Suven Life Sciences Limited · NSE:SUVEN
Suven Life Sciences — Technical Presentation (2016)
127 slides from Suven Life Sciences's technical presentation, 2016
Slide 1: Drug Discovery Collaboration
Drug Discovery Collaboration Discovery Research Suven Life Sciences Serene Chambers, Road-5, Avenue-7, Banjara Hills, Hyderabad-500034, India. Contacts: [email protected], [email protected]
Slide 2: Vision and Mission
Drug Discovery Collaboration Vision Search for new CNS therapies Become the leading company focused on the treatments for unmet medical need in Mental Health Mission Health for Patients and Value for Partners Excellent Pipeline of CNS drugs to improve Cognition, treat Depression and Pain Novel Mecha
Slide 3: Index
Index Slides Slides • Suven Company Overview 04 - 05 • In-vitro Biology 32 – 59 (Assay Development, Validation, Screening, Profiling) • Suven Discovery Profile 06 • In-vitro ADME & In-vivo PK 60 - 74 • Integrated Drug Discovery 08 - 09 • Target engagement : Microdialysis, 75 – 89 Receptor Occupancy
Slide 4: Suven Life Sciences: 27 years of Pharmaceutical Leadership
Suven Life Sciences 27 years of Pharmaceutical Leadership 2005 Drug Discovery & Development 1989 Support Services (DDDSS) Incorporated by Mr. Venkat Jasti Major Pharma & Biotech (USA, Europe) 1995 IPO, Listed on NSE and BSE in India 2006 Collaborative Research Program Eli Lilly, USA 1989 Contract Re
Slide 5: Suven Life Sciences - Business Model
Suven Life Sciences - Business Model Drug Discovery Formulation Development Clinical Supplies Lead Process Custom Hit Lead Pre Clinical Clinical Trials Analytical & Manufacturing & Optimization Research Synthesis Regulatory Packaging Services Drug Discovery & Development Contract Research And Suppor
Slide 6: Suven Discovery - Profile
Suven Discovery - Profile More than 14 years of Discovery and Development experience Worked on 8-10 Central Nervous System targets in drug discovery First discovery project delivered SUVN-502 (5-HT6 antagonist for Alzheimer's Disease). Phase II POC for Alzheimer's Disease in aged population is curre
Slide 7: Drug Discovery and Developmental Research
Drug Discovery and Developmental Research Company Confidential 7 Copyright © 2016 Suven Life Sciences Limited
Slide 8: Suven Integrated Drug Discovery
Suven Integrated Drug Discovery Company Confidential 8 Copyright © 2016 Suven Life Sciences Limited
Slide 9: Suven Discovery Research
Suven Discovery Research Medicinal / Organic / Scale-up Chemistry Analytical Chemistry Physicochemical Properties and Biopharmaceutical characterization Assay development and validation In-vitro Screening; Functional Assays In-vitro ADME Assays In-vivo Pharmacokinetics In-vitro, In-vivo Metabolism I
Slide 10: Suven Discovery Pipeline
Suven Discovery Pipeline Neurodegenerative Diseases l III ery La cted II ca I e- e- ch e- i MCI / Cognition / Alzheimer's / IND ov lin as as un pe as sc ec Schizophrenia Ph Ph Ex Ph Di Pr SUVN-501 (5-HT 6 Antagonist) SUVN-502 (5-HT 6 Antagonist) 2015 2020 SUVN-507 (5-HT 6 Antagonist) SUVN-512 (5-HT
Slide 11: Suven Discovery Pipeline
Suven Discovery Pipeline al III ery II La cted I ic Pain / Major Depressive Disorder e- e- ch e- lin D ov as (MDD) / Psyciatric Discorders as pe un IN as eC sc Ph Ph Ex Ph Di Pr nAChR Alpha-4 Beta-2 Antagonist (Pain) SUVN-911 (nAChR Alpha-4 Beta-2) (MDD) 2016 2016 CB2 Agonist (Pain) Multi Modal (Psy
Slide 12: Suven Patent Pipeline
Suven Patent Pipeline PCT and National l 02 03 04 05 06 07 08 09 10 11 12 13 14 15 16 ta Phase To 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 Discovery Research Applications 1 8 4 3 2 3 3 4 4 1 2 1 5 _ 2 43 Published _ 3 7 3 1 5 3 2 2 5 2 1 2 2 3 41 National Phase _ _ 3 3 1 1 2 3 1 2 5 3 1 1 2 28 e
Slide 13: Suven Patent Pipeline
Suven Patent Pipeline d e an ia or pe PO el n al al Patents Granted A ap pa ra r ro US Ze I st AR ng Ja Is Eu Au w Si Ne Discovery (Product Patents) 21 24 23 18 23 24 9 2 Process Patents 2 5 2 2 _ 1 1 _ a a ric nk ico u zil ea a Af ca di La a x r In Ma Ko Br Me h i ut Sr So Discovery (Product Patent
Slide 14: Suven Published PCT Patent Applications Discovery Research
Suven Published PCT Patent Applications Discovery Research Year Publication Numbers Number of Publications 2016 WO2016027275, WO2016027276, WO2016027277 3 2015 WO2015083179, WO2015092804 2 2014 WO 2014/030170, WO 2014/147636 2 2013 WO 2013/042135 1 2012 WO 2012/029070, WO 2012/114348 2 WO 2011/03034
Slide 15: Suven Published PCT Patent Applications Process Research
Suven Published PCT Patent Applications Process Research Number of Year Publication Numbers Publications WO 2015/008294, WO2015037013, WO/2015/155664, 2015 5 WO/2015/162506, WO/2016/027277 2010 WO 2010/061398 1 2009 WO 2009/007995, WO 2009/007998 2 2007 WO 2007/026375, WO 2007/020654, WO 2007/094007
Slide 16: Suven Discovery Scientists Qualifications / Experience
Suven Discovery Scientists Qualifications / Experience Scientists Breakdown Average Years MS / Average Years Technician / Ph.D. Experience M.Pharm Experience Lab Support Chemistry 10 6 - 9 Years 60 2 - 6 years 12 Biology 12 >10 Years 95 2 - 8 years 25 DMPK 7 >10 Years 45 2 - 5 years 12 Management 2
Slide 17: Suven Discovery Scientists
Suven Discovery Scientists Most of the Scientists regularly attend International conferences like ACS, AAIC, SFN, AAPS, CRS, SOT, In-vivo and ISSX Most of the Scientists from Assay development, Assay Validation, ADME, Microdialysis, Receptor Occupancy, PK, Toxicology had spent 2-3 weeks in training
Slide 18: Suven Discovery Process: Hit-to-Candidate Selection
Suven Discovery Process: Hit-to-Candidate Selection Hit to Lead = 9 to 14 months Primary screening, ADME Rodent Rat Target Primary In-vivo Binding and/or Surrogate Bioavailability Tissue Efficacy Assay Functional Assays1 2w, 25 mg Distribution 1w, 200 mg Stage 1 (rat & human) 1w, 5 mg 1w, 30 mg 1w,
Slide 19: Results/Data - Communication
Results/Data - Communication Collaborative Projects Data upload in dedicated eRoom on real time Secured Tunnel for safe email exchanges Weekly WebEx for Chemistry, Biology and project progression Regular teleconference with the collaborative partners Access to structures is restricted to g
Slide 20: Medicinal Chemistry
Suven Discovery Research Medicinal Chemistry
Slide 21: Discovery Research - Medicinal Chemistry
Discovery Research - Medicinal Chemistry Core Capabilities Hit Generation and Hit follow up Hit to lead generation with consideration of potential IP issues Lead Optimization and selection of clinical development candidate Scale-up Chemistry Support Identification, synthesis and characteri
Slide 22: Discovery Research - Medicinal Chemistry
Discovery Research - Medicinal Chemistry Collaborative Capabilities Hit Generation and Hit follow up Independent or Customer supported Hit to lead and Lead generation Independent or Synergistic work for Lead Optimization with the Collaborator Scale-up Chemistry Support for developmental prog
Slide 23: Case study -1: Internal Discovery Project - Histamine H3 antagonists
Case study -1: Internal Discovery Project - Histamine H3 antagonists Optimization of ADME properties O R1 hH3 Ki < 5 to 10 nM, Initial hits N Poor brain penetration N H Longer half life Poor oral exposure Confirmational constrained to improve ADME properties O R1 hH3 Ki < 5 to 10 nM, Initi
Slide 24: Discovery Research - Medicinal Chemistry
Discovery Research - Medicinal Chemistry The People - Core Strength Well qualified, Trained and Committed Scientific Staff Well versed with Medicinal & Contemporary Chemistry Skilled in various synthetic techniques Highly interactive with biologists Upto-date knowledge of IP issues, highes
Slide 25: The Facilities - Labs
Discovery Research - Medicinal Chemistry The Facilities - Labs Spacious and well equipped air conditioned laboratories Spacious 8 fume-hoods per lab Integrated write up area and IP secure areas within the lab for scientists. State of the art purification systems like Combiflash Non-convent
Slide 26: The Facilities - Chiral Chemistry Capabilities
Discovery Research - Medicinal Chemistry The Facilities - Chiral Chemistry Capabilities Expertise in designing asymmetric synthesis Expertise in traditional chiral resolutions via diastereoisomeric salt preparation and crystallizations Analytical chiral columns for chiral analysis Method dev
Slide 27: The Facilities - Literature
Discovery Research - Medicinal Chemistry The Facilities - Literature Online access to major Medicinal and Organic chemistry journals Access to STN and Scifinder In-house library Access to printed literature and databases Company Confidential 27 Copyright © 2016 Suven Life Sciences Limited
Slide 28: Analytical Support Facilities
Discovery Research - Medicinal Chemistry Analytical Support Facilities Analytical Testing (API, Impurities, Intermediates, Starting Materials, Drug Product) Stability studies (API and Drug Product) Analytical Method Development & Validation (Chemical, Chromatographic and Chiral) (Starting Mate
Slide 29: Analytical Support Facilities
Discovery Research - Medicinal Chemistry Analytical Support Facilities 400 MHz NMR (Bruker) with three different probes – dual, multinuclear and fluorine Dedicated mass spectrometer for med chem - LC MS/MS API-2000 (Applied biosystems) Five Agilent HPLC systems with UV, PDA, Fluorescence and R
Slide 30: Chemistry, Manufacturing and Control (CMC)
Chemistry, Manufacturing and Control (CMC) • Good expertise in the synthesis and scale up of the APIs, intermediates, fine chemicals, impurities and metabolites • Excellent laboratory facilities for scale up, good manufacturing facilities, inclusive of FDA approved production facilities • Expertise
Slide 31: Chemistry, Manufacturing and Control (CMC)
Chemistry, Manufacturing and Control (CMC) • Control of the impurities through diligent planning and route selection. Identification of the formation of solvates, hydrates etc • Expertise in crystallization studies, polymorph identification and control • Salt screen and selection based on solubility
Slide 32: In-vitro Biology
Suven Discovery Research In-vitro Biology
Slide 33: In-vitro Biology
In-vitro Biology A group of dedicated scientists with overall expertise in: Biochemistry, Molecular Biology and Cell Biology Encompassing drug target class: GPCRs, Nuclear Receptors, Monoamine transporters, ion channels and enzymes Expertise: Assay Development and Validation, Screening and Profiling
Slide 34: Biochemistry and Molecular Biology
Biochemistry and Molecular Biology Human cDNA Synthesis and Cloning Expression of Recombinant Proteins in E. coli, Baculovirus and Mammalian Cells Purification and Characterization of Recombinant Proteins Establishment, identification and characterization of stable cell lines (both CHO and H
Slide 35: Target Class GPCRs
Target Class GPCRs Radioligand binding assays cAMP Measurement and Reporter Gene Driven Assays Functional Cell-based Assays (Signal Transduction) Company Confidential 35 Copyright © 2016 Suven Life Sciences Limited
Slide 36: Target Class Ion Channels
Target Class Ion Channels Radioligand binding assays Patch clamp assay using automated electrophysiology instrument Patchliner from Nanion, for ligand gated ion channel: P2X7 as well as voltage gated ion channels: Sodium Channels - Nav 1.4, 1.5, 1.6, 1.7 and Potassium Channels - Kv 10.1 and Kv11
Slide 37: Target Class Monoamine Transporters
Target Class Monoamine Transporters Radioligand binding assays Functional uptake assays Nuclear Receptors Reporter gene based assays Enzyme-based assays Historical expertise in establishment and validation of assays and screening of compounds using kinase, peptidase and phosphatase as drug t
Slide 38: Protein - Protein Interactions and Other Assays
Protein - Protein Interactions and Other Assays Yeast Two-Hybrid System and other yeast based assays His Tag Pull Down Immunohistochemistry CYP Induction: PXR transactivation assay In-vitro Cytotoxicity and Phospholipidosis assay Company Confidential 38 Copyright © 2016 Suven Life Sciences
Slide 39: Assay Validation TRT
Assay Validation TRT Potency Spreadsheet . . . . Assay Run 1 & Run 2 . . . . Assay Outside Potency Ratio vs Geometric Mean Potency Compound Run 1 Run 2 Ratio Limits 10 3.5 2.5 1.400 Ratio 83.74 101.2 0.827 Page Limit Boundaries 26.66 30.5 0.874 MR 89.5 103 0.869 RLs 18.6 16.7 1.114 Ratio 95.35 141.3
Slide 40: Assay Validation PUT
Assay Validation PUT Day 2, Plate 1 row 1 2 3 4 5 6 7 8 9 10 11 A 2362 1637 142 2384 1580 145 2388 1601 147 2300 1612 B 2369 1601 131 2328 1615 156 2387 1625 151 2380 1663 C 2391 1580 132 2338 1582 140 2392 1630 156 2338 1658 D 2360 1636 138 2367 1614 145 2358 1674 141 2376 1613 E 2310 1609 142 2309
Slide 41: Assay Validation PUT
Assay Validation PUT Day 1 Plate 1 Day 1 Plate 1 3000 3000 2500 2500 2000 Response Max 2000 Max 1500 Mid Mid 1500 1000 Min Min 1000 500 500 0 0 0 12 24 36 48 60 72 84 96 0 8 16 24 32 40 48 56 64 72 80 88 96 Well - Row Orie nted Well - Colum n Oriented Day 1 Plate 2 Day 1 Plate 2 3000 3000 2500 2500
Slide 42: Assay Validation PUT
Assay Validation PUT Ratio EC50/IC50/Ki within Days (larger over smaller) Plate 1 - Plate 1 - Plate 2 - Plate 2 Plate 3 Plate 3 Day 1 1.1 1.0 1.0 Meets Criterion Day 2 1.1 1.1 1.0 Meets Criterion Day 3 1.0 1.1 1.0 Meets Criterion Ratio EC50/IC50/Ki between Days (larger over smaller) Day 1 - Day 2 1.
Slide 43: A reporter gene driven cell based assay
A reporter gene driven cell based assay Company Confidential 43 Copyright © 2016 Suven Life Sciences Limited
Slide 44: Cell Based Assay Validation
Cell Based Assay Validation 100 Full Agonist 75 Antagonist Partial Agonist % Activity 50 Inverse Agonist Inac tive C ompound 25 0 -25 -1 0 1 2 3 4 Log[Compound]nM Compounds representing various class and varying affinities are selected for an assay validation EC50, IC50, and Kb values are derive
Slide 45: Patch Clamp Assay
Patch Clamp Assay Patchliner with 8 Channel Ion Channel Company Confidential 45 Copyright © 2016 Suven Life Sciences Limited
Slide 46: Voltage gated ion channel: Potassium – Kv 11.1 (hERG)
Voltage gated ion channel: Potassium – Kv 11.1 (hERG) Cell line : Stably expressing recombinant HEK293 hERG cell line Test item: Astemizole, Haloperidol and Quinidine Cells with membrane resistance of > 1 GOhms; Currents of > 200 pA; without rundown of current were taken into consideration for
Slide 47: Voltage gated ion channel: Potassium – Kv 11.1 (hERG)
Voltage gated ion channel: Potassium – Kv 11.1 (hERG) 1.0 Astemizole 'Control' 'Astemizole 10 nM' 200 'Astemizole 100 nM' 0.8 'Astemizole 300 nM' 'Astemizole 1 uM' 'Astemizole 10 uM' Astemizole 100 0.6 pA 0.4 0 0.2 -100 0.0 0.0 0.5 1.0 1.5 100 pM 1 nM 10 nM 100 nM 1 µM 10 µM s 1.0 Haloperidol 'Contr
Slide 48: Voltage gated ion channel: Potassium – Kv 11.1 (hERG)
Voltage gated ion channel: Potassium – Kv 11.1 (hERG) 1.0 Qunidine 'Control' 600 'Quinidine 300 nM' 'Quinidine 1 uM' 0.8 'Quinidine 3 uM' 'Quinidine 10 uM' 400 0.6 200 Quinidine pA 0.4 0 -200 0.2 -400 0.0 10 nM 100 nM 1 µM 10 µM 0.0 0.5 1.0 1.5 s Obtained IC50 value Reported IC50 value S.No Compound
Slide 49: Voltage gated ion channel: Sodium – Nav 1.5
Voltage gated ion channel: Sodium – Nav 1.5 Cell line : Stably expressing recombinant HEK293 Nav 1.5 cell line Test item: Tetracaine, Lidocaine, Proparacaine and Tetrodotoxin Cells with membrane resistance of > 1 GOhms; Currents of > 2 nA; without rundown of current were taken into considerati
Slide 50: Voltage gated ion channel: Sodium – Nav 1.5
Voltage gated ion channel: Sodium – Nav 1.5 1.0 Tetracaine 0 0.8 -1 Control Tetracaine 100 nM Tetracaine -2 Tetracaine 300 nM Tetracaine 1 uM 0.6 Tetracaine 3 uM nA Tetracaine 10 uM -3 Tetracaine 30 uM Tetracaine 100 uM 0.4 -4 0.2 -5 0.0 0 10 20 30 40 1 nM 10 nM 100 nM 1 µM 10 µM 100 µM ms 1.0 Lidoc
Slide 51: Voltage gated ion channel: Sodium – Nav 1.5
Voltage gated ion channel: Sodium – Nav 1.5 Proparacaine_HCl 0 0.8 -1 Control -2 Proparacaine HCl 100 nM Proparacaine Proparacaine HCl 300 nM Proparacaine HCl 1 uM 0.6 -3 Proparacaine HCl 3 uM nA Proparacaine HCl 10 uM Proparacaine HCl 30 uM 0.4 -4 Proparacaine HCl 100 uM -5 0.2 -6 0.0 0 10 20 30 40
Slide 52: Voltage gated ion channel: Sodium – Nav 1.5
Voltage gated ion channel: Sodium – Nav 1.5 Obtained IC50 value Reported IC50 value S.No Compound (µM) (µM) 1 Tetracaine 1.3 1.7 2 Lidocaine 258 260 3 Proparacaine HCl 1.8 1.9 4 Tetrodotoxin 5.3 4.3 Company Confidential 52 Copyright © 2016 Suven Life Sciences Limited
Slide 53: Ligand gated ion channel: ATP – Purinergic P2X7
Ligand gated ion channel: ATP – Purinergic P2X7 Cell line : Stably expressing recombinant HEK293 P2X7 cell line Test item: Agonist – ATP and Bz ATP, Antagonist - Suramin Cells with membrane resistance of > 1 GOhms; Currents of > 1 nA; without rundown of current were taken into consideration fo
Slide 54: Ligand gated ion channel: ATP – Purinergic P2X7
Ligand gated ion channel: ATP – Purinergic P2X7 1.0 ATP 0.0 0.8 ATP 250 uM -0.5 ATP ATP 500 uM ATP 1 m M 0.6 nA ATP 2 m M ATP 3 m M 0.4 -1.0 0.2 -1.5 0.0 0 1 2 3 4 5 3 4 5 6 7 8 9 2 3 0.001 s 1.0 M 0 Bz_ATP -1 0.8 BZ AT P 100 uM -2 BZ ATP 250 uM BZ ATP 500 uM 0.6 Bz ATP BZ ATP 750 uM nA BZ ATP 1 mM
Slide 55: Ligand gated ion channel: ATP – Purinergic P2X7
Ligand gated ion channel: ATP – Purinergic P2X7 1.0 Suramin -0.5 0.8 -1.0 Bz ATP 1mM 0.6 Suramin 10 uM + Bz ATP 1mM Suramin nA Suramin 100 uM + Bz ATP 1mM Suramin 1 mM + Bz ATP 1mM -1.5 Suramin 10 mM + Bz ATP 1m M 0.4 0.2 -2.0 0.0 0 1 2 3 4 5 100 nM 1 µM 10 µM 100 µM 1 mM 10 mM s S.No Compound Obtai
Slide 56: Cytotoxicity assessment
Cyototoxicity assessment MTT Assay 100 M 100 10 M Test System: HepG2 (Human hepatocellular Carcinoma), 75 1 M HEK293 (Human embryonic kidney) and % Viability 50 IMR32 (Human neuroblastoma) 25 Test item: 0 Tamoxifen (Positive control) Tamoxifen Acetaminophen Compound-1 Acetaminophen 100 Test
Slide 57: Phospholipidosis
Phospholipidosis Test System: CHO-K1 (Chinese Hamster Ovary cell line) and CHL/IU (Chinese Hamster Lung) Test item: Amiadarone (Positive control) Acetaminophen (Negative Control) Fluoxetine Test Conc.: 1 µM - 100 µM Incubation: 24 hour Assessment: Fluorescent Microscopy Fluorometry (Fluore
Slide 58: Phospholipidosis
Phospholipidosis Vehicle Fluoxetine (30 µM) Acetamoinophen (100 µM) (Positive Control) (Negative Control) Phospholipidosis Fluorometric Assay AMIODARONE 120 ACETAMINOPHEN PLD Inducing potential (%) 100 FLUOXETINE 80 60 40 20 0 30 M 20 M 15 M 10 M 5 M 3 M Company Confidential 58 Copyright ©
Slide 59: List of In-vitro Assays
List of In-vitro Assays In Vitro Assay Inventory at Suven Target Assay Type Target Assay Type GPCRs Transporters Adenosine A2A Reporter Gene Driven Cell Based SERT Radioligand binding/ Functional Uptake Adrenoceptor alpha 1B Radioligand binding/ Reporter Gene Driven Cell Based DOPT Radioligand bindi
Slide 60: In-vitro ADME, In-vivo PK, Regulatory Bioanalysis
Suven Discovery Research In-vitro ADME In-vivo PK Regulatory Bioanalysis
Slide 61: In-vitro ADME
In-vitro ADME • Solubility • Lipophilicity • Permeability assay (PAMPA & CACO-2) • Metabolic stability assay (Hepatic, intestinal microsomes and hepatocytes) • CYP Inhibition assay – 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 & 3A4 (Supersomes and microsomes) • Time Dependent Inhibition assay (Human liver m
Slide 62: In-vivo PK
In-vivo PK In Vivo Dosing In Vivo Studies − IV bolus (single and cassette) − Bioavailability − IV infusion − Bioequivalence − Oral − PK in target site (CNS or peripheral) − Sucutaneous − Single and multiple-dose pharmacokinetics − Intra-nasal − Tissue distribution (cold and 14C labelled) − Sublingua
Slide 63: Surgical Models
Surgical Models • Jugular vein Cannulated Rats, Guinea pigs & Hamsters • Femoral vein Cannulated Rats & Guinea pigs • Portal vein Cannulated Rats and Guinea pigs • Carotid artery Cannulated Rats • Abdominal aorta Cannulated Rats • Bile Duct Cannulated Rats • Duodenal, Jejunum, Ileum and Colon Cannul
Slide 64: Bi-directional Permeability assay
Bi-directional Permeability assay 7.5 Efflux Ratio Test System: Caco-2 Efflux Ratio 5.0 Test item: Digoxin (Standard) 2.5 Threshold and test compounds 0.0 Digoxin TC #1 TC #2 TC #3 TC #4 Test Conc.: 10 µM 150 Permeability 125 Incubation: 1 hour 100 Papp (10-6 cm/sec) 75 Membrane TEER and L
Slide 65: P-glycoprotein (P-gp) Substrate assessment
P-glycoprotein (P-gp) Substrate assessment Test System: Caco-2 Digoxin assessed on ten different days Test item: Digoxin 30 A-B (P-gp Substrate) B-A Papp (x 10 -6 cm/sec) 20 Test Conc.: 10 µM 10 0 Days 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 10 16 Assessment: - / + Verapamil - Verapamil + Ver
Slide 66: P-glycoprotein (P-gp) Inhibitor assessment
P-glycoprotein (P-gp) Inhibitor assessment 35 D ig oxin P er meability 20 30 Papp (10-6 cm/sec) Test System: Caco-2 Digoxin Efflux Ratio 25 Digoxin D ig o xin E fflu x R atio 20 15 15 10 Test item: Verapamil 5 0 10 A- B B- A A- B A- B B- A B- A (Positive control) Phenelzine 5 D igoxin 10 M Ver
Slide 67: CYP3A Time Dependent Inhibition (Single point screen)
CYP3A Time Dependent Inhibition (Single point screen) Matrix : Pooled Human Liver Microsomes (Mixed Gender) Drug Normal High activity CYP3A CYP3A Test Concentration : range spanning 10 fold to IC50 µM Indinavir 0 0 Protein concentration : 0.5 mg/ mL 0.25 mg/mL Ketoconazole 0 0 Incubation time : 10 &
Slide 68: CYP3A Time Dependent Inhibition (IC50 shift)
CYP3A Time Dependent Inhibition (IC50 shift) Matrix : Pooled Human Liver Microsomes (Mixed Gender) Test Concentration : range spanning 10 fold to IC50 µM Protein concentration : 1.0 mg/ mL 0.1 mg/mL Incubation time : 10 & 30 min ± NADPH Substrate : Midazolam Incubation time : 5 min % Activity (+ NAD
Slide 69: CYP3A4: Time dependent Inhibition (Inactivation kinetics)
CYP3A4: Time dependent Inhibition (Inactivation kinetics) Preincubation assay- 5.00 Test compound: Troleandomycin No com pound y = -0.009x + 4.61 Protein concentration: 0.5 mg/mL 4.00 R2 = 0.6928 Test Compound Concentration: 0, 0.312, 0.625, 1.25, 2.5, 5, 10 µM 0.312 µM 3.00 y = -0.0497x + 4.61 Prei
Slide 70: CYP Induction: Fold Change in Gene Expression
CYP Induction: Fold Change in Gene Expression of CYP1A2, CYP2B6 & CYP3A4 in Three Donors 84 Omeprazole (CYP1A2 inducer) 72 Phenobarbital (CYP2B6 inducer) Rifampicin (CYP3A4 inducer) 60 Fold change relative to 50.5 Control 48 44.6 36 33.8 24 10.8 10.7 12 7.5 6.9 5.9 6.7 0 HH1032 HH1007 HH1031 4 fold
Slide 71: In-vitro Reactive Met ID evaluation (Ticlopidine)
In-vitro Reactive Met ID evaluation (Ticlopidine) Compound : Ticlopidine Matrix : SD Rat liver microsomes Instrumentation and Analytical Method LC-MS/MS : API-4000 Qtrap (Applied Biosystems) Ion source : Turbospray HPLC : Shimadzu SIL HTc Column : Atlantis dC18, 4.6 x 150 mm, 5 µ Mobile phase : 0.1
Slide 72: In-vitro Reactive Met ID evaluation (Ticlopidine)
In-vitro Reactive Met ID evaluation (Ticlopidine) Cl S N Ticlopidine O Epoxidation S-oxidation Cl S Cl S O N N GSH GSH Cl S OH GS N Summed ion chromatogram of Parent, M1 and M2 •Both M1 and M2 were identified at 100µM of substrate. Proposed structure of Ticlopidine-GSH conjugate P: m/z 264 (Parent);
Slide 73: Dog PK: Assessment of pH and food effect on absorption
Dog PK: Assessment of pH and food effect on absorption of Test Compound Effect of Food on the Absorption of Test Compounds Effect of Food on the Absorption of Test Compounds Test compound (as suspension in capsule) administered by oral Test compound administer by oral gavage in male Beagle dogs at g
Slide 74: Regulatory Bioanalysis and Formulation Analysis
Regulatory Bioanalysis and Formulation Analysis Exclusive advanced analytical laboratory accredited with ISO:IEC 17025 for quality systems since 2005 Developed more than 150 sensitive and challenging bioanalytical methods Performed bioanalysis for Phase-1 first in human studies and submitted dossier
Slide 75: Target Engagement: Receptor Occupancy, Microdialysis, EEG
Suven Discovery Research Target Engagement: Receptor Occupancy Microdialysis Electroencephalography in Rodents (EEG)
Slide 76: LC-MS/MS (or) LSA Based Receptor Occupancy Methodology
LC-MS/MS (or) LSA Based Receptor Occupancy Methodology Formulation Test compound dosing p.o./s.c./ i.p./i.v. Animal model: rat/ mouse/ guinea pig LC-MS/MS based Liquid scintillation analyzer (LSA) based Non-radiolabelled tracer (i.v.) dose at Tmax of test Radiolabelled ([3H]) tracer (i.v.) dose at T
Slide 77: List of Validated Receptor Occupancy Assays
List of Validated Receptor Occupancy Assays Receptor Tracer Specific region Receptor Tracer Specific region #, * 5-HT1A WAY-100635■ Frontal cortex A2A SCH442416 Striatum 5-HT1B AZ10419369 Hypothalamus nAChR α4β2 ZW-104 Thalamus * 5-HT2A MDL-100907 Frontal cortex nAChR α7 Methyllycaconitine Hypothala
Slide 78: Receptor Occupancy using radio or non-radiolabelled tracer
Receptor Occupancy using radio or non-radiolabelled tracer R a d io la b e lle d r a c lo p r id e a s t r a c e r N o n - r a d i o la b e lle d r a c lo p r i d e a s tr a c e r 100 100 %D2 receptor occupancy %D2 receptor occupancy 80 80 60 60 40 40 20 20 0 0 0 .0 0 1 0 .0 1 0 .1 1 10 0 .0 0 1 0 .
Slide 79: Single and multi target receptor occupancy assay
Single and multi target receptor occupancy assay % 5-HT 1A receptor occupancy % 5-HT 2A receptor occupancy 100 ED50 = 10.92 mg/kg 100 ED50 = 0.03 mg/kg 100 ED50 = 0.11 mg/kg % D2 receptor occupancy 75 75 75 50 50 50 25 25 25 0 0 0 0.03 0.10 1.00 3.00 6.00 0.01 0.03 0.10 0.30 1.00 3.00 6.00 0.01 0.03
Slide 80: PK/PD: Prediction of Human exposure-RO Relationship
PK/PD: Prediction of Human exposure-RO Relationship 1. Predict Human PK Simple Emax model characteristics Parameter Value E0 (%) 5.6 EC50 (ng/mL) 4100 2. Assume PK in plasma is proportional to PK in Emax 75 brain gamma 1.3 3. Determine rat PKPD relationship 4. Assume human PKPD relationship is simil
Slide 81: In-vivo Brain Microdialysis Capabilities
In-vivo Brain Microdialysis Capabilities Study Designs Parallel Treatment Groups Cross-over Treatment Design Mechanism of Action Studies using various Antagonists/Blockers. Pharmacokinetic, Pharmacodynamic (PK/PD) Studies in-vivo, by simultaneous monitoring of neurotransmitters and drug concentratio
Slide 82: In-vivo Brain Microdialysis Facility
In-vivo Brain Microdialysis Facility Company Confidential 82 Copyright © 2016 Suven Life Sciences Limited
Slide 83: Neurotransmitter Measurement Using In-vivo Brain Microdialysis in rats
Neurotransmitter Measurement Using In-vivo Brain Microdialysis in rats Acetylcholine: SB-277011A, 10 & 30 mg/kg,s.c. - mPFC Glutamate: SB-271046 , 2.5 mg/kg,s.c. – Frontal cortex 400 350 Vehicle * ** SB-277011A 10 mg/ kg s.c. 300 ** SB-277011A 30 mg/kg, s.c. Acetylcholine (% Change) 250 * 200 * ** 1
Slide 84: Monitoring Neurotransmitters using In Vivo Brain Microdialysis
Monitoring Neurotransmitters using In Vivo Brain Microdialysis Serotonin (5-HT) Norepinephrine & Dopamine Fenfluramine 10 mg/ kg, i.p. (Guinea Pig- striatum) Atomoxetine 3 mg/ kg, i.p. ( Rat- prefrontal cortex) 600 800 Serotonin: Fenfluramine Dopamine: Vehicle Norepinephrine: Vehicle Dopamine: Atomo
Slide 85: Monitoring Endogenous Peptides using In Vivo Microdialysis
Monitoring Endogenous Peptides using In Vivo Microdialysis Substance P Prostaglandin E2 (PGE2) Formalin (5%), 50 µL ( Rat- Dorsal Horn) Formalin (5%), 50 µL ( Rat- Dorsal Horn) 300 600 Substance P: Vehicle PGE2: ELISA PGE2 (% change from mean basal value) Substance P: Formalin (5 %, 50 µL) PGE2: LC-
Slide 86: PK/PD Studies in Rats Using In-vivo Brain Microdialysis
PK/PD Studies in Rats Using In-vivo Brain Microdialysis / CSF Pharmacokinetics Dialysate profile and simultaneous Acetylcholine changes after GSK189254 400 24 Modularion in cortical Aetylcholine 350 Dialysate conc of GSK189254 20 300 16 Acetylcholine (% of baseline) GSK189254 conc (nmol) 250 12 200
Slide 87: In-Vivo Electroencephalography at Suven
In-Vivo Electroencephalography at Suven Infrastructure Telemetry Systems with Implantable Telemetry- Data Sciences International (DSI) Acquisition Platform- Ponemah Data Analysis Software- NeuroScore v3.0 with automated sleep scoring, seizure detection, video synchronization and batch processing
Slide 88: In-Vivo Electroencephalography Facility
In-Vivo Electroencephalography Facility Company Confidential 88 Copyright © 2016 Suven Life Sciences Limited
Slide 89: Modafinil: Dose Dependent Increase in Wakefulness
Modafinil: Dose Dependent Increase in Wakefulness Results Generated at Suven Edger and Seidel 1997 Results Generated at Suven are Comparable to the Results Reported in Literature *p<0.05, **p<0.01, ***p<0.001 Vs Vehicle Company Confidential 89 Copyright © 2016 Suven Life Sciences Limited
Slide 90: In-vivo Efficacy Pharmacology & Safety Pharmacology
Suven Discovery Research In-vivo Efficacy Pharmacology & Safety Pharmacology Company Confidential Copyright © 2016 Suven Life Sciences Limited
Slide 91: Therapeutic areas
Therapeutic areas • Cognition • Depression • Psychosis • Parkinson’s Disease • Anxiety • Pain • Obesity Company Confidential 91 Copyright © 2016 Suven Life Sciences Limited
Slide 92: Rodent Models of cognition
Rodent Models of cognition • Novel object recognition task (Scopolamine & Time induced episodic memory deficit) • Morris water maze (Scopolamine induced spatial memory deficit) • Radial arm maze (Scopolamine induced working memory deficit) • T-Maze (Scopolamine induced working memory deficit) • Fear
Slide 93: Rodent Models of Depression
Rodent Models of Depression • Dominant submissive assay • Forced swim test (Rat & Mice) • DRL-72s • Tail suspension test Company Confidential 93 Copyright © 2016 Suven Life Sciences Limited
Slide 94: Rodent Models of Psychosis
Rodent Models of Psychosis • MK-801 and Amphetamine induced hyperlocomotion & stereotypy • Prepulse inhibition • Condition avoidance response • Dominant submissive assay (Mania) • Resident Intruder Test (Aggression) Company Confidential 94 Copyright © 2016 Suven Life Sciences Limited
Slide 95: Rodent Models of Anxiety
Rodent Models of Anxiety • Elevated Plus Maze • Vogel conflict test • Hole board • Novelty induced hypophagia Company Confidential 95 Copyright © 2016 Suven Life Sciences Limited
Slide 96: Rodent Models for Pain Disorders
Rodent Models for Pain Disorders • Formalin Induced Nociception, Hot Plate & Acetic acid • Capsaicin induced mechanical allodynia induced writhing • Osteoarthritis & Rheumatoid arthritis induced pain • Complete Freund’s Adjuvant-Induced Mechanical models Hyperalgesia • Reserpine induced Myalgia in r
Slide 97: Rodent Models for Obesity
Rodent Models for Obesity • Acute food intake in adapted rats • Diet induced obesity (Rat & Mice) • Visceral sickness test • Behavioral satiety sequence Rodent Models for Parkinson's Disease • MPTP model in C57 mice Company Confidential 97 Copyright © 2016 Suven Life Sciences Limited
Slide 98: CNS Safety Pharmacology
CNS Safety Pharmacology (as per ICH S7 safety guidelines) Compound Used For Test Species Standardisation IRWIN test / Functional Rat or Mouse MK-801 and Diazepam Observation Battery (FOB) Rota Rod Rat or Mouse Chlordiazepoxide Open Field (Locomotor Activity) Rat or Mouse MK-801 and Haloperidol Proco
Slide 99: CV, GI, Renal & Respiratory Safety Pharmacology
CV, GI, Renal & Respiratory Safety Pharmacology (as per ICH S7 safety guidelines) Compound Used For Test Species Standardisation CV Safety QT, QTc, RR interval and Heart rate, Mean arterial blood Anesthesized Terfenadine pressure, systolic blood pressure Guinea pig and diastolic blood pressure GI Sa
Slide 100: Preformulation, Biopharmaceutics and Early Formulations
Suven Discovery Research Preformulation, Biopharmaceutics and Early Formulations Company Confidential Copyright © 2016 Suven Life Sciences Limited
Slide 101: Preformulation
Preformulation Screening • Salt and co-crystal screening • Crystallization screening • Polymorph screening Selection • Hygroscopicity • Solubility • Chemical stability • Physico Chemical Parameters (Log D, Log P, pKa etc.,) • Hydrates / Solvates Solid State • Thermal properties • Surface morphology,
Slide 102: Salt form screening and selection
Salt form screening and selection Continue crystallization attempts No Crystallinity Yes Unacceptable Yes Hygroscopicity No Solubility enhancement No Solubility Yes Stability enhancement No Stability Yes Salt for PK study Final salt candidate No Polymorphism Secondary candidate Yes No Control Salt f
Slide 103: Crystal Forms
Crystal Forms Crystalline Screening Compound from Chemistry Amorphous Screening Agglomeration Minimum Maximum Mean Crystal habit tendency Length(μm) 23.1 54.6 36.28 No Sticks Width (μm) 8 12.18 10.18 Company Confidential 103 Copyright © 2016 Suven Life Sciences Limited
Slide 104: Powder properties
Powder properties • Powder properties are measured using USP bulk density apparatus and angle of repose is calculated by funnel method • Bulk density Flow Property Angle of repose Carr’s Index Hausner Ratio Excellent 25-30 <10 1.0-1.11 • Tapped density Good 31-35 11.0-15.0 1.12-1.18 Fair 36-40 16-20
Slide 105: Compatibility studies
Compatibility studies Physical mixture of drug excipient ratio was taken which Represents to final formulation Analysis of mixture was carried out at regular intervals Physical Changes Chemical Changes (by DSC, IR etc.,) (by HPLC) Compatible / Incompatible Company Confidential 105 Copyright © 2016 S
Slide 106: Solubility & Stability Studies
Solubility & Stability Studies • Solubility • Solubility in different pH buffers • UV profile • Thermal properties (DSC & TGA) • Solution state stability • Hydrolysis (water, pH buffers 2, 4, 6 and 8) • Acid stress condition (0.01N HCl, 0.1N HCl) • Oxidation (peroxide oxidation at pH 2 and 8) • Soli
Slide 107: Formulations for Preclinical studies
Formulations for Preclinical studies • Vehicle selection and exclusion for PK and PD Studies • High dose formulations for early Tox studies • Solutions and Suspensions formulations for Rodent and Non rodents • Solubilization • Complexation • Enabled formulations • Dose formulation analysis • Formula
Slide 108: Vehicle selection & formulation preparation
Vehicle selection & formulation preparation Vehicle selection Physicochemical & Evaluation Formulation support Pharmacokinetic Pharmacology studies Toxicology studies studies (Activity / Efficacy) Biopharmaceutical Evaluation Developability Assessment Company Confidential 108 Copyright © 2016 Suven
Slide 109: Vehicle selection decision tree in preclinical studies
Vehicle selection decision tree in preclinical studies Stage 1 Aqueous Vehicles Water , PBS, Pharmasolve (i.v.) Stage 2 Suspending agents (p.o) HEC,CMC & Methyl cellulose Stage 3 Complexing agents (p.o & i.v) HPBCD ,Captisol Co-solvents (p.o & i.v) Stage 4 PEG 400 & Propylene glycol Stage 5 Surfacta
Slide 110: Biopharmaceutics
Biopharmaceutics • Vehicle selection and exclusion for in-vivo studies • Formulations for PK, Efficacy and Tox studies • Study of morphological changes in slurry formulations of Tox studies • Study of affect of stress like milling etc., on NCE during formulations • Biopharmaceutics characterization
Slide 111: Biopharmaceutics Risk Assessment
Biopharmaceutics Risk Assessment • Solubility in SGF & SIF • Solubility in FaSSIF & FeSSIF fluids • pH Solubility Profile • Stability in GI fluids • Chemical stability • Effect of food on bioavailability • Solubility estimation • Permeability evaluation • Absorption issues in PK studies Solubility a
Slide 112: Drug Absorption Studies
Drug Absorption Studies • In situ single/double/triple pass method • In situ closed loop method • Everted gut sac method • Intestinal rings method • Intestinal diffusion method • Segmental dependent perfusion studies • pH dependent perfusion studies • Mesentric blood sampling method • Pre hepatic &
Slide 113: Biopharmaceutics Evaluation
Biopharmaceutics Evaluation Disintegration Dissolution Absorption Elimination Dissolution rate Permeability Solubility limited limited limited Drug molecules Body Tablet Suspension Solution I.V. injection Dissolution rate X Solid X Solid X Solid Solubility MRT disint MRT diss MRT abs MRT iv Solid So
Slide 114: Early Formulations
Early Formulations “API – Focused” Characterize and Monitor API Properties to Enable Strong Drug Product Development • Physical • Chemical • Mechanical • Biopharmaceutical Minimal Drug-Product Development • “Fit-for-use” formulation that meets the needs for early clinical evaluation • “Fit-for-use”
Slide 115: Early Formulations
Early Formulations Exploratory Formulation Approach Use the simplest possible formulation consistent with the clinical goal of the early phase studies • API only: “Powder in a bottle or capsule approach” • “Exploratory formulations” The goal is to achieve adequate exposure from a “dosage form” of ad
Slide 116: Oral formulations with varying degrees of complexity
Oral formulations with varying degrees of complexity Increasing Complexity API in Bottle - solution preparation in clinic API API in Capsule only API in Bottle - suspension prep in clinic Solution Aqueous suspension Simple Formulated Capsule Exploratory Simple Formulated Tablet Formulations Solubili
Slide 117: Phase 1 & Phase 2A Formulations
Phase 1 & Phase 2A Formulations • Formulation for Phase-I & II studies • Dosage form Selection • Immediate Release Formulations • Solution, Suspensions • API-in-Capsules and Tablets • Taste masking formulations • Solid Dispersions • Clinical supplies manufacturing • eCTD IND documentation Company Co
Slide 118: Novel Formulations
Novel Formulations • SEDDS / SMEDDS Formulations • Nano Suspension Formulations • Microemulsion Formulation • Micro Capsules • Nano Crystal formulation • Liposomes • Extended Release Formulations • Timed Release Formulations Company Confidential 118 Copyright © 2016 Suven Life Sciences Limited
Slide 119: Formulation Analysis Support
Formulation Analysis Support • Analytical support to drug product development, clinical supplies manufacturing and stability studies • State of art HPLC instrumentation lab with six Agilent HPLCs having various detectors of UV, PDA, RI and ELSD • Dissolution testing systems equipped with 12+2 and 6+
Slide 120: Discovery Toxicology Services
Suven Discovery Research Discovery Toxicology Services Aggressive Timelines together with high Scientific Quality Decision making for NCE progression Company Confidential Copyright © 2016 Suven Life Sciences Limited
Slide 121: Suven Life Sciences introduces 4-Day Tox in 14 Days, 7-Day Tox in 17 Days
Suven Life Sciences introduces Wherein Toxicity studies are conducted on new chemical entities in the early stage of discovery. Data are generated in extremely rapid timelines at the same time with highest quality and scientific integrity for your decision making of molecule progression. 4-Day Tox i
Slide 122: Discovery Toxicology Capabilities
Discovery Toxicology Capabilities Toxicity studies Acute toxicity studies in rodents 4-Day / 7-Day repeat dose discovery tox studies in rodents Repeat dose (14 / 28 days) toxicity studies in rodents In vivo & In vitro Genotoxicity studies Bacterial reverse mutation test (Ames Test) In vitro
Slide 123: Discovery Toxicology - Facilities
Discovery Toxicology - Facilities Animal Facility Vivarium spreading over 6000 sft Double corridor system HEPA filtered 100% clean air supply Individually ventilated cages Necropsy room State of art instruments Clinical Pathology Lab Histopathology Lab Documentation Room Well Equipped Genotox Lab Sp
Slide 124: Discovery Toxicology – Good Research Practices
Discovery Toxicology – Good Research Practices SOPs Planning and initiation of study, live phase of toxicity study, necropsy and organ collection, clinical and histopathology, training of personnel, archiving of study data and specimens, equipments Formats Logbooks Individual training record
Slide 125: Advantages with Suven Discovery Toxicology
Advantages with Suven Discovery Toxicology Aggressive timelines together with high scientific excellence and quality data are critical in drug discovery. Having years of experience in discovery research, Suven Tox offers quality results of preclinical toxicity studies. The tox division is well equip
Slide 126: Discovery Toxicology
Discovery Toxicology Suven Discovery Tox expertise and contribution to progression of NCEs Identified phospholipidosis in early stage of discovery by conducting 4-day tox Identified potential platform toxicity Identified potential back-up compounds with good margin of safety Contributed towards crit
Slide 127: Contacts
Contacts: Venkat Jasti Chairman & CEO E-mail: [email protected] Ramakrishna Nirogi Vice President, Discovery Research E-mail: [email protected] Company Confidential 127 Copyright © 2016 Suven Life Sciences Limited