TECHNICAL FIELD

The present invention relates to a WNT5A peptide for use in neoadjuvant therapy in a cancer patient to reduce the patient's TNM disease stage.

BACKGROUND OF THE INVENTION

In most cancers, disease progression starts with the formation of a tumour, also referred to as a primary tumour. The tumour may be benign, i.e. not cancerous, or malignant, i.e. cancerous. The benign tumour may grow large but does not propagation into, or invade, nearby tissues or other parts of the body. A malignant tumour, on the other hand, can both grow and metastasize, that is propagation into, or invade, nearby tissues and organs. Furthermore, the tumours can propagate by metastasis to other parts of the body through the blood and lymph systems. When cancer cells metastasize and form secondary tumours, the cells in the secondary tumour may be like those in the original primary tumour.

Given the significant role of the primary tumour in the progression of a cancer, it is essential that the tumour is treated efficiently in order to improve the overall outcome for a patient. Once a tumour has been diagnosed as a cancer, the typical course of treatment involves the surgical removal of the primary tumour followed by, and depending on the cancer type, various treatment regimes, including radiation, administration of chemotherapeutic drugs, endocrine treatment or other novel biological treatments.

There may exist situations in which the tumour cannot be removed completely or can only be removed partially through surgery. Such tumours may be referred to as unresectable tumours. Examples of such situations include when the tumour is too large to be safely removed through surgery, the tumour is in an essential organ and removing it would mean removing too much of the organ, or when the tumour is intertwined with blood vessels or other vital structures in the body, such as in the case of brain cancer, and removing such cancer could pose too much of a risk to the patient. In such cases neoadjuvant treatment of the cancer may in some cases be offered before surgery with the goal of reducing the patient's disease stage as defined by the tumour, node, metastasis (TNM) stage classification system (see e.g. the AJCC Cancer Staging Manual, Eighth Edition (2017), published by Springer International Publishing), also referred to herein as "downstaging", before resective surgery is performed. The result of downstaging typically includes shrinkage of the tumour and/or stopping the propagation or invasive behaviour of the cancer cells, e.g. to lymph nodes, to make surgery less extensive and more effective. However, current neoadjuvant therapies are limited to radiopharmaceuticals, radiation, targeted therapy drugs, immunotherapy medications, chemotherapeutics or hormone therapy drugs which therapies are often associated with undesirable side effects and/or a less than optimal downstaging effect on the cancer.

On this background it is therefore an object of the present invention to provide improved neoadjuvant therapies for use in the treatment of primary cancer tumours which therapies entail fewer side effects and comprise administering a therapeutic agent that is better tolerated by the patients.

SUMMARY OF THE INVENTION

The Wnt (Wingless-related integration site) protein family contains highly conserved proteins that play a role in embryonic development such as body axis patterning, cell proliferation and migration. Genetic mutation in the Wnt signalling pathway may cause breast cancer, prostate cancer glioblastoma, type II diabetes and other diseases. WNT5A (Wnt Family Member 5A), an autocrine and paracrine -catenin independent ligand, has been shown to either suppress or promote cancer progression, depending on the type of cancer. In patients with colorectal cancer, WNT5A triggers a variety of downstream signalling pathways that mainly suppress the migration and invasion of cancer cells.

It has now surprisingly been found that a WNT5A peptide derived from the amino acid sequence of the WNT5A molecule and selected from the group consisting of:

MDGCEL (SEQ. ID. NO. 3),

GMDGCEL (SEQ. ID. NO. 4),

EGMDGCEL (SEQ. ID. NO. 5),

SEGMDGCEL (SEQ. ID. NO. 6),

TSEGMDGCEL (SEQ. ID. NO. 7),

KTSEGMDGCEL (SEQ. ID. NO. 8),

NKTSEGMDGCEL (SEQ. ID. NO. 9),

CNKTSEGMDGCEL (SEQ. ID. NO. 10),

LCNKTSEGMDGCEL (SEQ. ID. NO. 11),

RLCNKTSEGMDGCEL (SEQ. ID. NO. 12),

GRLCNKTSEGMDGCEL (SEQ. ID. NO. 13), QGRLCNKTSEGMDGCEL (SEQ. ID. NO. 14), TQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 15), GTQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 16), and LGTQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 17), or a formylated derivative thereof, is effective in reducing tumour growth i.e. its propagation and/or volume in a patient diagnosed with a cancer. This has been shown both through in vivo animal studies, see Fig. 3 - 4. (Osman et al, Anticancer Research 39:1719-1728, 2019), and in the Phase l/ll NeoFox clinical study in human patients with stage II / III colon cancer, who at diagnosis are considered to have a high risk of recurrence after the primary tumour has been surgically resected, see Fig. 1 - 2.

The evaluation of tumour growth i.e. its propagation and/or volume is typically made on the basis of X-rays and scans of tumours in order to estimate their size and propagation, such as by computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET)/CT, and ultrasound scans. By comparing scans obtained over time, the medical professional can see how a tumour is growing and propagating and in that way for example determine how the cancer is responding to treatment.

In a first aspect of the invention, there is provided a WNT5A peptide for neoadjuvant (preoperative) use in the reduction or elimination of tumour growth i.e. its propagation and/or volume in a patient diagnosed with a primary cancer.

In the context of the first aspect, a patient diagnosed with a cancer and having the tumour is to be understood as a human of any age suffering from a cancer which is associated with the formation of one or more tumours in the body. The patient will have been recently diagnosed with cancer and has been scheduled for resective surgery. According to the first aspect, the patient's initial TNM status is determined, whereafter a WNT5A peptide of the present invention is administered to said patient before surgery with the goal of reducing the patient's disease stage as defined by the tumour, node, metastasis (TNM) stage classification system (8 ^th edition, 2017), also referred to herein as "downstaging", before resective surgery is performed. Downstaging is performed to make surgery less extensive and more effective and to impair the progression of the cancer disease.

For all aspects and embodiments of the present invention, the WNT5A peptide comprises amino acid sequence XDGXEL (SEQ. ID. NO. 2), or a formylated derivative thereof, wherein X in position 1 is methionine (M) or norleucine (Nle) and X in position 4 is cysteine (C) or alanine (A). BRIEF DESCRIPTION OF THE FIGURES

The invention will be described in more detail below by means of non-limiting examples of embodiments and with reference to the figures, in which:

Figures 1 and 2 show the TNM downstaging effect in human patients of Foxy-5 treatment vs. Control from the NeoFox clinical study. As can be seen, downstaging is significantly more frequent in patients receiving neoadjuvant Foxy-5 treatment than in patients receiving standard of care treatment.

Figure 3 shows the in vivo effect of Foxy-5 treatment on active p-catenin nuclei expression in HT-29 colon cancer tissue as visualized by immunohistochemistry (IHC) (Osman et al). The IHC stainings were scored by having representative photos from saline- and Foxy-5-treated animals presented. From each such slide, 4 boxes were put on top of the stained tissue and for each of them the p-catenin staining was scored as a percentage of stained cells multiplied by the staining intensity. The following score was used for percentage stained cells; 0 for positive stained cells <5%, 1 for positive cells 5-25%, 2 for positive cells 26-50%, 3 for positive cells 51-75%, 4 for positive cells >75%. For staining intensities, the stained areas were scored as; 1 for weak staining, 2 for medium staining and 3 for strong staining. For each box a final score was obtained by multiplying the score for percentage of positively stained cells with the score for staining intensity. Finally, a mean value was obtained from the 4 boxes for each tumour. The same scoring protocol was then used for all IHC analyses presented in the present application.

Figure 4 shows the in vivo effect of Foxy-5 treatment on active p-catenin nuclei expression in Caco-2 colon cancer tissue as visualized by IHC (Osman et al). The IHC staining were scored exactly as described in detail for the IHC staining in Figure 4. In the lower right panel, the size of the tumours for both the saline- and Foxy- 5-treated animal tumour are presented.

Figures 3 and 4 further show the in vivo effect of Foxy-5 treatment on tumour volume in HT-29 and Caco-2 colon cancer xenograph tissue, respectively. The figures outline the effects of Foxy-5 treatment on p-catenin signalling and tumour volume of HT29 and Caco-2 derived tumours compared to that from control (vehicle- treated) mice. DEFINITIONS

In the context of the present inventive concept, "proliferation" is used interchangeably with growth. The term "growth" also covers the situation where a primary tumour invades neighbouring tissue or organs.

In the context of the present invention, a primary tumour is the original, or first cancer tissue in the body of the patient diagnosed with cancer.

The term "signalling properties", as used herein, means binding of the WNT5A or the Foxy-5 peptide to primarily a Frizzled receptor protein (Fz) followed by an intracellular signalling cascade in the cell eventually leading to reduction or elimination of tumour growth and propagation.

In the context of the present invention, a "WNT5A agonist" means a WNT5A-derived agonistic peptide which is capable of mimicking the function of WNT5A, thus reconstituting WNT5A-signalling in cancer tissue that lack an endogenous expression of WNT5A.

In the context of the present invention, "downstaging" shall refer to a reduction of the patient's tumour load, i.e. in the TNM stage of a cancer, from a more to a less threatening stage as defined by the tumour, node, metastasis (TNM) stage classification system (as described in AJCC Cancer Staging Manual, Eighth Edition (2017), published by Springer International Publishing) which includes assessment of the size and growth of a tumour, and/or its invasion and propagation into the primary and adjacent structures and organs, the extent of metastasis, and/or the lymph node involvement.

DETAILED DESCRIPTION OF THE INVENTION

WNT5A is a 44-50 kDa member of the Wnt family of proteins which is expressed by many normal cells in the body. WNT5A is secreted from the cells and exerts its action on the same or neighbouring cells by binding to and activating a receptor complex primarily involving a Frizzled receptor. The WNT5A protein is known to activate a receptor called Frizzled 5. Upon activation of the Frizzled 5 receptor a series of signalling events inside of the cells are activated, where one of the first events is the generation of a short-lived increase in calcium levels inside of the cell, a so-called calcium-signal. The calcium-signal in turn triggers a series of further signalling events leading to a change in the functions of the cells, such as adhesion and migration. Thus, activating a Frizzled receptor leads to signalling events inside the cell, resulting in increased adherence of the cell to its neighbouring cells and its adhesion to the surrounding connective tissue resulting in decreased ability of a cancer cell to migrate to structures in the vicinity, such as lymph nodes and blood vessels. In healthy breast epithelial cells for example, WNT5A is highly expressed and secures a firm adherence between cells and to the surrounding basement membrane and thereby restricts migration of the cells.

Some cancers/cancer cells have been found to be associated with a low endogenous expression of WNT5A. As a relatively large protein WNT5A itself is not a straightforward drug candidate, but the present inventors have theorized that a WNT5A agonistic peptide may reconstitute WNT5A signalling in cancer cells which lack an endogenous expression of WNT5A.

As mentioned hereinabove, it has surprisingly been found that a small (< 20 amino acids) peptide derived from the amino acid sequence of the WNT5A molecule and selected from the group consisting of:

MDGCEL (SEQ. ID. NO. 3), GMDGCEL (SEQ. ID. NO. 4), EGMDGCEL (SEQ. ID. NO. 5), SEGMDGCEL (SEQ. ID. NO. 6), TSEGMDGCEL (SEQ. ID. NO. 7), KTSEGMDGCEL (SEQ. ID. NO. 8), NKTSEGMDGCEL (SEQ. ID. NO. 9), CNKTSEGMDGCEL (SEQ. ID. NO. 10), LCNKTSEGMDGCEL (SEQ. ID. NO. 11), RLCNKTSEGMDGCEL (SEQ. ID. NO. 12), GRLCNKTSEGMDGCEL (SEQ. ID. NO. 13), QGRLCNKTSEGMDGCEL (SEQ. ID. NO. 14),

TQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 15),

GTQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 16), and

LGTQGRLCNKTSEGMDGCEL (SEQ. ID. NO. 17), or a formylated derivative thereof, is effective in reducing tumour growth, i.e. its propagation and/or volume, in a patient diagnosed with a cancer. Such peptides are referred to hereinafter as "WNT5A peptides".

The effect of the WNT5A peptides of the invention on cancer cells has been shown both through in vivo animal studies, see Fig 3-4, and in the Phase l/ll NeoFox clinical study in human patients with stage II / III colon cancer, who at diagnosis are considered to have a high risk of recurrence after the primary tumour has been surgically resected, see Fig 1-2. The inventors have thus found that neoadjuvant treatment of cancer patients with Foxy-5, the WNT5A peptide agonist N-Formyl-Met-Asp-Gly-Cys-Glu-Leu-OH, leads to a statistically significant downstaging in TNM status (tumour, nodes and metastasis), i.e. a reduction of TNM stage, during the time period between diagnosis and surgery when compared to a control group not treated with Foxy-5.

In a first aspect of the invention, there is thus provided a WNT5A peptide for neoadjuvant (preoperative) use in the reduction or elimination of tumour growth i.e. its propagation and/or volume in a patient diagnosed with a cancer and having said tumour. In the context of the first aspect, a patient diagnosed with a cancer and having the tumour is to be understood as a human of any age suffering from a cancer which is associated with the formation of one or more tumours in the body.

In an embodiment of the first aspect the patient will have been recently diagnosed with cancer for the first time or diagnosed with cancer again and has been scheduled for resective surgery. According to the first aspect, the patient's initial TNM status is determined, e.g. in connection with said diagnosis being made, whereafter a WNT5A peptide of the present invention is administered to said patient before surgery with the goal of reducing the patient's disease stage as defined by the tumour, node, metastasis (TNM) stage classification system (8 ^th edition, 2017), also referred to herein as "downstaging", before resective surgery is performed. A downstaging effect is thus observed if the patient's TNM status is lower at the time of resective surgery (the preoperative TNM status) than at the time the initial TNM status was determined. The preoperative TNM status can be determined in the same way as the initial TNM status, i.e. typically by CT/MRI scanning. However, the patient's preoperative TNM status can also be determined after resective surgery has taken place, e.g. based on observations made during surgery or based on the final pathology report prepared after resective surgery has been completed. A patient's post-operative TNM status may also be evaluated in the same way as the initial TNM status, i.e. typically by CT/MRI scanning.

In an embodiment of the first aspect an effective dose of the WNT5A peptide is administered to said patient after said diagnosis has been made. In another embodiment of the first aspect an effective dose of the WNT5A peptide is administered to said patient right after said diagnosis has been made and resective surgery is planned for.

In yet another embodiment of the first aspect, the WNT5A peptide is administered repeatedly to said patient until the point where surgery may be contemplated. In yet another embodiment, the WNT5A peptide is administered as at least one dose of between 1 - 10 mg/kg body weight, such as between 3 - 9 mg/kg, such as between 4 - 8 mg/kg, such as 6 mg/kg body weight over a period of 1 - 6 weeks prior to surgery, such as 2 - 5 weeks, such as 3 weeks prior to surgery.

In a preferred embodiment of the first aspect, the patient receives at least twelve (12) administrations of the WNT5A peptide as monotherapy (1.8 up to 8 mg/kg, intravenously [IV], three to five administrations per week) prior to planned surgery.

In an embodiment of the first aspect the patient's preoperative TNM status is determined before, such as immediately before resective surgery, for example by CT/MRI scanning. In another embodiment the patient's preoperative TNM status is determined during or at any time after surgery has taken place. In another embodiment any removed tumour tissue and observations of regional lymph nodes, adjacent structures and organs made during surgery and/or the observations made in a pathology report are considered when determining the preoperative TNM status. For all embodiments herein, downstaging is determined based on the difference in the patient's initial TNM status compared to the patient's preoperative TNM status regardless of how the preoperative TNM status has been determined.

In one embodiment the patient's preoperative TNM status is not so low that resective surgery is recommended, in which case the administration of the WNT5A peptide to said patient is continued until a new assessment of the patient's preoperative TNM status indicates that resective surgery may be contemplated.

In an embodiment the patient's postoperative TNM status may be determined at any time after surgery, preferably by CT/MRI scanning, providing a medical professional an opportunity to make a new prognosis or diagnosis.

In one embodiment, the downstaging is a result of a reduction or elimination of tumour growth and/or propagation of a primary tumour in the patient. In another embodiment, the downstaging is a result of a reduction or elimination of a primary tumour's invasion into adjacent structures and organs. In another embodiment, the downstaging is a result of a reduction of affected regional lymph nodes, i.e. when cancer cells have propagation to the lymph nodes from a primary cancer located somewhere else in the body. Finally, in yet another embodiment, downstaging is a result of a reduction or elimination of metastases originating from the original primary tumour.

In a preferred embodiment, downstaging is a result of administering a WNT5A peptide of the present invention to the patient over the period between initial diagnosis and resective surgery. In another embodiment, the downstaging has amounted to a reduction from TNM stage III to a lower stage, such as from TNM Stage Illa to Stage lib or lie or lower. In another embodiment, the downstaging has amounted to a reduction of TNM stage II to a lower stage, such as from TNM stage He or lib to a lower stage, such as from TNM Stage He to lib or lower, or from TNM Stage lib to Ila or lower.

In a preferred embodiment there is provided a WNT5A peptide for use in downstaging of cancer tumour tissue in a patient, wherein the cancer is selected from the list consisting of prostate cancer, breast cancer, colon cancer, colorectal cancer, ovarian cancer, thyroid cancer, and liver cancer, and wherein said downstaging amounts to a reduction over the period between initial diagnosis and resective surgery from TNM stage III to a lower stage, such as from TNM Stage Illa to Stage lib or lie or lower. In another embodiment, the downstaging has amounted to a reduction of TNM stage II to a lower stage, such as from TNM stage lie or lib to a lower stage, such as from TNM Stage lie to lib or lower, or from TNM Stage lib to Ila or lower. In a further embodiment, the cancer is colon cancer or breast cancer.

According to separate embodiments of the present invention, there is provided a WNT5A peptide for use in the reduction or elimination of tumour growth, i.e. its propagation and/or volume in a patient diagnosed with a cancer, wherein the cancer is selected from the list consisting of prostate cancer, breast cancer, colon cancer, colorectal cancer, ovarian cancer, thyroid cancer, and liver cancer. In a further embodiment, the cancer is colon cancer or breast cancer. In another embodiment the cancer is an invasive (or infiltrating) adenocarcinoma.

In one preferred embodiment, the WNT5A peptide is a WNT5A agonist. As mentioned, the full length WNT5A protein has several disadvantages as drug candidate. Being a large protein (44-50 kDa), the WNT5A protein requires a complex and lengthy synthesis just in order to produce the correct primary amino acid sequence and post-translational modifications, and in addition WNT5A has a heparan sulphate-binding domain which may limit its distribution in the body. Therefore, a smaller peptide that mimics the effect of WNT5A is more preferred.

In an embodiment, the total length of the WNT5A peptide is equal to or less than 50 amino acids, such as equal to or less than 20 amino acids. WNT5A peptides of the present invention comprising the amino acid sequence XDGXEL (SEQ ID NO. 2) also comprises WNT5A mimicking effects. The WNT5A peptides of varying lengths and comprising the sequence XDGXEL (SEQ ID NO: 2) may be produced synthetically, for example by liquid phase or solid phase peptide synthesis. The peptide of varying lengths has 20 amino acids or less, more preferred 10 amino acids or less. Preferably, the WNT5A peptide is a hexapeptide consisting of 6 amino acids. In one embodiment, the amino acid sequence of the WNT5A peptide is XDGXEL, wherein the X in position 1 is methionine (M) or norleucine and X in position 4 is cysteine (C) or alanine (A).

In one embodiment, the WNT5A peptide is MDGCEL (SEQ. ID. NO. 3). In a preferred embodiment methionine in position 1 is derivatized as formylated methionine (N-formyl methionine). This formylated hexapeptide is denoted Foxy-5, i.e. N-Formyl-Met-Asp-Gly-Cys-Glu-Leu-OH. The modification/derivatization (formylation) of one amino acid improves the effect of the peptide and makes it more effective and resistant to degradation in vivo.

In yet another embodiment of the first aspect the neoadjuvant (preoperative) treatment of said patient with a WNT5A peptide according to the invention comprises administering an effective amount of the WNT5A peptide immediately after diagnosis of the cancer. In a preferred embodiment, the WNT5A peptide is administered repeatedly until resective surgery is contemplated, such as three to five administrations per week prior to planned surgery. Preferably at least twelve (12) administrations of the WNT5A peptide are given prior to planned surgery.

Adjuvant administration of a WNT5A peptide according to the invention may be continued post- operatively but is not mandatory.

For all aspects and embodiments of the present invention, the WNT5A peptide comprises amino acid sequence XDGXEL (SEQ. ID. NO. 2), or a formylated derivative thereof, wherein X in position 1 is methionine (M) or norleucine (Nle) and X in position 4 is cysteine (C) or alanine (A). In a preferred embodiment of all aspects and embodiments of the present invention the WNT5A peptide is Foxy-5, which has been demonstrated to be a true WNT5A agonist since it triggers the same signalling events and functional responses as WNT5A. For neoadjuvant treatment purposes Foxy-5 is preferably administered intravenously (IV) as monotherapy. EXAMPLES

Example 1

5 The NeoFox phase 2 study with Foxy-5 includes human patients with stage II / III colon cancer and consists of two parts. The part 1 is the original design where 127 patients, who at diagnosis are considered to have a high risk of recurrence after the primary tumour has been surgically resected, were randomized. Ad hoc observations were made in this patient population and led to a revision of the study design to confirm the observation. The study has thus been amended to include part 2, where 80 additional patients will be0 included.

The aim of the amended study is to evaluate whether Foxy-5 reduces the size of the local propagation in the intestine in patients with stage III colon cancer, i.e. that the primary tumor's invasion of the intestinal wall and to local lymph nodes decreases. Any effect of Foxy-5 on primary tumour and lymph nodes will be studied radiologically using CT/MRI. The evaluation involves a comparison of the effect of Foxy-5 against a control5 group without Foxy-5 treatment. In addition, a pathological examination of surgically removed tumors and lymph nodes will be performed using microscopy.

The study is currently ongoing at approximately 25 hospitals in Spain and Hungary.

The dose administration with Foxy-5 begins in connection with the diagnosis and continues until surgery, when it is terminated, and the effect will be evaluated. This means that subjects are treated for at least 30 weeks. The patient normally receives surgery within 4 weeks from the time of diagnosis.

EVALUATION TO SEE REDUCED SIZE OF LOCAL SPREAD

Patients in part 2 of the NeoFox study are randomized 1:1 to either the Foxy-5 Treatment Arm (up to 40 subjects) or Control Arm (up to 40 subjects). All study subjects who are randomized will be treated either5 with Foxy-5 or serve as control and will be evaluated for safety and efficacy.

The evaluation in NeoFox is a comparison of the safety and efficacy of Foxy-5 compared to a control group without Foxy-5 treatment. Based on CT/MRI the subjects should be judged to be

T3 or T4, N1 or N2, and M0 as per TNM (Tumor, Node, Metastasis) classification of colon cancer (8 ^th edition, 2017) in order to be0 included in the study. All subjects randomized to the Foxy-5 Arm should receive at least twelve (12) administrations of Foxy-5 monotherapy (1.8 up to 8 mg/kg, intravenously [IV], three to five administrations per week) prior to planned surgery.

Subjects randomized to the Control Arm will continue to receive standard of care only.

Safety will be observed and evaluated throughout the treatment period and for an additional 28 days after the last dose of Foxy-5.

Safety of subjects is monitored throughout the trial by a Data Safety Monitoring Board (DSMB).

RESULTS of Example 1

An ad-hoc analysis of 116 patients from part 1 of the NeoFox study (57 patients in the control arm and 59 patients in the Foxy-5 arm) was performed to understand possible trends. TNM at randomization as judged by CT/MRI was compared to TNM staging at surgery, based on the pathology reports (Table 1):

Of the 116 patients, 50 showed a downstaging in TNM between randomization and surgery, and 44 showed an upstaging. The remaining 22 patients either had no change in staging between the two time points or were not possible to evaluate (Figure 1 and 2).

With focus on the eligible 94 patients, the statistical analysis showed a significant association between staging and treatment based on a variety of tests. Data were also tested using a model based version of logistic regression, which showed that downstaging vs upstaging differed significantly between arms.

The conclusion is that downstaging was more frequent in the Foxy-5 arm. This was confirmed by significant association tests (p < 0.01) between change in staging and treatment arm. This was further confirmed by a logistic regression model-based method. Example 2

The in vivo effect of Foxy-5 on p-catenin signalling in HT-29 or Caco-2 colon cancer tissue could be an explanation for the observed downstaging discussed hereinabove. Decreased amounts of active p-catenin nuclei expression were observed in both Caco-2 and HT29 derived colon cancer tumours (see Figure 3 and 4) as a result of Foxy-5 treatment. The observations of Foxy-5-reduced p-catenin signalling were validated by reductions in tumour volumes (Figure 3 and 4).