Diaryl hydantoin compound

Abstract

The present invention relates to diarylhydantoin compounds, including diarylthiohydantoin, as well as methods for their synthesis and use in the treatment of hormone refractory prostate cancer.

Description

[0001] The present invention is a divisional application of the Chinese invention application (filing date: March 29, 2006; application number: 202211664795.7; invention name: "Diarylhydantoin compound"). Technical Field

[0002] The present invention relates to diarylthiohydantoin compounds, methods for synthesizing them, and their use in treating hormone-refractory prostate cancer. This application claims priority to U.S. Provisional Application Serial Nos. 60 / 756,552, 60 / 750,351, and 60 / 680,835, which are incorporated herein by reference. Background of the Invention

[0003] Prostate cancer is the most common cancer in Western men and the second largest cause of cancer death. When cancer is confined to a local area, the disease can be cured by surgery or radiation. However, 30% of such cancers recur with distant metabolic diseases and other conditions that are diagnosed as serious illnesses. Serious illnesses are treated by castration and / or administration of antiandrogens, referred to as androgen deprivation therapy. Castration reduces the circulating levels of androgens and reduces the activity of androgen receptors (ARs). Administration of antiandrogens blocks AR function by competing for androgen binding, thereby reducing AR activity. Although initially effective, these treatments soon fail and cancer becomes hormone-resistant.

[0004] Recently, overexpression of AR has been identified and confirmed to be the cause of hormone-refractory prostate cancer. See Chen, CD, Welsbie, DS, Tran, C., Baek, SH, Chen, R., Vessella, R., Rosenfeld, MG, and Sawyers, CL, Molecular determinants of resistance to antiandrogen therapy, Nat. Med., 10: 33-39, 2004, which is incorporated herein by reference. Overexpression of AR is sufficient to develop from hormone sensitivity to hormone-refractory prostate cancer, meaning that better AR inhibitors than current drugs can slow the development of prostate cancer. It has been shown that AR and its bound ligands are essential for the growth of hormone-refractory prostate cancer, indicating that AR is still the target of the disease. It has also been shown that overexpression of AR converts anti-androgenic substances from antagonists to agonists (AR antagonists inhibit AR activity and AR agonists stimulate AR activity) in hormone-refractory prostate cancer. Data from this work explain why castration and antiandrogens fail to prevent the progression of prostate cancer and reveal an unrecognized nature of hormone-refractory prostate cancer.

[0005] Bicalutamide (trade name: Conside) is the most commonly used antiandrogen. Although it has an inhibitory effect on AR in hormone-sensitive prostate cancer, it cannot inhibit AR when cancer becomes hormone-resistant. Because it cannot prevent prostate cancer from developing from the hormone-sensitive stage to hormone-resistant disease, nor can it effectively treat hormone-refractory prostate cancer, two shortcomings of current antiandrogens are criticized. When AR is overexpressed in hormone-refractory prostate cancer, one is their weak antagonistic activity and the other is their strong agonistic activity. Therefore, there is a need for better AR inhibitors with more effective antagonistic activity and minimal agonistic activity to delay the development of the disease and to treat fatal hormone-refractory prostate cancer.

[0006] Because they are more selective and have fewer side effects, nonsteroidal antiandrogens, such as bicalutamide, are preferred over steroidal compounds for prostate cancer. Such compounds have been described in many patents, such as U.S. Patent No. 4,097,578, U.S. Patent No. 5,411,981, U.S. Patent No. 5,705,654, PCT International Application Nos. WO 97 / 00071 and WO 00 / 17163, and U.S. Published Patent Application No. 2004 / 0009969, all of which are incorporated herein by reference.

[0007] U.S. Patent No. 5,434,176 contains broad claims covering a large number of compounds, but only provides synthetic routes for a small number of these compounds, and only provides pharmacological data for two of them, and a person skilled in the art cannot easily foresee other specific compounds.

[0008] Since the mechanism of hormone-refractory prostate cancer is not known, there is no biological system described in these patents to test the effects of these compounds on hormone-refractory prostate cancer. In particular, the ability of AR overexpression in hormone-refractory prostate cancer to convert inhibitors from antagonists to agonists has not been recognized. Some new properties of hormone-refractory prostate cancer are reported in PCT applications US04 / 42221 and US05 / 05529, which are incorporated herein by reference. PCT International Application US05 / 05529 proposes a method for determining the androgen receptor antagonistic and agonistic properties of a compound. However, for each prepared compound, a time-consuming method for determining the antagonistic and agonistic properties of the compound must be determined. That is, there is no method for accurately predicting the properties of the relevant treatment of prostate cancer from the chemical structure of the compound alone.

[0009] There is a need for new thiohydantoin compounds with desirable pharmacological properties and synthetic routes to prepare them. Because activity is sensitive to even small structural changes, one compound may be effective in treating prostate cancer while a second compound may be ineffective, even if it differs from the first compound by only a small difference, such as the replacement of a single substituent.

[0010] The identification of compounds with high potency antagonistic androgen activity and minimal agonistic activity should overcome hormone-refractory prostate cancer (HRPC) and avoid or slow the development of hormone-sensitive prostate cancer (HSPC). Therefore, there is a need in the art to identify selective modulators of the androgen receptor, such as non-steroidal, non-toxic and tissue-selective modulators. SUMMARY OF THE INVENTION

[0011] The present invention provides a series of compounds having strong antagonistic activity and minimal agonistic activity against AR, and these compounds inhibit the growth of hormone-refractory prostate cancer.

[0012] The first aspect of the present invention relates to compounds having the following formula:

[0013]

[0014] wherein X is selected from trifluoromethyl and iodine,

[0015] wherein W is selected from O and NR5,

[0016] wherein R5 is selected from H, methyl and

[0017]

[0018] wherein D is S or O, and E is N or O, and G is alkyl, aryl, substituted alkyl, or substituted aryl; or

[0019] D is S or O and EG together are C1-C4 lower alkyl,

[0020] wherein R1 and R2 together comprise 8 or fewer carbon atoms and are selected from alkyl, substituted alkyl including haloalkyl, and cycloalkyl or substituted cycloalkyl together with the carbon to which they are attached,

[0021] wherein R is selected from the group consisting of hydrogen, halogen, methyl, C1-C4 alkoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methoxycarbonyl, acetylamino, methylsulfonylamino, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, and C1-C6 alkyl or alkenyl, optionally substituted with hydroxy, methoxycarbonyl, cyano, amino, amido, nitro, carbamoyl, or substituted carbamoyl including methylcarbamoyl, dimethylcarbamoyl and hydroxyethylcarbamoyl,

[0022] wherein R4 is selected from the group consisting of hydrogen, halogen, alkyl and haloalkyl,

[0023] Wherein R3 is not methylaminomethyl or dimethylaminomethyl.

[0024] Preferably in the first aspect, the compound wherein R5 is

[0025]

[0026] Preferably in the first aspect, the compound has the formula:

[0027]

[0028] wherein R3 is selected from hydroxy, methylcarbamoyl, methylcarbamoylpropyl, methylcarbamoylethyl, methylcarbamoylmethyl, methylsulfonylcarbamoylpropyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, carbamoylmethyl, carbamoylethyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoylpropyl, carboxypropyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, hydroxyethylcarbamoylethyl and hydroxyethoxycarbonylethyl, and

[0029] wherein R10 and R11 are both H, or are F and H, or H and F, respectively.

[0030] Preferably, wherein R10 and R11 are both H. Preferably, wherein R10 and R11 are F and H, respectively. Preferably, wherein R3 is methylcarbamoyl. Preferably, wherein R3 is methylcarbamoyl and R10 and R11 are F and H, respectively.

[0031] Preferably, in the first aspect, the compound, wherein R1 and R2 are independently methyl, or together with the carbon to which they are attached, are cycloalkyl having 4-5 carbon atoms, and R3 is selected from carbamoyl, alkylcarbamoyl, carbamoylalkyl and alkylcarbamoylalkyl, and R4 is H or F. Preferably, wherein R4 is 3-fluoro.

[0032] Preferably, in the first aspect, the compound, wherein R1 and R2 are independently methyl, or together with the carbon atoms to which they are attached, are cycloalkyl groups of 4-5 carbon atoms,

[0033] R3 is selected from cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-(1,1-dimethylethoxycarbonyl)-1-piperazinyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxyethoxycarbonyl-substituted alkyl and 3-cyano-4-trifluoromethylphenylcarbamoyl, and R4 is F.

[0034] Preferably in the first aspect, the compound has the formula:

[0035]

[0036] wherein R3 is selected from methylcarbonyl, methoxycarbonyl, acetylamino and methylsulfonylamino, and R4 is selected from F and H.

[0037] Preferably in the first aspect, the compound has the formula:

[0038]

[0039] wherein R4 is selected from F and H.

[0040] Preferably in the first aspect, the compound wherein R1 and R2 together with the carbon to which they are attached are:

[0041]

[0042] Preferably, in the first aspect, the compound is selected from compounds of Class 1 and Class 2. Preferably, in the first aspect, the compound has the following formula:

[0043]

[0044] Preferably in the first aspect, the compound has the formula:

[0045]

[0046] Preferably in the first aspect, the compound has the formula:

[0047]

[0048] Preferably in the first aspect, the compound has the formula:

[0049]

[0050] Preferably in the first aspect, the compound has the formula:

[0051]

[0052] The second aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound according to the first aspect and each preferred aspect or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0053] A third aspect of the present invention relates to a method for treating a hyperproliferative disease, comprising administering the pharmaceutical composition of the second aspect of the present invention to a patient in need of such treatment, thereby treating the hyperproliferative disease. Preferably, the composition is administered at a dose of the compound ranging from about 0.001 mg per kilogram of body weight per day to about 100 mg per kilogram of body weight per day. Preferably, the composition is administered at a dose of the compound ranging from about 0.01 mg per kilogram of body weight per day to about 100 mg per kilogram of body weight per day. Preferably, the composition is administered at a dose of the compound ranging from about 0.1 mg per kilogram of body weight per day to about 10 mg per kilogram of body weight per day. Further preferably, the composition is administered at a dose of about 1 mg per kilogram of body weight per day. Preferably, the hyperproliferative disease is hormone-refractory prostate cancer. Preferably, the compound is administered intravenously, by injection into a tissue, intraperitoneally, orally, or intranasally. Preferably, the composition has a form selected from the group consisting of a solution, a dispersion, a suspension, a powder, a capsule, a tablet, a pill, a time-release capsule, a time-release tablet and a time-release pill.

[0054] A fourth aspect of the present invention relates to a method for treating a hyperproliferative disease, comprising administering the composition of the second aspect of the present invention to a patient in need of such treatment, thereby treating the hyperproliferative disease. Preferably, the composition is administered at a dose ranging from about 0.1 mg per kilogram of body weight per day to about 10 mg per kilogram of body weight per day. Preferably, the composition is administered at a dose of about 1 mg per kilogram of body weight per day. Preferably, the composition is administered orally. Preferably, the composition has a form selected from the group consisting of: capsules, tablets, and pills. Preferably, the compound is selected from RD162', RD162", RD169, or RD170, or a pharmaceutically acceptable salt thereof. Preferably, the compound is N-methyl-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-2-fluorobenzamide [RD162] or a pharmaceutically acceptable salt thereof.

[0055] A fifth aspect of the present invention relates to a method for synthesizing a diaryl compound of the following formula:

[0056]

[0057] The method comprises mixing compound I and compound II in a first polar solvent to form a mixture;

[0058]

[0059] Heat the mixture,

[0060] A second polar solvent, which may be the same as or different from the first polar solvent, and an aqueous solution of an acid are added to the mixture,

[0061] The mixture was refluxed,

[0062] The mixture is cooled and mixed with water, and

[0063] separating the diaryl compound from the mixture,

[0064] wherein R51 comprises an alkyl chain of 1 to 4 carbon atoms, R52 is selected from the group consisting of cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl and hydroxyethoxycarbonyl-substituted alkyl, and R53 is selected from the group consisting of F and H. Preferably, wherein R51 comprises an alkyl chain of 1 to 2 carbon atoms, R52 is selected from the group consisting of carbamoyl and methylcarbamoyl, and R53 is F.

[0065] A sixth aspect of the present invention relates to a method for synthesizing the compound of the following formula:

[0066]

[0067] The method comprises

[0068] mixing 4-isothiocyanato-2-trifluoromethylbenzonitrile and N-methyl-4-(1-cyanocyclobutylamino)-2-fluorobenzamide in dimethylformamide to form a first mixture,

[0069] heating the first mixture to form a second mixture,

[0070] To this second mixture is added an alcohol and an acid to form a third mixture,

[0071] refluxing the third mixture to form a fourth mixture,

[0072] cooling the fourth mixture,

[0073] The fourth mixture was mixed with water and the organic layer was extracted,

[0074] The compound was isolated from the organic layer.

[0075] A seventh aspect of the present invention relates to a method for preparing compound [RD162'], said method comprising

[0076] combining N-methyl-2-fluoro-4-(1,1-dimethyl-cyanomethyl)-aminobenzamide and 4-isothiocyanato-2-trifluoromethylbenzonitrile in DMF and heating to form a first mixture;

[0077] adding an alcohol and an acid to this first mixture to form a second mixture;

[0078] reflux the second mixture;

[0079] Cooling the second mixture,

[0080] mixing the second mixture with water and extracting the organic layer;

[0081] From the organic layer, the mixture was separated.

[0082] An eighth aspect of the present invention relates to a method for preparing compound [RD162"], said method comprising

[0083] mixing N-methyl-2-fluoro-4-(1-cyanocyclopentyl)aminobenzamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile, and DMF and heating to reflux to form a first mixture;

[0084] adding an alcohol and an acid to the first mixture to form a second mixture;

[0085] reflux the second mixture;

[0086] Cooling the second mixture;

[0087] The second mixture is mixed with water and the organic layer is extracted;

[0088] The compound was isolated from the organic layer.

[0089] A ninth aspect of the present invention relates to a method for preparing compound [RD169], said method comprising

[0090] mixing N,N-dimethyl-4-[4-(1-cyanocyclobutylamino)phenyl]butanamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile, and DMF and heating to reflux to form a first mixture;

[0091] adding an alcohol and an acid to this first mixture to form a second mixture;

[0092] reflux the second mixture;

[0093] Cooling the second mixture;

[0094] The second mixture is mixed with water and the organic layer is extracted;

[0095] The compound was isolated from the organic layer.

[0096] A tenth aspect of the present invention relates to a method for preparing compound [RD170], said method comprising

[0097] Mix DMSO, dichloromethane and oxalyl chloride to form a first mixture,

[0098] To this first mixture was added 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanamide to form a second mixture;

[0099] adding triethylamine to the second mixture to form a third mixture;

[0100] The third mixture was warmed and quenched with aqueous NH4Cl to form a fourth mixture;

[0101] extracting an organic layer from the fourth mixture;

[0102] The compound was isolated from the organic layer.

[0103] The eleventh aspect of the present invention relates to compounds of the following formula:

[0104]

[0105] Where R5 is CN or NO2 or SO2R 11 ,

[0106] wherein R6 is CF3, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, halogen,

[0107] Wherein A is sulfur (S) or oxygen (O),

[0108] Wherein B is O or S or NR8,

[0109] wherein R8 is selected from H, methyl, aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, SO2R 11 NR 11 R 12 、(CO)OR 11 、(CO)NR 11 R 12 、(CO)R 11 、(CS)R 11 、(CS)NR 11 R 12 、(CS)OR 11 、

[0110]

[0111] wherein D is S or O, and E is N or O, and G is alkyl, aryl, substituted alkyl, or substituted aryl; or

[0112] D is S or O and EG together are C1-C4 lower alkyl,

[0113] wherein R1 and R2 are independently alkyl, haloalkyl, hydrogen, aryl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic heterocyclic group or non-aromatic heterocyclic group, substituted aromatic heterocyclic group or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, or R1 and R2 are linked to form a ring, which can be a heterocyclic group, a substituted heterocyclic group, a cycloalkyl, a substituted cycloalkyl,

[0114]

[0115] wherein X is carbon or nitrogen and may be at any position on the ring, and

[0116] wherein R3, R4 and R7 are independently selected from hydrogen, halogen, methyl, methoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methylcarbamoyl-substituted alkyl, dimethylcarbamoyl-substituted alkyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxy-substituted alkyl, hydroxy-substituted alkenyl, carbamoyl-substituted alkenyl, methoxycarbonyl-substituted alkyl, cyano-substituted alkyl, Aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, SO2R 11 NR 11 R 12 NR 12 (CO)OR 11 、NH(CO)NR 11 R 12 NR 12 (CO)R 11 、O(CO)R 11 、O(CO)OR 11 、O(CS)R 11 NR 12 (CS)R 11 、NH(CS)NR 11 R 12 NR 12 (CS)OR 11, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, haloalkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-(1,1-dimethylethoxy)carbonyl)-1-piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxyethoxycarbonyl-substituted alkyl, 3-cyano-4-trifluoromethylphenylcarbamoyl,

[0117] where R 11 and R 12 are independently hydrogen, aryl, aralkyl, substituted aralkyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, or substituted cycloalkyl, or R 11 and R 12 They may be linked to form a ring, which may be an aromatic heterocyclic group or a non-aromatic heterocyclic group, a substituted aromatic heterocyclic group, a cycloalkyl group, or a substituted cycloalkyl group.

[0118] Preferably, the compound has significant androgen receptor antagonist activity and substantially no agonist activity against hormone-refractory prostate cancer cells.

[0119] A twelfth aspect of the present invention relates to a method comprising:

[0120] providing at least one compound according to the eleventh aspect;

[0121] measuring inhibition of androgen receptor activity by the compound and determining whether the inhibition is above a first predetermined level,

[0122] measuring stimulation of androgen receptor activity in hormone-resistant cancer cells by the compound and determining whether the stimulation is below a second predetermined level,

[0123] If the inhibition is above a first predetermined level and the stimulation is below a second predetermined level, the compound is selected.

[0124] Preferably, wherein said predetermined levels are those of bicalutamide.

[0125] Preferably, the step of measuring inhibition comprises measuring inhibitory concentration (IC50) in an AR-responsive reporter system or a prostate-specific antigen secretion system.

[0126] Preferably, the step of measuring stimulation comprises measuring the fold induction by increasing the concentration in an AR response reporter system or a prostate specific antigen secretion system.

[0127] Preferably, said step of measuring inhibition and / or stimulation comprises measuring the effect of said compound on tumor growth in an animal. Detailed Description of the Invention

[0128] The present invention includes compounds having the formula:

[0129]

[0130] wherein X is selected from trifluoromethyl and iodine, wherein W is selected from O and NR5, wherein R5 is selected from the following groups: H, methyl and

[0131]

[0132] wherein D is S or O, and E is N or O, and G is alkyl, aryl, substituted alkyl or substituted aryl; or D is S or O and EG together are C1-C4 lower alkyl,

[0133] wherein R1 and R2 together contain 8 or less carbon atoms and are selected from the group consisting of alkyl, substituted alkyl including haloalkyl, and, together with the carbon atom to which they are attached, cycloalkyl or substituted cycloalkyl,

[0134] wherein R3 is selected from the group consisting of hydrogen, halogen, methyl, C1-C4 alkoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methoxycarbonyl, acetylamino, methylsulfonylamino, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, and C1-C6 alkyl or alkenyl, optionally substituted with hydroxy, methoxycarbonyl, cyano, amino, amido, nitro, carbamoyl, or substituted carbamoyl including methylcarbamoyl, dimethylcarbamoyl and hydroxyethylcarbamoyl,

[0135] wherein R4 is selected from the group consisting of hydrogen, halogen, alkyl and haloalkyl, and

[0136] Wherein R3 is not methylaminomethyl or dimethylaminomethyl.

[0137] R5 can be

[0138]

[0139] The compound may have the following structure:

[0140]

[0141] wherein R3 is selected from the group consisting of hydroxy, methylcarbamoyl, methylcarbamoylpropyl, methylcarbamoylethyl, methylcarbamoylmethyl, methylsulfonylcarbamoylpropyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, carbamoylmethyl, carbamoylethyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoylpropyl, carboxypropyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, hydroxyethylcarbamoylethyl, and hydroxyethoxycarbonylethyl, and

[0142] wherein R10 and R11 are both H, or are respectively F and H, or H and F. In some embodiments, R10 and R11 may both be H, or are respectively F and H. R3 may be methylcarbamoyl.

[0143] In some embodiments, R1 and R2 are independently methyl, or a cycloalkyl group of 4 to 5 carbon atoms together with the carbon atom to which they are attached, and R3 is selected from the group consisting of carbamoyl, alkylcarbamoyl, carbamoylalkyl, and alkylcarbamoylalkyl, and R4 is H or F or R4 is 3-fluoro.

[0144] In other embodiments, R1 and R2 are independently methyl, or cycloalkyl of 4 to 5 carbon atoms, taken together with the carbon atom to which they are attached, R3 is selected from the group consisting of cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, acetamido, methylsulfonylamino, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-(1,1-dimethylethoxycarbonyl)-1-piperazinyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxyethoxycarbonyl-substituted alkyl, and 3-cyano-4-trifluoromethylphenylcarbamoyl, and R4 is F.

[0145] The compounds of the present invention may have the following structure:

[0146]

[0147] wherein R3 is selected from the group consisting of methylcarbonyl, methoxycarbonyl, acetylamino and methanesulfonylamino, and R4 is selected from F and H.

[0148] The compounds of the present invention may have the following structure:

[0149]

[0150] wherein R4 is selected from F and H.

[0151] In an embodiment of the present invention, wherein R1 and R2 together with the carbon atom to which they are attached are

[0152]

[0153] The compounds of the present invention may be those listed in Class 1, Class 2, Class 3 and / or Class 4 below. Specific compounds of the present invention include

[0154]

[0155]

[0156] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any of the foregoing compounds or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent.

[0157] The present invention includes methods for treating hyperproliferative diseases comprising administering such pharmaceutical compositions to a patient in need thereof to treat a hyperproliferative condition. The hyperproliferative condition can be hormone-refractory prostate cancer. Dosages can range from about 0.001 mg / kg body weight per day to about 100 mg / kg body weight per day, about 0.01 mg / kg body weight per day to about 100 mg / kg body weight per day, about 0.1 mg / kg body weight per day to about 10 mg / kg body weight per day, or about 1 mg / kg body weight per day.

[0158] The compound can be administered intravenously, by injection into a tissue, intraperitoneally, orally or intranasally. The composition can have a form selected from the group consisting of solutions, dispersions, suspensions, powders, capsules, tablets, pills, time-release capsules, time-release tablets and time-release pills.

[0159] The compound to be administered may be selected from the following: RD162', RD162", RD169 or RD170, or a pharmaceutically acceptable salt thereof. The compound to be administered may be RD162 or a pharmaceutically acceptable salt thereof.

[0160] The present invention provides a method for synthesizing a diaryl compound of the following formula:

[0161]

[0162] It comprises compound I

[0163]

[0164] With compound II

[0165]

[0166] The method comprises the steps of: mixing in a first polar solvent to form a mixture, heating the mixture, adding a second polar solvent, which is the same as or different from the first solvent, and an aqueous acid solution to the mixture, refluxing the mixture, cooling the mixture and mixing with water, and isolating from the mixture a diaryl compound wherein R51 comprises an alkyl chain of 1 to 4 carbon atoms, and R52 is a group selected from the group consisting of cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, Methylaminomethyl, dimethylaminomethyl, methanesulfonyloxymethyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methanesulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-methanesulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl and hydroxyethoxycarbonyl-substituted alkyl, and R53 is selected from F and H.

[0167] R51 may include an alkyl chain of 1-2 carbon atoms, R52 may be selected from carbamoyl and methylcarbamoyl, and R53 may be F.

[0168] The present invention provides a method for synthesizing the compound of the following formula:

[0169]

[0170] The method comprises mixing 4-isothiocyanato-2-trifluoromethylbenzonitrile and N-methyl-4-(1-cyanocyclobutylamino)-2-fluorobenzamide in dimethylformamide to form a first mixture, heating the first mixture to form a second mixture, adding an alcohol and an acid to the second mixture to form a third mixture, refluxing the third mixture to form a fourth mixture, cooling the fourth mixture, mixing the fourth mixture with water, extracting an organic layer, and separating the compound from the organic layer.

[0171] Likewise, the present invention provides a method for synthesizing RD162', comprising mixing N-methyl-2-fluoro-4-(1,1-dimethyl-cyanomethyl)-aminobenzamide and 4-isothiocyanato-2-trifluoromethylbenzonitrile in DMF and heating to form a first mixture, and performing the above treatment.

[0172] The present invention also provides a method for synthesizing RD162", which comprises mixing N-methyl-2-fluoro-4-(1-cyanocyclopentyl)aminobenzamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile and DMF, heating to reflux to form a first mixture, and performing the above treatment.

[0173] The present invention further provides a method for synthesizing RD169, which comprises mixing N,N-dimethyl-4-[4-(1-cyanocyclobutylamino)phenyl]butanamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile and DMF and heating to reflux to form a first mixture, and performing the above treatment.

[0174] The present invention provides a method for synthesizing RD170, which includes mixing DMSO, dichloromethane and oxalyl chloride to form a first mixture, adding 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanamide to the first mixture to form a second mixture, adding triethylamine to the second mixture to form a third mixture, warming the third mixture and quenching it with an NH4Cl aqueous solution to form a fourth mixture, extracting an organic layer from the fourth mixture, and isolating a compound from the organic layer.

[0175] Other compounds according to the present invention have the following formula:

[0176]

[0177] wherein R5 is CN or NO2 or SO2R11, wherein R6 is CF3, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, halogen, wherein A is sulfur (S) or oxygen (O), wherein B is O or S or NR8, wherein R8 is selected from the group consisting of H, methyl, aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, SO2R11, NR11R12, (CO)OR11, (CO)NR11R12, (CO)R11, (CS)R11, (CS)NR11R12, (CS)OR11,

[0178]

[0179] wherein D is S or O and E is N or O, and G is alkyl, aryl, substituted alkyl or substituted aryl; or D is S or O and EG together are C1-C4 lower alkyl,

[0180] wherein R1 and R2 are independently alkyl, haloalkyl, hydrogen, aryl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic heterocyclic group or non-aromatic heterocyclic group, substituted aromatic heterocyclic group or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, or R1 and R2 are linked to form a ring, and the ring can be heterocyclic group, substituted heterocyclic group, cycloalkyl, substituted cycloalkyl,

[0181]

[0182] wherein X is carbon or nitrogen and may be located anywhere in the ring, and

[0183] wherein R3, R4 and R7 are independently selected from the group consisting of hydrogen, halogen, methyl, methoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methylcarbamoyl-substituted alkyl, dimethylcarbamoyl-substituted alkyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxy-substituted alkyl, hydroxy-substituted alkenyl, carbamoyl-substituted alkenyl, methoxycarbonyl-substituted alkyl, cyano-substituted alkyl, Aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, SO2R11, NR11R12, NR12(CO)OR11, NH(CO)NR11R12, NR12(CO)R11, O(CO)R11, O(CO)OR11, O(CS)R11, NR12(CS)R11, NH(CS)NR11R12, NR12(CS)OR11, arylalkyl, arylalkenyl, arylalkynyl, aromatic heterocyclic group or non-aromatic heterocyclic group, substituted aromatic heterocyclic group or non-aromatic a heterocyclic group, a cycloalkyl group, a substituted cycloalkyl group, a haloalkyl group, a methylsulfonylcarbamoyl-substituted alkyl group, a methylaminomethyl group, a dimethylaminomethyl group, a methylsulfonyloxymethyl group, a methoxycarbonyl group, an acetylamino group, a methylsulfonylamino group, a carbamoyl-substituted alkyl group, a carboxymethyl group, a methoxycarbonylmethyl group, a methylsulfonyl group, a 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl group, a carboxy-substituted alkyl group, a 4-(1,1-dimethylethoxy)carbonyl)-1-piperazinyl group, a hydroxyethylcarbamoyl-substituted alkyl group, a hydroxyethoxycarbonyl-substituted alkyl group, a 3-cyano-4-trifluoromethylphenylcarbamoyl group,

[0184] wherein R11 and R12 are independently hydrogen, aryl, aralkyl, substituted aralkyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, or substituted cycloalkyl, or R11 and R12 may be linked to form a ring which may be aromatic or non-aromatic heterocyclic group, substituted aromatic heterocyclic group, cycloalkyl, or substituted cycloalkyl.

[0185] These compounds have significant androgen receptor antagonist activity and essentially no agonist activity.

[0186] The present invention includes a method comprising providing at least one such compound, testing the compound for inhibition of androgen receptor activity, and determining whether the inhibition is above a first predetermined level, testing the compound for stimulation of androgen receptor activity in hormone-resistant cancer cells, and determining whether the stimulation is below a second predetermined level, and selecting the compound if the inhibition is above a first predetermined level and the stimulation is below a second predetermined level. The predetermined levels may be those of bicalutamide. The measuring inhibition step may include measuring inhibitory concentrations (IC50) in an AR-responsive reporter system or a prostate-specific antigen secretion system. The measuring stimulation step may include measuring fold induction by increasing the concentration in an AR-responsive reporter system or a prostate-specific antigen secretion system. Measuring inhibition and / or stimulation methods may include measuring the effect of the compound on tumor growth in animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0187] The following figures show the results of pharmacological testing of some compounds.

[0188] Figure 1 Figure 1 depicts a graph showing the agonistic effect of bicalutamide on LNCaP-AR. Bicalutamide agonist activity in AR-overexpressed hormone-refractory prostate cancer. LNCaP cells with overexpressed AR were treated with increasing concentrations of DMSO (as a vehicle) and bicalutamide in the absence of R1881. The activity of the AR response reporter was measured.

[0189] Figure 2 Figure 1 depicts a test of bicalutamide's antagonistic activity against LNCaP-AR. Agonistic activity of bicalutamide in hormone-sensitive prostate cancer. LNCaP cells were treated with increasing concentrations of DMSO (as vehicle) and bicalutamide in the absence of R1881. The activity of the AR-responsive reporter was measured.

[0190] Figure 3Figure 2 is a graph depicting the effects of compounds on LNCaP-AR.

[0191] Figure 4 Figure 2 is a graph depicting the effect of compound LNCaP-AR.

[0192] Figure 5 Graph depicting the inhibitory effect on LNCaP-AR.

[0193] exist Figure 6-10 Among them, Example 5-3b is RD7 and Example 7-3b is RD37.

[0194] Figure 6 Inhibition of growth of AR-overexpressing LNCaP cells. Androgen-deficient LNCaP cells with overexpressed AR were treated with increasing concentrations of DMSO (as vehicle) and test substances in the presence of 1 pM R1881. After 4 days of culture, cell growth was measured by MTS assay.

[0195] Figure 7. Inhibitory effect on the growth of an AR-overexpressing LNCaP xenograft model. Mice with established LN-AR xenograft tumors were randomized and treated orally once daily with the indicated compounds. Tumor size was measured using a scale. (A) Mice were treated with 1 mg / kg body weight of bicalutamide, Example 7-3b, or vehicle for 44 days. (B) Mice were treated with vehicle, 0.1, 1, or 10 mg / kg body weight of Example 7-3b for 44 days.

[0196] Figure 8 Inhibitory effect on PSA expression in an AR-overexpressing LNCaP xenograft model. Mice were treated orally with vehicle, 0.1, 1, or 10 mg of Example 7-3b per kilogram of body weight once daily for 44 days. After 44 days of treatment, tumors were removed from the mice, tumor lysates were extracted, and PSA levels in the tissue lysates were measured by ELISA.

[0197] Figure 9. Inhibitory effects on growth and PSA in a hormone-resistant LAPC4 xenograft model. Mice with established tumors were randomized and treated orally with 1 mg / kg body weight of bicalutamide, Example 7-3b, or vehicle once daily for 17 days. (A) Tumor size was measured using a cytometer. (B) After 17 days of treatment, tumors were removed from mice, tumor lysates were extracted, and PSA levels in the tissue lysates were measured using ELISA.

[0198] Figure 10 Inhibitory effect on the growth of hormone-sensitive prostate cancer cells. Androgen-deficient LNCaP cells were treated with increasing concentrations of DMSO (as vehicle) and test compounds in the presence of 1 pM R1881. After 4 days of culture, cell growth was measured by MTS assay.

[0199] Figure 11 Figure 2 shows tumor size. AR-overexpressing LNCaP cells were subcutaneously injected into the flanks of castrated SCID mice. When tumors reached approximately 100 cubic millimeters, they were randomly divided into five groups of nine animals each. Once these tumors reached this volume, they were orally administered with vehicle, bicalutamide, or RD162 at 10 or 50 mg / kg daily. Tumors were measured in three dimensions using a 3D scale: width, length, and height.

[0200] Figure 12 describes the experimental results of tumor size. At day 18, 3 hours after the last dose treatment, animals were imaged by an optical CCD camera. In order to test luciferase activity with photons / second (photon / second), ROIs were peeled off from the tumor. The right column is a representative ROI measurement.

[0201] Figure 13 Graph depicting the pharmacokinetic profiles of RD162 following intravenous (upper curve) and oral administration (lower curve).

[0202] Figure 14 Figure 2 depicts the uptake of PSA in LN-AR cells measured after treatment with various doses of several compounds.

[0203] Figure 15 is a table providing some characteristics of the compounds. Figure 15 is also a graph of the pharmacokinetic characteristics of some compounds presented as a function of compound serum concentration versus time.

[0204] Figure 16 Figure 1 is a graph depicting prostate weight after treatment with various compounds. 10, 25, or 50 mg of compound were administered daily per kilogram of body weight, as indicated by the markings in the columns. The compounds were administered to healthy FVB mice. After 14 days of compound treatment, genitourinary tract weight was measured by removing and weighing the semi-vesicles, prostate, and bladder. 3 mice were administered the specified compound to obtain the data shown in the columns in the figure. One group of mice was not treated with the compound: the data shown are indicated by the "untreated" marking in the columns. Another group of mice was treated only with vehicle solution: the data shown are indicated by the "vehicle" marking in the columns.

[0205] Figure 17 To show the Figure 6 Figure 4 shows a diagram of a PSA test performed according to the experimental protocol in FIG.

[0206] Figure 18 is a graph showing the effect of various dose regimens of RD162 on tumor volume.

[0207] FIG. 19 is a graph showing the ratio of photon emission for 17 days relative to day 0 after treatment with 0.1, 1, and 10 mg / kg body weight per day of RD162 and without RD162 treatment.

[0208] Figure 20 shows the experimental results of LN-AR (HR) cell line injected into SCID mice to induce tumor growth. One group of mice was treated with compound RD162 at a dosage of 10 mg per kg body weight per day, and another group of mice was treated with vehicle solution only. (A) The relative tumor volume and time function shown for each group of mice. (B) The images of the photon emission associated with luciferase in each group of mice at 31 days were displayed as colored profiles. (C) The ratio of the photon emission associated with luciferase activity shown for some time periods of each group of mice.

[0209] FIG21 is a graph showing PSA uptake associated with LN-AR cells treated with various concentrations of RD162, RD162′, RD162″, and RD170 and vehicle solution.

[0210] Figure 22 Figure 2 shows PSA uptake associated with LN-CaP cells treated with various concentrations of RD37, RD131, RD162, bicalutamide, and DMSO.

[0211] Figure 23 shows the experimental results of wild-type non-transgenic mice (WT), castrated luciferase transgenic mice (Cast) and non-castrated luciferase transgenic mice (Intact). The data show that the luciferase transgenic mice produced with 12.5mg per kg body weight of implanted testosterone pellets have a 90-day release cycle (T / Cast) and the data show that the luciferase transgenic mice produced with 12.5mg per kg body weight of implanted testosterone pellets have a 90-day release cycle (Intact+T) are not-castrated. The data show that the luciferase transgenic mice processed with implanted testosterone pellets and bicalutamide (BIC+T / Cast) or RD162 (RD162+T / Cast) of 10mg per kg body weight every day are castrated. (A) genitourinary tract weight at 14 days. (B) photon emission rate at 14 days. In all cases, no disease state was induced in which hormones were ineffective.

[0212] Figure 24 The graph shows the luciferase activity of L1AR cell line after administration of various compounds at concentrations ranging from 125 nmol to 1000 nmol.

[0213] Figure 25The graph shows the luciferase activity of LN / AR cell lines after administration of various compounds at concentrations ranging from 1.25 to 10 μmol.

[0214] Figure 26 The graph shows the luciferase activity of 4AR cell lines after administration of various compounds at concentrations ranging from 1.25 to 10 μmol.

[0215] Figure 27 The graph shows the PSA levels of 1AR cell line after administration of various compounds at concentrations ranging from 1.25 to 10 μmol.

[0216] Figure 28 The graph shows the PSA levels of LN / AR cell lines after administration of various compounds at concentrations ranging from 125 nmol to 1000 nmol.

[0217] Figure 29 The graph shows the luciferase activity of various compounds administered at concentrations ranging from 125 nmol to 1000 nmol. Detailed Description of the Invention

[0218] Embodiments of the present invention are discussed in detail below. In describing the embodiments, specific terminology is used for clarity. However, the present invention is not limited by the terminology so selected. Those skilled in the relevant art will recognize that other equivalents can be used, and other methods can be developed without departing from the spirit and scope of the present invention. All incorporated references are incorporated herein by reference as if each had been individually cited.

[0219] Synthesis of Diarylhydantoin Compounds

[0220] The present invention provides a method for synthesizing diarylthiohydantoins having the following formula:

[0221]

[0222] Wherein R71 comprises an alkyl chain of 1-4 carbon atoms. For example, R72 can be a carbamoyl group such as -(CO)NH2, or a methylcarbamoyl group such as -(CO)NHCH3. An amide group in which the carbon atom of the carbonyl group is attached to another structure is referred to as a carbamoyl substituent. For example, R73 can be a fluorine atom or a hydrogen atom. That is, the fluorine atom can be attached to any carbon atom or nitrogen atom on the right aromatic ring that is not attached to the R72 substituent. Alternatively, no fluorine atom can be attached to a carbon atom or nitrogen atom on the right aromatic ring that is not attached to the R72 substituent. For example, a hydrogen atom can be attached to every carbon atom or nitrogen atom on the right aromatic ring that is not attached to the R72 substituent.

[0223] For example, as further shown below (see, e.g., Figure 3 、5 , 11-13), a compound having the formula

[0224]

[0225] It showed surprisingly potent antagonistic activity and minimal agonistic activity against AR overexpressed in hormone-refractory prostate cancer.

[0226] A list of some compounds according to the present invention is listed in Tables 5-11. These compounds are divided into several categories, with compounds in categories 1 to 3 expected to be superior to bicalutamide in treating prostate cancer, compounds in category 4 being comparable in effectiveness to bicalutamide, and compounds in categories 5 and 6 being inferior to bicalutamide in treating prostate cancer. The scheme for arranging the compounds into categories is described in more detail below.

[0227] definition

[0228] As used herein, the term "alkyl" refers to a branched or unbranched hydrocarbon chain, preferably having 1 to 8 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, 2-methylpentylpentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, etc. "Substituted alkyl" includes alkyl groups optionally substituted with one or more functional groups attached to the chain, such as hydroxy, bromo, fluoro, chloro, iodo, mercapto or thio, cyano, alkylthio, heterocyclyl, aryl, heteroaryl, carboxyl, carbalkoyl, alkyl, alkenyl, nitro, amino, alkoxy, amido, and the like to form alkyl groups such as trifluoromethyl, 3-hydroxyhexyl, 2-carboxypropyl, 2-fluoroethyl, carboxymethyl, cyanobutyl, and the like.

[0229] Unless otherwise indicated, the term "cycloalkyl" as used herein alone or as part of another group includes saturated or partially saturated (containing one or more double bonds) cyclic hydrocarbon groups containing 1 to 3 rings, including monocyclic alkyl, bicyclic alkyl and tricyclic alkyl groups, containing a total of 3 to 20 carbon atoms forming the ring, preferably 3 to 10 carbon atoms forming the ring, and which may be fused to 1 or 2 aromatic rings as described for aryl, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl. "Substituted cycloalkyl" includes cycloalkyl groups optionally substituted with one or more substituents such as halogen, alkyl, alkoxy, hydroxy, aryl, aryloxy, arylalkyl, cycloalkyl, alkylamido, alkylnolyamino, oxo, acetyl, arylcarbonylamino, amino, nitro, cyano, thiol and / or alkylthio and / or any substituent included in the definition of "substituted alkyl" above. For example, etc.

[0230] As used herein, unless otherwise indicated, the term "alkenyl", used alone or as part of another group, refers to a straight or branched chain group having 2 to 20 carbons, preferably 2 to 12 carbons, and more preferably 2 to 8 carbons in the backbone, which includes one or more double bonds in the backbone, such as ethenyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 3-octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl, 4-dodecenyl, 4,8,12-tetradecatriene, etc. "Substituted alkenyl" includes alkenyl groups optionally substituted with one or more substituents, such as those included in the above definitions of "substituted alkyl" and "substituted cycloalkyl".

[0231] Unless otherwise indicated, the term "alkynyl", used alone or as part of another group, refers to a straight or branched chain group having 2 to 20 carbons, preferably 2 to 12 carbons, and more preferably 2 to 8 carbons in the backbone, including one or more triple bonds in the backbone, such as 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, 4-decynyl, 3-undecynyl, 4-dodecynyl, etc. "Substituted alkynyl" includes alkynyl groups optionally substituted with one or more substituents, such as the substituents included in the above definitions of "substituted alkyl" and "substituted cycloalkyl".

[0232] The terms "arylalkyl," "arylalkenyl," and "arylalkynyl," used alone or as part of another group, refer to alkyl, alkenyl, and alkynyl groups as described above with an aryl substituent. Representative examples of arylalkyl include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, phenrthyl, diphenylmethyl, and naphthylmethyl, among others. "Substituted arylalkyl" includes arylalkyl groups in which the aryl portion is optionally substituted with one or more substituents, such as those included in the above definitions of "substituted alkyl" and "substituted cycloalkyl."

[0233] The terms "arylalkyl," "arylalkenyl," and "arylalkynyl," used alone or as part of another group, refer to alkyl, alkenyl, and alkynyl groups as described above with an aryl substituent. Representative examples of arylalkyl include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, phenylethyl, benzhydryl, and naphthylmethyl, among others. "Substituted arylalkyl" includes arylalkyl groups in which the aryl portion is optionally substituted with one or more substituents, such as those included in the above definitions of "substituted alkyl" and "substituted cycloalkyl."

[0234] The term "halogen" or "halo," as used herein alone or as part of another group, refers to chlorine, bromine, fluorine, and iodine.

[0235] The terms "haloalkyl," "haloalkenyl," and "alkynyl," used herein alone or as part of another group, refer to "alkyl," "alkenyl," and "alkynyl" substituted with one or more atoms selected from fluorine, chlorine, bromine, fluorine, and iodine.

[0236] Unless otherwise indicated, the term "aryl" or "Ar", as used herein alone or as part of another group, refers to a monocyclic or polycyclic aromatic group containing 6-10 carbons in the ring portion (e.g., phenyl or naphthyl including 1-naphthyl and 2-naphthyl), and may optionally include one to three additional rings fused to a carbocyclic or heterocyclic ring (e.g., an aryl, cycloalkyl, heteroaryl, or cycloheteroalkyl ring).

[0237] "Substituted aryl" encompasses aryl groups substituted with one or more functional groups such as halogen, haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, alkoxycarbonyl, arylcarbonyl, arylalkenyl, aminocarbonylaryl, arylthio, arylsulfinyl, arylazo, heteroarylalkyl, heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro, Cyano, amino, substituted amino, wherein the amino group contains 1 or 2 substituents which are alkyl, aryl or any other aryl compound mentioned in the definitions, thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino or arylsulfonylaminocarbonyl and / or any alkyl substituent mentioned herein.

[0238] Unless otherwise indicated, the term "heterocyclyl" or "heterocycle" as used herein represents a substituted or unsubstituted stable 5- to 10-membered monocyclic ring system which may be saturated or unsaturated and which consists of carbon atoms and 1-4 heteroatoms selected from N, O or S, wherein the nitrogen and sulfur heteroatoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heterocycle may be attached at any heteroatom or carbon atom that results in a stable structure. Examples of such heterocyclic groups include, but are not limited to, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, oxopyrrolidinyl, oxazepine Oxoazepinyl, aza

[0014] The term "aromatic heterocyclyl" as used herein alone or as part of another group refers to a 5- or 7-membered aromatic ring containing 1, 2, 3 or 4 heteroatoms such as nitrogen, oxygen or sulfur, and wherein such ring is fused to an aryl, cycloalkyl, heteroaryl or heterocycloalkyl ring (e.g., benzothienyl, indolyl), and includes possible N-oxides. "Substituted heteroaryl" includes heteroaryl groups optionally substituted with 1 to 4 substituents, such as those included in the above definitions of "substituted alkyl" and "substituted cycloalkyl". Examples of heteroaryl groups include the following:

[0239] etc.

[0240] Example 1

[0241] 4-Isothiocyanato-2-trifluoromethylbenzonitrile, (1a)

[0242] 4-Amino-2-trifluoromethylbenzonitrile (2.23 g, 12 mmol) was added dropwise to a well-stirred heterogeneous mixture of thiophosgene (1 ml, 13 mmol) in water (22 ml) at room temperature over 15 minutes. The mixture was stirred for an additional hour. The reaction mixture was extracted with chloroform (3 x 15 ml). The combined organic phases were dried over magnesium sulfate and evaporated to dryness under reduced pressure to give the desired product, 4-isothiocyanato-2-trifluoromethylbenzonitrile, (1a), as a brown solid which was used directly in the next step (2.72 g, 11.9 mmol, 99%).

[0243] Example 2

[0244] 2-1). tert-Butyl (4-aminophenyl)carbamate, (2a)

[0245] An aqueous solution of potassium carbonate (1.52 g, 11 mmol in 5 ml of water) was added to a solution of 1,4-phenylenediamine (3.24 g, 30 mmol) in THF (30 ml) and DMF (10 ml). To this mixture was added di-tert-butyl dicarbonate (Boc2O) (2.18 g, 10 mmol) dropwise over 0.5 hours. The reaction mixture was stirred at room temperature for 4 hours. The mixture was then poured into cold water (40 ml) and extracted with chloroform (3 x 50 ml). The combined organic phases were dried over magnesium sulfate and concentrated to give a brown residue, which was purified by flash chromatography (dichloromethane / acetone, 4:1) to give tert-butyl (4-aminophenyl)carbamate, (2a), as a yellow solid (1.98 g, 9.5 mmol, 95%) (yield based on Boc2O).

[0246] 2-2). tert-Butyl {4-[(1-cyano-1-methylethyl)amino]phenyl}carbamate, 2b

[0247] A mixture of 2a (0.83 g, 4 mmol), acetone cyanohydrin (2-methyl-2-hydroxypropionitrile) (4 ml), and MgSO4 (2 g) was heated to 80°C and stirred for 2.5 hours. After cooling to room temperature, compound 2b was crystallized into water (30 ml). The solid was filtered and dried to give tert-butyl {4-[(1-cyano-1-methylethyl)amino]phenyl}carbamate, 2b (1.08 g, 3.9 mmol, 98%).

[0248] 2-3). Tert-butyl {4-[3-(4-cyano-3-trifluoromethylphenyl)-4-imino-5,5-dimethyl-2-thioxo-imidazolidin-1-yl]phenyl}carbamate, (2c)

[0249] Triethylamine (0.202 g, 2 mmol) was added to a solution of 1a (0.456 g, 2 mmol) and 2b (0.57 g, 2 mmol) in dry THF (5 ml). The reaction mixture was stirred at room temperature for 15 hours and then concentrated to give a black residue, which was purified by flash chromatography (ether / acetone, 97:3) to give tert-butyl {4-[3-(4-cyano-3-trifluoromethylphenyl)-4-imino-5,5-dimethyl-2-thioxo-imidazolidin-1-yl]phenyl}carbamate, (2c) (0.15 g, 0.3 mmol, 15%).

[0250] 2-4).4-[3-(4-Aminophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 2d, [RD9]

[0251] A solution of 2c (0.15 g, 0.3 mmol) in 3N aqueous HCl (1 ml) and methanol (4 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (5 ml) and extracted with dichloromethane (8 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane / acetone, 9:1) to afford 4-[3-(4-aminophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 2d, [RD9] (0.118 g, 0.29 mmol, 97%) as a yellow solid.

[0252]

[0253] 1 H NMR(400MHz, CDCl3)δ1.54(s,6H),6.73-6.75(m,2H),7.00-7.03(m,2H),8.0 2(dd,J1=8.2Hz,J2=1.8Hz,1H),8.16(d,J=1.8Hz,1H),8.20(d,J=8.2Hz,1H); 13 C NMR (100MHz, CDCl3) δ22.7,66.2,109.1,114.3,114.9,120.4,122.0(q,J=272.5Hz),127 .0(q,J=4.9Hz),130.4,132.5(q,J=33.0Hz),133.4,135.6,138.5,149.2,175.3,180.4.

[0254] 2-5).4-[3-(4-Azidophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 2e, [RD10]

[0255] Aqueous sulfuric acid (25% wt, 1 mL) was added to a solution of 2d (0.10 g, 0.25 mmol) in acetone (1 mL) at -5°C. An aqueous solution of NaNO₂ (0.024 g, 0.35 mmol in 0.5 mL of water) was slowly added to the mixture over 0.1 hour. The reaction mixture was stirred at -5°C for an additional hour, followed by the dropwise addition of aqueous NaN₃ (0.02 g, 0.3 mmol in 0.3 mL of water). After the addition was complete, the reaction mixture was warmed to room temperature and stirred for an additional 3 hours. The product was extracted with dichloromethane (3 x 5 mL). The combined organic layers were dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-[3-(4-azidophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 2e, [RD10] (0.08 g, 0.18 mmol, 72%) as a light yellow solid.

[0256]

[0257] 1 H NMR(400MHz, CDCl3)δ1.54(s,6H),7.17-7.20(m,2H),7.27-7.30(m,2H),7.8 4(dd,J1=8.3Hz,J2=1.8Hz,1H),7.96(d,J=1.8Hz,1H),7.97(d,J=8.3Hz,1H); 13 C NMR (100MHz, CDCl3) δ23.7,66.4,110.1,114.8,120.4,122.1(q,J=272.5Hz),127.0(q,J =4.7Hz),131.1,131.5,132.3,133.3(q,J=33.0Hz),135.3,137.1,141.7,174.8,180.1.C 19 H 13 MS of F3N6OS, theoretical value 430.4, measured value 430.1.

[0258] Example 3

[0259] 3-1).2-(4-Hydroxyphenylamino)-2-methylpropionitrile, 3a

[0260] 4-Aminophenol (1.09 g, 10 mmol), acetone cyanohydrin (10 ml), and MgSO4 (2 g) were heated to 80°C and stirred for 4 hours. After the mixture was concentrated under vacuum, compound 3a was crystallized from water (20 ml). The solid was filtered and dried to give 2-(4-hydroxyphenylamino)-2-methylpropionitrile, 3a (1.69 g, 9.6 mmol, 96%).

[0261] 3-2).4-[3-(4-Hydroxyphenyl)-5-imino-4,4-dimethyl-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 3b

[0262] Triethylamine (0.101 g, 1 mmol) was added to a solution of 1a (0.456 g, 2 mmol) and 3a (0.352 g, 2 mmol) in dry THF (5 ml). The reaction mixture was stirred at 0°C for 48 hours and then concentrated to give a black residue, which was purified by flash chromatography (dichloromethane / acetone, 85:15) to give 4-[3-(4-hydroxyphenyl)-5-imino-4,4-dimethyl-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 3b (0.274 g, 0.68 mmol, 34%).

[0263] 3-3).4-[3-(4-Hydroxyphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 3c, [RD8]

[0264] A mixture of 3b (0.202 g, 0.5 mmol) in 2N aqueous HCl (2 ml) and methanol (5 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane / acetone, 9:1) to afford 4-[3-(4-hydroxyphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 3c, [RD8] (0.198 g, 0.49 mmol, 98%) as a white powder.

[0265]

[0266] 1 H NMR (CDCl3, 400MHz) δ1.57 (s, 6H), 6.26 (s, OH), 6.90-6.93 (m, 2H), 7.11-7.14 (m, 2H), 7.84 (dd, J1 = 8.3Hz, J2 = 1.8Hz, 1H), 7.95-7.98 (m, 2H); 13 CNMR(CDCl3,100MHz)δ23.6,66.5,109.9,114.9,115.7,116.8,121.9(q,J=272.7Hz),12 7.2(q,J=4.7Hz),130.6,132.3,133.5(q,J=33.2Hz),135.3,137.2,157.0,175.3,180.2.

[0267] Example 4

[0268] 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl]phenyl chloroacetate, 4a, [RD13]

[0269] Chloroacetyl chloride (0.045 g, 0.4 mmol) was added to a solution of 3c (0.101 g, 0.25 mmol) and triethylamine (0.041 g, 0.41 mmol) in dry THF (1.5 ml). The mixture was stirred at room temperature for 4 hours. The triethylamine hydrochloride was filtered off. The filtrate was concentrated and purified by chromatography (dichloromethane / acetone, 95:5) to afford 84% of 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl]phenyl chloroacetate, 4a, [RD13] (0.101 g, 0.21 mmol) as a white powder.

[0270]

[0271] 1 H NMR (CDCl3, 400MHz) δ1.58 (s, 6H), 4.32 (s, 2H), 7.33 (s, 4H), 7.83 (dd, J1 = 8.3Hz, J2 = 1.9Hz, 1H), 7.95-7.97 (m, 2H); 13 C NMR(CDCl3,100MHz)δ23.7,40.8,66.5,110.1,114.8,121.9(q,J=272.5Hz),122.7,127.1(q,J = 4.7Hz), 130.9, 132.3, 132.9, 133.5 (q, J = 33.2Hz), 135.3, 137.1, 150.9, 165.5, 174.8, 180.0.

[0272] Example 5

[0273] 5-1a).2-Methyl-2-(4-methylphenyl)aminopropionitrile, 5a

[0274] A mixture of p-toluidine (1.07 g, 10 mmol) and acetone cyanohydrin (10 ml) was heated to 80°C and stirred for 4 hours. The mixture was concentrated and dried under vacuum to give 2-methyl-2-(4-methylphenyl)aminopropionitrile, 5a (1.72 g, 9.9 mmol, 99%) as a brown solid.

[0275] 5-1b).2-Methyl-2-(4-methylphenyl)aminopropionitrile, 5a

[0276] Sodium cyanide (0.735 g, 15 mmol) was added to a mixture of p-toluidine (1.07 g, 10 mmol) and acetone (1.16 g, 20 mmol) in 90% acetic acid (10 ml). The reaction mixture was stirred at room temperature for 12 hours, after which ethyl acetate (50 ml) was added. The organic layer was washed with water (4 x 30 ml), dried over magnesium sulfate, and concentrated to dryness under vacuum to afford 2-methyl-2-(4-methylphenyl)aminopropionitrile, 5a (1.65 g, 9.5 mmol, 95%) as a brown solid.

[0277] 5-2).4-[3-(4-Methylphenyl)-5-imino-4,4-dimethyl-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 5b

[0278] Triethylamine (0.101 g, 1 mmol) was added to a solution of 1a (0.456 g, 2 mmol) and 5a (0.348 g, 2 mmol) in dry THF (3 ml). The reaction mixture was stirred at 0°C for 2 days and then concentrated to give a black residue, which was purified by flash chromatography (dichloromethane / acetone, 95:5) to give 4-[3-(4-methylphenyl)-5-imino-4,4-dimethyl-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 5b (0.136 g, 0.34 mmol, 17%).

[0279] 5-3a).4-[3-(4-Methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 5c

[0280] A mixture of 5b (0.121 g, 0.3 mmol) in 2N aqueous HCl (2 ml) and methanol (5 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-[3-(4-methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 5c (0.118 g, 0.294 mmol, 98%) as a white powder.

[0281] 5-3b). 4-[3-(4-Methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethyl-benzonitrile, 5c, [RD7]

[0282] A mixture of 1a (0.547 g, 2.4 mmol) and 5a (0.348 g, 2 mmol) in dry DMF (0.6 ml) was stirred for 36 hours. To this mixture was added methanol (20 ml) and 2N HCl (5 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (30 ml) and extracted with ethyl acetate (40 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-[3-(4-methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethyl-benzonitrile, 5c, [RD7] (0.596 g, 1.48 mmol, 74%) as a white powder.

[0283]

[0284] 1 H NMR (CDCl3, 400MHz) δ1.61 (s, 6H), 2.44 (s, 3H), 7.17-7.20 (m, 2H), 7.33-7.36 (m, 2H), 7.86 (dd, J1 = 8.3Hz, J2 = 1.8Hz, 1H), 7.96-7.98 (m, 2H); 13 CNMR (CDCl3, 100MHz) δ21.3, 23.6, 66.4, 110.0, 114.9, 121.9 (q, J = 272.6Hz), 127.1 (q, J = 4. 7Hz), 129.2, 130.6, 132.2, 132.3, 133.4 (q, J = 33.2Hz), 135.2, 137.2, 140.1, 175.1, 179.9.

[0285] Example 6

[0286] 6-1).2-Methyl-2-phenylaminopropionitrile, 6a

[0287] A mixture of aniline (0.931 g, 10 mmol) and acetone cyanohydrin (2 ml) was heated to reflux and stirred for 20 hours. After cooling to room temperature, the reaction mixture was poured into ethyl acetate (40 ml) and washed with cold water (2 x 30 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness under vacuum to provide 2-methyl-2-phenylaminopropionitrile, 6a (1.51 g, 9.4 mmol, 94%) as a brown syrup.

[0288] 6-2).4-[3-phenyl-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 6b, [RD10]

[0289] A mixture of 1a (0.274 g, 1.2 mmol) and 6a (0.160 g, 1 mmol) in dry DMF (0.2 ml) was stirred for 48 hours. Methanol (10 ml) and 2N HCl (3 ml) were added to this mixture. This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-[3-phenyl-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 6b, [RD10] (0.276 g, 0.71 mmol, 71%) as a white powder.

[0290]

[0291] 1 H NMR (CDCl3, 400MHz) δ1.60 (s, 6H), 7.28-7.31 (m, 2H), 7.50-7.58 (m, 3H), 7.85 (dd, J1 = 8.3Hz, J2 = 1.8Hz, 1H), 7.96-7.99 (m, 2H); 13 C NMR(CDCl3,100MHz)δ23.7,66.4,110.2,114.8,121.9(q,J=272.6Hz),127.1(q,J=4.7Hz ), 129.5, 129.8, 129.9, 132.2, 133.4 (q, J = 33.2Hz), 135.1, 135.2, 137.2, 175.0, 179.9.

[0292] Example 7

[0293] 7-1a).1-(4-Methylphenyl)aminocyclobutanenitrile (cyclobutanenitrile), 7a

[0294] Sodium cyanide (0.147 g, 3 mmol) was added to a mixture of p-toluidine (0.214 g, 2 mmol) and cyclobutanone (0.21 g, 3 mmol) in 90% acetic acid (3 mL). The reaction mixture was stirred at room temperature for 12 hours, after which 20 mL of ethyl acetate was added. The organic layer was washed with water (3 x 10 mL), dried over magnesium sulfate, and concentrated to dryness under vacuum to afford 1-(4-methylphenyl)aminocyclobutanecarbonitrile, 7a (0.343 g, 1.84 mmol, 92%) as a brown solid.

[0295] 7-1b).1-(4-Methylphenyl)aminocyclobutanecarbonitrile, 7a

[0296] Trimethylsilyl cyanide (0.93 ml, 7 mmol) was added dropwise to a mixture of p-toluidine (0.535 g, 5 mmol) and cyclobutanone (0.42 g, 6 mmol). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 1-(4-methylphenyl)aminocyclobutanecarbonitrile, 7a (0.912 g, 4.9 mmol, 98%) as a light yellow solid.

[0297] 7-2).4-(8-Imino-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 7b

[0298] To a solution of 1a (2.28 g, 10 mmol) in dry DMF (3 ml) was gradually added a solution of 7a (1.764 g, 9 mmol) in dry DMF (3 ml) at room temperature over 20 hours. The mixture (medium) was stirred for an additional 4 hours. After evaporation of the DMF, the residue was purified by chromatography (dichloromethane / acetone, 95:5) to afford 4-(8-imino-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 7b (1.937 g, 4.68 mmol, 52%).

[0299] 7-3a).4-(8-Oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 7c[RD37]

[0300] A mixture of 7b (0.041 g, 0.1 mmol) in 2N aqueous HCl (3 ml) and methanol (1 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (5 ml) and extracted with ethyl acetate (6 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, and 7c (0.04 g, 0.096 mmol, 96%) as a white powder.

[0301] 7-3b).4-(8-Oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 7c, [RD37]

[0302] A mixture of 1a (0.912 g, 4 mmol) and 7a (0.558 g, 3 mmol) in dry DMF (0.5 ml) was stirred at room temperature for 24 hours. To this mixture was added methanol (30 ml) and 2N aqueous HCl (6 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (50 ml) and extracted with ethyl acetate (60 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 7c (0.959 g, 2.31 mmol, 77%) as a white powder.

[0303]

[0304] 1 H NMR(CDCl3,400MHz)δ1.62-1.69(m,1H),2.16-2.22(m,1H),2.46(s,3H),2.55-2.66(m,4H),7.19-7.26(m ,2H),7.36-7.42(m,2H),7.86(dd,J1=8.3Hz,J2=1.8Hz,1H),7.96(d,J=8.3Hz,1H),7.99(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,21.3,31.4,67.4,109.9,114.9,121.9(q,J=272.6Hz),127.1(q,J= 4.7Hz), 129.5, 130.8, 132.2, 132.4, 133.3 (q, J = 33.2Hz), 135.2, 137.3, 140.1, 175.0, 180.0.

[0305] Example 8

[0306] 8-1).1-(4-Methylphenyl)aminocyclopentanecarbonitrile, 8a

[0307] Trimethylsilyl cyanide (0.865 ml, 7 mmol) was added dropwise to a mixture of p-toluidine (0.535 g, 5 mmol) and cyclopentanone (0.589 g, 7 mmol). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 1-(4-methylphenyl)aminocyclopentanecarbonitrile, 8a (0.981 g, 4.9 mmol, 98%) as a light yellow solid.

[0308] 8-2).4-(4-Oxo-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 8b, [RD35]

[0309] A mixture of 1a (0.296 g, 1.3 mmol) and 8a (0.2 g, 1 mmol) in dry DMF (0.2 ml) was stirred for 48 hours. To this mixture was added methanol (10 ml) and 2N aqueous HCl (3 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-(4-oxo-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 8b, [RD35] (0.3 g, 0.7 mmol, 70%) as a white powder.

[0310]

[0311] 1 H NMR(CDCl3,400MHz)δ1.47-1.57(m,2H),1.81-1.92(m,2H),2.20-2.24(m,2H),2.27-2.34(m,2H),2.43(s,3H),7.1 8-7.22(m,2H),7.33-7.36(m,2H),7.86(dd,J1=8.2Hz,J2=1.8Hz,1H),7.96(d,J=8.2Hz,1H),7.98(d,J=1.8Hz,1H); 13 C NMR (CDCl3, 100MHz) δ21.3, 25.2, 36.3, 75.1, 110.0, 114.9, 121.9 (q, J=272.5Hz), 127.1 (q, J= 4.7Hz), 129.5, 130.7, 123.2, 133.0, 133.4 (q, J = 33.2Hz), 135.1, 137.4, 140.0, 176.3, 180.2.

[0312] Example 9

[0313] 9-1).1-(4-Methylphenyl)aminocyclohexanecarbonitrile, 9a

[0314] Sodium cyanide (0.147 g, 3 mmol) was added to a mixture of p-toluidine (0.214 g, 2 mmol) and cyclohexanone (0.294 g, 3 mmol) in 90% acetic acid (3 mL). The reaction mixture was stirred at room temperature for 12 hours, after which 20 mL of ethyl acetate was added. The organic layer was washed with water (3 x 10 mL), dried over magnesium sulfate, and concentrated to dryness under vacuum to afford 1-(4-methylphenyl)aminocyclohexanecarbonitrile, 9a (0.398 g, 1.86 mmol, 93%) as a brown solid.

[0315] 9-2).4-(4-Imino-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 9b

[0316] Triethylamine (0.05 g, 0.5 mmol) was added to a mixture of 1a (0.228 g, 1 mmol) and 9a (0.214 g, 1 mmol) in dry THF (2 ml). The reaction mixture was stirred at room temperature for 2 days and then concentrated to give a black residue, which was purified by flash chromatography (dichloromethane / acetone, 95:5) to give 4-(4-imino-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 9b (0.035 g, 0.08 mmol, 8%).

[0317] 9-3).4-(4-Oxo-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 9c, [RD48]

[0318] A mixture of 9b (0.035 g, 0.08 mmol) in 2N aqueous HCl (1 ml) and methanol (3 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (5 ml) and extracted with ethyl acetate (6 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 4-(4-oxo-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 9c, [RD48] (0.034 g, 0.076 mmol, 95%) as a white powder.

[0319]

[0320] 1H NMR(CDCl3,400MHz)δ1.02-1.05(m,1H),1.64-1.76(m,4H),2.03-2.12(m,5H),2.44(s,3H),7.12-7.15(m ,2H),7.33-7.36(m,2H),7.85(dd,J1=8.2Hz,J2=1.8Hz,1H),7.96(d,J=8.3Hz,1H),7.97(d,J=1.8Hz,1H); 13 CNMR(CDCl3,100MHz)δ20.7,21.3,24.0,32.6,67.4,109.9,114.9,122.0(q,J=272.5Hz),127.3(q ,J=4.6Hz),130.0,130.5,132.0,132.5,133.3(q,J=33.2Hz),135.2,137.3,140.1,174.1,180.1.

[0321] Example 10

[0322] 10-1).1-(4-Methylphenyl)aminocyclohexanecarbonitrile, 10a

[0323] Sodium cyanide (0.147 g, 3 mmol) was added to a mixture of p-toluidine (0.214 g, 2 mmol) and cyclohexanone (0.337 g, 3 mmol) in 90% acetic acid (3 ml). The reaction mixture was stirred at room temperature for 12 hours, after which 20 ml of ethyl acetate was added. The organic layer was washed with water (3 x 10 ml), dried over magnesium sulfate, and concentrated to dryness under vacuum to afford 1-(4-methylphenyl)aminocyclohexanecarbonitrile, 10a (0.438 g, 1.92 mmol, 96%) as a brown solid.

[0324] 10-2).4-(4-Imino-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]undec-3-yl)-2-trifluoromethylbenzonitrile, 10b

[0325] Triethylamine (0.05 g, 0.5 mmol) was added to a mixture of 1a (0.228 g, 1 mmol) and 9a (0.228 g, 1 mmol) in dry THF (2 ml). The reaction mixture was stirred at room temperature for 2 days and then concentrated to give a black residue, which was purified by flash chromatography (dichloromethane / acetone, 95:5) to give 4-(4-imino-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]undec-3-yl)-2-trifluoromethylbenzonitrile, 10b (0.036 g, 0.08 mmol, 8%).

[0326] 10-3).4-(4-Oxo-2-thioxo-1-(4-methylphenyl)-1,3-diazaspiro[4.5]undec-3-yl)-2-trifluoromethylbenzonitrile, 10c, [RD49]

[0327] A mixture of 9b (0.036 g, 0.08 mmol) in 2N aqueous HCl (1 ml) and methanol (3 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (5 ml) and extracted with ethyl acetate (6 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 10c (0.034 g, 0.075 mmol, 94%) as a white powder.

[0328]

[0329] 1 H NMR(CDCl3,400MHz)δ1.24-134(m,2H),1.37-1.43(m,2H),1.53-1.60(m,2H),1.74-1.82(m,2H),2.19-2.25(m, 4H),2.44(s,3H),7.16-7.19(m,2H),7.32-7.35(m,2H),7.83(dd,J1=8.2Hz,J2=1.8Hz,1H),7.95-7.97(m,2H); 13 C NMR(CDCl3,100MHz)δ21.4,22.2,30.9,36.3,71.1,110.0,114.9,121.9(q,J=272.5Hz),127.2(q ,J=4.6Hz),129.6,130.5,132.3,133.0,133.2(q,J=33.2Hz),135.1,137.4,140.0,175.9,179.7.

[0330] Example 11

[0331] 11-1).1-(4-Hydroxyphenyl)aminocyclobutanecarbonitrile, 11a

[0332] Trimethylsilyl cyanide (0.93 ml, 7 mmol) was added dropwise to a mixture of 4-hydroxyaniline (0.545 g, 5 mmol) and cyclobutanone (0.42 g, 6 mmol). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane:acetone, 98:2) to give 11a (0.903 g, 4.8 mmol, 96%) as a light yellow solid.

[0333] 11-2).4-(8-Oxo-6-thioxo-5-(4-hydroxyphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 11b, [RD58]

[0334] A mixture of 1a (0.57 g, 2.5 mmol) and 7a (0.376 g, 2 mmol) in dry DMF (0.5 ml) was stirred at room temperature for 40 hours. To this mixture was added methanol (30 ml) and aqueous HCl (5 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (40 ml) and extracted with ethyl acetate (50 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 98:2) to afford 11b (0.659 g, 1.58 mmol, 79%) as a white powder.

[0335]

[0336] 1 H NMR(CDCl3,400MHz)δ1.55-1.63(m,1H),2.01-2.09(m,1H),2.50-2.65(m,4H),6.97-7.01(m,2H),7 .20-7.24(m,2H),8.02(dd,J1=8.3Hz,J2=1.8Hz,1H),8.14(d,J=1.8Hz,1H),8.21(d,J=8.3Hz,1H); 13 C NMR (acetone-d6, 100 MHz) δ 13.4, 31.3, 67.5, 108.9, 114.8, 116.1, 123.5 (q, J = 271.5 Hz), 127.4 (q, J = 4.9 Hz), 131.3, 131.8 (q, J = 32.7 Hz), 133.3, 135.5, 136.2, 138.5, 158.1, 175.1, 180.7.

[0337] Example 12

[0338] 12-1).1-(4-Biphenylamino)cyclobutanecarbonitrile, 12a

[0339] Trimethylsilyl cyanide (0.2 ml, 1.5 mmol) was added dropwise to a mixture of 4-biphenylamine (0.169 g, 1 mmol) and cyclobutanone (0.098 g, 1.4 mmol). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 12a (0.24 g, 0.97 mmol, 97%) as a white solid.

[0340] 12-2).4-(8-Oxo-6-thioxo-5-(4-biphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 12b[RD57]

[0341] A mixture of 1a (0.137 g, 0.6 mmol) and 12a (0.124 g, 0.5 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 3 days. To this mixture was added methanol (5 ml) and 2N aqueous HCl (1 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (15 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 12b (0.162 g, 0.34 mmol, 68%) as a white powder.

[0342]

[0343] 1 H NMR(CDCl3,400MHz)δ1.67-1.76(m,1H),2.19-2.31(m,1H),2.59-2.74(m,4H),7.40-7.44(m,3H),7.47-7.53(m,2H),7 .64-7.67(m,2H),7.79-7.82(m,2H),7.88(dd,J1=8.3Hz,J2=1.8Hz,1H),7.97(d,J=8.2Hz,1H),8.02(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.5,67.5,110.0,114.9,122.0(q,J=272.6Hz),127.1(q,J=4.7Hz),127.3,1 28.1,128.7,129.0,130.2,132.3,133.5(q,J=33.2Hz),134.2,135.2,137.2,139.6,142.8,174.9,179.9.

[0344] Example 13

[0345] 13-1).1-(2-Naphthylamino)cyclobutanecarbonitrile, 13a

[0346] Trimethylsilyl cyanide (0.27 ml, 2 mmol) was added dropwise to a mixture of 2-aminonaphthalene (0.143 g, 1 mmol) and cyclobutanone (0.098 g, 1.4 mmol). The reaction mixture was stirred at room temperature for 12 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 13a (0.209 g, 0.94 mmol, 94%) as a yellow solid.

[0347] 13-2).4-(8-Oxo-6-thioxo-5-(4-biphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 12b, [RD85]

[0348] A mixture of 1a (0.137 g, 0.6 mmol) and 13a (0.111 g, 0.5 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 3 days. To this mixture was added methanol (5 ml) and aqueous HCl (1 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (15 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 12b (0.146 g, 0.325 mmol, 65%) as a white powder.

[0349]

[0350] 1 H NMR(CDCl3,400MHz)δ1.58-1.68(m,1H),2.17-2.29(m,1H),2.61-2.75(m,4H),7.40(dd,J1=8.6Hz ,J2=2.0Hz,1H),7.58-7.65(m,2H),7.86-8.00(m,5H),8.04(J=1.8Hz,1H),8.06(d,J=8.6Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.6,67.7,110.0,114.9,122.0(q,J=272.6Hz),126.8,127.1(q,J=4.8Hz),127.2,1 27.7, 128.0, 128.3, 129.1, 130.2, 132.2, 132.5, 133.4, 133.5 (q, J = 33.1Hz), 133.6, 135.2, 137.2, 175.0, 180.1.

[0351] Example 14

[0352] 14-1).2-(4-Methyl-2-pyridylamino)-2-methylpropionitrile, 14a

[0353] Trimethylsilyl cyanide (0.27 ml, 2 mmol) was added dropwise to a mixture of 2-amino-4-methylpyridine (0.108 g, 1 mmol) and acetone (0.58 g, 10 mmol). The reaction mixture was stirred at room temperature for 6 days and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane:acetone, 60:40) to give 14a (0.133 g, 0.76 mmol, 76%) as a white solid.

[0354] 14-2).4-[4,4-Dimethyl-3-(4-methylpyridin-2-yl)-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 14b, [RD83]

[0355] A mixture of 1a (0.91 g, 0.4 mmol) and 14a (0.053 g, 0.3 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 6 days. To this mixture was added methanol (5 ml) and aqueous HCl (1 ml). This second mixture was refluxed for 5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (15 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 14b (0.07 g, 0.174 mmol, 58%) as a white powder.

[0356]

[0357] 1 H NMR (CDCl3, 400MHz) δ1.70 (s, 6H), 2.44 (s, 3H), 7.19 (d, J = 4.4Hz, 1H), 7.45 (t, J = 0.6Hz, 1H), 7.82 (dd, J1=8.2Hz, J2=1.8Hz, 1H), 7.95 (d, J=1.8Hz, 1H), 7.97 (d, J=8.2Hz, 1H), 8.47 (d, J=5.0Hz, 1H); 13 C NMR (CDCl3, 100MHz) δ21.1, 24.1, 67.1, 110.2, 114.8, 121.9 (q, J = 272.6Hz), 124.4, 125.1, 127 .3(q,J=4.8Hz),132.4,133.5(q,J=33.2Hz),135.3,137.1,149.2,149.5,150.0,175.2,179.0.

[0358] Example 15

[0359] 15-1).2-(2-Pyridylamino)-2-methylpropionitrile, 15a

[0360] Trimethylsilyl cyanide (0.27 ml, 2 mmol) was added dropwise to a mixture of 2-aminopyridine (0.094 g, 1 mmol) and acetone (0.58 g, 10 mmol). The reaction mixture was stirred at room temperature for 6 days and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane:acetone, 60:40) to give 15a (0.131 g, 0.81 mmol, 81%) as a white solid.

[0361] 15-2).4-[4,4-Dimethyl-3-(4-pyridin-2-yl)-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 15b, [RD82]

[0362] A mixture of 1a (0.91 g, 0.4 mmol) and 15a (0.048 g, 0.3 mmol) in dry DMF (0.3 ml) was stirred at room temperature for 10 days. To this mixture was added methanol (5 ml) and aqueous HCl (1 ml). This second mixture was refluxed for 5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (15 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 15b (0.059 g, 0.153 mmol, 51%) as a white powder.

[0363]

[0364] 1 H NMR (CDCl3, 400MHz) δ1.73 (s, 6H), 7.38 (dd, J1 = 7.3Hz, J2 = 5.4Hz, 1H), 7.71 (d, J = 8.0Hz, 1H), 7.87 (dd, J1 = 8.2Hz, J2 = 1.8Hz ,1H),7.95(td,J1=7.8Hz,J2=1.8Hz,1H),7.95(d,J=1.3Hz,1H),7.98(d,J=8.2Hz,1H),8.62(dd,J1=4.7Hz,J2=1.3Hz,1H); 13 C NMR(CDCl3,100MHz)δ24.2,67.1,110.3,114.8,121.9(q,J=272.6Hz),123.7,123.8,127.3( q, J=4.8Hz), 132.4, 133.6 (q, J=33.2Hz), 135.3, 137.1, 138.2, 149.5, 149.6, 175.1, 179.0.

[0365] Example 16

[0366] 16-1).1-(5-Methyl-2H-pyrazol-3-ylamino)-cyclobutanecarbonitrile, 16a

[0367] Trimethylsilyl cyanide (0.532 ml, 4.0 mmol) was added dropwise to a mixture of 3-amino-5-methylpyrazole (0.194 g, 2.0 mmol) and cyclobutanone (0.154 g, 2.2 mmol). The reaction mixture was stirred at room temperature for 40 hours and then concentrated under vacuum to give a black liquid, which was purified by chromatography (dichloromethane) to give 16a (0.267 g, 1.52 mmol, 76%) as an off-white powder.

[0368] 16-2).4-[5-(5-methyl-2H-pyrazol-3-yl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-7-yl]-2-trifluoromethyl-benzonitrile, 16b, [RD84]

[0369] A mixture of 1a (0.0684 g, 0.3 mmol) and 16a (0.053 g, 0.3 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 4 days. To this mixture was added methanol (10 ml) and 2N aqueous HCl (2 ml). This second mixture was refluxed for 5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (30 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 97:3) to afford 16b (0.0826 g, 0.2 mmol, 67%) as a white powder.

[0370]

[0371] 1 H NMR (acetone d6, 400 MHz) δ 1.66-1.76 (m, 1H), 2.00-2.07 (m, 1H), 3.35 (s, 3H), 2.56-2.63 (m, 2H), 2.85-2.93 (m, 2H), 8.04 (dd, J1 = 8.2 Hz, J2 = 1.6 Hz, 1H), 8.18 (d, J = 1.6 Hz, 1H), 8.22 (d, J = 8.2 Hz, 1H), 11.99 (s, 1H); 13C NMR (acetone d6, 100 MHz) δ 10.2, 13.1, 31.1, 67.4, 102.5, 109.1, 114.8, 122.5 (q, J = 271.4 Hz), 127.8 (q, J = 4.8 Hz), 131.9 (q, J = 33.6 Hz), 133.6, 135.6, 138.4, 139.9, 145.0, 175.0, 179.6.

[0372] Example 17

[0373] 4-[3-(4-Hydroxyphenyl)-4,4-dimethyl-2,5-dithioimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 17a, [RD59]

[0374] A mixture of 3c (0.081 g, 0.2 mmol) and Lawesson's reagent (0.097 g, 0.24 mmol) in toluene (3 mL) was heated to reflux for 15 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 mL) and extracted with ethyl acetate (10 mL). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (9:1 dichloromethane:pentane) to afford 17a (0.0185 g, 0.044 mmol, 22%) as a white powder.

[0375]

[0376] 1 H NMR(CDCl3,400MHz)δ1.65(s,6H),6.95-6.97(m,2H),7.15-7.18(m,2H),7.7 5(d,J=8.2Hz,1H),7.86(d,J=1.8Hz,1H),7.98(dd,J1=8.2Hz,J2=1.8Hz,1H); 13 C NMR (CDCl3, 100MHz) δ27.9,77.8,110.9,114.7,116.7,121.9(q,J=272.6Hz),128.1(q,J = 4.8Hz), 129.1, 130.7, 133.3, 133.5 (q, J = 33.2Hz), 135.5, 140.3, 156.8, 179.9, 207.9.

[0377] Example 18

[0378] 4-[3-(4-Hydroxyphenyl)-4,4-dimethyl-2,5-dioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile, 18a, [RD60]

[0379] 30% hydrogen peroxide (3 mL, 26 mmol) was added dropwise to a solution of 3c (0.121 g, 0.4 mmol) in glacial acetic acid (3 mL). The mixture was stirred at room temperature for 12 hours, after which 20 mL of ethyl acetate was added. The organic layer was washed with water (3 x 15 mL), dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 18a (0.102 g, 0.261 mmol, 87%) as a white powder.

[0380]

[0381] 1 H NMR(CDCl3,400MHz)δ1.52(s,6H),6.70-6.73(m,2H),7.01-7.04(m,2H),7.9 2(d,J=8.4Hz,1H),8.00(dd,J1=8.4Hz,J2=1.8Hz,1H),8.15(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ23.7,63.7,108.4,115.0,116.7,121.9(q,J=272.6Hz),123.5(q,J = 4.8Hz), 124.0, 128.5, 130.5, 133.6 (q, J = 33.2Hz), 135.5, 136.2, 153.4, 157.2, 174.5.

[0382] Example 19

[0383] 19-1).3-Fluoro-2-methyl-2-(4-methylphenyl)aminopropionitrile, 19a

[0384] Trimethylsilyl cyanide (0.146 ml, 1.1 mmol) was added dropwise to a mixture of p-toluidine (0.107 g, 1 mmol) and fluoroacetone (0.082 g, 1.1 mmol). The reaction mixture was stirred at room temperature for 12 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 19a (0.179 g, 0.93 mmol, 93%) as a light yellow solid.

[0385] 19-2).4-(4-Fluoromethyl-4-methyl-5-oxo-2-thioxo-3-(4-methylphenyl)imidazolidin-1-yl)-2-trifluoromethylbenzonitrile, 19b, [RD68]

[0386] A mixture of 1a (0.16 g, 0.7 mmol) and 19a (0.096 g, 0.5 mmol) in dry DMF (0.3 ml) was stirred at room temperature for 48 hours. To this mixture was added methanol (10 ml) and 2N aqueous HCl (2 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (30 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 19b (0.168 g, 0.4 mmol, 80%) as a white powder.

[0387]

[0388] 1 H NMR(CDCl3,400MHz)δ1.49(s,3H),2.44(s,3H),4.35(dd,J1=47.2Hz,J2=10.0Hz,1H),4.71(dd,J1=45.2Hz,J2=10Hz,1H) ,7.22-7.26(m,2H),7.35-7.39(m,2H),7.82(dd,J1=8.2Hz,J2=1.8Hz,1H),7.93(d,J=1.8Hz,1H),7.98(d,J=8.2Hz,1H); 13 CNMR(CDCl3,100MHz)δ17.0(d,J=4.6Hz),21.3,69.3(d,J=18.3Hz),81.9(d,J=179.5Hz),109.9,114.8,121.8(q,J= 272.6Hz), 127.2 (q, J = 4.7Hz), 129.3, 130.9, 131.6, 132.3, 133.3 (q, J = 33.2Hz), 135.3, 137.0, 140.5, 174.1, 181.4; 19 F NMR (CDCl3, 376MHz) δ-62.5, 110.9.

[0389] Example 20

[0390] 20-1).2-Methyl-2-(4-trifluoromethylphenyl)aminopropionitrile, 20a

[0391] A mixture of 4-trifluoromethylaniline (1.61 g, 10 mmol), acetone cyanohydrin (5 ml), and magnesium sulfate (2 g) was heated to 80°C and stirred for 12 hours. Ethyl acetate (50 ml) was added to the mixture, which was then washed with water (3 x 30 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness under vacuum to afford compound 20a (2.166 g, 9.5 mmol, 95%) as a brown solid.

[0392] 20-2).4-(4,4-Dimethyl-5-oxo-2-thioxo-3-(4-trifluoromethylphenyl)imidazolidin-1-yl)-2-trifluoromethylbenzonitrile, 20b, [RD66]

[0393] A mixture of 1a (0.114 g, 0.5 mmol) and 20a (0.092 g, 0.4 mmol) in dry DMF (0.3 ml) was stirred at room temperature for 48 hours. To this mixture was added methanol (10 ml) and aqueous HCl (3 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (20 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 20b (0.117 g, 0.256 mmol, 64%) as a white powder.

[0394]

[0395] 1 H NMR(CDCl3,400MHz)δ1.61(s,6H),7.45-7.49(m,2H),7.80-7.83(m,2H),7.8 5(dd,J1=8.3Hz,J2=1.8Hz,1H),7.97(d,J=1.8Hz,1H),7.99(d,J=8.2Hz,1H); 13 C NMR(CDCl3,100MHz)δ23.8,66.6,110.3,114.8,121.8(q,J=272.6Hz),123.5(q,J=271.1Hz),127.0(q,J=4.6Hz), 127.1 (q, J = 4.7Hz), 130.3, 131.9 (q, J = 32.9Hz), 132.2, 133.5 (q, J = 33.3Hz), 135.3, 136.9, 138.4, 174.6, 179.9.

[0396] Example 21

[0397] 21-1).3-Chloro-2-chloromethyl-2-(4-methylphenyl)aminopropionitrile, 21a

[0398] Trimethylsilyl cyanide (0.27 ml, 2 mmol) was added dropwise to a mixture of p-toluidine (0.107 g, 1 mmol) and 1,3-dichloroacetone (0.254 g, 2 mmol). The reaction mixture was heated to 80°C and stirred for 6 hours. To this mixture was added 20 ml of ethyl acetate, which was then washed with water (2 x 20 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 21a (0.192 g, 0.79 mmol, 79%) as a brown powder.

[0399] 21-2).4-(4,4-Bischloromethyl-5-oxo-2-thioxo-3-(4-methylphenyl)imidazolidin-1-yl)-2-trifluoromethylbenzonitrile, 21b, [RD67]

[0400] A mixture of 1a (0.16 g, 0.7 mmol) and 21a (0.122 g, 0.5 mmol) in dry DMF (0.5 ml) was stirred at room temperature for 10 days. To this mixture was added methanol (10 ml) and 2N aqueous HCl (2 ml). The second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 21b (0.09 g, 0.19 mmol, 38%) as a white powder.

[0401]

[0402] 1 H NMR(CDCl3,400MHz)δ2.44(s,3H),3.54(d,J=11.8Hz,2H),3.93(d,J=11.8Hz,2H),7.37-7.40(m,2H) ,7.48-7.51(m,2H),7.79(dd,J1=8.2Hz,J2=1.8Hz,1H),7.88(d,J=1.8Hz,1H),7.98(d,J=8.2Hz,1H); 13 C NMR (CDCl3, 100MHz) δ21.4, 42.8, 74.3, 110.7, 114.7, 121.7 (q, J = 272.6Hz), 127.2 (q, J = 4. 7Hz), 128.8, 131.0, 131.1, 132.4, 133.8 (q, J = 33.2Hz), 135.5, 136.9, 140.9, 169.5, 182.5.

[0403] Example 22

[0404] 22-1).1-(4-Methylphenyl)aminocyclohexanecarbonitrile, 22a

[0405] Sodium cyanide (0.245 g, 5 mmol) was added to a mixture of anthranilic acid (0.411 g, 3 mmol) and acetone (1 ml, 13.6 mmol) in 90% acetic acid (3 ml). The reaction mixture was stirred at room temperature for 12 hours, after which 50 ml of ethyl acetate was added. The organic layer was washed with brine (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 90:10) to afford compound 22a (0.551 g, 2.7 mmol, 90%) as a brown solid.

[0406] 22-2).2-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl]benzoic acid, 22b, [RD65]

[0407] A mixture of 1a (0.114 g, 0.1 mmol) and 22a (0.103 g, 0.5 mmol) in dry DMF (0.5 ml) was stirred at room temperature for 3 days. To this mixture was added methanol (10 ml) and 2N aqueous HCl (3 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (ethyl acetate:pentane, 2:1) to afford 22b (0.143 g, 0.33 mmol, 66%) as a white powder.

[0408]

[0409] 1 H NMR (CDCl3, 400MHz) δ1.47 (s, 3H), 1.78 (s, 3H), 7.39 (d, J = 7.7Hz, 1H), 7.63 (t, J = 7. 7Hz,1H)7.76-7.82(m,2H),7.90-7.98(m,2H),8.22(d,J=6.8Hz,1H),8.96(bs,1H); 13 C NMR (CDCl3, 100MHz) δ20.6, 26.2, 67.6, 110.1, 114.8, 121.9 (q, J = 272.6Hz), 127.2 (q, J = 4.7Hz), 128. 9,131.0,130.2,132.5,133.2(q,J=33.3Hz),133.7,134.7,135.4,135.8,137.3,169.8,175.3,180.7.

[0410] Example 23

[0411] 23-1).1-(2-Methylphenyl)aminocyclobutanecarbonitrile, 23a

[0412] Trimethylsilyl cyanide (0.66 ml, 5 mmol) was added dropwise to a mixture of p-toluidine (0.321 g, 3 mmol) and cyclobutanone (0.28 g, 4 mmol). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 23a (0.541 g, 2.91 mmol, 97%) as a light yellow solid.

[0413] 23-2).4-(8-Oxo-6-thioxo-5-(2-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 23b, [RD71]

[0414] A mixture of 1a (0.114 g, 0.5 mmol) and 23a (0.093 g, 0.5 mmol) in dry DMF (0.3 ml) was stirred at room temperature for 3 days. To this mixture was added methanol (10 ml) and 2N aqueous HCl (3 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 23b (0.116 g, 0.28 mmol, 56%) as a white powder.

[0415]

[0416] 1 H NMR(CDCl3,400MHz)δ1.63-1.69(m,1H),2.26(s,3H),2.28-2.41(m,2H),2.58-2.76(m,3H),7.21(d,J=7.6H z,1H),7.39-7.49(m,3H),7.89(dd,J1=8.2Hz,J2=1.8Hz,1H),7.97(d,J=8.2Hz,1H),8.00(d,J=1.8Hz,1H); 13CNMR(CDCl3,100MHz)δ14.2,18.0,30.7,32.2,67.6,109.9,114.9,121.9(q,J=272.6Hz),127.0(q,J=4.7Hz) ,127.5,129.8,130.2,131.9,132.3,133.4,133.5(q,J=34.3Hz),135.2,135.8,137.1,138.0,175.3,178.7.

[0417] Example 24

[0418] 24-1).1-Aminocyclopentanecarbonitrile, 24a

[0419] Anhydrous ammonia was bubbled into a mixture of cyclopentanone (0.452 g) and trimethylsilyl cyanide (0.66 ml, 5 mmol). Excess ammonia was refluxed through a dry ice-acetone condenser. After refluxing for 1 hour, the ammonia was removed from the mixture, and the remaining mixture was concentrated under vacuum to yield compound 24a (0.522 g, 4.75 mmol, 95%) as a colorless liquid.

[0420] 24-2).4-(4-Imino-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 24b

[0421] Triethylamine (0.101 g, 0.1 mmol) was added to a mixture of 1a (0.684 g, 3 mmol) and 24a (0.33 g, 3 mmol) in dry THF (5 ml). The reaction mixture was stirred at room temperature for 5 hours and then concentrated to give a brown residue, which was purified by flash chromatography (dichloromethane / acetone, 93:7) to give 24b (0.741 g, 2.19 mmol, 73%).

[0422] 24-3).4-(4-Oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 24c, [RD77]

[0423] A mixture of 24b (0.741 g, 2.19 mmol) in 2N aqueous HCl (4 ml) and methanol (20 ml) was heated to reflux for 1 hour. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (40 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 24c (0.72 g, 2.12 mmol, 97%) as a white powder.

[0424]

[0425] 1 H NMR(CDCl3,400MHz)δ1.86-190(m,2H),1.96-2.05(m,4H),2.26-2.30(m,2H),7.80(dd ,J1=8.2Hz, J2=1.8Hz,1H),7.92(d,J=1.8Hz,1H),7.97(d,J=8.2Hz,1H)8.20(bs,NH); 13 C NMR(CDCl3,100MHz)δ25.3,38.1,71.0,110.1,114.8,121.8(q,J=272.7Hz), 126.8 (q, J=4.7Hz), 131.9, 133.6 (q, J=34.3Hz), 135.3, 136.7, 176.1, 179.8.

[0426] Example 25

[0427] 25).4-[1-(4-nitrophenyl)-4-oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl]-2-trifluoromethylbenzonitrile, 25a, [RD55]

[0428] A mixture of 25c (0.0678 g, 0.2 mmol), 1,8-diazabicyclo[5.4.0]undec-7-ene (0.05 g, 0.33 mmol), and 4-fluoronitrobenzene (0.056 g, 0.4 mmol) in dimethylformamide (0.5 ml) was placed in a sealed tube under argon and heated to 130°C for 40 hours. The reaction mixture was poured into ethyl acetate (5 ml) and washed with water (2 x 10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 25a (0.038 g, 0.084 mmol, 42%) as a white powder.

[0429]

[0430] 1 H NMR(CDCl3,400MHz)δ1.53-1.56(m,2H),1.90-1.93(m,2H),2.14-2.18(m,2H),2.37-2.40(m,2H),7.54-7.57 (m,2H),7.85(dd,J1=8.2Hz,J2=1.8Hz,1H),7.97(d,J=1.8Hz,1H),7.98(d,J=8.2Hz,1H),8.39-8.43(m,2H); 13CNMR(CDCl3,100MHz)δ25.2,36.5,75.3,110.3,114.8,121.8(q,J=272.6Hz),125.2,127.0( q,J=4.7Hz),131.4,132.1,133.6(q,J=34.3Hz),135.3,136.9,141.7,148.1,175.6,180.2.

[0431] Example 26

[0432] 26).4-[1-(4-Cyanophenyl)-4-oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl]-2-trifluoromethylbenzonitrile, 26a, [RD54]

[0433] A mixture of 24c (0.0678 g, 0.2 mmol), 1,8-diazabicyclo[5.4.0]dodec-7-ene (0.061 g, 0.4 mmol), and 4-fluorocyanobenzene (0.048 g, 0.4 mmol) in dimethylformamide (0.5 ml) was placed in a sealed tube under argon and heated to 140°C for 5 days. The reaction mixture was poured into ethyl acetate (5 ml) and washed with water (2 x 10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 26a (0.023 g, 0.052 mmol, 26%) as a white powder.

[0434]

[0435] 1 H NMR(CDCl3,400MHz)δ1.51-1.55(m,2H),1.90-1.93(m,2H),2.12-2.16(m,2H) ,2.33-2.38(m,2H),7.47-7.50(m,2H),7.81-7.87(m,3H),7.95-7.99(m,2H); 13 C NMR (CDCl3, 100MHz) δ25.2, 36.5, 75.3, 110.3, 113.9, 114.7, 117.5, 121.8 (q, J = 272.6Hz), 127 .0(q,J=4.8Hz),131.2,132.1,133.6(q,J=34.3Hz),133.8,135.3,136.9,140.0,175.6,180.1.

[0436] Example 27

[0437] 27-1).1-Methyl-4-(4-methylphenylamino)piperidine-4-carbonitrile, 27a

[0438] Sodium cyanide (0.318 g, 6.5 mmol) was added to a mixture of p-toluidine (0.535 g, 5 mmol) and 1-methyl-4-piperidone (0.678 g, 6 mmol) in 90% acetic acid (5 mL). The reaction mixture was stirred at room temperature for 6 hours, after which 100 mL of dichloromethane was added. The organic layer was washed with 2N NaOH solution (2 x 50 mL), dried over magnesium sulfate, concentrated, and purified by chromatography (DCM then acetone) to afford 27a (0.722 g, 3.15 mmol, 63%).

[0439] 27.2).4-(4-Imino-8-methyl-2-thioxo-1-(4-methylphenyl)-1,3,8-triazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 27b

[0440] Triethylamine (0.02, 0.2 mmol) was added to a mixture of 1a (0.228 g, 1 mmol) and 27a (0.114 g, 0.5 mmol) in dry THF (2 ml). The reaction mixture was stirred at room temperature for 20 hours and then concentrated to give a black residue, which was purified by flash chromatography (dichloromethane / acetone, 90:10, then acetone) to give 27b (0.059 g, 0.13 mmol, 26%).

[0441] 27-3).4-(8-Methyl-4-oxo-2-thioxo-1-(4-methylphenyl)-1,3,8-triazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 27c, [RD53]

[0442] A mixture of 27b (0.059 g, 0.13 mmol) in 2N aqueous HCl (1 ml) and methanol (3 ml) was heated to reflux for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (5 ml) and extracted with ethyl acetate (10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 60:40) to afford 27c (0.055 g, 0.012 mmol, 92%) as a white powder.

[0443]

[0444] 1H NMR (acetone-d6, 400 MHz) δ 1.93-1.99 (m, 1H), 2.00-2.04 (m, 1H), 2.18 (s, 3H), 2.24-2.28 (m, 2H), 2.38 (s, 3H), 2.61-2.72 (m, 4H), 7.18-7-20 (m, 2H), 7.32-7.35 (m, 2H), 8.03 (dd, J1=8.2 Hz, J2=1.8 Hz, 1H), 8.16 (d, J=1.8 Hz, 1H), 8.22 (d, J=8.2 Hz, 1H); 13 CNMR (acetone-d6, 100 MHz) δ 20.3, 31.4, 45.1, 49.8, 65.1, 109.1, 114.8, 122.4 (q, J = 275.1 Hz), 127.7 (q, J = 4.8 Hz), 130.0, 130.5, 131.9 (q, J = 32.6 Hz), 132.6, 133.5, 135.6, 138.3, 139.4, 174.0, 180.6.

[0445] Example 28

[0446] 4-(8-Methyl-4-oxo-2-thioxo-1,3,8-triazaspiro[4.5]dec-3-yl)-2-trifluoromethylbenzonitrile, 28a, [RD52]

[0447] Compound 28a was synthesized according to the method described in patent US 5958936.

[0448]

[0449] 1 H NMR (acetone-d6, 400 MHz) δ 1.93-2.00 (m, 2H), 2.09-2.16 (m, 2H), 2.25 (s, 3H), 2.42-2.49 (m, 2H), 2.75-2.80 (m, 2H), 7.97 (dd, J1=8.2 Hz, J2=1.8 Hz, 1H), 8.11 (d, J=1.8 Hz, 1H), 8.20 (d, J=8.2 Hz, 1H), 9.80 (bs, NH); 13 C NMR (acetone-d6, 100 MHz) δ 32.9, 45.4, 50.1, 62.3, 109.1, 114.8, 122.4 (q, J = 271.6 Hz), 127.5 (q, J = 4.8 Hz), 131.8 (q, J = 32.7 Hz), 133.2, 135.6, 135.6, 138.0, 175.2, 180.4.

[0450] Example 29

[0451] 4-[3-(4-Hydroxybutyl)-4,4-dimethyl-5-oxo-2-imidazolidin-1-ylthioxo]-2-trifluoromethylbenzonitrile, RU 59063

[0452] Compound RU 59063 was synthesized according to the method described by Teutsch et al. [J. Steroid. Biochem. Molec. Biol. 1994, 48(1), 111-119].

[0453]

[0454] 1 H NMR(CDCl3,400MHz)δ1.55(s,6H),1.58-1.62(m,2H),1.86-1.89(m,2H),2.25(bs,OH),3.65- 3.71(m,4H),7.74(dd,J1=8.0Hz,J2=1.8Hz,1H),7.92(d,J=1.8Hz,1H),7.98(d,J=8.0Hz,1H); 13 C NMR(CDCl3,100MHz)δ23.1,24.7,29.6,43.9,61.7,65.2,109.7,114.9,121.9(q,J=27 2.6Hz), 127.1 (q, J = 4.8Hz), 132.2, 133.7 (q, J = 34.3Hz), 135.2, 137.2, 175.3, 178.2.

[0455] Example 30

[0456] 30-1).1-Methylaminocyclobutanecarbonitrile, 30a

[0457] Methylamine was bubbled into a cooled mixture of cyclobutanone (0.21 g, 3 mmol) and trimethylsilyl cyanide (0.396 g, 4 mmol) until the volume doubled. The mixture was stirred for 3 h and then concentrated to dryness to afford 30a (0.33 g, quantitative).

[0458] 30-2).4-(5-methyl-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile (enzonitrile), 30b, [RD73]

[0459] A mixture of 1a (0.114 g, 0.5 mmol) and 30a (0.055 g, 0.5 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 0.5 h. To this mixture was added 10 ml of methanol and 2 ml of 2N HCl. This second mixture was refluxed for 2 h. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 30b (0.148 g, 0.435 mmol, 87%) as a white powder.

[0460]

[0461] 1 H NMR(CDCl3,400MHz)δ1.95-2.06(m,1H),2.21-2.32(m,1H),2.58-2.71(m,4H),3.44(s ,3H),7.77(dd,J1=8.2Hz,J2=2.0Hz,1H),7.89(d,J=2.0Hz,1H),7.93(d,J=8.2Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,30.3,30.4,66.1,109.7,114.9,121.9(q,J=272.6Hz ), 126.9 (q, J = 4.8Hz), 132.1, 133.2 (q, J = 34.3Hz), 135.2, 137.3, 175.1, 178.7.

[0462] 30-3).4-(5-Methyl-6,8-dioxo-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile, 30c, [RD74]

[0463] Hydrogen peroxide (2 mL, 30%) was added to a mixture of 30b (0.068 g, 0.2 mmol) in glacial acetic acid (3 mL). After stirring at room temperature for 10 hours, the reaction mixture was poured into ethyl acetate (20 mL) and washed with water (2 x 20 mL). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone) to afford 30c (0.057 g, 0.176 mmol, 88%) as a white powder.

[0464]

[0465] 1H NMR(CDCl3,400MHz)δ1.91-2.35(m,1H),2.21-2.31(m,1H),2.50-2.61(m,4H),3.12(s, 3H),7.89(d,J=8.2Hz,1H),7.97(dd,J1=8.2Hz,J2=2.0Hz,1H),8.12(d,J=2.0Hz,1H),; 13 C NMR(CDCl3,100MHz)δ13.9,25.4,29.3,63.4,108.1,115.1,121.6(q,J=272.6Hz ), 122.9 (q, J = 4.8Hz), 127.9, 133.5 (q, J = 34.3Hz), 135.3, 136.5, 152.7, 174.4.

[0466] Example 31

[0467] 31-1).1-Methylaminocyclopentanecarbonitrile, 31a

[0468] Methylamine was bubbled into a chilled mixture of cyclopentanone (0.252 g, 3 mmol) and trimethylsilyl cyanide (0.396 g, 4 mmol) until the volume doubled. The mixture was stirred for 3 h and then concentrated to dryness to afford 31a (0.372 g, quantitative).

[0469] 31-2).4-(1-methyl-4-oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 31b, [RD75]

[0470] A mixture of 1a (0.114 g, 0.5 mmol) and 31a (0.062 g, 0.5 mmol) in dry DMF (0.2 ml) was stirred at room temperature for 0.5 h. To this mixture was added 10 ml of methanol and 2 ml of 2N HCl. This second mixture was refluxed for 2 h. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 31b (0.159 g, 0.45 mmol, 90%) as a white powder.

[0471]

[0472] 1H NMR(CDCl3,400MHz)δ1.91-2.05(m,6H),2.16-2.21(m,2H),3.27(s,3H),7.7 7(dd,J1=8.2Hz,J2=1.8Hz,1H),7.89(d,J=1.8Hz,1H),7.91(d,J=8.2Hz,1H); 13 C NMR(CDCl3,100MHz)δ26.4,30.3,35.4,73.2,109.5,114.9,121.9(q,J=272.6Hz ), 126.9 (q, J = 4.8Hz), 132.2, 133.2 (q, J = 34.3Hz), 135.2, 137.5, 176.8, 178.5.

[0473] 31-3).4-(1-Methyl-2,4-dioxo-1,3-diaza-spiro[4.4]nonan-3-yl)-2-trifluoromethylbenzonitrile, 31c, [RD76]

[0474] Hydrogen peroxide (2 mL, 30%) was added to a mixture of 31b (0.07 g, 0.2 mmol) in glacial acetic acid (3 mL). After stirring at room temperature for 10 hours, the reaction mixture was poured into ethyl acetate (20 mL) and washed with water (2 x 20 mL). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone) to afford 31c (0.057 g, 0.168 mmol, 84%) as a white powder.

[0475]

[0476] 1 H NMR(CDCl3,400MHz)δ1.88-1.99(m,6H),2.12-2.17(m,2H),2.98(s,3H),7.8 8(d,J=8.2Hz,1H),7.97(dd,J1=8.2Hz,J2=1.8Hz,1H),8.12(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ25.2,26.5,34.8,70.1,108.0,115.1,122.0(q,J=272.5Hz ), 122.9 (q, J = 4.9Hz), 127.9, 133.5 (q, J = 32.9Hz), 135.3, 136.6, 152.7, 176.1.

[0477] Example 32

[0478] 4-(8-Methylimino-6-thioxo-5-p-tolyl-5,7-diaza-spiro[3.4]octan-7-yl)-2-trifluoromethyl-benzonitrile, 32a, [RD90]

[0479] A mixture of 7b (0.042 g, 0.1 mmol), DBU (0.023 g, 0.15 mmol) and iodomethane (0.073 g, 0.5 mmol) in DMF (0.3 ml) was stirred at room temperature for 15 hours. After evaporation of the DMF, the residue was purified by chromatography (dichloromethane) to afford 32a (0.011 g, 0.026 mmol, 26%) as a white powder.

[0480]

[0481] 1 H NMR (CDCl3, 400MHz) δ1.58-1.65 (m, 1H), 2.04-2.13 (m, 1H), 2.45 (s, 3H), 2.70-2.77 (m, 2H), 3.06-3.10 (m, 2H), 3.58 (s, CH3-N, major isomer) [2.70 (s, CH3-N, minor isomer)], 7.20-7.34 (m, 4H), 7.75-7.91 (m, 3H); (C DCl3, 100 MHz) δ 12.6, 21.4, 30.2, 33.7 (35.3 other isomers), 66.9, 109.1, 115.2, 122.1 (q, J = 272.5 Hz), 128.5 (q, J = 4.9 Hz), 129.8, 130.4, 130.6, 132.8, 133.2 (q, J = 32.9 Hz), 133.5, 134.9, 139.8, 157.0, 180.2.

[0482] Example 33

[0483] 1-[3-(4-cyano-3-trifluoromethyl-phenyl)-5,5-dimethyl-2-thioxo-1-p-tolyl-imidazolidin-4-ylidene]-3-ethyl-thiourea, 33a, [RD91]

[0484] A mixture of 5b (0.06 g, 0.149 mmol), ethylthioisocyanate (0.087 g, 1 mmol), and CuI (0.01 g, 0.05 mmol) in DMF (0.1 ml) was heated under microwave for 45 minutes. The mixture was then washed with brine and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (HPLC, alumina column) to afford 33a (0.054 g, 0.108 mmol, 72%) as a white powder.

[0485]

[0486] 1 H NMR (CDCl3, 400 MHz) δ 1.15 (t, J = 7.23 Hz, 3H), 1.70 [1.75 minor isomer] (s, 6H), 2.42 (s, 3H), 3.28-3.39 (m, 2H) [3.15-3.22 (m, 2H), minor isomer], 6.50 (bs, 1H) [6.93 (bs, 1H), minor isomer], 7.14-7.18 (m, 2H), 7.32-7.35 (m, 2H), 7.77-7.94 (m, 3H); 13 C NMR (CDCl3, 100 MHz) δ 13.31 (13.83 minor), 21.3, 25.22 (24.89 minor), 40.31 (40.67 minor), 68.1, 109.9, 114.9, 122.3 (q, J = 272.5 Hz), 127.6 (q, J = 4.9 Hz), 129.1, 129.59 (129. 55 minor), 130.52 (130.57 minor), 132.27 (132.15 minor), 132.9 (q, J = 32.9 Hz), 134.27 (134.15 minor), 134.9, 135.2, 156.33 (156.06 minor), 180.28 (180.06 minor), 187.24 (186.63 minor).

[0487] Example 34

[0488] 1-[7-(4-Cyano-3-trifluoromethyl-phenyl)-6-thioxo-5-p-tolyl-5,7-diaza-spiro[3.4]octan-8-ylidene]-3-phenyl-thiourea, 34a, [RD92]

[0489] A mixture of 7b (0.021 g, 0.05 mmol) and phenylthioisocyanate (0.027 g, 0.2 mmol) in DMF (0.3 ml) was stirred at 60° C. for 2 days. After evaporation of DMF, the mixture was purified by chromatography (dichloromethane) to give 34a (0.015 g, 0.028 mmol, 57%) as a white powder.

[0490]

[0491] 1H NMR(CDCl3,400MHz)δ1.59-1.67(m,1H),2.12-2.22(m,1H),2.45(s,3H),2.61-2.71(m,2H) ,2.81-2.87(m,2H),7.18-7.27(m,6H),7.33-7.41(m,5H),7.60-7.62(m,1H),8.40(bs,1H); 13 C NMR(CDCl3,100MHz)δ13.6,21.4,32.3,69.6,110.7,114.8,121.6,122.0(q,J=272.5Hz),126.3,128.0(q,J=4.9 Hz), 128.9, 129.4, 130.7, 132.5, 133.2 (q, J = 32.9Hz), 134.1, 134.9, 137.7, 139.2, 140.2, 154.8, 180.3, 185.5.

[0492] Example 35

[0493] 1-(4-Cyano-3-trifluoromethyl-phenyl)-3-[7-(4-Cyano-3-trifluoromethyl-phenyl)-6-thioxo-5-p-tolyl-5,7-diaza-spiro[3.4]octan-8-ylidene]-thiourea, 35a, [RD93]

[0494] A mixture of 1a (0.5.02 g, 2.2 mmol) and 7a (0.186 g, 1 mmol) in DMF (1 ml) was stirred at room temperature. After stirring for 20 hours, the mixture was concentrated under reduced pressure to give an orange viscous liquid, which was purified by chromatography (dichloromethane:acetone, 99:1) to give 35a (0.269 g, 0.42 mmol, 42%) as a yellow powder.

[0495]

[0496] X-ray structure of 35a

[0497] Example 36

[0498] 36-1).1-(4-Hydroxymethylphenylamino)-cyclobutanecarbonitrile, 36a

[0499] Trimethylsilyl cyanide (0.66 ml, 5 mmol) was added dropwise to a mixture of 4-aminobenzoic acid (0.492 g, 4 mmol) and cyclobutanone (0.35 g, 5 mmol) in dichloromethane (10 ml). The reaction mixture was stirred at room temperature for 6 hours and then concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane) to give 36a (0.677 g, 3.36 mmol, 84%) as a brown solid.

[0500] 36-2).4-[8-(4-Hydroxymethylphenyl)-5-oxo-7-thioxo-6-azaspiro[3.4]octan-6-yl]-2-trifluoromethyl-benzonitrile, 36b, [RD110]

[0501] A mixture of 1a (0.342 g, 1.5 mmol) and 36a (0.21 g, 1 mmol) in dry DMF (0.5 ml) was stirred at room temperature for 24 hours. To this mixture was added methanol (20 ml) and 2N aqueous HCl (5 ml). This second mixture was refluxed for 6 hours. After cooling to room temperature, the reaction mixture was poured into cold water (40 ml) and extracted with ethyl acetate (60 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 90:10) to afford 36b (0.296 g, 0.69 mmol, 69%) as a white powder.

[0502]

[0503] 1 H NMR(CDCl3,400MHz)δ1.63-1.68(m,1H),2.17-2.26(m,1H),2.52-2.68(m,4H),4.75(s,2H),7.3 0(d,J=8.1Hz,2H),7.58(d,J=8.1Hz,2H),7.88(dd,J1=8.3Hz,J2=1.8Hz,1H),7.95-7.98(m,2H); 13 C NMR (CDCl3, 100MHz) δ13.7,31.5,64.4,67.5,109.9,114.9,121.9(q,J=272.6Hz),127.1(q,J= 4.7Hz), 128.3, 130.0, 132.2, 133.3, 133.4 (q, J = 33.2Hz), 134.2, 137.2, 142.9, 174.9, 179.9.

[0504] Example 37

[0505] 4-[5-(4-Formylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethyl-benzonitrile, 37a, [RD114]

[0506] To a mixture of 36b (0.303 g, 0.7 mmol) and Dess-Martin periodinane (0.417 g, 1 mmol) in dichloromethane (5 mL) was added pyridine (1.01 g, 1 mmol). The mixture was stirred at room temperature for 2 hours, after which diethyl ether (10 mL) was added to precipitate the reaction byproduct. After filtration and concentration under reduced pressure, the mixture was purified by chromatography (dichloromethane:acetone, 95:5) to afford 37a (0.24 g, 0.56 mmol, 80%) as a white powder.

[0507]

[0508] 1 H NMR(CDCl3,400MHz)δ1.62-1.73(m,1H),2.24-2.30(m,1H),2.50-2.58(m,2H),2.69-2.75(m,2H),7.53(d, J=8.1Hz,2H),7.85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.97-7.99(m,2H),8.11(d,J=8.1Hz,2H),10.12(s,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.7,67.5,110.2,114.8,121.9(q,J=272.6Hz),127.0(q,J=4.7Hz ), 129.1, 131.0, 131.2, 132.2, 133.3 (q, J = 33.2Hz), 135.3, 136.9, 140.5, 174.5, 179.8, 190.8.

[0509] Example 38

[0510] 4-{5-[4-(1-Hydroxyethyl)-phenyl]-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl}-2-trifluoromethyl-benzonitrile, 38a[RD116]

[0511] A mixture of 37a (0.043 g, 0.1 mmol) and dry THF (1 ml) in a flame-dried flash flask was placed under argon and cooled to -78°C. Then, methylmagnesium iodide (1.1 ml, 0.1 M) was added. The mixture was stirred at -78°C for 30 minutes and slowly warmed to room temperature. The mixture was washed with water (3 ml) and extracted with ethyl acetate (10 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to afford 38a (0.037 g, 0.082 mmol, 82%) as a white powder.

[0512]

[0513] 1 H NMR(CDCl3,400MHz)δ1.57(d,J=6.5Hz,3H),1.61-1.71(m,1H),2.09(d,J=3.2Hz,OH),2.16-2.28(m,1H),2.52-2.60(m,2H),2.63-2.69(m,2H ),5.00(dd,J1=6.5Hz,q,J2=3.1Hz,1H),7.29(d,J=8.3Hz,2H),7.60(d,J=8.2Hz,2H),7.85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.95-7.98(m,2H); 13 C NMR(CDCl3,100MHz)δ13.7,25.3,31.5,67.4,69.8,110.0,114.9,121.9(q,J=272.6Hz),127.0(q ,J=4.7Hz),127.1,129.9,132.2,133.4(q,J=33.2Hz),134.1,135.2,137.1,147.6,174.9,179.9.

[0514] Example 39

[0515] 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}-acrylate, ethyl ester, 39a[RD117]

[0516] A mixture of 37a (0.043 g, 0.1 mmol) and (ethoxycarbonylethylidene)triphenylphosphine (0.039 g, 0.12 mmol) in dichloromethane (2 ml) was stirred at room temperature for 10 hours. The mixture was concentrated and purified by chromatography (dichloromethane) to give 39a (0.048 g, 0.096 mmol, 96%) as a white powder.

[0517]

[0518] 1 H NMR(CDCl3,400MHz)δ1.35(t,J=7.1Hz,3H),1.66-1.70(m,1H),2.19-2.65(m,1H),2.51-2.69(m,2H),2.66-2.72(m,2H),4.28(q,J=7.1Hz,2H),6. 51(d,J=16.1Hz,1H),7.35(d,J=8.3Hz,2H),7.72(d,J=8.3Hz,2H),7.73(d,J=16.1Hz,1H),7.85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.96-7.98(m,2H); 13 C NMR (CDCl3, 100MHz) δ13.7,14.3,31.6,60.8,67.5,110.0,114.9,120.5,121.8(q,J=272.6Hz),127.0(q,J= 4.7Hz), 129.5, 130.5, 132.2, 133.4 (q, J = 33.2Hz), 135.2, 136.0, 136.5, 137.0, 142.7, 166.5, 174.7, 179.8.

[0519] Example 40

[0520] 4-{5-[4-(3-Hydroxypropenyl)-phenyl]-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl}-2-trifluoromethylbenzonitrile, 40a[RD120]

[0521] To a mixture of 39a (0.05 g, 0.1 mmol) in dichloromethane (2 ml) was added a solution of diisobutylaluminum hydride in THF (0.11 ml, 1 M, 0.11 mmol) at -78°C. The mixture was stirred at -78°C for 3 hours. After warming to room temperature, the mixture was washed with an aqueous solution of sodium thiosulfate and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to afford 40a (0.040 g, 0.089 mmol, 89%) as a white powder.

[0522]

[0523] 1 H NMR(CDCl3,400MHz)δ1.57-1.68(m,1H),2.17-2.39(m,1H),2.55-2.61(m,2H),2.61-2.67(m,2H),4.39(d,J=4.7Hz,2H),6.47(dt,J1=16.0Hz ,J2=5.3Hz,1H),6.70(d,J=16.0Hz,1H),7.29(d,J=8.3Hz,2H),7.59(d,J=8.3Hz,2H),7.85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.96-7.98(m,2H); 13 C NMR (CDCl3, 100MHz) δ13.7,31.5,63.4,67.4,110.0,114.8,120.5,121.8(q,J=272.6Hz),127.0(q,J= 4.7Hz), 127.9, 129.2, 130.1, 131.1, 132.1, 133.4 (q, J = 33.2Hz), 135.2, 137.1, 138.4, 174.8, 179.9.

[0524] Example 41

[0525] 41-1) 3-[4-(1-Cyanocyclobutylamino)-phenyl]-propionic acid, 41a(41-1)

[0526] Trimethylsilyl cyanide (0.4 g, 4 mmol) was added dropwise to a mixture of 3-(4-aminophenyl)-propionic acid (0.33 g, 2 mmol), cyclobutanone (0.35 g, 5 mmol), and sodium sulfate (1 g) in 1,4-dioxane (5 ml). The mixture was stirred for 15 hours. After filtering to remove the sodium sulfate, the mixture was concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane:acetone, 50:50) to give 41a (0.472 g, 1.93 mmol, 97%) as a light yellow solid.

[0527] 41-2) 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}-propionic acid methyl ester, 41b(41-2)[RD128]

[0528] A mixture of 1a (0.661 g, 2.9 mmol) and 41a (0.472 g, 1.93 mmol) in dry DMF (2 ml) was stirred at room temperature for 15 hours. To this mixture was added methanol (10 ml) and aqueous HCl (5 ml, 2 M). This second mixture was refluxed for 3 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 41b (0.582 g, 1.19 mmol, 62%) as a white powder.

[0529]

[0530] 1 H NMR(CDCl3,400MHz)δ1.60-1.70(m,1H),2.14-2.26(m,1H),2.51-2.56(m,2H),2.58-2.67(m,2H),2.71(t,J=7.8Hz,2H),3.05(t,J=7.8Hz,2H ),3.69(s,3H),7.23(d,J=8.2Hz,2H),7.41(d,J=8.2Hz,2H),7.85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.95(d,J=8.3Hz,1H),7.98(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,30.5,31.4,35.1,51.8,67.5,109.9,114.9,121.9(q,J=272.7Hz),127.1(q, J=4.7Hz),129.9,130.0,133.2,132.3,133.3(q,J=33.2Hz),135.7,137.2,142.5,173.1,174.9,179.9.

[0531] 41-3) 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-propionic acid, 41c(41-3)[RD132]

[0532] A mixture of 41b (0.487 g, 1 mmol) in methanol (10 ml) and sodium hydroxide solution (10 ml, 2 M) was stirred at room temperature for 5 hours. The methanol was evaporated. The residue was adjusted to pH 5 with aqueous HCl (2 M) and then extracted with ethyl acetate (3 x 50 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness to afford 41c (0.472 g, 0.99 mmol, 99%).

[0533] 41-4) 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-propionamide, 41d(41-4)[RD133]

[0534] To a suspension of 41c (0.094 g, 0.2 mmol) in THF (10 ml) was added thionyl chloride (0.019 ml, 0.26 mmol) at -5°C. The mixture was stirred at -5°C for 1 hour. Ammonia was then bubbled into the mixture. Excess ammonia was condensed by a reflux condenser at -78°C for 30 minutes and then evaporated. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 70:30) to afford 41d (0.09 g, 0.19 mmol, 95%) as an off-white powder.

[0535]

[0536] 1 H NMR (acetone-d6, 400MHz) δ1.52-160(m,1H),2.01-2.09(m,1H),2.49-2.58(m,4H),2.61-2.67(m,2H),2.98(t,J=7.5Hz,2H),6.20(bs,1H),6.7 8(bs,1H),7.31(d,J=8.2Hz,2H),7.44(d,J=8.2Hz,2H),8.03(dd,J1=8.3Hz,J2=1.8Hz,1H),8.15(d,J=1.8Hz,1H),8.22(d,J=8.3Hz,1H); 13 C NMR (acetone-d6, 100 MHz) δ 13.4, 30.7, 31.2, 36.4, 67.5, 109.0, 114.8, 122.5 (q, J = 271.5 Hz), 127.5 (q, J = 4.7 Hz), 129.5, 130.0, 131.8 (q, J = 32.5 Hz), 133.3, 133.8, 135.6, 138.4, 143.2, 171.6, 174.9, 178.0.

[0537] 41-5) 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-N-methyl-propionamide, 41e(41-5)[RD134]

[0538] To a suspension of 41c (0.094 g, 0.2 mmol) in THF (10 ml) was added thionyl chloride (0.019 ml, 0.26 mmol) at -5°C. The mixture was stirred at -5°C for 1 hour. Methylamine was bubbled through the mixture at -5°C for 30 minutes. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 75:25) to afford 41e (0.092 g, 0.19 mmol, 95%) as an off-white powder.

[0539]

[0540] 1 H NMR (acetone-d6, 400MHz) δ1.51-1.60(m,1H),2.01-2.11(m,1H),2.48-2.58(m,4H),2.61-2.67(m,2H),2.77(d,J=4.6Hz,3H),2.98(t,J=7.5Hz,2H) ,7.03(bs,NH),7.33(d,J=8.2Hz,2H),7.42(d,J=8.2Hz,2H),8.01(dd,J1=8.3Hz,J2=1.8Hz,1H),8.13(d,J=1.8Hz,1H),8.20(d,J=8.3Hz,1H); 13 C NMR (acetone-d6, 100 MHz) δ 13.4, 25.3, 30.0, 31.2, 37.0, 67.6, 109.0, 114.8, 122.5 (q, J = 271.5 Hz), 127.4 (q, J = 4.7 Hz), 129.5, 130.0, 131.9 (q, J = 32.5 Hz), 133.3, 133.8, 135.6, 138.4, 143.1, 171.7, 175.0, 178.0.

[0541] 41-6) 3-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-N-(2-hydroxyethyl)-propionamide, 41f(41-6)[RD135]

[0542] To a suspension of 41c (0.094 g, 0.2 mmol) in THF (10 ml) was added thionyl chloride (0.019 ml, 0.26 mmol) at -5°C. The mixture was stirred at -5°C for 1 hour. 2-Aminoethanol (0.0183 g, 0.03 mmol) was then added to the mixture at -5°C. After stirring for an additional 30 minutes, the mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 50:50) to afford 41f (0.093 g, 0.18 mmol, 90%) as an off-white powder.

[0543]

[0544] 1 H NMR (acetone-d6, 400MHz) δ1.51-161(m,1H),2.01-2.11(m,1H),2.49-2.66(m,6H),2.99 (t, J=7.5Hz, 2H), 3.27 (dd, J1=11.2Hz, J2=5.6Hz, 3H), 3.51 (dd, J1=11.2Hz, J2=5. 6Hz,2H),3.87(bs,OH),7.20(bs,NH),7.33(d,J=8.2Hz,2H),7.43(d,J=8.2Hz,2H) ,8.02(dd,J1=8.3Hz,J2=1.8Hz,1H),8.14(d,J=1.8Hz,1H),8.22(d,J=8.3Hz,1H); 13 CNMR (acetone-d6, 100 MHz) δ 13.4, 31.0, 31.2, 37.1, 42.0, 61.2, 67.6, 109.0, 114.8, 122.5 (q, J = 271.5 Hz), 127.4 (q, J = 4.7 Hz), 129.6, 130.0, 131.9 (q, J = 32.5 Hz), 133.3, 133.8, 135.6, 138.4, 143.0, 171.9, 175.0, 178.1.

[0545] 42-1) 4-[4-(1-Cyanocyclobutylamino)-phenyl]-butyric acid, 42a

[0546] Trimethylsilyl cyanide (0.50 g, 5 mmol) was added dropwise to a solution of 4-(4-aminophenyl)-butyric acid (0.537 g, 3 mmol), cyclobutanone (0.35 g, 5 mmol) and sodium sulfate (1 g) in 1,4-dichlorobenzene. The mixture was added to a mixture of 4-[4-(2-[ ...

[0547] 42-2) 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}-butyric acid methyl ester, 42b[RD129]

[0548] A mixture of 1a (0.547 g, 2.4 mmol) and 42a (0.342 g, 1.5 mmol) in dry DMF (2 ml) was stirred at room temperature for 15 hours. To this mixture was added methanol (10 ml) and aqueous HCl (5 ml, 2 M). This second mixture was refluxed for 3 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane) to afford 42b (0.594 g, 1.18 mmol, 79%) as a white powder.

[0549]

[0550] 1 H NMR(CDCl3,400MHz)δ1.60-1.70(m,1H),1.98-2.07(m,2H),2.14-2.26(m,1 H),2.40(t,J=7.4Hz,2H),2.52-2.60(m,2H),2.62-2.68(m,2H),2.74(t,J= 7.4Hz,2H),3.68(s,3H),7.22(d,J=8.2Hz,2H),7.38(d,J=8.2Hz,2H),7.86 (dd, J1=8.3Hz, J2=1.8Hz, 1H), 7.95 (d, J=8.3Hz, 1H), 7.98 (d, J=1.8Hz, 1H); 13 C NMR (CDCl3, 100MHz) δ13.7, 26.1, 31.4, 33.5, 34.8, 51.7, 67.5, 109.9, 114.9, 121.9 (q, J = 272.7Hz), 127.1 (q,J=4.7Hz),129.7,130.1,132.3,133.0,133.3(q,J=33.2Hz),135.2,137.2,143.5,173.8,175.0,179.9.

[0551] 42-3) 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-butyric acid, 42c[RD141]

[0552] A mixture of 42b (0.501 g, 1 mmol) in methanol (10 ml) and sodium hydroxide solution (10 ml, 2 M) was stirred at room temperature for 5 hours. The methanol was evaporated. The residue was adjusted to pH 5 with aqueous HCl (2 M), and the mixture was extracted with ethyl acetate (3 x 50 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness to afford 42c (0.482 g, 0.99 mmol, 99%), the structure of which is shown in Formula 5.

[0553]

[0554] 1 H NMR(CDCl3,400MHz)δ1.60-1.70(m,1H),1.98-2.07(m,2H),2.14-2.26(m,1H),2.45(t,J=7.3Hz,2H),2.51-2.59(m,2H),2.62-2.68(m,2H),2 .77(t,J=7.3Hz,2H),7.23(d,J=8.1Hz,2H),7.40(d,J=8.1Hz,2H),7.85(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.3Hz,1H),7.97(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,25.9,31.4,33.4,34.7,67.5,109.9,114.9,121.9(q,J=272.6Hz),127.1(q, J=4.7Hz),129.8,130.1,132.3,133.0,133.4(q,J=33.1Hz),135.2,137.2,143.3,174.9,178.9,179.9.

[0555] 42-4) 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-butyramide, 42d[RD130]

[0556] To a mixture of 42c (0.097 g, 0.2 mmol) in THF (10 ml) was added thionyl chloride (0.019 ml, 0.26 mmol) at -5°C. The mixture was stirred at -5°C for 1 hour. Ammonia was then bubbled into the mixture. Excess ammonia was condensed using a reflux condenser at -78°C for 30 minutes and then evaporated. The mixture was filtered. The filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 70:30) to afford 42d (0.093 g, 0.19 mmol, 95%) as an off-white powder.

[0557]

[0558] 1 H NMR(CDCl3,400MHz)δ1.57-1.70(m,1H),2.00-2.08(m,2H),2.16-2.25(m,1H), 2.31(t,J=7.3Hz,2H),2.51-2.59(m,2H),2.62-2.68(m,2H),2.77(t,J=7.3Hz,2 H),5.56(bs,1H),5.65(bs,1H),7.22(d,J=8.2Hz,2H),7.39(d,J=8.2Hz,2H),7 .85(dd,J1=8.3Hz,J2=1.8Hz,1H),7.95(d,J=8.3Hz,1H),7.97(d,J=1.8Hz,1H); 13 C NMR (CDCl3, 100MHz) δ13.7, 26.5, 31.4, 34.8, 35.0, 67.5, 109.9, 114.9, 121.9 (q, J = 272.7Hz), 127.1 (q, J=4.7Hz),129.8,130.1,132.2,133.0,133.3(q,J=33.2Hz),135.2,137.2,143.5,173.8,174.9,179.9.

[0559] 42-5) 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-N-methyl-butyramide, 42e[RD131]

[0560] To a suspension of 42c (0.097 g, 0.2 mmol) in THF (10 ml) was added thionyl chloride (0.019 ml, 0.26 mmol) at -5°C. The mixture was stirred at -5°C for 1 hour. Methylamine was then bubbled into the mixture at -5°C for 30 minutes. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 75:25) to afford 42e (0.095 g, 0.19 mmol, 95%) as an off-white powder.

[0561]

[0562] 1 H NMR(CDCl3,400MHz)δ1.52-1.64(m,1H),1.94-2.01(m,2H),2.10-2.17(m,1H ),2.20(t,J=7.3Hz,2H),2.46-2.62(m,4H),2.69(t,J=7.3Hz,2H),2.73(d,J =4.7Hz,3H),6.09(bs,1H),7.16(d,J=8.2Hz,2H),7.33(d,J=8.2Hz,2H),7.8 2(dd,J1=8.3Hz,J2=1.8Hz,1H),7.91(d,J=8.3Hz,1H),7.94(d,J=1.8Hz,1H); 13 C NMR (CDCl3, 100MHz) δ13.7, 26.2, 26.8, 31.4, 35.0, 35.7, 67.5, 109.7, 114.9, 121.9 (q, J = 272.7Hz), 127.1 (q, J = 4.7Hz), 129.7, 130.0, 132.3, 133.8, 133.3 (q, J = 33.2Hz), 135.2, 137.3, 143.7, 173.3, 174.9, 179.8.

[0563] 42-6) N-(4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}-butyryl)-methanesulfonamide, 42f[RD157]

[0564] A mixture of 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}butanoic acid (42c) (0.049 g, 0.1 mmol), 2,4,6-trichlorobenzoyl chloride (0.244 g, 1 mmol), 4-dimethylaminopyridine (0.122 g, 1 mmol) and methanesulfonamide (0.019 g, 0.2 mmol) in dichloromethane was stirred at room temperature for 20 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 80:20) to give N-(4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}-butyryl)-methanesulfonamide (42f) [RD157] (0.053 g, 0.094 mmol, 94%), the structure of which is shown in Formula 8, as a white powder.

[0565]

[0566] 1 H NMR (acetone-d6, 400MHz) δ1.51-160(m,1H),1.96-2.11(m,3H),2.49(t,J=7.3Hz,2H),2.51-2.57(m,2H),2.61-2.67(m,2H),2.75(t,J=7.5Hz,2H),2. 94(bs,1H),3.24(s,3H),7.33(d,J=8.3Hz,2H),7.43(d,J=8.2Hz,2H),8.02(dd,J=8.3,1.6Hz,1H),8.02(d,J=1.6Hz,1H),8.21(d,J=8.3Hz,1H); 13 CNMR (acetone-d6, 100 MHz) δ 13.4, 25.8, 31.2, 34.3, 35.2, 40.6, 67.6, 109.0, 114.8, 122.5 (q, J = 271.5 Hz), 127.5 (q, J = 4.9 Hz), 129.6, 130.1, 131.9 (q, J = 33.6 Hz), 133.3, 133.9, 135.6, 138.4, 143.1, 171.9, 175.0, 180.5.

[0567] 42-7) N-Methyl-4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-6,8-dioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}butanamide, 42 g [RD158]

[0568] Hydrogen peroxide (30%, 0.4%) was added dropwise to a mixture of N-methyl-4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}butanamide (42e) (0.032 g, 0.064 mmol) in glacial acetic acid (0.5 ml). The mixture was stirred at room temperature for 5 hours, then washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated and purified by chromatography (dichloromethane:acetone, 80:20) to give N-methyl-4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-6,8-dioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}butanamide (42 g) [RD158] (0.029 g, 0.06 mmol, 94%), the structure of which is shown in Formula 9, as a white powder.

[0569]

[0570] 1 H NMR(CDCl3,400MHz)δ1.63-1.71(m,1H),1.93-2.04(m,2H),2.18-2.27(m, 3H),2.44-2.53(m,2H),2.57-2.65(m,2H),2.70(t,J=7.3Hz,2H),2.79(d, J=4.8Hz,3H),5.79(bs,1H),7.21(d,J=8.2Hz,2H),7.34(d,J=8.2Hz,2H), 7.92(d,J=8.4Hz,1H), 8.03(dd,J=8.3,1.8Hz,1H), 8.18(d,J=1.8Hz,1H).

[0571] Example 43

[0572] 43-1) tert-Butyl 4-(4-aminophenyl)-piperazine-1-carboxylate, 43a

[0573] A mixture of 4-iodoaniline (0.654 g, 3 mmol), tert-butyl piperazine-1-carboxylate (0.67 g, 3.6 mmol), potassium phosphate (1.272 g, 6 mmol), ethylene glycol (0.33 ml), and copper iodide (0.03 g, 0.15 mmol) in 2-propanol (3 ml) was placed in a sealed tube under nitrogen and heated to 80°C for 30 hours. After cooling to room temperature, the mixture was washed with water (50 ml) and extracted with ethyl acetate (100 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 70:30) to afford 43a (0.36 g, 1.3 mmol, 43%) as a yellow powder.

[0574] 43-2) tert-Butyl 4-[4-(1-cyanocyclobutylamino)phenyl]-piperazine-1-carboxylate, 43b

[0575] Trimethylsilyl cyanide (0.3 g, 3 mmol) was added dropwise to a mixture of 43a (0.415 g, 1.5 mmol), cyclobutanone (0.21 g, 3 mmol), and sodium sulfate (1 g) in dichloromethane (5 ml). The mixture was stirred for 15 hours. After filtering to remove the sodium sulfate, the mixture was concentrated under vacuum to give a brown liquid, which was purified by chromatography (dichloromethane:acetone, 75:25) to give 43b (0.448 g, 1.26 mmol, 84%) as a yellow solid.

[0576] 43-3) 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-imino-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}-piperazine-1-carboxylic acid tert-butyl ester, 43c[RD139]

[0577] and 4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-(4-cyano-3-trifluoromethyl-phenylthiocarbamoylimino)-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-phenyl}-piperazine-1-carboxylic acid tert-butyl ester, 43d[RD140]

[0578] A mixture of 1a (0.228 g, 1 mmol) and 43b (0.472 g, 0.63 mmol) in dry DMF (1 ml) was stirred at room temperature for 20 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 90:10) to afford 43c (0.173 g, 0.296 mmol, 47%), whose structure is shown in Formula 10, as an off-white powder, and 43d (0.169 g, 0.21 mmol, 33%), whose structure is shown in Formula 11, as a yellow powder.

[0579]

[0580] 1 H NMR(CDCl3,400MHz)δ1.48,(s,9H),1.57-1.67(m,1H),2.01-2.09(m,1H),2.59-2.70(m,4H),3.25(t,J=5.1Hz,4H),3.59 (t,J=4.9Hz,4H),7.02(d,J=8.9Hz,2H),7.20(d,J=8.9Hz,2H),7.81(d,J=7.4Hz,1H),7.93(s,1H),7.97(d,J=8.1Hz,1H).

[0581]

[0582] 1 H NMR(CDCl3,400MHz)δ1.48,(s,9H),1.57-1.64(m,1H),2.01-2.10(m,1H),2.60-2.89(m,4H),3.24(t,J=5.1Hz,4 H), 3.57 (t, J = 4.9Hz, 4H), 7.02 (d, J = 8.9Hz, 2H), 7.20 (d, J = 8.9Hz, 2H), 7.54-7.98 (m, 4H), 7.97 (d, J = 8.1Hz, 1H).

[0583] 43-4) 4-[8-Oxo-5-(4-piperazin-1-yl-phenyl)-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethylbenzonitrile, 43e[RD137]

[0584] A mixture of 43c (0.117 g, 0.2 mmol), methanol (5 ml), and aqueous HCl (2 ml, 2 M) was refluxed for 2 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 50:50 followed by methanol:acetone, 50:50) to afford 43e (0.089 g, 0.184 mmol, 92%) as a white powder.

[0585]

[0586] 1 H NMR(CD3OD,400MHz)δ1.51-1.61(m,1H),2.01-2.11(m,1H),2.48-2.59(m,4H),2.90-2.97(m,4H),3.25-3.30(m,4H),7.03 (d,J=8.9Hz,2H),7.16(d,J=8.9Hz,2H),7.86(dd,J1=8.3Hz,J2=1.8Hz,1H),8.02(d,J=8.3Hz,1H),8.07(d,J=1.8Hz,1H); 13C NMR(CD3OD,100MHz)δ13.2,30.9,45.1,48.9,67.5,108.9,114.8,115.9,122.3(q,J=271.7Hz),1 26.4, 127.3 (q, J = 4.7Hz), 130.4, 132.2 (q, J = 33.2Hz), 133.0, 135.4, 138.1, 152.1, 175.4, 180.4.

[0587] 43-5) 4-{5-[4-(4-Methanesulfonylpiperazin-1-yl)-phenyl]-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl}-2-trifluoromethylbenzonitrile, 43f[RD138]

[0588] A mixture of 43e (0.049 g, 0.1 mmol), methanesulfonyl chloride (0.012 ml, 0.15 mmol) and triethylamine (0.15 ml) in dichloromethane was stirred at room temperature for 5 hours. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 95:5) to afford 43f (0.042 g, 0.074 mmol, 74%) as a white powder.

[0589]

[0590] 1 H NMR(CDCl3,400MHz)δ1.62-1.70(m,1H),2.14-2.23(m,1H),2.51-2.58(m,2H),2.61-2.67(m,2H),2.84(s,3H),3.39(s,8H),7 .05(d,J=8.9Hz,2H),7.20(d,J=8.9Hz,2H),7.84(dd,J1=8.3Hz,J2=1.8Hz,1H),7.95(d,J=8.3Hz,1H),7.97(d,J=1.8Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.4,34.6,45.7,48.4,67.5,109.8,114.9,117.0,121.9(q,J=272.7Hz) ,126.8,127.1(q,J=4.7Hz),130.7,132.3,133.4(q,J=33.2Hz),135.2,137.3,151.1,175.0,180.2.

[0591] Example 44

[0592] 44-1) 3-{4-[7-(4-cyano-3-trifluoromethyl-phenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-acrylic acid, 44a

[0593] A mixture of 39a (0.025 g, 0.05 mmol) in methanol (2 ml) and sodium hydroxide solution (2 ml, 2 M) was stirred at room temperature for 5 hours. The methanol was evaporated. The residue was adjusted to pH 5 with aqueous HCl (2 M) and then extracted with ethyl acetate (3 x 50 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness to afford 44a (0.02 g, 0.042 mmol, 85%).

[0594] 44-2) 3-{4-[7-(4-cyano-3-trifluoromethyl-phenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-phenyl}-acrylamide, 44b[RD119]

[0595] To a suspension of 44b (0.02 g, 0.042 mmol) in THF (1 ml) was added thionyl chloride (0.007 ml, 0.1 mmol) at -5°C. The mixture was stirred for 1 hour. Ammonia was then bubbled into the mixture. Excess ammonia was condensed using a reflux condenser at -78°C for 30 minutes and then evaporated. The mixture was filtered. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 70:30) to afford 44b (0.014 g, 0.03 mmol, 71%) as an off-white powder.

[0596]

[0597] 1 H NMR(DMSO-d6,400MHz)δ1.49-1.52(m,1H),1.88-1.93(m,1H),2.37-2.46(m,2H),2.57-2.62(m,2H),6.66(d,J=15.9Hz,1H),7.16(bs,1H),7 .43(d,J=8.3Hz,2H),7.47(d,J=15.9Hz,1H),7.58(bs,1H),8.03(dd,J1=8.3Hz,J2=1.8Hz,1H),8.23(d,J=1.8Hz,1H),8.34(d,J=8.3Hz,1H).

[0598] Example 45 [RD145]

[0599] Trimethylsilyl cyanide (0.4 g, 4 mmol) was added dropwise to a mixture of 4-methanesulfonylaniline hydrochloride (0.415 g, 2 mmol), cyclobutanone (0.28 g, 4 mmol), and sodium sulfate (1 g) in DMF (3 ml). The mixture was stirred at 120°C for 15 hours. After filtering to remove the sodium sulfate, the filtrate was washed with brine and extracted with ethyl acetate. The organic layer was concentrated and purified by chromatography (dichloromethane:acetone, 90:10) to afford 1-(4-methanesulfonylphenylamino)cyclobutanecarbonitrile (45a) (0.116 g, 0.44 mmol, 22%) as a pale yellow solid. 4-methanesulfonylaniline (0.201 g, 1.17 mmol, 59%) was recovered.

[0600] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.0.141 g, 0.62 mmol) and 1-(4-methylsulfonylphenylamino)cyclobutanecarbonitrile (45a) (0.11 g, 0.42 mmol) in dry DMF (2 ml) was stirred at room temperature for 3 days. Methanol (10 ml) and 2N aqueous HCl (5 ml) were added to this mixture. This second mixture was refluxed for 3 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 97:3) to afford 4-[5-(4-methylsulfonylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethylbenzonitrile (45b) [RD145] (0.031 g, 0.065 mmol, 15%), the structure of which is shown in Formula 14, as a white powder.

[0601]

[0602] 1 H NMR(CDCl3,400MHz)δ1.63-1.72(m,1H),2.21-2.28(m,1H),2.46-2.54(m,2H),2.68.2.74(m,2H),3.16(s,3H),7.5 7(d,J=8.3Hz,2H),7.85(dd,J=8.3,1.8Hz,1H),7.97(d,J=1.8Hz,1H),7.98(d,J=8.3Hz,1H),8.17(d,J=8.3Hz,2H); 13C NMR(CDCl3,100MHz)δ13.6,31.8,44.4,67.5,110.2,114.8,122.4(q,J=271.5Hz),127.0(q,J= 4.9Hz), 129.4, 131.4, 132.1, 133.6 (q, J = 33.3Hz), 135.3, 136.8, 140.3, 141.8, 174.4, 179.9.

[0603] Example 46

[0604] Trimethylsilyl cyanide (0.69 g, 7 mmol) was added dropwise to a mixture of 4-aminophenylacetic acid (0.755 g, 5 mmol) and cyclobutanone (0.49 g, 7 mmol) in distilled water. The mixture was added to a mixture of 4-nitropropane (20 ml). The mixture was stirred at 80°C for 8 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 60:40) to give [4-(1-cyanocyclobutylamino)phenyl]acetic acid (46a) (1.138 g, 4.95 mmol, 99%) as a white solid.

[0605] 46-1)RD146

[0606] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.638 g, 2.8 mmol) and [4-(1-cyanocyclobutylamino)phenyl]acetic acid (46a) (0.46 g, 2.0 mmol) in DMF (5 ml) was stirred at room temperature for 15 hours. Methanol (20 ml) and 2N aqueous HCl (10 ml) were added to this mixture. This second mixture was refluxed for 1 hour. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 50 ml). The organic layer was dried over magnesium sulfate, concentrated and purified by chromatography (neat dichloromethane then dichloromethane:acetone, 95:5) to give {4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}acetate methyl ester (46b) [RD146] (0.532 g, 1.124 mmol, 56%), the structure of which is shown in Formula 15, as a white powder.

[0607]

[0608] 1H NMR(CDCl3,400MHz)δ1.60-1.69(m,1H),2.15-2.25(m,1H),2.50-2.58(m,2H),2.61-2.66(m,2H),3.72(bs,5H),7.2 7(d,J=8.3Hz,2H),7.50(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.8Hz,1H),7.94(d,J=8.2Hz,1H),7.97(d,J=1.6Hz,1H); 13 C NMR (CDCl3, 100MHz) δ13.7,31.4,44.7,52.3,67.4,109.9,114.9,122.0(q,J=272.5Hz),127.0(q,J= 4.9Hz), 130.0, 131.1, 132.3, 133.0 (q, J = 33.3Hz), 134.1, 135.2, 135.9, 137.2, 171.4, 174.9, 179.9.

[0609] 46-2)RD147

[0610] A mixture of methyl {4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]oct-5-yl]phenyl}acetate (46b) (0.095 g, 0.2 mmol) and a 2M sodium hydroxide solution (1 ml) in methanol (2 ml) was stirred at room temperature for 2 hours. The methanol was evaporated. The residue was adjusted to pH 5 with 2M HCl, and the mixture was extracted with ethyl acetate (3 x 10 ml). The organic layer was dried over magnesium sulfate and concentrated to dryness to give {4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]oct-5-yl]phenyl}acetic acid (46c) [RD147] (0.087 g, 0.19 mmol, 95%), the structure of which is shown in Formula 16.

[0611]

[0612] 1 H NMR (CDCl3, 400MHz) δ1.60-1.69(m,1H),2.15-2.25(m,1H),2.50-2.64(m,4H),3.73(s,2H),7.26(d,J=8. 3Hz,2H),7.51(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.2Hz,1H),7.96(d,J=1.6Hz,1H); 13CNMR(CDCl3,100MHz)δ13.7,31.4,40.2,40.8,67.4,109.9,114.9,122.0(q,J=272.5Hz),127.0(q,J= 4.9Hz), 129.9, 131.2, 132.3, 133.3 (q, J = 33.3Hz), 133.9, 135.2, 136.1, 137.2, 174.1, 174.9, 179.9.

[0613] 46-3)RD148

[0614] Thionyl chloride (0.238 g, 2 mmol) was added dropwise to a mixture of {4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}acetic acid (46c) (0.357 g, 0.777 mmol) in THF (5 ml) cooled to 0°C. The mixture was stirred at room temperature for 1 hour, and then ammonia was bubbled into the mixture. Excess ammonia was condensed at -78°C using a reflux condenser for 30 minutes and then evaporated. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 70:30) to give 2-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}acetamide (46d) [RD148] (0.345 g, 0.75 mmol, 97%), the structure of which is shown in Formula 17, as an off-white powder.

[0615]

[0616] 1 H NMR(CDCl3,400MHz)δ1.62-1.66(m,1H),2.18.2.23(m,1H),2.49-2.55(m,2H),2.61-2.66(m,2H),3.63(s,2H),5.91(bs,1H),6.1 0(bs,1H),7.27(d,J=8.1Hz,2H),7.50(d,J=8.1Hz,2H),7.83(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.2Hz,1H),7.96(d,J=1.6Hz,1H); 13C NMR(CDCl3,100MHz)δ13.7,31.5,42.5,67.4,109.9,114.9,121.9(q,J=272.4Hz),127.1(q,J=4.9 Hz), 130.2, 131.1, 132.2, 133.3 (q, J = 33.3Hz), 134.1, 135.2, 136.8, 137.2, 172.8, 174.8, 180.0.

[0617] 46-4)RD149

[0618] Thionyl chloride (0.238 g, 2 mmol) was added dropwise to a mixture of {4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}acetic acid (46c) (0.357 g, 0.777 mmol) in THF (5 ml) cooled to 0° C. The mixture was stirred at room temperature for 1 hour, and then methylamine (0.5 ml) was added to the mixture. The mixture was stirred for another 2 hours. The mixture was filtered, and the filtrate was concentrated and purified by chromatography (dichloromethane:acetone, 80:20) to give N-methyl-2-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}acetamide (46e) [RD149] (0.348 g, 0.738 mmol, 95%), the structure of which is shown in Formula 18, as an off-white powder.

[0619]

[0620] 1 H NMR(CDCl3,400MHz)δ1.61-1.70(m,1H),2.17-2.31(m,1H),2.50-2.56(m,2H),2.61-2.68(m,2H),2.82(d,J=4.8Hz,3H),3.62( s,2H),7.27(d,J=8.3Hz,2H),7.50(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.2Hz,1H),7.96(d,J=1.6Hz,1H); 13CNMR(CDCl3,100MHz)δ13.7,26.6,31.5,43.1,67.4,110.0,114.9,122.0(q,J=272.5Hz),127.1(q,J= 4.9Hz), 130.2, 131.0, 132.2, 133.3 (q, J = 33.3Hz), 134.1, 135.2, 137.0, 137.1, 170.1, 174.8, 179.9.

[0621] Example 47

[0622] N-{4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]phenyl}methanesulfonamide (47a) [RD150]

[0623] A mixture of 4-[3-(4-aminophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile (2d) (0.02 g, 0.05 mmol), methanesulfonyl chloride (0.009 g, 0.075 mmol) and pyridine (0.006 g, 0.075 mmol) in dichloromethane (1 ml) was stirred at room temperature for 15 hours. The mixture was washed with water (2 ml) and extracted with ethyl acetate (5 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (HPLC, alumina column) to obtain N-{4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]phenyl}methanesulfonamide (47a) [RD150] (0.009 g, 0.018 mmol, 36%), whose structure is shown in Formula 2, as a white powder.

[0624]

[0625] 1 H NMR(DMSO-d6,400MHz)δ1.46(s,6H),3.07(s,3H),7.32(s,4H),8.05(dd,J=8 .2,1.2Hz,1H),8.26(d,J=1.2Hz,1H),8.35(d,J=8.2Hz,1H),10.08(bs,1H); 13C NMR(DMSO-d6,100MHz)δ23.3,40.4,66.7,109.0,115.5,119.9,122.6(q,J=272.2Hz),128.5 (q, J=4.7Hz), 130.8, 131.2, 131.5 (q, J= 32.3Hz), 134.5, 136.6, 138.6, 139.5, 175.4, 180.4.

[0626] Example 48

[0627] N-{4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]phenyl}acetamide, 48a, [RD151]

[0628] A mixture of 4-[3-(4-aminophenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethylbenzonitrile (2d) [RD9] (0.008 g, 0.02 mmol), acetyl chloride (0.004 g, 0.03 mmol), and triethylamine (0.003 g, 0.03 mmol) in dichloromethane (1 ml) was stirred at 0°C for 2 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 90:10) to give N-{4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl]phenyl}acetamide, 48a, [RD151] (0.007 g, 0.016 mmol, 80%), the structure of which is shown in Formula 3, as a white powder.

[0629]

[0630] 1 H NMR (CDCl3, 400MHz) δ1.58 (s, 6H), 2.21 (s, 3H), 7.24 (d, J = 8.6Hz, 2H), 7.48 (bs, 1H), 7.69 ( d,J=8.6Hz,2H),7.83(dd,J=8.2,1.9Hz,1H),7.96(d,J=1.2Hz,1H),7.97(d,J=8.2Hz,1H); 13 C NMR(CDCl3,100MHz)δ23.6,53.4,66.4,110.0,114.8,120.7,122.6(q,J=272.2Hz),127.1(q,J = 4.7Hz), 129.1, 130.2, 132.2, 133.5 (q, J = 32.3Hz), 135.2, 137.1, 139.2, 168.1, 175.0, 180.0.

[0631] Example 49

[0632] Concentrated sulfuric acid was slowly added to a solution of 4-aminobenzoic acid (4 g, 29.2 mmol) in methanol cooled to 0°C. After the addition was complete, the mixture was stirred at room temperature for 5 hours. The mixture was washed with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated under vacuum to afford methyl 4-aminobenzoate (49a) (4.22 g, 27.9 mmol, 96%) as an off-white solid.

[0633] A mixture of methyl 4-aminobenzoate (0.32 g, 2.12 mmol), acetone cyanohydrin (3 ml), and sodium sulfate (1 g) was refluxed for 15 hours. After filtering to remove the sodium sulfate, the filtrate was washed with brine and extracted with ethyl acetate. The organic layer was concentrated and purified by chromatography (dichloromethane:acetone, 60:40) to afford methyl 4-[(cyanodimethylmethyl)-amino]-benzoate (49b) (0.398 g, 1.95 mmol, 92%) as a white solid.

[0634] 49-1)RD152

[0635] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.228 g, 1 mmol) and 4-[(cyanodimethylmethyl)-amino]-benzoic acid methyl ester (49b) (0.14 g, 0.64 mmol) in DMF (2 ml) was heated at 60°C under microwave irradiation for 12 hours. Methanol (6 ml) and 2N aqueous HCl (2 ml) were added to this mixture. This second mixture was refluxed for 4 hours. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane; dichloromethane:acetone, 75:25) to give methyl 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]benzoate (49c) [RD152] (0.18 g, 0.4 mmol, 63%), the structure of which is shown in Formula 19, as a white powder.

[0636]

[0637] 1H NMR (CDCl3, 400MHz) δ1.60 (s, 6H), 3.95 (s, 3H), 7.40 (d, J = 8.6Hz, 2H), 7.84 (dd, J = 8 .2,1.9Hz,1H),7.96(d,J=1.2Hz,1H),7.97(d,J=8.2Hz,1H),8.21(d,J=8.6Hz,2H); 13 C NMR(CDCl3,100MHz)δ23.8,52.6,66.6,110.3,114.8,121.9(q,J=272.7Hz),127.1(q,J=4.7Hz ), 129.8, 131.2, 131.4, 132.2, 133.5 (q, J = 32.3Hz), 135.3, 137.0, 139.2, 165.9, 174.7, 179.7.

[0638] 49-2)RD153

[0639] A mixture of methyl 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]benzoate (49c) (0.02 g, 0.0435 mmol) and methylamine (2 ml, distilled from a 40% aqueous solution thereof) was kept at -20°C for 15 hours. After evaporation of the methylamine, the mixture was purified by chromatography (dichloromethane:acetone, 80:20) to give 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]-N-methylbenzamide (49d) [RD153] (0.01 g, 0.0224, 51%), the structure of which is shown in Formula 20. The ester 4-[3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]benzoic acid methyl ester (49c) was recovered (0.08 g, 0.0179 mmol, 41%).

[0640]

[0641] 1 H NMR (acetone-d6, 400MHz) δ1.60 (s, 6H), 2.90 (d, J = 4.6Hz, 3H), 7.48 (d, J = 8.6Hz, 2H), 7.80 (bs, 1H), 7 .99(d,J=8.6Hz,2H),8.06(dd,J=8.2,1.8Hz,1H),8.18(d,J=1.8Hz,1H),8.25(d,J=8.2Hz,1H); 13C NMR (acetone-d6, 100 MHz) δ 23.8, 54.0, 66.5, 110.3, 114.8, 121.9 (q, J = 272.7 Hz), 127.1 (q, J = 4.7 Hz), 128.2, 129.9, 133.5 (q, J = 32.3 Hz), 135.7, 135.8, 138.2, 138.3, 139.2, 166.0, 174.9, 179.7.

[0642] Example 50

[0643] 50-1)RD154

[0644] A mixture of 4-[8-(4-hydroxymethylphenyl)-5-oxo-7-thioxo-6-azaspiro[3.4]octan-6-yl]-2-trifluoromethyl-benzonitrile (36b) (0.086 g, 0.2 mmol) and methanesulfonic anhydride (0.07 g, 0.4 mmol) in dichloromethane (1 ml) was stirred at room temperature for 15 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 98:2) to give methanesulfonic acid 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]phenylmethyl ester (50a) [RD154] (0.089 g, 0.175 mmol, 88%), the structure of which is shown in Formula 22, as a white powder.

[0645]

[0646] 1 H NMR(CDCl3,400MHz)δ1.63-1.70(m,1H),2.17-2.31(m,1H),2.48-2.57(m,2H),2.64-2.70(m,2H),3.04(s,3H),5.30(s,2H) ), 7.37 (d, J = 8.3Hz, 2H), 7.62 (d, J = 8.3Hz, 2H), 7.84 (dd, J = 8.3, 1.8Hz, 1H), 7.97 (d, J = 8.2Hz, 1H), 7.98 (d, J = 1.6Hz, 1H).

[0647] 50-2)RD155

[0648] Methylamine (0.5 ml) was bubbled into a mixture of 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]phenylmethyl methanesulfonate (50a) (0.059 g, 0.115 mmol) in THF (3 ml) cooled to -78°C. After reacting at -78°C for 1 hour, the mixture was concentrated and purified by chromatography (dichloromethane:acetone, 95:5; methanol) to give 4-[5-(4-methylaminomethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethylbenzonitrile (50b) [RD155] (0.042 g, 0.095 mmol, 82%), the structure of which is shown in Formula 23, as a white powder.

[0649]

[0650] 1 H NMR(CDCl3,400MHz)δ1.57-1.70(m,1H),2.16-2.24(m,1H),2.52(s,3H),2.53-2.57(m,2H),2.60-2.68(m,2H),3.85(s,2H) ),7.27(d,J=8.3Hz,2H),7.55(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.2Hz,1H),7.97(d,J=1.6Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.5,36.4,55.6,67.4,110.0,114.9,122.0(q,J=272.5Hz),127.0(q ,J=4.9Hz),129.1,129.6,129.8,132.2,133.3(q,J=33.3Hz),133.7,135.2,142.4,174.8,179.9.

[0651] 50-3)RD156

[0652] A mixture of 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]phenylmethyl methanesulfonate (50a) (0.02 g, 0.039 mmol) and dimethylamine (0.5 ml; distilled from a 40% aqueous solution thereof) in THF (1 ml) was stirred at -78°C for 2 hours. The mixture was concentrated and purified by chromatography (dichloromethane:acetone, 95:5; acetone) to give 4-[5-(4-dimethylaminomethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethylbenzonitrile (50c) [RD156] (0.017 g, 0.037 mmol, 95%), the structure of which is shown in Formula 24, as a white powder.

[0653]

[0654] 1 H NMR(CDCl3,400MHz)δ1.57-1.70(m,1H),2.16-2.24(m,1H),2.32(s,6H),2.55-2.60(m,2H),2.63-2.69(m,2H),3.53(s,2H) ),7.27(d,J=8.3Hz,2H),7.55(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.8Hz,1H),7.95(d,J=8.2Hz,1H),7.97(d,J=1.6Hz,1H); 13 C NMR(CDCl3,100MHz)δ13.7,31.5,45.5,63.7,67.4,110.0,114.9,122.0(q,J=272.5Hz),127.0(q ,J=4.9Hz),129.1,129.6,129.8,132.2,133.3(q,J=33.3Hz),133.7,135.2,142.4,174.8,179.9.

[0655] Example 51

[0656] Sodium cyanide (0.245 g, 5 mmol) was added to a mixture of 4-aminobenzoic acid (0.274 g, 2 mmol) and cyclobutanone (0.21 g, 3 mmol) in 90% acetic acid (4.5 ml). The reaction mixture was stirred at room temperature for 15 hours. The mixture was washed with aqueous HCl (pH 2) and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated to dryness under vacuum to give 4-(1-cyanocyclobutylamino)benzoic acid (51a) (0.426 g, 1.97 mmol, 99%) as a white solid.

[0657] 51-1) RD159 and RD160

[0658] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.51 g, 2.22 mmol) and 4-(1-cyanocyclobutylamino)benzoic acid (51a) (0.343 g, 1.59 mmol) in DMF (2 ml) was heated at 60°C under microwave irradiation and stirred for 16 hours. Methanol (10 ml) and 2M aqueous HCl (5 ml) were added to this mixture. This second mixture was refluxed for 12 hours. After cooling to room temperature, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (3 x 30 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to give 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-benzoic acid methyl ester (51b) [RD159] (0.09 g, 0.196 mmol, 12%), the structure of which is shown in Formula 25, as a white powder, and N-(3-cyano-4-trifluoromethylphenyl)-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]benzamide (51b') [RD160] (0.28 g, 0.45 mmol, 29%), the structure of which is shown in Formula 26, as a white powder.

[0659]

[0660] 1 H NMR(CDCl3,400MHz)δ1.67-1.71(m,1H),2.20-2.26(m,1H),2.49-2.57(m,2H),2.66-2.73(m,2H),3.96(s,3H),7.4 2(d,J=8.4Hz,2H),7.85(dd,J=8.3,1.7Hz,1H),7.97(d,J=8.3Hz,1H),7.98(d,J=1.7Hz,1H),8.26(d,J=8.3Hz,2H); 13 C NMR (CDCl3, 100MHz) δ13.7,31.6,52.6,67.5,110.1,114.8,121.8(q,J=272.7Hz),127.0(q,J=4.7 Hz), 130.2, 131.4, 131.5, 132.2, 133.4 (q, J = 33.2Hz), 135.2, 137.0, 139.2, 165.9, 174.6, 179.7.

[0661]

[0662]

[0663] 1 H NMR(CDCl3,400MHz)δ1.67-1.71(m,1H),2.18-2.26(m,1H),2.50-2.58(m,2H),2.68-2.74(m,2H),7.47(d,J=8.5Hz,2H),7.83(d,J=8.7H z,1H),7.84(dd,J=8.3,1.9Hz,1H),7.96(d,J=8.0Hz,1H),9.97(d,J=1.9Hz,1H),8.10-8.14(m,3H),8.21(d,J=1.9Hz,1H),8.88,(s,1H).

[0664] 51-2)RD161

[0665] A mixture of 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]-benzoic acid methyl ester (51b) (0.046 g, 0.1 mmol) and methylamine (1 ml, distilled from a 40% aqueous solution thereof) was kept at -20°C for 15 hours. After evaporation of the methylamine, the mixture was purified by chromatography (dichloromethane:acetone, 80:20) to give N-methyl-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl]benzamide (51c) [RD161] (0.041 g, 0.085, 84%), the structure of which is shown in Formula 27.

[0666]

[0667] 1 H NMR(CDCl3,400MHz)δ1.63-1.70(m,1H),2.18-2.26(m,1H),2.48-2.56(m,2H),2.65-2.71(m,2H),3.05 (d,J=4.8Hz,3H),6.32(bs,1H),7.39(d,J=8.3Hz,2H),7.84(dd,J=8.3,1.7Hz,1H),7.95-7.98(m,4H); 13C NMR(CDCl3,100MHz)δ13.6,27.0,31.6,67.4,110.3,114.8,121.8(q,J=272.7Hz),127.0(q,J=4.7 Hz), 128.7, 130.3, 132.1, 133.3 (q, J = 33.2Hz), 135.2, 136.3, 137.0, 137.8, 167.2, 174.6, 179.8.

[0668] Example 52 [RD162]

[0669] Thionyl chloride (2.38 g, 20 mmol) was slowly added to a solution of 2-fluoro-4-nitrobenzoic acid (2.97 g, 16 mmol) in DMF (50 ml) cooled to -5°C. The mixture was stirred at -5°C for another hour. Methylamine (0.62 g, 20 mmol; freshly prepared, distilled from a 40% aqueous solution) was added to the reaction mixture. This second mixture was stirred for another hour. Ethyl acetate (300 ml) was added to the mixture, which was washed with brine (3 x 150 ml). The organic layer was dried over magnesium sulfate and concentrated to afford N-methyl-2-fluoro-4-nitrobenzamide (52a) (2.89 g, 14.6 mmol, 91%) as a yellow solid.

[0670] 1 H NMR (acetone d6, 400 MHz) δ 3.05 (d, J = 4.3 Hz, 3H), 6.31 (dd, J = 13.5, 2.1 Hz, 1H), 6.40 (dd, J = 8.5, 2.1 Hz, 1H), 7.64 (dd, J = 8.6, 8.6 Hz, 1H).

[0671] A mixture of N-methyl-2-fluoro-4-nitrobenzamide (52a) (2.89 g, 14.6 mmol) and iron powder (5.04 g, 90 mmol) in ethyl acetate (40 ml) and acetic acid (40 ml) was refluxed for 1 hour. Solid particles were filtered off. The filtrate was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to afford N-methyl-2-fluoro-4-aminobenzamide (52b) (2.3 g, 13.7 mmol, 94%) as an off-white solid.

[0672] 1H NMR (acetone-d6, 400 MHz) δ 2.86 (d, J = 4.3 Hz, 3H), 5.50 (bs, 2H), 6.37 (dd, J1 = 14.7 Hz, J2 = 2.1 Hz, 1H), 6.50 (dd, J = 8.5, 2.1 Hz, 1H), 7.06 (bs, 1H), 7.68 (dd, J = 8.88.8 Hz, 1H); 13 C NMR (acetone-d6, 100 MHz) δ 25.8, 99.6 (d, J = 13.8 Hz), 109.2 (d, J = 12.8 Hz), 110.0 (d, J = 1.6 Hz), 132.5 (d, J = 4.8 Hz), 153.5 (d, J = 12.6 Hz), 162.2 (d, J = 242.5 Hz), 164.0 (d, J = 3.1 Hz).

[0673] Sodium cyanide (1.47 g, 30 mmol) was added to a mixture of N-methyl-2-fluoro-4-aminobenzamide (52b) (1.68 g, 10 mmol) and cyclobutanone (1.4 g, 20 mmol) in 90% acetic acid (20 ml). The reaction mixture was stirred at 80°C for 24 hours. The mixture was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated to dryness under vacuum. The solid was washed with a 50:50 mixture of diethyl ether and hexane (10 ml) to remove cyclobutanone cyanohydrin and filtered to give N-methyl-4-(1-cyanocyclobutylamino)-2-fluorobenzamide (52c) (2.19 g, 8.87 mmol, 89%).

[0674] 1 H NMR(CDCl3,400MHz)δ1.87-1.95(m,1H),2.16-2.27(m,1H),2.35-2.41(m,2H),2.76-2.83(m,2H),2.97(d,J=4.4Hz,3H),4 .68(bs,1H),6.29(dd,J=14.3,1.8Hz,1H),6.48(dd,J=8.3,1.8Hz,1H),6.75(q,J=4.4Hz,1H),7.90(dd,J=8.3,8.3Hz,1H); 13C NMR (CDCl3, 100MHz) δ15.7, 26.7, 33.9, 49.4, 100.2 (d, J = 29.5Hz), 110.6, 111.0 (d, J = 11. 8Hz), 133.1 (d, J = 4.2Hz), 148.4 (d, J = 12.0Hz), 162.0 (d, J = 244.1Hz), 164.4 (d, J = 3.6Hz).

[0675] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (2.16 g, 9.47 mmol) and N-methyl-4-(1-cyanocyclobutylamino)-2-fluorobenzamide (52c) (1.303 g, 5.27 mmol) in DMF (20 ml) was heated at 80°C under microwave irradiation for 16 hours. Methanol (50 ml) and 2N aqueous HCl (20 ml) were added to this mixture. This second mixture was refluxed for 3 hours. After cooling to room temperature, the reaction mixture was poured into cold water (100 ml) and extracted with ethyl acetate (150 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to give N-methyl-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-2-fluorobenzamide (52d) [RD162] (1.43 g, 3.0 mmol, 57%), the structure of which is shown in Formula 28, as a yellow powder.

[0676]

[0677] 1 H NMR(CDCl3,400MHz)δ1.65-1.75(m,1H),2.18-2.30(m,1H),2.49-2.57(m,2 H),2.67-2.73(m,2H),3.07(d,J=4.4Hz,3H),6.75(q,J=4.6Hz,1H),7.17(dd ,J=11.5,1.9Hz,1H),7.26(dd,J=8.3,1.9Hz,1H),7.83(dd,J=8.2,2.0Hz,1 H),7.95(d,J=1.8Hz,1H),7.97(d,J=8.3Hz,1H)8.30(dd,J=8.3,8.3Hz,1H); 13C NMR(CDCl3,100MHz)δ13.6,27.0,31.7,67.4,110.3,114.8,118.2,118.5,121.9(q,J=272.7Hz),126.6,127.0(q,J=4.8Hz),13 2.1, 133.3 (q, J = 33.2Hz), 133.8, 135.3, 136.8, 139.1 (d, J = 10.9Hz), 160.5 (d, J = 249.1Hz), 162.7 (d, J = 3.3Hz), 174.3, 179.8; 19 F NMR (CDCl3, 100MHz) δ-111.13,-62.58.

[0678] Example 53 [RD163]

[0679] A mixture of 4-nitro-3-fluorophenol (0.314 g, 2 mmol) and iron powder (0.56 g, 10 mmol) in ethyl acetate (4 ml) and acetic acid (2 ml) was refluxed for 3 hours. The solid particles were filtered off. The filtrate was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated to give 4-amino-3-fluorophenol (53a) (0.25 g, 19.6 mmol, 98%) as a brown solid.

[0680] 1 H NMR (CDCl3, 400MHz) δ6.48-6.58 (m, 2H), 6.61-6.70 (m, 1H), 7.87 (bs, 3H).

[0681] Sodium cyanide (0.194 g, 4 mmol) was added to a mixture of 4-amino-3-fluorophenol (0.29 g, 2.28 mmol) and cyclobutanone (0.175 g, 2.5 mmol) in 90% acetic acid (3 ml). The reaction mixture was stirred at room temperature for 15 hours. The mixture was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 90:10) to afford 1-(2-fluoro-4-hydroxyphenylamino)-cyclobutanecarbonitrile (53b) (0.271 g, 1.31 mmol, 58%) as an off-white solid.

[0682] 1H NMR(CDCl3,400MHz)δ2.13-2.20(m,2H),2.36-2.41(m,2H),2.70.2.75(m,2H),4.00(bs,1H),6.46(bs,1H) ,6.52(ddd,J1=2.2Hz,J2=0.65Hz,J3=0.22Hz,1H),6.57(d,J=2.3Hz),6.62(dd,J1=3.0Hz,J2=0.67Hz,1H); 13 C NMR (CDCl3, 100MHz) δ15.7, 34.1, 50.9, 104.0 (d, J = 21.9Hz), 111.0 (d, J = 3.4Hz), 115.8 (d, J = 3.7Hz), 121.8, 125.3 (d, J = 12.3Hz), 150.1 (d, J = 10.4Hz), 152.8 (d, J = 239.3Hz).

[0683] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.228 g, 1.0 mmol) and 1-(2-fluoro-4-hydroxyphenylamino)-cyclobutanecarbonitrile (53b) (0.145 g, 0.7 mmol) in dry DMF (2 ml) was stirred at room temperature for 24 hours. Methanol (10 ml) and 2M aqueous HCl (2 ml) were added to this mixture. This second mixture was refluxed for 1 hour. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (50 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (neat dichloromethane, then dichloromethane:acetone, 90:10) to afford 4-[5-(2-fluoro-4-hydroxyphenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl]-2-trifluoromethylbenzonitrile (53c) [RD163] (0.17 g, 0.39 mmol, 56%), the structure of which is shown in Formula 29, as an off-white powder.

[0684]

[0685] 1 H NMR(CDCl3,400MHz)δ1.66-1.75(m,1H),2.18-2.28(m,1H),2.42-2.50(m,1H),2.54-2.67(m,3H),6.76(d,J=2.2Hz,2H) ,7.15(t,J=2.1Hz,1H),7.35(bs,1H),7.87(dd,J1=8.2Hz,J2=1.8Hz,1H),7.97(d,J=8.2Hz,1H),7.98(d,J=1.8Hz,1H); 13C NMR(CDCl3,100MHz)δ13.8,31.0,67.6,104.8(d,J=22.3Hz),109.8,112.6,114.4(d,J=13.1Hz),114.9,121.9(q,J=272.8Hz),1 27.1(q,J=4.8Hz),132.0,132.3,133.5(q,J=33.3Hz),135.3,137.2,159.3(d,J=11.2Hz),159.6(d,J=249.7Hz),175.2,180.5; 19 F NMR (CDCl3, 100MHz) δ-117.5,-62.49.

[0686] Example 54 [RD168]

[0687] A mixture of 4-nitro-2-fluorobenzonitrile (1.83 g, 5 mmol) and iron powder (1.68 g, 6 mmol) in acetic acid (40 ml) and ethyl acetate (40 ml) was refluxed for 2 hours. The solid was filtered off, and the filtrate was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 95:5) to afford 4-amino-2-fluorobenzonitrile (54a) (0.653 g, 4.8 mmol, 96%).

[0688] Sodium cyanide (0.74 g, 15 mmol) was added to a mixture of 4-amino-2-fluorobenzonitrile (1.36 g, 10 mmol) and cyclopentanone (1.26 g, 15 mmol) in 90% acetic acid (10 ml). The reaction mixture was stirred at room temperature for 3 hours, then the mixture was heated to 80°C and stirred for an additional 5 hours. The mixture was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 97:3) to afford 4-(1-cyanocyclopentylamino)-2-fluorobenzonitrile (54b) (2.07 g, 9.03 mmol, 90%) as a yellow solid.

[0689] 1 H NMR (CDCl3, 400MHz) δ1.69-1.91 (m, 4H), 2.13-2.18 (m, 2H), 2.37-2.42 (m, 2H), 5.08 (bs, 1H), 6.54-6.62 (m, 2H), 7.39 (t, J = 7.3Hz, 1H); 13C NMR (CDCl3, 100MHz) δ23.7, 39.8, 56.8, 89.6 (d, J = 15.8Hz), 101.2 (d, J = 23.8Hz), 11 0.9, 115.2, 120.8, 134.1 (d, J = 2.4Hz), 150.3 (d, J = 11.2Hz), 164.5 (d, J = 254.1Hz).

[0690] A mixture of 4-isothiocyanato-2-trifluoromethylbenzonitrile (1a) (0.171 g, 0.75 mmol) and 4-(1-cyanocyclopentylamino)-2-fluorobenzonitrile (54b) (0.115 g, 0.5 mmol) in dry DMF (1 ml) was heated at 60°C under microwave irradiation for 48 hours. Methanol (3 ml) and 2M aqueous HCl (2 ml) were added to this mixture. This second mixture was refluxed for 1 hour. After cooling to room temperature, the reaction mixture was poured into cold water (10 ml) and extracted with ethyl acetate (15 ml). The organic layer was dried over magnesium sulfate, concentrated, and purified by chromatography (dichloromethane:acetone, 98:2) to afford 4-[1-(4-cyano-3-fluorophenyl)-4-oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-3-yl]-2-trifluoromethylbenzonitrile (54c) [RD168] (0.017 g, 0.037 mmol, 7%), the structure of which is shown in Formula 30, as an off-white powder.

[0691]

[0692] 1 H NMR(CDCl3,400MHz)δ1.53-1.63(m,2H),1.89-2.00(m,2H),2.09-2.16(m,2H),2.35-2.42( m,2H),7.27-7.37(m,2H),7.78-7.90(m,3H),7.95(d,J=1.8Hz,1H),7.97(d,J=8.3Hz,1H); 13 C NMR(CDCl3,100MHz)δ25.2,36.5,75.3,103.2(d,J=15.3Hz),110.4,112.8,114.7,119.2(d,J=20.7Hz),121.9(q,J=272.8Hz),127 .0(q,J=4.8Hz),132.1,133.7(q,J=33.2Hz),134.6,135.3,135.8,136.8,141.8(d,J=9.5Hz),163.4(d,J=261.5Hz),175.3,180.1.

[0693] Example 55 [RD136 and RD142]

[0694] Additional diarylhydantoin compounds can be synthesized, including the following compounds shown in Formulas 35 and 36.

[0695]

[0696]

[0697] Example 56 [RD162']

[0698] In the following, reactions that are sensitive to air or moisture were performed under a nitrogen atmosphere using oven-dried glassware and standard syringe / septa techniques. Reactions were monitored using SiO2 TLC plates under UV light (254 nm) followed by visualization with p-anisaldehyde or ninhydrin colorimetric solutions. Column chromatography purification was performed on silica gel 60. Unless otherwise stated, 1 H NMR spectra were measured in CDCl3 at 400 MHz, and the data are given in ppm (δ), which are chemical shifts (multiplicity, integration, and coupling constants are in Hz) relative to an internal standard (TMS, 0.0 ppm).

[0699]

[0700] Periodic acid (1.69 g, 7.41 mmol) was dissolved in acetonitrile (25 mL) with vigorous stirring, and chromium trioxide (0.16 g, 1.60 mmol) was then dissolved in the resulting solution. 2-Fluoro-4-nitrotoluene (0.33 g, 2.13 mmol) was added to the above solution with stirring. A white precipitate immediately formed due to the exothermic reaction. After stirring for 1 hour, the supernatant of the reaction mixture was poured into a flask and the solvent was evaporated. The residue was extracted with dichloromethane (2 × 30 mL) and water (2 × 30 mL). The organic layer was dried over magnesium sulfate and concentrated to yield 2-fluoro-4-nitrobenzoic acid (Formula 37) (0.32 mg, 81%) as a white solid. 1 H NMR δ 8.06 (ddd, 1H, J = 9.9, 2.2 and 0.3), 8.13 (ddd, 1H, J = 8.6, 2.2 and 0.9), 8.25 (ddd, 1H, J = 8.6, 7.0 and 0.3).

[0701]

[0702] Thionyl chloride (0.15 g, 1.30 mmol) was slowly added to a solution of 2-fluoro-4-nitrobenzoic acid (Formula 37) (0.20 g, 1.10 mmol) in DMF (5 mL) cooled to -5°C. The mixture was stirred at -5°C for another hour. Excess methylamine (freshly prepared, distilled from a 40% aqueous solution thereof) was added to the reaction mixture. This second mixture was stirred for another hour. Ethyl acetate (50 mL) was added to the mixture, which was washed with brine (2 x 50 ml). The organic layer was dried over magnesium sulfate and concentrated to give N-methyl-2-fluoro-4-nitrobenzamide (Formula 38) (0.18 g, 85%) as a light yellow solid. 1 H NMR (acetone-d6) δ 3.05 (d, 3H, J = 4.3), 6.31 (dd, 1H, J = 13.5 and 2.1), 6.40 (dd, 1H, J = 8.6 and 2.1), 7.64 (dd, 1H, J = 8.6 and 8.6).

[0703]

[0704] A mixture of N-methyl-2-fluoro-4-nitrobenzamide (Formula 38) (0.18 g, 0.91 mmol) and iron powder (0.31 g, 5.60 mmol) in ethyl acetate (5 mL) and acetic acid (5 mL) was refluxed for 1 hour. Solid particles were filtered off. The filtrate was washed with water and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. The residue was purified by SiO2 column chromatography (dichloromethane:acetone, 95:5) to afford N-methyl-2-fluoro-4-aminobenzamide (Formula 39) (0.14 g, 92%) as an off-white solid. 1 HNMR (acetone-d6) δ 2.86 (d, 3H, J = 4.3), 5.50 (br s, 2H), 6.37 (dd, 1H, J = 14.7 and 2.1), 6.50 (dd, 1H, J = 8.6 and 2.1), 7.06 (br s, 1H), 7.68 (dd, 1H, J = 8.8 and 8.8).

[0705]

[0706] A mixture of N-methyl-2-fluoro-4-aminobenzamide (Formula 39) (96 mg, 0.57 mmol), acetone cyanohydrin (0.3 mL, 3.14 mmol), and magnesium sulfate (50 mg) was heated to 80°C and stirred for 12 hours. Ethyl acetate (25 mL) was added to the mixture, which was then washed with water (2 x 25 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by SiO2 column chromatography (dichloromethane:acetone, 95:5) to provide N-methyl-2-fluoro-4-(1,1-dimethyl-cyanomethyl)-aminobenzamide (Formula 40) (101 mg, 75%) as a white solid. 1 H NMR δ 1.74 (s, 6H), 2.98 (dd, 3H, J = 4.8 and 1.1), 6.58 (dd, 1H, J = 14.6 and 2.3), 6.63 (dd, 1H, J = 8.7 and 2.3), 6.66 (br s, 1H), 7.94 (dd, 1H, J = 8.7 and 8.7).

[0707]

[0708] 4-Amino-2-trifluoromethylbenzonitrile (2.23 g, 12 mmol) was added dropwise to a well-stirred heterogeneous mixture of thiophosgene (1 mL, 13 mmol) in water (22 mL) at room temperature over 15 minutes. Stirring was continued for an additional hour. The reaction mixture was extracted with chloroform (3 x 15 mL). The combined organic phases were dried over magnesium sulfate and evaporated to dryness under reduced pressure to afford the desired product, 4-isothiocyanato-2-trifluoromethylbenzonitrile (Formula 41), as a brown solid, which was used directly in the next step (2.72 g, 11.9 mmol, 99%). 1 H NMR δ 7.49 (dd, 1H, J = 8.3 and 2.1), 7.59 (d, 1H, J = 2.1), 7.84 (d, 1H, J = 8.3).

[0709]

[0710] 56-1)RD162'

[0711] A mixture of N-methyl-2-fluoro-4-(1,1-dimethyl-cyanomethyl)-aminobenzamide (Formula 40) (30 mg, 0.13 mmol) and 4-isothiocyanato-2-trifluoromethylbenzonitrile (Formula 41) (58 mg, 0.26 mmol) in DMF (1 mL) was heated at 100°C for 11 hours under microwave irradiation. Methanol (20 mL) and 1N aqueous HCl (5 mL) were added to this mixture. This second mixture was refluxed for 1.5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by SiO2 column chromatography (dichloromethane:acetone, 95:5) to obtain RD162' (Formula 42) (15 mg, 25%) as colorless crystals. 1 H NMR δ 1.61 (s, 6H), 3.07 (d, 3H, J = 4.1), 6.71 (m, 1H), 7.15 (dd, 1H, J = 11.7 and 2.0), 7.24 (dd, 1H, J = 8.4 and 2.0), 7.83 (dd, 1H, J = 8.2 and 2.1), 7.95 (d, 1H, J = 2.1), 7.99 (d, 1H, J = 8.2), 8.28 (dd, 1H, J = 8.4 and 8.4).

[0712] Example 57

[0713]

[0714] A mixture of N-methyl-2-fluoro-4-aminobenzamide (Formula 39) (62 mg, 0.37 mmol), cyclopentanone (0.07 mL, 0.74 mmol), and TMSCN (0.1 mL, 0.74 mmol) was heated to 80°C and stirred for 13 hours. Ethyl acetate (2 x 20 mL) was added to the mixture and then washed with water (2 x 20 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 95:5) to provide N-methyl-2-fluoro-4-(1-cyanocyclopentyl)aminobenzamide (Formula 43) (61 mg, 63%) as a white solid. 1 H NMRδ7.95(dd,1H,J=8.8,8.8Hz),6.65(br s,1H),6.59(dd,1H,J=8.8,2.3Hz),6.50(dd,1H,J=14.6,2.3Hz),4.60(br s,1H),2.99(dd,3H,J=4.8,1.1Hz),2.36-2.45(m,2H),2.10-2.18(m,2H),1.82-1.95(m,4H).

[0715]

[0716] 57-1)RD162"

[0717] A mixture of N-methyl 2-fluoro-4-(1-cyanocyclopentyl)aminobenzamide (Formula 43) (57 mg, 0.22 mmol) and 4-isothiocyanato-2-trifluoromethylbenzonitrile (0.15 g, 0.65 mmol) in DMF (3 mL) was heated at 130°C for 12 hours under microwave irradiation (open vessel). Methanol (20 mL) and 1N aqueous HCl (5 mL) were added to this mixture. This second mixture was refluxed for 1.5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 95:5) to give 4-(3-(4-cyano-3-(trifluoromethyl)phenyl)-4-oxo-2-thioxo-1,3-diazaspiro[4.4]nonan-1-yl)-2-fluoro-N-methylbenzamide, RD162" (Formula 44) (8 mg, 7%) as a light yellow solid. 1 H NMRδ8.28(dd,1H,J=8.4,8.4Hz),7.98(d,1H,J=8.3Hz),7.96(d,1H,J=1.8Hz),7.84(dd,1H,J=8.3,1.8Hz),7.27(dd,1H,J=8.4,1.8Hz),7.17( dd,1H,J=11.7,1.8Hz),6.67-6.77(m,1H),3.07(d,3H,J=4.3Hz),2.32 -2.41(m,2H),2.13-2.21(m,2H),1.85-1.96(m,2H),1.49-1.59(m,2H).

[0718] Example 58

[0719]

[0720] Trifluoroacetic anhydride (0.85 mL, 6.14 mmol) was added to a solution of 4-(4-aminophenyl)butyric acid (0.5 g, 2.79 mmol) in chloroform (10 mL) at 0°C. The mixture was warmed to room temperature and stirred for 3 hours. The mixture was partitioned between chloroform (20 mL) and water (20 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 9:1) to yield 4-[4-(2,2,2-trifluoroacetylamino)phenyl]butyric acid (Formula 45) (0.53 g, 69%). 1HNMR δ7.81 (br s, 1H), 7.48 (d, 2H, J = 8.5Hz), 7.22 (d, 2H, J = 8.5Hz), 2.68 (t, 2H, J = 7.5Hz), 2.38 (t, 2H, J = 7.5Hz), 1.96 (p, 2H, J = 7.5Hz).

[0721]

[0722] Thionyl chloride (71 mg, 0.60 mmol) was slowly added to a solution of 4-[4-(2,2,2-trifluoroacetylamino)phenyl]butyric acid (Formula 45) (0.15 g, 0.55 mmol) in DMF (5 mL) cooled to -5°C. The mixture was stirred at -5°C for another hour. Excess dimethylamine (freshly prepared, distilled from a 40% aqueous solution) was added to the reaction mixture. This second mixture was stirred for another hour. Ethyl acetate (50 mL) was added to the mixture, which was washed with brine (2 x 50 ml). The organic layer was dried over magnesium sulfate and concentrated to provide N,N-dimethyl 4-[4-(2,2,2-trifluoroacetylamino)phenyl]butyramide (Formula 46) (0.17 g, quantitative) as a light yellow solid. 1 H NMRδ9.70(br s,1H),7.55(d,2H,J=8.6Hz),7.11(d,2H,J=8.6Hz),2.91(s,3H),2.89(s ,3H), 2.60(t,2H,J=7.7Hz), 2.27(t,2H,J=7.7Hz), 1.89(p,2H,J=7.7Hz).

[0723]

[0724] 1N NaOH solution (3 mL) was added to a solution of N,N-dimethyl-4-[4-(2,2,2-trifluoroacetylamino)phenyl]butanamide (Formula 46) (0.17 g, 0.55 mmol) in methanol (2 mL) at room temperature. The mixture was stirred for 14 hours. The mixture was partitioned between chloroform (25 mL) and water (25 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 9:1) to provide N,N-dimethyl-4-(4-aminophenyl)butanamide (Formula 47) (74 mg, 66%) as a white solid. 1HNMRδ6.97(d,2H,J=8.3Hz),6.61(d,2H,J=8.3Hz),3.56(br s, 2H), 2.92 (s, 6H), 2.56 (t, 2H, J = 7.7Hz), 2.28 (t, 2H, J = 7.7Hz), 1.91 (p, 2H, J = 7.7Hz).

[0725]

[0726] A mixture of N,N-dimethyl-4-(4-aminophenyl)butanamide (Formula 47) (74 mg, 0.36 mmol), cyclobutanone (54 mg, 0.78 mmol), and TMSCN (77 mg, 0.78 mmol) was heated to 80°C and stirred for 15 hours. Ethyl acetate (2 x 20 mL) was added to the mixture and then washed with water (2 x 20 ml). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 9:1) to provide N,N-dimethyl-4-[4-(1-cyanocyclobutylamino)phenyl]butanamide (Formula 48) (58 mg, 57%) as a white solid. 1 H NMRδ7.07(d,2H,J=8.5Hz),6.59(d,2H,J=8.5Hz),3.94(br s,1H),2.94(s,3H),2.93(s,3H),2.75-2.83(m,2H),2.60(t,2H,J=7.6Hz),2.3 3-2.42(m,2H),2.30(t,2H,J=7.6Hz),2.11-2.28(m,2H),1.93(p,2H,J=7.6Hz).

[0727]

[0728] A mixture of N,N-dimethyl-4-[4-(1-cyanocyclobutylamino)phenyl]butanamide (Formula 48) (58 mg, 0.20 mmol) and 4-isothiocyanato-2-trifluoromethylbenzonitrile (74 mg, 0.32 mmol) in DMF (3 mL) was heated at reflux for 2 hours. Methanol (20 mL) and 1N aqueous HCl (5 mL) were added to this mixture. This second mixture was refluxed for 1.5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 95:5) to give 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)-N,N-dimethylbutanamide, RD169 (Formula 49) (44 mg, 42%) as a light yellow solid. 1 H NMRδ7.98(s,1H),7.97(d,1H,J=8.2Hz),7.86(d,1H,J=8.2Hz),7.42(d,2H,J=8.3Hz),7.22(d,2H,J=8.3Hz),2.99(s,3H),2.96(s,3H),2.78 (t,2H,J=7.5Hz),2.62-2.70(m,2H),2.52-2.63(m,2H),2.40(t,2H,J=7.5Hz),2.15-2.30(m,1H),2.04(p,2H,J=7.5Hz),1.62-1.73(m,1H).

[0729] Example 59

[0730]

[0731] A mixture of 4-(4-aminophenyl)butyric acid (0.20 g, 1.12 mmol), cyclobutanone (0.17 mL, 2.23 mmol), and TMSCN (0.30 mL, 2.23 mmol) was heated to 80°C and stirred for 13 hours. Ethyl acetate (2 x 30 mL) was added to the mixture, followed by washing with water (2 x 30 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 9:1) to yield 4-[4-(1-cyanocyclobutylamino)phenyl]butyric acid (Formula 50) (0.21 g, 74%) as a pale yellow solid. 1H NMRδ7.06(d,2H,J=8.6Hz),6.59(d,2H,J=8.6Hz),2.75-2.83(m,2H),2.59(t,2H,J=7.5 Hz), 2.37 (t, 2H, J = 7.5Hz), 2.33-2.42 (m, 2H), 2.11-2.28 (m, 2H), 1.92 (p, 2H, J = 7.5Hz).

[0732]

[0733] A mixture of 4-[4-(1-cyanocyclobutylamino)phenyl]butanoic acid (Formula 50) (0.21 g, 0.83 mmol) and 4-isothiocyanato-2-trifluoromethylbenzonitrile (0.25 g, 1.08 mmol) in toluene (10 mL) was heated at reflux for 1 hour. 1N aqueous HCl (5 mL) was added to this mixture. This second mixture was refluxed for 1.5 hours. After cooling to room temperature, the reaction mixture was poured into cold water (50 mL) and extracted with ethyl acetate (50 mL). The organic layer was dried over magnesium sulfate and concentrated, and the residue was purified by silica gel column chromatography (dichloromethane:acetone, 95:5) to yield 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanoic acid, RD141 (Formula 51) (60 mg, 15%). 1 H NMRδ7.98(d,1H,J=1.8Hz),7.97(d,1H,J=8.3Hz),7.86(dd,1H,J=8.3,1.8Hz),7.42(d,2H,J=8.5Hz),7.24(d,2H,J=8.5Hz),2.79(t,2 H,J=7.5Hz),2.62-2.68(m,2H),2.51-2.59(m,2H),2.47(t,2H,J=7.5Hz),2.14-2.26(m,1H),2.06(p,2H,J=7.5Hz),1.60-1.70(m,1H).

[0734] Example 60

[0735]

[0736] To a solution of 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanoic acid, RD141 (Formula 51) (60 mg, 0.12 mmol) in DMF (3 mL) at 0°C was added thionyl chloride (0.01 mL, 0.15 mmol). The mixture was stirred at 0°C for 1 hour. Ammonia was then bubbled into the mixture. The mixture was partitioned with ethyl acetate (25 mL) and water (25 mL). The organic layer was dried over magnesium sulfate, concentrated and purified by chromatography (dichloromethane:acetone, 70:30) to give 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanamide, RD130 (Formula 52) (37 mg, 61%) as a white powder. 1 H NMRδ7.97(d,1H,J=1.8Hz),7.95(d,1H,J=8.3Hz),7.85(dd,1H,J=8.3Hz),7.39(d,2H,J=8.3Hz),7.22(d,2H,J=8.3Hz),5.59(br s,2H),2.77(t,2H,J=7.5Hz),2.62-2.68(m,2H),2.51-2.59(m,2H),2.31(t ,2H,J=7.5Hz),2.16-2.25(m,1H),2.05(p,2H,J=7.5Hz),1.57-1.70(m,1H).

[0737] Example 61

[0738]

[0739] A solution of DMSO (0.01 mL, 0.12 mmol) in dry dichloromethane (1 mL) was added to a stirred solution of oxalyl chloride (0.01 mL, 0.09 mmol) in dry dichloromethane (2 mL) at -78°C. After 15 minutes, a solution of 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanamide, RD130 (Formula 52) (35 mg, 0.07 mmol) in dichloromethane was added to the reaction mixture. Stirring was continued at -78°C for 20 minutes, followed by the addition of triethylamine (0.03 mL, 0.22 mmol). After 30 minutes at -78°C, the reaction mixture was warmed to room temperature and quenched with saturated aqueous NH4Cl. The reaction mixture was diluted with dichloromethane and extracted with dichloromethane. The organic layer was dried over magnesium sulfate, concentrated and purified by chromatography (dichloromethane:acetone, 95:5) to afford 4-(5-(4-(3-cyanopropyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-7-yl)-2-(trifluoromethyl)benzonitrile, RD170 (Formula 53) (29 mg, 87%) as a viscous oil. 1 H NMRδ7.98(d,1H,J=1.8Hz),7.98(d,1H,J=8.3Hz),7.86(dd,1H,J=8.3,1.8Hz),7.43(d,2H,J=8.4Hz),7.27(d,2H,J=8.4Hz),2.90(t,2 H,J=7.3Hz),2.63-2.73(m,2H),2.52-2.62(m,2H),2.42(t,2H,J=7.3Hz),2.18-2.30(m,1H),2.07(p,2H,J=7.3Hz),1.63-1.73(m,1H).

[0740] Those skilled in the art can modify and / or combine the syntheses described herein to prepare other diarylhydantoin compounds.

[0741] Inventive compounds also include those having the formula:

[0742]

[0743] wherein R is selected from hydrogen, aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, halogen, SO2R 11 NR 11R 12 NR 12 (CO)OR 11 、NH(CO)NR 11 R 12 NR 12 (CO)R 11 、O(CO)R 11 、O(CO)OR 11 、O(CS)R 11 NR 12 (CS)R 11 、NH(CS)NR 11 R 12 NR 12 (CS)OR 11 .

[0744] R1 and R2 are independently selected from hydrogen, aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl.

[0745] R1 and R2 may be linked to form a ring, and the ring may be a heterocyclic group, a substituted heterocyclic group, a cycloalkyl group, or a substituted cycloalkyl group.

[0746] R3 is selected from aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, SO2R 11 NR 11 R 12 、(CO)OR 11 、(CO)NR 11 R 12 、(CO)R 11 、(CS)R 11 、(CS)R 11 、(CS)NR 11 R 12 、(CS)OR 11 .

[0747] R5 is CN or NO2 or SO2R 11 .

[0748] R6 is CF3, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, or halogen.

[0749] A is a sulfur atom (S) or an oxygen atom (O).

[0750] B is O or S or NR3.

[0751] X is carbon or nitrogen and may be located at any position on the ring.

[0752] R 11 and R 12 independently selected from hydrogen, aryl, aralkyl, substituted aralkyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl.

[0753] R 11 and R 12 They may be linked to form a ring which may be an aromatic heterocyclic group or a non-aromatic heterocyclic group, a substituted aromatic heterocyclic group, a cycloalkyl group, or a substituted cycloalkyl group.

[0754] Pharmacological testing of compounds

[0755] The compounds described in the above synthetic routes were screened for AR antagonistic and agonistic activity on hormone-refractory prostate cancer cells using screening methods similar to those described in PCT applications US04 / 42221 and US05 / 05529 (incorporated herein by reference). Many compounds showed potent antagonistic activity against AR overexpressed in hormone-refractory prostate cancer with minimal agonistic activity.

[0756] In vitro bioassay

[0757] Testing the effects of compounds on AR via receptors

[0758] The compounds were tested using an artificial AR response reporter system in a hormone-refractory prostate cancer cell line. In this system, prostate cancer LNCaP cells are genetically modified to stably express AR levels approximately 5 times higher than endogenous levels. Exogenous AR has properties similar to endogenous AR, and both are stabilized by the synthetic androgen R1881. AR-overexpressing cells are also genetically modified to stably incorporate AR response reporters, and the reporter activity of these cells shows the characteristics of hormone-refractory prostate cancer. It corresponds to low concentrations of the synthetic androgen R1881, only suppressed by high concentrations of bicalutamide (see Table 1), and shows the agonist activity using bicalutamide ( Figure 1 and Table 2). Consistent with published data, bicalutamide inhibited AR-responsive reporters and had no agonist activity in hormone-sensitive prostate cancer cells ( Figure 2 ).

[0759] We tested the antagonist activity of the synthetic compounds described above in the presence of 100 pM R1881. Genetically modified LNCaP cells (LNCaP-AR, also abbreviated as LN-AR) were maintained in Iscove's medium containing 10% fetal bovine serum (FBS). Two days prior to drug treatment, cells were grown in Iscove's medium containing 10% charcoal-desorbed FBS (CS-FBS) to remove androgens. Cells were split and grown in Iscove's medium containing 10% CS-FBS, 100 pM R1881, and increasing concentrations of the test compound. After two days of incubation, reporter activity was assayed.

[0760] Table 1 lists the IC50 values ​​of these compounds for inhibiting AR in hormone-refractory prostate cancer. The IC50 of the control substance bicalutamide is 889 nM. In inhibiting AR in hormone-refractory prostate cancer, most of the identified compounds (diarylthiohydantoins) have IC50 values ​​between 100-200 nM. In contrast, the antiandrogenic compounds listed in the examples of U.S. Patent No. 5,705,654, such as Examples 30-2, 30-3, 31-2, 31-3 and 24-3 (RD73-RD77), have no inhibitory activity against AR in this system.

[0761] Table 1

[0762] Antagonistic activity against AR in hormone-refractory prostate cancer, measured by AR-responsive reporter and endogenous PSA expression.

[0763]

[0764]

[0765]

[0766]

[0767]

[0768] (*) No: The compound does not inhibit the AR response reporter; (**) n / a: The compound was not tested in this assay.

[0769] A previously unrecognized property of AR overexpression in hormone-refractory prostate cancer is its ability to convert antagonism to agonism. Therefore, only compounds with minimal or no agonist activity are suitable as anti-androgens for this disease. In order to determine the agonist activity of different compounds, we used AR-responsive reporters as a metric in the LN-AR system to detect their stimulatory activity on AR in the absence of R1881. Table 2 lists the agonist activity of different compounds. Consistent with previous results, bicalutamide activated AR in hormone-refractory prostate cancer. Diarylthiohydantoin derivatives such as Examples 7-3b (RD37), 33 (RD91), 34 (RD92) and 35 (RD93) had no agonist activity. In contrast, RU59063 and other antiandrogen compounds, such as those exemplified in US Pat. No. 5,705,654, eg, Examples 30-2, 30-3, 31-2, 31-3, and 24-3 (RD73-RD77), strongly activate AR in hormone-refractory prostate cancer.

[0770] Table 2

[0771] Agonist activity of selected test compounds on AR-responsive reporters in hormone-refractory prostate cancer by fold induction at increasing concentrations of the compounds

[0772]

[0773]

[0774] (*) Fold induction: activity induced by a specific test compound relative to the activity in DMSO vehicle; (**) n / a: compound was not tested in this assay.

[0775] To test the specificity of AR inhibitors, selected compounds were tested in LNCaP cells with overexpression of the glucocorticoid receptor (GR), the closest member of the AR in the nuclear receptor family. These cells also carry GR response reporters, and the reporter activity is induced by dexamethasone (GR agonist), and GR excitability and induction are blocked by RU486 (GR inhibitor). Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) has no effect on the GR in this system.

[0776] Effects of compounds on AR, as measured by prostate-specific antigen (PSA) secretion levels

[0777] It has been established that PSA levels are indicators of AR activity in prostate cancer. In order to detect whether these compounds affect AR function in a physiological environment, in AR-overexpressed LNCaP cells (LNCaP-AR, also abbreviated as LN-AR), we determined the secretion level of endogenous PSA induced by R1881. LNCaP-AR cells are lymph node tumor lineages of prostate cells transduced with a plasmid expressing androgen receptor. LNCaP-AR cells are maintained in Iscove's medium containing 10% FBS. 2 days before drug treatment, cells are grown in Iscove's medium containing 10% CS-FBS to remove androgens. The cells are differentiated and grown in Iscove's medium containing 10% CS-FBS and test compounds with an appropriate concentration of R1881. After 4 days of cultivation, secreted PSA levels are tested using a PSA ELISA kit (American Qualex, San Clemente, CA).

[0778] Secreted PSA levels in LNCaP-AR cells were strongly induced by 25 pM of R1881. In contrast, PSA in maternal LNCaP cells was not induced until the concentration of R1881 reached 100 pM. This is consistent with our previous report that AR in hormone-refractory prostate cancer is hypersensitive to androgens. Dose-dependent inhibition of AR activity was performed to determine the IC50 values ​​of various compounds for inhibiting PSA expression, and the results are listed in Table 1. The IC50 values ​​of selected compounds for PSA expression were very similar to those determined using the reporter assay, confirming that diarylhydantoin derivatives are potent inhibitors of AR in hormone-refractory prostate cancer.

[0779] Using secreted PSA as a surrogate marker, we also tested the agonist activity of selected compounds on the AR in hormone-refractory prostate cancer. To this end, androgen-deficient (ndrogen-starved) AR-overexpressing LNCaP cells were cultured in the absence of R1881 with increasing concentrations of the synthetic compounds described above, and secreted PSA in the culture medium was measured after 4 days.

[0780] Table 3 lists the agonist activity of selected compounds. Consistent with the results obtained in the reporter assay, diarylthiohydantoin derivatives such as Examples 7-3b (RD37), 33 (RD91), 34 (RD92), and 35 (RD93) had no agonist activity. In contrast, RU59063 and other antiandrogenic compounds such as those listed in the examples of U.S. Patent No. 5,705,654, such as Examples 30-2 (RD73), 30-3 (RD74), and 31-2 (RD75), stimulated PSA expression in hormone-refractory prostate cancer.

[0781] Table 3

[0782] Agonist activity of selected test compounds on endogenous PSA in hormone-refractory prostate cancer by fold induction at increasing concentrations of the compounds

[0783]

[0784]

[0785] (*) Fold induction: activity induced by a specific test compound relative to the activity in DMSO vehicle; (**) n / a: compound was not tested in this assay.

[0786] Effects of compounds on AR mitochondrial activity, as assessed by MTS assay

[0787] LNCaP-AR cells were maintained in Iscove's medium containing 10% FBS. Compounds were tested for their effects on the growth of hormone-refractory prostate cancer cells. Overexpressing LNCaP cells were used because these cells are equivalent to hormone-refractory prostate cancer cells in vitro and in vivo (1). We measured mitochondrial activity, a surrogate for growth, by the MTS assay. LNCaP cells with overexpressed AR (LN-AR) were maintained in Iscove's medium containing 10% FBS. Two days before drug treatment, cells were grown in Iscove's medium containing 10% CS-FBS to remove androgens. The cells were then differentiated and grown in Iscove's medium containing 10% CS-FBS with an appropriate concentration of R1881 and increasing concentrations of the test compound. After 4 days of culture, cell growth was monitored using MTS (Promega, Madison, WI).

[0788] Consistent with the reporter assay and PSA assay, the growth of AR-overexpressing LNCaP was stimulated by 25 mM R1881, but the parental cells were not stimulated until the R1881 concentration reached 100 mM. Figure 2 The inhibitory effect of selected compounds on the growth of hormone-refractory prostate cancer in the presence of 100 pM of R1881 is shown. The current clinical drug bicalutamide does not inhibit hormone-refractory prostate cancer. In contrast, Example 5-3b (RD7) (4-[3-(4-methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethyl-benzonitrile) and Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) inhibit hormone-refractory prostate cancer with high efficacy.

[0789] We tested whether growth inhibition could occur by targeting AR in the MTS assay. Example 5-3b (RD7) (4-[3-(4-methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethyl-benzonitrile) and Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) were tested in DU-145 cells, a prostate cancer cell line that lacks AR expression. These compounds had no growth inhibitory effect on DU-145 cells. In addition to AR-expressing prostate cancer cells, the compounds did not inhibit the growth of other cells and had no effect on MCF7 and SkBr3 (two frequently used breast cancer cells) or 3T3 (a common mouse fibroblast cell line).

[0790] Examples of the in vitro biological activities of diarylthiohydantoin derivatives are shown in Figure 3 、 4 and 5. For example, based on relative luciferase activity, Figure 3 The compounds are shown to be ranked in order of greatest to least activity at a concentration of 500 nM as follows: RD152 > RD153 > RD145 > RD163 > RD161 = RD162 > bicalutamide. For example, based on relative PSA levels, Figure 4 The order of most to least active compounds at a concentration of 500 nM is shown as follows: RD138 > RD131 > RD37 > RD133 > RD134 > RD137 > RD138 > RD135 > bicalutamide. For example, based on relative MTS units, Figure 5 It is shown that at a concentration of 500 nM, the compounds are ranked in order of most to least active as follows: RD168>RD37>RD141>RD162>bicalutamide.

[0791] Inhibitory effect on hormone-refractory prostate cancer xenografts

[0792] Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) was used to test whether diarylhydantoin derivatives have an effect on hormone-refractory prostate cancer in vivo. We first tested this compound on xenograft tumors established from AR-overexpressing LNCaP cells. The genetically modified cells in Matrigel (Collaborative Biomedical) were injected subcutaneously into the flank of castrated male SCID mice. The three-dimensional size of the tumor was measured weekly with a caliper. After the xenograft tumor was established (the tumor size reached at least 40 mm), the tumor was resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. The tumor was then resuspended in 4% paracetamol. 3) , mice with tumors were randomized and treated with different doses of the compound orally once daily. Consistent with clinical observations, the current clinical drug bicalutamide does not inhibit the growth of hormone-refractory prostate cancer (same as vehicle) (Figure 7a). In contrast, Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) strongly inhibited the growth of these tumors (Figure 7a) and the inhibition was dose-dependent (Figure 7b). In addition, Example 7-3b (RD37) inhibited PSA expression ( Figure 8 ), which is a clinical marker of hormone-refractory prostate cancer.

[0793] Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) was also tested in another hormone-refractory prostate cancer xenograft model, hormone-resistant LAPC4. This model was established from the passage of hormone-sensitive prostate cancer in castrated mice, which mimics the clinical development of prostate cancer (2). Similar to what was found using the AR-overexpressing LNCaP xenograft model, the current clinical drug, bicarb, did not inhibit growth and PSA expression (same as vehicle) in the hormone-resistant LAPC4 xenograft model (Figures 9a and 9b). In contrast, Example 7-3b (RD37) strongly inhibited the growth and PSA expression of these tumors (Figures 9a and 9b).

[0794] Inhibitory effects on hormone-sensitive prostate cancer cells

[0795] To determine whether diarylthiohydantoin derivatives also inhibit hormone-sensitive prostate cancer cells, we tested the effects of selected compounds on LNCaP cell growth by measuring mitochondrial activity (MTS). Unlike the current clinical drug bicalutamide, which has no effect on the growth of hormone-refractory prostate cancer, the current clinical drug bicalutamide moderately inhibits hormone-sensitive LNCaP cells in a dose-dependent manner. Example 5-3b (RD7) (4-[3-(4-methylphenyl)-4,4-dimethyl-5-oxo-2-thioxoimidazolidin-1-yl]-2-trifluoromethyl-benzonitrile) and Example 7-3b (RD37) (4-(8-oxo-6-thioxo-5-(4-methylphenyl)-5,7-diazaspiro[3.4]octan-7-yl)-2-trifluoromethylbenzonitrile) are 10-fold more potent than bicalutamide in inhibiting hormone-sensitive prostate cancer. Figure 10 ).

[0796] In vivo biological testing

[0797] All animal experiments were conducted under the guidance of the Animal Research Committee of the University of California at Los Angeles. Animals were obtained from Taconic and maintained in a laminar flow tower in a defined colony. LNCaP-AR and LNCaP-vector cells were maintained in RPMI medium supplemented with 10% FBS. 10 cells were placed in 100 μl of 1:1 Matrigel / RPMI medium. 6 The cells were injected subcutaneously into the flank of intact or castrated SCID mice. The three-dimensional size of the tumor (length x width x height) was measured using a scale. When the tumor reached approximately 100 mm in size, 3 The mice were randomly divided into groups for treatment. The drug was administered orally at 10 mg / kg and 50 mg / kg per day. To obtain pharmacokinetic readings, animals were photographed using an optical CCD camera 3 hours after the last dose. For the measurement of luciferase activity in photons / second, the ROI was separated from the tumor. The right column shows a representative ROI measurement. Data are shown in Figure 11 and 12. At day 18, RD162 effectively prevented tumor growth and even caused tumor shrinkage, which was significantly more effective than bicalutamide.

[0798] The pharmacokinetics of bicalutamide, 4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-toluene [RD37], N-methyl-4-{4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]phenyl}butanamide [RD131], and N-methyl-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-2-fluorobenzamide (52d) [RD162] were evaluated in vivo using 8-week-old FVB mice purchased from Charles River Laboratories. Mice were divided into three groups at each time point. Two mice were not treated with the drug, and the other two were treated with vehicle solution. Each group was treated with 10 mg per kg body weight.

[0799] The drug is dissolved in a mixture of 1:5:14 of DMSO:PEG400:H20 (carrier solution) and administered to mice via the tail vein. Before processing, animals were warmed under a heating lamp for approximately 20 minutes to dilate their tail veins. Each mouse was placed in a mouse restrainer (Fisher Sci.Cat#01-288-32A) and injected into the dilated tail vein with a carrier solution containing 200 μl of the drug. After drug administration, animals were euthanized at different time points by CO2 inhalation: 5 minutes, 30 minutes, 2 hours, 6 hours, 16 hours. After exposure to CO2, animals were bled immediately by cardiac puncture (BD syringe+27G 5 / 8 needle of 1 ml). For oral doses, the drug was dissolved in a mixture of 50:10:1:989 of DMSO:carboxymethylcellulose:Tween80:H20 by a dosing syringe before oral administration.

[0800] Serum samples were analyzed for drug concentration using HPLC (Waters 600 pump, Waters 600 controller and Waters 2487 detector) equipped with an Alltima C18 column (3μ, 150 mm×4.6 mm). RD37, RD131 and RD162 compounds were detected at a wavelength of 254 nm, and bicalutamide was detected at a wavelength of 270 nm.

[0801] Samples for HPLC analysis were prepared according to the following steps:

[0802] -Separation of blood cells from serum by centrifugation.

[0803] - Add 80 μl of 10 μM internal standard solution and 520 μl of acetonitrile to 400 μl of serum. Precipitation occurs.

[0804] - The mixture was vortexed for 3 minutes and then placed under ultrasound for 30 minutes.

[0805] - Filter out the solid particles or separate them by centrifugation.

[0806] - The filtrate was dried to dryness under a stream of argon. The sample was reconstituted to 80 μl with acetonitrile and then analyzed by HPLC to determine the drug concentration.

[0807] -Use a standard curve for the drug to improve accuracy.

[0808] Figure 13 The concentration of RD162 in serum obtained by intravenous and oral administration is shown in Table 4 as a function of time. Table 4 shows the steady-state concentration (Css) of bicalutamide, RD131 and RD162. The steady-state concentration of RD162 is basically as good as that of bicalutamide and better than RD131.

[0809] Table 4. Steady-state concentrations of bicalutamide, RD131, and RD162 in mouse plasma

[0810]

[0811] Arrangement of compounds within tiers

[0812] Tables 5-10 show that diaryl hydantoin compounds are classified into categories 1-6. Table 11 shows diaryl hydantoin compounds that are not classified. The arrangement of compounds in each category is based on available data and analytical judgment. The data considered include in vitro tests (AR response reporter system in LNCaP cell line, PSA level measurement, MTS mitochondrial test) and in vivo tests (direct measurement or measurement of tumor size by emission induced by luciferase reporter gene, pharmacokinetic test based on plasma levels). Not every compound is tested for every test. All data obtained are not shown. The compounds are arranged relative to each other in the treatment of prostate cancer, especially when two compounds are arranged, they are not subjected to the same experiment. The characteristics considered in establishing the arrangement include AR antagonistic activity, no AR agonism in hormone-resistant cells, prevention of tumor growth, tumor shrinkage, and pharmacokinetic behavior, with a beneficial longer residence time in the blood.

[0813] Category 1

[0814] Generally, Class 1 compounds are diarylthiohydantoins with a disubstituted left aromatic ring, a disubstituted right hydantoin carbon, and an oxygen or nitrogen substituent on the left hydantoin carbon. It is expected that the amide substituent will hydrolyze to oxygen in aqueous solution, such as in biological systems, both in vivo and in vitro. RD100 has good activity because it has an iodine substituent on the left aromatic ring, rather than a CF3 substituent.

[0815] Class 1 compounds (see Table 5) performed much better than bicalutamide in the treatment of prostate cancer. However, RD37 and RD131 were found to be rapidly metabolized, meaning they had a short residence time in the blood. RD162 has desirable pharmacokinetic properties.

[0816] Figure 17 As shown, under the treatment of bicalutamide, the PSA levels of LNCaP cells remained the same or increased relative to those treated with vehicle solution (where the PSA levels were reduced by treatment with RD162). Figure 18 Figure 19 illustrates that under the treatment with carrier solution, tumor size continues to increase.In contrast, under the dosage treatment with 1mg every kg body weight every day with RD162, the growth rate of tumor reduces, and the size of this tumor remains unchanged after about 17 days.Under the dosage treatment with 10mg every kg body weight every day with RD162, tumor size reduces over time.Figure 19 illustrates that under the dosage treatment with 10mg every kg body weight every day with RD162, the photon emission relevant to luciferase activity reduces.Figure 20 has shown that the treatment with this dosage with RD162 has caused the minimizing of tumor size or stabilizing and the reduction of the photon emission relevant to luciferase.

[0817] Figure 21 shows that PSA levels in LN-AR cells were reduced when treated with RD162, RD162', RD162", RD169 and RD170 at doses of 100, 200, 500 and 1000 nM. In addition, the higher the dose, the lower the PSA level. Figure 23 shows the urethra weight and the ratio of photon emission associated with luciferase activity for non-castrated and castrated mice before and after 14 days of treatment with bicalutamide or RD162. Both weight and the ratio of photon emission increased for both non-castrated and castrated mice. Castrated mice treated with RD162 had a decrease in weight and photon emission relative to untreated castrated mice, as did those treated with bicalutamide.

[0818] Therefore, Class 1 compounds are particularly advantageous in being used as AR antagonists and as therapeutic agents in hormone-refractory prostate cancer. They can be used to treat other AR-related diseases or conditions such as benign prostatic hyperplasia, alopecia or acne. These compounds and related compounds can also be used as modulators of other nuclear receptors such as glucocorticoid receptors, estrogen receptors and peroxisome proliferator-activated receptors, and as therapeutic agents for diseases such as breast cancer, ovarian cancer, diabetes, heart disease and metabolic-related diseases that act as nuclear receptors. They are useful, for example, as standards or intermediates or prodrugs in tests.

[0819] Table 5

[0820]

[0821]

[0822]

[0823] Category 2

[0824] Although RD54 was shown to act as an agonist, two classes of compounds (see Table 6) were significantly better than bicalutamide in treating prostate cancer. Figure 3 It was shown that compounds RD145, RD152, RD153, RD162 and RD163 in class 1 and RD161 in class 2 acted to reduce luciferase activity in LNCaP-AR cells at doses ranging from 125 nM to 1000 nM, while control solutions of DMSO and bicalutamide had little or no effect. Figure 4 It was shown that, for example, at a concentration of 1000 nM, compounds RD37 and RD131 from class 1 caused a greater reduction in PSA levels in LNCaP-AR cells than compounds RD133, RD134, and RD138 from class 2. Figure 11 The relationship between tumor volume and time is shown, and it is shown that under treatment with bicalutamide or vehicle solution, the tumor continued to grow, while under treatment with RD162 in Class 1, the tumor size decreased. Figure 12 shows that under treatment with bicalutamide, the photon emission associated with luciferase activity remained approximately the same or increased relative to treatment with vehicle solution, while photon emission decreased under treatment with RD162. Figure 14 It shows that PSA levels were little or not reduced under treatment with bicalutamide, whereas PSA levels were reduced under treatment with RD131 and RD162. Figure 15 shows the IC values ​​of RD37, RD131 and RD162 in Class 1. 50 IC of bicalutamide 50 Much lower.

[0825] Generally, Class 2 compounds are similar in structure to Class 1 compounds, except that the substituents on the right aromatic ring are different. Class 2 compounds are advantageous in being used as AR antagonists and as therapeutic agents for hormone-refractory prostate cancer. They can be used to treat other AR-related diseases or conditions such as benign prostatic hyperplasia, alopecia or acne. These compounds and related compounds can also be used as modulators of other nuclear receptors such as estrogen receptors and peroxisome proliferator-activated receptors, as well as therapeutic agents for diseases such as breast cancer, ovarian cancer, diabetes, heart disease and metabolic-related diseases that act as nuclear receptors. They are useful in tests, for example, as standards or intermediates or prodrugs.

[0826] Table 6

[0827]

[0828]

[0829] Category 3

[0830] Class 3 compounds (see Table 7) performed slightly better than bicalutamide in treating prostate cancer. RD133, RD134, and RD138 (in Class 2) caused a greater reduction in PSA levels in the LNCaP-AR cell line than RD135 and RD137 (in Class 3). All of these compounds caused a greater reduction in PSA levels than bicalutamide.

[0831] The other three classes of compounds (not shown) are not diarylthiohydantoins and their activities are comparable to the prior art monoarylhydantoin compounds RD2, RD4 and RD5.

[0832] Therefore, 3 types of compounds are used as AR antagonists, and as therapeutic agents for hormone-refractory prostate cancer. They can also be used to treat other AR related diseases or conditions such as benign prostatic hyperplasia, alopecia or acne. These compounds and related compounds can also be used as modulators of other nuclear receptors such as estrogen receptors and peroxisome proliferator-activated receptors, and as therapeutic agents for diseases such as breast cancer, ovarian cancer, diabetes, heart disease and metabolic-related diseases that act as nuclear receptors. They are useful, for example, as standards or intermediates or prodrugs in tests.

[0833] Table 7

[0834]

[0835] Category 4

[0836] Class 4 compounds (see Table 8) were judged to be no better than bicalutamide for treating prostate cancer. For example, RD39 and RD40 from Class 4 differ from RD37 from Class 1 only in the substituents on the lower right carbon of the hydantoin ring. Substituents on the right aromatic ring may also affect activity.

[0837] Some of the Class 4 compounds (including those shown and those not shown) are not diaryl compounds (no right-hand aromatic ring), are not thiohydantoins, have no substituents on the lower right carbon of the hydantoin ring, and / or have substituents other than oxygen and amide groups on the lower left carbon of the hydantoin ring. This provides evidence of the surprising advantages of diarylthiohydantoins that are substituted on the lower right carbon of the hydantoin ring and that have oxygen or amide groups on the lower left carbon of the hydantoin ring.

[0838] Therefore, the four classes of compounds can be used as AR antagonists, and therapeutic agents for hormone-refractory prostate cancer, at least to a certain extent, they are comparable to bicalutamide. They can be used to treat other AR-related diseases or conditions such as benign prostatic hyperplasia, alopecia and acne. These and related compounds can also be used as modulators of other nuclear receptors such as estrogen receptors and peroxisome proliferator-activated receptors, and therapeutic agents for diseases such as breast cancer, ovarian cancer, diabetes, heart disease and metabolic-related diseases that act as nuclear receptors. They are useful, for example, as standards or intermediates or prodrugs in tests.

[0839] Table 8

[0840]

[0841]

[0842] 5 categories

[0843] Class 5 compounds (see Table 9) are inactive or nearly inactive and are therefore worse than bicalutamide in treating prostate cancer. The substituents on the right aromatic ring are important in determining activity.

[0844] Some of the five classes of compounds (some of which are shown and some are not) are not diaryl compounds (no right-hand aromatic ring), are not thiohydantoins, have no substituents on the lower right carbon of the hydantoin ring, and / or have substituents other than oxygen or amide groups on the lower left carbon of the hydantoin ring. This provides evidence of the surprising advantages of diarylthiohydantoins that have substituents on the lower right carbon of the hydantoin ring and that have oxygen or amide groups on the lower left carbon of the hydantoin ring. In particular, in RD155, RD156, and 158 (CH2NR x Ry , where R x,y =H or methyl) are not believed to contribute to the activity of these compounds.

[0845] Class 5 compounds are not desirable for treating prostate cancer or as AR antagonists, although these and related compounds may be useful as modulators of other nuclear receptors such as estrogen receptors and peroxisome proliferator-activated receptors, and as therapeutic agents for diseases in which nuclear receptors play a role, such as breast cancer, ovarian cancer, diabetes, heart disease, and metabolic-related diseases. They are useful in testing, for example, as standards, intermediates, or prodrugs.

[0846] Table 9

[0847]

[0848] 6 categories

[0849] 6 class compounds (referring to Table 10) are inactive or close to inactive, and are strong agonists, so they are worse than bicalutamide in the treatment of prostate cancer.These comparison compounds are classified very low with respect to the compounds of this invention.Significantly, RD72 has poor activity, and it has a chlorine substituent on the aromatic ring on the left, yet has the RD7 of trifluoromethyl, and has the RD100 of iodine, is classified in 1 class.The result for 6 class compounds provides the evidence of the surprising advantage of diaryl thiohydantoin, this diaryl thiohydantoin compound has a substituent on the lower right carbon of hydantoin ring, has oxygen or amide groups on the lower left carbon of hydantoin ring, and has definite substituent on the aromatic ring on the left.

[0850] Class 6 compounds cannot be used to treat prostate cancer or as AR antagonists.

[0851] Table 10

[0852]

[0853] Unclassified compounds

[0854] For some compounds, there was insufficient experimental data to classify them. These unclassified compounds are shown in Table 11.

[0855] Based on the data and methods of the present invention, and applying the analysis of many compounds, including those not shown here, some observations of these unclassified compounds can be obtained. The example RD1 of contrast is expected to be in Class 3 with the example RD3-RD5 of contrast. RD89 is expected to be hydrolyzed to RD37 (Class 1), so they should have similar activity. RD104 is expected to be hydrolyzed to RD58 (Class 1), so they should have similar activity. RD105 is expected to be hydrolyzed to RD8 (Class 1), and RD 139 and RD140 are expected to be hydrolyzed to RD138 (Class 2), so they should have similar activity.

[0856] Table 11

[0857]

[0858]

[0859] In summary, novel compounds were identified and prepared that showed significant improvement over bicalutamide in the treatment of prostate cancer.

[0860] Sensitivity of anticancer activity of compounds to structural differences

[0861] The present inventors have determined that seemingly minor changes in the structure of hydantoin compounds can result in significant changes in the properties of the compounds in treating prostate cancer. For example, RD161 and RD162 differ only in a single fluorine substituent on the aromatic ring. RD162 is in Class 1, while RD161 is in Class 2. Both are better than bicalutamide in treating prostate cancer, but RD162 is better. However, RD149, which differs from RD161 only in having an additional carbon atom between the methylcarbamoyl group and the aromatic ring, is no better than bicalutamide in treating prostate cancer and is classified in Class 4. The effects of RD161, RD162, and RD149 on luciferase activity can be seen from the Figure 24 At a given compound concentration, the luciferase activity after exposure to RD161 and RD162 was less than that after exposure to RD149.

[0862] RD9 differs from RD8 only in that the amino group replaces the hydroxyl group. However, RD8 is in category 1 and is much better than bicalutamide in treating prostate cancer, while RD9 is in category 4 and is no better than bicalutamide. The effects of RD8 and RD9 on luciferase activity in the 1AR cell line can be seen from Figure 27 For a given dose, the luciferase activity of cells exposed to RD8 was lower than that of cells exposed to RD9. The effects of RD8 and RD9 on the luciferase activity in the 4AR cell line can be seen from Figure 26For a given dose, the luciferase activity of cells exposed to RD8 was lower than that of cells exposed to RD9. The effects of RD8 and RD9 on PSA levels in LN / AR cell lines can be seen from Figure 25 For a given dose, PSA levels were lower in patients exposed to RD8 than in patients exposed to RD9.

[0863] RD130 and RD131 differ from each other only by the methyl substituent at the end of the carbamoyl group, and both compounds are classified in Class 1, although RD131 has been found to be particularly advantageous. RD129 is identical to RD130, except that the methoxy group replaces the amino group. However, RD129 is classified in Class 3. RD128 is similar to RD129, but has one less carbon in the ester chain attached to the aromatic ring, and RD128 is classified in Class 3. The effects of RD130, RD131, RD128, and RD129 on PSA levels in LN / AR cell lines can be seen in Table 28. For a given concentration, PSA levels after exposure to RD130 and RD131 were less than those after exposure to RD128 and RD129.

[0864] RD153 and RD155 differ from each other in that the former has a methylcarbamoyl group attached to the aromatic ring and a dimethyl substituent attached to the thiohydantoin group, while the latter has a methylamino group attached to the right aromatic ring and a cyclobutyl substituent attached to the thiohydantoin group. However, RD153, in Category 1, is significantly more effective than bicalutamide in treating prostate cancer, while RD155, in Category 5, is inactive or nearly inactive in treating prostate cancer. The effects of RD153 and RD155 on luciferase activity in the LN / AR cell line can be seen in Table 29. For a given concentration, luciferase activity following exposure to RD153 is less than that following exposure to RD155.

[0865] RD58 and RD60 differ from each other in the substitution of thio and oxo groups and the substitution of dimethyl and cyclobutyl substituents. RD58 is in class 1 and RD60 is in class 4.

[0866] Pharmaceutical compositions and administration

[0867] The compounds of the present invention may be used in pharmaceutical compositions prepared using a therapeutically effective amount of a compound of the present invention as defined herein, and a pharmaceutically acceptable carrier or diluent.

[0868] The diarylhydantoin compounds of the present invention can be formulated into pharmaceutical compositions and administered to a patient in need of treatment, e.g., a mammal such as a human patient, in a variety of forms suitable for the selected route of administration, e.g., orally, intranasally, intraperitoneally, or parenterally, by intravenous, intramuscular, topical or subcutaneous routes, or by injection into a tissue.

[0869] Thus, the diarylhydantoin compounds of the present invention can be administered systemically, such as orally or by inhalation or insufflation, in combination with a pharmaceutically acceptable carrier, such as an inert diluent or a concurrently edible carrier. They can be enclosed in hard-shell or soft-shell gelatin capsules, compressed into tablets, or incorporated directly into the patient's diet. For oral therapeutic administration, the diarylhydantoin compounds can be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, and the like. The diarylhydantoin compounds can be combined with a fine inert powder carrier and inhaled or insufflated by the patient. Such compositions and formulations should contain at least 0.1% of the diarylhydantoin compound. Of course, the percentage in such compositions and formulations can vary, but generally can range from about 2% to about 60% by weight of a given unit dosage form. The amount of the diarylhydantoin compound in such therapeutically useful compositions is such that an effective dosage level is obtained.

[0870] The tablets, lozenges, pills, capsules, etc. may also contain the following additives: binders such as gum tragacanth, gum arabic, corn starch or gelatin, diluents such as dicalcium phosphate, disintegrants such as corn starch, potato starch, alginic acid, etc., lubricants such as magnesium stearate, and sweeteners such as sucrose, fructose, lactose or aspartame, or flavorings such as mint, wintergreen oil, or cherry flavoring. When the unit dosage form is a capsule, in addition to the above-mentioned types of substances, it may contain a liquid carrier such as vegetable oil or polyethylene glycol. Various other substances may exist as coating agents or in other modified physical forms of solid unit dosage forms. For example, tablets, pills or capsules can be coated with gelatin, paraffin, shellac or sucrose, etc. Syrups or elixirs may contain the active compound, sucrose or fructose as sweeteners, methylparaben and propylparaben as preservatives, dyes and flavorings such as cherry or orange flavor. Of course, any substance used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amount used. In addition, the diarylhydantoin compound can be incorporated into a timed-release formulation and carrier. For example, the diarylhydantoin compound can be incorporated into a timed-release capsule, a timed-release tablet, and a timed-release pill.

[0871] The diaryl hydantoin compound can be administered intravenously or intraperitoneally by infusion or injection. The solution of the diaryl hydantoin compound can be prepared with water, optionally mixed with a nontoxic surfactant. Dispersion liquids can also be prepared with glycerol, liquid polyethylene glycol, triacetin and mixtures thereof and with oil. Under the usual circumstances of preservation and use, these preparations can contain preservatives to stop the growth of microorganisms.

[0872] The pharmaceutical dosage form suitable for injection or infusion may include a sterile aqueous solution or dispersion or sterile powder containing the diarylhydantoin compound, which is suitable for extemporaneous preparations of sterile injection or infusion solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or excipient may be a solvent or liquid dispersion medium, comprising, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), a vegetable oil, a non-toxic glyceride, i.e., a suitable mixture thereof. Suitable fluidity can be maintained by the following methods, for example, by forming liposomes, by maintaining the desired particle size in the case of a dispersion, or by using a surfactant. Preventing the action of microorganisms can be achieved by different antibacterial and antifungal agents, for example, parabens, gem-chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it is preferably comprised of an isotonic agent, for example, a sugar, a buffer or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0873] Sterile injectable solutions can be prepared by introducing the diarylhydantoin compound in the required amount in a suitable solvent and, if desired, adding the other ingredients listed above, followed by sterile filtration. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying techniques, which produce a powder of the active ingredient plus any other desired ingredients present in a previously sterile-filtered solution.

[0874] For topical administration, the diarylhydantoin compounds can be applied in pure form. However, they will generally be administered to the skin as compositions or formulations in combination with a dermatologically acceptable carrier, which can be a solid or a liquid.

[0875] Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silicon dioxide, alumina, etc. Other solid carriers include nontoxic polymer nanoparticles or microparticles. Useful liquid carriers include water, alcohols or ethylene glycol or water / alcohol / ethylene glycol mixtures, in which the diarylhydantoin compound can be dissolved or dispersed at an effective level, optionally with auxiliary nontoxic surfactants. For given purposes, auxiliary agents such as aromatic substances and other antimicrobial agents can be added to optimize their properties. The resulting liquid composition can be applied to absorbent pads, impregnated bandages and other dressings, or sprayed onto the active area using a pump or aerosol sprayer.

[0876] For direct application to the user's skin, thickening agents such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified minerals and liquid carriers may also be used to form easily spreadable pastes, gels, ointments, soaps, etc.

[0877] Examples of useful dermatological compositions that can be used to deliver diarylhydantoin compounds to the skin are known in the art, for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157), and Wortzman (U.S. Pat. No. 4,820,508), all of which are incorporated herein by reference.

[0878] The useful dosage of the compound of formula I can be determined by comparing their in vitro activity and in vivo activity in animal models. Methods for deducing the dosage used in humans from the effective dose in mice and other animals are known in the art, for example, referring to U.S. Patent No. 4,938,949, which is incorporated herein by reference.

[0879] For example, the concentration of the diarylhydantoin compound in liquid compositions such as lotions may be about 0.1-25% by weight or about 0.5-10% by weight. The concentration in semisolid or solid compositions such as gels or powders may be about 0.1-5% by weight or about 0.5-2.5% by weight.

[0880] The amount of diarylhydantoin compound required for use in therapy will vary not only with the particular salt selected, but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient, and is ultimately at the discretion of the attending physician or clinician.

[0881] The effective dose and route of administration of the medicament of the present invention are conventional doses and routes. The precise amount of the medicament (effective dose) will vary with the patient's, for example, race, age, weight and general or clinical condition of the patient, the severity or mechanism of any disease to be treated, the special agent or tool used, the method and arrangement of administration, etc. The therapeutically effective dose can be determined empirically by conventional methods known to those skilled in the art. See, for example, The Pharmacological Basis of Therapeutics, Goodman and Gilman, eds., Macmillan Publishing Co., New York. For example, the effective dose can be initially determined in cell culture experiments or in suitable animal models. This animal model can also be used to determine a suitable dosage concentration range and route. This information can be used to determine useful doses and routes when administering to humans. The therapeutic dose can also be selected by analogy with the dosage of similar therapeutic agents.

[0882] The particular mode of administration and dosage regimen will be chosen by the attending physician, taking into account the particulars of the situation (e.g., the patient, the disease, the stage of the condition involved, and whether the treatment is prophylactic). Treatment may involve daily or multiple daily doses of the compound administered over a period of several days to several months or even years.

[0883] In general, suitable dosages will be in the range of from about 0.001 to about 100 mg / kg, e.g., from about 0.01 to about 100 mg per kilogram of body weight per day, such as above about 0.1 mg per kilogram, or in the range of about 1 to about 10 mg per kilogram of recipient body weight per day, for example, a suitable dosage may be about 1 mg / kg, 10 mg / kg or 50 mg / kg of body weight per day.

[0884] The diarylhydantoin compound may conveniently be administered in unit dosage form, for example, containing 0.05 to 10,000 mg, 0.5 to 10,000 mg, 5 to 1,000 mg, or about 100 mg of active ingredient per unit dosage.

[0885] The diarylhydantoin compound can be administered to achieve peak plasma concentrations, for example, from about 0.5 to about 75 μM, about 1 to 50 μM, about 2 to about 30 μM, or about 5 to about 25 μM. Representative desired plasma concentrations include at least or no more than 0.25, 0.5, 1, 5, 10, 25, 50, 75, 100, or 200 μM. For example, plasma levels can be from about 1 to 100 micromolar or from about 10 to about 25 micromolar. This can be achieved, for example, by intravenous injection of a 0.05-5% solution of the diarylhydantoin compound, optionally in saline, or orally as a bolus containing about 1-100 mg of the diarylhydantoin compound. The desired blood levels can be maintained by continuous infusion to provide about 0.00005-5 mg per kilogram of body weight per hour, for example, at least or no more than 0.00005, 0.0005, 0.005, 0.05, 0.5, or 5 mg / kg / hr. Alternatively, such levels can be achieved by periodic infusions containing about 0.0002-20 mg per kilogram of body weight, for example, at least or no more than 0.0002, 0.002, 0.02, 0.2, 2, 20, or 50 mg of the diarylhydantoin compound per kilogram of body weight.

[0886] The diarylhydantoin compound may conveniently be presented in a single dose or as divided doses to be administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-doses themselves may be further divided, for example, into a number of discrete, loosely spaced administrations, such as a number of inhalations from an insufflator.

[0887] Many of the above-mentioned compounds determined show little or no agonist activity against hormone-refractory prostate cancer cells. Because these compounds are strong AR inhibitors, they can be used not only to treat prostate cancer, but also to treat other AR-related diseases or conditions such as benign prostatic hyperplasia, alopecia and acne. Since AR belongs to the nuclear receptor family, these compounds can be used as drug synthesis to guide the skeleton (scaffold) of other nuclear receptors, such as estrogen receptors and peroxisome proliferator-activated receptors. Therefore, they can be further developed for other diseases such as breast cancer, ovarian cancer, diabetes, heart disease and metabolic-related diseases in which nuclear receptors work.

[0888] The embodiments shown and discussed in this specification are intended only to guide those skilled in the art to best understand the present invention and thereby implement and utilize the present invention. Nothing in this specification should be construed as limiting the scope of the present invention. All examples provided are representative and non-limiting. The embodiments described above in the present invention may be modified and varied as those skilled in the art may make in light of the above teachings without departing from the present invention. It is therefore understood that the present invention may be implemented in other than the specifically described embodiments within the scope of the claims and their equivalents.

[0889] The present invention also relates to the following items:

[0890] 1. A compound having the formula:

[0891]

[0892] wherein X is selected from trifluoromethyl and iodine,

[0893] wherein W is selected from O and NR5,

[0894] wherein R5 is selected from H, methyl and

[0895]

[0896] wherein D is S or O, and E is N or O, and G is alkyl, aryl, substituted alkyl, or substituted aryl; or

[0897] D is S or O and EG together are C1-C4 lower alkyl,

[0898] wherein R1 and R2 together comprise 8 or fewer carbon atoms and are selected from alkyl, substituted alkyl including haloalkyl, and cycloalkyl or substituted cycloalkyl together with the carbon to which they are attached,

[0899] wherein R is selected from the group consisting of hydrogen, halogen, methyl, C1-C4 alkoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methoxycarbonyl, acetylamino, methylsulfonylamino, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, and C1-C6 alkyl or alkenyl, optionally substituted with hydroxy, methoxycarbonyl, cyano, amino, amido, nitro, carbamoyl, or substituted carbamoyl including methylcarbamoyl, dimethylcarbamoyl and hydroxyethylcarbamoyl,

[0900] wherein R4 is selected from the group consisting of hydrogen, halogen, alkyl and haloalkyl,

[0901] Wherein R3 is not methylaminomethyl or dimethylaminomethyl.

[0902] 2. The compound of item 1, wherein R5 is

[0903]

[0904] 3. The compound of item 1, which has the following formula:

[0905]

[0906] wherein R3 is selected from hydroxy, methylcarbamoyl, methylcarbamoylpropyl, methylcarbamoylethyl, methylcarbamoylmethyl, methylsulfonylcarbamoylpropyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, carbamoylmethyl, carbamoylethyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoylpropyl, carboxypropyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, hydroxyethylcarbamoylethyl and hydroxyethoxycarbonylethyl, and

[0907] wherein R10 and R11 are both H, or are F and H, or H and F, respectively.

[0908] 4. The compound of item 3, wherein R10 and R11 are both H.

[0909] 5. The compound of item 3, wherein R10 and R11 are F and H respectively.

[0910] 6. The compound of item 3, wherein R3 is methylcarbamoyl.

[0911] 7. The compound of item 3, wherein R3 is methylcarbamoyl and R10 and R11 are F and H, respectively.

[0912] 8. The compound of item 1,

[0913] wherein R1 and R2 are independently methyl, or together with the carbon to which they are attached, are cycloalkyl having 4-5 carbon atoms, and

[0914] R3 is selected from carbamoyl, alkylcarbamoyl, carbamoylalkyl and alkylcarbamoylalkyl, and R4 is H or F.

[0915] 9. The compound of item 8, wherein R4 is 3-fluoro.

[0916] 10. The compound of item 1,

[0917] wherein R1 and R2 are independently methyl, or a cycloalkyl group having 4-5 carbon atoms together with the carbon atoms to which they are attached,

[0918] R3 is selected from cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-(1,1-dimethylethoxycarbonyl)-1-piperazinyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxyethoxycarbonyl-substituted alkyl and 3-cyano-4-trifluoromethylphenylcarbamoyl, and R4 is F.

[0919] 11. The compound of item 1, which has the following formula:

[0920]

[0921] wherein R3 is selected from methylcarbonyl, methoxycarbonyl, acetylamino and methylsulfonylamino, and R4 is selected from F and H.

[0922] 12. The compound of claim 1, which has the formula:

[0923]

[0924] wherein R4 is selected from F and H.

[0925] 13. The compound of claim 1, wherein R1 and R2 together with the carbon to which they are attached are:

[0926]

[0927] 14. A compound selected from compounds of Class 1 and Class 2.

[0928] 15. The compound of item 1, which has the following formula:

[0929]

[0930] 16. The compound of item 1, which has the following formula:

[0931]

[0932] 17. The compound of item 1, which has the following formula:

[0933]

[0934] 18. The compound of item 1, which has the following formula:

[0935]

[0936] 19. The compound of claim 1, which has the formula:

[0937]

[0938] 20. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of items 1 to 19 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0939] 21. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to item 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0940] 22. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to item 9 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

[0941] 23. A method for treating a hyperproliferative disease, said method comprising administering the pharmaceutical composition of item 20 to a patient in need of such treatment, thereby treating the hyperproliferative disease.

[0942] 24. The method of claim 23, wherein the composition is administered at a dose of the compound ranging from about 0.001 mg per kilogram of body weight per day to about 100 mg per kilogram of body weight per day.

[0943] 25. The method of claim 23, wherein the composition is administered at a dose of the compound ranging from about 0.01 mg per kilogram of body weight per day to about 100 mg per kilogram of body weight per day.

[0944] 26. The method of claim 23, wherein the composition is administered at a dose of the compound ranging from about 0.1 mg per kilogram of body weight per day to about 10 mg per kilogram of body weight per day.

[0945] 27. The method of claim 23, wherein the composition is administered at a dose of about 1 mg of the compound per kilogram of body weight per day.

[0946] 28. A method for treating a hyperproliferative disease, said method comprising administering the composition of item 21 to a patient in need of such treatment, thereby treating the hyperproliferative disease.

[0947] 29. The method of claim 28, wherein the composition is administered at a dose of the compound ranging from about 0.1 mg per kilogram of body weight per day to about 10 mg per kilogram of body weight per day.

[0948] 30. The method of claim 28, wherein the composition is administered at a dose of about 1 mg of the compound per kilogram of body weight per day.

[0949] 31. The method of claim 23, wherein the hyperproliferative disease is hormone-refractory prostate cancer.

[0950] 32. The method of claim 23, wherein the compound is administered intravenously, by injection into a tissue, intraperitoneally, orally, or intranasally.

[0951] 33. The method of claim 28, wherein the composition is administered orally.

[0952] 34. The method of claim 23, wherein the composition has a form selected from the group consisting of a solution, a dispersion, a suspension, a powder, a capsule, a tablet, a pill, a timed-release capsule, a timed-release tablet, and a timed-release pill.

[0953] 35. The method of claim 28, wherein the composition has a form selected from the group consisting of a capsule, a tablet, and a pill.

[0954] 36. The method of claim 28, wherein the compound is selected from RD162', RD162", RD169 or RD170, or a pharmaceutically acceptable salt thereof.

[0955] 37. The method of claim 28, wherein the compound is N-methyl-4-[7-(4-cyano-3-trifluoromethylphenyl)-8-oxo-6-thioxo-5,7-diaza-spiro[3.4]octan-5-yl]-2-fluorobenzamide [RD162] or a pharmaceutically acceptable salt thereof.

[0956] 38. A method for synthesizing a diaryl compound of the following formula:

[0957]

[0958] The method comprises mixing compound I and compound II in a first polar solvent to form a mixture;

[0959]

[0960] Heat the mixture,

[0961] A second polar solvent, which may be the same as or different from the first polar solvent, and an aqueous solution of an acid are added to the mixture,

[0962] The mixture was refluxed,

[0963] The mixture is cooled and mixed with water, and

[0964] separating the diaryl compound from the mixture,

[0965] wherein R51 comprises an alkyl chain of 1 to 4 carbon atoms, R52 is selected from cyano, hydroxy, methylcarbamoyl, methylcarbamoyl-substituted alkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, 3-cyano-4-trifluoromethylphenylcarbamoyl, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl and hydroxyethoxycarbonyl-substituted alkyl, and R53 is selected from F and H.

[0966] 39. The method of claim 38, wherein R51 comprises an alkyl chain of 1-2 carbon atoms, R52 is selected from carbamoyl and methylcarbamoyl, and R53 is F.

[0967] 40. A method for synthesizing the compound of the following formula:

[0968]

[0969] The method comprises

[0970] mixing 4-isothiocyanato-2-trifluoromethylbenzonitrile and N-methyl-4-(1-cyanocyclobutylamino)-2-fluorobenzamide in dimethylformamide to form a first mixture,

[0971] heating the first mixture to form a second mixture,

[0972] To this second mixture is added an alcohol and an acid to form a third mixture,

[0973] refluxing the third mixture to form a fourth mixture,

[0974] cooling the fourth mixture,

[0975] The fourth mixture was mixed with water and the organic layer was extracted,

[0976] The compound was isolated from the organic layer.

[0977] 41. A method for synthesizing the compound [RD162'] of item 16, comprising

[0978] combining N-methyl-2-fluoro-4-(1,1-dimethyl-cyanomethyl)-aminobenzamide and 4-isothiocyanato-2-trifluoromethylbenzonitrile in DMF and heating to form a first mixture;

[0979] adding an alcohol and an acid to this first mixture to form a second mixture;

[0980] reflux the second mixture;

[0981] Cooling the second mixture,

[0982] mixing the second mixture with water and extracting the organic layer;

[0983] From the organic layer, the mixture was separated.

[0984] 42. A method for synthesizing the compound [RD162"] of item 17, comprising

[0985] mixing N-methyl-2-fluoro-4-(1-cyanocyclopentyl)aminobenzamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile, and DMF and heating to reflux to form a first mixture;

[0986] adding an alcohol and an acid to the first mixture to form a second mixture;

[0987] reflux the second mixture;

[0988] Cooling the second mixture;

[0989] The second mixture is mixed with water and the organic layer is extracted;

[0990] The compound was isolated from the organic layer.

[0991] 43. A method for synthesizing the compound [RD169] of item 18, the method comprising

[0992] mixing N,N-dimethyl-4-[4-(1-cyanocyclobutylamino)phenyl]butanamide, 4-isothiocyanato-2-trifluoromethylbenzonitrile, and DMF and heating to reflux to form a first mixture;

[0993] adding an alcohol and an acid to this first mixture to form a second mixture;

[0994] reflux the second mixture;

[0995] Cooling the second mixture;

[0996] The second mixture is mixed with water and the organic layer is extracted;

[0997] The compound was isolated from the organic layer.

[0998] 44. A method for synthesizing the compound [RD170] of item 19, comprising

[0999] Mix DMSO, dichloromethane and oxalyl chloride to form a first mixture,

[1000] To this first mixture was added 4-(4-(7-(4-cyano-3-(trifluoromethyl)phenyl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]octan-5-yl)phenyl)butanamide to form a second mixture;

[1001] adding triethylamine to the second mixture to form a third mixture;

[1002] The third mixture was warmed and quenched with aqueous NH4Cl to form a fourth mixture;

[1003] extracting an organic layer from the fourth mixture;

[1004] The compound was isolated from the organic layer.

[1005] 45. Compounds of the formula:

[1006]

[1007] Where R5 is CN or NO2 or SO2R 11 ,

[1008] wherein R6 is CF3, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, halogen,

[1009] Wherein A is sulfur (S) or oxygen (O),

[1010] Wherein B is O or S or NR8,

[1011] wherein R8 is selected from H, methyl, aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, SO2R 11 NR 11 R 12 、(CO)OR 11 、(CO)NR 11 R 12 、(CO)R 11 、(CS)R 11 、(CS)NR 11 R 12 、(CS)OR 11 、

[1012]

[1013] wherein D is S or O, and E is N or O, and G is alkyl, aryl, substituted alkyl, or substituted aryl; or

[1014] D is S or O and EG together are C1-C4 lower alkyl,

[1015] wherein R1 and R2 are independently alkyl, haloalkyl, hydrogen, aryl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic heterocyclic group or non-aromatic heterocyclic group, substituted aromatic heterocyclic group or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, or R1 and R2 are linked to form a ring, which can be a heterocyclic group, a substituted heterocyclic group, a cycloalkyl, a substituted cycloalkyl,

[1016]

[1017] wherein X is carbon or nitrogen and may be at any position on the ring, and

[1018] wherein R3, R4 and R7 are independently selected from hydrogen, halogen, methyl, methoxy, formyl, haloacetoxy, trifluoromethyl, cyano, nitro, hydroxy, phenyl, amino, methylcarbamoyl, methylcarbamoyl-substituted alkyl, dimethylcarbamoyl-substituted alkyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, methylsulfonyl, 4-methylsulfonyl-1-piperazinyl, piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxy-substituted alkyl, hydroxy-substituted alkenyl, carbamoyl-substituted alkenyl, methoxycarbonyl-substituted alkyl, cyano-substituted alkyl, Aryl, substituted aryl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkenyl, haloalkynyl, SO2R 11 NR 11 R 12 NR 12 (CO)OR 11 、NH(CO)NR 11 R 12 NR 12 (CO)R 11 、O(CO)R 11 、O(CO)OR 11 、O(CS)R 11 NR 12 (CS)R 11 、NH(CS)NR 11 R 12 NR 12 (CS)OR 11, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, substituted cycloalkyl, haloalkyl, methylsulfonylcarbamoyl-substituted alkyl, methylaminomethyl, dimethylaminomethyl, methylsulfonyloxymethyl, methoxycarbonyl, acetylamino, methylsulfonylamino, carbamoyl-substituted alkyl, carboxymethyl, methoxycarbonylmethyl, methylsulfonyl, 4-cyano-3-trifluoromethylphenylcarbamoyl-substituted alkyl, carboxy-substituted alkyl, 4-(1,1-dimethylethoxy)carbonyl)-1-piperazinyl, hydroxyethylcarbamoyl-substituted alkyl, hydroxyethoxycarbonyl-substituted alkyl, 3-cyano-4-trifluoromethylphenylcarbamoyl,

[1019] where R 11 and R 12 are independently hydrogen, aryl, aralkyl, substituted aralkyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, haloalkyl, haloalkenyl, haloalkynyl, arylalkyl, arylalkenyl, arylalkynyl, aromatic or non-aromatic heterocyclic group, substituted aromatic or non-aromatic heterocyclic group, cycloalkyl, or substituted cycloalkyl, or R 11 and R 12 They may be linked to form a ring, which may be an aromatic heterocyclic group or a non-aromatic heterocyclic group, a substituted aromatic heterocyclic group, a cycloalkyl group, or a substituted cycloalkyl group.

[1020] 46. ​​The compound of claim 45, wherein the compound has significant androgen receptor antagonist activity and substantially no agonist activity against hormone-refractory prostate cancer cells.

[1021] 47. A method comprising:

[1022] Providing at least one compound according to item 45;

[1023] measuring inhibition of androgen receptor activity by the compound and determining whether the inhibition is above a first predetermined level,

[1024] measuring stimulation of androgen receptor activity in hormone-resistant cancer cells by the compound and determining whether the stimulation is below a second predetermined level,

[1025] If the inhibition is above a first predetermined level and the stimulation is below a second predetermined level, the compound is selected.

[1026] 48. The method of claim 47, wherein the predetermined levels are those of bicalutamide.

[1027] 49. The method of claim 47, wherein the measuring inhibition step comprises measuring inhibitory concentration (IC50) in an AR-responsive reporter system or a prostate-specific antigen secretion system.

[1028] 50. The method of item 47, wherein the step of measuring stimulation comprises measuring the fold induction by increasing concentrations in an AR-responsive reporter system or a prostate-specific antigen secretion system.

[1029] 51. The method of claim 47, wherein said measuring inhibition and / or stimulation step comprises measuring the effect of said compound on tumor growth in an animal.

Claims

1. A compound having the formula: or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. The pharmaceutical composition of claim 2, wherein the pharmaceutically acceptable carrier is a pharmaceutically acceptable diluent.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating prostate cancer.

5. Use of a compound according to claim 1 for treating prostate cancer in a subject.

6. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating Cushing's disease.

7. Use of a compound according to claim 1 for treating Cushing's disease in a subject.

8. Compounds of the formula: (tert-Butyl 4-(4-aminophenyl)-piperazine-1-carboxylate).

9. Compounds of the formula:

10. A method for testing the antagonistic activity of a compound in the presence of R1881, comprising the following steps: A panel of genetically engineered LNCaP cells containing an androgen receptor (AR)-responsive reporter (LNCaP-AR) was maintained in Iscove's medium containing 10% fetal bovine serum (FBS); This group of cells was grown in Iscove's medium containing 10% charcoal-stripped FBS (CS-FBS) to deprive of androgens; A group of test cells is split from the group of cells; Growth in Iscove's medium containing 10% CS-FBS, 100 pM R1881, and several different concentrations of compounds; Cultivating the cells for 2 days; and Detect reporter activity.

11. An HPLC analysis method comprising the following steps: providing a blood sample containing medication; Blood cells are separated from the serum of a blood sample by centrifugation; To 400 μl of serum, 80 μl of 10 μM internal standard solution was added to form a mixture; 520 μl of acetonitrile was added to the mixture to induce precipitation; This mixture was vortexed and then sonicated; separating the solid particles by filtration; The filtrate was dried to dryness under a stream of argon; Acetonitrile was added to the dried filtrate to form 80 μL of HPLC sample; and The samples were analyzed by HPLC using a standard curve of the drug to determine the drug concentration.

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