FLT3 kinase is a potential drug target in acute myeloid leukemia (AML). Patients with FLT3 mutations typically have higher relapse rates and worse outcomes than patients without FLT3 mutations. In this study, we investigated the suitability of various heterocycles as central cores of FLT3 inhibitors, including thieno[3,2-d]pyrimidine, pyrazolo[1,5-a]pyrimidine, imidazo[4,5-b]pyridine, pyrido[4,3-d]pyrimidine, and imidazo[1,2-b]pyridazine. Our assays revealed a series of imidazo[1,2-b]pyridazines with high potency against FLT3. Compound 34f showed nanomolar inhibitory activity against recombinant FLT3-ITD and FLT3-D835Y (IC50 values 4 and 1 nM, respectively) as well as in the FLT3-ITD-positive AML cell lines MV4-11, MOLM-13, and MOLM-13 expressing the FLT3-ITD-D835Y mutant (GI50 values of 7, 9, and 4 nM, respectively). In contrast, FLT3-independent cell lines were much less sensitive. In vitro experiments confirmed suppression of FLT3 downstream signaling pathways. Finally, the treatment of MV4-11 xenograft-bearing mice with 34f at doses of 5 and 10 mg/kg markedly blocked tumor growth without any adverse effects.
- MeSH
- akutní myeloidní leukemie * patologie MeSH
- antitumorózní látky * farmakologie terapeutické užití MeSH
- apoptóza MeSH
- inhibitory proteinkinas farmakologie terapeutické užití MeSH
- lidé MeSH
- mutace MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- pyridaziny * farmakologie terapeutické užití MeSH
- pyrimidiny farmakologie MeSH
- tyrosinkinasa 3 podobná fms genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
INTRODUCTION: Cyclin-dependent kinase 7 (CDK7) is a member of the CDK family of serine/threonine protein kinases and participates in the regulation of the cell cycle and mRNA transcription. CDK7 is emerging as a possible drug target in oncology and six exciting drug candidates have already undergone early evaluation in clinical trials. AREAS COVERED: This review examines CDK7 inhibitors as anticancer drugs reported in patents published in the online databases of the World Intellectual Property Organization and European Patent Office in the 2018-2022 period. This review provides an overview of available inhibitors, including their chemical structures, biochemical profile and stage of development. EXPERT OPINION: Small-molecule CDK7 inhibitors represent attractive pharmacological modalities for the treatment of various cancer types. Highly potent and selective inhibitors have been discovered and many of them show promising results in several preclinical cancer models. Developed compounds act on the kinase by various mechanisms, including traditional ATP competition, irreversible binding to tractable cysteine 312 outside the active site of CDK7, and induced protein degradation by proteolysis targeting chimeras. Ongoing preclinical research and clinical trials should reveal which strategy will provide the highest benefits.
- MeSH
- cyklin-dependentní kinasy genetika MeSH
- inhibitory proteinkinas farmakologie chemie MeSH
- kinasa aktivující cyklin dependentní kinasy * MeSH
- lidé MeSH
- nádory * farmakoterapie MeSH
- patenty jako téma MeSH
- protein-serin-threoninkinasy metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Therapy of FLT3-positive acute myeloid leukemia still remains complicated, despite the availability of newly approved kinase inhibitors. Various strategies to avoid the reduced efficacy of therapy have been explored, including the development of dual targeting compounds, which inhibit FLT3 and another kinase necessary for the survival and proliferation of AML cells. We have designed new 2,7,9-trisubstituted 8-oxopurines as FLT3 inhibitors and report here the structure-activity relationship studies. We demonstrated that substituents at positions 7 and 9 modulate activity between CDK4 and FLT3 kinase, and the isopropyl group at position 7 substantially increased the selectivity toward FLT3 kinase, which led to the discovery of compound 15a (9-cyclopentyl-7-isopropyl-2-((4-(piperazin-1-yl)phenyl)amino)-7,9-dihydro-8H-purin-8-one). Cellular analyses in MV4-11 cells revealed inhibition of autophosphorylation of FLT3 kinase in nanomolar doses, including the suppression of downstream STAT5 and ERK1/2 phosphorylation. We also describe mechanistic studies in cell lines and activity in a mouse xenograft model in vivo.
- MeSH
- akutní myeloidní leukemie * farmakoterapie MeSH
- apoptóza MeSH
- fosforylace MeSH
- inhibitory proteinkinas terapeutické užití MeSH
- lidé MeSH
- mutace MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- proliferace buněk MeSH
- tyrosinkinasa 3 podobná fms MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Background: Alzheimer's disease (AD) drugs in therapy are limited to acetylcholine esterase inhibitors and memantine. Newly developed drugs against a single target structure have an insufficient effect on symptomatic AD patients. Results: Novel aromatically anellated pyridofuranes have been evaluated for inhibition of AD-relevant protein kinases cdk1, cdk2, gsk-3b and Fyn. Best activities have been found for naphthopyridofuranes with a hydroxyl function as part of the 5-substituent and a hydrogen or halogen substituent in the 8-position. Best results in nanomolar ranges were found for benzopyridofuranes with a 6-hydroxy and a 3-alkoxy substitution or an exclusive 6-alkoxy substituent. Conclusion: First lead compounds were identified inhibiting two to three kinases in nanomolar ranges to be qualified as an innovative approach for AD multitargeting.
3,5,7-Trisubstituted pyrazolo[4,3-d]pyrimidines have been identified as potent inhibitors of cyclin-dependent kinases (CDKs), which are established drug targets. Herein, we describe their further structural modifications leading to novel nanomolar inhibitors with strong antiproliferative activity. We determined the crystal structure of fully active CDK2/A2 with 5-(2-amino-1-ethyl)thio-3-cyclobutyl-7-[4-(pyrazol-1-yl)benzyl]amino-1(2)H-pyrazolo[4,3-d]pyrimidine (24) at 1.7 Å resolution, confirming the competitive mode of inhibition. Biochemical and cellular assays in lymphoma cell lines confirmed the expected mechanism of action through dephosphorylation of retinoblastoma protein and RNA polymerase II, leading to induction of apoptosis. Importantly, we also revealed an interesting ability of compound 24 to induce proteasome-dependent degradation of cyclin K both in vitro and in a patient-derived xenograft in vivo. We propose that 24 has a dual mechanism of action, acting as a kinase inhibitor and as a molecular glue inducing an interaction between CDK12 and DDB1 that leads to polyubiquitination of cyclin K and its subsequent degradation.
- MeSH
- antitumorózní látky * chemie farmakologie MeSH
- cyklin-dependentní kinasa 2 MeSH
- cyklin-dependentní kinasy * MeSH
- cykliny metabolismus MeSH
- inhibitory proteinkinas chemie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- pyrimidiny chemie MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We report 31 new compounds designed, synthesized and evaluated on Bcr-Abl, BTK and FLT3-ITD as part of our program to develop 2,6,9-trisubstituted purine derivatives as inhibitors of oncogenic kinases. The design was inspired by the chemical structures of well-known kinase inhibitors and our previously developed purine derivatives. The synthesis of these purines was simple and used a microwave reactor for the final step. Kinase assays showed three inhibitors with high selectivity for each protein that were identified: 4f (IC50 = 70 nM for Bcr-Abl), 5j (IC50 = 0.41 μM for BTK) and 5b (IC50 = 0.38 μM for FLT-ITD). The 3D-QSAR analysis and molecular docking studies suggested that two fragments are potent and selective inhibitors of these three kinases: a substitution at the 6-phenylamino ring and the length and volume of the alkyl group at N-9. The N-7 and the N-methyl-piperazine moiety linked to the aminophenyl ring at C-2 are also requirements for obtaining the activity. Furthermore, most of these purine derivatives were shown to have a significant inhibitory effect in vitro on the proliferation of leukaemia and lymphoma cells (HL60, MV4-11, CEM, K562 and Ramos) at low concentrations. Finally, we show that the selected purines (4i, 5b and 5j) inhibit the downstream signalling of the respective kinases in cell models. Thus, this study provides new evidence regarding how certain chemical modifications of purine ring substituents provide novel inhibitors of target kinases as potential anti-leukaemia drugs.
- Publikační typ
- časopisecké články MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
The B-cell receptor (BCR) signalling pathway is tightly connected with the function, proliferation and survival of B cells. Aberrant BCR signalling, typical for various B cell malignancies, has been successfully targeted by small molecules kinase inhibitors that have shown an encouraging effect in clinical settings. On the other hand, adverse effects limiting the clinical use of these agents stimulate further discovery and development of drug against the BCR signalosome. This project is focused on development of novel kinase inhibitors of the BCR signalosome members: BTK (Brutton tyrosine kinase), SYK (spleen tyrosine kinase) and PI3K (phosphatidylinositol 3-kinases). We have already designed and prepared several imidazo[4,5-b]pyridines and imidazo[4,5-c]pyridines, exploited only rarely as kinase inhibitors. Within this project, the compounds will be further optimized and studied to clarify their structure-activity relationships in biochemical and cellular assays. Therapeutic efficacy of selected lead will be determined in a suitable mouse model of B-cell lymphoma.
Signalizace B-buněčného receptoru (BCR) se podílí na funkci, proliferaci a přežívání B lymfocytů. Aberantní signalizace BCR, která je typická pro různé B-buněčné malignity, je považována za racionální a poměrně efektivní cíl nízkomolekulárních sloučenin, které již prokázaly svou terapeutickou účinnost v klinických experimentech. Na druhou stranu však jejich použití omezují některé nežádoucí účinky, což nadále stimuluje k vývoji dalších inhibitorů BCR signalosomu. Tento projekt je zaměřen na vývoj nových inhibitorů kinas právě v rámci BCR signalosomu, a to konkrétně na BTK (Bruttonova tyrosinová kinasa), SYK (slezinová tyrosinová kinasa) a PI3K (fosfatidylinositol 3-kinasy). Nedávno se nám podařilo navrhnout a připravit několik imidazo[4,5-b]pyridinů a imidazo[4,5-c]pyridinů, které jako inhibitory kinas nebyly dosud využívány. V rámci tohoto projektu hodláme optimalizovat tyto sloučeniny a objasňovat vztahy mezi strukturou a aktivitou v biochemických a buněčných experimentech.Terapeutická účinnost aktivních derivátů bude případně ověřena v myším modelu vhodného B lymfomu.
- MeSH
- antitumorózní látky terapeutické užití MeSH
- B-buněčný lymfom farmakoterapie MeSH
- inhibitory fosfoinositid-3-kinasy terapeutické užití MeSH
- inhibitory tyrosinkinasy antagonisté a inhibitory MeSH
- kinasa Syk antagonisté a inhibitory MeSH
- lidé MeSH
- proteinkinasa BTK antagonisté a inhibitory MeSH
- signální transdukce MeSH
- vyvíjení léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- hodnotící studie MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- onkologie
- farmacie a farmakologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
The 3H-pyrazolo[4,3-f]quinoline moiety has been recently shown to be a privileged kinase inhibitor core with potent activities against acute myeloid leukemia (AML) cell lines in vitro. Herein, various 3H-pyrazolo[4,3-f]quinoline-containing compounds were rapidly assembled via the Doebner-Povarov multicomponent reaction from the readily available 5-aminoindazole, ketones, and heteroaromatic aldehydes in good yields. The most active compounds potently inhibit the recombinant FLT3 kinase and its mutant forms with nanomolar IC50 values. Docking studies with the FLT3 kinase showed a type I binding mode, where the 3H-pyrazolo group interacts with Cys694 in the hinge region. The compounds blocked the proliferation of AML cell lines harboring oncogenic FLT3-ITD mutations with remarkable IC50 values, which were comparable to the approved FLT3 inhibitor quizartinib. The compounds also inhibited the growth of leukemia in a mouse-disseminated AML model, and hence, the novel 3H-pyrazolo[4,3-f]quinoline-containing kinase inhibitors are potential lead compounds to develop into anticancer agents, especially for kinase-driven cancers.
- MeSH
- akutní myeloidní leukemie farmakoterapie metabolismus patologie MeSH
- antitumorózní látky chemická syntéza chemie farmakologie MeSH
- inhibitory proteinkinas chemická syntéza chemie farmakologie MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- molekulární struktura MeSH
- nádorové buňky kultivované MeSH
- proliferace buněk účinky léků MeSH
- tyrosinkinasa 3 podobná fms antagonisté a inhibitory metabolismus MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
An evaluation of possible interactions with enzymes of drug metabolism (cytochromes P450, CYP) is an important part of studies on safety and, in general, on the properties of any drug or biologically active compound. The article is focused on the preliminary metabolic study of selected 2,6,9-trisubstituted purine kinase inhibitors with significant anticancer activities which we have developed. The compounds BP-21 and BP-117 represent strong CDK inhibitors and the compound BPA-302 was developed as selective FLT3-ITD kinase inhibitor. Here, emphasis is placed on interactions of these compounds with the nine most important forms of CYP to evaluate the possibility of inhibition of these enzymes. The possibility of their inhibitory effect was studied in vitro on selected human liver microsomal CYP enzymes. The most affected enzyme was CYP2C19. Its activity dropped to 22 % of its original value by BPA 302, to 13 % by BP-21 and to 6 % by BP-117 at the highest concentration tested (250 µmol·l(-1)). The results suggest that the metabolism of concomitantly administered drugs should not be significantly affected at lower doses. Molecular docking of BPA-302 indicated that it can bind to active site of both CYP2C19 and CYP2D6 enzymes above the heme cofactor corroborating the experimental data.
- MeSH
- inhibitory cytochromu P450 chemie farmakologie MeSH
- inhibitory proteinkinas chemie farmakologie MeSH
- izoenzymy MeSH
- jaterní mikrozomy účinky léků enzymologie MeSH
- kinetika MeSH
- konformace proteinů MeSH
- lékové interakce MeSH
- lidé MeSH
- puriny chemie farmakologie MeSH
- simulace molekulového dockingu MeSH
- systém (enzymů) cytochromů P-450 chemie metabolismus MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
Treatment of acute myeloid leukemia (AML) is challenging and some AML subtypes have very poor prognosis. AML therapeutics that inhibit the oncogenic FLT3 receptor tyrosine kinase have promise but exhibit limited efficacy. However, recent findings indicate that treatments simultaneously targeting FLT3 and other rationally-selected kinases may be more efficacious. We have identified new compounds with dual specificity towards the FLT3 and CDK kinases that are structurally unrelated to other known FLT3 inhibitors. These compounds display striking efficacy and selectivity in AML cell lines with FLT3 mutations: they abrogate FLT3-dependent signaling, block proliferation and induce apoptosis in the MV4-11 cell line at low nanomolar concentrations. Other cell lines are 1000-5000x less sensitive. In this project, a series of analogous compounds will be prepared to clarify their structure-activity relationships. The most potent analogs will be characterized in biochemical and cellular assays, and their pharmacokinetic and pharmacodynamic parameters will be determined in a mouse model of AML.
Akutní myeloidní leukemie (AML) patří mezi závažná hematoonkologická onemocnění, neboť její léčba je často málo účinná. Inhibice onkogenní proteinkinasy FLT3 experimentálními inhibitory sice vykazuje klinicky lepší výsledky, ale její účinnost je také omezená. Nedávné výsledky však dokazují, že racionálně kombinovaná inhibice FLT3 a některých jiných kinas výrazně zvyšuje terapeutickou odezvu. Nedávno jsme identifikovali skupinu zcela nových sloučenin, vykazující duální specifičnost vůči kinasam FLT3 a CDK. Tyto sloučeniny se vyznačují vysokou účinností a selektivitou vůči buněčným liniím AML s mutatní FLT3; blokují FLT3-závislou buněčnou signalizaci, inhibují proliferaci a indukují apoptózu v linii MV4-11 již v nanomolárních koncentracích. Během tohoto projektu bude připravena série analogů těchto sloučenin pro objasnění vztahů mezi strukturou a aktivitou. Nejúčinnější sloučeniny budou charakterizovány v biochemických a buněčných modelech. Základní preklinické (farmakokinetické a farmakodynamické) vlastnosti budou studovány v myším modelu AML.
- MeSH
- akutní myeloidní leukemie farmakoterapie MeSH
- cyklin-dependentní kinasy MeSH
- inhibitory proteinkinas terapeutické užití MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- preklinické hodnocení léčiv MeSH
- Check Tag
- myši MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- onkologie
- biochemie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR