BAK activators, pharmaceutical compositions and uses in the treatment of cancer - Patents.com

Abstract

The present disclosure relates to an activator of Bak, pharmaceutical composition and use in the treatment of cancer.In certain embodiments, the present disclosure relates to a method for treating cancer, comprising administering an effective amount of the Bak activator disclosed herein to a human subject in need of treatment.In certain embodiments, the present disclosure relates to a pharmaceutical composition comprising the Bak activator 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, ester or salt, and a pharmaceutical acceptable excipient.

Description

[Background technology]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 63 / 224,112, filed July 21, 2021, the entirety of which is incorporated by reference herein for all purposes.

[0002] (STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT) This invention was made with United States Government support under Grant CA200905 awarded by the National Institutes of Health. The United States Government has certain rights in this invention.

[0003] (background) Lung cancer is often classified as non-small cell lung cancer or small cell lung cancer. Non-small cell lung cancer accounts for the majority of lung cancers. Standard treatments for advanced small cell and non-small cell lung cancer include radiation and chemotherapy. Lung cancer is a global health problem. For example, in the United States, more patients die from lung cancer alone than from prostate, breast, and colon cancer combined. Thus, there is a need to identify improved treatments.

[0004] Iyer et al. report that robust autoactivation of apoptosis by BAK, but not BAX, highlights BAK as an important therapeutic target (Cell Death and Disease, 2020, 11:268).

[0005] Kalirajan et al. reported oxazine substituted 9-anilinoacridine derivatives and the evaluation of their antioxidant and anticancer activities (European Journal of Medicinal Chemistry, 2012, 56 217-224).

[0006] Gellerman et al. (WO 2011 / 0519550) report 9-aminoacridine derivatives as potential candidates for the treatment of cancer.

[0007] Park et al. report the discovery of a small molecule Bak activator for the treatment of lung cancer (Theranostics, 2021, 11(17): 8500-8516).

[0008] The references cited herein are not admitted to prior art. Summary of the Invention

[0009] The present disclosure relates to activators of Bak, pharmaceutical compositions and uses in the treatment of cancer. In certain embodiments, the present disclosure relates to a method of treating cancer, comprising administering an effective amount of a Bak activator that is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivatives, prodrugs, esters or salts. In certain embodiments, the present disclosure relates to a method of treating cancer, comprising administering an effective amount of a Bak activator disclosed herein, optionally in combination with other chemotherapeutic agents or therapeutic methods, to a subject in need of treatment.

[0010] In certain embodiments, the subject is a human patient.

[0011] In certain embodiments, the cancer is metastatic cancer, solid cancer or blood cancer.In certain embodiments, the subject is diagnosed with lung cancer, small cell lung cancer or non-small cell lung cancer (NSCLC).In certain embodiments, the subject is diagnosed with cancer selected from breast cancer, colon cancer, lymphoma, multiple myeloma, pancreatic cancer (PANC-1) and osteosarcoma.

[0012] In certain embodiments, the Bak activator disclosed herein is administered in combination with additional chemotherapy.In certain embodiments, the chemotherapy is Bcl-2 inhibitor, such as venetoclax, navitoclax, obatoclax or sabutoclax.In certain embodiments, the chemotherapy is cisplatin, carboplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, gemcitabine, vinorelbine, etoposide, pemetrexed, or combinations thereof.

[0013] In certain embodiments, the chemotherapy agent is a combination of cisplatin or carboplatin, etoposide, paclitaxel or gemcitabine, and vinorelbine.

[0014] In certain embodiments, the disclosure relates to methods of diagnosing and treating a subject with cancer, comprising measuring a Bak level from a sample from the subject; comparing the measured Bak level to a reference or normal value; and administering to the subject an effective amount of a Bak activator, another chemotherapy treatment, a combination chemotherapy treatment, or an aggressive chemotherapy treatment if the measured level is higher than the reference or normal value.

[0015] In certain embodiments, the subject has been diagnosed with a cancer that causes a KRAS mutation, for example, KRAS (G12C, G12D and G12R).

[0016] In certain embodiments, the present disclosure relates to the manufacture of a medicament comprising a Bak activator disclosed herein, such as 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol, a derivative, ester, prodrug or salt thereof, for use in the treatment of cancer.

[0017] In certain embodiments, the present disclosure relates to a pharmaceutical composition comprising the Bak activator disclosed herein or its pharma- ceutically acceptable salt and pharma- ceutically acceptable excipient.In certain embodiments, the pharmaceutical composition is in the form of pill, capsule or tablet.In certain embodiments, the pharmaceutical composition is in the form of isotonic or non-isotonic pH buffered aqueous solution. [Brief description of the drawings]

[0018] [Figure 1A] FIG. 1A shows the chemical structure of Bak activator-073 (BKA-073), which has the chemical name 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol. [Figure 1B] Figure 1B shows data indicating that BKA-073 is a compound that targets the BH3 domain of Bak and induces mitochondrial priming and apoptosis in lung cancer cells. The expression levels of Bak in NSCLC and SCLC cell lines were analyzed by Western blot. A panel of NSCLC and SCLC cell lines was treated with BKA-073 (1 μM) for 16 or 72 hours, and then analyzed for dynamic BH3 profiling or apoptotic cell death. [Figure 1C] Figure 1C shows data showing that BKA-073 induces mitochondrial priming and apoptosis. The expression levels of Bak in various types of cancer cell lines were analyzed by Western blot. Cancer cell lines were treated with BKA-073 (1 μM) for 16 or 72 hours, and then analyzed for dynamic BH3 profiling or apoptotic cell death. [Figure 2A] Figures 2A-2B show data demonstrating that BKA-073 specifically binds to Bak and induces Bak oligomerization. Figure 2A shows data from a fluorescence polarization assay performed to measure inhibition constants (Ki) using purified Bak protein or other Bcl2 family members, BKA-073, and fluorescently labeled BakBH3 peptide. [Figure 2B]Figure 2B shows data on the binding affinity of WT Bak or ΔBH3 Bak deletion mutant proteins with BKA-073, as tested by isothermal titration calorimetry assay. Binding constant (KD) values ​​were determined by fitting the titration curves to a one-site binding mode. [Diagram 3] Figure 3 shows data demonstrating that BKA-073 potently inhibits lung cancer growth in a dose-dependent manner in vivo. Nu / Nu mice bearing A549 lung cancer xenografts were treated intraperitoneally with increasing doses of BKA-073 (5-15 mg / kg / day) for 28 days. Tumor volumes were measured every 2 days. After treatment, mice were sacrificed and tumors were excised and analyzed. [Figure 4] Figure 4 shows data showing that BKA-073 suppresses SCLC in xenograft and PDX models. Nu / Nu mice bearing SCLC cell line DMS114 or xenografts from SCLC patients (TKO-2 or TKO-5) were intraperitoneally treated with BKA-073 (15 mg / kg / day) for 14 or 28 days. Tumor volumes were measured every 2 days. After treatment, mice were sacrificed and tumors were excised and analyzed. [Diagram 5] Figure 5 shows data demonstrating that BKA-073 extends survival in genetically engineered mouse models (GEMMs). KRAS G12D LKB1fl / fl (KL) mice were administered adenoviral Cre recombinase for 6 weeks, after which the KL mice were treated intraperitoneally with KRA-073 (15 mg / kg / day) for 48 days (n=6 mice / group). Mice survival is shown up to 48 days before euthanasia in the control and BKA-073-treated groups. [Figure 6]Figure 6 shows data demonstrating that BKA-073 synergizes with the Bcl-2 inhibitor ABT-199 (venetoclax) against SCLC and NSCLC in vitro and in vivo. Nu / Nu mice bearing SCLC DMS53 xenografts or NSCLC H460 xenografts were treated with BKA-073 (10 mg / kg / day) intraperitoneally, ABT-199 (60 mg / kg / day) orally, or the combination for 28 days. Tumor volumes were measured every 2 days. After treatment, mice were sacrificed and tumors were excised and analyzed. [Figure 7] Figure 7 shows data indicating that high levels of Bak expression are associated with poor prognosis in NSCLC patients. Kaplan-Meier survival curves for NSCLC patients, n=208. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Before describing the present invention in more detail, it is to be understood that this disclosure is not limited to described embodiments, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.

[0021] All publications and patents cited herein are incorporated by reference to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference, and are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by prior disclosure. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0022] "Embodiments" refer to examples and are not necessarily limited to such examples. The embodiments of the present disclosure employ, unless otherwise indicated, techniques of medicine, organic chemistry, biochemistry, molecular biology, pharmacology, and the like, which are within the skill of the art. Such techniques are fully explained in the literature.

[0023] As will be apparent to one of ordinary skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the disclosure. Any recited method may be carried out in the order of events recited or in any other order which is logically possible.

[0024] To the extent that any chemical formula given herein contains one or more chiral centers, the formula is intended to encompass all stable stereoisomers, enantiomers, and diastereomers, and the formula is also understood to encompass all tautomers.

[0025] It must be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. In this specification and the claims that follow, reference will be made to a number of terms that will be defined to have the following meanings unless a contrary intention is apparent.

[0026] "Bak", also referred to as "Bcl-2 homologous antagonist / killer", is a pore-forming proapoptotic protein that contains a BH3 domain and is therefore classified as a BCL-2 family protein. BCL2 family members form oligomers or heterodimers and function as regulators of various cellular activities. Bak has been reported to activate apoptosis within mitochondria. Human [Homo sapiens] Bcl-2 homologous antagonist / killer is designated as NCBI Reference Sequence: NP_001179.1.

[0027] As used herein, a "subject" refers to any animal, preferably a human patient, livestock or household pet.

[0028] As used herein, the terms "treat" and "treating" are not limited to cases where a subject (e.g., a human patient) is cured and the disease is eradicated. Rather, embodiments of the present disclosure also contemplate treatment that merely alleviates symptoms and / or slows the progression of the disease.

[0029] As used herein, the term "in combination with," when used to describe administration with a further treatment, means that the agent can be administered prior to, simultaneously with, or after the further treatment, or a combination thereof.

[0030] " Salt " as used herein refers to the derivative of disclosed compounds, which parent compound is modified to form its acid salt or base salt.Examples of salt include, but are not limited to, mineral or organic acid salts of basic residues such as amine, alkylamine or dialkylamine; alkali or organic salts of acidic residues such as carboxylic acid.In certain embodiments, salt is a conventional non-toxic pharmaceutically acceptable salt, including the quaternary ammonium salt of parent compound formed, and non-toxic inorganic or organic acid.

[0031] The term "derivative" as used herein refers to a structurally similar compound that retains the full functional properties of the identified analog. A derivative may be structurally similar because it lacks one or more atoms, is substituted, is a salt, has a different hydration / oxidation state, or has one or more atoms in the molecule replaced, such as, but not limited to, replacing an oxygen atom with a sulfur atom or replacing an amino group with a hydroxyl group. A derivative may be a prodrug. Derivatives may be produced by various synthetic methods or suitable modification methods described in synthesis or organic chemistry textbooks, such as those shown in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze, which are incorporated herein by reference.

[0032] The term "substituted" refers to a molecule in which at least one hydrogen atom has been replaced with a substituent. When substituted, one or more of the group is a "substituent." A molecule may be multiply substituted. In the case of an oxo substituent (=O), two hydrogen atoms are replaced. Examples of substituents in this context are halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NR a R b , -NR a C(=O)R b , -NR a C(=O)NR a NR b , -NR a C(=O)OR b , -NR a SO 2 R b , -C(=O)R a , -C(=O)OR a , -C(=O)NR a R b , -OC(=O)NR a R b , -OR a , -SR a , -SOR a , -S(=O) 2 R a , -OS(=O) 2 R a and -S(=O) 2 OR a In this regard, R a and R b can be the same or different and can independently be hydrogen, halogen hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, or heteroarylalkyl.

[0033] The term "prodrug" as used herein refers to a compound that is metabolized (i.e., transformed in the body) into a pharmacologically active drug after administration. Examples include alkoxy esters of hydroxyl or carboxyl groups, such as acetates, benzoates, alkyl ethers, amino acid esters, glycolates, malates, acyloxyalkyl esters, alkoxycarbonyloxyalkyl esters, S-acylthioalkyl esters, hydroxylamine amides, phosphonylmethoxy ethers, phosphates, phosphoramidates, and combinations thereof.

[0034] Prodrugs may also have improved solubility in pharmaceutical compositions compared to the parent drug. Prodrugs may be converted to the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Typical prodrugs are pharma-ceutically acceptable esters. Prodrugs include compounds in which hydroxy, amino or mercapto groups are bonded to any group that is cleaved to form a free hydroxy, free amino or free mercapto group, respectively, when the prodrug of the active compound is administered to a subject.

[0035] When a disclosed compound or a pharma- ceutically acceptable form of said compound contains an alcohol functional group, the prodrug may be a prodrug that converts a hydrogen atom of the alcohol group to a hydrogen atom of (C 1 -C 6 )(alkanoyloxy)methyl, 1-((C 1 -C 6 )alkanoyloxy)ethyl, 1-methyl-1((C 1 -C 6 )Alkanoyloxy)ethyl(C 1 -C 6 )(alkoxycarbonyloxy)methyl, N-(C 1 -C 6 ) alkoxycarbonylaminomethyl, succinoyl, (C 1 -C 6 ) Alkanoyl, α-amino (C 1 -C 4) alkanoyl, aryl acyl and α-amino acyl, or α-amino acyl-α-amino acyl groups, where each α-amino acyl group independently corresponds to the -P(O)(OH) of a natural L-amino acid. 2 , -P(O)(O(C 1 -C 6 )Alkyl) 2 and glycosyl (the radical resulting from removal of the hydroxyl group of the hemiacetal form of a carbohydrate).

[0036] When a disclosed compound, or a pharma- ceutically acceptable form of the compound, contains an amine functional group, a prodrug can be formed by replacing a hydrogen atom of the amine group with a group such as R-carbonyl, RO-carbonyl, NRR′-carbonyl, and the like, where R and R′ are each independently: 1 -C 10 ) alkyl, (C 3 -C 7 ) Cycloalkyl, benzyl, natural α-aminoacyl, -C(OH)C(O)OY 1 (In the formula, Y 1 is H, (C 1 -C 6 ) alkyl or benzyl), -C(OY 2 )Y 3 (In the formula, Y 2 is (C 1 -C 4 ) alkyl, and Y 3 is (C 1 -C 6 ) alkyl, carboxy (C 1 -C 6 ) Alkyl, Amino (C 1 -C 4 ) alkyl or mono-N or di-N,N-(C 1 -C 6 ) alkylaminoalkyl), -C(Y 4 )Y 5 (In the formula, Y 4 is H or methyl, and Y 5 is mono-N- or di-N,N-(C 1 -C 6) alkylamino), morpholino, piperidin-1-yl or pyrrolidin-1-yl.

[0037] As used herein, "alkyl" refers to an acyclic straight or branched chain unsaturated or saturated hydrocarbon, for example, containing 1 to 25 carbon atoms. For example, "C 8 -C 18 " refers to an alkyl group containing 8 to 18 carbon atoms. 6 -C 22 " refers to an alkyl containing 6 to 22 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl, n-nonyl, and the like; while saturated branched chain alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (referred to as "alkenyl" or "alkynyl", respectively). Representative straight chain and branched chain alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; while representative straight chain and branched chain alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, and the like.

[0038] Non-aromatic monocyclic or polycyclic alkyls are referred to herein as "carbocycle" or "carbocyclyl" groups. Representative saturated carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like; while unsaturated carbocycles include cyclopentenyl and cyclohexenyl, and the like.

[0039] A "heterocarbocycle" or "heterocarbocyclyl" group is a carbocycle which may be saturated or unsaturated (not aromatic), monocyclic or polycyclic, containing from one to four heteroatoms selected from nitrogen, oxygen and sulfur, where the nitrogen and sulfur heteroatoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. Heterocarbocycles include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

[0040] The term "aryl" refers to an aromatic homocyclic (i.e., hydrocarbon) monocyclic, bicyclic, or tricyclic ring-containing group, preferably having 6 to 12 members, such as phenyl, naphthyl, and biphenyl. Phenyl is a preferred aryl group.

[0041] As used herein, "heteroaryl" or "heteroaromatic" refers to an aromatic heterocarbocycle having from 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur and containing at least one carbon atom, including both monocyclic and polycyclic ring systems. Polycyclic ring systems may, but need not, contain one or more non-aromatic rings, so long as one of the rings is aromatic. Representative heteroaryls are furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl. Use of the term "heteroaryl" is intended to include N-alkylated derivatives, such as a 1-methylimidazol-5-yl substituent.

[0042] As used herein, "heterocycle" or "heterocyclyl" refers to monocyclic and polycyclic ring systems having from 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur and containing at least one carbon atom. The monocyclic and polycyclic ring systems can be aromatic, non-aromatic, or mixtures of aromatic and non-aromatic rings. Heterocycles include heterocarbocycles, heteroaryls, and the like.

[0043] "Alkoxy" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through an oxygen bridge. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, n-pentoxy and s-pentoxy. Preferred alkoxy groups are methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy.

[0044] "Alkoxyalkyl" refers to an alkyl group as defined above having the indicated number of carbon atoms attached through an alkyl bridge (i.e., -CH 2 -O-CH 2 CH 3 )

[0045] "Alkylamino" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through an amino bridge. An example of an alkylamino is methylamino (i.e., -NH-CH 3 ).

[0046] "Alkylthio" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through a sulfur bridge. An example of an alkylthio group is methylthio (i.e., -S-CH 3 ).

[0047] "Alkanoyl" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through a carbonyl bridge (ie, --(C.dbd.O)alkyl).

[0048] The terms "cycloalkyl" and "cycloalkenyl" refer to mono-, bi- or tricyclic homocyclic radicals of 3 to 15 carbon atoms that are fully saturated and partially unsaturated, respectively.

[0049] "Alkylsulfonyl" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through a sulfonyl bridge (i.e., -S(=O) 2 "Arylsulfonyl" refers to an aryl (i.e., -S(=O)) bonded through a sulfonyl bridge. 2 (refers to aryl).

[0050] "Alkylsulfamoyl" refers to an alkyl group as defined above with the indicated number of carbon atoms attached through a sulfamoyl bridge (i.e., -NHS(=O) 2 "Arylsulfamoyl" refers to an alkyl group attached via a sulfamoyl bridge (i.e., -NHS(=O) 2 Aryl)

[0051] "Alkylsulfinyl" refers to an alkyl as defined above with the indicated number of carbon atoms attached through a sulfinyl bridge (ie, --S(.dbd.O)alkyl).

[0052] The terms "halogen" and "halo" refer to fluorine, chlorine, bromine and iodine.

[0053] In certain embodiments, the disclosure contemplates the compounds or compositions disclosed herein in the manufacture of a medicament for use in the treatment of cancer. "Cancer" refers to any of a variety of cellular diseases involving malignant neoplasms characterized by cell proliferation. It is not intended that diseased cells must actually invade surrounding tissues and metastasize to new body sites. Cancer may involve any tissue of the body and take various forms in each area of ​​the body. Within the context of certain embodiments, whether "cancer has been alleviated" may be identified by various diagnostic methods known to those skilled in the art, including, but not limited to, observing a decrease in the size or number of tumor masses, or observing an increase in apoptosis of cancer cells, for example, whether apoptosis of cancer cells is increased by more than 5% for the sample compound compared to a control without the compound. It may also be identified by changes in relevant biomarkers or gene expression profiles, such as PSA for prostate cancer and HER2 for breast cancer.

[0054] The cancer to be treated in the context of this disclosure can be any kind of cancer or tumor, such as lung cancer, non-small cell lung cancer and subtypes of NSCLC, such as adenocarcinoma, squamous cell carcinoma and large cell carcinoma, and small cell lung cancer. Malignant tumors present in the colon, abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal glands, parathyroid gland, pituitary gland, testis, ovary, thymus, thyroid gland), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvis, skin, soft tissue, spleen, chest and urogenital tract, more specifically, adrenal cortical carcinoma, AIDS-related lymphoma, AIDS-related malignant tumor, anal cancer, astrocytoma, biliary tract cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, renal pelvis and ureter cancer, primary central nervous system cerebellar astrocytoma, brain astrocytoma, cervical cancer, chronic lymphocytic white tumor. Hematologic malignancies, chronic myeloid leukemia, colon cancer, cutaneous T-cell lymphoma, endocrine islet cell carcinoma, endometrial cancer, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's sarcoma and related tumors, exocrine pancreatic cancer, extracranial germ cell tumors, extragonadal germ cell tumors, extrahepatic biliary tract cancer, eye cancer, Gaucher's disease, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal tumors, germ cell tumors, gestational trophoblastic tumors, head and neck cancer, hepatocellular carcinoma, hypergammaglobulinemia, hypopharyngeal cancer, Hodgkin's disease, intestinal cancer, intraocular melanoma, islet cell carcinoma, islet cell carcinoma, Kaposi's sarcoma, laryngeal cancer, lip cancer, macroglobulinemia , malignant mesothelioma, malignant thymoma, medulloblastoma, melanoma, mesothelioma, occult primary metastatic squamous cell neck cancer, primary metastatic squamous cell neck cancer, metastatic squamous cell neck cancer, multiple myeloma, multiple myeloma / plasma cell neoplasm, myelodysplastic syndrome, myeloid leukemia, myeloproliferative disorders, paranasal sinus and nasal cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-melanoma skin cancer, non-small cell lung cancer, metastatic squamous cell neck cancer of unknown primary, buccopharyngeal carcinoma, malignant fibrous histiocytoma, malignant fibrous osteosarcoma / histiocytoma of bone, epithelial ovarian cancer, ovarian germ cell tumor, ovarian Focal low-grade malignant tumors, pancreatic cancer, paraproteinemia, purpura, parathyroid cancer, penile cancer, pituitary tumors, plasma cell neoplasms / multiple myeloma, primary central nervous system lymphoma, primary liver cancer, prostate cancer, rectal cancer, renal cell carcinoma, carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoidosis, sarcoma, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell cervical cancer, gastric cancer, pineal and supratentorial primitive neuroectodermal tumors, T-cell lymphoma, testicular cancer, thymoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, transitional carcinoma of the renal pelvis and ureter,Trophoblastic tumors, renal pelvic and ureteral cell carcinomas, urethral carcinoma, uterine carcinoma, uterine sarcoma, vaginal carcinoma, optic nerve pathway and hypothalamic gliomas, vulvar carcinoma, Waldenstrom's macroglobulinemia, Wilms' tumor and other hyperproliferative disorders, as well as neoplasms present in the aforementioned organ systems, are contemplated.

[0055] In certain embodiments, the compound may be administered in combination with an additional anticancer drug. "Chemotherapy agent", "chemotherapeutic", "anticancer drug" and the like refer to molecules recognized to be useful in the treatment of cancer. Examples contemplated include the following molecules or derivatives, such as abemaciclib, abiraterone acetate, methotrexate, paclitaxel, adriamycin, acalabrutinib, brentuximab vedotin, adotrastuzumab emtansine, aflibercept, afatinib, netupitant, palonosetron, imiquimod, aldesleukin, alectinib, pemetrexed disodium, copanlisib, melphalan, brigatinib, chlorambucil, amifostine, aminolevulinic acid, ana. Strozole, apalutamide, aprepitant, pamidronate disodium, exemestane, nelarabine, arsenic trioxide, ofatumumab, atezolizumab, bevacizumab, avelumab, axicabtagene ciloreucel, axitinib, azacitidine, carmustine, belinostat, bendamustine, inotuzumab ozogamicin, bevacizumab, bexarotene, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib , busulfan, irinotecan, capecitabine, fluorouracil, carboplatin, carfilzomib, ceritinib, daunorubicin, cetuximab, cisplatin, cladribine, cyclophosphamide, clofarabine, cobimetinib, cabozantinib-S-malate, dactinomycin, crizotinib, ifosfamide, ramucirumab, cytarabine, dabrafenib, dacarbazine, decitabine, daratumumab, dasatinib, defibrotide, degarelix, denileukin diftitoc s, denosumab, dexamethasone, dexrazoxane, dinutuximab, docetaxel, doxorubicin, durvalumab, rasburicase, epirubicin, elotuzumab, oxaliplatin, eltrombopag olamine, enasidenib, enzalutamide, eribulin, vismodegib, erlotinib, etoposide, everolimus, raloxifene, toremifene, panobinostat, fulvestrant, letrozole, filgrastim, fludarabine, flutamide, pralatrexate,Obinutuzumab, gefitinib, gemcitabine, gemtuzumab ozogamicin, glucarpidase, goserelin, propranolol, trastuzumab, topotecan, palbociclib, ibritumomab tiucetan, ibrutinib, ponatinib, idarubicin, idelalisib, imatinib, talimogenera herparepvec, ipilimumab, romidepsin, ixabepilone, ixazomib, ruxolitinib, cabazitaxel, palifermin, pembrolizumab, ribociclib, tisagenlucrucel, lanreotide, lapatinib, olaratumab, lenalidomide, Lenvatinib, leucovorin, leuprolide, lomustine, trifluridine, olaparib, vincristine, procarbazine, mechlorethamine, megestrol, trametinib, temozolomide, methylnaltrexone bromide, midostaurin, mitomycin C, mitoxantrone, plerixafor, vinorelbine, necitumumab, neratinib, sorafenib, nilutamide, nilotinib, niraparib, nivolumab, tamoxifen, romiplostim, sonidegib, omacetaxine, pegaspargase, ondansetron, osimertinib, panitumumab , pazopanib, interferon alpha-2b, pertuzumab, pomalidomide, mercaptopurine, regorafenib, rituximab, rolapitant, rucaparib, siltuximab, sunitinib, thioguanine, temsirolimus, thalidomide, thiotepa, trabectedin, valrubicin, vandetanib, vinblastine, vemurafenib, vorinostat, zoledronic acid, or combinations thereof, such as cyclophosphamide, methotrexate, 5-fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine rituximab, cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); rituximab, cyclophosphamide, doxorubicin, vincristine, prednisolone (RCHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5-fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine,doxorubicin, cisplatin (MVAC). In certain embodiments, the chemotherapeutic agent is an antibody, an anti-PD-1, an anti-PD-L1, an anti-CTLA4 antibody, or a combination thereof, such as an anti-CTLA4 (e.g., ipilimumab, tremelimumab), an anti-PC-L1 (e.g., atezolizumab, avelumab, durvalumab) or an anti-PD1 antibody (e.g., nivolumab, pembrolizumab, cemiplimab, dostarlimab, spartalizumab, camrelizumab, tislelizumab, toripalimab, sintilimab).

[0056] Bak Activator Although it is not intended that particular embodiments of the present disclosure be limited by a particular mechanism, certain compounds disclosed herein are believed to activate Bak; therefore, the compounds are useful as therapeutic agents for treating cancer.

[0057] In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), a derivative, prodrug, ester or salt thereof. In certain embodiments, the derivative has Formula I or II: [ka] [In the formula, Q is O or S; U is N or CH; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are each individually and independently hydrogen, alkyl, halogen, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkanoyl, alkylthio, alkylamino, aminoalkyl, (alkyl) 2amino, phosphate, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, where R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and R 10 is one or more identical or different R 11 may be substituted with; R 11 is alkyl, halogen, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkanoyl, alkylthio, alkylamino, phosphate, aminoalkyl, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, where R 11 is one or more identical or different R 12 may be substituted with; R 12 is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, 2-methoxyethoxy, 2-hydroxyethoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl or heterocyclyl] or a derivative, prodrug, ester or salt thereof.

[0058] In certain embodiments, R 1 is hydrogen. In certain embodiments, R 2 is alkyl. In certain embodiments, R 3 is hydrogen. In certain embodiments, R 4 is hydrogen. In certain embodiments, R 5 is alkyl or methyl. In certain embodiments, R 6 , R 7 , R 8 , R 9 and R 10 is hydrogen. In certain embodiments, Q is O. In certain embodiments, U is NH.

[0059] Pharmaceutical Compositions In certain embodiments, the present disclosure relates to a pharmaceutical composition comprising a Bak activator disclosed herein and a pharma- ceutically acceptable excipient. In certain embodiments, the pharma-ceutically acceptable excipient is selected from a diluent, a disintegrant, a solubilizer, or a lubricant.

[0060] In certain embodiments, the pharma- ceutically acceptable excipient is selected from monosaccharides, disaccharides, sucrose, lactose, glucose, mannitol, sorbitol, polysaccharides, starch, cellulose, microcrystalline cellulose, cellulose ethers, hydroxypropylcellulose (HPC), xylitol, maltitol, gelatin, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), hydroxypropylmethylcellulose (HPMC), cross-linked sodium carboxymethylcellulose, dicalcium phosphate, calcium carbonate, stearic acid, magnesium stearate, talc, magnesium carbonate, silica, vitamin A, vitamin E, vitamin C, retinyl palmitate, cerium, cysteine, methionine, citric acid and sodium citrate, methylparaben, propylparaben, and combinations thereof.

[0061] In certain embodiments, pharma- ceutically acceptable additive is diluent.Examples include crystalline cellulose, and other diluents can be, for example, calcium carbonate, calcium phosphate, calcium sulfate, sodium acetate, erythritol, ethylcellulose, fructose, inulin, isomalt, lactitol, lactose, magnesium carbonate, magnesium oxide, maltitol, maltodextrin, maltose, mannitol, polydextrose, polyethylene glycol, pullulan, simethicone, sodium bicarbonate, sodium carbonate, sodium chloride, sorbitol, starch, sucrose, trehalose and xylitol.

[0062] In certain embodiments, the pharma- ceutically acceptable additive is disintegrant.The example of disintegrant can be, for example, alginic acid, calcium alginate, carboxymethylcellulose calcium, chitosan, colloidal silicon dioxide, croscarmellose sodium, crospovidone, glycine, guar gum, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, magnesium aluminum silicate, methylcellulose, povidone, sodium alginate, sodium carboxymethylcellulose, sodium starch glycolate and starch.

[0063] In certain embodiments, the pharma- ceutically acceptable additive is solubilizer.The example of solubilizer can be, for example, benzalkonium chloride, benzyl benzoate, sulfobutyl ether beta-cyclodextrin sodium, cetylpyridinium chloride, cyclodextrin, diethylene glycol monoethyl ether, fumaric acid, hydroxypropyl beta-cyclodextrin, hypromellose, lanolin alcohol, lecithin, oleyl alcohol, phospholipid, poloxamer, polyoxyethylene alkyl ether, polyoxyethylene castor oil derivative, polyoxyethylene sorbitan fatty acid ester, polyoxyl hydroxystearic acid polyoxyl, polyoxyl glyceride, povidone, pyrrolidone, sodium lauryl sulfate, sorbitan ester (sorbitan fatty acid ester), tricaprylin, triolein and vitamin E polyethylene glycol succinate.

[0064] In certain embodiments, the pharma- ceutically acceptable additive is lubricant.The example of lubricant can be, for example, calcium stearate, glyceryl behenate, glyceryl dibehenate, glyceryl monostearate, glyceryl palmitostearate, mixture of behenic acid esters of glycerin (for example, mixture of glyceryl dibehenate, tribehenin and glyceryl behenate), leucine, magnesium stearate, myristic acid, palmitic acid, poloxamer, polyethylene glycol, potassium benzoate, sodium benzoate, sodium lauryl sulfate, sodium stearate, sodium stearyl fumarate, stearic acid, talc, tribehenin and zinc stearate.

[0065] In certain embodiments, the pharma- ceutically acceptable excipient is selected from lactose, sucrose, mannitol, triethyl citrate, dextrose, cellulose, methylcellulose, ethylcellulose, hydroxylpropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, croscarmellose sodium, polyvinyl N-pyrrolidone, crospovidone, ethylcellulose, povidone, methyl and ethyl acrylate copolymers, polyethylene glycol, fatty acid esters of sorbitol, lauryl sulfate, gelatin, glycerin, glyceryl monooleate, silicon dioxide, titanium dioxide, talc, corn starch, carnauba wax, stearic acid, sorbic acid, magnesium stearate, calcium stearate, castor oil, mineral oil, calcium phosphate, starch, carboxymethyl ether of starch, iron oxide, triacetin, acacia gum, esters or salts thereof.

[0066] In certain embodiments, the pharmaceutical composition is in the form of a tablet, a pill, a capsule, a gel, a gel capsule or a cream.In certain embodiments, the pharmaceutical composition is in the form of a sterile pH buffered aqueous saline solution or a physiological saline phosphate buffer, optionally containing a monosaccharide or monosaccharide, at pH 6-8.

[0067] In certain embodiments, the pharma- ceutically acceptable form is a pharma-ceutically acceptable salt.As used herein, the term "pharma-ceutically acceptable salt" refers to a salt that is suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic reaction, etc., within the scope of sound medical judgment, and is commensurate with a reasonable benefit / risk ratio.Pharmaceutically acceptable salts are well known in the art.For example, Berge et al. describe pharma-ceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19.Pharmaceutically acceptable salts of the compounds provided herein are derived from inorganic and organic acids and bases.Examples of pharma-ceutically acceptable non-toxic acid addition salts are the salts of amino groups formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, and with acids or organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange.

[0068] Other pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, Examples of suitable salts include lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. In some embodiments, organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0069] Pharmaceutically acceptable salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, or N + (C 1-4 Alkyl) 4Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations, which are formed with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates and arylsulfonates, where appropriate. Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium and magnesium salts.

[0070] In certain embodiments, the Bak activator disclosed herein can be used in "free base form" or as a pharma- ceutically acceptable salt, or any mixture thereof. In one embodiment, the Bak activator is in free base form. "Free base form" is understood to refer to the case where the Bak activator is not in the form of a salt.

[0071] In certain embodiments, the present disclosure relates to a kit or pharmaceutical packaging, comprising the Bak activator or drug combination disclosed herein with instructions for use.In certain embodiments, each drug can be packaged in a container, such as a vial, box, syringe or bottle.In certain embodiments, instructions can be in a pamphlet in the container, or on the outside or inside of the container.

[0072] How to use In certain embodiments, the present disclosure relates to a method for treating cancer, comprising administering an effective amount of a Bak activator or a pharmaceutical composition comprising the same to a subject in need of treatment.In certain embodiments, the subject is a human patient.In certain embodiments, the Bak activator induces or increases cell apoptosis, for example, the formation of Bak oligomers in mitochondria promotes cytochrome c (Cyt c) release to induce apoptosis.In certain embodiments, the present disclosure relates to the manufacture of a medicament comprising the Bak activator disclosed herein for use in treating cancer.

[0073] In certain embodiments, the cancer is metastatic cancer, solid cancer or blood cancer.In certain embodiments, the subject is diagnosed with lung cancer, small cell lung cancer or non-small cell lung cancer (NSCLC).In certain embodiments, the subject is diagnosed with cancer selected from breast cancer, colon cancer, lymphoma, multiple myeloma, pancreatic cancer (PANC-1) and osteosarcoma.

[0074] In certain embodiments, the subject is diagnosed with cancer selected from lung cancer, pancreatic cancer, colorectal cancer, uterine cancer, esophageal cancer, gastric cancer, cervical cancer, breast cancer, prostate cancer or bladder cancer.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0075] In certain embodiments, the Bak activator is administered in combination with an additional chemotherapeutic agent. In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0076] In certain embodiments, the subject is diagnosed with non-small cell lung cancer (NSCLC). In certain embodiments, the malignant cells are found in sputum cytology. In certain embodiments, the tumor can be found by bronchoscopy or imaging.

[0077] In certain embodiments, the therapeutic methods disclosed herein may be administered in addition to surgery to remove a portion of the lung, such as a lobectomy, sleeve resection, segmentectomy, or wedge resection.

[0078] In certain embodiments, the therapeutic approaches disclosed herein may be administered in addition to radiation therapy.

[0079] In certain embodiments, a Bak activator disclosed herein is administered in combination with an additional chemotherapeutic agent.

[0080] In certain embodiments, the chemotherapeutic agent is a Bcl-2 inhibitor, such as venetoclax, navitoclax, obatoclax, or sabutoclax.

[0081] In certain embodiments, the chemotherapeutic agent is cisplatin, carboplatin, paclitaxel, albumin-bound paclitaxel, docetaxel, gemcitabine, vinorelbine, etoposide, pemetrexed, or a combination thereof.

[0082] In certain embodiments, the chemotherapy agent is a combination of cisplatin or carboplatin, gemcitabine, and vinorelbine or paclitaxel.

[0083] In certain embodiments, the disclosure relates to a method of treating leukemia comprising administering to a subject in need of treatment an effective amount of a Bak activator, such as 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), a derivative, prodrug or salt thereof, in combination with venetoclax or other Bcl-2 inhibitor.

[0084] In certain embodiments, the present disclosure relates to a method of treating leukemia, comprising administering to a subject in need of treatment an effective amount of a Bak activator, such as 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt, in combination with rituximab and venetoclax or other Bcl-2 inhibitor. In certain embodiments, the subject has been diagnosed with chronic lymphocytic leukemia (CLL), or relapsed or refractory chronic lymphocytic leukemia (CLL). In certain embodiments, the subject has been diagnosed with acute myeloid leukemia (AML) for treatment with a Bak activator, such as 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), a derivative, prodrug or salt thereof, in combination with a hypomethylating agent, such as decitabine and azacitidine, or cytarabine.

[0085] In certain embodiments, the cancers disclosed herein may be introduced into a subject who has been diagnosed with a genetic mutation, such as a mutation in an Akt, Mcl-1, EGFR, ALK, ROS1, BRAF, RET, MET, NTRK gene, or a combination thereof.

[0086] In certain embodiments, the subject is diagnosed with Akt gene mutation (e.g., L52R, Q79K and D323H).In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with an Akt inhibitor, such as capivasertib and ipatasertib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0087] In certain embodiments, the subject is diagnosed with Mcl-1 gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with the Mcl-1 inhibitor disclosed herein.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0088] In certain embodiments, the subject is diagnosed with ALK gene mutation.In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with an ALK inhibitor.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with crizotinib, alectinib, brigatinib, lorlatinib, foretinib, albotinib, verizatinib, repotrectinib, entrectinib or ensartinib.

[0089] In certain embodiments, the subject is diagnosed with EGFR gene mutation.In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with an EGFR inhibitor.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with afatinib, erlotinib or lapatinib.

[0090] In certain embodiments, the subject is diagnosed with ROS1 gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with crizotinib, entrectinib or ceritinib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0091] In certain embodiments, the subject is diagnosed with BRAF gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with dabrafenib or trametinib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0092] In certain embodiments, the subject is diagnosed with RET gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with selpercatinib or pralsetinib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0093] In certain embodiments, the subject is diagnosed with MET gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with capmatinib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0094] In certain embodiments, the subject is diagnosed with NTRK gene mutation.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with larotrectinib or entrectinib.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0095] In certain embodiments, the subject is diagnosed with a tumor or cancer cells that have higher than normal levels of PD-L1.In certain embodiments, the subject is administered a Bak activator disclosed herein in combination with a PD-L1 antibody, pembrolizumab, atezolizumab, nivolumab or ipilimumab.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0096] In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with bevacizumab to treat cancer.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt.

[0097] In certain embodiments, the subject is diagnosed with squamous cell NSCLC.In certain embodiments, the subject is administered the Bak activator disclosed herein in combination with necitumumab.In certain embodiments, the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, prodrug or salt. EXAMPLES

[0098] Bak activators for cancer treatment Bak is a proapoptotic protein required for the induction of programmed cell death and apoptosis in cancer cells. Experiments reported herein show that elevated Bak expression correlates with poor prognosis of lung cancer, indicating that Bak is a promising prognostic indicator and potential therapeutic target in lung cancer patients. Here, BKA-073 was identified as a Bak activator that targets the BH3 domain of Bak, activates the proapoptotic function of Bak, and exhibits potent antitumor activity against lung cancer and other cancers. Experiments show that BKA-073 directly binds to Bak protein and induces Bak oligomerization in mitochondria, causing activation of proapoptotic function. Experiments show that BKA-073-induced Bak oligomerization promotes mitochondrial priming and Cyt c release, which are early changes in net proapoptotic signaling at mitochondria. BKA-073-induced mitochondrial priming and Cyt c release trigger apoptotic cell death of lung cancer cells. Knockout of Bak, but not Bax, results in BKA-073 resistance in lung cancer cells and lung cancer xenografts, indicating that the antitumor activity of BKA-073 occurs in a Bak-dependent manner. Furthermore, exogenous expression of wild-type Bak, but not deleted BH3 mutants, in A549Bak double-negative cells can restore sensitivity to BKA-073, indicating that the apoptotic effect of BKA-073 involves binding to the BH3 domain of the Bak protein. Although it is not intended that the embodiments of the present disclosure be limited by a particular mechanism, the experimental results suggest a mechanistic model for the use of small molecule Bak activators in cancer therapy.

[0099] Several Bax / Bak-independent mechanisms of apoptotic cell death have been reported. The experiments reported here show that a small percentage (approximately 20%) of apoptotic cell death was observed in Bak- / - and DKOA549 cells. In addition to the predominant Bak-dependent apoptotic mechanism, BKA-073 appears to induce a small percentage (approximately 20%) of apoptotic cell death via a Bax / Bak-independent mechanism.

[0100] BKA-073 demonstrated potent antitumor activity against lung cancer through induction of Bak activation (oligomerization) and apoptotic cell death in xenografts derived from lung cancer cell lines or patient-derived SCLC tumors. A dose range of 5-15 mg / kg / day was effective without weight loss or significant organ toxicity. BKA-073 inhibited the growth of patient-derived xenografts (PDXs) from two patients with refractory SCLC, suggesting that BKA-073 may have clinical utility in human patients.

[0101] Screening for small molecules targeting the BH3-binding pocket of Bak The BH3 death domain is required for the proapoptotic function of Bak. The BH3 domain binding pocket (aa75-88) of Bak (PDB ID: 2YV6) was selected as a docketing site for screening small molecules using the UCSF DOCK 6.1 program suite and the NCI chemical library (300,000 small molecules) database. The small molecules were ranked according to their energy scores. The top 500 compounds determined to have the highest affinity for the BH3 domain were obtained from NCI and tested for cytotoxicity in human lung cancer cells (H1299, H460 and A549 cells) by sulforhodamine B (SRB) assay for further screening. Among these small molecules, compound NSC14073 had the most potent activity against human lung cancer cells. This Bak activator compound was compared with BKA-073 (C 19 H 24 ClN 3 O 2, MW: 361.87) (Figure 1A).

[0102] To test the effect of BKA-073 on mitochondrial priming (Δ% priming) and apoptotic cell death, human lung cancer A549 cells were treated with increasing concentrations of BKA-073 (0, 0.25, 0.5, 0.75, 1.0 μM) and then analyzed for dynamic BH3 profiling (DBP) at 16 hours and apoptotic cell death at 72 hours. DBP is a functional assay that can measure early changes in net pro-apoptotic signaling ("priming") at mitochondria induced by chemotherapeutic or targeted drugs in cancer cells. Priming is an indicator of how close cells are to the apoptotic threshold. Results showed that BKA-073 induced mitochondrial priming and apoptosis in a dose-dependent manner.

[0103] A panel of NSCLC and SCLC cell lines was tested. BKA-073 potently induced mitochondrial priming and apoptosis in both NSCLC and SCLC cell lines expressing various levels of endogenous Bak (Figure 1B). NSCLC cell lines (A549, H157 and H1975) and SCLC cell lines (i.e. DMS53, DMS114, H209 and H526) expressing relatively high levels of Bak were more sensitive to BKA-073. In contrast, lung cancer cell lines expressing relatively low levels of endogenous Bak (NSCLC cell line: Calu-1, SCLC cell lines: H69, H128 and H146) were less sensitive to BKA-073. Thus, the sensitivity of BKA-073 induction of mitochondrial priming and apoptosis is relatively dependent on the Bak expression level. In addition to lung cancer cell lines, the efficacy of BKA-073 was evaluated in other types of cancer cell lines, including breast cancer (MDA-MB-231 and MCF7), colon cancer (HCT-116), lymphoma (Ramos), multiple myeloma (OPM-1), pancreatic cancer (PANC-1) and osteosarcoma (U2OS) cell lines. The results revealed that BKA-073 potently induced mitochondrial priming and apoptotic cell death in various types of cancer cell lines as well (Figure 1C), suggesting that BKA-073 is effective against various types of cancer.

[0104] BKA-073 directly binds to Bak protein and induces Bak oligomerization, resulting in Cyt c release. To confirm the binding of BKA-073 to Bak, a competitive fluorescence polarization (FP) assay was performed using purified human Bak protein, fluorescent Bak BH3 domain peptide and BKA-073. BKA-073 directly bound to human Bak protein with high binding affinity (Ki: 72.3 ± 5.96 nM). Specifically, BKA-073 showed extremely low binding affinity to other Bcl2 family members (Figure 2A), indicating that BKA-073 selectively binds to Bak. There are multiple amino acid differences in the BH3 domain between Bak and other Bcl2 family members. BKA-073 appears to bind only to Bak and not to other Bcl2 family members (Bax, Bcl2, Bcl-XL, Bcl-w and Mcl-1).

[0105] Isothermal titration calorimetry (ITC) was also used to measure Bak / BKA-073 binding. ITC is a direct, label- and immobilization-free technique to measure binding affinity between interacting proteins and small molecule ligands, and can be used to analyze binding constant (Kd) values ​​in the millimolar and nanomolar ranges. ITC experiments were performed using an auto-iTC200 instrument to evaluate BKA-073 / Bak binding. The results show that BKA-073 bound directly to human Bak protein with a binding affinity in the nanomolar range (Kd = 88.62 ± 5.73 nM) (Figure 2B). In contrast, BKA-073 did not bind to the BH3-deleted human Bak mutant protein (ΔBH3) in the ITC assay, suggesting that the BH3 domain is involved in Bak interacting with BKA-073.

[0106] In addition to human Bak / BKA-073 binding, mouse Bak / BKA-073 binding was measured using ITC. BKA-073 also bound directly to mouse Bak protein with good binding affinity (Kd=93.37±7.91 nM). These experiments indicate that BKA-073 can bind to both human and mouse Bak proteins.

[0107] One step in the apoptotic process is the oligomerization of Bak. To assess whether BKA-073 affects the ability of Bak to form oligomers in mitochondrial membranes, cross-linking studies with bis(maleimido)hexane (BMH) were performed. Treatment of A549 cells with BKA-073 (1 μM) promoted the formation of Bak dimers and trimers. The molecular size of these adducts was estimated to be multiples of approximately 28 kDa, suggesting the formation of Bak homo-oligomers in A549 cells. These findings indicate that BKA-073 can activate Bak by oligomerization in mitochondria. The formation of Bak oligomers in mitochondria promotes the release of cytochrome c (Cyt c) to induce apoptosis. Experiments show that BKA-073-induced Bak oligomerization promoted Cyt c release from mitochondria in A549 cells.

[0108] BKA-073 potently suppresses NSCLC xenografts by inducing Bak-dependent apoptosis To test the efficacy of BKA-073 in vivo, mice bearing lung cancer xenografts derived from A549 cells were treated intraperitoneally for 28 days with increasing doses of BKA-073 (0, 5, 10, 15 mg / kg / day). BKA-073 potently inhibited lung cancer growth in a dose-dependent manner (Figure 3). To evaluate whether BKA-073-induced inhibition of tumor growth occurs through Bak activation and apoptosis in vivo, representative samples from harvested tumor tissues were analyzed by crosslinking with BMH for Bak oligomerization or by immunohistochemistry (IHC) for active caspase 3. Dose-dependent Bak oligomerization and apoptosis were observed in tumor tissues after BKA-073 treatment. Importantly, doses of 5 to 15 mg / kg / day not only potently inhibited tumor growth but were also well tolerated without significant toxicity to mice. Doses of 5 mg / kg to 15 mg / kg provide the optimal therapeutic index for BKA-073 in in vivo experiments involving lung cancer xenografts.

[0109] BKA-073 demonstrates potent antitumor activity against SCLC in xenograft and PDX models To further evaluate the antitumor activity of BKA-073 against SCLC in vivo, mice implanted with SCLC xenografts derived from the DMS114 cell line or patient-derived xenografts (PDXs) from two refractory SCLC patients (TKO-2 and TKO-5) were treated intraperitoneally with BKA-073 (15 mg / kg / d) for 2–4 weeks. BKA-073 potently suppressed tumor growth of DMS114 xenografts and SCLC PDXs caused by induction of apoptosis (Figure 4). These findings indicate that BKA-073 may be potentially effective in SCLC patients.

[0110] BKA-073 inhibits proliferation and extends survival of mutant KRAS-driven lung cancer in genetically engineered mouse models (GEMMs) KRAS is a commonly mutated oncogene, but there is no effective targeted therapy for KRAS mutant cancers. Interestingly, expression of exogenous constitutively active KRAS(G12D) mutant in H1944 cells with wild-type KRAS background significantly enhanced Bak expression. Since BKA-073 can induce apoptosis by activating Bak via promoting oligomerization in vitro and in vivo, experiments were performed to confirm whether BKA-073 is effective in treating mutant KRAS cancers.

[0111] To evaluate the efficacy of BKA-073 in lung cancer driven by KRAS mutations, we generated and bred lox-stop-lox (LSL)-KRAS G12D LKB1fl / fl (i.e., KL) mice. These mice carry a KRAS G12D LSL knock-in allele and a floxed allele of LKB1 (LKB1fl / fl). Primary lung adenocarcinomas were bred using 5 × 10 6After intranasal administration of pfu adenovirus, Bak expression was detectable as early as 6 weeks. Increased Bak expression was observed in tumor tissues from KL mice compared with adjacent normal lung tissue in representative sections from each lung lobe in each mouse. Results showed that treatment of KL mice with BKA-073 significantly reduced lung tumor burden and growth via apoptosis. Treatment with BKA-073 significantly extended survival of KL mice, providing a strong rationale for using the Bak agonist BKA-073 to treat mutant KRAS-driven lung cancer.

[0112] To further evaluate the potential of BKA-073 as a treatment for mutant KRAS-induced lung cancer, BKA-073 (15 mg / kg / day) or vehicle was administered intraperitoneally to KL mice starting 6 weeks after AdeCre delivery. After 48 days of treatment, KL mice were euthanized by carbon dioxide asphyxiation. Tumor-bearing lung and normal lung tissue were harvested for further analysis. To quantify the tumor burden and tumor multiplicity of mice, HE-stained lungs were imaged with morphometry software to quantify the surface area composed of tumor as opposed to normal tissue compared to the control group. Calculations up to 48 days before euthanasia showed that 4 of 6 mice died in the control group, whereas 2 of 6 mice died in the BKA-073-treated group (p<0.01) (Figure 5).

[0113] Bak accumulates in radioresistant lung cancer cells and BKA-073 reverses radioresistance in vitro and in vivo To further investigate whether Bak contributes to radioresistance, three lung cancer cell lines (i.e., A549-IRR, H358-IRR and H460-IRR) with resistance to ionizing radiation were established. Elevated Bak levels were observed in A549-IRR, H358-IRR and H460-IRR cells compared with parental A549 (A549-P), H358 (H358-P) and H460 (H460-P) cells. A549-IRR, H358-IRR and H460-IRR cells grew well under cell culture conditions, indicating that the Bak molecule is in an inactive form under normal growth conditions. A549-P, H385-P and H460-P cells remained sensitive to IR, whereas A549-IRR, H358-IRR and H460-IRR became insensitive to IR. Both the parental and radioresistant cell lines were sensitive to BKA-073, suggesting that BKA-073 is effective in radioresistant cells.

[0114] To further test this in vivo, NSCLC xenografts derived from A549-P and A549-IRR cell lines were treated with IR (2 Gy / exposure, every other day, total of 5 times) or BKA-073 (15 mg / kg / day) for 4 weeks. Lung cancer xenografts derived from A549-IRR cells were resistant to IR treatment, whereas xenografts derived from A549-P were sensitive to IR treatment. BKA-073 suppressed xenografts derived from A549-P or A549-IRR cells, indicating that BKA-073 is also effective against radiation-resistant lung cancer xenografts.

[0115] Combination of BKA-073 and the Bcl2 inhibitor venetoclax (ABT-199) synergistically inhibits lung cancer in vitro and in vivo To test whether direct activation of Bak proapoptotic activity combined with inhibition of Bcl2 antiapoptotic function could achieve synergistic effects in lung cancer treatment, SCLC (DMS53) and NSCLC (H460) cell lines expressing endogenous Bcl2 and Bak were treated with a combination of venetoclax and BKA-073 for 16 and 72 h, after which dynamic BH3 profiling and apoptosis were analyzed, respectively. The combination of BKA-073 and venetoclax demonstrated strong synergy in inducing mitochondrial priming and apoptosis in both SCLC and NSCLC lines. DMS53 cell-derived SCLC xenografts and H460 cell-derived NSCLC xenografts were treated with BKA-073 (10 mg / kg / day) intraperitoneally, venetoclax (60 mg / kg / day) orally, or the combination for 4 weeks. The results showed that combined treatment with BKA-073 and venetoclax synergistically inhibited both SCL and NSCLC in vivo (Figure 6).

[0116] Higher levels of Bak in tumor tissues correlate with poor prognosis of NSCLC patients Higher levels of endogenous Bak expression were observed in various human lung cancer cell lines, which did not induce apoptosis in cell culture medium without treatment, indicating that the Bak protein is in an inactive form under normal growth conditions. To further test whether Bak is upregulated in tumor tissues from NSCLC patients, Bak expression was analyzed by IHC staining with Bak antibody in samples from 208 NSCLC patients. Formalin-fixed, paraffin-embedded human tissue samples were obtained. Tissue microarrays (TMAs) were constructed using replicate cores of tumor and adjacent normal lung. Semiquantitative evaluation of IHC staining of Bak was performed using an "immunoscore" based on both the percentage of stained cells and the staining intensity, as described. Bak protein expression was significantly higher in tumor tissues compared to adjacent normal lung tissues. Importantly, increased Bak expression in tumor tissues correlated with poor prognosis of NSCLC patients (Figure 7), indicating that Bak is a potential prognostic biomarker for NSCLC. These experiments indicate that a Bak activator (BKA-073) is an effective strategy to improve outcomes for patients with NSCLC and other cancers.

Claims

A pharmaceutical composition for treating cancer, comprising a Bak activator.

2. The pharmaceutical composition according to claim 1, wherein the Bak activator is 1-((2-((2-methoxyacridin-9-yl)amino)ethyl)amino)propan-2-ol (BKA-073), its derivative, ester or salt.

3. The Bak activator is of formula I or II: 【Chemical Formula 1】 [wherein Q is O or S; U is N or CH; R 1 、 R 2 、 R 3 、 R 4 、 R 5 、 R 6 、 R 7 、 R 8 、 R 9 and R 10 are each individually and independently hydrogen, alkyl, halogen, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkanoyl, alkylthio, alkylamino, aminoalkyl, (alkyl) 2 amino, phosphate, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclic, aryl or heterocyclic, where R 1 、 R 2 、 R 3 、 R 4 、 R 5 、 R 6 、 R 7 、 R 8 、 R 9 and R 10 may be substituted with one or more identical or different R 11 ; R 11 is alkyl, halogen, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkanoyl, alkylthio, alkylamino, phosphate, aminoalkyl, (alkyl) 2 amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclic, aryl or heterocyclic, where R 11 may be substituted with one or more identical or different R 12 ; R 12 is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, 2-methoxyethoxy, 2-hydroxyethoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclic, aryl or heterocyclic] The pharmaceutical composition according to claim 1, which is a compound represented by, its ester or salt.

4. R 1 The pharmaceutical composition according to claim 3, wherein R is hydrogen.

5. R 2 The pharmaceutical composition according to claim 3, wherein R is alkyl.

6. R 3 The pharmaceutical composition according to claim 3, wherein R is hydrogen.

7. R 4 The pharmaceutical composition according to claim 3, wherein R is hydrogen.

8. R 5 The pharmaceutical composition according to claim 3, wherein R is alkyl.

9. R 6 、 R 7 、 R 8 、 R 9 and R 10 are hydrogen, the pharmaceutical composition according to claim 3.

10. The pharmaceutical composition according to claim 3, wherein Q is O.

11. The pharmaceutical composition according to claim 3, wherein U is NH.

12. The pharmaceutical composition according to claim 1, wherein the subject is a human.

13. The pharmaceutical composition according to claim 1, wherein the subject is diagnosed with non-small cell lung cancer.

14. The pharmaceutical composition according to claim 1, wherein the Bak activator is administered in combination with a further chemotherapeutic agent.

15. The pharmaceutical composition according to claim 14, wherein the chemotherapeutic agent is a Bcl-2 inhibitor.

16. The pharmaceutical composition according to claim 15, wherein the Bcl-2 inhibitor is venetoclax, navitoclax, obatoclax, subtoclax.

17. The pharmaceutical composition according to claim 1, further comprising a pharmaceutically acceptable additive.

18. The pharmaceutical composition according to claim 17, which is in the form of a pill, capsule or tablet.

19. The pharmaceutical composition according to claim 17, which is in the form of an isotonic or non-isotonic pH buffered aqueous solution.

20. The pharmaceutical composition according to claim 17, wherein the pharmaceutically acceptable additive is selected from monosaccharides, disaccharides, sucrose, lactose, glucose, mannitol, sorbitol, polysaccharides, starch, cellulose, crystalline cellulose, cellulose ether, hydroxypropyl cellulose (HPC), xylitol, maltitol, gelatin, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), hydroxypropylmethylcellulose (HPMC), crosslinked sodium carboxymethyl cellulose, dicalcium phosphate, calcium carbonate, stearic acid, magnesium stearate, talc, magnesium carbonate, silica, vitamin A, vitamin E, vitamin C, retinyl palmitate, cerium, cysteine, methionine, citric acid and sodium citrate, methylparaben, propylparaben, and combinations thereof.

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