Compounds for use in macrophage reprogramming

Abstract

The present invention relates to compounds for use for treating and / or preventing tumors by virtue of the ability to reprogram tumor-associated macrophages M2 (TAM-M2) to tumor-associated macrophages M1 (TAM-M1).

Description

[0001] "Compounds for use in macrophage reprogramming"

[0002] DESCRIPTION

[0003] The present invention finds use in the field of medicine, and in particular in preventing and treating tumors and tumor metastases .

[0004] Inflammatory cells represent a key component in the regulation of the biological processes which characterize, in the body, the niche in which the tumor develops.

[0005] The inflammatory microenvironment is now recognized as a fundamental factor in the process of carcinogenesis.

[0006] The study of the microenvironment in which the tumor develops has thus become an aspect of extreme interest due to the presence in this microenvironment of pro- and anti-cancer mechanisms which can coexist, become unbalanced and / or become pro-cancerous.

[0007] The suppression of effective immunity in anti-tumor mechanisms and the modulation of inflammatory phenomena which promote tumor are now considered essential factors in the development and progression of cancer.

[0008] The formation of an inflammatory microenvironment in tumors can be the result of aberrations involving onco-genes or oncosuppressor genes.

[0009] Alongside these events, the anomalies which are created in the regulation and homeostasis of the immune system at the level of the tumor microenvironment are extremely relevant.

[0010] These anomalies are responsible not only for the f ormation / progression of the primary tumor, but also for the metastasis process, which is the leading cause of death from disease in patients with cancer.

[0011] In this scenario, macrophages represent an important component of the immune system.

[0012] They form the infiltrate in tumor masses, but also play a decisive role in the newly metastasizing niches.

[0013] Tumor-associated macrophages (TAMs) play a critical role in tumor formation / progression through the production of mediators, such as cytokines and / or growth factors.

[0014] During the last decade, it has been shown that TAMs can polarize into two different classes referred to as Ml and M2.

[0015] The M1 / M2 polarization model has been characterized and studied in many solid tumor study models.

[0016] In general, the TAM-M1 population is characterized by the expression of high levels of IL12 and IL23, the ability to produce significant amounts of ROS / RNS (reactive oxygen and nitrogen species) , and inflammatory cytokines.

[0017] TAM-Mls also have an active role in causing the immune system response through This (hl-type T lymphocytes) , which act in an anti-cancer sense.

[0018] Already at an extremely early stage (in the tumor niche) , as the tumor evolves and progresses, the macrophages can polarize towards the class referred to as TAM-M2.

[0019] TAM-M2s acquire different and alternative features, phenotype and activation state with respect to TAM-Mls.

[0020] The main differences comprise, for example, the variation in the expression of different receptors, the different ability to present the antigen, the variation in metabolic functions such as arginine metabolism and the different production of cytokines.

[0021] In particular, TAM-M2s show pro-tumor, pro-angiogenic activity and strong immuno-inhibitory phenotype.

[0022] This class of macrophages is characterized by the ability to produce high amounts of IL10 (immuno-inhibitory) and specifically express surface receptors such as CD163 and in particular CD206, which is the main receptor capable of binding carbohydrate molecules characterized by particular structures rich in mannose.

[0023] Having demonstrated the role of TAM-M2s in solid tumors (primary or metastatic disease) , they represent a new therapeutic target in treating solid tumors.

[0024] Currently, the most advanced and innovative therapeutic approaches aim to suppress the TAM-M2 population.

[0025] This can occur by virtue of the inhibition of the CSF-1 / CSF-1R signaling pathway, which is essential for the survival of all macrophage populations (even those essential for the normal operation of the immune system) .

[0026] Recently, mAbs (monoclonal antibodies) have been used in clinical trials to block CSF-1R, obtaining promising results only in combination with some chemotherapy drugs; however, this approach also leads to the elimination of non-tumor macrophages, effectively interfering with the patients' innate immunity system and probably also altering the adaptive response.

[0027] This evidence clearly raises concerns about the safety of this treatment . In line with the approach described above, trials have begun using trabectedin, a natural product derived from marine organisms .

[0028] Trabectedin interferes with DNA transcription and repair in tumor cells but also causes the TAMs' death through still-unknown molecular mechanisms.

[0029] In any case, the previous observations on the safety of the treatment carried out for mAbs developed for CSF-1R are also valid in this case.

[0030] Alternatively, drugs capable of blocking the recruitment of monocytes into tumors and their polarization into macrophages are also being tested.

[0031] This seems possible, interrupting the CCL2-CCR2 signaling pathway. The use of specific mAbs for CCL2-CCR2 shows the reduction of tumor growth and the improvement of patient survival; however, the discontinuation of anti-CCL2 therapy causes the rapid repopulation of the tumors by monocytes and a consequent acceleration in the progression of tumors with the phenomenon of metastasis being more aggressive and exacerbated.

[0032] Currently, in order to achieve the same objective, clinical protocols are being studied which are based on blocking the signaling pathways of PD-1, PD-L1 which, however, are not only specific TAM markers but also of other cells of the immune system.

[0033] Non-specific macrophage population suppression can be risky, also considering the fact that these cells are key effectors in the mechanisms of antibody-mediated cellular cytotoxicity / phagocytosis, which represents a fundamental aspect in the mechanisms of the immune response in healthy subjects.

[0034] To overcome these problems, attempts are being made to develop new drugs which can specifically target TAM-M2s and have the ability to cause the death of these cells. However, the strategies adopted for this purpose have so far provided mixed results.

[0035] Very recently, a more attractive strategy for cancer therapy has been suggested, which is based on the functional reprogramming of TAMs .

[0036] In particular, it is the objective to generate new drugs capable of acting specifically on TAM-M2s, reprogramming the phenotype, metabolism, the ability to produce cytokines and cytotoxic molecules thereof, as TAM-Mls are instead capable of doing .

[0037] The stimulation of the immune system would thus be obtained, in particular modulating innate immunity and stimulating the cytotoxic cellular component, converting pro-tumor TAM-2s to antitumor TAM-ls without the risk of significantly altering the other components of the immune system, indeed causing them to recognize and fight tumor cells.

[0038] Summary of the invention

[0039] The inventors of the present patent application have surprisingly found compounds capable of specifically binding TAM- M2 macrophages and reprogramming the phenotype, metabolism, and ability thereof to produce cytokines, restoring the typical features of TAM-Mls ( anti-tumor ) ; such an ability surprisingly results in reducing the growth of the primary tumor and blocking the metastasis process of solid tumors.

[0040] Object of the invention

[0041] In a first object, a compound is described having the amino acid structure of SEQ. ID NO: 1 or 2 for the medical use.

[0042] In a particular aspect, such a medical use is described for preventing and / or treating and / or delaying the development of a tumor .

[0043] In another aspect, such a medical use is described for reprogramming the tumor-associated macrophages M2 (TAM-M2) to tumor-associated macrophages Ml (TAM-M1) .

[0044] In a further aspect, such a medical use is described for tumor-associated macrophages M2 (TAM-M2) present in the tumor niche .

[0045] A second object is represented by the process for preparing the compounds of the present invention.

[0046] According to a third object, a compound obtainable and a compound obtained with the process of the invention are described.

[0047] In accordance with a fourth object, a compound obtainable and a compound obtained with the process of the invention are described for the medical uses indicated above.

[0048] A compound having the amino acid structure of SEQ. ID NO: 1 or 2 is also described as such.

[0049] In a further object of the invention, a method for preventing and / or treating and / or delaying the development of tumor metastases is described.

[0050] Brief description of the drawings Figure 1 is a graph reporting the results of the MTT viability assay of the macrophages treated with MANI and MAN2.

[0051] Figure 2 shows the results of the differentiation assay of M2 macrophages in the Ml line for increased expression of the iNos marker .

[0052] Figure 3 shows the results of the differentiation assay of M2 macrophages in the Ml line for increased expression of the HLA-DR marker .

[0053] Figure 4 shows the results of the differentiation assay of M2 macrophages in the Ml line for decreased expression of the CD206 marker .

[0054] Figure 5 shows the results of the assay on IL10 production in macrophages previously differentiated into M2.

[0055] Figure 6 shows the results of the assay on IL12 production in macrophages previously differentiated into M2.

[0056] Figure 7 shows the results of the assay on the anti-metastatic effect of MANI in a mouse model of melanoma.

[0057] Figure 8 shows the results of the assay on the anti-metastatic effect of MANI in a triple negative mouse model of breast tumor. Detailed description of the invention

[0058] In a first object, a compound is described having the amino acid structure of SEQ. ID NO: 1 or 2 for the medical use.

[0059] In particular, in a first aspect, such a compound is represented by the compound comprising the structure of the sequence of SEQ. ID No. 2: In a preferred aspect, such a compound has a structure corresponding to the amino acid sequence of SEQ. ID No. 2.

[0060] In a second aspect, such a compound is represented by the compound comprising the structure of the sequence of SEQ. ID No. 1 :

[0061] In a preferred aspect, such a compound has a structure corresponding to the amino acid sequence of SEQ. ID No. 1.

[0062] In particular, a compound according to the invention is obtained from appropriately transformed plant or mammal cells.

[0063] The process for preparing the compound of the invention represents a second object of the present patent application.

[0064] The preparation of the compound of the invention can be obtained for production in both plant cells and mammal cells, as further detailed in the following Experimental Section.

[0065] In particular, plant cells can be appropriately transformed by means of the sequence of SEQ. ID No. 3: In this respect, for example, Nicotiana benthamiana cells can be transformed.

[0066] For example, the process can comprise the use of Agrobacterium tumefaciens .

[0067] In an aspect of the invention, the cultivation of the transformed plant cells is carried out in suspension.

[0068] In particular, mammal cells can be appropriately transformed by means of the sequence of SEQ. ID No. 4 :

[0069] In this respect, CHO cells (Chinese hamster ovary cells} can for example be transformed.

[0070] The process can for example comprise the use of Lipof ectamine technology .

[0071] The compound obtainable or the compound obtained by means of the transformation process in plant cells or in mammal cells as described herein represent a third object of the present invention .

[0072] According to the present invention, the medical use of a compound having the amino acid structure of SEQ. ID NO: 1 or 2 (as above) is described, as well as the medical use of a compound obtainable or obtained according to the process described above.

[0073] According to a particular aspect of the invention, for both compounds such a medical use is described for preventing and / or treating and / or delaying the development of a tumor.

[0074] According to another aspect of the invention, said tumor form is represented, for example, by: breast tumor, ovarian tumor, melanoma, neuroblastoma (also in pediatric age) , pancreatic tumor, colon tumor, uterine tumor, liver tumor, prostate tumor, kidney tumor, lung tumor.

[0075] In particular, breast tumor includes triple negative breast tumor, i.e., the cells of which do not have estrogen (ER) or progesterone (PR) receptors and without an increase in HER2 expression .

[0076] For the purposes of the present invention, preventing the tumor comprises preventing the development of a not yet-diagnosed neoplasm.

[0077] For the purposes of the present invention, treating the tumor comprises reducing the tumor mass, even until the disappearance thereof .

[0078] For the purposes of the present invention, delaying tumor development comprises slowing tumor development.

[0079] According to a particular aspect, said medical use is for preventing and / or treating and / or delaying the development of tumor metastases. For the purposes of the present invention, preventing the tumor comprises preventing the development of a not yet-diagnosed neoplasm.

[0080] For the purposes of the present invention, treating a metastasis comprises reducing the metastatic tumor mass, even until the disappearance thereof.

[0081] For the purposes of the present invention, delaying the development of a metastasis comprises slowing the development of the metastatic tumor mass.

[0082] The compounds of the invention thus have anti-metastatic activity.

[0083] In another aspect, such a medical use is described for reprogramming tumor-associated macrophages M2 (TAM-M2) to tumor- associated macrophages Ml (TAM-M1) .

[0084] In a further aspect, such a medical use is described for tumor-associated macrophages M2 (TAM-M2) present in the tumor niche .

[0085] The term "tumor niche" refers to the microenvironment where the tumor forms and grows. It is characterized by the presence of particular cytokines produced by tumor cells and healthy cells which are caused to modify the phenotype thereof, producing a series of essential factors for tumor development. These factors affect the neo-vascularization process, depress the immune system and make the surrounding tissues easily infiltratable by tumor cells . For the purposes of the present invention, a compound having the amino acid structure of SEQ. ID No. 1 or 2 is also described as such.

[0086] In a further object of the invention, a method for preventing and / or treating and / or delaying the development of tumor metastases is described.

[0087] In particular, such a method comprises administering to a subject in need thereof a pharmaceutically effective amount of a compound of the present invention.

[0088] Experimental section

[0089] Macrophage viability assay

[0090] A macrophage viability assay was conducted by means of MTT assay.

[0091] As the results of the assay show, neither MANI nor MAN2 alter macrophage viability.

[0092] Culture and transformation of plant and animal cell lines

[0093] Nicotian benthamiana plant cells were grown in MS (Murashige and Skoog) culture medium and binary plasmids derived from pGreen II were used for the transformation thereof. The transformation occurred using the classic process which includes using Agrobacterium tumefaciens to make the exogenous DNA enter the plant cell.

[0094] The plant cells were transformed by inserting the sequence of SEQ. ID No. 3.

[0095] The mammal cells (CHO) are cultured in suspension in ProCHO-4 culture medium (added with 4% L-Gln, 2% HT Supplement and 0.5% antibiotics) and pcDNA 3.1 plasmids were used for the transformation thereof . For the transformation of the animal cells, the chemical approach based on Lipof ectamine was used following the manufacturer' s instructions .

[0096] The mammal cells were transformed by inserting the sequence of SEQ. ID No. 4 .

[0097] Purification of MANI from plant cells

[0098] The transformed plant cells are harvested and separated from the culture medium, then lysed in the buffer: 50 mM Na2HPCO4, 150 mM NaCl, 20 mM citric acid, 40 mM ascorbic acid, 1 mM ethylenediaminetetraacetic acid (EDTA) , 1 mM phenylmethanesulfonyl fluoride (PMSF) , 2 mM potassium metabisulfite, 0.1% (w / v) benzoic acid, 0.05% (v / v) Tween-20, pH 6.5 added with 1% (w / v) XAD-4 and 1% (w / v) polyvinylpolypyrrolidone (PVPP) for 1 h at 4°C under continuous stirring. The extract is then centrifuged at 18000 g for 20 min at 4 °C to recover the supernatant.

[0099] Ammonium sulfate is added to the extract until a saturation concentration of 70% is obtained. After centrifugation at 18000 g for 20 min at 4°C, the precipitate is resuspended in 400 ml of buffer (20 mN Na2HPO4, 200 mM NaCl, 10 mM imidazole, 0.1% (w / v) benzoic acid, pH 8.0) suitable for promoting the interaction between the His-tag-derivatized protein and the IMAC resin.

[0100] The MANI protein is then purified from the centrifuged and filtered solution (0.22 pm) through subsequent chromatographies. The product eluted from the first affinity chromatography (IMAC binding HisTag) on HiTrap Chelating HP column (following the manufacturer's instructions, GE Healthcare) is desalted using Sephadex G-25 Medium resin (GE Healthcare) equilibrated and eluted with a buffer of 50 mM sodium acetate, 150 mM NaCl, 60 pM Tween-

[0101] 20, pH 5.5. The eluate from the desalting column is loaded on a column with SP Sepharose HP resin (GE Healthcare) equilibrated with a buffer of 50 mM sodium acetate, 150 mM NaCl, 60 pM Tween-

[0102] 20, pH 5.5 and eluted with a buffer of 50 mM sodium acetate, 1 M

[0103] NaCl, 60 pM Tween-20, pH 5.5 by means of a step gradient so as to obtain purified MANI. Purification of MAN2 from mammal cells

[0104] The transformed cells are harvested by centrifugation, the culture medium is removed and then lysed at 4°C for 1 h in a buffer of 50 mM Na2HPO4, 1 M NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA) , 1 mM phenylmethanesulfonyl fluoride (PMSF) , 0.05% (v / v) Tween-20, pH 7.2 at 4 °C under continuous stirring. The extract is then centrifuged at 18000 g for 20 min at 4°C to recover the supernatant. The sample is loaded onto HiTrap Chelating HP column (GE Healthcare) previously loaded with 0.1 M NiC12 solution and equilibrated with a buffer of 50 mM potassium phosphate, pH 7.2, 1 M NaCl, 20 mW imidazole. The protein was eluted from the chromatographic column by means of step gradient of elution buffer (50 mM potassium phosphate, pH 7.2, imidazole) . The purified MAN2 protein is loaded onto Sephadex G-25 column equilibrated in PBS buffer, pH 7.4, so as to remove the imidazole.

[0105] Preparation of human monocytes

[0106] The monocytes needed for the trial were isolated from human peripheral blood. The fresh plasma (buffy coat) was centrifuged at 3000 x g for 15 minutes at room temperature. The monocytes were prepared with the batch gradient centrifugation technique. 35 ml of buffy coat are deposited on a 15 ml cushion of Ficoll solution (1.077 g / ml) and then centrifuged at 400 x g for 30 min at room temperature. The white ring of mononuclear cells is harvested and transferred to a new tube to which an excess of PBS-EDTA (1 mM) is added and centrifuged at 300 x g for 10 min at room temperature. The cell pellet is then resuspended in RPMI-1640 medium without phenol red, added with 10% complement-inactivated FCS.

[0107] The second density gradient consists of a Percoll cushion (23.13 ml of Percoll solution (density 1.131 g / ml) mixed with 1.87 ml of 10X PBS diluted 1:2 in RPMI-1640) . 25 ml of this solution are placed in a 50 ml tube, PBMCs (Peripheral Blood Mononuclear Cells) are deposited above such a solution and centrifuged at 550 x g for 30 min at room temperature. The white ring between the two phases of different density consists of monocytes, and is taken and resuspended in PBS-EDTA (1 mW) . Centrifuge at 400 x g for 10 min, the cell pellet is then resuspended in RPMI-1640 medium with phenol red added with 10% complement-inactivated FCS.

[0108] Differentiation of human monocytes into Ml or M2 macrophages

[0109] The isolated monocytes are seeded into culture dishes using medium consisting of RPMI-1640, 2% human serum, and 1% penicillin / streptomycin . The plates are kept in incubator (37°C with 5% CO) for 4-5 days before causing differentiation. Cell differentiation is caused by treating the cells for 10 days with 2 ng / ml GM-CSF (Ml differentiation) or 2 ng / ml M-CSF (M2 differentiation) .

[0110] Effect of MANI and MAN2 on macrophage physiology

[0111] The effect of MANI and MAN2 ligands on in vitro differentiated human macrophages was assessed through the analysis of the IL12 and IL10 production, which are produced in different amounts from the type Ml or M2 macrophages. The interleukins were measured in the culture media of the treated cells using specific ELISA kits and following the manufacturer's instructions.

[0112] The differentiated macrophages were analyzed using several antibodies specific for molecular markers characteristic of monocyte differentiation into:

[0113] - macrophages: CD68 (co-marking) ,

[0114] - type 1 macrophages: iNOS and HLADR,

[0115] - M2 macrophages: CD206. Flow cytof luorimetry (FACS) was used for this purpose. The cells (macrophages differentiated and treated with the different molecules) were harvested from the culture flasks and kept on ice in flow cytometry tubes throughout the experimentation period. The cells were incubated with several primary antibodies, each specific for a given receptor among those listed above. The detection is made possible by virtue of the use of secondary antibodies conjugated with different fluorophores so that multiplex analysis can be performed on the cytof luorimeter .

[0116] The results of the assays are shown in the figures.

[0117] In particular, figure 2 shows a significant increase in the expression of the iNOs marker, characteristic for Mis.

[0118] Figure 3 shows a significant increase in the expression of the

[0119] HLA-DR marker, characteristic for Mis.

[0120] Figure 4 shows a significant reduction in the expression of the

[0121] CD206 marker, characteristic for M2s.

[0122] Figure 5 shows the blockage of the production of the IL10 marker characteristic for M2s following inflammatory stimulus (LPS) typical of M2 macrophages in macrophages previously differentiated into M2s and treated with MANI or MAN2.

[0123] Figure 6 shows the results of incubation of the M2-dif f erentiated macrophages with MANI or MAN2 ligands, leading to increased expression of the IL12 marker, typical of Mis.

[0124] Effect of MANI or MAN2 ligand as anti-metastatic

[0125] Two syngeneic mouse models of particularly aggressive metastatic tumors were used to evaluate the in vivo anti-metastatic effect of the MANI ligand. a) Mouse model of melanoma

[0126] B16F10 cells (murine melanoma model) were used, inoculated into the animals in the suprascapular sub-skin. The cells generate a very aggressive primary tumor, which quickly generates many metastases which lead to the death of the host. The metastases originate very early in the peritoneum and lungs. The metastases can be easily detected, as the melanin-rich cells are easily identified. The experiment includes the comparison between two experimental groups: one control group (not treated with MANI) and the group treated with MANI. The ligand was administered to the animals intraperitoneally at a dosage of 2 mg / kg twice a week for the entire duration of the experiment (3 weeks) . The administration of MANI was initiated immediately after inoculation of the tumor cells into the animal. The presence of metastases was assessed through necroscopic examination.

[0127] The results are shown in figure 7. b) Triple negative mouse model of breast tumor

[0128] E0771 cells (mouse model of triple negative breast tumor) were used, inoculated in animals into the fat of the mammary gland (orthotopic model) . The MANI ligand was administered to the animals intraperitoneally at a dosage of 2 mg / kg twice weekly for the entire duration of the experiment (8 weeks) , until the time of animal sacrifice. The administration of MANI was initiated immediately after inoculation of the tumor cells into the animal. After the inoculation of the cells, the animals are monitored daily to verify the growth of the primary tumor. When the tumor reaches the appropriate size (10x10 mm) , it is surgically removed. The animals are then reared for 45 days after removal of the primary tumor and then euthanized. The search for metastases at the pulmonary level is carried out through necropsy analysis.

[0129] The results are shown in figure 8.

Claims

CLAIMS1. A compound having the amino acid sequence of SEQ. ID NO: 1 or 2 for the medical use.

2. A compound for the medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor.

3. A compound for the medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor, wherein said tumor is a primary tumor or a tumor metastasis.

4. A compound for the medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor, wherein said tumor is represented by: breast tumor, ovarian tumor, melanoma, neuroblastoma, pancreatic tumor, colon tumor, uterine tumor, liver tumor, prostate tumor, kidney tumor, lung tumor.

5. A compound having the amino acid sequence of SEQ. ID NO: 1 or 2 for the medical use according to claim 1, wherein the tumor- associated macrophages M2 (TAM-M2) are reprogrammed to tumor- associated macrophages Ml (TAM-M1) .

6. A compound having the amino acid sequence of SEQ. ID NO: 1 or 2 for the medical use according to the preceding claim, wherein said tumor-associated macrophages M2 (TAM-M2) are macrophages present in the tumor niche.

7. A process for preparing a compound having the amino acid sequence of SEQ. ID NO: 1 or 2, wherein plant cells aretransformed by inserting the sequence corresponding to SEQ. ID NO:3.

8. A process according to the preceding claim, wherein said plant cells are Nicotians benthamiana cells.

9. A process for preparing a compound having the amino acid sequence of SEQ. ID NO: 1 or 2, wherein mammalian cells are transformed by inserting the sequence corresponding to SEQ. ID NO:4.

10. A process according to the preceding claim, wherein said mammalian cells are CHO cells (Chinese hamster ovary cellsthe ) .

11. A compound having the amino acid sequence of SEQ. ID NO: 1 or 2.

12. Use of a compound having the amino acid sequence of SEQ. ID NO: 1 or 2 for reprogramming tumor-associated macrophages M2 (TAM-M2) to tumor-associated macrophages Ml (TAM-M1) .

13. A compound being obtainable by the process according to any one of the preceding claims 7 to 10.

14. A compound being obtainable by the process according to any one of the preceding claims 7 to 10 for the medical use.

15. A compound for the medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor.

16. A compound for medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor, wherein said tumor is a primary tumor or a tumor metastasis.

17. A compound for the medical use according to the preceding claim for preventing and / or treating and / or delaying the development of a tumor, wherein said tumor is represented by: breast tumor, ovarian tumor, melanoma, neuroblastoma, pancreatic tumor, colon tumor, uterine tumor, liver tumor, prostate tumor, kidney tumor, lung tumor.

18. A compound being obtainable by the process according to any one of the preceding claims 7 to 10 for the medical use, wherein the tumor-associated macrophages M2 (TAM-M2) are reprogrammed to tumor-associated macrophages Ml (TAM-M1) .

19. A compound being obtainable by the process according to any one of the preceding claims 7 to 10 for the medical use, wherein said tumor-associated macrophages M2 (TAM-M2) are macrophages present in the tumor niche.

20. A method for the prevention and / or treatment and / or delay of the development of tumors or tumor metastases comprising the administration to a subject in need thereof of a pharmaceutically effective amount of a compound according to claim 1.

21. A method for preventing and / or treating and / or delaying the development of tumors or tumor metastasis according to the preceding claim, wherein the tumor-associated M2 macrophages (TAM- M2 ) are reprogrammed to tumor-associated Ml macrophages tumor(TAM-M1) .

22. The method for preventing and / or treating and / or delaying the development of tumors or tumor metastases according to the previous claim, wherein said tumor-associated M2 macrophages (TAM- M2 ) are present in the tumor niche.

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