WO2012123774A1

WO2012123774A1 - Glycolytic inhibitor with cytotoxic agent for use in the treatment of a cancer

Info

Publication number: WO2012123774A1
Application number: PCT/IB2011/000795
Authority: WO
Inventors: Jean-Ehrland RICCI
Original assignee: INSERM (Institut National de la Santé et de la Recherche Médicale)
Priority date: 2011-03-11
Filing date: 2011-03-11
Publication date: 2012-09-20

Abstract

The present invention relates to a glycolytic inhibitor for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a cytotoxic agent is administered.

Description

GLYCOLYTIC INHIBITOR WITH CYTOTOXIC AGENT

FOR USE IN THE TREATMENT OF A CANCER

FIELD OF THE INVENTION

The present invention relates to a method for use in the treatment cancer.

BACKGROUND OF THE INVENTION

Chemotherapeutic cytotoxic agents are the main clinical tool to control invasive malignancy. The classes of cytotoxic agents used most widely in the oncology clinic are DNA-damaging agents; antimetabolites ; and antimitotics (Cancer immunotherapy : immune suppression and tumor growth, George C. et al.) (Chemotherapy at Dorland's Medical Dictionary)).

DNA-damaging agents :

DNA- damaging agents may be alkylating agents, topoisomerase inhibitors or platinum compounds.

-Alkylating agents (Anatomical Therapeutic Chemical Classification (ATC) code L01A) are so named because of their ability to alkylate many nucleophilic functional groups under conditions present in cells. They impair cell function by forming covalent bonds with the amino, carboxyl, sulfhydryl, and phosphate groups in biologically important molecules.

Examples of alkylating agents are cyclophosphamide, chlorambucil, chlormethine, busulfan, treosulfan and thiotepa.

-Topoisomerase inhibitors (ATC code LOICB and L01XX) are agents that blocks type I or type II topoisomerases interfering thus with both transcription and replication of DNA by upsetting proper DNA supercoiling.

Examples of type I topoisomerase inhibitors are camptothecins, irinotecan and topotecan. Examples of type II inhibitors are amsacrine, etoposide, etoposide phosphate, and teniposide.

-Platinum compounds damage DNA by creating intrastrand and interstrand cross-links.

Examples of platinum compounds are cisplatin, carboplatin, oxaliplatin.

Anti-tumor antibiotics :

Anti-tumor antibiotics include mainly anthracyclines.

-Anthracyclines (ATC code : L01DB )

Examples of anthracyclines are doxorubicin, epirubicin, idarubicin, mitoxantrone, valrubicin.

-Other anti-tumor antibiotics (ATC code : L01DC)

Examples of other anti-tumor antibiotics are mitomycin, bleomycin and plicamycin. Anti-metabolites (ATC code L01B) :

Anti-metabolites are similar in structure to naturally occurring compounds that are required for the viability and division of a cell. The efficacy of the most important anti-metabolites against a range of tumor cells is based on the inhibition of purine or pyrimidine nucleoside synthesis pathway that are required for DNA synthesis.

Anti-metabolites can be divided into several classes, including folate antagonists such as methotrexate, purine antagonists such as fludarabine and pyrimidine antagonists such as 5-fluorouracil.

Antimitotics :

-Taxanes (ATC code : L01CD) interfere with microtubules. They block cell growth by stopping mitosis.

Examples of taxanes are paclitaxel, and docetaxel. -Vinca alkaloids (ATC code : L01CA) bind to specific sites on tubulin, inhibiting the assembly of tubulin into microtubules (M phase of the cell cycle). They are derived from the Madagascar periwinkle, Catharanthus roseus (formerly known as Vinca rosea).

Examples of vinca alkaloids are vincristine, vinblastine, vinorelbine,vindesine.

Conventional anticancer chemotherapies are generally thought to reduce tumor progression by direct cytostatic and cytotoxic effects on tumor cells.

However, even after an initial therapeutic success, some patients fail to display a clinically relevant antitumor immune response and succumb to tumor cell variants that escape from chemotherapy. (Casares et al. (2005); Obeid et al. (2007))

To completely and permanently succeed against cancers, cancer therapies must target all tumor cells, including cancer stem cells or they must remove a fraction of the tumor and be accompanied by a "bystander effect" in which the immune system recognizes, attacks and eradicates the remaining tumor cells including cancer stem cells and chemotherapy resistant cancer cells.

There is a growing consensus that the use of an individual chemotherapeutic cytotoxic agent is unlikely to succeed as a therapeutic strategy in the long run. The best approach to circumvent such problem would be to identify a way not only to induce an efficient and specific death of the tumor cells but also to educate the immune system to react against any remaining tumor cells; thereby extending the cytotoxic effect of the chemotherapy and preventing metastasis apparition.

SUMMARY OF THE INVENTION The inventors have demonstrated that a treatment combining a glycolytic inhibitor and a cytotoxic agent will represent an ideal way to reach such goal, in particular in colorectal cancer, cervical cancer and melanoma The present invention relates to a method for the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject in need thereof comprising administering to said subject an effective amount of glycolytic inhibitor and an effective amount of cytotoxic agent.

The present invention also provides a kit for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma comprising : (a) glycolytic inhibitor ; and (b) a cytotoxic agent.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a method for the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject in need thereof comprising administering to said subject an effective amount of glycolytic inhibitor and an effective amount of cytotoxic agent.

In the method of treatment according to the invention, the presence of a glycolytic inhibitor, in combination with a cytotoxic agent, enables to extend the cytotoxic effect of the cytotoxic agent by inducing an anticancer immune response.

Glycolysis refers to the series of enzymatic reactions which convert a molecule of glucose into lactate with the generation of two molecules of ATP. Glycolytic inhibitors are being designed that target the enzymes involved in the glycolysis pathway.

Glycolytic inhibitors are well known in the state of art and some are developed in cancer therapy (see, for example, Pelicano H et al. (2006) ; Pathania D et al. (2009)) ; El Mjiyad N et al. (2011)).

Indeed, tumor cells undergo accelerated aerobic glycolysis (Warburg effect). This peculiar metabolism of the cancer cells has led the field to develop new therapeutic strategies using glycolytic inhibitors (Scatena et al., 2008). The present invention relates to a cytotoxic agent for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a glycolytic inhibitor is administered.

The present invention relates to the use of a glycolytic inhibitor for the preparation of a medicament for the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a cytotoxic agent is administered.

The present invention relates to the use of a cytotoxic agent for the preparation of a medicament for the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a glycolytic inhibitor is administered.

In one preferred embodiment, the subject is a subject with stage III or stage IV cancer.

In a particular embodiment, the cancer according to the invention is a colorectal cancer. In case of colorectal cancer, standard treatment with chemotherapy is quite effective in stage I disease and stage II disease. Indeed, 5 years survival rates approach 90% for colon cancer.

However, stage III disease denotes lymph node involvement, and studies indicate that the number of lymph nodes involved affects prognosis (55% to 20%: 5 year survival) and stage IV colon cancer (metastatic) clearly has the worst prognosis (<5% 5 year survival) (National Cancer Institute).

In one preferred embodiment, the subject is a subject with stage III or stage IV colorectal cancer.

In a particular embodiment, the cancer according to the invention is cervical cancer.

In case of cervical cancer, with treatment, the 5-year relative survival rate for the earliest stage of invasive cervical cancer is superior to 90%. This survival rate decreases importantly to 25 to 35% of women with stage III cancer and 15% or fewer of those with stage IV cancer are alive after 5 years.

In one preferred embodiment, the subject is a subject with stage III or stage IV cervical cancer.

In a particular embodiment, the cancer according to the invention is melanoma.

Melanoma confined to the skin is curable in 95% to 98% of cases but the prognosis become dismal when the lesion recurs, or spreads from the skin to the lymph nodes or distant organs. Indeed, until now, no treatment has increased survival significantly in melanoma patients with metastases. Average survival for Stage III and IV patients has been about eight months. In one preferred embodiment, the subject is a subject with stage III or stage IV melanoma.

In one embodiment, the cytotoxic agent according to the invention is a cytotoxic agent which does not induce an anticancer immune response.

A "cytotoxic agent which induces an anticancer immune response" refers to a cytotoxic agent which, upon administration to tumor cells induces the dying cells to express tumor antigen(s) which elicits a protective response when said dying cells are injected in an animal in the absence of any adjuvant (Casares et al., 2005; Martins et al., 2010). When living cells expressing the tumor antigen(s) is (are) injected into the animal, such a protective immune response precludes the growth of these living cells.

A "cytotoxic agent which does not induce an anticancer immune response" refers to the contrary to a cytotoxic agent which, when administered to cells, does not induce the dying cells to elicit a protective response in an animal in which they are injected (Casares et al., 2005; Martins et al., 2010).

One method for evaluating if an agent "induces an anticancer immune response" is well described by Casares N. et al. (2005).

Typically, cytotoxic agents which induce an anticancer immune response are anti- cancer agents which induce the expression of at least one tumor antigen selected from the group consisting of calreticuhn, Heat Shock Proteins (HSPs) or High-mobility group box 1 (HMGB1). The expression of said tumor antigen(s) can be detected by standard methods known in the art such as RT-PCR, immunoblot etc...

Typically, oxaliplatin and anthracyclines such as doxorubicin are known to be cytotoxic agents which induce an anticancer immune response (Obeid M. et al. (2007); Casares N. et al. (2005) ; Martins I. et al., (2010)). On the contrary, etoposide, cisplatin and mitomycin are known to be cytotoxic agents which do not induce an anticancer immune response (Obeid M. et al. (2007); Casares N. et al. (2005) ; Martins I. et al., (2010)).

In one embodiment, the glycolytic inhibitor according to the present invention is a hexokinase (HK) inhibitor.

Hexokinase catalyzes the first step of glycolysis, phosphorylating hexose (e.g. glucose) to hexose-6-phosphate.

Hexokinase inhibitors are well known in the state of art (see, for example,

Pelicano H et al. (2006) ; Pathania D et al. (2009)) ; El Mjiyad N et al. (2011)).

In a preferred embodiment, the glycolytic inhibitor is a HK inhibitor selected from the group consisting of 2-deoxyglucose, 5-thioglucose, mannoheptulose, 3- bromopyruvate, lonidamine and methyl jasmonate.

In another embodiment, glycolytic inhibitor is a glyceraldehyde-3 phosphate dehydrogenase (GAPDH) inhibitor.

GAPDH converts glyceraldehydes-3 -phosphate to 1,3-bisphospho-glycerate with a simultaneous reduction of NAD+ to NADH (nicotin-amide adenine dinucleotide; oxidized and reduced respectively).

In the state of art, there are many well known GAPDH inhibitors such as, for example, alpha-chlorhydrin, ornidazole, arsenic, iodoacetate, koningic acid (see, for example, Pelicano H et al. (2006) ; Pathania D et al. (2009) ; El Mjiyad N et al. (2011)). Several additional GAPDH inhibitors are currently under development. Preferably, glycolytic inhibitor is a GAPDH inhibitor selected from the group consisting of alpha-chlorhydrin, ornidazole, arsenic, iodoacetate, koningic acid.

In one embodiment, the cytotoxic agent is a topoisomerase inhibitor. In a preferred embodiment, the cytotoxic agent is a topoisomerase II inhibitor.

In a more preferred embodiment, the cytotoxic agent is a topoisomerase II inhibitor selected from the group consisting of amsacrine, etoposide, etoposide phosphate, and teniposide.

In another embodiment, the cytotoxic agent is a topoisomerase I inhibitor.

In another embodiment, the cytotoxic agent is an anti-tumor antibiotic.

In a preferred embodiment, the cytotoxic agent is an anti-tumor antibiotic selected in the group consisting from mitomycin, bleomycin and plicamycin.

In another embodiment, the cytotoxic agent is a platinum compound.

In a preferred embodiment, the cytotoxic agent is a platinum compound selected from the group consisting of cisplatin and carboplatin.

The present invention also relates to a composition for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma comprising a glycolytic inhibitor and a cytotoxic agent.

In one embodiment, the cytotoxic agent of the composition according to the invention is a cytotxic agent which doesn't induce anticancer immune response.

The present invention also relates to a kit for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma comprising :

a- a glycolytic inhibitor ; and

b- a cytotoxic agent.

In one embodiment, the cytotoxic agent of the kit according to the invention is a cytotxic agent which doesn't induce anticancer immune response. In one embodiment the glycolytic inhibitor and the cytotoxic agent are administered at the same time. In another embodiment, the glycolytic inhibitor and the cytotoxic agent are administered sequentially.

For example, they can be administrated at different moments of the day.

In the following, the invention will be illustrated by means of the following examples as well as the figures.

FIGURE LEGENDS

Figure 1 : Toxicity of various concentrations of Etoposide (ETO) with or without 2- deoxy-glucose (2DG) on Εμ-myc cells.

Figure 2 : Toxicity of various concentrations of Etoposide (ETO) with or without 2- deoxy-glucose (2DG) on CT26.

Figure 3 : Toxicity of various concentrations of Etoposide (ETO) with or without lonidamine (LND) on CT26.

Figure 4 : Toxicity of various concentrations of Mytomycin (mitom-c) with or without lonidamine (LND) on CT26.

Figure 5 : Toxicity of various concentrations of Mytomycin (mitom-c) with or without 3-bromopyruvic acid (3-BrPA) on CT26.

Figure 6 : Size of lymph nodes as a function of treatment with PBS, 2DG, ETO or ETO+2DG (IP 3times/week; 2DG : 500mg/kg; ETO 2.5mg/kg).

Figure 7 : Survival probability as a function of treatment with PBS, 2DG, ETO or ETO+2DG.

Figure 8 : Time of relapse as a function of treatment with ETO or ETO+2DG.

Figure 9 : Exposure of calreticulin as a function of treatment on different cell lines. EXAMPLE

Material and Methods:

Cell culture.

CT26, B 16 and HeLa cells were obtained from ATCC and cultured in RPMI 1640 or DMEM medium supplemented with 10% fetal calf serum, 1 mM pyruvate, 10 mM HEPES and 100 Units/ml each of penicillin and streptomycin. Cells were incubated at 37°C under 5% C02.

To induce cell death, cells were treated for 24 or 48 hours with the indicated amount of Etoposide (TRC), Mytomicin c (Sigma), Mitoxanthrone (Sigma), 2 deoxy glucose (TRC), 3-Bromopyruvic acid (Sigma) or Lonidamine (Tocris). Cell death measurements.

Cells were either trypsinized or centrifugated (Εμ-myc cells), wash in PBS and labelled with DAPI (Molecular Probes, 0.5 μg/ml). Cell death was analyzed immediately by flow cytometry and definied as DAPI+ cells using a MACSQuant Analyzer (Miltenyi Biotec).

Mice experiments

All mice were maintained in specific pathogen-free conditions and all experiments followed the guidelines of the federation of European Animal Science Association. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Centre Mediterraneen de Medecine Moleculaire (INSERM U895).

- Transgenic mice and transplantation of lymphomas

C57BL/6 Εμ-myc transgenic mice were purchased from Jackson Laboratories and were genotyped by PCR according to instructions from the supplier. Lymphoma bearing animals were killed by cervical dislocation as soon as any signs of illness can be detected. A single cell suspension was prepared from lymph nodes by teasing them on a 70 μπι nylon filter. Cells were either resuspended in complete medium (DMEM supplemented with 10% FCS, 10 mM HEPES, penicillin/streptomycin, 0.1 mM L-asparagine and 50 μΜ 2-mercaptoethanol) for further ex vivo analysis or directly reimplanted in wild type mice.

Lymphoma transfers were realized into syngenic, non-transgenic, 6-10 weeks old C57BL/6 females by tail vein injection (0.5xl0⁶ viable cells per mouse, in 200 μΐ sterile PBS). Recipient mice were monitored thrice a week for lymph nodes enlargement. As the inguinal lymph nodes reached 5 mm in the longest diameter, the animals were intraperitoneally injected, three times a week for the indicated period of time, with PBS, etoposide (2.5 mg/kg), 2DG (500 mg/kg) or both compounds and subsequently monitored for treatment response.

The survival of the mice and, for some experiments, the weight of the lymph nodes were measured. The "time of relapse" reflects the time between remission and re- palpability of a recurrent lymph-node enlargement.

- Antitumor vaccination

BALB/c height-week-old female mice (Harlan) were injected subcutaneously on the left flank with lxlO⁶ CT26 cells treated either with mitoxantrone, or with etoposide or with 2DG and etoposide until reaching 80% DAPI positive cells for each treatment. One week after, these mice were challenged by injecting 0.5x10⁶ live cells on the contralateral flank. The animals were then checked twice a week for tumor development using calipers.

Detection of ecto-calreticulin

For the assessment of calreticulin exposure, cells were first washed in cold PBS, fixed for 5 minutes at 4°C with 0.25% paraformaldehyde in PBS, washed again and stained with rabbit anti-mouse calreticulin antibody (1:200, Abeam), for 30 min at 4°C in 2% FCS PBS. After another wash, cells were incubated with anti-rabbit Alexa Fluor 488 conjugated secondary antibody (1:200, Molecular Probes) and the pellet was resuspended in PBS buffer with DAPI (0.5 μg/ml). Samples were then analyzed by MACSQuant Analyzer (Miltenyi Biotec) and checked for calreticulin staining on DAPI negative cells. CRT exposure to the plasma membrane was expressed in Mean of fluorescence intensity.

Results: - Glycolysis inhibition increases the sensitivity to cytotoxic agent-induced cell death.

In order to analyze the potentialization of cytotoxic agent-induced cell death, CT26 cell lines (murin colon carcinomas) and isolated primary non-Hodgkin lymphoma cells from Εμ-myc mice were used. Those transgenic animals are presenting a deregulated c-myc expression under the control of the immunoglobulin heavy chain (IgH) gene enhancer (Εμ) causes abnormal growth and proliferation of pre-B and B cells and prevents differentiation, culminating in clonal pre-B or immature B-cell lymphoma. Genetics and histopathology of Εμ-myc lymphomas resemble human non-Hodgkin's lymphomas (NHL) (Adams et al., 1985). Primary murin lymphoma or CT26 cells were treated with increasing amount of Etoposide in presence or absence of the glycolytic inhibitor 2-deoxyglucose (2DG) (Figure 1 and Figure 2). While glycolysis inhibition was leading to little toxicity on its own, in all cases, the cell death induction was synergized by the co-treatment, glycolytic inhibitor and cytotoxic agent, compared to each compound individually. Same results were obtained when CT26 cells were treated with Etoposide (ETO) and Lonidamine (LND) (Figure 3), Mitomycin C (Mitom-C) and Lonidamine (Figure 4) or Mitomycin C and 3- Bromopyruvic acid (3-BrPA) (Figure 5).

Subsequently the beneficial effect of the co-treatment was analyzed in vivo using the Εμ-myc model. For that matter WT syngenic immuno-competent C57BL6 mice received 0.5x10⁶ Εμ-myc cells I.V. 20 days later all mice presented enlarged lymph nodes indicating the induction of lymphoma. Then mice were treated with PBS, 2DG, Eto or Eto+2DG. In a first experiment, mice were sacrificed 21 days following the beginning of the treatment and lymph-nodes enlargement was measured. Co-treated animals were presenting significantly smaller lymph nodes than Eto- or control- treated mice. In order to analyze the impact of the co-treatment on the animal lifespan, the experiment was repeated and mice were treated until reaching a point were they had to be sacrificed for ethical concerns (end-point experiments). Figure 7 indicates that co-treated animals can leave 30% longer (69 vs 47 days) compared to the Eto group. Relapse of the disease is always observed in the Εμ-myc model upon treatment. It appeared that co-treated animal presented a time of relapse increase by 30% compared to Eto treated mice (Figure 8).

Therefore combining glycolytic inhibitor with standard cytotoxic agent is able to significantly protect mice.

- Cytotoxic agent combined with glycolytic inhibitor converts a non- immunogenic cancer cell death stimulus to an immunogenic one by controlling calreticulin exposure.

Altogether those results indicated that the co-treatment could increase the cytotoxicity induced by Etoposide or Mitomycin c. The inventors next wondered if the co- treatment could impact on the induction of an efficient anti-cancer immune response. For that matter, the inventors investigated CRT exposure on the surface of primary Εμ-myc cells. As suggested earlier (Obeid et al., 2007), Eto is a weak inducer of CRT exposure (Figure 9A). However, Eto+2DG is very efficient to do so. Mitoxantrone is used as a positive control for CRT exposure (Obeid et al., 2007). The inventors therefore extended this observation to 3 others cancer cell lines, Figure 9B: HeLa cells (Human cervical cancer cell line), 9C: B16 cells (murin melanoma cell lines) and 9D: CT26 (murin colon carcinoma cell lines). In each case, Eto+2DG was inducing a significant increase of CRT exposure compared to Eto or 2DG alone. Interestingly, an increase of CRT exposure to the plasma membrane could also be observed when Eto was combined with other hexokinase inhibitors: LND (Figure 9E) or 3-BrPA (Figure 9F).

To analyze the impact of the co-treatment on the induction of an efficient anti-cancer immune response, anti-tumoral vaccination essays were performed. For that matter, CT26 cells were killed by Eto, Eto+2DG or mitoxantrone (as a positive control) until reaching 80% dead cells in each condition. Then lxl0⁶dead CT26 cells were injected into one flank of BALB/c immuno-competent syngenic mice. One week later, 0.5x10⁶ of CT26 live cells were injected into the contra lateral flank of the mice. Therefore in absence of vaccination the mice will develop a tumor whereas if vaccinated, mice should be protected. Figure 10 described that, as expected, 100% of non-vaccinated mice were dying within 25 days following the challenge with alive CT26 cells. However, only 33% of the mice vaccinated with Eto-treated cells could survive up to 90 days. Very importantly, this percentage jump up to 66% in the group of mice vaccinated with Eto+2DG. Of note, the protection given by the co-treatment is very close to the one induced by mitoxantrone (75%).

These results show than combining glycolytic inhibitor with a classical cytotoxic agent is able to increase CRT exposure to the plasma membrane and therefore increase the anti-cancer immune response.

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Claims

1. A glycolytic inhibitor for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a cytotoxic agent is administered.

2. The glycolytic inhibitor according to claim 1 wherein cytotoxic agent is a cytotoxic agent which doesn't induce anticancer immune response.

3. The glycolytic inhibitor according to claim 1 or claim 2 wherein the glycolytic inhibitor is a hexokinase inhibitor.

4. The glycolytic inhibitor according to any of claims 1 to 3 wherein the glycolytic inhibitor is a hexokinase inhibitor selected from the group consisting of 2- deoxyglucose, 5-thioglucose, mannoheptulose, 3-bromopyruvate, lonidamine and methyl jasmonate.

5. The glycolytic inhibitor according to claim 1 or claim 2 wherein the glycolytic inhibitor is a glyceraldehyde-3 phosphate dehydrogenase (GAPDH) inhibitor.

6. The glycolytic inhibitor according to claim 1 or claim 2 wherein the glycolytic inhibitor is a GAPDH inhibitor selected from the group consisting of alpha- chlorhydrin, ornidazole, arsenic, iodoacetate, koningic acid.

7. The glycolytic inhibitor according to any of claims 1 to 6 wherein the cytototxic agent is a topoisomerase II inhibitor selected from the group consisting of amsacrine, etoposide, etoposide phosphate, and teniposide.

8. The glycolytic inhibitor according to any of claims 1 to 6 wherein cytototxic agent is an anti-tumor antibiotic selected in the group consisting from mitomycin, bleomycin and plicamycin.

9. The glycolytic inhibitor according to any of claims 1 to 6 wherein the cytotoxic agent is a platinum compound selected from the group consisting of cisplatin and carboplatin.

10. The glycolytic inhibitor according to any of claims 1 to 9 wherein the subject is a subject with stage III or stage IV cancer.

11. A cytotoxic agent for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma in a subject to whom a glycolytic inhibitor is administered.

12. A composition for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma comprising a glycolytic inhibitor and a cytotoxic agent.

13. The composition according to claim 12 wherein the cytotoxic agent is a cytotoxic agent which doesn't induce anticancer immune response.

14. A kit for use in the treatment of a cancer selected from the group consisting of colorectal cancer, cervical cancer and melanoma comprising :

a- a glycolytic inhibitor ; and

b- a cytotoxic agent.

15. The kit according to claim 14 wherein the cytotoxic agent is a cytotoxic agent which doesn't induce anticancer immune response.