Combination Therapy of BCL2 Inhibitors and Peptides for Treating Proliferative Diseases - Patent application

Abstract

The present disclosure relates to combination therapies comprising Bcl2 inhibitors and peptides. The present disclosure further relates to compositions, methods and kits comprising Bcl2 inhibitors and peptides for the treatment of proliferative disorders.

Description

[Technical field]

[0001] The present disclosure relates to combination therapy of B-cell lymphoma 2 (Bcl2) inhibitors and peptides for the treatment of proliferative diseases. [Background technology]

[0002] References that are considered relevant as background to the presently disclosed subject matter are listed below. -[1]Lessene G, Czabotar PE, Colman PM.BCL-2 family antagonists for cancer therapy.Nat Rev Drug Discov.2008;7(12):989-1000. -[2] International Publication No. 2006 / 046239 -[3] International Publication No. 2007 / 122622 -[4] International Publication No. 2007 / 091240 -[5] International Publication No. 2008 / 075349 -[6]Sandler, U. et al., 2010, Recent advances in clinical medicine, ISSN:1790-5125. -[7]Sandler, U. et al., 2010, J Experimental Therapeutics and Oncology 8:327-339. -[8] International Publication No. 2015 / 083167 -[9] International Publication No. 2017 / 134668

[0003] The acknowledgment of the above references herein should not be inferred as meaning that they are in any way relevant to the patentability of the presently disclosed subject matter.

[0004] 2. Background of the Invention The BCL2-related proteins are a large family of proteins that operate through nonenzymatic protein interactions that regulate the intrinsic or mitochondrial pathways to apoptosis, protect against apoptosis, or promote apoptosis.

[0005] Pro-survival proteins include, for example, BCL-2 itself, B-cell lymphoma extra-large (BCL-xL), myeloid cell leukemia-1 (MCL-1), B-cell lymphoma-w (BCL-w), and BCL-2-related gene 1 expressed in fetal liver.

[0006] On the other hand, pro-apoptotic proteins are further classified into (a) pro-apoptotic BH3-only proteins, including BCL-2 interacting mediator of cell death (BIM), p53 upregulated modulator of apoptosis, and BCL-2-associated death promoter, and (b) pro-apoptotic effector proteins BCL-2-associated X protein (BAX) and BCL-2 homologous antagonist killer (BAK).

[0007] The balance of activity between pro-survival and BH3-only pro-apoptotic proteins determines whether a cell will survive or undergo apoptosis, and collectively they act to integrate diverse extracellular and intracellular signals that promote survival (e.g., growth factors, nutrients) or stress-induced death (e.g., oncogenic / proliferative, DNA damage, etc.).

[0008] BH3 mimetics are a new class of anticancer drugs that mimic the action of BH3-only proteins in that they bind to pro-survival proteins such as BCL2 in the same manner as BH3-only proteins and inhibit the ability of BCL2 to bind to BAX or BAK [1]. Because BCL2 also exists bound to natural BH3-only proteins, BH3 mimetics can also surrogate for such endogenous activators of apoptosis. For example, venetoclax is a BCL2-selective BH3 mimetic, and its in vitro addition to BCL2-overexpressing cancer cells potently induces apoptosis.

[0009] A peptide called "T101" encoded by a cDNA unique to the human thymus has previously been identified. This peptide and its derivatives have been implicated in cancer therapy, inter alia, through the role of T101 as a stimulator of the immune system (WO 2006 / 046239, [2]). WO 2006 / 046239 demonstrates that T101 can stimulate the immune system and reduce tumor size, suggesting that the peptide influences cancer cell proliferation. WO 2006 / 046239 also suggests an immune-based role for T101, for example, in protecting patients during standard chemotherapy.

[0010] The treatment of cancer by using T101 has also been suggested in WO 2007 / 122622 [3], which demonstrates, inter alia, the effect of T101 on the development of various types of tumors. Peptide T101 was also described in WO 2007 / 091240 [4] for the treatment of immune disorders, as well as in WO 2008 / 075349 [5] and in publications by Sandler et al. [6-7] for the treatment or prevention of disorders involving cells bearing the T1 / ST2 receptor.

[0011] In addition, a peptide derivative of T101 called "Nerofe" has been reported to reduce the secretion of proteins known to be associated with cancer metastasis by cancer cells in vitro and to directly inhibit cancer cell migration. Furthermore, the peptide has been shown to affect serum levels of vascular endothelial growth factor (VEGF) in cancer patients (WO 2015 / 083167, [8]), suggesting its use in methods of preventing or treating cancer metastasis.

[0012] Furthermore, the above-mentioned peptide, called “Nerofe”, was also shown in WO 2017 / 134668[9] to be involved in the induction of ER stress, which contributes to the promotion of cell death, and was suggested to reduce the standard therapeutic dose of administered anticancer drugs in treated cancer patients. Summary of the Invention

[0013] In a first aspect, the present disclosure provides a combination composition comprising at least one inhibitor against B-cell lymphoma 2 (Bcl2) pro-survival protein, an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier, diluent, excipient and / or additive.

[0014] In a second aspect, the disclosure provides a method for achieving treatment, prevention, amelioration, reduction, or delayed onset of at least one proliferative disorder in a subject in need thereof, comprising administering a therapeutically effective amount of at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising an amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharmaceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0015] In a third aspect, the present disclosure provides at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, for use in a method for treating, preventing, ameliorating, alleviating, or delaying the onset of at least one proliferative disorder.

[0016] In a fourth aspect, the present disclosure provides a method for producing a pharmaceutical composition comprising: (a) at least one inhibitor against Bcl2 pro-survival protein, or any pharma- ceutically acceptable salt thereof, optionally in a first dosage form; (b) an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, which may be in a second dosage form; A kit comprising:

[0017] In a fifth aspect, the present disclosure provides a method for achieving treatment, prevention, amelioration, alleviation, or delay in onset of at least one proliferative disorder in a subject treated with an inhibitor against at least one Bcl2 pro-survival protein, the method comprising administering to the subject a therapeutically effective amount of an isolated peptide comprising an amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0018] In a sixth aspect, the present disclosure provides a method for achieving treatment, prevention, amelioration, reduction, or delay in onset of at least one proliferative disorder in a subject treated with at least one isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, comprising administering to the subject a therapeutically effective amount of at least one inhibitor against Bcl2 pro-survival protein, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0019] In order to better understand the subject matter disclosed herein, and to illustrate how it may be carried out in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0020] [Figure 1] 1 is a graph showing the synergy between a drug called Nerofe and ABT-737 in inducing apoptosis in U937 cells, as measured via activation of caspase 3. [Diagram 2] 1 is a graph showing that Nerofe synergizes with ABT-737 to induce death of U937 cells as measured by percentage of cell viability using resazurin. [Diagram 3] 1 is a graph showing that Nerofe does not synergize with azacytidine to induce death of U937 cells as measured by percentage of cell viability using resazurin. [Figure 4] 1 is a graph showing that Nerofe synergizes with ABT-737 to reduce the viability of CT26 cells as measured by the percentage of cell viability using resazurin. [Diagram 5] Photograph of a Western blot assay staining cleaved caspase 3 in CT26 cells treated with Nerofe in the presence / absence of ABT-737. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] A new class of anti-cancer drugs, Bcl2 pro-survival protein inhibitors, are shown herein to act synergistically with a peptide called "Nerofe" in inducing apoptosis and cell death in malignant cells.

[0022] Thus, in a first aspect, the present disclosure provides a combination composition comprising at least one inhibitor against B-cell lymphoma 2 (Bcl2) pro-survival protein, an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier, diluent, excipient and / or additive.

[0023] The term "isolated peptide" as defined herein includes the isolated peptide comprising the amino acid sequence set forth in SEQ ID NO:1 (i.e., the amino acid sequence Trp Trp Thr Phe Phe Leu Pro Ser Thr Leu Trp Glu Arg Lys, all in the D-form), referred to herein as "dTCApF" or "Nerofe", and a functional derivative of the amino acid sequence set forth in SEQ ID NO:1, or a pharma- ceutically acceptable salt of said isolated peptide. Any pharma- ceutically acceptable salt of the isolated peptide as defined herein, particularly the acetate salt of the peptide, is included in the present disclosure.

[0024] The term "peptide" as defined herein refers to a molecular chain of amino acid residues, each of which may be optionally modified, for example, by manosylation, glycosylation, amidation (e.g., C-terminal amide), carboxylation or phosphorylation. Peptides may be obtained synthetically, by genetic engineering techniques, by expression in a host cell, or by any other suitable means. Methods for producing peptides are well known in the art.

[0025] More specifically, an "amino acid molecule", "amino acid sequence" or "peptide sequence" is the order in which amino acid residues connected by peptide bonds occur in a chain of peptides and proteins. The sequence is generally passed down in the order from the N-terminus, which contains a free amino group, to the C-terminus, which contains the amide. Although an amino acid sequence is often called a peptide or protein sequence when it represents the primary structure of a protein, a distinction must be made between the term "amino acid sequence" or "peptide sequence" and the term "protein", since a protein is defined as an amino acid sequence that has been folded into a specific three-dimensional configuration and has typically undergone post-translational modifications such as phosphorylation, acetylation, glycosylation, manosylation, amidation, carboxylation, sulfhydryl bond formation, cleavage, etc.

[0026] Amino acid, as used herein, refers to natural and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to natural amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are later modified (e.g., hydroxyproline, y-carboxyglutamic acid, and O-phosphoserine). "Amino acid analog" refers to a compound that has the same basic chemical structure as a natural amino acid, i.e., an alpha carbon attached to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Such analogs have modified R groups or modified peptide backbones, but retain the same basic chemical structure as a natural amino acid. "Amino acid mimetics" refers to a compound that has a structure that differs from the general chemical structure of an amino acid, but functions similarly to a natural amino acid. Amino acids may be referred to herein by either their commonly known three letter symbols or the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0027] The term "isolated" refers to molecules, such as amino acid sequences or peptides, that are removed and isolated or separated from their natural environment.

[0028] As used herein, the term "amino acid" refers to naturally occurring and synthetic amino acid residues, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine.

[0029] The term amino acid encompasses L-amino acids and D-amino acids, which are mirror images of L-amino acids with the chirality at the carbon alpha reversed. D-amino acids are highly resistant to protease-mediated degradation and have a low immunogenic response.

[0030] The term "amino acid sequence" or "peptide sequence" also refers to the order in which amino acid residues linked by peptide bonds occur in a chain of peptides and proteins. The sequence is generally given in order from the N-terminus, which contains a free amino group, to the C-terminus, which contains a free carboxyl group.

[0031] The term "comprising" means that the isolated peptide according to the present disclosure includes the peptide set forth by SEQ ID NO:1, but may also include additional amino acid residues at the N-terminus or C-terminus, or both, of the peptide.

[0032] As noted above, the present disclosure also encompasses isolated peptides, including derivatives of peptides having the amino acid sequence set forth in SEQ ID NO: 1. The term "derivative(s)" is meant to include peptides that contain the amino acid sequence set forth in SEQ ID NO: 1, but have one or more amino acid differences in their overall sequence, i.e., deletions, substitutions (e.g., replacement of at least one amino acid with another amino acid), inversions or additions within the overall sequence of SEQ ID NO: 1. The term also encompasses the replacement of at least one amino acid residue in the overall sequence with its respective L-amino acid residue.

[0033] In certain embodiments, the modified isolated peptide of the disclosure or composition of the invention has at least 70%, or 80%, or 90%, or 95%, or particularly 99% identity to the sequence corresponding to SEQ ID NO:1.

[0034] In some embodiments, a peptide comprising the amino acid sequence set forth in SEQ ID NO:1 may contain one or more amino acid residues substituted by conservative substitutions without significantly affecting the biological characteristics of the modified peptide compared to an unmodified peptide having the amino acid sequence of SEQ ID NO:1.

[0035] An amino acid "substitution" is the result of replacing one amino acid with another amino acid having similar structural and / or chemical properties, i.e., a conservative amino acid substitution. Amino acid substitutions can be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or the amphipathic nature of the residues involved. For example, each of the following eight groups contains amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G), 2) Aspartic acid (D), glutamic acid (E), 3) Asparagine (N), Glutamine (Q), 4) Arginine (R), Lysine (K), 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V), 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W), 7) Serine (S), Threonine (T), and 8) Cysteine ​​(C), Methionine (M).

[0036] It should be understood that these peptide derivatives should not change the biological activity of the original peptide. The term "functional" means that the modified peptide (i.e., the derivative) retains a biological activity qualitatively similar to that of the unmodified peptide. The biological activity of the derivative can be determined by monitoring the effect of the derivative upon administration to an animal model as described herein, i.e., as known in the art.

[0037] In certain embodiments, the present disclosure relates to a functional derivative or fragment of an isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1, which functional derivative or fragment has an amino acid sequence that is at least about 70%, 75%, 80%, 85%, 90%, or 95%, particularly 99% identical to the amino acid sequence of the unmodified isolated polypeptide of the present disclosure, i.e., the amino acid sequence set forth in SEQ ID NO:1, and retains biological activity qualitatively similar to that of the unmodified peptide.

[0038] In some specific embodiments, the isolated peptide consists of the amino acid sequence of SEQ ID NO:1.

[0039] Still further, as used herein, the term Bcl-2 pro-survival protein refers to a proto-oncogene protein known as an apoptosis inhibitor. Bcl-2 proteins form the basis of a growing family of related proteins collectively referred to herein as the Bcl-2 family of proteins. These proteins are known to control apoptotic cell death via the mitochondrial pathway.

[0040] As is understood in the art, members of the Bcl-2 family are either pro-survival or pro-apoptotic, but regardless of their activity, they all share significant sequence and structural homology. Specifically, the Bcl-2 family of proteins is characterized by up to four regions of sequence homology known as Bcl-2 homology (BH) domains.

[0041] The Bcl-2 family of proteins comprises three distinct groups of proteins, the first group being the pro-survival or anti-apoptotic group, referred to herein as the "Bcl-2 pro-survival proteins", the second group being the pro-apoptotic group, which includes BAX and BAK, and the third group being referred to herein as the BH3-only proteins that exhibit pro-apoptotic activity.

[0042] "Bcl-2 pro-survival proteins" or "anti-apoptotic" or "Bcl-2-like" as used herein refer to a group of proteins that play a role in protecting cells from apoptotic stimuli and are characterized sequentially by containing all four BH domains.

[0043] It has previously been shown that the balance of pro-survival and pro-apoptotic Bcl-2 proteins, as well as the specific interactions between them, determine the activity of the protein family in response to apoptotic stimuli. For example, in contrast to the Bcl-2 pro-survival proteins, BH3-only proteins initiate apoptosis in response to a variety of cellular stresses, including DNA damage, growth factor deprivation, and endoplasmic reticulum stress. Activation of BH3-only proteins may involve transcriptional upregulation or modification.

[0044] Several mechanisms have been suggested in the art to explain the diverse activities and tight regulation of apoptosis in the Bcl-2 family, some of which involve Bax and Bak. Regardless of the exact means, it has become clear that the balance between pro-survival and pro-apoptotic (BH3-only) proteins is key to this activation.

[0045] According to some embodiments, the Bcl2 inhibitor may antagonize any Bcl-2 pro-survival protein, such as at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1, and Bcl-B / Bcl2L10.

[0046] In some further embodiments, the Bcl-2 pro-survival protein of the compositions of the invention is at least one of Bcl-2, B-cell lymphoma extra-large (Bcl-xL), myeloid cell leukemia-1 (Mcl-1), B-cell lymphoma-w (Bcl-w), B-cell lymphoma 2-associated protein A1 (Bcl2A1), and B-cell lymphoma 2-like 10 (Bcl-B / Bcl2L10).

[0047] More specifically, in some embodiments, the Bcl2 inhibitor may bind and antagonize the Bcl-2 protein. Bcl-2 (B-cell CLL / lymphoma 2), as used herein, is an integral outer mitochondrial membrane protein that blocks apoptotic death of some cells, such as lymphocytes. Bcl-2 suppresses apoptosis in various cell systems, including factor-dependent lymphohematopoietic cells and neuronal cells. It regulates cell death by controlling mitochondrial membrane permeability. Bcl-2 appears to function in a feedback loop system with caspases, inhibiting caspase activity by preventing the release of cytochrome c from mitochondria and / or by binding to apoptosis activating factor (APAF-1). It is noted that in certain embodiments, the present invention refers to the human Bcl-2 protein represented by GenBank Accession Nos. NP_000624 and SEQ ID NO: 2, and NP_000648, SEQ ID NO: 3, which is encoded by the Bcl-2 genes of GenBank Accession Nos. NM_000633, SEQ ID NO: 4, and NM_000657, SEQ ID NO: 5.

[0048] In yet another embodiment, the Bcl2 inhibitor can bind and antagonize Bcl-xL. As used herein, B-cell lymphoma extra-large (Bcl-xL) is a transmembrane molecule in mitochondria. It is a member of the Bcl-2 family of proteins and acts as a pro-survival protein by preventing the release of mitochondrial contents such as cytochrome c, which leads to caspase activation. In a particular embodiment, the present invention relates to the human Bcl-xL protein (GenBank Accession No. CAA80661, SEQ ID NO: 6), which is encoded by the Bcl-xL gene represented by GenBank Accession No. Z23115 and SEQ ID NO: 7.

[0049] In yet another embodiment, the Bcl2 inhibitor may bind to and antagonize any one of the human Bcl-2 pro-survival proteins, Mcl-1, Bcl-w, Bcl2A1 and Bcl-B / Bcl2L10, which are designated by Accession Nos. NP_068779.1, AAB09055, NP_004040.1 and NP_001293097.1, respectively.

[0050] In some other embodiments, the at least one inhibitor against Bcl2 pro-survival protein is at least one Bcl-2 homology 3 (BH3) mimetic compound.

[0051] As mentioned above, BH3 mimetics are a class of anti-cancer drugs that mimic the action of BH3-only proteins in that they bind to pro-survival proteins such as BCL2 in the same way and inhibit the ability of BCL2 to bind to BAX or BAK.

[0052] In some particular embodiments, the BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-2-yl]methyl]piperazin-1-yl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohexenyl)methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), hex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy -2H-pyrrol-5-yl)-1H-indole (GX15-070), or 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852), or a combination thereof.

[0053] In some further embodiments, the at least one inhibitor or BH3 mimetic compound against Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0054] ABT-737 is a small molecule drug that inhibits Bcl-2 and Bcl-xL, two members of the evolutionarily conserved Bcl-2 family of proteins that share a Bcl-2 homology (BH) domain. ABT-737 is not bioavailable following oral administration, leading to navitoclax (ABT-263), an orally available derivative with similar activity against small cell lung cancer (SCLC) cell lines.

[0055] ABT-737 has the following chemical structure, represented by Formula I:

[0056] [ka]

[0057] The systematic name of ABT-737 (TUPAC) is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (C 42 H 45 CIn6O5S2; CAS number: 852808-04-9). The molecular weight of ABT-737 is 813.43 g / mol.

[0058] In some further embodiments, the at least one inhibitor or BH3 mimetic compound against Bcl2 pro-survival protein is 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0059] ABT-199 or venetoclax, sold under the trade names Venclexta and Venclyxto, is a drug used to treat adults with chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), or acute myeloid leukemia (AML). Venetoclax binds to the Bcl-2 protein, which is found in high amounts in CLL cancer cells, where it helps the cells survive longer in the body and makes them resistant to cancer drugs. By binding to and blocking the action of Bcl-2, venetoclax causes the death of cancer cells, thereby slowing the progression of the disease. Venetoclax is the first selective BCL2 inhibitor to be approved for routine clinical practice, specifically in chronic lymphocytic leukemia (CLL) and acute myeloid leukemia (AML).

[0060] ABT-199 has the following chemical structure, represented by Formula II:

[0061] [ka]

[0062] The systematic name (IUPAC) of ABT-199 is 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)benzamide (CHCNOS; CAS number: 1257044-40-8). The molecular weight of ABT-199 is 868.45 g / mol.

[0063] In some further embodiments, the at least one inhibitor of Bcl2 pro-survival protein or BH3 mimetic compound is 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)-benzamide (ABT-263), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0064] ABT-263 or navitoclax is an orally active anticancer drug that is a Bcl-2 inhibitor. ABT-263 has the following chemical structure, represented by Formula III:

[0065] [ka]

[0066] The systematic name (IUPAC) of ABT-263 is 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (C 47 H 55 ClF3N5O6S3; CAS number: 923564-51-6). The molecular weight of ABT-263 is 974.61 g / mol.

[0067] In some further embodiments, the at least one inhibitor or BH3 mimetic compound against Bcl2 pro-survival protein is 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)-1H-indole (GX15-070), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0068] GX15-070 or obatoclax is a drug for the treatment of various types of cancer. Obatoclax is an inhibitor of the Bcl-2 family of proteins. This inhibition induces apoptosis in cancer cells and prevents tumor growth. GX15-070 has the following chemical structure, represented by formula IV:

[0069] [ka]

[0070] The systematic name (IUPAC) of GX15-070 is 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)-1H-indole (C 20 H 19 N3O; CAS number: 803712-7-6). The molecular weight of GX15-070 is 317.392 g / mol.

[0071] In some further embodiments, the at least one inhibitor or BH3 mimetic compound against Bcl2 pro-survival protein is 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0072] A-1331852 is a first-in-class, orally active BCL-XL inhibitor that selectively and potently induces apoptosis in BCL-XL-dependent tumor cells. A-1331852 has the following chemical structure, represented by Formula V:

[0073] [ka]

[0074] The systematic name (IUPAC) of A-1331852 is 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (C 38 H 38 N6O3S; CAS number: 1430844-80-6). The molecular weight of A-1331852 is 658.27 g / mol.

[0075] In some embodiments, the composition may include at least one additional therapeutic agent, such as an anti-cancer agent. In some further embodiments, the composition does not include taxol, also known as paclitaxel, doxorubicin, or an anti-PDL1 antibody.

[0076] In yet some further embodiments, the compositions of the present disclosure are for use in a method for treating, preventing, ameliorating, reducing, or delaying the onset of at least one proliferative disorder.

[0077] In some embodiments, the proliferative disorder associated with the compositions of the invention is associated with Bcl2 overexpression.

[0078] In some further embodiments, the proliferative disorder is at least one hematological cancer or neoplasm.

[0079] In some specific embodiments, the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0080] As shown in the Examples below, the combination compositions were shown to act synergistically to induce apoptosis and cell death in U937 cells and CT26 cells, cell lines derived from histiocytic lymphoma and colorectal carcinoma, respectively.

[0081] In some specific embodiments, the proliferative disorder is at least one of histiocytic lymphoma or colorectal cancer.

[0082] It should be noted that the combination composition of the present invention can induce or enhance apoptosis of malignant / cancer cells. More specifically, the combination composition of the present invention can result in an increase, enhancement, induction or elevation of apoptosis of treated cells, and the increase, induction or elevation of apoptosis can be about 1% to 99.9%, specifically about 1% to about 95%, about 5% to 90%, about 10% to 85%, about 15% to 80%, about 20% to 75%, about 25% to 70%, about 30% to 65%, about 35% to 60%, about 40% to 55%, about 45% to 50%. More specifically, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99.9%. More specifically, an increase of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% compared to an untreated control.

[0083] With regard to the above, it should be understood that where provided, percentage values ​​such as 10%, 50%, 120%, 500%, etc. are interchangeable with "fold change" values, i.e. 0.1, 0.5, 1.2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, etc., respectively.

[0084] The term "apoptosis" refers to a network of regulated biochemical events that result in a selective form of cell suicide and are characterized by readily observable morphological and biochemical phenomena. Cells undergoing apoptosis display characteristic morphological and biochemical features. These features include chromatin clumping or condensation, DNA fragmentation, nuclear and cytoplasmic condensation, and partitioning of the cytoplasm and nucleus into membrane-bound vesicles containing ribosomes (apoptotic bodies), morphologically intact mitochondria, and nuclear material. Cytochrome C release from mitochondria is seen as an indicator of mitochondrial dysfunction associated with apoptosis.

[0085] As mentioned above, apoptosis is a tightly regulated form of active cell death that is necessary for development and biological homeostasis. Death via the apoptotic pathway is achieved, inter alia, by the activation of a family of highly potent and specific proteases called caspases (of cysteine-aspartic acid proteases).

[0086] The activity of caspases is tightly regulated, and cells maintain several "checkpoints" to control their activity. The first level of regulation is intrinsic to the caspases themselves: caspases are initially transcribed as weakly active zymogens and are cleaved to form active enzymes only upon appropriate stimuli. The second level of caspase regulation is achieved by inhibitors, namely a family of proteins called IAPs (inhibitors of apoptosis proteins).

[0087] Thus, in a second aspect, the present disclosure provides a method for achieving treatment, prevention, amelioration, alleviation, or delay in onset of at least one proliferative disorder in a subject in need thereof, comprising administering a therapeutically effective amount of at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0088] In some embodiments, a modified isolated peptide suitable for the methods of the present disclosure or invention has at least 70%, or 80%, or 90%, or 95% identity to a sequence corresponding to SEQ ID NO:1.

[0089] In some specific embodiments, the isolated peptide consists of the amino acid sequence of SEQ ID NO:1.

[0090] In another embodiment, the Bcl-2 pro-survival protein relevant to the methods of the invention is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1 and Bcl-B / Bcl2L10.

[0091] In some further embodiments, the at least one inhibitor against Bcl2 pro-survival protein suitable for the methods of the invention is at least one BH3 mimetic compound.

[0092] In some further specific embodiments, the BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4 -chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 4-(4-{[2-(4-chlorophenyl)-5,5- Dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene )-3-methoxy-2H-pyrrol-5-yl)-1H-indole (GX15-070), or 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).

[0093] In yet another specific embodiment, the at least one inhibitor against Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0094] As used herein, a "proliferative disorder" is a disorder that exhibits hyperproliferation. The term refers to cell division and growth that is not part of the normal cell turnover, metabolism, growth, or proliferation of the whole organism. Unwanted proliferation of cells is found in tumors and other pathological proliferations of cells that do not perform normal functions and continue unchecked, for the most part, at a proliferation rate that exceeds that of cells of normal tissues in the absence of external intervention. Pathological conditions that arise due to unwanted proliferation of cells are referred to herein as "hyperproliferative diseases" or "hyperproliferative disorders." It should be noted that the terms "proliferative disorder," "cancer," "tumor," and "malignant tumor" all relate equally to hyperplasia of tissues or organs. In general, the compositions and methods of the present invention can be used to treat non-solid and solid tumors.

[0095] The malignant tumor contemplated in the present invention may be any one of lymphoma, leukemia, carcinoma, melanoma, myeloma and sarcoma.

[0096] Lymphoma is a cancer in the lymphatic cells of the immune system. Typically, lymphomas exist as solid tumors of lymphatic cells. These malignant cells often originate from lymph nodes and appear as lymph node enlargements (tumors). It may also affect other organs, in which case it is called extranodal lymphoma. Non-limiting examples of lymphoma include Hodgkin's disease, non-Hodgkin's lymphoma, and Burkitt's lymphoma.

[0097] Leukemia refers to a progressive malignant disease of the blood-forming organs and is generally characterized by distorted proliferation and development of white blood cells and their precursors in the blood and bone marrow. Leukemias are generally classified according to (1) the duration and character of the disease - acute or chronic; (2) the type of cells involved - myeloid (myeloid), lymphocytic (lymphoid), or monocytic; and (3) the increase or absence of an increase in the number of abnormal cells in the blood - leukemia or non-leukemia (subleukemia).

[0098] Carcinoma, as used herein, refers to an invasive malignant tumor composed of transformed epithelial cells. Alternatively, it refers to a malignant tumor composed of transformed cells of unknown histogenesis, but which has specific molecular or histological characteristics associated with epithelial cells (e.g., production of cytokeratins or intercellular bridges).

[0099] Melanoma, as used herein, is a malignant tumor of melanocytes. Melanocytes are the cells that produce melanin, the dark pigment responsible for skin color. They are found primarily in the skin, but are also found in other parts of the body, including the intestine and eyes. Melanoma can occur in any part of the body that contains melanocytes.

[0100] Sarcomas are cancers that arise from transformed connective tissue cells. These cells originate from the embryonic mesoderm or intermediate layer that forms bone, cartilage, and fat tissue. This contrasts with carcinomas that are derived from the epithelium, which lines the surfaces of structures throughout the body and is the origin of cancers in the breast, colon, and pancreas.

[0101] Myeloma, as referred to herein, is a cancer of plasma cells, a type of white blood cell normally involved in the production of antibodies. Collections of abnormal cells accumulate in the bones, where they cause bone lesions, and in the bone marrow, where they interfere with the production of normal blood cells. Most cases of myeloma are also characterized by the production of paraproteins, abnormal antibodies that can cause kidney problems and interfere with normal antibody production, resulting in immune deficiency. Hypercalcemia (high calcium levels) is often encountered.

[0102] Additional malignancies that may find utility in the present invention include, but are not limited to, hematological malignancies (including lymphomas, leukemias, and myeloproliferative disorders), aplastic and aplastic anemias (both virally induced and idiopathic), myelodysplastic syndromes, paraneoplastic syndromes of all types (both immune-mediated and idiopathic), and solid tumors (GI tract, colon, lung, liver, breast, prostate, pancreas, and Kaposi's sarcoma. More specifically, the malignant disorder may be lymphoma. Non-limiting examples of cancers treatable according to the present invention include hematopoietic malignancies, e.g. For example, all types of lymphoma, leukemias, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), mutant KRAS tumors, chronic myeloid leukemia (CML), myelodysplastic syndromes (MDS), mast cell leukemia, hairy cell leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, Burkitt's lymphoma, and multiple myeloma, as well as solid tumors, such as tumors of the lips and oral cavity, pharynx, larynx, paranasal sinuses, major salivary glands, thyroid, esophagus, stomach, small intestine, colon, colorectum, anal canal, liver, gallbladder, extrahepatic bile duct, and the like. tumors of the following organs: bile ducts, ampulla of Vater, exocrine pancreas, lung, pleural mesothelioma, bone, soft tissue sarcoma, skin, breast, vulva, vagina, cervix, uterus, ovary, fallopian tube, gestational chorionic neoplasm, penis, prostate, testicle, kidney, renal pelvis, ureter, bladder, urethra, carcinoma of the eyelid, carcinoma of the conjunctiva, malignant melanoma of the conjunctiva, malignant melanoma of the uvea, retinoblastoma, carcinoma of the lacrimal gland, sarcoma of the orbit, brain, spinal cord, vasculature, angiosarcoma, and Kaposi's sarcoma.

[0103] Specifically, the anti-apoptotic protein Bcl-2 governs pro-survival pathways and has been shown to be overexpressed in a variety of tumor types, including small cell lung cancer, melanoma, prostate cancer, and breast cancer.

[0104] In some embodiments, the proliferative disorder relevant to the methods of the invention is associated with Bcl2 overexpression or a Bcl-2 overexpression disorder or a Bcl-2 mediated disorder.

[0105] The terms "Bcl2 overexpression associated disorder" or "Bcl-2 overexpression disorder" and "Bcl-2 mediated disorder" refer to pathological and disease conditions in which the Bcl-2 protein is overexpressed, as set forth herein above. Furthermore, the term also encompasses conditions in which Bcl-2 plays a role. Such a role may be directly related to the pathological condition or indirectly related to the condition. A common feature of this class of conditions is that they may be ameliorated by inhibiting the expression, activity, function, or association with the Bcl-2 protein.

[0106] The term "overexpressed" refers to an increase in the measurable expression level of the Bcl-2 gene as measured by the amount of RNA and / or protein in a sample compared to the measurable expression level of the Bcl-2 gene in a second sample, specifically a control sample. "Overexpressed Bcl-2" can be measured and evaluated using the ratio of the expression level of Bcl-2 in a sample compared to the average expression level of Bcl-2 in a control sample, which is not equal, specifically greater than 1.0. When determining overexpression based on a ratio, the RNA or protein is overexpressed when the ratio of the expression level in a first sample compared to a second sample is greater than 1.0. For example, a ratio of greater than 1.2, 1.5, 1.7, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more.

[0107] Thus, a Bcl-2 overexpressing pathological disorder means a disorder characterized by overexpression of Bcl-2 in a subject, or in an affected tissue of a subject, compared to a healthy subject or healthy tissue of the same subject.

[0108] It should be noted that Bcl-2 overexpression disorders can be caused by chromosomal translocations, hypomethylation and downregulation of microRNAs that target Bcl-2.

[0109] B-cell lymphoma 2 (BCL2) protein is variably highly expressed in many hematological malignancies and is known to provide protection from cell death induced by oncogenic and external stresses. For example, venetoclax is the first selective BCL2 inhibitor and the first in a class of anticancer drugs (BH3 mimetics) approved for routine clinical practice in chronic lymphocytic leukemia (CLL) and acute myeloid leukemia (AML).

[0110] In certain embodiments, the compositions or methods of the invention are for treating, preventing, ameliorating, ameliorating, or delaying the onset of chronic lymphocytic leukemia (CLL), a type of cancer in which the bone marrow produces too many lymphocytes.

[0111] In some specific embodiments, the compositions or methods of the invention are for treating, preventing, ameliorating, mitigating, or delaying the onset of acute myeloid leukemia (AML), a cancer of the myeloid system of blood cells characterized by the rapid proliferation of abnormal cells that accumulate in the bone marrow and blood and interfere with normal blood cell production.

[0112] In some specific embodiments, the compositions or methods of the present invention are for treating, preventing, ameliorating, alleviating, or delaying the onset of mutant KRAS tumors. Mutant KRAS tumors or KRAS-driven tumors refer to a type of cancer tumor in which the Kirsten rat sarcoma viral oncogene (KRAS) is mutated. The KRAS oncogene has the highest mutation rate among all cancers and is associated with a lineage of highly lethal cancers, such as pancreatic ductal adenocarcinoma (PDAC), non-small cell lung cancer (NSCLC), and colorectal cancer (CRC).

[0113] Thus, in some specific embodiments, the proliferative disorder is at least one hematological cancer or neoplasm.

[0114] In further specific embodiments, the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0115] In certain specific embodiments, the proliferations associated with the methods of the invention are histiocytic lymphoma and colorectal cancer.

[0116] In some further embodiments, the methods of the present invention enhance or induce apoptosis in malignant / cancerous cells in a subject.

[0117] In yet another embodiment, the methods of the invention further comprise administering at least one therapeutic agent, for example at least one additional anti-cancer agent.

[0118] In some specific embodiments, the additional anti-cancer agent is not taxol or doxorubicin or an anti-PDL1 antibody.

[0119] In some embodiments of the methods of the invention, at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 are administered simultaneously or sequentially.

[0120] In a third aspect, the present disclosure provides at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, for use in a method for treating, preventing, ameliorating, alleviating, or delaying the onset of at least one proliferative disorder.

[0121] In some embodiments, a modified isolated peptide suitable for use with the present disclosure or invention has at least 70%, or 80%, or 90%, or 95% identity to a sequence corresponding to SEQ ID NO:1.

[0122] In some specific embodiments, the isolated peptide consists of the amino acid sequence of SEQ ID NO:1.

[0123] In another embodiment, the Bcl-2 pro-survival protein relevant for use according to the invention is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1 and Bcl-B / Bcl2L10.

[0124] In some further embodiments, the at least one inhibitor against Bcl2 pro-survival protein suitable for use according to the present invention is at least one BH3 mimetic compound.

[0125] In some further specific embodiments, the BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4 -chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 4-(4-{[2-(4-chlorophenyl)-5,5- Dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene )-3-methoxy-2H-pyrrol-5-yl)-1H-indole (GX15-070), or 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).

[0126] In yet another specific embodiment, the at least one inhibitor against Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0127] In some embodiments, at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 for use according to the present invention are administered simultaneously or sequentially.

[0128] In yet another embodiment, the use of at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 according to the present invention further comprises administering at least one additional anti-cancer agent.

[0129] In some specific embodiments, the additional anti-cancer agent is not taxol or doxorubicin or an anti-PDL1 antibody.

[0130] In some embodiments, the proliferative disorder relevant for use according to the invention is associated with Bcl2 overexpression.

[0131] In some specific embodiments, the proliferative disorder is at least one hematological cancer or neoplasm.

[0132] In further specific embodiments, the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0133] In some specific embodiments, the proliferations suitable for use with the present invention are histiocytic lymphoma and colorectal cancer, respectively.

[0134] In some embodiments, the isolated peptides defined herein and / or suitable for the methods and uses disclosed herein, or the Bcl2 inhibitors or pharma- ceutically acceptable salts thereof, may be included in a pharmaceutical composition. The term "pharmaceutical composition" as defined herein refers to the isolated peptides and / or Bcl2 inhibitors and, optionally, at least one pharma- ceutically acceptable excipient or carrier known in the art. As used herein, "pharmaceutical acceptable carriers" include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, and the like. The use of such media and agents for pharma- ceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in a therapeutic composition is contemplated. A pharmaceutical composition according to the present disclosure used to treat a subject in need thereof may also include a buffer, an agent to adjust its osmolality, and optionally one or more pharma- ceutical acceptable additives known in the art. A pharmaceutical composition according to the present disclosure used to treat a subject in need thereof, which may be conveniently provided in unit dosage form, may be prepared according to prior art techniques well known in the pharmaceutical industry, for example as detailed in the examples below. It should be understood that in addition to the ingredients specifically mentioned herein, compositions according to the present disclosure may also include other agents standard in the art having regard to the type of formulation in question.

[0135] The isolated peptide comprising the amino acid sequence set forth in SEQ ID NO:1 and / or the Bcl2 inhibitor defined herein may be administered by any route of administration known to the skilled artisan, for example intravenous (iv), or any suitable further route including intraperitoneal, subcutaneous, transdermal, topical, intramuscular, intraarticular, subconjunctival or mucosal, for example oral, intranasal or intraocular administration.

[0136] The isolated peptide or Bcl2 inhibitor as defined herein may be administered in an "effective amount" necessary to achieve the desired therapeutic result. The "effective amount" is determined by the severity of the disease, in conjunction with the therapeutic objectives, the route of administration, and the patient's general condition (age, sex, weight, and other considerations known to the attending physician). Amounts effective for this use are generally between 0.001 and 1000 mg / Kg, specifically between 0.001 mg / Kg and 0.1 mg / kg, 0.02 mg / Kg and 0.1 mg / kg, 0.03 mg / Kg and 0.1 mg / kg, 0.04 mg / Kg and 0.1 mg / kg, 0.05 mg / Kg and 0.1 mg / kg, 0.06 mg / Kg and 0.1 mg / kg, 0.07 mg / Kg and 0.1 mg / kg, 0.08 mg / Kg, 0.09 mg / Kg and 0.1 mg / kg, 0. 1mg / Kg~1mg / kg, 0.2mg / Kg~1mg / kg, 0.3mg / Kg~1mg / kg, 0.4mg / Kg~1mg / kg, 0.5mg / Kg~1mg / kg, 0.6mg / Kg~1mg / kg, 0.7mg / K g~1mg / kg, 0.8mg / Kg~1mg / kg, 0.9mg / Kg~1mg / kg, 1mg / Kg~10mg / kg, 2mg / Kg~10mg / kg, 3mg / Kg~10mg / kg, 4mg / Kg~10mg / kg, 5 mg / Kg~10mg / kg, 6mg / Kg~10mg / kg, 7mg / Kg~10mg / kg, 8mg / Kg~10mg / kg, 9mg / Kg~10mg / kg, 10mg / Kg~100mg / kg, 20mg / Kg~10 0mg / kg, 30mg / Kg~100mg / kg, 40mg / Kg~100mg / kg, 50mg / Kg~100mg / kg, 60mg / Kg~100mg / kg, 70mg / Kg~100mg / kg, 80mg / Kg~10 The range is 0 mg / kg, 90 mg / Kg to 100 mg / Kg, or 100 mg / Kg to 1000 mg / Kg, 200 mg / Kg to 1000 mg / Kg, 300 mg / Kg to 1000 mg / Kg, 400 mg / Kg to 1000 mg / Kg, 500 mg / Kg to 1000 mg / Kg, 600 mg / Kg to 1000 mg / Kg, 700 mg / Kg to 1000 mg / Kg, 800 mg / Kg to 1000 mg / Kg, or 900 mg / Kg to 1000 mg / Kg.In some specific embodiments, the amount or dose is 1 mg / Kg, 2 mg / Kg, 3 mg / Kg, 4 mg / Kg, 5 mg / Kg, 6 mg / Kg, 7 mg / Kg, 8 mg / Kg, 9 mg / Kg, 10 mg / Kg, 15 mg / kg, 20 mg / Kg, 30 mg / Kg, 40 mg / Kg, 50 mg / Kg, 60 mg / Kg, 70 mg / Kg, 80 mg / Kg, 90 mg / Kg, or 100 mg / Kg. In some specific embodiments, the amount or dose of Nerofe is 15 mg / kg. In some further embodiments, the amount of the Bcl2 inhibitor is 50 mg / kg.

[0137] In some further embodiments, an effective amount refers to the amount of Nerofe and a Bcl2 inhibitor to achieve a synergistic effect.

[0138] Single or multiple administrations on a daily, weekly or monthly schedule may be performed with the dosage levels and patterns selected by the treating physician. Additionally, administration of the compositions of the present invention may be periodic, for example, periodic administration may be performed twice a day, three times a day, or at least once a day for at least about 3 days to 3 months. The advantages of low doses will be apparent to those skilled in the art. These include, among others, a reduced risk of side effects, especially with long-term use, and a reduced risk of the patient's sensitivity to treatment decreasing. In another embodiment, treatment using the compositions of the present invention is performed after at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 14, 30, 60, 90 days of treatment, and can proceed to lifelong treatment. It should be noted that treatment may indicate the use of different doses or different periods, which will be apparent to the skilled physician.

[0139] For prophylactic applications, the compositions of the present invention may contain a prophylactically effective amount of the active ingredient. The term "prophylactically effective amount" is intended to mean an amount of a pharmaceutical composition that prevents or reduces the risk of occurrence or recurrence of a biological or medical event sought to be prevented in a tissue, system, animal or human by a researcher, veterinarian, physician or other clinician. In prophylactic applications, the compositions of the present invention are administered to a patient at risk of developing a disease state to enhance the patient's resistance. Such an amount is defined to be a "prophylactically effective dose." In this use, the exact amount again depends on the patient's health and general immunity level, but generally ranges from 0.001 to 1000 mg per administration.

[0140] The definition of the phrase "combination therapy" or "adjunctive therapy" or its use with a peptide described herein (specifically, an isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1, also referred to as "Nerofe") and at least one Bcl2 inhibitor (specifically, an inhibitor against the Bcl-2 pro-apoptotic protein) is intended to encompass sequential administration of each agent in a regimen that provides a beneficial effect of the drug combination, as well as substantially simultaneous administration (e.g., of a single formulation having a fixed ratio of these active agents or of multiple separate formulations for each agent).

[0141] Thus, in a further aspect, a fifth aspect, the present disclosure provides a method for achieving treatment, prevention, amelioration, amelioration, or delay in onset of at least one proliferative disorder in a subject treated with an inhibitor against at least one Bcl2 pro-survival protein, the method comprising administering to the subject a therapeutically effective amount of an isolated peptide comprising an amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0142] In yet another aspect, the disclosure provides a method for achieving treatment, prevention, amelioration, reduction, or delay in onset of at least one proliferative disorder in a subject treated with at least one isolated peptide comprising an amino acid sequence represented by SEQ ID NO:1, the method comprising administering to the subject a therapeutically effective amount of at least one inhibitor against Bcl2 pro-survival protein, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0143] With respect to the above aspects of the present disclosure relating to a method for effecting treatment, prevention, amelioration, reduction, or delayed onset of at least one proliferative disorder in a subject treated with at least one inhibitor against Bcl2 pro-survival protein or a method for effecting treatment, prevention, amelioration, reduction, or delayed onset of at least one proliferative disorder in a subject treated with at least one isolated peptide comprising an amino acid sequence as set forth by SEQ ID NO:1, in some embodiments, the modified isolated peptide has at least 70%, or 80%, or 90%, or 95% identity to the corresponding sequence of SEQ ID NO:1. In some embodiments, the isolated peptide consists of the amino acid sequence of SEQ ID NO:1. In some embodiments, the Bcl-2 pro-survival protein is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1, and Bcl-B / Bcl2L10. In some embodiments, the at least one inhibitor against Bcl2 pro-survival protein is at least one BH3 mimetic compound.In some embodiments, the BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-2-yl]methyl]piperazin-1-yl)benzamide (ABT-737), phenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 4-(4-{[2-(4-chlorophenyl)-5,5-dimethyl n-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene) -3-methoxy-2H-pyrrol-5-yl)-1H-indole (GX15-070), or 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852). In some embodiments, the at least one inhibitor against Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof. In some embodiments, the at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 are administered simultaneously or sequentially.In some embodiments, the method further comprises administering at least one additional anti-cancer agent. In some embodiments, the proliferative disorder is associated with Bcl2 overexpression. In some embodiments, the proliferative disorder is at least one hematological cancer or neoplasm. In some embodiments, the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0144] Thus, in a further aspect, the present disclosure provides at least one isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1 for use in a method for treating, preventing, ameliorating, alleviating, or delaying the onset of at least one proliferative disorder in a subject treated with an inhibitor against at least one Bcl2 pro-survival protein, the method comprising administering to the subject a therapeutically effective amount of an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0145] In yet another aspect, the disclosure provides a Bcl2 pro-survival protein for use in a method for achieving treatment, prevention, amelioration, reduction, or delay in onset of at least one proliferative disorder in a subject treated with at least one isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, the method comprising administering to the subject a therapeutically effective amount of at least one inhibitor of Bcl2 pro-survival protein, or any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

[0146] In some embodiments, the modified isolated peptide suitable for use in the present invention has at least 70%, or 80%, or 90%, or 95% identity to a sequence corresponding to SEQ ID NO: 1. In some embodiments, the Bcl-2 pro-survival protein suitable for use in the present invention is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1, and Bcl-B / Bcl2L10. In some embodiments, the at least one inhibitor of the Bcl2 pro-survival protein suitable for use in the present invention is at least one BH3 mimetic compound. In some embodiments, BH3 mimetic compounds suitable for use in the present invention include 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 4-(4-{[2-(4-chlorophenyl)-5, 5-Dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methyl) The compound is at least one of 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).In some embodiments, the at least one inhibitor against Bcl2 pro-survival protein suitable for use in the present invention is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof. In some embodiments, the at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide suitable for use in the present invention comprising the amino acid sequence set forth by SEQ ID NO: 1 are administered simultaneously or sequentially. In some embodiments, the use according to the present invention further comprises administering at least one additional anti-cancer agent. In some embodiments, the proliferative disorder associated with the use of the present invention is associated with Bcl2 overexpression. In some embodiments, the proliferative disorder associated with the use of the present invention is at least one hematological cancer or neoplasm. In some embodiments, the hematological cancer or neoplasm relevant to the use of the invention is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0147] As mentioned above, the present invention includes the use of different active ingredients, e.g., a peptide called "Nerofe", and at least one Bcl2 inhibitor that can be administered via different routes, dosages and combinations. More specifically, the treatment of Bcl-2 overexpression diseases and conditions with a combination of active ingredients can include separate administration of each active ingredient. Thus, the kit, offering a convenient modular format, allows the required flexibility in the above parameters.

[0148] More specifically, the kits described further herein below may include compositions as described or in separate multiple dosage unit forms as already prepared liquid topical, nasal or oral dosage forms ready for administration, or may include compositions as described as solid pharmaceutical compositions that can be reconstituted with a solvent to provide a liquid dosage form. When the kit includes a solid pharmaceutical composition that can be reconstituted with a solvent to provide a liquid dosage form (e.g., for oral administration), the kit may optionally include a reconstitution solvent. In this case, the constitutive solvent or reconstitution solvent is combined with the active ingredient to provide a liquid dosage form of each active ingredient or combination thereof. Typically, the active ingredient is soluble in the solvent to form a solution. The solvent may be, for example, water, a non-aqueous liquid, or a combination of a non-aqueous component and an aqueous component. Suitable non-aqueous components include, but are not limited to, oils, alcohols such as ethanol, glycerin, and glycols such as polyethylene glycol and propylene glycol. In some embodiments, the solvent is phosphate buffered saline (PBS).

[0149] In a further aspect, the present disclosure provides a method for producing a method for treating a cancer cell comprising: (a) at least one inhibitor against Bcl2 pro-survival protein, or any pharma- ceutically acceptable salt thereof, which may be in a first dosage form; and (b) an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, which may be in a second dosage form; A kit is provided, comprising:

[0150] In some embodiments, a modified isolated peptide suitable for use in a kit of the present disclosure or invention has at least 70%, or 80%, or 90%, or 95% identity to a sequence corresponding to SEQ ID NO:1.

[0151] In some specific embodiments, the isolated peptide consists of the amino acid sequence of SEQ ID NO:1.

[0152] In some further embodiments, the Bcl-2 pro-survival protein associated with the kit of the invention is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1, and Bcl-B / Bcl2L10.

[0153] In some more particular embodiments, the at least one inhibitor against Bcl2 pro-survival protein is at least one BH3 mimetic compound.

[0154] In some more specific embodiments, the BH3 mimetic compounds suitable for the kits of the present invention are 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 4-(4-{[2-(4-chlorophenyl) -5,5-dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methyl) The compound is at least one of 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).

[0155] In some particular embodiments, the at least one inhibitor against Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

[0156] In some further embodiments, the kits according to the present disclosure are for use in a method for treating, preventing, ameliorating, reducing, or delaying the onset of at least one proliferative disorder.

[0157] In some embodiments, at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1 are in separate unit dosage forms in a kit for use according to the invention.

[0158] In some further embodiments, separate unit dosage forms of at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 are administered simultaneously or sequentially.

[0159] In some other embodiments, the proliferative disorder associated with the kit for use of the invention is associated with Bcl2 overexpression.

[0160] In some further embodiments, the proliferative disorder is at least one hematological cancer or neoplasm.

[0161] In some specific embodiments, the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumors, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

[0162] The term "treatment or prevention" refers to the full range of therapeutically positive effects of administration to a subject, including inhibition, reduction, alleviation, and mitigation of proliferative disorder symptoms or undesirable side effects of such proliferative disorder-associated disorders. More specifically, treatment or prevention includes preventing or postponing the onset of disease, preventing or postponing the onset of symptoms, and / or reducing the severity of such symptoms that develop or are expected to develop. These further include ameliorating existing symptoms, preventing further symptoms, and ameliorating or preventing the underlying metabolic causes of symptoms.

[0163] As used herein, "disease," "disorder," "condition," and the like, when they relate to the health of a subject, are used interchangeably and have the meanings ascribed to each such term.

[0164] The present invention relates to the treatment of subjects or patients in need of treatment. By "patient" or "subject in need" is meant any organism that may be affected by the above-mentioned conditions and for which the treatment methods described herein are desired, including humans, domestic and non-domestic mammals, such as canine and feline subjects, bovine, simian, equine and murine elephants, rodents, domestic birds, aquaculture, fish and exotic ornamental fish. It should be understood that the subject to be treated can also be any reptile or zoo animal. More specifically, the methods and compositions of the present invention are intended to be directed to mammals. By "mammalian subject" is meant any mammal for which the proposed treatment is desired, including human, equine, canine and feline subjects, most specifically humans. It should be noted that, particularly in the case of non-human subjects, the methods of the present disclosure may be carried out using administration directly to the digestive tract of the subject in need thereof, via injection, drinking water, feed, nebulization, oral gavage, etc. It should be further noted that, particularly in the case of human subjects, administration of the compositions of the present invention to the patient includes both self-administration and administration to the patient by another person.

[0165] All definitions defined and used herein are understood to govern any dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0166] As used herein, the term "about" refers to values ​​that may deviate up to 1%, more specifically 5%, more specifically 10%, more specifically 15%, and in some cases up to 20% higher or lower than the stated value, the deviation range including integer values, where applicable, non-integer values ​​also constitute a continuous range. As used herein, the term "about" refers to ±10%.

[0167] The indefinite articles "a" and "an," as used in this specification and the claims, should be understood to mean "at least one," unless clearly indicated otherwise. It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0168] The phrase "and / or" as used in the specification and claims should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctive in some cases and disjunctive in other cases. Multiple elements listed with "and / or" should be construed in the same manner, i.e., "one or more" of the elements so conjoined. Other elements other than the elements specifically identified by the "and / or" clause may optionally be present, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to "A and / or B," when used in conjunction with open-ended language such as "comprising," may refer in one embodiment to only A (optionally including elements other than B), in another embodiment to only B (optionally including elements other than A), in yet another embodiment to both A and B (optionally including other elements), etc.

[0169] As used herein and in the claims, "or" should be understood to have the same meaning as "and / or" defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as being inclusive, i.e., including at least one of, but also including more than one, and optionally including additional unlisted items, of a plurality of elements or a list of elements. Only terms clearly indicated to the contrary, such as "only one of" or "exactly one of," or, when used in the claims, "consisting of," refer to the inclusion of exactly one element of a plurality of elements or a list of elements. In general, the term "or" as used herein, "either," "one of," "only one of," or "consisting essentially of," when preceded by a term of exclusivity, shall be interpreted only as indicating exclusive alternatives (i.e., "one or the other, but not both") and, when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0170] As used herein and in the claims, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not necessarily excluding any combination of elements in the list of elements. This definition also allows for elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified, may optionally be present. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B" or, equivalently, "at least one of A and / or B") can refer in one embodiment to at least one, optionally more than one, A, and no B (optionally including elements other than B); in another embodiment to at least one, optionally more than one, B, and no A (optionally including elements other than A); in yet another embodiment to at least one, optionally more than one, A, and at least one, optionally more than one, B (and optionally including other elements); and so forth.

[0171] Also, unless expressly indicated to the contrary, it is to be understood that in any method claimed herein that includes two or more steps or actions, the order of the method steps or actions is not necessarily limited to the order in which the method steps or actions are recited.

[0172] Throughout this specification and the following examples and claims, all transitional phrases such as "comprising," "including," "carrying," "having," "containing," "involving," "holding," "composed of," and the like, are to be understood to be open-ended, i.e., to mean including but not limited to. Specifically, it is to be understood to mean the inclusion of a stated integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps. Only the transitional phrases "consisting of" and "consisting essentially of" are intended to be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures. More specifically, the terms "comprises," "comprising," "includes," "including," "having," and conjugations thereof, mean "including but not limited to." The term "consisting of" means "including and limited to." The term "consisting essentially of" means that a composition, method, or structure may include additional components, steps, and / or moieties, but only if the additional components, steps, and / or moieties do not materially alter the basic and novel characteristics of the claimed composition, method, or structure.

[0173] It should be noted that various embodiments of the present invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the present invention. Thus, the description of a range should be considered to have all possible subranges specifically disclosed as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to specifically disclose subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range. Whenever a numerical range is given herein, it is meant to include any cited numbers (fractional or integer) within the given range. The phrases "ranging / ranges between" a first indicated number and a second indicated number and "ranging / ranges from" a first indicated number to a second indicated number are used interchangeably herein and are meant to include the first and second indicated numbers and all fractions and integers therebetween.

[0174] As used herein, the term "method" refers to manners, means, techniques and procedures for accomplishing a given task, including, but not limited to, manners, means, techniques and procedures that are either known to a practitioner in the chemical, pharmacological, biological, biochemical and medical arts or that can be readily developed from known manners, means, techniques and procedures.

[0175] It is understood that certain features of the invention that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described for brevity in the context of a single embodiment may also be provided separately or in any suitable subcombination, or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperable without those elements.

[0176] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below are supported experimentally in the following examples.

[0177] Although disclosed and described, it should be understood that the invention is not limited to the specific examples, method steps, and compositions disclosed herein, as such method steps and compositions may vary somewhat. It should also be understood that the terminology used herein is used only for the purpose of describing particular embodiments, and is not intended to be limiting, since the scope of the invention is limited only by the appended claims and equivalents thereof.

[0178] The following examples are representative of techniques employed by the inventors in carrying out aspects of the present invention. It should be understood that while these techniques are illustrative of preferred embodiments for carrying out the invention, those skilled in the art will recognize that many modifications may be made upon consideration of this disclosure without departing from the spirit and intended scope of the invention. EXAMPLES

[0179] Experimental procedure * Preparation of dTCApF peptide and composition containing same The peptide dTCApF or NEROFE™ (both terms are used interchangeably herein and refer to the same peptide as above) is a 14 amino acid residue long peptide in which all of the amino acid residues are in the D-configuration and has the amino acid sequence Trp Trp Thr Phe Phe Leu Pro Ser Thr Leu Trp Glu Arg Lys (or in the one letter code WWTFFLPSTLWERK, as represented by SEQ ID NO:1).

[0180] The peptide was synthesized as follows: The final drug product, dTCApF acetate, for use in clinical trials was manufactured, packaged, tested, labeled, and released under good manufacturing practice (GMP) by Nextar Ltd., Israel.

[0181] * Western blot of cleaved caspase 3 in U937 cells treated with Nerofe and ABT-737 material: For cell proliferation: -Growth medium- · RPMI Media 1640(Gibco 21875-034) 50mL heat inactivated FBS (Biological Industries, catalog number 04-121-1A). 0.5mL of Amphotericin B 2 500pg / mL (Biological industries, catalog number 03-029-1). Penicillin-streptomycin solution (Biological Industries, catalog number 03-031)

[0182] -Treatment medium- Growth medium - 10% Mannitol (Sigma Catalog No. M4125-500G)

[0183] After adding mannitol, the medium is filtered through a 0.2 pm filter. -175cm 2 Culture flask -Nerofe 20 mg / mL in mannitol -ABT-737 (Cayman 11501) -U937 cell line

[0184] For Western Blot - Lysis buffer containing: · Ripa buffer (Sigma R0278) · Protease inhibitor cocktail (Sigma P2714). Phosphatase inhibitor (Sigma P5726) Benzonase® Nuclease, >90% pure (70746 Millipore), 25U / mL -4x LDS sample buffer (Invitrogen, Catalog No. NP0007) -Sample Reducing Agent (Invitrogen Catalog No. NP0009, 10x concentrated) -4-12% Bis-Tris gel (Millipore, Cat. No. MMMP41G10) -10x MES transfer buffer (Millipore MMMPTRB) -PVDF membrane (Immobilon-P sq 0.2um) -TBST buffer: TBS (J640 Amersco) + 0.05 Tween-20 -5% skim milk in TBST -Primary antibody diluted 1:500 in 5% skim milk in TBST: Anti-cleaved caspase 3 antibody (ab32042 abcam) -Secondary antibody diluted 1:10,000 in 5% skim milk in TBST: · Anti-rabbit IgG, HRP binding antibody (Cells signaling 7074) -SuperSignal(TM)West Femto Maximum Sensitivity Substrate(Thermo Scientific 34095)

[0185] Cell culture - passaging: Cells were grown at 37 °C, 5% CO2, and 100% humidity until they reached a density of 70-80%. Cells were resuspended in fresh medium and then grown for 2-3 days until they reached a density of 70-80%. The passaging procedure was repeated.

[0186] Cell treatment: Cells were seeded at 700K cells / 75CM2 flask in 13 ml of growth medium in four different flasks and incubated overnight. Mannitol was added to a final mannitol concentration of 5% mannitol. Cells were then treated as follows: 1- Control (untreated) 2-Nerofe 50μg / mL 3- No processing (ABT-737 the next day) 4-Nerofe 50μg / mL (ABT-737 the next day)

[0187] The cells were then incubated for 24 hours and treated as follows. 1- Control (untreated) 2- No treatment (treated with Nerofe the day before) 3-ABT-737 1μM 4-ABT-737 1μM (treated with Nerofe the day before)

[0188] The cells were then incubated for 24 hours and harvested for Western blot as previously described (Mahmood, T., & Yang, PC (2012). Western blot: technique, theory, and trouble shooting. North American journal of medical sciences sciences, 4(9), 429-434).

[0189] * Cell viability assay of U937 cells treated with Nerofe and Azacitidine (Resazurin) material: -Cell culture medium: 10% FBS (Biological Industries, Catalog No. 04-121-1A) DMEM (Gibco, catalog number 41965) 1mM Sodium Pyruvate (Biological Industries) 100 μg / mL penicillin and 100 μg / mL streptomycin (Biological Industries) 250ng / mL Amphotericin B (Biological Industries) -Treatment medium- Culture medium: 10% Mannitol (Sigma, Cat. No. M4125-500G)

[0190] After the addition of mannitol, the medium is filtered through a 0.2 μm filter. -Cytotoxicity assay kit (colorimetric): abl 12118 abeam · Cell line: U937 · 2mg / mL Nerofe stock solution: Dilute 100μl of 20mg / mL Nerofe in 900μl of cell treatment medium. 5-Azacytidine (Sigma A2385, freshly diluted to 50mg / ml in treatment medium)

[0191] [Table 1]

[0192] Cells were seeded at 20000 cells / well in 100 μl growth medium in black 96-well plates with clear bottom and incubated for 24 hours. Cells were treated with Nerofe and Azacitidine in 100 μl cell treatment medium with 10% mannitol in a final volume of 200 μl and a final mannitol concentration of 5%. Cells were incubated for 48 hours. A volume of 20 μl of Cytotoxicity Assay Kit (Colorimetric) ab 12118 was added to each well and cells were incubated for 4 hours. Plates were read using a Biotek Synergy-HT plate reader at excitation 530, emission 590, sensitivity 40. Percentage of cell viability was calculated compared to control.

[0193] * Cell viability assay (resazurin) of U937 cells treated with Nerofe and ABT-737 material The materials were the same as above, except that ABT-737 (Cayman 11501) was substituted for 5-azacytidine.

[0194] [Table 2]

[0195] Cells were seeded at 20000 cells / well in 100 μl growth medium in black 96-well plates with clear bottom and incubated for 24 hours. Cells were treated with Nerofe in 100 μl cell treatment medium with 10% mannitol in a final volume of 200 μl and a final mannitol concentration of 5%. Cells were incubated for 24 hours. Cells were treated with ABT-737 in a final volume of 225 μl by adding 25 μl treatment medium with 5% mannitol without FBS to each well. Cells were incubated for 24 hours. A volume of 25 μl of Cytotoxicity Assay Kit (Colorimetric) ab 12118 was added to each well and cells were incubated for 4 hours. Plates were read using a Biotek Synergy-HT plate reader at excitation 530, emission 590, sensitivity 40. Percentage of cell viability was calculated compared to control.

[0196] * Western blot of cleaved caspase 3 in CT26 cells treated with Nerofe and ABT-737 material: For cell proliferation: -Growth medium- · RPMI Media 1640(Gibco 21875-034) 50mL heat inactivated FBS (Biological Industries, catalog number 04-121-1A). 0.5mL of Amphotericin B, 2500pg / mL (Biological industries, catalog number 03-029-1). Penicillin-streptomycin solution (Biological Industries, catalog number 03-031) -Treatment medium- Growth medium - 5% Mannitol (Sigma, Cat. No. M4125-500G)

[0197] After adding mannitol, the medium is filtered through a 0.2 μm filter. -Trypsin EDTA (Biological industries, catalog number 03-052-1B). -175cm 2 Culture flask -Nerofe 20mg / mL in mannitol (use aliquots, avoid repeated freeze-thaw cycles). -ABT-737 (Cayman 11501) -CT26 cell line

[0198] For Western Blot - Lysis buffer containing: · Ripa buffer (Sigma R0278) · Protease inhibitor cocktail (Sigma P2714). Phosphatase inhibitor (Sigma P5726) Benzonase® Nuclease, >90% pure (70746 Millipore), 25U / mL -4x LDS sample buffer (Invitrogen, Catalog No. NP0007) -Sample Reducing Agent (Invitrogen, Catalog No. NP0009, 10x concentrated) -4-12% Bis-Tris gel (Millipore, Cat. No. MMMP41G10) -10x MES transfer buffer (Millipore MMMPTRB) -PVDF membrane (Immobilon-P sq 0.2μm) -TBST buffer: TBS (J640 Amersco) + 0.05% Tween-20 -5% skim milk in TBST -Primary antibody diluted 1:1000 in 5% skim milk in TBST: · Anti-cleaved caspase 3 antibody (asp 175) Cell Signaling #9661 -Secondary antibody diluted 1:10,000 in 5% skim milk in TBST: · Anti-rabbit IgG, HRP binding antibody (Cells signaling 7074) -SuperSignal(TM) West Femto Maximum Sensitivity Substrate(Thermo Scientific34095)

[0199] Cell culture - passaging: Cells were grown at 37 °C, 5% CO2, and 100% humidity until they reached a density of 70-80%. Cells were treated with trypsin. Cells were resuspended in fresh medium and then grown for 2-3 days until they reached a density of 70-80%. The passaging procedure was repeated.

[0200] Cell treatment: Cells were seeded at 1M cells / 75 CM2 flasks in 25 mL of growth medium in four different flasks and incubated overnight. The medium was changed. Cells were then treated as follows: 1- Control (untreated) 2-Nerofe 50μg / mL 3-No treatment (for ABT) 4-Nerofe 50μg / mL (for ABT) 5-No treatment (for ABT) 6-Nerofe 50μg / mL (for ABT)

[0201] The cells were then incubated for 24 hours and treated as follows. 1- Control (untreated) 2-No processing 3-No processing 4-No processing 5-ABT 1μM 6-ABT 1μM

[0202] The cells were then incubated for 24 hours and treated as follows. 1- Control (untreated) 2-Nerofe 50μg / mL 3-ABT 1μM 4-ABT 1μM+Nerofe 50μg / mL 5-No processing 6-Nerofe 50μg / mL

[0203] Cells were then harvested for Western blotting as previously reported (Mahmood, T., & Yang, PC (2012). Western blot: technique, theory, and trouble shooting. North American journal of medical sciences sciences, 4(9), 429-434).

[0204] * Cell viability assay (resazurin) of CT26 cells treated with Nerofe and ABT-737 material -Cell culture medium: 10% FBS (Biological Industries, Catalog No. 04-121-1A) DMEM (Gibco, catalog number 41965) 1mM Sodium Pyruvate (Biological Industries) 100 μg / mL penicillin and 100 μg / mL streptomycin (Biological Industries) 250ng / mL Amphotericin B (Biological Industries) -Treatment medium- 5% mannitol (Sigma, Cat. No. M4125-500G) in culture medium containing FBS or 5% Mannitol in culture medium without FBS (Sigma, Cat. No. M4125-500G)

[0205] After the addition of mannitol, the medium is filtered through a 0.2 μm filter. -Cytotoxicity assay kit (colorimetric): abl 12118 abeam · Cell line: CT26 · 2mg / mL Nerofe stock solution: Dilute 100μl of 20mg / mL Nerofe in 900μl of cell treatment medium. ABT-737 (Cayman 11501)

[0206] [Table 3]

[0207] Cells were seeded at 5,000 cells / well in black 96-well plates with clear bottom in 100 μl of growth medium. Cells were incubated for 24 hours. Medium was removed and cells were treated with Nerofe in 100 μl of cell treatment medium with FBS. Cells were incubated for 24 hours. Cells were then treated with ABT-737 in 48-hour treatment wells in a final volume of 125 μl by adding 25 μl of treatment medium with 5% mannitol without FBS to each well. Cells were incubated for 24 hours. Cells were treated with ABT-737 and Nerofe in 24-hour treatment wells in a final volume of 150 μl by adding 25 μl of treatment medium with 5% mannitol without FBS to each well. Cells were incubated for 24 hours.

[0208] A volume of 25 μl of Cytotoxicity Assay Kit (Colorimetric) ab 12118 was added to each well and the cells were incubated for 4 hours. The plates were read using a Biotek Synergy-HT plate reader at excitation 530, emission 590, and sensitivity 40. The percentage of cell viability was calculated compared to the control.

[0209] Example 1 Synergistic effect of Nerofe and BCL2 inhibitors in inducing apoptosis in U937 cells As mentioned above, a peptide called "T101" encoded by a cDNA specific to the human thymus has been identified and reported to, among other things, reduce cancer tumor size. Furthermore, a peptide derivative of T101, referred to herein as dTCAPF (or "Nerofe"), has been reported to be involved in cancer cell migration [8] and induction of ER stress, which contributes to promoting cell death [9].

[0210] Specifically, Nerofe is a peptide having an amino acid sequence of Trp Trp Thr Phe Phe Leu Pro Ser Thr Leu Trp Glu Arg Lys, all of which are D-form, as shown in SEQ ID NO: 1, and was prepared as described above.

[0211] The effect of Nerofe and ABT-737, either alone or in combination, on the activation of caspase 3 in U937 cells was examined. As shown in Figure 1, when Nerofe was combined with a BCL2 inhibitor, i.e., ABT-737, a strong synergistic effect was observed on the activation of caspases, a marker of apoptosis. Without wishing to be bound by theory, it is believed that the BCL2 protein is the primary inhibitor of the apoptotic effect of Nerofe.

[0212] Example 2 Synergistic effect of Nerofe and BCL2 inhibitors in inducing cell death in U937 cells The effect of Nerofe and BCL2 inhibitor, i.e., ABT-737, either alone or in combination, on cell viability of U937 cells was investigated at different concentrations, i.e., 0.5, 1, or 2 μM of ABT-737. As shown in Figure 2, a strong synergistic effect on induction of cell death was observed when Nerofe was combined with ABT-737.

[0213] Example 3 No synergy between Nerofe and azacytidine in inducing cell death was observed The effect of Nerofe and Azacitidine, either alone or in combination, on the cell viability of U937 cells was examined at different concentrations, i.e., 50 or 100 nM Azacitidine. As shown in Figure 3, when Nerofe was combined with Azacitidine, no synergistic effect was observed on the induction of cell death. Without wishing to be bound by theory, the antineoplastic activity of Nerofe does not appear to be due to the same mechanism of action as Azacitidine. Azacitidine is a chemical analogue of the nucleoside cytidine, which is present in DNA and RNA and has antineoplastic activity through two mechanisms: at low doses, by inhibiting DNA methyltransferase, causing DNA hypomethylation, and at high doses, by direct cytotoxicity due to incorporation into DNA and RNA, resulting in cell death. On the other hand, Nerofe is believed to act through the production of free oxygen radicals, which disrupt the Golgi apparatus and induce unrepaired ER stress in cancer cells overexpressing T1 / ST2 receptors.

[0214] Example 4 Synergistic effect of Nerofe and BCL2 inhibitors in inducing cell death in CT26 cells The effect of Nerofe and BCL2 inhibitor, i.e., ABT-737, either alone or in combination, on cell viability of CT26 cells was examined at a concentration of 1 μM ABT-737. As shown in Figure 4, a strong synergistic effect on the induction of cell death was observed when Nerofe was combined with ABT-737.

[0215] Example 5 Synergistic effect of Nerofe and BCL2 inhibitors in inducing apoptosis in CT26 cells The effects of Nerofe and ABT-737, either alone or in combination, on the activation of cleaved caspase-3 in CT26 cells were examined by Western blot analysis. As shown in Figure 5, when Nerofe was combined with a BCL2 inhibitor, i.e., ABT-737, a strong synergistic effect on caspase activation was observed.

[0216] Example 6 Effect of Nerofe with additional Bcl2 inhibitor The effect of Nerofe with an additional Bcl2 inhibitor, such as ABT-199 (venetoclax), ABT-263 (navitoclax), GX15-070 (obatoclax) or A-1331852, in inducing apoptosis and / or cell death is further tested, for example as described above.

[0217] Example 7 Effect of combining Nerofe with a BCL2 inhibitor in a mouse model Thirty-two Blab / C females were inoculated subcutaneously (SC) with 100,000 CT26 cells. Tumors were 50 mm 3 Once the mice reach a volume of 1000 mg / kg, they are divided into the following groups: - 8 mice - control - 8 mice - Nerofe (15 mg / kg, intraperitoneally (IP), 3 times a week) - 8 mice - ABT-737 (50mg / Kg) IP, 3 times per week - 8 mice - Nerofe (15mg / kg, IP, 3 times per week + ABT-737 (50mg / Kg) IP, 3 times per week Mice are weighed twice weekly and tumor volumes are measured twice weekly.

[0218] Example 8 Effect of combining Nerofe with an additional BCL2 inhibitor in a mouse model Thirty-two Blab / C females were inoculated subcutaneously (SC) with 100,000 CT26 cells. Tumors were 50 mm 3 Once the mice reach a volume of 1000 mg / kg, they are divided into the following groups: - 8 mice - control - 8 mice - Nerofe (15 mg / kg, intraperitoneally (IP), 3 times a week) - 8 mice - ABT-199 (venetoclax) IP, 3 times weekly - 8 mice - ABT-263 (Navitoclax) IP, 3x per week; IP, 3x per week - 8 mice - GX15-070 (Ovatoclax) IP, 3x per week; IP, 3x per week - 8 mice - A-1331852, IP, 3x per week; IP, 3x per week - 8 mice - Nerofe, IP, 3x per week + ABT-199 (venetoclax), IP, 3x per week - 8 mice - Nerofe, IP, 3 times per week + ABT-263 (Navitoclax), IP, 3 times per week - 8 mice - Nerofe, IP, 3x per week + GX15-070 (Obatoclax), IP, 3x per week - 8 mice - Nerofe, IP, 3 times per week + A-1331852 (Obatoclax), IP, 3 times per week Mice are weighed twice weekly and tumor volumes are measured twice weekly.

Claims

1. A combination composition comprising at least one inhibitor against B-cell lymphoma 2 (Bcl2) pro-survival protein, an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier, diluent, excipient and / or additive.

2. The composition of claim 1 , wherein the modified isolated peptide has at least 70%, or 80%, or 90%, or 95% identity to the corresponding sequence of SEQ ID NO:

1.

3. 3. The composition of claim 1 or 2, wherein the Bcl-2 pro-survival protein is at least one of Bcl-2, B-cell lymphoma extra-large (Bcl-xL), myeloid cell leukemia-1 (Mcl-1), B-cell lymphoma-w (Bcl-w), B-cell lymphoma 2-associated protein A1 (Bcl2A1), and B-cell lymphoma 2-like 10 (Bcl-B / Bcl2L10).

4. The composition of any one of claims 1 to 3, wherein the at least one inhibitor against Bcl2 pro-survival protein is at least one Bcl-2 homology 3 (BH3) mimetic compound.

5. The BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohex-1-en-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)methyl)piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 5. The composition of claim 4, which is at least one of 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852), or a combination thereof.

6. 6. The composition of claim 5, wherein the at least one inhibitor of Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), or any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

7. 7. The composition of any one of claims 1 to 6 for use in a method for treating, preventing, ameliorating, alleviating or delaying the onset of at least one proliferative disorder.

8. The composition of claim 7 for use, wherein the proliferative disorder is associated with Bcl2 overexpression.

9. 9. The composition of claim 7 or 8 for use, wherein the proliferative disorder is at least one blood cancer or neoplasm.

10. 10. The composition of claim 9, wherein the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumor, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

11. A method for achieving treatment, prevention, amelioration, alleviation, or delay in the onset of at least one proliferative disorder in a subject in need thereof, comprising administering a therapeutically effective amount of at least one inhibitor of Bcl2 pro-survival protein and an isolated peptide comprising an amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharmaceutically acceptable salt thereof, or any composition, kit, or combination thereof.

12. The method of claim 11, wherein the modified isolated peptide has at least 70%, or 80%, or 90%, or 95% identity to the corresponding sequence of SEQ ID NO:

1.

13. 13. The method of claim 11 or 12, wherein the Bcl-2 pro-survival protein is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1 and Bcl-B / Bcl2L10.

14. The method according to any one of claims 11 to 13, wherein said at least one inhibitor against Bcl2 pro-survival protein is at least one BH3 mimetic compound.

15. The BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-4,4-di 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohex-1-enyl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-2-yl)methyl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-2-yl)methyl)- 15. The method of claim 14, wherein the compound is at least one of 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).

16. 16. The method of claim 15, wherein the at least one inhibitor of Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), or any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

17. 17. The method according to any one of claims 11 to 16, wherein said at least one inhibitor against Bcl2 pro-survival protein and the isolated peptide comprising the amino acid sequence as set forth by SEQ ID NO:1 are administered simultaneously or sequentially.

18. The method of any one of claims 11 to 17, further comprising administering at least one additional anti-cancer agent.

19. The method of any one of claims 11 to 18, wherein the proliferative disorder is associated with Bcl2 overexpression.

20. The method of any one of claims 11 to 19, wherein the proliferative disorder is at least one hematological cancer or neoplasm.

21. 21. The method of claim 20, wherein the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumor, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

22. At least one inhibitor of Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, for use in a method for treating, preventing, ameliorating, alleviating, or delaying the onset of at least one proliferative disorder.

23. (a) at least one inhibitor against Bcl2 pro-survival protein, or any pharma- ceutically acceptable salt thereof, optionally in a first dosage form; (b) an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, which may be in a second dosage form; Including the kit.

24. 24. The kit of claim 23, wherein the modified isolated peptide has at least 70%, or 80%, or 90%, or 95% identity to the corresponding sequence of SEQ ID NO:

1.

25. 25. The kit of claim 23 or 24, wherein the Bcl-2 pro-survival protein is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bcl2A1 and Bcl-B / Bcl2L10.

26. The kit according to any one of claims 23 to 25, wherein said at least one inhibitor against Bcl2 pro-survival protein is at least one BH3 mimetic compound.

27. The BH3 mimetic compound is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)benzamide (ABT-737), 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl 4-(4-{[2-(4-chlorophenyl)-5,5-dimethylcyclohex-1-enyl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)-amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]-pyridin-5-yloxy)benzamide (ABT-199), 2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-4-yl)methyl)piperazin-1-yl)-N-(4-{[(2R)-4-(morpholin-4-yl)-1-(phenylsulfanyl)butan-2-yl]amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl)benzamide (ABT-263), ...

27. The kit of claim 26, wherein the compound is at least one of 3-[1-(1-adamantylmethyl)-5-methylpyrazol-4-yl]-6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]pyridine-2-carboxylic acid (A-1331852).

28. 28. The kit of claim 27, wherein the at least one inhibitor of Bcl2 pro-survival protein is 4-{4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}-N-(4-{[(2R)-4-(dimethylamino)-1-(phenylsulfanyl)butan-2-yl]amino}-3-nitrobenzene-1-sulfonyl)-benzamide (ABT-737), or any pharma- ceutically acceptable salt or hydrate thereof, or any stereoisomer or salt thereof.

29. 29. The kit of any one of claims 23 to 28 for use in a method for treating, preventing, ameliorating, reducing or delaying the onset of at least one proliferative disorder.

30. 30. The kit of claim 29 for use, wherein said at least one inhibitor against Bcl2 pro-survival protein and said isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1 are in separate unit dosage forms.

31. The kit of claim 30 for use, wherein the separate unit dosage forms of at least one inhibitor against Bcl2 pro-survival protein and an isolated peptide comprising the amino acid sequence set forth by SEQ ID NO:1 are administered simultaneously or sequentially.

32. The kit according to any one of claims 29 to 31 for use, wherein the proliferative disorder is associated with Bcl2 overexpression.

33. 33. The kit of any one of claims 29 to 32 for use, wherein the proliferative disorder is at least one hematological cancer or neoplasm.

34. 34. The kit of claim 33 for use, wherein the hematological cancer or neoplasm is at least one of chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), mutated KRAS tumor, lymphoma, multiple myeloma, blastic plasmacytoid dendritic cell neoplasm, T-cell prolymphocytic leukemia, or myelodysplasia.

35. A method for achieving treatment, prevention, amelioration, mitigation, or delay in onset of at least one proliferative disorder in a subject treated with at least one inhibitor against Bcl2 pro-survival protein, comprising administering to the subject a therapeutically effective amount of an isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

36. A method for achieving treatment, prevention, amelioration, reduction, or delay in onset of at least one proliferative disorder in a subject treated with at least one isolated peptide comprising the amino acid sequence represented by SEQ ID NO:1, comprising the step of administering to the subject a therapeutically effective amount of at least one inhibitor against Bcl2 pro-survival protein, any functional derivative thereof, or a pharma- ceutically acceptable salt thereof, or any composition, kit, or combination thereof.

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