Dan Salvail, Ph.D., Director of Pharmacology
His undergrad studies in Biochemistry were followed by graduate studies in Thoracic Physiology. Dan’s post-graduate work was in Cardiovascular Pharmacology and Cardiology.
“IPST, from its very beginning, has represented a hybrid; academic open-mindedness and flexibility paired with the dynamics of the private sector – where things move faster.”Email Dr Salvail
Dual antiplatelet and anticoagulant APAC prevents experimental ischemia–reperfusion-induced acute kidney injury
- Raimo Tuuminen Affiliated with Transplantation Laboratory Haartman Institute, University of HelsinkiDepartment of Cardiothoracic Surgery, Helsinki University Hospital
- Annukka Jouppila Affiliated with Helsinki University Hospital Research Institute
- Dan Salvail Affiliated with IPS Therapeutique
- Charles-E. Laurent Affiliated with IPS Therapeutique
- Marie-Claude Benoit Affiliated with IPS Therapeutique
- Simo Syrjälä Affiliated with Transplantation Laboratory Haartman Institute, University of HelsinkiDepartment of Cardiothoracic Surgery, Helsinki University Hospital
- Heikki Helin Affiliated with Division of Pathology, HUSLAB and Helsinki University Hospital
- Karl Lemström Affiliated with Transplantation Laboratory Haartman Institute, University of HelsinkiDepartment of Cardiothoracic Surgery, Helsinki University Hospital
The article – in its entirety – is available online at: http://link.springer.com/article/10.1007/s10157-016-1308-2
Renal ischemia–reperfusion predisposes to acute kidney injury (AKI) and mortality. APAC, mast cell heparin proteoglycan mimetic is a potent dual antiplatelet and anticoagulant inhibiting thrombosis in several vascular models.
Clinically relevant (0.06 and 0.13 mg/kg) and high (0.32 and 7.3 mg/kg) heparin doses of APAC and unfractionated heparin (UFH) were administered i.v. in pharmacological studies. Antithrombotic action of APAC and UFH was assessed with platelet aggregation to collagen, activated partial thromboplastin (APTT) and prothrombin (PT) times. Pharmacodynamics of [64Cu]-APAC or -UFH were monitored by PET/CT. Next, APAC and UFH doses (0.06 and 0.13 mg/kg) were i.v. administered 10 min prior to renal ischemia–reperfusion injury (IRI) in rats.
APAC in contrast to UFH inhibited platelet aggregation. During 0.06 and 0.13 mg/kg dose regimens APTT and PT remained at baseline, but at the high APTT prolonged fourfold to sixfold. Overall bio-distribution and clearance of APAC and UFH were similar. After bilateral 30-min renal artery clamping, creatinine, urea nitrogen and neutrophil gelatinase-associated lipocalin concentrations and histopathology indicated faster renal recovery by APAC (0.13 mg/kg). APAC, unlike UFH, prevented expression of innate immune ligand hyaluronan and tubulointerstitial injury marker Kim-1. Moreover, in severe bilateral 1-h renal artery clamping, APAC (0.13 mg/kg) prevented AKI, as demonstrated both by biomarkers and survival. Compatible with kidney protection APAC reduced the circulating levels of vascular destabilizing and pro-inflammatory angiopoietin-2 and syndecan-1. No tissue bleeding ensued.
APAC and UFH were similarly eliminated via kidneys and liver. In contrast to UFH, APAC (0.13 mg/kg) was reno-protective in moderate and even severe IRI by attenuating vascular injury and innate immune activation.
Ischemia–reperfusion injury Acute kidney injury Unfractionated heparin APAC
Abstract: Correlation between the thrombogenic Factor XIa and the thrombus score in the venous stasis rabbit model
Journal of Thrombosis and Haemostasis, Special Issue: Abstracts of the 62nd Annual Meeting of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis May 25–28, 2016, 14 (Suppl. 1) (2016) 1–168.
Background: Activated factor XI (FXIa) has been identiﬁed as the probable causative agent associated with thromboembolic events (TEE) following intravenous (IV) IgG (IVIG) product administration. Although there is no ofﬁcial method required by the regulator to validate the therapeutic safety of new batches of IVIG, the rabbit venous stasis assay, a prothrombogenic model, remains the model of choice for in vivo assessment of potential TEE.
Aims: This study aimed at standardizing the in vivo venous stasis rabbit model and quantifying the formed thrombi on a ranking system to evaluate the amount of FXIa in new batches of IVIG, in order to predict potential TEE. Methods: Anesthetized rabbits (2.5 to 3 kg) were infused with 10 ml/kg of 200 mM glycine, 5% BSA (Gycine-BSA) solution spiked with increasing doses of FXIa followed by a jugular venous stasis of 2-cm long for 15-min. The formed thrombus was scored according to a ranking system from 0 to 4.
Results: Spiked Glycine-BSA solution with 4 increasing doses of FXIa (0, 1, 1.5, 2 and 7 mU/mL) showed a dose-dependent clot formation response of 0, 0.5, 1.0, 2.33, and 3.75 respectively. The IVIG Gamunex and spiked Gamunex with 1.5 mU/mL of FXIa scored 0.8 and 3.5 respectively. Concomitantly, the bleeding time in rabbits treated with the spiked Glycine-BSA decreased from 220 s to 130 s with increasing doses of FXIa.
Conclusions: The current study demonstrates that the standardized venous stasis rabbit model is very sensitive to FXIa and that the thrombus score correlates with the amount of FXIa present in the test solution. Furthermore, this standardized model can be used to predict the potential for a TEE in batches of IVIG.
Poster: RP5063 Prevents Monocrotaline Induced Pulmonary Arterial Hypertension in Rats. Bhat L, Salvail D, Bouchard A, Hawkinson J, Cantillon M
Rationale: Pulmonary arterial hypertension (PAH) is characterized by constriction and remodeling of the pulmonary vasculature. Changes in serotonin synthesis, receptor activation and uptake via the serotonin transporter (SERT) have been reported in experimental and clinical PAH. Acute vascular constriction by chronic exposure to high levels of serotonin (5-HT) can be mediated by 5-HT1B and 5-HT2A receptors and thickening of cardiac valves through 5-HT2B receptor. RP5063 is a novel multimodal modulator of dopamine and serotonin receptors with high affinity for 5-HT2B (Ki=0.19 nM),5-HT2A (Ki=2.5 nM),5-HT1A, 5-HT6 and 5-HT7 receptors. In this study, the possibility that RP5063 could bind to these serotonin receptors to alleviate the symptoms of experimental PAH in rats was quantified by hemodynamic monitoring and histological examination.Methods: Adult male Sprague-Dawley rats were given a single s.c. injection of 50 mg/kg monocrotaline on Day 0. The animals were monitored for 28 days while PAH symptoms progressed. 10 animals per group received either vehicle, 1 mg/kg, 3 mg/kg, or 10 mg/kg RP5063, b.i.d. or 50 mg/kg sildenafil, b.i.d. by oral gavage for 28 days. Functional observations and body weight were monitored and blood samples were taken weekly. On Day 29, terminal surgery involving intra-arterial catheters (femoral and pulmonary), right ventricle, ECG recording and SO2 monitoring was performed to acquire a complete hemodynamic profile of the animal’s status. The lungs and heart were harvested, fixed, and embedded before being stained (H&E) and submitted to morphometric analysis to define the extent of the remodeling.Results: Monocrotaline increased pulmonary diastolic and systolic pressures from 11.4 to 21.1 and 26.7 to 64.0 mmHg, resp. 10 mg/kg RP5063 significantly lowered the pulmonary diastolic and systolic pressures to 14.7 and 43.0 mmHg, resp. 10 mg/kg RP5063 reduced right-ventricle remodeling (Fulton Index decreased from 0.47 to 0.32) and lowered respiratory resistance from 20.9 to 10.7 mmHg/mL/sec, suggesting that RP5063 decreased pulmonary edema and fibrosis. Histology revealed a dose-dependent decrease of small vessel wall thickness from 41.9 to 25.3% (10 mg/kg RP5063) of the vessel diameter. Vascular remodeling produced 33.7% less muscular vessels in 10 mg/kg RP5063 -treated animals with significantly less alveolar macrophage infiltration in treated animals.
Conclusions: Combined with the morphometric evidence of limited lung remodeling, these results support a greater functional and structural benefit from RP5063 compared to a 5-fold higher dose of sildenafil, a current off-label treatment for clinical PAH.
Article: Therapeutic antibody-induced vascular toxicity due to off-target activation of nitric oxide in cynomolgus monkeys. Pai R1, Ma N1, Connor AV1, Danilenko DM1, Tarrant JM1, Salvail D2, Wong L1, Hartley DP1, Misner D1, Stefanich E3, Wu Y4, Chen Y4, Wang H5, Dambach DM1. Toxicol Sci. 2016 Feb 25. pii: kfw037. [Epub ahead of print]
- 1* Safety Assessment, Genentech Inc., 1 DNA Way, South San Francisco, California 94080.
- 2IPS Therapeutique, Sherbrooke, Quebec, Canada.
- 3Preclinical & Translational Pharmacokinetics, Genentech Inc., 1 DNA Way, South San Francisco, California 94080.
- 4Antibody Engineering, Genentech Inc., 1 DNA Way, South San Francisco, California 94080.
- 5* Safety Assessment, Genentech Inc., 1 DNA Way, South San Francisco, California 94080 firstname.lastname@example.org.
Abstract: PRO304186, a humanized monoclonal antibody targeting soluble interleukin-17 A and F, was developed for autoimmune and inflammatory disease indications. When administered to cynomolgus monkeys PRO304186 induced unexpected adverse effects characterized by clinical signs of hematemesis, hematochezia and moribundity. Pathology findings included hemorrhage throughout the gastrointestinal tract without any evidence of vascular wall damage or inflammatory cellular infiltration. Mechanistic investigation of these effects revealed mild elevations of serum MCP-1 and IL-12/23 but without a classical proinflammatory profile in PRO304186-treated animals. In vitro studies demonstrated off-target effects on vascular endothelial cells including activation of nitric oxide synthase leading to production of nitric oxide (NO) accompanied by increased mitochondrial membrane depolarization, glutathione depletion and increased paracellular permeability. Additionally, endothelial cell-PRO304186-conditioned medium reduced myosin light chain phosphorylation in vascular smooth muscle cells. Furthermore, an ex vivo study utilizing segments from cynomolgus aorta and femoral artery confirmed PRO304186-induced endothelium-dependent smooth muscle relaxation and vasodilation mediated via NO. Finally, a single dose of PRO304186 in cynomolgus monkeys induced a rapid and pronounced increase in NO in the portal circulation that preceded a milder elevation of NO in the systemic circulation and corresponded temporally with systemic hypotension; findings consistent with NO-mediated vasodilation leading to hypotension. These changes were associated with non-inflammatory, localized hemorrhage in the gastrointestinal tract consistent with hemodynamic vascular injury associated with intense local vasodilation. Together, these data demonstrate that PRO304186-associated toxicity in monkeys was due to an off-target effect on endothelium that involved regional NO release resulting in severe systemic vasodilation, hypotension and hemorrhage.
© The Author 2016. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: email@example.com.
KEYWORDS: hemodynamic vascular injury; nitric oxide; non-human primates; off-target toxicity; splanchnic circulation; therapeutic antibody
Poster: LPS-induced Cytokine Storm Produces QTc Prolongation Which Can be Prevented by an Antiinflammatory Eutectic Blend. Annie Bouchard1, Peter Sordillo2, Lawrence Helson2, George Shopp3, Sabrina Baillargeon1, Walter Shaw4, Dany Salvail1
1IPS Therapeutique Inc., Sherbrooke, QC, Canada, 2SignPath Pharma Inc., Quakertown, PA, USA,
3Shopp NonClinical Consulting LLC., Boulder, CO, USA, 4Avanti Polar Lipids Inc., Alabaster, AL, USA
Abstract: There is increasing evidence that excess levels of pro-inflammatory cytokines play a major role in the pathogenesis of the prolonged QT syndrome. Inversely, blockers such as tocilizumab (IL-6), or anticytokine antibodies (TNFα) contribute to a shortening of the previously-prolonged QT interval. In this study, LPS and Kdo2-Lipid-A were used to induce cytokine release in guinea-pigs with concomitant ECG monitoring and blood draws, followed by Q-ELISA measurement of cytokine production. The guinea pig was selected because it yields reliable QTc prolongation as a result of pro-arrhythmic challenge, with consistently visible T-waves on the ECG. Male adult guinea pigs received 300 μg/kg LPS at time 0, and had ECGs analyzed at 1h, 2h, and 4 hours post-LPS, with simultaneous blood draw. Animals receiving LPS only exhibited a 8-msec increase in QTc after 1h post-LPS, when TNFα levels were maximal at 5.5-fold the pre-LPS values. A 29-msec QTc prolongation 2h post-LPS correlated with 7- and 9- fold increases in IL-1β and IL-6, respectively. The QTc prolongation remained (27 msec) after 4 hours post-LPS, when the animals were euthanized. When 9 mg/kg EU8120 (a lipid blend shown to prevent IKrchannel block by a variety of hERG blockers) was given 1 hour prior to LPS-induction, QTc prolongation was limited to 5 ms after 2 hours, and completely prevented at 1 and 4 hours post-LPS. Plasma levels of TNFα, IL1β, and IL-6 were significantly lower in EU8120-administered animals. It is suggested that EU8120 reduces QT prolongation through suppression of pro-inflammatory cytokines. Presented at Safety Pharmacology Society (SPS) Annual Meeting in Prague on Tuesday, September 29th.