Anti-cancer compounds

Abstract

The present invention relates to novel agents and their use in the treatment of proliferative diseases, such as cancer, in particular brain cancer.

Description

Field of the Invention

[0001] The present invention relates to pharmaceutical agents and their use in the treatment of proliferative diseases such as cancer, particularly brain cancer. Background of the Invention

[0003] Existing methods for treating solid brain cancer (i.e., brain tumors) involve one or more of surgery, radiotherapy, and chemotherapy. For example, the Stupp regimen is used to treat glioblastoma (which is the most common brain cancer in humans). This involves radiation / temozolomide chemotherapy together, and then adjuvant chemotherapy with temozolomide alone, and is performed after the maximum degree of surgical resection of the tumor. Temozolomide prolongs survival by about 3 months (compared to radiotherapy alone), and the average survival of glioblastoma patients is 15 months. Avastin has been approved for recurrent glioblastoma, but there is little improvement in survival rate.

[0004] Furthermore, even though 50% of glioblastomas are dependent on epidermal growth factor receptor (EGFR) signaling, clinically available EGFR inhibitors have failed in glioblastoma clinical trials. Some inhibitors do not have adequate blood-brain barrier (BBB) ​​permeability. Recent studies have also shown that glioblastomas respond only to type II EGFR inhibitors, while type I inhibitors are being tested. The extreme heterogeneity and aggressiveness of glioblastomas have also led to the failure of molecular targeted therapy as an effective therapy for brain cancer.

[0005] Another class of compounds that have been shown to be effective in many non-brain cancers is tubulin-targeted chemotherapeutics. However, the tubulin inhibitors (e.g., paclitaxel) used clinically are very large molecules that cannot penetrate the blood-brain barrier. In addition, paclitaxel and other tubulin-targeted chemotherapeutics (e.g., vinblastine and vincristine) have serious side effects (e.g., chemotherapy-induced peripheral neuropathy).

[0006] Therefore, there is a need to find new treatments for proliferative diseases such as cancer, and in particular to find effective treatments for brain cancer.

[0007] The reference to any prior art in this specification is not an acknowledgement or implication that the prior art forms part of the common general knowledge in any jurisdiction, or that the prior art could reasonably be expected to be understood by a person skilled in the art to be related and / or combinable with other prior art. SUMMARY OF THE INVENTION

[0008] The present invention seeks to solve one or more of the above problems and / or to provide improvements in cancer treatment and in a first aspect, provides a compound of formula (I):

[0009]

[0010] or a pharmaceutically acceptable salt or prodrug thereof, wherein:

[0011] X is a C1-C6 alkyl group or a C2-C6 alkenyl group;

[0012] Y is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0013] W is O, S or NH;

[0014] Z is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0015] R1 is halogen, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, which cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted;

[0016] Ar is aryl or heteroaryl;

[0017] R2 is H, OH, NH2 or NO2.

[0018] The compound may be a compound of formula (I):

[0019]

[0020] or a pharmaceutically acceptable salt or prodrug thereof, wherein:

[0021] X is a C1-C6 alkyl group or a C2-C6 alkenyl group;

[0022] Y is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0023] W is O, S or NH;

[0024] Z is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0025] R1 is halogen, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, which cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted;

[0026] Ar is aryl or heteroaryl;

[0027] R2 is H, OH, NH2 or NO2.

[0028] Provided that the compound does not have the following structure:

[0029]

[0030] The compound may be a compound of formula (I):

[0031]

[0032] or a pharmaceutically acceptable salt or prodrug thereof, wherein:

[0033] X is a C1-C6 alkyl group or a C2-C6 alkenyl group;

[0034] Y is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0035] W is O, S or NH;

[0036] Z is CH2, NH, N-alkyl, N-alkenyl, S or O;

[0037] R1 is halogen, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, which cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted;

[0038] Ar is aryl or heteroaryl;

[0039] R2 is H, OH, NH2 or NO2.

[0040] Provided that when R1 is aryl (especially phenyl), R1 is not substituted by F.

[0041] X may be a C2 alkyl or C2 alkenyl group.

[0042] Y may be CH2. Z may be NH. Z may be O. Z may be CH2.

[0043] Y may be NH. Y may be O.

[0044] Both Y and Z may be NH.

[0045] Y may be CH2 and Z may be NH.

[0046] Z may be CH2 and Y may be NH.

[0047] Y may be CH2 and Z may be O.

[0048] Y may be O and Z may be CH2.

[0049] W can be O.

[0050] R1 may be a halogen group (e.g., Br). R1 ​​may be an aryl group. The aryl group may be a monocyclic or bicyclic group. The aryl group may be a phenyl group or a naphthyl group.

[0051] The aryl group may be substituted. The substituent may be selected from a halogen group and a heteroalkyl group. The halogen group may be F, and the heteroalkyl group may be an O-alkyl group (e.g., -OCH3) or an aminoalkyl group (e.g., -CH2NH2).

[0052] R1 can be a heteroaryl. The heteroaryl can be a monocyclic or bicyclic ring. The heteroaryl can include one or more nitrogen atoms. For example, the heteroaryl can be pyrazole, isoxazole, triazole, pyridine, pyrimidine, quinoline, benzimidazole or indole. The heteroaryl can be substituted. For example, the substituent can be a halogen group (e.g., F) or a heteroalkyl group (e.g., O-alkyl, such as -OCH3, or aminoalkyl, such as -CH2NH2).

[0053] R1 can be heterocycloalkyl.The heterocycloalkyl can include one or more nitrogen atoms.The heterocycloalkyl can be piperazine.The heterocycloalkyl can include one or more oxygen atoms (except one or more nitrogen atoms, or as a substitute for one or more nitrogen atoms).The heterocycloalkyl can be morpholine.The heterocycloalkyl can be substituted by, for example, a halogen group (for example F) or assorted alkyl (for example O-alkyl, such as-OCH3, or aminoalkyl, such as-CH2NH2).

[0054] Ar can be an aryl group. The aryl group can be a phenyl group. In one embodiment, Ar is a heteroaryl group. The heteroaryl group can include one or more nitrogen atoms and / or NH groups. The heteroaryl group can have 4 or 5 ring carbon atoms. The heteroaryl group can be pyridine or pyrimidine.

[0055] R2 can be H or OH. R2 can be in the para position.

[0056] The compound of formula (I) may be selected from:

[0057]

[0058]

[0059]

[0060] The compound of formula (I) may be selected from:

[0061] 4-(4-Bromophenyl)-N-(4-hydroxyphenyl)butyramide

[0062] 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butanamide

[0063] 4-(4-(2,6-difluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butanamide

[0064] 4-(4-(6-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butyramide

[0065] N-(4-Hydroxyphenyl)-4-(4-(pyridin-3-yl)phenyl)butanamide

[0066] N-(4-Hydroxyphenyl)-4-(4-(2-methoxypyrimidin-5-yl)phenyl)butanamide

[0067] 4-(4-Bromophenyl)-N-phenylbutyramide

[0068] 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-phenylbutyramide

[0069] N-(4-Hydroxyphenyl)-4-(4-(pyrimidin-5-yl)phenyl)butanamide

[0070] 4-(4'-(Aminomethyl)-[1,1'-biphenyl]-4-yl)-N-(4-hydroxyphenyl)butyramide

[0071] N-(4-Hydroxyphenyl)-4-(4-(quinolin-3-yl)phenyl)butanamide.

[0072] In a second aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) (according to the first aspect of the invention) and a pharmaceutically acceptable carrier, diluent or excipient.

[0073] The compounds and pharmaceutical compositions of the present invention may be suitable for the treatment or prevention of proliferative diseases. Therefore, in another aspect, the present invention relates to a method for treating or preventing a proliferative disease in a subject, the method comprising administering to the subject an effective amount of a compound of formula (I) of the first aspect of the present invention or a pharmaceutical composition of the second aspect of the present invention.

[0074] In another aspect, the present invention relates to use of the compound of formula (I) according to the first aspect of the present invention or the pharmaceutical composition according to the second aspect of the present invention in the preparation of a medicament for treating or preventing a proliferative disease.

[0075] In another aspect, the present invention relates to the use of the compound of formula (I) of the first aspect of the present invention or the pharmaceutical composition of the second aspect of the present invention in treating or preventing a proliferative disease in a subject.

[0076] In another aspect, the present invention relates to a compound of formula (I) according to the first aspect of the present invention or a pharmaceutical composition according to the second aspect of the present invention for use in treating or preventing a proliferative disease in a subject.

[0077] In one embodiment, the proliferative disease is cancer. The cancer may be selected from the group consisting of brain cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, uterine cancer, skin cancer, colon cancer, and bladder cancer.

[0078] The cancer may be primary. The cancer may be metastatic. The cancer may be benign. The cancer may be malignant.

[0079] The cancer can be a brain cancer (e.g., anaplastic astrocytoma, astrocytoma, central neurocytoma, choroid plexus carcinoma, choroid plexus papilloma, choroid plexus tumor, diffuse intrapontine glioma, dysembryoplastic neuroepithelial tumor, ependymal tumor, fibrillary astrocytoma, giant cell glioblastoma, glioblastoma multiforme, glioma cerebri, gliosarcoma, hemangiopericytoma, medulloblastoma, medulloepithelioma, meningeal carcinoma, neuroblastoma, neurocytoma, oligoastrocytoma, oligodendroglioma, optic nerve sheath brain tumor, pediatric ependymoma, fibrillary astrocytoma, pinealoblastoma, pinealoma, polymorphic anaplastic neuroblastoma, polymorphic xanthoastrocytoma, primary central nervous system lymphoma, sphenoid ridge meningioma, subependymoma giant cell astrocytoma, subependymoma, trilateral retinoblastoma). The brain cancer may be a primary cancer (eg, a glioma, a meningioma, a pituitary adenoma, or a schwannoma). The brain cancer may be a metastatic cancer (eg, a result of a melanoma or lung cancer).

[0080] In another aspect, the present invention relates to a method for completely or partially preventing the recurrence of a solid tumor in a subject, the method comprising administering to the subject an effective amount of a compound of formula (I) of the first aspect of the present invention or a pharmaceutical composition of the second aspect of the present invention.

[0081] In another aspect, the present invention relates to use of the compound of the first aspect of the present invention or the pharmaceutical composition of the second aspect of the present invention in the preparation of a medicament for completely or partially preventing the recurrence of solid tumors.

[0082] In another aspect, the present invention relates to use of the compound of formula (I) of the first aspect of the present invention or the pharmaceutical composition of the second aspect of the present invention for completely or partially preventing the recurrence of a solid tumor in a subject.

[0083] In another aspect, the present invention relates to a compound of formula (I) according to the first aspect of the present invention or a pharmaceutical composition according to the second aspect of the present invention for use in completely or partially preventing the recurrence of a solid tumor in a subject.

[0084] The solid tumor may be a brain cancer (e.g., a glioblastoma, an astrocytoma, or a glioma). The brain cancer may be a primary cancer. The brain cancer may be a metastatic cancer.

[0085] The compounds of formula (I) may be used alone in therapy or in combination with one or more other therapeutic agents, for example as part of a combination therapy.

[0086] Further aspects of the invention described in the preceding paragraphs and embodiments thereof will become apparent from the following description given by way of example with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0087] Figure 1 Results of treating various cancer cell lines with SC-38 to test the cancer cell viability and cytotoxicity of SC-38.

[0088] Figure 2 Results of treating various cancer cell lines with SC-38 to test the cytotoxicity of SC-38 to malignant cells.

[0089] Figure 3 . Brain uptake of compounds 1-18.

[0090] Figure 4 SC-38-induced apoptosis in U87 and U87vIII cells.

[0091] Figure 5 . Western blot analysis of PARP cleavage in U87 and U87vIII cells after SC-38 treatment. (A) U87 (lanes 1-3) and U87vIII (lanes 4-6) cells were treated with 1 and 5 μM SC-38 for 48 hours. Representative blots of 3 independent experiments are shown. GAPDH was used as a loading control. (B) PARP cleavage was quantified and normalized to GAPDH (expressed as fold change). Results are mean ± SEM from 3 independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnet post-test (*P<0.05, ***P<0.001, ****P<0.0001).

[0092] Figure 6 . Apoptosis induced by compound 1-08 in U87 and U87vIII cells. U87 (AB) and U87vIII (CD) cells were treated with compound 1-08 for 48 hours and stained with Annexin-FITC / 7-AAD. Apoptotic cells were visualized using MUSE cell analyzer (Millipore). Results are mean ± SEM of 3 independent experiments. Statistical significance was calculated using one-way ANOVA followed by Dunnet post-test (***P<0.001).

[0093] Figure 7. Western blot analysis of PARP cleavage in U87 and U87vIII cells after compound 1-08 treatment. (A) U87 (lanes 1-3) and U87vIII (lanes 4-6) cells were treated with 1 and 5 μM compound 1-08 for 48 hours. Representative blots of 3 independent experiments are shown. GAPDH was used as a loading control. (B) PARP cleavage was quantified and normalized to GAPDH (expressed as fold change). Results are mean ± SEM from 3 independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnet post-test (*P<0.05, ****P<0.0001).

[0094] Figure 8 Effects of SC-38 treatment on Bcl-2 family proteins in U87, U87vIII, and U251.

[0095] Fig. 9 Effect of Compound 1-08 treatment on Bcl-2 family proteins in U87, U87vIII and U251. U87 (lanes 1-3), U87vIII (lanes 4-6) and U251 (lanes 7-9) cells. Representative blots are shown in (A) and quantification is shown in (BE), showing relative protein expression. Results are mean ± SEM from 2 independent experiments, and statistical significance was determined using unpaired t-test (*P<0.05, **P<0.01).

[0096] Fig.10 Compound 1-08 (referred to as "Compound 13" and "Compound 13") exhibits a mechanism of action consistent with microtubule disruption. (A) Compound 1-08 inhibits tubulin polymerization in vitro. Data are presented as the mean from at least 3 independent experiments; each data point was performed three times. (B) U87 cells treated with compound 1-08 (24 hours) were stained with Alexa488-labeled anti-β-tubulin antibody (green) or DAPI (blue). Treatment profoundly disrupted the microtubule network and led to the formation of multinucleated cells (white arrows). The white scale bar represents 50 μm. Multinucleated cells were counted manually, and at least 200 cells were counted for each sample. The data in the bar graph (C) are presented as mean ± SEM (n = 3; *P < 0.05, ***P < 0.001; one-way ANOVA followed by Newman-Keuls post-test). (D) U87 cells were treated with compound 1-08 (48 hours), and tubulin and DNA were stained as in (B). Treatment with 1-08 results in defective mitotic spindle formation. Representative images from 3 independent experiments are shown. White scale bar represents 2 μm.

[0097] Fig.11Efficacy of SC-38 (referred to as "CMPD1"), compound 1-08 (referred to as "Compound 13"), compound 1-18 (referred to as "Compound 7"), and compound 1-38 (referred to as "Compound 10") in primary glioma spheres representing two glioblastoma subtypes. (A) RN1, classical and (B) WK1, mesenchymal. The cellular efficacy of the test compounds was determined using the Alamar Blue cell viability assay after 72 hours of drug treatment (EC 50 ). Data are mean ± SEM from 3 independent experiments performed in triplicate. DETAILED DESCRIPTION

[0098] The present invention is based on the surprising discovery that compounds of formula (I) have unexpectedly improved properties in the treatment of proliferative diseases such as cancer, in particular brain cancer.

[0099] Standard nomenclature is generally used herein to describe compounds. For compounds with asymmetric centers, it is understood that all optical isomers and mixtures thereof are included unless otherwise stated. Compounds with two or more asymmetric elements may also exist as mixtures of diastereomers. In addition, compounds with carbon-carbon double bonds may exist in Z and E forms, and all isomeric forms of the compounds are included in the present invention unless otherwise stated. When compounds exist in various tautomeric forms, the compounds listed are not limited to any one specific tautomer, but are intended to include all tautomeric forms. The compounds listed are further intended to include compounds in which one or more atoms are replaced by isotopes (i.e., atoms with the same atomic number but different mass numbers). As a general example, but not limited to this, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 11 C. 13 C and 14 C.

[0100] The compound according to the chemical formula provided herein with one or more stereoisomeric centers has an enantiomeric excess of at least 50%. For example, such a compound may have an enantiomeric excess of at least 60%, 70%, 80%, 85%, 90%, 95% or 98%. Some embodiments of the compound have an enantiomeric excess of at least 99%. It is apparent that a single enantiomer (optically active form) can be obtained by asymmetric synthesis, synthesis derived from optically pure precursors, biosynthesis or by the resolution of racemates (e.g., enzyme resolution or by conventional methods (e.g., crystallization in the presence of a resolving agent, or chromatography, using, for example, chiral HPLC columns)).

[0101] Certain compounds are described herein using general formulas including variables such as R1, R2, Ar, W, X, Y, and Z. Unless otherwise indicated, the definition of each variable in the formula is independent of any other variable, and any variable that appears more than once in a formula is independently defined at each occurrence. Thus, for example, if a group is substituted with 0, 1, or 2 R*, the group may be unsubstituted or substituted with up to two R* groups, and each occurrence of R* is independently selected from the definition of R*. In addition, combinations of substituents and / or variables are permitted only when such combinations produce stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity).

[0102] The "pharmaceutically acceptable salts" of the compounds disclosed herein are acid or base salts generally considered in the art to be suitable for contact with human or animal tissues without excessive toxicity or carcinogenicity, preferably without irritation, allergic reactions or other problems or complications. In particular, the pharmaceutically acceptable salts of the present invention are those that do not adversely affect the ability of the compound to cross the BBB. Such salts include mineral and organic acid salts of basic residues (such as amines), and alkali or organic salts of acidic residues (such as carboxylic acids).

[0103] Suitable pharmaceutically acceptable salts include, but are not limited to, for example, hydrochloric acid, phosphoric acid, hydrobromic acid, malic acid, glycolic acid, fumaric acid, sulfuric acid, sulfamic acid, para-aminobenzenesulfonic acid, formic acid, toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, ethanedisulfonic acid, 2-hydroxyethylsulfonic acid, nitric acid, benzoic acid, 2-acetoxybenzoic acid, citric acid, tartaric acid, lactic acid, stearic acid, salicylic acid, glutamic acid, ascorbic acid, pamoic acid, succinic acid, fumaric acid, maleic acid, propionic acid, hydroxymaleic acid, hydroiodic acid, phenylacetic acid, alkanoic acids (such as acetic acid, HOOC-(CH2) n -COOH (wherein n is any integer from 0 to 6, i.e., 0, 1, 2, 3, 4, 5 or 6). Similarly, pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium and ammonium. Those skilled in the art will recognize other pharmaceutically acceptable salts of the compounds provided herein. Generally, pharmaceutically acceptable acid or base salts can be synthesized from a parent compound containing a basic or acidic portion by any conventional chemical method. In short, these salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent (e.g., ether, ethyl acetate, ethanol, isopropanol or acetonitrile) or a mixture of the two.

[0104] It is obvious that each compound of formula (I) may, but need not, exist in the form of a hydrate, solvate or non-covalent complex. In addition, various crystalline forms and polymorphs and prodrugs of the compounds of formula (I) provided herein are also within the scope of the present invention.

[0105] "Prodrug" is a compound that may not fully meet the structural requirements of the compounds provided herein, but is modified in vivo after administration to a subject or patient to produce a compound of formula (I) provided herein. For example, a prodrug may be an acylated derivative of a compound provided herein. Prodrugs include compounds in which a hydroxyl, carboxyl, amine, or sulfhydryl group is bonded to any group (when administered to a mammalian subject, these groups are cleaved to form free hydroxyl, carboxyl, amino, or sulfhydryl groups, respectively). Examples of prodrugs include, but are not limited to, acetate, formate, phosphate, and benzoate derivatives of alcohol and amine functional groups in the compounds provided herein. Prodrugs of the compounds provided herein can be prepared by modifying the functional groups in the compound in such a manner that the modification is cleaved in vivo to produce the parent compound.

[0106] As used herein, "substituent" refers to a molecular part that is covalently bonded to an atom in a target molecule. For example, a "ring substituent" can be a part of a halogen, alkyl, heteroalkyl, haloalkyl or other substituent described herein, which is covalently bonded to an atom (preferably a carbon atom or a nitrogen atom) that is a ring member. As used herein, the term "substituted" refers to any one or more hydrogens on a designated atom being replaced by a substituent selected from the indicated substituent, provided that the normal valence of the designated atom is not exceeded, and the substitution forms a stable compound, i.e., a compound that can be isolated, characterized and tested for biological activity. When the substituent is an oxo, i.e., =O, two hydrogens on the atom are replaced. Oxo as a substituent of an aromatic carbon atom causes -CH- to be converted to -C(=O)- and loses aromaticity. For example, an oxo-substituted pyridinyl is a pyridone. Examples of suitable substituents are alkyl (including haloalkyl, e.g. CF3), heteroalkyl (e.g. -OCH3, -CH2NHCH3, -CH2NH2), halogen (e.g. fluorine, chlorine, bromine or iodine atoms), OH, =O, SH, SO3H, NH2, =NH, N3 and NO2 groups.

[0107] The term "alkyl" refers to a saturated, linear or branched hydrocarbon group. Specific examples of the alkyl group are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl and 2,2-dimethylbutyl.

[0108] The term "heteroalkyl" refers to an alkyl group as defined above containing one or more heteroatoms selected from oxygen, nitrogen and sulfur, in particular oxygen and nitrogen. Specific examples of heteroalkyl groups are O-alkyl (e.g., methoxy, trifluoromethoxy, ethoxy, n-propoxy, isopropoxy, butoxy and tert-butoxy), methoxymethyl, ethoxymethyl, —CH2CH2OH, —CH2OH, methoxyethyl, 1-methoxyethyl, 1-ethoxyethyl, 2-methoxyethyl or 2-ethoxyethyl, aminoalkyl (e.g., —CH2NH2, —CH2CH2NH2, etc.), methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, isopropylethylamino, methylaminomethyl, ethylaminomethyl, diisopropylaminoethyl, methylthio, ethylthio, isopropylthio, enol ether, dimethylaminomethyl, dimethylaminoethyl, acetyl, propionyl, butyryloxy, acetoxy, methoxycarbonyl, ethoxycarbonyl, propionyloxy, acetylamino, propionylamino, carboxymethyl, carboxyethyl, carboxypropyl, N-ethyl-N-methylcarbamoyl and N-methylcarbamoyl. Other examples of heteroalkyl groups are nitrile, isonitrile, cyanate, thiocyanate, isocyanate, isothiocyanate and alkylnitrile groups.

[0109] The term "alkenyl" refers to an at least partially unsaturated straight or branched hydrocarbon group containing at least two carbon atoms (i.e., C2 alkenyl). Specific examples of alkenyl are ethenyl, propenyl (allyl), isopropenyl, butenyl, ethynyl, propynyl, butynyl, ethynyl, propargyl, isopentadienyl and hex-2-enyl. Preferably, the alkenyl has one or two double bonds.

[0110] The term "cycloalkyl" refers to a saturated or partially unsaturated (e.g., cycloalkenyl) cyclic radical containing one or more rings (preferably 1 or 2) and containing 3 to 14 ring carbon atoms, preferably 3 to 10 (especially 3, 4, 5, 6 or 7) ring carbon atoms. Specific examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, spiro[4,5]decyl, norbornyl, cyclohexyl, cyclopentenyl, cyclohexadienyl, decahydronaphthyl, bicyclo[4.3.0]nonyl, tetrahydronaphthyl, adamantane (i.e., tricyclo[3.3.1.13,7]decane), cyclopentylcyclohexyl and cyclohex-2-enyl.

[0111] The term "heterocycloalkyl" refers to a cycloalkyl group as defined above, wherein one or more (preferably 1, 2 or 3) ring carbon atoms are each independently replaced by oxygen, nitrogen, silicon, selenium, phosphorus or sulfur atoms (preferably by oxygen, sulfur or nitrogen atoms). This includes groups containing such atoms as NH. Heterocycloalkyl preferably has 1 or 2 rings containing 3 to 10 (particularly 3, 4, 5, 6 or 7) ring atoms (preferably selected from C, O, N and S). Specific examples are piperidinyl, prolinyl, imidazolidinyl, piperazinyl, morpholinyl, hexamethylenetetramine (urotropinyl), pyrrolidinyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrofuranyl and 2-pyrazolinyl as well as lactams, lactones, cyclic imides and cyclic anhydrides.

[0112] The term "aryl" refers to an aromatic group containing one or more rings containing 6 to 14 ring carbon atoms, preferably 6 to 10 (especially 6) ring carbon atoms. Examples are phenyl, naphthyl and biphenyl.

[0113] The term "heteroaryl" refers to an aromatic group containing one or more rings containing 5 to 14 ring atoms, preferably 5 to 10 (especially 5 or 6) ring atoms, and containing one or more (preferably 1, 2, 3 or 4) oxygen, nitrogen, phosphorus or sulfur ring atoms (preferably O, S or N). This includes O, S or N-containing groups, such as NH. Examples are pyridyl (e.g. 4-pyridyl), imidazolyl (e.g. 2-imidazolyl), phenylpyrrolyl (e.g. 3-phenylpyrrolyl), thiazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, oxadiazolyl, thiadiazolyl, indolyl, indazolyl, tetrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, isoxazolyl, indazolyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, pyridazinyl, quinolyl, isoquinolyl, pyrrolyl, purinyl, carbazolyl, acridinyl, pyrimidinyl, 2,3′-difuranyl and pyrazolyl (e.g. 3-pyrazolyl).

[0114] The term "halogen" or "halogen atom" as used herein refers to fluorine, chlorine, bromine or iodine.

[0115] The term "optionally substituted" refers to a group in which one, two, three or more hydrogen atoms are independently substituted by halogen (e.g., fluorine, chlorine, bromine or iodine atoms) and / or by, for example, OH, =O, SH, SO3H, NH2, N-alkyl, NH-alkyl, N3 or NO2 groups. The expression also refers to a group substituted by one, two, three or more alkyl, alkenyl or heteroalkyl (e.g., -OCH3, -OCH2CH3, -CH2NHCH3 and -CH2NH2). These groups themselves can be substituted. For example, an alkyl substituent can be substituted by one or more halogen atoms (i.e., can be a haloalkyl). The term "haloalkyl" refers to an alkyl (as defined above) substituted by one or more halogen atoms (as defined above). The specific examples of haloalkyl are trifluoromethyl, dichloroethyl, dichloromethyl and iodoethyl.

[0116] As used herein, words defining the limits of a length range, such as, for example, "from 1 to 5" refer to any integer from 1 to 5, i.e., 1, 2, 3, 4, and 5. In other words, any range defined by explicitly mentioning two integers is meant to include and disclose any integers defining the limits and any integers contained within the range.

[0117] Preferred compounds of formula (I) are those wherein X is C1, C2 or C3 alkyl or C2 or C3 alkenyl (eg C2 alkyl or C2 alkenyl). X may also be C1 alkyl.

[0118] Y may be CH2. Z may be NH. Z may be O. Z may be CH2.

[0119] Y may be NH. Y may be O.

[0120] Both Y and Z may be NH.

[0121] Y may be CH2 and Z may be NH.

[0122] Z may be CH2 and Y may be NH.

[0123] Y may be CH2 and Z may be O.

[0124] Y may be O and Z may be CH2.

[0125] R1 may be a halogen group (e.g., Br). R1 ​​may be an aryl group. The aryl group may be a monocyclic or bicyclic group. The aryl group may be a phenyl group or a naphthyl group.

[0126] The aryl group may be substituted. The substituent may be selected from a halogen group and a heteroalkyl group. The halogen group may be F, and the heteroalkyl group may be an O-alkyl group (e.g., OCH3 or OCH2CH3) or an aminoalkyl group (e.g., -CH2NH2 or -CH2CH2NH2).

[0127] R1 can be a heteroaryl. The heteroaryl can be a monocyclic or bicyclic ring. The heteroaryl can include one or more nitrogen atoms. For example, the heteroaryl can be pyrazole, isoxazole, triazole, pyridine, pyrimidine, quinoline, benzimidazole or indole. The heteroaryl can be substituted. For example, the substituent can be a halogen group (e.g., F) or a heteroalkyl group (e.g., O-alkyl, such as -OCH3 or OCH2CH3, or aminoalkyl, such as -CH2NH2 or -CH2CH2NH2).

[0128] R1 can be a heterocycloalkyl. The heterocycloalkyl can include one or more nitrogen atoms. The heterocycloalkyl can be piperazine. The heterocycloalkyl can include one or more oxygen atoms. The heterocycloalkyl can be morpholine. The heterocycloalkyl can be substituted by, for example, a halogen group (e.g., F) or a heteroalkyl (e.g., O-alkyl, such as -OCH3 or OCH2CH3, or an aminoalkyl, such as -CH2NH2 or -CH2CH2NH2).

[0129] Ar may be an aryl group. The aryl group may be a phenyl group. Ar may be a heteroaryl group. The heteroaryl group may include one or more nitrogen atoms. The heteroaryl group may have 4 or 5 ring carbon atoms. The heteroaryl group may be pyridine or pyrimidine.

[0130] R2 can be H or OH. R2 can be in the para position.

[0131] Specific examples of the compounds of the present invention are listed in Table 1 below.

[0132] Table 1. Examples of compounds of the present invention

[0133]

[0134]

[0135]

[0136] In one embodiment, the compound of formula (I) is selected from compounds 1-01, 1-07, 1-08, 1-17, 1-18, 1-19, 1-20, 1-21, 1-22, 1-38 and 20 from Table 1 above.

[0137] In another embodiment, the compound of formula (I) does not include compound SC-38 from Table 1 above.

[0138] In one embodiment, the compound of the invention is a compound of formula (I) as described herein, with the proviso that the compound does not have the following structure:

[0139]

[0140] In one embodiment, the compound of the invention is a compound of formula (I) as described herein, with the proviso that when R1 is aryl (particularly phenyl), R1 is not substituted by F.

[0141] The compounds of the present invention can be synthesized by any suitable method known to those skilled in the art. A general synthesis is given below in Scheme 1.

[0142]

[0143] Scheme 1. Examples of general syntheses of compounds of the invention

[0144] The compounds of the present invention may exhibit high antiproliferative activity, particularly high efficacy against brain cancer. Specifically, in the examples herein, specific compounds were shown to induce apoptosis and also be able to cross the BBB.

[0145] The therapeutic use of the compound of formula (I), its pharmaceutically acceptable salt, solvate, hydrate, prodrug and preparation and pharmaceutical composition (including mixture of the compound of formula (I)) is within the scope of the present invention. Therefore, the present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I) or its pharmaceutically acceptable salt, solvate, hydrate or prodrug and one or more pharmaceutically acceptable excipients.

[0146] "Pharmaceutical carriers, diluents or excipients" include, but are not limited to, any physiologically buffered (i.e., pH about 7.0 to 7.4) medium, including suitable water-soluble carriers, conventional solvents, dispersion media, fillers, solid carriers, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents. Suitable water-soluble carriers include, but are not limited to, saline, glucose, corn oil, dimethyl sulfoxide, and gelatin capsules. Other conventional additives include lactose, mannitol, corn starch, potato starch, binders (e.g., crystalline cellulose, cellulose derivatives, gum arabic, gelatin), disintegrants (e.g., sodium carboxymethyl cellulose), and lubricants (e.g., talc or magnesium stearate).

[0147] The pharmaceutical composition can be formulated for any suitable route of administration, including, for example, topical (e.g., transdermal or ocular), oral, buccal, nasal, vaginal, rectal, or parenteral administration. The term "parenteral" as used herein includes subcutaneous, intradermal, intravascular (e.g., intravenous), intramuscular, spinal, intracranial, intrathecal, intraocular, periocular, intraorbital, intrasynovial, and intraperitoneal injections and any similar injection or infusion techniques. In some embodiments, compositions suitable for oral use or parenteral use are preferred. Suitable oral forms include, for example, tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. For intravenous, intramuscular, subcutaneous, or intraperitoneal administration, one or more compounds can be combined with a sterile aqueous solution, which is preferably isotonic with the recipient blood. Such preparations can be prepared by dissolving the solid active ingredient in water containing a physiologically compatible substance (such as sodium chloride or glycine) and making it have a buffered pH compatible with physiological conditions to produce an aqueous solution, and making the solution aseptic. The preparation can be present in a single or multi-dose container, such as a sealed ampoule or vial. Examples of suitable components are described in Martindale-The Extra Pharmacopoeia (Pharmaceutical Press, London, 1993) and Martin (ed.), Remington's Pharmaceutical Sciences.

[0148] For the treatment of proliferative diseases, the dosage of the bioactive compounds of the present invention can vary over a wide range and can be adjusted according to individual needs. The active compound of the present invention is usually applied in a therapeutically effective amount. The preferred dosage range is about 0.1 mg to about 140 mg per kilogram of body weight per day (for example, about 0.5 mg to about 7 g per patient per day). The daily dose can be applied in a single dose or in multiple doses. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary according to the treated host and a specific mode of administration. The dosage unit form usually contains about 1 mg to about 500 mg of active ingredient.

[0149] However, it should be understood that the specific dosage level for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, route of administration and excretion rate, drug combination (i.e., other drugs used to treat the patient), the severity of the specific condition being treated, and the location of the unwanted proliferative cells. If the compound is applied locally rather than systemically, and is used for prevention rather than treatment, the dosage is generally lower. Such treatment can be administered as much as possible as needed and administered within the necessary time period as determined by the treating physician. It will be understood by those skilled in the art that it may be necessary to optimize the dosage regimen or therapeutically effective amount of the compound of formula (I) to be administered for each individual.

[0150] It should be understood that different doses may be required to treat different diseases. An effective amount of a drug is an amount that results in a statistically significant decrease in tumor cell count, growth or size. Neoplastic diseases that respond to the drugs of the present invention include, but are not limited to, brain cancer.

[0151] The term "therapeutically effective amount" or "effective amount" refers to the amount of the compound of formula (I) that results in prevention, improvement or repair of proliferative disorder symptoms. The dosage form and dose of the compound or pharmaceutical composition of the present invention can be easily determined by reference to known treatment or prevention regimens.

[0152] Preferred compounds of the present invention will possess certain pharmacological properties, including but not limited to oral bioavailability and BBB permeability, such that the preferred oral dosage forms discussed above can provide therapeutically effective levels of the compound in vivo.

[0153] The compounds of the invention are preferably administered orally or parenterally to a patient (eg, a human) and are present in at least one body fluid or tissue of the patient. Thus, the invention also provides methods for treating a patient suffering from a proliferative disease (including cancer, such as brain cancer).

[0154] The terms "treat", "treatment" and "therapy" are used herein to refer to therapeutic treatments. Therefore, in the present context, the term "treat" includes both curing and ameliorating the severity of cancer or its associated symptoms.

[0155] If cancer develops after administration of the compounds or pharmaceutical compositions of the present invention, "preventing" or "prevention" refers to preventing the occurrence of cancer or ameliorating the severity of cancer. This may be preventing the occurrence of clinically apparent undesirable cell proliferation altogether or preventing the occurrence of preclinically apparent stages of undesirable rapid cell proliferation in individuals at risk.

[0156] Patients may include, but are not limited to, primates, particularly humans, domesticated companion animals (eg, dogs, cats, horses), and livestock (eg, cattle, pigs, sheep) with dosages as described herein.

[0157] The compounds of the present invention can be used to treat and / or prevent disorders and disorders (including cancer, such as brain cancer) associated with cell proliferation. Therefore, the present invention also relates to a method for treating or preventing a proliferative disease in a patient, comprising administering to the patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. The present invention also relates to the use of a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof for treating or preventing proliferative disorders. In any embodiment described in this specification, the present invention also provides a pharmaceutical composition for treating or preventing proliferative disorders. The present invention also relates to the use of a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof in the preparation of a medicament for treating or preventing proliferative disorders.

[0158] When used in a method for treating or preventing a proliferative disorder, the present invention further relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. The present invention further relates to a composition having an active ingredient for treating or preventing a proliferative disorder, wherein the active ingredient is a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. The present invention further relates to the use of a pharmaceutical composition containing a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof in treating or preventing a proliferative disorder (as described above). In one embodiment, the compound of formula (I) is substantially the only active ingredient of the composition. In one embodiment, the proliferative disorder is cancer. In one embodiment, the cancer is brain cancer (e.g., a solid tumor).

[0159] The compounds of formula (I) of the present invention and their pharmaceutical compositions can be used to treat or prevent proliferative diseases, preferably cancer. The compounds and compositions of the present invention can be used to treat a variety of cancers (tumors), including but not limited to: solid tumors, such as brain cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, uterine cancer, brain cancer, skin cancer, colon cancer and bladder cancer.

[0160] The types of cancer or tumor cells that the present invention may be useful for treating include, for example, breast cancer, colon cancer, lung cancer and prostate cancer, gastrointestinal cancer (including esophageal cancer, gastric cancer, colorectal cancer, polyps associated with colorectal tumors, pancreatic cancer and gallbladder cancer), adrenocortical carcinoma, ACTH-producing tumors, bladder cancer, brain cancer (including those discussed below), Ewing's sarcoma, head and neck cancer including oral cancer and laryngeal cancer, kidney cancer including renal cell carcinoma, liver cancer, lung cancer including small cell and non-small cell lung cancer, malignant peritoneal effusion, malignant pleural effusion, skin cancer including malignant melanoma, tumor progression of human skin keratinocytes, squamous cell carcinoma, basal cell carcinoma and hemangiopericytoma, mesothelioma, Kaposi's sarcoma (Kaposi's In one embodiment, the cancer is primary. In one embodiment, the cancer is metastatic. In one embodiment, the cancer is benign. In one embodiment, the cancer is malignant.

[0161] In one embodiment, the proliferative disorder to be treated and / or prevented is brain cancer. The brain cancer can be selected from: anaplastic astrocytoma, astrocytoma, central neurocytoma, choroid plexus carcinoma, choroid plexus papilloma, choroid plexus tumor, diffuse endocrine glioma, dysembryoplastic neuroepithelioma, ependymal tumor, fibrous astrocytoma, giant cell glioblastoma, multiforme glioblastoma, cerebroglioma, gliosarcoma, hemangiopericytoma, medulloblastoma, medullary epithelioma, meningeal carcinoma, neuroblastoma, neuroblastoma, oligodendrocyte glioma, ... The brain cancer is preferably a tumor (preferably a solid tumor). The brain cancer can be a primary cancer (for example, a glioma, a meningioma, a pituitary adenoma or a schwannoma) or a metastatic cancer (i.e., a brain cancer caused by a cancer in other parts of the body, such as a melanoma or a lung cancer).

[0162] Alternatively or additionally, the compounds may be administered in combination with other agents, such as chemotherapeutic or immunostimulatory drugs or therapeutic agents.

[0163] The terms "combination therapy" or "adjuvant therapy" in defining the use of a compound of the invention and one or more other agents are intended to include administration of each agent in a sequential manner in a regimen that provides the beneficial effects of the drug combination, and are also intended to include co-administration of these agents in a substantially simultaneous manner, such as in a single formulation having a fixed ratio of these active agents or in multiple separate formulations of each agent.

[0164] In various embodiments of the present invention, one or more compounds of formula (I) can be formulated or administered in combination with one or more other therapeutic agents. Therefore, in various embodiments of the present invention, one or more compounds of formula (I) can be included in a combined treatment regimen with surgery and / or other known treatments or therapeutic agents (such as other anticancer agents, particularly chemotherapeutic agents, radiotherapeutic agents and / or adjuvants or preventive agents).

[0165] There are a large number of available antitumor drugs in commercial use, clinical evaluation and preclinical development, which can be selected for the treatment of cancer or other tumors by combined drug chemotherapy. This antitumor drug is divided into several main categories, i.e., antibiotic type medicine, antimetabolite, hormone agent, immunizing agent, interferon type medicine and various medicines. Alternatively, other antitumor drugs (such as metalloproteinase inhibitors) can be used. Those skilled in the art will recognize the suitable medicine that can be used for combined therapy. Suitable reagents are listed in, for example, Merck Index, An Encyclopedia of Chemicals, Drugs and Biologicals, 12th edition, 1996.

[0166] The combination regimen may include the active agents being administered together, sequentially or spaced apart as appropriate in each case.The combination of active agents including the compounds of the invention may be synergistic.

[0167] The co-administration of the compound of formula (I) can be achieved by the compound of formula (I) and the chemotherapeutic agent or other anticancer agent in the same unit dose, or the compound of formula (I) and the chemotherapeutic agent or other anticancer agent can be present in separate and discrete unit doses, administered at the same or similar time. Sequential administration can be in any order desired, and when the second or subsequent compound is administered, it may be necessary to have a continued physiological effect of the first or initial compound, especially in cases where a cumulative or synergistic effect is desired.

[0168] For various applications, the compounds of the invention can be labeled with isotopes, fluorescent or luminescent markers, antibodies or antibody fragments, any other affinity markers (such as nanobodies, aptamers, peptides, etc.), enzymes or enzyme substrates. These labeled compounds of the invention are used to locate the position of receptors in vivo, in vitro, in vitro and in situ (e.g., in tissue sections) by autoradiography, and as radioactive tracers for positron emission tomography (PET) imaging, single photon emission computed tomography (SPECT), etc., to characterize those receptors in living or other materials. The labeled compounds of the invention can be used for treatment, diagnosis and other applications, such as in vivo and in vitro research tools, particularly the applications disclosed herein.

[0169] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more of the individual features mentioned or evident from the text or drawings. All these different combinations constitute various alternative aspects of the invention.

[0170] Embodiments of the present invention will now be discussed in more detail with reference to examples, which are provided for the purpose of illustration only and are not to be construed in any way as limiting the scope of the present invention.

[0171] Example

[0172] All chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA) except for boric acid which was purchased from Frontiers Scientific (Logan, UT, USA). Microwave irradiation was performed using a CEM-Discover microwave reactor (Kamp-Lintfort, Germany). The spectra were acquired on a BRUKER "Avance 300" 300 MHz NMR spectrometer (Bruker Corporation, Billerica, MA, USA). 1 H-NMR spectra. d6-DMSO or CDCl3 were obtained from Cambridge Isotope Laboratory. High performance liquid chromatography was performed on an Agilent Series 1200LC system equipped with an Agilent 1260 Infinity binary pump and an integrated vacuum degasser, autosampler, and diode array detector (Agilent Technologies, Santa Clara, California, USA). Samples were analyzed using a 0 to 100% gradient of acetonitrile: water over 40 minutes at a flow rate of 0.2 mL / min on an Agilent C18 column (particle size: 5 μM, 150×4.6 mm inner diameter). Eluted samples were analyzed using an Agilent 6120 quadrupole mass spectrometer. Agilent OpenLAB Chromatography Data System (CDS) ChemStation Edition was used for data acquisition and processing.

[0173] Primary antibodies against Bcl-2, PARP (#95425), secondary anti-rabbit (#7074), and anti-mouse (#7076) HRP-linked antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). EGFR (#sc03) and GAPDH (#sc737179) antibodies were from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against β-tubulin (#ab11308) were purchased from Abcam. Alexa488-conjugated anti-mouse secondary antibodies and DAPI (4',6'-diamidino-2-phenylindole) were from Life Technologies. CMPD1 (#sc-203138) was obtained from Santa Cruz Biotechnology. Paclitaxel and vinblastine were purchased from Sigma.

[0174] synthesis

[0175] See Scheme 1 for the preparation of the compounds.

[0176] 4-(4-Bromophenyl)-4-oxobutanoic acid

[0177] Succinic anhydride 3 (5.0 g, 50 mmol) and bromobenzene 2 (48 g, 300 mmol) were cooled to 0°C. Aluminum chloride (13.3 g, 100 mmol) was added and the mixture was stirred at 0°C under nitrogen for 4 hours. The reaction was allowed to warm to room temperature and stirred under nitrogen for 96 hours. The reaction was cooled to 0°C, concentrated HCl (125 mL) was added, and stirred under nitrogen for another hour. The reaction was filtered and washed with water (1 L) to give a light yellow solid, which was recrystallized from toluene to give 4-(4-bromophenyl)-4-oxobutanoic acid 4 (12 g, 46.68 mmol, 93.4%).

[0178] MS (ESI-quadrupole) m / z: C 10 Calculated for H9BrO3: 255.97, 257.97; Found: 258.0 (11), 257.0 (98), 256.0 (12), 255.0 (100), 213.0 (18), 211.0 (17) (negative ions).

[0179] HPLC:t r =24.4 minutes

[0180] 1H NMR (500MHz, d6-DMSO) δ: 2.59 (2H, t, J = 6.5 Hz, CH2), 3.21 (2H, t, J = 6.5 Hz, CH2), 7.88 (2H, d, J = 8.8 Hz, ArH), 7.96 (2H, d, J = 8.8 Hz, ArH), 12.19 (1H, br s, OH).

[0181] 4-(4-Bromophenyl)butyric acid 5

[0182] Zinc (13.0 g, 200 mmol) and mercuric chloride (1.00 g, 480 mmol) were stirred with water (10 mL) and concentrated HCl (0.6 mL) for 5 minutes. The liquid was poured off and toluene (20 mL), concentrated HCl (20 mL) and water (8 mL) were added successively. 4-(4-bromophenyl)-4-oxobutanoic acid 4 (2.55 g, 10.5 mmol) was added and heated to reflux at 100 °C for 24 hours, adding HCl (1 mL) every 6 hours. The reaction was cooled to room temperature, filtered and the solvent was removed from the organic layer to obtain a clear liquid, which gave white crystals after cooling. Purification by silica gel chromatography (ethyl acetate: hexane, 1:3) gave 4-(4-bromophenyl)butanoic acid 5 (2.33 g, 9.63 mmol, 91.4%).

[0183] MS (ESI-quadrupole) m / z: C 10 H 11 BrO2 calculated: 241.99, 243.99; found: 244.0 (10), 243.0 (98), 242.0 (11), 241.0 (100) (negative ion).

[0184] HPLC:t r =27.7 minutes

[0185] 1 H NMR (300 MHz, CDCl3) (δ / ppm) 7.40 (2H, d, J = 8.4 Hz, ArH), 7.06 (2H, d, J = 8.4 Hz, ArH), 2.63 (2H, t, J = 7.8 Hz, CH2), 2.36 (2H, t, J = 7.2 Hz, CH2), 1.97, (2H, dt, J = 7.2, 7.8 Hz, CH2), OH was not observed.

[0186] 4-(4-Bromophenyl)-N-(4-hydroxyphenyl)butyramide 7(1-17)

[0187] 4-(4-Bromophenyl)butyric acid 4 (0.5 g, 2.07 mmol) and benzotriazol-1-yl-oxytripyrrolidinophosphine hexafluorophosphate (PyBOP) (1.07 g, 2.07 mmol) were dissolved in dimethylformamide (10 mL). Diisopropylethylamine (710 μL, 4.14 mmol) was added and the reaction was stirred at 0°C for 30 minutes. 4-Aminophenol 6 (0.269 mg, 2.48 mmol) was added and the reaction was stirred at room temperature for 12 hours. The reactants were diluted with water (90 mL) and the product was extracted with dichloromethane (3×50 mL). The solvent was removed and the product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 35:65 to 60:40) to give 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 as a white solid (350 mg, 1.05 mmol, 51%).

[0188] MS (ESI-quadrupole) m / z: C 16 H 16 BrNO2 calculated: 333.04, 335.04; found: 334.0 (100), 335.0 (18), 336.0 (93), 337.0 (17).

[0189] HPLC:t r =28.2 minutes

[0190] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.59(1H,br s,NH),9.15(1H,br s,OH),7.47(2H,d,J=8.1Hz,ArH),7.33(2H,d,J=9.0Hz,ArH),7.18(2H,d,J=8.4Hz,ArH),6.67(2H,d,J =8.7Hz, ArH), 2.59 (2H, t, J = 7.8 Hz, CH2), 2.24 (2H, t, J = 7.2 Hz, CH2), 1.85 (2H, dt, J = 7.2, 7.8 Hz, CH2).

[0191] 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butyramide 10(1-18)

[0192]

[0193] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 (100 mg, 0.3 mmol), (2-fluoropyridin-3-yl)boric acid 8 (50.9 mg, 0.361 mmol), sodium carbonate (63.8 mg, 0.6 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4: 1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C with microwave irradiation (power adjustment 50 W start, pressure is not controlled) for 10 minutes. The reaction was cooled and the solvent was removed to obtain a yellow solid. The product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 45: 100 to 60: 40) to obtain 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butyramide 10 as an off-white solid (40 mg, 0.11 mmol, 38%).

[0194] MS (ESI-quadrupole) m / z: C 21 H 19 FN2O2 calculated: 350.14; found: 351.2 (100), 352.2 (24).

[0195] HPLC:t r =26.1 minutes.

[0196] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.61(1H,br s,NH),9.12(1H,br s,OH),8.23(1H,d,J=2.4Hz,ArH),8.09(1H,tt,J=8.4Hz,2.4,ArH),7.54(2H,d,J=8.4Hz,ArH),7.47(4H,d,J=8.7Hz,ArH),7.44(1H, d,J=4.2Hz,ArH), 6.68(2H,d,J=8.7Hz,ArH), 2.68(2H,t,J=7.8Hz,CH2), 2.30(2H,t,J=7.5Hz,CH2), 1.93(2H,dt,J=7.8,7.5Hz,CH2).

[0197] 4-(4-(2,6-difluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butyramide 12(1-22)

[0198]

[0199] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 (100 mg, 0.3 mmol), (2,6-difluoropyridin-3-yl)boric acid 11 (50.9 mg, 0.36 mmol), sodium carbonate (63.8 mg, 0.6 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4:1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C with microwave irradiation (power adjustment 50 W initial, pressure uncontrolled) for 10 minutes. The reaction was cooled and the solvent was removed to obtain a yellow solid. The product was purified by silica gel chromatography (ethyl acetate:hexanes, gradient 25:100 to 50:50) to afford 4-(4-(2,6-difluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butanamide 12 as an off-white solid (30 mg, 0.08 mmol, 27%).

[0200] MS (ESI-quadrupole) m / z: C 21 H 18 Calculated for F2N2O2: 368.13; Found: 369.1(100), 370.1(23).

[0201] HPLC:t r =28.6 minutes.

[0202] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.62(1H,br s,NH),9.17(1H,br s,OH),8.29(1H,dt,J=7.5,5.0Hz,ArH),7.52(2H,d,J=6.6Hz,ArH),7.35(4H,m,ArH),7.27(1H,dd,J=8.4,2.7Hz,Ar H), 6.67 (2H, d, J = 9Hz, ArH), 2.67 (2H, t, J = 7.8Hz, CH2), 2.29 (2H, t, J = 7.5Hz, CH2), 1.91 (2H, dt, J = 7.8, 7.5Hz, CH2).

[0203] 4-(4-(6-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butyramide 14(1-21)

[0204]

[0205] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 (100 mg, 0.299 mmol), (6-fluoropyridin-3-yl)boric acid 13 (50.96 mg, 0.359 mmol), sodium carbonate (63.8 mg, 0.600 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4: 1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C with microwave irradiation (power adjustment 50 W initial, pressure uncontrolled) for 10 minutes. The reaction was cooled and the solvent was removed to obtain a yellow solid. The product was purified by silica gel chromatography (ethyl acetate:hexanes, gradient 25:100 to 50:50) to afford 4-(4-(6-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butanamide 14 as an off-white solid (30 mg, 0.09 mmol, 29%).

[0206] MS (ESI-quadrupole) m / z: C 21 H 19 FN2O2 calculated: 350.14; found: 351.1 (100), 352.1 (23).

[0207] HPLC:t r =26.8 minutes.

[0208] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.61(1H,br s,NH),9.16(1H,br s,OH),8.52(1H,d,J=2.4Hz,ArH),8.25(1H,tt,J=8.4Hz,2.4,ArH),7.64(2H,d,J=8.4Hz,ArH),7.34(4H,d,J=8.7Hz,ArH),7.26(1H,dd, J=4.2,3.0Hz,ArH),6.66(2H,d,J=8.7Hz,ArH),2.66(2H,t,J=7.8Hz,CH2),2.28(2H,t,J=7.5Hz,CH2),1.90(2H,dt,J=7.8,7.5Hz,CH2).

[0209] N-(4-Hydroxyphenyl)-4-(4-(pyridin-3-yl)phenyl)butyramide 16(1-38)

[0210]

[0211] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 (100 mg, 0.3 mmol), pyridin-3-ylboronic acid 15 (67.1 mg, 0.36 mmol), sodium carbonate (63.8 mg, 0.6 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4:1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C for 10 minutes with microwave irradiation (power adjustment 50 W start, pressure uncontrolled). The reaction was cooled and the solvent was removed to give a yellow solid. The product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 25: 100 to 50: 50) to give N-(4-hydroxyphenyl)-4-(4-(pyridin-3-yl)phenyl)butyramide 16 as an off-white solid (40 mg, 0.11 mmol, 35%).

[0212] MS (ESI-quadrupole) m / z: C 21 H 20 N2O2 calculated: 332.15; found: 333.2 (100), 334.1 (23).

[0213] HPLC:t r =17.3 minutes.

[0214] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.62(1H,br s,NH),9.16(1H,br s,OH),8.88(1H,s,ArH),8.55(1H,d,J=4.5Hz,ArH),8.05(1H,d,J=7.8Hz,ArH),7.66(2H,d,J=7.8Hz,ArH),7.48(1H,q,J=4.8Hz,ArH), 7.35(4H,d,J=8.4Hz,ArH), 6.67(2H,d,J=8.7), 2.68(2H,t,J=6.9Hz,CH2), 2.29(2H,t,J=7.5Hz,CH2), 1.92(2H,dt,J=6.9,7.5Hz,CH2).

[0215] N-(4-Hydroxyphenyl)-4-(4-(2-methoxypyrimidin-5-yl)phenyl)butyramide 18(1-19)

[0216]

[0217] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide 7 (100 mg, 0.299 mmol), (2-methoxypyrimidin-5-yl)boronic acid 17 (55.25 mg, 0.359 mmol), sodium carbonate (63.8 mg, 0.600 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4: 1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C with microwave irradiation (power adjustment 50 W initial, pressure uncontrolled) for 10 minutes. The reaction was determined to have low conversion by thin layer chromatography, so it was heated for another 20 minutes with microwave irradiation as described above. The solvent was removed and the product was purified by silica gel chromatography (ethyl acetate:hexanes, gradient 25:100 to 50:50) to give 4-(4-(6-fluoropyridin-3-yl)phenyl)-N-(4-hydroxyphenyl)butanamide 18 as an off-white solid (3 mg, 0.08 mmol, 3%).

[0218] MS (ESI-quadrupole) m / z: C 21 H 21 N3O3 calculated: 363.16; found: 364.2 (100), 365.2 (24).

[0219] HPLC:t r =24.3 minutes.

[0220] 4-(4-Bromophenyl)-N-phenylbutyramide 20

[0221]

[0222] 4-(4-Bromophenyl)butyric acid 5 (0.5 g, 2 mmol) and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) (1.07 g, 2.07 mmol) were dissolved in dimethylformamide (10 mL). Diisopropylethylamine (710 μL, 4.14 mmol) was added and the reaction was stirred at 0 °C for 30 minutes. Aniline 19 (225.0 μL, 2.47 mmol) was added and the reaction was stirred at room temperature for 12 hours. The reaction was then diluted with water (90 mL) and the product was extracted with dichloromethane (3×50 mL). The solvent was removed and the product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 20:80 to 40:60) to give 4-(4-bromophenyl)-N-phenylbutyramide 20 as a white solid (500 mg, 1.57 mmol, 76%).

[0223] MS (ESI-quadrupole) m / z: C 16 H 16BrNO calculated: 317.04, 319.04; found: 318.1 (100), 319.1 (19), 320.1 (96), 321.1 (17), 340.0 (33), 341.0 (6), 342.0 (33), 343.0 (5).

[0224] HPLC:t r =35.7 minutes.

[0225] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.85(1H,br s,NH),7.57(2H,d,J=7.5Hz,ArH),7.46(2H,d,J=8.4Hz,ArH),7.27(2H,t,J=8.1Hz,ArH),7.18(2H,d,J=8.4Hz,ArH), 7.01(1H,t,J=7.2Hz,ArH),2.60(2H,t,J=7.2Hz,CH2),2.30,(2H,t,J=7.5Hz,CH2),1.87(2H,dt,J=7.2,7.5Hz,CH2).

[0226] 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-phenylbutyramide 21(1-20)

[0227]

[0228] 4-(4-bromophenyl)-N-phenylbutanamide 20 (100 mg, 0.314 mmol), (2-fluoropyridin-3-yl)boric acid 8 (52.8 mg, 0.377 mmol), sodium carbonate (66.7 mg, 0.628 mmol) and tetrakis(triphenylphosphine)palladium(0) (17 mg, 0.015 mmol) were dissolved in ethanol: water (4:1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C for 10 minutes with microwave irradiation (power adjustment 50 W start, pressure uncontrolled). The reaction was cooled and the solvent was removed to give a yellow solid. The product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 30: 100 to 60: 40) to give 4-(4-(2-fluoropyridin-3-yl)phenyl)-N-phenylbutanamide 21 as a yellow solid (30 mg, 0.11 mmol, 38%).

[0229] MS (ESI-quadrupole) m / z: C 21 H 19 FN2O: 334.15; found 335.2 (100), 336.2 (23), 357.2 (14).

[0230] HPLC:t r=31.1 minutes.

[0231] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.85(1H,br s,NH),8.22(1H,d,J=4.8Hz,ArH),8.09(1H,ddd,J=10.2,4.8,1.8Hz,ArH),7.56(4H,m,ArH),7.36(2H,d,J=8.4Hz,ArH),7.28(2H,t ,J=8.4Hz,ArH),7.01(1H,t,J=7.2Hz,ArH),2.68(2H,t,J=7.5Hz,CH2),2.35,(2H,t,J=7.5Hz,CH2),1.94(2H,q,J=7.5,7.5Hz,CH2).

[0232] N-(4-Hydroxyphenyl)-4-(4-(pyrimidin-5-yl)phenyl)butanamide (1-01)

[0233]

[0234] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butyramide (100 mg, 0.3 mmol), pyrimidin-5-ylboronic acid (44.8 mg, 0.361 mmol), sodium carbonate (64 mg, 0.6 mmol) and tetrakis(triphenylphosphine)palladium(0) (21 mg, 0.018 mmol) were dissolved in ethanol: water (4: 1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 ° C for 10 minutes with microwave irradiation (power adjustment 50 W start, pressure uncontrolled). The reaction was cooled and the solvent was removed to give a yellow solid. The product was purified by silica gel chromatography (ethyl acetate: hexane, gradient 35: 75 to 85: 15) to give N-(4-hydroxyphenyl)-4-(4-(pyrimidin-5-yl)phenyl)butyramide as an off-white solid (20 mg, 0.06 mmol, 20%).

[0235] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.61(1H,br s,NH),9.16(1H,br s,OH),9.13(3H,m,ArH),7.74(2H,d,J=8.1Hz,ArH),7.38(4H,m,ArH),6.66(2H,d,J= 8.7Hz, ArH), 2.68 (2H, t, J = 7.2 Hz, CH2), 2.29 (2H, t, J = 7.2 Hz, CH2), 1.92 (2H, m, CH2).

[0236] 4-(4'-(Aminomethyl)-[1,1'-biphenyl]-4-yl)-N-(4-hydroxyphenyl)butyramide (1-07)

[0237]

[0238] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butanamide (101 mg, 0.3 mmol), (4-(aminomethyl)phenyl)boric acid (71 mg, 0.38 mmol), potassium tert-butoxide (75 mg, 0.45 mmol) and tetrakis(triphenylphosphine)palladium(0) (27 mg, 0.02 mmol) were dissolved in ethanol:water (4:1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 130 °C for 30 minutes with microwave irradiation (power adjustment 50 W starting, pressure not controlled). The reaction was cooled and the solvent was removed to give a yellow solid. The product was purified by silica gel chromatography (methanol: dichloromethane, gradient 5:95 to 15:85) to give 4-(4'-(aminomethyl)-[1,1'-biphenyl]-4-yl)-N-(4-hydroxyphenyl)butanamide as an off-white solid (30 mg, 0.08 mmol, 30%).

[0239] 1 H NMR(300MHz,d6-DMSO)(δ / ppm)9.62(1H,br s,OH),7.57(4H,m,ArH),7.34(6H,m,ArH),6.67(2H,d,J=8.7Hz,ArH),3.75(2H,s , CH2), 2.64(2H,t,J=7.5Hz,CH2), 2.28(2H,t,J=7.5Hz,CH2), 1.896(2H,m,CH2).

[0240] N-(4-Hydroxyphenyl)-4-(4-(quinolin-3-yl)phenyl)butanamide (1-08)

[0241]

[0242] 4-(4-bromophenyl)-N-(4-hydroxyphenyl)butanamide (105 mg, 0.31 mmol), quinolin-3-ylboronic acid (65 mg, 0.38 mmol), sodium carbonate (55 mg, 0.52 mmol) and tetrakis(triphenylphosphine)palladium(0) (21 mg, 0.018 mmol) were dissolved in ethanol:water (4:1 v / v, 2 mL). The solution was purged with nitrogen and heated in a sealed tube at 120 °C with microwave irradiation (power adjustment 50 W starting, pressure not controlled) for 60 minutes. The reaction was cooled and the solvent was removed to give a yellow solid. The product was purified by flash silica gel chromatography (ethyl acetate: hexane, gradient 0: 100 to 100: 0 over 60 minutes) to give N-(4-hydroxyphenyl)-4-(4-(quinolin-3-yl)phenyl)butanamide as an off-white solid (30 mg, 0.08 mmol, 25%).

[0243] 1 H NMR(400MHz,d6-DMSO)(δ / ppm)9.61(1H,br s,NH),9.25(1H,s,ArH),9.11(1H,br s,OH),8.62(1H,s,ArH),8.05(2H,d,J=8.4Hz,ArH),7.82(2H,d,J=8.4Hz,ArH),7.77(1H,t,J=8.0Hz,ArH),7.61(1H,m,Ar H),7.38(4H,m,ArH),6.67(2H,d,J=8.8Hz,ArH),2.70(2H,t,J=7.6Hz,CH2),2.31(2H,t,J=7.6Hz,CH2),1.95(2H,m,CH2).

[0244] Biological evaluation

[0245] U87 and U251 cell lines are obtained from European cell line / microorganism collection center (European Collection of Cell Cultures) (ECACC) by Australian cell bank (CellBank).U87-EGFRvIII cell line (Nishikawa et al., 1994, Proc Natl Acad Sci USA 91:7727-7731) is obtained from its native laboratory, and EGFR amplification and sudden change are detected by immunoblotting and flow cytometry.Cell is cultivated at 37 ℃ and 5%CO2 in the DMEM culture medium (Invitrogen byLife Technologies, Carlsbad, California, the U.S.) supplemented with 10%FBS (Sigma-Aldrich). Primary glioma spheres (RN1, classical; WK1, mesenchymal) were derived from glioblastoma samples (Day et al., 2013, Cancer 5:357-371; Day et al., 2013, Cancer Cell 23:238-248), maintained in StemProNSC serum-free medium supplemented with EGF and FGF (both Life Technologies), and grown on Matrigel-coated flasks (Pollard et al., 2009, Cell Stem Cell 4:568-580).

[0246] EC of the compounds of the present invention 50 value.

[0247] Cell viability was determined using the Alamar blue assay after 72 h of treatment (Table 2). EC was calculated using nonlinear regression on Graph Pad Prism version 5. 50 cLogP values ​​were calculated using ChemBioDraw software. Results are the mean ± SEM of three independent experiments performed in triplicate.

[0248] Table 2. EC of compounds of the present invention 50 value

[0249]

[0250]

[0251] Further analysis

[0252] like Figure 1As shown, SC-38 reduces the viability of glioma cell lines and is selectively cytotoxic. Cells were treated with SC-38 (10 nM-10 μM) for 72 hours. Cells were tested for viability (left, Alamar Blue viability assay, Invitrogen) and supernatants were harvested for bioluminescent cytotoxicity analysis (right, ToxiLight TM Bioassay kit, Lonza).

[0253] like Figure 2 As shown, SC-38 is selectively toxic to malignant cells. Cells were treated with SC-38 (1 μM) for 72 hours and subjected to viability assays (Alamar blue, MTT or PI staining) to determine the viability of malignant (U87, primary glioma) and non-malignant (microglia, neurons, astrocytes, macrophages) cells.

[0254] like Figure 3 As shown, compounds 1-18 readily cross the BBB. BALB / c mice were intraperitoneally injected with the tested inhibitors (10 mg / kg), and the brains were collected at 30 and 60 minutes. The brain homogenates were used for solid phase extraction, and the amount of the tested compounds was determined by HPLC. Lapatinib data are from Polli, JW et al., “The Role of Efflux and Uptake Transporters in N-{3-Chloro-4-[(3-fluorobenzyl)oxy]phenyl}-6-[5-({[2-(methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine (GW572016, Lapatinib) Disposition and Drug Interactions,” Drug Metabolism and Disposition 36, 695-701 (2008).

[0255] like Figure 4 As shown, SC-38 induced apoptosis in U87 and U87vIII cells. Figure 4 ,AB) and U87vIII( Figure 4, CD) cells were treated with SC-38 for 48 hours and stained with Annexin-FITC / 7-AAD. Apoptotic cells were visualized using MUSE cell analyzer (Millipore). Results are mean ± SEM of 3 independent experiments. Statistical significance was calculated using one-way ANOVA and Dunnet post-test (**P<0.01, ***P<0.001).

[0256] Western blot analysis of PARP cleavage in U87 and U87vIII cells treated with SC-38 was also performed (see Figure 5 ). U87 (lanes 1-3) and U87vIII (lanes 4-6) cells were treated with 1 and 5 μM SC-38 for 48 hours. Representative blots of 3 independent experiments are shown ( Figure 5 A). GAPDH was used as a loading control. PARP cleavage was quantified and normalized to GAPDH (expressed as fold change), see Figure 5 B. Results are mean ± SEM from 3 independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnet's post-test (*P<0.05, ***P<0.001, ****P<0.0001).

[0257] like Figure 6 As shown, compound 1-08 induced apoptosis in U87 and U87vIII cells. Figure 6 ,AB) and U87vIII( Figure 6 , CD) cells for 48 hours and stained with Annexin-FITC / 7-AAD. Apoptotic cells were visualized using MUSE cell analyzer (Millipore). Results are mean ± SEM from three independent experiments. Statistical significance was calculated using one-way ANOVA and Dunnet post-test (***P<0.001).

[0258] After U87 and U87vIII cells were treated with compound 1-08, Western blot analysis of PARP cleavage in cells was also performed ( Figure 7 ). U87 (lanes 1-3) and U87vIII (lanes 4-6) cells were treated with 1 and 5 μM compound 1-08 for 48 hours. Representative blots of 3 independent experiments are shown (see Figure 7 A). GAPDH was used as a loading control. PARP cleavage was quantified and normalized to GAPDH (expressed as fold change), see Figure 7 B. Results are mean ± SEM from 3 independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnet's post-test (*P<0.05, ****P<0.0001).

[0259] The effects of SC-38 treatment on Bcl-2 family proteins in U87, U87vIII, and U251 were also tested (see Figure 8 ). U87 (lanes 1-3), U87vIII (lanes 4-6), and U251 (lanes 7-9) cells were treated with SC-38 (1 and 5 μM) for 24 hours and subjected to Western blot analysis, using GAPDH as a loading control. Representative blots are shown in Figure 8 A, and the quantification normalized to GAPDH is shown in Figure 8 In BE, shown as fold change. Results are mean ± SEM from three independent experiments, and statistical significance was determined using unpaired t-test (*P<0.05, **P<0.01, ****P<0.0001).

[0260] The effects of compound 1-08 treatment on Bcl-2 family proteins in U87, U87vIII, and U251 were also investigated (see Fig. 9 ). U87 (lanes 1-3), U87vIII (lanes 4-6), and U251 (lanes 7-9) cells were treated with compound 1-08 (1 and 5 μM) for 24 hours and subjected to Western blot analysis, where GAPDH was used as a loading control. Representative blots are shown in Fig. 9 A, and the quantification is shown in Fig. 9 BE, relative protein expression is shown. Results are mean ± SEM from 2 independent experiments, and statistical significance was determined using unpaired t-test (*P < 0.05, **P < 0.01).

[0261] Kinase profiles of compound 1-08 at 10 and 50 μM were performed by ProQinase (Fribourg, Germany). Activity assay) and non-radioactive ADP-Glo TM The assay (Promega, Madison, Wisconsin, USA) measured the kinase activity of 387 protein kinases and 13 lipid kinases, respectively. The results are shown in Table 3 (only the top 10 kinases are listed, and the selectivity score describes the fraction of kinases that inhibited more than 50% among all tested kinases).

[0262] Table 3. Kinase group results of compound 1-08

[0263]

[0264] At 10 μM, compound 1-08 inhibited MERTK at a level of 51%. Several other kinases were inhibited at a concentration of 50 μM.

[0265] The effect of compound 1-08 on tubulin polymerization and mitotic spindle formation was also studied. Fluorescence-based tubulin polymerization assays were performed in a final volume of 55 μL using a tubulin polymerization assay kit (Cytoskeleton, Colorado, USA) according to the manufacturer's instructions. Porcine brain tubulin was incubated with test compounds (paclitaxel, vinblastine, and compound 1-08) at 37°C and fluorescence was measured using a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany; excitation at 355 nm and emission at 460 nm). Paclitaxel enhanced tubulin polymerization, while vinblastine and compound 1-08 inhibited tubulin polymerization in a dose-dependent manner compared to the control ( Fig.10 A). To test whether compound 1-08 alters microtubules in vivo, U87 glioblastoma cells were treated with compound 1-08 and the effect on microtubules was determined by immunofluorescence staining of β-tubulin. Treatment of U87 cells with compound 1-08 (1 and 5 μM) resulted in the disassembly of microtubules ( Fig.10 B) and a dose-dependent increase in the number of multinucleated cells ( Fig.10 B and 10C). The consequence of microtubule depolymerization induced by compound 13 is disrupted spindle assembly ( Fig.10 D). In untreated U87 cells undergoing mitosis, microtubules assemble into bipolar spindles (control figure). However, treatment with compound 1-08 results in defective spindle formation and chromosome misalignment. These results indicate that compound 1-08 is a tubulin depolymerizing agent that disrupts mitotic spindle formation and causes multinucleated cell formation.

[0266] The effects of SC-38 and compounds 1-08, 1-18, and 1-38 were also investigated in a cell viability assay using patient-derived glioma spheres. RN1 glioma spheres, representing a classical subtype of glioblastoma, were the most responsive to treatment ( Fig.11 , wherein SC-38 is referred to as "CMPD1", compound 1-08 is referred to as "compound 13", compound 1-18 is referred to as "compound 7", and compound 1-38 is referred to as "compound 10"). SC-38 and compound 1-08 attenuated the viability of RN1 glioma spheres, EC 50 0.26 and 0.54 μM respectively ( Fig.11 A). SC-38 also effectively attenuated the growth of mesenchymal WK1 glioma spheres (EC 50 =0.91 μM, Fig.11 B). In mesenchymal glioma spheres, when combined with compound 1-08 (EC 50 =3.82 μM), compound 1-18 was more effective (EC50 =2μM)。

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt or prodrug thereof, wherein: X is a C1-C6 alkyl group or a C2-C6 alkenyl group; Y is CH2, NH, N-alkyl, N-alkenyl, S or O; W is O, S or NH; Z is CH2, NH, N-alkyl, N-alkenyl, S or O; R1 is halogen, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein the cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted; Ar is aryl or heteroaryl; R2 is H, OH, NH2 or NO2.

2. The compound according to claim 1, characterized in that X is a C2 alkyl group or a C2 alkenyl group.

3. The compound according to claim 1 or 2, characterized in that Y is CH2.

4. A compound according to any one of the preceding claims, characterized in that Z is NH.

5. The compound according to any one of claims 1 to 3, characterized in that Z is O.

6. The compound according to claim 1 or 2, characterized in that Y is NH.

7. The compound according to claim 1 or 2, characterized in that Y is O.

8. The compound according to claim 1, 2, 6 or 7, characterized in that Z is CH2.

9. The compound according to claim 1 or 2, characterized in that Y and Z are both NH.

10. A compound according to any one of the preceding claims, characterized in that W is O.

11. A compound according to any one of the preceding claims, characterized in that R1 is halogen.

12. The compound according to claim 11, characterized in that The halogen is Br.

13. The compound according to any one of claims 1 to 10, characterized in that R1 is aryl.

14. The compound according to claim 13, characterized in that The aryl group is monocyclic or bicyclic.

15. The compound according to claim 13 or 14, characterized in that The aryl group is phenyl or naphthyl.

16. The compound according to any one of claims 13 to 15, characterized in that The aryl group is substituted.

17. The compound according to claim 16, characterized in that The substituents are selected from halogen and heteroalkyl.

18. The compound according to claim 17, characterized in that The halogen is F.

19. The compound according to claim 17, characterized in that The heteroalkyl group is an O-alkyl group.

20. The compound according to claim 19, characterized in that O-alkyl is -OCH3.

21. The compound according to claim 17, characterized in that The heteroalkyl group is an aminoalkyl group.

22. The compound according to claim 21, characterized in that Aminoalkyl is -CH2NH2.

23. The compound according to any one of claims 1 to 10, characterized in that R1 is heteroaryl.

24. The compound according to claim 23, characterized in that The heteroaryl group is monocyclic or bicyclic.

25. The compound according to claim 23 or 24, characterized in that The heteroaryl group includes one or more nitrogen atoms.

26. The compound according to claim 25, characterized in that The heteroaryl group is pyrazole, isoxazole, triazole, pyridine, pyrimidine, quinoline, benzimidazole or indole.

27. The compound according to any one of claims 23 to 26, characterized in that The heteroaryl group is substituted.

28. The compound according to claim 27, characterized in that The substituents are selected from halogen and heteroalkyl.

29. The compound according to claim 28, characterized in that The halogen is F.

30. The compound according to claim 28, characterized in that The heteroalkyl group is an O-alkyl group.

31. The compound according to claim 30, characterized in that O-alkyl is -OCH3.

32. The compound according to claim 28, characterized in that The heteroalkyl group is an aminoalkyl group.

33. The compound according to claim 32, characterized in that Aminoalkyl is -CH2NH2.

34. The compound according to any one of claims 1 to 10, characterized in that R1 is a heterocycloalkyl group.

35. The compound according to claim 34, characterized in that The heterocycloalkyl group includes one or more nitrogen atoms.

36. The compound according to claim 35, characterized in that The heterocycloalkyl group is piperazine.

37. The compound according to claim 34 or 35, characterized in that The heterocycloalkyl group includes one or more oxygen atoms.

38. The compound according to claim 37, characterized in that The heterocycloalkyl group is morpholine.

39. The compound according to any one of claims 34 to 38, characterized in that The heterocycloalkyl group is substituted.

40. The compound according to claim 39, characterized in that The substituents are selected from halogen and heteroalkyl.

41. The compound according to claim 40, characterized in that The halogen is F.

42. The compound according to claim 40, characterized in that The heteroalkyl group is an O-alkyl group.

43. The compound according to claim 42, characterized in that O-alkyl is -OCH3.

44. The compound according to claim 40, characterized in that The heteroalkyl group is an aminoalkyl group.

45. The compound of claim 44, wherein Aminoalkyl is -CH2NH2.

46. ​​A compound as claimed in any one of the preceding claims, characterized in that Ar is aryl.

47. The compound of claim 46, wherein The aryl group is phenyl.

48. A compound according to any one of claims 1 to 45, characterized in that Ar is heteroaryl.

49. The compound of claim 48, wherein The heteroaryl group includes one or more nitrogen atoms.

50. The compound according to claim 48 or 49, characterized in that The heteroaryl group has 4 or 5 ring carbon atoms.

51. A compound according to any one of claims 48 to 50, characterized in that The heteroaryl group is pyridine or pyrimidine.

52. A compound as claimed in any preceding claim, characterized in that R2 is H or OH.

53. A compound as claimed in any one of the preceding claims, characterized in that R2 is in the counter position.

54. A pharmaceutical composition comprising a compound of formula (I) as claimed in any one of the preceding claims, and a pharmaceutically acceptable carrier, diluent or excipient.

55. A method for treating or preventing a proliferative disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) as described in any one of claims 1-53.

56. A method of treating or preventing a proliferative disease in a subject, comprising administering to the subject the pharmaceutical composition of claim 54.

57. The method of claim 55 or 56, wherein: The proliferative disease is cancer.

58. The method of claim 57, wherein: The cancer is brain cancer.

Images