JPH05507072A - Heterocyclic ethenediyl compounds that inhibit EGF receptor tyrosine kinase - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Invention of Heterocyclic Ethendiyl Compounds that Inhibit EGF Receptor Tyrosine Kinase field FIELD OF THE INVENTION This invention relates to the inhibition of cell proliferation. More specifically, the present invention In the inhibition of cell proliferation, including useful inhibitors of antityrosine kinases (PTKs) The present invention relates to the use of heterocyclic ethenediyl compounds.

Normal cell regeneration occurs when the cell matrix is injected with one or more growth factors, such as insulin, insulin, etc. Exposure to epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) It is believed that this is initiated by Such growth factors are typically Corresponding growth factor receptors embedded in or penetrating cell membranes This is unique to Pta. The initiation of cell regeneration is triggered by a corresponding reaction on the external surface of the cell membrane. It is thought to occur when a growth factor binds to a receptor. This growth factor Child-receptor binding changes the chemical characteristics of the relevant portion of the receptor and moiety is present within the cell and the receptor itself or called autophosphorylation. functions as an enzyme that catalyzes the phosphorylation of intracellular substrates. Such phospho Examples of lylating enzymes catalyze the phosphorylation of tyrosine amino acid residues in substrate proteins. This includes tyrosine kinases.

The pathophysiology of polymorphism is characterized by uncontrolled cell regeneration. It is found in many types of cells and is associated with leukemia, cancer, psoriasis, atherosclerosis and reincarnation. This includes stenotic injuries, etc. Inhibition of tyrosine kinases leads to uninhibited cell regeneration , that is, it is thought to be useful in suppressing cell proliferative diseases.

Self-phosphorylation, i.e. phosphorylation of the growth factor receptor itself and intracellular groups Initiation of phosphorylation in the host host is involved in mitogenesis and cell proliferation. It is a biochemical event. Insulin receptor autophosphorylation and other receptor Phosphorylation of substrates by septa is the earliest identifiable biochemical hormonal response. It is.

Insulin receptor and epidermal growth factor (EGF) receptor proteins Cells that respond to insulin and IGF production with synkinase (PTK) activity Loss of genetic material by site-directed mutagenesis results in complete loss of biological activity of the receptor. Resulting in total loss. This is completely undesirable. That is, in Surin is required by the body to perform other biological functions unrelated to cell growth. This is because it is something that will be done. Therefore, the surface area of the insulin receptor is inhibited. a compound that inhibits the claw portion of the EGF receptor at a concentration less than the concentration required to may provide valuable agents for the selective treatment of cell proliferative diseases.

reported facts Furanal malononitrile and chelate as tumor growth inhibitors The use of nylidene malononitrile compounds is malononitriles such as Gal (Cal). Studies on the biological effects of cal Action of Malononitrile) transplanted into 17 mice. The effect of substituted malononitriles on tumor growth The most potent inhibitor of fEGF receptor is EGF- inhibits induced proliferation, but inhibits such cells when induced by other growth factors. It has been reported to have little or no effect on cell proliferation. Also, Rubustatin (erbstatin) inhibits the EGF receptor within the membrane of A431 cells. inhibits self-phosphorylation of T. Low concentrations of erbstatin activate the EGF receptor. required for inhibition of self-phosphorylation, but higher concentrations of erbstatin Adenosine 3°, 5°-monophosphate (cAMP)-dependent protinoxin required to inhibit enzymes.

Summary of the invention According to the present invention, there is provided a method for inhibiting cell proliferation in patients suffering from the above-mentioned diseases. A method is provided, the method comprising: lylethendiyl or heteroarylethenediyl aryl compound or administering to said patient a pharmaceutically acceptable salt thereof; The aryl group may be a monocyclic or bicyclic heteroaryl group; monocyclic or bicyclic heteroaryl or bicyclic or tricyclic carbocyclyl The compound may optionally be mono- or polysubstituted. However, in the heteroaryl group, the ethenediyl group is a geminal cyano substituent. provided that it is not a furyl or chenyl group.

According to another aspect of the invention, a pharmaceutical composition is provided, the composition having a pharmaceutically acceptable a pharmaceutically effective amount of a compound of the above type in admixture with a carrier; It is characterized by

According to yet another aspect of the invention, novel compounds of the above type are provided.

Compounds of formula I below are a group of compounds of the above type used in the practice of this invention. Heteroarylethendiyl or heteroarylethenediyl aryl compounds Configure.

R3-C(W)=CR,R,1 where W is an about 5 to about 7 membered monocyclic ring containing 1 or 2 N, 0 or S atoms. ring or about 8 to about 12 members containing 1 to about 4 N, O or S atoms Represents a heteroaryl system having two cyclic rings, where the ring system optionally has 1 to Optionally substituted with about 3 groups R1, R1 is alkyl, -CN, -CON RRl-CSNRR or -CDOR, R is hydrogen, alkyl or aral R1 is a tricyclic arylcarboxyl ring system of -W, -CN, and the ring system may be optionally substituted with 1 to about 3 groups R4, R3 is hydrogen, alkyl, -CN　, -CH,CN, -CON肱-CSNRRl-CD OR or -CH=C(CN)CONH, and each R4 is independently an alkyl group. acetylamino , alkylthio, -CN, -CF3, nitro, -CDOR, -CONRR, -CSNRR, -C)10, -CH=C1(C00H, -NHCO(C)It) represents tcOOH or mopoly, R, is amino, -coNI (t, or represents hydrogen, alkyl, hydroxy, alkoxy or and R7 is independently alkyl, hydroxy, alkoxy, nitro, carbon. ruboxy or carbalkoxy, n is 0 to about 6, and m is 1 to about 7. Yes, but if both R1 and cFL are one CN, W is frill or Provided that it is not Chenil.

Compounds within the scope of the invention are substrates of the tyrosine kinase domain of the EGF receptor. It also shows specific affinity for the PDGF receptor kinase. inhibits the EGF receptor kinase and also inhibits the EGF receptor kinase. - Effectively inhibits dependent autophosphorylation.

Detailed description of the invention As used above and throughout this disclosure, the following terms are not otherwise stated) It should be understood that it has the following meaning.

"Alkyl" means a saturated aliphatic hydrocarbon having about 1 to about 6 carbons. It can be a straight chain or branched group containing atoms.

"Lower alkyl" means a group as defined above having from 1 to about 4 carbon atoms, directly Can be chained or branched, e.g. methyl, ethyl, propyl, isopropyl, butyl It can be tyl, isobutyl, see-butyl or tert-butyl.

"Alkoxy" means an alkyl-oxy group, which has the same meaning as "alkyl". It's like that. A lower alkoxy group is preferred. Examples are methoxy, ethoxy , n-propoxy, i-propoxy and n-butoxy.

"Aryl" means an unsaturated or partially unsaturated ring system, with preferred aryl Ryl groups are pyridyl and indolyl.

“Acyl” is an organic acid derived from an organic acid or carboxylic acid by removing its acidic hydroxyl group. A preferred acyl group is a lower alkyl carboxylic acid group, e.g. Examples are acetyl and propionyl groups. Also preferred is a benzoyl group.

"Halo" means halogen. Preferred halogens are chlorine, bromine and fluorine It is.

Preferred aralkyl groups are benzyl and phenethyl groups.

Therapeutically effective receptor tyrosine kinase (EGFRK) ) but not with adenosine triphosphate (ATP). It is being

PTK-inhibitors quercetin and genistein (gani) that compete with ATP stain) inhibits other protein kinases, resulting in cytotoxicity. It has a high value. As a test of selectivity, insulin receptor kinase (I ■) and/or EGFR better than inhibiting PDGF receptor kinase. Compounds that inhibit K are of great value.

The solubility of the compounds of the invention in both water and mildly hydrophobic solvents is It is theorized that this would increase the probability of permeation through the cell membrane. However, various defects Soluble compounds showed significant EGFRK inhibitory activity in in vitro tests.

A preferred group of compounds useful in the practice of this invention are compounds represented by Formula 1 above. includes. Here, W is a pentatom containing 1 or 2 N, 0 or S atoms. is a 6-membered monocyclic aryl ring, or contains 1 to 4 N, O or S atoms represents a 9- to 10-membered bicyclic aryl ring, which ring optionally has 1 to about 3 may be substituted with R4 groups, R, is -CN, -CONRRl-C3N represents RR or -COOR group, R represents hydrogen, alkyl or aralkyl, R2 is -W, -CN, -CONHR,, -CONRRl-COOR', -C 3NRR or , R, is a hydrogen atom, and each R4 is independently alkyl, hydride. roxy, alkoxy or halo, R3 represents amino, -CONHz, Each R6 is independently hydrogen or an alkyl group, and each R2 is independently an alkyl, hydro xy, alkoxy, halo, nitro, carboxy or carbalkoxy; n is O~4, m is 2~6, provided that both R1 and Pan2 are -CN. , provided that W is not a frill or a chenille.

A more preferable compound of the present invention is a compound of the above formula 1 in which W is furyl, pyrrolyl, or Chester. Nil, thiazolyl, pyridyl, imidazolyl, isoimidazolyl, pyridazinyl , pyrimidinyl, benzofuranyl, indolyl, indolinyl, indolinonyl , benzothenyl, benzothiazolyl, quinolinyl, isoquinolinyl, chromeni 1,3-benzodioxolyl or 2,3-dihydro-1,4-benzodiolyl Includes compounds that are xynyl.

The most preferred compound is the above general formula I, where W is pyridyl, indolyl, imidolyl, Dazolyl, benzothiazolyl, 1,3-benzodioxolyl or 2,3-dihydryl is dolo-1゜4-benzodioxinyl, R1 is -CN, -CONRRl-C 3 substituents or -COOR group, R represents hydrogen, alkyl or aralkyl, R2 is -W, -CN, -coN'HRs, -CONRRl-COOR, -C SNRR or , R1 is a hydrogen atom, and each R4 is independently lower alkyl, hydroxy, lower alkoxy or halo, and R8 is represents, and each R7 independently represents lower alkyl, hydroxy, alkoxy, halo, nitro , carboxy or carbalkoxy, n is O~4, and m is 2~ 6 is included.

Compounds of the invention are useful as free bases, salts and hydrates. all of these Any shape is within the scope of the invention. Acid addition salts can be formed, but are simply the most convenient It is only an advantageous form of use, and in reality, the use of the salt form is ultimately inferior to the use of the base form. Equals use. Acids that can be used to prepare acid addition salts preferably include the free base and Pharmaceutically acceptable salts when combined, i.e. the anion is the pharmaceutical dosage of the acid salt. is harmless to animal organs and as a result the inherent advantageous properties of the free base are Includes those that form salts that are not impaired by side effects attributed to anions. do. Although pharmaceutically acceptable salts of the basic compound are preferred, certain salts themselves may Even if the acid salt is desired only as a product, for example for purification and identification purposes. or if the acid salt is formed only by ion exchange techniques, it is pharmaceutically acceptable. as a source of the free base form when used as an intermediate in the preparation of salts. Any acid addition salt is useful.

Pharmaceutically acceptable salts within the scope of this invention include the following acids, such as hydrochloric acid, sulfuric acid, phosphoric acid. and inorganic acids such as sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malon acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluene Derived from organic acids such as sulfonic acid, cyclohexylsulfamic acid, and quinic acid Includes salt.

Corresponding acid addition salts are hydrochloride, sulfate, phosphate, and sulfamate, respectively. , acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate phosphate, benzenesulfonate, l)-toluenesulfonate, cyclohexyl Contains sulfamate and quinate.

Acid addition salts of the compounds of this invention can be prepared by converting the free base into an aqueous or aqueous solution containing a suitable acid. alcohol solution or other suitable solvent and then evaporate the solvent. or by reacting the free base with an acid in an organic solvent. It can be prepared by In the latter case, the acid salt can be separated directly or The acid salt can be obtained by concentrating the solution.

Compounds useful in the practice of this invention can be prepared using known methods such as U.S. Pat. It can be prepared by Nebenagel condensation reaction as described in No. 9゜148. .

The compound of the present invention can be prepared according to the following reaction formula: R$-C(W)O+ Rt- CHt-Rt -> R3-C(W) = CR, Rz In polar medium, ammonia or or in the presence of an amine, such as piperidine, at elevated temperature, of the formula λrcHt-R2 Kunevener of the heterocyclic aldehyde of formula W with an active methylene compound of Gel condensation gives the products of the invention. If substitution of the group R4 is desired, the corresponding A ketone starting material is used. Reaction temperature and reaction within the range of 25°C to reflux temperature The time will vary depending on the materials used in the condensation.

The compounds of the present invention are commercially available, known in the literature, or known in the literature. It can be manufactured by the following method.

Various substituents R, L, R1, Rs, R,, Rs, on the heteroring or mirror Rs and T can be present in the starting compound and after formation of the condensation product, one It can be added by a known method for substituting or converting a group into another group. Appropriate location If the substituent itself is reactive, the substituent itself may be modified according to techniques known in the art. Can be protected.

Green's ``Protective Groups in Organic Synthesis'' Groups in Organic Synthesis), 1981, Jo Can be found at Nwilly & Sons. For example, to nitrate a nitro group to an aromatic ring and convert the nitro group to another group, e.g. by reduction, into a halo group by diazotization of the amino group and substitution of the diazo group. can. An acyl group can be substituted onto an aryl group by Friedel-Crafu acylation. Wear. The acyl group is then subjected to Borchukishner reduction and Clemensen reduction. can be converted to the corresponding alkyl group by various methods including. Alkyl amino group can be converted to form mono- and di-alkylamino groups, mercapto and hydride. Roxyl groups can be alkylated to form the corresponding ethers. primary alcohol It can be oxidized with oxidizing agents known in the art to form carboxylic acids or aldehydes.

Also, secondary alcohols can be oxidized to form ketones. Thus, replacement or Use exchange reactions to add substituents to entire molecules of starting materials, intermediates, or final products. can give.

Compounds within the scope of the invention have significant activity as protein tyrosine kinase inhibitors. and for the treatment of several conditions including, for example, psoriasis and restenotic lesions. It has therapeutic value as a cell growth agent. The present invention is particularly applicable to atherosclerosis It is expected that it will be applicable to the treatment of Some conditions, e.g. atheromatous arteries In connection with the treatment of sclerosis, some people may, for example, consider genetic, environmental or medical history factors. It may be considered that there is a high risk for this reason. Compounds within the scope of this invention are to prevent or delay the occurrence or recurrence of a condition, or to treat the condition. It can be used to

The compounds of the invention may be administered in species that are compatible with the chosen route of administration, e.g. oral or parenteral administration. It can be administered to a mammalian host in various forms. Parenteral administration here refers to intravenous, intramuscular administration. Transepithelial administration including intravenous, subcutaneous, intraocular, intrasynovial, transdermal, intraocular, sublingual and buccal etc., as well as topical routes of administration by inhalation and aerosol; bulbar, rectal and nasal inhalation and rectal systemic route )including.

The active compound may be present, for example, with an inert diluent or with an assimilable edible carrier. Both can be administered orally or can be enclosed in hard or soft shell Seratin capsules. Alternatively, it can be compressed into tablets, or even added directly to therapeutic food. Oral therapeutic dosage For administration, the active compound may be formulated with excipients to form orally ingestible tablets, buccal tablets, etc. As tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. Can be used. Such compositions or formulations contain at least 0.1x the active ingredient Should. The proportions of active ingredients in the compositions and formulations can be varied and advantageously may range from about 2% to about 6% of the weight of the unit. Such therapeutically useful The amount of active ingredient in the composition is such that a suitable dosage will be obtained. Advantages of the present invention Preferred compositions or formulations include oral dosage units containing about 1 to 1000 mg of active ingredient. It is prepared to contain.

The tablets, troches, pills, capsules, etc. may also contain the following ingredients: . That is, binders such as gum tragacanth, gum acacia, corn starch or or gelatin; excipients such as dicalcium phosphate 2 disintegrants, such as corn star lubricants, such as magnesium stearate, potato starch, alginic acid, etc. ; and sweeteners such as sucrose, lactose or saccharin may be added. You can also add aromatics such as peppermint, oil of wintergreen or cherry flavor. good. If in dosage unit form or capsule, it may contain, in addition to the above type of ingredients, a liquid carrier. can be included. Various other materials can act as coatings or modify the physical structure of the dosage unit. Can exist to improve shape. For example, tablets, pills or capsules are It can be coated with water, sugar, or both. syrup or elixir The active compound, sucrose as a sweetener, methyl as a preservative and Contains propylparaben, dyes and fragrances such as cherry or orange flavor. You can Any material used in the preparation of any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts used . Additionally, the active compounds can be formulated into sustained release preparations and formulations.

The active compounds can also be administered parenterally or intraperitoneally. Free base also prepares a solution of the active compound as a pharmaceutically acceptable salt in hydroxypropylcell. It can be prepared as an aqueous solution mixed appropriately with a surfactant such as loin. glycerol , liquid polyethylene glycol, mixtures thereof, and dispersants dispersed in oil are also prepared. can. Under normal conditions of storage and use, these preparations inhibit the growth of microorganisms. Contains preservatives to prevent

Dosage forms suitable for injectable use are sterile aqueous solutions or dispersions, or sterile injectable solutions or Includes sterile powders for extemporaneous preparation of dispersions. In all these cases The dosage form should be sterile and fluid to the extent that easy syringability exists. Should. It must be stable under the manufacturing and storage conditions and must be stable under the conditions of manufacture and storage. must be protected against the contaminating action of microorganisms such as bacteria and fungi. carrier For example, water, ethanol, polyols (e.g. glycerol, propylene) liquid polyethylene glycol, etc.), suitable mixtures thereof, and It may be a solvent or dispersion medium containing oil or the like. Appropriate fluidity is determined by e.g. In the case of dispersants, maintaining a specified particle size, and using surfactants, etc. It can be maintained depending on the purpose. Suppression of microbial action is achieved using various antibacterial agents and antifungal agents. Fungicides, such as parabens, chlorobutanol, phenol, sorbic acid, thimero This can be achieved by Searle et al. In many cases, isotonic agents such as sugars or chlorides It is desirable to include sodium. Prolonged absorption of injectable compositions may result in delayed absorption. obtained by the use of spreading agents such as aluminum monostearate and gelatin. I can do it.

Sterile injectable solutions contain a predetermined amount of the active compounds along with various other ingredients enumerated above. Both are prepared by blending in a suitable solvent and then sterilizing by filtration. . Generally, dispersants involve incorporating various sterile active ingredients into a sterile vehicle. The vehicle is prepared by a basic dispersion medium and an appropriate selection from those listed above. Contains other necessary ingredients. For sterile powders for the preparation of sterile injectable solutions, preferred The preparation method is vacuum drying and freeze-drying, and the method uses pre-sterilized and filtered solutions. provides a powder of the active ingredient and any additional prescribed ingredients.

The therapeutic compounds of this invention, alone or in combination with pharmaceutically acceptable carriers such as those described above, administered to mammals in parallel. The compounding ratio of the compound depends on its solubility and chemical properties. determined by the quality, route of administration chosen and standard pharmaceutical practice.

The dose of the therapeutic agent of the present invention that is most suitable for prevention or treatment depends on the mode of administration, the selected will vary depending on the particular compound and the physiological characteristics of the particular patient being treated. . In general, use lower doses initially and, if necessary, to optimize efficacy under the circumstances. Gradually increase in small amounts until . According to physiological studies using rats, humans The therapeutic dose of phthalate is generally about 0.01 to about 100 μg/kg per day (body weight). weight) or about 0.4g to about 10g or/and more, but per day It can be administered as a dosage unit in one to several different doses. Oral administration is higher Requires large doses.

Example Embodiments of the invention are described in the following non-limiting examples. This example applies to humans and other animals. Also includes a description of pharmacological testing methods that are thought to correlate with therapeutic activity between . Examples 1-11 below are illustrative of compounds within the scope of this invention. Examples 1-5 , various unsubstituted heteroarylmalononitriles, i.e., R+ and corners are respectively -CN and all other substituents R are hydrogen atoms. Example 6, R1 is -CN, R7 is -C(Nl2)=C(CN)2, A compound in which R is hydrogen and has no R4 substituent is exemplified. In Example 7, , nitro-substituted heteroarylmalononitriles, i.e. R, and section 2 are each - A compound in which R1 is CN, R1 is hydrogen, and R4 is a nitro group is exemplified. implementation In Example 8, the diheteroarylethendiyl compound, i.e., R1 is -CN and R Examples of compounds in which 2 is a pyridyl group and all other substituents R are hydrogen atoms Ru. In Example 9, the heteroarylethendiyl compound, that is, R9 is a hydrogen atom, , R1 is a phenanthryl group, R8 is -CN, and the R4 substituent is The following is an example of a compound that has no properties at all. In Examples 1O and 11, the heteroaryl group a tendiylaryl compound, that is, R1 is -CN and R2 is 3,4-silonito; A compound in which R1 is -H and is a roxybenzoyl group is exemplified.

Example 1: 0.4g (4mM) of 2-imidazolidenemalononitrile limidazole (Aldrich) and 0°5 g (4,5 rfM) of malononitrile in 15 ml of ethanol, 30 mg of β-alani was added. The reaction system was refluxed for 1 hour, evaporated to dryness, and the resulting dark solid was Flash chromatography on silica gel (eluent: CHtC1t/3 % ethanol) to give a yellow color with a melting point (ILp, ) of 180°C (decomposition). 0.2 g of solid (yield 3x) was obtained.

Example 2.5-Vyrazolidenmalononitrile (a) CLClt (10g; die 2.5 g (IS) in 50 ml ether solution of , 5-) of propylene acecool was added. After freezing for 3 days, 50ml of eta Nord and 10 drops of concentrated MCI were added and the reaction mixture was stirred at room temperature for 15 minutes. evaporated to dryness and crushed using CC1 to give 90 [formylpyrazole]. Obtained as a pale yellow solid.

(b) 80 μ of the pyrazole from step (a) above and 70 μ of malonitrile are combined into 2 1) Add 2-β-alanine to a solution dissolved in ml of ethanol, and add it to the reaction. The reaction mixture was refluxed for 20 hours. Water was added and the reaction mixture was extracted with F,tAc. , evaporated, flash chromatographed, ILI), 1.54°C The product was produced as a yellow solid (yield 254-hydroxyimidazole (aldo (Aldrich)) with NOx and the reaction mixture was evaporated to dryness. 0.5 g of a light brown solid was obtained. Add this to 10ml of ethanol and 2ml 0.45 g (6.8 mM) of malononitrile and and 50 μl of β-alanine were added. The reaction mixture was refluxed for 1 hour and evaporated to dryness. and flash chromatography. l1l) from the second fraction, 0.8g (6.7m) of yellow solid indoline and ethoxymethylene malononi at 176℃ A solution of 0.8 g (6.7 m) of tolyl dissolved in 30 ml of CH, CN was brought to room temperature. Stir for 30 minutes, filter the formed precipitate, m, p, pale yellow solid at 228 ° C. Product o, 7 gg (yield SI%) was obtained.

Example 5: 150 mg (lrrM) of 5-indribunemalononitrile Milindole and 80■(1,2-) malononitrile were added to 10ml of ethanol. One drop of piperidine was added to the solution dissolved in the solution. This reaction solution was heated for 2.5 hours. Reflux, add H2O, cool the resulting mixture and filter to m, p, 225 140 ml of product was obtained as a yellow solid at 70% yield.

Example 6: 1.1.3-tricyano-2-amino-4-(5-indole)buta Gen0, 29 g (2 dl) of 5-formylindole and 0.28 g (2,1 m M) malonitrile dimer dissolved in 10 ml of ethanol, 3 0 μ of β-alanine was added. The reaction mixture was refluxed for 4 hours, cooled and filtered. washed with water and ethanol and dried (ffLI), yellow-orange at 242°C. 0.16 g (31% yield) of product was obtained as a solid.

Example 7: 3.4-methylenedioxy-6-nidrobenzylidenemalononitrile 3.4-methylenedioxy-6-nitrobenzaldehyde of Ig (5,1 bacteria), 0.4g (>6mM) malononitrile and 0.2g β-alanine in 30m A solution of 1 in ethanol was stirred at room temperature for 16 hours. Add 50ml of water added. Filtered to give 1 g of product as a bright yellow solid, ip, 104°C ( A yield of 80% was obtained.

Example 8 2-(2-pyridyl)-3-[3,4-(methylene dioxy)phenyl]-2-propenenitrile was prepared. Melting point is 153-15 The temperature was 5°C.

Example 9: 2-(4-pyridyl)-3-(9-phenanthryl)-2-propene Phenanthrene-9-carboxalde of nitrile 0,5g (2,43dd) Add 0.375 g (2.43 m ) of 4-pyridylacetonitrile HCI and 0.5 ml of piperidine were added. This reaction mixture was stirred and heated under reflux for 2 hours, at which point a yellow precipitate appeared. Formed. The reaction mixture was heated to reflux for an additional 2 hours, cooled to room temperature, and the solvent was partially removed. Evaporated in portions. This mixture was partitioned between CI (,C1,,!:H20) , washed with 5% NaHCO, solution, dried over anhydrous sodium sulfate, filtered, Concentration gave a yellow solid, which was then flash chromatographed on silica gel. I applied it to the graph. This material was placed on a column containing CH, CI, and EtO Elute with Ac:hexane (1:l), m, p, amorphous at 206-208℃ 0.328 g (44x yield) of product was obtained as a yellow solid.

5-formylindole (145■; 1mM), 3.4-dihydroxybeta lnzoylacetonitrile (177■; In!il) and β-alanine (30■ ) in 30 ml of ethanol was refluxed for 5 hours. Add water and get The reaction mixture was extracted with EtOAc.

The organic phase is concentrated and the resulting oily residue is chromatographed on silica gel. The title compound was purified as an orange solid at m-p, 185°C (yield: 31 x) was obtained.

3-formylindole (105■, 0.7m), 3,4-dihydroxyhe cylacetonitrile (130 dJ, 0.7 dJ) and β-alanine (20 dJ) ) in 30 ml of ethanol was refluxed for 5 hours. Add water and get The reaction mixture was extracted with EtOAc. Concentrate the organic phase and the resulting oily solid C)1. Crushed with CI2 and marked as an orange solid at m, p, 228°C. 160 mg (yield 72%) of the compound was obtained.

Compounds of the present invention are subjected to various biological tests, and the results are not useful. Correlates with reproductive activity. These tests test the EGF receptor of the compounds described herein. Ptakinase, PDGF receptor kinase and insulin receptor kinase Useful for determining inhibitory activity.

Purification of EGF-receptor A method for purifying EGF-receptors was described by Yarden and Schlessinger. A, nd Schlessinger) method. A431 cells are grown in 80 an' confluent culture bottles (2XIO' cells/bottle). this Wash the cells twice with PBS, 1. Harvest in PBS containing OmM EDTA (3 1 hour at 7°C) and centrifuged at 600g for 10 minutes. Cut this cell twice07 1. per cell. ml cold solubilization buffer (50111'11 Hepa s) Buffer, p) I 7.6; 1°C % Triton -100; 150n1111NaCI; 5m EGTA + 1mM PlS F;50 gg/ml aprotinin, 25-benzamidine, 5 μg/ml I leupepti Solubilize for 20 minutes at 4°C in do. After centrifuging at 100,000 g for 30 minutes, the supernatant was transferred to EA-agarose. onto a column (packed with 100 μl of resin per 2×10′ cells) at 4°C. Shake for 2 hours. Remove unabsorbed substances and cleanse the resin) buff y (50-Hepes , pH 7,6; 0.1% dryide:/X-1oo: 150mM NaCI ), twice with HTN buffer containing 1M NaCl, and mG buffer. (50mM Hepes, pH 7,6; 0.L% Triton X-100; Wash twice with 150 mM NaCl and 10% glycerol). This E GF-receptor was treated with 0.5!11 N-acetyl-D-glucose in a batchwise manner. Elute with HTNG buffer containing min (200 μl per 2X10' cells) do.

The eluted material was stored in aliquots at -70°C and treated with TMING prior to use. Buffer (50rrM Trls-Mes buffer y-, pH 7.6; 0. 1% Triton X-100; 150 Inlll NaCl and 10% glycero dilute with water

EGFR kinase-catalyzed phosphorylation of poly(GAT) and its inhibition W GA-purified EGFR (0.25 μg/assay) was crystallized for 20 min at 4°C. Preactivation with EGF (0,85 μM) in ris-Mes buffer, pH 7,6 become The assay consists of Mg(Ac)2 (60 reagents), [γ-”PIATP (125 gM, 2-5 μCi/assay), Boba GATX 0.0625 mg/ml. 0.125 mg/m! , 0.25111 g/ml) and 2 of each of 6 concentrations. Begin by adding a mixture containing several inhibitors. The temperature for this assay was 22°C. It can be seen that the formation of phosphorylated polymer is linear up to 20 minutes.

The PTK inhibitors tested were tested so that the final concentration of ethanol did not exceed 4x during the assay. Soluble in water or a mixture of ethanol and water. Ethanol up to 4% Its presence in the assay has no effect on EGFR kinase activity. Applicable The concentration of EGF in the assay is 300 nM in a final volume of 40 μl. 5.10 or after 20 minutes, aliquots of 25 μl were cut into 3-an pieces of Fattman filter paper. and then in cold 10 STCA containing 0.01 M sodium pyrophosphate. Soak. - After washing at 4°C overnight, dry the cut filter paper, and! 2P Cherenkov (Cerenkov) Count by measuring radiation. Concentration poly(GAT ) dependence is Michaelian, GlusAl, a, Calculated per 5Tyr (GAT) unit = 0.076±0, 007■/m l or 0.069±0.007-. For poly(GAT) phosphorylation This EGF response is shown graphically. - for ATP in this assay is 2 ．． It was found to be 9 μM. The compounds of the above examples were added to this EGF-receptor. It was assayed for its ability to inhibit tyrosine kinase activity. According to Table 1 below. The results are summarized. rics. ” is a poly(GAT) phosphor under standard assay conditions. It represents the concentration (μM amount) of the inhibitor required for 50% inhibition of sphorylation. PolyGAT The concentration was equivalent to a 2.3-value.

The above results of this assay demonstrate that the compounds of the invention are EGFRK-catalyzed substrates. It has been shown to be effective in inhibiting phosphorylation.

Time dependence of autophosphorylation of EGF-receptor WGA from A431 cells Purified EGF-receptor (0.5 μg/assay) was incubated at 4° C. for 20 min [! GF (800 nM). This reaction consists of Mg(Ac)t (60ml), T ris-Mes buffer, p) 17, 6 (50 rewards) and [”PIATP (20 Begin by adding MM: 5 μCi/assay).

The reaction was carried out at 4°C or 15°C, and sodium dodenyl sulfate (SDS) assay was performed. Food buffer y (10% glycerol, 50dσris, pH 6, 8, 5% β- Stop by adding mercaptoethanol and 3% SO3”). 8%-3DS polyacrylamide gel (SDS-PAGEX 30% acrylic prepared from amide and 0.8 bis(acrylamide) with 0.375M T ris; pH 8, 8, 0, 1% SOS, 0.05% TEMED and 0.4 6% ammonium persulfate). Dry this gel and automate Radiography with Agfa Curix RP2 - Performed using line film.

The relevant radioactive band portions are cut out and counted in Cerenkov mode. self hoss The rapid phase of phorylation continues the experiment for an additional 10 minutes. Initial 10-8 at 15℃ The degree of phosphorylation completed at is 1/3 of the total self-phosphorylation signal, This probably reflects the phosphorylation of the cell site on the receptor. . Therefore, this 10-s interval was chosen to perform subsequent autophosphorylation experiments. Choose.

Self-phosphorylated ATP and RCF dependent EA to purified EG from A431 cells F-receptor (0.5 μg/assay) was incubated with EGF (0.85 μg/assay) for 20 min at 4°C. MM). The reaction was carried out at 15°C and Mg(Ac)t (60mM ), Tris-Mes buffer +, pH 7,6 (50rrM), [”PIA TP (without carrier, 5 μCi/assay) and increasing concentrations of non-number Jt [ Start by adding TP. This reaction was completed after 10-5 in SO3 sample buffer. Stop by addition. These samples were run on a 6% SDS polyacrylamide gel. make it move. The gel is dried and subjected to autoradiography as described above. Seki The consecutive radioactive band portions are cut out and counted in Cerenkov mode. in this way The amount of ATP measured was found to be 7.2 μM. l〇-S assay The protocol was used to measure the EGF concentration dependence of EGFRK autophosphorylation. Set.

Rat liver membranes were described by Cuatrecasas. prepared from six series of rat livers. Insulin purified with EA Septa are prepared according to the method of Zick et al. La purified by WGA In5RK (1,25 Mg) in human liver was combined with 330 nM insulin. in 50mM Tris-Mes buffer, pH 7.6, with or without Pre-incubate for 30 minutes at 22°C. This assay was performed at 22°C. , Mg(Ac)t (60mM), NaVO5 (40μM), [γ-”PIATP (125MM; 3-5μCi/assay) and poly(GT) at three concentrations It is started by adding a mixture containing [po1ba c+uiryr)]. Treatment with inhibitors When processing, the poly(GT) is always added at an appropriate concentration. for this assay The final concentration of insulin in the sample is 125 nM. The total volume of the assay is 4 It is 0 μm. After 20 minutes, transfer a 30 μl aliquot to 3-ply 7 Attman filter paper. applied on top and immersed in cold 10% TcA containing 0.01M sodium persulfate. Ru. - by drying the filter paper after washing at night and measuring Cerenkov radiation; Perform counting. In5RK-catalyzed phosphorylation of poly(GT) Michaelis -Follow maintenance kinetics.

did. After washing twice with ice-cold PBS, the cells were washed with lysis buffer (50- Hepes, p) 17.5.150mM Nacl, t, 5d MgClt, 1m EGTA, 10% glycerol, 1% Triton X-100, 1mM PMS F, 1 mg/ml aprotinin, 1 mg/ml oipeptin) 500μ m7 dishes were added and lysed by incubating for 5 minutes at 4°C. EG After F stimulation (500 Mg/ml; 10 min at 37°C), immunoprecipitation tests were performed with anti-E The self-phosphorylation reaction ( Aliquots of 50 μl, 3 μC1[γ-32P]ATP) into 2 or 10 MM Compounds were run for 2 minutes at 4°C in the presence of compounds. This reaction is performed on high-temperature electrophoresis samples. Stopped by addition of buffer. 5O3-PAGE analysis (7.5% els) 1, followed by autoradiography, and the reaction was recorded in the density of X-ray film. Quantified by tometric scanning. Testing the selective inhibitory properties of the compounds of the present invention Repeat the procedure using PDGF stimulation instead of EGF stimulation to Example IC,. (μM) The results of this assay indicate that the compounds of the invention inhibit PDGF receptor kinase. It has been shown that EGF receptor kinase is inhibited better than that of EGF receptor kinase.

Inhibition of cell proliferation measured by inhibition of DNA synthesis with human fibronectin (10M (by incubating at room temperature for 30 min at 10.5 ml/well). lXl0 per well in a 24-well Costar-CCO3tar) dish 'Cells were seeded at a density of 'cells. Grow these cells to confluence for 2 days. I set it. The medium was replaced with DMEM containing 0.5 bovine serum and grown for 36-48 hours. The cells were then treated with EGF (Toyobo, New York, NY; 20 ng/ml) or serum (10% bovine serum) and various concentrations of the inhibitory compound. Incubated with [1 H Cothymidine (NEN, Boston, MA)] After 6-24 hours, it was added at 0.5 μCi/ml and incubated for 2 hours. T The CA precipitated material was quantified using a scintillation counter.

cell culture Cells named HER14 and 721A (=DK) were cultured with endogenous EGF-levels. Septa-poor NIH3T3 cells (clone 2.2) (C, flyling (F) ryling), NCr, obtained from NIH) with poor tyrosine kinase activity. cDNA construct of new wild-type EGF-receptor or mutant EGF-receptor (Here, each Lys721 in the ATP-binding site is replaced with an Ala residue. prepared by transfection with (transformed). all cells 10 % bovine serum (Hyclone, Logan, Utah) ) grown inside.

The results obtained with the above experimental method demonstrate that the compounds within the scope of the present invention are useful proteins. Demonstrates tyrosine kinase inhibitory properties.

abstract For heteroarylethendiyl or heteroarylethenediyl aryl compounds, and the heteroaryl group is a monocyclic or bicyclic heteroaryl group, and the aryl group is a monocyclic or bicyclic heteroaryl group, and a monocyclic or bicyclic heteroaryl group or a bicyclic or tricyclic carboxylic group. iclic group, and the compound is optionally mono- or poly-substituted. , However, when the ethenediyl group has a geminal cyano substituent, the heteroatom lyl group is not a furyl or chenyl group), and pharmacological compositions containing the compound. and its use for inhibiting cell proliferation in patients suffering from such disorders.

international search report

Claims (31)

[Claims] 1. Heteroarylethendiyl or heteroarylethenediyl aryl compound or a pharmaceutically acceptable salt thereof to a patient suffering from a cell proliferation-related disorder. 2. A method for inhibiting cell proliferation in a patient, the method comprising: a cyclic or bicyclic heteroaryl group, and the aryl group is a monocyclic or bicyclic heteroaryl group; a heteroaryl group or a bicyclic or tricyclic carbocyclic group; The compound is optionally mono- or polysubstituted, with the proviso that the ethenediyl group has a geminal cyano substituent, the heteroaryl group is furyl or The above method, characterized in that the condition is that it is not a thienyl group. 2. a pharmacologically effective amount of a heteroaryl in admixture with a pharmaceutically acceptable carrier; Ethendiyl or heteroarylethenediyl aryl compounds or pharmaceutically acceptable Inhibits cell proliferation in patients suffering from cell proliferation-related disorders, including its salts that are administered. a pharmaceutical composition, wherein the heteroaryl group is a monocyclic or bicyclic heteroaryl group; a monocyclic or bicyclic heteroaryl group, and the aryl group is a monocyclic or bicyclic heteroaryl group or a bicyclic or tricyclic carbocyclic group, and the compound is optionally mono- or tricyclic. or poly-substituted, provided that the ethenediyl group carries a geminal cyano substituent. If so, the heteroaryl group is not a furyl or thienyl group. The above composition characterized in that: 3. Administration of the composition according to claim 2 to a patient suffering from a cell proliferation-related disorder A method of inhibiting cell proliferation in said patient. 4. Compounds represented by formula I below: R3-C(W)=CR2R1 I where W is an about 5 to about 7 membered monocyclic ring containing 1 or 2 N, O or S atoms. from about 8 to about 12 containing 1 or about 4 N, O or S atoms represents a heteroaryl ring system having two membered bicyclic rings; Optionally substituted with 1 to about 3 groups R4, R1 is alkyl, -CN, - CONRR, -CSNRR or -COOR, R is a hydrogen atom or an alkyl R2 represents -W, -CN, -CONHR5, -CON RR, -COOR, -CSNRR, -CH=C(CN)2, -C(NH2)=C (CN)2, ▲There are mathematical formulas, chemical formulas, tables, etc.▼, ▲There are mathematical formulas, chemical formulas, tables, etc.▼, ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ or about 9 to about 11 membered bicyclic arylcarboxyl ring systems or about 12 to about A 15-membered tricyclic arylcarboxyl ring system, which ring system optionally includes Optionally substituted with 1 to about 3 groups R4, where R3 is hydrogen, alkyl, -CN, -CH2CN, -CONRR, -CSNRR, -COOR or -CH=C(C N)CONH2, and each R4 independently represents alkyl, hydroxy, alkoxy, ha b, amino, mono- and dialkylamino, acetylamino, alkylthio, - CN, -CF3, nitro, -COOR, -CONRR, -CSNRR, -CHO , -CH=CHCOOH, -NHCO(CH2)2COOH or morpholino R5 is amino, -CONH2, ▲There are mathematical formulas, chemical formulas, tables, etc.▼ or ▲There are mathematical formulas, chemical formulas, tables, etc.▼; and each R6 is independently hydrogen, alkyl, hydroxy, alkoxy or halo. , R7 is independently alkyl, hydroxy, alkoxy, halo, nitro, carbo xy or carbalkoxy, n is 0 to about 6, and m is 1 to about 7. , However, when both R1 and R2 are -CN, W is a frill or a tie. provided that it is not nil. 5. A pharmacologically effective amount of the claimed item in admixture with a pharmaceutically acceptable carrier For cell proliferation-related disorders containing the compound described in item 4 or a pharmaceutically acceptable salt thereof. A pharmacological composition that inhibits cell proliferation in an affected patient. 6. Administering the composition according to claim 5 to a patient suffering from a cell proliferation-related disorder. A method of inhibiting cell proliferation in said patient. 7. In the above formula I, W contains 1 or 2 N, O or S atoms. or a 6-membered monocyclic aryl ring, or containing 1 to 4 N, O or S atoms represents a 9- to 10-membered bicyclic aryl ring, which ring optionally has 1 to about 3 may be substituted with R4 groups, R1 is -CN, -CONRR, -CSNR R or -COOR group, R represents hydrogen, alkyl or aralkyl, R 2 is -W, -CN, -CONHR5, -CONRR, -COOR, -CSNRR Or ▲There are mathematical formulas, chemical formulas, tables, etc.▼; , R3 is a hydrogen atom, and each R4 is independently alkyl, hydroxy , alkoxy or halo, R5 is amino, -CONH2, ▲ mathematical formula, chemical formula There are , tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼; represents, each R6 is independently hydrogen or an alkyl group, and each R7 is independently an alkyl group. , hydroxy, alkoxy, halo, nitro, carboxy or carbalkoxy The compound according to claim 4, wherein n is 0 to 4 and m is 2 to 6. . 8. A pharmacologically effective amount of the claimed item in admixture with a pharmaceutically acceptable carrier For cell proliferation-related disorders containing the compound described in item 7 or a pharmaceutically acceptable salt thereof. A pharmacological composition that inhibits cell proliferation in an affected patient. 9. Administering the composition according to claim 8 to a patient suffering from a cell proliferation-related disorder. A method of inhibiting cell proliferation in said patient. 10. In the above formula I, W is furyl, pyrrolyl, thienyl, thiazolyl, pyri Zyl, imidazolyl, isoimidazolyl, pyridazinyl, pyrimidinyl, benzo Furanyl, indolyl, indolinyl, indolinonyl, benzothienyl, ben Zothiazolyl, quinolinyl, isoquinolinyl, chromenyl, 1,3-benzodio The claimed compound is xoryl or 2,3-dihydro-1,4-benzodioxinyl. A compound according to scope item 7. 11. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; A cell proliferation-related disorder containing a compound according to item 10 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 12. The composition according to claim 11 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 13. In the above general formula I, W is pyridyl, indolyl, imidazolyl, ben Zothiazolyl, 1,3-benzodioxolyl or 2,3-dihydro-1,4- benzodioxynyl, and R5 is ▲There are mathematical formulas, chemical formulas, tables, etc.▼ or ▲There are mathematical formulas, chemical formulas, tables, etc.▼. The compound according to claim 10, which is 14. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; A cell proliferation-related disorder containing a compound according to item 13 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 15. The composition according to claim 14 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 16. In the above general formula I, each R4 is independently lower alkyl, hydroxy, lower alkoxy or halo, and each R7 is independently lower alkyl, hydroxy, alkoxy, halo, nitro, carboxy or carbalkoxy, as claimed A compound according to Range 13. 17. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; A cell proliferation-related disorder containing a compound according to item 16 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 18. The composition according to claim 17 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 19. The compound is 2-(4-pyridyl)-3-(9-phenanthryl)-2-propylene. 17. The compound according to claim 16, which is lopennitrile. 20. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; Cell proliferation-related disorders containing the compound according to item 19 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 21. The composition according to claim 20 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 22. The compound is 2-(3,4-dihydroxybenzoyl)-3-(5-indo 17. The compound according to claim 16, which is (lyl)-2-propenenitrile. 23. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; A cell proliferation-related disorder containing a compound according to item 22 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 24. The composition according to claim 23 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 25. The compound is 2-(3,4-dihydroxybenzoyl)-3-(3-indo 17. The compound according to claim 16, which is (lyl)-2-propenenitrile. 26. a claimed pharmaceutically effective amount in admixture with a pharmaceutically acceptable carrier; A cell proliferation-related disorder containing a compound according to item 25 or a pharmaceutically acceptable salt thereof A pharmacological composition that inhibits cell proliferation in a patient suffering from cancer. 27. The composition according to claim 26 is administered to patients suffering from cell proliferation-related disorders. A method of inhibiting cell proliferation in a patient comprising administering to the patient. 28. comprising administering a compound according to claim 2 to a patient suffering from psoriasis. A method of treating psoriasis in such a patient. 29. comprising administering a compound according to claim 5 to a patient suffering from psoriasis. A method of treating psoriasis in such a patient. 30. The compound according to claim 2 is administered to patients suffering from atherosclerosis. A method of treating atherosclerosis in a patient, the method comprising administering to a patient. 31. The compound according to claim 5 is administered to patients suffering from atherosclerosis. A method of treating atherosclerosis in a patient, the method comprising administering to a patient.