Il-15 in combination with chemotherapy for treating cancer

Abstract

The present invention relates to a pharmaceutical composition comprising IL-15 or a derivative thereof, for use in combination with a chemotherapeutic agent in the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent, and associated methods. The present invention also relates to methods of sensitising cells to chemotherapeutics and enhancing efficacy of chemotherapeutics.

Description

[0001] IL-15 IN COMBINATION WITH CHEMOTHERAPY FOR TREATING CANCER

[0002] Field of Invention

[0003] The present invention relates to the treatment of cancer using a combination of IL-15 or a derivative thereof and a chemotherapeutic agent.

[0004] Background

[0005] Chemotherapeutics remain standard of care for many cancer patients, include high grade serous ovarian cancer following metastasis to the peritoneal cavity. It is becoming clear that chemotherapy can initiate adaptive immune responses to the cancer, however, it remains unclear to what extend innate Natural Killer (NK) cells contribute. Previous transcriptomic analysis of human ovarian cancer patient data suggests that NK cells are enriched after chemotherapy.

[0006] There is a need to develop further therapies that can enhance the efficacy of chemotherapeutic treatments.

[0007] Summary of the Invention

[0008] The present invention provides a therapy for the treatment of cancer. The present inventors have surprisingly shown herein that Interleukin-15 (IL-15) treatment can be used to expand NK cells and / or CD8+ T cells and this treatment can be used to enhance the efficacy of subsequent chemotherapeutic treatment.

[0009] An aspect of the invention relates to a pharmaceutical composition comprising IL-15 or a derivative thereof, for use in combination with a chemotherapeutic agent in the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent.

[0010] An aspect of the invention relates to a method of treating cancer in a subject, comprising administering a pharmaceutical composition comprising IL-15 or a derivative thereof, in combination with a chemotherapeutic agent to said subject, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the platinum-based chemotherapeutic agent.

[0011] An aspect of the invention relates to the use of a pharmaceutical composition comprising IL-15 or a derivative thereof, for the manufacture of a medicament for the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to a chemotherapeutic agent.

[0012] An aspect of the invention relates to an in vitro, ex vivo or in vivo method of sensitising cells to a chemotherapeutic agent comprising: contacting a population of cells to IL-15 or a derivative thereof, contacting said population of cells to a chemotherapeutic agent.

[0013] An aspect of the invention relates to an in vitro, ex vivo or in vivo method of enhancing efficacy of a chemotherapeutic agent comprising: exposing a population of cells to IL-15 or a derivative thereof, exposing said population of cells to a chemotherapeutic agent.

[0014] An aspect of the invention relates to a method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for therapy.

[0015] An aspect of the invention relates to a method of therapy monitoring comprising: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, comparing the level of NK cells and / or CD8+ T cells with a reference value, determining the efficacy of the therapy based on the level of NK cells and / or CD8+ T cells compared to said reference value, wherein the subject has received, is currently receiving or will receive a therapy comprising IL- 15 or a derivative thereof and a chemotherapeutic agent, wherein the IL-15 is administered prior to said chemotherapeutic agent.

[0016] An aspect of the invention relates to a method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises identifying the subject as having one of more of the following; increased expression, amplification of expression and / or mutation in the KRAS gene, reduced expression, loss of expression and / or a mutation in the p53 gene, reduced expression, loss of expression and / or a mutation in a Brea gene, and selecting said subject for therapy.

[0017] An aspect of the invention relates to a kit for the treatment of cancer comprising IL-15 or a derivative thereof and a chemotherapeutic agent.

[0018] Figures

[0019] Figure 1. IL-15 / IL-15Ra increases lymphocyte numbers and function. A) Schematic of IL- 15 / IL-15Ra dosing regime in tumour naive C57bl / 6 mice. B) Total immune and lymphocyte subset cell count in omental tissue in untreated and 11-15 / IL-15Ra treated mice. C) Representative NK1.1+NK cell population Ki67 gating. D) Percent of Ki67+ NK cells. E) Representative CD8+ T cell Ki67 gating. F) Percent of Ki67+ CD8+ T cells. G) NK subset cell number. H) Total immune and lymphocyte subset total cell count in the spleen in untreated and 11-15 / IL-15Ra treated mice. I) NK subset cell number. J) Percent of Ki67+ NK cells. K) Percent of Ki67+ CD8+ T cells. L) Percentage of IFNg+ and GzmB+ NK cells with and without restim. M) Percentage of IFNg+ and GzmB+ CD8+ T cells with and without restim. Bars represent mean. Error bars represent SEM. T-tests (b,d,f,h,j-m) and two-way ANOVA (g, i) statistical tests were performed. Data are representative of 6 mice across 2 experiments, excluding intracellular staining of Ki67, IFNg and GzmB which is only representative of 1 experiment, p values - ns = nonsignificant, * = <0.05, ** = <0.01 , ***<0.001 , ****<0.0001 .Figure 2. IL-15 / IL-15Ra pretreatment improves efficacy of carboplatin in UPK10 model. A) Schematic of 11-15 / IL-15Ra and carboplatin dosing regime in UPK10 tumour bearing C57bl / 6 mice. B) Omentum tumour weight at day 24 endpoint. C) Total number of CD45+ cells in tumour. D) Total number of B cells, E) CD4+ T cells, F) CD8+ T cells and G) NK cells in the tumour. Error bars represent SEM. Oneway ANOVA statistical test was performed. n=15 mice, 3 independent experiments (b) and n=10 mice, 2 independent experiments (c-g). p values - ns = nonsignificant, * = <0.05, ** = <0.01 , ***<0.001 , ****<0.0001.

[0020] Figure 3. A single dose of 11-15SA results in a temporal expansion of lymphocytes, peaking 3 days post injection. A) IL-15 / IL-15Ra time course in tumour naive C57bl / 6 mice. One dose of IL-15SA was given 14, 7, 5, 3, or 1 day before takedown. PBS was given to the control group 1 day before takedown. B) Total immune and lymphocyte subset total cell count in omental tissue. C) Histogram representing Ki67 expression in NK cells and CD8 T cells (LHS) across timepoints. Quantification of Ki67+ NK and CD8 T cells (RHS). D) Proportion of NK cell maturation subsets across timepoints. E) Total immune and lymphocyte subset total cell count in spleen tissue. F) Percent of Granzyme B positive (LHS) and IFNy positive (RHS) NK cells. G) Percent of Granzyme B positive (LHS) and IFNy positive (RHS) CD8 T cells. H) Number of CD8 memory T cells. I) NK cell and CD8 T cell count in blood. Error bars represent SEM. Statistical tests are one-way anova. n = 9 mice per group, 3 independent experiments, p values - ns = nonsignificant, * = <0.05, ** = <0.01 , ***<0.001 , ****<0.0001 .

[0021] Figure 4. IL-15 / IL-15Ra pre-treatment improves efficacy of carboplatin in UPK10 model. A) Schematic of 11-15 / IL-15Ra and carboplatin dosing regime in UPK10 tumour bearing C57bl / 6 mice. B) Omentum tumour weight at day 24 endpoint. C) Probability of survival. D) Total number of CD45+ cells, B cells, CD4+ T cells, CD8+ T cells and NK cells in the tumour. Error bars represent SEM. One-way ANOVA statistical test was performed. n=15 mice, 3 independent experiments (b) and n=10 mice, 2 independent experiments (c-g). p values - ns = nonsignificant, * = <0.05, ** = <0.01 , ***<0.001 , ****<0.0001 .

[0022] Figure 5. Prior treatment with carboplatin does not impair cytotoxic immune cell expansion in tumours. A) Dosing regime in UPK10 tumour bearing C57bl / 6 mice. Mice received two doses of carboplatin or PBS prior to injection with a single dose of IL-15SA. Tumours were phenotypes 3 days later. B) Total number of NK cells and CD8 T cells in tumour tissue. C) Total number of splenic NK cells and CD8 T cells. D) Quantification of Ki67+ NK and CD8 T cells in tumour and E) spleen.

[0023] Error bars represent SEM. Statistical tests are one-way anova. n = 20 mice, one independent experiment, p values - ns = nonsignificant, * = <0.05, ** = <0.01 , ***<0.001 , ****<0.0001 .

[0024] Figure 6. Efficacy of combination therapy is dependent on tumour genetics. A) Dosing regimen for ID8p53- / -Brca1- / - tumour bearing albino C57bl / 6 mice. Mice received three cycles of IL-15SA 72 hours priorto carboplatin treatment, single treatments or PBS only. Weekly in vivo imaging was performed to determine tumour burden until clinical endpoint. B) Quantification of in vivo imaging in carboplatin (CP) and carboplatin + IL-15SA combination (Combo) treatment groups on day 53. C) Probability of survival data.

[0025] Error bars represent SEM. n = 40 mice, two independent experiments for each cell line. Error bars represent SEM, Mann Whitney test (B),

[0026] Detailed Description of the Invention

[0027] The embodiments of the invention will now be further described. In the following passages, different embodiments are described. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary.

[0028] Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, pathology, oncology, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. The methods and techniques of the present disclosure are generally performed according to conventional methods well-known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See, e.g., Green and Sambrook et al., Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2012).

[0029] Therapy

[0030] The present inventors have surprisingly found that administering IL-15 significantly enhances the efficacy of chemotherapy for the treatment cancer. In particular, prior treatment with IL-15 or a derivative thereof followed by chemotherapy resulted in significantly reduced tumour growth compared to chemotherapy treatment alone. As such the present invention relates to the combination of IL-15 or a derivative thereof and a chemotherapeutic agent for the treatment of cancer.

[0031] A first aspect of the invention relates to a pharmaceutical composition comprising IL-15 or a derivative thereof, for use in combination with a chemotherapeutic agent in the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent.

[0032] The inventors demonstrate herein that treatment with IL-15 or a derivative thereof prior to treatment with a chemotherapeutic agent enhances the efficacy of the treatment, as evidenced by reduced tumour growth. It is hypothesised that the treatment with IL-15 results in immune cell expansion in the subject wherein the expanded immune cells can enhance the effect of subsequent chemotherapy treatment. Therefore, in an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 minute and 3 months, 1 minute and 2 months, or 1 minute and 1 month prior to said chemotherapeutic agent. In an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 hour and 3 months, 1 hour and 2 months, or 1 hour and 1 month prior to said chemotherapeutic agent. In an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 day and 3 months, 1 day and 2 months, or 1 day and 1 month prior to said chemotherapeutic agent. In an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 minute and 12 weeks, 1 minute and 11 weeks, 1 minute and 10 weeks, 1 minute and 9 weeks, 1 minute and 8 weeks, 1 minute and 7 weeks, 1 minute and 6 weeks, 1 minute and 5 weeks, 1 minute and 4 weeks, 1 minute and 3 weeks, 1 minute and 2 weeks, or 1 minute and 1 week prior to said chemotherapeutic agent. In an embodiment the pharmaceutical composition comprising IL-15 orderivative thereof is administered between 1 hour and 12 weeks, 1 hour and 11 weeks, 1 hour and 10 weeks, 1 hour and 9 weeks, 1 hour and 8 weeks, 1 hour and 7 weeks, 1 hour and 6 weeks, 1 hour and 5 weeks, 1 hour and 4 weeks, 1 hour and 3 weeks, 1 hour and 2 weeks, or 1 hour and 1 week prior to said chemotherapeutic agent. In an embodiment the pharmaceutical composition comprising IL-15 orderivative thereof is administered between 1 day and 12 weeks, 1 day and 11 weeks, 1 day and 10 weeks, 1 day and 9 weeks, 1 day and 8 weeks, 1 day and 7 weeks, 1 day and 6 weeks, 1 day and 5 weeks, 1 day and 4 weeks, 1 day and 3 weeks, 1 day and 2 weeks, or 1 day and 1 week prior to said chemotherapeutic agent. In an embodiment the IL-15 or derivative thereof is administered between 1 minute and 14 days, 1 minute and 13 days, 1 minute and 12 days, 1 minute and 11 days, 1 minute and 10 days, 1 minute and 9 days, 1 minute and 8 days, 1 minute and 7 days, 1 minute and 6 days, 1 minute and 5 days, 1 minute and 4 days, 1 minute and 3 days, 1 minute and 2 days, or 1 minute and 1 day prior to said chemotherapeutic agent. In an embodiment the IL-15 or derivative thereof is administered between 1 hour and 14 days, 1 hour and 13 days, 1 hour and 12 days, 1 hour and 11 days, 1 hour and 10 days, 1 hour and 9 days, 1 hour and 8 days, 1 hour and 7 days, 1 hour and 6 days, 1 hour and 5 days, 1 hour and 4 days, 1 hour and 3 days, 1 hour and 2 days, or 1 hour and 1 day prior to said chemotherapeutic agent. In an embodiment the IL-15 or derivative thereof is administered between 1 day and 14 days, 1 day and 13 days, 1 day and 12 days, 1 day and 11 days, 1 day and 10 days, 1 day and 9 days, 1 day and 8 days, 1 day and 7 days, 1 day and 6 days, 1 day and 5 days, 1 day and 4 days, 1 day and 3 days, or 1 day and 2 days, prior to said chemotherapeutic agent.

[0033] The terms “minute”, “hour”, “day”, “week” and “month” all have their usual meaning in the art.

[0034] IL-15 is an inflammatory cytokine that is encoded by the IL-15 gene. IL-15 plays a key role in the T cell response and the regulation of tissue repair. IL-15 is expressed in association with its high affinity IL-15Ra on the surface of IL15-producing cells and delivers signals to target cells that express IL-2R0 and IL-2Ry receptor subunits. IL-15Ra is the IL-15 receptor alpha subunit which is encoded by the IL15RA gene. In an embodiment the IL-15 is human IL-15 i.e. IL-15 that is derived from human.

[0035] IL-15 may comprise the following sequence (Uniprot P40933)

[0036] SEQ ID NO: 1 MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQS MHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTES GCKECEELEEKNIKEFLQSFVHIVQMFINTS

[0037] In an embodiment the IL-15 or a derivative thereof may comprise SEQ ID NO: 1 or a portion thereof. SEQ ID NO: 1 comprises a signal peptide as such the IL-15 or derivative thereof used in the present invention may or may not comprise said signal peptide. In an embodiment the IL- 15 does not comprise the signal peptide and comprises SEQ ID NO: 2 or SEQ ID NO: 3.

[0038] SEQ ID NO: 2

[0039] NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVEN

[0040] LIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS SEQ ID NO: 3

[0041] MNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVE NLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS

[0042] There are known isoforms of IL-15, as such the IL-15 or derivative thereof may comprise an isoform of IL-15. The IL-15 or derivative thereof may also comprise de novo designed protein mimics of IL-15.

[0043] In some embodiments the IL-15 or derivative thereof comprises a variant of IL-15. A variant of IL-15 refers to IL-15 which comprises a different amino acid at one or more position within the polypeptide chain compared to that of the wild-type IL-15, for example the sequences provided in SEQ ID NO: 1 or SEQ ID NO: 2. The variant IL-15 may comprise a variant amino acid, or amino acid substitution at one or more position of SEQ ID NO:1 , SEQ ID NO: 2 or SEQ ID NO: 3.

[0044] The term “variant amino acid” and “amino acid substitution” are used interchangeably herein and used to refer to any amino acid that is not the amino acid present in the wild-type amino acid sequence, in this case the wild-type IL-15 sequence. The wild-type sequence may be a wildtype human IL-15 sequence. The term “wild-type IL-15 sequence” may refer to the entire sequence or part thereof of SEQ ID NO: 1 , SEQ ID NO: 2, or SEQ ID NO: 3.

[0045] In some embodiments the IL-15 or derivative thereof comprises an amino acid sequence with 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% sequence identity to SEQ ID NO: 1 , SEQ ID NO: 2 or SEQ ID NO: 3.

[0046] The variant amino acid may be a result of substituting, replacing or modifying the original (e.g. wild-type or germline) amino acid, within a protein sequence, with a different amino acid. The process of substituting or replacing an amino acid can be done using standard techniques available to the skilled person, e.g. using recombinant DNA technology. Modification of an amino acid may be performed post-translationally and a variety of chemical or bioconjuagtion methods may be used to modify said amino acid. The amino acids are changed relative to the native (wild type I germline) sequence as found in nature in the wild type (WT). By "wild type" or "WT" or "native" herein is meant an amino acid sequence or a nucleotide sequence that is found in nature, including allelic variations and isoforms. A wild-type protein or polypeptide has an amino acid sequence or a nucleotide sequence that has not been intentionally modified.

[0047] The term "amino acid" as used herein refers to one of the 20 naturally occurring (canonical) amino acids or any non-natural analogues (non-canonical amino acids) that may be present at a specific, defined position within a peptide sequence. “Amino acid” encompasses both naturally occurring and synthetic amino acids. Although in most cases, when the protein is to be produced recombinantly, only naturally occurring amino acids are used. The variant amino acid may comprise one of the twenty canonical amino acids. The variant amino acid may comprise a non- canonical amino acid, also known are non-natural amino acids for example hydroxyproline, hydroxylysine, phosphoserine, phosphothreonine, phosphotyrosine, N-acetyl lysine, methyllysine.

[0048] The terms "polypeptide" and "protein" are used interchangeably to refer to a polymer of amino acid residues and are not limited to a minimum length. Peptides, oligopeptides, dimers, multimers, and the like, are also composed of linearly arranged amino acids linked by peptide bonds, and whether produced biologically, recombinantly, or synthetically and whether composed of naturally occurring or non-naturally occurring amino acids, are included within this definition. Both full-length proteins and fragments thereof are encompassed by the definition. The terms also include co-translational and post-translational modifications of the polypeptide, such as, for example, disulfide-bond formation, glycosylation, acetylation, phosphorylation, proteolytic cleavage (e.g., cleavage by furins or metalloproteases and prohormone convertases (PCs)), and the like. Furthermore, for purposes of the present invention, a "polypeptide" encompasses a protein that includes modifications, such as deletions, additions, substitutions and post-translational modifications (generally conservative in nature as would be known to a person in the art), to the native sequence, as long as the protein maintains the desired activity. These modifications can be deliberate, as through site-directed mutagenesis, or can be accidental, such as through mutations of hosts that produce the proteins, or errors due to PCR amplification or other recombinant DNA methods. Polypeptides or proteins are composed of linearly arranged amino acids linked by peptide bonds, but in contrast to peptides, have a well- defined conformation.

[0049] Proteins, as opposed to peptides, generally consist of chains of 50 or more amino acids. For the purposes of the present invention, the term "peptide" as used herein typically refers to a sequence of amino acids of made up of a single chain of D- or L-amino acids or a mixture of D- and L-amino acids joined by peptide bonds. Generally, peptides contain at least two amino acid residues and are less than about 50 amino acids in length.

[0050] In an embodiment the pharmaceutical composition comprises isolated IL-15 or a derivative thereof. The term "isolated" molecule, protein or polypeptide refers to a molecule, protein or polypeptide that is substantially free of other proteins or polypeptides, having different antigenic specificities. Moreover, protein or polypeptide may be substantially free of other cellular material and / or chemicals. Thus, the protein, nucleic acids and polypeptides described herein are preferably isolated. Thus, as used herein, an "isolated" protein, or polypeptide means protein or polypeptide that has been identified and separated and / or recovered from a component of its natural cell culture environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the protein or polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.

[0051] As discussed above the IL-15 or derivative thereof may comprise one or more amino acid substitution or variant amino acid provides beneficial properties to the protein for example enhanced activity or binding. In some embodiments the IL-15 or derivative thereof comprises an amino acid substitution that confers super agonist activity to the IL-15. In an embodiment the IL- 15 or derivative thereof comprises an IL-15 super agonist. In an embodiment the IL-15 or derivative thereof comprises an IL-15 super agonist mutation i.e., an amino acid substitution or variant amino acid that confers super agonist activity. The skilled person will be able to determine suitable amino acid substitutions which confer super agonist activity using standard mutational and screening techniques known in the art.

[0052] Suitable IL-15 super agonist mutations are known in the art for example the super agonist mutation may comprise N72D (IL-15N72D), wherein the amino acid positions are numbered according to SEQ ID NO.2. Further super agonist mutations may be identified by the skilled person using methods known in the art for example structure guided mutagenesis.

[0053] IL-15 is expressed in association with its high affinity IL-15Ra as such in an embodiment the IL- 15 or derivative thereof comprises a complex of IL-15 and IL-15Ra. In an embodiment the pharmaceutical composition comprises a complex of IL-15 and IL-15Ra. The IL-15 may be provided in complex with the full-length IL-15Ra or a portion thereof, for example the IL-15Ra sushi domain (IL-15RaSu). The IL-15Ra sushi domain is the extracellular portion of the IL-15Ra. The portion of the IL-15Ra may comprise the IL-15Ra sushi domain and may further comprise additional domains from the full-length IL-15 receptor for example the linker domain, the proline / threonine rich domain, the transmembrane domain, the cytoplasmic tail or the hinge region from the full-length receptor. The IL-15 or derivative thereof may be provided as a complex with IL-15Ra before being administered to a subject in need thereof. The IL-15 or derivative thereof and the IL-15-Ra may be provided separately and then combined and allowed to form a complex before being administered to a subject in need thereof. The IL-15 or derivative thereof and the IL-15-Ra may be provided as a fusion protein. The IL-15 or derivative thereof may be linked to the IL-15-Ra via a peptide linker. The peptide linker may be a flexible linker, a rigid linker or a cleavable linker. Suitable flexible peptide linkers are known in the art such as Gly / Ser linkers. Suitable rigid linkers are known in the art such as alpha helix-forming linkers or proline rich linkers. Suitable cleavable linkers are known in the art such as protease cleavable linkers. The IL-15 or derivative thereof or the IL-15Ra may comprise a post-translational modification.

[0054] Post-translational modifications may be selected from phosphorylation and / or glycosylation.

[0055] In order to extend the half-life of the IL-15 within a subject the IL-15 may comprise a half-life extending moiety. The pharmaceutical composition may comprise a half-life extending moiety. The IL-15 or a derivative thereof may comprise or a half-life extending moiety, for example the half-life extending moiety may be conjugated to the IL-15 or a derivative thereof. Various moieties and approaches are known for extending the half-life of protein-based therapeutics and any half-life extending approach may be used in the present invention. In an embodiment the half-life extending moiety may be selected from the following non-limiting list: a human immunoglobulin Fc domain, polyethylene glycol (PEG), PEG derivatives, simple lipids, lipid dicarboxylic acids, lipids with additional moieties, human serum albumin binders, e.g. smallmolecule binders or antibodies / antibody fragments that bind human serum albumin, human serum albumin, or streptococcal protein G’s albumin-binding domain (ABD). Examples of lipids include glucagon-like peptide 1 (GLP-1), the analogs GLP-1 liraglutide and semaglutide or cholesterol. In a preferred embodiment the half-life extending moiety is selected from one or more of an Fc domain, a modified Fc domain, PEG, serum albumin, a serum albumin binding molecule. The half-life extending moiety may be conjugated to the IL-15 or a derivative thereof. The half-life extending moiety may be conjugated to the IL-15Ra or portion thereof. The half-life extending moiety may be conjugated via a peptide linker.

[0056] In some embodiments the half-life extending moiety is an Fc domain. In an embodiment the Fc domain is a human IgG Fc domain. In an embodiment the Fc domain is a human lgG1 Fc domain.

[0057] By "Fc" or "Fc region" or "Fc domain" as used herein is meant the polypeptide comprising the constant region of an antibody excluding the first constant region immunoglobulin domain (CH1) and, in some cases, part of the hinge. In one embodiment, the Fc domain includes constant region immunoglobulin domains CH2, CH3 and the hinge region between CH1 and CH2 or part of the hinge region.

[0058] In human, Fc refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, the last three constant region immunoglobulin domains of IgE and IgM, and the flexible hinge N-terminal to these domains. For IgA and IgM, Fc may include the J chain. For IgG, the Fc domain comprises immunoglobulin domains CH2 and CH3 and the lower hinge region between CH1 and CH2. Although the boundaries of the Fc region may vary, the human IgG heavy chain Fc region is usually defined to include residues C226 or P230 to its carboxylterminus, wherein the numbering is according to the EU index as in Kabat. Fc as used herein may refer to the Fc region in isolation, or this region in the context of an Fc fusion ("fusion composition" or "fusion construct"). Fc domains include all or part of an Fc region; that is, N- or C- terminal sequences may be removed from wild-type or variant Fc domains, as long as this does not affect function.

[0059] The Fc may be modified in order to further improve half-life extension. A number of approaches to modify Fc half-life are known in the art, for example the Fc region may be modified to enhance interaction with FcRn receptor and thereby improve half-life of the 11-15 or derivative thereof. IgG naturally persists for a prolonged period in the serum due to FcRn-mediated recycling, giving it a typical half-life of approximately 21 days. Half-life can be extended by engineering the pH- dependant interaction of the Fc domain with FcRn to increase affinity at pH 6.0 while retaining minimal binding at pH 7.4. Examples of such engineered Fc include: the T250Q / M428L variant, which conferred an approximately 2-fold increase in IgG half-life (assessed in rhesus monkeys); the M252Y / S254T / T256E variant, which gave an approximately 4-fold increase in IgG half-life (assessed in cynomolgus monkeys). The M252Y / S254T / T256E mutation commonly known as “YTE” variant, comprises mutations in lgG1 CH2 and is an approach used in the art to extend half-life of an Fc domain. Extending half-life may allow the possibility of decreasing administration frequency, while maintaining or improving efficacy. The Fc region may be modified to reduce interaction with FcyR receptor and thereby improve half-life of the IL-15 or derivative thereof.

[0060] The half-life extending moiety may be conjugated to the IL-15, the IL-15 Ra or both. As such the IL-15 or a derivative thereof may comprise a fusion protein of one or more of: a fusion protein of IL-15 or a derivative thereof and a half-life extending moiety e.g. an Fc domain; a fusion protein of IL-15Ra or a portion thereof and a half-life extending moiety e.g. an Fc domain. In an embodiment the half-life extending moiety is an lgG1 Fc and is conjugated to the IL-15Ra or a portion thereof. In a preferred embodiment the IL-15Ra is provided as an IL-15RaSu / lgG1 Fc fusion protein.

[0061] The pharmaceutical composition used in the present invention advantageously may comprise an IL-15 super agonist as described herein in complex with IL-15Ra or a portion thereof and a half-life extension moiety. In some embodiments the composition may comprise one or more copies of the IL-15 or a derivative thereof, the IL-15Ra or a portion thereof, and the half-life extension moiety. In a particular embodiment the IL-15 or derivative thereof comprises a dimeric IL-15RaSu / lgG1 Fc and two IL-15N72D molecules.

[0062] The IL-15 or derivative thereof may be selected from one or more of ALT803 also referred to as N803, XmAb306, NKTR-225, RLI (Receptor Linker) IL-15 also referred to as SO-C101 , rhlL-15, hetlL-15 also referred to as NIZ985, NL-201 , IL-15RaA2, IL-15RaA3, IL-15RalC3, IL-15A6, IL- 15LSP, sch rhlL-15. ALT803 comprises an IL-15 variant with a higher binding affinity (IL- 15N72D) complexed with a human IL-15Ra sushi domain-Fc fusion protein. XmAb306 is a IL15 / IL15F?a complex comprising an engineered Fc domain. NKTR-225 comprises a polyethylene glycol-conjugate of rhlL-15 designed to have a high binding affinity to IL-15R and increased persistence. RLI I SO-C101 comprises an IL-15 agonist fusion protein, comprising the IL-15Ra sushi domain fused to IL-15 through a 20 amino acid linker. hetlL-15 (NIZ985) comprises a fully glycosylated form of hetlL-15 where the C-terminus of IL-15Ra is covalently linked to the Fc region of human lgG1. rhlL-15 comprises recombinant human IL-15. NL-201 comprises a de novo designed mimic of IL-15. IL-15RaA2 comprises the deletion of the sushi domain IL-15Ra. IL-15RaA3 comprises deletion of the linker / hinge region of IL-15Ra. Sch rhlL- 15 comprises an Escherichia coli-demed IL-15 monomer.

[0063] The present invention relates to the combination of IL-15 or a derivative thereof and a chemotherapeutic agent for the treatment of cancer, as such the chemotherapeutic agent may be any suitable chemotherapeutic. In an embodiment the chemotherapeutic agent is selected from a platinum-based chemotherapeutic agent, an anthracycline-based chemotherapeutic agent, and alkylating chemotherapeutic agent, a taxane-based chemotherapeutic agent, a vinca alkaloid-based chemotherapeutic agent, a retinoid-based chemotherapeutic agent, a histone deacetylase inhibitor, a topoisomerase I inhibitor, a topoisomerase II inhibitor, a kinase inhibitor, or a nucleotide analogue-based chemotherapeutic agent. Examples of the aforementioned chemotherapeutic agents are known to the skilled person and the skilled person would be able to select a suitable agent.

[0064] In an embodiment the chemotherapeutic is a platinum-based chemotherapeutic agent. The platinum-based chemotherapeutic agent may be selected from carboplatin, cisplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin and / or combinations thereof. Ina preferred embodiment the platinum-based chemotherapeutic agent is carboplatin.

[0065] The present invention provides a treatment for cancer. In an embodiment the cancer may be selected from ovarian cancer, pancreatic cancer, bone cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, breast cancer, brain cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the oesophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, kidney cancer, sarcoma of soft tissue, cancer of the urethra, cancer of the bladder, renal cancer, lung cancer, non-small cell lung cancer, thymoma, urothelial carcinoma leukaemia, prostate cancer, omentum cancer, mesothelioma, adrenocortical carcinoma, lymphomas, such as such as Hodgkin's disease, non-Hodgkin's, gastric cancer, and multiple myelomas. In a specific embodiment the cancer is ovarian cancer.

[0066] The amount of the pharmaceutical composition comprising IL-15 or a derivative thereof and the chemotherapeutic agent that is effective / active in the treatment of a particular cancer will depend on the nature of the disease or condition and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays can optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the compositions will also depend on the route of administration, and the seriousness of the disease or disease, and should be decided according to the judgment of the practitioner and each patient's circumstances. Factors like age, body weight, sex, diet, time of administration, rate of excretion, condition of the host, drug combinations, reaction sensitivities and severity of the disease shall be taken into account.

[0067] The pharmaceutical composition comprising IL-15 or derivative thereof may be administered at any suitable dose. As an example the pharmaceutical composition comprising IL-15 or derivative thereof may be administered is administered at a dose of between 0.1 pg / kg and 100 mg / kg,

[0068] 1 pg / kg and 100 mg / kg, 10pg / kg and 100 mg / kg, 50pg / kg and 100 mg / kg, 100pg / kg and 100 mg / kg, 500pg / kg and 100 mg / kg, 750pg / kg and 100 mg / kg, 1 mg / kg and 100 mg / kg, 10mg / kg and 100 mg / kg, 50mg / kg and 100 mg / kg, 75mg / kg and 100 mg / kg, 0.1 pg / kg and 50 mg / kg, 0.1 pg / kg and 10 mg / kg, 0.1 pg / kg and 1 mg / kg.

[0069] The present invention relates to the combination of IL-15 or a derivative thereof and a chemotherapeutic agent, the present inventors have shown that it is advantageous to administer the IL-15 or derivative thereof prior to treatment with a chemotherapeutic. As such the prior treatment with a pharmaceutical composition comprising IL-15 may comprise a single dose or multiple doses. In an embodiment wherein multiple doses of the pharmaceutical composition comprising IL-15 or derivative thereof are administered prior to said chemotherapeutic agent. Where multiple doses of the pharmaceutical composition comprising IL-15 or derivative thereof are provided the doses are provided at separate intervals, where there is a time period in between the administrations of the doses. The time period between each dose of the pharmaceutical composition comprising IL-15 or derivative thereof may be between 1 minute and 7 days, 1 minute and 6 days, 1 minute and 5 days, 1 minute and 4 days, 1 minute and 3 days, 1 minute and 2 days, 1 minute and 1 day, 1 hour and 7 days, 1 hour and 6 days, 1 hour and 5 days, 1 hour and 4 days, 1 hour and 3 days, 1 hour and 2 days, 1 hour and 1 day, 1 day and 7 days, 1 day and 6 days, 1 day and 5 days, 1 day and 4 days, 1 day and 3 days, 1 day and

[0070] 2 days. The time period between each dose may be the same or different.

[0071] A treatment cycle as used in the present invention may comprise a single dose of the pharmaceutical composition comprising IL-15 or derivative thereof followed by a therapy with a suitable chemotherapeutic agent as described herein. The dose and dosage schedule of the chemotherapeutic may be the approved dose. The treatment cycle may comprise multiple doses of the pharmaceutical composition comprising IL-15 or derivative thereof followed by a therapy with a suitable chemotherapeutic agent as described herein. The treatment cycles may be repeated and adjusted accordingly.

[0072] In embodiments the pharmaceutical composition comprising IL-15 or derivative thereof may be administered via any suitable route for example systemically, intravenously, subcutaneously, intramuscularly, intravesically, or orally.

[0073] In embodiments the chemotherapeutic agent may be administered via any suitable route for example systemically, intravenously, subcutaneously, intramuscularly, intravesically, intrathecally or orally. Depending on the chemotherapeutic agent that is used the approved administration route may be used.

[0074] The pharmaceutical composition comprising IL-15 or derivative thereof may be administered as a bolus dose or as a continuous infusion. The term “bolus dose” refers to a discrete dosage which is administered to a subject in a short time frame for example 1 minute to 1 hour. The term “continuous infusion” refers to using intravenous administration over a prolonged period for example more than 1 day. Where a single dose of the pharmaceutical composition comprising IL-15 is provided prior to treatment with the chemotherapeutic agent, the single dose may be provided as a bolus dose or continuous infusion. Where multiple doses of the pharmaceutical composition comprising IL-15 is provided prior to treatment with the chemotherapeutic agent, the multiple doses may be provided as bolus doses or continuous infusion, or a mixture thereof.

[0075] The pharmaceutical composition comprising IL-15 or a derivative thereof, may be provided with any pharmaceutically acceptable excipient, diluent or carrier. The pharmaceutically acceptable carrier or vehicle can be particulate, so that the compositions are, for example, in tablet or powder form. The term "carrier" refers to a diluent, adjuvant or excipient, with which a pharmaceutical composition of the present invention is administered. Such pharmaceutical carriers can be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. The carriers can be saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliary, stabilizing, thickening, lubricating and coloring agents can be used. In one embodiment, when administered to a subject, the polypeptide of the present invention or compositions and pharmaceutically acceptable carriers are sterile. Water is a preferred carrier when the pharmaceutical composition of the present invention are administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The present compositions, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents.

[0076] The pharmaceutical composition can be in the form of a liquid, e.g., a solution, syrup, solution, emulsion or suspension. The liquid can be useful for oral administration or for delivery by injection, infusion (e.g., IV infusion) or sub-cutaneous.

[0077] When intended for oral administration, the composition can be in solid or liquid form, where semisolid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.

[0078] As a solid composition for oral administration, the composition can be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form. Such a solid composition typically contains one or more inert diluents. In addition, one or more of the following can be present: binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, corn starch and the like; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavoring; and a coloring agent. When the composition is in the form of a capsule (e. g. a gelatin capsule), it can contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol, cyclodextrin or a fatty oil.

[0079] When intended for oral administration, a composition can comprise one or more of a sweetening agent, preservatives, dye / colorant and flavor enhancer. In a composition for administration by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent can also be included.

[0080] Compositions can take the form of one or more dosage units.

[0081] In specific embodiments, it can be desirable to administer the composition locally to the area in need of treatment, or by intravenous injection or infusion.

[0082] The amount of the polypeptide, fusion protein or pharmaceutical composition described herein that is effective / active in the treatment of a particular disease or condition will depend on the nature of the disease or condition and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays can optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the compositions will also depend on the route of administration, and the seriousness of the disease or disease, and should be decided according to the judgment of the practitioner and each patient's circumstances. Factors like age, body weight, sex, diet, time of administration, rate of excretion, condition of the host, drug combinations, reaction sensitivities and severity of the disease shall be taken into account.

[0083] Typically, the amount is at least about 0.01 % of a polypeptide of the present invention by weight of the composition. When intended for oral administration, this amount can be varied to range from about 0.1 % to about 80% by weight of the composition. Preferred oral compositions can comprise from about 4% to about 50% of the polypeptide of the present invention by weight of the composition.

[0084] Compositions can be prepared so that a parenteral dosage unit contains from about 0.01 % to about 2% by weight of the polypeptide of the present invention.

[0085] For administration by injection, the composition can comprise from about typically about 0.1 mg / kg to about 250 mg / kg of the subject’s body weight, preferably, between about 0.1 mg / kg and about 20 mg / kg of the subject’s body weight, and more preferably about 1 mg / kg to about 10 mg / kg of the subject’s body weight. In one embodiment, the composition is administered at a dose of about 1 to 30 mg / kg, e.g., about 5 to 25 mg / kg, about 10 to 20 mg / kg, about 1 to 5 mg / kg, or about 3 mg / kg. The dosing schedule can vary from e.g., once a week to once every 2, 3, or 4 weeks or more.

[0086] The pharmaceutical composition comprising IL-15 or a derivative thereof may be used in combination with any further anti-cancer agent or standard of care treatment. In an embodiment the pharmaceutical composition comprising IL-15 or a derivative thereof is used in combination with a chemotherapeutic agent, and a further anti-cancer agent or standard of care treatment may also be administered. In an embodiment the further anti-cancer agent is selected from immunotherapy, hormone therapy, biologic, cytokine, small molecule, gene therapy, viral therapy, bone marrow transplantation, nanotherapy, targeted anti-cancer therapies, oncolytic drugs, therapeutics which target DNA damage repair and homologous recombination deficiency, CAR-T therapy and / or autologous stem cell rescue (ASCR). Examples of therapeutic agents which may be used as a further anti-cancer agent include other checkpoint inhibitors, antineoplastic agents, immunogenic agents, attenuated cancerous cells, tumor antigens, antigen presenting cells such as dendritic cells pulsed with tumor-derived antigen or nucleic acids, immune stimulating cytokines (e.g., IL-2, IFNa2, GM-CSF), targeted small molecules and biological molecules (such as components of signal transduction pathways, e.g. modulators of tyrosine kinases and inhibitors of receptor tyrosine kinases, and agents that bind to tumorspecific antigens, including EGFR antagonists), an anti-inflammatory agent, a cytotoxic agent, a radiotoxic agent, or an immunosuppressive agent and cells transfected with a gene encoding an immune stimulating cytokine (e.g., GM-CSF), chemotherapy. A biologic may be an antibody therapy, for example an antibody that targets a checkpoint inhibitor, such as PD-1 (e.g. Pembrolizumab, Nivolumab or Cemiplimab), PD-L1 (e.g. Atezolizumab, Avelumab or Durvalumab), PD-L2, LAG-3 (e.g. Relatlimab), Tim-3 or CTLA4 (e.g. Ipilimumab). The further anti-cancer therapy may comprise surgery.

[0087] The anti-cancer treatment of the present invention may be provided to a subject who is treatment naive i.e. has not received any prior treatment. The anti-cancer treatment of the present invention may be provided to a subject who has received prior treatment for said cancer. Prior treatment for said cancer may involve any anti-cancer therapy for example surgery, immunotherapy, hormone therapy, biologic, cytokine, small molecule, gene therapy, viral therapy, bone marrow transplantation, nanotherapy, targeted anti-cancer therapies, oncolytic drugs, CAR-T therapy, therapeutics which target DNA damage repair and homologous recombination deficiency and / or autologous stem cell rescue (ASCR). In an embodiment the subject received one or more further anti-cancer agent or anti-cancer treatment prior to the administration of the pharmaceutical composition comprising IL-15 or a derivative thereof in accordance with the present invention. The subject may have shown a response to the prior anti-cancer therapy. The subject may not have shown any measurable response to the prior anti-cancer therapy. In an embodiment the cancer is relapsed or refractory.

[0088] Without wishing to be bound by theory, it is hypothesised that the present invention allows the expansion of immune cells which can then enhance the effect of the chemotherapeutic agent, as such in an embodiment the pharmaceutical composition comprising IL-15 or a derivative thereof increases the level of natural killer (NK) cells and / or CD8+ T cells, present in a subject.

[0089] In an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered prior to the chemotherapeutic agent, wherein after administration of the pharmaceutical composition but prior to administration of the chemotherapeutic, the subject is assessed forthe level of NK cells and / or CD8+ T cells. In an embodiment the subject is assessed for an increased level of NK cells and / or CD8+ T cells, indicating they are suitable for therapy with the chemotherapeutic. In an embodiment the pharmaceutical composition comprising IL-15 or derivative thereof is administered prior to the chemotherapeutic agent, wherein after administration of the pharmaceutical composition but prior to administration of the chemotherapeutic, the subject is assessed for the level of immune cell activation. Based on the level of immune cell activation the subject may be selected for further treatment with said chemotherapeutic. Immune cell activation may be assessed using various parameters, for example level of Ki67 expression, interferon gamma, granzyme B and / or perforin expression. In an embodiment the subject is assessed for the level of immune cell activation for example by identifying higher level of expression of Ki67. In an embodiment the subject is assessed for an increased level of immune cell activation for example by identifying higher level of Ki67 expression, interferon gamma, granzyme b and / or perforin expression. The increased level of NK cells, CD8+ T cells, Ki67 expression, interferon gamma, granzyme b and / or perforin expression may be assessed by comparing to a reference value. In certain embodiments the level of NK cells, CD8+ T cells, expression of Ki67 is increased, decreased or the same as the reference value. The level of immune cell activation may be assessed ex vivo and / or after restimulation. The subject may be assessed for the level of NK cells, CD8+ T cells, and / or immune activation via an in vitro or ex vivo assay. In some embodiments a sample is obtained from said subject and the level of NK cells, CD8+ T cells, and / or immune activation is determined.

[0090] In an embodiment, prior to treatment with IL-15 and a chemotherapeutic agent the subject is assessed for the level of NK cells and / or CD8+ T cells. In an embodiment the level of NK cells and / or CD8+ T cells present in the subject is compared to a reference value. The level of NK cells and / or CD8+ T cells present in the subject may be increased, decreased or the same as the reference value and may indicate that the subject is suitable for therapy IL-15 and a chemotherapeutic. In an embodiment prior to treatment with IL-15 and a chemotherapeutic agent the level of immune cell activation is assessed, for example by identifying the level of expression of Ki67. In an embodiment the level of expression of Ki67 in the subject is compared to a reference value. The level of expression of Ki67present in the subject may be increased, decreased or the same as the reference value and may indicate that the subject is suitable for therapy IL-15 and a chemotherapeutic.

[0091] The reference value may be obtained from a healthy individual, a diseased individual, or the subject undergoing therapy.

[0092] In order to assess the level of NK cell and / or CD8+ T cells a biological sample may be obtained or provided by the subject. The biological sample may be tissue, tumour, saliva, urine, blood including whole blood and plasma. Where a tissue sample is obtained the tissue may be selected from spleen, omentum, pancreas, lung. The biological sample may comprise tumour tissue, cancer cells and / or healthy tissue.

[0093] In some embodiments the treatment comprises a step of measuring the level of NK cells, CD8+ T cells, expression of Ki67. The step of measuring may be performed using any suitable method.

[0094] As will be appreciated by the skilled person, the terms “treating”, “treats” and “treatment” include both preventative and curative treatment of a condition, disease or disorder. These terms also include slowing, interrupting, controlling or stopping the progression of a condition, disease or disorder and preventing, curing, slowing, interrupting, controlling or stopping the symptoms of a condition, disease or disorder. As used herein, "treat", "treating" or "treatment" means inhibiting or relieving a disease or disease. For example, treatment can include a postponement of development of the symptoms associated with a disease or disease, and / or a reduction in the severity of such symptoms that will, or are expected, to develop with said disease. The terms include ameliorating existing symptoms, preventing additional symptoms, and ameliorating or preventing the underlying causes of such symptoms. Thus, the terms denote that a beneficial result is being conferred on at least some of the mammals, e.g., human or canine patients, being treated. Many medical treatments are effective for some, but not all, patients that undergo the treatment.

[0095] The term "subject" or "patient" refers to an animal or human which is the object of treatment, observation, or experiment. By way of example only, a subject includes, but is not limited to, a mammal, including, but not limited to, a human or a non-human mammal, such as a non-human primate, murine, bovine, equine, canine, ovine, or feline.

[0096] The present treatment may result in a complete response or a partial response to therapy. In one embodiment, post-treatment, the subject has at least 7 days, or at least 14 days, or at least 21 days, or at least 28 days, or at least 40 days, or at least 50 days, or at least 60 days disease progression-free. In one embodiment, post-treatment, the subject has at least 7 days, or at least 14 days, or at least 21 days, or at least 28 days, or at least 40 days, or at least 50 days, or at least 60 days disease progression-free.

[0097] In one embodiment, the number of days of survival, the number of disease free days, or the number of disease-progression free days is at least 2 months, or at least 3 months, or at least 4 months, e.g. at least 5 months, such as at least 6 months.

[0098] In one embodiment, the number of days of survival, the number of disease free days, or the number of disease-progression free days is at least 9 months, 200 days, 300 days or 3 years or more. In one embodiment, it is least one, two, three or more years.

[0099] In one embodiment, the subject has increased expression, amplification of expression and / or mutation in the KRAS gene.

[0100] In one embodiment, the subject has reduced expression, loss of expression and / or a mutation in the p53 gene.

[0101] In one embodiment, the subject has reduced expression, loss of expression and / or a mutation in a Brea gene. The Brea gene may selected from the Brcal or Brca2 gene. The term “increased expression” or “increased level” may refer to an increase compared to a reference sample. The reference sample may be any suitable reference sample for example from a wild-type sample or healthy subject. The reference value may be obtained from the present subject prior to treatment or prior to development of disease. The increase may comprise 1 %, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% increase relative to the reference sample. The term “decreased expression” or “decreased level” may refer to a decrease compared to a reference sample. The reference sample may be any suitable reference sample for example from a wild-type sample or healthy subject. The reference value may be obtained from the present subject prior to treatment or prior to development of disease The decrease may comprise 1 %, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% decrease relative to the reference sample.

[0102] An aspect of the present invention relates to a method of treating cancer in a subject, comprising administering a pharmaceutical composition comprising IL-15 or a derivative thereof, in combination with a chemotherapeutic agent to said subject, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent.

[0103] The method of treating cancer may comprise any of the further features described herein.

[0104] An aspect of the present invention relates to the use of a pharmaceutical composition comprising IL-15 or a derivative thereof, for the manufacture of a medicament for the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to a chemotherapeutic agent.

[0105] The use of a pharmaceutical composition comprising IL-15 or a derivative thereof, for the manufacture of a medicament for the treatment of cancer may comprise any of the further features described herein.

[0106] Further Methods

[0107] An aspect of the invention relates to an in vitro, ex vivo or in vivo method of sensitising cells to a chemotherapeutic agent comprising: contacting a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, contacting said population of cells to a chemotherapeutic agent. An aspect of the invention relates to an in vitro, ex vivo or in vivo method of enhancing efficacy of a chemotherapeutic agent comprising: contacting a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, contacting said population of cells to a chemotherapeutic agent.

[0108] In some embodiments the population of cells are contacted with a pharmaceutical composition comprising IL-15 or a derivative thereof prior to contacting with the chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition prior to the chemotherapeutic agent. In an embodiment the population of cells is administered between 1 minute and 3 months, 1 minute and 2 months, or 1 minute and 1 month prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 hour and 3 months, 1 hour and 2 months, or 1 hour and 1 month prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 day and 3 months, 1 day and 2 months, or 1 day and 1 month prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 minute and 12 weeks, 1 minute and 11 weeks, 1 minute and 10 weeks, 1 minute and 9 weeks, 1 minute and 8 weeks, 1 minute and 7 weeks, 1 minute and 6 weeks, 1 minute and 5 weeks, 1 minute and 4 weeks, 1 minute and 3 weeks, 1 minute and 2 weeks, or 1 minute and 1 week prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 hour and 12 weeks, 1 hour and 11 weeks, 1 hour and 10 weeks, 1 hour and 9 weeks, 1 hour and 8 weeks, 1 hour and 7 weeks, 1 hour and 6 weeks, 1 hour and 5 weeks, 1 hour and 4 weeks, 1 hour and 3 weeks, 1 hour and 2 weeks, or 1 hour and 1 week prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 day and 12 weeks, 1 day and 11 weeks, 1 day and 10 weeks, 1 day and 9 weeks, 1 day and 8 weeks, 1 day and 7 weeks, 1 day and 6 weeks, 1 day and 5 weeks, 1 day and 4 weeks, 1 day and 3 weeks, 1 day and 2 weeks, or 1 day and 1 week prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 minute and 14 days, 1 minute and 13 days, 1 minute and 12 days, 1 minute and 11 days, 1 minute and 10 days, 1 minute and 9 days, 1 minute and 8 days, 1 minute and 7 days, 1 minute and 6 days, 1 minute and 5 days, 1 minute and 4 days, 1 minute and 3 days, 1 minute and 2 days, or 1 minute and 1 day prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 hour and 14 days, 1 hour and 13 days, 1 hour and 12 days, 1 hour and 11 days, 1 hour and 10 days, 1 hour and 9 days, 1 hour and 8 days, 1 hour and 7 days, 1 hour and 6 days, 1 hour and 5 days, 1 hour and 4 days, 1 hour and 3 days, 1 hour and 2 days, or 1 hour and 1 day prior to said chemotherapeutic agent. In an embodiment the population of cells is contacted with the pharmaceutical composition between 1 day and 14 days, 1 day and 13 days, 1 day and 12 days, 1 day and 11 days, 1 day and 10 days, 1 day and 9 days, 1 day and 8 days, 1 day and 7 days, 1 day and 6 days, 1 day and 5 days, 1 day and 4 days, 1 day and 3 days, or 1 day and 2 days, prior to said chemotherapeutic agent.

[0109] As will be apparent to the skilled person the methods described herein may be performed in vivo, in vitro or ex vivo as appropriate.

[0110] An aspect of the invention relates to a method of identifying a subject as suitable for therapy with IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the IL-15 is administered prior to said chemotherapeutic agent, wherein the method comprises: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for therapy.

[0111] In an embodiment the subject is assessed for an increased level of NK cells and / or CD8+ T cells compared to a reference value, indicating the subject is suitable for therapy. In an embodiment the subject is assessed for NK cells with an increased level of immune cell activation for example by identifying higher level of expression of Ki67 on said NK cells. The level of immune activation may be assessed compared to a reference value.

[0112] In an embodiment the method of identifying a subject a suitable for therapy may be performed before any treatment has started. In an embodiment the method of identifying a subject a suitable for therapy may be performed during or part-way through treatment, wherein the method is for identifying a subject as suitable for further treatment. For example the method may comprise: administering a subject with cancer IL-15 or a derivative thereof, screening a biological sample obtained from said subject with cancer for the presence of NK cells and / or CD8+ T cells, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for further therapy with a chemotherapeutic agent.

[0113] An embodiment relates to method of identifying a subject as suitable for further therapy with a chemotherapeutic agent wherein the subject has received therapy with IL-15 or a derivative thereof comprising: screening a biological sample obtained from said subject with cancer for the presence of NK cells and / or CD8+ T cells, wherein said subject has received therapy with IL-15 or a derivative thereof, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for further therapy with a chemotherapeutic agent.

[0114] The biological sample obtained or provided by the subject with cancer may be tissue, saliva, urine, blood including whole blood and plasma. Where a tissue sample is obtained the tissue may be selected from spleen, omentum, pancreas, lung.

[0115] The biological sample may comprise tumour tissue, tissue comprising cancer cells and / or healthy tissue obtained for the subject.

[0116] The method may further comprise screening the biological sample for the level of CD8+ T cells.

[0117] The threshold level of NK cells and / or CD8+ T cells may be a reference value. The threshold level or reference value may be the level of NK cells and / or CD8+ T cells present in a healthy individual, or a diseased individual. The reference value may be obtained from the present subject prior to treatment. In particular where the level of NK cells and / or CD8+ T cells is assessed after treatment with IL-15 but before treatment with a chemotherapeutic agent the reference value for the NK cells and / or CD8+ T cells may be obtained from the present subject prior to treatment.

[0118] The level of NK cells and / or CD8+ T cells may be assessed in one more of tumour tissue, tissue comprising cancer cells and / or healthy tissue obtained for the subject.

[0119] An aspect of the invention relates to a method of therapy monitoring comprising: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, comparing the level of NK cells and / or CD8+ T cells with a reference value, determining whether the therapy is effective based on the level of NK cells and / or CD8+ T cells compared to said reference value, wherein the subject has received, is currently receiving or will receive a therapy comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the IL-15 is administered prior to said chemotherapeutic agent. The level of NK cells may indicate that the therapy is effective in the subject. Effective therapy may comprise a response to therapy for example a complete response or a partial response. Effective therapy may comprise progression-free disease. In one embodiment, effective therapy may comprise post-treatment, the subject has at least 7 days, or at least 14 days, or at least 21 days, or at least 28 days, or at least 40 days, or at least 50 days, or at least 60 days disease progression-free. In one embodiment, post-treatment, the subject has at least 7 days, or at least 14 days, or at least 21 days, or at least 28 days, or at least 40 days, or at least 50 days, or at least 60 days disease progression-free.

[0120] The presence of NK cells may refer to the presence of NK1 , NK2, NK3 and / or NK4 cells. The level of NK cells that is assessed may comprise the level of NK1 , NK2, NK3 and / or NK4 cells. In an embodiment the NK cells comprise NK1 , NK2, NK3 and / or NK4 cells. In an embodiment the level of NK3 and / or NK4 cells is assessed and compared to a reference value or a threshold value.

[0121] The biological sample obtained or provided by the subject with cancer may be tissue, saliva, urine, blood including whole blood and plasma. Where a tissue sample is obtained the tissue may be selected from spleen, omentum, pancreas, lung. The biological sample may comprise tumour tissue, tissue comprising cancer cells and / or healthy tissue obtained for the subject.

[0122] As shown herein the inventors have demonstrated that the present combination therapy is effective in ID8p53- / -Brca1 - / - tumours, as such the therapy may be particularly effective in subjects with certain genetic characteristics, for example subjects with increased or mutated KRAS, reduced or mutated p53, or reduced or mutated Brea. Therefore, an aspect of the invention relates to a method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises identifying the subject as having one of more of the following; increased expression, amplification of expression and / or mutation in the KRAS gene, reduced expression, loss of expression and / or a mutation in the p53 gene, reduced expression, loss of expression and / or a mutation in a Brea gene, and selecting said subject for therapy.

[0123] Identifying a subjected as having one or more of the above-mentioned genetic characteristics may be performed by obtaining a biological sample from said subject and screening the biological sample for one or more of; increased expression, amplification of expression and / or mutation in the KRAS gene, reduced expression, loss of expression and / or a mutation in the p53 gene, reduced expression, loss of expression and / or a mutation in a Brea gene,

[0124] The genetic information may also be obtained from a previously performed genetic screening test or from a database.

[0125] Kits

[0126] An aspect of the invention relates to a kit for the treatment of cancer comprising IL-15 or a derivative thereof and a chemotherapeutic agent.

[0127] The kit may comprise a pharmaceutical composition comprising IL-15 or a derivative thereof and the chemotherapeutic agent formulated separately as distinct compositions. The kit may comprise the IL-15 or a derivative thereof formulated in a pharmaceutical composition as described herein. The kit may comprise chemotherapeutic agent formulated in a pharmaceutical composition as described herein. The pharmaceutical composition comprising IL-15 or a derivative thereof and the chemotherapeutic agent may be formulated for a specific administration route. The kit may further comprise instructions for use. The kit may further comprise a further anti-cancer treatment as described herein.

[0128] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. While the foregoing disclosure provides a general description of the subject matter encompassed within the scope of the present disclosure, including methods, as well as the best mode thereof, of making and using this disclosure, the following examples are provided to further enable those skilled in the art to practice this disclosure. However, those skilled in the art will appreciate that the specifics of these examples should not be read as limiting on the invention, the scope of which should be apprehended from the claims and equivalents thereof appended to this disclosure. Various further aspects and embodiments of the present disclosure will be apparent to those skilled in the art in view of the present disclosure.

[0129] All documents mentioned in this specification are incorporated herein by reference in their entirety, including references to gene accession numbers, scientific publications and references to patent publications.

[0130] "and / or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. For example "A and / or B" is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein. Unless context dictates otherwise, the descriptions and definitions of the features set out above are not limited to any particular aspect or embodiment of the invention and apply equally to all aspects and embodiments which are described.

[0131] The term “comprising” or “comprises” where used herein means including the component(s) specified but not to the exclusion of the presence of other components. The term “consisting essentially of’ or “consists essentially of’ means including the components specified but excluding other components except for materials present as impurities, unavoidable materials present as a result of processes used to provide the components and the like.

[0132] The term “consisting of’ or “consists of’ means including the components specified but excluding other components.

[0133] Whenever appropriate, depending upon the context, the use of the term “comprises” or “comprising” may also be taken to include the meaning “consists essentially of’ or “consisting essentially of’, and also may also be taken to include the meaning “consists of’ or “consisting of’.

[0134] The optional features set out herein may be used either individually or in combination with each other where appropriate and particularly in the combinations as set out in the accompanying claims. The optional features for each aspect or exemplary embodiment of the invention, as set out herein are also applicable to all other aspects or exemplary embodiments of the invention, where appropriate. In other words, the skilled person reading this specification should consider the optional features for each aspect or exemplary embodiment of the invention as interchangeable and combinable between different aspects and exemplary embodiments.

[0135] It should be understood that while the use of words such as “preferable”, “preferably”, “preferred” or “more preferred” in the description suggest that a feature so described may be desirable, it may nevertheless not be necessary and embodiments lacking such a feature may be contemplated as within the scope of the invention as defined in the appended claims. In relation to the claims, it is intended that when words such as “a,” “an,” or “at least one,” are used to preface a feature there is no intention to limit the claim to only one such feature unless specifically stated to the contrary in the claim.

[0136] The invention is further described in the following non-limiting embodiments.

[0137] Numbered Embodiments

[0138] 1. A pharmaceutical composition comprising IL-15 or a derivative thereof, for use in combination with a chemotherapeutic agent in the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent.

[0139] 2. The pharmaceutical composition, for use according to embodiment 1 wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 minute and 1 month prior to said chemotherapeutic agent.

[0140] 3. The pharmaceutical composition, for use according to embodiment 1 or 2, wherein the IL-15 or derivative thereof is administered between 1 hour and 7 days prior to said chemotherapeutic agent.

[0141] 4. The pharmaceutical composition, for use according to any preceding embodiment wherein the IL-15 or derivative thereof comprises an IL-15 super agonist.

[0142] 5. The pharmaceutical composition, for use according to any preceding embodiment wherein the pharmaceutical composition comprises a complex of IL-15 and IL-15Ra.

[0143] 6. The pharmaceutical composition, for use according to any preceding embodiment wherein the pharmaceutical composition comprises a half-life extending moiety.

[0144] 7. The pharmaceutical composition, for use according to embodiment 6, wherein the half-life extending moiety is selected from an Fc domain, a modified Fc domain, PEG, serum albumin, a serum albumin binding molecule.

[0145] 8. The pharmaceutical composition, for use according to any preceding embodiment wherein the IL-15 or derivative thereof comprises a super agonist mutation.

[0146] 9. The pharmaceutical composition, for use according to embodiment 8, wherein the super agonist mutation is N72D (IL-15N72D).

[0147] 10. The pharmaceutical composition, for use according to any one of embodiments 5 to 10 wherein the IL-15Ra is provided as an IL-15RaSu / Fc fusion protein.

[0148] 11. The pharmaceutical composition, for use according to any preceding embodiment wherein the pharmaceutical composition comprises a dimeric IL-15RaSu / lgG1 Fc and two IL-15N72D molecules.

[0149] 12. The pharmaceutical composition, for use according to any preceding embodiment wherein the IL-15 or derivative thereof is selected from one or more of ALT803, XmAb306, NKTR-225, SO-C101 , NIZ985, NL-201 or rhlL-15.

[0150] 13. The pharmaceutical composition, for use according to any preceding embodiment, wherein the chemotherapeutic agent is selected from a platinum-based chemotherapeutic agent, an anthracycline-based chemotherapeutic agent, and alkylating chemotherapeutic agent, an taxane-based chemotherapeutic agent, a vinca alkaloid-based chemotherapeutic agent, a retinoid-based chemotherapeutic agent, a histone deacetylase inhibitor, a topoisomerase I inhibitor, a topoisomerase II inhibitor, a kinase inhibitor, or a nucleotide analogue-based chemotherapeutic agent.

[0151] 14. The pharmaceutical composition, for use according to any preceding embodiment, wherein the platinum-based chemotherapeutic agent is selected from carboplatin, cisplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin and / or combinations thereof. The pharmaceutical composition, for use according to any preceding embodiment wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered at a dose of between 0.1 pg / kg and 100 mg / kg. The pharmaceutical composition, for use according to any preceding embodiment wherein multiple doses of the pharmaceutical composition comprising IL-15 or derivative thereof are administered prior to said chemotherapeutic agent. The pharmaceutical composition, for use according to any preceding embodiment wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered systemically, intravenously, subcutaneously, intramuscularly, intravesically, or orally. The pharmaceutical composition, for use according to any preceding embodiment wherein the chemotherapeutic agent is administered systemically, intravenously, subcutaneously, intramuscularly, intravesically, intrathecally or orally. The pharmaceutical composition, for use according to any preceding embodiment, wherein the cancer is selected from ovarian cancer, pancreatic cancer, bone cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, breast cancer, brain cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the oesophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, kidney cancer, sarcoma of soft tissue, cancer of the urethra, cancer of the bladder, renal cancer, lung cancer, non-small cell lung cancer, thymoma, urothelial carcinoma leukaemia, prostate cancer, mesothelioma, adrenocortical carcinoma, lymphomas, such as such as Hodgkin's disease, nonHodgkin's, gastric cancer, and multiple myelomas. The pharmaceutical composition, for use according to any preceding embodiment, wherein a further anti-cancer agent is administered. The pharmaceutical composition, for use according to embodiment 20, wherein the further anti-cancer agent is selected from immunotherapy, hormone therapy, biologic, cytokine, small molecule, gene therapy, viral therapy, bone marrow transplantation, nanotherapy, targeted anti-cancer therapies, oncolytic drugs, CAR-T therapy, therapeutics targeting DNA damage repair and homologous recombination deficiency and / or autologous stem cell rescue (ASCR). The pharmaceutical composition, for use according to any preceding embodiment, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof increases the level of natural killer (NK) cells and / or CD8+ T cells. 23. The pharmaceutical composition, for use according to any preceding embodiment, wherein the subject received one or more further anti-cancer agent or anti-cancer treatment prior to the administration of the IL-15 or a derivative thereof.

[0152] 24. The pharmaceutical composition, for use according to any preceding embodiment, wherein the subject has increased expression, amplification of expression and / or mutation in the KRAS gene.

[0153] 25. The pharmaceutical composition, for use according to any preceding embodiment, wherein the subject has reduced expression, loss of expression and / or a mutation in the p53 gene.

[0154] 26. The pharmaceutical composition, for use according to any preceding embodiment, wherein the subject has reduced expression, loss of expression and / or a mutation in a Brea gene.

[0155] 27. The pharmaceutical composition, for use according to embodiment 26, wherein the Brea gene is the Brcal gene.

[0156] 28. A method of treating cancer in a subject, comprising administering a pharmaceutical composition comprising IL-15 or a derivative thereof, in combination with a chemotherapeutic agent to said subject, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the platinum-based chemotherapeutic agent.

[0157] 29. Use of a pharmaceutical composition comprising IL-15 or a derivative thereof, for the manufacture of a medicament for the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to a chemotherapeutic agent.

[0158] 30. An in vitro, ex vivo or in vivo method of sensitising cells to a chemotherapeutic agent comprising: contacting a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, contacting said population of cells to a chemotherapeutic agent.

[0159] 31 . An in vitro, ex vivo or in vivo method of enhancing efficacy of a chemotherapeutic agent comprising: exposing a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, exposing said population of cells to a chemotherapeutic agent.

[0160] 32. A method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for therapy.

[0161] 33. A method of therapy monitoring comprising: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, comparing the level of NK cells and / or CD8+ T cells with a reference value, determining the efficacy of the therapy based on the level of NK cells and / or CD8+ T cells compared to said reference value, wherein the subject has received, is currently receiving or will receive a therapy comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the IL-15 is administered prior to said chemotherapeutic agent.

[0162] 34. The method according to any one of embodiments 32 to 35, wherein the biological sample is selected from tissue, tumour, saliva, urine and / or blood including whole blood and / or plasma.

[0163] 35. The method according to embodiment 36, wherein the tissue is selected from pancreas, lung, omentum and / or spleen.

[0164] 36. The method according to any one of claims 32 to 36, wherein the NK cells comprise Nk1 , NK2, NK3 and / or NK4 cells, preferably NK3 and / or NK4.

[0165] 37. A method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises identifying the subject as having one of more of the following; increased expression, amplification of expression and / or mutation in the KRAS gene, reduced expression, loss of expression and / or a mutation in the p53 gene, reduced expression, loss of expression and / or a mutation in a Brea gene, and selecting said subject for therapy.

[0166] 38. A kit for the treatment of cancer comprising IL-15 or a derivative thereof and a chemotherapeutic agent.

[0167] EXAMPLES

[0168] Example 1

[0169] Platinum chemotherapeutics remain standard of care for many cancer patients, include high grade serous ovarian cancer following metastasis to the peritoneal cavity. It is becoming clear that chemotherapy can initiate adaptive immune responses to the cancer, however, it remains unclear to what extend innate Natural Killer (NK) cells contribute. Previous transcriptomic analysis of human ovarian cancer patient data suggests that NK cells are enriched after chemotherapy (1). Now, using two syngeneic models (UPK10 and ID8.p53- / -) of metastatic ovarian cancer we found that NK cells contribute towards the efficacy of carboplatin therapy in the UPK10 model. Besides investigating the mechanisms whereby NK cells mediate the effect of chemotherapy, we also asked if we could enhance chemotherapy in ovarian cancer by therapeutically expanding tissue-resident NK cells. To target NK cells, we used an IL-15 / IL-15Ra complex, which enhances the half-life and activity of IL-15.

[0170] IL-15 is a crucial cytokine for survival, proliferation and activation of NK and T cells (2). In complex with its soluble receptor IL-15Ra, it has been shown to increase NK cell and T cell numbers in vivo and induce their activation (3-5). Our data expands on and supports previous findings, showing that NK and T cells are expanded systemically (i.e. blood and spleen), but also within specific tissues including the omentum, the primary site of seeding for metastatic ovarian cancer (Figure 1 b and h). Within expanded NK cells, we show that specifically the more mature NK cell subtypes (NK3-NK4) are expanded (Figure 1g). NK and CD8 T cells are also more activated (Figure 1 l-m) and express higher levels of proliferation marker Ki67 (Figure 1 b, f, j-k). Cytotoxic cell types are also expanded in the pancreas and lung (data not shown), other potential tissues of interest for combination of IL-15 / IL-15Ra with conventional therapeutics.

[0171] Preliminary data suggested a negative impact of concurrent carboplatin treatment on immune cell expansion following IL-15 / IL-15Ra treatment (not shown). As IL-15 / IL-15Ra pushes cells into the cell cycle, it is hypothesised this makes them more sensitive to carboplatin cytotoxicity. Therefore, a sequential dosing regime was established where a 72-hour period was allowed between IL-15 / IL-15a and carboplatin treatment to allow for immune cell expansion (Figure 2a). Combination treated tumours were significantly smaller than carboplatin treated tumours (Figure 2b). Additionally, there was no significant difference in weight between combination treated UPK10 tumours and tumour naive omentum. Immunophenotyping of tumours shows that CD8 T cells, but not other immune subsets, are significantly enriched in the combination treated tumours, compared to untreated tumours (Figure 2c-g).

[0172] Preliminary data suggested a negative impact of concurrent carboplatin treatment on immune cell expansion following IL-15 / IL-15a treatment (not shown). As IL-15 / IL-15Ra pushes cells into the cell cycle, it is hypothesised this makes them more sensitive to carboplatin cytotoxicity. To optimise the therapeutic window for combination with carboplatin, a time course experiment was carried out (Figure 3a). Omental NK cells were significantly expanded following dosing, with numbers peaking 3 days post injection, before returning to baseline by day 14 (Figure 3b). One dose of IL-15SA also resulted in an expansion of total immune cells, which significantly peaked at day 3, as well as trends towards an increase in B cells, CD4 T cells and CD8 T cells. The expansion in NK cell numbers was driven by proliferation of tissue resident NK cells, as illustrated by the rapid induction of Ki67 expression 1 day after dosing (Figure 3c). One dose of IL-15SA also induced proliferation in CD8 T cells. Whilst NK cell population contracts post day 3, an enrichment of more mature, cytotoxic NK4 subsets is enriched at later timepoints (Figure 3d). Splenic NK cells were also significantly expanded following IL-15SA dosing, peaking 3 days post dosing, however the same trends were not observed in other lymphoid subsets (Figure 3e). Cytotoxic function of NK cells (Figure 2f) and CD8 T cells (Figure 3g) is also enhanced post treatment with IL-15SA. This peaks one day after dosing before returning to baseline. The percentage of Granzyme B positive cells remains elevated 3 days after treatment. Furthermore, one dose of IL-15SA was also sufficient to change CD8 T cell phenotypes, with an expansion of CD8 Memory cells, peaking at day 3 (Figure 3h). Importantly, expansion of CD8 T cells and NK cells was observed in the blood, also peaking at 3 days post dosing with IL-15SA (Figure 3i). Thereby suggesting immune cell numbers in the blood could be used as a readout response to treatment.

[0173] As cytotoxic immune cell number peaks 3 days post IL-15Ra dosing, a window of opportunity was identified in which to combine with carboplatin treatment. Therefore, a sequential dosing regime was established where a 72-hour period was given between IL-15 / IL-15a and carboplatin treatment to allow for immune cell expansion (Figure 4a). When harvested at a fixed endpoint, combination treated tumours were significantly smaller than carboplatin treated tumours (Figure 4b). Additionally, there was no significant difference in weight between combination treated UPK10 tumours and tumour naive omentum. Furthermore, in a survival study, there was a significant benefit of combination treatment compared to single treatments or control (Figure 4c). Finally, immunophenotyping of tumours harvested at day 24 highlighted that CD8 T cells, but not other immune subsets, are significantly enriched in the combination treated tumours, compared to untreated tumours (Figure 4d).

[0174] It is possible that the target patient population will have already received carboplatin treatment as standard of care. Therefore, to interrogate if sensitivity to IL-15SA induced immune expansion was impaired by prior treatment with chemotherapy, a new dosing regimen was tested where tumours were exposed first to two cycles of chemotherapy, followed by one dose of IL-15SA (Figure 5a). There was no significant difference in NK cell and CD8 T cell numbers between tumours treated with IL-15 only and tumours previously treated with carboplatin (Figure 5b).

[0175] Finally, UPK10 cells represent a genetic subset of ovarian cancer patients with amplifications and mutations in KRAS. To test if response to IL-15SA treatment in combination with chemotherapy could be applied to tumours with different genetics, a further murine mouse model of HGSOC was evaluated. ID8p53- / -Brca1- / - cell lines were injected IP into albino C57BI6 mice and bioluminescent tumour burden monitored weekly using in vivo imaging system (IVIS) until clinical endpoint. ID8p53- / -Brca1- / - tumour burden was significantly reduced in the IL-15SA + carboplatin combination (Combo) treatment group compared to carboplatin (CP) alone (Figure 6b), while overall survival was significantly longer in the combination treatment group compared to carboplatin alone (Figure 6c).

[0176] Thus, we are able to substantially enhance the efficacy of carboplatin in a model of ovarian cancer by expanding cytotoxic immune cell subsets, including NK cells, using the IL-15 / IL-15Ra compound.

[0177] Methods

[0178] Tumour models

[0179] Ovarian tumour models were established using murine syngeneic cell lines. UPK10 cells were provided by Conejo-Garcia, Moffit Cancer Centre1. For UPK10 cell lines, intraperitoneal injections of 1 e6 cells were performed into 8-12 week old C57BI / 6 mice. UPK10 tumours were harvested on day 24 post tumour cell injection. Mice were culled via cervical dislocation or CO2 overexposure.

[0180] ID8p53- / -BRCA1- / - were provided by McNeish, Imperial College. Cell lines were subsequently modified to express firefly luciferase. 4e6 ID8p53- / - BRCA1- / - cells were injected IP into 8-12- week-old albino C57 / BI6 mice. Bioluminescent imaging of tumour burden was carried out weekly using luciferin and the in vivo imaging system (IVIS). Mice where sch1 at clinical endpoint as determined by the onset of clinical signs including weight loss and onset of ascites.

[0181] Mice were culled via cervical dislocation or CO2 overexposure.

[0182] Treatment

[0183] Treatment of intraperitoneal tumours with dPBS, 250 ptg carboplatin (Fresenius-Kabi, #PL18727 / 0025) or IL-15 / IL-15Ra was performed by intraperitoneal injection of tumour naive or tumour bearing C57BI.6 mice. IL-15 / IL-15Ra complexes were forms by incubating 0.5|j.g IL-15 (Peprotech, #210-15) and 3|u.g IL-15Ra (R+D, #551-MR-100) at 37oC for 30 minutes prior to injection.

[0184] Tissue processing

[0185] For flow cytometry, tissues were processed as follows.

[0186] Spleens were homogenised using the back of a 5 ml syringe and a 70 |j.M cell strainer, repeatedly washed in PBS and transferred to a 15 ml conical tube. Samples were centrifuged (5 minutes, 1500 rpm) and supernatant removed. Red blood cells were lysed using (Biolegend, #420301) as per the manufacturer's instructions. Samples were resuspended in FACS buffer prior to staining. Omentums were collected in 500|LII HBSS (Thermo, #24020091). Soybean trypsin inhibitor and digest mixture (Collagenase I and DNAse I) were added to make a 1x final digest concentration. Samples were then incubated at 37°C, 10OOrpm for 45 minutes. Subsequently, omentums were homogenised with a 2ml syringe over a 70 |j.M cell strainer. Filter was washed with 10ml RPMI containing 10 % FBS. Samples were resuspended in FACS buffer prior to staining.

[0187] Tumours were collected in 500j_tl HBSS. Tumours were removed from collection media, roughly chopped using a scalpel and returned to collection media. Soybean trypsin inhibitor and digest mixture (Collagenase I and DNAse I) were added to make a 1x final digest concentration. Samples were then incubated at 37°C, OOrpm for 45 minutes. Subsequently, tumours were homogenised with a 2ml syringe over a 70 |j.M cell strainer. Filter was washed with 10ml RPMI containing 10 % FBS. If required, red blood cells were lysed using red blood cell lysis buffer (Biolegend, #420301) as per the manufacturer's instructions. Samples were resuspended in FACS buffer prior to staining.

[0188] Flow cytometry staining

[0189] Samples to be stained were resuspended in FACS buffer (PBS containing 1 % FBS and 2mM EDTA). Samples were stained in a 96-well U-bottom plate. Samples were centrifuged (3 minutes, 1500rpm) and resuspend in TruStain FcX™ (anti-mouse CD16 / 32)(Biolegend, #101319) for 10 minutes, on ice. Antibodies are diluted in FACS buffer and extracellular staining performed in 50 j L volume for 30 minutes at 4 °C in the dark. 100 j L FACS buffer was added to samples, centrifuge the plate and wash again with 150 j L FACS buffer. Prior to intracellular staining, cells are fixed and permeabilised using the Foxp3 fixation / permeabilisation kit (eBiosciences #00-5523-00) as per manufacturer’s instructions. Subsequently, intracellular antibodies are diluted in permeabilisation buffer and staining performed overnight at 4 °C. Samples were washed twice with permeabilisation buffer prior to resuspension in 200 j L FACS buffer. Samples were stored for < 48 hours prior to acquisition. Samples were acquired using Cytek Aurora.

[0190] If an activation panel is required cells are restimulated as follows. Unstained samples to be restimulated are resuspended in 200ul of RPMI (Thermo, #21875034) with 10% FBS (Thermo, #10270106) containing Cell Stimulation Cocktail (plus protein transport inhibitors) (Thermo, #00- 4975-93) or Protein Transport Inhibitor Cocktail alone (Thermo, #00-4980-93). Samples were incubated for 2 hours at 37oC prior to following the standard staining protocol.

[0191] Total cell type count was calculated as follows for each cell type:

[0192] Flow cytometry panels

[0193]

[0194] References

[0195] 1. Jimenez-Sanchez, A. et al. Unraveling tumor-immune heterogeneity in advanced ovarian cancer uncovers immunogenic effect of chemotherapy. Nature Genetics 52, 582-593 (2020).

[0196] 2. Ma, S., Caligiuri, M. A. & Yu, J. Harnessing IL-15 signaling to potentiate NK cell-mediated cancer immunotherapy. Trends in Immunology 43, 833-847 (2022).

[0197] 3. Dubois, S., Patel, H. J., Zhang, M., Waldmann, T. A. & Muller, J. R. Preassociation of IL-15 with IL-15R alpha-lgG1-Fc enhances its activity on proliferation of NK and CD8+ / CD44high T cells and its antitumor action. J Immunol 180, 2099-2106 (2008).

[0198] 4. Bessard, A., Sole, V., Bouchaud, G., Quemener, A. & Jacques, Y. High antitumor activity of RLI, an interleukin-15 (IL-15)-IL-15 receptor alpha fusion protein, in metastatic melanoma and colorectal cancer. Mol Cancer Ther 8, 2736-2745 (2009).

[0199] 5. Stoklasek, T. A., Schluns, K. S. & Lefrangois, L. Combined IL-15 / IL-15Ra Immunotherapy Maximizes IL-15 Activity In Vivo. J Immunol 177, 6072-6080 (2006).

Claims

CLAIMS1. A pharmaceutical composition comprising IL-15 or a derivative thereof, for use in combination with a chemotherapeutic agent in the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the chemotherapeutic agent.

2. The pharmaceutical composition, for use according to claim 1 wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered between 1 minute and 1 month prior to said chemotherapeutic agent.

3. The pharmaceutical composition, for use according to claim 1 or 2, wherein the IL-15 or derivative thereof is administered between 1 hour and 7 days prior to said chemotherapeutic agent.

4. The pharmaceutical composition, for use according to any preceding claim wherein the IL-15 or derivative thereof comprises an IL-15 super agonist.

5. The pharmaceutical composition, for use according to any preceding claim wherein the pharmaceutical composition comprises a complex of IL-15 and IL-15Ra.

6. The pharmaceutical composition, for use according to any preceding claim wherein the pharmaceutical composition comprises a half-life extending moiety.

7. The pharmaceutical composition, for use according to claim 6, wherein the half-life extending moiety is selected from an Fc domain, a modified Fc domain, PEG, serum albumin, a serum albumin binding molecule.

8. The pharmaceutical composition, for use according to any preceding claim wherein the IL-15 or derivative thereof comprises a super agonist mutation.

9. The pharmaceutical composition, for use according to claim 8, wherein the super agonist mutation is N72D (IL-15N72D).

10. The pharmaceutical composition, for use according to any one of claims 5 to 10 wherein the IL-15Ra is provided as an IL-15RaSu / Fc fusion protein.

11. The pharmaceutical composition, for use according to any preceding claim wherein the pharmaceutical composition comprises a dimeric IL-15RaSu / lgG1 Fc and two IL- 15N72D molecules.

12. The pharmaceutical composition, for use according to any preceding claim wherein the IL-15 or derivative thereof is selected from one or more of ALT803, XmAb306, NKTR-225, SO-C101 , NIZ985, NL-201 or rhlL-15.

13. The pharmaceutical composition, for use according to any preceding claim, wherein the chemotherapeutic agent is selected from a platinum-based chemotherapeutic agent, an anthracycline-based chemotherapeutic agent, and alkylating chemotherapeutic agent, an taxane-based chemotherapeutic agent, a vinca alkaloid- based chemotherapeutic agent, a retinoid-based chemotherapeutic agent, a histonedeacetylase inhibitor, a topoisomerase I inhibitor, a topoisomerase II inhibitor, a kinase inhibitor, or a nucleotide analogue-based chemotherapeutic agent.

14. The pharmaceutical composition, for use according to any preceding claim, wherein the platinum-based chemotherapeutic agent is selected from carboplatin, cisplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin and / or combinations thereof.

15. The pharmaceutical composition, for use according to any preceding claim wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered at a dose of between 0.1 pg / kg and 100 mg / kg.

16. The pharmaceutical composition, for use according to any preceding claim wherein multiple doses of the pharmaceutical composition comprising IL-15 or derivative thereof are administered prior to said chemotherapeutic agent.

17. The pharmaceutical composition, for use according to any preceding claim wherein the pharmaceutical composition comprising IL-15 or derivative thereof is administered systemically, intravenously, subcutaneously, intramuscularly, intravesically, or orally.

18. The pharmaceutical composition, for use according to any preceding claim wherein the chemotherapeutic agent is administered systemically, intravenously, subcutaneously, intramuscularly, intravesically, intrathecally or orally.

19. The pharmaceutical composition, for use according to any preceding claim, wherein the cancer is selected from ovarian cancer, pancreatic cancer, bone cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, breast cancer, brain cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the oesophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, kidney cancer, sarcoma of soft tissue, cancer of the urethra, cancer of the bladder, renal cancer, lung cancer, non-small cell lung cancer, thymoma, urothelial carcinoma leukaemia, prostate cancer, mesothelioma, adrenocortical carcinoma, lymphomas, such as such as Hodgkin's disease, nonHodgkin's, gastric cancer, and multiple myelomas.

20. The pharmaceutical composition, for use according to any preceding claim, wherein a further anti-cancer agent is administered.

21. The pharmaceutical composition, for use according to claim 20, wherein the further anti-cancer agent is selected from immunotherapy, hormone therapy, biologic, cytokine, small molecule, gene therapy, viral therapy, bone marrow transplantation, nanotherapy, targeted anti-cancer therapies, oncolytic drugs, CAR-T therapy, therapeutics targeting DNA damage repair and homologous recombination deficiency and / or autologous stem cell rescue (ASCR).

22. The pharmaceutical composition, for use according to any preceding claim, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof increases the level of natural killer (NK) cells and / or CD8+ T cells.

23. The pharmaceutical composition, for use according to any preceding claim, wherein the subject received one or more further anti-cancer agent or anti-cancer treatment prior to the administration of the IL-15 or a derivative thereof.

24. The pharmaceutical composition, for use according to any preceding claim, wherein the subject has increased expression, amplification of expression and / or mutation in the KRAS gene.

25. The pharmaceutical composition, for use according to any preceding claim, wherein the subject has reduced expression, loss of expression and / or a mutation in the p53 gene.

26. The pharmaceutical composition, for use according to any preceding claim, wherein the subject has reduced expression, loss of expression and / or a mutation in a Brea gene.

27. The pharmaceutical composition, for use according to claim 26, wherein the Brea gene is the Brcal gene.

28. A method of treating cancer in a subject, comprising administering a pharmaceutical composition comprising IL-15 or a derivative thereof, in combination with a chemotherapeutic agent to said subject, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to the platinum-based chemotherapeutic agent.

29. Use of a pharmaceutical composition comprising IL-15 or a derivative thereof, for the manufacture of a medicament for the treatment of cancer, wherein the pharmaceutical composition comprising IL-15 or a derivative thereof is administered prior to a chemotherapeutic agent.

30. An in vitro, ex vivo or in vivo method of sensitising cells to a chemotherapeutic agent comprising: contacting a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, contacting said population of cells to a chemotherapeutic agent.31 . An in vitro, ex vivo or in vivo method of enhancing efficacy of a chemotherapeutic agent comprising: exposing a population of cells to a pharmaceutical composition comprising IL-15 or a derivative thereof, exposing said population of cells to a chemotherapeutic agent.

32. A method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent,wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, identifying said subject as having above a threshold level of NK cells and / or CD8+ T cells present, and selecting said subject for therapy.

33. A method of therapy monitoring comprising: screening a biological sample obtained from a subject with cancer for the presence of NK cells and / or CD8+ T cells, comparing the level of NK cells and / or CD8+ T cells with a reference value, determining the efficacy of the therapy based on the level of NK cells and / or CD8+ T cells compared to said reference value, wherein the subject has received, is currently receiving or will receive a therapy comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the IL-15 is administered prior to said chemotherapeutic agent.

34. The method according to any one of claims 32 to 33, wherein the biological sample is selected from tissue, tumour, saliva, urine and / or blood including whole blood and / or plasma.

35. The method according to claim 34, wherein the tissue is selected from pancreas, lung, omentum and / or spleen.

36. The method according to any one of claims 32 to 36, wherein the NK cells comprise Nk1 , NK2, NK3 and / or NK4 cells, preferably NK3 and / or NK4.

37. A method of identifying a subject as suitable for therapy with a pharmaceutical composition comprising IL-15 or a derivative thereof and a chemotherapeutic agent, wherein the pharmaceutical composition is administered prior to said chemotherapeutic agent, wherein the method comprises identifying the subject as having one of more of the following; increased expression, amplification of expression and / or mutation in the KRAS gene, reduced expression, loss of expression and / or a mutation in the p53 gene, reduced expression, loss of expression and / or a mutation in a Brea gene, and selecting said subject for therapy38. A kit for the treatment of cancer comprising IL-15 or a derivative thereof and a chemotherapeutic agent and optionally instructions for use.

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