DOCSLIB.ORG

The Pan HDAC Inhibitor Givinostat Improves Muscle Function And

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Pan HDAC Inhibitor Givinostat Improves Muscle Function And Licandro et al. Skeletal Muscle (2021) 11:19 https://doi.org/10.1186/s13395-021-00273-6 RESEARCH Open Access The pan HDAC inhibitor Givinostat improves muscle function and histological parameters in two Duchenne muscular dystrophy murine models expressing different haplotypes of the LTBP4 gene Simonetta Andrea Licandro1,2* , Luca Crippa3, Roberta Pomarico4, Raffaella Perego4, Gianluca Fossati1, Flavio Leoni4 and Christian Steinkühler1 Abstract Background: In the search of genetic determinants of Duchenne muscular dystrophy (DMD) severity, LTBP4,a member of the latent TGF-β binding protein family, emerged as an important predictor of functional outcome trajectories in mice and humans. Nonsynonymous single-nucleotide polymorphisms in LTBP4 gene associate with prolonged ambulation in DMD patients, whereas an in-frame insertion polymorphism in the mouse LTBP4 locus modulates disease severity in mice by altering proteolytic stability of the Ltbp4 protein and release of transforming growth factor-β (TGF-β). Givinostat, a pan-histone deacetylase inhibitor currently in phase III clinical trials for DMD treatment, significantly reduces fibrosis in muscle tissue and promotes the increase of the cross-sectional area (CSA) of muscles in mdx mice. In this study, we investigated the activity of Givinostat in mdx and in D2.B10 mice, two mouse models expressing different Ltbp4 variants and developing mild or more severe disease as a function of Ltbp4 polymorphism. Methods: Givinostat and steroids were administrated for 15 weeks in both DMD murine models and their efficacy was evaluated by grip strength and run to exhaustion functional tests. Histological examinations of skeletal muscles were also performed to assess the percentage of fibrotic area and CSA increase. Results: Givinostat treatment increased maximal normalized strength to levels that were comparable to those of healthy mice in both DMD models. The effect of Givinostat in both grip strength and exhaustion tests was dose- dependent in both strains, and in D2.B10 mice, Givinostat outperformed steroids at its highest dose. The in vivo treatment with Givinostat was effective in improving muscle morphology in both mdx and D2.B10 mice by reducing fibrosis. * Correspondence: [email protected] 1New Drug Incubator, Italfarmaco S.p.A., Milan, Italy 2University of Milano-Bicocca, Milan, Italy Full list of author information is available at the end of the article © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Licandro et al. Skeletal Muscle (2021) 11:19 Page 2 of 22 Conclusion: Our study provides evidence that Givinostat has a significant effect in ameliorating both muscle function and histological parameters in mdx and D2.B10 murine models suggesting a potential benefit also for patients with a poor prognosis LTBP4 genotype. Keywords: Duchenne nuscular dystrophy, Givinostat, mdx, D2.B10, LTBP4, HDAC inhibitor Background predictor of functional outcome trajectories. The LTBP4 Duchenne muscular dystrophy (DMD) is a lethal X- gene encodes the latent transforming growth factor-β linked disorder that leads to muscle wasting with an (TGF-β)-binding protein (Ltbp) 4 that binds TGF-β in average incidence of 1:3500 newborn males. DMD is due the extracellular matrix, sequestering this cytokine [16– to mutations in the gene encoding dystrophin, which 18]. During inflammatory processes, TGF-β is released cause the absence of the protein. The lack of dystrophin from the Ltbp4 complex by proteolysis of the proline- leads to muscle fiber degeneration, activation of chronic rich hinge domain. This cleavage leads to the activation inflammatory pathways and progressive muscle tissue re- of TGF-β [19]. Once liberated from the complex, free placement by fibroblasts and adipocytes, triggering the TGF-β regulates collagen deposition and promotes fi- process leading to fibrosis development [1]. In DMD brotic processes [20]. During the physiological healing muscles, the normal regenerative pathways are subverted process, a transient release of TGF-β is necessary, and the abnormal substitution of damaged muscle fibers whereas sustained pro-inflammatory cytokine levels con- by fibrotic and adipose tissue leads to a severe reduction tribute to the pathological tissue degeneration processes, in muscle function. The main causes of the reduced life such as fibrosis [21]. High TGF-β levels have been span of DMD patients are severe respiratory insuffi- shown to correlate with the severity of muscle fibrosis in ciency, due to the weakening of the diaphragm (DIA), DMD [21]. and cardiac failure [2]. In humans, single-nucleotide polymorphisms (SNPs) To date there is no standard therapy for DMD patients have been identified in the Ltbp4 protein, which origin- that leads to the healing of the disease; however, gluco- ate VTTT and IAAM haplotypes [17]. DMD patients corticoid (GC) steroid treatment, corrective orthopedic homozygous for the IAAM Ltbp4 haplotype remained surgery, and assisted ventilation can contribute to im- ambulatory significantly longer (12.5 ± 3.3 years) than prove the quality of life of patients and to delay disease those homozygous for the VTTT haplotype (10.7 ± 2.1 progression [3]. Attempts to specifically target individual years) [17]. In addition, IAAM fibroblasts exposed to mutations have recently led to the approval of exon TGF-β show a reduction in phospho-SMAD levels when skipping oligonucleotides (Eteplirsen, Golodirsen, and compared to VTTT fibroblasts, in line with the concept Vitolarsen) by the FDA [4–6] and Ataluren for patients that LTBP4 regulates TGF-β activity [17, 22, 23]. with nonsense mutations by EMA [7, 8]. Only a minority There are two mouse strains that mimic the two dif- mdx of patients can benefit from these treatments, and the ferent human Ltbp4 haplotypes: C57BL10ScSn-Dmd / mainstay of disease management is GC steroids. GC J (hereafter referred to as mdx) have a 12-amino-acid in- steroid treatment (mainly prednisone and Deflazacort) sertion in the proline-rich region of Ltbp4. This contrib- was shown to beneficially influence all disease trajector- utes to a mild DMD phenotype due to a lower sensitivity ies, including survival and prolonged ambulation in to proteolysis and a reduced activation of TGF-β as it DMD patients [9–11], but this benefit comes at the cost occurs in the human IAAM haplotype [16]; D2.B10- mdx of significant side effects such as body weight (BW) in- Dmd /J (hereafter referred to as D2.B10) have a 12- crease, growth stunting, Cushing-like symptoms, mood amino-acid deletion in the same Ltbp4 region and func- changes, increased incidence of fractures, and suscepti- tionally resemble the human VTTT haplotype (severe bility to infections [12, 13]. Paradoxically, chronic GC DMD phenotype due to increased sensitivity to proteoly- steroid administration may promote muscle atrophy, sis and increased TGF-β activity) [16]. Mdx mice harbor and mdx mice subjected to chronic GC steroid treat- a spontaneous point mutation in exon 23 of the dys- ment show a significant impairment of heart function trophin gene, leading to the loss of dystrophin [24] and and an increase of cardiac fibrosis, suggesting that pro- are routinely used as a rodent model of the DMD dis- longed steroid treatment may be detrimental to dys- ease even though it has a milder phenotype as compared trophic heart muscle [14, 15]. Thus, there is a clear need to DMD patients and a normal lifespan. The compara- for a broadly active and well-tolerated treatment for tively mild phenotype of mdx mice can also be attributed DMD patients. to the compensatory function of the dystrophin-related Recently, in the search of genetic determinants of protein utrophin, which is highly upregulated in regener- DMD severity, the LTBP4 gene emerged as an important ating muscle fibers in adult mdx mutants, except for the Licandro et al. Skeletal Muscle (2021) 11:19 Page 3 of 22 DIA muscle [24]. The D2.B10 strain, created by back- Materials and methods crossing mdx mice onto the DBA/2J background, may Animal experiments be a superior DMD model as it better recapitulates sev- Study approval eral of the human characteristics of the DMD myopathy Procedures involving animals and their care were carried (reduced hindlimb muscle weight, fewer and atrophied out in conformity with institutional guidelines in com- myofibers, increased fibrosis, fat deposition and inflam- pliance with national and international laws and policies matory infiltrate, muscle weakness) when compared to (Italian Governing Law: D.lgs
Load more

Recommended publications
  • An Overview of the Role of Hdacs in Cancer Immunotherapy
    International Journal of Molecular Sciences Review Immunoepigenetics Combination Therapies: An Overview of the Role of HDACs in Cancer Immunotherapy Debarati Banik, Sara Moufarrij and Alejandro Villagra * Department of Biochemistry and Molecular Medicine, School of Medicine and Health Sciences, The George Washington University, 800 22nd St NW, Suite 8880, Washington, DC 20052, USA; [email protected] (D.B.); [email protected] (S.M.) * Correspondence: [email protected]; Tel.: +(202)-994-9547 Received: 22 March 2019; Accepted: 28 April 2019; Published: 7 May 2019 Abstract: Long-standing efforts to identify the multifaceted roles of histone deacetylase inhibitors (HDACis) have positioned these agents as promising drug candidates in combatting cancer, autoimmune, neurodegenerative, and infectious diseases. The same has also encouraged the evaluation of multiple HDACi candidates in preclinical studies in cancer and other diseases as well as the FDA-approval towards clinical use for specific agents. In this review, we have discussed how the efficacy of immunotherapy can be leveraged by combining it with HDACis. We have also included a brief overview of the classification of HDACis as well as their various roles in physiological and pathophysiological scenarios to target key cellular processes promoting the initiation, establishment, and progression of cancer. Given the critical role of the tumor microenvironment (TME) towards the outcome of anticancer therapies, we have also discussed the effect of HDACis on different components of the TME. We then have gradually progressed into examples of specific pan-HDACis, class I HDACi, and selective HDACis that either have been incorporated into clinical trials or show promising preclinical effects for future consideration.
    [Show full text]
  • Vorinostat—An Overview Aditya Kumar Bubna
    E-IJD RESIDENTS' PAGE Vorinostat—An Overview Aditya Kumar Bubna Abstract From the Consultant Vorinostat is a new drug used in the management of cutaneous T cell lymphoma when the Dermatologist, Kedar Hospital, disease persists, gets worse or comes back during or after treatment with other medicines. It is Chennai, Tamil Nadu, India an efficacious and well tolerated drug and has been considered a novel drug in the treatment of this condition. Currently apart from cutaneous T cell lymphoma the role of Vorinostat for Address for correspondence: other types of cancers is being investigated both as mono-therapy and combination therapy. Dr. Aditya Kumar Bubna, Kedar Hospital, Mugalivakkam Key Words: Cutaneous T cell lymphoma, histone deacytelase inhibitor, Vorinostat Main Road, Porur, Chennai - 600 125, Tamil Nadu, India. E-mail: [email protected] What was known? • Vorinostat is a histone deacetylase inhibitor. • It is an FDA approved drug for the treatment of cutaneous T cell lymphoma. Introduction of Vorinostat is approximately 9. Vorinostat is slightly Vorinostat is a histone deacetylase (HDAC) inhibitor, soluble in water, alcohol, isopropanol and acetone and is structurally belonging to the hydroxymate group. Other completely soluble in dimethyl sulfoxide. drugs in this group include Givinostat, Abexinostat, Mechanism of action Panobinostat, Belinostat and Trichostatin A. These Vorinostat is a broad inhibitor of HDAC activity and inhibits are an emergency class of drugs with potential anti- class I and class II HDAC enzymes.[2,3] However, Vorinostat neoplastic activity. These drugs were developed with the does not inhibit HDACs belonging to class III. Based on realization that apart from genetic mutation, alteration crystallographic studies, it has been seen that Vorinostat of HDAC enzymes affected the phenotypic and genotypic binds to the zinc atom of the catalytic site of the HDAC expression in cells, which in turn lead to disturbed enzyme with the phenyl ring of Vorinostat projecting out of homeostasis and neoplastic growth.
    [Show full text]
  • Evaluation of the Therapeutic Potential of the Novel Isotype Specific HDAC Inhibitor 4SC-202 in Urothelial Carcinoma Cell Lines
    Targ Oncol DOI 10.1007/s11523-016-0444-7 ORIGINAL RESEARCH ARTICLE Evaluation of the Therapeutic Potential of the Novel Isotype Specific HDAC Inhibitor 4SC-202 in Urothelial Carcinoma Cell Lines Maria Pinkerneil1 & Michèle J. Hoffmann1 & Hella Kohlhof2 & Wolfgang A. Schulz1 & Günter Niegisch1 # The Author(s) 2016. This article is published with open access at Springerlink.com Abstract Results 4SC-202 significantly reduced proliferation of all ep- Background Targeting of class I histone deacetylases ithelial and mesenchymal UC cell lines (IC50 0.15–0.51 μM), (HDACs) exerts antineoplastic actions in various cancer types inhibited clonogenic growth and induced caspase activity. by modulation of transcription, upregulation of tumor sup- Flow cytometry revealed increased G2/M and subG1 fractions pressors, induction of cell cycle arrest, replication stress and in VM-CUB1 and UM-UC-3 cells. Both effects were stronger promotion of apoptosis. Class I HDACs are often deregulated than with SAHA treatment. in urothelial cancer. 4SC-202, a novel oral benzamide type Conclusion Specific pharmacological inhibition of class I HDAC inhibitor (HDACi) specific for class I HDACs HDACs by 4SC-202 impairs UC cell viability, inducing cell HDAC1, HDAC2 and HDAC3 and the histone demethylase cycle disturbances and cell death. Combined inhibition of LSD1, shows substantial anti-tumor activity in a broad range HDAC1, HDAC2 and HDAC3 seems to be a promising treat- of cancer cell lines and xenograft tumor models. ment strategy for UC. Aim The aim of this study was to investigate the therapeutic potential of 4SC-202 in urothelial carcinoma (UC) cell lines. Methods We determined dose response curves of 4SC-202 by KeyPoints MTT assay in seven UC cell lines with distinct HDAC1, 4SC-202 exerts significant antineoplastic effects on HDAC2 and HDAC3 expression profiles.
    [Show full text]
  • Effect of Givinostat, an HDAC Inhibitor, on Disease Milestones in Duchenne Muscular Dystrophy Boys Paolo Bettica1, M.D., Ph.D., Giacomo P
    Effect of Givinostat, an HDAC inhibitor, on disease milestones in Duchenne Muscular Dystrophy boys Paolo Bettica1, M.D., Ph.D., Giacomo P. Comi2, M.D., Enrico Bertini3, M.D., Giuseppe Vita3, M.D., Eugenio Mercuri4, M.D, Sara Cazzaniga1§, M.Sc. 1 Italfarmaco S.p.A., Italy; 2 Dino Ferrari Centre Foundation IRCCS Ca’ Granda Ospedale Maggiore Policlinico, University of Milan, Italy; 3 Bambino Gesù Children's Hospital, IRCCS, Rome. Italy; 3 University of Messina, NEMO Clinical Centre, Messina, Italy; 4 Catholic University, Rome, Italy; Corresponding Author§ email: [email protected] PHASE 3 TRIAL Phase 3, multicentre, double blind, placebo controlled (2:1) study in 242 patients to What happens at study visits? • demonstrate that Givinostat oral suspension preserves muscle mass and slows down disease Informed Consent Paperwork • progression. The study is ongoing in USA, Canada and European countries. A total of 15 visits (every 3 months): • Blood draw more frequently during the first 3 months: • first month: weekly • second month: every 2 weeks • from the third month: every 3 months What does participant entail?: • Surveys (baseline, at 12 and 18 months) and Diaries • must be ambulant DMD boys from 6 years (every visit) of age, • Muscle tests every 3 months (6MWT, NSAA, 4SC, QMT) • on stable corticosteroid for at least 6 • Pulmonary Function test baseline, at 12 and 18 months months prior to start the treatment, • Thigh muscle MRI: baseline, at 12 and 18 months • able to perform the 4 stairs climb in no • Upon successful completion of the study, participants, more than 8 seconds and time to stand up regardless the ability to walk, will have the opportunity to in ≥ 3 and less than 10 seconds, enter into long term safety study and they will ALL receive the • do the MRI scan drug Givinostat Mechanism of Action in Duchenne Downstream effects of the Impact on the lack of dystrophin epigenetic effects of the lack of dystrophin Mechanical effects : .
    [Show full text]
  • Safety and Efficacy of the Maximum Tolerated Dose of Givinostat in Polycythemia Vera
    Leukemia (2020) 34:2234–2237 https://doi.org/10.1038/s41375-020-0735-y LETTER Chronic myeloproliferative neoplasms Safety and efficacy of the maximum tolerated dose of givinostat in polycythemia vera: a two-part Phase Ib/II study 1 2 3 4 5,6 Alessandro Rambaldi ● Alessandra Iurlo ● Alessandro M. Vannucchi ● Richard Noble ● Nikolas von Bubnoff ● 7 8 9 10 11 12 Attilio Guarini ● Bruno Martino ● Antonio Pezzutto ● Giuseppe Carli ● Marianna De Muro ● Stefania Luciani ● 13 14 15 16 17 Mary Frances McMullin ● Nathalie Cambier ● Jean-Pierre Marolleau ● Ruben A. Mesa ● Raoul Tibes ● 3 3 18 18 18 Alessandro Pancrazzi ● Francesca Gesullo ● Paolo Bettica ● Sara Manzoni ● Silvia Di Tollo Received: 15 October 2019 / Revised: 6 December 2019 / Accepted: 29 January 2020 / Published online: 11 February 2020 © The Author(s) 2020. This article is published with open access To the Editor: Recently, ropeginterferon α-2b was approved by European Medicinal Agency as first line for patients without symp- Polycythemia vera (PV) is a chronic myeloproliferative tomatic splenomegaly [3]. Ruxolitinib is second-line for neoplasm (cMPN) characterized by stem cell-derived clonal patients who are refractory and/or intolerant to hydroxyurea myeloproliferation resulting in panmyelosis with persis- [4]; other treatments include busulfan, pipobroman [5], and 1234567890();,: 1234567890();,: tently raised hematocrit, increased risk of thrombotic com- nonpegylated and pegylated interferons (off-label) [1, 6, 7], plications, and predisposition to evolve to myelofibrosis or but use is limited by side effects and safety concerns. leukemia [1]. Therapy is currently based on phlebotomy to Additional, targeted therapies are therefore needed. normalize hematocrit, and aspirin. Hydroxyurea is used as Up to 98% of patients with PV bear the JAK2V617F gene first line when cytoreduction is necessary [1], although mutation, which activates erythropoietin receptor signaling toxicity can result in inadequate disease management [2].
    [Show full text]
  • Romidepsin Enhances the Efficacy of Cytarabine in Vivo, Revealing Histone Deacetylase Inhibition As a Promising Therapeutic Stra
    LETTERS TO THE EDITOR treated with high-dose cytarabine developed severe Romidepsin enhances the efficacy of cytarabine myelosuppression in comparison to the other cohorts in vivo, revealing histone deacetylase inhibition as a (Figure 1B). In particular, there was a statistically signifi- promising therapeutic strategy for cant reduction in mean hemoglobin (98 vs. 42.5 g/L; KMT2A-rearranged infant acute lymphoblastic P<0.0001), white blood cell (2.43 vs. 0.13x109/L; leukemia P<0.0001) and platelet (757 vs. 294x109/L; P<0.0021) count between the mice treated with romidepsin and low- Acute lymphoblastic leukemia (ALL) in infants diag- dose cytarabine combination therapy compared to those nosed at less than 12 months of age is an aggressive malig- treated with high-dose cytarabine. nancy with a poor prognosis. Rearrangements of the Three xenograft models, PER-785, MLL-5 and MLL-14, KMT2A gene (KMT2A-r) are present in up to 80% of were used to determine the response to drug treatment by 1 cases, with 5-year event-free survival (EFS) less than 40%. EFS. MLL-5 and MLL-14 are well characterized patient- Dose intensive chemotherapy has been incorporated into derived xenografts which harbor t(10;11) and t(11;19) contemporary treatment regimens; however, this has translocations respectively.5 MLL-5 and MLL-14 were increased the burden of toxicity during therapy and late selected to test whether findings could be validated in 1,2 effects in survivors. There is a desperate need to identify independent models with distinct translocation partners. novel therapies to improve outcome.
    [Show full text]
  • Histone Deacetylase Inhibitors: a Prospect in Drug Discovery Histon Deasetilaz İnhibitörleri: İlaç Keşfinde Bir Aday
    Turk J Pharm Sci 2019;16(1):101-114 DOI: 10.4274/tjps.75047 REVIEW Histone Deacetylase Inhibitors: A Prospect in Drug Discovery Histon Deasetilaz İnhibitörleri: İlaç Keşfinde Bir Aday Rakesh YADAV*, Pooja MISHRA, Divya YADAV Banasthali University, Faculty of Pharmacy, Department of Pharmacy, Banasthali, India ABSTRACT Cancer is a provocative issue across the globe and treatment of uncontrolled cell growth follows a deep investigation in the field of drug discovery. Therefore, there is a crucial requirement for discovering an ingenious medicinally active agent that can amend idle drug targets. Increasing pragmatic evidence implies that histone deacetylases (HDACs) are trapped during cancer progression, which increases deacetylation and triggers changes in malignancy. They provide a ground-breaking scaffold and an attainable key for investigating chemical entity pertinent to HDAC biology as a therapeutic target in the drug discovery context. Due to gene expression, an impending requirement to prudently transfer cytotoxicity to cancerous cells, HDAC inhibitors may be developed as anticancer agents. The present review focuses on the basics of HDAC enzymes, their inhibitors, and therapeutic outcomes. Key words: Histone deacetylase inhibitors, apoptosis, multitherapeutic approach, cancer ÖZ Kanser tedavisi tüm toplum için büyük bir kışkırtıcıdır ve ilaç keşfi alanında bir araştırma hattını izlemektedir. Bu nedenle, işlemeyen ilaç hedeflerini iyileştirme yeterliliğine sahip, tıbbi aktif bir ajan keşfetmek için hayati bir gereklilik vardır. Artan pragmatik kanıtlar, histon deasetilazların (HDAC) kanserin ilerleme aşamasında deasetilasyonu arttırarak ve malignite değişikliklerini tetikleyerek kapana kısıldığını ifade etmektedir. HDAC inhibitörleri, ilaç keşfi bağlamında terapötik bir hedef olarak HDAC biyolojisiyle ilgili kimyasal varlığı araştırmak için, çığır açıcı iskele ve ulaşılabilir bir anahtar sağlarlar.
    [Show full text]
  • Patent Application Publication ( 10 ) Pub . No . : US 2019 / 0192440 A1
    US 20190192440A1 (19 ) United States (12 ) Patent Application Publication ( 10) Pub . No. : US 2019 /0192440 A1 LI (43 ) Pub . Date : Jun . 27 , 2019 ( 54 ) ORAL DRUG DOSAGE FORM COMPRISING Publication Classification DRUG IN THE FORM OF NANOPARTICLES (51 ) Int . CI. A61K 9 / 20 (2006 .01 ) ( 71 ) Applicant: Triastek , Inc. , Nanjing ( CN ) A61K 9 /00 ( 2006 . 01) A61K 31/ 192 ( 2006 .01 ) (72 ) Inventor : Xiaoling LI , Dublin , CA (US ) A61K 9 / 24 ( 2006 .01 ) ( 52 ) U . S . CI. ( 21 ) Appl. No. : 16 /289 ,499 CPC . .. .. A61K 9 /2031 (2013 . 01 ) ; A61K 9 /0065 ( 22 ) Filed : Feb . 28 , 2019 (2013 .01 ) ; A61K 9 / 209 ( 2013 .01 ) ; A61K 9 /2027 ( 2013 .01 ) ; A61K 31/ 192 ( 2013. 01 ) ; Related U . S . Application Data A61K 9 /2072 ( 2013 .01 ) (63 ) Continuation of application No. 16 /028 ,305 , filed on Jul. 5 , 2018 , now Pat . No . 10 , 258 ,575 , which is a (57 ) ABSTRACT continuation of application No . 15 / 173 ,596 , filed on The present disclosure provides a stable solid pharmaceuti Jun . 3 , 2016 . cal dosage form for oral administration . The dosage form (60 ) Provisional application No . 62 /313 ,092 , filed on Mar. includes a substrate that forms at least one compartment and 24 , 2016 , provisional application No . 62 / 296 , 087 , a drug content loaded into the compartment. The dosage filed on Feb . 17 , 2016 , provisional application No . form is so designed that the active pharmaceutical ingredient 62 / 170, 645 , filed on Jun . 3 , 2015 . of the drug content is released in a controlled manner. Patent Application Publication Jun . 27 , 2019 Sheet 1 of 20 US 2019 /0192440 A1 FIG .
    [Show full text]
  • Histone Deacetylase Inhibitors As Anticancer Drugs
    International Journal of Molecular Sciences Review Histone Deacetylase Inhibitors as Anticancer Drugs Tomas Eckschlager 1,*, Johana Plch 1, Marie Stiborova 2 and Jan Hrabeta 1 1 Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University and University Hospital Motol, V Uvalu 84/1, Prague 5 CZ-150 06, Czech Republic; [email protected] (J.P.); [email protected] (J.H.) 2 Department of Biochemistry, Faculty of Science, Charles University, Albertov 2030/8, Prague 2 CZ-128 43, Czech Republic; [email protected] * Correspondence: [email protected]; Tel.: +42-060-636-4730 Received: 14 May 2017; Accepted: 27 June 2017; Published: 1 July 2017 Abstract: Carcinogenesis cannot be explained only by genetic alterations, but also involves epigenetic processes. Modification of histones by acetylation plays a key role in epigenetic regulation of gene expression and is controlled by the balance between histone deacetylases (HDAC) and histone acetyltransferases (HAT). HDAC inhibitors induce cancer cell cycle arrest, differentiation and cell death, reduce angiogenesis and modulate immune response. Mechanisms of anticancer effects of HDAC inhibitors are not uniform; they may be different and depend on the cancer type, HDAC inhibitors, doses, etc. HDAC inhibitors seem to be promising anti-cancer drugs particularly in the combination with other anti-cancer drugs and/or radiotherapy. HDAC inhibitors vorinostat, romidepsin and belinostat have been approved for some T-cell lymphoma and panobinostat for multiple myeloma. Other HDAC inhibitors are in clinical trials for the treatment of hematological and solid malignancies. The results of such studies are promising but further larger studies are needed.
    [Show full text]
  • Improving Drug Discovery Using Image-Based Multiparametric Analysis of Epigenetic Landscape
    bioRxiv preprint doi: https://doi.org/10.1101/541151; this version posted February 5, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Improving drug discovery using image-based multiparametric analysis of epigenetic landscape. Chen Farhy1, Luis Orozco1, Fu-Yue Zeng1, Ian Pass1, Jarkko Ylanko2, Santosh Hariharan2, Chun-Teng Huang1, David Andrews2,3, and Alexey V. Terskikh1*. 1Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, USA 2Biological Sciences Platform, Sunnybrook Research Institute. 3Departments of Biochemistry and Medical Biophysics, University of Toronto, Ontario, Canada. *Correspondence should be addressed to A.V.T. ([email protected]). Abstract With the advent of automatic cell imaging and machine learning, high-content phenotypic screening has become the approach of choice for drug discovery due to its ability to extract drug specific multi- layered data and compare it to known profiles. In the field of epigenetics such screening approaches has suffered from the lack of tools sensitive to selective epigenetic perturbations. Here we describe a novel approach Microscopic Imaging of Epigenetic Landscapes (MIEL) that captures patterns of nuclear staining of epigenetic marks (e.g. acetylated and methylated histones) and employs machine learning to accurately distinguish between such patterns (1). We demonstrated that MIEL has superior resolution compared to conventional intensity thresholding techniques and enables efficient detection of epigenetically active compounds, function-based classification, flagging possible off-target effects and even predict novel drug function. We validated MIEL platform across multiple cells lines and using dose-response curves to insure the robustness of this approach for the high content high throughput drug discovery.
    [Show full text]
  • Downregulation of Cell Cycle and Checkpoint Genes by Class I HDAC Inhibitors Limits Synergism with G2/M Checkpoint Inhibitor MK-1775 in Bladder Cancer Cells
    G C A T T A C G G C A T genes Article Downregulation of Cell Cycle and Checkpoint Genes by Class I HDAC Inhibitors Limits Synergism with G2/M Checkpoint Inhibitor MK-1775 in Bladder Cancer Cells Michèle J. Hoffmann * , Sarah Meneceur †, Katrin Hommel †, Wolfgang A. Schulz and Günter Niegisch Department of Urology, Medical Faculty, Heinrich-Heine-University Duesseldorf, Moorenstr. 5, 40225 Duesseldorf, Germany; [email protected] (S.M.); [email protected] (K.H.); [email protected] (W.A.S.); [email protected] (G.N.) * Correspondence: [email protected]; Tel.: +49-21-1811-5847 † Contributed equally. Abstract: Since genes encoding epigenetic regulators are often mutated or deregulated in urothelial carcinoma (UC), they represent promising therapeutic targets. Specifically, inhibition of Class-I histone deacetylase (HDAC) isoenzymes induces cell death in UC cell lines (UCC) and, in contrast to other cancer types, cell cycle arrest in G2/M. Here, we investigated whether mutations in cell cycle genes contribute to G2/M rather than G1 arrest, identified the precise point of arrest and clarified the function of individual HDAC Class-I isoenzymes. Database analyses of UC tissues and cell lines revealed mutations in G1/S, but not G2/M checkpoint regulators. Using class I-specific HDAC inhibitors (HDACi) with different isoenzyme specificity (Romidepsin, Entinostat, RGFP966), cell cycle arrest was shown to occur at the G2/M transition and to depend on inhibition of HDAC1/2 Citation: Hoffmann, M.J.; rather than HDAC3. Since HDAC1/2 inhibition caused cell-type-specific downregulation of genes Meneceur, S.; Hommel, K.; Schulz, encoding G2/M regulators, the WEE1 inhibitor MK-1775 could not overcome G2/M checkpoint W.A.; Niegisch, G.
    [Show full text]
  • Mechanism of Action of Histone Deacetylase Inhibitor - Givinostat in Chronic Myeloproliferative Neoplasm
    PhD Program in Translational and Molecular Medicine DIMET (XXV cycle, academic year 2010/2012) University of Milano-Bicocca School of Medicine and Faculty of Science Mechanism of action of Histone Deacetylase Inhibitor - Givinostat in Chronic Myeloproliferative Neoplasm 2 Table of Contents Chapter 1: Introduction 5 1.1 Chromatin Structure and Remodeling 6 1.1.1 Chromatin Structure 6 1.1.2 Epigenetic and Histone Modifying Enzymes 8 1.1.2.1 Histone Acetyltransferase 11 1.1.2.2 Histone Deacetylase 16 1.1.3 Chromatin Modification in Cancer and Histone 20 1.1.3.1 Histone Deacetylation Inhibitor 25 1.1.3.2 Histone Deacetylation Inhibitor Classes 28 1.1.3.3 Anti-Tumor Activities of Histone Deacetylation Inhibitor 33 1.2 Description of Givinostat 44 1.2.1 In vitro activity of GVS against hematologic tumors 45 1.2.2 Toxicology Studies on GVS 50 1.3 Classic Myeloproliferative Neoplasm 51 1.3.1 Normal Hematopoiesis 51 1.3.2 Myeloproliferative Neoplasm (MPN) 55 1.3.3 Polycytemia Vera (PV) 59 1.3.4 Essential Thrombocytosis (ET) 61 1.3.5 Primary Myelofibrosis (PMF) 62 1.4 Molecular Pathogenesis in Classic MPN 64 1.4.1 JAK2-STAT5 pathway 64 1.4.2 JAK2V617F mutation in MPN 67 3 1.5 Scope of the Thesis 73 1.6 Reference 74 Chapter 2: The HDAC inhibitor Givinostat modulates the hematopoietic 84 transcription factors NF-E2 and C-MYB in JAK2(V617F) myeloproliferative neoplasm cells. Exp Hematol. 2012 Aug;40(8):634-45. Chapter 3: Givinostat and Hydroxyurea synergize in vitro to induce 130 apoptosis of cells from JAK2V617F Myeloproliferative Neoplasm patients.
    [Show full text]