Alene McCoy is presenting her thesis proposal on characterizing angiotensin IV analogs with potential therapeutic applications for cancer and dementia. She discusses how angiotensin IV binds to the AT4 receptor, which is involved in cognition and memory. Her research has found that analogs like PNB-0718 and PNB-0405 inhibit cancer cell growth and c-Met signaling. PNB-0408 enhances cognitive abilities by inducing long-term potentiation in hippocampal neurons. Her future aims are to further characterize the pharmacokinetic profiles and blood-brain barrier penetration of these analogs.
The Expanding Reach of the Designer Drug Movement in 2011: Challenges for For...NMS Labs
This presentation considers the latest intelligence on what drugs are out in the U.S. grey market of products being sold as novelties, legal highs, “Bath Salts” and research chemicals, including an update on the latest trends in synthetic cannabinoid use and detection.
The proliferation of designer drugs in the last two years has made a remarkable change to the landscape of forensic toxicology and drug identification. The scope of compounds that require detection and measurement has grown from a few drugs that needed to be targeted in specific cases, to a wide range of esoteric compounds that arguably need to be included in general drug screens for forensic purposes. The growth continues as the industry that has built up around recreational drug manufacture adjusts in an attempt to stay one step ahead of the law.
The presentation reviews the general chemical drug classes encountered in forensic toxicology and chemistry casework, including mephedrone, methylone and MDPV, recently scheduled by the US DEA, and related the cathinones, 2C compounds, tryptamines, and pyrovalerones. This includes a survey of the latest published research, and a review of resources for analytical testing and standards.
Designer Drugs Testing Solutions for EmployersNMS Labs
Designer Drugs Testing Solutions for Employers
Presented February 1, 2012
by Dr. Barry K. Logan, PhD, DABFT
NMS Labs National Director of Forensic Services
The Expanding Reach of the Designer Drug Movement in 2011: Challenges for For...NMS Labs
This presentation considers the latest intelligence on what drugs are out in the U.S. grey market of products being sold as novelties, legal highs, “Bath Salts” and research chemicals, including an update on the latest trends in synthetic cannabinoid use and detection.
The proliferation of designer drugs in the last two years has made a remarkable change to the landscape of forensic toxicology and drug identification. The scope of compounds that require detection and measurement has grown from a few drugs that needed to be targeted in specific cases, to a wide range of esoteric compounds that arguably need to be included in general drug screens for forensic purposes. The growth continues as the industry that has built up around recreational drug manufacture adjusts in an attempt to stay one step ahead of the law.
The presentation reviews the general chemical drug classes encountered in forensic toxicology and chemistry casework, including mephedrone, methylone and MDPV, recently scheduled by the US DEA, and related the cathinones, 2C compounds, tryptamines, and pyrovalerones. This includes a survey of the latest published research, and a review of resources for analytical testing and standards.
Designer Drugs Testing Solutions for EmployersNMS Labs
Designer Drugs Testing Solutions for Employers
Presented February 1, 2012
by Dr. Barry K. Logan, PhD, DABFT
NMS Labs National Director of Forensic Services
2014 lecture next generation metabolic screening - Marrakech Ron Wevers
Next Generation Metabolic Screening is a novel technique that can be applied to body fluids as urine, plasma or cerebrospinal fluid for the diagnosis of inborn errors of metabolism. The technique gives a holistic view on metabolism (metabolomics) and uses LC_Qtof mass spectrometry. The technique was developed in Nijmegen, The Netherlands in the group of Prof Ron Wevers (ron.wevers@radboudumc.nl)
Dr. Robert Langer - Simposio Internacional 'Terapias oncológicas avanzadas'Fundación Ramón Areces
Los días 15 y 16 de octubre de 2014, la Fundación Ramón Areces y la Real Academia Nacional de Farmacia, en colaboración con la Fundación de la Innovación Bankinter, reunieron en Madrid a algunos de los mayores expertos mundiales en nuevas terapias contra el cáncer. El Simposio Internacional, coordinado por la profesora y académica María José Alonso, analizó el momento actual de la lucha contra esta enfermedad. También fue un punto de encuentro para científicos de los más innovadores institutos de investigación en oncología, quienes debatieron sobre tres grandes temas: la Medicina Personalizada contra el cáncer, los nanomedicamentos en la terapia del cáncer y las terapias basadas en la inmunomodulación.
Gold Standard Physiological Measurements and Novel Drug Delivery Methods - Se...InsideScientific
A 2-part webinar for scientists interested in novel drug delivery methods for basic research, drug discovery and development. Learn about novel infusion technologies and how challenges in physiological monitoring and drug delivery are being overcome by implantable and programmable devices.
Session 1: The harmony of gold standard physiological monitoring with novel infusion technology: Better quality of science in preclinical models ranging from mice to marmosets.
Presenter: Christian Schnell, Novartis AG, Basel, Switzerland
Mr. Schnell presents data from multiple species that show how differences in blood pressure, heart rate and acute stress impact drug activity and how dosing a pre-trained animal can influence hemodynamic parameters.
Survival of Esophageal Cancer Patients was Significantly Superior in Comparison with Cardioesophageal Cancer Patients after Surgery
Kshivets Oleg Surgery Department, Roshal Hospital, Moscow, Russia
OBJECTIVE: This study aimed to determine localization influence of tumor for 5-year survival (5YS) of esophageal (EC) or cardioesophageal (CC) cancer patients (ECP, CEP) after complete en block (R0) esophagogastrectomies (EG) through left/right thoracoabdominal incision.
METHODS: We analyzed data of 543 consecutive patients (age=56.4±8.8 years; tumor size=6±3.5 cm) radically operated (R0) and monitored in 1975-2019 (m=405, f=138; ECP=259, CEP=284; esophagogastrectomies (EG) Garlock=280, EG Lewis=263, combined EG with resection of pancreas, liver, diaphragm, aorta, VCS, colon transversum, lung, trachea, pericardium, splenectomy=151; adenocarcinoma=308, squamous=225, mix=10; T1=126, T2=114, T3=178, T4=125; N0=275, N1=69, N2=199; G1=157, G2=139, G3=247; early EC=107, invasive=436; only surgery=420, adjuvant chemoimmunoradiotherapy-AT=123: 5-FU+thymalin/taktivin+radiotherapy 45-50Gy). Multivariate Cox modeling, clustering, SEPATH, Monte Carlo, bootstrap and neural networks computing were used to determine any significant dependence.
RESULTS: Overall life span (LS) was 1892.4±2241 days and cumulative 5-year survival (5YS) reached 51.9%, 10 years – 45.7%, 20 years – 33.5%. 183 ECP lived more than 5 years (LS=4311±2419.7 days), 98 ECP – more than 10 years (LS=5903.4±2299.4 days). 224 died because of EC/CC (LS=629.2±320.1 days). 5YS of ECP (67.3%, LS=2605±2628.9 days) was significantly superior in comparison with CEP (36.4%, LS=1242.6±1558.5 days) (P=0.00000 by log-rank test). AT significantly improved 5YS (68.2% vs. 48.5%) (P=0.00033 by log-rank test). Cox modeling displayed that 5YS of ECP/CEP significantly depended on: phase transition (PT) N0—N12 in terms of synergetics, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), T, G, histology, age, AT, localization, blood cells, prothrombin index, coagulation time, residual nitrogen, blood group, Rh, glucose, protein (P=0.000-0.008). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and healthy cells/CC (rank=1), PT early-invasive EC (rank=2), PT N0—N12 (rank=3), erythrocytes/CC (4), thrombocytes/CC (5), stick neutrophils/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), eosinophils/CC (9), leucocytes/CC (10), monocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
2014 lecture next generation metabolic screening - Marrakech Ron Wevers
Next Generation Metabolic Screening is a novel technique that can be applied to body fluids as urine, plasma or cerebrospinal fluid for the diagnosis of inborn errors of metabolism. The technique gives a holistic view on metabolism (metabolomics) and uses LC_Qtof mass spectrometry. The technique was developed in Nijmegen, The Netherlands in the group of Prof Ron Wevers (ron.wevers@radboudumc.nl)
Dr. Robert Langer - Simposio Internacional 'Terapias oncológicas avanzadas'Fundación Ramón Areces
Los días 15 y 16 de octubre de 2014, la Fundación Ramón Areces y la Real Academia Nacional de Farmacia, en colaboración con la Fundación de la Innovación Bankinter, reunieron en Madrid a algunos de los mayores expertos mundiales en nuevas terapias contra el cáncer. El Simposio Internacional, coordinado por la profesora y académica María José Alonso, analizó el momento actual de la lucha contra esta enfermedad. También fue un punto de encuentro para científicos de los más innovadores institutos de investigación en oncología, quienes debatieron sobre tres grandes temas: la Medicina Personalizada contra el cáncer, los nanomedicamentos en la terapia del cáncer y las terapias basadas en la inmunomodulación.
Gold Standard Physiological Measurements and Novel Drug Delivery Methods - Se...InsideScientific
A 2-part webinar for scientists interested in novel drug delivery methods for basic research, drug discovery and development. Learn about novel infusion technologies and how challenges in physiological monitoring and drug delivery are being overcome by implantable and programmable devices.
Session 1: The harmony of gold standard physiological monitoring with novel infusion technology: Better quality of science in preclinical models ranging from mice to marmosets.
Presenter: Christian Schnell, Novartis AG, Basel, Switzerland
Mr. Schnell presents data from multiple species that show how differences in blood pressure, heart rate and acute stress impact drug activity and how dosing a pre-trained animal can influence hemodynamic parameters.
Survival of Esophageal Cancer Patients was Significantly Superior in Comparison with Cardioesophageal Cancer Patients after Surgery
Kshivets Oleg Surgery Department, Roshal Hospital, Moscow, Russia
OBJECTIVE: This study aimed to determine localization influence of tumor for 5-year survival (5YS) of esophageal (EC) or cardioesophageal (CC) cancer patients (ECP, CEP) after complete en block (R0) esophagogastrectomies (EG) through left/right thoracoabdominal incision.
METHODS: We analyzed data of 543 consecutive patients (age=56.4±8.8 years; tumor size=6±3.5 cm) radically operated (R0) and monitored in 1975-2019 (m=405, f=138; ECP=259, CEP=284; esophagogastrectomies (EG) Garlock=280, EG Lewis=263, combined EG with resection of pancreas, liver, diaphragm, aorta, VCS, colon transversum, lung, trachea, pericardium, splenectomy=151; adenocarcinoma=308, squamous=225, mix=10; T1=126, T2=114, T3=178, T4=125; N0=275, N1=69, N2=199; G1=157, G2=139, G3=247; early EC=107, invasive=436; only surgery=420, adjuvant chemoimmunoradiotherapy-AT=123: 5-FU+thymalin/taktivin+radiotherapy 45-50Gy). Multivariate Cox modeling, clustering, SEPATH, Monte Carlo, bootstrap and neural networks computing were used to determine any significant dependence.
RESULTS: Overall life span (LS) was 1892.4±2241 days and cumulative 5-year survival (5YS) reached 51.9%, 10 years – 45.7%, 20 years – 33.5%. 183 ECP lived more than 5 years (LS=4311±2419.7 days), 98 ECP – more than 10 years (LS=5903.4±2299.4 days). 224 died because of EC/CC (LS=629.2±320.1 days). 5YS of ECP (67.3%, LS=2605±2628.9 days) was significantly superior in comparison with CEP (36.4%, LS=1242.6±1558.5 days) (P=0.00000 by log-rank test). AT significantly improved 5YS (68.2% vs. 48.5%) (P=0.00033 by log-rank test). Cox modeling displayed that 5YS of ECP/CEP significantly depended on: phase transition (PT) N0—N12 in terms of synergetics, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), T, G, histology, age, AT, localization, blood cells, prothrombin index, coagulation time, residual nitrogen, blood group, Rh, glucose, protein (P=0.000-0.008). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and healthy cells/CC (rank=1), PT early-invasive EC (rank=2), PT N0—N12 (rank=3), erythrocytes/CC (4), thrombocytes/CC (5), stick neutrophils/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), eosinophils/CC (9), leucocytes/CC (10), monocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
Pharmacokinetic characterization of angiotensin IV analogs with therapeutic potential for cancer and dementia.
1. THESIS PROPOSAL DEFENSE ALENE MCCOY THESIS MENTOR: JOSEPH HARDING APRIL 30, 2009 “ Pharmacokinetic characterization of angiotensin IV analogs with therapeutic potential for cancer and dementia.”
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4. Is there a molecular target for both, age related dementia and cancer?
5. Angiotensins Binds to AT 1 & AT 2 : Blood pressure Vasoconstriction Binds to AT 4 : Cognition Memory
9. Effect of AT 4 Antagonists on Human Endothelial Cell Growth Control 10 -6 10 -8 10 -10 10 -12 0 50 100 150 PNB-0718 (M) Absorbance as Percent of Control HUVEC Cells Cell number estimated by MTT Mean +/- SEM, n=8
10. Effect of AT 4 Antagonists on Human Endothelial Cell Migration HUVEC Cells Cell Number Estimated by Visual Count Mean +/- SEM, n=8 control -8 -10 -12 -14 0 25 50 75 100 control -8 -10 -12 -14 Concentration (-log M) Average of Five Counts at High Power
11. Inhibition of Angiogenesis in the Mouse Aortic Ring Assay Control Treated Control PNB-0718 0.1 nM Angiogenesis (area) n=6 n=8 +/- SEM * p=0.012 *
12. Angiogenesis in Cancer Image obtained from roswellpark.org http://www.roswellpark.org/files/1_2_1/cancer_101/Lesson4/Lesson_4.pdf
13. Reduction of tumor growth. PNB-0718 Inhibits the Growth of Established Melanoma Tumors Time (Days) Tumor Volume (mm 3 ) Mean +/- SEM N=8 12 13 14 15 16 17 0 1000 2000 3000 Control PNB-0718 (2mg/kg/day) PNB-0718 (0.2 g/kg/day) = Injection
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17. Development of AT 4 Ligand Molecules Name Structure Activity Angiotensin IV Val-Tyr-Ile-His-Pro-Phe + PNB-0719 Nle -Tyr-Ile-His-Pro-Phe ++ PNB-0718 “ Norleual” Nle -Tyr- Leu- -His-Pro-Phe - - PNB-0405 d-Nle -Tyr-Ile- CH-CH-CH-CH-CH-CH-NH 2 OH - PNB-0408 Hexanoic acid -Tyr-Ile- CH-CH-CH-CH-CH-CH-NH 2 OH ++
18. Development of AT 4 Ligand Molecules Name Structure Activity Angiotensin IV Val-Tyr-Ile-His-Pro-Phe + PNB-0719 Nle -Tyr-Ile-His-Pro-Phe ++ PNB-0718 “ Norleual” Nle -Tyr- Leu- -His-Pro-Phe - - PNB-0405 d-Nle -Tyr-Ile- CH-CH-CH-CH-CH-CH-NH 2 OH - PNB-0408 Hexanoic acid -Tyr-Ile- CH-CH-CH-CH-CH-CH-NH 2 OH ++
24. Modeling of Physicochemical Properties of PNB-0718 Physicochemical Property Predicted Value Interpretation logP 0.68 slightly hydrophobic Fraction Unbound (to plasma proteins) 13.32 mostly bound to plasma proteins Effective Human Jejunal Permeability 0.28 not well absorbed in the gut (oral administration) Average Intestinal Permeability 0.08
40. Modeling of Physicochemical Properties Physicochemical Property Predicted Value Interpretation logP 1.45 hydrophobic Fraction Unbound (to plasma proteins) 42.68 ~50% bound to plasma proteins Effective Human Jejunal Permeability 1.53 moderately well absorbed in the gut (oral administration) Average Intestinal Permeability 0.39
58. Morris Water Maze Task of Spacial Memory Extra-maze cues hidden pedestal 4 - trials per day 120 seconds per trial 20 second on-pedestal rest between trials
59. Day 1 Day 2-3 Day 4 Morris Water Maze Representative Swim Patterns 1-2 Trials 3-4 Trials 13+ Trials 5-12 Trials
60. PNB-0408 reverses a chemically-induced learning deficit. Rat Performance in the Morris Water Maze
Ang I binds to nothing Ang II binds to AT1 And AT2 Ang III binds to same – CNS – short acting, probably more like neurotransmitter Ang IV binds to AT4 Ang IV not involved in renin-angiotensin system
Discovered trying to clone and purify AT1 -- Found a receptor that only binds Ang IV Binding sites widely distributed in terms of tissue, cell type and species. Evolutionarily conserved
Presence of AT 4 receptors on endothelial cells Importance of endothelial cells in regulating blood vessel growth (angiogenesis) *Numerous pathologies with aberrant blood vessel growth (too much or too little)
Presence on endothelial cells led to thinking about what role AT4 plays in endothelial cell activity Involvement in angiogenesis which is growth of new blood vessels Angiogenesis requires that endothelial cells proliferate and migrate Lab did series of studies on endothelial cell proliferation and migration Discuss figure Explain Mtt assay
Number of cells that have migrated across membrane? Stimulated by? Stained with cresyl violet
AT4 antagonists inhibit proliferation and migration – next step angiogenesis matrigel - an artificial extracellular matrix substitute Area estimated by measuring the distance between the farthest vessels. treat with norleual 10^-10M control receive vehicle measured area of angiogenic sprouting *point out effect is a lot more dramatic when comparing density vs area
In mice No injections prior to day 13 – rebound effect after injections stopped.
Gab-1 selective for c-Met.
Nle – improved affinity for receptor – why tried that? – hydrophobic branched chain hydrophobic straight chain What evidence? – binding studies (slide in spring semenar) Hexanoic acid – acetyl group: OH—CH—CH2—CH2—CH2—CH2—CH2– O Why 6AH? – increase hydrophobicity, stability– amide reduce ability of peptidase to cleave – why six? – approx same length as 3 Aas but w/o side chains
Not only do AT4 receptor antagonists inhibit endothelial cell growth…
“ Summary data” from western blots
+SA Murine breast cancer Elvax pellets
Before performing pharmacokinetic studies, it is helpful to determine physicochemical properties of molecule
Electrospray ionization
Vd suggests PNB-0718 resides predominantly in the central blood compartment.
Cell type?
MDCK Why not looking at 0413 instead? – We are but 0405 is prototype molecule.
Scattering – activity specific to c-Met activation Incubation: 4 days MDCK grown to confluency on round, glass cover slip
+SA Murine breast cancer Significant difference between control and treated group. Result doesn’t look quite as good as for 0718 but hoped to optimize dosing strategy after PK done.
Comparison of gut permeability to other orally administered drugs – intermediate.
Vd suggests PNB-0405 is extensively distributed outside the central blood compartment.
Summary data from western blots Potentiated HGF effect
One of the methods we use to measure the cognitive-enhance activity of our compounds is LTP. LTP believed to mimic molecular event underlying learning Help determine if our compounds are directly affecting hippocampal plasticity Hippocampus integrates and amplifies information then sends to neocortex for storage Stimulate CA3 in hippocampus – action potential runs down axons to dendrites of CA1 CA1 stimulation recorded
If interval between stimulations is far enough apart so that they don’t affect each other – baseline stimulation. Theta burst stimulation – thought to simulate a learning event – observed during active learning event. EPSP – excitatory post synaptic potential