Synthesis of novel imipridone derivatives and evaluation of their anticancer activity

Abstract

The present invention relates to a compound of formula (I), or a pharmaceutically acceptable salt and stereoisomer thereof (including an enantiomer, a racemic mixture, a mixture of enantiomers, or a combination thereof), which is applicable for use in the treatment of cancer diseases. The present invention further relates to a pharmaceutical composition comprising said compound:

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a compound of formula (I) [ka] or a pharmaceutically acceptable salt and stereoisomer thereof (including a single enantiomer, a racemic mixture, a mixture of enantiomers, or a combination thereof), which are applicable for use in the treatment of cancer diseases. The present invention further relates to a pharmaceutical composition comprising said compound. [Background technology]

[0002] Background of the Invention Imipridone, a first-in-class small molecule anticancer compound, contains an angular tricyclic heterocyclic framework enhanced in basicity by two substituted backbone nitrogen atoms [see the general structure represented by Formula (I)]. This well-positioned, ideal number of basic centers and a lactam moiety containing adjustable aromatic rings protruding as evenly distributed potential binding sites, confer multitarget characteristics and ideal drug properties essential for their well-defined mechanism of action. Specific G protein-coupled receptors (GPCRs) that control important signaling pathways in cancer cells are key targets for ONC201 (reference compound 1), the first imipridone in clinical development that directly antagonizes a GPCR called the DRD2 dopamine receptor [1]. [ka]

[0003] This molecule has also emerged as an efficient activator of the pro-apoptotic protein tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptor, exhibiting a wide therapeutic index [2-4]. Kline et al. demonstrated that ONC201 caused dual inhibition of the AKT and extracellular signal-regulated kinase (ERK) pathways in many malignant cell lines (e.g., HCT-116, HEPG-2, MCF-7, and MDA-MB-468). They demonstrated that ONC201 induced cell cycle arrest as early as 24 hours after treatment in the cell lines tested, in addition to apoptosis as measured by sub-G1 fraction and caspase activation [2].

[0004] The aforementioned references [1-4] relate only to ONC201 and do not disclose other imipridone derivatives.

[0005] The authors also confirmed through bromodeoxyuridine (BrdU) labeling experiments that ONC201 inhibited cell proliferation and significantly reduced the number of viable cells, including those that did not undergo apoptosis (e.g., A-549 and SNV-449), due to premature cell cycle arrest within 48 hours of treatment with this imipridone [2]. Preclinical studies have demonstrated its potency as a highly promising apoptotic anticancer agent, with significant activity against a wide variety of cancer cell lines (e.g., PANC-1, HCT116, MDA-MB-23, U87, HFF, MRC5, and WI-38) [5-8]. Furthermore, phase II clinical trials have demonstrated that this compound is beneficial for the treatment of patients with a wide range of advanced malignancies [9].

[0006] Regarding the above references [5-9], it can be stated that they relate only to ONC201 and do not disclose other imipridone derivatives. In addition to ONC201, Wagner et al. disclose an isomer of ONC201, which has a linear [4,3-d] structure and is therefore outside the scope of the present invention [5]. Zhe-Zhu Jin et al. disclose the use of ONC201 in combination with AZD-8055, which is a pyrido[2,3-d]pyrimidine derivative and therefore is outside the scope of the present invention [7].

[0007] An exhaustive search for analogues

[10] identified a trifluoromethylated derivative named ONC212 (reference compound 2) as a more potent imipridone with enhanced GPCR targeting and selective involvement in tumor cell death. [ka]

[0008] This compound exhibited significantly enhanced activity at nanomolar concentrations against many different malignant cell lines, solid tumors, and hematological malignancies.

[10] Reference

[10] does not disclose such ONC201 derivatives bearing a substituted benzyl group at the N-7 position.

[0009] Importantly, ONC201 has also demonstrated improved preclinical efficacy against pancreatic cancer, melanoma, and hepatocellular carcinoma in several in vivo models, including ONC201-resistant tumors, such as the PANC-1 and cAPAN-2 human pancreatic cancer xenograft models.

[11] Reference

[11] relates only to ONC201 and its derivative ONC212 and does not disclose other imipridones. Graves et al. demonstrated that ONC201 and several related analogs are highly potent activators of ClpP.

[12] Reference

[12] does not disclose ONC201 derivatives with di- or trisubstituted benzyl groups at the N-7 position.

[0010] Given the compelling preclinical evidence of the interaction between TRAIL and redox signaling pathways involved in cancer

[17] , in our previous work

[21] , we sought to identify antiproliferative imipridones bearing ferrocene-containing substituents capable of generating reactive oxygen species (ROS). Reactive oxygen species (ROS), such as nitric oxide, superoxide anion, and other forms of free radicals [18, 19], have been shown to be involved in biological regulatory processes leading to programmed cell death (apoptosis)

[20] . Based on reference

[21] , starting with a selection of ferrocene-containing primary amines and benzylamines, a small library of ferrocene-containing derivatives and purely organic analogs (including ONC201 and ONC212, which serve as reference models without ROS-generating potential) was synthesized and tested

[21] . In vitro antiproliferative studies have shown that organometallic imipridones, especially those with two ferrocene units (e.g., 7de), exhibit significant cytotoxicity against the human malignant cell lines HT-29, HEPG2, PANC1, COLO205, A2058, and EBC1, comparable to that of ONC201, although these effects are significantly less than those exerted by ONC212. On the other hand, the effects of the organic imipridones 7ah and 7ai have proven comparable to those of ONC212

[21] .

[0011] U.S. Patent No. 10,239,877 discloses a new class of 4,7-benzyl-substituted imipridone derivatives, including a lead compound known as ONC201 and several additionally substituted analogs thereof. The disclosed compounds are potent TRAIL inducers that can be used in cancer treatment.

[0012] US Patent No. 9,376,437 discloses new substituted imipridone derivatives of the following formula (see Formula 1): [ka] wherein R1 and R2 are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic groups, and wherein when R1 represents CH2-Ph, R2 does not represent CH2-(2-CH3-Ph).

[0013] In preferred compounds, R1 is CH2-Ph and R2 is CH2-(2-Cl-Ph), R1 is CH2-Ph and R2 is CH2-(2-thienyl); R1 is CH2-Ph and R2 is CH2CH2-Ph, R1 is CH2-Ph and R2 is CH2CH2-(4-N-benzyl-piperazine), R1 is CH2-Ph and R2 is CH2-(2,4-di-F-Ph), R1 is H and R2 is CH2-(2-CH3-Ph), R1 is CH3 and R2 is CH2-(2-CH3-Ph), R1 is CH2CH2-Ph and R2 is CH2-(2-CH3-Ph).

[0014] US Patent No. 9,845,324 discloses other novel substituted imipridone derivatives of the above-referenced formula 1, wherein R1 is selected from the group consisting of H, alkyl, alkylphenyl, alkylphenyl ketone, benzylpiperazine, alkylthienyl, alkylpyridinyl, alkylisoxazolidinyl, alkylmorpholinyl, alkylthiazolyl, and alkylpyrazinyl, wherein the alkyl, alkylphenyl, alkylphenyl ketone, benzylpiperazine, alkylthienyl, alkylpyridinyl, alkylisoxazolidinyl, alkylmorpholinyl, alkylthiazolyl, and alkylpyrazinyl are optionally substituted with alkyl, alkoxyl, hydroxyl, perhalogenated alkyl, or halogen, and wherein R2 is a substituted or unsubstituted heterocycloalkylalkyl, preferably a morpholinoalkyl or piperazinylalkyl group, or wherein R2 is a substituted heteroarylalkyl, preferably a pyridylalkyl or isoxazolidinylalkyl group. The above cited documents further disclose compounds of Reference Formula 2 and Reference Formula 3 as defined therein: [ka]

[0015] US Patent Application No. 2016 / 0264574 discloses novel substituted imipridone derivatives of the above-mentioned reference formula 1, wherein R1 and R2 are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic groups, and wherein when R1 represents CH2-Ph, R2 does not represent CH2-(2-CH3-Ph).

[0016] In preferred compounds, R1 is CH2-Ph and R2 is CH2-(2-Cl-Ph), R1 is CH2-Ph and R2 is CH2-(2-thienyl); R1 is CH2-Ph and R2 is CH2CH2-Ph, R1 is CH2-Ph and R2 is CH2CH2-(4-N-benzyl-piperazine), R1 is CH2-Ph and R2 is CH2-(2,4-di-F-Ph), R1 is H and R2 is CH2-(2-CH3-Ph), R1 is CH3 and R2 is CH2-(2-CH3-Ph), R1 is CH2CH2-Ph and R2 is CH2-(2-CH3-Ph), R1 is CH2CH2NHCOOC(CH3)3 and R2 is CH2-(2-CH3-Ph), R1 is CH2CH2CH2NH2 and R2 is CH2-(2-CH3-Ph).

[0017] U.S. Patent No. 10,266,533 discloses substituted imipridone derivatives of the above formulae (Reference Formula 1, Reference Formula 2, and Reference Formula 3) [wherein in Reference Formula 1, R1 and R2 are independently selected from the group consisting of H, alkyl, alkylphenyl, alkylphenyl ketone, benzylpiperazine, alkylthienyl, alkylpyridinyl, alkylisoxazolidinyl, alkylmorpholinyl, alkylthiazolyl, and alkylpyrazinyl, wherein alkyl, alkylphenyl, alkylphenyl ketone, benzylpiperazine, alkylthienyl, alkylpyridinyl, alkylisoxazolidinyl, alkylmorpholinyl, alkylthiazolyl, and alkylpyrazinyl are optionally substituted with alkyl, alkoxy, hydroxyl, perhalogenated alkyl, or halogen, and wherein R2 is a substituted or unsubstituted heteroarylalkyl; or in Reference Formula 2, R1 is hydrogen; and R a1 , R a2 , R a3 , R a4 , and Ra5 are each independently hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m , and C(O) m wherein R m and R n are independently selected from hydrogen or C1-C4 alkyl; and X represents a halogen; or a derivative of reference formula 4. [ka] [where R a1 , R a4 , and R a5 are each hydrogen; and R a2 and R a1 are each chlorine, or R a1 , R a3 , and R a5 are each hydrogen; and where R a2 and R a4 is fluorine, or R a2 , R a4 , and R a5 are each hydrogen; and where R a1 and R a3 is chlorine, or R a2 , R a4 , and R a5 are each hydrogen; and where R a1 is methyl and R a3is fluorine, or R a2 , R a4 , and R a5 are each hydrogen; and where R a1 is fluorine and R a3 is CF3].

[0018] US Patent No. 2019 / 0194201 is a divisional application of US Patent No. 10,266,533 and discloses essentially the same substituted imipridone derivatives as its parent application.

[0019] US Patent No. 2018 / 0141946 discloses imidazole pyrimidine ketones of formula 5 [ka] where n=0 or 1; R is selected from the group consisting of H, monohalo or multihalo, C1-C6 alkyl, C1-C6 alkoxy, halo-substituted C1-C6 alkyl, nitrogen, or oxygen, and a 6-membered heterocycle having zero, one, or two heteroatom substitutions; Ar is selected from the group consisting of mono- or di-substituted aryl groups having at least one substituent selected from the group consisting of halogen, C1-C6 alkyl, and halo-substituted C1-C4 alkyl; and where, when n=1 and R is H, Ar is also not phenyl, 2-chlorophenyl, 2,4-difluorophenyl, or o-methyl-phenyl. Preferred values ​​for R are F, Cl, Br, methyl, isobutyl, methoxy, trifluoromethyl, morpholinyl, or piperazinyl. The cited reference discloses the preparation and efficacy data of 58 specific compounds.

[0020] International Patent Application No. WO2018 / 031987 discloses further substituted 4,7-dibenzyl- and 4-benzyl-7-(thiophenyl-methyl)-imipridones of formula (see formula 6): [ka] where V represents a substituted benzyl, (thiophen-2-yl)-methyl, or (thiophen-3-yl)-methyl group, and the other substituents are as defined in the cited reference. The application includes preparation and efficacy data for 27 specific compounds.

[0021] U.S. Patent Application No. 2018 / 016277 relates to novel deuterated imidazo[1,2a]pyrido[3,4-e]pyrimidin-5(1H)-one compounds, such as ONC201 and analogs. This document also discloses compositions containing the compounds of the cited document, as well as the use of such compositions, alone or in combination with other therapeutic agents, in the treatment of diseases and conditions that are beneficially treated by administering inducers of the gene encoding tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) superfamily member 10. Furthermore, the cited patent document discloses the use of imipridone derivatives in cancer treatment. Thus, the use of the compound ONC201 in the treatment of genitourinary cancer is disclosed in U.S. Patent No. 10,456,402.

[0022] US Patent No. 9,688,679 discloses the use of ONC201 in the treatment of leukemia.

[0023] International Patent Application No. WO2017 / 132661 discloses the use of compounds of the above-referenced formula 1 for treating or preventing diseases, disorders, or conditions in a subject requiring selective modulation of the activity of a class A G protein-coupled receptor (GPCR) or a class A GPCR signaling pathway, particularly for treating cancers selected from the group consisting of central nervous system tumors, brain tumors, peripheral nervous system tumors, pheochromocytoma, paraganglioma, neuroendocrine tumors, pancreatic cancer, prostate cancer, endometrial cancer, hematological malignancies, and lymphatic tumors. Suitable compounds are ONC201, 206, 212, 2013, and 236.

[0024] US Patent Nos. 10,172,862 and 10,369,154 disclose the use of the compound ONC201 for the treatment of midline glioma. Summary of the Invention [Problem to be solved by the invention]

[0025] There is a need for new compounds that have enhanced anti-cancer activity compared to the state of the art, making these compounds suitable for use in medicine. [Means for solving the problem]

[0026] Inventive discovery Through careful study of the structural features of the remaining members of the imipridone family, we have discovered that novel imipridone derivatives containing a 3,5-disubstituted benzyl group attached at the 7-position, which are relevant to the present invention, have enhanced anticancer activity. [Brief explanation of the drawings]

[0027] [Figure 1] IC50 curve generated based on the results of in vitro studies performed on the PC3 human prostate cancer cell line. [Figure 2] IC50 curve generated based on the results of in vitro studies performed on the LNCap human prostate cancer cell line. [Figure 3] IC50 curve generated based on the results of in vitro studies performed on the BxPC3 pancreatic cancer cell line. [Figure 4] IC50 curve generated based on the results of in vitro studies performed on the MiaPaCa2 pancreatic cancer cell line. [Figure 5] IC50 curve generated based on the results of in vitro studies performed on the Panc1 pancreatic cancer cell line. [Figure 6] IC50 curve generated based on the results of in vitro studies performed on the A549 lung cancer cell line. [Figure 7] IC50 curve generated based on the results of in vitro studies performed on the HCC827 lung cancer cell line. [Figure 8] IC50 curve generated based on the results of in vitro studies performed on the H1993 lung cancer cell line. [Figure 9]IC50 curve generated based on the results of in vitro studies performed on the H520 lung cancer cell line. [Figure 10] IC50 curve generated based on the results of in vitro studies performed on the MDA-MB-453 breast cancer cell line. [Figure 11] IC50 curve generated based on the results of in vitro studies performed on the MDA-MB-231 breast cancer cell line. [Figure 12] Dose-response curves obtained with compound I / 1 (ONC212) (reference) for DU145, LNCaP, and PC-3 cell lines. [Figure 13] Dose-response curves obtained with compound I / 7 (ABB-011) for DU145, LNCaP, and PC-3 cell lines. [Figure 14] Dose-response curves obtained with compound I / 3 (CZT-021) (reference) for DU145, LNCaP, and PC-3 cell lines. [Figure 15] Dose-response curves obtained with compound I / 124 (TBP-333) for DU145, LNCaP, and PC-3 cell lines. [Figure 16] Dose-response curves obtained with compound I / 6 (TBP-218) (reference) for DU145, LNCaP, and PC-3 cell lines. [Figure 17] Dose-response curves obtained with compound I / 111 (TBP-272) for DU145, LNCaP, and PC-3 cell lines. [Figure 18] Dose-response curves obtained with compound I / 149 (TBP-353) (reference) on DU145, LNCaP, and PC-3 cell lines. [Figure 19] Dose-response curves obtained with compound I / 133 (TBP-400) for DU145, LNCaP, and PC-3 cell lines. [Figure 20] Dose-response curves obtained with compound I / 30 (TBP-301) for DU145, LNCaP, and PC-3 cell lines. [Figure 21]Dose-response curves obtained with compound I / 107 (CZT-136) for DU145, LNCaP, and PC-3 cell lines. [Figure 22] Dose-response curves obtained with compound I / 1 (ONC212) (reference) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 23] Dose-response curves obtained with compound I / 7 (ABB-011) for Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 24] Dose-response curves obtained with compound I / 3 (CZT-021) (reference) for Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 25] Dose-response curves obtained with compound I / 124 (TBP-333) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 26] Dose-response curves obtained with compound I / 6 (TBP-218) (reference) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 27] Dose-response curves obtained with compound I / 111 (TBP-272) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 28] Dose-response curves obtained with compound I / 149 (TBP-353) (reference) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 29] Dose-response curves obtained with compound I / 133 (TBP-400) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 30] Dose-response curves obtained with compound I / 30 (TBP-301) on Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 31] Dose-response curves obtained with compound I / 107 (CZT-136) for Panc-1, Capan-1, and MIAPaCa-2 cell lines. [Figure 32]Dose response curves obtained with compound I / 1 (ONC212) (reference) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 33] Dose-response curves obtained with compound I / 7 (ABB-011) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 34] Dose-response curves obtained with compound I / 3 (CZT-021) (reference) for Detroit 562, SCC-25, and Fadu cell lines. [Figure 35] Dose-response curves obtained with compound I / 124 (TBP-333) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 36] Dose-response curves obtained with compound I / 6 (TBP-218) (reference) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 37] Dose-response curves obtained with compound I / 111 (TBP-272) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 38] Dose-response curves obtained with compound I / 149 (TBP-353) (reference) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 39] Dose-response curves obtained with compound I / 133 (TBP-400) on Detroit 562, SCC-25, and Fadu cell lines. [Figure 40] Dose-response curves obtained with compound I / 30 (TBP-301) in Detroit 562, SCC-25 and Fadu cell lines. [Figure 41] Dose-response curves obtained with compound I / 107 (CZT-136) for Detroit 562, SCC-25, and Fadu cell lines. [Figure 42] Dose-response curves obtained with compound I / 1 (ONC212) (reference) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 43]Dose-response curves obtained with compound I / 7 (ABB-011) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 44] Dose-response curves obtained with compound I / 3 (CZT-021) (reference) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 45] Dose-response curves obtained with compound I / 124 (TBP-333) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 46] Dose-response curves obtained with compound I / 6 (TBP-218) (reference) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 47] Dose-response curves obtained with compound I / 111 (TBP-272) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 48] Dose-response curves obtained with compound I / 149 (TBP-353) (reference) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 49] Dose-response curves obtained with compound I / 133 (TBP-400) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 50] Dose-response curves obtained with compound I / 30 (TBP-301) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 51] Dose-response curves obtained with compound I / 107 (CZT-136) for EBC-1, MDA-MB-231, and MDA-MB-453 cell lines. [Figure 52] Cytotoxic effect of compound I / 1 (ONC212) (reference) on Panc-1 cell line. [Figure 53] Cytotoxic effect of compound I / 3 (CZT-021) (reference) on Panc-1 cell line. [Figure 54]Cytotoxic effect of compound I / 6 (TBP-218) (reference) on Panc-1 cell line. [Figure 55] Cytotoxic effect of compound I / 149 (TBP-353) (reference) on Panc-1 cell line. [Figure 56] Cytotoxic effect of compound I / 30 (TBP-301) on Panc-1 cell line. [Figure 57] Cytotoxic effect of compound I / 7 (ABB-011) on Panc-1 cell line. [Figure 58] Cytotoxic effect of compound I / 124 (TBP-333) on Panc-1 cell line. [Figure 59] Cytotoxic effect of compound I / 111 (TBP-272) on Panc-1 cell line. [Figure 60] Cytotoxic effect of compound I / 133 (TBP-400) on Panc-1 cell line. [Figure 61] Cytotoxic effect of compound I / 107 (CZT-136) on Panc-1 cell line. [Figure 62] Changes in animal body weight in the experiment disclosed in Example 23. [Figure 63] Change in tumor volume in the experiment disclosed in Example 23 [*p<0.05 (Student's t-test)]. [Figure 64] Tumor size (weight) at termination in the experiment disclosed in Example 23 [*: p<0.05 (Student's t-test)]. [Figure 65] Tumors after completion of the experiment disclosed in Example 23. DETAILED DESCRIPTION OF THE INVENTION

[0028] (Brief Description of the Invention) 1. Compounds of formula (I) [ka] [where: When Y is phenyl and Z is H, X is 3-fluorophenyl-methyl (compound I / 44), 3-(aminomethyl)phenyl (compound I / 58), 4-(aminomethyl)phenyl (compound I / 62), 3-azetidinyl (compound I / 90), 4-piperidinyl (compound I / 96), 3-azidophenyl (compound I / 102), 4-azidophenyl (compound I / 104), (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl (compound I / 121), or: [ka] is; or

[0029] When Y is 3-fluorophenyl and Z is H, X is 2-iodoferrocenyl (compound I / 52), 3-(aminomethyl)phenyl (compound I / 60), 4-(aminomethyl)phenyl (compound I / 63), 3-azidophenyl (compound I / 105), or 4-azidophenyl (compound I / 106); or

[0030] When Y is 3,5-difluorophenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 61), 4-(aminomethyl)phenyl (compound I / 64), 4-azidophenyl (compound I / 107), 3-azidophenyl (compound I / 127), (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl (compound I / 132); or

[0031] When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 68); or

[0032] When Y is 3-fluoro-4-(trifluoromethyl)phenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 73); or When Y is 3-fluorophenylmethyl and Z is H, X is 2-methylphenyl (compound I / 45); or When Y is 3-(aminomethyl)phenyl and Z is H, X is 4-iodophenyl (compound I / 59), ferrocenyl (compound I / 67), 4-(trifluoromethyl)phenyl (compound I / 69), or ferrocenylmethyl (compound I / 70); or

[0033] When Y is ferrocenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 65); or When Y is ferrocenylmethyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 66); or

[0034] When Y is 3-(methoxycarbonylaminomethyl)phenyl and Z is H, X is ferrocenylmethyl (compound I / 71) or ferrocenyl (compound I / 72); or When Y is 4-aminophenyl and Z is H, X is 3-azetidinyl (compound I / 91) or 4-piperidinyl (compound I / 93); or

[0035] When Y is 3-azetidinyl and Z is H, X is 2-methylphenyl (compound I / 92); or When Y is 4-piperidinyl and Z is H, X is 2-methylphenyl (compound I / 94) or 4-fluorophenyl (compound I / 95); or When Y is 3-pyrrolidinyl and Z is H, X is 2-methylphenyl (compound I / 97) or 4-fluorophenyl (compound I / 98); or When Y is 2-pyrrolidinyl and Z is H, X is 2-methylphenyl (compound I / 99) or 4-fluorophenyl (compound I / 100); or

[0036] When Y is 4-azidophenyl and Z is H, X is 2-methylphenyl (compound I / 101); 3-fluorophenyl (compound I / 108), 4-fluorophenyl (compound I / 109), 4-(trifluoromethyl)phenyl (compound I / 110), 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 114), 4-iodophenyl (compound I / 115), 3,4,5-trimethoxyphenyl (compound I / 116), 4-azidophenyl (compound I / 117), 3-azetidinyl (compound I / 118), or 4-piperidinyl (compound I / 119); or

[0037] When Y is 3-azidophenyl and Z is H, X is 2-methylphenyl (compound I / 103), 4-(trifluoromethyl)phenyl (compound I / 111), 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 112), 4-iodophenyl (compound I / 113), or 4-chlorophenyl (compound I / 133); or When Y is (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl and Z is H, X is 2-methylphenyl (compound I / 120); or

[0038] Y is [ka] where X is 2-methylphenyl (compound I / 122); or

[0039] When Y is 3,5-diazidophenyl and Z is H, X is 4-chlorophenyl (compound I / 138) or 4-(trifluoromethyl)phenyl (compound I / 139); or When Y is 3-thiocyanatophenyl and Z is H, X is 4-chlorophenyl (compound I / 142) or 4-(trifluoromethyl)phenyl (compound I / 143); or when Y is 3-selenocyanatophenyl and Z is H, X is 4-chlorophenyl (compound I / 145) or 4-(trifluoromethyl)phenyl (compound I / 146);

[0040] or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0041] 2. The compound according to point 1, wherein When Y is 3-fluorophenyl and Z is H, X is 4-azidophenyl (compound I / 106); or When Y is 3,5-difluorophenyl and Z is H, X is 4-azidophenyl (compound I / 107); or When Y is 3-azidophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 111) or 4-chlorophenyl (compound I / 133); or when Y is 3,5-diazidophenyl and Z is H, X is 4-chlorophenyl (compound I / 138) or 4-(trifluoromethyl)phenyl (compound I / 139); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0042] 3. The compound according to point 1, wherein When Y is 3,5-difluorophenyl and Z is H, X is 4-azidophenyl (compound I / 107); or when Y is 3-azidophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 111) or 4-chlorophenyl (compound I / 133); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0043] 4. Compounds of formula (I) for use as pharmaceuticals [ka] [where: When Y is phenyl and Z is H, X is 4-fluorophenyl (compound I / 5), 3-fluorophenyl (compound I / 8), 3,4,5-trifluorophenyl (compound I / 9), 2,3,4-trifluorophenyl (compound I / 29), 2-fluoro-4-nitrophenyl (compound I / 48), or 3-aminophenyl (compound I / 75); or

[0044] When Y is 3-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 31), 4-iodophenyl (compound I / 46), 2-fluoro-4-nitrophenyl (compound I / 49), 2-methylphenyl (compound I / 53), or 4-aminophenyl (compound I / 84); or

[0045] When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-iodophenyl (compound I / 38), 4-bromophenyl (compound I / 39), 2-fluoro-4-nitrophenyl (compound I / 50), 4-chlorophenyl (compound I / 124), or 3-aminophenyl (compound I / 126), or 2-methylphenyl (compound I / 37); or

[0046] When Y is 2-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 26) or 4-iodophenyl (compound I / 54); or

[0047] When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 27); or When Y is 4-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 28) or 4-iodophenyl (compound I / 55); or

[0048] When Y is 4-aminophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 81), 4-fluorophenyl (compound I / 86), 4-iodophenyl (compound I / 87), or 3,4,5-trimethoxyphenyl (compound I / 89); or

[0049] When Y is 3,5-difluorophenyl and Z is hydroxymethyl, X is 4-chlorophenyl (racemic) (compound I / 128), 4-fluorophenyl (R-enantiomer) (compound I / 129(R)), or 4-fluorophenyl (S-enantiomer) (compound I / 129(S));

[0050] When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136) or 4-(trifluoromethyl)phenyl (compound I / 137); or

[0051] when Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11);

[0052] or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0053] 5. A compound of formula (I) for use in the treatment of cancer [ka] [where: When Y is phenyl and Z is H, X is 4-fluorophenyl (compound I / 5), 3-fluorophenyl (compound I / 8), 3,4,5-trifluorophenyl (compound I / 9), 2,3,4-trifluorophenyl (compound I / 29), 2-fluoro-4-nitrophenyl (compound I / 48), or 3-aminophenyl (compound I / 75); or

[0054] When Y is 3-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 31), 4-iodophenyl (compound I / 46), 2-fluoro-4-nitrophenyl (compound I / 49), 2-methylphenyl (compound I / 53), or 4-aminophenyl (compound I / 84); or

[0055] When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-iodophenyl (compound I / 38), 4-bromophenyl (compound I / 39), 2-fluoro-4-nitrophenyl (compound I / 50), 4-chlorophenyl (compound I / 124), or 3-aminophenyl (compound I / 126), or 2-methylphenyl (compound I / 37); or

[0056] When Y is 2-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 26) or 4-iodophenyl (compound I / 54); or

[0057] When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 27); or

[0058] When Y is 4-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 28) or 4-iodophenyl (compound I / 55); or

[0059] When Y is 4-aminophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 81), 4-fluorophenyl (compound I / 86), 4-iodophenyl (compound I / 87), or 3,4,5-trimethoxyphenyl (compound I / 89); or

[0060] When Y is 3,5-difluorophenyl and Z is hydroxymethyl, X is 4-chlorophenyl (racemic) (compound I / 128), 4-fluorophenyl (R-enantiomer) (compound I / 129(R)), or 4-fluorophenyl (S-enantiomer) (compound I / 129(S));

[0061] When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136) or 4-(trifluoromethyl)phenyl (compound I / 137); or when Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11);

[0062] or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0063] 6. The compound for use according to point 5, wherein the cancer is selected from the group consisting of prostate cancer, pancreatic cancer, lung cancer, breast cancer, glioma, head and neck cancer, colon cancer, skin cancer.

[0064] 7. Compounds for use according to points 4 to 6, wherein When Y is 3-fluorophenyl and Z is H, X is 2-methylphenyl (compound I / 53); or When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-bromophenyl (compound I / 39), 4-chlorophenyl (compound I / 124), or 2-methylphenyl (compound I / 37); or When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136), or 4-(trifluoromethyl)phenyl (compound I / 137); or when Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers or a pharmaceutically acceptable salt thereof.

[0065] 8. Compounds for use according to points 4 to 6, wherein when Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), or 4-chlorophenyl (compound I / 124); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

[0066] 9. Compounds according to points 1 to 3 for use as pharmaceuticals.

[0067] 10. Compounds according to points 1 to 3 for use in the treatment of cancer.

[0068] 11. The compound for use according to point 10, wherein the cancer is selected from the group consisting of prostate cancer, pancreatic cancer, lung cancer, breast cancer, glioma, head and neck cancer, colon cancer, skin cancer.

[0069] Detailed Description of the Invention The present invention relates to compounds of formula (I) as described in Item 1 above (wherein the identifiers in the formula are as defined in Item 1), and pharmaceutically acceptable salts thereof, and stereoisomers thereof (including single enantiomers, racemic mixtures, mixtures of enantiomers, or combinations thereof).

[0070] The group of compounds of the present invention are compounds of formula (I) above, wherein the identifiers in said formula are as defined in item 2, and pharmaceutically acceptable salts, stereoisomers thereof, including single enantiomers, racemic mixtures, mixtures of enantiomers, or combinations thereof.

[0071] Another group of compounds of the present invention are compounds of formula (I) as set forth above in item 3, wherein the identifiers in said formula are as defined in item 3, and pharmaceutically acceptable salts, stereoisomers thereof (including single enantiomers, racemic mixtures, mixtures of enantiomers, or combinations thereof).

[0072] A further group of compounds of the present invention are compounds of formula (I) as set forth above in item 4 above, wherein the identifiers in said formula are as defined in item 4, and pharmaceutically acceptable salts, stereoisomers thereof (including single enantiomers, racemic mixtures, mixtures of enantiomers, or combinations thereof).

[0073] A further group of compounds of the present invention are the compounds of formula (I) described above in item 5 above, and their pharmaceutically acceptable salts, stereoisomers (including enantiomers, racemic mixtures, mixtures of enantiomers, or combinations thereof). As described above, the compounds of the present invention exist purely or predominantly as optical isomers in the form of racemic mixtures and enantiomers. It is understood that both racemic mixtures and enantiomers, in pure form or predominantly as mixtures relative to other enantiomers, belong to the subject matter of the present invention.

[0074] The present invention also relates to a compound of the present invention for use in the treatment of a cancer disease selected from the group consisting of prostate cancer, pancreatic cancer, lung cancer, breast cancer, glioma, head and neck cancer, colon cancer, and skin cancer.

[0075] A thorough study of the structural features of the remaining members of the imipridone family described in the prior art allows us to establish that, apart from ONC234 (Reference Compound 1), besides the limited versatility of the substituents attached to N-4 and N-7, the known imipridones do not contain a di- or tri-substituted benzyl group attached to the 7-position, indicating that the accessible chemical space remains unexplored. This realization leads to the identification of novel lead compounds that may be more prominent in chemotherapeutic applications, which, in addition to enhanced anticancer activity, have additional beneficial properties, for example, in terms of a better therapeutic window and bioavailability than those in the state-of-the-art collection.

[0076] Therefore, following our convergent synthetic route shown in Reaction Scheme 1 , we carried out a diversity-oriented synthesis of novel imipridones bearing various mono-, di-, and trisubstituted benzyl groups and amine-based molecular fragments at both ends of the heterocyclic scaffold (see Table 1 ).

[0077] To expand the scope of functional groups on the pendant backbone substituents, compounds prepared from commercially available building blocks containing Boc-protected aromatic amine residues were converted to azide derivatives (see Table 1) in a one-pot procedure involving simultaneous acid-catalyzed N-deprotection and diazotization, followed by diazonium → azide exchange (see Reaction Scheme 2). Copper(I)-catalyzed coupling of the two azides with ethynylferrocene and erlotinib afforded triazole-linked hybrids (see Table 1).

[0078] During the research of the present invention, the results of biological tests revealing characteristic structure-activity relationships (SAR) were continuously taken into consideration for structural refinement in the design and synthesis of additional members of the novel imipridone family that showed enhanced cytotoxic effects. Because only a limited data set obtained from bioassays of imipridones known from the prior art (WO2018031987A1) was accessible, in addition to ONC212, three additional known halogenated analogs [I / 2 (ONC217), I / 3 (2185824-99-9P), and I / 4 (2185824-98-8P in Table 1)] were also prepared for use as references in biological tests.

[0079] After stepwise refinement, we identified three novel halogenated imipridones [I / 7 (ABB-011), I / 30 (TBP-301), and I / 39 (TBP-302)] with IC values ​​ranging from approximately 3 to 8 nM against representative cell lines. 50 We identified I / 129(R) as the most potent drug candidate, exhibiting very strong cytotoxicity, characterized by IC values, clearly demonstrating its superiority over the respective prior art reference compounds (Tables 2 and 3). We also found that the R-enantiomer of hydroxymethyl-substituted imipridone I / 129(R) [TBP-339(R)] exhibits a much stronger antiproliferative effect (IC) against the PANC1 cell line than the S-enantiomer I / 129(S) [TBP-339(S)]. 50 = 15 nM vs. 265 nM: Table 2).

[0080] The observed trend in the progressive increase in cytotoxicity produced by ONC212, 2185824-99-9P, and I / 30 (TBP-301) allows the establishment of a clear structure-activity relationship related to the stepwise introduction of fluorine substituents at the "meta" position of the N-7-benzyl group. The exact same trend is observed in the antiproliferative effects of series I / 5 (TBP-134), I / 6 (TBP-218), I / 7 (ABB-011), I / 41 (TBP-285), 2185824-98-8P, and I / 39 (TBP-302) (the most active derivative, a 3,5-difluorobenzyl-substituted derivative).

[0081] Preparation of Compounds of the Invention The convergent synthesis of novel imipridones of general formula (I), excluding compounds containing N, CN, SCN, and SeCN substituents in the side chains pendant at the 4- and 7-positions (Reaction Scheme 1), is based on direct coupling and cyclization reactions using readily available precursors such as 2-(methylthio)-4,5-dihydro-1H-imidazole (1), methyl acrylate, and primary amines of type 3 and 5. [ka]

[0082] Materials and Methods All fine chemicals were obtained from commercial sources (Merck, Fluorochem, Molar Chemicals, VWR) and used without further purification. Dioxane was distilled from sodium benzophenone. Merck Kieselgel (230-400 mesh, 60 Å) was used for flash column chromatography. Melting points (uncorrected) were determined using a Buchi M-560. All compounds 1 H NMR and 13 The C NMR spectrum was 1 H) and 125( 13 C) Recorded at MHz using a Bruker DRX-500 spectrometer in CDCl3 solution in a 5 mm tube at room temperature using the deuterium signal of the solvent as lock and TMS as internal standard. 1 H and 13 HSQC, HMBC, COSY, and NOESY spectra supporting accurate assignment of C NMR signals were obtained using standard Bruker pulse programs.

[0083] The general procedure for the synthetic steps ultimately leading to compounds of formula (I) is shown in Reaction Scheme 1. 1.2-(Methylthio)-4,5-dihydro-1H-imidazole-1-carboxylate methyl ester (2) [ka] Commercially available 2-methylthio-4,5-dihydroimidazolium iodide (12.21 g, 50 mmol) and triethylamine (TEA, 16 mL, 11.62 g, 115 mmol) were dissolved in DCM (50 mL). Methyl chloroformate (5 mL, 6.12 g, 65 mmol) was added dropwise to the solution precooled to 0 °C. The reaction mixture was warmed to 25 °C and stirred overnight. EtOAc (200 mL) was added, and after stirring for 15 min, the precipitated ammonium salt was filtered off and washed with EtOAc (50 mL). The combined solution was evaporated to dryness. The solid residue was triturated with water, filtered off, and dried under vacuum to give 5 as a white solid. Yield: 5.55 g (64%).

[0084] 2. N-substituted-4,5-dihydro-1H-imidazol-2-amine (4) [ka] To a solution of primary amine type 3 (2 mmol) dissolved in a mixture of MeOH:AcOH (4 mL:1 mL), methyl 2-(methylthio)-4,5-dihydro-1H-imidazole-1-carboxylate 2 (0.47 g, 2.4 mmol) was added, and the resulting solution was stirred and refluxed for 20 h. After cooling, the reaction was concentrated in vacuo, and the oily residue was dissolved in DCM (30 mL). The solution was washed with 3 M NaOH (10 mL), brine (10 mL), dried over NaSO, and evaporated to dryness. The colorless oil was crystallized from ether and used for cyclization to the imipridone scaffold without further purification.

[0085] 3. Imipridone-forming cyclization To the primary amine 5 (1 mmol) dissolved in MeOH (4 mL) was added methyl acrylate (0.23 mL, 2.5 mmol). The mixture was stirred at room temperature for 24 h and concentrated in vacuo. The resulting crude dipropionic acid type 6 was dissolved in anhydrous THF (4 mL). Under an argon atmosphere, NaH (0.12 g, 5 mmol) was added in small portions to the vigorously stirred solution precooled to 0 °C. The resulting suspension was stirred at reflux for an additional 2 h and concentrated to dryness in vacuo. The resulting solid residue, containing the crude sodium salt of N-substituted methyl oxopiperidine-3-carboxylate (7), was dissolved in anhydrous MeOH (5 mL). To this solution was added N-substituted-4,5-dihydro-1H-imidazol-2-amine (4) (1 mmol), prepared in a separate step as described above. The basic solution was stirred at reflux for 12 h under an argon atmosphere and then cooled in ice water. The cooled reaction mixture was stirred for 1 hour and the precipitated solid was collected by filtration, washed with cold methanol and dried to yield the pure imipridone product of formula (I).

[0086] 4. Synthesis of amine-containing imipridones For the synthesis of target compounds bearing pendant primary amine or cyclic secondary amine moieties, the corresponding mono-Boc-protected diamines (3 or 5) at either the X or Y group were used as coupling components in the general procedure described above. The isolated Boc-protected imipridone (2 mmol) was dissolved in concentrated hydrochloric acid (5 mL), and the resulting solution was heated to reflux for 5 min and cooled to room temperature. The pH of this solution was then set to approximately 13–14 with concentrated aqueous potassium hydroxide. The precipitated amine product was collected by filtration, washed thoroughly with cold water, and dried over potassium hydride pellets in a desiccator.

[0087] 5. Synthesis of azidobenzylimipridone The corresponding aminobenzylimipridone (1 mmol) was dissolved in concentrated hydrochloric acid (5 mL). To this solution, cooled to 0 °C, was added dropwise an aqueous solution of NaNO (137.8 mg, 2 mmol dissolved in 2.5 mL of water). The diazotization reaction was monitored by TLC. After completion, NaN (324 mg, 5 mmol) was added to the reaction mixture at 0 °C, which was then stirred at room temperature for 1 h. The pH of the solution was adjusted to 10–11 by careful addition of solid NaCO. The resulting mixture was extracted with CHCl (2 × 40 mL), and the organic phase was dried over NaSO and then evaporated to dryness. The oily residue was crystallized from n-hexane to give the product as a colorless solid.

[0088] 6. Synthesis of aryl triazolyl benzyl imipridones from azido benzyl imipridones The corresponding azidobenzylimipridone (1 mmol), the terminal alkyne component (1 mmol), and CuI (29.3 mg, 0.15 mmol) were dissolved in DMSO (5 mL). The reaction mixture was stirred at room temperature for 24 h in a sealed vessel and poured into water (50 mL). The precipitated solid was collected by filtration, washed with water (100 mL), and suspended in aqueous ammonia (20 mL). The suspension was stirred for 20 min and filtered. The residue was washed with water (50 mL), dried, and dissolved in a 9:1 mixture of CHCl and MeOH (10 mL). The solution was passed through silica and evaporated. The solid residue was crystallized from ether.

[0089] The present invention will now be described with reference to exemplary embodiments, which should not be construed as limiting the present invention. [Example]

[0090] Example 1: 7-(3,5-Difluorobenzyl)-4-(4-(trifluoromethyl)benzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-301) (Compound I / 30) [ka] Yield: 387 mg (81%). Melting point: 168.0 o C. 1 H-NMR (CDCl3): 7.53 and 7.50 (AA'BB'スピンシステムのA and びB part, J AB =8.9 Hz, 2x2H, H-3",5" and H-2",6", resp.); 6.84 (br dt, J~7 Hz and H-2 Hz, 2H, H-2',6'); 6.67 (tt, J=9.0 Hz and H-2.3 Hz, 1H, H-4'); 5.06 (s, 2H, H-11); 3.88 (s, 4H, H-1 and H-2); 3.60 (s, 2H, H-10); 3.24 (br s, 2H, H-6); 2.64 (t, 2H, J=5.7 Hz, H-8); 2.46 (t, 2H, J=5.7 Hz, H-9). 13 C-NMR (CDCl3): 163.1 (dd, J=250.2 Hz and 15.6 Hz, C-3',5'); 161.3 (C-5); 152.9 (C-3a); 145.8 (C-9a); 142.2 (t, J=8.4 Hz, C-1'); 140.8 (C-1"); 129.6 (qa, J=32.5 Hz, C-4"); 128.8 (C-2",6"); 125.3 (qa, J=3.8 Hz, C-3",5"); 124.4 (qa, J=272.5 Hz, C F3); 111.3 (dd, J=19.3 Hz and 4.9 Hz, C-2',6'); 102.8 (t, J=25.8 Hz, C-4'); 101.6 (C-5a); 61.4 (C-10); 50.6 (C-2); 49.3 (C-6); 48.4 (C-8); 46.9 (C-1); 45.0 (C-11); 26.8 (C-9).

[0091] Example 2: 4-(4-ブロモベンジル)-7-(3,5-ジフルオロベンジル)-2,4,6,7,8,9-ヘキサヒドロイMitro[1,2-a]Milit[3,4-e]Militor-5(1H)-オン(TBP-302)(Compound I / 39)

change

[0092] Example 3: 4-(4-フルオロベンジル)-7-(3,5-ジフルオロベンジル)-2,4,6,7,8,9-ヘキサヒドロイミダゾ[1,2-a]ピリド[3,4-e]ピリミジン-5(1H)-オン(ABB-011)(Compound 1 / 7)

change

[0093] Example 4: 4-(4-クロロベンジル)-7-(3,5-ジフルオロベンジル)-2,4,6,7,8,9-ヘキサヒドロイミダゾ[1,2-a]ピリド[3,4-e]ピリミジン-5(1H)-オン(TBP-333)(Compound I / 124)

change

[0094] Example 5: 4-(3-chlorobenzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-344) (Compound I / 125) [ka] Yield: 302 mg (68%). Melting point: 204.8 o C. 1 H-NMR (CDCl3): 7.40 (br s, 1H, H-2"); 7.30 (m, 1H, H-5"); 7.18-7.15 (m, 2H, H-4",6"); 6.84 (br dt, J~7 Hz and ~2 Hz, 2H, H-2',6'); 6.66 2.63 (t, 2H, J=5.6 Hz, H-8); 2.45 (t, 2H, J=5.6 Hz, H-9). 13 C-NMR (CDCl3): 163.1 (dd, J=248.6 Hz and 14.0 Hz, C-3',5'); 161.3 (C-5); 152.9 (C-3a); 145.7 (C-9a); 142.3 (t, J=8.4 Hz, C-1'); 138.8 (C-1"); 134.1 (C-3"); 128.4 (two combined lines, C-2" and C-4"); 127.6 (C-6"); 126.8 (C-5"); 111.3 (dd, J=19.6 Hz and 5.1 Hz, C-2',6'); 102.7 (t, J=25.3 Hz, C-4'); 101.6 (C-5a); 61.3 (C-10); 50.6 (C-2); 49.4 (C-6); 48.4 (C-8); 46.9 (C-1); 44.9 (C-11); 26.8 (C-9).

[0095] Example 6: rac-4-(4-chlorobenzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-342) (Compound I / 128 (racemic form)) [ka] Yield: 38 mg (8%). Melting point: 86 o C. 1 H-NMR (CDCl3): 7.28 (d, J=8.3 Hz, 2H, H-2",6"); 7.23 (d, J=8.3 Hz, 2H, H-3",5"); 6.84 (br dt, J~7 Hz and ~2 Hz, 2H, H-2',6'); 6.67 (tt, J=9.0 Hz and 2.3 Hz, 1H, H-4'); 6.08 (dd, J=5.1 Hz and 1.6 Hz, 1H, H-11); 4.31 (dd, J=12.8 Hz and 5.1 Hz, 1H, H-12 A ); 4.06 (dd, J=12.8 Hz and 1.6 Hz, 1H, H-12 B ); 3.93-3.85 (m, 3H, H-1 and H-2 A ); 3.78 (m, 1H, H-2 B ); 3.60 (s, 2H, H-10); 3.24 and 3.21 (A and B parts of the AA'BB' spin system, J AB =14.9 Hz, 2x1H, H-6 A and H-6 B ); 2.66 (m, 2H, H-8); 2.50 (m, 2H, H-9). 13C-NMR (CDCl3): 163.1 (dd, J=248.2 Hz and 13.9 Hz, C-3',5'); 161.6 (C-5); 153.5 (C-3a); 146.0 (C-9a); 142.2 (t, J=8.4 Hz, C-1'); 135.2 (C-1"); 133.1 (C-4"); 139.0 (C-2",6"); 128.4 (C-3",5"); 111.4 (dd, J=19.3 Hz and 4.9 Hz, C-2',6'); 102.8 (t, J=25.7 Hz, C-4'); 102.4 (C-5a); 62.9 (C-12); 61.3 (C-10); 57.6 (C-11); 49.5 (C-2); 49.4 (C-6); 48.4 (C-8); 46.5 (C-1); 26.8 (C-9).

[0096] Example 7: R-4-(4-fluorobenzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-339) (Compound I / 129 (R-enantiomer)) [ka] Yield: 51 mg (11%). Melting point: 76 o C. 1 H-NMR (CDCl3): 7.36 (dd, J = 8.8 Hz and 5.8 Hz, 2H, H-2",6"); 6.96 (t, J = 8.3 Hz, 2H, H-3",5"); 6.85 (br dt, J ~ 7 Hz and ~ 2 Hz, 2H, H-2',6'); 6.67 (tt, J = 9.0 Hz and 2.3 Hz, 1H, H-4'); 6.09 (dd, J = 5.3 Hz and 1.6 Hz, 1H, H-11); 4.32 (dd, J = 12.8 Hz and 5.3 Hz, 1H, H-12 A ); 4.07 (dd, J=12.8 Hz and 1.6 Hz, 1H, H-12 B); 3.94-3.85 (m, 3H, H-1 and H-2 A ); 3.81 (m, 1H, H-2 B ); 3.61 (s, 2H, H-10); 3.25 and 3.21 (A and B parts of the AB spin system, J AB =14.9 Hz, 2x1H, H-6 A and H-6 B ); 2.66 (m, 2H, H-8); 2.50 (m, 2H, H-9). 13 C-NMR (CDCl3): 163.1 (dd, J=248.2 Hz and 13.9 Hz, C-3',5'); 162.0 (d, J=245.8 Hz, C-4"); 161.6 (C-5); 153.5 (C-3a); 145.8 (C-9a); 142.1 (t, J=8.4 Hz, C-1'); 132.5 (br s, C-1"); 129.4 (d, J=7.8 Hz, C-2",6"); 115.1 (d, J=21.4 Hz, C-3",5"); 111.3 (dd, J=19.6 Hz and 5.2 Hz, C-2',6'); 102.8 (t, J=25.6 Hz, C-4'); 102.5 (C-5a); 63.1 (C-12); 61.3 (C-10); 57.6 (C-11); 49.5 (two combined lines, C-2 and C-6); 48.3 (C-8); 46.5 (C-1); 26.8 (C-9).

[0097] Example 8: 4-(3-(aminomethyl)benzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-324) (Compound I / 61) [ka] Yield: 376 mg (86%). Melting point: 129.5 o C. 1¹H-NMR (CDCl₃): 7.33 (br s, 1H, H⁻²"); 7.30 (br d, J=7.6 Hz, 1H, H⁻⁶"); 7.22 (t, J=7.6 Hz 1H, H⁻⁵"); 7.15 (br d, J=7.6 Hz, 1H, H⁻⁴"); 6.84 (br dt, J~7 Hz and ~2 Hz, 2H, H⁻²', 6'); 6.66 (tt, J=8.9 Hz and 2.3 Hz, 1H, H⁻⁴'); 5.00 (s, 2H, H⁻¹¹); 3.92-3.83 (m, 4H, H⁻¹ and H⁻²); 3.79 (s, 2H, H⁻¹²); 3.59 (s, 2H, H-10); 3.24 (br s, 2H, H-6); 2.63 (t, 2H, J=5.7 Hz, H-8); 2.45 (t, 2H, J=5.7 Hz, H-9); 1.77 (br s, 2H, N H 2). 13 C-NMR (CDCl3): 163.1 (dd, J=248.4 Hz and 14.1 Hz, C-3',5'); 161.5 (C-5); 153.2 (C-3a); 145.6 (C-9a); 143.2 (C-3"); 142.3 (t, J=8.4 Hz, C-1'); 137.2 (C-1"); 128.6 (C-5"); 127.3 (C-4"); 127.1 (C-2"); 126.2 (C-4"); 111.4 (dd, J=20.0 Hz and 5.0 Hz, C-2',6'); 102.7 (t, J=25.7 Hz, C-4'); 101.7 (C-5a); 61.4 (C-10); 50.7 (C-2); 49.4 (C-6); 48.4 (C-8); 46.9 (C-1); 46.4 (C-12); 45.4 (C-11); 26.7 (C-9).

[0098] Example 9: 4-(3-アミノベンジル)-7-(3,5-ジフルオロベンジル)-2,4,6,7,8,9-ヘキサヒドロイミダゾ[1,2-a]ピリド[3,4-e]ピリミジン-5(1H)-オン(TBP-346)(Compound I / 126)

change

[0099] Example 10: 4-(4-アジドベンジル)-7-(3,5-ジフルオロベンジル)-2,4,6,7,8,9-ヘキサヒドロイミダゾ[1,2-a]ピリド[3,4-e]ピリミジン-5(1H)-オン(CZT-136)

change

[0100] Example 11: 7-(3-Azidobenzyl)-4-(4-(trifluoromethyl)benzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-272) [ka] Yield: 313 mg (65%). Melting point: 128.8 o C. 1 H-NMR (CDCl3): 7.52 and 7.50 (A and B parts of the AA'BB' spin system, J AB =8.9 Hz, 2x2H, H-3",5" and H-2",6", resp.); 7.26 (t, J=7.7 Hz, 1H, H-5'); 7.06 (d, J=7.7 Hz, 1H, H-6'); 6.99 (t, J=1.5 Hz, 1H, H-2'); 6.90 (dd, J=7.7 Hz and 1.5 Hz, 1H, H-4'); 5.06 (s, 2H, H-11); 3.87 (s, 4H, H-1 and H-2); 3.61 (s, 2H,H-10); 3.24 (br s, 2H, H-6); 2.64 (t, 2H, J=5.7 Hz, H-8); 2.45 (t, 2H, J=5.7 Hz, H-9). 13 C-NMR (CDCl3): 161.7 (C-5); 153.0 (C-3a); 145.9 (C-9a); 140.8 (C-1"); 140.2 (C-3'); 140.0 (C-1'); 129.8 (C-5'); 129.6 (qa, J=32.5 128.9 (C-2",6"); 125.5 (C-6'); 125.3 (qa, J=3.5 Hz, C-3",5"); 124.4 (qa, J=272.5 Hz, C 119,4 (C-2'); 118.1 (C4'); 101.7 (C-5a); 61.9 (C-10); 50.6 (C-2); 49.4 (C-6); 48.4 (C-8); 46.9 (C-1); 45.0 (C-11); 26.8 (C-9).

[0101] Example 12: 4-(3-Azidobenzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-347) (Compound I / 127) [ka] Yield: 355 mg (79%). Melting point: 158 o C. 1 H-NMR (CDCl3): 7.23 (t, J=7.9 Hz, 1H, H-5"); 7.18 (br ~d, J~8 Hz 1H, H-6"); 7.10 (br s, 1H, H-2"); 6.87 (br ~d, J~8 Hz 1H, H-4"); 6.84 (br dt, 6.66 (tt, J=8.9 Hz and 2.3 Hz, 1H, H-4'); 5.00 (s, 2H, H-11); 3.90-3.81 (m, 4H, H-1 and H-2); H-10); 3.24 (br s, 2H, H-6); 2.63 (t, 2H, J=5.7 Hz, H-8); 2.45 (t, 2H, J=5.7 Hz, H-9); 13C-NMR (CDCl3): 163.1 (dd, J=248.4 Hz and 13.7 Hz, C-3',5'); 161.3 (C-5); 153.0 (C-3a); 145.6 (C-9a); 142.3 (t, J=9.0 Hz, C-1'); 140.0 (C-3"); 138.9 (C-1"); 129.7 (C-5"); 125.1 (C-6"); 121.2 (C-2"); 118.0 (C-4"); 111.4 (dd, J=20.1 Hz and 5.4 Hz, C-2',6'); 102.7 (t, J=25.7 Hz, C-4'); 101.7 (C-5a); 61.3 (C-10); 50.6 (C-2); 49.4 (C-6); 48.4 (C-8); 46.9 (C-1); 45.0 (C-11); 26.8 (C-9).

[0102] Example 13: 7-benzyl-4-(4-(4-ferrocenyl-1H-1,2,3-triazol-1-yl)benzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (ABB-030) [ka] Yield: 443 mg (71%). Melting point: 165.4 o C. 1 H-NMR (CDCl3): 7.81 (s, 1H, H-12); 7.65 (d, J=7.9 Hz, 2H, H-3",5"); 7.59 (d, J=7.9 Hz, 2H, H-2",6"); 7.31-7.26 (m. 4H, H-2',3',5',6'); 7.22 (m, 1H, H-4'); 5.11 (s, 2H, H-11); 4.74 (t, J=1.8 Hz, 2H, H-15,18); 4.29 (t, J=1.8 Hz, 2H, H-16,17); 4.07 (s, η 5 -C5 H5); 3.87 (br s, 4H, H-1 and H-2); 3.63 (s, 2H,H-10); 3.27 (br s, 2H, H-6); 2.63 (t, 2H, J=5.7 Hz, H-8); 2.44 (t, 2H, J=5.7 Hz, H-9). 13 C-NMR (CDCl3): 161.3 (C-5); 153.0 (C-3a); 147.5 (C-4'); 145.9 (C-9a); 130.1 (C-2",6"); 128.4 (C-2',6'); 127.4 (C-4'); (C-3',5'); 120.1 (C-3",5"); 116.6 (C-12); 102.2 (C-5a); 75.0 (C-14); 69.6 (η 5 - C 68.8 (C-16,17); 66.8 (C-15,18); 62.3 (C-10); 50.6 (C-2); 49.5 (C-6); 48.3 (C-8); 46.8 (C-1); 44.9 (C-11); 26.8 (C-9).

[0103] Example 14: Synthesis of 4-arylmethyl-substituted 7-(3,5-diazidobenzyl)-2,4,6,7,8,9-hexahydro-imidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-ones (compounds I / 138 and I / 139) Sodium azide (0.260 g, 4 mmol), sodium ascorbate (0.119 g, 0.60 mmol), N,N-dimethylethylenediamine (0.080 g, 0.9 mmol), NaOH (0.012 g, 0.30 mmol), the corresponding 7-(3,5-dibromobenzyl)imipridone I / 134 or I / 135 (1 mmol), and CuI (0.057 g, 0.30 mmol) were dissolved in 20 mL of a degassed (argon purging for 30 min) EtOH / HO solvent mixture (7:3) and heated at reflux for 3 h. The reaction mixture was concentrated under reduced pressure to approximately one-third of its original volume and extracted with CHCl (3 × 4 mL). The combined organic phase was washed with water (3 × 50 mL), dried over NaSO, filtered, and concentrated. Final purification is carried out by column chromatography using CH2Cl2 / MeOH (99:1) as eluent.

[0104] Example 15: Synthesis of 4-arylmethyl-substituted 7-(3,5-dicyanobenzyl)-2,4,6,7,8,9-hexahydro-imidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-ones (compounds I / 136 and I / 137) To a stirred solution of the corresponding 7-(3,5-dibromobenzyl)imipridone I / 134 or I / 135 (1 mmol) in DMF (2 mL), zinc cyanide (0.181 g, 1.2 mmol), 1,1'-bis(diphenylphosphino)ferrocene ligand (DPPF, 0.065 g, 0.117 mmol), and finally Pd2dba3 (45.8 mg, 0.05 mmol) were added sequentially. The flask was flushed with nitrogen and stirred in an oil bath at 110-120 °C for 20 h. After cooling to room temperature, the reaction mixture was evaporated under vacuum. The resulting crude product was subjected to flash silica gel column chromatography (eluent, ethyl acetate:hexane (1:4)) to obtain the dicyanobenzyl-substituted imipridone as a pure product.

[0105] Example 16: Synthesis of 4-arylmethyl-substituted 7-(3-thiocyanatobenzyl)-2,4,6,7,8,9-hexahydro-imidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-ones (compounds I / 142 and I / 143) Under an argon atmosphere, a mixture of the corresponding 7-(3-iodobenzyl)imipridone (I / 140 or I / 141) (1.0 mmol), CuSCN (0.12 g, 1.0 mmol), KSCN (0.095 g, 1.0 mmol), and DMF (3 mL) was heated in an oil bath with stirring and maintained at 140 °C for 12 h. After cooling, the mixture was diluted with toluene (5 mL) and water (5 mL) and then filtered through a Celite bed. The aqueous phase was extracted with toluene (2 × 5 mL), and the combined organic phases were washed with water, dried over NaSO, and concentrated. The residue was chromatographed on silica gel (hexane as eluent) to give the thiocyanate product, which was further purified by recrystallization from hexane.

[0106] Example 17: Synthesis of 4-arylmethyl-substituted 7-(3-selenocyanatobenzyl)-2,4,6,7,8,9-hexahydro-imidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-ones (compounds I / 145 and I / 146) To a solution of the corresponding 7-(3-iodobenzyl)imipridones (I / 79 and I / 144) in dioxane (4 mL) was added 6 N HCl (10 mL). The resulting suspension was cooled to 0 °C, and then NaNO (0.415 g, 6 mmol) in water (2 mL) was slowly added. After stirring for 30 min, saturated NaOAc solution (approximately 30 mL) was added portionwise to adjust the pH of the reaction mixture to 5-6. The resulting suspension was poured into a solution of KSeCN (0.793 g, 5.5 mmol) in water (25 mL) at 0 °C. After stirring for 30 min, the reaction mixture was allowed to warm to room temperature and extracted with ether. The organic layer was washed with water and brine, dried over NaSO, and concentrated. The residue was purified by column chromatography on silica using ethyl acetate-hexane (1:5) as the eluent.

[0107] Example 18: In this example, compounds of formula (I) according to the present invention are listed, along with relevant references known from the prior art, and the results of their in vitro antiproliferative assays are presented for representative compounds tested during our systematic experimental work.

[0108] Cell cultures for long-term treatment experiments The tumor growth inhibitory effects of imipridone derivatives were measured using PANC-1 (human pancreatic carcinoma of biliary origin), COLO205 (human colorectal adenocarcinoma), and A2058 (human metastatic melanoma) cells obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK), and EBC-1 (human lung squamous cell carcinoma) cells purchased from the Japanese Research Resources Bank (Tokyo, Japan). PANC-1 cells were maintained in Dulbecco's modified Eagle's medium (DMEM, Lonza, Basel, Switzerland). COLO-205 cell lines were cultured in DMEM medium supplemented with 4500 mg / L d-glucose. EBC-1 cells were cultured in DMEM medium containing 1% non-essential amino acids (NEAA, Gibco® / Invitrogen Corporation, New York, NY, USA) and 1 mM sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA), while the A2058 cell line was grown in RPMI 1640 (Lonza, Basel, Switzerland). For all cell lines, the aforementioned basal medium was supplemented with 10% fetal bovine serum (FBS, Gibco® / Invitrogen Corporation, New York, NY, USA), L-glutamine (2 mmol / L) (Lonza, Basel, Switzerland), and 100 μg / mL penicillin / streptomycin (Gibco® / Invitrogen Corporation, New York, NY, USA). All cell lines were cultured on plastic culture dishes (Sigma-Aldrich, St. Louis, MO, USA or Eppendorf AG, Hamburg, Germany) under standard conditions (37°C, humidified 5% CO atmosphere).

[0109] Viability assay (long-term treatment experiment) Impedance-based assay The cytotoxicity experiments on PANC-1 cells were performed using the impedance-based xCELLigence SP system (ACEA Biosciences, San Diego, CA, USA). A more detailed explanation of the fundamentals of impedance measurement is provided in our previous paper

[22] . Monitoring the change in impedance, which is proportional to the number of cells attached to the electrode surface, provides a highly sensitive method for cytotoxicity studies

[23] . The change in impedance is expressed in the form of a cell index (CI), calculated by the software built into the xCELLigence system (RTCA2.0, ACEA Biosciences, San Diego, CA, USA). IC 50 To determine the concentration that reduces cell viability by 50%, the imipridone to be tested was dissolved in DMSO and further diluted in supplemented DMEM medium to a concentration of 2.5 × 10 -4 ~5×10 -7 A range of concentrations of M was prepared. The steps of our impedance experiments proceeded in the same way as those described in

[24] . Briefly, after obtaining a constant CI value during background measurements, PANC-1 cells (1.5x10 4 Cells / well) were added to the so-called E-plates and their adhesion / spreading was monitored for 24 hours to allow the cell culture to settle into a plateau phase. In a final step, the equilibrated cells were treated with the test compound (final concentration: 2.5 × 10 -5 ~5×10 -8 M), and changes in CI were monitored at 10 kHz for at least 72 hours. For control wells, an appropriate volume ratio of DMSO was added. Three parallel measurements were performed for each measurement. CI values ​​obtained at each concentration 72 hours after treatment were normalized to the CI value of the DMSO control. IC values ​​were calculated for these normalized CI values ​​by fitting the sigmoidal dose-response curve to a nonlinear regression function in OriginPro8 (OriginLab Corporation, Northampton, MA, USA). 50 values ​​were calculated.

[0110] Colorimetric assay The antiproliferative / cytotoxic effects of imipridone on COLO-205 and A2058 cell lines were measured by the alamarBlue assay. Because COLO-205 cells exhibit weak / negligible adhesion and the A2058 cell line fails to establish a stable plateau phase during impedance analysis, this colorimetric assay proved to be a more suitable method for analyzing these cell lines than the xCELLigence system. The cell inoculation and alamarBlue assay procedures were similar to those described in our previous paper

[24] . The main steps were as follows: (i) cells were inoculated into 96-well plates (Sarstedt AG, Nuumbrecht, Germany) at 10 4 cells / well, (ii) test substance was seeded at 2.5 × 10 -5 ~5×10 -8 (iii) treatment with a final concentration of 0.15 mg / mL for 72 hours; (iv) addition of AlamarBlue reagent (0.15 mg / mL, Sigma-Aldrich, St. Louis, MO) dissolved in PBS (phosphate-buffered saline, pH 7.2); and (v) reading of the fluorescence intensity of the sample after 6 to 8 hours of incubation with AlamarBlue reagent. Fluorescence measurements were performed using an LS-50B luminescence spectrometer (Perkin Elmer Ltd., Buckinghamshire, United Kingdom) with the following settings: excitation wavelength = 560 nm, emission wavelength = 590 nm. Each measurement was performed in triplicate. Wells containing an appropriate volume ratio of DMSO were used as controls. The fluorescence intensity of each sample was expressed as a ratio of the fluorescence of the DMSO control. The IC was calculated using the nonlinear regression function of OriginPro8 (OriginLab Corporation, Northampton, MA, USA) to fit a sigmoidal dose-response curve to the normalized fluorescence intensity. 50 values ​​were calculated.

[0111] Statistical evaluation of the resulting data was performed using RTCA2.0 (ACEA Biosciences, San Diego, CA, USA), MS Excel, and OriginPro8 (OriginLab Corporation, Northampton, MA, USA) software. Data obtained from each experiment represent the mathematical mean. IC 50 The standard deviations of the parameters were also obtained using sigmoidal curve fitting.

[0112] Short-term cytotoxicity testing using the MTT assay For short-term cytotoxicity studies, A-431 (human squamous cell carcinoma) cells and U-87 (human primary glioblastoma) cells were cultured in RPMI-1640 medium supplemented with 10% FCS (fetal calf serum, Sigma Ltd.), 2 mM L-glutamine, and 160 mg / mL gentamicin. Cell cultures were maintained in a humidified atmosphere of 5% CO at 37°C. Cells were grown to confluence and plated at 5.0 × 10 cells per well. 3 The cells were distributed into a 96-well plate at an initial cell number of 10. After 24 hours of incubation at 37°C, the cells were treated with compounds in a final volume of 200 μL containing 1.0% v / v DMSO. The cells were then divided into 10 -4 ~10 2Cells were incubated with compounds at concentrations ranging from 0.1 μM for 1 h. Control cells were treated with serum-free medium (RPMI-1640) alone or serum-free medium containing DMSO (c = 1.0 v / v%) for 1 h at 37 °C. After incubation, cells were washed twice with serum-free (RPMI-1640) medium. To measure in vitro cytostatic effects, cells were cultured for an additional 72 h in 10% serum-containing medium. MTT solution (45 mL, 2 mg / mL, final concentration: 0.37 mg / mL) was added to each well. The respiratory chain [26, 27] and other electron transport systems

[28] degrade MTT, thereby forming water-insoluble purple formazan crystals within the cells

[29] . The amount of these crystals can be measured spectrophotometrically and serves as an estimate of the number of mitochondria in the well and, therefore, the number of viable cells

[30] . After 4 h of incubation, cells were centrifuged (900 g) for 5 min, and the supernatant was removed. The obtained formazan crystals were dissolved in DMSO (100 mL), and the optical density (OD) of the samples was measured at λ = 540 and 620 nm, respectively, using an ELISA reader (iEMS Reader, Labsystems, Finland). The OD620 value was subtracted from the OD540 value. The percentage of cytostatic effect was calculated using the following formula: Cytostatic effect (%) = [1 - (OD 処理 / OD) 対照 ] x 100. OD 処理 value and OD 対照 Values ​​correspond to the optical density of treated and control cells, respectively. In each case, two independent experiments were performed with four parallel measurements. The 50% inhibitory concentration (IC) was calculated from the dose-response curves. 50 The curves were defined using Microcal™ Origin1 (version 7.5) software: cell growth inhibition was plotted as a function of concentration and fitted to a sigmoidal curve, and the half maximal inhibitory concentration (IC) was calculated based on this curve. 50 The IC ) value was determined. 50 represents the concentration of compound required for 50% inhibition in vitro.

[0113] High-throughput screening of EBC-1 and H2228 lung cancer cell lines using the CellTiter-Glo luminescent cell viability assay EBC-1 (obtained from JRCB, https: / / cellbank.nibiohn.go.jp / english / ) and H2228 (obtained from ATCC, https: / / www.lgcstandards-atcc.org / ) cell lines were maintained at 37°C in a 5% CO2 humidified incubator according to the instructions provided by JRCB and ATCC. The effect of compounds on cell viability was measured using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI, USA). Cells were seeded at 1000 cells / well in a flat-bottom white 96-well plate (BRANDplates, catalog number: 781965). After 24 hours, the cells were treated with compounds at a concentration of 100 nM for 72 hours. After treatment, the medium was removed and CellTiter-Glo® reagent was added. Untreated cells were used as a control. Luminescence signals were recorded using a microplate reader (BioTek Synergy 2 Multi-Mode Reader, BioTek, Winooski, VT, USA). Cell viability data (% of untreated control cells) were evaluated using Microsoft Excel. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14]

[0114] a Results of high-throughput screening (HTS) on lung cancer cell lines EBC-1 and H2228 are shown. Compounds that proved potent in HTS were evaluated using IC 50 The values ​​were determined (Table 3).

[0115] Example 19: Several representative imipridones and ONC212 as a reference were further tested in the following human malignant cell lines: PC3 and LNCap (prostate cancer); BxPC3, MiaPaCa2, and Panc1 (pancreatic cancer); A549, HCC827, H1993, and H520 (lung cancer); MDA-MB-453 and MDA-MB-231 (breast cancer) (see Table 2 and Figures 1–11).

[0116] Cell lines were maintained at 37°C in a 5% CO2 humidified incubator according to the instructions provided by ATCC (https: / / www.lgcstandards-atcc.org / ). The effect of compounds on cell viability was measured using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI, USA). Cells were seeded at 1000 cells / well in a flat-bottom white 96-well plate (BRAND plate, catalog number: 781965). After 24 hours, cells were treated with 3-fold serially diluted compound concentrations (300 nM, 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, 1.2 nM) for 72 hours. After treatment, the medium was removed and CellTiter-Glo® reagent was added. Luminescence signals were recorded using a microplate reader (BioTek Synergy 2 Multi-Mode Reader, BioTek, Winooski, VT, USA).

[0117] Cell viability data (% of untreated control cells) were evaluated in Microsoft Excel. Dose-response curves (using a nonlinear regression model, log(inhibitor) vs. response, variable slope) were constructed and IC values ​​calculated using Graph Pad Prism 5.02 software (GraphPad Software, San Diego, CA, USA). 50 value was determined.

[0118] In these studies, our compounds exhibited IC in the low nanomolar range, far superior to ONC212. 50 These compounds have proven to be highly efficient antiproliferative agents characterized by their high therapeutic value. In particular, TBP-301 and TBP-302 can be considered as highly potent anticancer agents. Meanwhile, the potent azide derivatives CZT-136 and TBP-272 offer unique possibilities to identify cellular targets and reveal specific mechanisms of action, which are essential for drug approval. [Table 2]

[0119] Example 20: Additional data to demonstrate the efficacy of the compounds of the present invention. [Table 3]

[0120] Example 21: Cell viability assay protocol The effects of selected compounds on cancer cell viability were measured using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's instructions. Cells were seeded into flat-bottom white 96-well plates (BRANDplates®, catalog number: 781965). The seeded cell density was optimized based on cell size and proliferation rate as follows: Panc-1: 750 cells / well; DU145, PC-3, Capan-1, MIAPaCa-2, SCC-25, FaDu, and EBC-1: 1000 cells / well; LNCaP, Detroit 562, MDA-MB-231, and MDA-MB-453: 1500 cells / well. After 48 h of incubation, cells were treated with 3-fold serially diluted compound concentrations (ranging from 300 nM to 1.2 nM) for 72 h. Untreated cells (incubated in the corresponding cell culture medium for 72 hours) served as a control. After treatment, luminescence signals were recorded using a microplate reader (BioTek Synergy 2 Multi-Mode Reader, BioTek, Winooski, VT, USA). Dose-response curves were generated using a nonlinear regression model (variable slope, four parameters) with GraphPad Prism 8.4.2 software (see Figures 12-51). [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] [Table 13] [Table 14] [Table 15]

[0121] Example 22: This example demonstrates the cytotoxic effect of some compounds according to the invention on the Panc-1 human cell line. 50 The values ​​were measured on an xCELLigence SP instrument (see Figures 52-61). The table below shows the IC values ​​obtained after 24, 48, 72, and 96 hours of treatment. 50 Indicates the value. [Table 16]

[0122] Example 23: This example demonstrates the antitumor effects of compounds I / 1 (ONC212), I / 7 (ABB-011), I / 124 (TBP-333), and I / 107 (CZT-136) on subcutaneously growing MDA-MB-231 tumor xenografts in SCID mice (immunsuprimized mice).

[0123] MDA-MB-231 human triple-negative breast cancer xenografts were formed by subcutaneous inoculation of tumor cells into the backs of immunodeficient (SCID) mice. The tested material was injected intraperitoneally every 2–3 days for 3 weeks.

[0124] The experimental results showed that all tested compounds reduced tumor volume, but this effect was significant only in the case of I / 124 (TBP-333).

[0125] Experimental Design: Compounds tested: Reference I / 1(ONC212): 0.022 mg / animal / treatment; dose: 0.88 mg / kg 0.036 mg / animal / treatment; dose: 1.466 mg / kg I / 7(ABB-011): 0.0213 mg / animal / treatment; dose: 0.85 mg / kg 0.0355 mg / animal / treatment; dose: 1.416 mg / kg I / 124(TBP-333): 0.022 mg / animal / treatment; dose: 0.89 mg / kg 0.036 mg / animal / treatment; dose: 1.466 mg / kg I / 107(CZT-136): 0.022 mg / animal / treatment, dose: 0.89 mg / kg 0.036 mg / animal / treatment; dose: 1.466 mg / kg and saline containing 1% DMSO as a control.

[0126] Forty SCID mice were included in the study. Each group of animals was kept in a separate cage throughout the experiment. Each cage had an identification card listing the animal's date of birth, the date of tumor cell injection, and the number and sex of the animal. The injection date for each substance was written on the identification card. Eight animals were used per substance, and they were identified by ear scratching.

[0127] The animals used in these studies were cared for in accordance with the Guiding Principles for the Care and Use of Animals from the Declaration of Helsinki, and the studies were approved by the local ethics committee.

[0128] Animals were housed in sterile cages in an individually ventilated cage (IVC) system with a controlled 12-hour light / 12-hour dark cycle, with the light period occurring between 07:00 and 19:00. Temperature and humidity were recorded daily throughout the experiment. Sterile rodent specific quality control diet (VRF1, Special Diets Services Ltd, Witham, UK) and acidified (pH = 3) sterile distilled water were available ad libitum throughout the study.

[0129] Each batch of diet was accompanied by a certificate of analysis detailing the nutritional composition, and the health of the mice was assessed by animal facility staff.

[0130] MDA-MB-231 human triple-negative breast cancer cells were cultured in RPMI 1640 medium (Sigma Chemical Co., St. Louis, MO) supplemented with 10% fetal bovine serum (Sigma) and 1% penicillin-streptomycin (Sigma) at 37°C in a humidified atmosphere of 5% CO. Cells from monolayer cultures were detached with 0.02% EDTA (Sigma), washed twice with serum-free medium, and 1 cell suspension was implanted at 1.3 × 10 cells into the dorsum of SCID mice. 6 The cells were subcutaneously inoculated at 100 cells / animal. When the tumor volume reached a detectable size (approximately 100–200 mm), 3 , 12 days after tumor cell inoculation), the test products were inoculated intraperitoneally.

[0131] All test compounds were provided as powders, which were dissolved in DMSO. For final treatment concentrations, compounds were diluted 1:100 in saline (1% DMSO).

[0132] All test compounds were scheduled to be administered intraperitoneally in 0.3 ml saline containing 1% DMSO three times a week. However, during the experiment, this schedule was changed: from 07.19, daily intraperitoneal injections of 0.5 ml were administered (Table 17). [Table 17]

[0133] Animal weights and tumor sizes were recorded throughout the experiment. The compounds tested did not cause weight loss (Figure 62).

[0134] Tumor volumes were measured three times a week throughout the experiment. All of the tested compounds reduced tumor growth, but the effect was significant only for TBP-333 (Figure 63).

[0135] The antitumor effects of the compounds were also evaluated by measuring the tumor weight of each group after the experiment was completed (Figures 64 and 65). Based on tumor weight, TBP-333 was found to show the highest significant inhibition, but all the tested compounds reduced tumor weight compared to the control.

[0136] The results showed that the tested drugs had antitumor effects, as measured by both tumor volume and tumor mass at the end of the experiment, but this effect was only significant in the case of TBP-333. It is possible that even greater effects could be achieved by improving the solubility of the materials or somehow increasing the amount delivered.

[0137] Based on the above results, the following points should be noted: Regarding the influence of the substitution pattern of the benzyl groups attached to the 4- and 7-positions of the imipridone skeleton, we clearly established that [7-(3",5"-difluorobenzyl)]-substituted compounds bearing the same substituent at the 4-position (benzyl group on the N-4 atom) exhibit significantly stronger antiproliferative activity than their [7-(3"-fluorobenzyl)]-substituted counterparts (TBP-301 vs. CZT-021, ABB-011 vs. TBP-218, and TBP-333 vs. TBP-353). This is due to the IC 50 This is clearly reflected in the values ​​(Table 16) and the cell viability data demonstrated in Example 21 obtained by in vitro assays performed on human malignant cell lines by different methods. It must be emphasized that all [7-(3'-fluorobenzyl)]-imipridones, including our original azidobenzyl derivatives, are much more active than ONC-212 in each comparative in vitro test of the present invention. The original monoazido derivatives also showed an extremely good activity profile against the tested cell lines. The IC measured on the PANC-1 cell line was 50 As shown by the values ​​(Table 16), the higher cytotoxicity of [7-(3"-azidobenzyl)]imipridones TBP-272 and TBP-400 over the corresponding [7-(3"-fluorobenzyl)]imipridones CZT-021 and TBP-353, respectively, is a notable advantage and novelty in efficacy profiles. This trend was also evident in assays performed with the CellTiter-Glo® Luminescent Cell Viability Assay on a range of cell lines (see Example 21).

[0138] Finally, all measured IC 50 Taking into account the cell viability data, we can state that the imipridone 4-(4-chlorobenzyl)-7-(3,5-difluorobenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one (TBP-333), which we have prepared and tested for the first time in vitro, is the most potent representative of the members of the small molecule anticancer imipridone family physically identified to date.

[0139] Industrial Applicability The compounds of the present invention have anti-cancer activity, making them suitable for use in medicine.

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Claims

1. Compounds of formula (I) 【Chemistry 1】 [where: When Y is phenyl and Z is H, X is 3-fluorophenyl-methyl (compound I / 44), 3-(aminomethyl)phenyl (compound I / 58), 4-(aminomethyl)phenyl (compound I / 62), 3-azetidinyl (compound I / 90), 4-piperidinyl (compound I / 96), 3-azidophenyl (compound I / 102), 4-azidophenyl (compound I / 104), (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl (compound I / 121), or: 【Chemistry 2】 and or When Y is 3-fluorophenyl and Z is H, X is 2-iodoferrocenyl (compound I / 52), 3-(aminomethyl)phenyl (compound I / 60), 4-(aminomethyl)phenyl (compound I / 63), 3-azidophenyl (compound I / 105), or 4-azidophenyl (compound I / 106); or When Y is 3,5-difluorophenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 61), 4-(aminomethyl)phenyl (compound I / 64), 4-azidophenyl (compound I / 107), 3-azidophenyl (compound I / 127), (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl (compound I / 132); or When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 68); or When Y is 3-fluoro-4-(trifluoromethyl)phenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 73); or When Y is 3-fluorophenylmethyl and Z is H, X is 2-methylphenyl (compound I / 45); or When Y is 3-(aminomethyl)phenyl and Z is H, X is 4-iodophenyl (compound I / 59), ferrocenyl (compound I / 67), 4-(trifluoromethyl)phenyl (compound I / 69), or ferrocenylmethyl (compound I / 70); or When Y is ferrocenyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 65); or When Y is ferrocenylmethyl and Z is H, X is 3-(aminomethyl)phenyl (compound I / 66); or When Y is 3-(methoxycarbonylaminomethyl)phenyl and Z is H, X is ferrocenylmethyl (compound I / 71) or ferrocenyl (compound I / 72); or When Y is 4-aminophenyl and Z is H, X is 3-azetidinyl (compound I / 91) or 4-piperidinyl (compound I / 93); or When Y is 3-azetidinyl and Z is H, X is 2-methylphenyl (compound I / 92); or When Y is 4-piperidinyl and Z is H, X is 2-methylphenyl (compound I / 94) or 4-fluorophenyl (compound I / 95); or When Y is 3-pyrrolidinyl and Z is H, X is 2-methylphenyl (compound I / 97) or 4-fluorophenyl (compound I / 98); or When Y is 2-pyrrolidinyl and Z is H, X is 2-methylphenyl (compound I / 99) or 4-fluorophenyl (compound I / 100); or When Y is 4-azidophenyl and Z is H, X is 2-methylphenyl (compound I / 101); 3-fluorophenyl (compound I / 108), 4-fluorophenyl (compound I / 109), 4-(trifluoromethyl)phenyl (compound I / 110), 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 114), 4-iodophenyl (compound I / 115), 3,4,5-trimethoxyphenyl (compound I / 116), 4-azidophenyl (compound I / 117), 3-azetidinyl (compound I / 118), or 4-piperidinyl (compound I / 119); or When Y is 3-azidophenyl and Z is H, X is 2-methylphenyl (compound I / 103), 4-(trifluoromethyl)phenyl (compound I / 111), 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 112), 4-iodophenyl (compound I / 113), or 4-chlorophenyl (compound I / 133); or When Y is (4-ferrocenyl-1H-1,2,3-triazol-1-yl)phenyl and Z is H, X is 2-methylphenyl (compound I / 120); or Y is 【Transformation 3】 where X is 2-methylphenyl (compound I / 122); or When Y is 3,5-diazidophenyl and Z is H, X is 4-chlorophenyl (compound I / 138) or 4-(trifluoromethyl)phenyl (compound I / 139); or When Y is 3-thiocyanatophenyl and Z is H, X is 4-chlorophenyl (compound I / 142) or 4-(trifluoromethyl)phenyl (compound I / 143); or when Y is 3-selenocyanatophenyl and Z is H, X is 4-chlorophenyl (compound I / 145) or 4-(trifluoromethyl)phenyl (compound I / 146); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein When Y is 3-fluorophenyl and Z is H, X is 4-azidophenyl (compound I / 106); or When Y is 3,5-difluorophenyl and Z is H, X is 4-azidophenyl (compound I / 107); or When Y is 3-azidophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 111) or 4-chlorophenyl (compound I / 133); or when Y is 3,5-diazidophenyl and Z is H, X is 4-chlorophenyl (compound I / 138) or 4-(trifluoromethyl)phenyl (compound I / 139); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

3. 2. The compound of claim 1 , When Y is 3,5-difluorophenyl and Z is H, X is 4-azidophenyl (compound I / 107); or When Y is 3-azidophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 111) or 4-chlorophenyl (compound I / 133); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

4. Compounds of formula (I) for use as pharmaceuticals 【Chemistry 4】 [where: When Y is phenyl and Z is H, X is 4-fluorophenyl (compound I / 5), 3-fluorophenyl (compound I / 8), 3,4,5-trifluorophenyl (compound I / 9), 2,3,4-trifluorophenyl (compound I / 29), 2-fluoro-4-nitrophenyl (compound I / 48), or 3-aminophenyl (compound I / 75); or When Y is 3-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 31), 4-iodophenyl (compound I / 46), 2-fluoro-4-nitrophenyl (compound I / 49), 2-methylphenyl (compound I / 53), or 4-aminophenyl (compound I / 84); or When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-iodophenyl (compound I / 38), 4-bromophenyl (compound I / 39), 2-fluoro-4-nitrophenyl (compound I / 50), 4-chlorophenyl (compound I / 124), or 3-aminophenyl (compound I / 126), or 2-methylphenyl (compound I / 37); or When Y is 2-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 26) or 4-iodophenyl (compound I / 54); or When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 27); or When Y is 4-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 28) or 4-iodophenyl (compound I / 55); or When Y is 4-aminophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 81), 4-fluorophenyl (compound I / 86), 4-iodophenyl (compound I / 87), or 3,4,5-trimethoxyphenyl (compound I / 89); or When Y is 3,5-difluorophenyl and Z is hydroxymethyl, X is 4-chlorophenyl (racemic) (compound I / 128), 4-fluorophenyl (R-enantiomer) (compound I / 129(R)), or 4-fluorophenyl (S-enantiomer) (compound I / 129(S)); When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136) or 4-(trifluoromethyl)phenyl (compound I / 137); or When Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

5. A compound of formula (I) for use in the treatment of cancer 【Transformation 5】 [where: When Y is phenyl and Z is H, X is 4-fluorophenyl (compound I / 5), 3-fluorophenyl (compound I / 8), 3,4,5-trifluorophenyl (compound I / 9), 2,3,4-trifluorophenyl (compound I / 29), 2-fluoro-4-nitrophenyl (compound I / 48), or 3-aminophenyl (compound I / 75); or When Y is 3-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 31), 4-iodophenyl (compound I / 46), 2-fluoro-4-nitrophenyl (compound I / 49), 2-methylphenyl (compound I / 53), or 4-aminophenyl (compound I / 84); or When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-iodophenyl (compound I / 38), 4-bromophenyl (compound I / 39), 2-fluoro-4-nitrophenyl (compound I / 50), 4-chlorophenyl (compound I / 124), or 3-aminophenyl (compound I / 126), or 2-methylphenyl (compound I / 37); or When Y is 2-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 26) or 4-iodophenyl (compound I / 54); or When Y is 4-(trifluoromethyl)phenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 27); or When Y is 4-fluorophenyl and Z is H, X is 3-fluoro-4-(trifluoromethyl)phenyl (compound I / 28) or 4-iodophenyl (compound I / 55); or When Y is 4-aminophenyl and Z is H, X is 4-(trifluoromethyl)phenyl (compound I / 81), 4-fluorophenyl (compound I / 86), 4-iodophenyl (compound I / 87), or 3,4,5-trimethoxyphenyl (compound I / 89); or When Y is 3,5-difluorophenyl and Z is hydroxymethyl, X is 4-chlorophenyl (racemic) (compound I / 128), 4-fluorophenyl (R-enantiomer) (compound I / 129(R)), or 4-fluorophenyl (S-enantiomer) (compound I / 129(S)); When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136) or 4-(trifluoromethyl)phenyl (compound I / 137); or When Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11); Or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or a pharmaceutically acceptable salt thereof.

6. 6. The compound for use according to claim 5, wherein the cancer is selected from the group consisting of prostate cancer, pancreatic cancer, lung cancer, breast cancer, glioma, head and neck cancer, colon cancer, and skin cancer.

7. Compounds for use according to claims 4 to 6, wherein When Y is 3-fluorophenyl and Z is H, X is 2-methylphenyl (compound I / 53); or When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), 4-bromophenyl (compound I / 39), 4-chlorophenyl (compound I / 124), or 2-methylphenyl (compound I / 37); or When Y is 3,5-dicyanophenyl and Z is H, X is 4-chlorophenyl (compound I / 136) or 4-(trifluoromethyl)phenyl (compound I / 137); or When Y is phenyl and Z is H, X is 2-iodophenyl (compound I / 11); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers or a pharmaceutically acceptable salt thereof.

8. Compounds for use according to claims 4 to 6, wherein When Y is 3,5-difluorophenyl and Z is H, X is 4-fluorophenyl (compound I / 7), 4-(trifluoromethyl)phenyl (compound I / 30), or 4-chlorophenyl (compound I / 124); or a stereoisomer, enantiomer, mixture of enantiomers, mixture of diastereoisomers, or pharmaceutically acceptable salt thereof.

9. A compound according to claims 1 to 3 for use as a medicament.

10. A compound according to claims 1 to 3 for use in the treatment of cancer.

11. 11. The compound for use according to claim 10, wherein the cancer is selected from the group consisting of prostate cancer, pancreatic cancer, lung cancer, breast cancer, glioma, head and neck cancer, colon cancer, and skin cancer.

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