US20150352065A1 - Methods of treatment of fibrosis and cancers - Google Patents

Methods of treatment of fibrosis and cancers Download PDF

Info

Publication number
US20150352065A1
US20150352065A1 US14/760,880 US201414760880A US2015352065A1 US 20150352065 A1 US20150352065 A1 US 20150352065A1 US 201414760880 A US201414760880 A US 201414760880A US 2015352065 A1 US2015352065 A1 US 2015352065A1
Authority
US
United States
Prior art keywords
compound
fibrosis
cancer
formula
treatment
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US14/760,880
Other languages
English (en)
Inventor
Rémy Hanf
Dean Hum
Robert Walczak
Benoît NOEL
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Genfit SA
Original Assignee
Genfit SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Genfit SA filed Critical Genfit SA
Publication of US20150352065A1 publication Critical patent/US20150352065A1/en
Assigned to GENFIT reassignment GENFIT ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HANF, REMY, NOEL, BENOIT, WALCZAK, ROBERT, HUM, DEAN
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • the present invention relates to the use of compound 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one for treating fibrotic diseases and cancers.
  • Tyrosine kinases are important mediators of the signaling cascade, determining key roles in diverse biological processes like growth, differentiation, metabolism and apoptosis in response to external and internal stimuli. Recent advances have implicated the role of tyrosine kinases in the pathophysiology of fibrosis or cancers.
  • tyrosine kinases have been identified as determinants of disease progression and potential targets for anti-fibrotic therapies. This includes both receptor tyrosine kinases (e.g., PDGF receptor, VEGF receptor, EGF receptor, and JAK kinases) as well as non-receptor tyrosine kinases (e.g., c-Abl, c-Kit, and Src kinases) [1].
  • receptor tyrosine kinases e.g., PDGF receptor, VEGF receptor, EGF receptor, and JAK kinases
  • non-receptor tyrosine kinases e.g., c-Abl, c-Kit, and Src kinases
  • PDGFs are the primary mitogens for cells of mesenchymal origin. Excessive activity of PDGF has been associated with several human disorders, including organ fibrosis and tumorigenesis [3]. Elevated PDGF levels or activity has in particular been reported in pulmonary fibrosis, liver fibrosis [2], scleroderma [4-6], renal fibroproliferative diseases [7, 8], myeloproliferative diseases such as idiopathic myelofibrosis [9, 10], leukemia, in particular in leukemic cells of the bone marrow during progression of acute megakaryoblastic leukemia manifesting myelofibrosis [11] and chronic myelogenous leukemia [12-14].
  • VEGFRs can cause over-activation of the Raf/MEK/ERK mitogen-activated protein (MAP) kinase signaling pathway [18, 19].
  • MAP Raf/MEK/ERK mitogen-activated protein
  • Activation of the Raf/MEK/ERK mitogen-activated protein (MAP) kinase signaling pathway is known to increase cell proliferation and survival directly, and can indirectly stimulate angiogenesis by increasing the production of VEGF and PDGF [18].
  • HCC hepatocellular carcinoma
  • Sorafenib inhibits the upstream receptor tyrosine kinases that are important in angiogenesis, including VEGFR-2, VEGFR-3, PDGFR ⁇ , and kit and Raf serine/threonine kinase isoforms (e.g. Raf-1 and B-Raf).
  • Sorafenib can induce tumor cell death and inhibit angiogenesis.
  • Sorafenib has also been shown to induce apoptosis in several tumor cell lines through mechanisms that are not well established [20, 21]. Sorafenib is the first FDA-approved systemic therapy for patients with advanced HCC not amenable to treatment by surgical resection or liver transplantation.
  • the present invention provides novel uses of 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one in the preparation of pharmaceutical compositions for treating fibrotic diseases and cancers.
  • 1-[4-Methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one has anti-fibrotic and anti-cancer properties through inhibition of receptor and non-receptor tyrosine kinases and/or inhibition of their intracellular signaling pathways in fibroblasts.
  • Col1a1 collagen, type I, alpha 1
  • Col4a1 collagen, type 4, alpha 1
  • EGFR Epidermal Growth Factor Receptor
  • ERK Extracellular signal Regulated Kinase
  • FBS fetal bovine serum
  • FCS fetal calf serum
  • FGFR Fibroblast Growth Factor Receptor
  • HCC Hepatocellular Carcinoma
  • HGFR Hepatocyte Growth Factor Receptor
  • hHSC human primary hepatic stellate cells
  • MEK Mitogen-Activated Protein Kinase Kinase
  • PDGFR Platelet-Derived Growth Factor Receptor
  • RCC Renal Cell Carcinoma
  • TGF ⁇ Transforming Growth Factor beta Receptor
  • VEGFR Vascular Endothelial Growth Factor Receptor
  • FIG. 1 Compound of formula (I) interferes with hHSC cell proliferation that was induced by PDGF-BB
  • hHSC human primary hepatic stellate cells
  • PDGF-BB 10 ng/mL
  • Crenolanib a potent PDGFR inhibitor was used as a positive control. Crenolanib and the compound of formula (I) were added to the cell culture 1 hour before the stimulation with PDGF-BB in a serum free medium. Cell proliferation was assessed after 24 hours of incubation by measuring the incorporation of bromodeoxyuridine (BrdU).
  • FIG. 2 Compound of formula (I) interferes with hHSC cell proliferation that was induced by serum
  • hHSC human primary hepatic stellate cells
  • FBS fetal bovine serum
  • Crenolanib a potent PDGFR inhibitor was used as a positive control. Crenolanib and compound of formula (I) were added to the cell culture 1 hour before the stimulation with FBS. Cell proliferation was assessed after 48 hours of incubation by measuring the incorporation of bromodeoxyuridine (BrdU).
  • FIG. 3 Compound of formula (I) interferes with PDGF-BB induced PDGFRI3 phosphorylation
  • the phosphorylation of PDGFR ⁇ was induced in hHSCs with PDGF-BB, in serum free conditions.
  • the HSCs were first treated for 60 minutes with either Crenolanib (PDGFR inhibitor) or with compound of formula (I), then incubated for 10 minutes with PDGF-BB (30 ng/mL).
  • the extent of the PDGFR ⁇ phosphorylation on the tyrosine 751 was then determined by using the Human Phospho-PDGFR ⁇ (Y751) Cell-Based ELISA kit (R&D Systems), upon fixation in culture wells.
  • FIG. 4 Treatment with compound of formula (I) prevented the induction of colonic fibrosis in a model of inflammatory bowel disease.
  • ⁇ -SMA a recognized biomarker of fibrotic response, in TNBS-induced colitis was partially prevented by the administration of compound of formula (I).
  • FIG. 5 Treatment with compound of formula (I) induced a modest proapoptotic response in activated hHSCs and significantly potentiated the proapoptotic effect of staurosporine
  • the present invention relates to the use of compound 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one for treating fibrosis and cancers, such compound being capable of decreasing in an unexpected manner proliferation and activation of human fibroblasts including stellate cells, the main cellular type responsible for formation of extracellular matrix and fibrosis.
  • Compound 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyl-oxyphenyl]prop-2-en-1-one to be used according to the invention has the following Formula (I):
  • the invention relates to compound 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one for use in a method for the treatment or prevention of fibrotic diseases, wherein the fibrotic disease is not liver fibrosis, or for the treatment or prevention of a tyrosine kinase related cancer.
  • the invention relates to the compound of Formula (I) for use in the inhibition of proliferation and/or activation of cells responsible for the production of collagen fibers and/or responsible for the production of the extracellular matrix.
  • the invention further relates to the compound of Formula (I) for use in promoting apoptosis of cells responsible for the production of collagen fibers and/or responsible for the production of the extracellular matrix.
  • fibrosis includes in particular a lung, heart, muscle, skin, soft tissue (e.g. mediastinum or retroperitoneum), bone marrow, intestinal, and joint (e.g. knee, shoulder or other joints) fibrosis.
  • fibrosis includes, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endomyocardial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis (a complication of coal workers' pneumoconiosis), nephrogenic systemic fibrosis, Crohn's disease, keloid, old myocardial infarction, scleroderma/systemic sclerosis, arthrofibrosis and some forms of adhesive capsulitis.
  • fibrosis does not include liver fibrosis in the context of the present invention.
  • Receptor tyrosine kinases are widely abundant and dysregulated in cancers, and have been the focus of targeted therapies for several decades, using small molecules or antibodies [22-24].
  • the wording “tyrosine kinase-related cancer” means any form of cancer that relies on a deregulated activity or expression of a single or a group of tyrosine kinase receptors.
  • the tyrosine kinase is a receptor tyrosine kinase, more particularly PDGFR, VEGFR, FGFR, EGFR, c-Kit, or JAK kinases or non-receptor tyrosine kinases, more particularly c-Abl, or Src kinases.
  • cancer includes hepatocellular carcinoma, renal cell carcinoma, gastrointestinal stromal tumor (GIST), gastric cancer, menigioma associated with neurofibromatosis, pancreatic neuroendocrine tumors, pancreatic exocrine tumors, leukemia, myeloproliferative/myelodisplastic diseases, mastocytosis, dermatofibrosarcoma, solid tumors including breast, lung, thyroid and colorectal cancers, prostate cancer
  • GIST gastrointestinal stromal tumor
  • the invention relates to the curative treatment of a liver cancer, in particular of a hepatocellular carcinoma.
  • the invention relates to the treatment or the prevention of a cancer different from a liver cancer.
  • the cancer may be selected from renal cell carcinoma, gastrointestinal stromal tumor (GIST), menigioma associated with neurofibromatosis, pancreatic neuroendocrine tumors, pancreatic exocrine tumors leukemia, myeloproliferative/myelodisplastic diseases, mastocytosis, dermatofibrosarcoma, solid tumors including breast, lung, thyroid and colorectal cancers, prostate cancer.
  • the prevented or treated cancer is a fibrotic cancer.
  • the treatment or prevention involves the administration of the compound or a pharmaceutical composition containing the same to a patient having a declared disorder to cure, delay, or slow down the progress, thus improving the condition of the patient or to a healthy subject, in particular a subject who is at risk of developing a fibrotic disease.
  • the subjects to be treated according to the invention can be selected on the basis of several criteria associated to fibrotic diseases or the cancers such as previous drug treatments, associated pathologies, genotype, exposure to risk factors, viral infection, as well as any other relevant biomarker that can be evaluated by means of imaging methods and immunological, biochemical, enzymatic, chemical, or nucleic acid detection method.
  • Synthesis of compound 1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one can for example be carried out as described for compound 29 in WO2004/005233.
  • the compound of Formula (I) can be formulated as pharmaceutically acceptable salts, being slightly- or non-toxic salts obtained from organic or inorganic bases or acids of compound of Formula (I). These salts can be obtained during the final purification step of the compound or by incorporating the salt into the previously purified compound.
  • compositions comprising a compound of Formula (I) for the treatment of fibrotic diseases or cancers can comprise one or several excipients or vehicles, acceptable within a pharmaceutical context (e.g. saline solutions, physiological solutions, isotonic solutions, etc., compatible with pharmaceutical usage and well-known by one of ordinary skill in the art).
  • These compositions can comprise one or several agents or vehicles chosen among dispersants, solubilisers, stabilisers, preservatives, etc.
  • Agents or vehicles useful for these formulations are particularly methylcellulose, hydroxymethylcellulose, carboxymethylcellulose, polysorbate 80, mannitol, gelatin, lactose, vegetable oils, acacia, liposomes, etc.
  • compositions can be formulated in the form of injectable suspensions, gels, oils, pills, suppositories, powders, gel caps, capsules, aerosols, etc., eventually by means of galenic forms or devices assuring a prolonged and/or slow release.
  • agents such as cellulose, carbonates or starches can advantageously be used.
  • the compound of Formula (I) may be administered in an efficient amount by using a pharmaceutical composition as above-defined.
  • the term “efficient amount” refers to an amount of the compound sufficient to produce the desired therapeutic result.
  • the compound of Formula (I) can be administered in different ways and in different forms.
  • it can be administered in a systematic way, per os, parenterally, by inhalation, or by injection, such as for example intravenously, by intra-muscular route, by subcutaneous route, by transdermal route, by intra-arterial route, etc.
  • Oral administration is the preferential route of administration for pharmaceutical compositions comprising the compound of Formula (I).
  • the frequency and/or dose relative to the administration can be adapted by one of ordinary skill in the art, in function of the patient, the pathology, the form of administration, etc.
  • the compounds of Formula (I) can be administered for the treatment of fibrotic diseases or cancers at doses varying between 0.01 mg and 1 g per administration, preferentially from 1 mg to 100 mg per administration. Administration can be performed daily or even several times per day, if necessary.
  • the invention relates to the use of the compound of Formula (I) for the treatment of a fibrotic disease or cancer, in combination with at least one other therapeutically active agent.
  • the other active agent may in particular be selected from other anti-fibrotic agents or other anti-cancer agents such as Sorafenib.
  • the inventors have shown that the compound of Formula (I) is able to potentiate the pro-apoptotic activity of a pro-apoptotic compound.
  • the invention also relates to the use of the compound of Formula (I) in combination with a pro-apoptotic drug.
  • the compound of Formula (I) may be used in combination with a kinase inhibitor having a pro-apoptotic effect on fibroblasts, such as Staurosporine.
  • the present invention provides methods of treating fibrotic diseases or cancers comprising the administration of the compound of Formula (I), in particular in the form of a pharmaceutical composition containing this compound.
  • hHSC human primary hepatic stellate cells
  • STeCM medium ScienCell cat #5301
  • FBS fetal bovine serum
  • 1% penicillin/streptomycine ScienCell cat #0503
  • stellate cell growth supplement StepCGS; ScienCell cat #5352
  • Culture plastics were coated with Poly-L Lysine (Sigma cat #P4707).
  • hHSC were plated at a density of 1.2 ⁇ 10 4 cells/well into 96-well plates and were cultured overnight at 37° C. and 5% CO 2 , followed by washing of cells with PBS (Invitrogen cat #14190) and replacing the growth medium with a serum-free and SteCGS-free medium for an additional 24 hours.
  • PDGF-induced proliferation assay cells were pre-treated with all compounds for 1 hour before the addition of PDGF-BB (10 ng/mL; R&D Systems cat #520-BB). Treatments were then continued for an additional 20 hours.
  • PDGF-BB 10 ng/mL
  • serum induced proliferation assay cells were pre-treated with compounds for 1 hour before FBS (0.5%) (ScienCell cat #0010) was applied in a SteCGS-free medium for 20 hours.
  • BrdU labeling and detection kit Roche cat #11647229001.
  • BrdU labeling solution was added to cells, followed by incubation for another 4 hours before fixation, addition of nucleases, addition of anti-BrdU-POD and peroxidase substrate.
  • the absorbance at 405 nm was measured using an ELISA plate reader (Tecan).
  • the PDGFR ⁇ phosphorylation in hHSCs was measured using a cell based ELISA kit (R&D Systems cat #KCB1767), according to manufacturer's instructions. Shortly, following the stimulation with PDGF, cells were fixed and permeabilized in the wells. The PDGFR ⁇ phosphorylation was then measured by using a double immunoenzymatic labeling procedure. The cells were simultaneously incubated with two primary antibodies: a phospho-specific antibody that detects the phosphorylation of the PDGFR ⁇ on tyrosine 751 and a control antibody that recognizes both phosphorylated and non-phosphorylated forms of the PDGFR ⁇ .
  • HRP horseradish-peroxidase
  • AP alkaline phosphatase
  • hHSC human primary hepatic stellate cells
  • M-MLV RT Moloney Murine Leukemia Virus Reverse Transcriptase
  • RT buffer 1 ⁇ Invitrogen
  • 1 mM DTT Invitrogen
  • 0.18 mM dNTPs Promega
  • 200 ng pdN6 Amersham
  • 30 U of RNase inhibitor Promega.
  • Quantitative PCR was then carried out using the MyiQ Single-Color Real-Time PCR Detection or the iCycler iQ Multiplex Real-Time PCR Detection System (both systems from Biorad). Briefly, PCR reactions were performed in 96 well plates on 5 ⁇ Lof 5 ⁇ diluted reverse transcription mix using the iQ SYBR Green Supermix kit. The experimental conditions were: 25 ⁇ L of volume reaction, 3 mM of MgCl 2 , and 0.5 ⁇ L each of reverse and forward primers (10 pMol).
  • the induction factor vs TGF ⁇ was determined as follow:
  • Caspase-3 and -7 activities were determined by using the assay from Promega (Promega cat #G8093) following manufacturer's instructions. At the end of the incubation period, 100 ⁇ L of Caspase-Glo® 3/7 Reagent were added to each well containing 100 ⁇ L of blank, negative control cells or treated cells in culture medium. Following cell lysis, the cleavage of the substrate (containing the DEVD sequence) by the activated caspases 3 and 7 was determined by measuring the luminescent signal in a classical plate reader from Tecan. Luminescence was proportional to the amount of caspase activity present in treated cells.
  • reaction cocktails were incubated at 30° C. for 60 minutes.
  • the reaction was stopped with 50 ⁇ l of 2% (v/v) H 3 PO 4 , plates were aspirated and washed two times with 200 ⁇ L 0.9% (w/v) NaCl.
  • Incorporation of 33 P was determined with a microplate scintillation counter (Microbeta, Wallac).
  • the median value of the cpm of three wells with complete reaction cocktails, but without kinase was defined as “low control”. This value reflects unspecific binding of radioactivity to the plate in the absence of protein kinase but in the presence of the substrate. Additionally, for each kinase the median value of the cpm of three other wells with the complete reaction cocktail, but without any compound, was taken as the “high control”, i.e. full activity in the absence of any inhibitor. The difference between high and low control was taken as 100% activity for each kinase.
  • Residual Activity (%) 100 ⁇ [(cpm of compound ⁇ low control)/(high control ⁇ low control)].
  • Compound of formula (I) was given orally at 30 mg/kg/day to the Sprague Dawley rats starting 5 days before the colitis induction by 2,4,6-trinitrobenzenesulfonic acid (TNBS) and until euthanasia.
  • TNBS 2,4,6-trinitrobenzenesulfonic acid
  • TNBS 80 mg/kg in 40% Ethanol
  • Animals were sacrificed 4 days after TNBS administration and the preventive effect of the Compound of formula (I) was assessed using gene expression assay.
  • PCR reactions were performed in 96 well plates on 5 ⁇ L of 5 ⁇ diluted reverse transcription mix using the iQ SYBR Green Supermix kit.
  • the experimental conditions were: 25 ⁇ L of volume reaction, 3 mM of MgCl 2 , and 0.5 ⁇ L each of reverse and forward primers (10 pMol).
  • the induction factor vs TNBS-treated rats was determined as follow:
  • the compound of Formula (I) has anti-proliferative properties and interferes with the functional activation of the PDGFR signaling pathway
  • Ligand-induced receptor homo- or heterodimerization leads to autophosphorylation of specific tyrosine residues within the cytoplasmic domain of PDGFR and to activation of some signal transduction pathways, including phosphatidylinositol 3 kinase (PI3K), Ras-MAPK, Src family kinases and phospholipase C ⁇ (PLC ⁇ ). This results in the stimulation of cell proliferation and survival.
  • PI3K phosphatidylinositol 3 kinase
  • Ras-MAPK Ras-MAPK
  • Src family kinases phospholipase C ⁇
  • FGFR activated pathways resulted in enhanced anti-fibrotic activity in pulmonary fibrosis models as compared to a treatment that inhibits the PDGFR pathway more selectively.
  • the compound of Formula (I) was able to inhibit the kinase activity of selected kinases, including the receptor tyrosine kinases PDGFR, VEGFR, FGFR that are involved in both fibrosis and cancer development.
  • the compound of Formula (I) was able to inhibit, in activated hHSC, the expression of both classical pro-fibrotic genes, such as ⁇ SMA, Col1 ⁇ 1, Col4 ⁇ 1, TGF ⁇ R1, that are induced upon TGF ⁇ 1 treatment and the expression of VEGFR1, VEGFR2, FGFR1, tyrosine kinase receptors that are associated with the development of fibrosis as judged from the previously published sources [63].
  • both classical pro-fibrotic genes such as ⁇ SMA, Col1 ⁇ 1, Col4 ⁇ 1, TGF ⁇ R1, that are induced upon TGF ⁇ 1 treatment and the expression of VEGFR1, VEGFR2, FGFR1, tyrosine kinase receptors that are associated with the development of fibrosis as judged from the previously published sources [63].
  • the gene inhibition profile of the compound of Formula (I) was very similar to that obtained with Sorafenib, a tyrosine kinase inhibitor that targets PDGFR, VEGFR, RAF and KIT. Sorafenib has previously demonstrated important anti-fibrotic activity in preclinical models and clinical efficacy in treatment of such cancers as hepatocellular carcinoma (HCC) and renal cell carcinoma (RCC) [64-66].
  • HCC hepatocellular carcinoma
  • RCC renal cell carcinoma
  • hHSCs primary human hepatic stellate cells
  • TGF ⁇ 1 1 ng/mL
  • Sorafenib Naevaxar
  • the expression of the genes of interest in both treated and untreated hHSCs was determined by the quantitative RT-PCR technique, following 24 hours of exposure to the TGF ⁇ 1.
  • TABLE 4 Compound of Formula (I) inhibits the proliferation of experimental cell lines derived from the selected types of cancer. Inhibition rates are expressed as EC50 values in ⁇ mole/l that were calculated by curve fitting of the experimental data as described in materials and methods.
  • the capacity to induce activated hHSC apoptosis is an important target in fibrotic diseases.
  • the inventors have unexpectedly found that the compound of Formula (I) has modest pro-apoptotic properties, as shown in FIG. 5 , but that it unexpectedly potentiated, in a dose-dependent manner, the pro-apoptotic activity of a broad spectrum kinase inhibitor, Staurosporine.

Landscapes

  • Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Urology & Nephrology (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Dermatology (AREA)
  • Vascular Medicine (AREA)
  • Immunology (AREA)
  • Rheumatology (AREA)
  • Oncology (AREA)
  • Biomedical Technology (AREA)
  • Pulmonology (AREA)
  • Neurology (AREA)
  • Hematology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
US14/760,880 2013-01-18 2014-01-20 Methods of treatment of fibrosis and cancers Abandoned US20150352065A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP13305067.4 2013-01-18
EP13305067 2013-01-18
PCT/EP2014/051060 WO2014111584A1 (en) 2013-01-18 2014-01-20 Methods of treatment of fibrosis and cancers

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2014/051060 A-371-Of-International WO2014111584A1 (en) 2013-01-18 2014-01-20 Methods of treatment of fibrosis and cancers

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US15/586,456 Division US10792277B2 (en) 2013-01-18 2017-05-04 Methods of treatment of fibrosis and cancers

Publications (1)

Publication Number Publication Date
US20150352065A1 true US20150352065A1 (en) 2015-12-10

Family

ID=47628079

Family Applications (2)

Application Number Title Priority Date Filing Date
US14/760,880 Abandoned US20150352065A1 (en) 2013-01-18 2014-01-20 Methods of treatment of fibrosis and cancers
US15/586,456 Active US10792277B2 (en) 2013-01-18 2017-05-04 Methods of treatment of fibrosis and cancers

Family Applications After (1)

Application Number Title Priority Date Filing Date
US15/586,456 Active US10792277B2 (en) 2013-01-18 2017-05-04 Methods of treatment of fibrosis and cancers

Country Status (26)

Country Link
US (2) US20150352065A1 (de)
EP (1) EP2948137B1 (de)
JP (1) JP6218854B2 (de)
KR (1) KR102196277B1 (de)
CN (1) CN105120855B (de)
AU (1) AU2014206781B2 (de)
CA (1) CA2897258C (de)
CY (1) CY1121156T1 (de)
DK (1) DK2948137T3 (de)
EA (1) EA031718B1 (de)
ES (1) ES2702038T3 (de)
HK (1) HK1214773A1 (de)
HU (1) HUE042569T2 (de)
IL (1) IL239715B (de)
LT (1) LT2948137T (de)
MD (1) MD4629B1 (de)
MX (1) MX361565B (de)
NZ (1) NZ709420A (de)
PH (1) PH12015501566B1 (de)
PL (1) PL2948137T3 (de)
PT (1) PT2948137T (de)
SG (1) SG11201505281PA (de)
SI (1) SI2948137T1 (de)
TR (1) TR201819042T4 (de)
WO (1) WO2014111584A1 (de)
ZA (1) ZA201505151B (de)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20190055559A1 (en) * 2016-02-23 2019-02-21 Celgene Corporation Methods of treating intestinal fibrosis using smad7 inhibition
US10792277B2 (en) 2013-01-18 2020-10-06 Genfit Methods of treatment of fibrosis and cancers
US11478440B2 (en) 2017-04-18 2022-10-25 Genfit Combination of Elafibranor or derivatives thereof with an anti-NASH, anti-fibrotic or anti-cholestatic agent

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3830070A1 (de) 2018-08-03 2021-06-09 Genfit Elafibranorsalze
GB201901507D0 (en) * 2019-02-04 2019-03-27 St Georges Hospital Medical School New treatment
AU2021219286A1 (en) 2020-02-10 2022-09-22 Genfit Polymorphs of elafibranor

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011116963A2 (en) * 2010-03-24 2011-09-29 Lipotec S.A. Lipid nanoparticle capsules

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7736677B2 (en) * 2001-06-20 2010-06-15 Metaproteomics, Llc Xanthohumol and tetrahydro-isoalpha acid based protein kinase modulation cancer treatment
FR2841784B1 (fr) * 2002-07-08 2007-03-02 Composition a base de derives de 1,3-diphenylprop-2en-1-one substitues, preparation et utilisations
FR2841900B1 (fr) * 2002-07-08 2007-03-02 Genfit S A Nouveaux derives de 1,3-diphenylprop-2-en-1-one substitues, preparation et utilisations
FR2857361B1 (fr) * 2003-07-08 2005-09-09 Genfit S A PREPARATION DE DERIVES DE 1,3-DIPHENYPROP-2-¼n-1-one
EP2641596B1 (de) * 2009-11-26 2018-05-02 Genfit Verwendung von 1,2-Diphenylprop-2-en-1-on-Derivaten zur Behandlung von Lebererkrankungen
EA031718B1 (ru) 2013-01-18 2019-02-28 Женфит Лечение фиброза и рака, связанных с тирозинкиназой

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011116963A2 (en) * 2010-03-24 2011-09-29 Lipotec S.A. Lipid nanoparticle capsules

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10792277B2 (en) 2013-01-18 2020-10-06 Genfit Methods of treatment of fibrosis and cancers
US20190055559A1 (en) * 2016-02-23 2019-02-21 Celgene Corporation Methods of treating intestinal fibrosis using smad7 inhibition
US11162097B2 (en) * 2016-02-23 2021-11-02 Nogra Pharma Limited Methods of treating intestinal fibrosis using SMAD7 inhibition
US11478440B2 (en) 2017-04-18 2022-10-25 Genfit Combination of Elafibranor or derivatives thereof with an anti-NASH, anti-fibrotic or anti-cholestatic agent

Also Published As

Publication number Publication date
TR201819042T4 (tr) 2019-01-21
KR20150114497A (ko) 2015-10-12
HUE042569T2 (hu) 2019-07-29
US10792277B2 (en) 2020-10-06
US20170239213A1 (en) 2017-08-24
CN105120855B (zh) 2019-05-07
CA2897258C (en) 2021-02-02
ES2702038T3 (es) 2019-02-27
EA031718B1 (ru) 2019-02-28
IL239715B (en) 2019-06-30
EP2948137B1 (de) 2018-11-14
SI2948137T1 (sl) 2019-01-31
LT2948137T (lt) 2018-12-27
CA2897258A1 (en) 2014-07-24
EP2948137A1 (de) 2015-12-02
PL2948137T3 (pl) 2019-02-28
JP2016506913A (ja) 2016-03-07
IL239715A0 (en) 2015-08-31
AU2014206781A1 (en) 2015-07-16
AU2014206781B2 (en) 2018-08-23
CN105120855A (zh) 2015-12-02
HK1214773A1 (zh) 2016-08-05
NZ709420A (en) 2020-04-24
SG11201505281PA (en) 2015-08-28
ZA201505151B (en) 2016-07-27
PT2948137T (pt) 2018-12-20
MD20150076A2 (ro) 2015-11-30
DK2948137T3 (en) 2019-01-07
KR102196277B1 (ko) 2020-12-29
WO2014111584A1 (en) 2014-07-24
JP6218854B2 (ja) 2017-10-25
PH12015501566A1 (en) 2015-09-21
CY1121156T1 (el) 2020-05-29
MX361565B (es) 2018-12-11
EA201591345A1 (ru) 2015-11-30
PH12015501566B1 (en) 2015-09-21
MD4629B1 (ro) 2019-05-31
MX2015009251A (es) 2015-10-15

Similar Documents

Publication Publication Date Title
US10792277B2 (en) Methods of treatment of fibrosis and cancers
US10766881B2 (en) 2-aryl- and 2-heteroaryl-substituted 2-pyridazin-3(2H)-one compounds as inhibitors of FGFR tyrosine kinases
JP7503380B2 (ja) Ａｌｋ阻害剤およびｓｈｐ２阻害剤を含む組合せ医薬
Gaspar et al. Mechanistic evaluation of the novel HSP90 inhibitor NVP-AUY922 in adult and pediatric glioblastoma
ES2408117T3 (es) Compuestos de vinil indazolilo
KR102479693B1 (ko) 조합물
US20160074409A1 (en) Treatment of Skeletal-Related Disorders
TW201316991A (zh) Ｍｅｋ抑制劑與奧諾拉（ａｕｒｏｒａ）ａ激酶選擇性抑制劑之組合
AU2020391220A1 (en) Combination therapy involving diaryl macrocyclic compounds
KR20170134462A (ko) Mdm2 저해제와 btk 저해제의 병용 치료법
CA3213359A1 (en) Alk-5 inhibitors and uses thereof
WO2017156350A1 (en) Methods of cancer treatment
CN101242845A (zh) 使用烷基喹啉类和烷基喹唑啉类协同调制flt3激酶
KR20160090814A (ko) Jak, cdk 및 pim의 억제제를 포함하는 조합 요법
TW202233193A (zh) 包含ｆｌｔ３抑制劑的用於治療白血病的藥物組合物
JP2022507514A (ja) Ｅｒｋインヒビター及びその使用
Harrison et al. The kinase inhibitor O6-cyclohexylmethylguanine (NU2058) potentiates the cytotoxicity of cisplatin by mechanisms that are independent of its effect upon CDK2
Zhang et al. YL143, a novel mutant selective irreversible EGFR inhibitor, overcomes EGFRL858R, T790M‐mutant resistance in vitro and in vivo
Fresnais et al. Safety and Activity of the Combination of Ceritinib and Dasatinib in Osteosarcoma

Legal Events

Date Code Title Description
AS Assignment

Owner name: GENFIT, FRANCE

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:HANF, REMY;HUM, DEAN;WALCZAK, ROBERT;AND OTHERS;SIGNING DATES FROM 20150817 TO 20150824;REEL/FRAME:037302/0259

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION