Henry Lowe

Cycloartanes with Anticancer Activity Demonstrate Promising inhibition of the Mrckα and Mrckβ Kinases

Aim:The role of Kinases in cancer onset and progression has made kinases a target for the control of some cancers.Recent discoveries that kinases are most effectively inhibited bysmall molecules havealso resulted in an increased search for small molecule kinase inhibitors. Cycloartanes are small molecules found in many medicinal plants including the Jamaican Ball Moss ( Tillandsia recurvata ). Recent studies onT. recurvatahave demonstrated that it possesses anticancer activity. Cycloartane -3,24,25- triol, an analog of a cycloartane identified in Ball moss was also shown to have inhibitory activity against MRCK�± kinase. This study was as such set up to determine the MRCK�±/�≤ kinase i nhibition activity of other cycloartanes in Ball Moss and their analogs. MRCK�±/�≤ kinases has been identified as an important kinase implicated in cancer onset and progression and as such a potential drug target. Methodology :Kinase inhibition activity of 6 cycloartanes was investigated using the

Petiveria alliacea L (Guinea Hen Weed) and Its Major Metabolite Dibenzyl Trisulfide Demonstrate HIV-1 Reverse Transcriptase Inhibitory Activity

Aim: The human immunodeficiency virus (HIV) remains a major public health concern despite the discovery and development of Highly Active Antiretroviral Therapies (HAART). There is as such a need to continue to search for new and effective therapies for this global pandemic. In an effort to discover new anti HIV agents, the aim of this study was to determine the anti HIV-1 activity of Petiveria alliacea and its metabolites . Methodology: The extracts of P. alliacea and dibenzyl trisulfide were screened for anti HIV-1 properties in primary peripheral blood mononuclear cells (PBMCs) infected with the HIV-1JR-CSF strain.

Plasma Cocaine Metabolite and Liver CYP450 3A4 Isoenzyme Levels as Indicators of Cocaine Dependence in Rats Treated with Nutritional Supplements

The effects that chronic cocaine administration CCA have on craving, cocaine metabolite concentrations and cytochrome P450 3A4 isoenzyme CYP450 3A4 activities in Sprague-Dawley rats following the administration of Salako Nutritional Supplements SNS were examined. Five groups of fifty rats were used to assess the effect of the SNS following CCA. Craving was analyzed for each rat using a Conditioned Place Preference protocol. Blood samples were obtained at regular intervals and used to measure cocaine plasma metabolite levels. CYP450 3A4 activity was determined in the liver. Administration of the SNS reduced craving of cocaine significantly, upon discontinuing cocaine in the rats. Blood plasma analysis showing higher benzoylecgonine equilibrium and the CYP450 3A4 levels demonstrated that the SNS possibly aided in the removal of the stored metabolites indicative of increased metabolism of cocaine, enhanced by the Supplements. Results indicate that the SNS formulation reduces craving caused by CCA by increasing the liver CYP450 3A4 activity, resulting in better plasma clearance.

Anti HIV-1 Activity of the Crude Extracts of Guaiacum officinale L. (Zygophyllaceae).

Aim: Jamaica is rich in medicinal plants. Guaiacum officinale is the “National Flower”, with reported uses in folk medicine for the treatment of various conditions including inflammation. In our search for plants with anticancer and anti-infective properties, we evaluated Guaiacum officinale for activity against HIV-1. Methodology: The leaf, seed and twig extracts of G. officinale were screened for anti HIV-1 properties in primary peripheral blood mononuclear cells (PBMCs) infected with the reference HIV-1 BaL strain. Results: All the tested extracts inhibited HIV-1 p24 production by infected cells, with EC50 concentrations of 22.35μg/ml, 23.42μg/ml and 25.04μg/ml, respectively for the leaf, seed and twig extracts. As comparison, Betulinic acid had an EC50 value of 27.50μg/ml. The tested extracts had IC50/EC50 selectivity index (SI) values of ≥ 3, which compared favorably to Betulinic acid SI value of 1.09. Conclusion: The results of this study suggest that extracts of G. officinale may provide leads for the discovery of new drug agents against HIV-1. Short Research Article European Journal of Medicinal Plants, 4(4): 483-489, 2014 484

Unearthing the medicinal properties of Tillandsia recurvata (Ball Moss): a mini review.

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Abstract 1754: Cycloartane anticancer activity

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA New cancer diagnoses remain on the rise despite significant improvements in early detection and treatment. The role of Kinases in cancer onset and progression has made kinases a target for the control of some cancers. The MRCKα/β kinases have recently been implicated in cancer onset and progression and as such could serve as a potential drug target. The discovery that kinases are most effectively inhibited by small molecules has also resulted in an increased search for small molecule kinase inhibitors. Cycloartanes are small molecules found in many medicinal plants including the Jamaican Ball Moss (Tillandsia recurvata). Recent studies on T. recurvata have demonstrated that it possesses significant anticancer activity. Cycloartane-3,24,25-triol, an analog of a cycloartane identified in Ball moss was also shown to have inhibitory activity against MRCKα kinase. This study was as such set up to determine the MRCKα/β kinase inhibition activity of other cycloartanes in Ball Moss and their analogs. The kinase inhibitory activity of 6 cycloartanes was investigated using the ligand-kinase binding assay while the WST-1 reagent assay was used to determine the antiproliferative activity of the cycloartanes against some prostate and breast cancer cell lines. Cycloart-23-ene-3,25-diol (1), Cycloartane-3,24,25-triol (2), Cycloart-25-ene-3,24-diol (3), 3,23-Dioxo-9,19-cyclolanost-24-en-26-oic acid (4), 24,25-Dihydroxycycloartan-3-one (5) inhibited the MRCKα kinase with Kd of 0.21 μM, 0.25µM, 0.36 µM, 3.0 µM, and 2.1 µM respectively. Hydroxycycloart-23-en-3-one,25, (6) showed no inhibition against the MRCKα kinase. Compounds 1, 3, 4, 5 inhibited the MRCKβ kinase with Kd of 4.7 μM, 1.10 µM, 3.2 µM, and 9.8 µM, respectively. Three of the six cycloartanes exhibited antiproliferation activity against two prostate and breast cancer cell lines each. In conclusion, cycloart-23-ene-3,25-diol (1) showed the most promising activity against the MRCKα/β kinase out of the 6 cycloartanes screened demonstrating an interesting structure activity relationship profile when compared with the other molecules. Cycloart-23-ene-3,25-diol (1) deserves further studies to determine its in vivo efficacy and safety. Citation Format: Henry Isaac Cloore Lowe, Ngeh J. Toyang, Charah Watson, Joseph Bryant. Cycloartane anticancer activity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1754. doi:10.1158/1538-7445.AM2014-1754

Abstract 4237: Inhibition of prostate cancer associated kinases by Tillandsia recurvata: A Jamaican medicinal plant

Prostate cancer is the second most common cause of death from cancer in men of all ages. The three conventional treatment options include surgery, radiation and chemotherapy. In many cases, these treatments are used in combination. Chemotherapy is however the most widely used tool in prostate cancer treatment especially where the cancer has metastasized or spread to other organs of the body. Increased resistance to current chemotherapies by prostate cancer calls for an urgent need to discover and develop new therapeutics that can slow the growth of cancer cells while having lesser side effects on the patients. The development of target therapies for the treatment of human cancers is revolutionizing the concept of cancer treatment today. This type of therapeutic approach is directed to the inhibition of molecular targets that play a pivotal role in tumor progression such as the kinases. It is only in recent years that kinases have emerged as major targets for therapeutics, particularly for cancer related therapies. Relatively few kinase inhibitors have been identified to date and efforts are being made to discover new small molecules kinase inhibitors. The Island of Jamaica is known for its rich biodiversity and abundance of medicinal plants used in folk medicines. In an effort to discover new anticancer drugs from natural products several Jamaican medicinal plants were screened for anticancer activity. The Jamaican Ball Moss (Tillandsia recurvata L.) was one such plant that exhibited potent activity against the PC-3 prostate cancer cell line in vitro (IC 50 = 2.4µg/ml) and was selected for further studies. The crude extract of Ball Moss was screened for interaction with 451 kinases. The crude Ball Moss extract showed potent kinase inhibitory activity against 5 kinases (kd = 8-14 µg/ml), of which 4 are implicated in prostate cancer growth and proliferation. The four kinases are; CSNK2A2, DRAK1, GAK and MEK5. The above results validate the folk use of this medicinal plant as well as make it a good candidate for development into a prostate cancer treatment or for the discovery of the bioactive molecules in the plant. Studies are currently underway to confirm the ability of Ball Moss to halt the growth of prostate tumors in mice as well as measure the inhibitory effects of the crude extract on the 4 related kinases in vivo. The isolation and characterization of the kinase inhibitors in ball moss is also in progress. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4237. doi:1538-7445.AM2012-4237

Abstract 5266: Anti-cancer drug development using bioactive compounds from Tillandsia recurvata in mouse models of human cancer

Background: The Jamaican Tillandsia recurvata species is being fully investigated scientifically for medicinal properties and has demonstrated powerful anti-cancer properties in vitro and in vivo. Aim The scope of this study was to investigate the anti-tumor and anti-HIV effects of Ball Moss extracts, isolates and synthesized molecules both in vivo and in vitro in five histogenic cell lines: Melanoma, Prostate, Breast, Kaposi sacrcoma and B16-Lymphoma cancer cell lines. Methodology/Experimentation: The bioactive compounds from Ball Moss were first extracted, isolated and identified. The fresh plant material was collected from power lines in Jamaica after which it was thorough washed, dried in a steam oven at 70°C, then milled and extracted with methanol (MeOH). Isolation of the bioactive compounds was done in two phases. Firstly, isolates from the crude extract was isolated by column chromatography and sub-fractions processed by bio-guided fractionation. During phase two, advanced separation technologies, including HPLC, supercritical fluid chromatography and capillary electrophoresis, were used followed by LC/MS and finally NMR. The bioassays of the isolated fractions were tested against the five different histogenic cancer cell lines in vitro using trypan blue exclusion and 3H Thymidine incorporation assays. The in vivo studies included 10 nude mice per group and using the crude form of the extract at 10mg per mice per day for 7 days. The extracts were tested in vivo against the same tumor cell lines and compared to controls administered normal saline treatment. Results: Utilizing the bioassay-guided fractionation process, the bioactive moiety was isolated at 98% purity. This purified compound tested in vitro demonstrated to be highly effective at a rate of in excess of 95% cell kill in the 5 different histogenic cell lines. The in vivo studies were equally impressive utilizing the crude extract. All 5 different tumors responded to the treatment by reducing the tumor size from 0.4mm X 0.4mm to almost non-existent on gross examinations of tumor cell lines (see images below). While the histological assessment using immuno-histochemical staining revealed that 90 to 95% of the tumors had undergone cell death cell which was due to apoptosis. There were no observations in the animals of any indications of any toxicity in the in vivo studies. The representative sample of gross tumors from a xenograft of the growth of Kaposi Sarcoma in nude mice and the histological results showing extensive necrosis following the use of Ball Moss extract and the immune-histochemistry, demonstrated cell death by apoptosis. It is to be noted that the treatments of all 5 cancer cell demonstrated the same level of bioactivity. Discussion/Conclusion: The findings from this study strongly indicate that these newly extracted compounds have significant anti-cancer / anti-HIV properties. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5266. doi:1538-7445.AM2012-5266