Selective CDK4 / 6 inhibitor-type cancer treatment drugs

Abstract

The present disclosure describes selective and potent CDK 4 / 6 inhibitors that show favorable inhibition of cancer growth even at low concentrations. Such anticancer CDK 4 / 6 inhibitors are substituted pyrrolopyrimidine compounds of Formula 1A having a fatty acid moiety. These compounds can be used as pharmaceutical compounds for anticancer therapy and can be used to treat, prevent and / or ameliorate cancer.

Description

[0001] field

[0002] The present disclosure relates to pharmaceutical compounds for anti-cancer treatment, and more particularly, to substituted pyrrolopyrimidine compounds, substituted pyrrolopyrimidine compounds and substituted benzimidazole compounds, which are effective CDK 4 / 6 inhibitors for treating, preventing and / or ameliorating cancer.

[0003] background

[0004] Cancer stem cells (CSCs) are tumor-initiating cells (TICs) that are resistant to conventional cancer treatments such as chemotherapy and radiotherapy. As a result, CSCs contribute to tumor recurrence and distant metastasis, leading to treatment failure and poor clinical outcomes in cancer patients. Therefore, innovative approaches are needed to understand how to address the CSC problem. Mechanistically, this may be related to the ability of CSCs to survive and thrive under harsh conditions and different microenvironments. Because CSCs are a particularly small subpopulation of the tumor cell population, their metabolic and phenotypic properties have remained largely uncharacterized until recently.

[0005] In addition, CSC has amazing resilience and high resistance to cell stress, which enables it to carry out anchorage-independent growth, especially under low attachment conditions. Therefore, it forms a 3D sphere, which retains the properties of CSC and stem cell progenitor cells. On the contrary, when suspended growth is carried out, most "main body" cancer cells die by anoikis (a special type of apoptosis). Therefore, the clonal propagation of a single CSC will produce a 3D sphere, and the self-aggregation of cancer cells is not involved. Therefore, 3D sphere formation is a functional readout of stemness in epithelial cancer cells, and allows enrichment of epithelial cell colonies with stem cell-like phenotypes. When these 3D spheres are prepared using breast cancer cells (such as MCF7, etc.), these 3D spheres are also referred to as mammary spheres.

[0006] Previously, 3D spheroids had been generated from 2 different ER(+) cell lines (MCF7 and T47D) and subjected to unbiased, label-free proteomic analysis. This work set out to analyze the phenotypic response of CSCs at the molecular level. 3D spheroids were directly compared to monolayers of these cell lines and processed in parallel. This allowed identification of the proteomic profile of 3D spheroids relative to monolayers, i.e., the characteristic CSC phenotype. Based on this molecular analysis, it was observed that mammospheres were greatly enriched for mitochondrial proteins. These mitochondrial-associated proteins include molecules involved in β-oxidation and ketone metabolism / recycling, mitochondrial biogenesis, electron transport, ADP / ATP exchange / transport, CoQ synthesis and ROS generation, as well as inhibition of mitophagy. Therefore, increasing mitochondrial protein synthesis or reducing mitophagy may allow mitochondrial mass accumulation in CSCs.

[0007] Given the increase in CSCs, mitochondrial mass was considered as a novel metabolic biomarker for purifying CSCs. Using this holistic approach, it has been observed that CSC activity can be greatly enriched using only MitoTracker as a single marker for both ER(+) (MCF7) and ER(-) (MDA-MB-231) breast cancer cell lines. Notably, high MitoTracker cells were found to be chemoresistant to paclitaxel and showed resistance to paclitaxel-induced DNA damage response.

[0008] However, what is needed are new pharmaceutical compounds for anticancer therapy that eradicate CSCs, prevent or reduce the likelihood of metastasis and / or recurrence, and reduce or eliminate cancer resistance to chemotherapy and other anticancer treatments. In addition, what is needed are therapeutic strategies and anticancer therapies that target the "optimally grown" CSCs and eliminate further cancer growth, including anchorage-independent growth, tumor recurrence, and distant metastasis.

[0009] Brief Overview

[0010] Cancer stem cells (CSCs) are currently considered one of the main sources of treatment failure for cancer patients worldwide. Mechanistically, this may be related to the ability of CSCs to survive and thrive under harsh conditions and different microenvironments. The inventors have proposed a theory that CSCs may become resistant to traditional therapies by "boosting" ATP production using elevated mitochondrial OXPHOS metabolism. Consistent with this view, a variety of mitochondrial inhibitors successfully blocked the formation of 3D tumor spheroids, including, in particular, i) FDA-approved antibiotics (doxycycline, tigecycline, azithromycin, embovium, atovaquone, bedaquiline), ii) natural compounds (actinonin, CAPE, berberine, brutieridin, and melittin), and iii) experimental compounds (oligomycin and AR-C155858, MCT1 / 2 inhibitors), etc.

[0011] Cyclin-dependent kinases (CDKs) 4 and 6 are enzymes known to promote cell mitosis and meiosis in normal and cancer cells. These enzymes are responsible for phosphorylating and inactivating the retinoblastoma protein, which plays a role in the progression of the cell cycle from the G1 phase to the S phase. Studies have found that abnormalities in cancer cells increase the activity of CDKs. This increased activity leads to the inactivation of various tumor suppressor genes, paving the way for the rapid proliferation of cancer stem cells and tumor growth. Naturally occurring protein inhibitors of CDKs, such as p16 and p27, have been shown to inhibit the growth of lung cancer cell lines in vitro. Certain CDK inhibitors can be used as chemoprotectants through their ability to inhibit the cell cycle progression of normal, non-transformed cells.

[0012] Targeted inhibition of these enzymes is a potential strategy for anticancer therapy and therapeutic drugs, whether used alone or in combination with other therapies. Blocking the CDK 4 / 6 pathway prevents cells from progressing to the S phase, which achieves cell death by apoptosis. Three classes of CDK inhibitors are described herein, mainly CDK 4 and CDK 6 inhibitors ("CDK 4 / 6"), which have strong therapeutic effects as cancer therapeutic agents. The first class of anticancer CDK 4 / 6 inhibitors are substituted pyrrolopyrimidine compounds with fatty acid moieties. The formula shown below is an example of some embodiments of the first class of anticancer CDK 4 / 6 inhibitors, wherein "n" is an integer of 9-20, more preferably 12-20.

[0013]

[0014] The second class includes substituted pyridopyrimidines having fatty acid moieties. The formula shown below is illustrative of some embodiments of the second class of anticancer CDK 4 / 6 inhibitors, wherein "n" is an integer of 9-20, more preferably 12-20.

[0015]

[0016] The third class includes substituted benzimidazole compounds having a fatty acid moiety. The formula shown below is an illustration of an embodiment of the third class of anticancer CDK 4 / 6 inhibitors, wherein "m" is an integer of 0-4, more preferably 0-2, and "n" is an integer of 9-20, more preferably 12-20.

[0017]

[0018] The first, second or third class compounds, including their salts, can be used as pharmaceutical compounds for the treatment of cancer. Exemplary salts include, in particular, succinate, trifluoroacetate, tartrate and malate, and other salts that will be understood by those of ordinary skill in the art. The present method also provides a pharmaceutical preparation having a therapeutically effective amount of a compound from the first, second or third class, or in some embodiments from one or more compounds of each class, or its therapeutically acceptable salt (multiple), and its pharmaceutically acceptable carrier, diluent or excipient. All of these forms are within the scope of the current method. It should be understood that pharmaceutically acceptable carriers known in the art can be used.

[0019] The compounds described herein can be used in conjunction with a method of treating cancer in a mammal (including a human), comprising administering to the mammal an amount of a compound from the first, second, or third class, or a pharmaceutically acceptable salt thereof, that is effective in treating such a disorder or condition. For example, the present method can be used to treat abnormal cell proliferation, such as cancer. The compounds described herein are useful for treating abnormal cell proliferation disorders, particularly cancers selected from the group consisting of breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, esophageal cancer, gastric cancer, skin cancer, lung cancer, bone cancer, colon cancer, pancreatic cancer, thyroid cancer, bile duct cancer, buccal cancer and pharyngeal (oral) cancer, lip cancer, tongue cancer, oral cancer, pharyngeal cancer, small intestine cancer, colorectal cancer, large intestine cancer, rectal cancer, brain and central nervous system, glioblastoma, neuroblastoma, keratoacanthoma, epidermoid carcinoma, large cell carcinoma, adenocarcinoma, adenocarcinoma, adenocarcinoma, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver cancer, kidney cancer, myeloid diseases, lymphatic system diseases, Hodgkin's disease, hairy cell and leukemia, by administering to a subject diagnosed with such a cancer a therapeutically effective amount of a compound from the first, second or third class, or a pharmaceutically acceptable salt thereof. In some embodiments, the present methods may be used in conjunction with other therapies and / or to enhance the effectiveness of other therapies.

[0020] Some embodiments of the present methods may take the form of compounds having the general formula,

[0021] in:

[0022] R 4 is selected from the group consisting of hydrogen, C1-C8-alkyl, substituted C1-C8-alkyl, C3-C8-cycloalkyl, substituted C3-C8-cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl;

[0023] Z is CR z , where R z is selected from halogen, hydrogen, C1-C3-alkyl, C1-C3-alkoxy, CN, C=NOH, C=NOCH3, C(O)H, C(O)C1-C3-alkyl, C3-C8-cycloalkyl, heterocyclyl, aryl, heteroaryl, substituted C1-C3-alkyl, substituted C3-C8-cycloalkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl, -B-NR a R b 、-B-OR a 、-BC(O)R a 、-BC(O)OR a 、-BC(O)NR a R awherein B is a bond, a C1-C3-alkyl group or a branched C1-C3-alkyl group; and R a and R b are each independently selected from the group consisting of hydrogen, C1-C3-alkyl, C3-C8-cycloalkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted cycloalkyl, substituted heterocyclyl, substituted aryl and substituted heteroaryl; and

[0024] n is an integer of 9-20, and preferably 12-20. In some preferred embodiments, R 1 is cyclopentyl, and R 2 Further, in some embodiments, n is preferably 12.

[0025] In some embodiments, the present method can take the form of a pharmaceutical composition comprising a compound as described herein or a pharmaceutically acceptable salt thereof as an active therapeutic agent and a pharmaceutically acceptable carrier. For example, in some embodiments, the composition can be a tablet having a core of 35% to 55% (by weight) of the active therapeutic agent and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can be, for example, microcrystalline cellulose, crospovidone type A, low-substituted hydroxypropyl cellulose, magnesium stearate, and colloidal anhydrous silicon dioxide.

[0026] The compounds and pharmaceutical compositions described herein have potency and selectivity for cancer stem cells, making them suitable for various anti-cancer therapeutic uses. For example, the present method can take the form of a method for preventing or reducing proliferation of at least one of cancer cells, cancer stem cells, and circulating tumor cells, wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0027] The present method may take the form of a method for treating cancer, wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0028] The present method may take the form of a method for treating or preventing metastatic disease, wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0029] The present method may take the form of a method for treating or preventing tumor recurrence, wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0030] The present method may take the form of a method for reducing resistance to cancer therapy (eg, chemotherapy resistance) wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0031] The present method may take the form of a method for treating or preventing at least one of radioresistance, chemotherapyresistance, and hormone therapy resistance, wherein a pharmaceutically effective amount of a compound or pharmaceutical composition as described herein is administered to a patient in need thereof.

[0032] It should be understood that one of ordinary skill in the art can determine the therapeutic dosage, dosage form and administration schedule of a particular embodiment using common methods known in the art.

[0033] The compounds of the present method can also be used to manufacture medicaments for various therapeutic uses, such as treating or preventing cancer, treating or preventing metastatic disease, and treating or preventing tumor recurrence.

[0034] Those of ordinary skill in the art will be able to recognize embodiments of the present method after reviewing the following detailed description.

[0035] Attached photos

[0036] Figure 1 Shown is a dose-response curve comparing compound [1C] with its parent compound using a mammosphere formation assay on the MCF7 cell line.

[0037] Figure 2 Dose-response curve comparing compound [2C] with its parent compound using mammosphere formation assay on MCF7 cell line.

[0038] Figure 3 Dose-response curve of the parent compound [3C] using the mammosphere formation assay on the MCF7 cell line.

[0039] Figure 4 Shown is a dose-response curve comparing compound [1C] with its parent compound using the Hoechst staining assay on the MCF7 cell line.

[0040] Figure 5 Shown is a dose-response curve comparing compound [2C] with its parent compound using the Hoechst staining assay on the MCF7 cell line.

[0041] Figure 6 Dose-response curves comparing compound [3C] to the parent compound using the Hoechst staining assay on the MCF7 cell line.

[0042] Figure 7 Shown is a dose-response curve comparing compound [1C] with its parent compound using the Hoechst staining assay on the hTERT-BJ1 cell line.

[0043] Figure 8Shown is a dose-response curve comparing compound [2C] with its parent compound using the Hoechst staining assay on the hTERT-BJ1 cell line.

[0044] Fig. 9 Dose-response curve of the parent compound [3C] using Hoechst staining assay in hTERT-BJ1 cell line.

[0045] describe

[0046] The following description includes currently contemplated modes of carrying out exemplary embodiments of the present method.The following description should not be taken in a limiting sense but is merely intended to illustrate the general principles of the invention.

[0047] According to the present method, compounds from three classes of CDK 4 / 6 inhibitors can be used as anticancer therapeutic agents. The first class includes substituted pyrrolopyrimidine compounds with fatty acid moieties. The second class includes substituted pyridopyrimidine compounds with fatty acid moieties. The third class includes substituted benzimidazole compounds with fatty acid moieties. The compounds described herein have useful drugs and medicinal properties. Many compounds exhibit significant selective CDK 4 / 6 inhibitory activity and are therefore valuable in treating a variety of clinical conditions in which CDK 4 / 6 kinases are abnormally elevated, activated, or present in normal amounts and activity, but all require inhibition of CDK to treat cell proliferative disorders. In particular, these compounds are promising as anticancer therapeutics. The following definitions describe compounds in each class, which are applicable to embodiments of the present method.

[0048] As used herein, the symbol C(O) represents a carbon-oxygen double bond. The term "halo" as used herein refers to a halogen, and includes fluorine, chlorine, bromine, or iodine, bonded as understood in the art.

[0049] The term "alkyl" as used herein refers to a saturated aliphatic group, including straight chain alkyl groups (e.g., methyl, ethyl, etc.), branched chain alkyl groups (isopropyl, tert-butyl, etc.), cycloalkyl (alicyclic) groups (cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkyl-substituted cycloalkyl, and cycloalkyl-substituted alkyl. The term "alkyl" also includes alkenyl and alkynyl. The general formula can be represented by the term "C n -alkyl", where n is an integer such as 1-20 to indicate a specific range or number of carbons in a particular alkyl group (straight or branched) in the group. For example, the term C1-C3-alkyl includes, but is not limited to, methyl, ethyl, propyl, and isopropyl. Similarly, the term C 3-6-Cycloalkyl includes but is not limited to cyclopropyl, cyclopentyl and cyclohexyl. Alkyl and cycloalkyl can be unsubstituted or substituted. Therefore, the term alkyl includes "unsubstituted alkyl" and "substituted alkyl", the latter refers to a portion having a substituent replacing a hydrogen on one or more carbons of the hydrocarbon backbone.

[0050] The term "alkenyl" includes unsaturated aliphatic analogs of a certain length that may replace the above alkyl groups but that contain at least one double bond. Alkenyl also includes "unsubstituted alkenyl" and "substituted alkenyl", the latter referring to moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone.

[0051] For example, the term "alkenyl" includes straight chain alkenyl (e.g., vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.), branched alkenyl, cycloalkenyl (alicyclic) group (cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl and cycloalkyl or cycloalkenyl substituted alkenyl. The term alkenyl also includes the alkenyl containing oxygen, nitrogen, sulfur or phosphorus atoms of one or more carbons of the replacement hydrocarbon backbone. In certain embodiments, straight or branched alkenyl has 6 or less carbon atoms in its main chain (e.g., straight chain is C2-C6, and branched chain is C3-C6). Similarly, cycloalkenyl can have 3-8 carbon atoms in its ring structure, and more preferably has 5 or 6 carbons in the ring structure. The term C2-C6 includes alkenyl containing 2 to 6 carbon atoms.

[0052] The term "alkynyl" includes unsaturated aliphatic analogs of certain lengths and possible substitutions for the above alkyl groups, but which contain at least one triple bond. In addition, the term alkynyl includes "unsubstituted alkynyls" and "substituted alkynyls," the latter referring to alkynyl moieties having substituents replacing hydrogens on one or more carbons of the hydrocarbon backbone.

[0053] As an example, the term "alkynyl" includes straight chain alkynyl (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.), branched chain alkynyl and cycloalkyl or cycloalkenyl substituted alkynyl. The term alkynyl also includes alkynyl containing oxygen, nitrogen, sulfur or phosphorus atoms of one or more carbons replacing the hydrocarbon backbone. In certain embodiments, straight or branched chain alkynyl has 6 or less carbon atoms in its backbone (e.g., straight chain is C2-C6, and branched chain is C3-C6). The term C2-C6 includes alkynyl containing 2 to 6 carbon atoms.

[0054] The term "substituted" is intended to describe a moiety having a substituent replacing a hydrogen on one or more atoms (e.g., C, O, or N) of the molecule. Such substituents may include, for example, but are not limited to, alkyl, alkoxy, alkenyl, alkynyl, halo, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonato, phosphinato, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino) groups. ), amido (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, thiol, alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonate, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, morpholino, phenol, benzyl, phenyl, piperazine, cyclopentane, cyclohexane, pyridine, 5H-tetrazole, triazole, piperidine, or aromatic or heteroaromatic moieties, and combinations thereof.

[0055] The term "amine" or "amino" shall refer to a molecule, or a moiety or a functional group as generally understood in the art, and may be a primary, secondary or tertiary amine. The term "amine" or "amino" includes compounds in which a nitrogen atom is covalently bonded to at least one carbon, hydrogen or heteroatom. The term includes, for example, but not limited to, "alkylamino", "arylamino", "diarylamino", "alkylarylamino", "alkylaminoaryl", "arylaminoalkyl", "alkylaminoalkyl", "amide", "amino" and "aminocarbonyl". The term "alkylamino" includes groups and compounds in which nitrogen is bonded to at least one additional alkyl group. The term "dialkylamino" includes groups in which a nitrogen atom is bonded to at least two additional alkyl groups. The terms "arylamino" and "diarylamino" include groups in which nitrogen is bonded to at least one or two aryl groups, respectively. The terms "alkylarylamino", "alkylaminoaryl" or "arylaminoalkyl" refer to an amino group bonded to at least one alkyl group and at least one aryl group. The term "alkylaminoalkyl" refers to an alkyl, alkenyl, or alkynyl group bonded to a nitrogen atom which is also bonded to an alkyl group.

[0056] The term "amide", "amino" or "aminocarbonyl" includes compounds or moieties containing a nitrogen atom that is bonded to the carbon of a carbonyl or thiocarbonyl group. The term includes "alkylaminocarbonyl" or "alkylaminocarbonyl", which includes alkyl, alkenyl, aryl or alkynyl groups bonded to an amino group that is bonded to a carbonyl group. It includes arylaminocarbonyl and arylcarbonylamino groups, which include aryl or heteroaryl moieties bonded to an amino group that is bonded to the carbon of a carbonyl or thiocarbonyl group. The terms "alkylaminocarbonyl", "alkenylaminocarbonyl", "alkynylaminocarbonyl", "arylaminocarbonyl", "alkylcarbonylamino", "alkenylcarbonylamino", "alkynylcarbonylamino" and "arylcarbonylamino" are included in the term "amide". Amides also include urea (aminocarbonylamino) and carbamate (carbamate) (oxycarbonylamino).

[0057] The term "aryl" includes a variety of groups, including 5- and 6-membered monocyclic aromatic groups that may include 0 to 4 heteroatoms, such as phenyl, pyrrole, furan, thiophene, thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole, oxazole, isoxazole, pyridine, pyrazine, pyridazine and pyrimidine, etc. In addition, the term "aryl" includes polycyclic aromatic groups, such as tricyclic, bicyclic, such as naphthalene, benzoxazole, benzodioxazole, benzothiazole, benzimidazole, benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline, anthracenyl, phenanthrenyl, naphthyridine, indole, benzofuran, purine, benzofuran, deazapurine or indolizine. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles", "heterocycles", "heteroaryls" or "heteroaromatics". The aromatic ring may be substituted at one or more ring positions with substituents such as alkyl, halogen, hydroxy, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonate, phosphine The term "alkyl" refers to any of a plurality of groups, for example, alkylamino, alkylamino, alkylaryl, alkylamino, alkylcarbonyl ...amino, alkylcarbonyl, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino, alkylamino,

[0058] As used herein, the term heteroaryl refers to a stable monocyclic or bicyclic ring having up to 7 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from O, N and S. Heteroaryl within the scope of this definition includes, but is not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline. As with the definition of heterocycle below, "heteroaryl" is also understood to include the N-oxide derivative of any nitrogen-containing heteroaryl. In the case where the heteroaryl substituent is bicyclic and one ring is non-aromatic or does not contain heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom-containing ring, respectively.

[0059] As used herein, the term "heterocycle" or "heterocyclyl" means a 5- to 10-membered aromatic or non-aromatic heterocyclic ring containing 1 to 4 heteroatoms selected from O, N and S, and includes bicyclic groups. Thus, "heterocyclyl" includes the above-mentioned heteroaryls and their dihydro and tetrahydro analogs. Additional examples of “heterocyclyl” include, but are not limited to, the following: benzimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, imidazolyl, indolyl, indolizinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolyl, tetrazolopyridinyl, thiadiazolyl, thiazolyl, thienyl, triazole

[0063] The compounds described herein include, but are not limited to, 1,4-dioxanyl, 1,4-dioxanyl, 1,6-dihydro-2-oxanyl, 1,4-dioxanyl, 1,6-dihydro-2-oxanyl, 1,4-dioxanyl, 1,6-dihydro-3-oxo-4-yl, 1,2-dioxin ... Attachment of a heterocyclyl substituent can be via a carbon atom or via a heteroatom.

[0060] The term "acyl" includes compounds and moieties containing an acyl group (CH3CO-) or a carbonyl group. The term "substituted acyl" includes acyl groups in which one or more hydrogen atoms are replaced by groups such as alkyl, alkynyl, halogen, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonate, phosphite, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), amido (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0061] The term "acylamino" includes moieties in which an acyl moiety is bonded to an amino group. For example, the term includes alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido.

[0062] The term "alkoxy" includes substituted and unsubstituted alkyl, alkenyl and alkynyl groups covalently attached to an oxygen atom. Examples of alkoxy groups include methoxy, ethoxy, isopropoxy, propoxy, butoxy and pentoxy, and may include cyclic groups such as cyclopentyloxy. Examples of substituted alkoxy groups include halogenated alkoxy groups. The alkoxy group can be substituted with groups such as alkenyl, alkynyl, halogen, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonate, phosphite, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), amido (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Examples of halogen-substituted alkoxy include, but are not limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, dichloromethoxy, trichloromethoxy, and the like.

[0063] The term "carbonyl" or "carboxyl" includes compounds and moieties containing carbon connected to an oxygen atom by a double bond, and tautomeric forms thereof. Examples of moieties containing carbonyls include aldehydes, ketones, carboxylic acids, amides, esters, anhydrides, etc. The term "carboxyl moiety" or "carbonyl moiety" refers to groups such as "alkylcarbonyl" in which an alkyl group is covalently bonded to a carbonyl, "alkenylcarbonyl" groups in which an alkenyl group is covalently bonded to a carbonyl, "alkynylcarbonyl" groups in which an alkynyl group is covalently bonded to a carbonyl, and "arylcarbonyl" groups in which an aryl group is covalently bonded to a carbonyl. In addition, the term also refers to groups in which one or more heteroatoms are covalently bonded to a carbonyl moiety. For example, the term includes moieties such as: aminocarbonyl moieties (in which a nitrogen atom is bonded to the carbon of a carbonyl group, such as amides), aminocarbonyloxy moieties in which oxygen and nitrogen atoms are bonded to the carbon of a carbonyl group (e.g., also referred to as "carbamates"). In addition, aminocarbonylamino (e.g., urea) also includes other combinations of carbonyls bonded to heteroatoms (e.g., nitrogen, oxygen, sulfur, etc. and carbon atoms). In addition, heteroatoms can be further substituted with one or more alkyl, alkenyl, alkynyl, aryl, aralkyl, acyl, etc.

[0064] The term "thiocarbonyl" or "thiocarboxyl" includes compounds and moieties containing a carbon connected to a sulfur atom by a double bond. The term "thiocarbonyl moiety" includes moieties similar to carbonyl moieties. For example, a "thiocarbonyl" moiety includes an aminothiocarbonyl group, wherein the amino group is bonded to the carbon atom of the thiocarbonyl group, and in addition, other thiocarbonyl moieties include oxythiocarbonyl (oxygen is bonded to the carbon atom), aminothiocarbonylamino, and the like.

[0065] The term "ether" includes compounds or moieties containing oxygen bonded to two different carbon atoms or heteroatoms. For example, the term includes "alkoxyalkyl," which refers to an alkyl, alkenyl, or alkynyl group covalently bonded to an oxygen atom that is also covalently bonded to another alkyl group.

[0066] The term "ester" includes compounds and moieties containing a carbon or heteroatom bonded to an oxygen atom which is also bonded to the carbon of a carbonyl group. The term "ester" includes alkoxycarboxyl groups such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, pentyloxycarbonyl, and the like. Alkyl, alkenyl, or alkynyl groups are as defined above.

[0067] The term "thioether" includes compounds and moieties containing a sulfur atom bonded to two different carbon or heteroatoms. Examples of thioethers include, but are not limited to, alkylthioalkyl, alkylthioalkenyl, and alkylthioalkynyl. The term "alkylthioalkyl" includes compounds having an alkyl, alkenyl, or alkynyl bonded to a sulfur atom bonded to an alkyl. Similarly, the terms "alkylthioalkenyl" and "alkylthioalkynyl" refer to compounds or moieties in which an alkyl, alkenyl, or alkynyl is bonded to a sulfur atom covalently bonded to an alkynyl.

[0068] The term "hydroxy" or "hydroxyl" includes groups having either -OH or -O-.

[0069] The term "polycyclyl" or "polycyclic radical" includes moieties having two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclyl) wherein two or more carbon atoms are common to two adjacent rings, e.g., the rings are "fused rings." Rings connected by non-adjacent atoms are referred to as "bridged" rings. Each ring of the polycyclic ring may be substituted with substituents as described above, for example, halogen, hydroxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxy, phosphate, phosphonate, phosphite, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino), amido (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0070] The term "heteroatom" includes atoms of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.

[0071] In addition, the phrase "any combination thereof" means that any number of the listed functional groups and molecules can be combined to create a larger molecular structure. For example, the terms "phenyl", "carbonyl" (or "=O"), "-O-", "-OH" and C 1-6 (ie, -CH3 and -CH2CH2CH2-) can combine to form a 3-methoxy-4-propoxybenzoic acid substituent. It should be understood that when combining functional groups and molecules to create larger molecular structures, hydrogens can be removed or added as needed to satisfy the valence state of each atom.

[0072] As will be appreciated by one of ordinary skill in the art, the compounds described herein include the bonds between adjacent atoms and / or hydrogens as required to satisfy the valence state of each atom. Bonds and / or hydrogen atoms are added, if necessary, to provide the following total number of bonds to each of the following types of atoms: carbon: four bonds; nitrogen: three bonds; oxygen: two bonds; and sulfur: two to six bonds.

[0073] The term "salt" of a compound relates to the corresponding salt prepared by using an acid selected from inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid, and organic acids such as tartaric acid, acetic acid, trifluoroacetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid and succinic acid, and alkylsulfonic acids such as methanesulfonic acid, ethanesulfonic acid, ethane-1,2-disulfonic acid and 2-hydroxyethanesulfonic acid, and arylsulfonic acids such as benzenesulfonic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid and naphthalene-1,5-disulfonic acid.

[0074] As used herein, the phrase "pharmaceutically effective amount" means the amount necessary to administer to a host or to a cell, tissue or organ of a host to achieve a therapeutic result (e.g., to regulate, modulate or inhibit protein kinase activity, such as inhibiting protein kinase activity, or treating cancer). A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian can start with a dose of a compound of the present invention in a pharmaceutical composition that is lower than the level required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved.

[0075] The term "about" means that a value falls within the generally accepted standard range of average error when considered by one of ordinary skill in the art. As expected, the meaning of "about" depends on the context in which it is used. Generally, the term "about" may refer to ±5%, preferably ±2.5%, and more preferably ±1% of the value or range to which it refers. For example, in the context of weight percentages, the phrase "about 20%" may mean 20% ±5%, preferably 20% ±2.5%, and more preferably 20% ±1%.

[0076] The terms "treat", "treated", "treating" and "treatment" include reducing or alleviating at least one symptom associated with or caused by the condition, disorder or disease being treated, particularly cancer. In certain embodiments, treatment includes reducing and / or alleviating at least one symptom associated with or caused by the cancer being treated by the compounds of the present invention. For example, treatment can be the reduction of one or several symptoms of cancer, or the complete eradication of cancer.

[0077] The compounds described herein include a fatty acid moiety as described in the present disclosure. As used herein, a fatty acid is a carboxylic acid having an aliphatic chain, which may be saturated or unsaturated, but saturated chains are preferred. Examples of saturated fatty acids include lauric acid (CH3(CH2) 10 COOH), palmitic acid (CH3(CH2) 14 COOH), stearic acid (CH3(CH2) 16 COOH) and myristic acid (CH3(CH2) 12COOH). Oleic acid (CH3(CH2)7CH=CH(CH2)7COOH) is an example of a naturally occurring unsaturated fatty acid. Reference may also be made to salts or esters of fatty acids and to fatty amide moieties thereof, but for simplicity these are included in the meaning of fatty acid moieties as used herein. For example, myristic acid may be referred to as myristate, and oleic acid may be referred to as oleate. The fatty acid moiety may also be the carbonyl group of a fatty acid, i.e., the group formed by the loss of a hydroxyl group of a carboxylic acid. In some embodiments, the fatty acid moiety may be bonded to the therapeutic agent via an amide bond. For example, a myristic acid conjugate may have the fatty acid moiety CH3(CH2) 12 CO-NH-, where the tertiary nitrogen is bonded to the therapeutic agent: and n is an integer from 1 to 20, and preferably from 10 to 20. When the myristate moiety is conjugated by myristoylation, this may form a myristamide (or myristic amide) group.

[0078] Substituted pyrrolopyrimidine compounds

[0079] In some embodiments of the present method, the first class of anticancer CDK 4 / 6 inhibitors are substituted pyrrolopyrimidine compounds and pharmaceutically acceptable salts thereof. It should be understood that some compounds in the first class are derivatives of the parent compound 7-cyclopentyl-N,N-dimethyl-2-[(5-piperazin-1-ylpyridin-2-yl)amino]pyrrolo[2,3-d]pyrimidine-6-carboxamide (also known as ribociclib). Some embodiments in the first class have a chemical structure shown in the following general formula [1A], wherein the fatty acid portion is bonded to piperazine.

[0080]

[0081] As used in the general formula [1A]:

[0082] R 4 is selected from the group consisting of hydrogen, C1-C8-alkyl, substituted C1-C8-alkyl, C3-C8-cycloalkyl, substituted C3-C8-cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl;

[0083] Z is CR z , where R z is selected from halogen, hydrogen, C1-C3-alkyl, C1-C3-alkoxy, CN, C═NOH, C═NOCH3, C(O)H, C(O)C1-C3-alkyl, C3-C8-cycloalkyl, heterocyclyl, aryl, heteroaryl, substituted C1-C3-alkyl, substituted C3-C8-cycloalkyl, substituted heterocyclyl, substituted aryl, substituted heteroaryl, -B-NR a R b 、-B-OR a 、-BC(O)Ra 、-BC(O)OR a 、-BC(O)NR a R a ; wherein B is a bond, C1-C3-alkyl or branched C1-C3-alkyl; and R a and R b are each independently selected from hydrogen, C1-C3-alkyl, C3-C8-cycloalkyl, heterocyclyl, aryl, heteroaryl, substituted alkyl, substituted cycloalkyl, substituted heterocyclyl, substituted aryl and substituted heteroaryl; and

[0084] In the fatty acid moiety, "n" represents an integer of 9 to 20, and preferably 12 to 20.

[0085] It will be appreciated that pharmaceutically acceptable salts may also be used. As described above, the salts may be prepared using, for example, an acid selected from inorganic acids, organic acids, alkylsulfonic acids, ethanesulfonic acids, and arylsulfonic acids. In some preferred embodiments of the present method, the first class of anticancer CDK 4 / 6 inhibitors is a compound having the general formula [1B] as shown below. In these embodiments, R 4 is C5-cycloalkyl, Z is dimethylformamide or acetyl, and n represents an integer from 9 to 20 and more preferably from 12 to 20. The parent compound of this general formula is ribociclib, an FDA-approved drug for the treatment of HR-positive, HER2-negative advanced or metastatic breast cancer (with an aromatase inhibitor). However, embodiments of the present method according to formula 1[A] have a C5-cycloalkyl conjugated at the terminal piperazine. 11 -C 22 Fatty acid moiety. Preferably, the fatty acid moiety is straight chain and saturated. In some preferred embodiments, the fatty acid moiety is one of lauric acid, myristic acid, palmitic acid and stearic acid. The fatty acid moiety significantly improves the cellular uptake of the compound, greatly enhancing its inhibitory effect on cancer stem cell proliferation and selectivity for tumor cells.

[0086]

[0087] The following exemplary embodiment shows compound [1C], wherein R 4 is an unsubstituted C5-cycloalkyl group, Z is dimethylformamide, and n is 12. As a result, this embodiment has a 14-carbon fatty acid (i.e., myristate) moiety. The compound shown as compound [1C] has been synthesized, and in a mammosphere assay, the inhibitory effect of the compound on MCF7 cells was significantly enhanced compared to the known anticancer therapeutic drug ribociclib (at a concentration of 1 μM to 100 μM), indicating that the fatty acid moiety has an incredible effect on the compound. In preliminary laboratory evaluations, this embodiment effectively inhibited 100% of cell proliferation at concentrations as low as 1 μM, showing excellent anticancer efficacy. For example, Figure 1 Dose-response curves for compound [1C] and its parent compound are shown and demonstrate improved CSC inhibition due to the addition of the fatty acid moiety. It should be understood that similar results are expected for other fatty acid moieties as few as 11 carbons and as many as 22 carbons.

[0088]

[0089] In some embodiments of the present method, the first class of anticancer CDK 4 / 6 inhibitors is a substituted pyrrolopyrimidine compound as shown in the following general formula [1D]. Compared with general formula [1A], the compound having general formula [1D] includes a fatty acid moiety at Z. It should be understood that pharmaceutically acceptable salts may also be used.

[0090]

[0091] As used in the general formula [1D]:

[0092] R 4 is selected from the group consisting of hydrogen, C1-C8-alkyl, substituted C1-C8-alkyl, C3-C8-cycloalkyl, substituted C3-C8-cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl;

[0093] R 3 is selected from hydrogen, OH, C1-C8-alkyl, substituted C1-C8-alkyl, C3-C8-cycloalkyl, C(O)C1-C8-alkyl, C1-C8-haloalkyl, C1-C8-cyanoalkyl, C1-C8-alkyl-OH, SO2-C1-C8-alkyl, C1-C8-alkyl-C3-C8-cycloalkyl and C1-C8-alkoxy, when R 3 When other than hydrogen, it may be substituted or unsubstituted; and

[0094] In the fatty acid moiety, "n" represents an integer of 9 to 20, and preferably 12 to 20.

[0095] In preferred embodiments of the present method, the first class of anticancer CDK 4 / 6 inhibitors is a compound having the general formula [1E] as shown below. In these embodiments, R 4 is C5-cycloalkyl, R 3 is hydrogen, and n represents an integer of 9 to 20, and more preferably 10-20, and more preferably 10-16. The compound having the general formula [1E] is a derivative of ribociclib in which a fatty acid moiety replaces the dimethyl-amino group at the carboxyl group.

[0096]

[0097] The following exemplary embodiments show compounds of formula [1F], wherein R4 is an unsubstituted C5-cycloalkyl group, R 3 is hydrogen, and n is 12. As a result, this embodiment has a 14-carbon fatty acid (ie, myristate) moiety. Compound [1F] is expected to show improved inhibitory effects in the mammosphere assay.

[0098]

[0099] Table 1 below summarizes the results of multiple assays comparing compound [1C] and its parent compound (ribociclib) and contains in vitro biological data for both compounds. When compared to the parent compound, compound [1C] showed a 7-fold increase in potency in the 3D mammosphere assay (IC 50 0.2 vs. 1.5 μM), while maintaining similar activity in a 2D cell viability assay (IC 50 Both were about 2 μM). This shows that conjugation with the fatty acid moiety not only significantly improved the potency in the 3D mammosphere assay. Comparison of the mammosphere monolayer selectivity index ("SI") showed that conjugation also improved the selectivity for mammospheres (SI 10 vs. 1.3). Both compounds were non-toxic to the non-tumor cell line hTERT-BJ1 at concentrations up to 90 μM, representing high selectivity for tumor cell lines. It can be seen that the compounds according to the present method result in a significant increase in potency and selectivity for tumor cells.

[0100]

[0101] Table 1. Mammosphere assay results for exemplary compounds in the first category.

[0102] As used in the table, “IC 50 The term “IC 50 The term “IC 50 The term “SI (mammosphere / monolayer MCF7)” refers to the half maximal inhibitory concentration in a 2D cell viability assay using the immortalized non-tumor fibroblast cell line hTERT-BJ1. The term “SI (mammosphere / monolayer MCF7)” refers to the mammosphere selectivity index, which is the IC obtained by comparing bioactivity against MCF7 in 3D and 2D assays. 50 The term “SI (BJ1 / MCF7 monolayer)” refers to the cancer selectivity index, which is the IC obtained by comparing the bioactivity of MCF7 and hTERT-BJ1 in a cell viability assay. 50 The ratio between the values.

[0103] Substituted pyridopyrimidine compounds

[0104] The second class comprises substituted pyridopyrimidines having a fatty acid moiety, as shown in the general formula [2A] below. It should be understood that some embodiments in the second class comprise derivatives of 6-acetyl-8-cyclopentyl-5-methyl-2-(5-piperazine-1-yl-pyridin-2-ylamino)-8H-pyrido[2,3-d]pyrimidin-7-one (also known as Palbociclib). It should also be understood that pharmaceutically acceptable salts, such as those mentioned above, may also be used, as understood by those skilled in the art. Other exemplary salts include malate, tartrate, bromide, hydrogen bromide dihydrate, hydrogen chloride, sulfate dihydrate, camphorsulfonate, naphthalenesulfonate, naphthalenesulfonate dihydrate, toluenesulfonate, citrate monohydrate, maleate and oxalate.

[0105]

[0106] In general formula 2[A]:

[0107] R 1 is hydrogen, aryl, C1-C8 alkyl, C1-C8 alkoxy, C3-C7 cycloalkyl or C3-C7 heterocyclyl;

[0108] R 2 independently selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 acyl, C3-C7 cycloalkyl, C1-C8 alkoxy, C1-C8 alkoxyalkyl, C1-C8 haloalkyl, C1-C8 hydroxyalkyl, C2-C8 alkenyl, C2-C8 alkynyl, nitrile, nitro, OR 5 , SR 5 NR 5 R 6 、N(O)R 5 R 6 、P(O)(OR 5 )(OR 6 ), (CR 5 R 6 ) m NR 7 R 8 , COR 5 , (CR 4 R 5 ) m C(O)R 7 、CO2R 5 ,CONR 5 R 6 、C(O)NR 5 S02R 6 NR 5S02R 6 、C(O)NR 5 OR 6 、S(O) n R 5 、SO2NR 5 R 6 、P(O)(OR 5 )(OR 6 ), (CR 5 R 6 ) m P(O)(OR 7 )(OR 8 ), (CR 5 R 6 ) m -aryl, (CR 5 R 6 ) m -heteroaryl and -CR 5 ═CR 6 C(O)R 7 ;

[0109] In each instance, R 3 are independently hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl or C3-C7 cycloalkyl;

[0110] R 5 , R 6 , R 7 and R 8 are independently hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, aralkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl or heteroarylalkyl; m is 0 to 6; and

[0111] In the fatty acid moiety, n represents an integer of 9 to 20, and preferably 12 to 20.

[0112] The general formula [2B] shown below is a general formula of a preferred embodiment of the second class of the present method. In the general formula [2B], R 1 is an unsubstituted C5-cycloalkyl group, R 2 is C1-acyl (acetyl), R 3 is a methyl group, and n represents an integer of 9 to 20 and more preferably 12 to 20.

[0113]

[0114] The second exemplary embodiment is shown in the following compound [2C], wherein R 1 is an unsubstituted C6-cycloalkyl group, R 2 is C1-acyl (acetyl), R 3is methyl, and n is 12. As a result, this embodiment has a 14-carbon fatty acid (i.e., myristate) moiety. Compound [2C] has been synthesized, and in a mammosphere assay, the inhibitory effect of the compound on MCF7 cells was significantly enhanced compared to the known anticancer therapeutic drug palbociclib (at concentrations of 1 μM to 100 μM). These results also indicate that for the second class of compounds according to the present method, the fatty acid moiety has a significant beneficial effect on the anticancer efficacy of the compound. For example, Figure 2 Dose-response curves for compound [2C] and its parent compound are shown and demonstrate improved CSC inhibition due to the addition of the fatty acid moiety. It will be appreciated that similar results are expected for other fatty acid moieties as few as 11 carbons and as many as 22 carbons.

[0115]

[0116] The second class also includes substituted pyridopyrimidines having a fatty acid moiety as shown in the following general formula [2D]. As can be seen, the fatty acid moiety in formula [2D] is conjugated at the pyrido[2,3-d]pyrimidine, as opposed to the piperazine seen in formula [2A].

[0117]

[0118] In general formula 2[D]:

[0119] R 1 is hydrogen, aryl, C1-C8 alkyl, C1-C8 alkoxy, C3-C7 cycloalkyl or C3-C7 heterocyclyl;

[0120] R 2 is selected from hydrogen, halogen, C1-C8 alkyl, C1-C8 acyl, C3-C7 cycloalkyl, C1-C8 alkoxy, C1-C8 alkoxyalkyl, C1-C8 haloalkyl, C1-C8 hydroxyalkyl, C2-C8 alkenyl, C2-C8 alkynyl, nitrile, nitro, OR 5 , SR 5 NR 5 R 6 、N(O)R 5 R 6 、P(O)(OR 5 )(OR 6 ), (CR 5 R 6 ) m NR 7 R 8 , COR 5 , (CR 4 R 5 ) m C(O)R 7 、CO2R5 ,CONR 5 R 6 、C(O)NR 5 S02R 6 NR 5 S02R 6 、C(O)NR 5 OR 6 、S(O) n R 5 、SO2NR 5 R 6 、P(O)(OR 5 )(OR 6 ), (CR 5 R 6 ) m P(O)(OR 7 )(OR 8 ), (CR 5 R 6 ) m -aryl, (CR 5 R 6 ) m Heteroaryl and -CR 5 ═CR 6 C(O)R 7 ;

[0121] In each instance, R 3 are independently hydrogen, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl or C3-C7 cycloalkyl;

[0122] R 5 , R 6 , R 7 and R 8 is independently selected from hydrogen, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, aralkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl or heteroarylalkyl; m is 0 to 6; and

[0123] In the fatty acid moiety, "n" represents an integer of 9 to 20, and preferably 12 to 20.

[0124] The general formula [2E] shown below is an example of another preferred embodiment of the second class according to the present method. In the general formula [2E], R 1 is an unsubstituted C5-cycloalkyl group, R 2 For H, R 3 is a methyl group, and n represents an integer of 9 to 20 and more preferably 12 to 20.

[0125]

[0126] The second exemplary embodiment is shown in the following compound [2F], wherein R 1 is an unsubstituted C5-cycloalkyl group (e.g., cyclopentyl), R 2 For H, R 3 is methyl, and n is 12. As a result, this embodiment has a 14-carbon fatty acid (ie, myristate) moiety.

[0127]

[0128] Table 2 below summarizes the results of multiple tests comparing compound [2C] and its parent compound (palbociclib) and includes in vitro biological data for both compounds. The results show that compound [2C] has reduced potency in the 3D mammosphere assay (IC 50 =5.1 vs. about 0.2 μM). However, compound [2C] was nontoxic in the 2D cell viability assay at concentrations up to 30 μM, whereas palbociclib showed an IC 50 is about 0.1 μM. In addition, when comparing 2D and 3DMCF7 assays, palbociclib is non-selective. These results indicate that compound [2C] exhibits improved compound selectivity for 3D mammospheres, as opposed to normal cells. Therefore, it should be understood that the compounds of the present method can be used to selectively target cancer cells, particularly CSCs. Both compounds were non-toxic to the non-tumor cell line hTERT-BJ1 at concentrations up to 90 μM, representing high selectivity for tumor cell lines. Therefore, the compounds in the second class of the present method exhibit higher selectivity in targeting cancer stem cells.

[0129]

[0130] Table 2. Mammosphere assay results for exemplary compounds in the second category.

[0131] Substituted benzimidazole compounds

[0132] The third class includes substituted benzimidazole compounds having the general formula [3A] as shown below. It should also be understood that pharmaceutically acceptable salts, such as those mentioned above, may also be used, as understood by a person skilled in the art. It should be understood that some embodiments in the third class include derivatives of N-[5-[(4-ethylpiperazine-1-yl)methyl]pyridin-2-yl]-5-fluoro-4-(7-fluoro-2-methyl-3-propane-2-ylbenzimidazol-5-yl)pyrimidine-2-amine, also known as Abemaciclib. Like the compounds in the first two classes, the compounds in the third class are also potent CDK 4 / 6 inhibitors.

[0133]

[0134] In Formula 3A,

[0135] R 1 Fatty acid part wherein m is an integer from 0 to 4 and more preferably from 0 to 2, such that when m is 0, there is a direct bond to the nitrogen in piperazine, and n is an integer from 9 to 20 and more preferably from 12 to 20;

[0136] R 2 is H or C1-C3 alkyl;

[0137] R 3 and R 4 is H or fluorine, and R 3 and R 4 At least one of them is fluorine;

[0138] R 5 is C3-C5 alkyl, C3-C5 cycloalkyl or cyclopropyl-methyl;

[0139] R 6 is H or C1-C3 alkyl; and

[0140] X is a bond, a C1-C3 alkyl group, O, or S.

[0141] The general formula [3B] shown below is a general formula of a preferred embodiment of the third class of the present method. In the general formula [3B], R 1 Fatty acid part wherein m is an integer from 0 to 4 and more preferably from 0 to 2, such that when m is 0, there is a direct bond to the nitrogen in piperazine, and n is an integer from 9 to 20 and more preferably from 12 to 20; R 2 For H, R 3 and R 4 is fluorine, R 5 is C3 alkyl (isobutyl), and R6 is methyl.

[0142]

[0143] Compound [3C] shown below is a formula of an exemplary preferred embodiment of the third class according to the present method. In the general compound [3C], R 1 The fatty acid part Where m is 0 and n is 12, R 2 For H, R 3 and R 4 is fluorine, R 5 is C3 alkyl (isobutyl), and R 6is methyl. Embodiments wherein n is 9 to 20 are contemplated and planned for evaluation. The embodiment shown as compound [3C] is a derivative of abemaciclib, a CDK 4 / 6 inhibitory compound approved by the FDA for the treatment of advanced and metastatic breast cancer. As described herein, compound [3C] and other compounds having formula [3A] are expected to effectively inhibit CDK 4 / 6, making them particularly suitable for use as anticancer therapeutics and selectively targeting and inhibiting cancer cells and CSCs.

[0144]

[0145] A second exemplary embodiment of the third class of compounds is shown below in Formula 3D. In this example, R 1 The fatty acid part Where m is 0 and n is 12, R 2 For H, R 3 and R 4 is fluorine, R 5 is C3 alkyl (isobutyl), and R 6 As described herein, like compound [3C], this embodiment is expected to be more potent than abemaciclib in terms of CDK 4 / 6 inhibition and selectivity, and is particularly suitable for use as an anticancer therapeutic agent.

[0146]

[0147] Table 3 below shows the in vitro biological data of abemaciclib. It can be seen that the compound has high efficacy (IC 50 <0.04 μM). Compounds of formula [3A] comprising various fatty acid moieties are being evaluated. As can be seen from Table 3, abemaciclib is highly specific and highly effective against CSCs. Therefore, abemaciclib can be used as a therapeutic agent for specifically targeting CSCs. Due to its selectivity, abemaciclib can be used to treat and / or prevent tumor recurrence and / or metastasis, as well as for targeting circulating tumor cells.

[0148]

[0149] Table 3. Mammosphere trial results of abemaciclib.

[0150] Figure 1-3 It is the dose-response curve of various compounds described herein obtained using the mammosphere assay. The assay was performed using the MCF7 cell line. The curve is plotted as the mean of two independent experiments, and the standard deviation of each point is represented by a vertical line. The inhibition percentage is shown as a function of the logarithm of the concentration (μM). Figure 1 Shown is a dose-response curve comparing compound [1C] with its parent compound (ribociclib) using a mammosphere formation assay on the MCF7 cell line. Figure 4 The dose-response curves comparing compound [1C] and its parent compound using the Hoechst staining assay on the MCF7 cell line are shown. It can be seen that compound [1C] is more effective against MCF7 cells at each concentration. This suggests that compounds of formula [1A], particularly compounds of formula [1B], have efficacy as effective anticancer therapeutic agents. Figure 7 A dose-response curve comparing compound [1C] to its parent compound using a Hoechst staining assay on the hTERT-BJ1 cell line is shown. The parent compound was more potent at most concentrations tested, indicating that compounds of general formula [1A] and particularly general formula [1B] are more selective for CSCs. Similar effects are expected for compounds of general formula [1D].

[0151] In addition, although the data disclosed herein relate to MCF7 and hTERT-BJ1 cell lines, the compounds of the present method are effective for other types of cancer. In previous work, the inventors demonstrated that mitochondrial biogenesis inhibitors successfully inhibited the formation of tumor spheres in a variety of tumor type cell lines. Table 4 below lists cancer cell lines that are sensitive to mitochondrial biogenesis inhibitors. In view of these results, the present method is effective for many cancer types.

[0152]

[0153] Table 4. Mitochondrial biogenesis inhibitors are effective against various cancer types.

[0154] The present method describes a pharmaceutical composition comprising a therapeutically effective amount of a compound from the first or second class or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent or excipient thereof. The compound according to the present method can be used as an anticancer therapeutic agent. The compound can be administered to a subject in a pharmaceutically effective amount in a pharmaceutically acceptable carrier according to methods known in the art. In some embodiments, the compound of the present method can be used in combination with other cancer therapies, such as, but not limited to, chemotherapeutic agents, mitochondrial biogenesis inhibitors (e.g., mitoriboscins, mitoketoscins, repurposcins such as antimitoscins), radiotherapy, phototherapy, and caloric restriction.

[0155] It should be understood that those skilled in the art can use common and known methods in the art to develop formulations for specific embodiments. In some embodiments, the pharmaceutical composition may be in the form of a tablet, capsule or pill. The pharmaceutical composition may have a dosage of 20 mg to 500 mg of the therapeutic composition. In some embodiments, the pharmaceutical composition may include a tablet with 200 mg of a therapeutic compound (e.g., the above-mentioned compound, e.g., compound [1C]). The tablet may contain at least about 35%, 40%, 45%, 50% or 55% of the therapeutic compound content, measured as a w / w percentage of the therapeutic compound (as a free base) of the tablet core.

[0156] The tablet may have a core formed of microcrystalline cellulose, crospovidone type A, low-substituted hydroxypropyl cellulose, magnesium stearate, and colloidal anhydrous silicon dioxide. In a first exemplary embodiment, a tablet having 200 mg of a therapeutic compound (e.g., compound [1C]) may include an inner core having microcrystalline cellulose (67.44 mg), hydroxypropyl cellulose (48.12 mg), crospovidone (29.20 mg), colloidal silicon dioxide (anhydrous) (2.12 mg), and magnesium stearate (6.36 mg), and an outer core having crospovidone (12.84 mg), colloidal silicon dioxide (anhydrous) (1.06 mg), and magnesium stearate (8.46 mg). In a second exemplary embodiment, the tablet may have about 10% to about 45% (w / w) of the therapeutic compound (e.g., compound [2C]), and preferably about 18% to about 28% of the therapeutic compound; about 4% to about 18% of a water-soluble acid; about 20% to about 75% of a diluent; about 5% to about 18% of a disintegrant; about 0.2% to about 10% of a lubricant; and, optionally, about 0% to about 5% of a glidant and about 0% to about 15% of a binder. It should be understood that the pharmaceutical compositions of the present method can be very similar to pharmaceutical compositions comprising the parent compound. For example, International Patent Application Publication No. WO2016 / 166703, filed April 14, 2016, describes an example of a ribociclib tablet formulation, and the entire contents of which are incorporated by reference. As another example, International Patent Application Publication No. WO 2016 / 193860, filed on May 24, 2016, describes solid dosage forms of palbociclib, and is incorporated by reference in its entirety.

[0157] The tablets may have a film coating. The film coating may include black iron oxide, red iron oxide, soy lecithin, polyvinyl alcohol (partially hydrolyzed), talc, titanium dioxide, and xanthan gum. Depending on the desired appearance of the final tablet, the tablets may be coated with commercially available coating premixes. For example, (Colorcon, Harleysville, PA) has the appearance of a HPMC (hydroxypropyl methylcellulose) coating material and has the following composition: HPMC (Pharmacoat 603) 71.4%, polyethylene glycol 7.15%, talc 7.15% and iron oxide 14.3%.

[0158] Selective inhibition of CDK 4 / 6 also suggests that the compounds described herein may be used to reduce or eliminate drug resistance and / or treatment resistance in cancer. Due to their inhibitory activity against CDK and other kinases, the compounds of the present method are also useful research tools for studying the mechanism of action of such kinases and can be used in vitro and in vivo.

[0159] The treatment methods described herein are preferably carried out by administering a therapeutically effective amount of the first or second class compound to a subject in need of treatment. These compounds are easily synthesized using the reaction steps described below, or alternative reaction steps can be used. Those skilled in the art can easily find alternative reaction steps after consulting the present invention, and the alternative reaction steps include "Comprehensive Organic Synthesis", Trost, Fleming, Pergamon: 1991 and "Comprehensive organic Functional group Transformations", Katritky, Meth-Cohn, Rees, Pergamon: 1995. The reaction steps described can be administered by a variety of routes, including oral and parenteral, and have little or no toxicity.

[0160] The following abbreviations may be used in the following discussion of exemplary synthetic methods: N-methylmorpholine (NMM), dichloromethane (DCM), dimethylformamide (DMF), ethyl acetate (EtOAc), sodium bicarbonate (NaHCO3), sodium sulfate (Na2SO4), methanol (MeOH). In the following description, [M+H] + refers to the protonated molecule and the value indicated is the mass of the protonated molecule. RT refers to the solute retention time.

[0161] Analytical LC-MS: Waters Sunfire C18 30×4.6 mm column, gradient eluent 3-97% acetonitrile / water with 0.05% formic acid. Time 0-6 minutes. Preparative HPLC: LC column: Phenomenex Kinetex 5μm EVO C18 100 250×21.2 mm. Gradient eluent: 40-95% acetonitrile / water with 0.1% formic acid.

[0162] In the first synthetic example, compound [2C], also known as 6-acetyl-8-cyclopentyl-5-methyl-2-[[5-(4-tetradecanoylpiperazin-1-yl)-2-pyridinyl]amino]pyrido[2,3-d]pyrimidin-7-one, as shown above, was synthesized using palbociclib (obtained from LC laboratories, Woburn, MA, USA). Tetradecanoic acid (0.104 g, 0.46 mmol) was dissolved in thionyl chloride at room temperature. The solution was refluxed for 60 minutes, concentrated under reduced pressure, and the residue was dissolved in anhydrous DCM (2 ml) at room temperature to give a 0.227 M acid chloride stock solution. The acid chloride stock solution (0.25 ml, 0.057 mmol) was added to a stirred mixture of 6-acetyl-8-cyclopentyl-5-methyl-2-[[5-(1-piperazinyl)-2-pyridinyl]amino]-pyrido[2,3-d]pyrimidin-7(8H)-one (0.024 g, 0.05 mmol) and NMM (18 μl, 0.16 mmol) in DCM (1 ml) and DMF (0.5 ml). The mixture was stirred at room temperature for 90 minutes. The solvent was evaporated under reduced pressure and the residue was dissolved in EtOAc (30 ml), washed with saturated NaHCO (15 ml) and brine (15 ml), and then dried over NaSO. The drying agent was separated by filtration, and the filtrate was concentrated under reduced pressure to produce a crude product. The crude product was triturated with diethyl ether and the resulting light brown solid was collected by filtration, washed with diethyl ether and dried under vacuum to give 6-acetyl-8-cyclopentyl-5-methyl-2-[[5-(4-tetradecanoylpiperazin-1-yl)-2-pyridinyl]amino]pyrido[2,3-d]pyrimidin-7-one (0.0145 g). LC-MS 658.2 [M+H] + , RT 4.12 min.

[0163] In the second synthetic example, the above-mentioned compound [1C], also known as 7-cyclopentyl-N,N-dimethyl-2-[[5-(4-tetradecanoylpiperazin-1-yl)-2-pyridinyl]amino]pyrrolo[2,3-d]pyrimidine-6-carboxamide, was synthesized using the same method as in the first synthetic example, except that ribociclib (obtained from LC laboratories, Woburn, MA, USA) (0.022 g, 0.05 mmol) was reacted with the acid chloride stock solution instead of palbociclib. The crude product was purified on silica gel (2-4% MeOH / DCM) to give 7-cyclopentyl-N,N-dimethyl-2-[[5-(4-tetradecanoylpiperazin-1-yl)-2-pyridinyl]amino]pyrrolo[2,3-d]pyrimidine-6-carboxamide (0.0166 g) as a light brown solid. LC-MS 645.2 [M+H] +,RT 2.72 minutes.

[0164] In the third synthesis example, the compound [3C] shown above is synthesized by a series of intermediate compounds as shown below, also known as 1-[4-[[6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzimidazole-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]methyl]piperazine-1-yl]tetradecane-1-one. First, the intermediate compound [4A] as shown below and referred to as 4-tetradecanoylpiperazine-1-carboxylic acid tert-butyl ester is synthesized as follows. Under a nitrogen atmosphere, isobutyl chloroformate (0.65ml, 5.0mmol) is added to a stirred solution of tetradecanoic acid (1.26g, 5.5mmol) and NMM (0.73ml, 5.5mmol) in anhydrous DCM (20ml) at room temperature. After 4 hours, an anhydrous DCM (5ml) solution of 1-Boc-piperazine (0.93g, 5.0mmol) is added to the mixture. The mixture is stirred for 16 hours. The solvent was removed under reduced pressure to give a crude product. The crude product was dissolved in EtOAc (75 ml), washed with 2M HCl (50 ml), saturated NaHCO3 (40 ml) and brine (30 ml), and then dried over MgSO4. After filtration, the solvent was evaporated under reduced pressure to give tert-butyl 4-tetradecanoylpiperazine-1-carboxylate (1.76 g) as a white solid. LC-MS 397.2 [M+H] + ,RT 4.02 minutes.

[0165]

[0166] Second, the intermediate compound [4B], as shown below and referred to as 1-piperazine-1-yltetradecane-1-one, was synthesized as follows. A solution of 4-tetradecanoylpiperazine-1-carboxylic acid tert-butyl ester (0.51 g, 1.26 mmol) in a 1:1 mixture of anhydrous DCM (10 ml) and trifluoroacetic acid or TFA (10 ml) was stirred at room temperature for 90 minutes under a nitrogen atmosphere, and the solvent was removed under reduced pressure to give a crude product. The crude product was dissolved in EtOAc (30 ml), washed with saturated NaHCO3 (15 ml) and brine (15 ml), and dried over MgSO4. After filtration, the solvent was evaporated under reduced pressure to give 1-piperazine-1-yltetradecane-1-one (0.33 g) as a white waxy solid. LC-MS 297.3 [M+H] + ,RT 1.81 minutes.

[0167]

[0168] Third, intermediate compound [4C] shown below and designated as 6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl]amino]pyridine-3-carbaldehyde was prepared as follows. A suspension of 6-aminopyridine-3-carbaldehyde (0.076 g, 0.625 mmol), 6-(2-chloro-5-fluoro-pyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methylbenzimidazole (0.161 g, 0.500 mmol), Xantphos (0.0276 g, 0.0476 mmol), palladium chloride (0.0056 g, 0.0315 mmol) and K2CO3 (0.069 g, 0.500 mmol) in 2-methyl-2-butanol (4 ml) was heated at +100°C for 18 hours in a sealed tube. The reaction mixture was cooled to room temperature and diluted with EtOAc (30 ml) and water (30 ml). The precipitate with the aqueous phase was separated and collected by filtration. The light brown solid was washed with water (30 ml) and acetone (20 ml) and dried under vacuum to give 6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl]amino]pyridine-3-carboxaldehyde (0.125 g) as a light brown solid. LC-MS 409.0[M+H] + ,RT 2.07 minutes.

[0169]

[0170] Fourth, intermediate compound [4D] shown below and referred to as 1-[4-[[6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl]amino]-3-pyridinyl]-methyl]piperazin-1-yl]tetradecan-1-one, also shown above as compound [3C] was synthesized as follows. A suspension of 6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl]amino]pyridine-3-carbaldehyde (0.050 g, 0.122 mmol), 1-piperazin-1-yltetradecan-1-one (0.030 g, 0.100 mmol) and sodium triacetoxyborohydride (0.212 g, 1.00 mmol) in DCE (20 ml) was heated at +60° C. for 90 minutes in a sealed tube. The reaction mixture was cooled to room temperature, diluted with DCM (30 ml), washed with water (10 ml) and brine (10 ml), and dried over MgSO4. After filtration, the solvent was evaporated under reduced pressure to give a crude product (0.1564 g), which was purified by preparative HPLC to give 1-[4-[[6-[[5-fluoro-4-(7-fluoro-3-isopropyl-2-methyl-benzoimidazol-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]methyl]piperazin-1-yl]tetradecane-1-one (0.012 mg). LC-MS 689.2[M+H] + ,RT 2.44 minutes.

[0171]

[0172] The following paragraphs describe the materials and methods used in conjunction with the data and embodiments described herein. It should be understood that those skilled in the art may use alternative materials and methods generally accepted in the art without departing from the present methods.

[0173] In terms of cell culture and reagents, human breast cancer cell line (MCF-7) was from the American Type Culture Collection (ATCC). hTERT-BJ1 cells were from Clontech, Inc. MCF-7 and hTERT-BJ1 cells were grown in DMEM supplemented with 10% fetal bovine serum, GlutaMAX, and 1% penicillin-streptomycin and incubated at 37°C in a humidified 5% CO2 incubator. The culture medium was changed 2-3 times a week.

[0174] Mammosphere formation assay: Single cell suspensions were prepared using enzymatic disaggregation (1× trypsin-EDTA, Sigma Aldrich, cat. #T3924) and manual disaggregation (25-gauge needle). 5000 cells were plated in mammosphere medium (DMEM-F12 / B27 / 20 ng / ml EGF / PenStrep) in six-well plates coated with 2-hydroxyethyl methacrylate (poly-HEMA, Sigma, cat. #P3932) under non-adherent conditions. Cells were grown for 5 days and maintained in a humidified incubator with 5% (v / v) carbon dioxide / air at 37°C under atmospheric pressure. After 5 days, 3D spheres with a diameter greater than 50 μm were counted using a microscope equipped with a graticule eye-piece, and the percentage of cells that formed spheres was calculated and normalized to 11 (1=100% MFE, i.e., mammosphere formation efficiency). Mammosphere experiments were performed in triplicate and repeated three times independently.

[0175] According to the present method, the selectivity of the compound is characterized using a Hoechst-based viability test to preferentially target cancer cells. In brief, the MCF7 cell monolayer is treated with a compound at a concentration of 1 μM to 100 μM for one day. Hoechst 33342 (a nuclear dye for DNA staining in living cells) is used to assess cell viability. The viability of normal human fibroblasts (hTERT-BJ1) treated with the compounds described herein is also assessed in parallel. Quantification is performed using a plate reader.

[0176] The terms used in describing the embodiments of the present method are only for the purpose of describing specific embodiments and are not intended to be limiting. As used in the specification and the appended claims, the singular forms "a, an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. The present method includes many alternatives, modifications, and equivalents that will become apparent from the following detailed description.

[0177] It should be understood that although the terms "first", "second", "third", "a)", "b)" and "c)" etc. may be used herein to describe the various elements of the present method, the claims should not be limited by these terms. These terms are only used to distinguish one element of the present method from another element. Therefore, the first element discussed below can be referred to as an element aspect, and similarly, the third element can also be referred to as an element aspect without departing from the teachings of the present method. Therefore, the terms "first", "second", "third", "a)", "b)" and "c)" etc. are not intended to convey the necessary order or other hierarchical structure of the related elements, but are only used for identification purposes. The order (or steps) of the operations are not limited to the order presented in the claims.

[0178] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as those commonly understood by those of ordinary skill in the art. It should also be understood that terms such as those defined in common dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the present application and related fields, and unless clearly defined as such in this article, should not be interpreted with an idealized or overly formal meaning. The entire contents of all publications, patent applications, patents and other references mentioned herein are incorporated by reference. In the event of a conflict in terms, this specification shall prevail.

[0179] Furthermore, as used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as no combination ("or") when interpreted in the alternative.

[0180] Unless the context indicates otherwise, it is particularly expected that the various features of the present method described herein can be used in any combination. In addition, the present method also contemplates that, in some embodiments, any feature or feature combination described with respect to the exemplary embodiment can be excluded or omitted.

[0181] As used herein, the transitional phrase "consisting essentially of..." (and grammatical variations) should be interpreted as including the recited materials or steps" as well as those materials or steps that do not materially affect the basic and novel characteristics of the claim. Therefore, as used herein, the term "consisting essentially of..." should not be interpreted as being equivalent to "comprising".

[0182] Having thus described certain embodiments of the present method, it should be understood that the scope of the appended claims is not to be limited to the specific details set forth in the foregoing description, since many obvious variations thereof are apparent without departing from the spirit or scope of what is hereinafter claimed.

Claims

1. A compound comprising the general formula in: R 4 is C3-C8-cycloalkyl; Z is -C(O)NR a R b , where R a and R b are each independently C1-C3-alkyl; and n is an integer from 9 to 20.

2. The compound of claim 1, wherein n is an integer from 12 to 20.

3. The compound of claim 2, wherein R 4 is cyclopentyl, and Z is dimethylformamide.

4. The compound of claim 1, wherein the compound comprises the general formula Wherein n represents an integer from 9 to 20.

5. The compound of claim 4, wherein n is an integer from 12 to 20.

6. The compound of claim 4, wherein n is 12.

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. The pharmaceutical composition of claim 7, wherein the composition is a tablet comprising a core having 35% to 55% by weight of the compound of any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition of claim 8, wherein the pharmaceutically acceptable carrier comprises microcrystalline cellulose, crospovidone type A, low-substituted hydroxypropyl cellulose, magnesium stearate and colloidal anhydrous silicon dioxide.

10. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 in the preparation of a medicament for treating cancer in a patient in need thereof.

11. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 in the preparation of a medicament for treating or preventing metastatic disease in a patient in need thereof.

12. Use of the compound of any one of claims 1-6 or the pharmaceutical composition of any one of claims 7-9 in the preparation of a medicament for treating or preventing tumor recurrence in a patient in need thereof.

13. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 for the preparation of a medicament for reducing resistance to cancer therapy in a patient in need thereof.

14. Use of the compound of any one of claims 1-6 or the pharmaceutical composition of any one of claims 7-9 in the preparation of a medicament for treating or preventing at least one of radiotherapy resistance, chemotherapy resistance and hormone therapy resistance in a patient in need thereof.

15. Use of the compound of any one of claims 1-6 or the pharmaceutical composition of any one of claims 7-9 in the preparation of a medicament for preventing or reducing the proliferation of at least one of cancer cells, cancer stem cells and circulating tumor cells in a patient in need thereof.

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