Ehrlich II –2nd World Conference on Magic Bullets

Genetic Variant of KIF6 Predicts both Increased Risk for Coronary Events and Greater Benefit from Statin Therapy: an Overview of Genetic Studies of the CARE, WOSCOPS, and PROVE IT - TIMI 22 Trials IAKOUBOVA OA^1, SABATINE MS^2,^,ROWLAND CM^1, TONG CH^1, CAMPOS H^4, PFEFFER MA^2, KIRCHGESSNER TG^5, PLOUGHMAN LM^5, SIMONSEN KL^5, RANADE K^5, WHITE TJ^1, PACKARD CJ^3, SACKS FM^2,4, SHEPHERD J^3, DEVLIN JJ^1 AND BRAUNWALD E^2 ^1Celera, Inc., Alameda, CA, USA, ^2Brigham & Women’s Hospital, Harvard Medical School, Boston, USA, ^3University of Glasgow and Royal Infirmary, Glasgow, UK., ^4Harvard School of Public Health, Boston, USA, ^5Pharmaceutical Research Institute, Bristol-Myers Squibb, Princeton, USA Background: Statins are the drugs of choice for primary and secondary prevention of coronary heart disease; however, response to statin therapy varies dramatically between individuals. Aim: To identify and validate genetic polymorphisms that are associated with risk of coronary events and differential response to statin therapy. Methods: We used Cox proportional hazards models that adjusted for traditional risk factors to investigate the effect of pravastatin therapy versus placebo by KIF6 719Arg carrier status and the association between KIF6 719Arg carrier status and coronary events in the CARE and WOSCOPS trials and to investigate the effect of high-dose atorvastatin therapy versus standard-dose pravastatin therapy in the PROVE IT–TIMI 22 trial. Results: The 719Arg variant of Trp719Arg (rs20455), a polymorphism in kinesin-like protein 6, was associated with greater risk of coronary events and greater benefit from pravastatin versus placebo. In placebo–treated patients, carriers of the KIF6 719Arg allele (59% of CARE and WOSCOPS) had a hazard ratio of 1.50 (95% CI 1.05 - 2.15) in CARE and an odds ratio of 1.55 (95% CI 1.14 - 2.09) in WOSCOPS. Among 719Arg carriers, the absolute risk reduction by pravastatin was 4.89% (95% CI 1.81 - 7.97) in CARE and 5.49% (95% CI 3.52 - 7.46) in WOSCOPS. In contrast, no significant risk reduction was observed among noncarriers. In PROVE IT–TIMI22, benefit from high-dose, compared with standard-dose, statin therapy was significantly greater in the 59% of the cohort who were carriers (hazard ratio 0.59, 95% CI 0.45 - 0.77) than in noncarriers (hazard ratio 0.94, 95% CI 0.70 - 1.27); p=0.018 for interaction between 719Arg carrier status and treatment. Absolute risk reduction was 10.0% in carriers versus 0.8% in noncarriers. Conclusions: 1) Carriers of the KIF6 719Arg allele are at increased risk of coronary events, and pravastatin therapy substantially reduces that risk. 2) Carriers of 719Arg receive significantly greater benefit from high-dose statin therapy than do noncarriers. 3) In all three trials, noncarriers of 719Arg (representing over 40% of the populations) did not benefit from statin therapy. Since statin therapy may obscure benefits of other cardiovascular drugs, demonstrating the benefit of new cardiovascular compounds over highly potent statins may be more successful in statin non-responders identified by the KIF6 polymorphism.	Oral Cancer: Molecular Pathogenesis and Novel Therapeutic Approach IAMAROON A., KRISANAPRAKORNKIT S. Chiang Mai University, Chiang Mai, Thailand Background: Aberration of signal transducers in PI3K/AKT pathway has been found in many human cancers including oral cancer and may play a critical role in carcinogenesis of those cancers. Advanced research on the treatments of oral cancer using novel agents targeting on PI3K/AKT signaling pathway are investigated in many laboratories with promising results. The objectives of the present study were (1) to investigate protein expression of pan AKT and its phosphorylated form, p-AKT, in oral squamous cell carcinoma (OSCC) tissues of 20 Thai patients, (2) to analyze mRNA expression of three isoforms of AKT; AKT-1, -2, and -3 and protein expression of pan AKT, AKT-1, and AKT-2 in OSCC cell lines and human oral keratinocytes (HOK), and (3) to analyze protein expression of vimentin and E-cadherin in OSCC and HOK cell lines. Methods: The expression of pan AKT and p-AKT in OSCC tissues was studied by immunohistochemistry. The mRNA expression of AKT-1, -2, and -3 in OSCC cell lines and HOK was analyzed by RT-PCR and the protein expression of pan AKT, AKT-1, AKT-2, vimentin, and E-cadherin was studied by Western blot assay. Results: The results showed that pan AKT and p-AKT were overexpressed in 95% and 100% of OSCC cases, respectively. We observed more intense expression of pan AKT and p-AKT at the invasive fronts of some OSCC tissues. Pan AKT protein was also overexpressed in all OSCC cell lines in comparison with HOK. Interestingly, AKT-1 and -2 mRNA of OSCC cell lines were only constitutively expressed in comparison with HOK. AKT-3 mRNA appeared to be minimally expressed in OSCC cell lines and HOK. The Western blot analysis revealed that AKT-2 but not AKT-1 was overexpressed. Additionally, vimentin was upregulated while E-cadherin was downregulated. Conclusions: These findings suggested that overexpression of pan AKT particularly AKT-2 and p-AKT may be involved with OSCC carcinogenesis and post-transcriptional modification of the expression of AKT isoforms in OSCC may occur. In addition, OSCC cells may undergo epithelial-mesenchymal transition since their epithelial maker (E-cadherin) was reduced whereas their mesenchymal marker (vimentin) was increased.
Peroral Colon-Specific Delivery of Insulin Based on Novel Acrylic-Terpolymer Microcapsular Devices ICHIKAWA H^1, ARIMOTO M^1,2, FUKURMORI Y^1 ^1Kobe Gakuin Univ., Kobe, Japan; ^2Arfresa Pharma Corporation, Osaka, Japan. Background: Recently, colon has attracted much attention as a potentially delivery site for perorally administered peptide-based drugs. In this context, we have developed novel delayed-release microcapsules (MCs) with a membrane of newly synthesized acrylic terpolymers as a prototype of colon-specific delivery device for peptide-based drugs. Aims: 1) To prepare insulin-containing MCs. 2) To evaluate stability, in vitro release and in vivo absorption behavior of the microencapsulated insulin. Methods: An aqueous colloidal dispersion of terpoly(ethyl acrylate/methyl methacrylate/2-hydroxyethyl methacrylate) with molar ratio of 95:85:40 was synthesized by emulsion polymerization. The MCs composed of a lactose core (90–105 microns), a layer of bovine pancreatic insulin with a protease inhibitor (bacitracin) and an absorption enhancer (sodium glycocholate), and a release-delaying coat of the terpolymers were prepared by the air-suspension spray coating process. The obtained MCs were heat-cured at 40˚C for 6h and then subjected to stability assay by an HPLC method, release test using a paddle method and absorption study of insulin after peroral administration to gastro-physiology-regulated beagle dogs. Results: The MCs with mass median diameters of 175–226 microns were obtained at the yield of 86–93%. Degradation of insulin during the spray-coating process and post-thermal curing was only few % when the process temperatures were set to below 40˚C. The heat-cured MCs showed delayed-release of insulin in a pH-independent manner. The lag-time of drug-release could be controlled by altering the coat thickness of terpolymers. Peroral administration of the MCs with 6-h lag-time to the beagle dogs revealed a significant reducing effect of blood glucose level (the pharmacological abailability was estimated to be 5.1%) while that of the MCs with 3-h lag-time did not. Moreover, the microencapsulated insulin was found to be stable and its release profile was not changed significantly even after the storage at 4˚C for 2 years. Conclusions: 1) Microencapsulation of insulin with the terpolymers was possible without significant degradation of insulin. 2) The microencapsulated insulin was stable for 2 years, released pH-independently in a delayed mannar, and effective to enhance in vivo insulin absorption thorugh the colon.	Antiplasmodial and Immunomodulating Activity of Some Sudanese Herbal Medicine with emphasis on Pristimerin as Antiplasmodial and Antileishmanial Agent Idris A elT M^1,2, satti G M H. ^1, Theander T^3, ChristensenS B. ^4 , Khalid S a.^5 ^1Faculty of Medicine, King Fahad Medical City, Kingdom of Saudi Arabia ^2Dept. of Biochemistry, Faculty of Medicine, University of Gezira, Sudan ^3Panum Institute, University of Copenhagen, Copenhagen, Denmark ^4Department of Medicinal Chemistry, Natural Products Research, The Danish University of Pharmaceutical Sciences, DK-2100 Copenhagen, Denmark ^5Dept. of Pharmacognosy, Faculty of Pharmacy, University of Khartoum, Sudan Background: The Sudan is being the largest country in Africa, covering an area of one million square miles with the different metrological and polyethnic, with a diverse flora. Most people in rural areas rely on traditional medicine for the treatment of many infectious diseases. Objectives: WHO has recently advocated the use of traditional medicine where appropriate health services become inaccessible, therefore, the study aims to investigate the potential antiplsmodial, antleishmanial activity of some medicinal plants and to detect their effect on human lymphocytes proliferation which may imply the ability to potentiate the human immune system. Material and methods: Forty-nine plant parts representing 26 species from 15 families were extracted and screened for their activity on chloroquine sensitive strains 3D7 and Dd2. Plants were collected according to their traditional use and / or to their taxonomical affiliation to their families that had been reported to have antimalarial activity. Results: Thirty-four methanol extracts (59%) exhibited significant activity against 3D7 with IC50 values ≤ 50 µg/ml, while twenty-one extract (57%) showed antiplasmodial activity on Dd2 with IC50 values ≤ 50 µg/ml. On the other hand, thirteen extracts (22%) and ten extracts (18%) only showed an activity with IC50 values ≤ 5 µg/ml on 3D7 and Dd2; respectively. Human lymphocytes treated with the most of extracts demonstrated a minimum level of toxic inhibitory effect at concentration ≥ 100µg, whereas Sonchous cornatus, Balanites aegyptiaca, Acacia nilotica and Tamarindus indica enhanced lymphocytes proliferation. Bioactivity directed fractionation of the chloroform extract of the root bark of Maytenus senegalensis resulted in the isolation and characterization of the quinonemethide triterpene, (20α)-3-hydroxy-2-oxo-24-nor-friedela-1(10),3,5,7-tetraen-carboxylic acid - (29)- methylester (pristimerin). The structure was elucidated by spectroscopic techniques. The in vitro antiplasmodial activity of the isolated compound against chloroquine-resistant strain (Dd2) of Plasmodium falciparum was IC50 = 0.5μg/ml and its in vitro antileishmanial activity performed on promostigotes of Leishmania major was IC50 = 6.8 ± 0.8 μg/ml while the cytotoxicity on lymphocyte proliferation model was detected at IC50= 6.8 ± 0.8 μg/ml. Conclusion: The promising response of Acacia nilotica and Mayenus senegalensis conclude that some Sudanese plants used in traditional medicine possess a potent antimalarial activity with minor effects on lymphocytes proliferation. These plants have been subjected to long-term clinical trials in folk medicine and hence we propose that these plants should be further investigated.