Triple combination for use in the treatmet of cancer

Abstract

The present invention relates inter alia to pharmaceutical compositions and kits comprising (i) an inhibitor of the expression, activity and / or function of c-Raf, (ii) an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), or (iii) an inhibitor of the expression, activity and / or function of STAT3, for use in combination in the prevention and / or treatment of cancer.

Description

[0001] TRIPLE COMBINATION FOR USE IN THE TREATMET OF CANCER

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to a combined therapy against cancer which has proven to be particularly useful in the prevention and treatment of pancreatic cancer.

[0004] BACKGROUND

[0005] Pancreatic cancer is one of the most lethal cancer types, as it accounts for almost as many deaths as cases, and it has been projected as the second highest in mortality by 2030. With a 5-year survival rate of only 5%, the median survival is less than 6 months. However, minimal improvements have been made in the field of treatment. Currently, surgical resection remains the only curative option for early-stage local disease, which corresponds to 20-30% of patients. Unfortunately, due to late diagnosis, most patients cannot benefit from surgery, as at the time of diagnosis, they display advanced disease with metastasis. Cytotoxic chemotherapy is the standard treatment, with overall survival in the range of weeks to few months (Nevala- Plagemann et al., 2020). At present, no targeted therapeutic approaches are available for pancreatic ductal adenocarcinoma (PDAC) patients. So far, adjuvant treatment with 5- fluorouracil, leucovorin, irinotecan and oxaliplatin (FOLFIRINOX) has been demonstrated to provide the longest median overall survival (OS) (54 months) in patients with resectable disease (Conroy et al., 2018). For patients with advanced-stage or metastatic disease, the standard of care consists of modern combinations of highly cytotoxic agents such as nab- paclitaxel plus gemcitabine or FOLFIRINOX, but only provides modest improvements in OS in the range of weeks to months (Conroy et al., 2018; Von Hoff et al., 2013). Undoubtedly, there is urgent need to develop novel targeted therapies that directly block specific oncogenic pathways with reduced toxicity. Indeed, the recent consensus statement from the United States National Cancer Institute (NCI) indicated the need for targeted agents, predictive biomarkers, and improved preclinical models for PDAC (Philip et al., 2009).

[0006] Few preclinical studies have published combinations of EGFR or STAT3 inhibition with other compounds. In 2011 , Nagaraj et al. described that only the combination of dasatinib and erlotinib with gemcitabine overcame STAT3-mediated resistance of EGFR and Src inhibition both in vitro and in vivo. The pharmacological combination used in this study only includes the EGFR inhibitor Erlotinib (Nagaraj et al., 2011). In 2012, Navas et al. demonstrated that the inhibition of EGFR signaling with Erlotinib alone is not sufficient to target pancreatic cancer. Also, combination of phosphatidylinositol 3-kinase (PI3K) inhibitor with Stat3 shRNA (short hairpin RNA or small hairpin RNA (ribonucleic acid)) robustly inhibited proliferation of all tumor cell explants (Navas et al., 2012). In 2015, Zhou et al. demonstrated that STAT3 was significantly activated following MEK inhibition using AZD6244, PD98059 and Trametinib in Kras mutant pancreatic and colon cancer cells. As a result, dual inhibition of STAT3 and MEK exerts significant anti-tumor cell efficacy in Kras mutant pancreatic cancer cells in vitro (Zhao et al., 2015). In 2017, Sahu et al. disclosed that MAP2K (MEK) inhibitors were most effective in targeting PDAC spheroids, while combination with the multikinase inhibitor ponatinib was effective in targeting pancreatic cancer cells both in monolayer and spheroids. They also demonstrated that using xenograft models, combined treatment with a MEK inhibitor and ponatinib causes significant tumor regression. PDAC patient samples also provided evidence of increased STAT3 activation in PDAC tumors and MAPK1 (ERK) activation in liver metastases, implicating STAT3 and ERK as key drivers in primary tumors and metastases, respectively (Sahu et al., 2017). In 2019, Blasco et al. demonstrated that genetic ablation of Egfr and Rafi results in complete regression of a significant fraction of Kras / Trp53 driven tumors of small sizes. Yet, some small tumors and most tumors of bigger sizes remained refractory to this combined deletion (Blasco et al., 2019 and W02020020942 A1).

[0007] DESCRIPTION OF THE INVENTION

[0008] The inventors have shown that the simultaneous inhibition of the expression, activity and / or function of EGFR, RAF1 (also known as c-Raf) and STAT3 in PDAC results in a significant therapeutic effect with an extremely low toxicity. Previous studies with a double therapy against EGFR and RAF1 (also known as c-Raf) showed that this therapy was not effective in advanced and more aggressive PDACs of bigger sizes, which do not respond to the double combination. However, these advanced and more aggressive PDACs of bigger sizes can be effectively targeted with the triple combined therapy. The present invention represents an improved therapy in the treatment of cancer, particularly pancreatic cancer, since it leads to a complete tumor regression while its side effects are much smaller than other therapies described up to date.

[0009] The present invention relates inter alia to pharmaceutical compositions comprising (i) an inhibitor of the expression, activity and / or function of c-Raf, (ii) an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), or (iii) an inhibitor of the expression, activity and / or function of STAT3, for use in combination, e g. by means of simultaneous and / or sequential and / or separate administration, in the prevention and / or treatment of cancer.

[0010] Accordingly, the present invention provides a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

[0011] The present invention also provides a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

[0012] In addition, the present invention also provides a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

[0013] Moreover, the present invention provides a method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3. Suitably, the method of the present invention is a method for the prevention and / or treatment of pancreatic cancer, and for example is a method for the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma

[0014] Similarly, the present invention provides a method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

[0015] Also, the present invention provides a method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

[0016] In addition, the present invention provides a method for the prevention and / or treatment of cancer, in particular pancreatic cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a combination therapy according to the present invention.

[0017] Furthermore, the present invention provides the use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

[0018] In addition, the present invention provides the use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

[0019] Also, the present invention provides the use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

[0020] In addition, the present invention provides the use of a combination therapy according to the present invention in the manufacture of a medicament for the prevention and / or treatment of cancer, in particular pancreatic cancer.

[0021] The present invention provides a combination therapy comprising (i) an inhibitor of the expression, activity and / or function of c-Raf, (ii) an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (iii) an inhibitor of the expression, activity and / or function of STAT3, in particular wherein said combination therapy is for use in the treatment of cancer.

[0022] Furthermore, the present invention provides a combination therapy comprising (i) a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, (ii) a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (iii) a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, in particular wherein said combination therapy is for use in the treatment of cancer. In one embodiment, the combination therapy according to the present invention is formulated such that two or more of, for example two of, or for example three of, (i) the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, (ii) the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (iii) the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are co-administered. In one embodiment, the three components of the combination therapy are not co-formulated.

[0023] It will be understood that any of the below embodiments which relate to the pharmaceutical composition for use according to the present invention may equally relate to the method, uses, or combination therapies of the present invention.

[0024] As used herein, the terms ‘therapeutic effect’, ‘treatment’ or ‘treating’ include the control, mitigation, reduction or modulation of the state of the disease, disorder, or condition, or the symptoms of the disease, disorder, or condition.

[0025] As used herein, the terms ‘prophylactic effect’, ‘prophylaxis’, ‘prevention’, or ‘preventing’ refer to, in addition to preventing the occurrence of a disease, disorder, or condition, preventing symptoms of a disease, disorder, or condition in a subject, or preventing recurrence of symptoms of a disease, disorder, or condition, in an afflicted subject.

[0026] In one embodiment, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 are separate and distinct pharmaceutical compositions, i.e. three separate and distinct pharmaceutical compositions, which are administered simultaneously, and / or sequentially, and / or separately.

[0027] As used herein, the term ‘simultaneously’ (and the related term “simultaneous”) in the context of treatment means concurrent, i.e. at the same time, treatment with two or more different active agents or pharmaceutical compositions, wherein said different active agents or pharmaceutical compositions are implemented in the same phase, i.e. occur at the same time, but may vary with respect to conditions of administration (e.g. time period, route, dose, dosage form).

[0028] As used herein, the term ‘sequentially’ and the related term “sequential”) in the context of treatment means consecutive treatment with two or more different active agents or pharmaceutical compositions, typically directly adjacent in time, i.e. without a significant period without any treatment between the treatments with the two or more different active agents or pharmaceutical compositions.

[0029] As used herein, the term ‘separately’ (and the related term “separate”) in the context of treatment means treatment with two or more different active agents or pharmaceutical compositions at different periods of time. Typically, these periods are not directly adjacent in time, i.e. there is a time gap which is not insignificant between the treatments with the two or more different active agents or pharmaceutical compositions.

[0030] As used herein, the term “PROTAC” means a bifunctional molecule, comprising two active domains and a linker, typically where one of the active domains binds to an E3 ubiquitin ligase and the other active domain binds to a target protein. By recruiting the E3 ubiquitin ligase to the target protein, results in ubiquitination and subsequent degradation of the target protein.

[0031] It will be understood that when the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 are separate and distinct pharmaceutical compositions they may be administered in any and all combinations of simultaneous, sequential, and separate treatment. By way of example, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) may be administered simultaneously, followed by sequential or separate treatment with the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3. Alternatively, by way of example, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 may be administered, followed by sequential treatment with the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, followed by separate treatment with the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

[0032] In an alternative embodiment, two or more of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition, i.e. two or more of the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are formulated within a single pharmaceutical composition. Suitably, two of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition. For example, suitably, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) are the same pharmaceutical composition, i.e. the inhibitor of the expression, activity and / or function of c-Raf and the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) are formulated in the same pharmaceutical composition. When the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, refer to two separate and distinct pharmaceutical compositions, i.e. because two of them are the same pharmaceutical composition, said two separate and distinct pharmaceutical compositions may be administered in any and all combinations of simultaneous, sequential, and separate treatment.

[0033] Alternatively, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition.

[0034] For example, the present invention provides a pharmaceutical composition comprising: a. an inhibitor of the expression, activity and / or function of c-Raf; b. an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR); and c. an inhibitor of the expression, activity and / or function of STAT3.

[0035] When the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition, i.e. when the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are co-formulated, the pharmaceutical composition is suitably an aqueous solution composition, i.e. the inhibitors are coformulated in an aqueous vehicle, suitable for infusion or injection (e.g. the pharmaceutical composition is contained within one infusion bag).

[0036] Alternatively, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are not the same pharmaceutical composition, i.e. the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are not co-formulated, i.e. formulated within a single pharmaceutical composition.

[0037] Suitably, the inhibitor of the expression, activity and / or function of each one of c-Raf, EGFR or STAT3 comprises at least an inhibitor selected from: an inhibitor compound, an inhibitor antibody or an antigen binding fragment thereof, a peptide, a nucleotide sequence, a proteolysis targeting chimera (PROTAC), and any combination thereof. More suitably, the inhibitor of the expression, activity and / or function of each one of c-Raf, EGFR or STAT3 comprises an inhibitor compound, a PROTAC, or a combination thereof.

[0038] More suitably, when the inhibitor of the expression, activity and / or function of c-Raf, EGFR or STAT3 comprises a nucleotide sequence, the nucleotide sequence is or codifies for a guide RNA, an interfering RNA, such as a short hairpin RNA or small hairpin RNA (shRNA) or a micro RNA. shRNA molecules are well known in the art (see e.g. Navas et al., 2012) and typically comprise a double-stranded region of around 18-36 RNA nucleotides, wherein the strands are connected by a loop, which double-stranded region is complementary to a sequence present in a polynucleotide molecule, typically mRNA, encoding the target protein (e.g. c-Raf, EGFR or STAT3).

[0039] In a preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf does not inhibit the expression, activity and / or function of b-Raf. In a preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf does not inhibit c-Raf kinase activity. In a preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf inhibits the expression of c-Raf or promotes c-Raf degradation. In another preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf blocks c-Raf kinase independent activities.

[0040] Therefore, in one embodiment, the present invention relates to a pharmaceutical composition for use in the prevention and / or treatment of cancer, comprising an inhibitor of the expression, activity and / or function of c-Raf and an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), wherein the inhibitor of the expression, activity and / or function of c- Raf:

[0041] (i) does not inhibit c-Raf kinase activity; and / or

[0042] (ii) inhibits the expression of c-Raf; and / or

[0043] (iii) promotes c-Raf degradation; and / or

[0044] (iv) blocks c-Raf kinase independent activities.

[0045] Suitably, the inhibitor of the expression, activity and / or function of c-Raf is an inhibitor compound, and in particular is a small molecule inhibitor compound. As used herein, the term “small molecule” defines a molecule with a molecular weight of about 1200 g / mol or less, for example about 1000 g / mol or less, such as about 900 g / mol or less such as about 800 g / mol or less. Typically, a small molecule has a molecular weight of about 100 g / mol or more, for example about 200 g / mol or more, such as about 300 g / mol or more. Therefore, a small molecule may have a molecular weight of about 100-1200 g / mol, for example about 200-1000, for example about 300-800 g / mol.

[0046] Suitably, the inhibitor of the expression, activity and / or function of c-Raf is selected from the group consisting of sorafenib, vemurafenib, dabrafenib, regorafenib, cholecystokinin, fostamatinib, encorafenib, tovorafenib and LY3009120. For example, the inhibitor of the expression, activity and / or function of c-Raf is selected from the group consisting of regorafenib, cholecystokinin, fostamatinib, encorafenib, and tovorafenib. In an alternative embodiment, the inhibitor of the expression, activity and / or function of c-Raf is not sorafenib, vemurafenib, dabrafenib, regorafenib, cholecystokinin, fostamatinib, encorafenib, tovorafenib or LY3009120. For example, the inhibitor of the expression, activity and / or function of c-Raf is not regorafenib, cholecystokinin, fostamatinib, encorafenib, or tovorafenib. In one embodiment, the inhibitor of the expression, activity and / or function of c-Raf is an inhibitor of c-Raf kinase activity. In a preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf promotes c-Raf degradation. In a more preferred embodiment, the inhibitor of the expression, activity and / or function of c-Raf is a proteolysis targeting chimera (PROTAC).

[0047] The skilled person would be aware of suitable assays, which are known in the art, which are used to screen a putative inhibitor of the expression, activity and / or function of c-Raf for inhibitory activity. Typically, such an assay comprises a process of Western Blotting, and in particular may involve quantifying, via Western Blotting, the amount of c-Raf (i.e. for an inhibitor of expression), the amount of phosphorylated c-Raf (i.e. for an inhibitor of expression, activity, and / or function), or the amount of phosphorylated downstream signalling components, i.e. MEK and / or ERK (i.e. for an inhibitor of activity and / or function), in a cell line expressing c-Raf, for example inherently or following transfection. Exemplary assay protocols can be found in a number of literature documents including inter alia Awasthi et al. (2013), and Noeparast et al. (2018). Indeed, the present inventors have demonstrated inhibition of RAF1 (also known as c-Raf) expression in the examples of the present application (see Figures 1A and 1 B, Figures 2A, 2B, and 2C, Figure 6A and Figure 7A). Alternative assay formats, such as reporter gene assays, e.g. using luciferase or fluorescent tagged proteins, may also be suitable for screening a putative inhibitor of the expression, activity and / or function of c-Raf for inhibitory activity.

[0048] The half maximal inhibitory concentration (IC50) is a measure of the potency of an inhibitor. Specifically, IC50 is a quantitative measure that indicates how much of an inhibitor is needed to inhibit, in vitro, a given biological process or biological component, for example c-Raf activity. In one embodiment, the inhibitor of the expression, activity and / or function of c-Raf inhibits the expression, activity and / or function of each member of c-Raf with an IC50 value of less than about 50 pM, for example less than about 25 pM, for example less than about 10 pM, for example less than about 5 pM. More suitably, the inhibitor of the expression, activity and / or function of c-Raf inhibits the expression, activity and / or function of c-Raf with an IC50 value of less than about 1 pM (i.e. 1000 nM), for example less than about 0.75 pM (i.e. 750 nM), for example less than about 0.5 pM (i.e. 500 nM), for example less than about 0.25 pM (i.e. 250 nM). Most suitably, the inhibitor of the expression, activity and / or function of c-Raf inhibits the expression, activity and / or function of c-Raf with an IC50 value less than about 0.1 pM (i.e. 100 nM), for example less than about 0.05 pM (i.e. 50 nM), for example less than about 0.01 pM (i.e. 10 nM), for example less than about 0.001 pM (i.e. 1 nM). IC50 values may be determined by methods known by the skilled person.

[0049] Suitably, the inhibitor of the expression, activity and / or function of EGFR is an inhibitor compound, and in particular is a small molecule inhibitor compound.

[0050] Suitably, the inhibitor of the expression, activity and / or function of EGFR is selected from the group consisting of afatinib, abivertinib, alvocidib, canertinib, dacomitinib, erlotinib, gefitinib, brigatinib, icotinib, mobocertinib, neratinib, pelitinib, poziotinib, lapatinib, varlitinib, vandetanib, olmutinib, osimertinib, zanubrutinib, amivantamab, cetuximab, panitumumab, rindopepimut, necitumumab, nimotuzumab, zalutumumab, matuzumab, and combinations thereof. For example, the inhibitor of the expression, activity and / or function of EGFR is selected from the group consisting of abivertinib, alvocidib, canertinib, dacomitinib, mobocertinib, pelitinib, poziotinib, varlitinib, olmutinib, zanubrutinib, amivantamab, and rindopepimut.

[0051] The skilled person would be aware of suitable assays, which are known in the art, which are used to screen a putative inhibitor of the expression, activity and / or function of EGFR for inhibitory activity. Typically, such an assay comprises a process of Western Blotting, and in particular may involve quantifying, via Western Blotting, the amount of EGFR (i.e. for an inhibitor of expression), the amount of auto-phosphorylated EGFR (i.e. for an inhibitor of expression, activity, and / or function, or the amount of phosphorylated downstream signalling components, i.e. Ras and / or MEK and / or ERK (i.e. for an inhibitor of activity and / or function), in a cell line expressing EGFR, for example inherently or following transfection. Exemplary assay protocols can be found in a number of literature documents including inter alia Hirano et al. (2015) and Kancha et al. (2009). Indeed, the present inventors have demonstrated inhibition of EGFR expression in the examples of the present application (see Figures 1A and 1 B, and Figures 2A, 2B and 2C). Alternative assay formats, such as reporter gene assays, e.g. using luciferase or fluorescent tagged proteins, may also be suitable for screening a putative inhibitor of the expression, activity and / or function of EGFR for inhibitory activity.

[0052] In one embodiment, the inhibitor of the expression, activity and / or function of EGFR inhibits the expression, activity and / or function of EGFR with an IC50 value of less than about 50 pM, for example less than about 25 pM, for example less than about 10 pM, for example less than about 5 pM. More suitably, the inhibitor of the expression, activity and / or function of EGFR inhibits the expression, activity and / or function of EGFR with an IC50 value of less than about 1 pM (i.e. 1000 nM), for example less than about 0.75 pM (i.e. 750 nM), for example less than about 0.5 pM (i.e. 500 nM), for example less than about 0.25 pM (i.e. 250 nM). Most suitably, the inhibitor of the expression, activity and / or function of EGFR inhibits the expression, activity and / or function of EGFR with an IC50 value less than about 0.1 pM (i.e. 100 nM), for example less than about 0.05 pM (i.e. 50 nM), for example less than about 0.01 pM (i.e. 10 nM), for example less than about 0.001 pM (i.e. 1 nM).

[0053] Suitably, the inhibitor of the expression, activity and / or function of STAT3 is a shRNA or a PROTAC. In particular, the inhibitor of the expression, activity and / or function of STAT3 is a PROTAC.

[0054] Suitably, the inhibitor of the expression, activity and / or function of STAT3 is selected from lirodegimod (KT-333), SD-36 and SD-91 (see Zhou et al (2019) and Zhou et al (2021)), these molecules both being PROTACs. Thus, in one embodiment it is lirodegimod (KT-333). In one embodiment it is SD-36. In another embodiment it is SD-91 . Alternatively, the inhibitor of the expression, activity and / or function of STAT3 is selected from a variant of lirodegimod (KT- 333), a variant of SD-36 and a variant of SD-91 , which variants are also suitably PROTACs. For example, the inhibitor of the expression, activity and / or function of STAT3 is a PROTAC comprising the same E3 ubiquitin ligase binding moiety as lirodegimod (KT-333), SD-36, i.e. lenalidomide or pomalidomide, or SD-91 , i.e. lenalidomide, but a different STAT3 binding moiety to lirodegimod (KT-333), SD-36 or SD-91 (see Zhou et al (2019) and Zhou et al (2021). Alternatively, the inhibitor of the expression, activity and / or function of STAT3 is a PROTAC comprising a different E3 ubiquitin ligase binding moiety to lirodegimod (KT-333), SD-36 or SD-91 , but the same STAT3 binding moiety as lirodegimod (KT-333), SD-36, i.e. SI-109, or SD-91 , i.e. SI-191 , (see Zhou et al (2019) and Zhou et al (2021) The linker between the two binding moieties may be the linker used in lirodegimod (KT-333), SD-36 or SD-91 or may be a variant thereof. Further example inhibitor of the expression, activity and / or function of STAT3 which are PROTACs are described in US2023 / 084113A1 , US2023 / 083015A1 , US2023 / 120381A1 , US2023 / 133504A1, EP4121055 A1 , US2023 / 159573A1 , US2023 / 190938A1 , W02020 / 206424A1, WO2020 / 198435A1 , WO2021 / 188696A1,

[0055] WO2021 / 195481A1, WO2023 / 226950A1, US11485750B1 , and US11746120B2, (which documents are herein incorporated by reference in their entirety) .

[0056] The skilled person would be aware of suitable assays, which are known in the art, which are used to screen a putative inhibitor of the expression, activity and / or function of STAT3 for inhibitory activity. Typically, such an assay comprises a process of Western Blotting, and in particular may involve quantifying, via Western Blotting, the amount of STAT3 (i.e. for an inhibitor of expression), or the amount of phosphorylated STAT3 (i.e. for an inhibitor of expression, activity, and / or function), in a cell line expressing STAT3, for example inherently or following transfection. Exemplary assay protocols can be found in a number of literature documents including interalia Chen et al. (2021), Genini et al. (2017), and Zhou et al. (2019). See also the assays to determine STAT3 degradation described in US2023 / 084113 (herein incorporated by reference in its entirety), such as in Example 287 thereof. Indeed, the present inventors have demonstrated inhibition of STAT3 expression in the examples of the present application (see Figures 1A and 1 B, Figure 2A, and Figures 4A and 4B). Alternative assay formats, such as reporter gene assays, e.g. using luciferase or fluorescent tagged proteins, may also be suitable for screening a putative inhibitor of the expression, activity and / or function of EGFR for inhibitory activity.

[0057] In one embodiment, the inhibitor of the expression, activity and / or function of STAT3 inhibits the expression, activity and / or function of STAT3 with an IC50 value of less than about 50 pM, for example less than about 25 pM, for example less than about 10 pM, for example less than about 5 pM. More suitably, the inhibitor of the expression, activity and / or function of STAT3 inhibits the expression, activity and / or function of STAT3 with an IC50 value of less than about 1 pM (i.e. 1000 nM), for example less than about 0.75 pM (i.e. 750 nM), for example less than about 0.5 pM (i.e. 500 nM), for example less than about 0.25 pM (i.e. 250 nM). Most suitably, the inhibitor of the expression, activity and / or function of STAT3 inhibits the expression, activity and / or function of STAT3 with an IC50 value less than about 0.1 pM (i.e. 100 nM), for example less than about 0.05 pM (i.e. 50 nM), for example less than about 0.01 pM (i.e. 10 nM), for example less than about 0.001 pM (i.e. 1 nM).

[0058] In a preferred embodiment of the first aspect of the present invention, the composition comprises a therapeutically effective amount of an inhibitor of the expression, activity and / or function of c-Raf, a therapeutically effective amount of an inhibitor of the expression, activity and / or function of EGFR, a therapeutically effective amount of an inhibitor of the expression, activity and / or function of STAT3 and optionally a pharmaceutically acceptable excipient. Suitably, the present invention provides the pharmaceutical composition(s) for use according to the present invention, for use in the combination as described above, for use in the prevention and / or treatment of cancer, in particular pancreatic cancer. Similarly, the present invention provides any one of the combination therapies described above for use in the prevention and / or treatment of cancer, in particular pancreatic cancer. Suitably, the present invention relates to pharmaceutical composition(s) or combination therapies for use in the prevention and / or treatment of primary pancreatic cancer and / or the metastases derived from pancreatic cancer. For example, the present invention relates to the pharmaceutical composition(s) or combination therapies for use in the prevention and / or treatment of primary pancreatic cancer. Alternatively, the present invention relates to the pharmaceutical composition(s) or combination therapies for use in the prevention and / or treatment of metastases derived from pancreatic cancer, such as metastases in the liver, lung, lymph nodes, and peritoneum which are derived from pancreatic cancer. For example, the present invention relates to the pharmaceutical composition for use in the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma. Preferably, the pharmaceutical composition, for use in the combination as described above, is for use in tumor regression in a subject afflicted with said cancer.

[0059] Similarly, the present invention also provides the pharmaceutical composition, for use in the combination as described above, for use in tumor regression in a subject afflicted with pancreatic ductal adenocarcinoma (PDAC).

[0060] Moreover, the present invention provides the pharmaceutical composition for use according to the present invention, for use in the combination as described above, in the treatment of a subject who does not respond to treatment with a composition comprising an inhibitor of the expression, activity and / or function of c-Raf and / or treatment with a composition comprising an inhibitor of the expression, activity and / or function of EGFR.

[0061] Suitably, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 comprises the relevant active agent, i.e. the inhibitor of the activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3, and one or more pharmaceutically acceptable excipients, such as selected from the group consisting of adjuvants, antioxidants, bacteriostats, binders, carriers, detergents, diluents, disintegrants, emulsifiers, extenders, lubricants, salts, solvents, solutes, stabilisers, sugars, suspending agents, surfactants, such as non-ionic surfactants, oils, preservatives, thickeners, tonicity adjusting (osmotic) agents, buffers, viscosity enhancers, and water.

[0062] Suitably, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 comprises the relevant active agent, i.e. the inhibitor of the activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3, at a concentration which is required to achieve a prophylactic or therapeutic effect. As would be understood by the skilled person, the amount / concentration of active ingredient which is required to achieve a prophylactic or therapeutic effect will, of course, vary with the particular active ingredient, the route of administration, the subject under treatment or prophylaxis, including the type, species, age, weight, sex, and medical condition of the subject and the renal and hepatic function of the subject; and the particular disease, i.e. cancer, being treated or prevented, as well as its severity. An ordinarily skilled physician, veterinarian or clinician can readily determine and prescribe the effective amount of the active ingredient required to prevent, counter or arrest the progress of the cancer in question.

[0063] Suitably, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, may be suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous [bolus or infusion], and intraarticular), intranasal (also known as nasal administration), inhalation (including fine particle dusts or mists which may be generated by means of various types of metered dose pressurized aerosols, nebulizers or insufflators) insufflation, rectal, intraperitoneal, topical (including dermal, buccal, sublingual, and intraocular) or intrathecal administration. The most suitable route will depend on inter alia the subject under treatment or prophylaxis and the particular disease, i.e. cancer, being treated or prevented, as well as its severity. Particularly suitably, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, is independently suitable for oral or parenteral, in particular intravenous, administration. For example, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is independently administered via the oral or parenteral, in particular intravenous, administration route.

[0064] More suitably, each of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are administered via the same administration route, such as the oral or parenteral, in particular intravenous, administration route, but are not the same pharmaceutical compositions, i.e. the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are not co-formulated.

[0065] Alternatively, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are administered via two different, or via three different administration route, which suitably include the oral and / or the parenteral, in particular intravenous, administration route. For example, at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is administered via the oral administration route, and at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is not administered via the oral administration route. Alternatively, at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is administered via the parenteral, in particular intravenous, administration route, and at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is not administered via the parenteral, in particular intravenous, administration route.

[0066] Oral dosages of any one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, may suitably range between about 0.01 mg per kg of body weight per day (mg / kg / day) to about 100 mg / kg / day, suitably 0.01 mg per kg of body weight per day (mg / kg / day) to 10 mg / kg / day, and most suitably 0.1 to 5.0 mg / kg / day, for adult humans. Pharmaceutical compositions suitable for oral administration may be provided in the form of tablets or other forms of presentation provided in discrete units, such as capsules, cachets, pills, containing 0.01, 0.05, 0.1 , 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. Alternatively, pharmaceutical compositions suitable for oral administration may be presented as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid, for example as elixirs, tinctures, suspensions or syrups; or as an oil-in- water liquid emulsion or a water-in-oil liquid emulsion. A pharmaceutical composition suitable for oral administration typically contains from about 0.01 mg to about 500 mg of the active ingredient, i.e. the inhibitor of the activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), or the inhibitor of the expression, activity and / or function of STAT3, suitably from about 1 mg to about 100 mg of active ingredient.

[0067] Oral dosages of any one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the relevant active ingredient into association with one or more pharmaceutically acceptable carriers. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing into association the relevant active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired form. A tablet may be made by compression or moulding, optionally with one or more pharmaceutically acceptable carriers. Compressed tablets may be prepared by compressing, in a suitable machine, the relevant active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricating, surface active or dispersing agent. Moulded tablets may be made by moulding, in a suitable machine, a mixture of the powdered substance moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the relevant active ingredient therein. The pharmaceutical compositions therefore may be administered in a form suitable for immediate release or extended release. Immediate release or extended release can be achieved by the use of suitable pharmaceutical compositions comprising, or, particularly in the case of extended release, by the use of devices such as subcutaneous implants or osmotic pumps.

[0068] Pharmaceutical compositions for oral administration include suspensions which can contain, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents such as those known in the art; and immediate release tablets which can contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, calcium sulfate, sorbitol, glucose and / or lactose and / or other excipients, binders, extenders, disintegrants, diluents and lubricants such as those known in the art. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Disintegrators include without limitation starch, methylcellulose, agar, bentonite, xanthan gum and the like.

[0069] Moreover, any one of the pharmaceutical compositions comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, may be administered via the oral cavity by sublingual and / or buccal administration. Pharmaceutical compositions for topical administration in the mouth, for example buccally or sublingually, include lozenges comprising the relevant active ingredient in a flavoured basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as gelatin and glycerine or sucrose and acacia. Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene). Molded tablets, compressed tablets or freeze-dried tablets are exemplary forms which may be used. Suitable pharmaceutical compositions also include those formulating the relevant active ingredient, i.e. the inhibitor of the activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), or the inhibitor of the expression, activity and / or function of STAT3, with fast dissolving diluents such as mannitol, lactose, sucrose and / or cyclodextrins. Also included in such compositions may be high molecular weight excipients such as celluloses (Avicel) or polyethylene glycols (PEG). Such pharmaceutical compositions can also include an excipient to aid mucosal adhesion such as hydroxy propyl cellulose (HPC), hydroxy propyl methyl cellulose (HPMC), sodium carboxy methyl cellulose (SCMC), maleic anhydride copolymer (e.g., Gantrez), and agents to control release such as polyacrylic copolymer (e.g. Carbopol 934). Lubricants, glidants, flavors, coloring agents and stabilizers may also be added for ease of fabrication and use. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. For oral administration in liquid form, the active ingredient components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.

[0070] Pharmaceutical compositions of any one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. Pharmaceutical compositions suitable for parenteral administration, for example intravenous administration, may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example saline or water- for-injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described. Exemplary pharmaceutical compositions for parenteral administration include injectable solutions or suspensions which can contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1 ,3-butanediol, water, Ringer’s solution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid, or Cremaphor. Intravenously, the most suitable doses will range from about 0.1 to about 10 mg / kg / minute during a constant rate infusion. In one particular embodiment, the inhibitor of the expression, activity and / or function of c-Raf is administered orally. In another particular embodiment, the inhibitor of the expression, activity and / or function of EGFR is administered orally. In a further particular embodiment, the inhibitor of the expression, activity and / or function of STAT3 is administered parenterally, and in particular intravenously. In an alternative particular embodiment, the inhibitor of the expression, activity and / or function of STAT3 is administered orally. In one embodiment, each of the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGFR, and the inhibitor of the expression, activity and / or function of STAT3 is administered orally.

[0071] Suitably, the pharmaceutical compositions for use in the prevention and / or treatment of cancer, for use in the combination as described above, are useful for the prevention and / or treatment of cancer, such as pancreatic cancer condition in a mammal, in particular a human. Therefore, suitably, the pharmaceutical compositions for use according to the present invention are suitably administered to a mammal, in particular a human. Similarly, the combination therapies as described above, are useful for the prevention and / or treatment of cancer, such as pancreatic cancer, in a mammal, in particular a human. Therefore, suitably, the combination therapies for use according to the present invention are suitably administered to a mammal, in particular a human.

[0072] In one embodiment, the pharmaceutical compositions for use, combination therapies, and kits according to any aspect of the present invention are administered, for example in the context of being used in the prevention and / or treatment of cancer, e.g. pancreatic cancer, such that the inhibitor of the expression, activity and / or function of c-Raf is administered at a dose of 10 to 200 mg / day, such as 50 to 1800 mg / day, such as 100 to 1800 mg / day, such as 200 to 1500 mg / day, and / or such that the inhibitor of the expression, activity and / or function of EGFR is administered at a dose of 10 to 100 mg / day, such as 10 to 75 mg / day, such as 10 to 50 mg / day, such as 20 to 40 mg / day, and / or such that the inhibitor of the expression, activity and / or function of STAT3 is administered at a dose of 10 to 200 mg / day, such as 20 to 180 mg / day, such as 50 to 150 mg / day, such as 80 to 120 mg / day.

[0073] The present invention also provides a kit comprising (i) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, (ii) a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (iii) a third composition comprising an inhibitor of the expression, activity and / or function of STAT3. Furthermore, the present invention provides a kit comprising (i) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (ii) a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3. Alternatively, the present invention provides a kit comprising (i) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and an inhibitor of the expression, activity and / or function of STAT3, and (ii) second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR). Alternatively, the present invention provides a kit comprising (i) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and an inhibitor of the expression, activity and / or function of STAT3, and (ii) second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf.

[0074] Moreover, the present invention provides a kit comprising a single pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and an inhibitor of the expression, activity and / or function of STAT3.

[0075] Suitably, any of the above-described kits of the present invention further comprise instructions regarding use of the kit for the prevention and / or treatment of cancer, such as the prevention and / or treatment of pancreatic cancer, in particular the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma. Moreover, when any of the above-described kits of the present invention comprise two or more pharmaceutical compositions, the kits may further comprise instructions indicating that said pharmaceutical compositions are for administration simultaneously, and / or sequentially, and / or separately.

[0076] Accordingly, the kits of the present invention are suitably for use in the prevention and / or treatment of cancer, such as the prevention and / or treatment of pancreatic cancer, in particular the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma. Similarly, the present invention provides a method for the prevention and / or treatment of cancer, such as the prevention and / or treatment of pancreatic cancer, in particular the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of the contents of a kit according to the present invention. The kits of the present invention will comprise suitable packaging and / or materials that enable the use of the kit, i.e. administration of the pharmaceutical composition(s) packaged therein. For example, each pharmaceutical composition will typically be packaged in a separate container. Furthermore, by way of example, the kit may comprise a needle that facilitates parenteral, in particular intravenous, administration. Moreover, the kits may provide the pharmaceutical composition(s) packaged therein in unit dosage forms, bulk packages (e.g. multi-dose packages), or sub-unit doses. For example, the kits of the present invention may contain sufficient doses of the pharmaceutical composition(s) packed therein so as to enable effective administration of the pharmaceutical composition (s), and consequently to enable effective prevention and / or treatment of cancer, for an extended period of time, such as one week, two weeks, four weeks, six weeks, eight weeks, twelve weeks or more.

[0077] BRIEF DESCRIPTION OF THE FIGURES

[0078] Figure 1. A. Colony formation assay of Egfrlox / Iox; Raf1lox / IoxPDAC cell lines infected with Adeno-GFP and Adeno-CRE viral particles in combination with small-hairpin RNA (shRNA) against STAT3 and shControl (empty vector) as control. 5000 cells were seeded for 10 days. B. Western blot analysis of the expression of EGFR, RAF1 and STAT3 of cell-lysate extracts. GAPDH was used as loading control.

[0079] Figure 2. A. Colony formation assay of Egfrlox / Iox; Raf1lox / Iox; Stat3lox / IoxPDAC cell lines infected with Adeno-GFP and Adeno-CRE viral particles (R1-R5). 5000 cells were seeded for 10 days. B. Colony formation assay of Egfrlox / Iox; Raf1lox / Iox; Stat3lox / IoxPDAC cell lines (R1-R5 cell lines) infected with shRNA against EGFR and RAF1 (shRafl+shEgfr) and shControl (empty vector) as control. 5000 cells were seeded for 10 days. C. Western blot analysis of the expression of EGFR and c-RAF of cell-lysate extracts. GAPDH was used as loading control.

[0080] Figure 3. A. Tumor volume visualised by ultrasound imaging of tumors present in C57BL / 6 mice (n=3) orthotopically implanted with six representative RAF1 / EGFR Resistant KPeFC;Ra^iox / iox.^g^jox / iox.^a^iox / iox tumor ce|| |ines(R-J to R6) exposed to a tamoxifen (TMX) diet at the indicated times (arrow). Dotted line represents tumor sizes undetectable by ultrasound. B. Kaplan-Meier survival curve of tumor-bearing mice implanted with 12 independent KPeFC; Raf1l0X / l0X;Egf / j0X / l0X;Stat3l0X / l0Xtumor ceii lines either fed with a regular diet (n=36) or exposed to a tamoxifen (TMX) diet (n=36) for 100 days post-implantation. Mice were then fed with a regular diet for 200 additional days. Two TMX-diet mice died due to tumor-unrelated causes. The p value was obtained using unpaired T-test, p<0.005.

[0081] Figure 4. A. Colony formation assay of Egfrlox / Iox; Raf1lox / IoxPDAC cells infected with Adeno- GFP and Adeno-CRE viral particles in combination with treatment with 0.5 pM SD-36. 5000 cells were seeded for 10 days and culture medium was changed every day. B. H&E and IHC analysis of STAT3 expression in sections of two RAF1 / EGFR Resistant KPeFC; Raf1L / L; EgfrLILtumors treated with either vehicle or SD-36 (50 mg / kg). Scale bar represents 100 pm. C. Tumor volume visualised by ultrasound imaging of orthotopic tumors in C57BL / 6 mice (n=3) implanted with three independent KPeFC, Raf 1,',Egfr,;Stat3+,+tumor cell lines. Mice were treated with SD-36 (50 mg / kg, ip / qd) once their implanted tumors reached 40 to 50 mm3in size (arrows) for up to 60 days post-implantation. Dotted line represents tumor sizes undetectable by ultrasound. D. Kaplan-Meier survival curve of C57BL / 6 mice (n=12) implanted with four independent KPeFC; Raf1~l~ Egfr~,~;Stat3+,+tumor cell lines exposed to vehicle or to SD-36 for the indicated time. Animals were maintained for an additional 90 days without treatment. During this period of time none of the mice displayed detectable tumors as determined by ultrasound or died for unrelated causes. The p value was obtained using unpaired T-test, p<0.0001.

[0082] Figure 5. A. Colony formation assay of three representative KPeFC,Raf1~'~-,Egfr+l+;Statyl+tumor cell lines treated with afatinib (0.5 pM) and / or SD-36 (0.5 pM) for 10 days. B. Tumor volume visualised by ultrasound of tumors present in C57BL / 6 mice (n=3) implanted with four independent \<.PeFC Raf1~'~ Egfr+l+;Stat3+l+tumor cell lines and treated with afatinib (20 mg / kg, po, qd) and SD-36 (50 mg / kg, ip, qd) when the implanted tumors reached 60 to 100 mm3in size (arrows) for up to 60 days. Dotted line represents tumor sizes undetectable by ultrasound.

[0083] C. Kaplan-Meier survival curve of C57BL / 6 mice implanted with four independent KPeFC; / ?af7“ / _;Egfr+ / +;Sfat3+ / +tumor cell lines exposed to a combination of afatinib (20 mg / kg, po, qd) and SD-36 (50 mg / kg, ip, qd) (n=12) for the indicated period of time. Animals were kept for up to 100 days port-implantation without treatment. The p value was obtained using unpaired T-test, p<0.0001

[0084] Figure 6. A. Western blot analysis of c-Raf (i.e. Rafi) expression levels in whole cell extracts from PDX-derived cells either expressing control shRNAs (Ctrl) or shRNAs against Rafi (+). GAPDH was used as loading control. B. Proliferation of three independent PDX-derived cultures (PDX-DC1 to PDX-DC3) expressing shRNAs against Rafi and treated with either vehicle (Veh) or the combination of afatinib (0.5 pM) and SD-36 (0.5 pM) (Afat / SD36) at the indicated times (arrow). C. Tumor volume of three PDX-derived cultures (PDX-DC1 to PDX- DC3) expressing shRNAs against Rafi and orthotopically implanted in immunodeficient mice (n=3) and visualized by ultrasound. Mice were treated with a combination of afatinib (20 mg / kg, po, qd) and SD-36 (50 mg / kg, ip, qd) when they reached 50 to 120 mm3(arrows) until 60 days post-implantation. The dotted line indicates the limit of detection by ultrasound.

[0085] D. Representative images of the pancreas present in these mice. Pancreatic tumors present in the vehicle-treated mice (Vehicle) are indicated by T.

[0086] Figure 7. A. Western blot analysis of RAF1 expression levels in whole cell extracts from patient-derived organoids (PDO1 to PDO3) expressing an inducible shRNA against RAF1 in the absence (-) or presence (+) of doxycycline (Dox). GAPDH was used as loading control. B. Proliferation of three independent PDO cultures (PDO1 to PD03) expressing a Dox- inducible shRNA against RAF1 treated with either vehicle (solid circles) or a combination of DOX (1 pg / ml), afatinib (0.1 M) and SD-36 (1 pM) (Dox / Afat / SD36). The proliferation rate is represented by organoid area normalized to Day 0. Error bars indicate mean ± SEM. C. Tumor volume of two PDO cultures (PDO1 , PDO2) implanted subcutaneously in immunodeficient mice (n=3) and visualized by ultrasound. Mice were either treated with Vehicle (Veh) or with a combination of afatinib (20 mg / kg, po, qd), SD-36 (50 mg / kg, ip, qd) and doxycycline (2 mg / ml) (Dox / Afat / SD36) when tumors reached 100 to 200 mm3(arrows) for the indicated time. The dotted line indicates the limit of detection by ultrasound.

[0087] EXAMPLES

[0088] The present invention is exemplified and illustrated by the following examples, which are not intended to limit in any way the present invention:

[0089] Materials and Methods

[0090] Silencing of Egfr, Rafi and Stat3 was mediated with the lentiviral short hairpin RNA (shRNA) plasmids TRCN0000055218 (mouse), TRCN0000195646 (human), TRCN0000012628 (mouse) and TRCN0000071453 (mouse) respectively. Non-targeting shRNA vectors were used as control. The pharmacological treatments were conducted with the following agents: afatinib (20 mg / kg, daily), SD-36 (50 mg / kg, daily).

[0091] The triple elimination of EGFR, RAF1 and Stat3 eliminated all PDACs

[0092] Combined ablation of EGFR and RAF1 expression resulted in complete regression of a significant percentage (around 50%) of PDAC tumors (Sensitive, S, tumors) driven by Kras / Trp53 mutations in genetically engineered mice (Blasco et al., Cancer Cell 2019). The experiments were done in tumors where the oncogene Kras is mutated as well as the oncosupressor gene p53 (For short KPeFC in figures). Basically, the cases of pancreatic ductal adenocarcinoma, both in human and in mouse models, have two major drivers: Oncogenic Kras mutations and p53 mutations. In the study of Blasco et al., 50% of the mouse PDACs (Resistant, R, tumors) were insensitive to this therapeutic strategy. Surprisingly, the triple elimination of EGFR, RAF1 and Stat3 eliminated all PDACs: S and R tumors.

[0093] STAT3 inhibition (genetic or pharmacological) resulted in cell death of R tumor cells The expression of STAT3 was targeted by shRNAs after genetic deletion of Egfr and Rafi in R cells by Adeno-GFP and Adeno-GFP-CRE infection. Importantly, this combined therapeutic strategy resulted in cell death of R tumor cells (Figure 1A and 1 B). Notably, downregulation of Stat3 did not affect R cells that express Egfr and Rafi (Figure 1A and 1B). These results were further validated by a pharmacological approach, using the STAT3 PROTAC degrader SD-36 (Figure 4A). R tumor cells, previously infected with Adeno-GFP and Adeno-GFP-CRE virus to eliminate EGFR and the Rafi (floxed targets), were treated with SD-36 for 10 days in a colony formation assay (Figure 4A). Elimination of the targets was very efficient. SD-36 effectively degraded STAT3 and has no toxic side-effects, as R cells that express Egfr and Rafi grew similarly as the vehicle control (Figure 4A).

[0094] Triple deletion in vitro in resistant (R) cells

[0095] A genetic approach was also performed: specifically, triple elimination of Egfr, Rafi and Stat3. For this, we generated a PDAC mouse model that carries the three floxed alleles: Egfrlox / Iox; Raf1lox / Iox; Stat3lox / Iox. 30 PDAC cell lines were generated for in vitro studies. The simultaneous elimination of the three floxed targets by Adeno-CRE infection resulted in complete cell death of the 30 cell lines. Figure 2A shows an exemplary 5 cell lines. Interestingly, the few small colonies that remained were mostly apoptotic and could not be collected for further studies. We identified these cell lines as R based on their response to the elimination of EGFR and Rafi by shRNAs (Figure 2B and 2C). From the 30 cell lines, 12 were R. Therefore, elimination of the three targets efficiently induced cell death in R cells.

[0096] Triple deletion in vivo

[0097] We also studied the potential therapeutic value of the simultaneous elimination of Egfr, Rafi and Stat3 in vivo. We performed orthotopic studies, by inoculating three KPeFC; Egfrlox / Iox; Rafl'ox / iox; Stat3lox / I°xtumor cell lines in the pancreas of immunocompetent mice (C57BL / 6) mice. These cell lines were R to the elimination of Egfr and Rafi (Figure 4B and 4C) and carry the Rosa26CreERT2 allele. Upon exposure to tamoxifen, CreERT2 recombinase was activated resulting in the elimination of the floxed alleles in the tumor cells. For a therapeutic approach, we performed an orthotopic therapeutic study, in which the tamoxifen diet was provided to tumor bearing mice. Strikingly, we observed regression of tumors of different sizes (10-100 mm3) after a treatment period of approximately three weeks. Tumor monitoring was continued for additional 35 days under tamoxifen treatment to confirm that triple elimination in the tumoral cells induced complete and sustained tumor regression (Figure 3A). Therefore, the simultaneous elimination of the three targets in vivo produced complete and irreversible tumor regression. No tumor resistance was observed until 300 days after implantation (Figure 3B). Upon treatment with tamoxifen, the expression of the gene Rafi was eliminated genetically. We also included in the combined treatment afatinib (a tyrosine receptor kinase inhibitor against EGFR) and SD-36 (PROTAC degrader against STAT3). First, treatment of mice, with PDAC tumors, with SD-36 resulted in effective degradation of STAT3 (Figure 4B). Next, in vivo experiments of implanted tumors (20-50 mm3) that did not express Egfrand Rafi resulted in complete and irreversible tumor regression, without any development of resistance (Figure 4C and 4D). Further combination of SD-36 with afatinib in PDAC cells that lack expression of c- Raf, showed no proliferation of tumor cells in vitro (Figure 5A). Also, in vivo treatment with afatinib and SD-36 of mice with Raft1' tumors of a large size (60-100 mm3) resulted in complete and long-lasting regression of these tumors (Figure 5B and 5C). On the contrary, RafT / _tumors treated only with afatinib did not regress and mice had to be sacrificed within three weeks after implantation (Figure 5C). To further expand the clinical data, we silenced, using shRNAs, the expression of c-Raf in human PDAC cells derived from patient-derived xenografts (PDX) (Figure 6A). Next, we treated, in vitro, three PDX-derived cell lines (PDX-DC1 to PDX-DC3) with the combination of afatinib and SD-36, resulting in efficient cell death (Figure 6B). A similar in vivo approach involving implanted tumors of a large size (40-100 mm3) resulted in regression of PDACs, while vehicle-treated tumors grew rapidly. Equally importantly, 3D cultures from fresh human tumors, indicated as patient-derived organoids (PDOs) were infected with a vector expressing shRNA against Rafi inducible by doxycycline (Dox). Thus, treatment of these cultures with Dox, resulted in effective Rafi silencing (Figure 7A). Moreover, combined treatment with Dox, afatinib and SD-36 resulted in complete cell death of PDOs (Figure 7B). Finally, in vivo experiments with implanted PDOs in immunodeficient mice, validated previous findings, in particular since treatment with Dox, afatinib and SD-36 in combination led to regression of subcutaneous tumors of approx. 100 mm3, compared to vehicle treated tumors (Figure 7C). These data validate the efficacy of the combined therapy in human-derived tumors.

[0098] REFERENCES

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Claims

CLAIMS:

1. A pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitorof the expression, activity and / or function of EGF Receptor (EGFR) and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

2. A pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

3. A pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 for use in the prevention and / or treatment of cancer, for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and for use in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

4. The pharmaceutical composition for use according to any one of claims 1 to 3, wherein the inhibitor of the expression, activity and / or function of each one of c-Raf, EGFR and STAT3 comprises at least an inhibitor selected from: an inhibitor compound, an inhibitor antibody or an antigen binding fragment thereof, a peptide, a nucleotide sequence, a proteolysis targeting chimera (PROTAC), and any combination thereof.

5. The pharmaceutical composition for use according to claim 4, wherein the inhibitor of the expression, activity and / or function of each one of c-Raf, EGFR and STAT3 comprises at least an inhibitor selected from an inhibitor compound, a proteolysis targeting chimera (PROTAC), and a combination thereof.

6. The pharmaceutical composition for use according to any one of claims 1 to 5, wherein the inhibitor of the expression, activity and / or function of c-Raf does not inhibit the expression, activity and / or function of b-Raf; and / or wherein the inhibitor of the expression, activity and / or function of c-Raf does not inhibit c-Raf kinase activity.

7. The pharmaceutical composition for use according to any one of claims 1 to 6, wherein the inhibitor of the expression, activity and / or function of c-Raf is selected from the group consisting of sorafenib, vemurafenib, dabrafenib, regorafenib, cholecystokinin, fostamatinib, encorafenib, tovorafenib and LY3009120.

8. The pharmaceutical composition for use according to any one of claims 1 to 6, wherein the inhibitor of the expression, activity and / or function of c-Raf is not sorafenib, vemurafenib, dabrafenib, regorafenib, cholecystokinin, fostamatinib, encorafenib, tovorafenib or LY3009120.

9. The pharmaceutical composition for use according to any one of claims 1 to 6, wherein the inhibitor of the expression, activity and / or function of c-Raf is a proteolysis targeting chimera (PROTAC).

10. The pharmaceutical composition for use according to any one of claims 1 to 9, wherein the inhibitor of the expression, activity and / or function of EGFR is selected from the group consisting of afatinib, abivertinib, alvocidib, canertinib, dacomitinib, erlotinib, gefitinib, brigatinib, icotinib, mobocertinib, neratinib, pelitinib, poziotinib, lapatinib, varlitinib, vandetanib, olmutinib, osimertinib, zanubrutinib, amivantamab, cetuximab, panitumumab, rindopepimut, necitumumab, nimotuzumab, zalutumumab, matuzumab, and combinations thereof.

11. The pharmaceutical composition for use according to any one of claims 1 to 10, wherein the inhibitor of the expression, activity and / or function of STAT3 is a shRNA or a proteolysis targeting chimera (PROTAC).

12. The pharmaceutical composition for use according to claim 11, wherein the inhibitor of the expression, activity and / or function of STAT3 is a proteolysis targeting chimera (PROTAC).

13. The pharmaceutical composition for use according to claim 12, wherein the inhibitor of the expression, activity and / or function of STAT3 is selected from lirodegimod (KT-333), SD-36 and SD-91.

14. The pharmaceutical composition for use according to claim 12, wherein the inhibitor of the expression, activity and / or function of STAT3 is selected from a variant of lirodegimod (KT- 333), a variant of SD-36 and a variant of SD-91.

15. The pharmaceutical composition for use according to any one of claims 1 to 14, whereinthe pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 are separate and distinct pharmaceutical compositions which are administered simultaneously, and / or sequentially, and / or separately.

16. The pharmaceutical composition for use according to any one of claims 1 to 14, wherein two of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition, i.e. two of the inhibitor of the expression, activity and / or function of c-Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are formulated within a single pharmaceutical composition.

17. The pharmaceutical composition for use according to any one of claims 1 to 14, wherein the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are the same pharmaceutical composition, i.e. the inhibitor of the expression, activity and / or function of c- Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are formulated within a single pharmaceutical composition.

18. The pharmaceutical composition for use according to any one of claims 1 to 14, wherein the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are not the same pharmaceutical composition, i.e. the inhibitor of the expression, activity and / or function of c- Raf, the inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and the inhibitor of the expression, activity and / or function of STAT3 are formulated within a single pharmaceutical composition.

19. The pharmaceutical composition for use according to any one of claims 1 to 18, for use in the prevention and / or treatment of pancreatic cancer.

20. The pharmaceutical composition for use according to any one of claims 1 to 19, for use in the prevention and / or treatment of pancreatic intraepithelial neoplasia and / or of pancreatic ductal adenocarcinoma.

21. The pharmaceutical composition for use according to any one of claims 1 to 20, for use in tumour regression in a subject afflicted with cancer.

22. The pharmaceutical composition for use according to claim 21 , for use in tumor regression in a subject afflicted with pancreatic ductal adenocarcinoma (PDAC).

23. The pharmaceutical composition for use according to any one of claims 1 to 22, in the treatment of cancer in a subject who does not respond to treatment with a composition comprising an inhibitor of the expression, activity and / or function of c-Raf and / or treatment with a composition comprising an inhibitor of the expression, activity and / or function of EGFR.

24. The pharmaceutical composition for use according to any one of claims 1 to 23, wherein the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are each independently suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, and intraarticular), intranasal, inhalation, insufflation, rectal, intraperitoneal, topical (including dermal, buccal, sublingual, and intraocular) or intrathecal administration.

25. The pharmaceutical composition for use according to claim 24, wherein the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, are each independently suitable for oral or parenteral, in particular intravenous, administration26. The pharmaceutical composition for use according to claim 24 or claim 25, wherein at least one of the pharmaceutical composition comprising an inhibitor of the expression,activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is administered via the oral administration route, and at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is not administered via the oral administration route.

27. The pharmaceutical composition for use according to claim 24 or claim 25, wherein at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is administered via the parenteral, in particular intravenous, administration route, and at least one of the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and the pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 is not administered via the parenteral, in particular intravenous, administration route.

28. The pharmaceutical composition for use according to any one of claims 1 to 27, for use in the prevention and / or treatment of cancer in a mammal, in particular a human.

29. A method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

30. A method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), in combination with a pharmaceutical compositioncomprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.31 . A method for the prevention and / or treatment of cancer, comprising administering to a subject in need thereof a prophylactically or therapeutically effective amount of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

32. Use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

33. Use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3.

34. Use of a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 in the manufacture of a medicament for the prevention and / or treatment of cancer, in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, and in combination with a pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

35. A kit comprising (i) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf, (ii) a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR), and (iii) a third composition comprising an inhibitor of the expression, activity and / or functionof STAT3.

36. A kit comprising:(a) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and an inhibitor of the expression, activity and / or function of STAT3; or(b) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR) and a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and an inhibitor of the expression, activity and / or function of STAT3; or(c) a first pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of STAT3 and a second pharmaceutical composition comprising an inhibitor of the expression, activity and / or function of c-Raf and an inhibitor of the expression, activity and / or function of EGF Receptor (EGFR).

37. A kit according to claim 35 or claim 36, further comprising instructions regarding use of the kit for the prevention and / or treatment of cancer, such as the prevention and / or treatment of pancreatic cancer.

38. A kit according to any one of claims 35 to 37, further comprising instructions indicating that the two or more pharmaceutical compositions of the kit are for administration simultaneously, and / or sequentially, and / or separately.

39. A kit according to any one of claims 35 to 38, for use in the prevention and / or treatment of cancer, such as the prevention and / or treatment of pancreatic cancer.

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