Combination therapy and uses thereof

Abstract

The present disclosure relates to a combination of a BH-3 mimetic and a STING agonist and the use of the combination in combination therapy for the treatment of cancer.

Description

[0001] This application claims priority to Australian provisional patent applications 2023902434 (filed on 1 August 2023) and 2023903793 (filed on 24 October 2023), the entire contents of which are incorporated herein by reference. Technical Field

[0002] This disclosure relates to combination therapies and their use in medicine. More specifically, this disclosure relates to therapies comprising a combination of a BH3 mimic and a STING agonist suitable for the treatment and management of proliferative diseases such as cancer, and medical treatments using said therapies. Background Technology

[0003] Over the past decade, survival rates for many cancers have improved significantly due to advancements in molecular profiling and novel treatments for these malignancies. However, TP53 (also known as TRP53 in mice or commonly referred to as p53) mutations remain a major clinical challenge in cancer treatment. TP53 is the most frequently mutated gene across all cancer types, but TP53-mutant cancers are particularly resistant to cytotoxic drugs that rely on causing DNA damage to trigger cancer cell death. TP53-mutant cancer cells often also exhibit defects in a wide range of pathways, including metabolism, genomic stability, and autophagy, which reduces their sensitivity to a broad spectrum of anticancer agents. For hematologic malignancies such as lymphoma and leukemia, a subgroup of patients carrying TP53 mutations in their malignant cells is generally considered to be at adverse risk and have poor survival outcomes. TP53 mutations are present in 5%–15% of acute myeloid leukemia (AML) (up to 25% in elderly patients), 25% of non-Hodgkin lymphomas (NHL), and up to 60% of natural killer / T (NKT) cell lymphoma cases in some populations. Therefore, effective treatments for cancers with TP53 mutations, especially hematologic malignancies with TP53 mutations, remain an urgent but underresolved clinical need.

[0004] TP53 is activated in response to oncogene activation, DNA-damaging cytotoxic drugs, and other stressors such as gamma radiation. In these situations, it transcriptionally upregulates the expression of genes encoding pro-apoptotic BH3-only proteins (e.g., NOXA, PUMA, BIM), thereby initiating the intrinsic apoptosis pathway. Intrinsic apoptosis signaling is influenced by the balance between pro-apoptotic and anti-apoptotic members of the BCL-2 family. Pro-apoptotic BH3-only proteins bind to and block pro-survival proteins (e.g., MCL-1, BCL-2, BCL-XL), thereby promoting the activation of the pro-apoptotic effectors BAK and BAX. Activated BAK and BAX lead to mitochondrial outer membrane permeability (MOMP), which releases apoptosis-inducing factors into the cytoplasm, catalyzing the activation of the caspase cascade and cell destruction.

[0005] BH3 mimics are small molecule inhibitors that directly bind to and inhibit selected pro-survival members of the BCL-2 family. The identification of the functions of different BCL-2 family members, coupled with a deepening understanding of the structural interactions between pro-apoptotic and pro-survival family members, has given rise to the concept of killing cancer cells by targeting pro-survival members with small molecules that mimic the function of BH3-only proteins (now known as BH3 mimics).

[0006] The development of BH3 mimics is considered a major milestone toward developing a TP53-agnostic cancer therapy, as its use would allow for downstream activation of apoptosis. Unfortunately, although patients may initially respond well to treatment with BH3 mimics, recent clinical data indicate that most patients will relapse. A number of mechanisms enabling the spread of resistance to BH3 mimics have been identified, perhaps most surprisingly, that functional TP53 is essential for achieving maximum BH3 mimic-induced apoptosis in leukemia cells. While the underlying mechanisms of this TP53-mediated response to BH3 mimics are not yet fully understood, recent evidence suggests that patients with TP53-deficient hematologic malignancies, such as chronic lymphocytic leukemia (CLL) or AML, have poorer outcomes after treatment with the BCL-2-specific BH3 mimic venetoclax.

[0007] Therefore, cancer treatments that address one or more of the above problems, or at least provide a useful alternative, are desirable. In particular, improved methods that treat cancer or inhibit its progression by killing malignant cells regardless of their TP53 state (mutant or wild-type) are desirable.

[0008] The invention described herein provides a novel combination therapy that overcomes the TP53 mutation-driven barrier to successful treatment of other vulnerable cancers by BH3 mimics and possibly other anticancer agents. Specifically, the disclosure describes a combination of a BH3 mimic with a STING agonist that synergistically enhances the killing of cancer cells in a TP53-independent manner. This combination provides a novel treatment regimen that targets both wild-type and mutant TP53 cancer cells, thereby potentially avoiding the development of therapy-resistant TP53 mutant cell subsets and / or TP53 mutant cell subsets that can induce disease relapse. Therefore, the combination described herein is suitable for combating refractory or relapsed cancers, including those previously treated with TP53-sensitive compounds (such as DNA damage-inducing chemotherapeutic agents or BH3 mimics) and induction therapies for newly diagnosed diseases.

[0009] References to patent documents or other matters given in this document as prior art should not be construed as an admission that the documents or matters are known or that the information contained therein is part of common general knowledge from the priority date of any claim.

[0010] definition C 3-6 Cycloalkyl: As used herein, the term "C" 3-6 "Cycloalkyl" refers to the monovalent portion obtained by removing hydrogen atoms from the carbon atoms of a saturated cyclic hydrocarbon compound having 3 to 6 carbon atoms. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl (C3), cyclobutyl (C4), cyclopentyl (C5), and cyclohexyl (C6).

[0011] C 3-7 Heterocyclic group: As used herein, the term "C" 3-7 "Heterocyclic group" refers to a monovalent moiety obtained by removing hydrogen atoms from the ring atoms of a monocyclic heterocyclic compound, said moiety having 3 to 7 ring atoms; wherein 1 to 2 atoms are heteroatoms selected from oxygen, sulfur, or nitrogen. C 3-7 Heterocyclic groups can be non-aromatic ring systems or aromatic ring systems. Aromatic C 3-7 Heterocyclic groups can be called C 3-7 Mixed aromatic compounds.

[0012] In this context, prefixes (e.g., C) 3-7 The prefix () indicates the number or range of ring atoms, whether carbon or heteroatoms. Therefore, in some cases, the prefix can be interchanged with alternative prefixes that define the number of ring member atoms, such as the prefix "C". 3-7"Can be interchanged with the prefix "3 to 7". In some embodiments, C in the compounds of the present invention 3-7 The heterocyclic moiety can be a C3, C4, C5, C6, or C7 heterocyclic group, or any combination of these different-sized ring / ring systems, such as C 3-6 C 4-7 Or C 5-6 Heterocyclic group.

[0013] C 3-7 Examples of heterocyclic groups include, but are not limited to, those derived from the following heterocyclic groups: N1: Aziridine (C3), aziridine (C4), pyrrolidine (tetrahydropyrrole) (C5), pyrrololine (e.g., 3-pyrrololine, 2,5-dihydropyrrole) (C5), 2H-pyrrole or 3H-pyrrole (isopyrrole, isazole) (C5), pyrrole (C5), piperidine (C6), dihydropyridine (C6), tetrahydropyridine (C6), pyridine (C6), aziridine heptatriene (C7), aziridine heptatriene (C7); N2: diazirine (C3), diazacyclobutane (C4), imidazoline (C5), pyrazolidine (diazolidine) (C5), imidazoline (C5), pyrazolidine (dihydropyrazole) (C5), imidazolium (C5), pyrazole (C5), piperazine (C6), pyrazine (C6), pyrimidine (C6), pyridazine (C6), diazacycloheptatriene (C7), diazacycloheptatriene (C7); O1: Oxybutane (C4), tetrahydrofuran (C5); oxane (C6); O2: dioxetane (C4), dioxopentane (C5); dioxane (C6), dioxetene (C5); N1O1: Tetrahydrooxazole (C5), Dihydrooxazole (C5), Tetrahydroisoxazole (C5), Dihydroisoxazole (C5), Isoxazole (C5), Oxazole (C5), Morpholine (C6), Tetrahydrooxazine (C6), Dihydrooxazine (C6), Oxazine (C6); S1: thiirane (C3), thiazole butane (C4), thiazole pentane (tetrahydrothiophene) (C5), thiophene (C5), thiane (tetrahydrothiaran) (C6), thiepane (C7); N1S1: Thiazoline (C5), Thiazolidine (C5), Thiazole (C5), Isothiazole (C5), Thiomorpholine (C6), Thiazide (C6); O1S1: oxathiolidene (C5), isoxthiolidine (C5), oxathiole (C5), isoxathiolane (C5) and oxathiane (thiamane) (C6); N2O: Oxadiazole (C5); N2S: Thiadiazole (C5).

[0014] C 1-4 Alkyl: As used herein, the term "C" 1-4 "Alkyl" refers to the monovalent portion obtained by removing hydrogen atoms from the carbon atoms of a saturated hydrocarbon compound having one to four carbon atoms.

[0015] Examples of saturated alkyl groups include, but are not limited to, Me: methyl (C1), Et: ethyl (C2), Pr: propyl (C3), and Bu: butyl (C4).

[0016] Examples of saturated straight-chain alkyl groups include, but are not limited to, methyl (C1), ethyl (C2), nPr: n-propyl (C3), and nBu: n-butyl (C4).

[0017] Examples of saturated branched alkyl groups include, but are not limited to, iPr: isopropyl (C3, -C(CH3)2), iBu: isobutyl (C4), sBu: sec-butyl (C4), and tBu: tert-butyl (C4).

[0018] C2-4 alkenyl: As used herein, the term “C2-4 alkenyl” refers to an alkyl group having 2 to 4 carbon atoms and having one or more carbon-carbon double bonds.

[0019] Examples of unsaturated alkenyl groups include, but are not limited to, methine (vinyl, -CH=CH2), 1-propenyl (-CH=CH-CH3), 2-propenyl (allyl, -CH-CH=CH2), isopropenyl (1-methylvinyl, -C(CH3)=CH2) and butenyl (C4).

[0020] C 1-4 Fluoroalkyl: As used herein, the term "C" 1-4 "Fluoroalkyl" refers to a C that has been substituted with one or more fluorine atoms. 1-4 alkyl.

[0021] Alkoxy group: -OR, where R is C as defined above. 1-4 Alkyl group. It can be represented as -OC. 1-4Alkyl groups. Examples of alkoxy groups include, but are not limited to, methoxy (OMe, C1), ethoxy (OEt, C2), propoxy (C3), and butoxy (C4).

[0022] Alkyl carbamoyl group: -NHC(=O)OR, where R is C as defined above. 1-4 Alkyl. Examples of alkyl carbamoyl groups include, but are not limited to, -N(H)C(=O)OCH3, -N(H)C(=O)OCH2CH3 and -N(H)C(=O)OC(CH3)3.

[0023] Alkyl carbamoyl ester: -OC(=O)NRR', where R and R' are independently selected from H and C as defined above. 1-4 Alkyl groups. Examples of alkyl carbamoyl ester groups include, but are not limited to, -OC(=O)N(CH3)2 and -OC(=O)N(H)CH3.

[0024] Alkyl carboxyl esters: -OC(=O)OR, where R is C as defined above. 1-4 Alkyl groups. Examples of alkyl carboxyl ester groups include, but are not limited to, -OC(=O)OCH3, -OC(=O)OCH2CH3, -OC(=O)OC(CH3)3 and -OC(=O)OCH(CH3)2.

[0025] Amino group: -N(R)R', where R and R' are independently selected from H and C as defined above. 1-4 Alkyl groups. Examples of amino groups include, but are not limited to, -NH2, -N(H)CH3, -N(H)C(CH3)2, -N(CH3)2, and -N(CH2CH3)2.

[0026] Amide group (carbamoyl, carbamyl, aminocarbonyl, formamide, aminoacyl): -C(=O)N(R)R', where R and R' are independently selected from H and C as defined above. 1-4 Alkyl groups. Examples of amide groups include, but are not limited to, C(=O)NH2, -C(=O)N(H)CH3, -C(=O)N(CH3)2, -C(=O)N(H)CH2CH3 and -C(=O)N(CH2CH3)2.

[0027] Acylamide group: -N(R)C(=O)R', where R and R' are independently selected from H and C as defined above. 1-4 Alkyl groups. Examples of acyl amide groups include, but are not limited to, -N(H)C(=O)CH2CH3, -N(H)C(=O)CH3 and -N(CH3)C(=O)CH3.

[0028] Phenyl: -C6H5, wherein the phenyl group itself may optionally be converted by one or more C6H5 groups. 1-4 Alkyl, one or more C 1-4 fluoroalkyl, one or more C 1-4 The alkoxy group, one or more halogenated substituents, and one or more cyano substituents are substituted.

[0029] Benzyl: -CH2-phenyl, where phenyl is as defined above.

[0030] Ester (carboxylate, carboxylic acid ester, oxycarbonyl): -C(=O)OR, where R is an ester substituent, for example, C 1-4 Alkyl, C 3-7 Heterocyclic group or phenyl group as defined above, preferably C 1-4 Alkyl groups. Examples of ester groups include, but are not limited to, -C(=O)OCH3, -C(=O)OCH2CH3, -C(=O)OC(CH3)3 and -C(=O)OPh.

[0031] Acyloxy group (reverse ester): -OC(=O)R, where R is an acyloxy substituent, such as C 1-4 Alkyl, C 3-7 Heterocyclic group or phenyl group as defined above, preferably C 1-4 Alkyl groups. Examples of acyl groups include, but are not limited to, -OC(=O)CH3 (acetoxy), -OC(=O)CH2CH3, -OC(=O)C(CH3)3, and -OC(=O)Ph. Further examples of acyl groups include, but are not limited to, methyl esters (C1), ethyl esters (C2), propyl esters (C3), and butyl esters (C4).

[0032] Naturally occurring amino acids: As used herein, the term "naturally occurring amino acid" refers to the monovalent portion obtained by removing a hydrogen atom from the carboxyl or amino group of one of the amino acid compounds commonly found in nature (e.g., alanine, arginine, asparagine, aspartic acid, cysteine, glycine, glutamine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine). Amino acids are particularly selected from isoleucine, leucine, and valine, with valine being the most prominent.

[0033] In each of these groups, the carbon atom bonded to both the carboxyl and amino groups is called the α-carbon, and the carboxyl and amino groups it is bonded to are α-carboxyl and α-amino, respectively. Naturally occurring amino acids may optionally be substituted by a protecting group on the α-amino group or by any other amino group on the α-amino group. Protecting groups include, but are not limited to, acetyl, methyl, fluorenylmethoxycarbonyl (Fmoc), carboxybenzyl (Cbz; benzyloxycarbonyl), phthalimide, and tert-butyl carbamate (boc) groups.

[0034] Phosphonates: -P(O)(OR)OR', where R and R' are independently selected from C as defined above. 1-4 Alkyl groups. Examples of phosphonates include, but are not limited to, -P(O)(OEt)2.

[0035] Cyano group: -C≡N.

[0036] Neopentyloxymethyl: a group having the following formula .

[0037] Tetrazolyl: A group having the following formula or .

[0038] Oxo-thiadiazole group: A five-membered heterocyclic group having two nitrogen ring atoms, a sulfur ring atom, and an oxygen substituent. Examples of oxo-thiadiazole groups include:

[0039] Thio-thiadiazole group: A 5-membered heterocyclic group having two nitrogen ring atoms, a thio ring atom, and a thiocarbonyl substituent. Examples of thio-thiadiazole groups include:

[0040] Thio-oxadiazole group: A 5-membered heterocyclic group having two nitrogen ring atoms, an oxygen ring atom, and a thiocarbonyl substituent. Examples of thio-oxadiazole groups include:

[0041] Hydroxy-thiadiazole group: A five-membered heterocyclic group having two nitrogen ring atoms, a sulfur ring atom, and a hydroxyl substituent. These groups can exhibit tautomerism with their corresponding oxo-thiadiazole groups under certain conditions. Examples of hydroxy-thiadiazole groups include:

[0042] Hydroxy-oxadiazole group: A five-membered heterocyclic group having two nitrogen ring atoms, an oxygen ring atom, and a hydroxyl substituent. These groups can exhibit tautomerism with their corresponding oxo-oxadiazole groups under certain conditions. Examples of hydroxy-oxadiazole groups include:

[0043] Mercapto-thiadiazole group: A five-membered heterocyclic group having two nitrogen ring atoms, a sulfur ring atom, and a mercapto substituent. These groups can exhibit tautomerism with their corresponding thio-thiadiazole groups under certain conditions. Examples of mercapto-thiadiazole groups include:

[0044] Thio-oxadiazole group: A five-membered heterocyclic group having two nitrogen ring atoms, an oxygen ring atom, and a thiol substituent. These groups can exhibit tautomerism with their corresponding thio-oxadiazole groups under certain conditions. Examples of thio-oxadiazole groups include:

[0045] Hydroxy-oxazolyl: A five-membered heterocyclic group having a nitrogen ring atom, an oxygen ring atom, and a hydroxyl substituent. Examples of hydroxy-oxazolyl groups include:

[0046] In some embodiments, the hydroxy-oxazolyl group may be 3-hydroxy-1,2-oxazol-5-yl.

[0047] Thio-oxazolyl: A 5-membered heterocyclic group having a nitrogen ring atom, an oxygen ring atom, and a thiol substituent. Examples of thio-oxazolyl groups include:

[0048] In some embodiments, the hydroxy-oxazolyl group may be 3-mercapto-1,2-oxazol-5-yl.

[0049] Hydroxythiazolyl: A five-membered heterocyclic group having a nitrogen ring atom, a sulfur ring atom, and a hydroxyl substituent. Examples of hydroxythiazolyl groups include:

[0050] In some embodiments, the hydroxy-thiazolyl group may be 3-hydroxy-1,2-thiazolyl-5-yl.

[0051] Mercapto-thiazolyl: A 5-membered heterocyclic group having a nitrogen ring atom, a sulfur ring atom, and a mercapto substituent. Examples of mercapto-thiazolyl groups include:

[0052] In some embodiments, the hydroxy-oxazolyl group may be 3-mercapto-1,2-thiazolyl-5-yl.

[0053] Hydroxydiazole group: A 5-membered heterocyclic group having two nitrogen ring atoms and a hydroxyl substituent. Examples of hydroxydiazole groups include:

[0054] 2H-triazol-4-yl: A group having the following formula: .

[0055] Unless the context otherwise requires, where the terms “comprise,” “comprises,” and “comprising” are used in this specification (including the claims), these shall be construed as designating the stated feature, integer, step, or component, but do not exclude the presence of one or more other features, integers, steps, or components, or groups thereof.

[0056] It should be further understood that terms such as “comprise” or variations thereof, such as “comprises” or “comprising”, inherently include (but are not limited to) inventive versions that exclude other elements directly related to the present invention within their scope. Therefore, terms such as “composed of” or “generally composed of” can be replaced by terms such as “comprise,” “comprises,” or “comprising”, which serve to limit the scope of the invention to the specifically mentioned elements. It is noteworthy that, where the invention is explicitly intended to be considered in an exhaustive manner, such limitations should be regarded as relating only to the inventive concepts disclosed herein, and other features falling outside the scope of the inventive concepts may be added. Such features or elements may include, but are not limited to, excipients, formulations, additives, diluents, packaging, adjuvants, and parallel features that should not be excluded by terms such as “composed of” or “generally composed of”. Summary of the Invention

[0057] According to a first aspect of the invention, a method for treating or inhibiting the progression of cancer in a subject is provided, the method comprising co-administering to the subject: a BH3 mimicry drug; and a STING agonist. In one embodiment, a method for treating or inhibiting the progression of cancer in a subject is provided, the method comprising co-administering to the subject: a BH3 mimicry drug; and a STING agonist, wherein the cancer comprises cells expressing the STING protein. In one embodiment, a method for treating or inhibiting the progression of cancer in a subject is provided, the method comprising co-administering to the subject a synergistic combination of: a BH3 mimicry drug; and a STING agonist. In one embodiment, a method for treating or inhibiting the progression of cancer in a subject is provided, the method comprising co-administering to the subject: a BH3 mimicry drug; and a STING agonist, wherein the cancer comprises cells expressing the STING protein. In one embodiment, a method for treating or inhibiting the progression of cancer in a subject is provided, the method comprising co-administering to the subject a synergistic combination of: a BH3 mimicry drug; and a STING agonist, wherein the cancer comprises cells expressing the STING protein.

[0058] According to a second aspect of the invention, a method for inducing an immune response to cancer in a subject is provided, the method comprising: co-administering a BH3 mimic drug and a STING agonist to the subject, thereby inducing an anti-cancer immune response in the subject. The method may include co-administering a synergistic combination of the following to the subject: a BH3 mimic drug; and a STING agonist. In one embodiment, a method for inducing an immune response to cancer in a subject is provided, the method comprising: co-administering a BH3 mimic drug and a STING agonist to the subject, thereby inducing an anti-cancer immune response in the subject, wherein the cancer comprises cells expressing the STING protein. The method may include administering a synergistic combination of the BH3 mimic drug and the STING agonist to the subject.

[0059] According to a third aspect of the invention, a method is provided for treating cancer in a subject or inhibiting its progression, the method comprising: identifying the subject as a candidate for a BH3 mimicry drug therapy; identifying the cancer as cells containing STING protein; and administering both a BH3 mimicry drug and a STING agonist to the subject, wherein the STING agonist and the BH3 mimicry drug are administered sequentially or simultaneously. The method may include administering a synergistic combination of the BH3 mimicry drug and the STING agonist to the subject.

[0060] According to a fourth aspect of the invention, a method is provided for treating or inhibiting the progression of cancer in a subject who has received BH3 mimicry therapy (or treating or inhibiting the progression of cancer while the subject is receiving BH3 mimicry therapy), the method comprising administering a STING agonist to the subject. In one embodiment, a method is provided for treating or inhibiting the progression of cancer in a subject who has received BH3 mimicry therapy (or treating or inhibiting the progression of cancer while the subject is receiving BH3 mimicry therapy), the method comprising administering a STING agonist to the subject, wherein the cancer comprises cells expressing the STING protein. In one embodiment, a method is provided for treating or inhibiting the progression of cancer in a subject who has received BH3 mimicry therapy (or treating or inhibiting the progression of cancer while the subject is receiving BH3 mimicry therapy), the method comprising administering a synergistic amount of a STING agonist to the subject, such that the STING agonist and the BH3 mimicry provide a synergistic combination. In one embodiment, a method is provided for treating or inhibiting the progression of cancer in a subject who has received BH3 mimicry therapy (or treating or inhibiting the progression of cancer while the subject is receiving BH3 mimicry therapy), the method comprising administering a STING agonist to the subject, wherein the cancer comprises cells expressing the STING protein. In another embodiment, a method is provided for treating or inhibiting the progression of cancer in a subject who has received BH3 mimicry therapy (or treating or inhibiting the progression of cancer while the subject is receiving BH3 mimicry therapy), the method comprising administering a synergistic amount of a STING agonist to the subject to provide a synergistic combination of the BH3 mimicry and the STING agonist, wherein the cancer comprises cells expressing the STING protein.

[0061] The following features may be used alone or in any suitable combination with any of the first to fourth aspects described above.

[0062] The cancer may comprise TP53 wild-type cells and TP53 mutant / deficient cells. The cancer may be refractory or resistant to BH3 mimicry monotherapy. Co-administration may comprise simultaneous administration of the BH3 mimicry and the STING agonist, or sequential administration of the BH3 mimicry and the STING agonist. The BH3 mimicry may be administered orally or intravenously. The STING agonist may be administered intravenously or intratumorally, typically intravenously.

[0063] The BH3 mimic and the STING agonist can each be administered at a therapeutically effective amount. The BH3 mimic can be selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845. The BH3 mimic can be selected from venetoclax, A-1331852, and S63845. In embodiments, the STING agonist can be selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. The STING agonist can be selected from MSA-2, diABZI STING agonist 1, and ADU-S100 / MIW815. In embodiments, the STING agonist can be any compound of formula (I) to formula (V).

[0064] In some embodiments, the STING agonist is a compound of formula (I): (I) in: W is either O or NH; R 1 Selected from: iH; ii.C 3-6 cycloalkyl; iii.C 3-7 The heterocyclic group, which is optionally substituted with a group selected from the following: methyl; and Esters; and iv. Straight or branched chain C 1-4 Alkyl groups, which are optionally substituted with groups selected from the following: Alkoxy; amino; amide group; Acyl amide group; Acyloxy group; Alkyl carboxyl esters; Alkyl carbamoyl; Alkyl carbamoyl ester; Phenyl; Phosphonates; C 3-7 Heterocyclic groups, optionally substituted with groups selected from methyl and oxo groups; and Naturally occurring amino acids, wherein the naturally occurring amino acids are optionally N-substituted with groups selected from methyl, acetyl, and boc; A 1 It is CR A Or N; A 2 It is CR B Or N; A 3 It is CR C Or N; A 4 It is CR D Or N; Where A 1 A 2 A 3 and A 4 No more than two of them can be N; R A R B R C and R D One or both of them (if present) are selected from H, F, Cl, Br, Me, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe and OH; R A R B R C and R D The remainder (if it exists) is H; X is O, NH, or CH2; R Y Selected from: (a) , (R YA ); Z 1 It is CR Z1 Or N; Z 2 It is CR Z2 Or N; Z 4 It is CR Z4 Or N; Z5 is CR Z5 Or N; Z 1 Z 2 Z 4 and Z 5 No more than two of them can be N; R Z1 R Z2 R Z4 and R Z5 One or both of (if present) are selected from H, F, Cl, Br, Me, OMe, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups; R Z1 RZ2 R Z4 and R Z5 The remainder (if it exists) is H; (b) , (R YB ); Where R 12 Selected from H, F, Cl, Br, OMe, cyano, and CF3; The condition is when A 1 When it is CF, A 2 A 3 and A 4 It is CH; Y is O or NH; R Y It is R YA Z 1 Z 2 Z 4 and Z 5 It is CH;R 1 Not Et; and When A 1 When it is CF, A 2 A 3 and A 4 It is CH; Y is NH; R Y It is R YA Z 1 and Z 5 It is CH, Z 2 and Z 4 One of them is CF, and Z 2 and Z 4 The other one is CH;R 1 It's not Et.

[0065] In some embodiments, the STING agonist is a compound of formula (II): (II) in: W 1 It is O or NH; R 2 Selected from: iH; ii.C 3-6 cycloalkyl; iii.C 3-7 The heterocyclic group, which is optionally substituted with a group selected from the following: methyl; and Esters; and iv. Straight or branched chain C 1-4 Alkyl groups, which are optionally substituted with groups selected from the following: Alkoxy; amino; amide group; Acyl amide group; Acyloxy group; Alkyl carboxyl esters; Alkyl carbamoyl; Alkyl carbamoyl ester; Phenyl; Phosphonates; C 3-7 Heterocyclic groups, optionally substituted with groups selected from methyl and oxo groups; and Naturally occurring amino acids, wherein the naturally occurring amino acids are optionally N-substituted with groups selected from methyl, acetyl, and boc; A 5 It is CR E Or N; A 6 It is CR F Or N; A 7 It is CR G Or N; A 8 It is CR H Or N; Where A 5 A 6 A 7 and A 8 No more than two of them can be N; R A R B R C and R D One or both of them (if present) are selected from H, F, Cl, Br, Me, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe and OH; R E R F R G and R H The remainder (if it exists) is H; R N1 It is H or Me; R C2 and R C3 One of them is C(=O)N H2 The other is selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups; R C1 and R C4Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups.

[0066] In some embodiments, the STING agonist is a compound of formula (III): (III): in: Y is (CH2)n, where n is 2 to 4; W 2 and W 3 Independently selected from OH and OR P , where R P Is it Me or Et? A 11 It is CR I Or N; A 12 It is CR J Or N; A 13 It is CR K Or N; A 14 It is CR L Or N; Where A 11 A 12 A 13 and A 14 No more than two of them can be N; R I R J R K and R L One or both of them (if present) are selected from H, F, Cl, Br, Me, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe and OH; R I R J R K and R L The remainder (if it exists) is H; A 21 It is CR IA Or N; A 22 It is CR JB Or N; A 23 It is CR KC Or N; A 24 It is CR LD Or N; Where A 21 A22 A 23 and A 24 No more than two of them can be N; R IA R JB R KC and R LD One or both of them (if present) are selected from H, F, Cl, Br, Me, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe and OH; R IA R JB R KC and R LD The remainder (if it exists) is H; R C5 R C6 and R C7 Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups; R C15 R C16 and R C17 Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups.

[0067] In some embodiments, the STING agonist is a compound of formula (IV): (IV): in: Y is (CH2) n Or -CH2-CH=CH-CH2-, where n is 2 to 4; R 1a and R 11a Independently select from the following groups: -C(=O)OH, -C(=O)OR P1 , Br, F, tetrazolium, oxo-oxadiazolium and (2H-triazol-4-yl), -S(=O)2OH, -P(=O)(OH)2, Br, F, tetrazolyl, oxo-oxadiazolyl, , (4H-triazol-3-yl), (2H-triazol-4-yl), oxo-thiadiazolyl, thio-thiadiazolyl, thio-oxadiazolyl, hydroxy-oxadiazolyl, hydroxy-thiadiazolyl, mercapto-oxadiazolyl, mercapto-thiadiazolyl, -C(CR)a R b R c (CR) x R y R z XH, hydroxy-oxazolyl, mercapto-oxazolyl, hydroxy-diazolyl, hydroxy-thiazolyl, mercapto-thiazolyl; R P1 Selected from methyl and ethyl; Each R a R b R c R x R y and R z Independently selected from H and F; X is selected from O and S; A 31 It is CR A1 Or N; A 32 It is CR B1 Or N; A 33 It is CR C1 Or N; A 34 It is CR D1 Or N; Where A 31 A 32 A 33 and A 34 No more than two of them can be N; R A1 R B1 R C1 and R D1 One, two, or three of (if present) are selected from H, F, Cl, Br, I, Me, Et, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe, and OH; R A1 R B1 R C1 and R D1 The remainder (if it exists) is H; A 41 It is CR A2 Or N; A 42 It is CR B2 Or N; A 43 It is CR C2 Or N; A 44 It is CR D2 Or N; Where A41 A 42 A 43 and A 44 No more than two of them can be N; R A2 R B2 R C2 and R D2 One, two, or three of (if present) are selected from H, F, Cl, Br, I, Me, Et, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe, and OH; R A2 R B2 R C2 and R D2 The remainder (if it exists) is H; R C1a R C3a and R C4a Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups; R C11a R C13a and R C14a Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups.

[0068] In some embodiments, the STING agonist is a compound according to formula (V): (V) in: Z is a 3-6 atom linker consisting of 1-6 -CH2- motifs and 0, 1 or 2 motifs independently selected from O, NH and -NHC(O)-; Y 1 It is H and Y 11 Is it H, or Y? 1 and Y 11 Together they form (CH2). n Or -CH2-CH=CH-CH2-, where n is 2 or 3; R 1 and R 11 Independently selected from -C(=O)OH, bioisosteres of carboxylic acids, Br, and F; A 1 It is CR A Or N; A 2 It is CRB Or N; A 4 It is CR D Or N; Where A 1 A 2 and A 4 No more than two of them can be N; R A R B and R D (If present) Independently selected from H, F, Cl, Br, I, Me, Et, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe, and OH; A 11 It is CR AA Or N; A 13 It is CR CC Or N; A 14 It is CR DD Or N; Where A 11 A 13 and A 14 No more than two of them can be N; R AA R CC and R DD (If present) Independently selected from H, F, Cl, Br, I, Me, Et, CF3, cyclopropyl, cyano, OMe, OEt, CH2OH, CH2OMe, and OH; R C1 R C3 and R C4 Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups; R C11 R C13 and R C14 Independently selected from H, Cl, F, Br, Me, OMe, OEt, cyano, CF3, CH2OH, CH2OMe, C 2-4 Alkenyl and C5 heterocyclic groups.

[0069] The compounds described herein, including those of formulas (I)-(V), may be provided in the form of pharmaceutically acceptable salts, solvates, prodrugs, isomers, tautomers, polymorphs, and / or N-oxides.

[0070] The BH3 mimic can be administered at a dose of about 50 mg / day to about 600 mg / day. The BH3 mimic can be administered at a dose equivalent to about 50 mg / day to about 600 mg / day of venetoclax. In any of the methods described herein, the STING agonist can be administered at any effective dose. The effective dose can be any amount that, in combination with the BH3 mimic, induces the desired physiological response. As is typically adjusted when determining the dosage of the drug component, the dose can be varied based on a variety of factors, including disease severity and subject characteristics (including height, weight, sex, medical history, etc.). In some embodiments, the method may comprise administering the STING agonist at a dose of about 10 µg / week to about 6,400 µg / week over a period of one to three doses every three weeks.

[0071] The cancer may be a blood cancer. The blood cancer may be selected from leukemias such as acute myeloid leukemia (AML); lymphomas such as T-cell lymphoma and multiple myeloma (MM); or any relapsed / refractory form of these diseases. The blood cancer may be selected from acute myeloid leukemia (AML), T-cell lymphoma, and multiple myeloma (MM), or any relapsed / refractory form of these diseases. The blood cancer may be selected from acute myeloid leukemia (AML), natural killer / T-cell lymphoma (NKTL), extranodal NK / T-cell lymphoma (ENKTL), and multiple myeloma (MM), or any relapsed / refractory form of these diseases. The cancer may be selected from the following leukemias: acute myeloid leukemia (AML), including promyelocytic leukemia, chronic myeloid leukemia (CML), and acute lymphoblastic leukemia (ALL), or any relapsed / refractory form of these diseases. The cancer may be a lymphoma. The cancer may be non-Hodgkin's lymphoma (NHL), such as that selected from adult T-cell lymphoma, lymphoblastic lymphoma, peripheral T-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma, natural killer / T-cell lymphoma (NKTL), extranodal NK / T-cell lymphoma (ENKTL), marginal zone lymphoma, Waldenstrom's macroglobulinemia, and mantle cell lymphoma, or a relapsed / refractory form of any of these diseases. The cancer may be a solid tumor. The solid tumor may be selected from small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, melanoma, breast cancer, ovarian cancer, neuroblastoma, prostate cancer, and colorectal cancer.

[0072] The subjects can be mammals, such as humans.

[0073] According to a fifth aspect of the invention, a combination for treating cancer or inhibiting its progression is provided, the combination comprising: a BH3 mimicry drug; and a STING agonist. In one embodiment, a combination for treating cancer or inhibiting its progression is provided, the combination comprising: a BH3 mimicry drug; and a STING agonist, wherein the cancer comprises cells expressing the STING protein. The combination may be a synergistic combination.

[0074] The following features may be used alone or in any suitable combination with the fifth aspect above.

[0075] The combination may be in the form of a pharmaceutical composition. The BH3 mimic and the STING agonist may be individual formulations. The BH3 mimic and the STING agonist may be administered simultaneously. The BH3 mimic and the STING agonist may be administered sequentially. The BH3 mimic may be selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845. The BH3 mimic may be selected from venetoclax, A-1331852, and S63845. In embodiments, the STING agonist may be selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. The STING agonist may be selected from MSA-2, diABZI STING agonist 1, and ADU-S100 / MIW815. In the embodiments, the STING agonist may be any compound of formula (I) to formula (V).

[0076] The cancer may contain cells expressing the STING protein. The cancer may contain TP53 wild-type cells and TP53 mutant cells. The cancer may be refractory or resistant to BH3 mimicry monotherapy.

[0077] According to a sixth aspect of the invention, there is provided the use of a BH3 mimicry and a STING agonist for preparing a medicament for treating cancer in a subject, wherein the BH3 mimicry and the STING agonist are administered co-administered. The co-administration may be sequential or simultaneous.

[0078] According to a seventh aspect of the invention, there is provided the use of a STING agonist for preparing a medicament for use in combination with a BH3 mimicry drug to treat cancer in a subject. The combination administration may be sequential or simultaneous.

[0079] According to an eighth aspect of the invention, there is provided the use of a BH3 mimicry for preparing a medicament for use in combination with a STING agonist to treat cancer in a subject. The combination administration may be sequential or simultaneous.

[0080] According to a ninth aspect of the present invention, a kit is provided which comprises, in a separate component: •BH3 mimicry drug •STING agonist • And optional instructions for use, for example, in any of the methods described herein, in combination of the BH3 mimicry and the STING agonist.

[0081] According to a tenth aspect of the present invention, a kit is provided which comprises, in a separate component: •BH3 mimicry drug • And instructions for use, for example, in combination with the STING agonist in any of the methods described herein.

[0082] According to an eleventh aspect of the present invention, a kit is provided which comprises, in a separate component: • Sting agonists; and • Instructions for use, for example, in combination of the STING agonist with a BH3 mimicry in any of the methods described herein. Attached Figure Description

[0083] This document will illustrate embodiments of the invention by way of example only, with reference to the accompanying drawings, in which: Figure 1 This study demonstrates the combination of BH3 mimicry drugs targeting MCL-1 or BCL-2 with STING agonist drugs to enhance the effects of cultured mice. Eµ-Myc Apoptosis of lymphoma cells. (A) Treatment of cultured cells with the MCL-1 inhibitor S63845 in combination with the STING agonist ADU-S100, MSA-2, or diABZI for 24 hours as a control of the syngeneic non-targeted sgRNA (NT). Trp53 KO and Bak / Bax Double KO mice Eµ-Myc Cell viability assay of lymphoma cell lines. (B) Isogenic NT control lymphoma cells Compared to That Trp53Cell competition assay of KO derivatives. Control (grey), S63845 (red), ADU-S100 (blue), or a combination of S63845 and ADU-S100 (purple). (C) Cell viability assay of mouse double-hit lymphoma (DHL) cell lines treated in culture for 24 hours with the BCL-2 inhibitor venetoc (10 nM) in combination with the STING agonist ADU-S100, MSA-2, or diABZI. (D, E) Transplanted syngeneic NT control (D) or Trp53 KO(E)AH15A mice Eµ-Myc Lymphoma cells were then treated with a mordant (black), diABZI (blue), S63845 (yellow), or diABZI with S63845 (pink). Rag1 Survival curves of mice. Boxes indicate when diABZI (blue) and S63845 (yellow) were administered.

[0084] Figure 2 This study demonstrates the combination of a BH3 mimic targeting BCL-XL with a STING agonist to enhance apoptosis in cultured human extranodal natural killer / T (ENKT) lymphoma cell lines. (A) Western blotting of STING protein expression in a human ENKT lymphoma cell line manifold. (B) [The text then abruptly shifts to a different topic:] ...for two human ENKT lymphoma cell lines (SNK6 (wild-type...)... TP53 ) and MEC04 (mutant) TP53(C) Western blot of cGAS / STING pathway activation. (D) Cell viability and proliferation data of SNK6 cells treated with STING agonists for 72 hours in culture (ADU-S100: 10 µg / mL, MSA-2: 34 µM, diABZI 1 µM). (E) Cell viability assay of three human ENKT lymphoma cell lines (SNK6, MEC04, SNT15) treated with a combination of BCL-XL inhibitor A-1331852 and STING agonist for 48 hours in culture. (F) Bright field microscopy images of human ENKT lymphoma cell lines treated with A-1331852, ADU-S100, or a combination of both for 48 hours in culture. (G) Subcutaneous xenografting of SNK6 cells into NSG mice and treatment with A-1331852, diABZI, or a combination of both drugs to test the effect of combined BH3 mimicry and STING agonist therapy on human ENKT lymphoma in vivo. (H) Tumor weight of mice treated with A-1331852 or a combination of A-1331852 and diABZI. (I) Representative tumors on the right side of mice treated with A-1331852 or a combination of A-1331852 and diABZI (outlined in yellow).

[0085] Figure 3 This study demonstrates the combination of a BH3 mimic targeting BCL-2 with a STING agonist to enhance apoptosis in cultured human acute myeloid leukemia (AML) cell lines and primary AML patient samples. (A) Western blot of STING in three human AML cell lines. HSP70 blot was used as a loading control. (B) MOLM-13 cells treated with the STING agonist for 24 hours in culture (isogenic non-targeted sgRNA (NT) control and...) TP53 KO variant), MV4;11 (wild type) TP53 ) and THP-1 (mutant) TP53(C) Dose-response curves of cells. (D) Cell viability assay of human AML cell lines treated in culture for 24 hours with a combination of the BCL-2 inhibitor venetoclax and the STING agonist diABZI. (E) Cell viability assay of leukemia blasts (CD117+) from human AML patient samples treated in culture for 48 hours with venetoclax alone (black), diABZI alone (orange), or a combination of venetoclax and diABZI (red). (C) Cell viability assay of normal T cells (CD3+) from human AML patient samples treated in culture for 48 hours with venetoclax alone (black), diABZI alone (orange), or a combination of venetoclax and diABZI (red).

[0086] Figure 4 Analysis of mice xenografted with SNK6ENKT human lymphoma treated with a combination of the STING agonist diABZI and the BCL-XL inhibitor A-1331852 is shown. (A) Subcutaneous tumor volume of mice transplanted with SNK6 human NK T lymphoma cells. (B) Tumor weight of mice treated with the mediator or diABZI at the predetermined ethical endpoint. (C) Weight of metastatic lymph nodes of mice treated with the mediator or diABZI at the ethical endpoint. (D) Cell counts from peripheral blood extracted from mice at the ethical endpoint. (E) Weight of metastatic lymph nodes of mice treated with A-1331852 alone or in combination with diABZI on day 65 post-transplantation. (F) Cell counts from peripheral blood extracted from mice treated with A-1331852 alone or in combination with diABZI on day 65 post-transplantation.

[0087] Figure 5 This study showed that human B lymphoma cell lines lacked STING expression and did not respond to STING agonist treatment in culture. (A) AML and NHL from the Cancer Cell Line Encyclopedia (CCLE), further stratified into NHL subtypes. STING1 Box plots of expression data. (B) Western blot of STING in a mantle of human B lymphoma cell lines. (C) Cell viability assay of human B lymphoma cell lines treated with the STING agonist ADU-S100 for 24 hours in culture. (D) Cell viability assay of human DLBCL cell line DOHH2 treated with S63845 (20 nM) and / or the STING agonist diABZI (100 nM) for 48 hours in culture.

[0088] Figure 6 This study demonstrates that the combination of STING agonists and venetoc induces potent killing of cultured human AML cell lines. Cell viability was measured in syngeneic parental MOLM-13 human AML cells containing non-targeted control sgRNA (NT control) or those mutated using CRISPR / Cas9 to form the TP53-deficient genotype. Cells were treated in culture for 24 hours with a combination of the BCL-2 inhibitor venetoc and the STING agonist ADU-S100 or MSA-2.

[0089] Figure 7 Cell viability assays of MOLM-13 WT (A), OCI-AML3 (B), THP-1 (C), HL-60 (D), KG-1 (E), and TF-1 AML (F) cell lines treated with the BCL-2 inhibitor ABT-199 / venetoclax (thin line, round marker) or compound 4-46 (thick line, square marker) for 48 hours are shown. Data are shown as mean ± SEM.

[0090] Figure 8 and Figure 9 The use of STING agonist compound 4-46 in culture was shown. Figure 8 ) or ABT-199 / Venetok alone ( Figure 9 Cell viability assay of syngeneic human MOLM-13 AML cells treated for 48 hours with non-targeted sgRNA (thick line, square marker), TP53 KO (dashed line, circle marker), STING KO (green line, triangle marker), and Bak / Bax double KO (purple line, diamond marker).

[0091] Figure 10A and 10B This demonstrates the effect of treating the human AML cell line MOLM-13 WT (0-10 µM) in culture for 24 hours with a combination of the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM) and the STING agonist compound 4-46 (0-10 µM). Figure 10A ) and OCI-AML3 ( Figure 10B Activity determination of ).

[0092] Figure 10C and 10D This demonstrates the effect of treating the human AML cell line MOLM-13 WT (0-10 µM) in culture for 24 hours with a combination of the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM) and the STING agonist compound 4-46 (0-10 µM). Figure 10C ) and OCI-AML3 ( Figure 10D The heatmap of the activity determination. Figure 10A The data presented in Figure 10CThe data presented is the same, and Figure 10D The data presented in Figure 10B The data presented is the same.

[0093] Figure 11 ARs show the viability assays of 18 different leukemia blast (LB) cell samples treated with STING agonist compounds 4-46 (thick blue lines; circular markers; 0.0001–10 µM), ABT-199 / venetoclax (dashed red lines; square markers; 0.0001–10 µM), or a combination of two agents at a specified concentration in a 1:1 ratio (thin green lines; triangle markers; 0.0001–10 µM).

[0094] Figure 12A -C shows the results of viability assays of primary human AML cells treated in culture with (A) various concentrations of the STING agonist compound 4-11; (B) the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM); and (C) the combination of the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM) and the STING agonist compound 4-11. Detailed Implementation

[0095] This article provides a combination therapy comprising a BH3 mimic and a STING agonist, and a method for treating a subject with a proliferative disease, such as cancer, wherein the method comprises administering the BH3 mimic and the STING agonist together to the subject.

[0096] While BH3 mimicry therapies have been successfully deployed to treat cancer, their TP53-dependent maximal killing effect allows TP53-mutant cancer cells to survive and proliferate during treatment, leading to relapsed / refractory disease and reduced sensitivity of post-treatment growth to additional rounds of BH3 mimicry therapy. Therefore, a problem in the art is not only the TP53-dependent treatment of cancer cells with BH3 mimicry therapies; a problem in the art is that the population of cancer cells remaining after BH3 mimicry therapy is less susceptible to further treatment and more prone to further and often more severe disease. Against this backdrop, the inventors of the present invention have discovered that, when used as a monotherapy, BH3 mimicry therapies targeting MCL-1 or BCL-2 stabilize and functionally activate TRP53 / TP53, thereby inducing TRP53 / TP53 target genes to maximize apoptotic death in malignant cells, including malignant lymphoma and leukemia cells, thus explaining the TP53 dependence of BH3 mimicry therapy. However, the inventors of this invention have now utilized their discovery that TRP53 / TP53 mutant / deficient cells can still express TRP53 / TP53 target genes (albeit at significantly reduced levels) to find a TRP53-independent mechanism for inducing the expression of genes encoding these BH3-only proteins, a mechanism involving the cyclic GMP-AMP synthase (cGAS) interferon gene stimulating factor (STING) signaling pathway. In particular, the inventors have found that enhanced activation of STING using relevant agonists is highly effective in triggering intrinsic apoptosis in a range of cancer cells in a TP53 / TRP53-independent manner. The inventors have also found that the combination of a BH3 mimic and a STING agonist synergistically enhances the killing of certain cancer cells by utilizing the combination of the promotion of apoptosis in even TP53-deficient cancers through the upregulation of BH3-only proteins in a TP53-independent manner by the STING agonist with a partially TP53-dependent pro-apoptotic protein pathway activated by the BH3 mimic. This new combination not only has an advantage in increasing cancer cell death compared to the administration of either drug alone, but may also open up treatment options in patient groups with cancers treatable by BH3 mimics that were previously unable to receive BH3 mimic monotherapy (e.g., due to DNA damage and / or TP53-mutant cancer cells from previous treatment) and / or who are trying to avoid the development of drug-resistant cancer cell subpopulations that could lead to relapse of malignancy.

[0097] BH3 mimicry drug The combination of BH3 mimics described herein. Several BH3 mimics are known in the art and are considered suitable for use according to this disclosure. Typically, BH3 mimics require binding with very high affinity to large and mostly hydrophobic grooves that support protein-protein interactions between pro-survival BCL-2 proteins (including BCL-2, BCL-XL, BCL-W, MCL-1, A1 / BFL-1, and possibly BCL-B) and the BH3 domains of their pro-apoptotic related molecules. Therefore, all advanced BH3 mimics are characterized by relatively high molecular weight, lipophilicity, and chemical complexity.

[0098] By way of non-limiting examples, suitable BH3 mimicry drugs may include small molecules that target the survival-promoting BCL-2, BCL-XL, and BCL-W, including ABT-737 (CAS No. 852808-04-9), and the closely related drug ABT-263 (navitoclax; CAS No. 923564-51-6), which has the same binding spectrum as ABT-737 but has improved pharmacological properties and is the first BH3 mimicry to enter clinical trials. Other suitable BH3 mimics include the BCL-2 selective inhibitor ABT-199 / venetoclax (marketed as Venclexta® by Genentech USA, Inc. and AbbVie Inc.), which is FDA-approved for the treatment of chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma, or in combination with other drugs for the treatment of newly diagnosed acute myeloid leukemia (AML). The structures of these compounds are shown below: .

[0099] Other BCL-2 inhibitor compounds suitable for the combination therapy described herein include those described in the following publications: WO201114942 A1 and WO 2010065824 A1, which describe alternative heterocyclic compounds of venetoclax; WO2010080478 A1, which describes urea compounds; WO 2010080503 A1, which describes amide compounds; WO2019040573 A1, which describes a series of benzamide compounds; WO 2019185025 A1, which describes trifluoromethyl-substituted sulfonamides; WO 2019210828 A1, which describes nitrobenzene-substituted sulfonamides; and US11053239, which discloses trifluoromethylsulfonates or nitrobenzene-substituted sulfonamides, the disclosures of each of these publications being expressly incorporated herein by reference.

[0100] MCL-1-specific BH3 mimicry compounds suitable for the combination therapies described herein include S63845 (Servier; CAS No. 1799633-27-4) and related compounds S64315 / MIK665 (CAS No. 1799631-75-6), AMG 176 (tapotoclax; Amgen; CAS No. 1883727-34-1), and AZD5991 (AstraZeneca; CAS No. 2143061-81-6). The structures of these compounds are shown below: .

[0101] BH3 mimics, which are high-potency and selective inhibitors of BCL-XL, may also be suitable for the combination therapies described herein, including A-1155463 (AbbVie / Genentech; CAS No. 1235034-55-5), A-1331852 (AbbVie / Genentech; CAS No. 1430844-80-6), and WEHI-539 (Walter and Eliza Hall Institute of Medical Research; CAS No. 2070018-33-4).

[0102] In one embodiment, the BH3 mimic used in the combinations herein is an inhibitor of BCL-2, BCL-XL, BCL-W, or MCL-1, or an inhibitor of any two or more of these pro-survival proteins. In one embodiment, the BH3 mimic used in the combinations herein is a selective inhibitor of BCL-2. In one embodiment, the BH3 mimic used in the combinations herein is a selective inhibitor of MCL-1. In one embodiment, the BH3 mimic used in the combinations herein is a selective inhibitor of BCL-XL. In one embodiment, the BH3 mimic used in the combinations herein is a selective inhibitor of BCL-2, MCL-1, and BCL-XL.

[0103] Other BH3 mimicry compounds are also envisioned, including antibody-drug conjugates comprising a BH3 mimicry drug covalently linked to a monoclonal antibody or a ligand targeting a receptor of interest (as described above), and dendritic polymer-drug conjugates comprising a BH3 mimicry drug encapsulated within or conjugated to the surface of a dendritic polymer (as described above), for use in the combination therapies described herein. Thus, in one embodiment, the BH3 mimicry compound used in the combinations herein comprises one or more BH3 mimicry drugs conjugated to a delivery component. In one embodiment, the BH3 mimicry compound used in the combinations herein comprises an inhibitor of BCL-2, BCL-XL, BCL-W, or MCL-1, or any two or more of these pro-survival proteins, conjugated to a delivery component. In one embodiment, the delivery component is selected from monoclonal antibodies, ligands, and dendritic polymers. In one embodiment, the delivery component is a monoclonal antibody. In one embodiment, the delivery component is a ligand, which in some embodiments is selected from proteins, protein fragments, peptides, small molecules, and nucleic acids. In one embodiment, the delivery component is a dendritic polymer. In one embodiment, the BH3 mimic used in the combinations herein comprises a selective inhibitor of BCL-2 conjugated to the delivery component. In one embodiment, the BH3 mimic used in the combinations herein comprises a selective inhibitor of MCL-1 conjugated to the delivery component. In one embodiment, the BH3 mimic used in the combinations herein is a selective inhibitor of BCL-XL. In one embodiment, the BH3 mimic used in the combinations herein comprises inhibitors of BCL-2, MCL-1, and BCL-XL conjugated to the delivery component. In each of these embodiments, the delivery component may be selected from monoclonal antibodies or ligands and dendritic polymers. For the avoidance of doubt, any reference herein to BH3 mimics shall be understood to cover BH3 mimic compounds.

[0104] It should be understood that BH3 mimicry drugs can be used to treat a wide range of solid cancers, including but not limited to breast cancer, ovarian cancer, neuroblastoma, non-small cell lung cancer, prostate cancer, colorectal cancer, and melanoma. BH3 mimicry drugs can also be used to treat a wide range of so-called "liquid" tumors or blood cancers, including but not limited to certain lymphomas (such as follicular lymphoma and small lymphocytic lymphoma), leukemia subtypes (such as chronic lymphocytic leukemia, acute myeloid leukemia, and acute lymphoblastic leukemia), and multiple myeloma. In one embodiment, the cancer used for treatment with the BH3 mimicry drug combination described herein is TP53 wild-type cancer, TP53 mutant cancer, or cancer comprising both TP53 wild-type and TP53 mutant cell subpopulations. While previous BH3 mimicry therapies have generally been more successful in treating TP53 wild-type cancers, the inventors of this invention have expanded the range of cancers treatable by BH3 mimicry by combining BH3 mimicry therapies with STING agonists that can target cells in a TP53-independent manner. It should be understood that cancers considered treatable by BH3 mimicry, including those specifically mentioned in this paragraph, are candidates for the combination therapies described herein, where those cancer cells also express the STING protein.

[0105] Any suitable pharmaceutically acceptable dose of a BH3 analogue can be used in the combinations described herein. By way of non-limiting example, guidance can be obtained from venetoclax dosing regimens for some indications, in which doses between 10 mg / day and about 400-600 mg / day are appropriate for any suitable period of treatment. These doses can be achieved using any suitable escalation schedule, including but not limited to weekly increases starting at 20 mg / day and increasing to 50 mg / day, 100 mg / day, 200 mg / day, and then to 400 mg / day. In some cases, a faster daily escalation starting at 100 mg / day and increasing to 200 mg / day, 400 mg / day, 600 mg / day may be appropriate. Dosing can be continued on a monthly cycle for any suitable number of cycles, such as 1 to 24 monthly cycles, or 6 to 12 cycles, or 12 to 24 cycles. The initial dosing cycle for cancer treatment using the BH3 mimicry drugs and methods described herein can be calculated by those skilled in the art using any suitable method. In some embodiments, the dose can be calculated based on the relative efficacy and molecular size of a given BH3 mimicry drug compared to venetoclax. In some embodiments, the dose of the BH3 mimicry drug can be equivalent to or substantially equivalent to the dose of venetoclax, or, when used in the combination therapy described herein, can be 5%, 10%, 15%, 20%, 30%, 40%, or 50% lower than the dose of venetoclax (or its equivalent) due to the synergistic effect obtained from the combination of the BH3 mimicry drug and the STING agonist.

[0106] The BH3 analogues used herein can be in any suitable dosage form. In one embodiment, the BH3 analogue is for oral administration. In one embodiment, the BH3 analogue is in the form of tablets for oral administration. In other embodiments, alternative dosage forms, such as capsules, solutions, granules, or powders, can be used. In one embodiment, the BH3 analogue is for intravenous administration.

[0107] STING agonist The combination of STING agonists described herein. Many STING agonists known in the prior art are considered suitable for use according to this disclosure. Typically, STING agonists include small molecule nucleoside analogs, such as synthetic cyclic dinucleotides, as well as emerging second-generation acyclic dinucleotides or non-nucleoside analogs.

[0108] By way of non-limiting examples, suitable STING agonists may include MSA-2 (Merck; CAS No. 129425-81-6), a non-nucleotide small molecule STING agonist; MK-1454 (ulevostinag; Merck; CAS No. 2082743-96-0), a synthetic cyclic dinucleotide STING agonist being investigated for the treatment of solid tumors, lymphomas, and head and neck squamous cell carcinomas; and diABZI compound 3, also known as diABZI. STING agonist 1 (GlaxoSmithKline, CAS No. 2138498-18-5) is a non-cyclic dinucleotide STING agonist; TAK-676 (dazostinag, Takeda, CAS No. 2553413-93-5) is a synthetic cyclic dinucleotide STING agonist; ADU-S100 (also known as MIW815, Novartis, CAS No. 1638750-96-5; now discontinued) is a synthetic cyclic dinucleotide STING agonist studied for its activity against solid tumors and lymphomas; E7766 (CAS No. 2242635-03-4, Eisai, Inc.) Inc.), a macrocyclic-bridged STING agonist studied for activity against solid tumors and lymphomas; MK-2118 (Merck), a small molecule STING agonist studied for activity against solid tumors and lymphomas; and SB 11285 (invoX Pharma), a small molecule STING agonist studied for activity against solid tumors, head and neck squamous cell carcinoma, and melanomas. The structures of some of these drugs are shown below:

[0109] Suitable STING agonists are described in WO 2019 / 219820 (US17,054,850), WO 2021 / 009362 (US17 / 624,137), WO 2021 / 009365 (US17 / 624134), WO 2021 / 119753 (US 17 / 786792), WO 2022 / 266711 and WO 2024 / 130341, the complete disclosure of each of which is incorporated herein by reference. In particular, examples of STING agonist compounds (I) through (V) described in these publications are incorporated herein by reference.

[0110] Specific examples of STING agonists are given in Table 1 below.

[0111] Table 1. Examples of compounds of the present invention

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179]

[0180] In some embodiments, the STING agonist is a compound of formula (I). In the embodiments, the STING agonist is a compound selected from any one of compounds 1-1 to 1-172.

[0181] In some embodiments, the STING agonist is a compound of formula (II). In the embodiments, the STING agonist is a compound selected from any one of compounds 2-1 to 2-25.

[0182] In some embodiments, the STING agonist is a compound of formula (III). In the embodiments, the STING agonist is a compound selected from any one of claims 3-1.1 to 3-24.

[0183] In some embodiments, the STING agonist is a compound of formula (IV). In some embodiments, the STING agonist is a compound selected from any one of compounds 4-1 to 4-112. In some embodiments, the STING agonist is compound 4-11 or 4-46.

[0184] In some embodiments, the STING agonist is a compound of formula (V). In the embodiments, the STING agonist is a compound selected from any one of compounds 5-1 to 5-15.

[0185] In one embodiment, the STING agonist used herein is selected from synthetic cyclic dinucleotides, acyclic dinucleotides, or non-nucleoside small molecules. In one embodiment, the STING agonist used herein is a synthetic cyclic dinucleotide. In one embodiment, the STING agonist used herein is an acyclic dinucleotide. In one embodiment, the STING agonist used herein is a non-nucleoside small molecule.

[0186] Other STING agonist compounds are also envisioned, including antibody-drug conjugates comprising a STING agonist covalently linked to a monoclonal antibody (such as any of the aforementioned STING agonists) and dendritic-drug conjugates comprising a STING agonist encapsulated within or conjugated to the surface of a dendritic polymer (such as any of the aforementioned STING agonists), which could be used in the combination therapies described herein. Therefore, in one embodiment, the STING agonist compound used in the combinations herein comprises one or more STING agonists conjugated to a delivery component. In one embodiment, the STING agonist compound used in the combinations herein comprises a synthetic cyclic dinucleotide, acyclic dinucleotide, or non-nucleoside small molecule conjugated to a delivery component. In one embodiment, the delivery component is selected from monoclonal antibodies, ligands, and dendritic polymers. In one embodiment, the delivery component is a monoclonal antibody. In one embodiment, the delivery component is a ligand, which in some embodiments is selected from proteins, protein fragments, peptides, small molecules, and nucleic acids. In one embodiment, the delivery component is a dendritic polymer. In one embodiment, the STING agonist compound used herein comprises a synthetic cyclic dinucleotide conjugated to a delivery component. In one embodiment, the STING agonist compound used herein comprises a non-cyclic dinucleotide conjugated to the delivery component. In another embodiment, the STING agonist compound used herein comprises a non-nucleoside small molecule conjugated to the delivery component. In each of these embodiments, the delivery component may be selected from monoclonal antibodies, ligands, and dendritic polymers. For the avoidance of doubt, any reference herein to STING agonists shall be understood to encompass STING agonist compounds.

[0187] It should be understood that STING agonists can be used to treat a wide range of solid cancers, including but not limited to prostate cancer, squamous cell carcinoma, non-small cell lung cancer, melanoma, breast cancer, colorectal cancer, fibrosarcoma, and glioma. STING agonists can also be used to treat hematologic malignancies, including but not limited to certain lymphomas and acute myeloid leukemia. In one embodiment, a STING agonist is suitable for treating cancers expressing the STING protein. In one embodiment, a STING agonist is not suitable for treating human B-cell lymphomas because these malignant cells lack STING expression. In one embodiment, a STING agonist is particularly suitable for treating malignant cells expressing STING and carrying a TP53 mutation. In one embodiment, a STING agonist is suitable for treating malignant cells expressing STING in a TP53-independent manner. In one embodiment, the TP53-independence of the STING agonist makes the combinations described herein suitable for induction therapy treatment of a group of patients whose malignant cells have significant DNA damage due to prior radiotherapy and / or chemotherapy and may additionally be intolerant to subsequent high-dose cancer induction regimens. In one embodiment, the TP53-independent nature of the STING agonist makes the combination described herein suitable for patient groups with high levels of TP53-mutant malignant cells, as these cells have severely impaired ability to detect DNA damage and thus initiate apoptosis via conventional innate pathways. It should be understood that cancers considered treatable with STING agonists, including those specifically mentioned in this paragraph, are candidates for the combination therapies described herein, making those cancer cells also readily treatable with BH3 mimicry drugs.

[0188] Any suitable pharmaceutically acceptable dose of the STING agonist can be used in the combinations described herein. STING agonists have been previously studied and used in combination immunotherapies to enhance the host's anti-cancer immune response, and therefore, dosing regimens from such therapies can be used in the combinations and methods of the present invention. By way of non-limiting example, guidance can be obtained from dosing regimens in some indications, in which doses between 10 µg / week and about 6,400 µg / week, or between 10 µg / week and about 1,500 µg / week, or between 10 µg / week and about 3,000 µg / week, or between 90 µg / week and about 3,000 µg / week are suitable for any suitable treatment period. These doses may be suitable for intratumoral administration. In one embodiment, the STING agonist is administered once weekly. In one embodiment, the STING agonist is administered 2-3 times weekly. In one embodiment, the STING agonist is administered every 2-3 weeks. These doses can be administered at any suitable incremental schedule or any suitable on / off schedule, such as a treatment cycle of 3 weeks on / 1 week off per month. The doses can be continued on a monthly cycle for any suitable number of cycles, such as 1 to 6, 1 to 12, or 1 to 24 monthly cycles. Intravenous administration schedules may differ from the weekly doses listed above. The initial dosing cycle for cancer treatment using the STING agonists and methods described herein can be calculated by a person skilled in the art using any suitable method.

[0189] The STING agonist used herein may be in any suitable dosage form. In one embodiment, the STING agonist is in the form of a suspension or solution for intratumoral, intravenous, or subcutaneous administration. In one embodiment, the STING agonist is in a form suitable for intratumoral or intravenous administration. In one embodiment, the STING agonist is in the form of a suspension or solution for intravenous administration. In one embodiment, the STING agonist is for intravenous administration. In one embodiment, the STING agonist is for intratumoral administration.

[0190] combination This document discloses combinations for treating cancer to inhibit its progression, wherein the combinations comprise a BH3 mimic and a STING agonist. In one embodiment, the combination is a synergistic combination. In some embodiments, the combination is in the form of a pharmaceutical composition. The pharmaceutical composition preferably comprises individual formulations of a BH3 mimic and a STING agonist, particularly where the two drugs are suited for different routes of administration and / or according to different dosing schedules. In some embodiments, the combination is in the form of a kit comprising individual formulations of a BH3 mimic and a STING agonist, along with instructions for use of both drugs in combination therapy for a specified indication. In one embodiment, the combination comprising a BH3 mimic and a STING agonist comprises individual formulations of a BH3 mimic (e.g., in an oral dosage form) and a STING agonist (e.g., in an injectable form). In one embodiment, the combination comprises an oral dosage form of a BH3 mimic and a individual formulation of a STING agonist in an intratumoral or intravenous injectable form.

[0191] In some embodiments, the BH3 mimic and the STING agonist are suitable for simultaneous or sequential administration. In one embodiment, the BH3 mimic and the STING agonist are suitable for simultaneous administration. Simultaneous administration may include administration of the BH3 mimic in a dosing regimen that ensures a level of drug activity in the system of a subject being treated when the STING agonist is administered. In some embodiments, the BH3 mimic may be administered daily, and the STING agonist may be administered weekly, with the two dosing regimens envisioned overlapping to ensure that the subject has both drugs in their system simultaneously. Simultaneous administration may include administration of the BH3 mimic and the STING agonist to the subject simultaneously. Thus, in some embodiments, the BH3 mimic may be administered at the same frequency as the STING agonist, such as 1-3 times per week, making the dosing regimens of the BH3 mimic and the STING agonist identical or substantially identical.

[0192] In some embodiments, such as where a delay is needed to reach acceptable plasma levels before oral administration of a BH3 mimic and intravenous or intratumoral administration of a STING agonist, sequential administration of the BH3 mimic and the STING agonist may be appropriate. In other embodiments, it may be sufficient to administer both the BH3 mimic and the STING agonist at different times but within the same therapeutic window, which means that the biological effects of both drugs are not necessarily present simultaneously, but rather that the effects of both drugs on the patient exist within the same therapeutic window.

[0193] It is also envisioned that the combinations described herein may comprise one or more BH3 mimics and one or more STING agonists. In other embodiments, the combinations described herein may further comprise a combination of one or more other chemotherapeutic agents known in the art with a BH3 mimic and a STING agonist.

[0194] In one embodiment, the combination comprises a BH3 mimicry selected from BCL-2 selective inhibitors, BCL-XL selective inhibitors, and MCL-1 selective inhibitors. In another embodiment, the combination comprises a BH3 mimicry selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845. In one embodiment, the combination comprises a STING agonist selected from non-nucleotide small molecules, acyclic dinucleotides, and synthetic cyclic dinucleotides. In yet another embodiment, the combination comprises a STING agonist selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. In one embodiment, the combination comprises a BH3 mimic and a STING agonist, wherein the BH3 mimic is selected from BCL-2 selective inhibitors, BCL-XL selective inhibitors, and MCL-1 selective inhibitors, and the STING agonist is selected from non-nucleotide small molecules, acyclic dinucleotides, and synthetic cyclic dinucleotides. In another embodiment, the combination comprises a BH3 mimic and a STING agonist, wherein the BH3 mimic is selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845; and the STING agonist is selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. In one embodiment, the combination comprises a BH3 mimicry and compounds 4-11, wherein the BH3 mimicry is selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845. In another embodiment, the combination comprises a BH3 mimicry and compounds 4-46, wherein the BH3 mimicry is selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845.

[0195] In one embodiment, the combination comprises a BH3 mimicry drug and a STING agonist, wherein the BH3 mimicry drug comprises a selective MCL-1 inhibitor, such as S63845, and the STING agonist is selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. In another embodiment, the combination comprises a BH3 mimicry drug and a STING agonist, wherein the BH3 mimicry drug comprises a selective BCL-2 inhibitor, such as venetoclax, and the STING agonist is selected from non-nucleotide small molecules, such as MSA-2; acyclic dinucleotides, such as diABZI STING agonist 1; and synthetic cyclic dinucleotides, such as ADU-S100 / MIW815. In one embodiment, the combination comprises a BH3 mimicry with a synthetic cyclic dinucleotide sTING agonist, such as ADU-S100 / MIW815, wherein the BH3 mimicry comprises a selective BCL-XL inhibitor, such as A-1331852. In some embodiments, the cancer treated by these combinations is a hematologic malignancy, optionally selected from acute myeloid leukemia, NKT-cell lymphoma, ENKT lymphoma, and multiple myeloma.

[0196] In one embodiment, the combination is a synergistic combination of a BH3 mimic and a STING agonist, wherein the BH3 mimic is selected from venetoclax, S63845, or A-1331852, and the STING agonist is selected from MSA-2 or ADU-S100. In one embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist MSA-2. In one embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist ADU-S100. In one embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist diABZI. In one embodiment, the combination is a synergistic combination of the BH3 mimic A-1331852 and the STING agonist ADU-S100. In one embodiment, the combination is a synergistic combination of the BH3 mimic S63845 and the STING agonist MSA-2. In one embodiment, the combination is a synergistic combination of BH3 mimic A-1331852 and STING agonist ADU-S100.

[0197] In one embodiment, the combination is a synergistic combination of a BH3 mimic and a STING agonist, wherein the BH3 mimic is selected from venetoclax, S63845, or A-1331852, and the STING agonist is selected from compound 4-46 or compound 4-11. In another embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist, namely compound 4-46 or compound 4-11. In yet another embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist, namely compound 4-46 or compound 4-11. In yet another embodiment, the combination is a synergistic combination of the BH3 mimic venetoclax and the STING agonist, namely compound 4-46 or compound 4-11. In yet another embodiment, the combination is a synergistic combination of the BH3 mimic A-1331852 and the STING agonist, namely compound 4-46 or compound 4-11. In one embodiment, the combination is a synergistic combination of the BH3 mimic drug S63845 and the STING agonist, namely compound 4-46 or compound 4-11. In another embodiment, the combination is a synergistic combination of the BH3 mimic drug A-1331852 and the STING agonist, namely compound 4-46 or compound 4-11.

[0198] Treatment methods and uses This document also discloses methods for treating cancer in a subject or inhibiting cancer progression or tumor growth, the methods comprising administering a BH3 mimic and a STING agonist to the subject. In one embodiment, this document discloses a method for treating cancer in a subject or inhibiting its progression, the method comprising co-administering a BH3 mimic and a STING agonist to the subject. As used herein, the term “co-administering” encompasses both simultaneous and sequential administration of the BH3 mimic and the STING agonist, and is intended to cover any administration schedule that ensures the therapeutic activity of both drugs in the subject at a given time point and / or during a given treatment window. In other words, and to be clear, co-administering means that both drugs must be administered to the subject as part of cancer treatment, but is not intended to administer the two drugs to the patient at exactly the same time and / or in exactly the same dosage form. The methods described herein treat the subject’s cancer by administering a synergistic combination of a BH3 mimic and a STING agonist to the subject.

[0199] As used herein, the term "treatment" means reducing, alleviating, or improving one or more symptoms of a disease or condition, suppressing a disease or condition or its symptoms, alleviating or improving a disease or condition, causing the remission of a disease or condition, and preventing or inhibiting disease progression. In some embodiments, "treatment" includes induction therapy for newly diagnosed malignancies, maintenance therapy for maintaining remission, and / or therapy for treating refractory (unresponsive) or relapsed (progressed after remission) malignancies.

[0200] As used herein, the term "cancer" refers to a disease characterized by growths or tumors resulting from abnormal, uncontrolled cell growth, and includes diseases involving both pre-malignant and malignant cells. Further discussion below focuses on cancers particularly suitable for treatment with the combination therapies described herein.

[0201] As used herein, the term "subject" refers to a human and a non-human mammal. In some embodiments, the subject is a mammal. In other embodiments, the subject is a human. In some embodiments, the subject is a non-human mammal.

[0202] In some embodiments, the methods of treating a subject's cancer or inhibiting its progression described herein comprise administering to the subject a therapeutically effective amount of a BH3 mimic and a therapeutically effective amount of a STING agonist. As used herein, the term "effective amount" in the context of a therapeutically effective amount refers to a dose of an active compound, such as a BH3 mimic or a STING agonist, that prevents disease progression (referred to as "management" in some embodiments), alleviates, significantly reduces, or completely reduces one or more symptoms and / or causes of the treated disease or condition. In some embodiments, the amounts of the BH3 mimic and STING agonist used in the combination therapy described herein are amounts that provide a significant reduction in the clinical symptoms of the treated disease or condition and, in some embodiments, do not cause excessive or intolerable toxic side effects to the subject.

[0203] In one embodiment, the BH3 mimic is administered orally. In one embodiment, the STING agonist is administered intravenously. In one embodiment, the method includes oral or intravenous administration of a combination of the STING agonist and the BH3 mimic. Administration may be simultaneous or sequential as described in the section entitled “Combination” above.

[0204] As further described above, in one embodiment, the BH3 mimic drug is administered at a dose of about 10 mg / day to 600 mg / day, and the STING agonist is administered at a dose of about 50 µg / week to about 6,400 µg / week.

[0205] This article also describes a method for treating a subject with cancer or inhibiting cancer progression or tumor growth, the method comprising: identifying the subject as a candidate for a BH3 mimicry drug therapy; identifying the cancer as cells containing STING protein; and administering both a BH3 mimicry drug and a STING agonist to the subject, wherein the STING agonist and the BH3 mimicry drug are administered sequentially or simultaneously.

[0206] This article further describes a method for treating a subject’s cancer or inhibiting cancer progression or tumor growth, wherein the cancer is refractory or resistant to BH3 mimicry monotherapy, the method comprising: co-administering the BH3 mimicry and the STING agonist to the subject.

[0207] This article further describes a method for inducing an immune response against a tumor or cancer in a subject, the method comprising: co-administering a BH3 mimic drug and a STING agonist to the subject, thereby inducing an antitumor or anticancer immune response in the subject.

[0208] In the methods described herein, the cancer may be a hematologic malignancy or a solid tumor. In some embodiments, the cancer is a TP53 wild-type cancer, a TP53 mutant cancer, or a cancer comprising both a TP53 wild-type cell subset and a TP53 mutant cell subset. In one embodiment, the cancer expresses the STING protein. In some embodiments, the cancer is a TP53 wild-type cancer, a TP53 mutant cancer, or a cancer that is treatable with a BH3 mimicry drug and expresses the STING protein, comprising both a TP53 wild-type cell subset and a TP53 mutant cell subset. In one embodiment, the cancer is a refractory / relapsed cancer. In one embodiment, the cancer is resistant to treatment with BH3 mimicry drug monotherapy. In one embodiment, the cancer is resistant to treatment with BH3 mimicry drug monotherapy and expresses the STING protein.

[0209] When the cancer is a blood cancer, it can be selected from leukemia, malignant lymphoma (ML), multiple myeloma (MM), and myelodysplastic syndrome (MDS), or a relapsed / refractory form of any of these diseases. The cancer can be selected from the following leukemia subtypes: acute myeloid leukemia (AML), including promyelocytic leukemia, chronic myeloid leukemia (CML), and acute lymphoblastic leukemia (ALL), or a relapsed / refractory form of any of these diseases. In some embodiments, the cancer treated by the methods described herein is lymphoma. The lymphoma can be non-Hodgkin's lymphoma (NHL). In such embodiments, non-Hodgkin's lymphoma may be selected from adult T-cell lymphoma, lymphoblastic lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, peripheral T-cell lymphoma, follicular lymphoma, B-cell acute lymphoblastic leukemia, chronic lymphocytic leukemia / small lymphocytic lymphoma, marginal zone lymphoma, Waldenström macroglobulinemia, and mantle cell lymphoma, or any relapsed / refractory form of these diseases. In some embodiments, non-Hodgkin's lymphoma may be selected from adult T-cell lymphoma, lymphoblastic lymphoma, peripheral T-cell lymphoma, follicular lymphoma, B-cell acute lymphoblastic leukemia, chronic lymphocytic leukemia / small lymphocytic lymphoma, marginal zone lymphoma, Waldenström macroglobulinemia, and mantle cell lymphoma, or any relapsed / refractory form of these diseases. In some embodiments, non-Hodgkin's lymphoma does not include B-cell lymphoma and Burkitt's lymphoma. In some embodiments, the cancer is selected from the following blood cancers: acute myeloid leukemia (AML), including TP53-mutant AML, natural killer / T-cell lymphoma (NKTL), extranodal NK / T-cell lymphoma (ENKTL), TP53-mutant NKTL, chronic lymphocytic leukemia (CLL), and multiple myeloma. In some embodiments, the cancer is selected from the following blood cancers: acute myeloid leukemia (AML), including TP53-mutant AML, extranodal natural killer / T-cell lymphoma (ENKTL), and TP53-mutant NKTL.

[0210] In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor is selected from small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, melanoma, breast cancer, ovarian cancer, neuroblastoma, prostate cancer, and colorectal cancer. Solid tumors are not particularly limited, as long as they express the STING protein and are susceptible to BH3 mimic-induced apoptosis.

[0211] In some embodiments, the cancer is not breast cancer.

[0212] This article also discloses the use of a BH3 mimic and a STING agonist in the preparation of a medicament for treating cancer in a subject, wherein the BH3 mimic and the STING agonist are administered co-administered. As discussed above, the co-administration may be sequential or simultaneous.

[0213] The use of a STING agonist in the preparation of a medicament for use in combination with a BH3 mimicry to treat cancer in a subject is also described. The combination administration may be sequential or simultaneous. The use of a BH3 mimicry in the preparation of a medicament for use in combination with a STING agonist to treat cancer in a subject is also described.

[0214] As used herein, unless the context otherwise requires, the term “dosage” refers to the mass of an active substance administered to a subject in milligrams (mg) or micrograms (µg) per kilogram (kg) of the subject’s body weight.

[0215] In some embodiments, the term "about" refers to an amount within ±10%, ±5%, or ±2% of the value modified by the term.

[0216] It should be understood that any numerical range described herein is intended to include all subranges included therein. For example, a range “x to y” or “between x and y” is intended to cover all subranges between x and y, and also includes the range endpoints x and y.

[0217] As used herein, unless otherwise specifically stated, the singular forms “a / an” and “the” may refer to the plural article.

[0218] Salts of the compounds described herein are preferably pharmaceutically acceptable, but it will be understood that non-pharmaceutically acceptable salts also fall within the scope of this disclosure, for example, because these salts can be used as intermediates in methods for preparing pharmaceutically acceptable salts or methods that do not require administration to a subject.

[0219] The term “pharmaceutical acceptable” can be used to describe any salt, solvate, tautomer, N-oxide, stereoisomer and / or prodrug, or any other compound that, when administered to a subject, provides (directly or indirectly) the compound, or its active metabolites or residues, and is typically harmless to the subject.

[0220] Suitable pharmaceutically acceptable salts include, but are not limited to, pharmaceutically acceptable inorganic acids such as salts of hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfonic acid, and hydrobromic acid; or pharmaceutically acceptable organic acids such as salts of acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, malic acid, citric acid, lactic acid, mucoic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edemaic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid.

[0221] Alkali salts include, but are not limited to, salts formed from pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, alkylammonium (e.g., salts formed from triethylamine), salts formed from alkoxyammonium (e.g., salts formed from ethanolamine), and salts formed from ethylenediamine, choline, or amino acids (e.g., arginine, lysine, or histidine). General information regarding the types of pharmaceutically acceptable salts and their formation is known to those skilled in the art and is described in general texts such as “Handbook of Pharmaceutical Salts”, PH Stahl, CG Wermuth, 1st edition, 2002, Wiley-VCH.

[0222] When the compounds are solids, those skilled in the art will understand that the compounds, agents, and salts of the present invention can exist in different crystalline or polymorphic forms, all of which are within the scope of the present invention and the specified chemical formulas.

[0223] This invention includes all crystalline forms of the compounds described herein, including anhydrous crystalline forms, hydrates, solvates, and mixed solvates. All polymorphs are within the scope of this invention if any of these crystalline forms exhibit polymorphism.

[0224] The compounds described herein are intended to cover (where applicable) both solvated and non-solventized forms. Therefore, the compounds described herein include those with indicative structures, including hydrated or solvated forms, as well as non-hydrated and non-solventized forms.

[0225] The compounds described herein, or their salts, tautomers, N-oxides, polymorphs, or prodrugs, may be provided in the form of solvates. The solvates contain stoichiometric or non-stoichiometric amounts of solvent and may be formed during crystallization using pharmaceutically acceptable solvents such as water, alcohols (e.g., methanol, ethanol, or isopropanol), DMSO, acetonitrile, dimethylformamide (DMF), acetic acid, etc., by non-covalent bonding or by occupying pores in the lattice. When the solvent is water, a hydrate is formed; when the solvent is an alcohol, an alcohol is formed. Solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. Generally, for the purposes of this invention, the solvated form is considered equivalent to the non-solvated form.

[0226] Basic nitrogen-containing groups can be quaternized with the following reagents: C1-6 alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates, such as dimethyl sulfate and diethyl sulfate; and others.

[0227] Nitrogen-containing groups can also be oxidized to form N-oxides.

[0228] The compounds described herein, or their salts, tautomers, N-oxides, solvates, and / or prodrugs, which form crystalline solids, may exhibit polymorphism. All polymorphic forms of the compounds, salts, tautomers, N-oxides, solvates, and / or prodrugs are within the scope of this invention.

[0229] The compounds described herein can exhibit tautomerism. Tautomers are two interchangeable forms of a molecule that are normally present in equilibrium. Any tautomer of the compounds described herein should be understood to be within the scope of this invention.

[0230] The compounds described herein may contain one or more stereocenters. All stereoisomers of the compound of formula (I) are within the scope of this invention. Stereoisomers include enantiomers, diastereomers, geometric isomers (E- and Z-alkene forms and cis- and trans-substituted modes), and trans-restricted isomers. In some embodiments, the compound is a stereoisomerically enriched form of the compound of formula (I) at any stereocenter. The enrichment of the compound in one stereoisomer may be at least about 60%, 70%, 80%, 90%, 95%, 98%, or 99% of the enrichment in another stereoisomer.

[0231] The compounds described herein, or their salts, tautomers, solvates, N-oxides, and / or stereoisomers, can be isotopically enriched with respect to one or more isotopes of the atoms present in the compound. For example, the compound can be enriched using one or more of the following trace isotopes: 2H, 3H, 13C, 14C, 15N, and / or 17O, preferably 2H. An isotope can be considered enriched when its abundance is greater than its natural abundance.

[0232] A "prodrug" is a compound that may not fully meet the structural requirements of the compounds provided herein, but is modified in vivo to produce a compound of formula (I) provided herein after administration to a subject or patient. For example, a prodrug can be an acylated derivative of a compound as provided herein. Prodrugs include compounds in which a hydroxyl, carboxyl, amine, or thiol group is bonded to any group and is cleaved upon administration to a mammalian subject to form a free hydroxyl, carboxyl, amino, or thiol group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, phosphate, and benzoate derivatives of alcohol and amine functional groups in the compounds provided herein. Prodrugs of the compounds provided herein can be prepared by modifying the functional groups present in the compound in such a way that the modifier is cleaved in vivo to produce the parent compound.

[0233] Prodrugs comprise compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues is covalently linked to the free amino and amide groups of a compound of formula (I). The amino acid residues include 20 naturally occurring amino acids, typically represented by three-letter symbols, and also include 4-hydroxyproline, hydroxylysine, desmodium, isodesmodium, 3-methylhistidine, valine, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also comprise compounds in which carbonates, carbamates, amides, and alkyl esters are covalently bonded to the above substituents of the compound described herein via a carbonyl carbon prodrug side chain.

[0234] Example Materials and Methods Cancer cell lines and tissue cultures mice All experiments involving animals were conducted in accordance with the Melbourne Directorate Animal Ethics Committee guidelines, as well as the ethics committee of the Walter Elizabeth Institute for Medical Research. Regarding the use of... Eµ-Myc In vivo experiments with lymphoma cells, on day 0... C57BL / 6 Cas9 KI / KI or Rag1 - / -Mice were administered 1 x 10 intravenous (IV) injections. 6 AH15A wild type or Trp53 KO cells. Mice were treated daily via intravenous injection of 25 mg / kg S63845 (Chemgood, C1370) or a mediator (50 mM PBS containing 2% vitamin E (Sigma, 57668)) from day 4 to day 8 post-transplantation. On days 4, 7, and 11 post-transplantation, mice were treated via intravenous injection of 1.5 mg / kg diABZI Sting agonist compound 3 (diABZI; SYNthesis Med Chem) or a mediator (saline containing 40% polyethylene glycol (PEG)-400 (Sigma, P3265)). Lymphoma development in mice was monitored by experienced animal technicians who were unaware of the nature of the transplanted tumor cells and the treatments applied at predetermined ethical endpoints. Retroorbital blood samples were analyzed by Advia to obtain blood cell counts and to weigh enlarged organs (spleen, lymph nodes, thymus). For in vivo xenograft experiments, NOD-SCID-g was injected on day 0. c – / – (NSG) Subcutaneous injection 2 x 10 6 SNK6 human NKT lymphoma cells were transplanted. Mice were treated orally with 50 mg / kg of A-1331852 (a friendly gift from Professor G. Lessene of the Walter Elizabeth Institute for Medical Research (WEHI)) or a mediator (2.5% DMSO, 10% EtOH, 27.5% PEG-400, 60% Phosal 50 PG (Lipoid)) on days 7, 10, and 13 post-transplantation. Mice were treated intravenously with 1.5 mg / kg of diABZI or a mediator (0.9% saline containing 40% PEG-400). Weight was measured every 4–5 days by an experienced animal technician who was unaware of the applied treatment and used an equation. π / 6 x length x width 2 To calculate tumor volume. When the tumor reaches 0.5 cm... 3Mice were euthanized. Due to the changing tumor location in mice treated with A-1331852 or a combination of A-1331852 and diABZI, the volume of the tumors could not be accurately measured using calipers; these tumors appeared later than in mice treated with either the mediator alone or diABZI. Therefore, these mice were collected on day 65 post-transplantation, and tumor weights were directly compared. For all mice, retroorbital blood samples were analyzed by Advia to obtain blood cell counts, and the spleen, enlarged lymph nodes, and tumors were weighed.

[0235] Cancer cell lines and tissue cultures From the origin Eµ-Myc Mice derived from tumors in transgenic mice (C57BL / 6) Eµ-Myc Lymphoma cell lines AH15A, AF47A, and 560 were cultured in FMA medium, which consisted of high-glucose DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich F9423), 100 µM L-asparagine (Sigma-Aldrich A4284), 50 µM β-mercaptoethanol (Sigma-Aldrich M3148), 100 U / mL penicillin, and 100 mg / mL streptomycin (Gibco 15140122). Mouse cells were maintained at 37°C in 10% CO2. Cells were derived from... Eµ-Myc / dCas9a-SAM + / - / sgBcl-2Mouse double-hit lymphoma cell lines 214DHL, 216DHL, and 270DHL (Deng, Diepstraten et al. 2022) were derived from mouse tumors and cultured as described above. Human Burkitt lymphoma cell line BL2 was provided by the University of Birmingham, UK, and cultured in RPMI-1640 medium supplemented with 10% FBS, 1 mM sodium pyruvate (Gibco, catalog number 11360070), 2 mM L-glutamine (Gibco, catalog number 25030081), 50 µM α-thioglycerol (Sigma-Aldrich, catalog number M-6145), 100 U / mL penicillin, and 100 µg / mL streptomycin. Human DLBCL cell line DOHH2 and AML cell lines MV4;11, THP-1, and MOLM-13 were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS, 100 U / mL penicillin, and 100 µg / mL streptomycin. Human NKT lymphoma cell lines SNK6, MEC04, and SNT15 were provided by the University of Birmingham, UK, and were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated human serum (Sigma-Aldrich, catalog number H4552), 1 mM sodium pyruvate, 2 mM L-glutamine, 700 U / mL human IL-2 (PeproTech, catalog number 200-02), 100 U / mL penicillin, and 100 µg / mL streptomycin. Human lymphoma and leukemia cell lines were maintained at 37°C with 5% CO2 and validated by STR profiling at the Australian Genomics Research Facility (AGRF). HEK293T cells were cultured in DMEM supplemented with 10% FBS, 100 U / mL penicillin, and 100 mg / mL streptomycin at 37°C with 10% CO2. All cell lines were passaged for <3 months and periodically tested for mycoplasma negativity (Lonza MycoAlert).

[0236] Human AML cell lines MOLM-13 WT, MOLM-13, OCI-AML3, THP-1, HL-60, KG-1, and TF-1 were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS, 100 U / mL penicillin, and 100 µg / mL streptomycin. Human leukemia cell lines were maintained at 37°C with 5% CO2 and validated by STR profiling at the Australian Genome Research Facility (AGRF). All cell lines were passaged for <3 months and periodically tested for mycoplasma negativity (MycoAlert, Lonza).

[0237] The generation of CRISPR / Cas9 gene knockout lymphoma and leukemia cell lines CRISPR / Cas9 gene editing was performed using lentiviral constructs of FuCas9-Cherry (Addgene plasmid number 70182) and FgH1tUTG (Addgene plasmid number 70183) as previously described (Diepstraten, Young et al. 2023). For all experiments involving cell lines producing CRISPR / Cas9, expression was used. Cas9 and targeting people BIM (For mouse cell lines) or mice Bim Parental cells containing the non-targeted (NT) control sgRNA (for human cell lines) were used as control (NT control) cells. Successful knockout of the targeted protein was confirmed by Western blotting.

[0238] Human AML knockout cell lines were generated as previously described (Thijssen, Diepstraten, Moujalled et al., *Blood*, 2021). In short, sgRNA guides and non-targeted control guides targeting human TP53, STING, BAX, and BAK were obtained or synthesized (using integrated DNA technology) from the Sanger WholeGenome CRISPR Arrayed Library and cloned into pKLV-U6gRNA(Bbsl)-PGKpuro2ABFP (Addgene plasmid number 50946). All lentiviruses were generated in 293T cells (ATCC number CRL-3216), and the cell lines were transduced using established protocols. Stable Cas9-expressing human target cells were generated by transduction with FuCas9Cherry (Addgene plasmid number 70182). Subsequently, the human cell lines were transduced with lentiviral supernatants containing the corresponding target sgRNAs. Successful knockout was confirmed using Western blot analysis and / or targeted next-generation sequencing.

[0239] Cell line death assay Cells were spaced at 3 x 10 4 Two copies of each cell / well were plated into 96-well flat-bottom plates, and drugs were added at the specified concentrations. The following drugs were used: S63845, A-1331852, ABT-199 / venetoclax (ActiveBiochem, No. A-1231), ADU-S100 (MedChemExpress, No. HY-12885A), MSA-2 (MedChemExpress, No. HY-136927), TPCA-1 (MedChemExpress, No. HY-10074), GSK8612 (MedChemExpress, dissolved in DMSO), diABZI Sting agonist compound 3, doxorubicin (Ebewe Interpharma), and ionomycin (Sigma-Aldrich, No. I9657). At the designated time points, cells were transferred to round-bottom 96-well plates, spun down at 1,500 rpm for 5 min, and resuspended in annexin V binding buffer containing annexin V-A647 (in-house manufactured, 1:2000) and propidium iodide (PI; 1 µg / mL). Live cell counts (annexin V / PI double negative) were performed using an LSR II flow cytometer (BD Biosciences). Data analysis was performed using FlowJo v10 and GraphPad Prism v9.

[0240] The combination of compounds 4-46 / ABT-199. Cells were loaded at 2 x 10⁻⁶. 5 Cells were plated in triplicate per well into 96-well U-shaped plates, and drugs were added at the specified concentration. The drugs used were compound 4-46 and ABT-199 / venetoclax (Chemgood). After 48 hours, cells were harvested and spun at 300 g for 5 minutes, then resuspended in DAPI or PI at a dilution of 1:2000 for cell viability quantification using Cytoflex (Beckman Coulter). Data analysis was performed using FlowJo v10 and GraphPad Prism v9.

[0241] Protein blot Western blotting was performed as previously described (Diepstraten, Young et al. 2023). Cells were pretreated with 25 µM of the broad-spectrum caspase inhibitor QVD-O-Ph for 15 min (MedChemExpress, HY-12305) and then treated with S63845, STING agonist, or Nutlin-3a (MedChemExpress, HY-10029). Protein concentrations were measured using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, 23225). 10 µg of protein was loaded onto each sample. The antibodies used are listed in Table 2 below. Table 2 - List of antibodies used for Western blotting

[0242]

[0243] Quantitative reverse transcriptase PCR (qRT-PCR) RNA extraction, cDNA synthesis, and TaqMan qRT-PCR were performed as previously described (Diepstraten, Young et al. 2023). Cells were pretreated with 25 mM QVD-O-Ph caspase inhibitor for 15 min, followed by treatment with S63845, STING agonist, or Nutlin-3a. Data were analyzed using the ΔΔCt method, and relative to the housekeeping control gene count for each sample (…). Hmbs Normalization. Data was plotted in GraphPad Prism v9. Table 3 below lists the TaqMan probes: Table 3 - List of TaqMan probes

[0244]

[0245] Cell cycle and proliferation analysis To determine cell number and viability for cell proliferation analysis, an equal volume of 0.4% trypan blue (BioRad, Serial No. 1450021) was added to the sample, and cells were counted on a TC20 automated cell counter (BioRad). Data were plotted in GraphPad Prism v9. For cell cycle analysis, cells were treated with DMSO or diABZi for 24 hours, then collected and fixed on ice for 30 minutes using eBioscience Foxp3 / transcription factor staining buffer (Thermo Fisher Scientific, Serial No. 00-5523-00). Cells were then resuspended in 1x permeabilization buffer and incubated at 4°C for at least 48 hours. Cells were then spin-down and resuspended in PBS containing 5% FBS and 0.5 µg / mL DAPI (Sigma, Serial No. D9542) and analyzed on an LSR II W flow cytometer (BD Biosciences). Data were analyzed using FlowJo v10.

[0246] Imaging Bright-field imaging of NKT lymphoma cells was performed using a ZOE fluorescence cell imaging system (Bio-Rad Laboratories).

[0247] AML patient samples Bone marrow and peripheral blood samples were collected from patients with newly diagnosed or morphologically relapsed AML (16 samples) or myelodysplastic syndromes with excessive blasts (2 samples) treated at two hospitals in Australia (The Peter MacCallum Cancer Centre and The Alfred Hospital). Mononuclear cells were derived using a Ficoll density gradient. Freshly processed or thawed cryopreserved cells were used in drug sensitivity assays. Cells were cultured in Stempsan SFEM medium (StemCell Technologies, No. 09650) supplemented with Stemreginin (500 nM; StemCell Technologies, No. 72354), UM171 (35 nM; Selleckchem, No. S7608), and recombinant human cytokines IL-3 (1 ng / mL; R&D Systems, No. 203-IL), IL-6 (2 ng / mL; R&D Systems, No. 206-IL), FLT3-ligand (5 ng / mL; R&D Systems, No. 308-KFN), and stem cell factor (5 ng / mL; R&D Systems, No. 255-SC). 15,000 to 50,000 cells were seeded into 96-well U-shaped plates and cultured at 37°C and 5% CO2 for 48 hours at the specified drug concentrations. Cell viability was assessed by flow cytometry using a NovoCyte Quanteon (Agilent Technologies). First, cells were stained with antibodies detecting extracellular markers to enable lineage identification (CD45-FITC, CD34-PerCP-Cy5, CD117-PE-Cy7, HLA-DR-APC-Cy7, CD33-BV650, CD14-Alexa Fluor 700, CD64-BV785, CD3-BV421, CD19-PE), followed by PI staining to assess viability.

[0248] Results - During cultivation Combination therapy of BH3 mimics and STING agonists enhances the killing effect on TRP53-deficient lymphoma cells. To determine the potential of STING agonists to enhance the activity of BH3 mimicry drugs, particularly for TP53-deficient hematologic malignancies that frequently relapse after BH3 mimicry drug monotherapy, combinations of the BH3 mimicry drug S63845 (i.e., an MCL-1 specific inhibitor) with STING agonists (ADU-S100, MSA-2, or diABZI) were used in culture against parental (NT control) and [other organisms]. Trp53 KO Eµ-Myc The lymphoma cells were treated.

[0249] Figure 1 A shows the results of treating culture with a combination of the MCL-1 inhibitor S63845 (AH15A: 100 nM, AF47A: 200 nM) and the STING agonist ADU-S100 (AH15A: 2 µg / mL; AF47A: 10 µg / mL), MSA-2 (AH15A: 12 µM, AF47A: 34 µM), or diABZI (AH15A: 20 nM, AF47A: 1 µM) for 24 hours as a syngeneic non-targeted sgRNA (NT) control. Trp53 KO and Bak / Bax Double KO mice Eµ-Myc Cell viability assay for lymphoma cell lines.

[0250] Figure 1 B shows the syngeneic NT control lymphoma cells relative to their... Trp53 Assay for KO derivatives in cell competition in culture. Lymphoma cells were mixed at a 50:50 ratio and treated for 7 days with DMSO (grey), S63845 (red; 50 nM), ADU-S100 (blue; 1 µg / mL), or a combination of S63845 and ADU-S100 (purple), and the proportion of lymphoma cells of each genotype was monitored by flow cytometry over time. The results at each time point were plotted. Trp53 The proportion of KO lymphoma cells.

[0251] Figure 1 C shows the cell viability assay of mouse DHL cell lines treated for 24 hours in culture with the BCL-2 inhibitor venetoc (10 nM) in combination with the STING agonists ADU-S100 (DHL214: 2 µg / ml; DHL270: 5 µg / ml), MSA-2 (DHL214: 17 µM, DHL270: 34 µM), or diABZI (DHL214: 50 nM, DHL270: 100 nM).

[0252] Figure 1 D and 1E show transplanted syngeneic NT control (D) or Trp53 KO(E)AH15A mice Eµ-Myc Lymphoma cells were then treated with a mordant (black), the STING agonist diABZI (blue), the MCL-1 inhibitor S63845 (yellow), or diABZI with S63845 (pink). Rag1Survival curves of mice. Boxes indicate when diABZI (blue) and S63845 (yellow) were administered.

[0253] Figure 1 A shows that the combination therapy enhances the efficacy in parental lymphoma cells and [the following is a separate, unrelated statement:] in an additive and synergistic manner. Trp53 KO Eµ- Myc Killing of lymphoma cells in the culture of both. Bak / Bax Double-KO lymphoma cells were fully resistant, confirming that the combined activity was apoptosis-dependent. Notably, this drug combination inhibited cell competition in culture compared to treatment with BH3 mimicry drugs alone. Trp53 KO's outward growth exceeds the inhibition of parental lymphoma cells. Figure 1 B. In BCL-2-dependent mouse DHL cells, additive killing can be achieved in culture by combining a STING agonist with a BCL-2 selective inhibitor, venetoclax. Figure 1 D).

[0254] Next, the safety and efficacy of the combined BH3 mimicry and STING agonist therapy were examined in vivo. To depict the direct tumor cell-intrinsic effects of the STING agonist on cancer cells in vivo, parental (NT control) or... Trp53 KO E µ-Myc Lymphoma cells transplanted into cells lacking mature T cells and B cells Rag1 In defective mice, mice were treated with a BH3 mimic targeting MCL-1, the STING agonist diABZI, or a combination of both. Mice receiving parental lymphoma cells and treated with diABZI had significantly longer survival times than mice treated with the mediator. Figure 1 E; median survival was 16 days versus 22 days. Furthermore, 3 out of 5 mice (60%) in the MCL-1 inhibitor + STING agonist combination group survived 90 days post-transplantation, compared to only 1 out of 6 mice (17%) receiving MCL-1 inhibitor monotherapy. For mice transplanted with highly invasive... Trp53 Mice with KO lymphoma cells ( Figure 1 F), compared to MCL-1 inhibitors alone, the combination of an MCL-1 inhibitor and a STING agonist significantly increased survival (median survival: 26 days vs. 20 days). Overall, the drug combination was well-tolerated with no adverse events. These results suggest that adding a STING agonist to BH3 mimicry therapy can enhance the efficacy against parental (wild-type) malignant cells by directly targeting them. Trp53 )and Trp53It kills mutant cancer cells, and this prolongs their survival without relying on the function of immune effector cells.

[0255] BH3 mimicry and STING agonists directly enhance apoptosis in human NK / T lymphoma cells. Intrinsic expression of the STING protein is essential for the STING agonist to function and become activated. Therefore, to determine which human hematologic malignancies this approach should ideally target, studies were conducted in human cancer cell lines from the Encyclopedia of Cancer Cell Lines (CCLE). STING mRNA expression ( Figure 5 A). Lymphoma cell lines often have STING The expression showed a bifurcation distribution, with B-cell lymphomas exhibiting low expression and T-cell lymphomas showing high expression. Western blot analysis of STING protein expression in a mantle of human B-cell lymphoma (diffuse large B-cell lymphoma and Burkitt lymphoma) cell lines confirmed that these cells did not have detectable STING expression. Figure 5 Therefore, none of these lymphoma cell lines showed any sensitivity to the STING agonist.

[0256] Because ENKT lymphoma is highly aggressive and the prognosis for patients is currently very poor, the inventors explored the potential of STING agonists to treat this malignancy. STING agonists activate the cGAS / STING pathway in ENKT lymphoma cell lines during culture. Figure 2 B), and showed potent antiproliferative activity when used as a monotherapy in culture. Figure 2 C). The use of the defective TP53 pathway in culture of three ENKT lymphoma cell lines (including those with proven...) TP53 Highly efficient killing of a mutant lymphoma cell line (MEC04) can be achieved in culture by using a combination of treatment with the STING agonist and the BCL-XL-targeting BH3 mimic A-1331852. Figure 2 DF). To test the effects of combined BH3 mimicry and STING agonist therapy on human ENKT lymphoma in vivo, SNK6 cells were subcutaneously xenografted into NSG mice, and the mice were treated with A-1331852, diABZI, or a combination of the two drugs. Figure 2 G). Tumors were first detected in mice treated with either the mediator alone or diABZI at 32 ± 2 days (mean ± SD) post-transplantation (for mediator-treated mice) and 35 ± 1 days (for diABZI-treated mice).

[0257] These mice were analyzed when tumor volume reached a predetermined ethical endpoint, with two groups ( Figure 4), tumor weight at the endpoint ( Figure 4 B) Weight of lymph node metastases ( Figure 4 C) or blood count (except for a slight decrease in platelet count in mice treated with the causative agent) Figure 4 D) There was no significant difference between the two groups. In mice treated with A-1331852, tumors were first detected on average 35 ± 2 days post-transplantation, while tumors in mice treated with the combination of A-1331852 and diABZI remained undetectable on average for 40 ± 2 days, with 2 / 6 of the mice remaining tumor-free until 55 days post-transplantation. The tumors in both groups eventually grew outward in shape and location, making standard external estimates of tumor volume (described in “Methods”) no longer considered an accurate measure. Therefore, all mice treated with A-1331852 alone or in combination with diABZI were collected on the same day (65 days post-transplantation), at which point the first mouse in each group was judged to have reached the ethical endpoint in tumor size, allowing for direct comparison of tumor weight. The inventors found that mice treated with a combination of A-1331852 and diABZI had significantly reduced tumor weight compared with mice treated with A-1331852 alone (1.3 ± 0.3 g (mean ± SEM) vs. 3 ± 0.4 g; Student's test, p = 0.0025). Figure 2 H, I). Additionally, 6 / 6 of the mice treated with A-1331852 had enlarged metastatic axillary lymph nodes on the same side as the tumor xenograft, while 3 / 6 of the mice treated with combination therapy did not have lymph node metastasis, and the remaining 3 / 6 showed only mildly enlarged axillary lymph nodes. Figure 4 E). Blood counts were similar across these treatment groups ( Figure 4 F). Finally, tumors were observed to extend significantly into the peritoneum in 2 / 6 of mice treated with A-1331852 alone, resulting in the formation of secondary masses within the peritoneal cavity. This phenomenon was not observed in any mice treated with a combination of A-1331852 and diABZI. Overall, these results indicate that the combination of a BH3 mimic and a STING agonist can significantly inhibit the outward growth of aggressive human ENKT lymphoma cells, independent of the function of immune effector cells in vivo.

[0258] BH3 mimicry drugs and STING agonists synergistically enhance killing effects on human AML cell lines and AML patient samples. According to data from CCLE, myeloid cancer cell lines show the highest levels of activity among blood cancers. STING Cell lines expressing STING protein. Robust expression of STING protein was confirmed in AML cell lines. Figure 1A), and all cell lines were used in culture as single agents of the STING agonist ADU-S100, MSA-2, or diABZI for highly effective killing (A). Figure 1 B), including TP53 / KRAS mutant THP-1 cells. Combining STING agonists with the BCL-2 inhibitor venetoclax to enhance… TP53 The activity of mutant AML cells has a strong theoretical basis, as these cells are known to have suboptimal responses to BH3 mimics alone or even in combination with chemotherapy. The combined activation of STING and inhibition of BCL-2 have both been shown to be effective in culture, regardless of... TP53 How is the mutation state? Figure 3 C). Next, for those with different genomic profiles, including TP53 defect( Figure 3 Samples from AML patients (D) were examined. Interestingly, 4 / 7 of the samples were highly sensitive to diABZI alone in culture, while the remaining 3 / 7 showed highly effective killing upon treatment with a combination of diABZI and venetoclax in culture. Figure 3 D). Notably, STING agonists, either alone or in combination with venetoclax, were able to kill AML patient samples in culture that showed the lowest sensitivity to treatment with venetoclax alone. Figure 3 D). Interestingly, normal T cells isolated from AML patient samples were resistant to killing by diABZI alone in culture, but the combination of venetoclax and diABZI could efficiently kill these cells in culture. Figure 3 E). These results indicate that STING agonists, and their combination with venetoclax, are beneficial for patients with AML, including those at high risk. TP53 Mutations can play an important clinical role.

[0259] These data were analyzed for synergistic effects using a simple Bliss scoring system. This scoring system assumes that, for a given drug, namely a BH3 mimic (A) and a STING agonist (b), each drug produces its anticancer effect by targeting different pathways, and that these pathways are not mechanistically connected except for the response outcome. The percentage produced, often referred to as the inhibition rate, is typically measured as the percentage of cancer cells that die after drug treatment. The Bliss independence principle was considered to determine the combined effect of the two drugs. Therefore, the Bliss-predicted inhibition rate... y ab It can be calculated using Equation 1.

[0260] Equation 1 y ab = ya + y b – y a y b ,in y a and y b The dosage is a Individual drug A and dosage are b The inhibition rate observed with drug B alone. If the inhibition rate is observed at the combined doses of drug A and drug B... y ab Greater than the inhibition rate predicted by Bliss y ab At the specific dose combination, the drug combination effect is considered synergistic. That is, if the result <1, the combination is synergistic; if =0, it is independent; and if >1, the combination is antagonistic. Table 4 lists the results of various Bliss scores. The results show that a synergistic effect has been observed in all results tested so far. Some combinations showed scores superior to those of other drugs, and the effect depended on the potency of the tested drug and / or the sensitivity of the cell line. The results of the Bliss scores of the control parental cells (NT) are compared with... Figure 1-6 The percentage of live cells was compared, showing that the killing effect was independent of cell type.

[0261] Table 4 - Bliss Rating

[0262]

[0263] discuss The inventors of this invention have demonstrated that doses of BH3 mimics that induce MOMP in human and mouse lymphoma and leukemia cell lines induce TP53 / TRP53 itself. This stabilized TP53 / TRP53 is functional, potently inducing the expression of genes for pro-apoptotic BH3 proteins regulated by TP53, and enhancing the death of malignant cells that might otherwise fail to reach the apoptosis threshold. Importantly, this phenomenon occurs with both MCL-1 inhibitors and BCL-2 inhibitors, thus demonstrating a class effect of BH3 mimics.

[0264] Having understood the conditions required for efficient apoptosis induction in TP53-mutant hematological malignancies, the inventors identified that activation of the cGAS / STING pathway can enhance the expression of pro-apoptotic BH3 protein in a TP53-independent manner, thereby amplifying pro-apoptotic signaling in cells co-treated with BH3 mimicry drugs. Combinations of STING agonists with immunotherapies aimed at enhancing the host's anti-cancer immune response have entered clinical trials; however, to date, these studies have reported limited efficacy from stimulation of the host immune response. Importantly, however, these drugs have demonstrated tolerability in humans.

[0265] In the examples above, this class of STING agonist drugs was repurposed, demonstrating that both synthetic cyclic dinucleotide drugs and small-molecule activators of STING can kill mouse and human hematological malignancies by inducing intrinsic apoptosis in tumor cells in vitro and in vivo, without activating the immune system. The degree of killing of lymphoma and leukemia cells is not limited by TP53 type, but the presence of STING protein is required. Rigorous characterization of this pathway reveals that TBK1 and IRF3 are essential for efficient killing of lymphoma and leukemia cells, while the triggering of NFκB signaling is unnecessary, but importantly, it is not antagonistic to cell killing. Understanding the key regulators essential for apoptosis induction in the cGAS / STING pathway may help patients choose therapies and improve pro-apoptotic STING signaling.

[0266] Importantly, in this study, the inventors demonstrated that combining a STING agonist with a BH3 mimic targeting MCL-1, BCL-2, or BCL-XL enhances apoptosis in both TP53 / TRP53 wild-type and mutant leukemias and lymphomas. The STING agonist alone, or in combination with the BCL-2 inhibitor venetoclax, was highly effective in AML patient samples, regardless of... TP53 The status remains the same, even in samples that did not show sensitivity to venetoclax alone. In human ENKT lymphoma, the combination of a BCL-XL inhibitor and a STING agonist has the potential to kill malignant cells with nonfunctional TP53 both in vitro and in vivo in xenograft models. ENKT lymphoma and TP53 Mutant AML refers to two blood cancer subtypes of particular concern due to limited treatment options and poor prognosis. Since both BH3 mimics and STING agonists are already in clinical trials (or have been widely approved and used, in the case of venetoclax), there is a possibility of rapidly catching up with currently claimed drug combinations up to clinical trials to achieve immediate translational impact.

[0267] Results - Human AML cell lines Combination therapy of BH3 mimicry drugs and STING agonists was applied to human AML cell lines and TP53-deficient AML cells. Enhanced lethality To determine the potential of STING agonists to enhance the activity of BH3 mimicry drugs, particularly for TP53-deficient hematologic malignancies with poor outcomes after BH3 mimicry drug monotherapy, human AML cell lines were treated in culture with the BH3 mimicry drug ABT-199 / venetoclax or STING agonist compounds 4-46 as a single agent or in combination.

[0268] Figure 7 A-7F shows cell viability assays of MOLM-13 WT, OCI-AML3, THP-1, HL-60, KG-1, and TF-1AML cell lines treated for 48 hours with the BCL-2 inhibitor ABT-199 / venetoclax (thin red line, circle marker) or compound 4-46 (thick blue line, square marker). Data are reported as mean ± SEM.

[0269] Figure 8 and Figure 9 The use of STING agonist compound 4-46 in culture was shown. Figure 8 ) or ABT-199 / Venetok alone ( Figure 9 Cell viability was measured in syngeneic human MOLM-13 AML cells treated for 48 hours with non-targeting sgRNA (thick blue line, square marker), TP53 KO (red dashed line, circle marker), STING KO (green line, triangle marker), and Bak / Bax double KO (purple line, diamond marker). Compounds 4-46 and ABT-199 / venetoclax killed both parental cells and TP53 KO AML cells. STING-deficient cells were completely resistant to compound 4-46 but not to ABT-199 / venetoclax. Bak / Bax double KO AML cells were completely resistant to both agents, confirming the dependence of apoptosis on their individual activity. Figure 10A and 10B The human AML cell line MOLM-13WT was shown to have been treated in culture for 24 hours with a combination of the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM) and the STING agonist compound 4-46 (0-10 µM). Figure 10A ) and OCI-AML3 ( Figure 10B The activity of the cells was measured. This combination therapy enhanced the killing effect of both AML cell types in an additive and synergistic manner. Figure 10C and 10DThe human AML cell line MOLM-13WT was shown to have been treated in culture for 24 hours with a combination of the BCL-2 inhibitor ABT-199 / venetoclax (0-10 µM) and the STING agonist compound 4-46 (0-10 µM). Figure 10C ) and OCI-AML3 ( Figure 10D The heatmap of the activity determination. Figure 10A The data presented in Figure 10C The data presented is the same, and Figure 10D The data presented in Figure 10B The data presented is the same.

[0270] BH3 mimicry and STING agonist synergistically enhance killing in AML patient samples. Next, sample kits of AML patients with different genomic profiles, including those with TP53 deficiency, were examined. Figure 11 ARs show the viability assays of 18 different leukemia blast (LB) cell samples treated with the STING agonist compound 4-46 (thick blue line; circle marker; 0.0001–10 µM), ABT-199 / venetoclax (dashed red line; square marker; 0.0001–10 µM), or a 1:1 combination of the two agents (thin green line; triangle marker; 0.0001–10 µM). Of all the samples tested, 10 / 18 were highly sensitive to compound 4-46 alone, while only 3 / 18 were highly sensitive to ABT-199 / venetoclax. Importantly, 17 / 18 of the AML patient samples showed high sensitivity to the combination of the two drugs. In fact, the patient samples with the lowest sensitivity to ABT-199 / venetoclax were effectively killed in culture by the combination therapy. Therefore, these results demonstrate the potential of combining STING agonists (including any (I)-(V) compounds, such as compounds 4-46) with ABT-199 / venetoclax for the treatment of AML, including cases with TP53 mutations and adverse outcomes.

[0271] Discussion and Conclusion In the examples above, the STING agonist compound 4-46 demonstrated potent cytotoxicity in human AML cell lines when administered as a single agent and in combination with the BCL-2 inhibitor ABT-199 / venetoclax. Loss of STING, achieved through gene knockout in AML cell lines, completely eliminated the effect of compound 4-46 but not the effect of ABT-199 / venetoclax, confirming the dependence of the former compound on the presence of the STING protein. Furthermore, loss of Bax / bak completely eliminated the pro-apoptotic activity of both agents, while TP53 knockout had no effect. AML patient samples with genetic diversity exhibited additive and synergistic cytotoxicity when compound 4-46 was combined with ABT-199 / venetoclax. In fact, patient samples with minimal sensitivity to ABT-199 / venetoclax were potently killed by combination therapy.

[0272] Combination of compound 4-11 and venetoc The combination of the BH3 mimic venetoclax and the STING agonist, compound 4-11, was tested in primary cells using the same procedure described above for the combination of venetoclax and compound 4-46.

[0273] The results of these experiments show that cell viability is demonstrated in Figures 12A-12C In the study, cell viability was demonstrated using compound 4-11 as a monotherapy. Figure 12A Cell viability was demonstrated in the monotherapy of venetoc in [the text abruptly ends here]. Figure 12B Cell viability was demonstrated in the combination therapy of venetotox and compound 4-11. Figure 12C middle.

[0274] Although the present invention has been described with reference to the examples above, it should be understood that the examples are illustrative only and not limited to the invention described herein.

[0275] It will be apparent to those skilled in the art that, although the invention has been described in detail for clarity and understanding, various modifications and alterations can be made to the embodiments and methods described herein without departing from the scope of the inventive concept disclosed herein.

Claims

1. A method for treating a subject's cancer or inhibiting its progression, the method comprising co-administering to the subject: BH3 mimicry drug; and STING agonist.

2. A method for inducing an immune response to cancer in a subject, the method comprising: The subjects were co-administered with a BH3 mimic and a STING agonist, thereby inducing an anticancer immune response in the subjects.

3. The method according to any one of the preceding claims, wherein the cancer comprises cells expressing the STING protein.

4. A method for treating a subject's cancer or inhibiting its progression, the method comprising: The subjects were identified as candidates for BH3 mimicry drug therapy; The cancer was identified as containing cells expressing the STING protein; and The subjects were administered both a BH3 mimic and a STING agonist. The STING agonist and the BH3 mimic drug are administered sequentially or simultaneously.

5. The method according to any one of the preceding claims, wherein the cancer comprises TP53 wild-type cells and TP53 mutant / defective cells.

6. The method according to any one of the preceding claims, wherein the cancer is refractory or resistant to BH3 mimicry drug monotherapy.

7. The method according to any one of the preceding claims, wherein the co-administration comprises the simultaneous administration of the BH3 mimic drug and the STING agonist.

8. The method according to any one of the preceding claims, wherein the co-administration comprises sequentially administering the BH3 mimic drug and the STING agonist.

9. The method according to any one of the preceding claims, wherein the BH3 mimic drug is administered orally or intravenously.

10. The method according to any one of the preceding claims, wherein the STING agonist is administered intravenously or intratumorally.

11. The method according to any one of the preceding claims, wherein the BH3 mimicry and the STING agonist are each administered in a therapeutically effective amount.

12. The method according to any one of the preceding claims, wherein the BH3 mimicry is selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845.

13. The method according to any one of the preceding claims, wherein the STING agonist is selected from any one of the following: compounds 1-1 to 1-172, 2-1 to 2-25, 3-1 to 3-24, 4-1 to 4-112 and 5-1 to 5-15.

14. The method according to any one of the preceding claims, wherein the BH3 mimic drug is administered at an amount of about 50 mg / day to about 600 mg / day.

15. The method according to any one of the preceding claims, wherein the STING agonist is administered at an amount of about 10 µg / week to about 6,400 µg / week over a period of one to three doses every three weeks.

16. The method according to any one of the preceding claims, wherein the cancer is a blood cancer.

17. The method according to any one of the preceding claims, wherein the cancer is selected from the following blood cancers: leukemia, such as acute myeloid leukemia (AML); lymphoma, such as T-cell lymphoma and multiple myeloma (MM); or a relapsed / refractory form of any of these diseases.

18. The method according to any one of the preceding claims, wherein the cancer is a blood cancer selected from acute myeloid leukemia (AML), natural killer / T-cell lymphoma (NKTL), extranodal NK / T-cell lymphoma (ENKTL), and multiple myeloma (MM), or a relapsed / refractory form of any of these diseases.

19. The method according to any one of claims 1 to 16, wherein the cancer is a leukemia selected from: acute myeloid leukemia (AML), including promyelocytic leukemia, chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL), or a relapsed / refractory form of any of these diseases.

20. The method according to any one of claims 1 to 16, wherein the cancer is lymphoma.

21. The method according to any one of claims 1 to 16, wherein the cancer is non-Hodgkin's lymphoma (NHL).

22. The method according to any one of claims 1 to 16, wherein the cancer is selected from the following non-Hodgkin's lymphomas: adult T-cell lymphoma, lymphoblastic lymphoma, peripheral T-cell lymphoma, follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma, natural killer / T-cell lymphoma (NKTL), extranodal NK / T-cell lymphoma (ENKTL), marginal zone lymphoma, Waldenstrom's macroglobulinaemia, and mantle cell lymphoma, or a relapsed / refractory form of any of these diseases.

23. The method according to any one of claims 1 to 15, wherein the cancer is a solid cancer.

24. The method according to any one of claims 1 to 15, wherein the cancer is selected from solid cancers including small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, melanoma, ovarian cancer, neuroblastoma, prostate cancer, and colorectal cancer.

25. The method according to any one of the preceding claims, wherein the subject is a mammal.

26. The method according to any one of the preceding claims, wherein the subject is a human.

27. The method according to any one of the preceding claims, comprising a synergistic combination of co-administering the BH3 mimic drug and the STING agonist.

28. A combination for treating cancer or inhibiting its progression, said combination comprising: BH3 mimicry drug; and STING agonist.

29. The combination according to claim 28, wherein the combination is in the form of a pharmaceutical composition.

30. The combination according to claim 28 or claim 29, wherein the BH3 mimic and the STING agonist are separate formulations.

31. The combination according to any one of claims 28 to 30, wherein the BH3 mimic and the STING agonist are administered simultaneously.

32. The combination according to any one of claims 28 to 30, wherein the BH3 mimic and the STING agonist are administered sequentially.

33. The combination according to any one of claims 28 to 32, wherein the BH3 mimic is selected from BCL-2 selective inhibitors, such as venetoclax; BCL-XL selective inhibitors, such as A-1331852; and MCL-1 selective inhibitors, such as S63845.

34. The combination according to any one of claims 28 to 33, wherein the STING agonist is selected from any one of the following: compounds 1-1 to 1-172, 2-1 to 2-25, 3-1 to 3-24, 4-1 to 4-112 and 5-1 to 5-15.

35. The combination of any one of claims 28 to 34, wherein the cancer comprises cells expressing the STING protein.

36. The combination of any one of claims 28 to 35, wherein the cancer comprises TP53 wild-type cells and TP53 mutant cells.

37. The combination of any one of claims 28 to 36, wherein the cancer is refractory or resistant to BH3 mimicry drug monotherapy.

38. The combination of any one of claims 28 to 37, wherein the combination is a synergistic combination of the BH3 mimicry and the STING agonist.

39. Use of a BH3 mimicry and a STING agonist for the preparation of a medicament for treating cancer in a subject, wherein the BH3 mimicry and the STING agonist are administered together.

40. The use according to claim 39, wherein the co-application is applied sequentially or simultaneously.

41. Use of a STING agonist for the preparation of a medicine for use in combination with a BH3 mimicry drug to treat cancer in a subject.

42. Use of a BH3 mimicry for the preparation of a medicine for use in combination with a STING agonist to treat cancer in a subject.

43. The use according to claim 41 or claim 42, wherein the combined application is applied sequentially or simultaneously.

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