PKIS deep dive yields a chemical starting point for dark kinases and a cell active BRSK2 inhibitor

Abstract The Published Kinase Inhibitor Set (PKIS) is a publicly-available chemogenomic library distributed to more than 300 laboratories by GlaxoSmithKline (GSK) between 2011 and 2015 and by SGC-UNC from 2015 to 2017. Screening this library of well-annotated, published kinase inhibitors has yielded a plethora of data in diverse therapeutic and scientific areas, funded applications, publications, and provided impactful pre-clinical results. GW296115 is a compound that was included in PKIS based on its promising selectivity following profiling against 260 human kinases. Herein we present more comprehensive profiling data for 403 wild type human kinases and follow-up enzymatic screening results for GW296115. This more thorough investigation of GW296115 has confirmed it as a potent inhibitor of kinases including BRSK1 and BRSK2 that were identified in the original panel of 260 kinases as well as surfaced other kinases that it potently inhibits. Based on these new kinome-wide screening results, we report that GW296115 is an inhibitor of several members of the Illuminating the Druggable Genome (IDG) list of understudied dark kinases. Specifically, our results establish GW296115 as a potent lead chemical tool that inhibits six IDG kinases with IC50 values less than 100 nM. Focused studies establish that GW296115 is cell active, and directly engages BRSK2. Further evaluation showed that GW296115 downregulates BRSK2-driven phosphorylation and downstream signaling. Therefore, we present GW296115 as a cell-active chemical tool that can be used to interrogate the poorly characterized function(s) of BRSK2.

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PKIS Deep Dive Yields a Chemical Starting Point for Dark Kinases and a Cell Active BRSK2 Inhibitor

10.1101/2020.06.15.153072 ◽

2020 ◽

Author(s):

Tigist Y. Tamir ◽

David H. Drewry ◽

Carrow Wells ◽

M. Ben Major ◽

Alison D. Axtman

Keyword(s):

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Clinical Results ◽

Deep Dive ◽

Downstream Signaling ◽

Starting Point ◽

A Cell ◽

Summary Statement ◽

Druggable Genome

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Screening of Tau Protein Kinase Inhibitors in a Tauopathy-relevant cell-based model of Tau Hyperphosphorylation and Oligomerization

10.1101/821389 ◽

2019 ◽

Author(s):

Hamad Yadikar ◽

Isabel Torres ◽

Gabrielle Aiello ◽

Milin Kurup ◽

Zhihui Yang ◽

...

Keyword(s):

Tau Protein ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Chronic Traumatic Encephalopathy ◽

Cortical Cell ◽

Protein Kinase Inhibitors ◽

Tau Hyperphosphorylation ◽

Drug Candidates ◽

A Cell ◽

Cortical Cell Cultures

ABSTRACTTauopathies are a class of neurodegenerative disorders characterized by abnormal deposition of post-translationally modified tau protein in the human brain. Tauopathies are associated with Alzheimer’s disease (AD), chronic traumatic encephalopathy (CTE), and other diseases. Hyperphosphorylation increases tau tendency to aggregate and forms neurofibrillary tangles (NFT), a pathological hallmark of AD. In this study, okadaic acid (OA, 100 nM), a protein phosphatase 1/2A inhibitor, was treated for 24h in mouse neuroblastoma (N2a) and differentiated rat primary neuronal cortical cell cultures (CTX) to induce tau-hyperphosphorylation and oligomerization as a cell-based tauopathy model. Following the treatments, the effectiveness of different kinase inhibitors was assessed using the tauopathy-relevant tau antibodies through tau-immunoblotting, including the sites: pSer202/pThr205 (AT8), pThr181 (AT270), pSer202 (CP13), pSer396/pSer404 (PHF-1), and pThr231 (RZ3). OA-treated samples induced tau phosphorylation and oligomerization at all tested epitopes, forming a monomeric band (46-67 kDa) and oligomeric bands (170 kDa and 240 kDa). We found that TBB (a casein kinase II inhibitor), AR and LiCl (GSK-3 inhibitors), cyclosporin A (calcineurin inhibitor), and Saracatinib (Fyn kinase inhibitor) caused robust inhibition of OA-induced monomeric and oligomeric p-tau in both N2a and CTX culture. Additionally, a cyclin-dependent kinase 5 inhibitor (Roscovitine) and a calcium chelator (EGTA) showed conflicting results between the two neuronal cultures.This study provides a comprehensive view of potential drug candidates (TBB, CsA, AR, and Saracatinib), and their efficacy against tau hyperphosphorylation and oligomerization processes. These findings warrant further experimentation, possibly including animal models of tauopathies, which may provide a putative Neurotherapy for AD, CTE, and other forms of tauopathy-induced neurodegenerative diseases.

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Patterns of Tyrosine Kinase Inhibitor Utilization in Newly Treated Patients With Chronic Myeloid Leukemia: An Exhaustive Population-Based Study in France

Frontiers in Oncology ◽

10.3389/fonc.2021.675609 ◽

2021 ◽

Vol 11 ◽

Author(s):

Marie Pajiep ◽

Cécile Conte ◽

Françoise Huguet ◽

Martin Gauthier ◽

Fabien Despas ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Tyrosine Kinase ◽

Myeloid Leukemia ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Antihypertensive Agents ◽

Female Ratio ◽

Increased Risk ◽

Incident Cases

We analyzed demographic characteristics, comorbidities and patterns of treatment with tyrosine kinase inhibitors (TKIs) in a cohort of 3,633 incident cases of chronic myeloid leukemia (CML) identified across France from 1 January 2011 to 31 December 2014. Patients were identified through a specific algorithm in the French Healthcare Data System and were followed up 12 months after inclusion in the cohort. The estimated incidence rate of CML for this period in France was 1.37 per 100,000 person-years (95% Confidence Interval 1.36-1.38) and was higher in men, with a peak at age 75-79 years. At baseline, the median age of the cohort was 60 years (Inter Quartile Range 47-71), the Male/Female ratio was 1.2, and 25% presented with another comorbidity. Imatinib was the first-line TKI for 77.6% of the patients, followed by nilotinib (18.3%) and dasatinib (4.1%). Twelve months after initiation, 86% of the patients remained on the same TKI, 13% switched to another TKI and 1% received subsequently three different TKIs. During the follow-up, 23% discontinued and 52% suspended the TKI. Patients received a mean of 16.7 (Standard Deviation (SD) 9.6) medications over the first year of follow-up, and a mean of 2.7 (SD 2.3) concomitant medications on the day of first TKI prescription: 24.4% of the patients received allopurinol, 6.4% proton pump inhibitors (PPI) and 6.5% antihypertensive agents. When treatment with TKI was initiated, incident CML patients presented with comorbidities and polypharmacy, which merits attention because of the persistent use of these concomitant drugs and the potential increased risk of drug-drug interactions.

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Brigatinib (BRG) in ALK+ crizotinib (CRZ)-refractory non-small cell lung cancer (NSCLC): Final results of the phase 1/2 and phase 2 (ALTA) trials.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.9071 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. 9071-9071

Author(s):

Scott N. Gettinger ◽

Rudolf M. Huber ◽

Dong-Wan Kim ◽

Lyudmila Bazhenova ◽

Karin Holmskov Hansen ◽

...

Keyword(s):

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Phase 1 ◽

Objective Response ◽

Progression Free Survival ◽

Phase 2 ◽

Open Label ◽

Long Term Follow Up

9071 Background: BRG is a kinase inhibitor approved for the treatment of patients (pts) with ALK+ metastatic NSCLC; specific details for BRG use vary by indication and country. We report long-term efficacy and safety results of the Phase 1/2 and Phase 2 (ALTA) trials of BRG. Methods: The Phase 1/2 study was a single-arm, open-label trial (NCT01449461) of BRG 30–300 mg/d in pts with advanced malignancies. ALTA (NCT02094573) randomized pts with CRZ-refractory ALK+ NSCLC to receive BRG at 90 mg qd (arm A) or 180 mg qd with 7-d lead-in at 90 mg (arm B). For the Phase 1/2 study, investigator assessments of confirmed objective response rate (cORR; RECIST v1.1), duration of response (DoR), progression-free survival (PFS), overall survival (OS), and safety in pts with ALK+ NSCLC are reported. The primary endpoint of ALTA was cORR per investigator; secondary endpoints included cORR per independent review committee (IRC), DoR, PFS, and OS. Results: In the Phase 1/2 study, 137 pts received BRG; of these, 79 pts had ALK+ NSCLC (71/79 had prior CRZ; 28/79 received 180 mg qd [7-d lead-in at 90 mg]; 14/79 received 90 mg qd). In ALTA, 222 pts with CRZ-refractory ALK+ NSCLC were randomized (n = 112/110, arm A/B). At the end of the Phase 1/2 study (Feb 18, 2020), with median 27.7 mo follow-up (̃67 mo after last pt enrolled), 4 pts remained on BRG. At the end of ALTA (Feb 27, 2020), with median 19.6/28.3 mo follow-up in arm A/B (̃53 mo after last pt enrolled), 10/17 pts in arm A/B were still on treatment. Table shows efficacy results from final analyses with long-term follow-up. In ALTA, the IRC-assessed intracranial cORR in pts with measurable baseline brain metastases was 50% (13/26) in arm A and 67% (12/18) in arm B; Kaplan-Meier (KM) estimated median intracranial DoR was 9.4 mo (95% CI, 3.7, not reached [NR]) in arm A and 16.6 mo (3.7, NR) in arm B. With long-term follow-up, no new safety signals were identified. Treatment-emergent adverse events led to dose interruption (Phase 1/2: 59%; ALTA arm A/B: 49%/61%), dose reduction (13%; 8%/33%), or discontinuation (10%; 4%/13%). Conclusions: BRG showed sustained long-term activity, PFS, and manageable safety in pts with CRZ-refractory ALK+ NSCLC. The 180 mg/d dose after 7-d lead-in at 90 mg/d led to numerically higher median PFS and OS. Final results are similar to those reported for other approved ALK tyrosine kinase inhibitors in this setting. Clinical trial information: NCT01449461, NCT02094573. [Table: see text]

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Reaction-based fluorogenic probes for selective detection of cysteinyl oxidation in living cells

10.21203/rs.3.rs-956877/v1 ◽

2021 ◽

Author(s):

Renan Ferreira ◽

Ling Fu ◽

Jing Yang ◽

Kate Carroll

Keyword(s):

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Positive Association ◽

Protein S ◽

Cell Permeability ◽

Sulfenic Acid ◽

Fluorogenic Probes ◽

A Cell ◽

Antioxidant Defense Systems ◽

S Oxidation

Abstract Measuring reactive oxygen, nitrogen and sulfur species in cells is established technology, but turn-on fluorescence tools for detecting the products of their reaction with protein cysteines remain essentially unknown. Toward this goal, here we describe fluorogenic probes for sulfenic acid, a redox modification of protein cysteines inextricably linked to signaling and oxidative stress. The probes, called CysOx1 and CysOx2, are reaction-based, exhibit excellent cell permeability, rapid reactivity, and high selectivity with minimal cytotoxicity. We applied CysOx2 in a cell-based 96-well plate assay to determine whether kinase inhibitors modulate protein S-sulfenylation as well as O-phosphorylation. Analysis of these data revealed an unexpected positive association of S-sulfenylation and inhibition of select kinases within the TK, AGC, and CMGC families including GSK3, a multitasking Ser/Thr kinase and emerging therapeutic target for neurodegenerative and mood disorders. Chemoproteomic mapping of sulfenic acid-modified cysteines in GSK3 inhibitor-treated cells shows that sites of S-oxidation localize to regulatory cysteines within key components of antioxidant defense systems. Our studies with CysOx probes offer up new insights into kinase-inhibitor dependent modulation of sulfenylome dynamics and should accelerate future efforts in the modern era of translational redox medicine.

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Optimization of Aminoimidazole Derivatives as Src Family Kinase Inhibitors

Molecules ◽

10.3390/molecules23092369 ◽

2018 ◽

Vol 23 (9) ◽

pp. 2369 ◽

Cited By ~ 1

Author(s):

Cinzia Francini ◽

Francesca Musumeci ◽

Anna Fallacara ◽

Lorenzo Botta ◽

Alessio Molinari ◽

...

Keyword(s):

Clinical Trials ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Computational Study ◽

Src Family Kinase ◽

Optimization Study ◽

New Chemical Entities ◽

A Cell ◽

Nanomolar Range ◽

Mix And Match

Protein kinases have emerged as crucial targets for cancer therapy over the last decades. Since 2001, 40 and 39 kinase inhibitors have been approved by FDA and EMA, respectively, and the majority are antineoplastic drugs. Morevoer, many candidates are currently in clinical trials. We previously reported a small library of 4-aminoimidazole and 2-aminothiazole derivatives active as Src family kinase (SFK) inhibitors. Starting from these results, we decided to perform an optimization study applying a mix and match strategy to identify a more potent generation of 4-aminoimidazoles. Firstly, a computational study has been performed, then compounds showing the best predicted docking scores were synthesized and screened in a cell-free assay for their SFK inhibitory activity. All the new chemical entities showed IC50s in the nanomolar range, with 2–130 fold increased activities compared to the previously reported inhibitors. Finally, the most active compounds have been tested on three cancer cell lines characterized by Src hyperactivation. Compounds 4k and 4l showed an interesting antiproliferative activity on SH-SY5Y neuroblastoma (NB) cell line. In this assay, the compounds resulted more potent than dasatinib, a tyrosine kinase inhibitor approved for the treatment of leukemias and in clinical trials for NB.

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Treatment with Nilotinib for Patients with Chronic Myeloid Leukemia (CML) Who Failed Prior Therapy with Imatinib and Dasatinib.

Blood ◽

10.1182/blood.v108.11.2171.2171 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2171-2171

Author(s):

Elias Jabbour ◽

Hagop Kantarjian ◽

Francis Giles ◽

Susan O’Brien ◽

Jorge Cortes

Keyword(s):

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Cross Resistance ◽

New Agents ◽

Prior Therapy ◽

Abl Kinase ◽

Imatinib Resistant ◽

Blastic Phase ◽

Complete Hematologic Response

Nilotinib (AMN107) is a selective Bcr-Abl kinase inhibitor which is aproximately 30-fold more potent than imatinib. Phase I and II studies confirmed the efficacy of nilotinib in imatinib-resistant CML. Dasatinib (BMS-354825) is a dual Src-Abl kinase inhibitor recently approved for the treatment of CML in all stages of the disease after imatinib failure. With the availability of more new tyrosine kinase inhibitors, one important question is the existence of cross-resistance between these new agents. To help answer this question, we analyzed the outcome of patients with CML who receive therapy with nilotinib after having failed therapy with dasatinib. Seven patients were treated: 6 had previously shown hematologic resistance to imatinib and one had intolerance to imatinib. Dasatinib failure was due to hematologic resistance in all 7 patients after a median of 16 weeks (range 4 to 32) on therapy. Median age was 50 years (range, 15 to 78 years); median follow-up on nilotinib was 5 months (range, 1 to 14 months). At the start of therapy with nilotinib, 1 patient was in chronic (CP), 1 in accelerated (AP), and 5 in blastic phase (BP). Mutations were found in 2 of 4 patients assessed before nilotinib therapy (F317L and E355G, respectively). All patients received nilotinib 400 mg BID (n=5) or 800 mg single daily dose (n=2). Of the 7 evaluable patients, one patient (in AP) responded (complete hematologic response). This patient had no mutations identified at baseline. At the last follow-up, 5 patients are alive: 3 with active disease, one in CHR, and one in complete molecular remission post allogeneic stem cell transplantation. We conclude that nilotinib has modest activity in patients with CML resistant to both imatinib and dasatinib.

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Late molecular recurrences in patients with chronic myeloid leukemia experiencing treatment-free remission

Blood Advances ◽

10.1182/bloodadvances.2020001772 ◽

2020 ◽

Vol 4 (13) ◽

pp. 3034-3040 ◽

Cited By ~ 3

Author(s):

Philippe Rousselot ◽

Clémence Loiseau ◽

Marc Delord ◽

Jean Michel Cayuela ◽

Marc Spentchian

Keyword(s):

Chronic Myeloid Leukemia ◽

Tyrosine Kinase ◽

Myeloid Leukemia ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Residual Disease ◽

Molecular Response ◽

Residual Rate ◽

Treatment Free Remission

Abstract Treatment-free remission (TFR) is an opportunity for patients with chronic myeloid leukemia (CML). Reported cumulative incidence curves of molecular recurrence (MRec) arbor a 2-phase shape with mainly early events, but also some late events (late MRec [LMRec]). Having discontinued our first patient in 2004, we have access to a prolonged follow-up, enabling us to characterize these late events. Over 15 years, 128 patients from our institution were registered in the Stop Imatinib (STIM; A Study for Tyrosine Kinase Inhibitors Discontinuation [A-STIM]) trial. MRec was defined by the loss of major molecular response (BCR-ABL1IS >0.1%). At the first TFR attempt, patients had been taking a tyrosine kinase inhibitor for a median of 7.1 years and in BCR-ABL1IS ≤0.01% (MR4) for a median of 4 years. The median follow-up of patients in TFR was 6.5 years. The TFR rate was estimated to be 45.6% after 7 years. For 9/65 (14%) patients experiencing MRec, recurrence occurred after 2 years in TFR (median, 3.6 years). The residual rate of MRec after 2 years was estimated to be 18%. The probability of remaining in TFR was 65.4% for patients having experienced fluctuations of their minimal residual disease (MRD) (at least 2 consecutive measurements BCR-ABL1IS >0.0032% or loss of MR4), whereas it was 100% for those with stable MRD (P = .003). After 2 years in TFR, we observed an 18% residual rate of LMRec. These late events represent 14% of all MRec and occur in patients with fluctuating MRD measurements. A long-term molecular follow-up therefore remains mandatory for CML patients in TFR. The A-STIM study was registered at www.clinicaltrials.gov as #NCT02897245.

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Is going for cure in chronic myeloid leukemia possible and justifiable?

Hematology ◽

10.1182/asheducation.v2012.1.122.3798214 ◽

2012 ◽

Vol 2012 (1) ◽

pp. 122-128 ◽

Cited By ~ 17

Author(s):

François-Xavier Mahon

Keyword(s):

Chronic Myeloid Leukemia ◽

Tyrosine Kinase ◽

Myeloid Leukemia ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Clinical Results ◽

Major Question ◽

Abl Tyrosine Kinase ◽

Show Evidence ◽

Abl Tyrosine Kinase Inhibitor

Abstract After more than a decade of treatment of chronic myeloid leukemia (CML) patients with the BCR-ABL tyrosine kinase inhibitor imatinib, and despite the impressive clinical results of this targeted therapeutic, many questions remain unresolved. One major question is how to cure CML, and the next step for the future will be to address this key issue. CML is a good model of cancer. The fact that the majority of CML patients who respond very well but discontinue tyrosine kinase inhibitors later show evidence of molecular recurrence focuses attention on the need for further research on leukemic stem cells. The challenge now is to understand why, after stopping treatment, the leukemia recurs in some patients but not in others. If we win this battle, this progress will certainly benefit the treatment and management of other leukemias and solid tumors and will validate this new topic.

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The Role of Kinase Inhibitors in the Treatment of Patients with Acute Myeloid Leukemia

American Society of Clinical Oncology Educational Book ◽

10.14694/edbook_am.2013.33.313 ◽

2013 ◽

pp. 313-318

Author(s):

Catherine C. Smith ◽

Neil P. Shah

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Clinical Success ◽

Cyclin Dependent Kinases ◽

Downstream Signaling ◽

Flt3 Inhibitors ◽

Acute Myeloid

Multiple small molecule kinase inhibitors are currently undergoing development for the treatment of acute myeloid leukemia (AML). Recently, selective and potent FLT3 inhibitors such as AC220 (quizartinib) have proven clinically effective in patients with AML with FLT3 internal tandem duplication (ITD) mutations, but inhibitors of other pathologically activated kinases in AML such as c-KIT and JAK2 have achieved less clinical success. Other classes of inhibitors currently undergoing clinical development target mediators of downstream signaling pathways such as mTOR and MEK or cell cycle machinery such as aurora kinases, PLK1, or cyclin-dependent kinases. Other than FLT3 inhibitors, most inhibitors have achieved only rare bone marrow responses, and kinase inhibitor therapy in AML remains investigational. Continuing efforts to develop kinase inhibitors for the treatment of AML will require careful selection of patients for clinical trials, translational studies to characterize responders, and investigation of combination therapy that may be capable of improving response rates and duration.

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