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
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
  • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/005—Enzyme inhibitors
• • 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
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/50—Pyridazines; Hydrogenated pyridazines
  • A61K31/502—Pyridazines; Hydrogenated pyridazines ortho- or peri-condensed with carbocyclic ring systems, e.g. cinnoline, phthalazine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
  • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• Figure 8 (a) Scatter plots of Compound A Drug Effect (DE) Z-scores from a siRNA screen where the effect of each of 1418 siRNAs on Compound A sensitivity was assessed in BRCA1 defective RPE1 cells. The screen was performed as described above for the CAL51 siRNA screen (b) Amongst the genes whose siRNAs caused increased sensitivity to Compound A (DE ⁇ -2), multiple different siRNAs targeting either FAM35A or REV7 caused sensitivity. By comparison, the DE Z scores for three different control, non targeting, siRNAs in this screen were 1.3 (Allstar control), -0.8 (siCONI) and -0.6 (siCON2). Values shown in the figure are medians from triplicate screens.
• DE Drug Effect
• FIG 11 Graphs demonstrating the effect of the DNA polymerase theta (RoIQ) inhibitor Compound A, the PARP inhibitor olaparib and the control compound staurosporine on the fraction of dead cells in parental (a) and REV7 knockout (KO) (b) SUM149 tumouroids.
• Inhibition of functional activity of RoIQ may be through enzymatic inhibition of its polymerase or helicase domain. In one embodiment, inhibition of RoIQ functional activity is through inhibition of the polymerase domain.
• the RoIQ inhibitor is selected from either of Compounds A or B.
• lymphoid lineage for example acute lymphocytic leukemia [ALL], chronic lymphocytic leukemia [CLL], B-cell lymphomas such as diffuse large B-cell lymphoma [DLBCL], follicular lymphoma, Burkitt’s lymphoma, mantle cell lymphoma, MALT lymphoma, T-cell lymphomas and leukaemias, natural killer [NK] cell lymphomas, Hodgkin’s lymphomas, hairy cell leukaemia, monoclonal gammopathy of uncertain significance, plasmacytoma, multiple myeloma, and post-transplant lymphoproliferative disorders), and haematological malignancies and related conditions of myeloid lineage (for example acute myelogenous leukemia [AML], chronic myelogenous leukemia [CML], chronic
• compositions adapted for parenteral administration include aqueous and non- aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, co-solvents, surface active agents, organic solvent mixtures, cyclodextrin complexation agents, emulsifying agents (for forming and stabilizing emulsion formulations), liposome components for forming liposomes, gellable polymers for forming polymeric gels, lyophilisation protectants and combinations of agents for, inter alia, stabilising the active ingredient in a soluble form and rendering the formulation isotonic with the blood of the intended recipient.
• aqueous and non- aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats, co-solvents, surface active agents, organic solvent mixtures, cyclodextrin complexation agents, emulsifying agents (for forming and stabilizing emulsion formulations), liposome components for forming liposomes, gellable poly
• the compound may be formulated with a carrier and administered in the form of nanoparticles, the increased surface area of the nanoparticles assisting their absorption.
• nanoparticles offer the possibility of direct penetration into the cell.
• Nanoparticle drug delivery systems are described in “Nanoparticle Technology for Drug Delivery”, edited by Ram B Gupta and Uday B. Kompella, Informa Healthcare, ISBN 9781574448573, published 13 th March 2006. Nanoparticles for drug delivery are also described in J. Control. Release, 2003, 91 (1-2), 167-172, and in Sinha et al., Mol. Cancer Ther. August 1, (2006) 5, 1909.
• the pharmaceutical compositions typically comprise from approximately 1% (w/w) to approximately 95% (w/w) active ingredient and from 99% (w/w) to 5% (w/w) of a pharmaceutically acceptable excipient or combination of excipients. Particularly, the compositions comprise from approximately 20% (w/w) to approximately 90%,% (w/w) active ingredient and from 80% (w/w) to 10% of a pharmaceutically acceptable excipient or combination of excipients.
• the pharmaceutical compositions comprise from approximately 1% to approximately 95%, particularly from approximately 20% to approximately 90%, active ingredient.
• Pharmaceutical compositions according to the invention may be, for example, in unit dose form, such as in the form of ampoules, vials, suppositories, pre-filled syringes, dragees, tablets or capsules.
• compositions for topical use and nasal delivery include ointments, creams, sprays, patches, gels, liquid drops and inserts (for example intraocular inserts). Such compositions can be formulated in accordance with known methods.
• formulations for rectal or intra-vaginal administration include pessaries and suppositories which may be, for example, formed from a shaped moldable or waxy material containing the active compound. Solutions of the active compound may also be used for rectal administration.
• a method of treating cancer associated with a Shieldin deficiency which comprises administering a RoIQ inhibitor to a patient in need thereof.
• CBP-501 forkhead translocation inhibitors
• enzastaurin HCI LY317615
• PI3K Inhibitors such as dactolisib (BEZ235), buparlisib (BKM-120; NVP- BKM-120), BYL719, copanlisib (BAY-80-6946), ZSTK-474, CUDC-907, apitolisib (G DC- 0980; RG-7422), pictilisib (pictrelisib, GDC-0941, RG-7321), GDC-0032, GDC-0068, GSK-2636771, idelalisib (formerly CAL-101, GS 1101, GS-1101), MLN1117 (INK1117), MLN0128 (INK128), IPI-145 (INK1197), LY-3023414, ipatasertib, afuresertib, MK-2206, MK
• Monoclonal Antibodies (unconjugated or conjugated to radioisotopes, toxins or other agents), antibody derivatives and related agents, such as anti-CD, anti-VEGFR, anti- HER2, anti-CTLA4, anti-PD-1 or anti-EGFR antibodies, for example rituximab (CD20), ofatumumab (CD20), ibritumomab tiuxetan (CD20), GA101 (CD20), tositumomab (CD20), epratuzumab (CD22), lintuzumab (CD33), gemtuzumab ozogamicin (CD33), alemtuzumab (CD52), galiximab (CD80), trastuzumab (HER2 antibody), pertuzumab (HER2), trastuzumab-DM1 (HER2), ertumaxomab (HER2 and CD3), cetuximab (EGFR), panitum,
• Steroids for example dromostanolone propionate, megestrol acetate, nandrolone (decanoate, phenpropionate), fluoxymestrone or gossypol;
• Cytokine-activating agents include Picibanil, Romurtide, Sizofiran, Virulizin, or Thymosin;
• - agents that prevent or decrease the duration of chemotherapy-associated neutropenia and prevent complications that arise from reduced levels of platelets, red blood cells or white blood cells, for example interleukin-11 (e.g. oprelvekin), erythropoietin (EPO) and analogues thereof (e.g. darbepoetin alfa), colony- stimulating factor analogs such as granulocyte macrophage-colony stimulating factor (GM-CSF) (e.g. sargramostim), and granulocyte-colony stimulating factor (G-CSF) and analogues thereof (e.g. filgrastim, pegfilgrastim),
• interleukin-11 e.g. oprelvekin
• EPO erythropoietin
• analogues thereof e.g. darbepoetin alfa
• colony- stimulating factor analogs such as granulocyte macrophage-colony stimulating factor (GM-CSF) (e
• agents for mucositis e.g. palifermin
• the anti-tumour nucleoside derivative is advantageously administered in a dosage of 200 to 2500 mg per square meter (mg/m 2 ) of body surface area, for example 700 to 1500 mg/m 2 , particularly for 5-FU in a dosage of 200 to 500mg/m 2 , for gemcitabine in a dosage of about 800 to 1200 mg/m 2 and for capecitabine in about 1000 to 2500 mg/m 2 per course of treatment.
• the weight ratio of the compound according to the present invention and the one or more other anticancer agent(s) when given as a combination may be determined by the person skilled in the art. Said ratio and the exact dosage and frequency of administration depends on the particular compound according to the invention and the other anticancer agent(s) used, the particular condition being treated, the severity of the condition being treated, the age, weight, gender, diet, time of administration and general physical condition of the particular patient, the mode of administration as well as other medication the individual may be taking, as is well known to those skilled in the art. Furthermore, it is evident that the effective daily amount may be lowered or increased depending on the response of the treated subject and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. A particular weight ratio for the compound and another anticancer agent may range from 1/10 to 10/1 , more in particular from 1/5 to 5/1, even more in particular from 1/3 to 3/1.
• the compounds of the invention may also be administered in conjunction with non- chemotherapeutic treatments such as radiotherapy, photodynamic therapy, gene therapy; surgery and controlled diets.
• non- chemotherapeutic treatments such as radiotherapy, photodynamic therapy, gene therapy; surgery and controlled diets.
• chemosensitizer is defined as a molecule administered to patients in therapeutically effective amounts to increase the sensitivity of cells to chemotherapy and/or promote the treatment of diseases which are treatable with chemotherapeutics.
• the compound in combination with one or more (e.g. 1 or 2) other therapeutic agents (e.g. anticancer agents) for use in therapy, such as in the prophylaxis or treatment of cancer.
• one or more e.g. 1 or 2
• other therapeutic agents e.g. anticancer agents
• the pharmaceutical composition comprises the compound together with a pharmaceutically acceptable carrier and optionally one or more therapeutic agent(s).
• Shieldin loss represents an effective RoIQ inhibitor patient selection biomarker in an HR-deficient and PARP-resistant setting (see Examples 8 to 12 and Figures 8 to 12).
• Cells were transferred to a cuvette, electroporated using programme EN-113 (HCT 116) or EN-138 (HCC1937) on the 4D nucleofector X unit (Lonza) and recovered into fresh media to a final density of 250,000 cells/mL. 20,000 cells (80pL of suspension) were seeded per well in a white 96-well microplate (Costar 3610) and incubated for 24 hours at 37°C.
• NanoLuciferase levels were detected using the Nano-Glo® Dual-Luciferase® Reporter Assay system (Promega) as per the manufacturer’s instructions, and luminescence was measured with a Clariostar plate reader (BMG Labtech), using the manufacturer’s protocols ‘FireFly’ and ‘NanoLuciferase’. In each well the NanoLuciferase signal was normalised to the Firefly signal, which served as a measure of both cell density and transfection efficiency.
• Membranes were incubated in TBS buffer containing 0.1% Tween 20 (TBST) containing 5% BSA for 2 hours at room temperature, then primary antibody overnight at 4°C. Membranes were washed twice in TBST, then incubated in secondary antibody for 1 hour at room temperature. Membranes were washed four times in TBST, overlaid with ECL detection reagent (GE Healthcare), and exposed in the Amersham AI600 Imager.
• LIG4 Abeam ab193353
• XLF Abeam ab33499
• XRCC4 SCBT sc-271087
• goat-anti-mouse IgG-HRP Thermo 31430
• goat-anti-rabbit IgG-HRP Thermo 31460
• Primary and secondary antibodies were diluted 1:1000 and 1:2000 in 5% BSA, respectively.
• the CRISPR KO screen, sample preparation and data analysis were performed by Horizon Discovery using a CRISPR library against 1965 genes with 10 gRNA’s per gene.
• DLD-1 colon cancer cells were grown in RPMI medium with 10% FBS, infected with the lentiviral library (each viral particle containing Cas9 and sgRNA), selected with puromycin for 2 weeks, and treated with compound B (EC17.1%) or DMSO for 15 days. Synthetic lethality scores were calculated by normalizing the sgRNA count from compound treated cells to DMSO treated control.
• the data was analyzed with QuantStudio Real Time PCR software to calculate the CT (cycle threshold) value for each gene.
• the delta CT was calculated as CT of the test gene minus CT of the housekeeping gene.
• the relative expression was calculated as 2 A (-delta CT) multiplied by 100 to represent the expression of the test gene as a percentage of the expression of the housekeeping gene.
• REV7 KO 22Rv1 cells are significantly more sensitive to RoIQ inhibitor (Compound A, in a and left panels of c) compared to REV7 wild type 22Rv1 parental cells, as evidenced by a decreased relative survival in the REV7 KO cells.
• REV7 KO 22Rv1 cells still retain resistance to a PARP inhibitor (olaparib, in b and right panels of c), as evidenced by a similar surviving fraction in REV7 wild type and REV7 KO cells.
• MDA-MB-436 is a BRCA1 deficient breast cancer cell line.
• the results presented in Figure 13 show that SHLD2 KO MDA-MB-436 cells are significantly more sensitive to RoIQ inhibitor (Compound A, in a and left panels of c) than parental MDA-MB-436 cells, as evidenced by a decreased relative survival.
• a trend for increased resistance of SHLD2 KO MDA-MB-436 to a PARP inhibitor (olaparib, in b and right panels of c) is also observed, as evidenced by an increased relative survival.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Chemical & Material Sciences (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Medicinal Chemistry (AREA)
• Pharmacology & Pharmacy (AREA)
• General Health & Medical Sciences (AREA)
• Epidemiology (AREA)
• Immunology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Engineering & Computer Science (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Organic Chemistry (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Priority Applications (7)

Applications Claiming Priority (1)

Publications (1)

Family

ID=74570294

Family Applications (1)

Country Status (6)

Cited By (5)

Citations (2)

Family Cites Families (1)

• 2019
  • 2019-08-09 CA CA3149112A patent/CA3149112A1/en active Pending
  • 2019-08-09 JP JP2022507857A patent/JP2023501038A/ja not_active Ceased
  • 2019-08-09 CN CN201980101223.8A patent/CN114728036A/zh active Pending
  • 2019-08-09 WO PCT/GB2019/052243 patent/WO2021028644A1/en not_active Ceased
  • 2019-08-09 US US17/597,940 patent/US20220387544A1/en not_active Abandoned
  • 2019-08-09 EP EP19755666.5A patent/EP4010002A1/en not_active Withdrawn
• 2024
  • 2024-01-31 JP JP2024012755A patent/JP2024059632A/ja active Pending

Patent Citations (2)

Non-Patent Citations (30)

Also Published As

Similar Documents

Legal Events