Volodymyr Samoylenko

Antiparasitic, nematicidal and antifouling constituents from Juniperus berries

A bioassay‐guided fractionation of Juniperus procera berries yielded antiparasitic, nematicidal and antifouling constituents, including a wide range of known abietane, pimarane and labdane diterpenes. Among these, abieta‐7,13‐diene (1) demonstrated in vitro antimalarial activity against Plasmodium falciparum D6 and W2 strains (IC50 = 1.9 and 2.0 µg/mL, respectively), while totarol (6), ferruginol (7) and 7β‐hydroxyabieta‐8,13‐diene‐11,12‐dione (8) inhibited Leishmania donovani promastigotes with IC50 values of 3.5–4.6 µg/mL. In addition, totarol demonstrated nematicidal and antifouling activities against Caenorhabditis elegans and Artemia salina at a concentration of 80 µg/mL and 1 µg/mL, respectively. The resinous exudate of J. virginiana afforded known antibacterial E‐communic acid (4) and 4‐epi‐abietic acid (5), while the volatile oil from its trunk wood revealed large quantities of cedrol (9). Using GC/MS, the two known abietanes totarol (6) and ferruginol (7) were identified from the berries of J. procera, J. excelsa and J. phoenicea. Copyright

Indolizidine, antiinfective and antiparasitic compounds from Prosopis glandulosa var. glandulosa.

A new potent antiinfective and antiparasitic 2,3-dihydro-1H-indolizinium chloride (1) was isolated from Prosopis glandulosa var. glandulosa. Three additional new (2-4) and one known (5) indolizidines were also isolated, and the dihydrochloride salts of 1-3 (compounds 6, 7, and 8) were prepared. Structures were determined by 1D and 2D NMR and mass spectra. Compound 1 showed potent in vitro antifungal activity against Cryptococcus neoformans and Aspergillus fumigatus (IC(50) values = 0.4 and 3.0 microg/mL, respectively) and antibacterial activity against methicillin-resistant Staphylococcus aureus and Mycobacterium intracellulare (IC(50) values of 0.35 and 0.9 microg/mL, respectively). The remarkable in vitro fungicidal activity of 1-4 against C. neoformans (MFCs = 0.63-1.25 microg/mL) and 2, 3, and 5 against A. fumigatus (MFCs = 0.63-2.5 microg/mL) were similar to amphotericin B, but >2-4-fold more potent than 6-8. Prosopilosidine (1) showed potent in vivo activity at 0.0625 mg/kg/day/ip for 5 days in a murine model of cryptococcosis by eliminating approximately 76% of C. neoformans infection from brain tissue compared to approximately 83% with amphotericin B at 1.5 mg/kg/day. Compounds 1 and 4 exhibited potent activity and high selectivity index (SI) values against chloroquine-sensitive (D6) and chloroquine-resistant (W2) strains of Plasmodium falciparum, with IC(50) values of 39 and 95 ng/mL and 42 and 120 ng/mL, respectively (chloroquine, IC(50) = 17 and 140 ng/mL). Prosopilosine (1) also showed in vivo antimalarial activity, with an ED(50) value of approximately 2 mg/kg/day/ip against Plasmodium berghei-infected mice after 3 days of treatment.

Antiparasitic and antimicrobial indolizidines from the leaves of Prosopis glandulosa var. glandulosa.

A new indolizidine alkaloid, named Δ¹,⁶-juliprosopine (1), together with previously known indolizidine analogs (2- 6), was isolated from the leaves of Prosopis glandulosa var. glandulosa, collected from Nevada, USA; while two other known indolizidines, juliprosopine (6) and juliprosine (7), were isolated from P. glandulosa leaves collected in Texas, USA. The structures of compound 1 and 7 were determined using a combination of NMR and MS techniques. Compound 7 exhibited potent antiplasmodial activity against Plasmodium falciparum D6 and W2 strains with IC (50) values of 170 and 150 ng/mL, respectively, while 1 was found to be less active (IC₅₀ values 560 and 600 ng/mL, respectively). Both compounds were devoid of VERO cells toxicity up to a concentration of 23 800 ng/mL. The antileishmanial activity of indolizidines was evaluated against Leishmania donovani promastigotes, axenic amastigotes, and amastigotes in THP1 macrophage cultures. When tested against macrophage cultures, the tertiary bases (1, 3, 6) were found to be more potent than quaternary salts (2, 5, 7), displaying IC₅₀ values between 0.8-1.7 µg/mL and 3.1-6.0 µg/mL, respectively. In addition, compound 7 showed potent antifungal activity against Cryptococcus neoformans and antibacterial activity against Mycobacterium intracellulare, while 1 was potent only against C. neoformans and weakly active against other organisms.

Composition, Standardization and Chemical Profiling of Banisteriopsis caapi, a Plant for the Treatment of Neurodegenerative Disorders Relevant to Parkinson’s Disease

ETHNOPHARMACOLOGICAL RELEVANCE Banisteriopsis caapi, a woody vine from the Amazonian basin, is popularly known as an ingredient of a sacred drink ayahuasca, widely used throughout the Amazon as a medicinal tea for healing and spiritual exploration. The usefulness of Banisteriopsis caapi has been established for alleviating symptoms of neurological disorders including Parkinsons disease. AIM OF THE STUDY Primary objective of this study was to develop the process for preparing standardized extracts of Banisteriopsis caapi to achieve high potency for inhibition of human monoamine oxidases (MAO) and antioxidant properties. The aqueous extracts prepared from different parts of the plant collected from different geographical locations and seasons were analyzed by HPLC for principal bioactive markers. The extracts were simultaneously tested in vitro for inhibition of human MAOs and antioxidant activity for analysis of correlation between phytochemical composition of the extracts and bioactivities. MATERIALS AND METHODS Reversed-phase HPLC with photodiode array detection was employed to profile the alkaloidal and non-alkaloidal components of the aqueous extract of Banisteriopsis caapi. The Banisteriopsis caapi extracts and standardized compositions were tested in vitro for inhibition of recombinant preparations of human MAO-A and MAO-B. In vitro cell-based assays were employed for evaluation of antioxidant property and mammalian cell cytotoxicity of these preparations. RESULTS Among the different aerial parts, leaves, stems/large branches and stem bark of Banisteriopsis caapi, HPLC analysis revealed that most of the dominant chemical and bioactive markers (1, 2, 5, 7-9) were present in high concentrations in dried bark of large branch. A library of HPLC chromatograms has also been generated as a tool for fingerprinting and authentication of the studied Banisteriopsis caapi species. The correlation between potency of MAO inhibition and antioxidant activity with the content of the main active constituents of the aqueous Banisteriopsis caapi extracts and standardized compositions was established. Phytochemical analysis of regular/commercial Banisteriopsis caapi dried stems, obtained from different sources, showed a similar qualitative HPLC profile, but relatively low content of dominant markers 1, 2, 7, and 9, which led to decreased MAO inhibitory and antioxidant potency compared to Banisteriopsis caapi Da Vine. CONCLUSION The ethnopharmacological use of bark of matured stem/large branch of Banisteriopsis caapi as well as whole matured stem is supported by the results obtained in this investigation. Among various constituents of Banisteriopsis caapi, harmine (7), harmaline (6) and tetrahydroharmine (5) are responsible for MAO-A inhibition, while two major proanthocyanidines, epicatechin (8) and procyanidine B2 (9) produce antioxidant effects. The compounds 1-9 can serve as reliable markers for identification and standardization of Banisteriopsis caapi aerial parts, collected in different seasons and/or from different geographical regions.

Antimalarial quassinoids: past, present and future

This review is a compilation of the investigations reported to date on the sources, isolation, chemistry and antimalarial activities of natural quassinoids and their synthetic and semisynthetic analogs. It also provides an analysis of the in vitro structure–activity relationship of quassinoids for further evaluation in animal models. The introduction of non-nitrogenous antimalarial drugs has created a new era of malaria chemotherapy to treat Plasmodium falciparum strains that are resistant to existing nitrogenous drugs and the rising incidence of the deadly cerebral malaria. Many antimalarial quassinoids are discovered from simaroubaceous plants that are used traditionally to treat fever and malaria, thereby reiterating the critical role of ethnopharmacology as a rich source of novel drug discovery.

Chemical and Biological Study of Flueggea virosa Native to Saudi Arabia

Flueggea virosa (Roxb. ex Willd.) Royle (Phyllanthaceae), also known as Chinese waterberry, is a shrub up to 4 m high growing wild in tropical Africa, Middle East, tropical Asia, Japan, Australia, and Polynesia and can also be grown domestically. It has many recorded uses in Chinese herbal medicine. The plant has been used for the treatment of fever, malaria, sexual dysfunction, pain, diabetes, epilepsy, snakebite, venereal disease, rheumatism, arrhythmia, rash, diarrhea, pneumonia, cough, hepatitis, and HIV-related illness, and as a contraceptive [1, 2]. The methanol and water extracts of the plant have been reported to possess a number of biological activities such as antiplasmodial, trypanocidal, and antiarrhythmic [3, 4]. Previous phytochemical studies of F. virosa showed the presence of triterpenoids [2], tannins, flavonoids, saponins [5, 7], resins, glycosides, glycerin carbohydrates, anthraquines, steroids [5], alkaloids [6, 8], cardiac glycosides [5, 6], and anthraquinones [6]. In our phytochemical investigation of this plant [9], a total of 14 compounds were isolated from the ethanolic extract of the aerial parts of F. virosa, collected from the Southern part of Saudi Arabia, in June 2000. The plant (air-dried crushed aerial parts, 1 kg) was macerated with 95% EtOH at room temperature overnight (1 L each 3). The dried crude EtOH extract (40 g) was dissolved in 0.1 N HCl (1 L; pH 1) and partitioned with n-hexane and n-BuOH (500 mL 3) successively. The n-BuOH fraction was subjected to flash column chromatography (CC) over silica gel to afford bergenin [white crystals, mp 236–238 C (1, 10 g)], menisdaurin [brown solid, mp 172–173 C (2, 1 g)] and amiroside [brown solid, mp 176–178 C (3, 25 mg)] [9, 10]. The defatted aqueous acidic fraction was basified with NH4OH (pH 11) and partitioned with EtOAc. The EtOAc fraction (3 g) was subjected to flash chromatography on silica gel with further purification using reverse phase CC over C-18 silica gel (MeCN–H2O, 9:1) to afford ent-phyllanthidine [yellow white crystals, mp 158–160 C (4, 30 mg)] [11], securinine [white powder, mp 140–142 C (5, 8 mg)], securinol [white powder, mp 136–137 C (6, 6 mg)], viroallosecurinine [dark yellow solid, mp 130–132 C (7, 40 mg)] [12], flavonoid rutin [yellow solid, mp 194–195 C (8, 40 mg)] [13], gallocatechin (9) and epi-gallocatechin [white solid, mp 188–190 C] (10, mixture, 12 mg) [14], 2,2 ,5,5 -tetrahydroxybiphenyl [white solid, mp 144–145 C (11, 12 mg)], ethylmanopyranoside [white solid, mp 154–156 C (12, 132 mg)], ethyl glucopyranoside [white powder, mp 163–164 C (13, 9 mg)] [9], and the triterpene precursor squalene [oil (14, 20 mg)] [15]. The identity of all the isolated compounds was confirmed by direct comparison with authentic samples, as well as their physical and spectral data (UV, IR, NMR and MS) with those published in the literature. This is the first report of the cyanogenetic glucosides menisdaurin (2) and amiroside (3) isolated from the genus Flueggea, as well as from the plant family Phyllanthaceae. They were previously reported from Ilex aquifolium and Guazuma ulmifolia [16].

Efficacy of Prosopilosidine from Prosopis glandulosa var. glandulosa against Cryptococcus neoformans Infection in a Murine Model

In this study, 2,3-dihydro-1H-indolizinium alkaloid-prosopilosidine (PPD), that was isolated from Prosopis glandulosa, was evaluated against C. neoformans in a murine model of cryptococcosis. In vitro and in vivo toxicity of indolizidines were also evaluated. Mice were infected via the tail vein with live C. neoformans. Twenty-four hours post-infection, the mice were treated with PPD once a day (i.p.) or twice a day (bid) orally, or with amphotericin B (Amp B) intraperitoneally (IP), or with fluconazole (Flu) orally for 5 days. The brains of all of the animals were aseptically removed and the numbers of live C. neoformans were recovered. In vitro toxicity of indolizidine alkaloids was determined in HepG2 cells. PPD showed to be potent in vivo activity against C. neoformans at a dose of 0.0625 mg/kg by eliminating ~76% of the organisms compared to ~83% with Amp B (1.5 mg/kg). In addition, PPD was found to be equally efficacious, but less toxic, at either 0.125 or 0.0625 mg/kg compared to Amp B (1.5 mg/kg) when it was administered bid (twice a day) by an i.p. route. When tested by an oral route, PPD (10 mg/kg) showed potent activity in this murine model of cryptococcosis with ~82% of organisms eliminated from the brain tissue, whereas Flu (15 mg/kg) reduced ~90% of the infection. In vitro results suggest that quaternary indolizidines were less toxic as compared to those of tertiary bases. PPD (20 mg/kg) did not cause any alteration in the plasma chemistry profiles. These results indicated that PPD was active in eliminating cryptococcal infection by oral and i.p. routes at lower doses compared to Amp B. or Flu.