[go: up one dir, main page]

WO2013177349A2 - Quinazolinediones as tankyrase inhibitors - Google Patents

Quinazolinediones as tankyrase inhibitors Download PDF

Info

Publication number
WO2013177349A2
WO2013177349A2 PCT/US2013/042332 US2013042332W WO2013177349A2 WO 2013177349 A2 WO2013177349 A2 WO 2013177349A2 US 2013042332 W US2013042332 W US 2013042332W WO 2013177349 A2 WO2013177349 A2 WO 2013177349A2
Authority
WO
WIPO (PCT)
Prior art keywords
methyl
mmol
dioxo
cyclohexane
mixture
Prior art date
Application number
PCT/US2013/042332
Other languages
French (fr)
Other versions
WO2013177349A3 (en
Inventor
Richard M. Keenan
Aaron Bayne Miller
Donghui Qin
Original Assignee
Glaxosmithkline Llc
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 Glaxosmithkline Llc filed Critical Glaxosmithkline Llc
Publication of WO2013177349A2 publication Critical patent/WO2013177349A2/en
Publication of WO2013177349A3 publication Critical patent/WO2013177349A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/95—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in positions 2 and 4
    • C07D239/96—Two oxygen atoms

Definitions

  • This invention relates to novel quinazolinediones which are inhibitors of tankyrases (TNKS1 and TNKS2), to pharmaceutical compositions containing them, and to their use in therapy for the treatment of cancers, fibrosis and other hyperproliferative diseases through this mechanism.
  • TNKS1 and TNKS2 tankyrases
  • the tankyrases (TNKS1 and TNKS2) belong to the poly ADP-ribose polymerase (PARP) family of enzymes and act via mono- or poly-ADP-ribosylation (parsylation) of substrate proteins.
  • PARP poly ADP-ribose polymerase
  • the tankyrases also contain the ANK domain, which contains 16-24 ankyrin repeats.
  • the ANK domain interacts with a variety of proteins, including the telomeric protein Telomere Repeat binding Factor- 1 (TRF-1). Hence these proteins were named TRF-1 interacting, ankyrin-related ADP-ribose polymerases, or TNKS.
  • TRF-1 is also a substrate of TNKS.
  • TNKS function as positive regulators of telomere length, permitting elongation of the telomeres by telomerase.
  • Another important substrate of TNKS is axin, a key regulator in the Wnt/beta- catenin signal transduction pathway.
  • the Wnt/beta-catenin pathway plays essential roles in embryonic development and adult tissue homeostasis and deregulation of this pathway has been linked to cancer.
  • Inhibitors of TNKS have been shown to result in efficient stabilization and increased levels of the axin-GSK3P complex protein which increases ⁇ - catenin phosphorylation and destruction.
  • the tankyrases are also proposed to have roles in the regulation of the mitotic spindle and in vesicle trafficking and they may also serve as scaffolds for proteins involved in various other cellular processes.
  • TGF beta transforming growth factor beta
  • the central role of TGF- ⁇ signaling is further highlighted by the development of a systemic fibrotic disease in mice with fibroblast-specific overexpression of constitutively active TGF- ⁇ receptor type 1 (Sonnylal et al., 2007).
  • pathologically activated canonical Wnt has been implicated in the pathogenesis of pulmonary-, renal-, dermal-, and liver-fibrosis as well as scarring after myocardial fibrosis following muscular dystrophy (Chilosi et al, 2003, Colwell et al, 2006, He et al, 2009 and 2010, Henderson et al, 2010, Konigshoff et al, 2008, Liu et al, 2009, Surendran et al., 2002 and Wei et al., 2011).
  • epithelial to mesenchymal transition requires epithelial integrin ⁇ 3 ⁇ 1 -mediated crosstalk between TGFpi and Wnt signaling pathway.
  • EMT has been shown to be inhibited by mutant ⁇ -catenin and tankyrase inhibitors (Ulsamer et al., 2012).
  • Tankyrase inhibitor dependent stabilization of Axin decreases in vivo fibrosis after bleomycin injury, suggesting that targeting axin levels (using tankyrase inhibitors) could attenuate fibrosis without blocking TGFpi homeostatic functions.
  • This invention relates to compounds of Formula (I)
  • R 2 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci-C 4 alkoxy;
  • R 4 is hydrogen or methyl
  • R 1 , R 2 and R 3 is not hydrogen
  • This invention relates to compounds of Formula (I), or pharmaceutically acceptable salts thereof.
  • This invention also relates to com ounds of Formula (I)(a):
  • R 2 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci-C 4 alkoxy;
  • R 4 is hydrogen or methyl; provided that at least one of R 1 , R 2 and R 3 is not hydrogen;
  • This invention also relates to com ounds of Formula (II):
  • R 2 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci-C 4 alkoxy;
  • R 1 , R 2 and R 3 is not hydrogen
  • This invention also relates to com ounds of Formula (II)(a):
  • R 2 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci-C 4 alkoxy;
  • R 1 , R 2 and R 3 is not hydrogen
  • This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R 1 is hydrogen; R 2 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci- C 4 alkoxy; R 4 is hydrogen; provided that at least one of R 2 and R 3 is not hydrogen;
  • This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R 2 is hydrogen; R 1 and R 3 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci- C 4 alkoxy; R 4 is hydrogen;
  • R 1 and R 3 are provided that at least one of R 1 and R 3 is not hydrogen;
  • This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R 3 is hydrogen; R 1 and R 2 are each independently hydrogen, halo, Ci-C 4 alkyl, or Ci- C 4 alkoxy; R 4 is hydrogen;
  • R 1 and R 2 are provided that at least one of R 1 and R 2 is not hydrogen;
  • This invention also relates to any one of the above compounds, wherein said substituted Ci-C 4 alkyl is Ci-C 4 haloalkyl; or a pharmaceutically acceptable salt thereof.
  • This invention also relates to any one of the above compounds, wherein said Ci- C 4 haloalkyl is CF 3 ; or a pharmaceutically acceptable salt thereof.
  • This invention also relates to compounds exemplified in the Experimental section.
  • This invention also relates to a method of treating cancer comprising administering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof.
  • This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), or any of the exemplified compounds, or their pharmaceutically acceptable salt thereof, for use as a medicament.
  • This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), or any of the exemplified compounds, or their pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cancer.
  • This invention also relates to a method of treating cancer comprising administering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof in a pharmaceutically acceptable composition.
  • This invention also relates to a method of treating cancer comprising coadministering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof and an antineoplastic agent.
  • salts of the present invention are pharmaceutically acceptable salts.
  • Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention.
  • Salts of the disclosed compounds containing a basic amine or other basic functional group may be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such as glucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citric acid or tartaric acid, amino acid, such as aspartic acid or glutamic acid, aromatic acid, such as be
  • Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates succinates, suberates, sebacates, fumarates, maleates, butyne-l,4-dioates, hexyne-l,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, phenylacetates, phenylpropionates, phenylbutrates, citrates, lactates, ⁇ -hydroxybutyrates, glycolates, tartrates mandelates
  • Salts of the disclosed compounds containing a carboxylic acid or other acidic functional group can be prepared by reacting with a suitable base.
  • a suitable base which affords a pharmaceutically acceptable cation, which includes alkali metal salts (especially sodium and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum salts and ammonium salts, as well as salts made from physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, ⁇ , ⁇ '-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2- hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, ⁇ , ⁇ '-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, choline, quinine,
  • salts which are not pharmaceutically acceptable, may be useful in the preparation of compounds of this invention and these should be considered to form a further aspect of the invention.
  • These salts such as oxalic or trifluoroacetate, while not in themselves pharmaceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable salts.
  • a compound of Formula (I), (I)(a), (II), or (II)(a)” or "the compound of Formula (I), (I)(a), (II), or (II)(a)” refers to one or more compounds according to Formula (I), (I)(a), (II), or (II)(a).
  • the compound of Formula (I), (I)(a), (II), or (II)(a) may exist in solid or liquid form. In the solid state, it may exist in crystalline or noncrystalline form, or as a mixture thereof.
  • pharmaceutically acceptable solvates may be formed for crystalline compounds wherein solvent molecules are incorporated into the crystalline lattice during crystallization.
  • Solvates may involve non-aqueous solvents such as, but not limited to, ethanol, isopropanol, DMSO, acetic acid, ethanolamine, or ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent incorporated into the crystalline lattice are typically referred to as "hydrates.” Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes all such solvates.
  • polymorphs may exhibit polymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs.”
  • the invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification.
  • polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.
  • the compound of Formula (I), (I)(a), (II), and (II)(a) or a salt thereof may exist in stereoisomeric forms (e.g., it contains one or more asymmetric carbon atoms).
  • the individual stereoisomers (enantiomers and diastereomers) and mixtures of these are included within the scope of the present invention.
  • the stereochemistry depicted in the compounds of Formula (I), (I)(a), (II), and (II)(a) is intended to convey the relative configuration of substituents on the cyclohexyl ring, including racemic mixtures of enantiomers, each individual enantiomer, and non-racemic mixtures of enantiomers.
  • a compound or salt of Formula (I), (I)(a), (II), or (II)(a) may exist in tautomeric forms other than that shown in the formula and these are also included within the scope of the present invention.
  • the present invention includes all combinations and subsets of the particular groups defined hereinabove.
  • the scope of the present invention includes mixtures of stereoisomers as well as purified enantiomers or enantiomerically/diastereomerically enriched mixtures. It is to be understood that the present invention includes all combinations and subsets of the particular groups defined hereinabove.
  • the subject invention also includes isotopically-labelled compounds, which are identical to those recited in Formula (I), (I)(a), (II), or (II)(a), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes that can be incorporated into compounds of the invention and pharmaceutically acceptable salts thereof include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulphur, fluorine, iodine, and chlorine, such as 2 H, 3 H, U C, 13 C, 14 C, 15 N, 17 0, 18 0, 31 P, 32 P, 35 S, 18 F, Compounds of the present invention and pharmaceutically acceptable salts of said compounds that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention.
  • Isotopically-labelled compounds of the present invention for example those into which radioactive isotopes such as 3 H, 14 C are incorporated, are useful in drug and/or substrate tissue distribution assays.
  • Tritiated, i.e., 3 H, and carbon- 14, i.e., 14 C, isotopes are particularly preferred for their ease of preparation and detectability.
  • n C and 18 F isotopes are particularly useful in PET (positron emission tomography), and 125 I isotopes are particularly useful in SPECT (single photon emission computerized tomography), all useful in brain imaging.
  • Isotopically labelled compounds of Formula (I), (I)(a), (II), or (II)(a) of this invention can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labelled reagent for a non-isotopically labelled reagent.
  • alkyl refers to a straight or branched chain alkyl, preferably having from one to twelve carbon atoms, which may be unsubstituted or substituted, saturated or unsaturated with multiple degrees of substitution, preferably 1 to 3. Suitable substituents are selected from the group consisting of: halogen, hydroxyl, methoxy, and ethoxy. Examples of “alkyl” as used herein include methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, t-butyl, isopentyl, n-pentyl, and the like, as well as substituted versions thereof, such as trifluoromethyl.
  • haloalkyl refers to an alkyl group, defined hereinabove, substituted with one or more, preferably one to three, halo substituents; examples of haloalkyl include trifluoromethyl.
  • cycloalkyl refers to an unsubstituted or substituted mono- or polycyclic non-aromatic saturated ring, which optionally includes an alkylene linker through which the cycloalkyl may be attached.
  • cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, as well as unsubstituted and substituted versions thereof.
  • alkoxy refers to the group -ORa, where Ra is Ci- C 4 alkyl or C3-C 7 cycloalkyl as defined above.
  • the term "optionally” means that the subsequently described event(s) may or may not occur, and includes both event(s) that occur and event(s) that do not occur.
  • the invention further provides a pharmaceutical composition (also referred to as pharmaceutical formulation) comprising a compound of Formula (I), (I)(a), (II), or (II)(a) or pharmaceutically acceptable salt thereof and one or more excipients (also referred to as carriers and/or diluents in the pharmaceutical arts).
  • a pharmaceutical composition also referred to as pharmaceutical formulation
  • excipients also referred to as carriers and/or diluents in the pharmaceutical arts.
  • excipients are acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof (i.e., the patient).
  • a process for the preparation of a pharmaceutical composition comprising mixing (or admixing) a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof with at least one excipient.
  • compositions may be in unit dose form containing a predetermined amount of active ingredient per unit dose.
  • a unit may contain a therapeutically effective dose of the compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof or a fraction of a therapeutically effective dose such that multiple unit dosage forms might be administered at a given time to achieve the desired therapeutically effective dose.
  • Preferred unit dosage formulations are those containing a daily dose or sub-dose, as herein above recited, or an appropriate fraction thereof, of an active ingredient.
  • such pharmaceutical compositions may be prepared by any of the methods well-known in the pharmacy art.
  • compositions may be adapted for administration by any appropriate route, for example, by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual, or transdermal), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal) routes.
  • oral including buccal or sublingual
  • rectal nasal
  • topical including buccal, sublingual, or transdermal
  • vaginal or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal) routes.
  • parenteral including subcutaneous, intramuscular, intravenous, or intradermal
  • compositions When adapted for oral administration, pharmaceutical compositions may be in discrete units such as tablets or capsules; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; oil-in-water liquid emulsions or water-in-oil liquid emulsions.
  • the compound or salt thereof of the invention or the pharmaceutical composition of the invention may also be incorporated into a candy, a wafer, and/or tongue tape formulation for administration as a "quick-dissolve" medicine.
  • the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • Powders or granules are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing, and coloring agents can also be present.
  • Capsules are made by preparing a powder mixture, as described above, and filling formed gelatin or non-gelatinous sheaths.
  • Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate, solid polyethylene glycol can be added to the powder mixture before the filling operation.
  • a disintegrating or solubilizing agent such as agar-agar, calcium carbonate, or sodium carbonate can also be added to improve the availability of the medicine when the capsule is ingested.
  • suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture.
  • Suitable binders include starch, gelatin, natural sugars, such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like.
  • Disintegrators include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.
  • Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant, and pressing into tablets.
  • a powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, and aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt, and/or an absorption agent such as bentonite, kaolin, or dicalcium phosphate.
  • a binder such as carboxymethylcellulose, and aliginate, gelatin, or polyvinyl pyrrolidone
  • a solution retardant such as paraffin
  • a resorption accelerator such as a quaternary salt
  • an absorption agent such as bentonite, kaolin, or dicalcium phosphate.
  • the powder mixture can be granulated by wetting a binder such as syrup, starch paste, acadia mucilage, or solutions of cellulosic or polymeric materials and forcing through a screen.
  • a binder such as syrup, starch paste, acadia mucilage, or solutions of cellulosic or polymeric materials
  • the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules.
  • the granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc, or mineral oil. The lubricated mixture is then compressed into tablets.
  • the compound or salt of the present invention can also be combined with a free-flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps.
  • a clear opaque protective coating consisting of a sealing coat of shellac, a coating of sugar, or polymeric material, and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different dosages.
  • Oral fluids such as solutions, syrups, and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of active ingredient.
  • Syrups can be prepared by dissolving the compound or salt thereof of the invention in a suitably flavoured aqueous solution, while elixirs are prepared through the use of a non- toxic alcoholic vehicle.
  • Suspensions can be formulated by dispersing the compound or salt of the invention in a non-toxic vehicle.
  • Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers, preservatives, flavor additives such as peppermint oil, natural sweeteners, saccharin, or other artificial sweeteners, and the like, can also be added.
  • dosage unit formulations for oral administration can be microencapsulated.
  • the formulation can also be prepared to prolong or sustain the release as, for example, by coating or embedding particulate material in polymers, wax, or the like.
  • the term "treatment” includes prophylaxis and refers to alleviating the specified condition, eliminating or reducing one or more symptoms of the condition, slowing or eliminating the progression of the condition, and preventing or delaying the reoccurrence of the condition in a previously afflicted or diagnosed patient or subject.
  • Prophylaxis or prevention or delay of disease onset is typically accomplished by administering a drug in the same or similar manner as one would to a patient with the developed disease or condition.
  • the present invention provides a potential treatment in a mammal, especially a human, suffering from disease conditions targeted by the present compounds.
  • Such treatment comprises the step of administering a therapeutically effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof to said mammal, particularly a human.
  • Treatment can also comprise the step of administering a therapeutically effective amount of a pharmaceutical composition containing a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof to said mammal, particularly a human.
  • the term "effective amount” means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought, for instance, by a researcher or clinician.
  • therapeutically effective amount means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder.
  • therapeutically effective amounts of a compound of Formula (I), (I)(a), (II), or (II)(a), as well as salts thereof may be administered as the raw chemical. Additionally, the active ingredient may be presented as a pharmaceutical composition.
  • a therapeutically effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof may be administered as the raw chemical, it is typically presented as the active ingredient of a pharmaceutical composition or formulation.
  • the precise therapeutically effective amount of a compound or salt thereof of the invention will depend on a number of factors, including, but not limited to, the age and weight of the subject (patient) being treated, the precise disorder requiring treatment and its severity, the nature of the pharmaceutical formulation/composition, and route of administration, and will ultimately be at the discretion of the attending physician or veterinarian.
  • a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof will be given for the treatment in the range of about 0.01 to 100 mg/kg body weight of recipient (patient, mammal) per day and more usually in the range of 0.1 to 10 mg/kg body weight per day.
  • Acceptable daily dosages may be from about 1 to about 1000 mg/day, and preferably from about 1 to about 100 mg/day. This amount may be given in a single dose per day or in a number (such as two, three, four, five, or more) of sub-doses per day such that the total daily dose is the same.
  • An effective amount of a salt thereof may be determined as a proportion of the effective amount of the compound of Formula (I), (I)(a), (II), or (II)(a) per se. Similar dosages should be appropriate for treatment (including prophylaxis) of the other conditions referred herein for treatment. In general, determination of appropriate dosing can be readily arrived at by one skilled in medicine or the pharmacy art.
  • compositions and methods provided herein can potentially be useful for the treatment of cancer including tumors such as skin, breast, brain, cervical carcinomas, testicular carcinomas, etc. More particularly, cancers that may be treated by the compositions and methods of the invention include, but are not limited to tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas.
  • tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas.
  • these compounds can potentially be used to treat: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar
  • bronchiolar carcinoma bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;
  • Gastrointestinal esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,
  • a compound of Formula (I), (I)(a), (II), or (II)(a) is administered for the treatment of cancer
  • co-administering and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of a TNKS1 or TNKS2 inhibiting compound, as described herein, and a further active ingredient or ingredients, known to be useful in the treatment of cancer, including chemotherapy and radiation treatment.
  • further active ingredient or ingredients, as used herein includes any compound or therapeutic agent known to or that demonstrates advantageous properties when administered to a patient in need of treatment for cancer.
  • the compounds are administered in a close time proximity to each other.
  • the compounds are administered in the same dosage form, e.g. one compound may be administered topically and another compound may be administered orally.
  • any anti-neoplastic agent that has activity versus a susceptible tumor being treated may be co-administered in the treatment of cancer in the present invention.
  • examples of such agents can be found in Cancer Principles and Practice of Oncology by V.T. Devita and S. Hellman (editors), 6 th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved.
  • Typical anti-neoplastic agents useful in the present invention include, but are not limited to, anti-microtubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclins, actinomycins and bleomycins; topoisomerase II inhibitors such as epipodophyllotoxins; antimetabolites such as purine and pyrimidine analogues and anti- folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents; and cell cycle signaling inhibitors.
  • anti-microtubule agents such as diterpenoids and vinca alkaloids
  • Examples of a further active ingredient or ingredients for use in combination or coadministered with the present TANKYRASE inhibiting compounds are chemotherapeutic agents.
  • Anti-microtubule or anti-mitotic agents are phase specific agents active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle.
  • anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids.
  • Diterpenoids which are derived from natural sources, are phase specific anti - cancer agents that operate at the G 2 /M phases of the cell cycle. It is believed that the diterpenoids stabilize the ⁇ -tubulin subunit of the microtubules by binding with this protein. Examples of diterpenoids include, but are not limited to, paclitaxel and its analog docetaxel.
  • Paclitaxel, 5P,20-epoxy-l,2a,4,7P,10p,13a-hexa-hydroxytax-l l-en-9-one 4,10- diacetate 2-benzoate 13 -ester with (2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. It is a member of the taxane family of terpenes. It was first isolated in 1971 by Wani et al. (Wani et al, J. Am. Chem, Soc, 93:2325.
  • Paclitaxel has been approved for clinical use in the treatment of refractory ovarian cancer in the United States (Markman et al., Yale Journal of Biology and Medicine, 64:583, 1991; McGuire et al, Ann. Intern, Med., 111 :273, 1989) and for the treatment of breast cancer (Holmes et al., J. Nat. Cancer Inst., 83: 1797, 1991). It is a potential candidate for the treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al., Sem. Oncol., 20:56, 1990).
  • the compound also shows potential for the treatment of polycystic kidney disease (Woo et. al., Nature, 368:750. 1994), lung cancer and malaria.
  • Treatment of patients with paclitaxel results in bone marrow suppression (multiple cell lineages, Ignoff, R.J. et. al, Cancer Chemotherapy Pocket Guide ⁇ 1998) related to the duration of dosing above a threshold concentration (50 nM) (Kearns, CM. et. al., Seminars in Oncology, 3(6) p.16- 23, 1995).
  • Docetaxel (2R,3S)-N-carboxy-3-phenylisoserine,N-tert-butyl ester, 13-ester with 5 -20-epoxy-l,2a,4,7 ,10 ,13a-hexahydroxytax-l l-en-9-one 4-acetate 2-benzoate, trihydrate; is commercially available as an injectable solution as TAXOTERE®.
  • Docetaxel is indicated for the treatment of breast cancer.
  • Docetaxel is a semisynthetic derivative of paclitaxel q.v., prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree. The dose limiting toxicity of docetaxel is neutropenia.
  • Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, and vinorelbine.
  • Vinblastine vincaleukoblastine sulfate
  • VELBAN® an injectable solution.
  • Myelosuppression is the dose limiting side effect of vinblastine.
  • Vincristine vincaleukoblastine, 22-oxo-, sulfate
  • ONCOVIN® an injectable solution.
  • Vincristine is indicated for the treatment of acute leukemias and has also found use in treatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.
  • Alopecia and neurologic effects are the most common side effect of vincristine and to a lesser extent myelosuppression and gastrointestinal mucositis.
  • Vinorelbine 3',4'-didehydro -4'-deoxy-C'-norvincaleukoblastine [R-(R*,R*)-2,3- dihydroxybutanedioate (l :2)(salt)], commercially available as an injectable solution of vinorelbine tartrate (NAVELBINE®), is a semisynthetic vinca alkaloid.
  • Vinorelbine is indicated as a single agent or in combination with other chemotherapeutic agents, such as cisplatin, in the treatment of various solid tumors, particularly non-small cell lung, advanced breast, and hormone refractory prostate cancers. Myelosuppression is the most common dose limiting side effect of vinorelbine.
  • Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA.
  • the platinum complexes enter tumor cells, undergo aquation, and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor.
  • Examples of platinum coordination complexes include, but are not limited to, cisplatin and carboplatin.
  • Cisplatin, cis-diamminedichloroplatinum, is commercially available as
  • Cisplatin is primarily indicated in the treatment of metastatic testicular and ovarian cancer and advanced bladder cancer.
  • the primary dose limiting side effects of cisplatin are nephrotoxicity, which may be controlled by hydration and diuresis, and ototoxicity.
  • Carboplatin, platinum, diammine [l,l-cyclobutane-dicarboxylate(2-)-0,0'], is commercially available as PARAPLATIN® as an injectable solution.
  • Carboplatin is primarily indicated in the first and second line treatment of advanced ovarian carcinoma. Bone marrow suppression is the dose limiting toxicity of carboplatin.
  • Alkylating agents are non-phase anti-cancer specific agents and strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophihc moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death.
  • alkylating agents include, but are not limited to, nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; and triazenes such as dacarbazine.
  • Cyclophosphamide 2-[bis(2-chloroethyl)amino]tetrahydro-2H-l,3,2- oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent or in combination with other chemotherapeutic agents, in the treatment of malignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea, vomiting and leukopenia are the most common dose limiting side effects of cyclophosphamide.
  • Melphalan 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as ALKERAN®. Melphalan is indicated for the palliative treatment of multiple myeloma and non-resectable epithelial carcinoma of the ovary. Bone marrow suppression is the most common dose limiting side effect of melphalan.
  • Chlorambucil 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Chlorambucil is indicated for the palliative treatment of chronic lymphatic leukemia, and malignant lymphomas such as lymphosarcoma, giant follicular lymphoma, and Hodgkin's disease. Bone marrow suppression is the most common dose limiting side effect of chlorambucil.
  • Busulfan 1 ,4-butanediol dimethanesulfonate, is commercially available as MYLERAN® TABLETS. Busulfan is indicated for the palliative treatment of chronic myelogenous leukemia. Bone marrow suppression is the most common dose limiting side effects of busulfan.
  • Carmustine 1, 3 -[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®.
  • Carmustine is indicated for the palliative treatment as a single agent or in combination with other agents for brain tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppression is the most common dose limiting side effects of carmustine.
  • dacarbazine 5 -(3, 3 -dimethyl- 1 -triazeno)-imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®.
  • dacarbazine is indicated for the treatment of metastatic malignant melanoma and in combination with other agents for the second line treatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dacarbazine.
  • Antibiotic anti-neoplastics are non-phase specific agents, which bind or intercalate with DNA. Typically, such action results in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids leading to cell death.
  • antibiotic anti-neoplastic agents include, but are not limited to, actinomycins such as dactinomycin, anthrocyclins such as daunorubicin and doxorubicin, and bleomycins.
  • Dactinomycin also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Dactinomycin is indicated for the treatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dactinomycin.
  • Daunorubicin (8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo- hexopyranosyl)oxy]-7,8,9, 10-tetrahydro-6,8, 11 -trihydroxy- 1 -methoxy-5, 12
  • naphthacenedione hydrochloride is commercially available as a liposomal injectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.
  • Daunorubicin is indicated for remission induction in the treatment of acute nonlymphocytic leukemia and advanced HIV associated Kaposi's sarcoma. Myelosuppression is the most common dose limiting side effect of daunorubicin.
  • Doxorubicin (8S, 10S)-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo- hexopyranosyl)oxy]-8-glycoloyl, 7,8,9, 10-tetrahydro-6,8, 11 -trihydroxy- 1 -methoxy-5, 12 naphthacenedione hydrochloride, is commercially available as an injectable form as RUBEX® or ADRIAMYCIN RDF®.
  • Doxorubicin is primarily indicated for the treatment of acute lymphoblastic leukemia and acute myeloblastic leukemia, but is also a useful component in the treatment of some solid tumors and lymphomas. Myelosuppression is the most common dose limiting side effect of doxorubicin.
  • Bleomycin a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus, is commercially available as BLENOXANE®. Bleomycin is indicated as a palliative treatment, as a single agent or in combination with other agents, of squamous cell carcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneous toxicities are the most common dose limiting side effects of bleomycin.
  • Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins.
  • Epipodophyllotoxins are phase specific anti-neoplastic agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and G 2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide and teniposide.
  • Etoposide 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-ethylidene-P-D- glucopyranoside]
  • VePESID® an injectable solution or capsules
  • VP- 16 Etoposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of testicular and non- small cell lung cancers. Myelosuppression is the most common side effect of etoposide. The incidence of leucopenia tends to be more severe than thrombocytopenia.
  • Teniposide 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-thenylidene-P-D- glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26.
  • Teniposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia in children. Myelosuppression is the most common dose limiting side effect of teniposide. Teniposide can induce both leucopenia and thrombocytopenia.
  • Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows.
  • Examples of antimetabolite anti-neoplastic agents include, but are not limited to, fluorouracil, methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.
  • 5 -Fluorouracil 5-fluoro-2,4- (1H,3H) pyrimidinedione
  • fluorouracil is commercially available as fluorouracil.
  • Administration of 5 -fluorouracil leads to inhibition of thymidylate synthesis and is also incorporated into both RNA and DNA. The result typically is cell death.
  • 5-fluorouracil is indicated as a single agent or in combination with other chemotherapy agents in the treatment of carcinomas of the breast, colon, rectum, stomach and pancreas. Myelosuppression and mucositis are dose limiting side effects of 5- fluorouracil.
  • Other fluoropyrimidine analogs include 5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridine monophosphate.
  • Cytarabine 4-amino-l-P-D-arabinofuranosyl-2 (lH)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. It is believed that cytarabine exhibits cell phase specificity at S-phase by inhibiting DNA chain elongation by terminal incorporation of cytarabine into the growing DNA chain. Cytarabine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Other cytidine analogs include 5-azacytidine and 2', 2 '-difluorodeoxy cytidine (gemcitabine). Cytarabine induces leucopenia, thrombocytopenia, and mucositis.
  • Mercaptopurine l,7-dihydro-6H-purine-6-thione monohydrate
  • PURINETHOL® is commercially available as PURINETHOL®.
  • Mercaptopurine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism.
  • Mercaptopurine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppression and gastrointestinal mucositis are expected side effects of mercaptopurine at high doses.
  • a useful mercaptopurine analog is azathioprine.
  • Thioguanine 2-amino-l,7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®.
  • Thioguanine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism.
  • Thioguanine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia.
  • Myelosuppression including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of thioguanine administration. However, gastrointestinal side effects occur and can be dose limiting.
  • Other purine analogs include pentostatin, erythrohydroxynonyladenine, fludarabine phosphate, and cladribine.
  • Gemcitabine 2'-deoxy-2', 2'-difluorocytidine monohydrochloride ( ⁇ -isomer), is commercially available as GEMZAR®.
  • Gemcitabine exhibits cell phase specificity at S- phase and by blocking progression of cells through the Gl/S boundary.
  • Gemcitabine is indicated in combination with cisplatin in the treatment of locally advanced non-small cell lung cancer and alone in the treatment of locally advanced pancreatic cancer.
  • Myelosuppression including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of gemcitabine administration.
  • Methotrexate N-[4[[(2,4-diamino-6-pteridinyl) methyljmethylamino] benzoyl]-L- glutamic acid, is commercially available as methotrexate sodium. Methotrexate exhibits cell phase effects specifically at S-phase by inhibiting DNA synthesis, repair and/or replication through the inhibition of dyhydrofolic acid reductase which is required for synthesis of purine nucleotides and thymidylate.
  • Methotrexate is indicated as a single agent or in combination with other chemotherapy agents in the treatment of choriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, and carcinomas of the breast, head, neck, ovary and bladder.
  • Myelosuppression (leucopenia, thrombocytopenia, and anemia) and mucositis are expected side effect of methotrexate administration.
  • Camptothecins including, camptothecin and camptothecin derivatives are available or under development as Topoisomerase I inhibitors. Camptothecins cytotoxic activity is believed to be related to its Topoisomerase I inhibitory activity.
  • camptothecins include, but are not limited to irinotecan, topotecan, and the various optical forms of 7-(4-methylpiperazino-methylene)-10,l l-ethylenedioxy-20-camptothecin described below.
  • Irinotecan is a derivative of camptothecin which binds, along with its active metabolite SN-38, to the topoisomerase I - DNA complex. It is believed that cytotoxicity occurs as a result of irrepairable double strand breaks caused by interaction of the topoisomerase I : DNA : irinotecan or SN-38 ternary complex with replication enzymes. Irinotecan is indicated for the treatment of metastatic cancer of the colon or rectum.
  • the dose limiting side effects of irinotecan HC1 are myelosuppression, including neutropenia, and GI effects, including diarrhea.
  • Topotecan HC1 (S)- 10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy- 1 H- pyrano[3',4',6,7]indolizino[l,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as the injectable solution HYCAMTIN®.
  • Topotecan is a derivative of camptothecin which binds to the topoisomerase I - DNA complex and prevents religation of singles strand breaks caused by Topoisomerase I in response to torsional strain of the DNA molecule.
  • Topotecan is indicated for second line treatment of metastatic carcinoma of the ovary and small cell lung cancer.
  • the dose limiting side effect of topotecan HC1 is myelosuppression, primarily neutropenia.
  • camptothecin derivative of formula A currently under development, including the racemic mixture (R,S) form as well as the R and S enantiomers:
  • Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer.
  • hormones and hormonal analogues useful in cancer treatment include, but are not limited to, adrenocorticosteroids such as prednisone and prednisolone which are useful in the treatment of malignant lymphoma and acute leukemia in children; aminoglutethimide and other aromatase inhibitors such as anastrozole, letrazole, vorazole, and exemestane useful in the treatment of adrenocortical carcinoma and hormone dependent breast carcinoma containing estrogen receptors; progestrins such as megestrol acetate useful in the treatment of hormone dependent breast cancer and endometrial carcinoma; estrogens, androgens, and anti-androgens such as flutamide, nilutamide, bicalutamide, cyproterone acetate and 5a-reductases
  • GnRH gonadotropin-releasing hormone
  • LH leutinizing hormone
  • FSH follicle stimulating hormone
  • Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change. As used herein this change is cell proliferation or differentiation.
  • Signal transduction inhibitors useful in the present invention include inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.
  • Protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth.
  • Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases.
  • Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain.
  • Receptor tyrosine kinases are involved in the regulation of cell growth and are generally termed growth factor receptors. Inappropriate or uncontrolled activation of many of these kinases, i.e.
  • aberrant kinase growth factor receptor activity for example by over- expression or mutation, has been shown to result in uncontrolled cell growth. Accordingly, the aberrant activity of such kinases has been linked to malignant tissue growth. Consequently, inhibitors of such kinases could provide cancer treatment methods.
  • Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor -I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene.
  • EGFr epidermal growth factor receptor
  • PDGFr platelet derived growth factor receptor
  • erbB2 erbB4
  • VEGFr vascular endothelial growth factor receptor
  • TIE-2 vascular endothelial growth factor receptor
  • TIE-2 t
  • inhibitors of growth receptors include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides.
  • Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in Kath, John C, Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al DDT Vol 2, No. 2 February 1997; and Lofts, F. J. et al, "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London.
  • Non-receptor tyrosine kinases which are not growth factor receptor kinases are termed nonreceptor tyrosine kinases.
  • Non-receptor tyrosine kinases useful in the present invention include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl.
  • Such nonreceptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S.
  • SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP.
  • SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T.E. (1995), Journal of Pharmacological and Toxicological Methods. 34(3) 125-32.
  • Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (raf), Mitogen or Extracellular Regulated Kinase (MEKs), and Extracellular Regulated Kinases (ERKs); and Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta).
  • IkB kinase family IK a, IK b
  • PKB family kinases AKT kinase family members
  • TGF beta receptor kinases TGF beta receptor kinases.
  • Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., (1999), Journal of Biochemistry, 126 (5) 799-803; Brodt, P, Samani, A., and Navab, R., (2000), Biochemical Pharmacology, 60, 1101-1107; Massague, J., Weis-Garcia, F., (1996), Cancer Surveys. 27:41-64; Philip, P.A., and Harris, A.L. (1995), Cancer Treatment and Research. 78: 3-27, Lackey, K. et al., (2000), Bioorganic and Medicinal Chemistry Letters, (10), 223-226; U.S. Patent No.
  • PI3 -kinase ATM, DNA-PK, and Ku are also useful in the present invention.
  • Such kinases are discussed in Abraham, R.T. (1996), Current Opinion in Immunology. 8 (3) 412-8; Canman, C.E., Lim, D.S. (1998), Oncogene 17 (25) 3301-3308; Jackson, S.P. (1997), International Journal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. et al, (2000), Cancer Res., 60(6), 1541-1545.
  • Myo-inositol signaling inhibitors such as phospho lipase C blockers and Myoinositol analogues.
  • signal inhibitors are described in Powis, G., and Kozikowski A., (1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workman and David Kerr, CRC press 1994, London.
  • Another group of signal transduction pathway inhibitors are inhibitors of Ras
  • Oncogene Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras, thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovsky, O.G., Rozados, V.R., Gervasoni, S.I. Matar, P. (2000), Journal of Biomedical Science. 7(4) 292-8; Ashby, M.N., (1998), Current Opinion in Lipidology. 9 (2) 99 - 102; and BioChim. Biophys. Acta, (1999) 1423(3): 19-30.
  • antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors.
  • This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases.
  • Imclone C225 EGFR specific antibody see Green, M.C. et al., Monoclonal Antibody Therapy for Solid Tumors, Cancer Treat.
  • Herceptin® erbB2 antibody see Tyrosine Kinase Signalling in Breast cancenerbB Family Receptor Tyrosine Kinases, Breast cancer Res., 2000, 2(3), 176-183
  • 2CB VEGFR2 specific antibody see Brekken, R.A. et al, Selective Inhibition of VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in mice, Cancer Res. (2000) 60, 5117-5124).
  • Non-receptor kinase angiogenesis inhibitors may also find use in the present invention.
  • Inhibitors of angiogenesis related VEGFR and TIE2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases).
  • Angiogenesis in general is linked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR have been shown to inhibit angiogenesis, primarily VEGF expression.
  • the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense.
  • non-receptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention.
  • anti-VEGF antibodies which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alpha v beta 3 ) that will inhibit angiogenesis; endostatin and angiostatin (non-RTK) may also prove useful in combination with the disclosed erb family inhibitors.
  • VEGFR the receptor tyrosine kinase
  • small molecule inhibitors of integrin alpha v beta 3
  • endostatin and angiostatin non-RTK
  • Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of Formula (I), (I)(a), (II), or (II)(a).
  • immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations.
  • the efficacy of immunologic approaches may be greatly enhanced through combined inhibition of erbB2/EGFR signaling pathways using a small molecule inhibitor. Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT et al. (2000), Cancer Res. 60: 3569-3576; and Chen Y, Hu D, Eling DJ, Robbins J, and Kipps TJ. (1998), Cancer Res.
  • Agents used in proapoptotic regimens may also be used in the combination of the present invention.
  • Members of the Bcl-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance.
  • EGF epidermal growth factor
  • Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle.
  • a family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle.
  • CDKs cyclin dependent kinases
  • Several inhibitors of cell cycle signalling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK6 and inhibitors for the same are described in, for instance, Rosania et al, Exp. Opin. Ther. Patents (2000), 10(2):215-230.
  • the cancer treatment method of the claimed invention includes the co-administration a compound of Formula (I), (I)(a), (II), or (II)(a) and/or a pharmaceutically acceptable salt, hydrate, solvate or pro-drug thereof and at least one anti- neoplastic agent, such as one selected from the group consisting of anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormonal analogues, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents, and cell cycle signaling inhibitors.
  • anti-microtubule agents platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormonal analogues, signal transduction pathway inhibitors, non-receptor
  • the compounds of this invention may be made by a variety of methods, including well-known standard synthetic methods. Illustrative general synthetic methods are set out below and then specific compounds of the invention are prepared in the working examples. The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. In all of the schemes described below, protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles of synthetic chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T.W. Green and P.G.M.
  • Compounds of Formula (I) may be prepared as illustrated in Scheme 1.
  • Substituted lH-benzo[d][l,3]oxazine-2,4-dione can be prepared by reacting the appropriately substituted 2-amino-benzoic acid IF with triphosgene in the presence of base, such as triethylamine, and in solvents such as THF.
  • Substituted 1H- benzo[d][l,3]oxazine-2,4-dione can then be coupled with trans-4-aminomethyl- cyclohexanecarboxylic acid to give 1A.
  • Treatment of amino acid 1A with triphosgene provides quinazoline acid IB.
  • the acid IB can be esterified by treatment with HC1 in MeOH to give ester 1C.
  • Alkylation of the ester 1C with appropriately substituted fluoro benzyl bromide provides fluoroquinazolinedione ID.
  • the fluoride group of ID can be replaced by cyano group using potassium cyanide to give quinazoline ester IE.
  • Hydrolysis of IE under basic condition provides the compounds of Formula (I).
  • 1C can be alkylated directly with the appropriately substituted 2-CN benzyl bromide to provide alkylated compound IE.
  • Scheme 1 Generic Synthesis of Quinazolinedione Carboxylic Acid
  • compound IC may be prepared as illustrated in Scheme 2.
  • the appropriately substituted 2-amino benzoic acid IF can be coupled with appropriately substituted amine to give amide IG.
  • IG can be cyclized by treatment with triphosgene to form compound IC.
  • Compound IC can be converted to the compounds of Formula (I) as described in Scheme 1.
  • the naming programs used are JChem for Excel, ACDLABs 11.0 Namebatch, ACD IUPAC or Chem Draw.
  • Example 1 trans-4-[( 1 - ⁇ [2-Cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -2,4-dioxo- 1 ,2,3 ,4- tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid
  • trans-4-(2,4-Dioxo- 1 ,4-dihydro-2H-quinazolin-3-ylmethyl)-cyclohexanecarboxylic acid (8 g, 26.49 mmol) was dissolved in HCl/MeOH (80 mL), and the resulted solution was heated to reflux for overnight. Solvent was removed to give the residue which was washed with water (200 mL) and extracted with DCM (20 mL X 2).
  • Example 4 trans-4- [( 1 - ⁇ [2-Cyano-3 -(trifluoromethyl)phenyl]methyl ⁇ -6-fluoro-5 -methyl- 2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
  • Example 7 4-[(l- ⁇ [2-Cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin- -yl)methyl]- 1 -methylcyclohexane- 1 -carboxylic acid
  • Example 8 trans-4- [( 1 - ⁇ [2-Cyano-3 -(trifluoromethyl)phenyl]methyl ⁇ -5 -methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
  • Example 11 trans-4-[(6-Chloro- 1 - ⁇ [2-cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
  • trans-methyl 4- (aminomethyl)cyclohexanecarboxylate (3 g, 17.52 mmol) was added, and the mixture was stirred at 20 °C. After LCMS showed that the reaction was complete, the mixture was dissolved in DCM (300 mL) and washed with water (200 mL X 3). The organic phase was dried over Na 2 S0 4 , filtered and concentrated under vacuum to give trans-methyl 4-((2- amino-5-chlorobenzamido)methyl)cyclohexanecarboxylate (4 g, 12.32 mmol, 70.3 % yield) as a solid.
  • Examples 12-13 were prepared using procedures analogous to those described in Example 8, starting from 4-(5-methyl-2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl) cyclohexanecarboxylic acid methyl ester (Intermediate 5) and the appropriately substituted benzyl bromide.
  • Table 1 Table 1:
  • Example 14 c 5-4-[(l- ⁇ [2-Cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid
  • Step 1 cis Methyl 4-(hydroxymethyl)cyclohexane-l-carboxylate
  • Step 3 cz ' s-Methyl 4-[(l- ⁇ [2-cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -5-methyl-2,4- dioxo-1, 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylate
  • Step 4 cz5-4-[(l- ⁇ [2-Cyano-3-(trifluoromethyl)phenyl]methyl ⁇ -5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid
  • Test compounds are plated at 0.1 ⁇ , diluted in 100% DMSO in low volume 384 well black polypropylene plates (NUNC 267461).
  • a positive control (XAV939) is added to column 18, to a final assay concentration of 20 uM, to define the maximal effect.
  • the ligand solution is prepared as 50 mM HEPES (pH 7.5), 10 mM MgCl 2 , 1 mM CHAPS, 1 mM DTT, 50 mM NaCl, and 1 nM ligand final concentrations.
  • TNKS 1 [or TNKS2] enzyme 5 nM final concentration is added to prepare the ligand/enzyme solution.
  • the assay is initiated upon the transfer 10 of the enzyme/ligand solution to each well of the compound plates using a Multidrop or similar instrument capable of accurately dispensing 10 ⁇ . Plates are then centrifuged for 1 minute at 500 x g. Assay plates are incubated for 1 to 2 hours at RT and then read on the Analyst GT using the PvhGr 505 filter set (ex at 485 nm, em at 530 nm, 505 dichroic) in the fluorescence polarization mode. Data generated are normalized to a maximal and no effect control. Potency of test molecules are reported as pIC 50 s (-log(IC 50 )).
  • Test compounds are plated at 0.1 uL diluted in 100% DMSO in low volume 384 well black polystyrene plates (Greiner 784076).
  • a positive control inhibitor (4-( ⁇ l-[(2-cyanophenyl)methyl]-2,4-dioxo-l ,2,3,4-tetrahydroquinazolin-3- yl ⁇ methyl)-N-(pyridin-4-yl)cyclohexane-l-carboxamide) is added to column 18, to a final assay concentration of 100 nM to define maximal effect.
  • the reaction buffer solution is prepared as 50 mM HEPES (pH 7.5) and 1 mM CHAPS.
  • TNKS 1 or TNKS2 enzyme 8 nM final concentration
  • the assay is initiated upon the transfer 5 uL of the 2X enzyme solution to each well of the compound plates using a Multidrop or similar instrument capable of accurately dispensing 5 uL. Plates are then incubated for 30 minutes.
  • a 2X substrate solution is prepared by adding 0.2 mM NAD, 50 nM GST-tev-Axin2 and lOmM MnC12 to the reaction buffer, and 5uL of this solution is added to the enzyme-test compound plate with a multidrop or similar instrument. Plates are then centrifuged for 1 minute at 500 x g, and incubated for 2 hours at RT. Finally, 2X detection mix is prepared, consisting of premade HTRF buffer
  • test molecules are reported as pICsoS (- logflCso)).
  • Examples 1-14 of the present invention were tested according to the assays described above and were found to be inhibitors of TANKYRASE with pIC 5 o >6 in one or both TNKS assays.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

This invention relates to the use of quinazolinediones for the modulation, notably the inhibition of the activity of tankyrases (TNKS1 and TNKS2). Suitably, the present invention relates to the use of quinazolinediones in the treatment of cancer, fibrosis and other hyperproliferative diseases through this mechanism.

Description

QUINAZOLINEDIONES AS TANKYRASE INHIBITORS
FIELD OF THE INVENTION
This invention relates to novel quinazolinediones which are inhibitors of tankyrases (TNKS1 and TNKS2), to pharmaceutical compositions containing them, and to their use in therapy for the treatment of cancers, fibrosis and other hyperproliferative diseases through this mechanism.
BACKGROUND OF THE INVENTION
The tankyrases (TNKS1 and TNKS2) belong to the poly ADP-ribose polymerase (PARP) family of enzymes and act via mono- or poly-ADP-ribosylation (parsylation) of substrate proteins. The tankyrases also contain the ANK domain, which contains 16-24 ankyrin repeats. The ANK domain interacts with a variety of proteins, including the telomeric protein Telomere Repeat binding Factor- 1 (TRF-1). Hence these proteins were named TRF-1 interacting, ankyrin-related ADP-ribose polymerases, or TNKS. TRF-1 is also a substrate of TNKS. Poly-ADP-ribosylation of TRF-1 inhibits the ability of TRF-1 to bind to telomeric DNA and leads to release of TRF-1 from the telomeres. The telomeric complex is opened up, allowing access to telomerase. Therefore TNKS function as positive regulators of telomere length, permitting elongation of the telomeres by telomerase. Another important substrate of TNKS is axin, a key regulator in the Wnt/beta- catenin signal transduction pathway. The Wnt/beta-catenin pathway plays essential roles in embryonic development and adult tissue homeostasis and deregulation of this pathway has been linked to cancer. Inhibitors of TNKS have been shown to result in efficient stabilization and increased levels of the axin-GSK3P complex protein which increases β- catenin phosphorylation and destruction. The tankyrases are also proposed to have roles in the regulation of the mitotic spindle and in vesicle trafficking and they may also serve as scaffolds for proteins involved in various other cellular processes.
The transforming growth factor beta (TGF beta) signaling pathway is a key mediator of fibroblast activation that drives the aberrant synthesis of extracellular matrix in fibrotic diseases (Wynn et al, 2008, Strieter et al, 2009, Verrecchia et al, 2007). The central role of TGF-β signaling is further highlighted by the development of a systemic fibrotic disease in mice with fibroblast-specific overexpression of constitutively active TGF-β receptor type 1 (Sonnylal et al., 2007). A link between TGF-β and the canonical Wnt pathway in fibrosis was recently made where tissue samples from human fibrotic disease showed enhanced expression of Wnt protein and decreased expression of the Wnt antagonist Dickkopf-1 (DK -1) (Akhmetshina et al., 2012). Moreover, the authors demonstrated that activation of the canonical Wnt pathway stimulated fibroblasts in vitro and induced fibrosis in vivo, suggesting that Wnt signaling is necessary for TGF-beta- mediated fibrosis and highlight a key role for the interaction of both pathways in the pathogenesis of fibrotic disease. Accumulating evidence indicates that increased activation of the canonical Wnt signaling might have an important role in fibrogenesis. Of particular interest, pathologically activated canonical Wnt has been implicated in the pathogenesis of pulmonary-, renal-, dermal-, and liver-fibrosis as well as scarring after myocardial fibrosis following muscular dystrophy (Chilosi et al, 2003, Colwell et al, 2006, He et al, 2009 and 2010, Henderson et al, 2010, Konigshoff et al, 2008, Liu et al, 2009, Surendran et al., 2002 and Wei et al., 2011). In addition to pulmonary fibrosis, epithelial to mesenchymal transition (EMT) requires epithelial integrin α3β1 -mediated crosstalk between TGFpi and Wnt signaling pathway. EMT has been shown to be inhibited by mutant β-catenin and tankyrase inhibitors (Ulsamer et al., 2012). Tankyrase inhibitor dependent stabilization of Axin decreases in vivo fibrosis after bleomycin injury, suggesting that targeting axin levels (using tankyrase inhibitors) could attenuate fibrosis without blocking TGFpi homeostatic functions.
All of these findings indicate that inhibition of tankyrase activity could have potential therapeutic benefit in the treatment of cancer, fibrosis and other hyperproliferative diseases through diverse modes of action.
SUMMARY OF THE INVENTION
This invention relates to compounds of Formula (I)
Figure imgf000004_0001
(I)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy;
R4 is hydrogen or methyl;
provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
This invention relates to compounds of Formula (I), or pharmaceutically acceptable salts thereof.
This invention also relates to com ounds of Formula (I)(a):
Figure imgf000004_0002
(0(a)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy;
R4 is hydrogen or methyl; provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
This invention also relates to com ounds of Formula (II):
Figure imgf000005_0001
(II)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy;
provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
This invention also relates to com ounds of Formula (II)(a):
Figure imgf000005_0002
(II)(a)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy;
provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R1 is hydrogen; R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci- C4alkoxy; R4 is hydrogen; provided that at least one of R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R2 is hydrogen; R1 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci- C4alkoxy; R4 is hydrogen;
provided that at least one of R1 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof. This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), wherein R3 is hydrogen; R1 and R2 are each independently hydrogen, halo, Ci-C4alkyl, or Ci- C4alkoxy; R4 is hydrogen;
provided that at least one of R1 and R2 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
This invention also relates to any one of the above compounds, wherein said substituted Ci-C4alkyl is Ci-C4haloalkyl; or a pharmaceutically acceptable salt thereof.
This invention also relates to any one of the above compounds, wherein said Ci- C4haloalkyl is CF3; or a pharmaceutically acceptable salt thereof.
This invention also relates to compounds exemplified in the Experimental section.
Specific compounds of this invention include:
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4- tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/?s-4-[(5-chloro- 1 - {[2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-fluoro-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6-fluoro-5-methyl-2,4- dioxo-1 , 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylic acid; trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -5 ,6-dimethyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
tran5-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methoxy-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4- tetrahydroquinazolin-3 -yl)methyl] - 1 -methylcyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -6-methoxy-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -6-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
tra/?s-4-[(6-chloro- 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
tra/75-4-({l-[(3-chloro-2-cyanophenyl)methyl]-5-methyl-2,4-dioxo-l, 2,3,4- tetrahydroquinazolin-3-yl}methyl)cyclohexane- 1 -carboxylic acid;
trans-4-{ { 1 -[(2-cyano-3-methylphenyl)methyl]-5-methyl-2,4-dioxo- 1 ,2,3,4- tetrahydroquinazolin-3-yl}methyl)cyclohexane- 1 -carboxylic acid; and
cis-4- [( 1 - { [2-cyano-3 -(trifluoromethyl)phenyl]methyl} -5 -methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
or pharmaceutically acceptable salts thereof.
This invention also relates to a method of treating cancer comprising administering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof.
This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), or any of the exemplified compounds, or their pharmaceutically acceptable salt thereof, for use as a medicament.
This invention also relates to compounds of Formula (I), (I)(a), (II), or (II)(a), or any of the exemplified compounds, or their pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cancer. This invention also relates to a method of treating cancer comprising administering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof in a pharmaceutically acceptable composition. This invention also relates to a method of treating cancer comprising coadministering to a human in need thereof an effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or a pharmaceutically acceptable salt thereof and an antineoplastic agent.
Typically, but not absolutely, the salts of the present invention are pharmaceutically acceptable salts. Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention. Salts of the disclosed compounds containing a basic amine or other basic functional group may be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such as glucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citric acid or tartaric acid, amino acid, such as aspartic acid or glutamic acid, aromatic acid, such as benzoic acid or cinnamic acid, sulfonic acid, such as p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid or the like. Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates succinates, suberates, sebacates, fumarates, maleates, butyne-l,4-dioates, hexyne-l,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, phenylacetates, phenylpropionates, phenylbutrates, citrates, lactates, γ-hydroxybutyrates, glycolates, tartrates mandelates, and sulfonates, such as xylenesulfonates, methanesulfonates, propanesulfonates, naphthalene- 1 -sulfonates and naphthalene -2-sulfonates.
Salts of the disclosed compounds containing a carboxylic acid or other acidic functional group can be prepared by reacting with a suitable base. Such a pharmaceutically acceptable salt may be made with a base which affords a pharmaceutically acceptable cation, which includes alkali metal salts (especially sodium and potassium), alkaline earth metal salts (especially calcium and magnesium), aluminum salts and ammonium salts, as well as salts made from physiologically acceptable organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, Ν,Ν'-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2- hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, Ν,Ν'-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, choline, quinine, quinoline, and basic amino acid such as lysine and arginine.
Other salts, which are not pharmaceutically acceptable, may be useful in the preparation of compounds of this invention and these should be considered to form a further aspect of the invention. These salts, such as oxalic or trifluoroacetate, while not in themselves pharmaceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable salts.
As used herein, the term "a compound of Formula (I), (I)(a), (II), or (II)(a)" or "the compound of Formula (I), (I)(a), (II), or (II)(a)" refers to one or more compounds according to Formula (I), (I)(a), (II), or (II)(a). The compound of Formula (I), (I)(a), (II), or (II)(a) may exist in solid or liquid form. In the solid state, it may exist in crystalline or noncrystalline form, or as a mixture thereof. The skilled artisan will appreciate that pharmaceutically acceptable solvates may be formed for crystalline compounds wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve non-aqueous solvents such as, but not limited to, ethanol, isopropanol, DMSO, acetic acid, ethanolamine, or ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes all such solvates.
The skilled artisan will further appreciate that certain compounds of the invention that exist in crystalline form, including the various solvates thereof, may exhibit polymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs." The invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. The skilled artisan will appreciate that different polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.
The compound of Formula (I), (I)(a), (II), and (II)(a) or a salt thereof may exist in stereoisomeric forms (e.g., it contains one or more asymmetric carbon atoms). The individual stereoisomers (enantiomers and diastereomers) and mixtures of these are included within the scope of the present invention. The stereochemistry depicted in the compounds of Formula (I), (I)(a), (II), and (II)(a) is intended to convey the relative configuration of substituents on the cyclohexyl ring, including racemic mixtures of enantiomers, each individual enantiomer, and non-racemic mixtures of enantiomers. Likewise, it is understood that a compound or salt of Formula (I), (I)(a), (II), or (II)(a) may exist in tautomeric forms other than that shown in the formula and these are also included within the scope of the present invention. It is to be understood that the present invention includes all combinations and subsets of the particular groups defined hereinabove. The scope of the present invention includes mixtures of stereoisomers as well as purified enantiomers or enantiomerically/diastereomerically enriched mixtures. It is to be understood that the present invention includes all combinations and subsets of the particular groups defined hereinabove.
The subject invention also includes isotopically-labelled compounds, which are identical to those recited in Formula (I), (I)(a), (II), or (II)(a), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention and pharmaceutically acceptable salts thereof include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulphur, fluorine, iodine, and chlorine, such as 2H, 3H, UC, 13C, 14C, 15N, 170, 180, 31P, 32P, 35S, 18F, Compounds of the present invention and pharmaceutically acceptable salts of said compounds that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as 3H, 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon- 14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. nC and 18F isotopes are particularly useful in PET (positron emission tomography), and 125I isotopes are particularly useful in SPECT (single photon emission computerized tomography), all useful in brain imaging. Further, substitution with heavier isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labelled compounds of Formula (I), (I)(a), (II), or (II)(a) of this invention can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labelled reagent for a non-isotopically labelled reagent.
DEFINITIONS
Terms are used within their accepted meanings. The following definitions are meant to clarify, but not limit, the terms defined.
As used herein, the term "alkyl" (or "alkylene") refers to a straight or branched chain alkyl, preferably having from one to twelve carbon atoms, which may be unsubstituted or substituted, saturated or unsaturated with multiple degrees of substitution, preferably 1 to 3. Suitable substituents are selected from the group consisting of: halogen, hydroxyl, methoxy, and ethoxy. Examples of "alkyl" as used herein include methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, t-butyl, isopentyl, n-pentyl, and the like, as well as substituted versions thereof, such as trifluoromethyl.
As used herein, the term "haloalkyl" refers to an alkyl group, defined hereinabove, substituted with one or more, preferably one to three, halo substituents; examples of haloalkyl include trifluoromethyl. As used herein, the term "cycloalkyl" refers to an unsubstituted or substituted mono- or polycyclic non-aromatic saturated ring, which optionally includes an alkylene linker through which the cycloalkyl may be attached. Exemplary "cycloalkyl" groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, as well as unsubstituted and substituted versions thereof.
As used herein, the term "alkoxy" refers to the group -ORa, where Ra is Ci- C4alkyl or C3-C7cycloalkyl as defined above.
As used herein, the term "optionally" means that the subsequently described event(s) may or may not occur, and includes both event(s) that occur and event(s) that do not occur.
PHARMACEUTICAL COMPOSITIONS
The invention further provides a pharmaceutical composition (also referred to as pharmaceutical formulation) comprising a compound of Formula (I), (I)(a), (II), or (II)(a) or pharmaceutically acceptable salt thereof and one or more excipients (also referred to as carriers and/or diluents in the pharmaceutical arts). The excipients are acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof (i.e., the patient).
In accordance with another aspect of the invention there is provided a process for the preparation of a pharmaceutical composition comprising mixing (or admixing) a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof with at least one excipient.
Pharmaceutical compositions may be in unit dose form containing a predetermined amount of active ingredient per unit dose. Such a unit may contain a therapeutically effective dose of the compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof or a fraction of a therapeutically effective dose such that multiple unit dosage forms might be administered at a given time to achieve the desired therapeutically effective dose. Preferred unit dosage formulations are those containing a daily dose or sub-dose, as herein above recited, or an appropriate fraction thereof, of an active ingredient. Furthermore, such pharmaceutical compositions may be prepared by any of the methods well-known in the pharmacy art. Pharmaceutical compositions may be adapted for administration by any appropriate route, for example, by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual, or transdermal), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal) routes. Such compositions may be prepared by any method known in the art of pharmacy, for example, by bringing into association the active ingredient with the excipient(s).
When adapted for oral administration, pharmaceutical compositions may be in discrete units such as tablets or capsules; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; oil-in-water liquid emulsions or water-in-oil liquid emulsions. The compound or salt thereof of the invention or the pharmaceutical composition of the invention may also be incorporated into a candy, a wafer, and/or tongue tape formulation for administration as a "quick-dissolve" medicine.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Powders or granules are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing, and coloring agents can also be present.
Capsules are made by preparing a powder mixture, as described above, and filling formed gelatin or non-gelatinous sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate, solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubilizing agent such as agar-agar, calcium carbonate, or sodium carbonate can also be added to improve the availability of the medicine when the capsule is ingested. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars, such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.
Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant, and pressing into tablets. A powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, and aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt, and/or an absorption agent such as bentonite, kaolin, or dicalcium phosphate. The powder mixture can be granulated by wetting a binder such as syrup, starch paste, acadia mucilage, or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc, or mineral oil. The lubricated mixture is then compressed into tablets. The compound or salt of the present invention can also be combined with a free-flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear opaque protective coating consisting of a sealing coat of shellac, a coating of sugar, or polymeric material, and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different dosages.
Oral fluids such as solutions, syrups, and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of active ingredient. Syrups can be prepared by dissolving the compound or salt thereof of the invention in a suitably flavoured aqueous solution, while elixirs are prepared through the use of a non- toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound or salt of the invention in a non-toxic vehicle. Solubilizers and emulsifiers, such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers, preservatives, flavor additives such as peppermint oil, natural sweeteners, saccharin, or other artificial sweeteners, and the like, can also be added. Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release as, for example, by coating or embedding particulate material in polymers, wax, or the like.
In the present invention, tablets and capsules are preferred for delivery of the pharmaceutical composition. As used herein, the term "treatment" includes prophylaxis and refers to alleviating the specified condition, eliminating or reducing one or more symptoms of the condition, slowing or eliminating the progression of the condition, and preventing or delaying the reoccurrence of the condition in a previously afflicted or diagnosed patient or subject. Prophylaxis (or prevention or delay of disease onset) is typically accomplished by administering a drug in the same or similar manner as one would to a patient with the developed disease or condition.
The present invention provides a potential treatment in a mammal, especially a human, suffering from disease conditions targeted by the present compounds. Such treatment comprises the step of administering a therapeutically effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof to said mammal, particularly a human. Treatment can also comprise the step of administering a therapeutically effective amount of a pharmaceutical composition containing a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof to said mammal, particularly a human.
As used herein, the term "effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought, for instance, by a researcher or clinician.
The term "therapeutically effective amount" means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function. For use in therapy, therapeutically effective amounts of a compound of Formula (I), (I)(a), (II), or (II)(a), as well as salts thereof, may be administered as the raw chemical. Additionally, the active ingredient may be presented as a pharmaceutical composition. While it is possible that, for use in therapy, a therapeutically effective amount of a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof may be administered as the raw chemical, it is typically presented as the active ingredient of a pharmaceutical composition or formulation. The precise therapeutically effective amount of a compound or salt thereof of the invention will depend on a number of factors, including, but not limited to, the age and weight of the subject (patient) being treated, the precise disorder requiring treatment and its severity, the nature of the pharmaceutical formulation/composition, and route of administration, and will ultimately be at the discretion of the attending physician or veterinarian. Typically, a compound of Formula (I), (I)(a), (II), or (II)(a) or salt thereof will be given for the treatment in the range of about 0.01 to 100 mg/kg body weight of recipient (patient, mammal) per day and more usually in the range of 0.1 to 10 mg/kg body weight per day. Acceptable daily dosages may be from about 1 to about 1000 mg/day, and preferably from about 1 to about 100 mg/day. This amount may be given in a single dose per day or in a number (such as two, three, four, five, or more) of sub-doses per day such that the total daily dose is the same. An effective amount of a salt thereof may be determined as a proportion of the effective amount of the compound of Formula (I), (I)(a), (II), or (II)(a) per se. Similar dosages should be appropriate for treatment (including prophylaxis) of the other conditions referred herein for treatment. In general, determination of appropriate dosing can be readily arrived at by one skilled in medicine or the pharmacy art.
The compositions and methods provided herein can potentially be useful for the treatment of cancer including tumors such as skin, breast, brain, cervical carcinomas, testicular carcinomas, etc. More particularly, cancers that may be treated by the compositions and methods of the invention include, but are not limited to tumor types such as astrocytic, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid carcinomas and sarcomas. More specifically, these compounds can potentially be used to treat: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar
(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, meduUoblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumor cervical dysplasia), ovaries (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplasia syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma. Thus, the term "cancerous cell" as provided herein, includes a cell afflicted by any one or related of the above identified conditions.
COMBINATIONS When a compound of Formula (I), (I)(a), (II), or (II)(a) is administered for the treatment of cancer, the term "co-administering" and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of a TNKS1 or TNKS2 inhibiting compound, as described herein, and a further active ingredient or ingredients, known to be useful in the treatment of cancer, including chemotherapy and radiation treatment. The term further active ingredient or ingredients, as used herein, includes any compound or therapeutic agent known to or that demonstrates advantageous properties when administered to a patient in need of treatment for cancer. Preferably, if the administration is not simultaneous, the compounds are administered in a close time proximity to each other. Furthermore, it does not matter if the compounds are administered in the same dosage form, e.g. one compound may be administered topically and another compound may be administered orally.
Typically, any anti-neoplastic agent that has activity versus a susceptible tumor being treated may be co-administered in the treatment of cancer in the present invention. Examples of such agents can be found in Cancer Principles and Practice of Oncology by V.T. Devita and S. Hellman (editors), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved. Typical anti-neoplastic agents useful in the present invention include, but are not limited to, anti-microtubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclins, actinomycins and bleomycins; topoisomerase II inhibitors such as epipodophyllotoxins; antimetabolites such as purine and pyrimidine analogues and anti- folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents; and cell cycle signaling inhibitors.
Examples of a further active ingredient or ingredients for use in combination or coadministered with the present TANKYRASE inhibiting compounds are chemotherapeutic agents.
Anti-microtubule or anti-mitotic agents are phase specific agents active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle. Examples of anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids.
Diterpenoids, which are derived from natural sources, are phase specific anti - cancer agents that operate at the G2/M phases of the cell cycle. It is believed that the diterpenoids stabilize the β-tubulin subunit of the microtubules by binding with this protein. Examples of diterpenoids include, but are not limited to, paclitaxel and its analog docetaxel.
Paclitaxel, 5P,20-epoxy-l,2a,4,7P,10p,13a-hexa-hydroxytax-l l-en-9-one 4,10- diacetate 2-benzoate 13 -ester with (2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. It is a member of the taxane family of terpenes. It was first isolated in 1971 by Wani et al. (Wani et al, J. Am. Chem, Soc, 93:2325. 1971), who characterized its structure by chemical and X-ray crystallographic methods. One mechanism for its activity relates to paclitaxel's capacity to bind tubulin, thereby inhibiting cancer cell growth. Schiff et al, Proc. Natl, Acad, Sci. USA, 77: 1561- 1565 (1980); Schiff et al, Nature, 277:665-667 (1979); Kumar, J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis and anticancer activity of some paclitaxel derivatives see: D. G. I. Kingston et al., Studies in Organic Chemistry vol. 26, entitled "New trends in Natural Products Chemistry 1986", Attaur-Rahman, P.W. Le Quesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.
Paclitaxel has been approved for clinical use in the treatment of refractory ovarian cancer in the United States (Markman et al., Yale Journal of Biology and Medicine, 64:583, 1991; McGuire et al, Ann. Intern, Med., 111 :273, 1989) and for the treatment of breast cancer (Holmes et al., J. Nat. Cancer Inst., 83: 1797, 1991). It is a potential candidate for the treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al., Sem. Oncol., 20:56, 1990). The compound also shows potential for the treatment of polycystic kidney disease (Woo et. al., Nature, 368:750. 1994), lung cancer and malaria. Treatment of patients with paclitaxel results in bone marrow suppression (multiple cell lineages, Ignoff, R.J. et. al, Cancer Chemotherapy Pocket Guide^ 1998) related to the duration of dosing above a threshold concentration (50 nM) (Kearns, CM. et. al., Seminars in Oncology, 3(6) p.16- 23, 1995).
Docetaxel, (2R,3S)-N-carboxy-3-phenylisoserine,N-tert-butyl ester, 13-ester with 5 -20-epoxy-l,2a,4,7 ,10 ,13a-hexahydroxytax-l l-en-9-one 4-acetate 2-benzoate, trihydrate; is commercially available as an injectable solution as TAXOTERE®. Docetaxel is indicated for the treatment of breast cancer. Docetaxel is a semisynthetic derivative of paclitaxel q.v., prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree. The dose limiting toxicity of docetaxel is neutropenia.
Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, and vinorelbine.
Vinblastine, vincaleukoblastine sulfate, is commercially available as VELBAN® as an injectable solution. Although, it has possible indication as a second line therapy of various solid tumors, it is primarily indicated in the treatment of testicular cancer and various lymphomas including Hodgkin's Disease; and lymphocytic and histiocytic lymphomas. Myelosuppression is the dose limiting side effect of vinblastine.
Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commercially available as ONCOVIN® as an injectable solution. Vincristine is indicated for the treatment of acute leukemias and has also found use in treatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas. Alopecia and neurologic effects are the most common side effect of vincristine and to a lesser extent myelosuppression and gastrointestinal mucositis.
Vinorelbine, 3',4'-didehydro -4'-deoxy-C'-norvincaleukoblastine [R-(R*,R*)-2,3- dihydroxybutanedioate (l :2)(salt)], commercially available as an injectable solution of vinorelbine tartrate (NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine is indicated as a single agent or in combination with other chemotherapeutic agents, such as cisplatin, in the treatment of various solid tumors, particularly non-small cell lung, advanced breast, and hormone refractory prostate cancers. Myelosuppression is the most common dose limiting side effect of vinorelbine. Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA. The platinum complexes enter tumor cells, undergo aquation, and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor. Examples of platinum coordination complexes include, but are not limited to, cisplatin and carboplatin. Cisplatin, cis-diamminedichloroplatinum, is commercially available as
PLATINOL® as an injectable solution. Cisplatin is primarily indicated in the treatment of metastatic testicular and ovarian cancer and advanced bladder cancer. The primary dose limiting side effects of cisplatin are nephrotoxicity, which may be controlled by hydration and diuresis, and ototoxicity. Carboplatin, platinum, diammine [l,l-cyclobutane-dicarboxylate(2-)-0,0'], is commercially available as PARAPLATIN® as an injectable solution. Carboplatin is primarily indicated in the first and second line treatment of advanced ovarian carcinoma. Bone marrow suppression is the dose limiting toxicity of carboplatin.
Alkylating agents are non-phase anti-cancer specific agents and strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophihc moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death. Examples of alkylating agents include, but are not limited to, nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; and triazenes such as dacarbazine.
Cyclophosphamide, 2-[bis(2-chloroethyl)amino]tetrahydro-2H-l,3,2- oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent or in combination with other chemotherapeutic agents, in the treatment of malignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea, vomiting and leukopenia are the most common dose limiting side effects of cyclophosphamide.
Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as ALKERAN®. Melphalan is indicated for the palliative treatment of multiple myeloma and non-resectable epithelial carcinoma of the ovary. Bone marrow suppression is the most common dose limiting side effect of melphalan.
Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Chlorambucil is indicated for the palliative treatment of chronic lymphatic leukemia, and malignant lymphomas such as lymphosarcoma, giant follicular lymphoma, and Hodgkin's disease. Bone marrow suppression is the most common dose limiting side effect of chlorambucil.
Busulfan, 1 ,4-butanediol dimethanesulfonate, is commercially available as MYLERAN® TABLETS. Busulfan is indicated for the palliative treatment of chronic myelogenous leukemia. Bone marrow suppression is the most common dose limiting side effects of busulfan.
Carmustine, 1, 3 -[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®. Carmustine is indicated for the palliative treatment as a single agent or in combination with other agents for brain tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppression is the most common dose limiting side effects of carmustine.
Dacarbazine, 5 -(3, 3 -dimethyl- 1 -triazeno)-imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®. Dacarbazine is indicated for the treatment of metastatic malignant melanoma and in combination with other agents for the second line treatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dacarbazine.
Antibiotic anti-neoplastics are non-phase specific agents, which bind or intercalate with DNA. Typically, such action results in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids leading to cell death. Examples of antibiotic anti-neoplastic agents include, but are not limited to, actinomycins such as dactinomycin, anthrocyclins such as daunorubicin and doxorubicin, and bleomycins.
Dactinomycin, also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Dactinomycin is indicated for the treatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dactinomycin.
Daunorubicin, (8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo- hexopyranosyl)oxy]-7,8,9, 10-tetrahydro-6,8, 11 -trihydroxy- 1 -methoxy-5, 12
naphthacenedione hydrochloride, is commercially available as a liposomal injectable form as DAUNOXOME® or as an injectable as CERUBIDINE®. Daunorubicin is indicated for remission induction in the treatment of acute nonlymphocytic leukemia and advanced HIV associated Kaposi's sarcoma. Myelosuppression is the most common dose limiting side effect of daunorubicin. Doxorubicin, (8S, 10S)-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo- hexopyranosyl)oxy]-8-glycoloyl, 7,8,9, 10-tetrahydro-6,8, 11 -trihydroxy- 1 -methoxy-5, 12 naphthacenedione hydrochloride, is commercially available as an injectable form as RUBEX® or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatment of acute lymphoblastic leukemia and acute myeloblastic leukemia, but is also a useful component in the treatment of some solid tumors and lymphomas. Myelosuppression is the most common dose limiting side effect of doxorubicin.
Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus, is commercially available as BLENOXANE®. Bleomycin is indicated as a palliative treatment, as a single agent or in combination with other agents, of squamous cell carcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneous toxicities are the most common dose limiting side effects of bleomycin.
Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins.
Epipodophyllotoxins are phase specific anti-neoplastic agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide and teniposide.
Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-ethylidene-P-D- glucopyranoside], is commercially available as an injectable solution or capsules as VePESID® and is commonly known as VP- 16. Etoposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of testicular and non- small cell lung cancers. Myelosuppression is the most common side effect of etoposide. The incidence of leucopenia tends to be more severe than thrombocytopenia. Teniposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-thenylidene-P-D- glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26. Teniposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia in children. Myelosuppression is the most common dose limiting side effect of teniposide. Teniposide can induce both leucopenia and thrombocytopenia.
Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows. Examples of antimetabolite anti-neoplastic agents include, but are not limited to, fluorouracil, methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.
5 -Fluorouracil, 5-fluoro-2,4- (1H,3H) pyrimidinedione, is commercially available as fluorouracil. Administration of 5 -fluorouracil leads to inhibition of thymidylate synthesis and is also incorporated into both RNA and DNA. The result typically is cell death. 5-fluorouracil is indicated as a single agent or in combination with other chemotherapy agents in the treatment of carcinomas of the breast, colon, rectum, stomach and pancreas. Myelosuppression and mucositis are dose limiting side effects of 5- fluorouracil. Other fluoropyrimidine analogs include 5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridine monophosphate.
Cytarabine, 4-amino-l-P-D-arabinofuranosyl-2 (lH)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. It is believed that cytarabine exhibits cell phase specificity at S-phase by inhibiting DNA chain elongation by terminal incorporation of cytarabine into the growing DNA chain. Cytarabine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Other cytidine analogs include 5-azacytidine and 2', 2 '-difluorodeoxy cytidine (gemcitabine). Cytarabine induces leucopenia, thrombocytopenia, and mucositis.
Mercaptopurine, l,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®. Mercaptopurine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Mercaptopurine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppression and gastrointestinal mucositis are expected side effects of mercaptopurine at high doses. A useful mercaptopurine analog is azathioprine.
Thioguanine, 2-amino-l,7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®. Thioguanine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Thioguanine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppression, including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of thioguanine administration. However, gastrointestinal side effects occur and can be dose limiting. Other purine analogs include pentostatin, erythrohydroxynonyladenine, fludarabine phosphate, and cladribine.
Gemcitabine, 2'-deoxy-2', 2'-difluorocytidine monohydrochloride (β-isomer), is commercially available as GEMZAR®. Gemcitabine exhibits cell phase specificity at S- phase and by blocking progression of cells through the Gl/S boundary. Gemcitabine is indicated in combination with cisplatin in the treatment of locally advanced non-small cell lung cancer and alone in the treatment of locally advanced pancreatic cancer. Myelosuppression, including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of gemcitabine administration.
Methotrexate, N-[4[[(2,4-diamino-6-pteridinyl) methyljmethylamino] benzoyl]-L- glutamic acid, is commercially available as methotrexate sodium. Methotrexate exhibits cell phase effects specifically at S-phase by inhibiting DNA synthesis, repair and/or replication through the inhibition of dyhydrofolic acid reductase which is required for synthesis of purine nucleotides and thymidylate. Methotrexate is indicated as a single agent or in combination with other chemotherapy agents in the treatment of choriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, and carcinomas of the breast, head, neck, ovary and bladder. Myelosuppression (leucopenia, thrombocytopenia, and anemia) and mucositis are expected side effect of methotrexate administration. Camptothecins, including, camptothecin and camptothecin derivatives are available or under development as Topoisomerase I inhibitors. Camptothecins cytotoxic activity is believed to be related to its Topoisomerase I inhibitory activity. Examples of camptothecins include, but are not limited to irinotecan, topotecan, and the various optical forms of 7-(4-methylpiperazino-methylene)-10,l l-ethylenedioxy-20-camptothecin described below.
Irinotecan HC1, (4S)-4,1 l-diethyl-4-hydroxy-9-[(4-piperidinopiperidino) carbonyloxy]-lH-pyrano[3 ',4',6,7]indolizino[l ,2-b]quinoline-3, 14(4H, 12H)-dione hydrochloride, is commercially available as the injectable solution CAMPTOSAR®.
Irinotecan is a derivative of camptothecin which binds, along with its active metabolite SN-38, to the topoisomerase I - DNA complex. It is believed that cytotoxicity occurs as a result of irrepairable double strand breaks caused by interaction of the topoisomerase I : DNA : irinotecan or SN-38 ternary complex with replication enzymes. Irinotecan is indicated for the treatment of metastatic cancer of the colon or rectum. The dose limiting side effects of irinotecan HC1 are myelosuppression, including neutropenia, and GI effects, including diarrhea. Topotecan HC1, (S)- 10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy- 1 H- pyrano[3',4',6,7]indolizino[l,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as the injectable solution HYCAMTIN®. Topotecan is a derivative of camptothecin which binds to the topoisomerase I - DNA complex and prevents religation of singles strand breaks caused by Topoisomerase I in response to torsional strain of the DNA molecule. Topotecan is indicated for second line treatment of metastatic carcinoma of the ovary and small cell lung cancer. The dose limiting side effect of topotecan HC1 is myelosuppression, primarily neutropenia.
Also of interest, is the camptothecin derivative of formula A, currently under development, including the racemic mixture (R,S) form as well as the R and S enantiomers:
Figure imgf000027_0001
known by the chemical name "7-(4-methylpiperazino-methylene)-10,l 1-ethylenedioxy- 20(R,S)-camptothecin (racemic mixture) or "7-(4-methylpiperazino-methylene)-10,l 1- ethylenedioxy-20(R)-camptothecin (R enantiomer) or "7-(4-methylpiperazino-methylene)- 10,l l-ethylenedioxy-20(S)-camptothecin (S enantiomer). Such compound as well as related compounds are described, including methods of making, in U.S. Patent Nos. 6,063,923; 5,342,947; 5,559,235; 5,491,237 and pending U.S. patent Application No. 08/977,217 filed November 24, 1997.
Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer. Examples of hormones and hormonal analogues useful in cancer treatment include, but are not limited to, adrenocorticosteroids such as prednisone and prednisolone which are useful in the treatment of malignant lymphoma and acute leukemia in children; aminoglutethimide and other aromatase inhibitors such as anastrozole, letrazole, vorazole, and exemestane useful in the treatment of adrenocortical carcinoma and hormone dependent breast carcinoma containing estrogen receptors; progestrins such as megestrol acetate useful in the treatment of hormone dependent breast cancer and endometrial carcinoma; estrogens, androgens, and anti-androgens such as flutamide, nilutamide, bicalutamide, cyproterone acetate and 5a-reductases such as finasteride and dutasteride, useful in the treatment of prostatic carcinoma and benign prostatic hypertrophy; anti- estrogens such as tamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene, as well as selective estrogen receptor modulators (SERMS) such those described in U.S. Patent Nos. 5,681,835, 5,877,219, and 6,207,716, useful in the treatment of hormone dependent breast carcinoma and other susceptible cancers; and gonadotropin-releasing hormone (GnRH) and analogues thereof which stimulate the release of leutinizing hormone (LH) and/or follicle stimulating hormone (FSH) for the treatment of prostatic carcinoma, for instance, LHRH agonists and antagonists such as goserelin acetate and luprolide.
Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change. As used herein this change is cell proliferation or differentiation. Signal transduction inhibitors useful in the present invention include inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.
Several protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth. Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases. Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain. Receptor tyrosine kinases are involved in the regulation of cell growth and are generally termed growth factor receptors. Inappropriate or uncontrolled activation of many of these kinases, i.e. aberrant kinase growth factor receptor activity, for example by over- expression or mutation, has been shown to result in uncontrolled cell growth. Accordingly, the aberrant activity of such kinases has been linked to malignant tissue growth. Consequently, inhibitors of such kinases could provide cancer treatment methods. Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor -I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene. Several inhibitors of growth receptors are under development and include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in Kath, John C, Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al DDT Vol 2, No. 2 February 1997; and Lofts, F. J. et al, "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London. Tyrosine kinases, which are not growth factor receptor kinases are termed nonreceptor tyrosine kinases. Non-receptor tyrosine kinases useful in the present invention, which are targets or potential targets of anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Such nonreceptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S. and Corey, S.J., (1999) Journal of Hematotherapy and Stem Cell Research 8 (5): 465 - 80; and Bolen, J.B., Brugge, J.S., (1997) Annual review of Immunology. 15: 371-404.
SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T.E. (1995), Journal of Pharmacological and Toxicological Methods. 34(3) 125-32.
Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (raf), Mitogen or Extracellular Regulated Kinase (MEKs), and Extracellular Regulated Kinases (ERKs); and Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta). IkB kinase family (IK a, IK b), PKB family kinases, AKT kinase family members, and TGF beta receptor kinases. Such Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., (1999), Journal of Biochemistry, 126 (5) 799-803; Brodt, P, Samani, A., and Navab, R., (2000), Biochemical Pharmacology, 60, 1101-1107; Massague, J., Weis-Garcia, F., (1996), Cancer Surveys. 27:41-64; Philip, P.A., and Harris, A.L. (1995), Cancer Treatment and Research. 78: 3-27, Lackey, K. et al., (2000), Bioorganic and Medicinal Chemistry Letters, (10), 223-226; U.S. Patent No. 6,268,391; and Martinez-Iacaci, L., et al, (2000), Int. J. Cancer, 88(1), 44- 52. Inhibitors of Phosphotidyl inositol-3 Kinase family members including blockers of
PI3 -kinase, ATM, DNA-PK, and Ku are also useful in the present invention. Such kinases are discussed in Abraham, R.T. (1996), Current Opinion in Immunology. 8 (3) 412-8; Canman, C.E., Lim, D.S. (1998), Oncogene 17 (25) 3301-3308; Jackson, S.P. (1997), International Journal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. et al, (2000), Cancer Res., 60(6), 1541-1545.
Also useful in the present invention are Myo-inositol signaling inhibitors such as phospho lipase C blockers and Myoinositol analogues. Such signal inhibitors are described in Powis, G., and Kozikowski A., (1994) New Molecular Targets for Cancer Chemotherapy ed., Paul Workman and David Kerr, CRC press 1994, London. Another group of signal transduction pathway inhibitors are inhibitors of Ras
Oncogene. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras, thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovsky, O.G., Rozados, V.R., Gervasoni, S.I. Matar, P. (2000), Journal of Biomedical Science. 7(4) 292-8; Ashby, M.N., (1998), Current Opinion in Lipidology. 9 (2) 99 - 102; and BioChim. Biophys. Acta, (1999) 1423(3): 19-30.
As mentioned above, antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases. For example Imclone C225 EGFR specific antibody (see Green, M.C. et al., Monoclonal Antibody Therapy for Solid Tumors, Cancer Treat. Rev., (2000), 26(4), 269-286); Herceptin® erbB2 antibody (see Tyrosine Kinase Signalling in Breast cancenerbB Family Receptor Tyrosine Kinases, Breast cancer Res., 2000, 2(3), 176-183); and 2CB VEGFR2 specific antibody (see Brekken, R.A. et al, Selective Inhibition of VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in mice, Cancer Res. (2000) 60, 5117-5124).
Non-receptor kinase angiogenesis inhibitors may also find use in the present invention. Inhibitors of angiogenesis related VEGFR and TIE2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases). Angiogenesis in general is linked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR have been shown to inhibit angiogenesis, primarily VEGF expression. Thus, the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense. Accordingly, non-receptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention. For example, anti-VEGF antibodies, which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alphav beta3) that will inhibit angiogenesis; endostatin and angiostatin (non-RTK) may also prove useful in combination with the disclosed erb family inhibitors. (See Bruns CJ et al (2000), Cancer Res., 60: 2926-2935; Schreiber AB, Winkler ME, and Derynck R. (1986), Science, 232: 1250-1253; Yen L et al. (2000), Oncogene 19: 3460-3469).
Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of Formula (I), (I)(a), (II), or (II)(a). There are a number of immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations. The efficacy of immunologic approaches may be greatly enhanced through combined inhibition of erbB2/EGFR signaling pathways using a small molecule inhibitor. Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT et al. (2000), Cancer Res. 60: 3569-3576; and Chen Y, Hu D, Eling DJ, Robbins J, and Kipps TJ. (1998), Cancer Res. 58: 1965-1971. Agents used in proapoptotic regimens (e.g., bcl-2 antisense oligonucleotides) may also be used in the combination of the present invention. Members of the Bcl-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance. Studies have shown that the epidermal growth factor (EGF) stimulates anti-apoptotic members of the bcl-2 family (i.e., mcl-1). Therefore, strategies designed to downregulate the expression of bcl-2 in tumors have demonstrated clinical benefit and are now in Phase II/III trials, namely Genta's G3139 bcl-2 antisense oligonucleotide. Such proapoptotic strategies using the antisense oligonucleotide strategy for bcl-2 are discussed in Water JS et al. (2000), J. Clin. Oncol. 18: 1812-1823; and Kitada S et al. (1994), Antisense Res. Dev. 4: 71-79.
Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle. A family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle. Several inhibitors of cell cycle signalling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK6 and inhibitors for the same are described in, for instance, Rosania et al, Exp. Opin. Ther. Patents (2000), 10(2):215-230.
In one embodiment, the cancer treatment method of the claimed invention includes the co-administration a compound of Formula (I), (I)(a), (II), or (II)(a) and/or a pharmaceutically acceptable salt, hydrate, solvate or pro-drug thereof and at least one anti- neoplastic agent, such as one selected from the group consisting of anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormonal analogues, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents, and cell cycle signaling inhibitors.
COMPOUNDS PREPARATION
The compounds of this invention may be made by a variety of methods, including well-known standard synthetic methods. Illustrative general synthetic methods are set out below and then specific compounds of the invention are prepared in the working examples. The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. In all of the schemes described below, protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles of synthetic chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T.W. Green and P.G.M. Wuts, (1991) Protecting Groups in Organic Synthesis, John Wiley & Sons, incorporated by reference with regard to protecting groups). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art. The selection of processes as well as the reaction conditions and order of their execution shall be consistent with the preparation of compounds of the present invention.
The synthesis of the compounds of the general Formula (I), (I)(a), (II), or (II)(a) and pharmaceutically acceptable derivatives and salts thereof may be accomplished as outlined below in Scheme 1-2 by those skilled in the art. In the following description, the groups are as defined above for compounds of Formula (I) unless otherwise indicated. Starting materials are commercially available or are made from commercially available starting materials using methods known to those skilled in the art.
Compounds of Formula (I) may be prepared as illustrated in Scheme 1. Substituted lH-benzo[d][l,3]oxazine-2,4-dione can be prepared by reacting the appropriately substituted 2-amino-benzoic acid IF with triphosgene in the presence of base, such as triethylamine, and in solvents such as THF. Substituted 1H- benzo[d][l,3]oxazine-2,4-dione can then be coupled with trans-4-aminomethyl- cyclohexanecarboxylic acid to give 1A. Treatment of amino acid 1A with triphosgene provides quinazoline acid IB. The acid IB can be esterified by treatment with HC1 in MeOH to give ester 1C. Alkylation of the ester 1C with appropriately substituted fluoro benzyl bromide provides fluoroquinazolinedione ID. The fluoride group of ID can be replaced by cyano group using potassium cyanide to give quinazoline ester IE. Hydrolysis of IE under basic condition provides the compounds of Formula (I). Alternatively, 1C can be alkylated directly with the appropriately substituted 2-CN benzyl bromide to provide alkylated compound IE. Scheme 1 : Generic Synthesis of Quinazolinedione Carboxylic Acid
Figure imgf000034_0001
Alternatively compound IC may be prepared as illustrated in Scheme 2. The appropriately substituted 2-amino benzoic acid IF can be coupled with appropriately substituted amine to give amide IG. IG can be cyclized by treatment with triphosgene to form compound IC. Compound IC can be converted to the compounds of Formula (I) as described in Scheme 1.
Scheme 2: Alternative synthesis of IC
Figure imgf000034_0002
Similarly, compounds of Formula (I)(a) may be prepared by analogous procedures to those depicted in Schemes 1 and 2 above employing the diastereomeric cyclohexyl moiety possessing the cis relationship between the carboxylic acid and the aminomethyl substituent. EXPERIMENTAL
The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention. While particular embodiments of the present invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention. Unless otherwise noted, reagents are commercially available or are prepared according to procedures in the literature. The symbols and conventions used in the descriptions of processes, schemes, and examples are consistent with those used in the contemporary scientific literature, for example, the Journal of the American Chemical Society or the Journal of Biological Chemistry.
In the Examples:
Chemical shifts are expressed in parts per million (ppm) units. Coupling constants (J) are in units of hertz (Hz). Splitting patterns describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), dd (double doublet), dt (double triplet), m (multiplet), br (broad).
Flash column chromatography was performed on silica gel.
The naming programs used are JChem for Excel, ACDLABs 11.0 Namebatch, ACD IUPAC or Chem Draw.
Abbreviations
AIBN azobisisobutyronitrile
(Boc)20 boc anhydride
nBuLi n-butyl lithium
CCU carbon tetrachloride
CH3CN acetonitrile
C02 carbon dioxide
Cs2C03 cesium carbonate
DCM dichloromethane DIEA diisopropylethylamine
DME dimethyl ether
DMF N,N-dimethylformamide
DMSO dimethylsulfoxide
EA ethyl acetate
EDCI 1 -ethyl-3 -(3 -dimethylaminopropyl)caarbodiimide
Et3N triethylamine
EtOAc ethyl acetate
h or hr hour(s)
H20 water
HC1 hydrochloric acid
HN03 nitric acid
HOBt hydroxybenzotriazole
HPLC high performance liquid chromatography
K2C03 potassium carbonate
KCN potassium cyanide
LCMS liquid chromatography-mass spectrometry
LDA lithium diisopropylamide
LiBH4 lithium borohydride
LiOH lithium hydroxide
Mel methyl iodide
MeOH methanol
N2 nitrogen gas
NaHC03 sodium bicarbonate
NBS N-bromosuccinimide
PBr3 phosphorus tribromide
PE petroleum ether
rt or r.t. room temperature
TEA triethylamine
THF tetrahydrofuran
Na2S203 sodium thiosulfate
Na2S04 sodium sulfate
Pd/C palladium on carbon Pd(PPh3)4 tetrakis(triphenylphosphine)palladium(0)
SOCl2 thionyl chloride
TLC thin layer chromotrography
Zn(CN)2 zinc cyanide
Preparation of Intermediates
Intermediate 1 : trans-M ethyl 4-(aminomethyl)cyclohexane-l-carboxylate
Figure imgf000037_0001
A solution of trans-4-aminomethyl-cyclohexanecarboxylic acid (10.0 g, 63.65 mmol) in HCl/MeOH (100 mL, 4 M) was stirred at rt overnight. LCMS showed that the reaction was finished. The mixture was concentrated under vacuum to give HCl salt of the target compound (11.0 g, 83.44% yield) as a white solid: 1H NMR (400 MHz, CD3OD) δ 3.64 (s, 3H), 2.82-2.81 (m, 2H), 2.38-2.33 (m, 1H), 2.08-2.04 (m, 2H), 1.92-1.88 (m, 2H), 1.67- 1.56 (m, 1H), 1.50-1.40 (m, 2H), 1.15-1.05 (m, 2H), ES-LCMS m/z: 172 (M+H).
Intermediate 2: 5-Methyl-2,4-dihydro- -3,l-benzoxazine-2,4-dione
Figure imgf000037_0002
To a suspension of 2-amino-6-methyl-benzoic acid (250 g, 1652 mmol) and triethylamine (346 mL, 2480 mmol) in THF (2000 mL) was added triphosgene (172 g, 578 mmol), the resulting solution was stirred at room temperature overnight, quenched with water (1000 mL) and extracted with ethyl acetate (1000 mL X 3). The organic layer was combined and concentrated. The residue was washed with methanol (50 mL X 3) to give 5-methyl-2,4- dihydro-lH-3,l-benzoxazine-2,4-dione (210 g, 72% yield) as a yellow solid: 1H NMR (400 MHz, DMSO-d6) δ 11.56 (br, 1H), 7.55-7.51 (m, 1H), 7.04-7.01 (m, 1H), 6.95 (d, J = 8.4 Hz, 1H), 2.57 (s, 3H); ES-LCMS m/z 178 (M+H).
Intermediate 3 : trans-4- { [(2-Amino-6-methylphenyl)formamido]methyl} cyclohexane- 1 - carboxylic acid
Figure imgf000038_0001
To a suspension of 5-methyl-2,4-dihydro-lH-3,l-benzoxazine-2,4-dione (5.0 g, 28.2 mmol) in ethanol (50 mL) and water (50 mL) was added trans-4-aminomethyl- cyclohexanecarboxylic acid (4.4 g, 28.2 mmol). The reaction mixture was stirred at 60 °C overnight. Then it was cooled to room temperature and concentrated to give crude trans- 4-[(2-amino-6-methyl-benzoylamino)-methyl]-cyclohexanecarboxylic acid (7.0 g, 86% yield) as a yellow solid: 1H NMR (400 MHz, CDC13) δ 7.55-7.51 (m, 1H), 6.93-6.89 (m, 1H), 6.48-6.44 (m, 1H), 3.18-3.15 (m, 2H), 2.22 (s, 3H), 2.12-2.11 (m, 1H), 1.96-1.93 (m, 2H), 1.85-1.82 (m, 2H), 1.57-1.47 (m, 1H), 1.35-1.30 (m, 2H), 1.02-0.93 (m, 2H); ES- LCMS m/z 291 (M+H).
Intermediate 4: tra/?5-4-[(5-Methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl] cyclohexane- 1 -carboxylic acid
Figure imgf000038_0002
To a suspension of trans-4- {[(2-amino-6- methylphenyl)formamido]methyl} cyclohexane- 1 -carboxylic acid (2.9 g, 10 mmol) and triethylamine (1.52 g, 15 mmol) in THF (30 mL) was added triphosgene (1.04 g, 3.5 mmol), the solution was stirred at 25 °C overnight, then quenched with water (30 mL). The mixture was concentrated under vacuum to remove THF. The resultant mixture was extracted with dichloromethane (30 mL X 3). The organic layer was combined, dried over anhydrous Na2S04 and filtered. The filtrate was concentrated to give trans-4-[(5- methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (2.0 g, 63% yield) as a yellow solid: 1H NMR (400 MHz, CD3OD) δ 7.53-7.49 (m, 1H), 7.08-7.04 (m, 2H), 3.90-3.89 (m, 2H), 2.78 (s, 3H), 2.28-2.24 (m, 1H), 2.04-1.79 (m, 5H), 1.42-1.36 (m, 2H), 1.23-1.13 (m, 2H); ES-LCMS m/z 317 (M+H).
Intermediate 5 : trans-M ethyl 4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl] cyclohexane- 1 -carboxylate
Figure imgf000039_0001
A suspension of tra/?5-4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (2.0 g, 6.3 mmol) in HCl/MeOH (20 mL) was stirred at room temperature for 24 h. The reaction mixture was concentrated to give crude trans-methyl 4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl] cyclohexane- 1 -carboxylate (1.3 g, 62% yield) as a yellow solid: 1H NMR (400 MHz, DMSO-d6) δ 7.49-7.45 (m, 1H), 7.09-6.93 (m, 2H), 3.75-3.73 (m, 2H), 2.51 (s, 3H), 2.76 (s, 3H), 2.29-2.23 (m, 1H), 1.91-1.87 (m, 2H), 1.74-1.65 (m, 3H), 1.29-1.22 (m, 2H), 1.13-1.03 (m, 2H). ES-LCMS m/z 331 (M+H).
Intermediate 6: 2-(Bromomethyl)-6-chlorobenzonitrile
Figure imgf000039_0002
A solution of 2-chloro-6-methyl-benzonitrile (1 g, 6.6 mmol), NBS (1.23 g, 6.93 mmol) and AIBN (0.33 g, 1.98 mmol) in CC14 (30 mL) was stirred at 80 °C overnight under N2.
After LC-MS analysis showed the starting material disappeared, solvent was removed in vacuo. The residue was dissolved in DCM (100 mL) and washed with Na2S203 (30 mL) and brine (30 mL). The organic layer was dried over Na2S04, filtered and concentrated to give 2-bromomethyl-6-chlorobenzonitrile (1.2 g, 80% yield), which was used in the next step without further purification: 1H NMR (400 MHz, CDC13) δ 7.52-7.45 (m, 2H), 7.23- 7.21 (m, 1H), 4.62 (s, 2H), ES-LCMS m/z: 231.8 (M+H).
Intermediate 7: 2-(Bromomethyl)-6-(trifluoromethyl)benzonitrile
Figure imgf000040_0001
Step 1 : 2-Fluoro-3-trifluoromethylbenzoic
Figure imgf000040_0002
To a solution of diisopropylamine (100 mL, 701 mmol) in THF (600 mL) was added nBuLi (268 mL, 670 mmol) at -30 °C. The mixture was stirred at 0 °C for 1 h. The resulting mixture (LDA) was used in the next step directly. To a solution of l-fluoro-2- (trifluoromethyl)benzene (100 g, 609 mmol) in THF (400 mL) was added LDA dropwise at -78 °C under N2. After addition, the mixture was stirred for 1 h at -78 °C. C02 was bubbled into the mixture for 50 min at -78 °C. The reaction was stirred for 1 h, warmed to room temperature, adjusted to pH 2 with 2 mol/L HCl, and extracted with EtOAc (800 mL X 3). The combined organic layers were washed with water and brine, dried over Na2S04, filtered, and concentrated to give 2-fluoro-3-(trifluoromethyl)benzoic acid (107 g, 84 % yield) as brown solid: 1H NMR (400 MHz, CDC13) δ 13.70 (br, 1H), 8.18-8.14 (m, 1H), 8.02-7.98 (m, 1H), 7.50 (t, J= 8.0 Hz, 1H); ES-LCMS m/z 209.0 (M+H). Step 2: Methyl 2-fluoro-3-(trifluoromethyl)benzoate
Figure imgf000041_0001
To a solution of 2-fluoro-3-trifluoromethylbenzoic acid (50 g, 240 mmol) in MeOH (250 mL) was added SOCl2 (70 g, 601 mmol) and DMF (5 mL) at room temperature. The solution was heated to refiux for 2 h. The mixture was concentrated in vacuo to give the crude product, which was washed with aqueous solution of NaHC03. The organic layer was separated, dried over Na2S04, filtered, and concentrated to give the desired product methyl 2-fluoro-3-(trifhioromethyl)benzoate (45 g, 84.2% yield): 1H NMR (400 MHz, CDC13) δ 8.16-8.12 (m, 1H), 7.79-7.77 (m, 1H), 7.34-7.26 (m, 1H), 3.96 (s, 3H); ES- LCMS m/z 222.9 (M+H).
Step 3 : Methyl 2-cyano-3-(trifluoromethyl)benzoate
Figure imgf000041_0002
To a solution of methyl 2-fluoro-3-trifluoromethylbenzoate (30 g, 135 mmol) in DMSO (150 mL) was added KCN (13.3 g, 202 mmol). The mixture was stirred at 60 °C for 3 h. The mixture was filtered, and the filtrate was concentrated to give the crude product. The crude product was purified by column (eluting with 5%~10% EtOAc in PE) to give the desired product methyl 2-cyano-3-(trifluoromethyl)benzoate (28 g, 90% yield): 1H NMR (400 MHz, CDC13) δ 8.32-8.30 (m, 1H), 8.01-7.99 (m, 1H), 7.83-7.80 (m, 1H), 4.05 (s, 3H); ES-LCMS m/z 230.1 (M+H). Step 4: 2-(Hydroxymethyl)-6-(trifluoromethyl)benzonitrile
Figure imgf000042_0001
To a solution of LiBH4 (3.3 g, 153 mmol) in THF (100 mL) was added methyl 2-cyano-3- (trifluoromethyl)benzoate (14 g, 61 mmol). The mixture was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to give the crude product, which was diluted with water, and extracted with EtOAc. The organic layer was separated, dried over Na2S04, filtered, and concentrated to give the crude product 2-(hydroxymethyl)-6- (trifluoromethyl)benzonitrile (10 g, 77% yield): 1H NMR (400 MHz, CDC13) δ 7.83-7.72 (m, 3H), 5.46 (s, 2H); ES-LCMS m/z 202.1 (M+H).
Step 5 : 2-(Bromomethyl)-6-(trifluoromethyl)benzonitrile
Figure imgf000042_0002
To a solution of 2-(hydroxymethyl)-6-(trifluoromethyl)benzonitrile (11 g, 53 mmol) in DCM (35 mL) was added PBr3 (21 g, 79 mmol) at room temperature. The solution was stirred at room temperature for 2 h. The mixture was concentrated in vacuo to give the crude product. The crude product was purified by column chromatography with PE: EtOAc (5: 1) to give the desired product 2-(bromomethyl)-6-(trifluoromethyl)benzonitrile (3.5 g, 25% yield): 1H NMR (400 MHz, CDC13) δ 7.79-7.71 (m, 3H), 4.70 (s, 3H); ES- LCMS m/z 264, 266 (M+2H). Intermediate 8: 2-(Bromomethyl)-6-methyl-benzonitrile
Figure imgf000043_0001
Step 1 : Methyl 2-cyano-3-methylbenzoate
Figure imgf000043_0002
A solution of methyl 2-bromo-3-methyl-benzoate (5 g, 21.8 mmol), Zn(CN)2 (10.26 g, 87.3 mmol) and Pd(PPh3)4 (1.51 g, 1.3 mmol) in DMF (60 mL) was stirred at 80 °C under N2 overnight. The solvent was removed in vacuo. The residue was dissolved in DCM (100 mL) and washed with H20 (50 mL). The organic layer was dried over Na2S04, filtered and concentrated to give the crude product, which was purified by column (eluting with 10% EtOAc in PE) to give methyl 2-cyano-3-methyl-benzoate (1.6 g, yield 43%) as a white solid: 1H NMR (400 MHz, CD3OD) δ 7.97-7.95 (m, 1H), 7.68-7.62 (m, 2H), 3.96 (s, 3H), 2.60 (s, 3H); ES-LCMS m/z: 176.1 (M+H).
Step 2: 2-(Hydroxymethyl)-6-methylbenzonitrile
Figure imgf000043_0003
To a solution of LiBH4 (0.62 g, 28.5 mmol) in MeOH (30 mL) was added methyl 2-cyano- 3-methyl-benzoate (1 g, 5.7 mmol). The resulting mixture was stirred at rt. After 1 hour, TLC analysis showed the starting material disappeared. The solvent was removed in vacuo. The residue was dissolved in DCM (100 mL) and washed with H20 (50 mL). The organic layer was dried over Na2S04, filtered and concentrated to give 2-(hydroxymethyl)- 6-methylbenzonitrile (0.9 g, 95% yield), which was used in the next step without further purification: 1H NMR (400 MHz, CDC13) δ 7.46 (d, J = 7.6 Hz, 1H), 7.39 (d, J = 7.6 Hz, 1H), 7.18-7.16 (m, 1H), 4.88 (s, 2H), 2.54 (s, 3H); ES-LCMS m/z: 148.1 (M+H). Step 3 : 2-Bromomethyl-6-methylbenzonitrile
Figure imgf000044_0001
To a mixture of 2-iodo- 1,3 -dimethyl-benzene (500 mg, 3.82 mmol) in CCI4 (10 mL) was added NBS (680 mg, 3.82 mmol) and AIBN (19 mg, 0.11 mmol). The reaction was stirred under N2 under reflux for 4 hours. Then the mixture was evaporated and the resulting residue was purified by column chromatography to give 2-bromomethyl-6-methyl- benzonitrile (400 mg, 50% yield) as a white solid, which was directly used in the next step: 1H NMR (400 MHz, CDC13) δ 7.47 (t, J = 2.8 Hz, 1H), 7.36 (d, J = 7.2 Hz, 1H), 7.28-7.27 (m, 1H), 4.64 (s, 2H), 2.57 (s, 3H); ES-LCMS m/z 210.0 (M+H).
Intermediate 9: trans-M ethyl 4-{[(6-amino-3-bromo-2-methylphenyl)formamido] methyl} cyclohexane- 1 -carboxylate
Figure imgf000044_0002
A mixture of 3-bromo-2-methyl-benzoic acid (20 g, 93 mmol) in HNO3 (20 mL), H2SO4 (100 mL) was stirred at room temperature for 3 hours. The mixture was poured into ice- water. The solid was collected via filtration, washed with water (3 x 10 mL) and dried to provide 3-bromo-2-methyl-6-nitrobenzoic acid as white solid (40 g, 40%> purity, 66.7% yield). 1H NMR (400 MHz, DMSO-/) δ 8.55 (d, J = 2.4 Hz, 1H), 8.46 (d, J = 2.4 Hz, 1H), 2.66 (s, 3H); ES-LCMS m/z 261.6 (M+2H).
To the mixture of 3-bromo-2-methyl-6-nitrobenzoic acid (9.4 g, 36 mmol) and Zn (23.5 g, 360 mmol) in MeOH (250 mL) was added NH4CI (19.3 g, 360 mmol) at room temperature, and the mixture was stirred at room temperature for 10 hr. The mixture was filtered, and the filtrate was concentrated to provide 6-amino-3-bromo-2-methylbenzoic acid as a brown solid (71% purity, 6 g with salt): 1H NMR (400 MHz, DMSO-/) δ 7.10 (d, J = 8.8 Hz, 1H), 6.42 (d, J= 8.8 Hz, 1H), 2.30 (s, 3H); ES-LCMS m/z 232 (M+2H).
A mixture of 6-amino-3-bromo-2-methylbenzoic acid (1 g, 4.35 mmol), trans-methyl 4- (aminomethyl)cyclohexanecarboxylate hydrochloride (900 mg, 4.35 mmol), DIEA (1.14 g, 8.8 mmol), EDCI (860 mg, 4.5 mmol) and HOBt (612 mg, 4.5 mmol) in DCM (20 mL) was stirred at room temperature for 16 hours. The mixture was washed with water (20 mL), and the organic phase was dried over anhydrous Na2S04, filtered and concentrated to provide trans-methyl 4-((6-amino-3 -bromo-2-methylbenzamido)methyl) cyclohexanecarboxylate, which was used in the next step without further purification (500 mg, 30% yield): 1H NMR (400 MHz, CD3OD) δ 7.27 (d, J = 8.8 Hz, 1H), 6.56 (d, J = 8.4 Hz, 1H), 3.67 (s, 3H), 3.23 (d, J = 6.8 Hz, 2H), 2.32 (s, 3H), 2.04-1.91 (m, 4H), 1.67-1.05 (m, 5H); ES-LCMS m/z 385 (M+2H).
Preparation of Compounds of the Invention
Example 1 : trans-4-[( 1 - { [2-Cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4- tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid
Figure imgf000045_0001
To a 200 mL pear-shaped flask containing trans-4-aminomethyl-cyclohexanecarboxylic acid (3.12 g, 20.0 mmol) was added isatoic anhydride (3.26 g, 20 mmol), ethanol (40 mL) and water (40 mL). The reaction mixture was allowed to stir at room temperature overnight. LCMS showed presence of desired intermediate. The mixture was filtered and the filter cake was washed with ethanol (10 mL), dried in vacuo to give trans-4-[(2-amino- benzoylamino)-methyl]-cyclohexanecarboxylic acid (4 g, 72.5 % yield): 1H NMR (400
MHz, CD3OD) δ 8.08-8.04 (m, 1H), 7.78-7.74 (m, 2H), 7.33-7.28 (m, 1H), 7.23-7.18 (m, 1H), 6.38 (s, 1H), 3.98-3.95 (d, J = 7.2 Hz, 2H), 2.30-2.25 (m, 1H), 1.98-1.95 (m, 2H), 1.62-1.60 (m, 2H), 1.09-1.03 (m, 2H); ES-LCMS m/z 277.2 (M+H).
To a stirred solution of tra/75-4-[(2-amino-benzoylamino)-methyl]-cyclohexanecarboxylic acid (0.55 g, 2.0 mmol) and triethylamine (0.2 g, 2.0 mmol) in THF (20 mL) was added trichloroacetyl chloride (0.39 g, 2.16 mmol) at 0 °C, then the mixture was stirred at r.t. for overnight, solvent was removed to give trans-4-{[2-(2,2,2-trichloro-acetylamino)- benzoylamino] -methyl} -eye lohexanecarboxylic acid, which was used in the next step without purification. (0.6 g, 79 % yield): 1H NMR (400 MHz, CDC13) δ 8.58-8.56 (m, 1H), 7.58-7.54 (m, 2H), 7.23-7.19 (m, 1H), 6.38 (s, 1H), 2.33-2.30 (m, 1H), 2.15-2.12 (m, 1H), 1.98-1.95 (m, 2H), 1.62-1.60 (m, 1H), 1.47-1.44 (m, 3H), 1.09-1.03 (m, 2H); ES- LCMS m/z 421.0 (M+H).
To a solution of tra/75-4-{[2-(2,2,2-trichloroacetylamino)-benzoylamino]-methyl}- cyclohexanecarboxylic acid (8.6 g, 20.4 mmol) in DMSO (100 mL) was added powdered sodium hydroxide (3.3 g, 81.6 mmol). The mixture was stirred at 80 °C for overnight. After cooling, the mixture was diluted with cold water (700 mL). The resultant mixture was acidified to pH 2 with 10% sulfuric acid, and extracted with DCM (50 mL X 3). The combined organic layers were dried over Na2S04, filtered and concentrated to afford tra/75-4-(2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl)-cyclohexanecarboxylic acid (3 g, 51.0 % yield): 1H NMR (400 MHz, CDC13) δ 8.02-7.99 (m, 1H), 7.64-7.60 (m, 1H), 7.23-7.21 (m, 1H), 7.19-7.13 (m, 1H), 3.88-3.87 (m, 2H), 2.24-2.20 (m, 1H), 1.99-1.96 (m, 2H), 1.84-1.79 (m, 3H), 1.38-1.35 (m, 2H), 1.15-1.10 (m, 2H); ES-LCMS m/z 303.2 (M+H).
trans-4-(2,4-Dioxo- 1 ,4-dihydro-2H-quinazolin-3-ylmethyl)-cyclohexanecarboxylic acid (8 g, 26.49 mmol) was dissolved in HCl/MeOH (80 mL), and the resulted solution was heated to reflux for overnight. Solvent was removed to give the residue which was washed with water (200 mL) and extracted with DCM (20 mL X 2). The combined organic layers were dried over Na2S04, filtered and concentrated to give methyl 4-(2,4-dioxo-l,4- dihydro-2H-quinazolin-3-ylmethyl)-cyclohexanecarboxylate (8 g, 89.8 % yield): ES- LCMS m/z 317.2 (Μ+Η).
A solution of tra/75-4-(2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl)- cyclohexanecarboxylic acid methyl ester (1.5 g, 4.75 mmol), l-bromomethyl-2-fluoro-3- trifluoromethyl-benzene (1.22 g, 4.75 mmol) and potassium carbonate (1.31 g, 9.49 mmol) in acetonitrile (100 mL) was heated to reflux for overnight, solvent was removed to give the residue which was purified by a flash column to give trans-methyl 4-[(l-{[2-fluoro-3- (trifluoromethyl)phenyl]methyl}-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (800 mg, 69.8 % yield): 1H NMR (400 MHz, CDCls) δ 8.20 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.55-7.50 (m, 2H), 7.26-7.21 (m, 2H), 7.19- 7.15 (m, 1H), 7.05-7.03 (m, 1H), 5.45 (s, 2H), 4.01 (d, J = 7.2 Hz, 2H), 3.68 (s, 3H), 2.25- 2.21 (m, 1H), 2.04-1.98 (m, 2H), 1.85-1.80 (m, 1H), 1.76-1.71 (m, 2H), 1.45-1.41 (m, 2H), 1.25-1.10 (m, 2H); ES-LCMS m/z 493.1 (M+H).
A solution of trans-methyl 4-[(l-{[2-fluoro-3-(trifluoromethyl)phenyl]methyl}-2,4-dioxo- l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (1 g, 2.03 mmol), potassium cyanide (400 mg, 6.09 mmol) in DMF (90 mL) was heated to 100 °C for overnight, solvent was removed to give the residue, which was purified by a flash column to give trans-methyl 4-[(l-{[2-cyano-3-(trif uoromethyl)phenyl]methyl}-2,4-dioxo- l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (600 mg, 69.8 % yield): 1H NMR (400 MHz, CDC13) δ 8.21 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.68-7.66 (m, 1H), 7.58-7.52 (m, 2H), 7.25-7.21 (m, 2H), 6.86-6.84 (d, J = 8.0 Hz, 1H), 5.59 (s, 2H), 3.96 (d, J= 7.6 Hz, 2H), 3.58 (s, 3H), 2.21-2.17 (m, 1H), 1.95-1.91 (m, 2H), 1.85-1.72 (m, 3H), 1.35-1.31 (m, 2H), 1.25-1.10 (m, 2H); ES-LCMS m/z 500.2 (M+H).
To a stirred solution of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-
2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (1.2 g, 2.4 mmol) in MeOH (20 mL) and water (20 mL) was added sodium hydroxide (0.48 g, 12.0 mmol). The mixture was stirred at r.t. for overnight. Saturated aqueous citric acid solution (100 mL) was added to the mixture, and the mixture was extracted with DCM (30 mL X 2). The combined organic layers were dried over Na2S04, filtered and concentrated to give trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3,4- tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (600 mg, 69.8 % yield): 1H NMR (400 MHz, CDC13) δ 8.20 (dd, J = 8.0, 1.6 Hz, IH), 7.70 (d, J = 8.0 Hz, IH), 7.63-7.56 (m, 2H), 7.27-7.24 (m, 2H), 6.90 (d, J = 8.4 Hz, IH), 5.60 (s, 2H), 3.96 (d, J = 7.2 Hz, 2H), 2.21-2.15 (m, IH), 1.98-1.94 (m, 2H), 1.90-1.73 (m, 3H), 1.38-1.34 (m, 2H), 1.21-1.10 (m, 2H); ES-LCMS m/z 486.1 (M+H).
Example 2: tra/?s-4-[(5-Chloro- 1 - {[2-cyano-3-(trifluoromethyl)phenyl]methyl} - 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid
Figure imgf000048_0001
A mixture of 2-amino-6-chloro-benzoic acid (2 g, 11.66 mmol), trans-4-aminomethyl- cyclohexanecarboxylic acid methyl ester (HC1 salt, 2.54 g, 12.24 mmol), HOBt (3.15 g, 23.32 mmol), EDCI (4.5 g, 23.32 mmol) and DIEA (4.5 g, 34.98 mmol) in THF (150 mL) was stirred at room temperature overnight. The reaction was quenched with water (25 mL) and diluted with DCM (25 mL). The organic layer was washed with brine (25 mL), dried over Na2S04, filtered and concentrated to give the residue, which was purified by a flash column to give tra/?5-4-[(2-amino-6-chloro-benzoylamino)-methyl]- cyclohexanecarboxylic acid methyl ester (3 g, 79% yield). 1H NMR (400 MHz, DMSO- d6) δ: 8.38 (s, IH), 6.98 (t, J = 8.0 Hz, IH), 6.62-6.56 (m, 2H), 5.12 (s, 2H), 3.56 (s, 3H), 3.05 (t, J = 7.0 Hz, 2H), 2.22-2.18 (m, IH), 1.90-1.79 (m, 4H), 1.52-1.50 (m, IH), 1.30- 1.26 (m, 2H), 0.98-0.95 (m, 2H); ES-LCMS m/z 326 (M+H). To a solution of tra/75-4-[(2-amino-6-chloro-benzoylamino)-methyl]- cyclohexanecarboxylic acid methyl ester (3 g, 9.26 mmol) and Et3N (2.8 mL, 18.52 mmol) in THF (150 mL) at 0 °C was added triphosgene (1.1 g, 3.70 mmol) in portions, the solution was stirred at room temperature overnight. The mixture was concentrated, and the residue was washed with H20 (5 mL) and MeOH (5 mL) to give trans-4-(5-chloro-2,4- dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl)-cyclohexanecarboxylic acid methyl ester (2.8 g, 80% yield). 1H NMR (400 MHz, DMSO-d6) δ: 11.51 (s, IH), 7.54 (t, J = 8.0 Hz, IH), 7.20 (d, J = 8.0 Hz, IH), 6.95 (d, J = 8.4 Hz, IH), 3.71 (d, J = 6.8 Hz, 2H), 3.55 (s, 3H), 2.27-2.23 (m, IH), 1.88-1.64 (m, 5H), 1.24-1.21 (m, 2H), 1.08-0.99 (m, 2H); ES- LCMS m/z 352 (M+H).
A mixture of tra/?5-4-(5-chloro-2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl)- cyclohexanecarboxylic acid methyl ester (350.5 mg, 1 mmol), l-bromomethyl-2-fluoro-3- trifluoromethyl-benzene (257 mg, 1 mmol), and Cs2C03 (352 mg, 2 mmol) in CH3CN (50 mL) was stirred at room temperature overnight. The mixture was diluted with DCM (25 mL), filtered, the filtrate was concentrated to give trans-methyl 4-[(l-{[2-fluoro-3- (trifluoromethyl)phenyl]methyl}-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (500 mg, 95% yield). 1H NMR (400 MHz, DMSO- d6) δ: 7.68 (t, J = 7.6 Hz, IH), 7.57 (t, J = 8.0 Hz, IH), 7.46 (t, J = 6.8 Hz, IH), 7.34-7.23 (m, 3H), 5.44 (s, 2H), 3.80 (d, J = 6.8 Hz, 2H), 3.31 (s, 3H), 2.27-2.23 (m, IH), 1.89-1.85 (m, 2H), 1.71-1.68 (m, 3H), 1.27-1.21 (m, 2H), 1.08-0.99 (m, 2H); ES-LCMS m/z 527 (M+H).
A solution of trans-methyl 4-[(l-{[2-fluoro-3-(trifluoromethyl)phenyl]methyl}-2,4-dioxo- l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (400 mg, 0.759 mmol), KCN (148 mg, 2.28 mmol) in DMSO (5 mL) was stirred in a microwave oven at 110 °C for 1 h. The mixture was diluted with H20 (5 mL) and extracted with DCM (15 mL). The organic layer was washed with brine (15 mL), dried over Na2S04, filtered, concentrated, and purified by prep-TLC to give trans-methyl 4-[(5-chloro-l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (100 mg, 24.7% yield). 1H NMR (400 MHz, CD3OD) δ: 7.63 (t, J= 7.6 Hz, IH), 7.55 (t, J = 8.4 Hz, IH), 7.37-7.25 (m, 3H), 7.15 (d, J = 8.4 Hz, IH), 5.53 (s, 2H), 3.97 (d, J = 7.2 Hz, 2H), 3.63 (s, 3H), 2.29-2.27 (m, IH), 1.99-1.80 (m, 5H), 1.42-1.30 (m, 2H), 1.20-1.09 (m, 2H); ES-LCMS m/z 534 (M+H).
To a stirred solution of trans-methyl 4-[(5-chloro-l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (150 mg, 0.28 mmol) in THF (10 mL) and water (10 mL) was added LiOH.H20 (71 mg, 1.69 mmol). The mixture was stirred at room temperature for 10 hours. The mixture was concentrated, acidified by 3 mol/L HCl to pH 2, and extracted with DCM (30 mL x 2). The organic layer was dried over Na2S04, filtered, concentrated, and purified by prep-HPLC to give tra/?5-4-[(5-chloro-l-{[2-cyano- 3-(trifluoromethyl)phenyl]methyl}-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (24.91 mg, 17.1% yield): 1H NMR (400 MHz, DMSO-d6) δ: 7.89 (d, J= 7.6 Hz, IH), 7.76 (t, J= 8.0 Hz, IH), 7.59-7.53 (m, 2H), 7.33 (d, J = 8.0 Hz, IH), 7.24 (d, J = 8.4 Hz, IH), 5.55 (s, 2H), 3.77 (s, 2H), 2.13-2.07 (m, IH), 1.86-1.83 (m, 2H), 1.70-1.68 (m, 3H), 1.23-1.17 (m, 2H), 1.08-0.98 (m, 2H); ES-LCMS m/z 520 (M+H).
Example 3 : tra/75-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-5-fluoro-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000050_0001
To a solution of 2-amino-6-fluorobenzoic acid (2.0 g, 12.89 mmol), EDCI (2.97 g, 15.47 mmol) and HOBt (2.09 g, 15.47 mmol) in DCM (30 mL) was added TEA (440 mg, 4.34 mmol), the resulted mixture was stirred at room temperature for 10 min. The resulting reaction mixture was then treated with trans-methyl 4- (aminomethyl)cyclohexanecarboxylate HCl salt (2.68 g, 12.89 mmol) and stirred at room temperature overnight. The mixture was then washed with brine (50 mL), dried over anhydrous Na2S04, filtered and concentrated in vacuo. The residue was purified by silica column to give trans-methyl 4-((2-amino-6-fluorobenzamido)methyl)cyclohexane carboxylate as the product (3.56 g, 89% yield): 1H NMR (400 MHz, CDC13) δ 7.25-7.03 (m, 1H), 6.64 (brs, 1H), 6.43-6.41 (m, 1H), 6.34-6.29 (m, 1H), 5.86 (brs, 2H), 3.64 (s, 3H), 3.29-3.26 (m, 2H), 2.27-2.20 (m, 1H), 2.02-1.90 (m, 2H), 1.89-1.86 (m, 2H), 1.60-1.54 (m, 1H), 1.47-13.7 (m, 2H), 1.07-0.97 (m, 2H); ES-LCMS m/z 309.2 (M+H). To a solution of trans-methyl 4-((2-amino-6-fluorobenzamido)methyl)cyclohexane carboxylate (1 g, 3.25 mmol) and TEA (0.66 g, 6.49 mmol) in THF (10 mL) at -20 °C was added a solution of triphosgene (0.39 g, 1.3 mmol) in THF dropwise. Then the reaction was allowed to warm to room temperature and stirred at this temperature overnight. The mixture was concentrated in vacuo. The residue was purified by silica column with EtOAc and PE as eluants to give trans-methyl 4-((5-f uoro-2,4-dioxo-l,2- dihydroquinazolin-3(4H)-yl)methyl)cyclohexanecarboxylate as a white solid (0.36g, 33% yield): 1H NMR (400 MHz, CDC13) δ 9.94 (s, 1H), 7.57-7.52 (m, 1H), 6.92-6.86 (m, 2H), 3.93 (d, J= 7.2 Hz, 2H), 3.64 (s, 3H), 2.26-2.25 (m, 1H), 2.04-1.98 (m, 2H), 1.88-1.75 (m, 3H), 1.40-1.25 (m, 2H), 1.17-1.13 (m, 2H); ES-LCMS m/z 335.1 (M+H).
A suspension of trans-methyl 4-((5-fluoro-2,4-dioxo-l,2-dihydroquinazolin-3(4H)- yl)methyl)cyclohexanecarboxylate (240 mg, 0.718 mmol) and Cs2C03 (0.476 g, 1.44 mmol) in acetonitrile (10 mL) was stirred at room temperature for 0.5 hr. Then 2- bromomethyl-6-trifluoromethyl-benzonitrile (199 mg, 0.75 mg) was added to the suspension. The reaction mixture was stirred overnight. The mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica column with EtOAc and PE as eluants to give trans-methyl 4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5-fluoro-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l -carboxylate as a white solid (0.32g, 86%> yield): 1H NMR (400 MHz, CDC13) δ 7.76-7.75 (m, 1H), 7.67-7.65 (m, 1H) 7.55-7.53 (m, 1H), 7.31-7.27 (m, 1H), 6.97-6.94 (m, 1H), 6.71-6.69 (m, 1H), 5.65 (s, 2H), 4.01 (d, J = 7.2 Hz, 2H), 3.65 (s, 3H), 2.26-2.23 (m, 1H), 2.04-2.00 (m, 2H), 1.99-1.79 (m, 3H), 1.42-1.30 (m, 2H), 1.29- 1.10 (m, 2H); ES-LCMS m/z 518.1 (M+H).
To a solution of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5- fluoro-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate
(160 mg, 0.309 mmol) in THF (5 mL) was added a solution of LiOH.H20 (39 mg, 0.928 mmol) in H20 (5 mL). The reaction mixture was stirred overnight. Solvent was removed in vacuo. The resulting aqueous solution was neutralized with 3 M HC1 to pH 2. The resulting white precipitate was collected by filtration to give tr<ms-4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5-fluoro-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (100 mg, 64% yield): 1H NMR (400 MHz, CDCls) δ 7.75 (d, J = 7.6 Hz, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.55-7.50 (m, 1H), 7.28-7.27 (m, 1H), 6.99-6.94 (m, 1H), 6.69 (d, J = 8.4 Hz, 1H), 5.64 (s, 2H), 4.00 (d, J = 7.2 Hz, 2H), 2.33-2.27 (m, 1H), 2.06-2.00 (m, 2H), 1.93-1.80 (m, 3H), 1.47-1.43 (m, 2H), 1.21- 1.08 (m, 2H); ES-LCMS m/z 504.1 (M+H).
Example 4: trans-4- [( 1 - { [2-Cyano-3 -(trifluoromethyl)phenyl]methyl} -6-fluoro-5 -methyl- 2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000052_0001
A mixture of 3-fluoro-2-methyl-benzoic acid (8.2 g, 41.2 mmol) and HNO3 (100 mL) was stirred at room temperature for 5 hour. LCMS showed the reaction was complete. The mixture was filtered, and the filter cake was collected to give 3-fluoro-2-methyl-6-nitro- benzoic acid as the desired product (8.1 g, 78.6 % yield). 1H NMR (400 MHz, CD3OD) δ 8.10 (dd, J = 9.2, 4.0 Hz, 1H), 7.34 (t, J = 9.2 Hz, 1H), 2.32 (s, 3H); ES-LCMS m/z 222 (M+23). To a mixture of 3-fluoro-2-methyl-6-nitro-benzoic acid (8.2 g, 41.2 mmol) in MeOH (80 mL) was added Pd/C (0.8 g) under N2 at room temperature. The mixture was stirred for 5 hours. LCMS showed the reaction was complete. The mixture was filtered, and the filtrate was concentrated to give 6-amino-3-fluoro-2-methyl-benzoic acid as the desired product (6.5 g, 93.4 % yield). 1H NMR (400 MHz, CD3OD) δ 6.93 (d, J = 8.8 Hz, 1H), 6.62 (m, 1H), 2.29 (s, 3H); ES-LCMS m/z 170 (M+H).
To a solution of 6-amino-3-fluoro-2-methyl-benzoic acid (3 g, 17.6 mmol) in DCM (50 mL) was added DIEA (6.8 g, 52.8 mmol). Then trans-4-aminomethyl- cyclohexanecarboxylic acid methyl ester (4.7 g, 22.9 mmol), EDCI (5.04 g, 26.4 mmol) and HOBt (3.59 g, 26.4 mmol) were added. The mixture was stirred at r.t. overnight. The mixture was extracted with DCM/H20 (80 mL). The organic layer was dried over Na2S04, filtered, and concentrated to give the crude product. Further purification via column chromatography gave tra/75-4-[(6-amino-3-fluoro-2-methyl-benzoylamino)- methylj-cyclohexane-carboxylic acid methyl ester as the desired product (4.5 g, 80.4 % yield). 1H NMR (400 MHz, DMSO-/) δ 11.31 (s, 1H), 7.46 (d, J = 9.2 Hz, 1H), 7.02 (d, J = 4.4 Hz, 1H), 3.72-3.70 (m, 2H), 3.31 (s, 3H), 2.48 (s, 3H), 2.23-2.11 (m, 1H), 1.87- 1.84 (m, 3H), 1.65-1.62 (m, 2H), 1.23-1.20(m, 2H), 1.17-1.04 (m, 2H); ES-LCMS m/z 323 (M+H).
To a solution of tra/75-4-[(6-amino-3-fluoro-2-methyl-benzoylamino)-methyl]- cyclohexane-carboxylic acid methyl ester (2.2 g, 6.83 mmol) in THF (30 mL) was added Et3N (1.3 g, 13.66 mmol). Then triphosgene (0.81 g, 2.73 mmol) in THF (5 mL) were added to the above mixture at 0 °C. The mixture was stirred at r.t. overnight. The mixture was extracted with DCM/H20 (50 mL), and the organic layer was dried over Na2S04, filtered, and concentrated to give the crude product. Further purification via column chromatography gave the desired product (2.27 g, 78.9 % yield). 1H NMR (400 MHz, CD3OD) δ 7.40 (d, J= 6.4 Hz, 1H), 7.08 (d, J= 9.2 Hz, 1H), 3.63 (s, 3H), 3.23-3.21 (m, 2H), 2.24 (s, 3H), 2.02-1.93 (m, 4H), 1.92-1.89 (m, 1H), 1.88-1.86 (m, 1H), 1.44-1.36 (m, 2H), 1.21-1.07 (m, 2H); ES-LCMS m/z 348.5 (M+H). To a solution of tra/75-4-(6-fluoro-5-methyl-2,4-dioxo-l,4-dihydro-2H-quinazolin-3- ylmethyl)-cyclohexanecarboxylic acid methyl ester (0.59 g, 1.71 mmol) in CH3CN (20 mL) was added K2CO3 (0.47 g, 3.42 mmol) and 2-bromomethyl-6- trifluoromethyl- benzonitrile (0.3 g, 1.14 mmol). The mixture was refluxed overnight. The mixture was extracted with EA/H20 (60 mL), the organic layer was dried over Na2S04, filtered, and concentrated to give the crude product, which was purified by column chromatography to give trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6-fluoro-5-methyl- 2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (290 mg, 31.8 % yield). 1H NMR (400 MHz, CDC13) δ 7.73 (d, J = 8.0 Hz, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.27-7.26 (m, 1H), 7.25-7.23 (m, 1H), 6.71-6.69 (m, 1H), 5.63 (s, 2H), 4.00 (d, J = 7.2 Hz, 2H), 3.64 (s, 3H), 2.74 (s, 3H), 2.21-2.16 (m, 1H), 2.01-1.98 (m, 2H), 1.81-1.78 (m, 2H), 1.42-1.41 (m, 2H), 1.39-1.38 (m, 1H), 1.27-1.24 (m, 2H); ES-LCMS m/z 532 (M+H).
To a stirred solution of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}- 6-fluoro-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l- carboxylate (250 mg, 0.47 mmol) in THF (5 mL) and water (5 mL) was added LiOH.H20 (131 mg, 2.35 mmol), then the mixture was stirred at r.t. for 10 hours. The mixture was concentrated, acidified by 3N HC1 to pH 2, and extracted with DCM (30 mL X 2). The combined organic layers were dried over Na2S04, filtered, concentrated, and purified by prepare HPLC to give tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6-fluoro- 5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (185 mg, 76.3 % yield): 1H NMR (400 MHz, CDC13) δ 7.73 (d, J = 8.0 Hz, 1H), 7.63 (d, J= 7.6 Hz, 1H), 7.28-7.25 (m, 1H), 7.24-7.23 (m, 1H), 6.70-6.68 (m, 1H), 5.62 (s, 2H), 3.99 (d, J= 7.2 Hz, 2H), 2.73 (s, 3H), 2.31-2.20 (m, 1H), 2.05-2.01 (m, 2H), 1.92-1.78 (m, 3H), 1.43-1.39 (m, 2H), 1.29-1.18 (m, 2H); ES-LCMS m/z 518 (M+H). Example 5: tra -4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyllmethyl|-5,6-dimethyl-2,4- dioxo-1, 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylic acid
Figure imgf000055_0001
A mixture of tra/75-4-[(6-amino-3-bromo-2-methyl-benzoylamino)-methyl]-cyclohexane carboxylic acid methyl ester (20 g, 52.3 mmol), methylboronic acid (3.77 g, 62.8 mmol), Pd(PPh3)2Cl2 (1.1 g, 1.57 mmol) and K2C03 (21.65 g, 157 mmol) in DME (300 mL) and H20 (10 mL) was stirred at 80 °C under N2 atmosphere for 16 hr. The mixture was concentrated, and DCM (300 mL) was added to the residue. The resulting mixture was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (EA/ PE = 21 1) to give the target product trans-4-[(6-amino-2,3- dimethyl-benzoylamino)-methyl]-cyclohexanecarboxylic acid methyl ester (2 g, 12% yield): 1H NMR (400 MHz, CDC13) δ 6.93 (d, J = 8.0 Hz, 1H), 6.48 (d, J = 8.4 Hz, 1H), 5.80 (br, 1H), 3.87 (br, 2H), 3.67 (s, 3H), 3.33 (t, J = 6.4 Hz, 2H), 2.31-2.25 (m, 1H), 2.20 (s, 3H), 2.15 (s, 3H), 2.15-2.01 (m, 2H), 1.93-1.89 (m, 2H), 1.61-1.51 (m, 1H), 1.46-1.43 (m, 2H), 1.11-1.04 (m, 2H); ES-LCMS m/z 319.2 (M+H).
To a cooled (0 °C) solution of tra/?5-4-[(6-amino-2,3-dimethyl-benzoylamino)-methyl]- cyclohexanecarboxylic acid methyl ester (1.6 g, 5.03 mmol) in THF (20 mL) was added a solution of triphosgene (0.67 g, 2.25 mmol) in THF (15 mL) dropwise. After the completion of addition, the mixture was stirred at room temperature for 1 hr, at which time the LCMS showed the completion of the reaction. The mixture was filtered, and the solid was collected to give tra/?5-4-(5,6-dimethyl-2,4-dioxo-l,4-dihydro-2H-quinazolin-3- ylmethyl)-cyclohexanecarboxylic acid methyl ester (1 g, 57.8% yield): 1H NMR (400 MHz, CDC13) δ 8.80 (br, 1H), 7.35 (d, J= 8.4 Hz, 1H), 6.77 (d, J = 8.4 Hz, 1H), 3.92 (d, J = 7.2 Hz, 2H), 3.64 (s, 3H), 2.75 (s, 3H), 2.33 (s, 3H), 2.29-2.23 (m, 1H), 2.01-1.97 (m, 2H), 1.90-1.80 (m, 3H), 1.44-1.34 (m, 2H), 1.20-1.10 (m, 2H); ES-LCMS m/z 345.2 (M+H).
A mixture of trans-4-(5, 6-dimethyl-2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl)- cyclohexanecarboxylic acid methyl ester (260 mg, 0.756 mmol), 2-bromomethyl-6- trifluoromethyl-benzonitrile (200 mg, 0.76 mmol) and K2CO3 (209 mg, 1.51 mmol) in DMF (20 mL) was heated to 80 °C overnight. The reaction mixture was concentrated and the residue was purified by column chromatography (MeOH/ DCM = 1/ 30) to give trans- methyl 4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -5 ,6-dimethyl-2,4-dioxo- l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (380 mg, 95% yield): 1H NMR (400 MHz, CDC13) δ 7.65 (d, J= 7.6 Hz, 1H), 7.53 (t, J = 8.0 Hz, 1H), 7.26 (d, J = 8.4 Hz, 1H), 7.20 (s, 1H), 6.57 (d, J = 8.8 Hz, 1H), 5.57 (s, 2H), 3.94 (d, J= 7.2 Hz, 2H), 3.58 (s, 3H), 2.72 (s, 3H), 2.26 (s, 3H), 2.21-2.18 (m, 1H), 1.96-1.92 (m, 2H), 1.85-1.80 (m, 1H), 1.74 (d, J = 13.6 Hz, 2H), 1.39-1.32 (m, 2H), 1.15-1.05 (m, 2H); ES-LCMS m/z 528.3 (M+ ).
A mixture of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5,6- dimethyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (380 mg, 0.72 mmol) and NaOH (58 mg, 1.44 mmol) in H20 (30 mL) was stirred at 50 °C for 4 hr. The mixture was neutralized with 2 N HC1 and concentrated. The residue was purified by chromatography (MeOH/DCM =1/30) to give tra/?5-4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5,6-dimethyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (300 mg, 81 % yield): 1H NMR (400 MHz, CD3OD) δ 7.82 (d, J= 7.6 Hz, 1H), 7.73 (t, J= 8.0 Hz, 1H), 7.41 (d, J = 8.4 Hz, 2H), 6.87 (d, J= 8.4 Hz, 1H), 5.65 (s, 2H), 3.96 (d, J = 6.8 Hz, 2H), 2.77 (s, 3H), 2.32 (s, 3H), 2.25- 2.19 (m, 1H), 2.00-1.97 (m, 2H), 1.81-1.78 (m, 2H), 1.41-1.31 (m, 2H), 1.30-1.21 (m, 1H), 1.18-1.13 (m, 2H); ES-LCMS m/z 514.2 (M+H).
Example 6 : tra/75-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-5-methoxy-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000057_0001
A mixture of 2-amino-6-methoxy-benzoic acid (1 g, 6 mmol), trans-4-aminomethyl- cyclohexanecarboxylic acid methyl ester (HC1 salt, 1.3 g, 6.3 mmol), HOBT (1.62 g, 12 mmol), EDCI (2.3 g, 12 mmol) and DIEA (2.3 g, 18 mmol) in THF (100 mL) was stirred at room temperature overnight. The reaction was quenched by water (25 mL) and diluted with DCM (25 mL). The organic layer was washed with brine (25 mL), dried over Na2S04, filtered and concentrated to give the residue, which was purified by a flash silica column to give tra/?5-4-[(2-amino-6-methoxy-benzoylamino)-methyl]- cyclohexanecarboxylic acid methyl ester (600 mg, 31.6% yield): 1H NMR (400 MHz, CD3OD) δ: 7.08 (t, J = 8.0 Hz, IH), 6.39 (dd, J = 8.4 Hz, 0.8 Hz, IH), 6.31 (dd, J = 8.4 Hz, 0.8 Hz, IH), 3.84 (s, 3H), 3.65 (s, 3H), 3.23 (d, J = 6.8 Hz, 2H), 2.32-3.28 (m, IH), 2.05-1.92 (m, 4H), 1.61-1.60 (m, IH), 1.50-1.39 (m, 2H), 1.14-1.07 (m, 2H); ES-LCMS m/z 321 (M+H).
To a solution of tra/?5-4-[(2-amino-6-methoxy-benzoylamino)-methyl]- cyclohexanecarboxylic acid methyl ester (600 mg, 1.88 mmol) and Et3N (0.53 mL, 3.76 mmol) in THF (50 mL) at 0 °C was added triphosgene (223 mg, 0.76 mmol), then the reaction mixture was stirred at room temperature overnight. The mixture was concentrated, and the residue was washed with water (25 mL) and cold MeOH (25 mL) to give trans-methyl 4-[(5-methoxy-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (500 mg, 77% yield): 1H NMR (400 MHz, DMSO- d6) δ 11.25 (br, IH), 7.45 (t, J = 8.0 Hz, IH), 7.01-6.94 (m, 2H), 3.72 (d, J = 6.8 Hz, 2H), 3.55 (s, 3H), 2.65 (s, 3H), 2.28-2.20 (m, 1H), 1.88-1.85 (m, 2H), 1.84-1.67 (m, 3H), 1.27- 1.16 (m, 2H), 1.05-0.90 (m, 2H); ES-LCMS m/z 347 (M+H).
A mixture of l-bromomethyl-2-fluoro-3-trifluoromethyl-benzene (650 mg, 1.88 mmol), trans-methyl 4-[(5-methoxy-2,4-dioxo-l ,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (506 mg, 2.06 mmol), and CS2CO3 (1.23 g, 2.76 mmol) in CH3CN (50 mL) was stirred at room temperature overnight. The mixture was filtered and concentrated to give trans-methyl 4-[(l-{[2-fluoro-3- (trifluoromethyl)phenyl]methyl}-5-methoxy-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (800 mg, 81.6% yield): 1H NMR (400 MHz, DMSO-d6) δ: 7.75-7.72 (m, 1H), 7.67 (t, J = 7.2 Hz, 1H), 7.52-7.26 (m, 2H), 6.87 (d, J = 8.4 Hz, 1H), 6.79 (d, J = 8.4 Hz, 1H), 5.41 (s, 2H), 3.84-3.70 (m, 5H), 3.54 (s, 3H), 2.23- 2.20 (m, 1H), 1.88-1.64 (m, 5H), 1.26-0.98 (m, 4H); ES-LCMS m/z 523 (M+H).
A solution of trans-methyl 4-[(l-{[2-fluoro-3-(trifluoromethyl)phenyl]methyl}-5- methoxy-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (800 mg, 1.53 mmol), KCN (299 mg, 4.59 mmol) in DMSO (10 mL) was stirred at 90 °C overnight. H20 (25 mL) was added to the reaction mixture, and mixture was extracted with DCM (25 mL). The organic layer was concentrated and purified by prepare TLC to give trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methoxy-2,4- dioxo- 1 ,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane- 1 -carboxylate (400 mg, 49.3% yield): 1H NMR (400 MHz, CDC13) δ: 7.51-7.47 (m, 2H), 7.19-7.11 (m, 2H), 6.70 (d, J = 8.4 Hz, 1H), 6.56 (d, J= 8.4 Hz, 1H), 5.39 (s, 2H), 3.94-3.92 (m, 5H), 3.60 (s, 3H), 2.24-2.20 (m, 1H), 1.97-1.77 (m, 5H), 1.39-1.32 (m, 2H), 1.16-1.10 (m,. 2H); ES-LCMS m/z 530 (M+H).
To a stirred solution of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}- 5-methoxy-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l- carboxylate (400 mg, 0.76 mmol) in THF (10 mL) and water (10 mL) was added
LiOH.H20 (192 mg, 4.56 mmol), then the mixture was stirred at room temperature overnight. The mixture was concentrated, acidified by 3 mol/L HC1 to pH 2 and extracted with DCM (30 mL X 2). The combined organic layers were dried over Na2SC"4, filtered and concentrated to give tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5- methoxy-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (195.34 mg, 50.1% yield). 1H NMR (400 MHz, CD3OD + DMSO-d6) δ 7.88 (d, J = 8.0 Hz, 1H), 7.81-7.77 (m, 1H), 7.59 (t, J= 8.4 Hz, 1H), 7.49 (d, J= 8.0 Hz, 1H), 6.95 (d, J= 8.4 Hz, 1H), 6.74 (d, J= 8.0 Hz, 1H), 5.67 (s, 2H), 3.97-3.95 (m, 5H), 2.27-2.20 (m, 1H), 2.06-2.03 (m, 2H), 1.98-1.79 (m, 3H), 1.43-1.30 (m, 2H), 1.19-1.12 (m, 2H); ES- LCMS m/z 516 (M+H).
Example 7: 4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin- -yl)methyl]- 1 -methylcyclohexane- 1 -carboxylic acid
Figure imgf000059_0001
A mixture of methyl 4-(aminomethyl)cyclohexane-l-carboxylate (5 g, 24.15 mmol) and potassium carbonate (6.7 g, 48.3 mmo) in DMF (30 mL) was stirred at room temperature for half an hour, then benzyl bromide (2.92 mL, 24.63 mmol) was added dropwise and the reaction was stirred overnight, then solvent was removed to give the desired compound methyl 4-((benzylamino)methyl)methylcyclohexane-l-carboxylate (6.3 g, 100% yield): 1H NMR (400 MHz, CD3OD) δ 7.48-7.44 (m, 5H), 4.35-4.20 (s, 2H), 3.67 (s, 3H), 2.89-2.80 (m, 3H), 2.55-2.51 (m, 1H), 2.25-2.18 (m, 2H), 1.68-1.60 (m, 2H), 1.18-1.05 (m, 4H); ES- LCMS m/z 262.2 (M+H).
A mixture of 4-((benzylamino)methyl)methylcyclohexane-l-carboxylate (6.3 g, 24.15 mmol) and (Boc)20 (5.37 g, 24.63 mmol) in DCM (200 mL) was stirred at room temperature for overnight, the solvent then was removed to give the residue which was purified by a flash column to give the product methyl 4-((benzyl(tert- butoxycarbonyl)amino)methyl)cyclohexanecarboxylate as a yellow oil (7.7 g, 98% yield): 1H NMR (400 MHz, CD3OD) δ 7.33-7.30 (m, 2H), 7.24-7.20 (m, 3H), 4.46-4.41 (m, 2H), 3.65 (s, 3H), 3.08-2.98 (m, 2H), 2.25-2.18 (m, 1H), 2.01-1.96 (m, 2H), 1.77-1.70 (m, 2H), 1.48-1.44 (m, 9H), 1.00-0.96 (m, 2H); ES-LCMS m/z 306.2 (M+H-56).
To a stirred solution of diisopropylamine (0.7 mL, 12.51 mmol) in THF (30 mL) was added n-BuLi (2.1 mL, 12.51 mmol) at -78 °C under nitrogen. After stirred for 1 hr, a solution of methyl 4-((benzyl(tert-butoxycarbonyl)amino)methyl)cyclohexanecarboxylate (1.5 g, 4.17 mmol) in THF (15 ml) was added dropwise to the mixture and the reaction was stirred at -78 °C for another hour. Mel (0.3 mL, 6.26 mmol) was then added at -78 °C, and the mixture was stirred for overnight at room temperature. The mixture was quenched with water (30 mL), and the solvent was removed to give the residue, which was purified by a flash column to give methyl 4-({benzyl[(tert- butoxy)carbonyl] amino} methyl)- 1-methylcyclohexane-l-carboxylate (2.1 g, 70% yield): 1H NMR (400 MHz, CD3OD) δ 7.34-7.21 (m, 5H), 4.47-4.42 (m, 1H), 3.67 (d, J = 8.4 Hz, 3H), 3.04-2.97 (m, 2H), 2.24-2.17 (m, 2H), 1.60-1.55 (m, 4H), 1.50-1.460 (m, 2H), 1.43 (m, 9H), 1.15-1.12 (m, 3H); ES-LCMS m/z 276.2 (M+H-100).
A solution of methyl 4-({benzyl[(tert-butoxy)carbonyl]amino}methyl)-l- methylcyclohexane-l-carboxylate (2.8 g, 7.76 mmol) in HCl/MeOH (4 N, 20 mL) was stirred at r.t. for 2 hr, then solvent was removed in vacuo to give methyl 4- [(benzylamino)methyl]-l-methylcyclohexane-l-carboxylate (2.0 g, 99% yield): 1H NMR (400 MHz, CD3OD) δ 7.48-7.44 (m, 5H), 4.35-4.20 (s, 2H), 3.67 (s, 3H), 2.89-2.80 (m, 3H), 2.55-2.50 (m, 1H), 2.25-2.18 (m, 2H), 1.68-1.60 (m, 2H), 1.18-1.05 (m, 7H); ES- LCMS m/z 216.2 (M+H).
A mixture of methyl 4-[(benzylamino)methyl]-l-methylcyclohexane-l-carboxylate (3 g, 10.9 mmol) and Pd(OH)2/C (0.5 g) in MeOH (150 mL) was heated to 50 °C under hydrogen (50 psi) overnight, then the mixture was filtered and the filtrate was concentrated to give methyl 4-(aminomethyl)-l-methylcyclohexane-l-carboxylate as a colorless oil (2 g, 98% yield):1H NMR (400 MHz, CD3OD) δ 3.67 (s, 3H), 2.74 (d, 3H), 2.22-2.20 (m, 2H), 1.73-1.60 (m, 4H), 1.25-1.22 (m, 2H), 1.28 (s, 3H), 1.18-1.09 (m, 2H).
To a 200 mL pear-shaped flask containing methyl 4-(aminomethyl)-l-methylcyclohexane- 1-carboxylate (2 g, 10.9 mmol) was added 5-methyl-2,4-dihydro-lH-3,l-benzoxazine-2,4- dione (2 g, 20 mmol), ethanol (60 mL) and water (60 mL). The reaction mixture was allowed to stir at room temperature overnight. LCMS showed presence of desired target. The mixture was concentrated to give the residue, which was purified by a flash column to give methyl 4-{[(2-amino-6-methylphenyl)formamido]methyl}-l-methylcyclohexane-l- carboxylate (1.5 g, 43.2 % yield): 1H NMR (400 MHz, CDC13) δ 6.97-6.93 (m, 1H), 6.50- 6.44 (m, 2H), 5.74 (br, 1H), 4.01 (s, 2H), 3.58 (s, 3H), 3.23-3.20 (m, 2H), 2.24 (s, 3H), 2.18-2.15 (m, 2H), 1.66-1.62 (m, 2H), 1.49-1.44 (m, 1H), 1.15-1.01 (m, 6H); ES-LCMS m/z 319.2 (M+H).
A mixture of methyl 4-{[(2-amino-6-methylphenyl)formamido]methyl}-l- methylcyclohexane-l-carboxylate (0.5 g, 1.57 mmol) and triphosgene (0.23 g, 0.78 mmol) in THF (30 mL) was stirred at r.t overnight. The mixture was then filtered, and the filtrate was washed with water (300 mL) and extracted with DCM (20 mL X 2). The combined organic layers were dried over Na2S04, filtered and concentrated to give methyl 1-methyl- 4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l- carboxylate (0.3 g, 56% yield): 1H NMR (400 MHz, CDC13) δ 7.45-7.41 (m, 1H), 7.00- 6.97 (m, 1H), 6.91-6.89 (m, 1H), 3.89 (d, J= 7.2 Hz, 2H), 3.69 (s, 3H), 2.78 (s, 3H), 2.23- 2.20 (m, 2H), 1.83-1.80 (m, 1H), 1.67-1.65 (m, 2H), 1.21-1.12 (m, 7H); ES-LCMS m/z 345.1 (M+H).
A mixture of methyl l-methyl-4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (344 mg, 1 mmol), l-bromomethyl-2-fluoro-3- trifluoromethyl-benzene (283 mg, 1.1 mmol) and potassium carbonate (276 mg, 2 mmol) in DMF (40 mL) was stirred at r.t. for 16 hr. The mixture was then concentrated in vacuo to give the residue, which was washed with water (200 mL) and extracted with DCM (30 mL X 2). The combined organic layers were dried over Na2S04, filtered and concentrated to give the residue which was purified by a flash column to give methyl 4-[(l-{[2-fluoro- 3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]-l-methylcyclohexane-l-carboxylate (220 mg, 73% yield): 1H NMR (400 MHz, CDCls) δ 7.48-7.45 (m, 1H), 7.35-7.31 (m, 1H), 7.19-7.14 (m, 1H), 7.10-7.06 (m, 1H), 6.98-6.96 (m, 1H), 6.81-6.79 (m, 1H), 5.38 (s, 2H), 3.90 (d, J = 7.2 Hz, 2H), 3.62 (s, 3H), 2.75 (s, 3H), 2.17-2.14 (m, 2H), 1.85-1.80 (m, 1H), 1.78-1.77 (m, 3H), 1.43-1.39 (m, 2H), 1.19-1.15 (m, 5H); ES-LCMS m/z 521.3 (M+H).
To a solution of methyl 4-[(l-{[2-fluoro-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4- dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] - 1 -methylcyclohexane- 1 -carboxylate (300 mg, 0.58 mmol) in MeOH (30 mL) and water (10 mL) was added sodium hydroxide (0.48 g, 12.0 mmol), and the mixture was stirred at 50 °C for 16 h. The mixture was then diluted with cold water (100 mL), acidified and extracted with DCM (20 mL X 3), the combined organic layers were dried over Na2S04, filtered and concentrated to afford 4-[(l-{[2- fluoro-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l, 2,3,4- tetrahydroquinazolin-3-yl)methyl]-l -methylcyclohexane- 1-carboxylic acid as a yellow solid (220 mg, 77%): 1H NMR (400 MHz, CDC13) δ 7.53 (d, J = 7.2 Hz, 1H), 7.43-7.39 (m, 1H), 7.23-7.22 (m, 1H), 7.17-7.14 (m, 1H), 7.05-7.03 (m, 1H), 6.89-6.87 (m, 1H), 5.46 (s, 2H), 3.98 (d, J= 7.2 Hz, 2H), 2.82 (s, 3H), 2.25-2.22 (m, 2H), 1.85-1.81 (m, 1H), 1.66- 1.62 (m, 2H), 1.26-1.14 (m, 7H); ES-LCMS m/z 507.2 (M+H).
To a solution of 4-[(l-{[2-fluoro-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- l,2,3,4-tetrahydroquinazolin-3-yl)methyl]-l-methylcyclohexane-l-carboxylic acid (50 mg, 0.1 mmol) in DMF (25 mL) was added potassium cyanide (33 mg, 0.5 mmol), then the reaction was stirred at 100 °C for 16 hours. Then, the mixture was diluted with cold water (100 mL) and extracted with DCM (20 mL X 3), the combined organic layers were dried over Na2S04, filtered and concentrated to afford the crude product, which was purified by prep HPLC to give 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]-l-methylcyclohexane-l-carboxylic acid as a light yellow solid (11.92 mg, 22% yield): 1H NMR (400 MHz, CDC13) δ 7.72 (d, J = 7.2 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.28-7.27 (m, 1H), 7.05 (d, J = 7.6 Hz, 1H), 6.72 (d, J = 8.4 Hz, 1H), 5.64 (s, 2H), 3.99 (d, J = 7.2 Hz, 2H), 2.82 (s, 3H), 2.24-2.20 (m, 2H), 1.85-1.80 (m, 1H), 1.66-1.62 (m, 2H), 1.26-1.14 (m, 7H); ES- LCMS m/z 536.1 (M+Na).
Example 8: trans-4- [( 1 - { [2-Cyano-3 -(trifluoromethyl)phenyl]methyl} -5 -methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000063_0001
To a solution of trans-methyl 4-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (1.0 g, 3.30 mmol) and potassium carbonate (0.91 g, 6.6 mmol) in CH3CN (10 mL) was added l-bromomethyl-2-cyano-3- trifluoromethylbenzene (0.98 g, 3.80 mmol), and the mixture was stirred at 60-80 °C overnight. Then the mixture was cooled to room temperature and filtered. The filtrate was concentrated and purified by preparative HPLC to give trans-methyl 4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (1.0 g, 65 % yield) as a yellow solid: 1H NMR (400 MHz, CD3OD) δ 7.83 (d, J = 7.6 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.49-7.41 (m, 2H), 7.10 (d, J = 7.6 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 5.66 (s, 2H), 3.97-3.94 (m, 2H), 3.62 (s, 3H), 2.81 (s, 3H), 2.30-2.24 (m, 1H), 2.00-1.79 (m, 5H), 1.38-1.35 (m, 2H), 1.19-1.10 (m, 2H); ES-LCMS m/z 514 (M+H).
To a suspension of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5- methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (0.6 g, 1.17 mmol) in tetrahydrofuran (10 mL) and water (10 mL) was added LiOH.H20 (0.19 g, 4.68 mmol). The mixture was stirred at room temperature for 4 hours. Then con. HCl was added to the mixture until pH~2 was obtained. The mixture was evaporated until THF solvent was removed to give an aqueous suspension mixture, which was filtered, washed with water (5 mL), dried in vacuum to give tra/?5-4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (yield 300 mg, 51% yield): 1H NMR (400 MHz, CDCls) δ 7.73 (d, J= 8.0 Hz, 1H), 7.62 (d, J= 8.0 Hz, 1H), 7.42 (d, J= 8.0 Hz, 1H), 7.29- 7.27 (m, 1H), 7.07 (d, J = 7.6 Hz, 1H), 6.74 (d, J = 8.4 Hz, 1H), 5.65 (s, 2H), 4.01 (d, J = 7.2 Hz, 2H), 2.84 (s, 3H), 2.35-2.27 (m, 1H), 2.07-2.04 (m, 2H), 1.96-1.82 (m, 3H), 1.48- 1.44 (m, 2H), 1.19-1.15 (m, 2H); ES-LCMS m/z 500.1 (M+H).
Example 9: tra/?5-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-6-methoxy-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000064_0001
The mixture of 5-methoxy-2-nitrobenzoic acid (5 g, 25.4 mmol) and Pd/C (0.5 g, 4.70 mmol) in Methanol (100 mL), was stirred at 50 °C for 16 hours. The mixture was filtered, and the filtrate was concentrated to give 2-amino-5-methoxybenzoic acid (4.1 g, 23.30 mmol, 92 % yield) as a yellow solid: 1H NMR (400 MHz, CD3OD) δ 7.36 (d, J = 2.8 Hz, 1H), 6.88 (dd, J = 8.8, 2.8 Hz, 1H), 6.71 (d, J = 8.8 Hz, 1H), 3.71 (s, 3H); ES-LCMS m/z 168 (M+H).
The mixture of trans-methyl 4-(aminomethyl)cyclohexanecarboxylate (2.049 g, 11.96 mmol), 2-amino-5-methoxybenzoic acid (2 g, 11.96 mmol), HOBT (1.832 g, 11.96 mmol), EDC (2.294 g, 11.96 mmol) and DIEA (2.090 mL, 11.96 mmol) in DCM (50 mL), was stirred at 15 °C for 16 hours. The mixture was washed with water, and the organic phase was dried over anhydrous Na2S04, filtered and concentrated to give trans-methyl 4-
((2-amino-5-methoxybenzamido)methyl)cyclohexanecarboxylate (2.5 g, 7.41 mmol, 62.0 % yield) as a yellow solid: 1H NMR (400 MHz, CD3OD) δ 7.00 (d, J = 2.8 Hz, 1H), 6.86 (dd, J= 8.8 Hz, 2.8 Hz, 1H), 6.73 (dd, J = 9.2 Hz, 0.4 Hz, 1H), 3.73 (s, 3H), 3.63 (s, 3H), 3.17 (d, J = 6.8 Hz, 2H), 1.20-1.96 (m, 2H), 1.90-1.86 (m, 2H), 1.42-1.39 (m, 2H), 1.11- 1.02 (m, 4H); ES-LCMS m/z 321 (M+H).
A mixture of trans-methyl 4-((2-amino-5-methoxybenzamido)methyl)cyclohexane carboxylate (2.54 g, 7.93 mmol) and Et3N (2.210 mL, 15.86 mmol) in THF (50 mL) was added triphosgene (1.059 g, 3.57 mmol) slowly. The mixture was stirred at 15 °C for 16 hours. The solvent was concentrated, the residue was washed with water and MeOH, and the solid was dried to give trans -methyl 4-((6-methoxy-2,4-dioxo-l,2-dihydroquinazolin- 3(4H)-yl)methyl)cyclohexanecarboxylate (1.2 g, 3.12 mmol, 39.3 % yield) as a yellow solid: 1H NMR (400 MHz, CD3OD) δ 7.51 (d, J = 2.8 Hz, 1H), 7.29 (dd, J = 8.8 Hz, 2.8 Hz, 1H), 7.15-7.13 (m, 1H), 3.93-3.91 (m, 3H), 3.66-3.62 (m, 5H), 2.39-2.21 (m, 2H), 2.01-1.76 (m, 5H), 1.21-1.02 (m, 4H); ES-LCMS m/z 347 (M+H).
To a mixture of 2-(bromomethyl)-6-(trifluoromethyl)benzonitrile (100 mg, 0.379 mmol) and trans-methyl 4-((6-methoxy-2,4-dioxo-l ,2-dihydroquinazolin-3(4H)- yl)methyl)cyclohexane carboxylate (0.181 mL, 0.379 mmol) in DMF (5 mL), was added K2C03 (157 mg, 1.136 mmol). The mixture was stirred at 10 °C for 16 hours. The mixture was filtered, and the filtrate was concentrated to give trans -methyl 4-((l-(2- cyano-3-(trifluoromethyl)benzyl)-6-methoxy-2,4-dioxo-l,2-dihydroquinazolin-3(4H)- yl)methyl)cyclohexanecarboxylate (190 mg, 0.305 mmol, 81 % yield) as a yellow solid: 1H NMR (400 MHz, CDC13) δ 7.67 (d, J= 8.0 Hz, 1H), 7.64 (d, J= 3.2 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.22-7.71 (m, 1H), 7.12 (dd, J= 9.2, 2.8 Hz, 1H), 6.78 (d, J = 5.2 Hz, 1H), 3.97 (d, J = 7.2 Hz, 2H), 3.81 (s, 3H), 3.60 (s, 3H), 2.89 (s, 2H), 2.20-2.17 (m, 1H), 1.95- 1.92 (m, 2H), 1.85-1.82 (m, 1H), 1.76-1.73 (m, 2H), 1.36-1.35 (m, 2H), 1.20-1.01 (m, 2H); ES-LCMS m/z 530 (M+H).
To a mixture of trans -methyl 4-((l-(2-cyano-3-(trifluoromethyl)benzyl)-6-methoxy-2,4- dioxo-l,2-dihydroquinazolin-3(4H)-yl)methyl)cyclohexanecarboxylate (0.262 mL, 0.359 mmol) in THF (30 mL) and water (10 mL), was added LiOH (43.0 mg, 1.794 mmol) portionwise. The mixture was stirred at 10 °C for 4 hours. 1N HC1 aqueous solution (50 mL) was added. The mixture was extracted with DCM (3 X 50 mL), dried over Na2S04, filtered and concentrated. The resulting residue was purified by prep HPLC to give trans- 4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -6-methoxy-2,4-dioxo- 1 ,2,3 ,4- tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (31.95 mg, 0.060 mmol, 16.60 % yield) as a white solid: 1H NMR (400 MHz, CD3OD) δ 7.90 (d, J = 7.6 Hz, 1H), 7.81 (t, J = 7.8 Hz, 1H), 7.72 (d, J = 3.2 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.33 (dd, J = 9.2 Hz, 3.2 Hz, 1H), 7.20 (d, J = 9.2 Hz, 1H), 5.70 (s, 2H), 4.01 (d, J = 6.8 Hz, 2H), 3.91 (s, 3H), 2.29-2.23 (m, 1H), 2.03-2.01 (m, 2H), 1.91-1.88 (m, 3H), 1.45-1.29 (m, 2H), 1.22- 1.11 (m, 2H); ES-LCMS m/z 516 (M+H).
Example 10: tra/?5-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-6-methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000066_0001
To a suspension of 2-amino-5-methylbenzoic acid (2.116 g, 14 mmol), trans-methyl 4- (aminomethyl)cyclohexanecarboxylate, hydrochloride (4.36 g, 21.00 mmol) and DIEA (7.34 mL, 42.0 mmol) in dichloromethane (50 mL) was added HOBt (2.144 g, 14.00 mmol) and EDC (3.22 g, 16.80 mmol). The mixture was stirred at 25 °C for 16 hr. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (20 mL). The organic layers were combined, dried over anhydrous Na2S04 and filtered. The filtrate was concentrated to give trans-methyl 4-{[(2-amino-5- methylphenyl)formamido]methyl}cyclohexane-l-carboxylate (4.0 g, 94 % yield) as a yellow solid: 1H NMR (400 MHz, CDC13) δ 7.25 (s, 1H), 7.08-6.99 (m, 1H), 6.59 (d, J = 8.4 Hz, 1H), 6.15 (br, 1H), 3.64 (s, 3H), 3.25 (t, J = 6.4 Hz, 2H), 2.28-2.20 (m, 4H), 2.02 1.98 (m, 2H), 1.90-1.86 (m, 2H), 1.62-1.51 (m, 1H), 1.48-1.37 (m, 2H), 1.08-0.98 (m 2H); ES-LCMS m/z 305.3 (M+H). To a suspension of trans-methyl 4-{[(2-amino-5- methylphenyl)formamido]methyl}cyclohexane-l-carboxylate and Et3N (2.75 mL, 19.71 mmol) in tetrahydrofuran (80 mL) was added triphosgene (1.560 g, 5.26 mmol). The mixture was stirred at 25 °C for 16 hr and then quenched with water (50 mL). The mixture was concentrated under vacuum to remove THF and extracted with dichloromethane (50 mL X 3). The organic layer was combined, dried over anhydrous Na2SC"4 and filtered. The filtrate was concentrated to give trans-methyl 4-[(6-methyl- 2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (3.8 g, 88 % yield) as a yellow solid: 1H NMR (400 MHz, CDC13) δ 7.89 (s, 1H), 7.40 (dd, J = 8.4, 1.6 Hz, 1H), 7.01 (t, J = 8.4 Hz, 1H), 3.95 (d, J= 7.2 Hz, 2H), 3.63 (s, 3H), 2.38 (s, 3H), 2.29-2.22 (m, 1H), 1.99-1.78 (m, 5H), 1.42-1.32 (m, 2H), 1.19-1.09 (m, 2H); ES- LCMS m/z 331.2 (M+H).
To a suspension of trans-methyl 4-[(6-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (120 mg, 0.363 mmol) and K2C03 (100 mg, 0.726 mmol) in anhydrous N,N-dimethylformamide (5 mL) was added 2-(bromomethyl)-6- (trifluoromethyl)benzonitrile (101 mg, 0.381 mmol), and the mixture was stirred at 60 °C for 16 hr. The suspension was cooled to room temperature and filtered. The filtrate was concentrated and purified by preparative TLC to give trans-methyl 4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-6-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (160 mg, 86 % yield) as a yellow solid: 1H NMR (400 MHz, DMSO-d6) δ 7.92-7.88 (m, 2H), 7.77 (t, J = 8.0 Hz, 1H), 7.49-7.45 (m, 2H), 7.22 (d, J = 8.8 Hz, 1H), 5.55 (s, 2H), 3.80 (d, J = 6.8 Hz, 2H), 3.53 (s, 3H), 2.86 (s, 3H), 2.25-2.18 (m, 1H), 1.85-1.83 (m, 2H), 1.70-1.64 (m, 3H), 1.24-1.05 (m, 4H); ES-LCMS m/z 514.3 (M+H).
To a suspension of trans-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6- methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (160 mg, 0.243 mmol) in tetrahydrofuran (4 mL) and water (4 mL) was added lithium hydroxide.H20 (51.1 mg, 1.217 mmol). The mixture was stirred at 25 °C for 4 hr. The mixture was quenched with HCl solution (8 mL) and extracted with dichloromethane (15 mL X 2). The organic layers were combined, concentrated and purified by preparative HPLC to give tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6-methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid (58 mg, 47.7 % yield) as a off-white solid: 1H NMR (400 MHz, CDC13) δ 8.04 (s, 1H), 7.84-7.73 (m, 2H), 7.52-7.49 (m, 1H), 7.40 (d, J= 7.6 Hz, 1H), 7.03 (d, J= 8.0 Hz, 1H), 5.68 (s, 2H), 4.00 (d, J= 7.2 Hz, 2H), 2.41 (s, 3H), 2.27-2.21 (m, 1H), 2.03-2.00 (m, 2H), 1.90-1.80 (m, 3H), 1.44-1.33 (m, 2H), 1.21-1.12 (m, 2H); ES-LCMS m/z 500.3 (M+H).
Example 11 : trans-4-[(6-Chloro- 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylic acid
Figure imgf000068_0001
To a solution of Nl-((ethylimino)methylene)-N3,N3-dimethylpropane-l,3-diamine hydrochloride (3.36 g, 17.52 mmol), 2-amino-5-chlorobenzoic acid (3.01 g, 17.52 mmol), (lH-benzo[d][l,2,3]triazol-l-yl)holmium (4.96 g, 17.52 mmol) in Dichloromethane (DCM) (200 mL) was added N-ethyl-N-isopropylpropan-2-amine (6.79 g, 52.6 mmol) at 20 °C. The mixture was stirred at 20 °C for 12 hr. Then trans-methyl 4- (aminomethyl)cyclohexanecarboxylate (3 g, 17.52 mmol) was added, and the mixture was stirred at 20 °C. After LCMS showed that the reaction was complete, the mixture was dissolved in DCM (300 mL) and washed with water (200 mL X 3). The organic phase was dried over Na2S04, filtered and concentrated under vacuum to give trans-methyl 4-((2- amino-5-chlorobenzamido)methyl)cyclohexanecarboxylate (4 g, 12.32 mmol, 70.3 % yield) as a solid. 1H NMR 1H NMR (400 MHz, CDC13) δ 7.26-7.25 (d, J = 2.8 Hz, 1H), 7.16-7.13 (dd, J= 8.8 Hz, 2.4 Hz, 1H), 6.63-6.61 (d, J = 8.8 Hz, 1H), 5.48 (s, 2H), 3.66 (s, 3H), 3.28-3.24 (t, J = 4.8 Hz, 1H), 2.30-2.28 (t, J = 3.6 Hz, 1H), 2.05-2.01 (m, 2H), 1.90- 1.87 (m, 2H), 1.60-1.58 (m, 2H), 1.11-0.92 (m, 2H), ES-LCMS m/z: 325 (M+H).
To a mixture of trans-methyl 4-((2-amino-5-chlorobenzamido)methyl)- cyclohexanecarboxylate (4 g, 12.32 mmol), triethylamine (2.492 g, 24.63 mmol) in tetrahydrofuran (THF) (300 mL) was added triphosgene (1.21 g, 4.12 mmol) at 20 °C. Then, the mixture was stirred at 20 °C for 20 hr. LCMS showed that the reaction was complete, and the mixture was dissolved in DCM (300 mL) and washed with water (200 mL X 3). The organic phase was dried over Na2S04, filtered and concentrated under vacuum to give trans-methyl 4-[(6-chloro-2-oxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (3 g, 8.91 μιηοΐ, 72 % yield) as a solid: 1H NMR (400 MHz, DMSO-d6) δ 11.67 (s, 1H), 7.86-7.85 (d, J = 2.4 Hz, 1H), 7.73-7.70 (dd, J = 8.8 Hz, 2.4 Hz, 1H), 7.27-7.25 (d, J = 8.8 Hz, 1H), 3.77-3.75 (d, J = 7.2 Hz, 1H), 3.37 (s, 3H), 2.52-2.51 (t, J = 1.2 Hz, 1H), 1.90-1.87 (m, 2H), 1.71-1.68 (m, 3H), 1.26-1.23 (m, 2H), 1.07-1.04 (m, 2H), ES-LCMS m/z: 337 (M+H).
To a mixture of trans-methyl 4-[(6-chloro-2-oxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (100 mg, 0.285 mmol) in N,N-Dimethylformamide (DMF) (100 mL) was added 2-(bromomethyl)-6-(trifluoromethyl)benzonitrile (90 mg, 0.342 mmol), K2C03 (79 mg, 0.570 mmol) at 60 °C. Then, the mixture was stirred at 60 °C for 8 hr. LCMS showed that the reaction was complete, and the mixture was dissolved in DCM (300 mL) and washed with water (200 mL X 3). The organic phase was dried over Na2S04, filtered and concentrated under vacuum to give trans-methyl 4-[(6-chloro-l- {[2-cyano-3-(trifluoromethyl)phenyl]methyl} -2-oxo-l,2, 3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (120 mg, 79 % yield) as a solid: 1H NMR (400 MHz, CDC13) δ 8.24 (d, J = 2.4 Hz, 1H), 7.77-7.75 (m, 1H), 7.67-7.63 (m, 1H), 7.56-7.54 (d, J = 7.6 Hz, 1H), 7.28-7.26 (m, 1H), 6.98-6.86 (m, 1H), 5.79 (s, 2H), 4.03-3.99 (t, J = 8.8 Hz, 1H), 3.66 (s, 3H), 2.30-2.24 (t, J = 12.0 Hz, 1H), 2.02-1.99 (m, 2H), 1.77-1.67 (m, 2H), 1.57 (s, 2H), 1.44-1.41 (m, 3H), 1.39-1.31 (m, 2H), 0.89-0.86 (m, 2H), ES-LCMS m/z: 535 (M+H).
To a mixture of trans -methyl 4-[(6-chloro-l-{[2-cyano-3-
(trifluoromethyl)phenyl]methyl}-2-oxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (50 mg, 0.094 mmol) in tetrahydrofuran (THF) (50 mL) and water (30 mL) was added LiOH (6.73 mg, 0.281 mmol) at room temperature. Then, the mixture was stirred at room temperature for 10 hr. LCMS showed that the reaction was completed. The mixture was treated with DCM (300 mL), and the organic layer was washed with water (200 mL X 3). The organic phase was dried over Na2S04 and purified by prep HPLC to give tra/?5-4-[(6-chloro-l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-2-oxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylic acid (17 mg, 0.033 mmol, 34.8 % yield) as a solid: 1H NMR (400 MHz, CD3OD) δ 8.14 (d, J= 2.4 Hz, 1H), 7.85-7.83 (m, 1H), 7.76-7.72 (m, 1H), 7.64 (d, J= 2.4 Hz, 1H), 7.62 (d, J = 2.8 Hz, 1H), 7.19-7.18 (d, J = 8.8 Hz, 1H), 6.64 (s, 2H), 3.95 (d, J = 7.2 Hz, 2H), 2.21-2.10 (t, J = 8.0 Hz, 1H), 1.99-1.96 (m, 2H), 1.84- 1.77 (m, 3H), 1.38-1.32 (m, 2H), 1.29-1.24 (m, 2H), ES-LCMS m/z: 521 (M+H).
Examples 12-13 (Table 1) were prepared using procedures analogous to those described in Example 8, starting from 4-(5-methyl-2,4-dioxo-l,4-dihydro-2H-quinazolin-3-ylmethyl) cyclohexanecarboxylic acid methyl ester (Intermediate 5) and the appropriately substituted benzyl bromide. Table 1 :
Figure imgf000070_0001
Figure imgf000070_0002
1H NMR (400 MHz, CD3OD) δ 7.38 (d, J =
8.4 Hz, 1H), 7.35-7.30 (m, 1H), 7.24-7.20
(m, 1H), 7.03 (d, J= 7.6 Hz, 1H), 6.86 (d, J
= 7.6 Hz, 1H), 6.77 (d, J= 8.4 Hz, 1H), ES-LCMS
13
5.56 (s, 2H), 4.01 (d, J= 8.4 Hz, 2H), 2.82 m/z 446.1 (s, 3H), 2.61 (s, 3H), 2.30-2.26 (m, 1H),
2.06-2.03 (m, 2H), 1.94-1.88 (m, 3H), 1.46- 5
1.38 (m, 2H), 1.20-1.15 (m, 2H).
Example 14: c 5-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid
Figure imgf000071_0001
Step 1 : cis Methyl 4-(hydroxymethyl)cyclohexane-l-carboxylate
Figure imgf000071_0002
To a mixture of czs-4-(hydroxymethyl)cyclohexanecarboxylic acid (2 g, 12.64 mmol) in methanol (50 mL), was added SOCl2 (1.846 mL, 25.3 mmol) dropwise. The mixture was stirred at 25 °C for 16 hr. Then the mixture was concentrated to give czs-methyl 4-
(hydroxymethyl)cyclohexane-l-carboxylate (2.2 g, 12.77 mmol, 101% yield): 1H NMR (400 MHz, CDC13) δ 4.32 (s, 3H), 3.25-3.21 (m, 2H), 2.41-2.39 (m, 1H), 1.82-1.79 (m, 2H), 1.59-1.39 (m, 5H), 1.17-1.11 (m, 2H); ES-LCMS m/z 173 (M+H). - Methyl 4-[(methanesulfonyloxy)methyl]cyclohexane- 1 -carboxylate
MsO.
Figure imgf000072_0001
A mixture of cz's-methyl 4-(hydroxymethyl)cyclohexane-l -carboxylate, MsCl (0.339 mL, 4.35 mmol) and Et3N (1.214 mL, 8.71 mmol) in DCM (10 mL), was stirred at 25 °C for 0.5 hr. The mixture was washed with H20 (2 x 10 mL), dried over Na2S04, filtered, and the filtrate was concentrated to give cz's-methyl 4-
[(methanesulfonyloxy)methyl]cyclohexane-l -carboxylate (550 mg, 1.868 mmol, 64.3% yield) as yellow oil: 1H NMR (400 MHz, CDC13) δ 4.05 (d, J= 6.8 Hz, 2H), 3.68 (s, 3H), 2.99 (s, 3H), 2.61-2.58 (m, 1H), 2.06-2.01 (m, 2H), 1.88-1.83 (m, 1H), 1.69-1.57 (m, 5H), 142-1.26 (m, 3H); ES-LCMS m/z 251 (M+H).
Step 3 : cz's-Methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4- dioxo-1, 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylate
Figure imgf000072_0002
A mixture of 2-[(5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-l-yl)methyl]-6-
(trifluoromethyl)benzonitrile (500 mg, 1.392 mmol), cz's-methyl 4-
[(methanesulfonyloxy)methyl]cyclohexane-l -carboxylate (522 mg, 2.087 mmol) and
K2C03 (577 mg, 4.17 mmol) in DMF (10 mL), was stirred at 60 °C for 16 hr. The mixture was filtered, and the filtrate was concentrated to give cz's-methyl 4-[(l-{[2-cyano-3-
(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l-carboxylate (260 mg, 0.405 mmol, 29.1% yield) as a yellow solid: 1H NMR (400 MHz, CDC13) δ 7.71 (d, J= 8.0 Hz, 1H), 7.63-7.59 (m, 1H), 7.41- 7.37 (m, 1H), 7.27 (s, 1H), 7.05 (d, J= 7.6 Hz, 1H), 6.72 (d, J= 8.8 Hz, 1H), 5.63 (s, 2H), 4.03 (d, J= 7.2 Hz, 2H), 3.67 (s, 3H), 2.94 (s, 3H), 2.57-2.53 (m, 1H), 2.08-1.99 (m, 4H), 1.59-1.55 (m, 3H), 1.41-1.34 (m, 2H); ES-LCMS m/z 514 (M+H).
Step 4: cz5-4-[(l-{[2-Cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid
Figure imgf000073_0001
A mixture of czs-methyl 4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4- dioxo-l,2,3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l-carboxylate (260 mg, 0.506 mmol) and LiOH (60.6 mg, 2.53 mmol) in THF (10 mL) and water (10.00 mL), was stirred at 25 °C for 16 hr. The mixture was adjusted to pH = 2 with 1NHC1 solution and THF was removed by evaporation. The mixture was filtered and the solid was purified by prep-HPLC to give c 5-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4- dioxo-1, 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylic acid (166.95 mg, 0.332 mmol, 65.6% yield) as a white solid: 1H NMR (400 MHz, CDC13) δ 7.73 (d, J =
7.6 Hz, 1H), 7.63 (t, J= 8.0 Hz, 1H), 7.41 (t, J= 8.0 Hz, 1H), 7.29 (d, J= 7.6 Hz, 1H),
7.07 (d, J= 7.6 Hz, 1H), 6.74 (d, J= 8.4 Hz, 1H), 5.65 (s, 2H), 4.06 (d, J= 7.2 Hz, 2H), 2.84 (s, 3H), 2.64-2.62 (m, 1H), 2.16-2.11 (m, 2H), 2.02-2.01 (m, 1H), 1.63-1.57 (m, 4H),
1.47-1.44 (m, 2H); ES-LCMS m/z 500 (M+H). BIOCHEMICAL ASSAYS
Inhibition of human TN S1 [or TN S 2] Fluorescence Polarization (FP) activity in vitro
Test compounds are plated at 0.1 μΐ, diluted in 100% DMSO in low volume 384 well black polypropylene plates (NUNC 267461). A positive control (XAV939) is added to column 18, to a final assay concentration of 20 uM, to define the maximal effect. The ligand solution is prepared as 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM CHAPS, 1 mM DTT, 50 mM NaCl, and 1 nM ligand final concentrations. To this solution, TNKS 1 [or TNKS2] enzyme (5 nM final concentration) is added to prepare the ligand/enzyme solution. The assay is initiated upon the transfer 10 of the enzyme/ligand solution to each well of the compound plates using a Multidrop or similar instrument capable of accurately dispensing 10 μί. Plates are then centrifuged for 1 minute at 500 x g. Assay plates are incubated for 1 to 2 hours at RT and then read on the Analyst GT using the PvhGr 505 filter set (ex at 485 nm, em at 530 nm, 505 dichroic) in the fluorescence polarization mode. Data generated are normalized to a maximal and no effect control. Potency of test molecules are reported as pIC50s (-log(IC50)).
Inhibition of human TN S 1 or TNKS 2 HTRF activity in vitro
Test compounds are plated at 0.1 uL diluted in 100% DMSO in low volume 384 well black polystyrene plates (Greiner 784076). One microliter of a positive control inhibitor (4-({ l-[(2-cyanophenyl)methyl]-2,4-dioxo-l ,2,3,4-tetrahydroquinazolin-3- yl}methyl)-N-(pyridin-4-yl)cyclohexane-l-carboxamide) is added to column 18, to a final assay concentration of 100 nM to define maximal effect. The reaction buffer solution is prepared as 50 mM HEPES (pH 7.5) and 1 mM CHAPS. To this solution, TNKS 1 or TNKS2 enzyme (8 nM final concentration) is added to prepare the enzyme solution. The assay is initiated upon the transfer 5 uL of the 2X enzyme solution to each well of the compound plates using a Multidrop or similar instrument capable of accurately dispensing 5 uL. Plates are then incubated for 30 minutes. A 2X substrate solution is prepared by adding 0.2 mM NAD, 50 nM GST-tev-Axin2 and lOmM MnC12 to the reaction buffer, and 5uL of this solution is added to the enzyme-test compound plate with a multidrop or similar instrument. Plates are then centrifuged for 1 minute at 500 x g, and incubated for 2 hours at RT. Finally, 2X detection mix is prepared, consisting of premade HTRF buffer
(50 mM hepes, pH 7.0, 0.1% BSA, 0.8M potassium fluoride, 20 mM EDTA) with 25 nM anti-GST-d2, 1 nM Eu anti-PAR and 10 mM nicotinamide. 10 uL of 2X detection mix is added to each well and incubated for 1 hour. Assay plates are read on the Viewlux (ex at 337 nm, em at 618 nm, and 671nm, plus or minus 8nm), and data are normalized to a maximal and no effect control. The potency of test molecules are reported as pICsoS (- logflCso)).
Biochemical Data
The compounds of Examples 1-14 of the present invention were tested according to the assays described above and were found to be inhibitors of TANKYRASE with pIC5o >6 in one or both TNKS assays.
Each compound listed below was tested two or more times generally according to the assays described herein, and the average pIC5o values are listed in the table below.
Figure imgf000075_0001
References:
Wynn T. A. Cellular and molecular mechanisms of fibrosis. J. Pathol. 214, 199-210 (2008).
Strieter R. M. & Mehrad B. New mechanisms of pulmonary fibrosis. Chest 136, 1364- 1370 (2009).
Verrecchia F. & Mauviel A. Transforming growth factor-beta and fibrosis. World J.
Gastroenterol. 13, 3056-3062 (2007).
Sonnylal S. et al. Postnatal induction of transforming growth factor beta signaling in fibroblasts of mice recapitulates clinical, histologic, and biochemical features of scleroderma. Arthritis Rheum. 56, 334-344 (2007).
Akhmetshina A, Palumbo K, Dees C, Bergmann C, Venalis P, Zerr P, Horn A, Kireva T, Beyer C, Zwerina J, Schneider H, Sadowski A, Riener MO, Macdougald OA, Distler O, Schett G, Distler JH. Activation of canonical Wnt signalling is required for TGF-β- mediated fibrosis. Nat Commun. 1-12 (2012). Chilosi M. et al. Aberrant Wnt/beta-catenin pathway activation in idiopathic pulmonary fibrosis. Am. J. Pathol. 162, 1495-1502 (2003).
Colwell A. S., Krummel T. M., Longaker M. T. & Lorenz H. P. Wnt-4 expression is increased in fibroblasts after TGF-betal stimulation and during fetal and postnatal wound repair. Plast. Reconstr. Surg. I ll, 2297-2301 (2006). He W. et al. Wnt/beta-catenin signaling promotes renal interstitial fibrosis. J. Am. Soc. Nephrol. 20, 765-776 (2009).
He W. et al. Exogenously administered secreted frizzled related protein 2 (Sfrp2) reduces fibrosis and improves cardiac function in a rat model of myocardial infarction. Proc. Natl. Acad. Sci. USA 107, 21110-21115 (2010). Henderson W. R. Jr. et al. Inhibition of Wnt/beta-catenin/CREB binding protein (CBP) signaling reverses pulmonary fibrosis. Proc. Natl. Acad. Sci. USA 107, 14309-14314 (2010).
Konigshoff M. et al. Functional Wnt signaling is increased in idiopathic pulmonary fibrosis. PLoS One 3, e2142 (2008). Liu L. et al. Wnt pathway in pulmonary fibrosis in the bleomycin mouse model. J.
Environ. Pathol. Toxicol. Oncol. 28, 99-108 (2009).
Surendran K., McCaul S. P. & Simon T. C. A role for Wnt-4 in renal fibrosis. Am. J. Physiol. Renal. Physiol. 282, F431-F441 (2002). Trensz F., Haroun S., Cloutier A., Richter M. V. & Grenier G. A muscle resident cell population promotes fibrosis in hindlimb skeletal muscles of mdx mice through the Wnt canonical pathway. Am. J. Physiol. Cell Physiol. 299, C939-C947 (2010).
Wei J. et al. Canonical Wnt signaling induces skin fibrosis and subcutaneous lipoatrophy: a novel mouse model for scleroderma? Arthritis Rheum. 63, 1707-1717 (2011).
Ulsamer, A., Wei, Y., Kim, K. K,, Tan, K,, Wheeler, S., Xi, Y., Thies, S., and Chapman, H.A. Axin Pathway Activity Regulates in Vivo pY 654~p-eatenm Accumulation and Pulmonary Fibrosis. J. Biol. Chem. 2012 287: 5164-5172. (201 1 )

Claims

Claims:
1. A compound of Formula (I):
Figure imgf000078_0001
(I)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy; R4 is hydrogen or methyl;
provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
2. A compound of Formula (I)(a):
Figure imgf000078_0002
(0(a)
wherein
R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy; R4 is hydrogen or methyl;
provided that at least one of R1, R2 and R3 is not hydrogen;
or a pharmaceutically acceptable salt thereof.
3. The compound of claim 1 re resented by Formula (II):
Figure imgf000079_0001
(Π)
wherein
R1, R2, and R3 are defined as in claim 1 ; or a pharmaceutically acceptable salt thereof.
4. The compound of claim 2 re resented by Formula (II)(a):
Figure imgf000079_0002
(II)(a)
wherein
R1, R2, and R3 are defined as in claim 2; or a pharmaceutically acceptable salt thereof.
5. The compound according to any one of claims 1-4, wherein R1 is hydrogen; R2 and R3 are each independently hydrogen, halo, Ci-C4alkyl or Ci-C4alkoxy; and R4 is hydrogen; provided that at least one of R2 and R3 is not hydrogen; or a pharmaceutically acceptable salt thereof.
6. The compound according to any one of claims 1-4, wherein R2 is hydrogen; R1 and R3 are each independently hydrogen, halo, Ci-C4alkyl or Ci-C4alkoxy; and R4 is hydrogen; provided that at least one of R1 and R3 is not hydrogen; or a pharmaceutically acceptable salt thereof.
7. The compound according to any one of claims 1-4, wherein R3 is hydrogen; R1 and R2 are each independently hydrogen, halo, Ci-C4alkyl, or Ci-C4alkoxy; and R is hydrogen; provided that at least one of R1 and R2 is not hydrogen; or a pharmaceutically acceptable salt thereof.
8. The compound according to any one of claims 1-7, wherein said substituted Ci- C4alkyl is Ci-C4haloalkyl; or a pharmaceutically acceptable salt thereof.
9. The compound of claim 8, wherein said Ci-C4haloalkyl is CF3; or a
pharmaceutically acceptable salt thereof.
10. The compound of claim 1 which is:
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4- tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/?s-4-[(5-chloro- 1 - {[2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-fluoro-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-6-fluoro-5-methyl-2,4- dioxo-1 , 2, 3,4-tetrahydroquinazolin-3-yl)methyl]cyclohexane-l -carboxylic acid;
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -5 ,6-dimethyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methoxy-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
4-[(l- {[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l , 2,3,4- tetrahydroquinazolin-3 -yl)methyl] - 1 -methylcyclohexane- 1 -carboxylic acid;
tra/75-4-[(l-{[2-cyano-3-(trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid;
trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -6-methoxy-2,4-dioxo-
1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl] cyclohexane- 1 -carboxylic acid; trans-4-[( 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -6-methyl-2,4-dioxo- 1 ,2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
tra/?s-4-[(6-chloro- 1 - { [2-cyano-3-(trifluoromethyl)phenyl]methyl} -2,4-dioxo-
I , 2,3 ,4-tetrahydroquinazolin-3 -yl)methyl]cyclohexane- 1 -carboxylic acid;
tra/75-4-({l-[(3-chloro-2-cyanophenyl)methyl]-5-methyl-2,4-dioxo-l, 2,3,4- tetrahydroquinazolin-3-yl}methyl)cyclohexane- 1 -carboxylic acid; or
trans-4-( { 1 -[(2-cyano-3-methylphenyl)methyl]-5-methyl-2,4-dioxo- 1 ,2,3,4- tetrahydroquinazolin-3-yl}methyl)cyclohexane- 1 -carboxylic acid;
or a pharmaceutically acceptable salt thereof.
I I . The compound of claim 2 which is cz5-4-[(l-{[2-cyano-3- (trifluoromethyl)phenyl]methyl}-5-methyl-2,4-dioxo-l,2,3,4-tetrahydroquinazolin-3- yl)methyl]cyclohexane-l -carboxylic acid or a pharmaceutically acceptable salt thereof.
12. A pharmaceutical composition comprising the compound according to any one of claims 1-11 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
13. A method of treating cancer comprising administering an effective amount of the compound according to any one of claims 1-11 or a pharmaceutically acceptable salt thereof to a human in need thereof.
PCT/US2013/042332 2012-05-25 2013-05-23 Quinazolinediones as tankyrase inhibitors WO2013177349A2 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201261651687P 2012-05-25 2012-05-25
US61/651,687 2012-05-25
US201361773300P 2013-03-06 2013-03-06
US61/773,300 2013-03-06

Publications (2)

Publication Number Publication Date
WO2013177349A2 true WO2013177349A2 (en) 2013-11-28
WO2013177349A3 WO2013177349A3 (en) 2014-01-16

Family

ID=49624519

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2013/042332 WO2013177349A2 (en) 2012-05-25 2013-05-23 Quinazolinediones as tankyrase inhibitors

Country Status (1)

Country Link
WO (1) WO2013177349A2 (en)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015150449A2 (en) 2014-04-02 2015-10-08 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
US9193689B2 (en) 2012-03-07 2015-11-24 Institute Of Cancer Research: Royal Cancer Hospital (The) 3-aryl-5-substituted-isoquinolin-1-one compounds and their therapeutic use
US9611223B2 (en) 2013-09-11 2017-04-04 Institute Of Cancer Research: Royal Cancer Hospital (The) 3-aryl-5-substituted-isoquinolin-1-one compounds and their therapeutic use
WO2017055313A1 (en) 2015-10-01 2017-04-06 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
WO2017055316A1 (en) 2015-10-01 2017-04-06 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
WO2018078009A1 (en) 2016-10-29 2018-05-03 Bayer Pharma Aktiengesellschaft Amido-substituted cyclohexane derivatives
WO2018078005A1 (en) 2016-10-29 2018-05-03 Bayer Pharma Aktiengesellschaft Amido-substituted azaspiro derivatives as tankyrase inhibitors
WO2018087126A1 (en) 2016-11-09 2018-05-17 Bayer Pharma Aktiengesellschaft Amido-substituted cyclohexane derivatives as inhibitors of tankyrase
US10722484B2 (en) 2016-03-09 2020-07-28 K-Gen, Inc. Methods of cancer treatment
CN112624911A (en) * 2021-01-15 2021-04-09 阿里生物新材料（常州）有限公司 2-chloro-3-fluoro-4- (trifluoromethyl) benzaldehyde and synthetic method thereof
CN112898221A (en) * 2019-12-03 2021-06-04 浙江京新药业股份有限公司 Preparation method and intermediate of benzoxazine diketone compound C

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5634263B2 (en) * 2007-08-16 2014-12-03 ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング Quinazolinedione chymase inhibitor
AU2009260503B2 (en) * 2008-05-27 2014-03-27 The Board Of Regents Of The University Of Texas System Wnt protein signalling inhibitors
FR2943673B1 (en) * 2009-03-27 2013-03-29 Sanofi Aventis THERAPEUTIC APPLICATIONS OF QUINAZOLINEDIONE DERIVATIVES
US8865704B2 (en) * 2009-10-07 2014-10-21 Siena Biotech S.P.A. Wnt pathway antagonists

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9193689B2 (en) 2012-03-07 2015-11-24 Institute Of Cancer Research: Royal Cancer Hospital (The) 3-aryl-5-substituted-isoquinolin-1-one compounds and their therapeutic use
US9611223B2 (en) 2013-09-11 2017-04-04 Institute Of Cancer Research: Royal Cancer Hospital (The) 3-aryl-5-substituted-isoquinolin-1-one compounds and their therapeutic use
WO2015150449A2 (en) 2014-04-02 2015-10-08 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
US9884063B2 (en) 2014-04-02 2018-02-06 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
WO2017055313A1 (en) 2015-10-01 2017-04-06 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
WO2017055316A1 (en) 2015-10-01 2017-04-06 Bayer Pharma Aktiengesellschaft Amido-substituted azole compounds
US10722484B2 (en) 2016-03-09 2020-07-28 K-Gen, Inc. Methods of cancer treatment
WO2018078005A1 (en) 2016-10-29 2018-05-03 Bayer Pharma Aktiengesellschaft Amido-substituted azaspiro derivatives as tankyrase inhibitors
WO2018078009A1 (en) 2016-10-29 2018-05-03 Bayer Pharma Aktiengesellschaft Amido-substituted cyclohexane derivatives
WO2018087126A1 (en) 2016-11-09 2018-05-17 Bayer Pharma Aktiengesellschaft Amido-substituted cyclohexane derivatives as inhibitors of tankyrase
CN112898221A (en) * 2019-12-03 2021-06-04 浙江京新药业股份有限公司 Preparation method and intermediate of benzoxazine diketone compound C
CN112898221B (en) * 2019-12-03 2023-12-08 浙江京新药业股份有限公司 Preparation method of benzoxazine diketone compound C and intermediate thereof
CN112624911A (en) * 2021-01-15 2021-04-09 阿里生物新材料（常州）有限公司 2-chloro-3-fluoro-4- (trifluoromethyl) benzaldehyde and synthetic method thereof

Also Published As

Publication number Publication date
WO2013177349A3 (en) 2014-01-16

Similar Documents

Publication Publication Date Title
WO2013177349A2 (en) Quinazolinediones as tankyrase inhibitors
US10478426B2 (en) Enhancer of Zeste Homolog 2 inhibitors
EP2970191B1 (en) Pyridine derivatives as rearranged during transfection (ret) kinase inhibitors
EP3191449B1 (en) Compounds as rearranged during transfection (ret) inhibitors
JP6538154B2 (en) Pyridine derivatives as RET (REARRANGED DURING TRANSFECTION) kinase inhibitors
EP2330909B1 (en) Chemical compounds
EP2755962B1 (en) Azaindazoles
US8637509B2 (en) Azaindazoles
AU2015340614B2 (en) Enhancer of zeste homolog 2 inhibitors
EP3019494B1 (en) Enhancer of zeste homolog 2 inhibitors
US9382234B2 (en) Enhancer of Zeste Homolog 2 inhibitors
US20150099762A1 (en) Compounds as rearranged during transfection (ret) inhibitors
US9084794B2 (en) Fatty acid synthase inhibitors
US20150273057A1 (en) Combination
WO2013028445A1 (en) Fatty acid synthase inhibitors
US20130172384A1 (en) Fatty acid synthase inhibitors
EP3114125A1 (en) Enhancer of zeste homolog 2 inhibitors
EP2943484B1 (en) Fatty acid synthase inhibitors

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: 13794700

Country of ref document: EP

Kind code of ref document: A2

122 Ep: pct application non-entry in european phase

Ref document number: 13794700

Country of ref document: EP

Kind code of ref document: A2