WO2014013231A1 - Matériaux et procédés pour le traitement d'un cancer à mutation ou déficience en pten - Google Patents

Matériaux et procédés pour le traitement d'un cancer à mutation ou déficience en pten Download PDF

Info

Publication number
WO2014013231A1
WO2014013231A1 PCT/GB2013/051878 GB2013051878W WO2014013231A1 WO 2014013231 A1 WO2014013231 A1 WO 2014013231A1 GB 2013051878 W GB2013051878 W GB 2013051878W WO 2014013231 A1 WO2014013231 A1 WO 2014013231A1
Authority
WO
WIPO (PCT)
Prior art keywords
pten
inhibitor
nlk
cancer
deficient
Prior art date
Application number
PCT/GB2013/051878
Other languages
English (en)
Inventor
Alan Ashworth
Christopher James LORD
Ana Maria MENDES PEREIRA
Original Assignee
The Institute Of Cancer Research: Royal Cancer Hospital
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 The Institute Of Cancer Research: Royal Cancer Hospital filed Critical The Institute Of Cancer Research: Royal Cancer Hospital
Publication of WO2014013231A1 publication Critical patent/WO2014013231A1/fr

Links

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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03016—Phosphoprotein phosphatase (3.1.3.16), i.e. calcineurin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03048—Protein-tyrosine-phosphatase (3.1.3.48)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03067—Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase (3.1.3.67)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.

Definitions

  • the present invention relates to materials and methods for treating PTEN mutated or deficient cancer using Nemo-Like kinase (NLK) inhibitors, and to methods of screening for agents for treating PTEN mutated or deficient cancer.
  • NLK Nemo-Like kinase
  • SSL can occur between genes acting in the same biochemical pathway or in distinct but compensatory pathways .
  • Tankyrase 1 inhibitors may be used for treating BRCA-associated cancers (WO2009/027650 ) and that DNA polymerase POLp inhibitors may be used for treating DNA mismatch repair (MMR) deficient cancers (WO2009/027641 ) .
  • MMR DNA mismatch repair
  • SL synthetic lethality
  • RNAi RNA interference
  • Phosphatase and tensin homolog (“PTEN”) is a gene that has been identified as a tumor suppressor through the action of its phosphatase product and is mutated in a large number of cancers at high frequency.
  • the protein encoded this gene is a
  • phosphatases Unlike most of the protein tyrosine phosphatases, this protein preferentially dephosphorylates phosphoinositide substrates . It negatively regulates intracellular levels of phosphatidylinositol-3 , 4 , 5-trisphosphate in cells and functions as a tumor suppressor by negatively regulating AKT/PKB and mTOR signalling pathways.
  • the HUGO Gene Symbol report for PTEN can be found at
  • Nemo-Like kinase (NLK) , inhibitors of which can be used in accordance with the present invention for the treatment of PTEN mutated or deficient cancer according to the present invention, is a 527 amino acid.
  • amino acid sequence of human FOXOl is set out in SEQ ID NO: 2.
  • the present invention provides an inhibitor of a Nemo-Like kinase (NLK) for use in a method of treating an individual having cancer, wherein the cancer is a Phosphatase and Tensin Homolog (PTEN) mutated or deficient cancer .
  • NLK Nemo-Like kinase
  • PTEN Tensin Homolog
  • the present invention provides the use of a Nemo-Like kinase (NLK) inhibitor in the preparation of a NLK inhibitor.
  • NLK Nemo-Like kinase
  • the present invention provides a method of treating an individual having a PTEN mutated or deficient cancer, the method comprising administering to the individual a
  • NLK Nemo-Like kinase
  • the present invention provides a method of screening for agents useful in the treatment of a PTEN mutated or deficient cancer, the method employing first and second cell lines, wherein the first cell line is PTEN and the second cell line is PTEN proficient, the method comprising:
  • the present invention provides a method of screening for agents useful in the treatment of PTEN mutated or deficient cancer, the method comprising:
  • NLK Nemo-Like kinase
  • the present invention provides a method which comprises having identified a candidate agent useful for the treatment of a PTEN mutated or deficient cancer according to a method as described herein, the further step of manufacturing the compound in bulk and/or formulating the agent in a pharmaceutical composition .
  • NPI values from HCT116 P TEN ⁇ / ⁇ and HCT116 PTEN + + screens. Blue dots corresponded to siPLK effects and red dots corresponded to siCON negative control effects. NPI values below the trend line shown were considered as candidate synthetic lethalities .
  • NLK C-20: sc-8210, Santa Cruz Biotechnology
  • PLK4 #3258, Cell Signaling
  • TTK C-19: sc-540, Santa Cruz Biotechnology
  • ⁇ -Actin 119: sc-1616, Santa Cruz Biotechnology
  • transfection cells were fixed in PBS containing 2% (w/v) formaldehyde and 0.2% (v/v) glutaraldehyde , rinsed and incubated overnight at 37°C in a solution containing X-gal according to manufacturer instructions (Senescence ⁇ -Galactosidase Staining Kit, Cell Signaling Technology) . After aspirating staining solution, cells were overlaid with 70% (v/v) glycerol and stored at 4°C until reading absorbance at 595 nm (A 595 ) in a Victor X5 plate reader (Perkin Elmer) . A 595 values were normalised to the A 595 signal in PTEN proficient cells transfected with siCONpool#2 transfected cells to give relative senescence levels in each experimental sample as shown. siRNA targeting SKP2 was used a positive control (17),
  • Derivatives may be designed, based on a lead compound, by modifying one or more substituents or functional groups compared to the lead compound, for example by replacing these with alternative substituents or groups which are expected to have the same or improved physiological effect.
  • the use of derivatives having such modifications is well known to those in the art.
  • NLK Nemo-Like kinase
  • a candidate inhibitor for example, may be a "functional analogue" of a peptide fragment or other compound which inhibits the component.
  • a functional analogue has the same functional activity as the peptide or other compound in question. Examples of such analogues include chemical compounds which are modelled to resemble the three dimensional structure of the component in an area which contacts another component, and in particular the arrangement of the key amino acid residues as they appear .
  • Another class of inhibitors useful for treatment of PTEN mutated or deficient cancer includes nucleic acid inhibitors of a Nemo- Like kinase (NLK) , or the complements thereof, which inhibit activity or function by down-regulating production of active polypeptide. This can be monitored using conventional methods well known in the art, for example by screening using real time PCR as described in the examples.
  • NLK Nemo- Like kinase
  • Anti-sense oligonucleotides may be designed to hybridise to the complementary sequence of nucleic acid, pre-mRNA or mature mRNA, interfering with the production of the base excision repair pathway component so that its expression is reduced or completely or substantially completely prevented.
  • anti-sense techniques may be used to target control sequences of a gene, e.g. in the 5' flanking sequence, whereby the anti-sense oligonucleotides can interfere with expression control sequences.
  • the construction of anti-sense sequences and their use is described for example in Peyman & Ulman, Chemical Reviews, 90:543-584, 1990 and Crooke, Ann. Rev. Pharmacol. Toxicol., 32:329-376, 1992.
  • Oligonucleotides may be generated in vitro or ex vivo for administration or anti-sense RNA may be generated in vivo within cells in which down-regulation is desired.
  • double-stranded DNA may be placed under the control of a promoter in a "reverse orientation" such that transcription of the anti-sense strand of the DNA yields RNA which is complementary to normal mRNA
  • the complete sequence corresponding to the coding sequence in reverse orientation need not be used.
  • fragments of sufficient length may be used. It is a routine matter for the person skilled in the art to screen fragments of various sizes and from various parts of the coding or flanking sequences of a gene to optimise the level of anti-sense inhibition. It may be advantageous to include the initiating methionine ATG codon, and perhaps one or more nucleotides upstream of the initiating codon.
  • a suitable fragment may have about 14-23 nucleotides, e.g., about 15, 16 or 17 nucleotides.
  • RNAi RNA interference
  • RNA interference is a two-step process. First, dsRNA is cleaved within the cell to yield short interfering RNAs (siRNAs) of about 21-23nt length with 5' terminal phosphate and 3' short overhangs ( ⁇ 2nt) . The siRNAs target the corresponding mRNA sequence specifically for destruction (Zamore, Nature Structural Biology, 8, 9, 746-750, 2001.
  • siRNA duplexes have been shown to specifically suppress expression of endogenous and heterologeous genes in a wide range of mammalian cell lines
  • nucleic acid is used which on transcription produces a ribozyme, able to cut nucleic acid at a specific site and therefore also useful in influencing gene expression, e.g., see Kashani-Sabet & Scanlon, Cancer Gene Therapy, 2(3): 213-223, 1995 and Mercola & Cohen, Cancer Gene Therapy, 2(1): 47-59, 1995.
  • Double- stranded RNA (dsRNA) -dependent post transcriptional silencing also known as RNA interference (RNAi)
  • RNAi Double- stranded RNA
  • RNAi RNA interference
  • a 20-nt siRNA is generally long enough to induce gene-specific silencing, but short enough to evade host response. The decrease in expression of targeted gene products can be extensive with 90% silencing induced by a few molecules of siRNA.
  • RNA sequences are termed “short or small interfering RNAs” (siRNAs) or “microRNAs” (miRNAs) depending on their origin. Both types of sequence may be used to down- regulate gene expression by binding to complimentary RNAs and either triggering mRNA elimination (RNAi) or arresting mRNA translation into protein.
  • siRNA are derived by processing of long double stranded RNAs and when found in nature are typically of exogenous origin.
  • miRNA are examples of interfering RNAs
  • siRNA and miRNA can inhibit the
  • siRNA ligands are typically double stranded and, in order to optimise the effectiveness of RNA mediated down-regulation of the function of a target gene, it is preferred that the length of the siRNA molecule is chosen to ensure correct recognition of the siRNA by the RISC complex that mediates the recognition by the siRNA of the mRNA target and so that the siRNA is short enough to reduce a host response.
  • miRNA ligands are typically single stranded and have regions that are partially complementary enabling the ligands to form a hairpin.
  • miRNAs are RNA genes which are transcribed from DNA, but are not translated into protein. A DNA sequence that codes for a miRNA gene is longer than the miRNA. This DNA sequence includes the miRNA sequence and an approximate reverse
  • RNA ligands intended to mimic the effects of siRNA or miRNA have between 10 and 40 ribonucleotides (or synthetic analogues thereof) , more preferably between 17 and 30
  • ribonucleotides more preferably between 19 and 25
  • ribonucleotides and most preferably between 21 and 23
  • the molecule may have symmetric 3 ' overhangs, e.g. of one or two (ribo) nucleotides, typically a UU of dTdT 3' overhang.
  • siRNA and miRNA sequences can be synthetically produced and added exogenously to cause gene downregulation or produced using expression systems (e.g. vectors). In a preferred embodiment the siRNA is synthesized synthetically.
  • Longer double stranded RNAs may be processed in the cell to produce siRNAs (e.g. see Myers, Nature Biotechnology, 21: 324- 328, 2003) .
  • the longer dsRNA molecule may have symmetric 3' or 5' overhangs, e.g. of one or two ( ribo ) nucleotides , or may have blunt ends.
  • the longer dsRNA molecules may be 25 nucleotides or longer.
  • the longer dsRNA molecules are between 25 and 30 nucleotides long. More preferably, the longer dsRNA molecules are between 25 and 27 nucleotides long.
  • the longer dsRNA molecules are 27 nucleotides in length.
  • dsRNAs 30 nucleotides or more in length may be expressed using the vector pDECAP (Shinagawa et al . , Genes and Dev., 17: 1340-5, 2003) .
  • shRNA short hairpin RNA molecule
  • shRNAs are more stable than synthetic siRNAs .
  • a shRNA consists of short inverted repeats separated by a small loop sequence. One inverted repeat is complimentary to the gene target.
  • the shRNA is processed by DICER into a siRNA which degrades the target gene mRNA and suppresses expression.
  • the shRNA is produced endogenously (within a cell) by transcription from a vector.
  • shRNAs may be produced within a cell by
  • RNA polymerase III promoter such as the human HI or 7SK promoter or a RNA polymerase II promoter.
  • the shRNA may be synthesised exogenously (in vitro) by transcription from a vector.
  • the shRNA may then be introduced directly into the cell.
  • the shRNA sequence is between 40 and 100 bases in length, more preferably between 40 and 70 bases in length.
  • the stem of the hairpin is preferably between 19 and 30 base pairs in length.
  • the stem may contain G-U pairings to stabilise the hairpin structure.
  • the siRNA, longer dsRNA or miRNA is produced endogenously (within a cell) by transcription from a vector.
  • the vector may be introduced into the cell in any of the ways known in the art.
  • expression of the RNA sequence can be regulated using a tissue specific promoter.
  • the siRNA, longer dsRNA or miRNA is produced
  • siRNA molecules may be synthesized using standard solid or solution phase synthesis techniques, which are known in the art.
  • Linkages between nucleotides may be phosphodiester bonds or alternatives, e.g., linking groups of the formula P(0)S, (thioate); P(S)S, (dithioate ) ; P(0)NR'2; P(0)R'; P(0)OR6; CO; or CONR'2 wherein R is H (or a salt) or alkyl (1-12C) and R6 is alkyl (1-9C) is joined to adjacent nucleotides through-O-or-S- .
  • siRNA molecules containing them may confer advantageous properties on siRNA molecules containing them.
  • modified bases may increase the stability of the siRNA molecule, thereby reducing the amount required for
  • modified bases may also provide siRNA molecules, which are more, or less, stable than unmodified siRNA.
  • Modified nucleotides include alkylated purines and pyrimidines, acylated purines and pyrimidines, and other heterocycles . These classes of pyrimidines and purines are known in the art and include pseudoisocytosine , N4,N4- ethanocytosine , 8-hydroxy-N6-methyladenine , 4-acetylcytosine, 5- ( carboxyhydroxylmethyl ) uracil, 5 fluorouracil , 5-bromouracil, 5- carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyl uracil, dihydrouracil, inosine, N6-isopentyl-adenine , 1- methyladenine, 1-methylpseudouracil, 1-methylguanine , 2,2- dimethylguanine, 2methyladenine, 2-methylguanine, 3- methylcytosine , 5-methylcytosine , N6-
  • Common tumor associated PTEN mutations include C.3880G,
  • the present invention is particularly applicable to the treatment of cancer cells that are also FOXOl proficient, e.g. in which the activity of FOXOl is preferably at least 50% normal wild-type activity, more preferably at least 75% wild-type activity and most preferably at least 90% wild-type activity .
  • the activity of the PTEN may be determined by using techniques well known in the art such as Western blot analysis,
  • the determination of PTEN gene expression may involve determining the presence or amount of PTEN mRNA in a sample. Methods for doing this are well known to the skilled person. By way of example, they include determining the presence of PTEN mRNA (i) using a labelled probe that is capable of hybridising to the PTEN nucleic acid; and/or (ii) using PCR involving one or more primers based on a PTEN nucleic acid sequence to determine whether the PTEN transcript is present in a sample.
  • the probe may also be immobilised as a sequence included in a microarray.
  • detecting PTEN mRNA is carried out by extracting RNA from a sample of the tumour and measuring PTEN expression specifically using quantitative real time RT-PCR.
  • the expression of PTEN could be assessed using RNA extracted from a tumour sample using microarray analysis, which measures the levels of mRNA for a group of genes using a plurality of probes immobilised on a substrate to form the array. The determination of whether the cells are express FOXOl and hence are FOXOl proficient may be done in an analogous manner.
  • deficient cancer can be carried out by analysis of PTEN protein expression, for example to examining whether reduced levels of PTEN protein are expressed or whether the PTEN protein contains one or more mutations.
  • the determination of whether the cells are FOXOl proficient may be done in an analogous manner.
  • the method for determining the presence of PTEN protein and/or FOXOl protein may be carried out on tumour samples, for example using immunohistochemical (IHC) analysis.
  • IHC analysis can be carried out using paraffin fixed samples or frozen tissue samples, and generally involves staining the samples to highlight the presence and location of PTEN protein and/or FOXOl protein.
  • compositions that additionally comprise with one or more pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, stabilisers, preservatives,
  • derivatives of the therapeutic agents includes salts, coordination complexes, esters such as in vivo
  • hydrolysable esters free acids or bases, hydrates, prodrugs or lipids, coupling partners.
  • Salts of the compounds of the invention are preferably
  • Esters can be formed between hydroxyl or carboxylic acid groups present in the compound and an appropriate carboxylic acid or alcohol reaction partner, using techniques well known in the art.
  • Coupled derivatives include coupling partners of the compounds in which the compounds is linked to a coupling partner, e.g. by being chemically coupled to the compound or physically
  • coupling partners include a label or reporter molecule, a supporting substrate, a carrier or transport molecule, an effector, a drug, an antibody or an inhibitor.
  • Coupling partners can be covalently linked to compounds of the invention via an appropriate functional group on the compound such as a hydroxyl group, a carboxyl group or an amino group.
  • Other derivatives include formulating the compounds with liposomes .
  • pharmaceutically acceptable includes compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgement, suitable for use in contact with the tissues of a subject (e.g. human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable
  • the active agents disclosed herein for the treatment of PTEN mutated or deficient cancer according to the present invention are preferably for administration to an individual in a
  • composition may be
  • formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing the active compound into association with a carrier, which may constitute one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into
  • agents disclosed herein for the treatment of PTEN mutated or deficient cancer may be administered to a subject by any combination
  • pulmonary e.g. by inhalation or insufflation therapy using, e.g. an aerosol, e.g. through mouth or nose); rectal; vaginal;
  • parenteral for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial,
  • intracardiac intrathecal
  • intraspinal intracapsular
  • Formulations suitable for oral administration e.g., by
  • ingestion may be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in- water liquid emulsion or a water-in-oil liquid emulsion; as a bolus; as an electuary; or as a paste.
  • Formulations suitable for parenteral administration include aqueous and non-aqueous isotonic, pyrogen-free , sterile injection solutions which may contain anti-oxidants , buffers, preservatives, stabilisers, bacteriostats , and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and nonaqueous sterile suspensions which may include suspending agents and thickening agents, and liposomes or other microparticulate systems which are designed to target the compound to blood components or one or more organs .
  • Suitable isotonic vehicles for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection.
  • concentration of the active compound in the solution is from about 1 ng/ml to about 10 ⁇ g/ml, for example from about 10 ng/ml to about 1 ⁇ g/ml.
  • the formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.
  • Formulations may be in the form of liposomes or other microparticulate systems which are designed to target the active compound to blood components or one or more organs .
  • compositions comprising agents disclosed herein for the treatment PTEN mutated or deficient cancer may be used in the methods described herein in combination with standard chemotherapeutic regimes or in conjunction with radiotherapy.
  • chemotherapeutic agents include Amsacrine (Amsidine) , Bleomycin, Busulfan, Capecitabine (Xeloda) , Carboplatin, Carmustine (BCNU) , Chlorambucil (Leukeran) , Cisplatin, Cladribine (Leustat) ,
  • Clofarabine (Evoltra) , Crisantaspase (Erwinase),
  • Cyclophosphamide Cytarabine (ARA-C)
  • DTIC dacarbazine
  • Dactinomycin Actinomycin D
  • Daunorubicin Daunorubicin, Docetaxel (Taxotere)
  • Doxorubicin Epirubicin
  • Etoposide Vepesid, VP-16
  • Fludarabine Fludara
  • Fluorouracil 5-FU
  • Gemcitabine Gemzar
  • Hydroxyurea Hydrourea
  • Idarubicin Zavedos
  • tumourigenesis at least in part, by causing downstream
  • tumour-specific loss-of-function mutations raises the possibility of identifying PTEN synthetic lethal interactions that could be exploited therapeutically.
  • RNA interference represents a straightforward method to identifying SLs in a relatively unbiased fashion (4).
  • HTS high-throughput screening
  • NLK Nemo-Like Kinase
  • PLK4 Polyo-Like Kinase 4
  • TTK Monopolar Spindle 1
  • NLK and PLK4 SLs were examined in a wide panel of genetically heterogenous PTEN deficient or proficient tumour cell lines (Table 1) .
  • Table 1 we used a combination of two validated siRNAs to target each gene, alongside appropriate positive (siPLKl) and negative (siCON) controls ( Figure S2) .
  • Silencing of either NLK or PLK had a greater cell inhibitory effect in the PTEN deficient cohort when compared to the PTEN proficient group ( Figure 2C, 2D), an effect especially evident for NLK silencing.
  • FOXOl Forkhead- box-01' isoform
  • FOXOl can exert tumour suppressive effects by mediating the transcription of genes such as p21, p27, and BIM that promote quiescence, senescence or cell death.
  • this tumour suppressive function of FOXOl can in turn be circumvented by the AKT
  • NLK inhibition could be synthetically lethal with PTEN deficiency as it causes reactivation of FOXOl and therefore cellular senescence.
  • PTEN/NLK synthetic lethality was FOXOl dependent, we transfected PTEN null cells with both NLK and FOXOl siRNA and assessed the effect on cell inhibition.
  • FOXOl siRNA Whilst FOXOl siRNA alone did not selectively target PTEN null cells and NLKl siRNA caused PTEN synthetic lethality (as before) , FOXOl siRNA reversed the PTEN/NLK synthetic lethality ( Figure 3A, Figure S3), supporting the hypothesis that the PTEN/NLK SL is FOXOl dependent. NLK silencing also caused an increase in nuclear FOXOl localisation ( Figure 3B) , again supporting the hypothesis. Furthermore, NLK silencing induced senescence in PTEN deficient cells when compared to PTEN proficient
  • NLK silencing inhibits PTEN deficient tumour cells.
  • Our mechanistic dissection of this effect suggests that re-instatement of a FOXOl-driven process, possibly cellular senescence that normally suppresses oncogenesis, could explain the PTEN selective effect of NLK inhibition.
  • others have already identified compounds that cause the translocation FOXOl to the nucleus (16) .
  • the data presented here suggests that inhibition of NLK could provide a similar target for drug development.
  • PTEN deficiency could be used as a biomarker to identify patients likely to respond to a clinical NLK inhibitor.
  • VHPVQQHTSS AAAAAAAAAA AAAMLNPGQQ QPYFPSPAPG QAPGPAAAAP AQVQAAAAAT

Abstract

L'invention concerne des matériaux et des procédés pour le traitement d'un cancer à mutation ou déficience en PTEN en utilisant des inhibiteurs de NLK (Nemo-Like kinase), et des procédés de criblage d'agents pour le traitement d'un cancer à mutation ou déficience en PTEN. Le cancer à mutation ou déficience en PTEN présente éventuellement également une abondance de FOXO1.
PCT/GB2013/051878 2012-07-16 2013-07-15 Matériaux et procédés pour le traitement d'un cancer à mutation ou déficience en pten WO2014013231A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201261671845P 2012-07-16 2012-07-16
GB1212690.0 2012-07-16
GBGB1212690.0A GB201212690D0 (en) 2012-07-16 2012-07-16 Materials and methods for treating pten mutated or deficient cancer
US61/671,845 2012-07-16

Publications (1)

Publication Number Publication Date
WO2014013231A1 true WO2014013231A1 (fr) 2014-01-23

Family

ID=46799737

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2013/051878 WO2014013231A1 (fr) 2012-07-16 2013-07-15 Matériaux et procédés pour le traitement d'un cancer à mutation ou déficience en pten

Country Status (2)

Country Link
GB (1) GB201212690D0 (fr)
WO (1) WO2014013231A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022005879A1 (fr) * 2020-07-01 2022-01-06 The Board Of Trustees Of The Leland Stanford Junior University Procédés de traitement de syndromes de défaillance de la moelle osseuse et compositions à utiliser dans ceux-ci

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007044571A2 (fr) * 2005-10-06 2007-04-19 President And Fellows Of Harvard College Procedes permettant d'identifier des proteines essentielles a la proliferation des cellules humaines, et agents therapeutiques diriges contre ces proteines
US20070265333A1 (en) * 2006-05-11 2007-11-15 Hong Fu Macrocyclic kinase inhibitors
US20100048552A1 (en) * 2006-05-11 2010-02-25 Irm Llc Compounds and compositions as protein kinase inhbitors
WO2011106168A1 (fr) * 2010-02-24 2011-09-01 Dcam Pharma Inc Composés de purine pour le traitement de maladies auto-immunes et démyélinisantes
WO2011118994A2 (fr) * 2010-03-25 2011-09-29 가톨릭대학교 산학협력단 Gène nlk en tant que marqueur pour le diagnostic du cancer du foie et en tant qu'agent thérapeutique correspondant
WO2012000103A1 (fr) * 2010-07-02 2012-01-05 University Health Network Méthodes de ciblage de maladies a mutation de pten et compositions prévues à cet effet
WO2012168721A1 (fr) * 2011-06-10 2012-12-13 The Institute Of Cancer Research: Royal Cancer Hospital Substances et procédés pour traiter un cancer à déficience ou à mutation du gène pten

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007044571A2 (fr) * 2005-10-06 2007-04-19 President And Fellows Of Harvard College Procedes permettant d'identifier des proteines essentielles a la proliferation des cellules humaines, et agents therapeutiques diriges contre ces proteines
US20070265333A1 (en) * 2006-05-11 2007-11-15 Hong Fu Macrocyclic kinase inhibitors
US20100048552A1 (en) * 2006-05-11 2010-02-25 Irm Llc Compounds and compositions as protein kinase inhbitors
WO2011106168A1 (fr) * 2010-02-24 2011-09-01 Dcam Pharma Inc Composés de purine pour le traitement de maladies auto-immunes et démyélinisantes
WO2011118994A2 (fr) * 2010-03-25 2011-09-29 가톨릭대학교 산학협력단 Gène nlk en tant que marqueur pour le diagnostic du cancer du foie et en tant qu'agent thérapeutique correspondant
WO2012000103A1 (fr) * 2010-07-02 2012-01-05 University Health Network Méthodes de ciblage de maladies a mutation de pten et compositions prévues à cet effet
WO2012168721A1 (fr) * 2011-06-10 2012-12-13 The Institute Of Cancer Research: Royal Cancer Hospital Substances et procédés pour traiter un cancer à déficience ou à mutation du gène pten

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ANA M. MENDES-PEREIRA ET AL: "NLK Is a Novel Therapeutic Target for PTEN Deficient Tumour Cells", PLOS ONE, vol. 7, no. 10, 29 October 2012 (2012-10-29), pages e47249, XP055086547, DOI: 10.1371/journal.pone.0047249 *
DATABASE WPI Week 201166, Derwent World Patents Index; AN 2011-M37929, XP002716963 *
JUNG KWANG HWA ET AL: "Targeted Disruption of Nemo-Like Kinase Inhibits Tumor Cell Growth by Simultaneous Suppression of Cyclin D1 and CDK2 in Human Hepatocellular Carcinoma", JOURNAL OF CELLULAR BIOCHEMISTRY, vol. 110, no. 3, June 2010 (2010-06-01), pages 687 - 696, XP002716964, ISSN: 0730-2312 *
MIN SUP SONG ET AL: "Nuclear PTEN Regulates the APC-CDH1 Tumor-Suppressive Complex ina Phosphatase-Independent Manner", CELL, CELL PRESS, US, vol. 144, no. 2, 15 December 2010 (2010-12-15), pages 187 - 199, XP028152918, ISSN: 0092-8674, [retrieved on 20101217], DOI: 10.1016/J.CELL.2010.12.020 *
ZHUJUN TAN ET AL: "NLK is a key regulator of proliferation and migration in gallbladder carcinoma cells", MOLECULAR AND CELLULAR BIOCHEMISTRY, KLUWER ACADEMIC PUBLISHERS, BO, vol. 369, no. 1 - 2, 24 June 2012 (2012-06-24), pages 27 - 33, XP035104166, ISSN: 1573-4919, DOI: 10.1007/S11010-012-1365-0 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022005879A1 (fr) * 2020-07-01 2022-01-06 The Board Of Trustees Of The Leland Stanford Junior University Procédés de traitement de syndromes de défaillance de la moelle osseuse et compositions à utiliser dans ceux-ci

Also Published As

Publication number Publication date
GB201212690D0 (en) 2012-08-29

Similar Documents

Publication Publication Date Title
Kok et al. Cancer-derived exosomes: their role in cancer biology and biomarker development
EP2718436B1 (fr) Substances et procédés pour traiter un cancer à déficience ou à mutation du gène pten
JP6599334B2 (ja) 血中循環腫瘍細胞に関する方法およびアッセイ
Wang et al. miR-363-3p inhibits tumor growth by targeting PCNA in lung adenocarcinoma
Lamers et al. Knockdown of survivin (BIRC5) causes apoptosis in neuroblastoma via mitotic catastrophe
Yoshida et al. Clinical and functional significance of intracellular and extracellular microRNA-25-3p in osteosarcoma
US20130059903A1 (en) Compositions and Methods for Characterizing Breast Cancer
US9322016B2 (en) MicroRNA-based methods and assays for osteosarcoma
MX2015005441A (es) Letalidad sintetica y el tratamiento de cancer.
JP6543612B2 (ja) 大腸癌の治療剤、及び大腸癌患者の予後の予測方法
Wu et al. XIAP 3′-untranslated region as a ceRNA promotes FSCN1 function in inducing the progression of breast cancer by binding endogenous miR-29a-5p
Blumental-Perry et al. Retrograde signaling by a mtDNA-encoded non-coding RNA preserves mitochondrial bioenergetics
Lei et al. circTLK1 facilitates the proliferation and metastasis of renal cell carcinoma by regulating miR-495-3p/CBL axis
CA2816929C (fr) Letalite synthetique dans le cancer
WO2018228990A1 (fr) Inhibiteurs de kinases dépendantes des cyclines 4/6 destinés à être utilisés dans des méthodes de traitement du cancer
WO2009027650A1 (fr) Matières et procédés pour exploiter la létalité synthétique dans des cancers associés à brca
Luo et al. LncRNA CASC2 inhibits proliferation and migration of adenocarcinoma cells via miR‐4735‐3p and mTOR
WO2014013231A1 (fr) Matériaux et procédés pour le traitement d'un cancer à mutation ou déficience en pten
Liu et al. The deubiquitinating enzyme MINDY2 promotes pancreatic cancer proliferation and metastasis by stabilizing ACTN4 expression and activating the PI3K/AKT/mTOR signaling pathway
WO2018224536A1 (fr) Inhibiteurs de parp destinés à être utilisés pour le traitement du cancer
Zhou et al. Knockdown of the long non‑coding RNA CACNA1G‑AS1 enhances cytotoxicity and apoptosis of human diffuse large B cell lymphoma by regulating miR‑3160‑5p
WO2012168720A1 (fr) Substances et procédés pour traiter un cancer positif aux récepteurs des oestrogènes α (er)
US10905708B2 (en) MicroRNA-based methods and assays for osteocarcinoma
US20220218708A1 (en) Methods for treating smarcb1 deficient cancer or pazopanib resistant cancer
JP2010501162A (ja) 肺癌の治療標的及び予後指標としてのｉｍｐ−１癌遺伝子

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13740339

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 13740339

Country of ref document: EP

Kind code of ref document: A1