Pharmaceutical combination product comprising kinase inhibitor and autophagy inducer and uses thereof

Abstract

The present invention relates to pharmaceutical products and in particular to pharmaceutical combination products. The invention further relates to uses of such products in medical treatment. Embodiments of the invention have been particularly developed as pharmaceutical combination products of an inhibitor of a tyrosine kinase and an autophagy inducer, preferably for use in the treatment of cancer and will be described hereinafter with reference to this application. However, it will be appreciated that the invention is not limited to this particular field of use.

Description

Pharmaceutical combination product comprising kinase inhibitor and autophagy inducer and uses thereofFIELD OF THE INVENTIONThe present invention relates to pharmaceutical products and in particular to pharmaceutical combination products. The invention further relates to uses of such products in medical treatment. Embodiments of the invention have been particularly developed as pharmaceutical combination products of an inhibitor of a tyrosine kinase and an autophagy inducer, preferably for use in the treatment of cancer and will be described hereinafter with reference to this application. However, it will be appreciated that the invention is not limited to this particular field of use.BACKGROUND OF THE INVENTIONAny discussion of the background art throughout the specification should in no way be considered as an admission that such art is widely known or forms part of common general knowledge in the field.BRCA-1 associated protein 1 (BAP1) is a tumour suppressor gene, germline mutations of which confer a predisposition to various malignancies. BAP1 encodes for a multi-domain protein, BAP1 , which contains an N-terminal ubiquitin carboxyl hydrolase (UCH) domain, a host cell factor 1 (HCF-1 ) binding motif (HBM) domain, and a variety of other binding regions for various protein interactions. BAP1 functions as a tumour suppressor deubiquitinating enzyme, modulating gene transcription, cellular differentiation, and DNA damage repair, apoptosis and cell metabolism.The occurrence of two rare and otherwise unrelated malignancies, uveal melanoma (UM) and malignant pleural mesothelioma (MPM) within families led to the identification of a cancer predisposition syndrome implicating germline mutations in BAP1, now known as BAP1 tumor predisposition syndrome (BAP1 -TPDS). Germline BAP1 inactivation is classified an autosomal dominant disease with high penetrance and increased susceptibility to MPM, cutaneous melanoma, UM, clear-cell renal cell carcinoma (ccRCC), cholangiocarcinoma, basal cell carcinoma, and likely other malignancies.While there has been increased interest in the mechanisms by which BAP1 inactivation leads to carcinogenesis, and while screening guidelines have been proposed for patients harboring germ line BAP1 mutations to aid with early cancer detection, no specific treatments for cancers in which BAP1 function is lost (be it due to inactive or no protein being produced) are currently available. This is particularly worrisome as inactivation of BAP1 leads to more aggressive tumors, tumor metastasis and poor patient survival.Uveal melanoma (UM) is the most common primary intraocular malignancy in adults with an estimated annual incidence in Central Europe of 0.8 per 100,000 inhabitants. While primary tumors are usually treated successfully by surgical resection or radiotherapy, around 50% of patients develop metastatic disease within few years after diagnosis. Once metastasized, median survival is very low, ranging between 3.7 to 7 months, and has not changed during the past decades due to a persisting lack of effective treatment options. UM tumors are characterized by mutually exclusive mutations in two homologous G-protein alpha subunits involved in the MAPK / ERK pathway, GNAQ and GNA11 , which are considered to be early events and key drivers of UM development. One of the most important prognostic factors is monosomy of chromosome 3 (M3), which is strongly correlated with metastatic risk. UM tumors can be classified in those with mutually exclusive mutations in EIF1AX or SF3B1 (and associated with better prognosis) and in tumors with mutations in the tumor suppressor gene BAP1. In contrast to metastatic cutaneous melanoma, metastatic UM shows limited response to immune checkpoint blockade, mainly because of the special immune system and the different connection to the vascular system in the eye. Despite clinical trials using conventional chemotherapy, immunomodulatory, or targeted therapies, no reliable treatment is currently available for metastatic UM.Renal cell carcinomas are characterized by mutations in epigenetic genes involved in chromatin remodeling. Kidney cancer is among the ten most prevalent cancers arising in Western countries, with clear-cell renal cell carcinoma (ccRCC) being the most frequent subtype (75%). About one third of ccRCC patients present with metastatic disease at diagnosis, and another 30% will develop metastases after surgery. When metastatic, ccRCC remains largely incurable. An early event in ccRCC development is the inactivation of the von Hippel-Lindau (pVHL) pathway, although several studies implicated mutations of epigenetic modifiers, such as SETD2 and PBRM1. It isnoteworthy that BAP1 is inactivated in 15% of ccRCCs. Interestingly, the four tumor suppressor genes VHL, SETD2, PBRM1 and BAP1 are closely located on chromosome 3p, an area deleted in more than 90% of ccRCC patients. Most importantly, mutations in BAP1 are mutually exclusive with mutations in PBRM1 and are also associated with different prognosis: Loss of BAP1 in ccRCC is associated with high tumor grade, activation of mTORCI and poor patient survival, whereas tumors with loss of PBRM1 are associated with low grade and better patient survival (Pe a-Llopis et al. Nat. Genet. 2012).It has been established that the tumor suppressor gene BAP1 is mutated in about 80% of metastatic UMs, in about 60% of malignant pleural mesotheliomas, in about 25% of intrahepatic cholangiocarcinomas, in about 15% of ccRCCs but also in other types of cancer at lower frequencies. However, BAP1 remains the cancer gene whose mutation and inactivation is closely associated with low patient survival and has been directly implicated for promote metastasis. Thus, tumors with BAP1 loss, be it due to inactive or no protein being produced, represent a group of tumors characterized by high aggressiveness and poor prognosis.Accordingly, there is a need in the art for improved pharmaceutical products for the treatment of cancer, such as the treatment of malignant pleural mesothelioma (MPM), cutaneous melanoma, uveal melanoma (UM), clear-cell renal cell carcinoma (ccRCC), cholangiocarcinoma, basal cell carcinoma, and likely other malignancies, in particular of cancers with BAP1 loss.It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative. In particular, it is an object of the present invention to provide improved pharmaceutical products for the treatment of cancer.SUMMARY OF THE INVENTIONThe present invention provides a solution to the above-formulated problem based on the inventors’ surprising finding that pharmaceutical combination products comprising: (a) an inhibitor of tyrosine kinase SRC, which is often deregulated in thecontext of cancer; and (b) an inducer of autophagy, are effective in the treatment of cancer (such as, e.g., in cancers with BAP1 loss).Accordingly, in a first aspect, the present invention relates to a pharmaceutical combination product comprising:(a) an inhibitor of the tyrosine kinase SRC; and(b) an autophagy inducer.Preferably, the inhibitor of the tyrosine kinase SRC is selected from the group consisting of:- SRC-specific miRNA inhibitors;- SRC-specific siRNA inhibitors;- SRC-specific proteolysis-targeting chimera (PROTAC);- dasatinib (A / -(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-1 -piperazinyl]- 2-methyl-4-pyrimidinyl]amino]-5-thiazole carboxamide), preferably in the form of monohydrate;- tirbanibulin (KX2-391 ; A / -Benzyl-2-(5-{4-[2-(4-morpholinyl)ethoxy]phenyl}-2- pyridinyl)acetamid);- bosutinib (SKI-606; 4-[(2,4-Dichlor-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4- methylpiperazin-1 -yl)propoxy]chinolin-3-carbonitrile);- ponatinib (AP24534; 3-(2-lmidazo[1 ,2-b]pyridazin-3-ylethynyl)-4-methyl- / \ / -(4- ((4-methylpiperazin-1 -yl)methyl)-3-(trifluoromethyl)phenyl)benzamide);- saracatinib (AZD0530; A / -(5-ch loro-1 ,3-benzodioxol-4-yl)-7-[2-(4-methyl-1 - piperazinyl)ethoxy]-5-[(tetrahydro-2 / - / -pyran-4-yl)oxy]-4-quinazolinamine); and- eCF506 (CAS No. 1914078-41 -3; N-[4-[4-amino-1 -[2-[4-(dimethylamino)-1 - piperidinyl]ethyl]-1 H-pyrazolo[3,4-d]pyrimidin-3-yl]-2-methoxyphenyl]-carbamic acid, 1 ,1 -dimethylethyl ester), or a pharmaceutically active salt or prodrug thereof; while the autophagy inducer is selected from the group consisting of:- Beclin-1 -specific activators;- SW076956 (CAS No. 851717-83-4; N-(4-(5-(furan-2-yl)-1 -(4-methylbenzoyl)- 4,5-dihydro-1 H-pyrazol-3-yl)phenyl)methanesulfonamide);- Tat-Beclin-1 or Tat-Beclin-1 -peptide;- SW063058 (CAS No. 533876-30-1 ; 7-Bromo-5-(4-fluorophenyl)-1 ,3,4,5- tetrahydro-4-(2-iodobenzoyl)-2H-1 ,4-benzodiazepin-2-one);- BRD1991 (CAS No. 2235468-02-5; 3,5-dichloro-N-((10-(1 -hydroxypropan-2-yl)- 8, 12-dimethyl-11 -oxo-7,8,9, 10,11 ,12-hexahydro-5H- benzo[9, 10]

[0001] oxa[5]azacycloundecino[7,8-b]indol-7-yl)methyl)-N- methylbenzamide);- BRD5631 (CAS No. CAS 2446154-91 -0; N-[(3S,6S,7S)-7-methoxy-3,6,9- trimethyl-10-oxo-3,4,5,6,7,8,9, 10-octahydro-2H-1 ,4,9- benzoxadiazacyclododecin-12-yl]-[1 ,1'-biphenyl]-4-carboxamide);- Tat-vFLIPa2; and- Tat-vFLIPa4, or a pharmaceutically active salt or prodrug thereof.In particularly effective embodiments of the first aspect,- (a) the inhibitor of the tyrosine kinase SRC is dasatinib (A / -(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-1-piperazinyl]-2-methyl- 4-pyrimidinyl]amino]-5-thiazole carboxamide), preferably in the form of monohydrate, and (b) the autophagy inducer is SW076956. SW076956 is designed to induce autophagy specifically, without affecting apoptosis, by targeting differences between the BH3 domain of Beclin 1 and those of molecules involved in cell death; or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW063058.Macroautophagy (herein also referred to as “autophagy”) is an intracellular degradation process, in which the engulfment of cytoplasmic and organelle components by double-membraned vesicles, autophagosomes, leads to their lysosomal degradation. Autophagy is an evolutionary conserved pathway that is implicated in multiple physiological and pathological conditions such as starvation, removal of damaged organelles, exercise, removal of intracellular pathogens and protection against diseases, such as cancer, respectively. Whether autophagy plays a cancer-protective role (possibly through the clearing of damaged organelles and protein aggregates and preventing DNA damage) in some cancer types while playing a cancer-promoting role (possibly by fueling the already established tumor) in others has been the subject of debate. Autophagy de-regulation, however, has been shown in several types of cancer, and it is now clear that it plays a role in both tumor formation and progression.Accordingly, in the embodiments of the present invention, the autophagy inducer is not an inhibitor of the tyrosine kinase SRC. As such, in embodiments where the inhibitor of the tyrosine kinase SRC is dasatinib or nintedanib, the autophagy inhibitor can neither be nintedanib nor dasatinib.Beclin-1 , also known as BECN1 (UniProtKB A0A024R1X5), the protein encoded by the BECN1 gene, is essential for autophagy. BECN1 is a haploinsufficient tumor suppressor, and although it is not known to be frequently mutated in cancers, becn1 + / - mice show an increased incidence of spontaneous tumors. BECN1 is part of the Class III PI3K complexes (PI3KC3) and is essential for vesicle enucleation (PI3KC3-C1 ) as well as for autophagosome maturation and endocytic trafficking (PI3KC3-C2). BECN1 is regulated at multiple levels, and is known to be phosphorylated by tyrosine kinases (TK).Importantly, the inventors have uncovered a new axis linking BAP1 loss with autophagy regulation through the non-receptor tyrosine kinase SRC and BECN1 , which provides a new targetable vulnerability in cancers with BAP1 loss.Accordingly, the pharmaceutical combination product according to the first aspect is particularly suitable for use in medical treatment of a patient, such as in cancertreatment and, in particular, in cancer treatment that targets BAP7-mutant and / or BAP1 -inactive cancer cells.As such, in further aspects, the present invention also relates to a method of treatment, in particular to a method of treatment of cancer such as of BAP7-mutant and / or BAP1 -inactive cancers, said method comprising administering to a subject:(a) an inhibitor of the tyrosine kinase SRC; and(b) an autophagy inducer.Typically, (a) the inhibitor of the tyrosine kinase SRC, and (b) the autophagy inducer are administered in the form of a combination product as defined herein.Similarly, in a further aspect, the present invention also relates to use of(a) an inhibitor of the tyrosine kinase SRC; and(b) an autophagy inducer, in the manufacture of a pharmaceutical combination product for medical treatment, in particular for the treatment of cancer such as of BAP7-mutant and / or BAP1 -inactive cancers.In the aspects relating to the medical treatment of a subject, (a) the inhibitor of the tyrosine kinase SRC; and (b) the autophagy inducer are administered in synergistically active dosages.Further, in a second aspect, the present invention relates to use of: (a) an inhibitor of the tyrosine kinase SRC; and (b) an autophagy inducer for manipulating BAP1 -mutant and / or BAP1 -inactive cells.Typically, (a) the inhibitor of the tyrosine kinase SRC, and (b) the autophagy inducer are used in the form of a combination product as defined herein.In embodiments of the second aspect, the inhibitor of the tyrosine kinase SRC and the autophagy inducer are used in synergistically active amounts such as to manipulate BAP7-mutant and / or BAP1 -inactive cells.Typically, the BAP1 -mutant or BAP 1 -inactive cells are manipulated in an in vitro assay for identifying treatment options for BAP7-mutant or BAP1 -inactive cancers.As outlined above, use of a combination of (a) an inhibitor of the tyrosine kinase SRC; and (b) an autophagy inducer, refers to use of two different active ingredients in combination.FIGURESEmbodiments of the invention will now be described, by way of example only, with reference to the accompanying drawings in which:Fig. 1 in accordance with the experiments described in Example 1 , illustrates the results of (a) reverse-phase protein array (RPPA) data analysis for SRC from KIRC-TCGA (ccRCC) stratified by BAP1 and / or PBRM1 mutation status; (b) RPPA data analysis from UVM-TCGA (UM) stratified by BAP1 mutations, (c) Western-blotting analysis of BAP 1 -deficient ccRCC UMRC-6 cell lines, (d) UM UPMM2 cell lines, (e) cholangiocarcinoma cell line TKF-1 and (f) breast carcinoma cell line HCC-1187, each reconstituted with an empty vector (EV), wild-type BAP1 or p.C91 S BAP1 mutant, (g) qRT-PCR analysis of reconstituted BAP 1 -deficient ccRCC line and (h) qRT-PCR analysis of reconstituted BAP1 -deficient UM cell line. **, P<0.01 ; ***, P<0.001 ; f-test.Fig 2. in accordance with the experiments described in Example 2, illustrates the results of autophagic flux of UMRC-6 cells reconstituted with an empty vector (EV) or BAP1 (wild-type or p.C91 S mutant) were analyzed by: (a) western blotting, (b) quantification of GFP-LC3 puncta and (c) quantification of HiBiT- LC3 luminescence, (d) GFP-LC3 puncta formation for UMRC-6 cells reconstituted with EV, wild-type BAP1 or several BAP1 mutants observed in tumors. The effects on autophagy of SRC inhibition were assessed by treatment with dasatinib and samples were analyzed by (e) western blotting, (f) HiBiT-LC3 luminescence, and (g) GFP-LC3 puncta under the microscope and its quantification (h), (i) GFP-FYVE puncta numbers of the indicated UMRC-6 cells treated with 1 pM dasatinib for 24h or DMSO control (UT), as well as through SRC silencing (j,k) or overexpression of a V5-tagged SRC construct (I). Bafilomycin A1 was added for 3 h at 100 nM and dasatinib at 1pM (or 5 pM were indicated) for 24 h. Error bars represent the average ± SEM of three independent experiments, n.s., non-significant. *, P<0.05; **, P<0.01 ; ***, P<0.001 for the indicated comparisons; f-test.Fig 3. in accordance with the experiments described in Example 3, illustrates the results of (a) BECN1 immunoprecipitation (IP) of UMRC-6 cells overexpressing SRC-V5 or an empty vector control (EV) containing plasmids for constitutively knocking down SRC (shSRC) or a scrambled control (shSc). (b) BECN1 (or IgG control) immunoprecipitation of UMRC-6 followed by western blotting analysis of phosphorylated tyrosine residues, (c) BECN1 immunoprecipitation of UMRC-6 cells overexpressing SRC-V5 (or an empty vector control, EV) along with BAP1 (or EV) treated with 1 pM dasatinib for 24 h or DMSO control (UT). (d-f) UMRC-6 cells stably expressing wild-type (WT) Flag-BECN1 , tyrosine phosphorylation mutants or an empty vector control (EV) were analyzed for autophagic flux by numbers of GFP-LC3 puncta (d), western blot (e) or cell proliferation (f). Error bars represent the average ± SEM of three independent experiments, n.s., non-significant. *, P<0.05; ***, P<0.001 compared to the negative control or the indicated comparisons; f-test.Fig 4. in accordance with the experiments described in Example 4, illustrates the effects on the induction of autophagy and cell viability in cells treated with a combination of an inhibitor of the tyrosine kinase SRC and an autophagy inducer: (a) combinatorial effects of dasatinib and SW076956 on autophagy induction in UMRC-6 cells assessed by western blotting, (b,c) combinatorial effects of SRC inhibitors dasatinib, saracatinib or bosutinib and autophagy inducer SW076956 in UMRC-6 cells assessed by HiBiT-LC3 luminescence or (d,e) numbers of GFP-LC3 puncta. Cells were treated with 1 pM dasatinib and / or 40 pM SW076956 for 24 h (a,b,d,e) or with 500 nM dasatinib, 10 pM saracatinib, 10 pM bosutinib and / or 40 pM SW076956 for 24 h (c). Error bars represent the average ± SEM of three independent experiments, n.s., nonsignificant, *, P<0.05; **, P<0.01 ; ***, P<0.001 compared to the negative control or the indicated comparisons; Ptest. Cell proliferation of UMRC-6 cells was assessed upon treatment with (f) 250 nM dasatinib (D) and / or 40 pM SW076956 (SW), (g) 30 pM of Tat-Beclin-1 -L11 S orTat-Beclin-1 peptide and / or 12.5 or 25 nM dasatinib or a DMSO control (Vehicle) or (h) two concentrations of rapamycin. Data represent the average± SEM of three independent experiments. *, P<0.05; **, P<0.01 ; ***, P<0.001 ; f-test. Combination effects of dasatinib and SW076956 treatment on cell viability of (i,j) UMRC-6 cells and (k ) TFK-1 cells, when reconstituted with an empty vector (EV), wild-type BAP1 or p.C91 S BAP1 mutant, were quantified 72 h after treatment of serial dilutions of single and combined compounds for three independent experiments. Synergy / antagonism effects of dasatinib and SW076956 were determined by a Loewe synergy model with Combenefit software of three independent experiments.Fig 5. in accordance with the experiments described in Example 4, illustrates the synergistic effects on the viability of cells treated with a combination of an inhibitor of the tyrosine kinase SRC and an autophagy inducer: (a,b) combinatorial effects of saracatinib and SW076956 on autophagy induction in UMRC-6 or TFK-1 cells, (c,d) combinatorial effects of bosutinib and SW076956 on autophagy induction in UMRC-6 or TFK-1 cells. Cells were were quantified 72 h after treatment with serial dilutions of single and combined compounds. Synergy / antagonism effects of saracatinib or bosutinib and SW076956 were determined by a Loewe synergy model with Combenefit software of three independent experiments in UMRC-6 cells (a,c) and four independent experiments in TFK-1 cells (b,d). *, P<0.05; **, P<0.01 ; ***, P<0.001.Fig 6. in accordance with the experiments described in Example 4, illustrates the synergistic effects on the viability of cells treated with a combination of an inhibitor of the tyrosine kinase SRC and an autophagy inducer in the context of BAP1 loss: Combination effects of dasatinib and SW063058 treatment (a) or of saracatinib and SW063058 treatment (b) on cell viability of TFK-1 cells were quantified 72 h after treatment of serial dilutions of single and combined compounds in at least three independent experiments. Synergy / antagonism effects of dasatinib or saracatinib and SW063058 were determined by a Loewe synergy model with Combenefit software of three independent experiments. *, P<0.05; **, P<0.01 ; ***, P<0.001.Fig 7. in accordance with the experiments described in Example 5, illustrates the results of (a) chorioallantoic membrane (CAM) assay for representative tumors and (b) quantification of tumor growth of UMRC-6 cells implanted in fertilized chick eggs and treated with 1 pM dasatinib and / or 40 pM SW076956 for 7 days. Tumors generated from a CAM assay of UMRC-6 cells treatedwith Tat-BECN1 peptide (TB) or Tat-Scrambled control (TS) (i.e. Tat-Beclin- 1 -L11S at 10 mM for 3 h) were analyzed (c,d). *, P<0.05; **, P<0.01 ; f-test.Fig 8. in accordance with the experiments described in Example 6, illustrates the combination effects of dasatinib and SW076956 treatments on cell viability of patient-derived tumor organoids (PDTOs) from a ccRCC tumor (a) with BAP1 loss and (b) with wild-type BAP1 as well as of PDTOs from a UM tumor (c) with BAP1 loss and (d) with wild-type BAP1. (e) Proposed mechanism of regulation of SRC and autophagy by BAP1 loss through the binding and phosphorylation of BECN1 by SRC and rationale for treating BAP1 -mutant cancers with SRC inhibitors and autophagy inducers.DETAILED DESCRIPTION OF THE INVENTIONIn order to provide a clear and consistent understanding of the specification and claims, and the scope to be given such terms, the following definitions are provided.DefinitionsThe term “pharmaceutical combination product” in the context of this disclosure has its conventional meaning in that at least two different active ingredients being used in combination such that it refers either to a product in which a single composition comprises both active ingredients (a) an inhibitor of the tyrosine kinase SRC and (b) an autophagy inducer, or to a product, such as a kit, in which the two active ingredients (a) an inhibitor of the tyrosine kinase SRC and (b) an autophagy inducer are provided together but in separate compositions, possibly even for simultaneous or sequential administration via the same or a different route of administration.In the context of this disclosure, the term “inhibitor of the tyrosine kinase SRC” means a compound that blocks the catalytic activity of the tyrosine kinase SRC. For instance, it can compete with adenosine triphosphate (ATP), which is involved in adding phosphate groups, either as the substrate or as the phosphorylating entity. Alternatively, it can work in a different way by binding to a different site on the enzyme, called an allosteric site, and affecting its activity by causing a conformational change.Some FDA-approved drugs like lithium, metformin and statins usually have multiple, often non-specific effects, one of which can be to trigger autophagy. However, given the multiple effects of such drugs, it is difficult to determine whether any beneficial effects seen in patients are the consequence of the activation of the autophagy machinery or of other mechanisms at play.Therefore, when identifying compounds that specifically induce autophagy, it is crucial to prove that a compound’s effects are indeed a direct consequence of the activation of the autophagy machinery. To illustrate such a direct and specific effect, compounds that specifically induce autophagy are unable to elicit their effects when the autophagy machinery is compromised, i.e. non-functional. In turn, non-specific signals that activate autophagy can also affect other pathways, such as the mammalian target of rapamycin complex 1 (mTORCI ), the 5' AMP-activated protein kinase (AMPK) and protein kinase B (AKT) pathways. Therefore, it is important to distinguish between effects achieved because of the activation of autophagy or because of the modification of other pathways. For example, BH3 mimetic compounds disrupt the interaction between Beclin 1 and Beclin 1 inhibitors of the B-cell lymphoma 2 (Bcl-2) protein family and increase autophagy. However, such compounds are not autophagy inducers as understood in the context of the present disclosure as they also promote apoptosis by interrupting interactions between Bcl-2 family members and BH3 domains of molecules linked to apoptosis.Specific autophagy inducing agents target regulators of the autophagy process or parts of the molecular autophagy machinery itself to specifically induce autophagy without affecting other processes, e.g. without affecting apoptosis and / or without inhibiting the catalytic activity of the tyrosine kinase SRC.Therefore, in the context of this disclosure, the term “autophagy inducer” means a compound that is able to promote autophagy flux by targeting regulators of the autophagy process and / or parts of the molecular autophagy machinery itself to specifically induce autophagy but without affecting other processes, e.g. without affecting apoptosis. Therefore, the term encompasses compounds, which (A) target the binding of Beclin 1 to Bcl-2 without affecting apoptosis (such as, for example, SW076956, SW063058, BRD1991 and BRD5631 ), or (B) which target the binding of Beclin-1 to GAPR-1 / GLIPR2 without affecting apoptosis (such as, for example, Tat-Beclin-1 peptide, Tat-Beclin-1 -D11 or Tat-Beclin-1 -L11 ); or (C) which target the binding of ATG3 to LC3 (such as, for example Tat-vFLIPa2 or Tat-vFLIPa4).Importantly, an autophagy inducer in the context of the present invention does not inhibit the catalytic activity of the tyrosine kinase SRC. Pharmaceutically active ingredients that inhibit SRC and, coincidently, also induce autophagy are expressly excluded from the scope of the term “autophagy inducer” as used throughout this disclosure.The term “SRC-specific miRNA inhibitors” in the context of this disclosure refers to microRNA (miRNA) molecules, which selectively via SRC-sequence-specificity decrease SRC protein production through translational attenuation and / or SRC messenger RNA (mRNA) degradation.The term “SRC-specific siRNA inhibitors” in the context of this disclosure refers small-interfering RNA (siRNA) molecules, which selectively via SRC-sequence- specificity decrease SRC protein production through translational attenuation and / or SRC messenger RNA (mRNA) degradation.In the context of this disclosure, the term “SRC-specific proteolysis-targeting chimera” or “SRC-specific PROTAC” means a heterobifunctional molecule composed of two active domains and a linker, capable of specifically removing SRC. Rather than acting as a conventional SRC enzyme inhibitor, a SRC-specific PROTAC induces selective intracellular proteolysis of SRC. PROTACs consist of two covalently linked protein-binding molecules: one capable of engaging an E3 ubiquitin ligase, and another that binds to a target protein meant for degradation. Therefore, through SRC- specific PROTACs a cell's ubiquitin-proteasome system (UPS) may be “hijacked” by recruiting the E3 ubiquitin ligase to the SRC protein such that SRC ubiquitination and subsequent degradation via the proteasome is affected.The term “Beclin-1 -specific activators” in the context of this disclosure refers to autophagy inducers that induce autophagy via regulation of Beclin-1 , for example, by post-translational modification (such as phosphorylation or acetylation), by modification of its interactome, or by changes to its subcellular localization.The term “Tat-peptide” or “Tat-” in the context of this disclosure refers to a cell penetrating peptide derived from the HIV-1 trans-activating transcriptional activator (Tat; UniProtKB: P35965) having the sequence of SEQ ID NO: 1 X1GRKKRRQRRRX2X3) or the retro inverse sequence of SEQ ID NO: 2 X3X2RRRQRRKKRGX1, where Xi is either Y or absent, X2 is either P or absent and X3 is either Q or absent.The term “Tat-Beclin-1” in the context of this disclosure refers to an N-terminal fusion of Tat-peptide, typically via a linker, preferably via a di-glycine linker, to full-length Beclin-1 protein (SEQ ID NO: 3) or a variant thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO: 3. For example, a variant of full-length Beclin-1 protein of SEQ ID NO: 3 may be SEQ ID NO: 4 or SEQ ID NO: 5 or SEQ ID NO: 6 or SEQ ID NO: 7 or SEQ ID NO: 8). Importantly, in the context of the present invention, Tat-Beclin-1 and variants thereof maintain Beclin-1 function, namely Beclin-1 function relating to vesicle enucleation (PI3KC3-C1 ) as well as to autophagosome maturation and endocytic trafficking.In the context of this disclosure, the term “Tat-Beclin-1 peptide" means a peptide comprising: (1 ) Tat-peptide of SEQ ID NO: 1 or SEQ ID NO: 2; (2) a linker sequence, preferably a di-glycine linker; and (3) a Beclin-1 sequence comprising amino acid residues 267-284 of the autophagy protein Beclin-1 (SEQ ID NO: 9 TNVFNATFHIWHSGQFGT) or variants thereof having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO: 9. A particularly useful variant of the Beclin-1 sequence of (3) above is the peptide variant of SEQ ID NO: 10 (TNVFNATFEIWHDGEFGT; having 83.33% sequence identity to SEQ ID NO: 9). The peptides of SEQ ID NO: 11 or SEQ ID NO: 12 are express examples of Tat-Beclin-1 peptides encompassed by the above term.Tat-Beclin-1 -peptides are cell-penetrating peptides demonstrated to induce autophagy through interaction with the autophagy suppressor GAPR-1 / GLIPR2 both in vitro and in vivo. Upon peptide binding, Beclin-1 bound to GARP-1 is released, resulting in Beclin-1 mediated autophagosome formation and autophagy induction. Illustrative of its specificity, i.e. as an autophagy inducer as understood herein, it isnoteworthy that Tat-Beclin-1 peptides cannot affect autophagosome formation and autophagy induction in situations where sufficient amounts of Beclin 1 are lacking.The term “Tat-Beclin-1 -D11” in the context of this disclosure refers to a shorter, reengineered variant of the Tat-Beclin-1 peptide, only including 11 AAs of Beclin-1 (NBP2-49888; SEQ ID NO: 13 RRRQRRKKRGYGGDHWIHFTANWV), which demonstrates enhanced autophagy induction compared to the Tat-Beclin 1 peptide.The term “Tat-Beclin-1 -L11” in the context of this disclosure refers to a shorter, reengineered variant of the Tat-Beclin-1 peptide, only including 11 AAs of Beclin-1 (NBP2-49886; SEQ ID NO: 14 YGRKKRRQRRRGGVWNATFHIWHD), which demonstrates enhanced autophagy induction compared to the Tat-Beclin 1 peptide.The term “Tat-Beclin-1 -L11S” in the context of this disclosure also refers to a shorter, re-engineered inactive / scrambled variant of the Tat-Beclin-1 peptide (NBP2- 49887; SEQ ID NO: 15 YGRKKRRQRRRGGNWAWHDFVHIT), which is useful as a negative control.Both Tat-Beclin-1 -D11 and Tat-Beclin-1 -L11 peptides function by binding the negative regulator of autophagy GAPR-1 / GLIPR2. Upon peptide binding, Beclin-1 bound to GARP-1 is released, resulting in Beclin-1 mediated autophagosome formation and autophagy induction.The term “Tat-vFLIPa2” in the context of this disclosure refers to an N-terminal fusion of Tat-peptide to amino acids 20-29 (SEQ ID NO: 16 FVNLLFLWE) of full length viral FLICE-like inhibitor protein (SEQ ID NO: 17 and 18, respectively).The term “Tat-vFLIPa4” in the context of this disclosure refers to an N-terminal fusion of Tat-peptide to amino acids 128-139 (SEQ ID NO: 19 NEMCYVWHLFTQ) of full length viral FLICE-like inhibitor protein (SEQ ID NO: 20 and 21 , respectively).Both Tat-vFLIPa2 and Tat-vFLIPa4 bind to the FLIP protein itself preventing its binding to autophagy related 3 (ATG3) and, in turn, allowing the binding of ATG3 to LC3 and thereby promoting the autophagosome formation and, as a consequence, the induction of autophagy.In the context of this disclosure, the term “BAPl-mutant or BAP1 -inactivate cancer cells” refers to cancer cells, in particular to in vivo tumor cells as well as to in vitro cancer cell lines, which lack active BAP1 protein, i.e. to cells with BAP1 loss. Specifically, the term does not distinguish between this lack or loss of BAP1 protein being the result of a mutation at the level of the gene, which results in a lack of any translatable BAP1 transcript, or a lack of functionality due to the production of misfolded, truncated or otherwise inactive BAP1 proteins. Instead, the term is to be understood to encompass all cancer cells that lack active BAP1 protein.In addition to the above definitions, and unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.Further, reference throughout this specification to “one embodiment”, “some embodiments” or “an embodiment” means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases “in one embodiment”, “in some embodiments” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment, but may. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as would be apparent to one of ordinary skill in the art from this disclosure, in one or more embodiments.As used herein, unless otherwise specified the use of the ordinal adjectives "first", "second", "third", etc., to describe a common object, merely indicate that different instances of like objects are being referred to, and are not intended to imply that the objects so described must be in a given sequence, either temporally, spatially, in ranking, or in any other manner.As used herein, the term “exemplary” is used in the sense of providing examples, as opposed to indicating quality. That is, an “exemplary embodiment” is an embodiment provided as an example, as opposed to necessarily being an embodiment of exemplary quality.Embodiments of the inventionAs indicated above, the present invention is based on the inventors’ surprising finding that pharmaceutical combination products comprising two different active ingredients: (a) an inhibitor of the tyrosine kinase SRC and (b) an inducer of autophagy are effective in the treatment of cancer, in particular of cancers with BAP1 loss.Macroautophagy (herein also referred to as “autophagy”) is an intracellular degradation process, in which the engulfment of cytoplasmic and organelle components by double-membraned vesicles, autophagosomes, leads to their lysosomal degradation. Autophagy is an evolutionary conserved pathway that is implicated in multiple physiological and pathological conditions such as starvation, removal of damaged organelles, exercise, removal of intracellular pathogens and protection against diseases, such as cancer, respectively. BECN1 , also known as Beclin-1 (BECN1 ), the protein encoded by the BECN1 gene, is essential for autophagy. BECN1 is a haploinsufficient tumor suppressor, and although is not frequently mutated in cancers, becn1 + / - mice show an increased incidence of spontaneous tumors. BECN1 is part of the Class III PI3K complexes (PI3KC3) and it is essential for vesicle enucleation (PI3KC3-C1 ) as well as autophagosome maturation and endocytic trafficking (PI3KC3-C2). Autophagy de-regulation has been shown in several types of cancer, and it is now clear that it plays a role in both tumor formation and progression. BECN1 itself does not have catalytic activity but is required for activation of the PI3KC3-C1 complex, containing the kinase VPS34. This complex is essential for the formation of the autophagosome and therefore essential for autophagy. Beclin-1 modulates the activity of this complex. Whether autophagy plays a “protecting” or a “promoting” role in cancer needs to be determined for each cancer type and stage. BECN1 is regulated at multiple levels, and it was shown to be phosphorylated by tyrosine kinases (TK) implicated in tumorigenesis, such as HER2, EGFR or BCR-ABL.BAP1 is a tumor suppressor and epigenetic modifier that is frequently mutated in multiple cancers, such as uveal melanoma (UM), malignant pleural mesothelioma (MPM), cutaneous melanoma, renal cell carcinoma (RCC), cholangiocarcinoma, basal cell carcinoma, and likely other malignancies, leading to higher tumor aggressiveness, metastasis and poor patient survival.The inventors have found that BAP1 transcriptionally regulates protooncogene SRC, a non-receptor tyrosine kinase, and that SRC binds, phosphorylates and inactivates BECN1 (an essential autophagy protein and haploinsufficient tumor suppressor) in cells derived from multiple cancer entities. Furthermore, proliferation of these cells is dependent on the BECN1 phosphorylation status but can be modulated using a pharmaceutical combination product in accordance with the present invention, which provides a synergistic combination of (a) a SRC inhibitor and (b) an autophagy inducer.As such, a new axis linking BAP1 loss with autophagy regulation through the nonreceptor tyrosine kinase SRC and BECN1 , which provides a new targetable vulnerability in cancers, in particular in cancers with BAP1 loss, is described.In fact - for the first time, it is described that administration of an inhibitor of the tyrosine kinase SRC in combination with an autophagy inducer (i.e. a combination of two different active ingredients with the respective functionalities) can potentiate the autophagy inducer’s effects to restore de-regulated autophagy and, thereby, to counteract both tumor formation and progression, in a synergistic manner.Accordingly, in a first aspect, the present invention relates to a pharmaceutical combination product comprising: (a) an inhibitor of the tyrosine kinase SRC; and (b) an autophagy inducer.Preferably, the inhibitor of the tyrosine kinase SRC used in the pharmaceutical combination product of the present invention is selected from the group consisting of:- SRC-specific miRNA inhibitors;- SRC-specific siRNA inhibitors;- SRC-specific proteolysis-targeting chimera (PROTAC);- dasatinib (A / -(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-1 -piperazinyl]- 2-methyl-4-pyrimidinyl]amino]-5-thiazole carboxamide), preferably in the form of monohydrate;- tirbanibulin (KX2-391 ; A / -Benzyl-2-(5-{4-[2-(4-morpholinyl)ethoxy]phenyl}-2- pyridinyl)acetamid);- bosutinib (SKI-606; 4-[(2,4-Dichlor-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4- methylpiperazin-1-yl)propoxy]chinolin-3-carbonitrile);- ponatinib (AP24534; 3-(2-lmidazo[1 ,2-b]pyridazin-3-ylethynyl)-4-methyl- / \ / -(4- ((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide);- saracatinib (AZD0530; A / -(5-ch loro-1 ,3-benzodioxol-4-yl)-7-[2-(4-methyl-1- piperazinyl)ethoxy]-5-[(tetrahydro-2H-pyran-4-yl)oxy]-4-quinazolinamine); and- eCF506 (CAS No. 1914078-41-3; N-[4-[4-amino-1-[2-[4-(dimethylamino)-1- piperidinyl]ethyl]-1 H-pyrazolo[3,4-d]pyrimidin-3-yl]-2-methoxyphenyl]-carbamic acid, 1 ,1 -dimethylethyl ester), or a pharmaceutically active salt or prodrug thereof, while the autophagy inducer is selected from the group consisting of:- Beclin-1 -specific activators;- SW076956 (CAS No. 851717-83-4; N-(4-(5-(furan-2-yl)-1-(4-methylbenzoyl)- 4,5-dihydro-1 H-pyrazol-3-yl)phenyl)methanesulfonamide);- Tat-Beclin-1 ;- Tat-Beclin-1 peptide;- Tat-Beclin-1 -D11 peptide;- Tat-Beclin-1 -L11 peptide;- SW063058 (CAS No. 533876-30-1 ; 7-Bromo-5-(4-fluorophenyl)-1 ,3,4,5- tetrahydro-4-(2-iodobenzoyl)-2H-1 ,4-benzodiazepin-2-one);- BRD1991 (CAS No. 2235468-02-5; 3,5-dichloro-N-((10-(1 -hydroxypropan-2-yl)- 8, 12-dimethyl-11 -oxo-7,8,9, 10,11 ,12-hexahydro-5H- benzo[9, 10]

[0001] oxa[5]azacycloundecino[7,8-b]indol-7-yl)methyl)-N- methylbenzamide);- BRD5631 (CAS No. CAS 2446154-91 -0; N-[(3S,6S,7S)-7-methoxy-3,6,9- trimethyl-10-oxo-3,4,5,6,7,8,9, 10-octahydro-2H-1 ,4,9- benzoxadiazacyclododecin-12-yl]-[1 ,1'-biphenyl]-4-carboxamide);- Tat-vFLIPa2; and- Tat-vFLIPa4, or a pharmaceutically active salt or prodrug thereof.In some embodiments,- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is SW076956 (SW076956 is designed to induce autophagy specifically, without affecting apoptosis, bytargeting differences between the BH3 domain of Beclin 1 and those of molecules involved in cell death): or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW063058.In aspects of the present invention relating to the medical treatment of a subject, (a) the inhibitor of the tyrosine kinase SRC; and (b) the autophagy inducer are administered in synergistically active dosages.For example, the daily dosage of the inhibitor of the tyrosine kinase SRC administered to a subject in need thereof via the pharmaceutical combination product of the present disclosure ranges between 0.1 milligram per kilogram bodyweight (mg / kg) and 20 mg / kg, such as between 0.1 mg / kg and 17.5 mg / kg or between 0.1 mg / kg and 15 mg / kg or between 0.1 mg / kg and 12.5 mg / kg or between 0.1 mg / kg and 10 mg / kg or between 0.1 mg / kg and 7.5 mg / kg or between 0.1 mg / kg and 5 mg / kg or between 0.1 mg / kg and 2.5 mg / kg or between 0.1 mg / kg and 2 mg / kg or between 0.1 mg / kg and1.5 mg / kg or between 1 mg / kg and 7.5 mg / kg or between 1 mg / kg and 5 mg / kg, preferably the daily dosage is about 0.1 mg / kg or about 0.25 mg / kg or about 0.5 mg / kg or about 0.75 mg / kg or about 1 mg / kg or about 1 .5 mg / kg or about 2 mg / kg or about2.5 mg / kg or about 3 mg / kg or about 3.5 mg / kg or about 4 mg / kg or about 4.5 mg / kg or about 5 mg / kg or about 5.5 mg / kg or about 6 mg / kg.Similarly, the daily dosage of the autophagy inducer administered to a subject in need thereof via the pharmaceutical combination product of the present disclosure ranges between 0.1 milligram per kilogram bodyweight (mg / kg) and 100 mg / kg, such as between 0.1 mg / kg and 90 mg / kg or such as between 0.1 mg / kg and 80 mg / kg or such as between 0.1 mg / kg and 70 mg / kg or such as between 0.1 mg / kg and 60 mg / kg or such as between 0.1 mg / kg and 50 mg / kg or such as between 0.1 mg / kg and 40 mg / kg or such as between 0.1 mg / kg and 30 mg / kg or such as between 0.1 mg / kg and 20 mg / kg or such as between 0.1 mg / kg and 17.5 mg / kg or between 0.1 mg / kg and 15 mg / kg or between 0.1 mg / kg and 12.5 mg / kg or between 0.1 mg / kg and 10 mg / kg or between 0.1 mg / kg and 7.5 mg / kg or between 0.1 mg / kg and 5 mg / kg or between 0.1 mg / kg and 2.5 mg / kg or between 0.1 mg / kg and 2 mg / kg or between 0.1 mg / kg and 1.5 mg / kg or between 1 mg / kg and 7.5 mg / kg or between 1 mg / kg and 5 mg / kg, preferably the daily dosage is about 0.1 mg / kg or about 0.25 mg / kg or about 0.5 mg / kg or about 0.75 mg / kg or about 1 mg / kg or about 1 .5 mg / kg or about 2 mg / kg or about 2.5 mg / kg or about 3 mg / kg or about 3.5 mg / kg or about 4 mg / kg or about 4.5 mg / kg or about 5 mg / kg or about 5.5 mg / kg or about 6 mg / kg.Importantly, the pharmaceutical combination product of the present invention may comprise (a) the inhibitor of the tyrosine kinase SRC and (b) the autophagy inducer, as separate entities. As such, the combination product may be a kit, preferably for use in medical treatment such as cancer treatment, comprising a SRC tyrosine kinase inhibitor composition and a separate autophagy inducer composition for simultaneous or sequential administration.In this regard, it is noteworthy that the two separate entities of a pharmaceutical combination product of the invention (i.e. such as a kit) may be formulated for the same or for distinct routes of administration. For example, the SRC tyrosine kinase inhibitor composition may be formulated for intratumoral or intraperitoneal or systemic or intranasal or oral administration, while the autophagy inducer composition may be formulated for intratumoral or intraperitoneal or systemic or intranasal or oral administration. In preferred embodiments, (a) the inhibitor of the tyrosine kinase SRC is formulated for oral administration and (b) the autophagy inducer is formulated for intraperitoneal administration.In alternative embodiments, the pharmaceutical combination product of the present invention may comprise (a) the inhibitor of the tyrosine kinase SRC; and (b) the autophagy inducer, in a single composition. For example, both (a) and (b) may be formulated for joint oral or intraperitoneal or systemic administration.In any case, in all of the above embodiments of the present invention, the autophagy inducer is not an inhibitor of the tyrosine kinase SRC. As such, in embodiments where the pharmaceutical combination product comprises either dasatinib or nintedanib as (a) the inhibitor of the tyrosine kinase SRC is, the autophagy inhibitor can neither be nintedanib nor dasatinib.In ovo experiments confirm that tumor growth in BAP 1 -deficient cells is decreased when treated with a pharmaceutical combination product in accordance with the present invention. In addition, studies both in vitro in multiple cell lines, as well as ex vivo in patient-derived tumor organoids (PDTOs) from UM and ccRCC as preclinical models, confirm the synergistic effect of a SRC inhibitor and an autophagy inducer.Therefore, it will be understood from the above but also from the Examples and associated Figures of the present disclosure that the pharmaceutical combination product of the first aspect is particularly suitable for use in medical treatment of a patient, such as in cancer treatment and, in particular, in cancer treatment that targets BAP7-mutant and / or BAP1 -inactive cancer cells.In some of these embodiments, the pharmaceutical combination product for use according to the invention allows for treatment of a cancer or tumor by targeting BAP1- mutant or BAP1 -inactive cancer cells.The cancer treatment in this context (and as expressly defined above) includes the treatment of melanoma, renal cancer, pleural cancer and / or gallbladder cancer. In particular, the pharmaceutical combination product of the present invention is particularly suitable for use in the treatment of uveal melanoma (UM), clear-cell renal cell carcinoma (ccRCC), malignant pleural mesothelioma (MPM), and / or intrahepatic cholangiocarcinoma.When using the pharmaceutical combination product of the present invention in the above-described treatments, this increases the cancer’s susceptibility to autophagy activation. As explained above, the inventors’ found that the administration of the pharmaceutical combination product in accordance with the above embodiments of the invention disrupts the oncogenic / deregulated SRC tyrosine kinase binding to and phosphorylation of BECN1. Without wanting to be bound by theory, it is this effect, which allows for the re-activation of the deregulated and suppressed autophagy machinery in the tumor and, which allows for the SRC tyrosine kinase inhibitor and the autophagy inducer to act synergistically and to counteract the detrimental effects of autophagy de-regulation in these cancer entities.In particular, treatment with the pharmaceutical combination product of the invention reduces the cancer’s growth and, preferably, reduces tumor size.As indicated above, the present invention in a second aspect also relates to the combinatorial use of (a) an inhibitor of the tyrosine kinase SRC; and (b) an autophagy inducer, for manipulating BAP1 -mutant or BAP1 -inactive cells.In embodiments of the second aspect, the inhibitor of the tyrosine kinase SRC and the autophagy inducer are used in synergistically active amounts such as to manipulate BAP7-mutant and / or BAP1 -inactive cells.Generally, the BAP7-mutant or BAP1 -inactive cells are manipulated such that Beclinl (BECN1 ) phosphorylation is decreased. In such instances, the BAPPmutant or BAP1 -inactive cells are manipulated such that their proliferation is decreased.As the skilled person will appreciate, such manipulations are typically performed in vitro, in an in vitro assay, and may provide useful tools for the analysis of BAP1- mutant or BAP1 -inactive cells, providing further insights into vulnerabilities of such cells, which may be exploited in the quest for identifying further treatment options for BAP7-mutant or BAP1 -inactive cancers.The invention is further described by the following non-limiting Examples.Methods and materialsCell culture, antibodies and reac / entsThe ccRCC cell line with a BAP1 frameshift deletion, UMRC-6 (or UM-RC-6), was obtained from Sigma-Aldrich. The cholangiocarcinoma cell line with a BAP1 nonsense mutation, TFK-1 , was a generous gift from Dr. Stephanie Rdssler (University Hospital Heidelberg). The primary uveal melanoma cell line UPMM2 with a BAP1 mutation was kindly provided as early passage by Dr. Michael Zeschnigk (University Hospital Essen). The primary breast cancer cell line with a nonsense BAP1 mutation, HCC- 1187, was a gift from Prof. Adi Gazdar (UT Southwestern Medical Center). The ccRCC cell line with wild-type BAP1 , 786-0, was obtained from ATCC. HEK-293T cells were kindly provided by Prof. Stefan Frdhling (DKFZ, Heidelberg). All cell lines were maintained in their corresponding medium supplemented with 10% (v / v) fetal bovine serum (FBS) (Gibco, 10500064) and 1 % (v / v) Penicillin-Streptomycin (Gibco, 15140122) in a humidified incubator at 37 °C and 5% CO2. The growth medium for UMRC-6 and HEK-293T cells was Dulbecco's Modified Eagle’s Medium (DMEM) (Gibco, 41965062), for TFK-1 , HCC-1187 and 786-0 cells was RPMI (Gibco, 61870044) and for UPMM2 was Ham’s F-12 (Gibco, 31765068). Cells were routinely tested for mycoplasma by PCR. Cell lines were authenticated using Multiplex Cell Authentication by Multiplexion (Heidelberg, Germany).SRC inhibitor dasatinib was purchased from Biomol and dissolved in DMSO to a stock concentration of 10 mM. Further dilutions in DMSO were made prior to use. Additional SRC inhibitors, bosutinib and saracatinib, were obtained from Biocat and Santa Cruz, respectively, and dissolved at 10 mM in DMSO. Bafilomycin A1 was from MedChemExpress and diluted to 1 mM in DMSO. Tat-Beclin-1 -D11 peptide (NBP2- 49888) and Tat-Beclin-1 -L11 S scrambled peptide (NBP2-49887) were purchased from Novus-Biologicals. SW076956 was obtained from D&C Chemicals (Shanghai, China) and dissolved at 50 mM in DMSO. SW063068 was purchased from Merck and dissolved at 2 mM in DMSO.Antibodies were from the following sources: Santa Cruz (BECN1 for IP (mouse E-8), Actin-HRP, total-phospho-Tyr-HRP, HRP-conjugated secondary antibodies), Millipore (SRC), Cell Signalling Technology (BECN1 #3738 for WB, phospho-SRC #2101 [Tyr416 for mouse, Tyr419 for human], p62 #88588), Invitrogen (V5), Echelon (VPS34).PlasmidsV5-tagged (C-terminal) SRC (pLX304-SRC-V5) was created by Gateway recombination of lentiviral vector pLX304 (Addgene #25890) with pENTR223-SRC (ClonelD: 170040104) obtained from the Gateway Full ORF Clones at the DKFZ Vector and Clone Repository. The plasmids to stably express wild-type BAP1 (pBABE-hygro-BAP1 -HA, Addgene #154020), catalytically-inactive p.C91 S mutant BAP1 (pBABE-hygro-BAP1 -C91 S-HA, Addgene #154021 ) or an empty vector control (pBABE-hygro, Addgene #1765) were used. Additional BAP1 plasmids were kindly provided by Prof. Boyi Gan: pLVX-M-puro, pLVX-Flag-BAP1 , p.C91A, p.S10T, p.E31A, p.L49V, p.H169Q and p.G185R (Addgene #125839-4, 125847, 125849). The pBABE-puro-GFP-LC3 and Flag-BECN1 (WT, Y229 / 233 / 352F, Y229 / 233 / 352E) plasmids were used. HiBiT-LC3 reporter vector was from Promega. Lentiviral pLKO.1 short-hairpin RNA (shRNA) SRC vectors (TRCN0000038150 and TRCN0000199313) and scrambled control (SHC002) were from Sigma.Cell line reconstitution with BAP1 constructsThe BAP 1 -deficient cell lines UMRC-6, TFK-1 , and UPMM2 were reconstituted with wild-type BAP1 (pBABE-hygro-BAP1 -HA, Addgene #154020), catalytically-inactive p.C91 S mutant BAP1 (pBABE-hygro-BAP1 -C91 S-HA, Addgene #154021 ) or an empty vector control (pBABE-hygro, Addgene #1765). Additional validation was established by transducing the cell lines with wild-type BAP1 (pLVX-Flag-BAP1 , Addgene #125840), several mutant BAP1 constructs (p.C91A, p.S10T, p.E31A, p.L49V, p.H169Q and p.G185R, Addgene #125841 -4, 125847, 125849) or an empty vector control (pLVX-M-puro, Addgene #125839).Lentiviral Production and Transduction of VectorsFor stable SRC overexpression and / or SRC knockdown, lentiviruses were generated in HEK-293T cells cultured in DMEM medium supplemented with 10% FBS and 1 % Penicillin-Streptomycin. HEK-293T cells were transfected with pMD2.G envelope plasmid (Addgene #12259), psPAX2 packaging plasmid (Addgene #12260) and pLKO.1 vectors containing shRNA sequences against SRC. As control, pLKO.1 - Scrambled was used. Virus-containing supernatant was collected and passed through a 0.45 pm syringe filter (Coming, 431220). Cell culture medium of cell lineswas replaced with filtered virus-containing supernatant for transduction and cells were selected with 2 pg / ml puromycin (Life Technologies, A1 113803). siRNA and plasmid transfectionsDicer siRNA oligos were obtained from IDT (Integrated DNA Technologies) and transfected with siQUEST (Mirus) according to the manufacturer’s instructions. Assays were performed two days after siRNA transfection. Plasmids were transfected using the Trans-LT1 (Mirus) transfection reagent according to the manufacturer’s instructions.Generation of HiBiT-LC3 and Flag-BECN1 cell linesUMRC-6 cells were seeded into 10 cm2dishes and transfected 24 h later using the corresponding vector: autophagy HiBiT-LC3 reporter vector or Flag-BECN1 (WT and mutants p.Y229 / 233 / 352F, p.Y229 / 233 / 352E or empty vector control). Two days after transfection cells were selected with 250 pg / ml G418 (250 pg / ml) until all cells died in the control, mock-transfected plate. Cells were maintained and expanded in G418 and experiments were performed without G418.CRISPR / Cas9 knockout of BAP1 in 786-0 cellsBAP1 gene was knocked out in the BAP 1 -com petent 786-0 cells using Synthego’s Gene Knockout Kit (Redwood City, CA, USA). Briefly, 786-0 cells were electroporated with sgRNA-Cas9 complexes targeting BAP1.Namely with:- SgRNA2 BAP 1 +52408564: Ums-Cms-Ams-Aro-Aro-Uro-Gro-Gro-Aro-Uro-Cro-Gro-Aro- Aro-Gro-Aro-Gro-Cro-Gro-Cro (SEQ ID NO: 24) + Synthego modified EZ scaffold;- sgRNA4 BAP 1 +52408601 : Gms-Gms-Ams-Aro-Gro-Aro-Uro-Aro-Aro-Aro-Uro-Cro-Cro- Aro-Uro-Aro-Uro-Aro-Cro-Aro (SEQ ID NO: 25) + Synthego modified EZ scaffold; or- Cas9 control using 4D-Nucleofector X Kit (Lonza, V4XC-2032).In the above shown nucleotide sequences targeting BAP1 , i.e. in sgRNA2 BAP1 and sgRNA4 BAP1 , “ms” indicates that the respective nucleotides are O-methylated and have a phosphothiolated backbone structure (P=S), wherein “ro” indicates that tertbutyldimethylsilyl (TBDMS) protecting groups are attached to hydroxyl groups of therespective nucleotides, while the regular phosphodiester backbone structure (P=O) is maintained.Electroporated cells were separated as single cells by limiting dilution and grown into single clones through several rounds of clonal expansion. BAP1 nucleotide deletion was verified by Sanger sequencing and BAP1 loss by western blotting as described below.Cell lysates, immunoprecipitation and western blottingCells were washed in ice-cold PBS and lysed in ice-cold lysis buffer (Tris-HCI pH7.4, 50 mM, NaCI 250 mM, Igepal 0,5%) containing protease inhibitors (complete Protease Inhibitor Cocktail, Roche or Thermo Fisher Scientific, 11834101 ) and phosphatase inhibitors (PhosSTOP Phosphatase Inhibitor Cocktail, Roche or Thermo Fisher Scientific, 11814101 ) for 10 minutes at 4 °C. Cell debris was depleted by centrifugation at 16,000 g for 10 min at 4 °C and protein concentration was assessed by Bradford assay. Cleared samples were transferred to a fresh tube, mixed with 4x loading buffer containing l3>-mercaptoethanol, boiled for 10 min at 95 °C and analyzed by Western blotting. Samples were loaded into a SDS-PAGE gel and transferred to a 0.45 pm PVDF membrane (Roth). Membranes were blocked 1 h in 5% skim dry-milk in TBST (blocking buffer) and incubated with the corresponding antibodies diluted in 3% BSA-TBST overnight at 4 °C. After washing 3 times in TBST membranes were incubated in secondary HRP-conjugated antibodies (1 :5000 in blocking buffer) for 45 min at room temperature. Upon additional washing membranes were developed and imaged using FUSION software and imaging system.For immunoprecipitation (IP), samples were processed as above but lysed in IP buffer (Tris-HCI pH 7.4 50 mM, NaCI 150 mM, Igepal 0.5%50). Lysates were precleared in 30 pl Protein A / G PLUS Agarose beads (Santa Cruz; 1 :1 in IP buffer) for 1 h at 4 °C. Samples were incubated in the corresponding antibodies (1 pg antibody per mg of protein) overnight at 4 °C. 40 pl of Protein A / G PLUS Agarose beads were added for an additional hour and immunoprecipitates were washed three times in IP Buffer, resuspended in loading buffer and boiled for 5 min at 95 °C.Autophagy assaysAutophagy activity was measured using the following assays: (1 ) Western blotting analysis of p62 levels and LC3B-I / II ratio. (2) Quantification of the number ofautophagosomes by GFP-LC3 puncta formation. Cells were seeded in a 6-well plate and transfected the next day with the pBABE-GFP-LC3 plasmid. 24 h later cells were split into a black imaging 96-well plate (Eppendorf). After treatment cells were fixed in 4% PFA in PBS. Microscopic quantification was performed in the presence or absence of 100 nM Bafilomycin A1. (3) Luminescence analysis of the HiBiT-LC3 reporter. UMRC-6-HiBiT-LC3 cells were seeded at 4,000 cells / well in triplicate in a white 96-well plate (Santa Cruz). 24 h after plating, media was changed containing the desired treatment and incubated for the indicated periods of time. Nano-Gio HiBiT Lysis buffer containing LgBiT protein and substrate was added (1 :1 ), incubated in rotation for 10 min at room temperature and luminescence was measured in a plate reader. Parallel plates were set for assessment of the number of cells through Hoechst staining after fixation of the cells in 4% PFA-PBS.Semi-automatic drug screening in cell linesCompounds were dissolved in DMSO to create a concentration of 10 mM and were dispensed into white-bottom sterile polystyrene 384-well plates (Coming, 3570) at nine concentrations ranging from 5 nM to 50 pM in triplicate, using a Tecan D300e Digital Dispenser (Tecan, Switzerland I HP Inc., CA, USA) and D300e Control software. Negative controls with only DMSO were also included. To ensure capturing the linear growth phase of cells during drug treatment, cell number and assay period were optimized by seeding each cell line in triplicate at ten different cell densities, ranging from 100 to 3,000 cells per well, in 30 pl of cell culture medium in nine replicate assay plates, and performing a cell viability assay every 24 h for eight days. Cell proliferation was analyzed by normalizing luminescence values to day 0. For drug screening experiments, 500 cells per well were seeded in 30 pl of culture medium using a Multidrop Combi dispenser (Thermo Fisher Scientific, MA, USA) and assay plates were incubated at 37 °C and 5% CO2 for 4 days (96 h for bosutinib and saracatinib) or 5 days (120 h for dasatinib). The CellTiter-Glo assay (Promega) was performed to determine cell viability.Patient-derived tumor organoids (PDTOs) generation and cultureSurgical resections of RCC and UM primary tumors were collected at the University Hospital Essen following ethical approval by the Faculty of Medicine of the University of Duisburg-Essen (19-8553-BO and 21 -10013-BO) and written informed consent was obtained from all patients. Patient-derived tumor organoids (PDTOs) weregenerated from fresh primary tumors of RCC and UM, which were minced in very small pieces (1 -2 mm3) and dissociated in Dissociation Solution containing 1 mg / ml dispase (Life Technologies, 17105041 ) and 1 mg / ml collagenase la (Sigma-Aldrich, C9891 ) in RPMI for 15 min on ice and 50 pg / ml DNase I (Roche, 11284932001 ) for 15 min at 37 °C. Organoids were cultured and maintained in Growth Factor Reduced Matrigel (Coming, 356253) or Cultrex Reduced Growth Factor Basement Membrane Extract (Bio-Techne, 3536-005-02). A conditioned medium from L cells that express Wnt3a, R-spondin-3 and Noggin (L-WRN) was collected from L-WRN cells grown in 0.1 % FBS (but also possible up to 1 % or even 5% FBS instead of 10% FBS of the original conditioned medium) to prevent the differentiation and senescence of PDTOs and at the same time to prevent the apoptosis of L-WRN cells if grown in a serum- free medium, based on our previous studies in low serum conditions to inactivate mTORCI (Pe a-Llopis et al. EMBO J. 2011 ). The resulting final concentration of FBS in the organoid medium is below 0.025%, low enough to allow the almost indefinite successive passage of the PDTOs, while enough serum to prevent the apoptosis of L-WRN cells and allow the release of the Wnt3a, R-spondin-3 and Noggin.Drug screening in PDTOsDrug screens in PDTOs from ccRCC and UM were conducted in 384-well plates (Coming, 4588). PDTOs were treated with seven concentrations of dasatinib, seven concentrations of SW076956 and their combination in triplicate for 3 days (ccRCC PDTOs) or 4 days (UM PDTOs) and organoid viability was evaluated with CellTiter- Glo 3D. Two to three independent experiments were analyzed for synergistic effects.Protein extraction from PDTOsMedia was carefully removed by pipetting and PDTOs domes were washed in ice- cold PBS. PDTOs were incubated in Cell Recovery Media (Gibco) for 30 min on ice. Upon resuspension, PDTOs were transferred to Eppendorf tubes and spun at 1 ,200 g for three min at 4 °C. Pelleted PDTOs were washed in ice-cold PBS, pelleted, resuspended in 50 pl lysis buffer and processed for western blotting analysis as above.Reverse phase protein array (RPPA) data from ccRCC and UM patients from The Cancer Genome Atlas (TCGA) were downloaded from the Genomic Data Commons(GDC) data portal (https: / / portal.gdc.cancer.gov) with updated mutation and survival data for KIRC-TCGA and UVM-TCGA and analyzed as described in Pe a-Llopis et al. (2011 ).ChlP-Seq analysisRaw ChlP-Seq data were obtained from Gene Expression Omnibus (GEO) repository with accession numbers GSE4072359 and GSE6555560 and analyzed with Integrative Genomics Viewer (IGV) (Broad Institute, https: / / software.broadinstitute.org / software / igv) using mm9 genome.Quantitative Reverse-Transcribed PCR (qRT-PCR)RNA extraction and qRT-PCR analysis were performed. SRC mRNA expression was quantified using forward 5’-TGGCAAGATCACCAGACGG-3’ and reverse 5’- GGCACCTTTCGTGGTCTCAC-3’ primer pairs of SEQ ID NO: 22 and 23, respectively, and normalized to the housekeeping gene peptidylprolyl isomerase B (PPIB) using forward 5’-GAGGAAAGAGCATCTACGGTG-3’ and reverse 5’- GCTTCTCCACCTCGATCTTG-3’ primer pairs (SEQ ID NO: 26 and SEQ ID NO: 27, respectively).In Ovo Chick Chorioallantoic Membrane (CAM) AssayFreshly fertilized eggs were incubated at 38 °C with 50% atmospheric humidity in a humidified rotary incubator. On embryonic day 10, eggs were candled shining light into the eggshell to identify the chorioallantoic vein and a square was marked 1 cm away from the branching point. A little hole was made on the blunt end of the egg causing the air to escape allowing the “upper” chorioallantoic membrane to sag creating an artificial air chamber. This was followed by the opening of a 1 x1 cm window on the eggshell where the square was drawn. Immediately after opening the window, “the upper CAM” was lightly scuffed using a cotton tip. Then 50 pl of cell suspension (1.5 x 106cells in PBS) were graphted onto the CAM membrane. The window was then sealed with tape to avoid contamination or drying of the CAM and the eggs were kept at the incubator for another 7 days to allow tumor development. On day 17 post-fertilization eggs were sacrificed and the chicken was killed by decapitation after 15 min of ice incubation. Eggs were cut longitudinally and washed with PBS at least twice. Tumor identification was made following morphological (unified mass formation differentiated from other membrane components) andangiogenesis features (increased vasculature formed surrounding the tumor). Tumors were extracted, measured and cut in two for both histological (4% PFA fixation) and western blot analysis (flash frozen in liquid nitrogen).Statistical analysisGroups were compared and P values calculated using a two-tailed Student’s t test when equal variances or a Welch’s t test when unequal variances. Analysis of Variance (ANOVA) and Tukey post-hoc tests were calculated using IBM SPSS Statistics 29.0. Microsoft Excel and GraphPad Prism 9 and 10 were used to calculate the rest of statistical analyses.Example 1: BAP1 regulates SRC levels transcriptionally.Bioinformatics analysis on The Cancer Genome Atlas (TCGA) reverse-phase protein array (RPPA) data showed that one of the most upregulated proteins in BAP1 -mutant tumors from ccRCC patients is the non-receptor tyrosine kinase SRC, and it also showed a significant decrease in the phosphorylation levels of the inhibitory SRC site Y527 (Fig. 1 a). A similar analysis on the RPPA data from uveal melanoma (UM) tumors from TCGA (UVM-TCGA) also showed that tumors with BAP1 mutations have a decrease in SRC phosphorylation at Y527 (Fig. 1 b). These observations were confirmed by western blotting in a panel of BAP 1 -deficient cell lines from different entities reconstituted with an empty vector control or a vector encoding for an HA- tagged wild-type BAP1 (or a catalytically inactive p.C91 S mutant), including the renal cell carcinoma (RCC) cell line UMRC-6 (Fig. 1c), uveal melanoma cell line UPMM2 (Fig. 1 d), cholangiocarcinoma cell line TFK-1 (Fig. 1e) and breast carcinoma cell line HCC-1187 (Fig. 1f). In all cell lines tested a decrease in SRC levels was detected when cells expressed the wild-type form of BAP1 , but not when expressing the catalytically inactive form of BAP1 (p.C91 S), indicating that an active form of the epigenetic modulator BAP1 was required for regulating the SRC levels and therefore suggesting a transcriptional regulation.In addition, bioinformatics analysis of chromatin immunoprecipitation-sequencing (ChlP-Seq) data in bone marrow derived macrophages in mice indicates that BAP1 , along with Hcf-1 and Ogt (members of the Bap1 multiprotein complex to regulate gene expression), bind to the Src promoter in mice and might act as a negative regulator of Src expression. Furthermore, qRT-PCR data confirmed that these changes in SRClevels are transcriptionally regulated by BAP1 , since reconstitution with wild-type but not mutant BAP1 decreases SRC expression as determined in ccRCC cells UMRC-6 as well as in UM cells UPMM2 (Fig. 1 g,h). To rule out the possibility of an additional post-translational regulatory level, the stability of the SRC protein in UMRC-6 cells reconstituted with HA-BAP1 -WT or the p.C91 S mutant (or an empty vector control) was determined by blocking translation with cycloheximide over time. It was found that the half-life of SRC is similar in all cell lines tested. Taken together, these data indicate that BAP1 regulates SRC transcriptionally.Example 2: BAP1 -loss-mediated SRC activation promotes autophagy suppression.The inventors evaluated first the basal autophagy activity in UMRC-6 cells reconstituted with an empty vector, wild-type BAP1 or the p.C91 S mutant. Indeed, cells expressing the wild-type form of BAP1 had an elevated basal autophagy activity compared to the other cells, as determined by 1 ) lower levels of the autophagy substrate p62 by western blotting (Fig. 2a); 2) increased levels on the autophagosome-localized GFP-LC3 reporter detected by the GFP-LC3 puncta formation assay; the number of puncta is further increased with the addition of the lysosomal inhibitor Bafilomycin A1 (BafA1 ), indicating an activation of the autophagic flux rather than a block in autophagosomal maturation (Fig. 2b); and 3) decreased luminescence of the HiBiT-LC3 reporter, which ultimately measures the total LC3 levels (Fig. 2c). Taken together, these data indicate that UMRC-6 cells with loss of BAP1 have lower autophagy activity, which is activated upon reconstitution with wildtype BAP1 , but not the catalytically inactive p.C91 S BAP1 mutant. In addition, UMRC- 6 cells reconstituted with BAP1 mutants found in cancer patients indicate that only the wild-type BAP1 is able to induce autophagy (Fig. 2d), therefore suggesting that all clinically relevant BAP1 mutations analyzed behave as loss of function on their ability to modulate autophagy. Similar results were found in TFK-1 cholangiocarcinoma cells, UPMM2 uveal melanoma cells, HCC-1187 breast carcinoma cells, and ccRCC 786-0 cells with CRISPR-Cas9 wild-type BAP1 knocked-out using two different guided-RNAs.To ascertain whether the changes observed in autophagic flux indeed correlate with SRC activity, experiments modulating SRC activity were performed, both inhibiting (by siRNA knockdown or by treatment with the inhibitor of the SRC kinase dasatinib)or overexpressing SRC (using a V5-tagged SRC construct) in BAP7-deficient ccRCC UMRC-6 cells. SRC inhibition by dasatinib induced autophagic flux as determined by a decrease in p62 levels by western blotting (Fig. 2e), decreased luminescence of the HiBiT-LC3 reporter (Fig. 2f), increased GFP-LC3 puncta (Fig. 2g, h), and increased the activity of the catalytic subunit of the Class III-PI3KC1 , VPS34, as assessed by the number of GFP-FYVE puncta (Fig. 2i) in UMRC-6 cells treated with the inhibitor of the SRC kinase dasatinib. In addition, SRC depletion by siRNA also induced autophagic flux in UMRC-6 cells, as assessed by the HiBiT-LC3 reporter (Fig. 2j, k). Therefore, genetic or pharmacological inhibition of SRC induced autophagy in BAP1 - deficient cells through modulation of the VPS34 activity.Furthermore, overexpression of a V5-tagged SRC significantly increased the number of GFP-LC3 puncta per cell in UMRC-6-BAP1 cells, although these numbers did not increase upon addition of Bafilomycin A1 , suggesting an inhibition of the autophagosome-lysosomal fusion (Fig. 2I). By contrast, the empty vector control on the BAP1 reconstituted cells further increased the GFP-LC3 numbers with the addition of Bafilomycin A1.Taken together, these data strongly suggest that SRC is both necessary and sufficient to modulate autophagy activity in cells with BAP1 loss.Example 3: SRC binds to and phosphorylates BECN1.Autophagy is regulated through the Class III-PI3KC1 complex by multiple signals, including BECN1 phosphorylation by several kinases. To test whether SRC might regulate autophagy through binding and phosphorylation of BECN1 , UMRC-6 cells containing a shRNA targeting the endogenous SRC (or a non-targeting shRNA control) and a plasmid overexpressing a V5-tagged SRC (or empty vector, EV, control) were generated. SRC-V5 was found to co-immunoprecipitate with endogenous BECN1 (Fig. 3a).Furthermore, analysis of BECN1 -tyrosine phosphorylation also showed that endogenous BECN1 is phosphorylated in tyrosine residues in UMRC-6 cells (Fig. 3b), and that such phosphorylation decreases upon treatment with dasatinib, as determined by IP-WB. In addition, overexpression of SRC-V5 in both, UMRC-6-EV and UMRC-6-BAP1 cells increases BECN1 tyrosine phosphorylation, and suchphosphorylation goes away upon dasatinib treatment (Fig. 3c). BAP1 reconstitution in UMRC-6 cells increased the amount of VPS34 immunoprecipitated with BECN1 (Fig. 3c), in agreement with the data obtained from the GFP-2xFYVE assays (Fig. 2i) where more GFP dots are detected, and the binding VPS34 / BECN1 decreases when overexpressing SRC-V5. Taken together, these data suggest that BECN1 co- immunoprecipitate with SRC and that BECN1 -tyrosine phosphorylation in this context is regulated by SRC activity.To determine whether autophagic flux in UMRC-6 BAP 1 -deficient cells is dependent on the BECN1 tyrosine phosphorylation status, stable cell lines overexpressing Flag- BECN1 -WT, or two different BECN1 mutants containing non-phosphorylatable (BECN1 Y229 / 233 / 352F, hereafter referred as BECN-3F) or phospho-mimetic (BECN1 Y229 / 233 / 352E, hereafter referred as BECN-3E) mutations in the tyrosine residues involved in BECN1 dimerization and inactivation upon tyrosine phosphorylation were generated. Autophagic flux was assessed by measuring GFP- LC3 numbers in each cell line and showed that overexpression of BECN1 -WT induces autophagy, which was further induced by treatment with dasatinib. However, cells overexpressing the BECN1 mutants were not able to induce autophagy upon treatment with dasatinib (Fig. 3d,e), indicating that the above-mentioned residues in BECN1 are required for the autophagic response upon dasatinib treatment.To determine whether BECN1 phosphorylation affects the proliferation of BAP1 - deficient UMRC-6 cells, the stable cell lines expressing the unphosphorylatable (3F) or phosphomimetic (3E) forms of BECN1 were used. Cells overexpressing BECN1 - 3E (and therefore mimicking the status of a constitutively phosphorylated BECN1 ) were found to grow significantly faster than the ones stably expressing the wild-type of the 3F forms of BECN1 (or an empty vector control) (Fig. 3f). These data suggest that proliferation of BAP 1 -deficient ccRCC cell line UMRC-6 is dependent on the tyrosine phosphorylation status of BECN1 , suggesting a crucial role of BECN1 - dependent processes such as autophagy in this context.Example 4: SRC inhibition and autophagy induction are synergistic in vitro.Due to the autophagic flux modulation found in BAP 1 -deficient cells vs. reconstituted ones, it was tested whether inducing autophagy through one of the SRC inhibitors dasatinib, saracatinib or bosutinib or using an autophagy inducer that disrupts thebinding of BECN1 to its inhibitory partner BCL2 without inducing apoptosis (e.g. SW076956, SW063058 or Tat-Beclin-1 peptide) affects cell growth. First, the effects on autophagy of these compounds alone or in combination in UMRC-6 cells were tested. The results confirmed that SRC inhibitors and autophagy inducer both, indeed, induced autophagic flux as detected by the levels of p62 and lipidated LC3B by western blotting (Fig. 4a), HiBiT-LC3 luminescence (Fig. 4b, c) or autophagosome numbers using the GFP-LC3 reporter (Fig. 4d,e).Next, UMRC-6 cells were tested for growth in vitro when treated with combinations of dasatinib with SW076956 or of dasatinib with either Tat-Beclin-1 peptide or Tat- Beclin-1 -L11 S as well as with relevant controls. Unexpectedly, while both the SRC inhibitor dasatinib as well as the autophagy inducers SW076956 and Tat-Beclin-1 peptide inhibited cell growth independently, the magnitude of the effect was larger when treated with a combination of both, i.e. when treated with a combination product according to the present invention (Fig. 4f,g). In contrast, treatment with rapamycin, an mTORCI inhibitor, did not affect cell growth in this context (Fig. 4h). A high- throughput drug combination screen for dasatinib and SW076956 in UMRC-6 cells, testing 8 concentrations of each drug (and all possible combinations) and measuring the cell viability after 3 days of drug treatment was performed. Potential synergy / antagonism was analyzed using the Loewe model in Combenefit software. Surprisingly, the inventors observed a strong synergy of dasatinib and SW076956 with BAP1 -loss (EV and p.C91 S mutant), but not with wild-type BAP1 reconstitution (Fig. 4i,j). Similar results were observed in the high-throughput drug synergy screen of the cholangiocarcinoma cell line TFK-1 , with stronger synergism upon BAP1 loss (Fig. 4k, I). In analogous experiments, synergy / antagonism of the combination of:- other SRC inhibitors, namely saracatinib and bosutinib, with the autophagy inducer SW076956 (Fig. 5a-d); or- the SRC inhibitors dasatinib or saracatinib with another autophagy inducer, namely SW063058 (Fig. 6a and b), was also shown with respect to cell viability in the context of BAP1 loss in UMRC-6 and TFK-1 cells.Taken together, with these results the inventors show that SRC inhibitors (such as, for example, dasatinib, saracatinib or bosutinib) act synergistically with autophagyinducers (such as for example SW076956, SW063058 or Tat-Beclin-1 peptide) in the context of BAP 1 loss.Example 5: SRC inhibition and autophagy induction are synergistic in vitro.To ascertain the potential of using these compounds as treatment for BAP1 -deficient cancers, UMRC-6 cells were initially implanted subcutaneously in mice. Unfortunately, UMRC-6 cells were unable to grow in mice. Therefore, to strengthen the replacement, reduction and refinement of laboratory animals, UMRC-6 cells were implanted into the chorioallantoic membrane (CAM) of fertilized chick eggs. Cells were engrafted in PBS or in PBS containing dasatinib, SW076956 or a combination of both. Tumors were harvested 7 days later and, again, those treated with the combination of dasatinib and SW076956 showed a synergistic effect in that they were of significantly smaller size than the vehicle controls (Fig. 7a, b).To further confirm that autophagy contributes to regulate the growth of the CAM models, the inventors used Tat-BECN1 peptide (known to induce autophagy), which resulted in a dramatic decrease of tumor size. Tat-BECN1 peptide treatment in BAP1 - deficient UMRC-6 cells induced autophagy in vitro and decreased the tumor size in ovo when compared to the Tat-Beclin-1 -L11 S treatment (Fig. 7c, d).Example 6: SRC inhibition and autophagy induction are synergistic ex vivo in patient-derived tumor organoids (PDTOs).To further validate the synergistic effects of combinations in accordance with the present disclosure, i.e. of (a) an inhibitor of the tyrosine kinase SRC and (b) an autophagy inducer, to treat cancers, preferably cancers with BAP1 loss, the inventors used patient-derived tumor organoids (PDTOs) as preclinical models. After obtaining informed patient consent, PDTOs from fresh ccRCC and UM primary tumors were generated. Based on previous work with low serum to inhibit mTORCI (Pe a-Llopis et al. EMBO J. 2011 ), the inventors developed a cost-effective conditioned medium using low serum conditions to prevent the differentiation and senescence of PDTOs while preventing the apoptosis of L-WRN cells.A high-throughput drug combination screen for the inhibitor of the tyrosine kinase SRC dasatinib and the autophagy inducer SW076956 was performed in PDTOs from ccRCC and UM. It was found that in PDTOs with BAP1 loss the combination ofdasatinib and SW076956 displayed a strong synergy (Fig. 8a, c) whereas in PDTOs with wild-type BAP1 they did not (Fig. 8b, d).Overall, the inventors’ data indicates that: 1 ) BAP1 regulates SRC levels transcriptionally; 2) SRC binds and phosphorylates BECN1 ; 3) inhibitors of the tyrosine kinase SRC and autophagy inducers act synergistically in cancers, specifically in cancers with BAP1 loss, such that treatment with a combination product of the present disclosure decreases tumor growth. Taken together, but without wanting to be bound by theory, the surprising synergistic effect at the heart of the present disclosure, namely the synergistic effects of (a) an inhibitor of the tyrosine kinase SRC and (b) an autophagy inducer in cancer cells, appears to be the result of a mechanism where cancers, typically cancers with BAP1 loss, promote an upregulation of SRC to bind and phosphorylate BECN1 thereby suppressing autophagy (Fig. 8e). The inventors previously developed a reliable immunohistochemistry test to routinely identify tumors with BAP1 loss (Pe a-Llopis et al. Nat. Genet. 2012) for patient stratification in the clinic.The present disclosure now provides clinicians with an effective, previously unavailable treatment option for such patients, namely with a pharmaceutical combination product comprising (a) an inhibitor of the tyrosine kinase SRC and (b) an autophagy inducer.Many modifications and other embodiments of the invention set forth herein will come to mind to the one skilled in the art to which the invention pertains having the benefit of the teachings presented in the foregoing description and the associated drawings. Therefore, it is to be understood that the invention is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims

C l a i m s1. A pharmaceutical combination product comprising:(a) an inhibitor of the tyrosine kinase SRC; and(b) an autophagy inducer.

2. The pharmaceutical combination product according to claim 1 , wherein:(a) the inhibitor of the tyrosine kinase SRC is selected from the group consisting of:- SRC-specific miRNA inhibitors;- SRC-specific siRNA inhibitors;- SRC-specific proteolysis-targeting chimera (PROTAC);- dasatinib (A / -(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)- 1-piperazinyl]-2-methyl-4-pyrimidinyl]amino]-5-thiazole carboxamide), preferably in the form of monohydrate;- tirbanibulin (KX2-391 ; / V-Benzyl-2-(5-{4-[2-(4- morpholinyl)ethoxy]phenyl}-2-pyridinyl)acetamid);- bosutinib (SKI-606; 4-[(2,4-Dichlor-5-methoxyphenyl)amino]-6- methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]chinolin-3- carbonitrile);- ponatinib (AP24534; 3-(2-lmidazo[1 ,2-b]pyridazin-3-ylethynyl)-4- methyl- / V-(4-((4-methylpiperazin-1-yl)methyl)-3- (trifluoromethyl)phenyl)benzamide);- saracatinib (AZD0530; A / -(5-chloro-1 ,3-benzodioxol-4-yl)-7-[2-(4- methyl-1-piperazinyl)ethoxy]-5-[(tetrahydro-2 / - / -pyran-4-yl)oxy]- 4-quinazolinamine); and- eCF506 (CAS No. 1914078-41 -3; N-[4-[4-amino-1-[2-[4- (dimethylamino)-1-piperidinyl]ethyl]-1 H-pyrazolo[3,4-d]pyrimidin- 3-yl]-2-methoxyphenyl]-carbamic acid, 1 ,1 -dimethylethyl ester), or a pharmaceutically active salt or prodrug thereof; and(b) the autophagy inducer is selected from the group consisting of:- Beclin-1 -specific activators;- SW076956 (CAS No. 851717-83-4; N-(4-(5-(furan-2-yl)-1 -(4- methylbenzoyl)-4,5-dihydro-1 H-pyrazol-3- yl)phenyl)methanesulfonamide);- Tat-Beclin-1 ;- Tat-Beclin-1 peptide;- Tat-Beclin-1 -D11 peptide;- Tat-Beclin-1 -L11 peptide;- SW063058 (CAS No. 533876-30-1 ; 7-Bromo-5-(4-fluorophenyl)- 1 ,3,4,5-tetrahydro-4-(2-iodobenzoyl)-2H-1 ,4-benzodiazepin-2- one);- BRD1991 (CAS No. 2235468-02-5; 3,5-dichloro-N-((10-(1 - hydroxypropan-2-yl)-8, 12-dimethyl-11 -oxo-7,8,9, 10, 11 , 12- hexahydro-5H-benzo[9, 10][1 ]oxa[5]azacycloundecino[7,8- b]indol-7-yl)methyl)-N-methylbenzamide);- BRD5631 (CAS No. CAS 2446154-91 -0; N-[(3S,6S,7S)-7- methoxy-3,6,9-trimethyl-10-oxo-3,4,5,6,7,8,9,10-octahydro-2H-1 ,4,9-benzoxadiazacyclododecin-12 -y I] -[ 1 , 1 '-biphenyl]-4- carboxamide);- Tat-vFLIPa2; and- Tat-vFLIPa4, or a pharmaceutically active salt or prodrug thereof.

3. The pharmaceutical combination product according to claim 1 or claim 2, wherein:- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is Tat- Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is dasatinib, preferably in the form of monohydrate, and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is bosutinib and (b) the autophagy inducer is SW063058; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW076956; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is Tat-Beclin-1 ; or- (a) the inhibitor of the tyrosine kinase SRC is saracatinib and (b) the autophagy inducer is SW063058.

4. The pharmaceutical combination product according to any one of claims 1 to 3, comprising (a) the inhibitor of the tyrosine kinase SRC and (b) the autophagy inducer as separate entities.

5. The pharmaceutical combination product according to any one of claims 1 to 3, comprising (a) the inhibitor of the tyrosine kinase SRC and (b) the autophagy inducer in a single composition.

6. The pharmaceutical combination product according to any one of claims 1 to 5 for use in cancer treatment, wherein the cancer treatment targets BAP1- mutant or BAP1 -inactive cancer cells.

7. The pharmaceutical combination product for use according to claim 6, wherein the cancer treatment is treatment of- melanoma;- renal cancer;- pleural cancer; or- gallbladder cancer.

8. The pharmaceutical combination product for use according to claim 7, wherein the cancer treatment is treatment of- uveal melanoma (UM);- clear-cell renal cell carcinoma (ccRCC);- malignant pleural mesothelioma; orintrahepatic cholangiocarcinoma.

9. The pharmaceutical combination product for use according to any one of claims 6 to 8, wherein the treatment increases the cancer’s susceptibility to autophagy activation.

10. The pharmaceutical combination product for use according to any one of claims 6 to 9, wherein the treatment reduces the cancer’s growth, preferably the treatment reduces tumor size.

11. Use of (a) an inhibitor of the tyrosine kinase SRC; and (b) an autophagy inducer, for manipulating BAP1 -mutant or BAP1 -inactive cells.

12. The use according to claim 11 , wherein the BAP1 -mutant or BAP1 -inactive cells are manipulated such that Beclinl (BECN1) phosphorylation is decreased.

13. The use according to claim 11 or claim 12, wherein the BAP7-mutant or BAP1 - inactive cells are manipulated such that their proliferation is decreased.

14. The use according to any one of claims 11 to 13, wherein the BAP1 -mutant or BAP1 -inactive cells are manipulated in vitro.

15. The pharmaceutical combination product according to any one of claims 1 to 5 or the pharmaceutical combination product for use according to any one of claims 6 to 10 or the use according to any one of claims 11 to 14, wherein the autophagy inducer is not an inhibitor of the tyrosine kinase SRC.

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