patch-clamp experiments, compounds 1-4 inhibited Ba2+ current through CaV1.2 channels in a concentration-dependent manner, whereas neither compound 4 nor compound 1 affected K+ currents through KATP and KV channels, respectively. The present in vitro, comprehensive study demonstrates that Brasiliorchis porphyrostele may represent a source of vasoactive agents potentially useful for the development of novel antihypertensive agents that has now to be validated in vivo in animal models of hypertension.
Langat Moses K., Ndunda Beth M., Salter Caitlin, Odusina Babatope O., Isyaka Sani M., Mas-Claret Eduard, Onocha Patricia A., Midiwo Jacob O., Nuzillard Jean-Marc, Mulholland Dulcie A. (2020) Diterpenoids from the stem bark of Croton megalocarpoides Friis & M. G. Gilbert,Phytochemistry Letters 39 pp. 1-7 Elsevier
Five previously undescribed compounds, megalocarpoidolide I (1), megalocarpoidolide J (3), 12-epi-crotonzambefuran A (4), megalocarpoidolide K (5), 1-trans-p-hydroxycoumaroyl?geranylgerani-1-ol (6) were isolated from the stem bark of Croton megalocarpoides Friis & M. G. Gilbert. The known ent-trachyloban-18-ol, megalocarpoidolide B, megalocarpoidolide C (2), megalocarpoidolide H, crotocorylifuran, 7,8-dehydrocrotocorylifuran, 1,2-dehydrocrotocorylifuran-2-one, acetyl aleuritolic acid, lupeol, N-trans-p-coumaroyl-32,42-dihydroxyphenylethylamine, dodecyl trans-ferulate and lignoceryl trans-ferulate were also isolated. The structures of the compounds were determined using NMR, IR spectroscopy and HRMS. The structure of compound 1 was determined using Logic for Structural Determination (LSD). Compounds 1, 2 and 3 that were selected for screening based on their ability to add diversity to the NCI small molecule compound collection, were evaluated against the NCI60 panel of human tumour cell lines at 10¼M level but found to be inactive.
Moodley N., Crouch N.R. Crouch, Bastida J., Mas-Claret E., Houghton P., Mulholland D.A. (2020) Organ-specific production of alkaloids from bulbs and seeds of Crinum stuhlmannii subsp. delagoense (Amaryllidaceae),South African Journal of Botany Elsevier
The phytochemistry of both bulbs and seeds of the ethnomedicinal Crinum stuhlmannii subsp. delagoense (Amaryllidaceae) was studied, sourced from a single location simultaneously. Eight alkaloids, including two previously unreported ones, N-methyl-haemanthamide (nivanine), and N-methyl-ent-delagoenine, were isolated along with lycorine, 8,9-methylenedioxophenanthridine, 6-hydroxycrinamine, 6-ethoxycrinamine, haemanthamine and hamayne. Compounds isolated were tested for acetylcholinesterase inhibition.
Moodley N., Crouch N.R., Bastida J., Mulholland D.A. (2020) Novel alkaloids and a ceramide from Brunsvigia natalensis (Amaryllidaceae) and their anti-neoplastic activity,South African Journal of Botany Elsevier
Two new 5,10b-ethanophenanthridine bridge alkaloids, 3-O-methyl?epi?vittatine, 1, and crouchinine, 2, (2R)-7-hydroxyflavan, 3, and a novel ceramide, brunsceramide, were isolated from the bulbs of Brunsvigia natalensis. (2R)-7-Hydroxyflavan showed total growth inhibition in the 55?60 ppm range against the melanoma (UACC62), breast (MCF7) and renal (TK10) cancer cell lines. 3-O-Methyl?epi?vittatine, 1, and bruns-ceramide, 4, were inactive against the cell lines tested.