Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
  • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
  • C07D495/04—Ortho-condensed systems
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• the present invention is directed to novel 4-amino-thieno[3,2-c]pyridine-7- carboxylic acid amides and their pharmaceutically acceptable salts and esters. These compounds inhibit KDR (kinase insert domain-containing receptor) kinase and/or FGFR (fibroblast growth factor receptor) kinase. These compounds and their pharmaceutically acceptable salts and esters have antiproliferative activity and are useful in the treatment or control of cancer, in particular solid tumors. In addition these compounds have advantageous bioavailability profiles.
• This invention is also directed to pharmaceutical compositions containing such compounds and to methods of treating or controlling cancer, most particularly the treatment or control of breast, lung, colon and prostate tumors.
• Protein kinases are a class of proteins (enzymes) that regulate a variety of cellular functions. This is accomplished by the phosphorylation of specific amino acids on protein substrates resulting in conformational alteration of the substrate protein. The conformational change modulates the activity of the substrate or its ability to interact with other binding partners.
• the enzyme activity of the protein kinase refers to the rate at which the kinase adds phosphate groups to a substrate. It can be measured, for example, by determining the amount of a substrate that is converted to a product as a function of time. Phosphorylation of a substrate occurs at the active-site of a protein kinase.
• Tyrosine kinases are a subset of protein kinases that catalyze the transfer of the terminal phosphate of adenosine triphosphate (ATP) to tyrosine residues on protein substrates. These kinases play an important part in the propagation of growth factor signal transduction that leads to cellular proliferation, differentiation and migration.
• ATP adenosine triphosphate
• FGF basic fibroblast growth factor
• VEGF vascular endothelial growth factor
• KDR kinase insert domain-containing receptor
• inhibitors of the receptors FGFR and/or KDR that interfere with the growth signal transduction, and thus slow down or prevent angiogenesis are useful agents in the prevention and treatment of solid tumors. See Klohs W.E. et. al., Current Opinion in Biotechnology 1999, 10, p.544.
• the present invention relates to novel 4-amino- thieno[3,2-c]pyridine-7-carboxylic acid amides capable of selectively inhibiting the activity of KDR and/or FGFR. These compounds are useful in the treatment or control of cancer, in particular the treatment or control of solid tumors.
• this invention relates to compounds of formula
• R 1 and R 2 are as hereinafter defined.
• the present invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds of formula I, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or excipient.
• the present invention further relates to a method for treating or controlling solid tumors, in particular treatment or control of breast, lung, colon and prostate tumors, most particularly breast or colon tumors, by administering to a human patient in need of such therapy an effective amount of a compound of formula I and/or a pharmaceutically acceptable salt thereof.
• the present invention is further directed to novel intermediate compounds useful in the preparation of compounds of formula I.
• Alkyl denotes a straight-chain or branched saturated aliphatic hydrocarbon having 1 to 10, preferably 1 to 6, and more preferably 1 to 4 carbon atoms. Alkyl groups having 1 to 6 carbon atoms are also referred to herein as "lower alkyl.” Typical lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, t-butyl, 2- butyl, pentyl and hexyl. As used herein the sample designation Ci -4 alkyl means alkyl having from 1 to 4 carbon atoms.
• Aryl means an aromatic carbocyclic radical, for example a 6 -10 membered aromatic or partially aromatic ring system.
• Preferred aryl groups include, but are not limited to, phenyl, naphthyl, tolyl and xylyl.
• Cycloalkyl means a non-aromatic, partially or completely saturated cyclic aliphatic hydrocarbon group containing 3 to 8 atoms.
• Examples of cycloalkyl groups include cyclopropyl, cyclopentyl and cyclohexyl.
• Effective amount or “Therapeutically effective amount” means an amount of at least one compound for formula I, or a pharmaceutically acceptable salt thereof, that significantly inhibits tumor growth.
• Halogen means fluorine, chlorine, bromine or iodine, preferably bromine, chlorine or fluorine.
• Hetero atom means an atom selected from N, O and S, preferably N. [ If the hetero atom is N, it can be present as -NH- or -N-lower alkyl-. If the hetero atom is S, it can be present as S, SO or SO 2 .
• Heteroaryl means an aromatic heterocyclic ring system containing up to two rings. Preferred heteroaryl groups include, but are not limited to, thienyl, furyl, indolyl, pyrrolyl, pyridinyl, pyrazinyl, oxazolyl, thiaxolyl, quinolinyl, pyrimidinyl, imidazolyl and tetrazolyl.
• Heterocycle or “heterocyclyl” means a 3- to 10-membered saturated or partially unsaturated non-aromatic monovalent cyclic radical having from one to 3 hetero atoms selected from nitrogen, oxygen or sulfur or a combination thereof.
• preferred heterocycles are piperidine, piperazine, pyrrolidine, and morpholine.
• IC 5 o refers to the concentration of a particular compound according to the invention required to inhibit 50% of a specific measured activity. IC50 can be measured, inter alia, as is described in Example 26, infra.
• “Pharmaceutically acceptable ester” refers to a conventionally esterified compound of formula I having a carboxyl group, which esters retain the biological effectiveness and properties of the compounds of formula I and are cleaved in vivo (in the organism) to the corresponding active carboxylic acid.
• lower alkyl esters are the methyl, ethyl, and n-propyl esters, and the like.
• lower alkyl esters substituted with NR 41 R 42 are the diethylaminoethyl, 2-(4-morpholinyl)ethyl, 2-(4-methylpiperazin-1 -yl)ethyl esters, and the like.
• acyloxyalkyl esters are the pivaloxymethyl, 1 - acetoxyethyl, and acetoxymethyl esters.
• carbonate esters are the 1 - (ethoxycarbonyloxy)ethyl and 1 -(cyclohexyloxycarbonyloxy)ethyl esters.
• aminocarbonylmethyl esters are the N,N-dimethylcarbamoylmethyl and carbamoylmethyl esters.
• “Pharmaceutically acceptable salt” refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula I and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases.
• Sample acid-addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like.
• Sample base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethylammonium hydroxide.
• the chemical modification of a pharmaceutical compound (i.e. drug) into a salt is a technique well known to pharmaceutical chemists to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. See, e.g., H. Ansel et. al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th Ed. 1995) at pp. 196 and 1456-1457.
• “Pharmaceutically acceptable,” such as pharmaceutically acceptable carrier, excipient, etc., means pharmacologically acceptable and substantially non-toxic to the subject to which the particular compound is administered.
• substituted as in substituted alkyl, means that the substitution can occur at one or more positions and, unless otherwise indicated, that the substituents at each substitution site are independently selected from the specified options.
• the invention relates to compounds of formula
• R 1 is selected from lower alkyl, and lower alkyl substituted with OR 3 , NR 3 R 4 , S(O) n R 3 , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl;
• R 2 is selected from H, lower alkyl, and lower alkyl substituted with OR 5 , OC(O)R 5 , NR 5 R 6 , S(O) n R 5 , aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl;
• R 3 and R 4 are independently selected from
• R 5 and R 6 are independently selected from
• R 9 is H or lower alkyl; and n is O, 1 or 2;
• the groups NR 3 R 4 , NR 5 R 6 and NR 7 R 8 as defined above may include one or more ring heteroatoms in addition to the above-mentioned N.
• the total number of additional ring heteroatoms, that is in addition to the above-mentioned N depends on the particular ring system involved. Preferably, there are no more than 1 or 2 additional ring heteroatoms.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with OR 3 .
• R 3 groups include aryl, aryl substituted with halogen, and aryl fused to a heterocycle.
• Preferred halogen groups include Br, Cl and F.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with OR 3 .
• Preferred R 3 groups include heteroaryl and heteroaryl substituted with OR 7 .
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with NR 3 R 4 .
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with S(O) n R 3 , wherein R 3 is lower alkyl.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with cycloalkyl.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with substituted cycloalkyl.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with heterocycle.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with substituted heterocycle.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with heteroaryl.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl substituted with substituted heteroaryl.
• the invention relates to a compound of formula I wherein R 1 is lower alkyl. In another embodiment, the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with OR 5 wherein R 5 is lower alkyl substituted with NR 7 R 8 .
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with NR 5 R 6 .
• the invention relates to a compound of formula I wherein R 2 is lower alkyl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with one or more OH groups or one NR 5 R 6 group.
• the invention relates to a compound of the formula I wherein R 2 is lower alkyl substituted by OR 5 .
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with OC(O)R 5 .
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with S(O) n R 5 wherein R 5 is lower alkyl and n is 1 or 2. In another embodiment, the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with aryl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with substituted aryl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with cycloalkyl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with substituted cycloalkyl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with heterocycle.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with substituted heterocycle.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with heteroaryl.
• the invention relates to a compound of formula I wherein R 2 is lower alkyl substituted with substituted heteroaryl.
• the invention relates to a compound of formula I wherein R 2 is H.
• the invention relates to a compound of formula I wherein R 3 is H. In another embodiment, the invention relates to a compound of formula I wherein R 3 is lower alkyl.
• the invention relates to a compound of formula I wherein R 3 is lower alkyl substituted with aryl, aryl fused to a heterocycle or a substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or substituted heterocycle.
• the invention relates to a compound of formula I wherein R 3 is aryl.
• the invention relates to a compound of formula I wherein R 3 is aryl fused to a heterocycle or a substituted heterocycle.
• the invention relates to a compound of formula I wherein R 3 is substituted aryl.
• the invention relates to a compound of formula I wherein R 3 is heteroaryl.
• the invention relates to a compound of formula I wherein R 3 is heteroaryl fused to a heterocycle or a substituted heterocycle.
• the invention relates to a compound of formula I wherein R 3 is substituted heteroaryl.
• the invention relates to a compound of formula I wherein R 3 is heterocycle.
• the invention relates to a compound of formula I wherein R 3 is heterocycle fused to an aryl. In another embodiment, the invention relates to a compound of formula I wherein R 3 is cycloalkyl.
• the invention relates to a compound of formula I wherein R 3 is substituted cycloalkyl.
• the invention relates to a compound of formula I wherein R 3 is lower alkyl, heterocycle fused to an aryl, aryl, substituted aryl, or aryl fused to a heterocycle.
• the invention relates to a compound of formula I wherein R 4 is H.
• the invention relates to a compound of formula I wherein R 4 is lower alkyl.
• the invention relates to a compound of formula I wherein R 4 is lower alkyl substituted with aryl, aryl fused to a heterocycle or a substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or substituted heterocycle.
• the invention relates to a compound of formula I wherein R 4 is aryl.
• the invention relates to a compound of formula I wherein R 4 is aryl fused to a heterocycle or a substituted heterocycle.
• the invention relates to a compound of formula I wherein R 4 is substituted aryl. In another embodiment, the invention relates to a compound of formula I wherein R 4 is heteroaryl.
• the invention relates to a compound of formula I wherein R 4 is heteroaryl fused to a heterocycle or a substituted heterocycle.
• the invention relates to a compound of formula I wherein R 4 is substituted heteroaryl.
• the invention relates to a compound of formula I wherein R 4 is heterocycle.
• the invention relates to a compound of formula I wherein R 4 is heterocycle fused to an aryl.
• the invention relates to a compound of formula I wherein R 4 is cycloalkyl.
• the invention relates to a compound of formula I wherein R 4 is substituted cycloalkyl.
• the invention relates to a compound of formula I wherein R 5 is H.
• the invention relates to a compound of formula I wherein R 5 is lower alkyl.
• the invention relates to a compound of formula I wherein R 5 is lower alkyl substituted with NR 7 R 8 , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl. Most preferably R 5 is lower alkyl substituted with NR 7 R 8 .
• the invention relates to a compound of formula I wherein R 5 is lower alkyl substituted by one or more OR 7 .
• the invention relates to a compound of formula I wherein R 6 is H.
• the invention relates to a compound of formula I wherein R 6 is lower alkyl.
• the invention relates to a compound of formula I wherein R 6 is lower alkyl substituted with NR 7 R 8 , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl. Most preferably R 6 is lower alkyl substituted with NR 7 R 8 .
• the invention relates to a compound of formula I wherein R 7 is H.
• the invention relates to a compound of formula I wherein R 7 is lower alkyl.
• the invention relates to a compound of formula I wherein R 8 is H.
• the invention relates to a compound of formula I wherein R 8 is lower alkyl.
• the invention relates to a compound of formula I wherein R 8 is H.
• the invention relates to a compound of formula I wherein R 8 is lower alkyl.
• the invention relates to a compound of formula I wherein R 9 is H. In another embodiment, the invention relates to a compound of formula I wherein R 9 is lower alkyl.
• the invention relates to a compound of formula I
• R 1 is lower alkyl substituted with OR 3 ;
• R 2 is H or lower alkyl substituted with one or more OR 5 groups or one NR 5 R 6 group;
• R 3 is aryl substituted with halogen or OR 7 , or is aryl fused to a heterocycle;
• R 5 and R 6 are independently H, lower alkyl or lower alkyl substituted by one or more OR 7 , or alternatively, the group NR 5 R 6 independently can form a ring having a total of from 3 to 6 atoms, said ring atoms comprising in addition to the nitrogen to which R 5 and R 6 are bonded, carbon ring atoms, said carbon ring atoms optionally being replaced by one additional heteroatoms selected from N or O, and said ring atoms optionally being substituted by OR 7 ; and
• R 7 is H or lower alkyl; or a pharmaceutically acceptable salt or ester thereof.
• the compounds of the invention are selective for FGFR and/or KDR kinases. These compounds are useful in the treatment or control of cancer, in particular the treatment or control of solid tumors, specifically breast, lung, colon and prostate tumors. These compounds are highly permeable to cell membranes and thus possess advantageous bioavailability profiles such as improved oral bioavailability.
• the compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds are provided in the examples. Generally, compounds of formula I can be prepared according to the below described synthetic routes.
• isomeric structures of formula I can be carried out according to known methods such as for example resolution or chiral high pressure liquid chromatography (also known as chiral HPLC). Resolution methods are well known, and are summarized in “Enantiomers, Racemates, and Resolutions” (Jacques, J. et al. John Wiley and Sons, NY, 1981 ). Methods for chiral HPLC are also well known, and are summarized in “Separation of Enantiomers by Liquid Chromatographic Methods” (Pirkle, W. H. and Finn, J. in “Asymmetric Synthesis", Vol. 1 , Morrison, J. D., Ed., Academic Press, Inc., NY 1983, pp. 87-124).
• the optional conversion of a compound of formula I that bears a basic nitrogen into a pharmaceutically acceptable acid addition salt can be effected by conventional means.
• the compound can be treated with an inorganic acid such as for example hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or with an appropriate organic acid such as acetic acid, citric acid, tartaric acid, methanesulfonic acid, p-toluene sulfonic acid, or the like.
• the optional conversion of a compound of formula I that bears a carboxylic acid group into a pharmaceutically acceptable alkali metal salt can be effected by conventional means.
• the compound can be treated with an inorganic base such as lithium hydroxide, sodium hydroxide, potassium hydroxide, or the like.
• the optional conversion of a compound of formula I that bears a carboxylic acid group or hydroxy group into a pharmaceutically acceptable ester can be effected by conventional means.
• the conditions for the formation of the ester will depend on the stability of the other functional groups in the molecule to the reaction conditions. If the other moieties in the molecule are stable to acidic conditions, the ester may be conveniently prepared by heating in a solution of a mineral acid (e.g., sulfuric acid) in an alcohol.
• Other methods of preparing the ester which may be convenient if the molecule is not stable to acidic conditions include treating the compound with an alcohol in the presence of a coupling agent and in the optional presence of additional agents that may accelerate the reaction. Many such coupling agents are known to one skilled in the art of organic chemistry.
• dicyclohexylcarbodiimide and triphenylphosphine /diethyl azodicarboxylate.
• the reaction is conveniently carried out by treating the acid with the alcohol, dicyclohexylcarbodiimide, and the optional presence of a catalytic amount (0-10 mole%) of N,N-dimethylaminopyhdine, in an inert solvent such as a halogenated hydrocarbon (e.g., dichloromethane) at a temperature between about 0 degrees and about room temperature, preferably at about room temperature.
• a halogenated hydrocarbon e.g., dichloromethane
• the reaction is conveniently carried out by treating the acid with the alcohol, triphenylphosphine and diethyl azodicarboxylate, in an inert solvent such as an ether (e.g., tetrahydrofuran) or an aromatic hydrocarbon (e.g., toluene) at a temperature between about 0 degrees and about room temperature, preferably at about 0 degrees.
• an inert solvent such as an ether (e.g., tetrahydrofuran) or an aromatic hydrocarbon (e.g., toluene)
• the present invention includes pharmaceutical compositions comprising at least one compound of formula I, or a pharmaceutically acceptable salt or ester thereof and a pharmaceutically acceptable excipient and/or carrier.
• compositions can be administered orally, for example in the form of tablets, coated tablets, dragees, hard or soft gelatin capsules, solutions, emulsions or suspensions. They can also be administered rectally, for example, in the form of suppositories, or parenterally, for example, in the form of injection solutions.
• compositions of the present invention comprising compounds of formula I, and/or the salts or esters thereof, may be manufactured in a manner that is known in the art, e.g. by means of conventional mixing, encapsulating, dissolving, granulating, emulsifying, entrapping, dragee-making, or lyophilizing processes.
• These pharmaceutical preparations can be formulated with therapeutically inert, inorganic or organic carriers. Lactose, corn starch or derivatives thereof, talc, stearic acid or its salts can be used as such carriers for tablets, coated tablets, dragees and hard gelatin capsules.
• Suitable carriers for soft gelatin capsules include vegetable oils, waxes and fats.
• Suitable carriers for the manufacture of solutions and syrups are water, polyols, saccharose, invert sugar and glucose.
• Suitable carriers for injection are water, alcohols, polyols, glycerine, vegetable oils, phospholipids and surfactants.
• Suitable carriers for suppositories are natural or hardened oils, waxes, fats and semi-liquid polyols.
• the pharmaceutical preparations can also contain preserving agents, solubilizing agents, stabilizing agents, wetting agents, emulsifying agents, sweetening agents, coloring agents, flavoring agents, salts for varying the osmotic pressure, buffers, coating agents or antioxidants. They can also contain other therapeutically valuable substances, including additional active ingredients other than those of formula I.
• the compounds of the present invention are useful in the treatment or control of cell proliferative disorders, including prevention of the formation of new blood vessels in solid tumors (anti-angiogenesis).
• anti-angiogenesis new blood vessels in solid tumors
• These compounds and formulations containing said compounds are particularly useful in the treatment or control of solid tumors, such as, for example, breast, colon, lung and prostate tumors.
• a therapeutically effective amount of a compound in accordance with this invention means an amount of compound that is effective to prevent, alleviate or ameliorate symptoms of disease or prolong the survival of the subject being treated. Determination of a therapeutically effective amount is within the skill in the art.
• the therapeutically effective amount or dosage of a compound according to this invention can vary within wide limits and may be determined in a manner known in the art. Such dosage will be adjusted to the individual requirements in each particular case including the specific compound(s) being administered, the route of administration, the condition being treated, as well as the patient being treated. In general, in the case of oral or parenteral administration to adult humans weighing approximately 70 Kg, a daily dosage of about 10 mg to about 10,000 mg, preferably from about 200 mg to about 1 ,000 mg, should be appropriate, although the upper limit may be exceeded when indicated. The daily dosage can be administered as a single dose or in divided doses, or for parenteral administration, it may be given as continuous infusion.
• the compounds of this invention may be used in combination (administered in combination or sequentially) with known anti-cancer treatments such as radiation therapy or with cytostatic or cytotoxic agents, such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase Il inhibitors such as etoposide: topoisomerase I inhibitors such as CPT- 1 1 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or epothilones; hormonal agents such as tamoxifen; thymidilate synthases inhibitors, such as 5- fluorouracil; and anti-metabolites such as methotrexate.
• cytostatic or cytotoxic agents such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase Il inhibitors such as etoposide: topoisomerase I inhibitors such as CPT- 1 1 or to
• the above-described combination products include the compounds of this invention within the dosage range described above and the other pharmaceutically active agent or treatment within its approved dose range.
• an early cdk1 inhibitor olomucine has been found to act synergistically with well known cytotoxic agents in inducing apoptosis. (J. CeII ScL, 1995, 108, 2897-2904).
• Compounds of formula I may also be administered sequentially with known anticancer or cytotoxic agents when concomitant administration or a combination is inappropriate. This invention is not limited in the sequence of administration: compounds of formula I may be administered either prior to or after administration of the known anticancer or cytotoxic agent.
• the cytotoxic activity of the cdk inhibitor flavopiridol is affected by the sequence of administration with anticancer agents. ⁇ Cancer Research, 1997, 57, 3375).
• the present invention is also directed to the following novel intermediates useful in the synthesis of compounds of formula I:
• Method A A mixture of 3-(4-methyl-thiophen-2-yl)-acryloyl azide (69.21 g, 0.358 mol) (from Intermediate 3 supra) and xylene (700 ml_) was stirred and heated at reflux for 0.5 hour. Iodine (0.45 g, 1.79 mmol) was added and mixture was heated at reflux over night. Reaction mixture was cooled and stirred for 5 minutes with aqueous sodium bisulfite solution. The suspension was filtered, washed with ether and sucked dry to give 3-methyl-5H-thieno[3,2-c]pyridin-4-one as a tan solid. (Yield 31.28 g, 52.8%).
• Method B 3-(4-Methyl-thiophen-2-yl)-acryloyl azide (1.54 g; 7.95 mmol) (from Intermediate 3 supra) was dissolved in meta-xylenes (16 ml_). The solution was heated in an oil bath at 105 - 115 0 C for 30 minutes until nitrogen evolution ceased. At this point a few crystals of iodine were added to the reaction and the oil bath temperature was increased to 145 - 150 0 C. The reaction was heated at reflux for 6 hours. Upon cooling, solid precipitated out of solution. Filtration and drying yielded 3-methyl-5H-thieno[3,2-c]pyhdine-4-one. (Yield: 1.05 g; 80.1 %).
• ⁇ /, ⁇ /-Dimethylformamide (1.0 ml_, 12.86 mmol) was then added and the mixture stirred with heating at 70 0 C for 30 minutes.
• a second portion of N,N- dimethylformamide (0.5 ml_, 6.43 mmol) was added and the mixture was heated at 70 0 C for another 30 minutes.
• ice was added to the solution and the mixture was extracted with ethyl acetate.
• the organic extract was washed with water, saturated aqueous sodium bicarbonate solution, water and brine.
• the aqueous phases were back washed with ethyl acetate.
• the ethyl acetate solutions were combined, dried (sodium sulfate), filtered, and concentrated under reduced pressure.
• Method B Ammonia gas was bubbled into a solution of 4-chloro-3- phenoxymethyl-thieno[3,2-c]pyridine-7-carboxylic acid ethyl ester (0.95 g; 2.05 mmol) (from Intermediate 12 supra) in dry dioxane (21 ml_) in a pressure bottle for 15 minutes. The bottle was then capped and the solution was heated at 120 - 125 0 C. The reaction was monitored by liquid chromatographic analysis and recharged with ammonia after 15 hours. The reaction was stopped after 40 hours. The reaction mixture was concentrated. The residue was partitioned between dichloromethane and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated.
• Ci7 H 16 N 2 O 3 S M.W. 328.39 Ammonia gas was bubbled into a solution of 4-chloro-3-phenoxymethyl- thieno[3,2-c]pyridine-7-carboxylic acid ethyl ester (38 mg; 0.11 mmol) (from Intermediate 14 supra) in dioxane (2.4 ml_) in a pressure bottle for 30 minutes. The bottle was capped and the clear, colorless solution was heated in an oil bath at 115 - 125 0 C overnight.
• Benzyl chloroformate (Aldrich) (7.5 mL of a 50% solution in toluene, 52.5 mmol) was added dropwise and the mixture was stirred at room temperature for 18 hours. Excess reagent was quenched with methanol. Solvent was removed under reduced pressure. The residue was diluted with water, and the resulting solution was acidified to pH 1 (with dilute hydrochloric acid). This aqueous solution was washed with dichloromethane, then treated with excess sodium carbonate (to pH 10), and extracted with ethyl acetate (3 X 100 mL). The ethyl acetate layers were combined, dried (MgSO 4 ), and filtered.
• Aldrich Benzyl chloroformate
• Benzyl chloroformate (Aldrich) (2.62 ml_ of a 50% solution in toluene, 18.39 mmol) was added dropwise. The mixture was stirred at room temperature for 18 hours. Excess reagent was quenched with methanol, and the solvent was removed under reduced pressure. The residue was diluted with water, and the resulting solution was acidified to pH 1 (dilute hydrochloric acid), washed with dichloromethane, then treated with excess sodium carbonate (to pH 10), and extracted with ethyl acetate (3 X 50 ml_). Ethyl acetate layers were combined, dried (MgSO 4 ), filtered, and concentrated under reduced pressure.
• ⁇ /-Formyl-4-hydroxypiperidine was synthesized from 4-hydroxypiperidine (Aldrich) according to the literature procedure of Baker, W. R. et al. J. Med. Chem., 1992, 35, 1722 - 1734.
• the product-containing fractions were combined and freeze-dried.
• the freeze-dried material (as trifluoroacetic acid salt) was combined with comparable material from another experiment and dissolved in ethyl acetate.
• the trifluoroacetic acid salt was neutralized by washing with 1 N sodium hydroxide and then washing to neutrality with water and brine.
• the organic phase was dried and concentrated.
• the residue was recrystallized from ethyl acetate - hexanes to give 4-amino-3-(4-bromo-phenoxymethyl)-thieno[3,2- c]pyridine-7-carboxylic acid (2-diethylamino-ethyl)-amide. (Yield: 32.7 mg, 43.0% combined yield for two experiments).
• the amorphous solid (trifluoroacetic acid salt) was dissolved in ethyl acetate and neutralized with a 1 N sodium hydroxide wash. The organic phase was washed to neutrality with water and brine, dried over sodium sulfate and concentrated. The material was then recrystallized from ethyl acetate - hexanes to give 4-amino-3-(4-bromo- phenoxymethyl)-thieno[3,2-c]pyridine-7-carboxylic acid (4-pyrrolidin-1 -yl-butyl)- amide. (Yield 11.9 mg).
• This compound may be prepared in a manner analogous to the compound of Example 12b using the corresponding amide and methanesulfonic acid.
• Example 17 The compounds of Examples 17 - 24 unless specifically exemplified can be prepared in a manner analogous to the compounds of Examples 12a and 12b using the corresponding amines to form the appropriate compounds.
• Example 17
• the antiproliferative activity of the compounds of the invention is demonstrated below in Examples 26 and 27. These activities indicate that the compounds of the present invention are useful in treating cancer, in particular solid tumors, more particularly cancerous solid tumors of the breast, lung, prostate and colon, most particularly cancerous solid tumors of the breast and colon.
• kinase assays were conducted using an HTRF (Homogeneous Time Resolved Fluorescence) assay. This assay is described in A. J. KoIb et. al., Drug Discovery Today, 1998, 3(7), p 333.
• recombinant EEE-tagged KDR was activated in the presence of activation buffer (50 mM HEPES, pH 7.4, 1 mM DTT, 10% glycerol, 150 mM NaCI, 0.1 mM EDTA, 26 mM MgCI 2 , and 4 mM ATP).
• activation buffer 50 mM HEPES, pH 7.4, 1 mM DTT, 10% glycerol, 150 mM NaCI, 0.1 mM EDTA, 26 mM MgCI 2 , and 4 mM ATP.
• Kinase activity assays were performed in 96-well polypropylene plates (Falcon) with a total volume of 90 ⁇ l_ in each well. Each well contained 1 ⁇ M KDR substrate (Biotin-EEEEYFELVAKKKK), 1 nM activated KDR, and a test compound with one of 8 assay concentrations ranging from 100 ⁇ M to 128 pM (1 :5 serial dilution).
• the kinase activity assay was done in the presence of 100 mM HEPES, pH 7.4, 1 mM DTT, 0.1 mM Na 2 VO 4 , 25 mM MgCI 2 , 50 mM NaCI (from KDR stock solution), 1 % DMSO (from compound), 0.3 mM ATP (at K m concentration) and 0.02% BSA.
• the reaction was incubated at 37 0 C for 30 minutes.
• 72 ⁇ l_ of reaction mixture was transferred into a STOP plate containing 18 ⁇ L of revelation buffer (20 mM EDTA, 50 mM HEPES, pH 7.4, 0.02% BSA, 10 nM Eu-labelled anti-pY antibody (final cone.
• FGFR activity assays were carried out as described above for the KDR activity assay with the following differences.
• GST-tagged FGFR enzyme was activated at room temperature for 1 hour in the following activation buffer: 100 mM HEPES, pH 7.4, 50 mM NaCI, 20 mM MgCI 2 , and 4 mM ATP.
• the IC 50 value is the concentration of test compound that reduces by 50% the enzyme activity under the test conditions described.
• the compounds of the present invention have KDR IC 50 values less than 5 ⁇ M, preferably less than 1.5 ⁇ M, or FGFR IC 50 values less than 5 ⁇ M, preferably less than 2.5 ⁇ M. Most preferably, the compounds of the invention have KDR IC 50 values less than 1.5 ⁇ M and FGFR IC 50 values less than 2.5 ⁇ M .
• test compounds of this invention in cell- based assays was evaluated by BrdLJ assay using the BrdLJ kit (Roche Biochemicals 1 -647-229).
• Human umbilical vein endothelial cells (HLJVEC, Clonetics CC-2519) were cultured in EGM-2 (Clonetics CC-3162) medium and seeded at 10000 cells per well in a volume of 200 ⁇ l_ of EGM-2 (Clonetics CC- 3162) media in a 96-well flat bottom plates (Costar 3595) overnight.
• test compound at 10X test concentration in serum starvation medium with 2.5% DMSO was added to the appropriate wells.
• the control wells contained 20 ⁇ l_ of serum starvation medium with 2.5% DMSO. Plates were returned to the incubator for 2 hours.
• 20 ⁇ l_ of growth factors at 10X assay concentration diluted in serum starvation media, FGF at 50 ng per ml_, or VEGF (R&D systems 293-VE) at 200 ng per ml_ were added.
• the final concentration of FGF in the assay was 5 ng per ml_ and the final concentration of VEGF in the assays was 20 ng per ml_.
• the growth factor free control wells had 20 ⁇ l_ per well of serum starvation media with the same amount of BSA as the wells with growth factors. The plates were returned to the incubator for an additional 22 hours.
• the cells were labeled with BrdU (Roche Biochemicals 1 -647-229), by adding 20 ⁇ l_ per well of BrdU labeling reagent that has been diluted (1 :100) in serum starvation medium.
• the plates werejhen returned to the incubator for 4 hours.
• the labeling medium was removed by draining the medium onto paper towels.
• the cells were fixed and DNA denatured by adding 200 ⁇ l_ of fixation / denaturation solution to each well and incubating at room temperature for 45 minutes.
• the fixation / denaturation solution was drained onto paper towels and to each well was added 100 ⁇ l_ of anti-BrdU-POD and the wells were incubated for 2 hours at room temperature.
• the antibody solution was removed and the wells were each washed 3 - 4 times with 300 ⁇ l_ PBS.
• 100 ⁇ l_ of the TMB substrate solution was added to each well and the wells were incubated at room temperature for 5 - 8 minutes.
• the reaction was then stopped by adding 100 ⁇ l_ per well of 1 M phosphoric acid.
• the plates were read at 450 nm with reference wavelength of 650 nm.
• the percent inhibition for each test compound was calculated by subtracting the absorbency of the blank (no cells) wells from all wells, then subtracting the division of the average absorbency of each test duplicate by the average of the controls from 1.
• the IC 50 value is the concentration of test compound that inhibits by 50% BrdU labeling, and is a measure of inhibition of cell proliferation.
• the IC 5 O is determined from the linear regression of a plot of the logarith
• the compounds of the present invention have VEGF - stimulated HUVEC assay IC 50 values less than 3 ⁇ M, preferably less than 1.5 ⁇ M, or FGF - stimulated HUVEC assay IC 5 O values less than 5 ⁇ M, preferably less than 3.0 ⁇ M, even more preferably less than 2 ⁇ M. Most preferably, the compounds of the invention have VEGF - stimulated HUVEC assay IC 5 O values less than 1.5 ⁇ M and FGF - stimulated HUVEC assay IC 5 O values less than 2 ⁇ M.
• Compound A represents a compound of the invention.
• Compound A represents a compound of the invention.

Landscapes

• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Health & Medical Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Chemical & Material Sciences (AREA)
• Medicinal Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Applications Claiming Priority (1)

Publications (1)

Family

ID=35159670

Family Applications (1)

Country Status (9)

Families Citing this family (7)

Citations (1)

Family Cites Families (4)

• 2005
  • 2005-08-16 MX MX2008002060A patent/MX2008002060A/es not_active Application Discontinuation
  • 2005-08-16 WO PCT/EP2005/054022 patent/WO2007019884A1/en active Application Filing
  • 2005-08-16 CA CA002619459A patent/CA2619459A1/en not_active Abandoned
  • 2005-08-16 JP JP2008526383A patent/JP2009504692A/ja not_active Withdrawn
  • 2005-08-16 AU AU2005335661A patent/AU2005335661A1/en not_active Abandoned
  • 2005-08-16 KR KR1020087003603A patent/KR100995790B1/ko not_active IP Right Cessation
  • 2005-08-16 BR BRPI0520482-8A patent/BRPI0520482A2/pt not_active IP Right Cessation
  • 2005-08-16 EP EP05792209A patent/EP1917015A1/de not_active Withdrawn
• 2008
  • 2008-02-03 IL IL189181A patent/IL189181A0/en unknown

Patent Citations (1)

Also Published As

Similar Documents

Legal Events