David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 INTRODUCTION SIGNAL TRANSDUCTION KINASES ARE IMPORTANT IN HUMAN BIOLOGY/DISEASE: The process describing how a signal (chemical/physical) is transmitted through a - Kinases are a superfamily of proteins (5th largest in humans) cell ultimately resulting in a cellular response - 518 genes and 106 pseudogenes - Diverse in size, subunit structure, and cellular location RECEPTOR TRANSDUCERS EFFECTORS - ~260 residues make up their conserved catalytic core - dysregulation of kinases occurs in many diseases (cancer, inflamatory, degenerative, and autoimmune diseases) - signals can originate inside or outside the cell - 244 of the 518 map to disease loci - signals generally passed through a series of steps (signal transduction pathway) often consisting of multiple enzymes and messengers protein O - pathways open possibility for signal aplification (>1x106) kinase ATP + PROTEIN OH ADP + PROTEIN O P O - Extracellular signals transduced by RTKs, GPCR’s, guanlyl cyclases, or ligand-gated Ion channels O - Phosphorylation is the most prominent covalent modification/signal in KINASES INHIBITORS ARE IMPORTANT IN DISEASE THERAPY/RESEARCH: cellular regulation - currently 51 FDA approved small molecule kinase inhibitors - “converter enzymes” (protein kinases and phosphoprotein phosphorlyases) - 4 FDA approved antibody kinase inhibitors are regulated; conserve ATP/maintain desired target protein state - thousands of known inhibitors spanning the kinome - pathway ends by affecting biomolecule function or gene expression - used to treat a broad range of disease types; primarily cancer - inhibitors used as tools for biological discovery

A Brief History - 1984: Staurosporine first kinase inhibitor discovered (non-selective) - 1991: first 3D structure of kinase solved (PKA) - 1999: First FDA approved antibody kinase inhibitor (Trastuzumab, Herceptin™; Genentech) - 2001: First FDA approved small molecule kinase inhibitor (Imatinib, Gleevec™; Novartis) Protein Kinases: GROUP MEETING INCLUDES: H - generally highly specific often recognizing consensus sequences in their - Introduction to signal transduction O N target (contains target residue) - Introduction to therapeutically relevant kinase targets - can phosphorylate one or many residues on a protein - Introduction to kinase structure - residues phosphorylated bear a free hydroxyl/phenol (serine, threonine, - Case studies in targets and their inhibitors tyrosine) - Introduction to drug resistance to kinase inhibitors - tyrosine kinases only exist in multi-cellular organisms − important for - Introduction to drug discovery in kinase inhibitors N N intercellular communication - Introduction to perspectives on the future of kinase O - usually locked in their “inactive conformations” − blocked by intrasteric Me control/pseudosubstrates inhibitors as therapeutics 2+ - Summary of all FDA approved kinase inhibitors OMe - turned on by interaction with messengers (cyclic nucleotides, Ca , etc.) or phosphorylation/dephosphorylation HN GROUP MEETING DOES NOT INCLUDE: Me - tyrosine kinases broken into two broad categories (RTK’s and NRTK’s) - an exhaustive discussion of any of the above topics Staurosporine - 90 tyrosine kinases - 58 RTK’s and 32 NRTK’s David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 KINASE-MEDIATED SIGNAL TRANSDUCTION PATHWAY: THERAPEUTIC TARGETS 1. Receptor binding - results in conformational change/dimerization of RTK 2. Phosphorlyation on RTK - (trans)autophosphorylation via now active kinase Below is a list of clinically validated kinase targets: 3. Recognition of phosphoprotein - SH-2 domain binds phospho-RTK BCR-Abl - fusion kinase in some cancers; NRTK; activates numerous pathways 4. Transphosphorylation - newly bound protein is phosphorylated cKit - RTK; activates numerous MAP-Kinase pathways 5. Cascade continues - successive recognitions and phosphorylations EGFR - epidermal growth factor receptor; also ErB1; RTK 6. Effector reached - cascade ends by modulating final target and its function VEGFR - vascular endothelial growth factor receptor; RTK PDGFR - platelet derived growth factor receptor; RTK ErbB - (also called HER) other RTK’s important in regulating growth ALK - anaplastic lymphoma kinase; NRTK B-raf - Ser/Thr kinase; MAP/ERK pathway JAK - Janus kinase; NRTK; part of JAK-STAT pathway BTK - Bruton’s tyrosine kinase; important in B-cell development MET - cMET or HGFR; RTK; activates numerous pathways CDK - cyclin-dependent kinase; Ser/Thr kinase; help regulate cell cycle Rho - also ROCK; Ser/Thr kinase; associated with degenerative diseases TRK - tropomyosin receptor kinase; RTK; activates MAP and PI3 pathways PROTEIN KINASE CLASSES: Syk - spleen tyrosine kinase; NRTK 1. Ser/Thr Protein Kinases a. Cyclic nucleotide dependent Flt3 - also CD135; RTK; one of most frequently mutated genes in AML b. Ca2+-calmodulin dependent (CaM) PI3K - phosphoinositol 3 kinase; lipid kinase; PI3 involved in diverse signaling c. Protein kinase C (PKC) d. mitogen activated protein kinases (MAP) FKBP/mTor - mammalian target of rapamycin; Ser/Thr kinase; FKBP e. G-protein-coupled/Rhodopsin/BARK association necessary for activity 2. Ser/Thr/Tyr Protein Kinases a. MAP kinase kinase (MAPK) 3. Tyr Protein Kinases - TARGETS MUST BE LINKED TO PHENOTYPE a. Cytosolic tyrosine kinases - most targets are oncogenes discovered by clinical/biochemical investigation b. Receptor tyrosine kinases (RTKs) - some above targets are markers for certain cancer (or disease) types - not all kinase targets are oncogene products Kinome - the complete set of kinases encoded in a genome - all kinase targets are involved in cell cycle/cell growth regulation Protein Kinase - enzyme that transfers a phosphate to a target protein in an ATP- - many are in or are upstream of MAP-kinase pathways (stimulate cell division) dependent manner - certain disease states demonstrate activated kinase forms (abnormally Phosphoprotein Phosphatase - enzyme that catalyzes the hydrolysis of phosphates active or constitutively) from phosphoproteins Intrasteric control - regulation of enzyme activity in which an internal sequence of - some cancer cells rely heavily on the above targets/pathways protein resembling substrate (pseudosubstrate) binds the active site, - makes them particularly sensitive to inhibition - therapeutic index blocking activity - some inhibitors target a single kinase (selective/mono-specific) Receptor Tyrosine Kinase (RTK) - membrane-bound tyrosine kinase who’s activity is - some inhibitors target multiple kinases (poly-specific) dictated by interaction with an extracellular stimulus Non-receptor Tyrosine Kinase (NRTK) - completely cytosolic tyrosine kinase Mitogen - a chemical substance that encourages cell division (mitosis) Oncogene - a gene that has the potential to cause cancer via mutation/overexpression SH-2 domain - conserved domain among intracellular signal transducing proteins; Proto-oncogene - a gene in its normal state that could become an oncogene recognizes phosphotyrosine Phenotype - physiological/observable characteristics of an organism David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 KINASE STRUCTURE [overlay of active/inactive kinase] [depiction of C- and R- spines]

- activation usually result of phosphorylation - the “spines” are linear arrangements or binding a “signal” of hydrophobic residues - activation changes DFG orientation (below) - formation necessary for function - functions validated by mutation - ACTIVE = AS open; !C in; DFG-Asp in - some tolerance R-spine

gatekeeper residue IMPORTANT STRUCTURAL MOTIFS: N-Lobe - N-terminal small lobe of mostly β-sheet ACTIVE INACTIVE C-Lobe - C-terminal large lobe of mostly !-helix Hinge - short portion connecting the N- and C-lobes; important for inhibitor binding INHIBITOR BINDING MOTIFS: K/E/D/D - signature motif defining functional kinases (stabilizing/active-site residues) - figure at left depicts pockets/regions Activation Segment - regulatory loop that changes conformation upon activation generally utilized by inhibitors - most inhibitors make 1-3 hydrogen DFG - three residue motif at beginning of activation segment; large role in catalysis bonds w/ hinge residues C-Spine - 8 hydrophobic residues that facilitate ATP binding - specificity defined largely by alternative R-spine - 4 hydrophobic residues that position substrate appropriately pockets Shell residues - 3 residues surrounding/stablizing the R-spine - gatekeeper residue size determines access to hydrophobic pocket II SH2 (Gatekeeper) - shell residue controlling access to an important binding pocket - binding active vs. inactive state !C-helix - dynamic motif whose conformation dictates binding of some inhibitors provides access to different pockets David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 INHIBITOR TYPES BCR-Abl - BCR-Abl is a fusion gene (hybrid of two previously separate genes) - two original genes normally exist on two different chromosomes - expression of fusion results in a constitutively active Abl kinase - first discovery of a genetic Philadelphia Chromosome abnormality directly associated with a cancer - decreases apoptotic potential and activates MAP kinase pathways at Ras - associated with various leukemias (CML and ALL) TYPE I - Binds in ATP binding pocket; active form; DFG-Asp IN; - first clinically validated kinase Type I 1/2 - Binds in ATP binding pocket; inactive form; DFG-Asp IN target; target of imantinib TYPE II - Binds in ATP binding pocket; inactive form; DFG-Asp Out TYPE III - Allosteric inhibitor binding near ATP site HINGE TYPE IV - Allosteric inhibitor binding away from ATP site adenine solvent pocket TYPE V - bivalent inhibitor spanning two regions region TYPE VI - covalent inhibitors; partially fall into types I and II O N

INHIBITOR BINDING, SPECEFICITY, AND IMATINIB N H MeO CN N N HN OMe N H N H N N Cl Cl Bosutinib (Bosulif, Wyeth) 2012, CML N hydrophobic increased cellular pocket activity O R 1. N-Me-piperazine, NaH O NO2 O NH N H 2. H , Pd/C 2 N N NH 2 Cl N MeO MeO N Cl Cl provided Tyr- O kinase activity hydrophobic allosteric Vaidyanathan, OPRD 2013, N OMe 1, HC(OEt)3 pocket specific 500. H N NMe N CN HINGE Me pocket O R 1 H H O N O N HN N N NH N N N POCl N O N MeO CN 3 N HN N-Me-piperazine Me MeO NC HN OMe MeO NH O improved solubility/ abolished PKC N oral bio-availability adenine Imatinib (Gleevec, Novartis) activity H pocket 2001; Brc-ABL N Cl Cl Cl Cl David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 GROWTH FACTOR RECEPTORS MEK1/2 (EGFR, VEGFR, PDGFR) F I ACHN: 9.8 nM - MEK is a midstream - All are closely related receptor tyrosine kinases HT-29: 0.57 nM - All have ability to activate multiple pathways (depends upon phosphorylation part of Ras-MAP pattern) - MAPK, PI3K, STAT O HN pathway - oncogenes generally - each has multiple subtypes and vary by receptor and tissue distribution Me - variably associated with certain types of cancers N N upstream - when implicated, overexpression is usually the cause of aberrant activity - activation of ERK - EFGR one of the most commonly drugged kinases O N O signals for cell proliferation - the target of the antibody kinase inhibitors Me - 4-aminoquinazoline is everywhere!! - MAP pathways implicated in many - in 6 FDA-approved EGFR inhibitors Ac HINGE N cancer types H O N adenine MeO Trametinib (Mekinist, GSK) Kawasaki, ACS Med. pocket Chem. Lett. 2011, 320. MeO N hydrophobic 2013; melanoma O pocket solvent O O HN region O CH (CO H) , 1. POCl 2 2 2 N 3 N Erlotinib (Tarceva, Genentech) Ph Ac2O 2. MeNH2 2004; NSCLC, pancreatic cancer N N adenine H H O N O O N NHMe Me pocket Ph Ph HINGE N Br N Me ACHN: renal ACHN: 1270 nM diethyl adinocarcinoma HT-29: 100 nM malonate HN N N N HT-29: colorectal adenocarcinoma solvent O HN O OH region Me allosteric 1. TsCl, Et N O specific N 3 K2CO3 2. 4-Br-aniline N S pocket H N O N N O 2 O Pazopanib (Votrient, GSK) 96% O N N O Me 2009; RCC, soft tissue sarcomas Me Ph Me H2N NO2 t N NO2 N NO2 BuONO, Br Br AcOH HN MeO3BF4 MeN O HN Me O HN O HN Me Me Cl N Cl Me N N Me Me Me Cl N N SnCl2, N N N N HCl O N O H2N SO2NH2 2 3 N O N O O N O Me HCl, N N 1. NaHCO3, 2 N NH2 Me 2. MeI, Cs CO 3 MeN 2 3 MeN Pazopanib Br Li, Modern Drug Synthesis, ACHN: >3000 nM ACHN: >3000 nM ACHN: 830 nM Me Wiley, 2010. Me HT-29: >1000 nM HT-29: >1000 nM HT-29: 135 nM David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 OH 2- PI3K (phosphoinositide 3-kinase) HO 3 OPO3 BTK (Bruton’s Tyrosine Kinase) 4 5 O 1 2- O OPO3 - Extremely important kinase in B-cell Me O O P O OH development Me O O - targeted for lymphomas (B-cells are a type of O phosphatidylinositol (4,5) diphosphate - PI(4,5)P lymphocyte) 2 - low expression in many types of tissues - - PI3 kinases are a class of lipid kinases good therapeutic index - Type I: PI3Kẟ and PI3K" are therapeutically relevant - interest is high for treatment of autoimmune - catalyze: PI(4,5)P2 ➜ PI(3,4,5)P3 disorders (many ongoing clinical trials) - PIP3 is a signaling molecule that has multiple - many of the inhibitors of interest are covalent implications in cancer phenotypes - control numerous diverse cellular functions (growth, motility, differentiation, and intracellular trafficking) - many ongoing clinical trials targeting these enzymes - PI3K can be activated by numerous receptors N O N adenine N N pocket O HN N N HINGE Cys481 allosteric H2N N specific HN N adenine pocket pocket N Me hydrophobic pocket HINGE N Ibrutinib (Imbruvica, Pharmacyclics/J&J) hydrophobic 2013, Mantle cell lymphoma, CLL, macroglobulinemia pocket F O OPh OPh OPh H2NHN •2 HCl Idelalisib (Zydelig; Gilead) NHBoc NC CN N 2014; CLL, SLL, follicular lymphoma O DIPEA TMSCHN2 Et3N Cbz

NO2 1. (COCl) , PhNH NH2 2 2 HOBt, CDI NH Me OPh 2. Zn, HCO2NH4 H 4 NC NC OH N H O Cl OH OMe Ph N Ph CN CN F O F O NH2 Ar N N F O NH2 Ar N 1. P(OH)2/C, H2 NC HN N 1. TFA N 2. NHBoc COCl N AcOH N O HN 2. ZnCl2, N N N DIPEA, N O ZnCl, N N NH H2N HMDS 6-Cl-purine Idelalisib NH Me OH Me H NH2 H N N CbzHN 4 Ph CbzN Hughes, OPRD 2016, 1855. F O Moturu, RSC Avd. 2018, 15863. O David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 JAK (Janus kinase) CDK4/6 - cyclins are a number of small proteins - The beginning of the JAK-STAT pathway involved in regulating cell cycle - STAT = signal transduction activator of - different cyclins pair w/ different CDK’s transcription to activate different cell stages - 4 types (JAK1, JAK2, JAK3, TYK2) - activated CDK’s trigger changes in - JAKs are receptor-associated kinases transcription to enter next phase (often cytokine receptors; immune system) - CDK4/6 phosphorylate a repressor - single point mutations are often associated allowing S phase to start with proliferative effects/autoimmunity - “R” = checkpoint where cells can trigger - associated with leukemias, psoriasis, apoptosis (programmed cell death) rheumatic diseases - other CDKs are of interest as therapeutic - kinase target of the first non-cancer targets approved kinase inhibitor hydrophobic - continues to be an active area of research pocket for a variety of disease types adenine HN pocket Me Me N N O Tofacitinb (Xeljanz, Pfizer) N allosteric 2012; rheumatoid arthritis specific solvent JAK3 pocket region N N N N O O allosteric H specific ribose N pocket pocket hydrophobic HINGE Me pocket N Palbociclib (Ibrance, Park Davis) adenine 2015; ER2+ and HER2+ breast cancer pocket Me N Me Pd(OAc)2, Et N, N Br 3 Br Et3N, 5; N N cyclopentylamine H N Ac2O N Cl N Cl Cl N NH Cl N N O HINGE Me Me Me O BnCl O NaBH4 O Me BocN NH CO H 2 N NBn NBn 2 NBS MeO N MeO N MeO N 5 6 Me H H Cl H Br Cl Rh(cod) OTf, t 2 97% cis N NC 1. Pd(OAc)2, DIPEA, Me P( Bu)2 ligand, 66% ee Cy2P N DPEPhos, 7 CyMgBr, Fe H2 (200 psi) HN N N O Br Me 2. HCl 6 N O N Palbociclib N Me OH Ts N Cl N N O ligand O Me - resolution w/ (L)-DTTA Tofacitinb Me NBn to upgrade amine ee N 7 Carvalho, Eur. J. Org. Chem. 2019, 615. H R Duan, OPRD 2016, 1191. David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 DRUG RESISTANCE COVALENT INHIBITORS O O - treatment of cancer with kinase inhibitors applies a selection pressure to the O O - heightened interest in covalent cells it affects X S inhibitors R R R F - cancer cells divide unchecked and are prone to further mutation - shown they can be selective and safe Cys/Tyr; Irrev. - resistance to kinase inhibitors generally arises rapidly (a few months to a year) Cys; Irrev. Cys; Irrev. - limited to kinases w/ accessible O - type of resistance varies between type of cancer treated and drug used O O residues - biggest drawback to this type of therapy S R Ph - benefits include: prolonged N N pharmacodynamics, high potency, R H TYPES OF RESISTANCE: Cys, Lys, His; Irrev. O rapid validation A. Target Dependent Resistance Met; Irrev. F - drawback of straightforward resistance 1. target protein point mutation O O - much discussion about “kinetic 2. target protein amplification/overexpression selectivity” B. Target Independent Resistance Het O R CN - some solutions: reversible covalent 1. decreased drug uptake Lys; Irrev. O & metabolically limited warheads 2. increased drug efflux O - opportunities may lie in targeting 3. upregulation of alternative/ R other residues (non-Cys) OMe downstream pathways R CN - list at left are a few examples of explored warheads and targeted Cys; Irrev. Cys; Rev. CASE STUDY: IMATINIB O residues - target point mutation is the prominent resistance mechanism (limited) - >100 different point mutations have been described in resistant patients GOING FORWARD - only ponatinib is active against imantinib resistant cancers (gatekeeper) - still active areas of research - academia and industry - four mutations account for 60% of the resistant mutants - new cancer targets include: TAM kinases and other CDK’s 1. Thr-315-Ile (gatekeeper mutation) - potential synergies with immunotherapy 2. Tyr-253-Phe (changes in van der Waals forces) - PROTACs w/ kinase inhibitors being investigated 3. Glu-255-Lys/Val (inside ATP binding pocket) - high interest in entering disease areas outside of cancer 4. Met-351-Thr (!E-helix of C-lobe; mechanism unclear) - selectivity becomes paramount in treatment of non-terminal diseases - necessity of/role of selectivity debated in kinase inhibitor cancer treatment DRUG DISCOVERY OTHER DISEASE AREAS Solved Problems: 1. Autoimmune disease - 20 JAK inhibitors in clinical trials for autoimmune disorders - poster child of rational drug design Thermal Proteome Profiling - thousands of kinase structures known - generating biological models is largest challenge - much known about kinase binding 2. Degenerative disease - kinase inhibitors historically found via - challenging; poor of understanding disease mechanism HTS or fragment-based approaches - chronic unfolded protein response mediated by kinases 3. Infectious disease Current/Future Challenges: - Plasmodium falciparum-selective kinases - finding novel hinge binders 5. Angiogenesis - rapidly evaluating whole-cell kinase - systemic VEGFR inhibitors for macular selectivity degeneration (intravitreal injection only other option) - creating proper biological models point mutation - change of a single nucleotide in the gene sequence that leads to a New Technologies: different amino acid in the expressed protien gene amplification - an increase in the number of copies of a gene relative to the rest of - Activity-based protein profiling (ABPP) rapidly generates kinome coverage data the genome - Thermal proteome profiling (TPP) generates data about other off-targets overexpression - a change in transcriptional regulation resulting in more protein being - computational techniques (FEP and WaterMap) helps predict & rationalize SAR expressed David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19

Me O N O N Me N H N N N N MeO CN N N N O H H N HN OMe N N N N H N NMe2 Imatinib (Gleevec, Novartis) Bosutinib (Bosulif, Wyeth) Cl Cl HN Ribociclib (Kisqali, Novartis) 2001; CML, ALL, CEL/HES, GIST, DFSP, mastocytosis 2012, CML 2017, ER2+ and HER2− breast cancer BCR-Abl, Kit, PDGFR BCR-Abl, Src, Hck, Lyn CDK 4/6

O Cl N S NH F H Me O OH N N O Cl O H S N N N N O N N N HN N O F N Me N N Me H HN Me Me Dasatinib (Sprycel, BMS) Vemurafenib (Zelboraf, Genentech) Palbociclib (Ibrance, Park Davis) 2006; CML 2011; melanoma (V600E) 2015; ER2+ and HER2+ breast cancer BCR-Abl, (PD)(E)GFR, Src, Kit, EphA2, Lck A/B/C-raf CDK 4/6 tBu F N F O H Me N S N N S F Me H N Me Me O N N N N N N N H F N N N O Me N NH2 H N Me N Nilotinib (Tasigna, Novartis) Abemaciclib (Verzenio, Eli Lilly) CF3 Dabrafenib (Tafinlar; GSK) 2017; HER2+/− breast cancer F 2007; CML 2013; melanoma BCR-Abl, PDGFR, DDR1 B-raf CDK4/6 Me Me N NH Me O Me N 2 N O H N H N N N N O N MeO N N Me F3C N H F H N N O O Me Me Ponatinib (Iclusig, Ariad) NH Netarsudil (Rhopressa, Aerie Pharm.) Encorafenib (Braftovi, Array) 2012; CML, ALL Cl 2018; Glaucoma 2018; melanoma O S O BCR-Abl, (V)(PD)(E)GFR, Src Rho kinase B-raf Me David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19

Cl NMe2 Br O O Me N N N F N Me CN Me HN O Cl F F N N O HO HN N Me NC Me H N N NH2 N N O N N Crizotinib (Xalkori, Pfizer) N N Me H 2011; NSCLC O Me ALK, ROS1 Lorlatinib (Lorbrena, Pfizer) Tofacitinb (Xeljanz, Pfizer) 2018; NSCLC Binimetinib (Mektovi, Array) 2012; rheumatoid arthritis Me ALK 2018; melanoma JAK3 MEK1/2 Cl N O Me Me I EtO2S N N N N CN N CN H H HN O O S O Me HN N N H N N N N Me Me F Me Me O O N O Ceritinib (Zykadia, Novartis) 2014; NSCLC (crizotinib res.) N ALK, IGF-1R, ROS1 N O Trametinib (Mekinist, GSK) 2013; melanoma N N N N N MEK1/2 H H Me Me H F Ruxolitinb (Jakafi, Incyte) Baricitinib (Olumiant, Eli Lilly) N 2011; myelofibrosis, polycythemia vera N HO O 2018; rheumatoid arthritis JAK1/2 JAK1/2 H N HN I CN NH O Me O F N Acalabrutinib Alectinib (Alecensa, Hoffman-LaRoche) NH N (Calquence, Acerta Pharma) 2015; NSCLC F 2017; Mantle cell lymphoma ALK, RET N Cobimetinib (Cotellic, Genentech) N BTK O NH OMe 2015; melanoma 2 H MEK1/2 Me N N N OMe O N Me N N Cl MeO F O O N N N Me HN N O MeO N N N N N O N NH2 N Me P H H N Fostamatinib (Tavalisse, Rigel) Ibrutinib (Imbruvica, Pharmacyclics/J&J) Brigatinib (Alunbrig, Ariad) Me Me OPO3H 2017; NSCLC 2018; thrombocytopenia 2013, Mantle cell lymphoma, CLL, macroglobulinemia ALK, ROS1, IGF-R1, EGFR Syk BTK David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 MeO N N Me2N Me N NH O N F N OMe N O N O HN Cl SO2Me HN HN N N Gefitinib (Iressa, Astra Zeneca) NMe Lapatinib (Tykerb, GSK) H 2003/2015; NSCLC F + O O EGFR 2007; HER2 breast cancer EGFR; ErbB2 Cl Osimertinib (Astra Zeneca) 2015; NSCLC Me EGFR O N O N O MeO O N N MeO N O N O O HN Cl HN Me N N 2 HN Dacomitinib (Visimpro, Pfizer) Erlotinib (Tarceva, Genentech) 2018, NSCLC HN Cl F 2004; NSCLC, pancreatic cancer O EGFR EGFR Afatinib (Gilotrif, B-I) 2013; NSCLC F EGFR; ErbB2/4 Et O N H H O N N O CF3 O CN O O Cl O O Cl MeHN O NMe2 HN N N Lenvatinib (Lenvima, Eisai) N N H2N 2015; thyroid cancer H H Neratinib (Nerlynx, Puma) O (VE)(PD)(V)GFR, Kit, RET F 2017; HER2+ breast cancer MeO N Regorafenib (Stivarga, Bayer) ErbB2 Cl 2012; CRC (VE)(PD)(F)GFR, BCR-Abl, B-raf, Kit,RET

Me N H H N N N O O NH H O O Me N N O F N O MeO O MeO N HN N Cabozantinib (Cometriq; Exelixis) OMe Vandetanib (Caprelsa, Astra Zeneca) 2012; Thyroid cancer, RCC, HCC Me 2011; Thyroid cancer F Br MeO N RET, MET, Kit, VEGFR, Flt3, Azl, TrkB (V)EGFR, RET, Src, Brk, EphR Nintedanib (Ofev/Vargatef, B-I) 2014; idiopathic pulmonary fibrosis VEGFR, PDGFR, FGFR David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19 H O CF3 Me N O H N O Cl H2N O MeHN O S N N N N MeHN S O N Me N N N N N Me H H Pazopanib (Votrient, GSK) Me Axitinib (inlyta, Pfizer) Sorafenib (Nexavar, Bayer) 2005; thyroid cancer, HCC, RCC 2009; RCC, soft tissue sarcomas 2012; RCC VEGFR, PDGFR, FGFR, Kit, Lck, Fms VEGFR, PDGFR VEGFR, PDGFR, B/C-raf, Kit, Flt3, RET

O F MeO Me N F O HN N HO H2N HN N N N Me F N N Me N N N H NH N N O Gilteritinib (Xospata, Astellas) N O 2018; AML Me Me N O Me H Larotrectinib (Vitrakvi, Bayer) Flt3 Sunitinb (Sutentm, Pfizer) 2018; solid tumors w/ NTRK fusions 2006; RCC, GIST, pancreatic NET TRK N PDGFR, VEGFR, Kit, RET, Axl, Flt3 HN N N N HN Idelalisib (Zydelig; Gilead) N O HN N N O 2014; CLL, SLL, follicular lymphoma PI3K OMe HN N Me N O N N N N Me OR H N OMe H N N NH2 Cl O Me Copanlisib (Aliqopa; Bayer) Me 2017; follicular lymphoma F O Duvelisib (Copiktra; Verastem) PI3K H 2018; CLL, SLL, follicular lymphoma O N PI3K O O OH N Me ALL = acute lymphoid leukemia O O O AML = acute myeloid leukemia O MeO HO DFSP = dermatofibrosarcoma protuberans Me CEL = chronic eosinophilic leukemia O OMe Me CLL = chronic lymphomic cancer N N Midostaurin (Rydapt, Novartis) CML = chronic myeloid leukemia O 2017; AML, mastocytosis GIST = gastrointestinal stromal tumor Me VEGFR2, PDGFR, Flt3, PKC HCC = hepatocellular carcinoma Me Me HES = hypereosinophilic syndrome OMe Binders of FKBP12/mTor NSCLC = non-small cell lung cancer N R = OH Rapamycin/Sirolimus (Rapamune, Wyeth) 1999 RCC = renal cell carcinoma Me R = dimethylolpropionic acid Temsirolimus (Torisel, Wyeth) 2007 SLL = small lymphocytic lymphoma O R = CH2CH2OH Everolimus (Afinitor, Novartis) 2009 David Peters Baran Group Meeting Kinase Inhibitors: An Introduction 2/2/19

Major References/Resources: Roskoski, Pharm. Research 2015, 100, 1. Roskoski, Pharm. Research 2016, 103, 26. Gray, Nat. Rev. Drug Discov. 2018, 17, 353. Shokat, Annu. Rev. Biochem. 2011, 80, 796. Wu, Trends in Pharm. Sci. 2015, 36, 422. Capdeville, Nat. Rev. Drug Discov. 2002, 1, 493. Flanagan, J. Med. Chem. 2014, 57, 5023. Sánchez-Martínez, Bioorg. Med. Chem. Lett. 2015, 25, 3420. Roskoski, http://www.brimr.org/PKI/PKIs.htm, 1/20/19. Garrett; Grisham, Biochemistry, Boston: Brooks/Cole, 2010. Ward; Goldberg, Kinase Drug Discovery, Cambridge: Royal Society of Chemistry, 2019.