Compounds, compositions and methods for treating, reversing or preventing cancer

Abstract

This invention relates to compounds, compositions and methods for treating, reversing or preventing cancer through the modulation of immune responses via Killer-specific Secretory Protein 37 (Ksp37). By enhancing immune recognition, Ksp37 downregulates the checkpoint proteins CD47 and CD24 on abnormal cells, allowing the immune system to effectively identify and eliminate cancerous cells. Additionally, Ksp37 influences cytokine signalling by increasing IL-12 and decreasing IL-2 levels, which supports the activation of natural killer (NK) cells while mitigating chronic inflammation, fostering an immune environment conducive to clearing abnormal cells.

Description

[0001] COMPOUNDS, COMPOSITIONS AND METHODS FOR TREATING,

[0002] REVERSING OR PREVENTING CANCER

[0003] FIELD OF THE INVENTION

[0004] This invention relates to compounds, compositions and methods for treating, reversing or preventing cancer and more particularly, but not exclusively, to compounds, compositions and methods for treating, reversing or preventing cancer through the modulation of immune responses via Killer-specific Secretory Protein 37 (Ksp37).

[0005] BACKGROUND TO THE INVENTION

[0006] Neoplastic disease, commonly referred to as cancer, involves the abnormal and uncontrolled proliferation of cells. This pathological growth may present as localised solid masses or may affect circulating cells and tissues, such as blood or lymph, without forming discrete masses. Cancer may arise in virtually any tissue type and is classified according to its cellular origin and biological behaviour. In healthy individuals, immune surveillance mechanisms play a critical role in detecting and eliminating aged, damaged, or aberrantly proliferating cells. Natural killer (NK) cells, as key effectors of the innate immune system, are particularly important in this process. NK cell-mediated clearance relies on a complex network of immune-regulatory signals that distinguish healthy "self1cells from those that are compromised. Two notable regulators of this process are the Cluster of Differentiation 47 (CD47) and Cluster of Differentiation 24 (CD24) receptors. These surface proteins form part of a “self-recognition” mechanism, conveying inhibitory signals to immune cells to prevent unnecessary immune-mediated destruction of healthy tissue.

[0007] Under physiological conditions, cells that become senescent, stressed, or otherwise abnormal downregulate these protective markers, permitting immune-mediated clearance. However, cancer cells frequently exploit this system of immune tolerance to escape detection and destruction. By overexpressing CD47 and CD24, malignant cells mimic healthy cells and suppress immune responses. CD47 binds to signal-regulatory protein alpha (SIRPa) on myeloid cells to inhibit phagocytosis, while CD24 engages Siglec- 10 on NK cells and macrophages, further dampening immune activity. This overexpression enables malignant cells to evade both innate and adaptive immune responses, facilitating unchecked proliferation and dissemination.

[0008] Beyond cell-surface checkpoint signalling, immunosuppressive cell populations within the tumour microenvironment also contribute to cancer progression. Regulatory T cells (Tregs), characterised by expression of CD25 and Forkhead Box P3 (FoxP3), are critical for immune homeostasis and the prevention of autoimmunity. However, their enrichment in the cancer setting has been associated with suppression of anti-cancer immunity and correlates with adverse clinical outcomes. Modulating Treg activity may therefore represent a valuable adjunct to strategies aimed at reversing immune evasion.

[0009] The ability of malignant cells to subvert immune regulation presents a significant challenge for cancer therapy. While conventional treatments — such as chemotherapy, radiotherapy, and surgical resection — may reduce cancer burden, they often lack specificity, result in substantial side effects, and fail to address the underlying immune escape mechanisms. A growing body of evidence highlights the therapeutic potential of targeting immune checkpoints and restoring immune surveillance.

[0010] Specifically, inhibition of the CD47 / SIRPa and CD24 / Siglec-10 signalling axes has emerged as a promising strategy for reactivating the immune system’s ability to eliminate malignant cells. Therapeutic interventions that interfere with these pathways may enhance NK cell cytotoxicity and macrophage-mediated clearance, thereby improving treatment outcomes.

[0011] Accordingly, there remains a pressing need for innovative therapeutic approaches that target the molecular mechanisms of immune evasion in cancer and restore effective immune surveillance. OBJECT OF THE INVENTION

[0012] It is an object of this invention to provide compounds, compositions and methods for treating, reversing or preventing cancer which, at least partially, alleviates some of the abovementioned difficulties.

[0013] SUMMARY OF THE INVENTION

[0014] The invention provides compounds, compositions, and methods for treating, reversing, or preventing cancer by raising the blood plasma concentration of Killer-specific Secretory Protein 37 (Ksp37) in a subject to a therapeutic level.

[0015] The compounds fall into two principal classes:

[0016] (i) a purified recombinant Ksp37 protein; and

[0017] (ii) polar compounds of general formula RI-CO-NR2R3, which promote endogenous expression of Ksp37.

[0018] The purified recombinant Ksp37 protein is expressed using a heterologous expression system, such as HEK293 or bacterial cells transfected with a plasmid encoding the native human Ksp37 gene. The plasmid may be based on the pcDNA3.1 backbone with the gene positioned downstream of a cytomegalovirus (CMV) promoter and upstream of a bovine growth hormone (BGH) polyadenylation signal. The protein is then harvested and purified to greater than 95% homogeneity for formulation into a pharmaceutical composition. The purified Ksp37 protein is not genetically modified or engineered and reflects the wild-type sequence.

[0019] Therapeutic efficacy is associated with raising Ksp37 plasma concentrations to between 300 ng / mL and 800 ng / mL, preferably around 600 ng / mL. This threshold is necessary to modulate immune function by altering cytokine signalling, enhancing immune cell-mediated cytotoxicity, and reducing cancer cell immune evasion.

[0020] The suitable polar compounds for stimulating endogenous Ksp37 expression include compounds of the general formula:

[0021] RI-CO-NR2R3, wherein:

[0022] Ri is selected from H, C1-C3 alkyl, or C2-C3alkenyl; and

[0023] R2and R3are independently selected from H, C1-C3 alkyl, or C2-C3alkenyl groups, optionally halogenated or hydroxylated, or forming a -CHn ring or -(CH2)2-O(CH)2group and their metabolites. Examples include, without limitation, N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), and N-ethylmaleimide (NEM). These compounds may be used independently or in sequence to achieve the desired therapeutic outcome.

[0024] These compounds may be administered independently or in sequence to achieve therapeutic modulation of the immune system through one or more mechanisms of action central to the invention.

[0025] Ksp37, whether delivered directly or produced endogenously following stimulation, functions in a subject as an immune effector molecule that restores effective immune surveillance and counteracts immune evasion strategies commonly employed by abnormal or uncontrolled cells, such as cancer cells.

[0026] The protein enhances the cytotoxic activity of natural killer (NK) cells to recognise and eliminate such cells.

[0027] In particular, Ksp37 facilitates immune recognition by suppressing the expression of Cluster of Differentiation 47 (CD47) and Cluster of Differentiation 24 (CD24) on abnormal cells, checkpoint proteins frequently overexpressed to escape immune detection. By downregulating these immune-evasion markers, Ksp37 inhibits immune checkpoint signalling pathways that normally prevent immune recognition and cytotoxic clearance. In addition to restoring visibility of abnormal cells to the immune system, Ksp37 modulates cytokine signalling, including upregulation of interleukin-12 (IL-12) and downregulation of interleukin-2 (IL-2). This cytokine shift supports activation and persistence of NK cells and other immune effector cells while simultaneously reducing excessive proliferation of effector T cells and curbing chronic inflammation. These changes contribute to an immune environment that is functionally oriented toward the clearance of abnormal or uncontrolled cells.

[0028] The invention also encompasses specific compounds and methods as claimed, including recombinant Ksp37 protein compositions, polar compound compositions, gene therapy vectors, acceptable excipients, and methods of administration via systemic, mucosal, or topical routes. The therapeutic plasma concentration of Ksp37 ranges between 300 ng / mL and 800 ng / mL, with a preferred level of 600 ng / mL, which is necessary for effective modulation of immune function, including cytokine signalling and immune cell-mediated cytotoxicity.

[0029] These and other features of the invention are described in more detail below.

[0030] DETAILED DESCRIPTION OF THE INVENTION

[0031] The invention provides compounds, compositions, and methods for treating, reversing, or preventing cancer by raising the blood plasma concentration of Ksp37 in a subject to a therapeutic level.

[0032] The compounds fall into two principal classes:

[0033] (i) a purified recombinant Ksp37 protein; and

[0034] (ii) polar compounds of general formula RI-CO-NR2R3, which promote endogenous expression of Ksp37.

[0035] The purified recombinant Ksp37 protein is expressed using a heterologous expression system, such as HEK293 or bacterial cells transfected with a plasmid encoding the native human Ksp37 gene. The plasmid may be based on the pcDNA3.1 backbone with the gene positioned downstream of a CM promoter and upstream of a BGH polyadenylation signal. The protein is then harvested and purified to greater than 90% homogeneity for formulation into a pharmaceutical composition. The purified Ksp37 protein is not genetically modified or engineered and reflects the wild-type sequence.

[0036] Therapeutic efficacy is associated with raising Ksp37 plasma concentrations to between 300 ng / mL and 800 ng / mL, preferably around 600 ng / mL. This threshold is necessary to restore effective immune surveillance and eliminate immune evasion mechanisms commonly used by cancer cells. The purified recombinant Ksp37 protein may also be produced using nonmammalian cell systems, such as fish skin cells transfected with a recombinant plasmid encoding the Ksp37 gene.

[0037] The suitable polar compounds for stimulating endogenous Ksp37 expression include compounds of the general formula:

[0038] RI-CO-NR2R3, wherein:

[0039] Ri is selected from H, C1-C3 alkyl, or C2-C3alkenyl; and

[0040] R2and R3are independently selected from H, C1-C3 alkyl, or C2-C3alkenyl groups, optionally halogenated or hydroxylated, or forming a -CHn ring or -(CH2)2-O(CH)2group and their metabolites.

[0041] Examples include, without limitation, N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), and N-ethylmaleimide (NEM). These compounds may be used independently or in sequence to achieve the desired therapeutic outcome.

[0042] These compounds may be administered independently or in sequence to achieve therapeutic modulation of the immune system through one or more mechanisms of action central to the invention. DMF, for example, may be administered transdermally via a patch system delivering approximately 9.6 grams of DMF over an 8-hour period, achieving an absorption rate of about 1.2 g / hour, and providing a dose of approximately 96 mg / kg for a subject weighing 100 kilograms.

[0043] Ksp37, whether delivered directly or produced endogenously following stimulation, functions in a subject as an immune effector molecule that restores effective immune surveillance and counteracts immune evasion strategies commonly employed by abnormal or uncontrolled cells, such as cancer cells.

[0044] The protein enhances the cytotoxic activity of natural killer (NK) cells to recognise and eliminate such cells. In particular, Ksp37 facilitates immune recognition by suppressing the expression of CD47 and / or CD24 on abnormal cells, checkpoint proteins frequently overexpressed to escape immune detection. By downregulating these immune-evasion markers, Ksp37 inhibits immune checkpoint signalling pathways that normally prevent immune recognition and cytotoxic clearance.

[0045] In addition to restoring visibility of abnormal cells to the immune system, Ksp37 modulates cytokine signalling, including upregulation of interleukin-12 (IL-12) and downregulation of interleukin-2 (IL-2). This cytokine shift supports activation and persistence of NK cells and other immune effector cells while simultaneously reducing excessive proliferation of effector T cells and curbing chronic inflammation. These changes contribute to an immune environment that is functionally oriented toward the clearance of abnormal or uncontrolled cells.

[0046] The invention is particularly applicable to a wide range of cancers, including but not limited to solid tumours such as breast cancer, colorectal cancer, glioblastoma, lung cancer, ovarian cancer, pancreatic cancer, and sarcomas. It also applies to blood and immune system cancers such as leukaemia, lymphomas, and myelomas, as well as cancers with elevated expression of immune checkpoint molecules or mechanisms of immune evasion.

[0047] The invention may be employed as a primary therapeutic approach, especially in cases where immune modulation is central to disease management. Additionally, it can be implemented as an adjunct therapy alongside existing cancer treatments. In this context, the Ksp37-based therapy may be combined with conventional approaches such as chemotherapy, radiotherapy, targeted biological treatments, or immune checkpoint inhibitors. Such combination strategies are designed to enhance the effectiveness of treatment regimens by improving immune surveillance and reducing tumour resistance.

[0048] By integrating Ksp37 therapy with established oncological practices, the invention addresses the limitations of monotherapies and enhances therapeutic outcomes, particularly in resistant or recurrent cancers. The flexibility of using Ksp37 either as a standalone intervention or in combination makes it a versatile tool in the oncologist’s arsenal.

[0049] The invention also encompasses specific compounds and methods as claimed, including recombinant Ksp37 protein compositions, polar compound compositions, gene therapy vectors, acceptable excipients, and methods of administration via systemic, mucosal, or topical routes. The therapeutic plasma concentration of Ksp37 ranges between 300 ng / mL and 800 ng / mL, with a preferred level of 600 ng / mL, which is necessary for effective modulation of immune function, including cytokine signalling and immune cell-mediated cytotoxicity.

[0050] It is envisaged that the invention described herein would be convenient to use as its ability to act at the intersection of innate and adaptive immune regulation makes it especially attractive for treatment across a broad spectrum of tumour types and clinical scenarios. Unlike conventional therapies that focus solely on tumour cytotoxicity, the invention re-establishes a balanced immunological environment that supports both immediate tumour clearance and long-term immune vigilance. The therapeutic modulation of cytokine signalling and checkpoint pathways provides a holistic means to overcome tumour immune escape. Moreover, the invention offers significant promise in the context of personalised medicine. Its flexible formulation options, capacity for real-time biomarker monitoring, and integration with both standalone and combination therapy protocols equip clinicians with a toolset to tailor treatment to individual patients. This precision approach reduces overtreatment, minimises systemic toxicity, and improves patient outcomes by enabling long-term disease control through immune restoration rather than immune suppression.

[0051] Ultimately, the invention offers a novel therapeutic strategy that is not only grounded in immunological science but also practically designed for real- world clinical use, addressing a pressing need for more effective and sustainable cancer therapies.

[0052] It will be appreciated by those skilled in the art that numerous variations and alternative embodiments are possible without departing from the scope of the present invention.

[0053] EXPERIMENTAL RESULTS

[0054] Experiment 1 : Effect of Dimethylformamide (DMF) on Tumour Cell

[0055] Phenotypes and Ksp37 Upregulation Background:

[0056] Studies investigating the effects of polar compounds on tumour cells have shown promising results in modulating cell behaviour and phenotypes. Specifically, dimethylformamide (DMF) has been observed to induce properties characteristic of benign, well-differentiated phenotypes in various tumour cell cultures. These changes, which are observable through alterations in physical properties, suggest a potential role for DMF in modifying tumour progression. DMF has been shown to cause conformational changes and hypomethylation of DNA sequences while inhibiting the expression of myconco-proteins. These proteins, encoded by oncogenes, are critical for the regulation and synthesis of proteins associated with cancer cell growth.

[0057] Materials and Methods:

[0058] To assess the effects of DMF, a variety of tumour cell lines were studied, including virus-transformed mouse kidney cells and human colon carcinoma cells. Cells were cultured in standard growth media, and experimental groups were supplemented with a 0.5% concentration of DMF. Observations were made on changes in growth patterns, cell morphology, and protein expression. DNA methylation levels were assessed using established molecular biology techniques, while protein expression was evaluated to identify changes in oncogene activity. Results:

[0059] Exposure to DMF resulted in significant changes in the growth patterns of tumour cells. Virus-transformed mouse kidney cells, which typically exhibited random piling and non-contact-inhibited growth, showed a marked shift in behaviour. When cultured with 0.5% DMF, these cells formed monolayers with a regular parallel orientation, resembling non-malignant fibroblasts. Similar phenotypic changes were observed in human colon carcinoma cells, where DMF exposure also led to more organised and less aggressive growth patterns.

[0060] Molecular analysis revealed that DMF induced hypomethylation of DNA sequences and inhibited the expression of myconco-proteins. These findings suggest that DMF impacts the epigenetic and proteomic landscapes of tumour cells, contributing to the observed changes in phenotype. Further research indicated that DMF’s effects were not direct but mediated through the upregulation Ksp37.

[0061] Ksp37 levels were found to be six- to eight-fold higher in circulating cancer cells (CCC) during early carcinoma stages compared to more advanced stages. Importantly, elevated Ksp37 levels correlated positively with improved clinical outcomes. This correlation was particularly evident in ovarian cancer, where Ksp37 levels strongly aligned with histology, tumour stage, and patient prognosis. Effect of Ksp37 on Natural Killer (NK) Cell Activation and

[0062] Proliferation

[0063] Background:

[0064] Natural Killer (NK) cells play a critical role in the body's defense against tumour cells by directly targeting and eliminating them. NK cells utilize various mechanisms to achieve this, including the release of cytotoxic granules containing perforin and granzymes, and the production of cytokines that activate other immune cells. The expression of CD69+ on NK cells is not only an indicator of activation but also a functional triggering molecule required for NK cell-mediated cytotoxicity against tumour cells.

[0065] In Vitro Data:

[0066] Human peripheral blood mononuclear cells (PBMCs) were processed to isolate CD8+ T cells and NK cells. These cells were cultured in RPMI 1640 media with varying concentrations of Ksp37 for 24 hours. The Ksp37 concentrations used in the first test were 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, and 0.125 pg / mL. In a subsequent test, concentrations of 8 pg / mL, 4 pg / mL, 2 pg / mL, and 0 pg / mL (as a control) were applied (see Figure 6 and Figure 7).

[0067] Activation levels of CD8+ T cells and NK cells were assessed using flow cytometry. The results demonstrated a consistently higher number of NK cells in wells containing Ksp37 compared to the control wells without Ksp37. Additionally, wells with Ksp37 exhibited a significantly greater number of CD69+ markers compared to the control. Since CD69+ is not only a marker of activation but also a functional molecule necessary for NK cell-mediated tumour cell killing, these results indicate that Ksp37 induces both proliferation and activation of NK cells. This suggests that Ksp37 enhances the immune system's ability to eliminate tumour cells through NK cell activation.

[0068] Table 1 - Results in Vitro Data

[0069] In Vivo Data: The experimental protocol involved infecting twenty-seven (27) humanized mice intraperitoneally with the Yu2 strain of HIV-1. Treatment commenced on day 43 (D43) post-infection. The mice were divided into the following treatment groups: Intravenous (i.v.) vehicle (PBS) as a control.

[0070] Intramuscular (i.m.) injection of Ksp37 plasmid resuspension solution on D43.

[0071] Antiretroviral therapy via food pellets from D43 to D71.

[0072] Weekly intravenous administration of purified Ksp37 protein at doses of 1.9,

[0073] 0.75, 0.25, and 0.083 mg / kg, respectively, starting on D43.

[0074] Intramuscular injection of Ksp37 plasmid on D43.

[0075] Plasma HIV-1 viral load and immune cell phenotyping were monitored using flow cytometry at the following time points:

[0076] . Week 5 (W5) I Day 36 (D36)

[0077] . Week 7 (W7) I Day 50 (D50)

[0078] . Week 8 (W8) I Day 57 (D57)

[0079] . Week 9 (W9) I Day 64 (D64)

[0080] . Week 10 (W10) / Day 71

[0081] The results indicated a trend of increased NK cell (CD56+) populations in the

[0082] Ksp37-treated groups compared to the control groups. This trend was particularly noticeable between weeks 8 and 10 of treatment. Similarly, there was a notable increase in CD69+ cells in the Ksp37-treated mice, suggesting that Ksp37 contributes to the activation and expansion of NK cells in vivo.

[0083] Table 2 - Results in Vivo Data

[0084] Conclusion:

[0085] The in vitro and in vivo data collectively suggest that Ksp37 has a significant effect on enhancing NK cell activation and proliferation. This activation is evidenced by the increase in CD69+ markers, both in cultured human cells and in treated mice. The observed enhancement of NK cell activity by Ksp37 indicates its potential therapeutic application in boosting immune surveillance and targeting tumour cells.

Claims

CLAIMS1. A compound comprising a recombinant Ksp37 protein, for use in treating, reversing or preventing cancer in a subject, wherein the compound is formulated to raise a blood plasma concentration of Ksp37 in the subject to between 300 ng / mL and 800 ng / mL.

2. The compound as claimed in claim 1 , wherein the recombinant Ksp37 protein is a purified recombinant Ksp37 protein.

3. The compound as claimed in claim 2, wherein the purified recombinant Ksp37 is produced using a heterologous expression system and is harvested and purified to greater than 90% homogeneity.

4. The compound as claimed in claim 3, wherein the heterologous expression system includes mammalian cells, including HEK293 cells, transfected with a pcDNA3.1 plasmid encoding the Ksp37 gene, positioned downstream of a cytomegalovirus CMV (CMV) promoter and upstream of a bovine growth hormone BGH (BGH) polyadenylation signal.

5. The compound as claimed in claim 3 to 4, wherein the heterologous expression system includes non-mammalian cell systems, including fishskin cells and algae cells, transfected with a recombinant plasmid encoding the Ksp37 gene.

6. A compound of general formula RI-CO-NR2R3, wherein Ri is selected from any one or more of the group consisting of hydrogen, Ci-C3alkyl, and C2-C3alkenyl; and R2and R3are independently selected from the group consisting of hydrogen, the Ci-C3alkyl, halogenated or hydroxylated, or forming a -CHnring or a -(CH2)2-O(CH2) group ,the C2- C3alkenyl, halogenated or hydroxylated or forming a -CHnring or a - (CH2)2-O(CH2) group, for use in treating, reversing or preventing cancer in a subject, wherein the compound raises a blood plasma concentration of Ksp37 in the subject to between 300 ng / mL and 800 ng / mL.

7. The compound as claimed in claim 6, wherein the compound is selected from the group consisting of N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), and N-ethylmaleimide (NEM).

8. The compound as claimed in any one of the preceding claims, wherein the blood plasma concentration of Ksp37 in the subject is 600 ng / mL.

9. The compound as claimed in any one of the preceding claims, wherein Ksp37 when at a blood plasma concentration level between 300 ng / mL and 800 ng / mL modulates immune function by altering cytokine signalling,enhancing immune cell-mediated cytotoxicity, and reducing cancer cell immune evasion in the subject.

10. The compound as claimed in claim 9, wherein the alteration of cytokine signalling includes increasing interleukin-12 and a decreasing interleukin- 2.

11. The compound as claimed in claims 9 to 10, wherein the enhancement of immune cell-mediated cytotoxicity includes activation or enhancement of natural killer cell activity.

12. The compound as claimed in claims 9 to 11 , wherein the reduction of cancer cell immune evasion includes downregulation or suppression of Cluster of Differentiation 47 and Cluster of Differentiation 24 expression on cancer cells.

13. The compound as claimed in claims 9 to 12, wherein the modulation of the immune response further includes promotion of regulatory T cell formation and restoration of immune homeostasis.

14. A composition for use in treating, reversing, or preventing cancer comprising an acceptable excipient, together with one or more compounds selected from the group consisting of a recombinant Ksp37 protein; a polar compound of the general formula RI-CO-NR2R3asdefined in any one of claims 1 or 5; or a gene therapy vector encoding Ksp37, wherein the composition is formulated to raise the blood plasma concentration of Ksp37 in a subject to a level between 300 ng / mL and 800 ng / mL.

15. The composition as claimed in claim 14, wherein the blood plasma concentration of Ksp37 in the subject is 600 ng / mL.

16. The composition according to any one of claims 14 to 15, wherein the gene therapy vector is any one or more of the group consisting of an adeno-associated virus, Adeno-Associated Virus serotype 8, a lentiviral vector, or a plasmid vector specifically designed for gene therapy, comprising a CMV promoter and BGH polyadenylation signal, wherein the Ksp37 gene is inserted between the CMV promoter and BGH polyadenylation signal within the multiple cloning site of a pcDNA3.1 plasmid backbone.

17. The composition according to any one of claims 14 to 16, wherein the acceptable excipient includes at least one carrier, diluent, stabiliser, solvent, or delivery agent.

18. A method for treating, reversing, or preventing cancer in a subject, the steps of raising the plasma concentration of Ksp37 in a subject to a level between 300 ng / mL and 800 ng / mL to modulate immune function byaltering cytokine signalling, enhancing immune cell-mediated cytotoxicity, and reducing cancer cell immune evasion in the subject.

19. The method as claimed in claim 18, wherein the blood plasma concentration of Ksp37 in the subject is 600 ng / mL.

20. The method as claimed in any one of claims 18 to 19, including the step of administering the compound or composition as claimed in any one of claims 1 , 6, or 14, either alone or in combination.

21. The method as claimed in claim 20 in which the compound or composition is administered via a systemic, mucosal, or topical route as a single dose or over a period of time.

22. The method according to any one of claims 20 to 21 , wherein the composition comprising DMF is administered to the subject transdermally via a single patch, wherein the patch delivers 9.6 grams of DMF over a period of 8 hours, corresponding to an absorption rate of 1.2 g / hour, and wherein the dose is 96 mg / kg for a subject weighing 100 kilograms.

23. The method as claimed in claim 22, wherein the DMF dosage is selected to achieve a Ksp37 plasma concentration level between 50 mg / L and 500 mg / L.

24. The method as claimed in claim 23, wherein the DMF dosage is selected to achieve a Ksp37 plasma concentration level of 120 mg / L.

25. The method as claimed in any one of claim 20 to 24, including the step of monitoring the level of blood plasma concentration of Ksp37 in the subject to maintain a blood plasma concentration of Ksp37 in the subject between 300 ng / mL and 800 ng / mL.

26. The method as claimed in any one of claims 18 to 25, wherein the compound or composition is administered in conjunction with other cancer treatment therapies, including chemotherapy, immune checkpoint inhibitors, or radiation therapy.

27. The compound, composition, and method as claimed in any of the preceding claims in which cancer includes breast cancer, colorectal cancer, glioblastoma, lung cancer, ovarian cancer, pancreatic cancer, sarcomas, leukaemia, lymphomas, myelomas and cancers with elevated expression of immune checkpoint molecules or mechanisms of immune evasion.

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