Compounds for treating diseases

Abstract

The invention relates to an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

Description

[0001] COMPOUNDS FOR TREATING DISEASES

[0002] FIELD OF THE INVENTION

[0003] The invention relates to the field of treating diseases.

[0004] BACKGROUND OF THE INVENTION

[0005] Differentiated cluster 36 (CD36), a multi-ligand and multi-functional receptor, is also named as “SR-B2”. A variety of physiological processes are mediated by this membrane glycoprotein, which is found in hepatocytes, platelets, adipocytes, mononuclear phagocytes such as monocytes / macrophages, myocytes, brain microglia, astrocytes and some epithelial cells. CD36 recognizes many ligands including oxidized or modified low density lipoprotein (oxLDL, mLDL), long-chain fatty acids (LCFA), lipid and lipoprotein components of bacterial cell walls, thrombospondin (TSP) -1 and - 5, fibrillar 0-amyloid (fA0), and dying cells.

[0006] As a consequence of its broad expression and ability to recognize many ligands, CD36 participates in the regulation of many processes including inflammation, angiogenesis, native immunity, clearance of foreign and native substances from the body, and lipid metabolism. Increased CD36 expression / function is associated with several diseases and pathologies including atherosclerosis, stroke, and neurodegenerative diseases (Febbraio, M., et al., J Clin Invest 108:785-791(2001)). More recently, CD36 has been shown to hold dual activities as signaling receptor and fatty acid transporter regulating immune cell metabolism and fate (Chen, Y., et al., J Exp Med 219 (6):e20211314 (2022)).

[0007] Various compounds have been described that modulate CD36 functioning at the plasma membrane. The best-known examples are sulfo-N-succinimidyl derivatives of long-chain fatty acids, in particular palmitate (SSP) or oleate (SSO), which compounds have been instrumental for the discovery of the fatty acid-binding properties of CD36. However, the short half-life of SSP or SSO when in aqueous solution limits their use in vivo.

[0008] Therefore, there is a need to provide modulators of CD36 suitable for treating diseases.

[0009] BRIEF DESCRIPTION OF THE INVENTION In a first aspect, the invention relates to an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0010] In a second aspect, the invention relates to an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and C4BP isoforms lacking the beta chain wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury.

[0011] DESCRIPTION OF THE FIGURES

[0012] Figure 1.- Volcano plot showing the comparison between PRP6-HO7 (specific interaction(s)) and Glycine (nonspecific binding) samples. Adjusted p- values (plotted on logio scale) against the mean ratio protein fold-changes between samples (PRP6- HO7 / Gly) (plotted on log2 scale). The PRP6-HO7 receptor candidate space (upper right side) is defined by an enrichment factor of greater than twofold and an adjusted P value Figure 2.- PRP6-HO7 interaction with the surface of Mo-DCs. Saturation binding curve of fluorescence-labeled PRP6-HO7 to the surface of Mo-DCs assessed by flow cytometry. Data points are mean + SD of 4 separate donors. The binding was specific, cooperative, and of high-affinity (low nM range), suggesting a receptor clustering effect induced by PRP6-HO7.

[0013] Figure 3.- Multi-cycle kinetics showing surface plasmon resonance sensorgrams for PRP6-HO7 binding to CD36. Data are shown for 15 to 1,000 n PRP6-HO7 injected at 30 pl / min over the CD36 coated chip surface. Black lines: overlapping profiles according to the optimal fitting model tested. The x axis is time in seconds, the y axis is the surface plasmon resonance signal in response units (RU).

[0014] Figure 4.- PRP6-HO7 binds to CD36 in differentiating Mo-DCs. At 24 h of Mo-DC differentiation, cells were left untreated or treated with a CD36 blocking Ab or its isotype control, and further incubated by Strep-Tag Il-tagged PRP6-HO7. Specific PRP6-HO7-CD36 binding was visualized through a FITC-conjugated anti-Strep-Tag II and flow cytometry. The results shown are the mean ± SD from 5 independent donor samples (*, p < 0.05 compared with PRP6-HO7).

[0015] Figure 5.- Specificity of PRP6-HO7 and C4BP( -) binding to CD36. ELISA measurements of (A) immobilized hCD36-Fc or isotype control hlgGlK-Fc binding to PRP6-HO7, either alone or blocked (complexed) with soluble hCD36his. (B) immobilized hCD36-Fc, hCD36Ll-Fc, or isotype control hlgGlK-Fc binding to PRP6- HO7. (C) immobilized hCD36-Fc, mCD36-Fc, or isotype control hlgGlK-Fc binding to PRP6-HO7. (D) immobilized hCD36-Fc or isotype control hlgGlK-Fc binding to PRP6- HO7 or C4BP(0-), either alone or blocked (complexed) with soluble hCD36his. (E) immobilized hCD36-Fc, hCD36Ll-Fc, or isotype control hlgGlK-Fc binding to PRP6- HO7 or C4BP(0-). (F) immobilized hCD36-Fc, mCD36-Fc, or isotype control hlgGlK- Fc binding to PRP6-HO7 or C4BP(0-). Specific binding was assessed with an anti- Strep-Tag II Mo Ab (A-C) or with an anti-C4BP-a chain polyclonal Ab (D-F). The results shown are the mean ± SD from 3-6 independent experiments performed in duplicate (*, p < 0.05; **, p < 0.01).

[0016] Figure 6.- C4BP(0-) but not PRP6-HO7 binds to LRP1. ELISA measurements of (A) immobilized hLRPl IV-Fc (A), or hLRPl II (B) binding to PRP6-HO7 or C4BP(0-). The results shown are the mean ± SD from 4 independent experiments performed in duplicate (*, p < 0.05).

[0017] Figure 7.- C4BP(0-) and PRP6-HO7 modulation of CD36 expression by differentiating Mo-macrophages. Time-course of CD36 transcript (A) and surface CD36 expression (B) in differentiating Mo-macrophages, either untreated (MO), C4BP(0+)-, C4BP(0-), or PRP6-HO7-treated. Data are given as (A) relative log2FC respect to the CD36 transcript expression of day 0 (monocytes), and (B) median fluorescence intensity (MFI). The results shown are the mean ± SD from 4 (CD 36 transcript analysis) or 7 (surface CD36 analysis) independent donor samples (**, p < 0.01; ***, p < 0.001; ****, p < 0.0001 compared with M0).

[0018] Figure 8.- PRP6-HO7 and C4BP(0-) prevent oxLDL uptake by differentiating Mo- macrophages. Differentiating Mo-macrophages were either untreated or treated with PRP6-HO7, anti-hCD36 MoAb, the isotype control anti-hlgGlK MoAb (all at 32 nM), or SSO (200 pM) for 16 h. The cells then were incubated with Dil-OxLDL (2.5 pg / ml) for 3 h and its uptake was analyzed by flow cytometry. The results shown are the mean MFI ± SD from 6 independent donor samples (*, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with untreated Mo-macrophages (M0 Dil-OxLDL)).

[0019] Figure 9.- Influence of C4BP variants regarding oxLDL uptake by differentiating Mo-macrophages. Differentiating Mo-macrophages were either untreated or treated with C4BP variants C4BP(0+), C4BP(0-) (both at 12 nM), PRP6-HO7, and PRP6-NO (both at 32 nM) for 16 h. The cells then were incubated with Dil-OxLDL (2.5 pg / ml) for 3 h and its uptake was analyzed by flow cytometry. The results shown are the mean MFI ± SD from 6 independent donor samples (*, p < 0.05 compared with untreated Mo- macrophages (M0 Dil-OxLDL)).

[0020] Figure 10.- CD36 overexpression in inflammatory conditions. (A) Surface CD36 expression in CD14+monocytes from IBD patients (n= 30) and healthy individuals (control) (n= 31) was assessed by flow cytometry. The results shown are the mean MFI ± SD (****, p < 0.0001). (B) Representative histograms of surface CD36 expression in the A375 melanoma cell line. Grey left histogram, background control; middle histogram, basal surface CD36 expression; right histogram, pro-inflammatory surface CD36 expression. (C)Median fluorescence intensity (MFI) quantification of basal surface CD36 expression (No induction) and pro-inflammatory (TNFa + IFNy) surface CD36 expression (n= 3).

[0021] Figure 11.- Comparative immunomodulatory activity of C4BP( -) and PRP6-HO7 versus different anti-human CD36 Ab isotypes. Human CD14+monocytes were incubated at the initial 24 h of their differentiation to Mo-DCs (iDC), or to Mo- macrophages (MO), with C4BP(P-), PRP6-HO7, or with different anti-human CD36 Ab isotypes (IgGlK, IgA, IgM) (all at 32 nM). DC maturation was achieved by LPS treatment, and Ml polarization was achieved by LPS + IFNy treatment. Cells were then collected, washed, and analyzed by flow cytometry for cell surface expression of CD83 and CD86 (mDCs), CD64 (MO), and CD80 (Ml). MFI, median fluorescence intensity for the different cell surface markers. iDC, untreated immature Mo-DCs; mDC, untreated LPS-matured Mo-DCs; MO, untreated Mo-macrophages; Ml, untreated classically polarized Mo-macrophages. Results shown are the mean + SD from 3 independent donor samples (*p < 0.05 compared with mDC, MO, or Ml, where appropriate).

[0022] Figure 12.- Comparative immunomodulatory activity of C4BP( -) and PRP6-HO7 versus different commercial biologies. Human CD14+monocytes were incubated at the initial 24 h of their differentiation to Mo-DCs (iDC), or to Mo-macrophages (MO), with C4BP(P-), PRP6-HO7, or with the commercial biologies Etanercept, Infliximab, and Abatacept (all at 32 nM). DC maturation was achieved by LPS treatment, and Ml polarization was achieved by LPS + IFNy treatment. Cells were then collected, washed, and analyzed by flow cytometry for cell surface expression of CD83 and CD86 (mDCs), CD64 (MO), and CD80 (Ml). The DC supernatants were collected, and secreted TNF-a and IL-6 levels were assessed by specific ELISAs. MFI, median fluorescence intensity for the different cell surface markers. iDC, untreated immature Mo-DCs; mDC, untreated LPS-matured Mo-DCs; MO, untreated Mo-macrophages; Ml, untreated classically polarized Mo-macrophages. Results shown are the mean + SD from 4-5 independent donor samples (*p < 0.05; **p < 0.01 compared with mDC, M0, or Ml, where appropriate).

[0023] Figure 13.- C4BP(0-) mitigates steatosis in ApoE(- / -) mice feed with a high-fat diet. Representative histology images from liver sections of ApoE(- / -) mice fed with a high- fat diet and either untreated (PBS) or treated with C4BP(0-). Pronounced steatosis (lipid droplets) is evident in the livers of control (PBS) mice but absent in the livers of C4BP(0-)-treated mice. Upper panels show Hematoxylin & Eosin staining; lower panels show Oil Red O staining. Scale bar, 100 pm.

[0024] Figure 14.- CD36 expression on CD14+human monocytes from a type I CD36- deficient (P36 1) and two normal (ST13 and ST14) individuals. Surface CD36 expression on CD14+monocytes was assessed by flow cytometry. Shown are representative histograms from a type I CD36-deficient (P36 1) (A) and two normal (STB and ST14) individuals (B and C). Grey histograms, background staining from the IgGl isotype control; white histograms, basal surface CD36 expression. The mean and median fluorescence intensity quantification is indicated at the bottom of the histogram images.

[0025] Figure 15.- C4BP( -) and PRP6-HO7 are unable to modulate inflammatory cytokine / chemokine secretion in CD36-deficient Mo-DCs. The concentrations of the indicated cytokines / chemokines in the supernatants from untreated, immature DCs (iDC), or LPS-matured DCs (mDCs), were compared with those from CBP(0+)-, C4BP(0-)-, or PRP6-HO7-treated, and LPS-matured DCs. Upper graphs, analysis of the P36 1 CD36-deficient individual (CD36def). Lower graphs, analysis of a representative normal individual (WT). Shown are the means of two technical replicates from each measurement.

[0026] Figure 16.- Therapeutic efficacy of PRP6-HO7 in the acute DSS-induced rat colitis model. A) Experimental design and treatment schedule of the DSS-induced rat acute colitis model. Except in the blank group, colitis was induced by administration of 5% DSS dissolved in water for 7 days and then switched to normal water. The experimental group was subcutaneously administered PRP6-HO7 on day 5. The reference group underwent daily mesalazine administration by oral gavage. B) PRP6-HO7 effectively reduced the clinical signs of acute DSS-induced colitis in rats. Variation of body weight in percentage vs. day 0. C) Disease Activity Index (DAI) evolution. D) DAI Area Under the Curve (AUC) evolution. Data are presented as mean ± SEM for each group (n=8 / group) (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 compared with DSS-induced and untreated rats). The blank group significance (Fig 16B and 16C) has not been included in the graphs. Figure 17.- Comparison of colon sizes in the acute DSS-induced rat colitis model. A) Macroscopic appearance of colons; image representative from each group. Colon length was measured as an indirect marker of inflammation. B) Colon weight (normalized by body weight) / length ratio. Values are the mean ± SEM for each group; (*, p < 0.05; **, p < 0.01 compared with DSS-induced and untreated rats).

[0027] Figure 18.- PRP6-HO7 attenuates colonic damage and histopathological signs in acute DSS-induced rat colitis. A) Representative sections of colon tissue were stained with H&E. High resolution cropped images of colon epithelia from each treatment group (scale bar: 100 pm). Whole transversal sections from representative colons for each treatment group obtained through tiled images (scale bar: 200 pm). Immune inflammatory cell infiltration and / or epithelial erosion and crypt loss, along with a hyperplasic and disorganized intestinal epithelial barrier was evident in untreated and mesalazine-treated DSS-induced rats. In contrast, the intestinal epithelia of the PRP6- HO7-treated DSS-induced rats resembled that of a normal intestinal epithelium from uninduced rats (Blank). B) Histopathological score of colon tissues from rats given water alone (Blank), DSS, or DSS plus PRP6-HO7 or Mesalazine. Data are presented as mean ± SEM (n=4 / group) (**, p < 0.01; ***, p < 0.001 compared with DSS-induced and untreated rats).

[0028] Figure 19.- PRP6-HO7 reduces colonic lamina propria immune cell infiltration in acute DSS-induced rat colitis. A) Flow cytometry analysis of colonic lamina propria immune cells (CDl lb+ GR1+) from a representative individual belonging to the untreated and DSS-induced group (DSS), and B) from a representative individual belonging to the PRP6-HO7-treated and DSS-induced group (PRP6-HO7). Dot plot images display the presence of a significant inflammatory immune cell infiltrate only in the DSS group (41%), and not in the PRP6-HO7 group (1 %).

[0029] DETAILED DESCRIPTION OF THE INVENTION

[0030] The authors of the present invention have surprisingly found that PRP6-HO7 and C4BP(P~) bind with high affinity to CD36. Under steady state conditions, CD36 is not expressed or is present at low density in the cell surface, in which case C4BP(0-) and PRP6-HO7 would not affect CD36 function. Nevertheless, in pro-inflammatory conditions, CD36 would be overexpressed, and its cell surface density would increase to the point to allow C4BP(0-) or PRP6-HO7 intervention through CD36 clustering. PRP6-HO7 interaction with the cell surface down-regulates CD36 expression, probably through PRP6-HO7-CD36 internalization. Moreover, PRP6-HO7 and C4BP(P~) interaction with CD36 is very specific, because PRP6-HO7 does not interact with other known C4BP(0-) ligands such as LRP-1 (Western et al. (2002) J. Biol. Chem. 277: 2511-6), or the closest CD36 homologue SR-B1 (CD36L1). Thus, given the key relevance of CD36 in a variety of pathological processes and the unique mechanism of action of C4BP(0-) and PRP6-HO7, the authors have demonstrated its therapeutic potential in various pathologies.

[0031] In a first aspect, the invention relates to an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0032] Alternatively, the invention relates to a method for preventing and / or treating a disease involving an undesired activation of the immune system which comprises administering an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, to a subject having CD36 expression on the surface of the monocytes / macrophages.

[0033] Alternatively, the invention relates to the use of an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain for the preparation of a medicament for the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0034] In a second aspect, the invention relates to an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID- 19, hepatitis B and C, HIV-induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid-driven lung injury.

[0035] Alternatively, the invention relates to a method for preventing and / or treating a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury comprising administering to a subject in need thereof an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain.

[0036] Alternatively, the invention relates to the use of an oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain for the preparation of a medicament for the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, nonalcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic- induced gallstones, COVID-19, hepatitis B and C, HIV-induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid-driven lung injury.

[0037] “Oligomer” as used herein relates to a macromolecular complex formed by non- covalent bonding of proteins or polypeptides that consist of a few repeating units. Each of the repeating units is a monomer, a polypeptide comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain.

[0038] In a preferred embodiment, the oligomer comprises at least four polypeptides as previously defined. In another preferred embodiment, the oligomer comprises at least five polypeptides as previously defined. In another preferred embodiment, the oligomer comprises at least six polypeptides as previously defined. In another preferred embodiment, the oligomer comprises at least seven polypeptides as previously defined, or more preferably at least eight polypeptides as previously described.

[0039] In another preferred embodiment, the oligomer is a homooligomer. “Homooligomer” as used herein refers to an oligomer formed by identical monomers. Particularly, the homooligomer of the present invention is a hexamer or a heptamer, composed of six or seven monomers, respectively. In a preferred embodiment, at least one of the monomers of the homooligomers of the invention is a recombinant polypeptide according to the invention. In another preferred embodiment, all monomers of the homooligomers of the invention are recombinant polypeptides. The obtention of homooligomers involves, for example, creating a recombinant sequence encoding the CCP6 domain of C4BP alpha chain and the oligomerization domain in which the 3’ end of a DNA sequence encoding the CCP6 domain is ligated to the 5’ end of a DNA sequence encoding the oligomerization domain. Upon expression in an appropriate host, this hybrid DNA sequence will produce the recombinant polypeptide of the invention that will assemble into an oligomer, particularly into an homooligomer, more particularly into an heptamer formed by seven identical recombinant polypeptides. The homooligomers of the invention associate spontaneously in the recombinant expression system used. Methods to assess if a complex is an homooligomer of the invention is for example an homo-oligomer structure prediction from its monomer sequence / structure using the GalaxyHomomer server (http: / / galaxy.seoklab.org / homomer) (Baek et al. (2017) Nucleic Acids Res. 45: W320-W324).

[0040] The term “polypeptide” or “protein”, as used herein, refers to a polymer of amino acid residues covalently linked by peptide bonds forming a linear chain. The terminal amino acid at one end of the chain (amino terminal) has a free amino group, while the terminal amino acid at the other end of the chain (carboxy terminal) has a free carboxyl group. As used herein, the term “amino terminus” (abbreviated N-terminus) refers to the free amino group on an amino acid at the amino terminal of a polypeptide or to the amino group when participating in a peptide bond of an amino acid at any other location within the peptide. Similarly, the term “carboxy terminus” refers to the free carboxyl group on the carboxy terminus of a polypeptide or the carboxyl group of an amino acid at any other location within the peptide. In an embodiment the polypeptide of the invention has less than 1000 amino acids, less than 900 amino acids, less than 800 amino acids, less than 700 amino acids, less than 600 amino acids, less than 500 amino acids, less than 475 amino acids, less than 450 amino acids, less than 425 amino acids, less than 400 amino acids, less than 375 amino acids, less than 350 amino acids, less than 325 amino acids, less than 300 amino acids, less than 275 amino acids, less than 250 amino acids, less than 225 amino acids, less than 200 amino acids or less than 175 amino acids. In a preferred embodiment, the polypeptide of the invention has between 100 and 200 amino acids, preferably between 115 and 175 amino acids, more preferably between 119 and 173 amino acids, even more preferably between 119 and 125 amino acids. In a preferred embodiment, the polypeptide of the invention has 167 amino acids. In another preferred embodiment, the polypeptide of the invention has 119 amino acids.

[0041] In a preferred embodiment, the polypeptide forming the oligomer is a recombinant polypeptide. The expressions “recombinant polypeptide” or “recombinant protein”, are used herein interchangeably, and refer to a polypeptide or protein that fusions the CCP6 domain of C4BP alpha chain and an oligomerization domain that is obtained by recombinant DNA methods upon expression of a recombinant polynucleotide. “Recombinant”, as used herein, means that a particular nucleic acid (DNA or RNA) or vector is the product of various combinations of cloning, restriction, polymerase chain reaction (PCR) and ligation steps resulting in a construct having a structural coding sequence distinguishable from endogenous nucleic acids found in natural systems. DNA sequences encoding polypeptides can be assembled from cDNA fragments or from a series of synthetic oligonucleotides to provide a synthetic nucleic acid which is capable of being expressed from a recombinant transcriptional unit contained in a cell or in a cell-free transcription and translation system. Genomic DNA comprising the relevant sequences can also be used in the formation of a recombinant gene or transcriptional unit. Thus, the term “recombinant” nucleic acid refers to one which is not naturally occurring, e.g., is made by the artificial combination of two otherwise separated segments of sequence through human intervention. This artificial combination is often accomplished by either chemical synthesis means, or by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques, in order to join together nucleic acid segments of desired functions to generate a desired combination of functions.

[0042] As used herein, an "amino acid residue" refers to any naturally occurring amino acid, any amino acid derivative or any amino acid mimic known in the art. In certain embodiments, the residues of the polypeptide are sequential, without any non-amino acid interrupting the sequence of amino acid residues. In other embodiments, the sequence may comprise one or more non-amino acid moieties. In particular embodiments, the sequence of residues of the polypeptide may be interrupted by one or more non-amino acid moieties (see for example, Li et al. 2013. Molecules, 18(8):9797- 9817). The terms “C4BP” or “C4b-binding protein”, as used herein, refer to a regulatory component of the classical complement pathway that is mainly synthesized by liver cells which acts as a cofactor for Factor I-dependent degradation of C3b and C4b and accelerates the decay of classical complement pathway C3 / C5-convertases. C4BP circulates in the plasma as three isoforms, the proportion of which depends on the relative levels of C4BPa (70 kDa) and C4BP0 (45 kDa) chains. The major isoform of C4BP is composed of 7 identical a-chains and 1 P-chain (a?Pi) and is termed C4BP(a?Pi) or C4BP(P+). Upon inflammation a normally less abundant isoform is up- regulated that is exclusively composed of 7 a-chains (a?Po), dubbed C4BP(a?Po) or C4BP(P ). Moreover, recombinant expression of the a-chains in eukaryotic cells might result in an oligomer comprising 6 a-chains (a60o). In a preferred embodiment, the oligomer for use according to the invention is a C4BP isoform selected from the group consisting of a?Po and aePo.

[0043] The term “C4BP a-chain”, also known as PRP or proline-rich protein, as used herein, refers to the mature processed form of the human polypeptide defined under accession number P04003 in the NCBI database (release of November 8, 2023) and which comprises amino acids 49 to 597. The term C4BP a-chain is also used to refer to orthologs of the human C4BP a-chain such as the mouse C4BP a-chain corresponding to the mature form of the polypeptide shown in the NCBI database under accession number P08607 (release of November 8, 2023) (amino acids 57 to 469), the rat C4BP a- chain corresponding to the mature form of the polypeptide shown in the NCBI database under accession number Q63514 (release of November 8, 2023) (amino acids 14 to 558), the bovine C4BP a-chain corresponding to the mature form of the polypeptide shown in the NCBI database under accession number Q28065 (release of November 8, 2023) (amino acids 49 to 610).

[0044] The C4BP a-chain contains 8 complement control protein domains (CCP) which are 60 amino acid residues long with four cysteine residues disulfide bonded in a 1-3 2- 4 arrangement, and a hydrophobic core built around an almost invariant tryptophan residue. Thus, the term “CCP domain”, as used herein, refers to one of the complement control domains found in the C4BP alpha chain. In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises a full-length C4BP alpha chain. C4BP(0-) refers to the C4BP isoforms lacking the beta chain. In a preferred embodiment, the C4BP(0-) is a?Po, homooligomer consisting of 7 CB4P alpha chains.. In another preferred embodiment, the C4BP(0-) is aePo, hoomoligomer consisting of 6 CB4P alpha chains.

[0045] The expression “CCP6 domain”, as used herein, corresponds to the region found between amino acids 363 and 424 (SEQ ID NO: 1) with respect to the human C4BP alpha chain defined in the sequence provided in the NCBI database under accession number P04003 (release of November 8, 2023) and which corresponds to the sequence: LCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWSP RTPSCGD (SEQ ID NO: 1)

[0046] Thus, in a preferred embodiment, the CCP6 domain is the CCP6 domain of the human C4BP alpha chain, more preferably is SEQ ID NO: 1. In an embodiment, the CCP6 domain of the C4BP alpha chain comprises SEQ ID NO: 1. In another embodiment, the CCP6 domain of the C4BP alpha chain consists of SEQ ID NO: 1.

[0047] In another preferred embodiment, the functionally equivalent variant of CCP6 is a peptide having the sequence selected from the group consisting of SEQ ID NO: 2, 3, 4 and 5 or a functionally equivalent variant thereof (see Table I).

[0048] Table I. Peptides derived from CCP6 domain

[0049] In a preferred embodiment the peptide is SEQ ID NO: 5.

[0050] The term “peptide”, as used herein, relates to a linear chain of around 2 to 40 amino acids joined together with peptide bonds.

[0051] The term “CCP6 domain” is also used herein to refer to the CCP6 domain of the C4BP alpha chain of any ortholog of the human C4BP alpha chain such as, for example, the CCP6 domain of the rabbit C4BP alpha chain (amino acids 401-462) (SEQ ID NO: 12) with respect to the sequence provided in the NCBI database under accession number Z35490 (release of November 9, 2023), the CCP6 domain of the rat C4BP alpha chain (amino acids 327-388) (SEQ ID NO: 13) with respect to the sequence provided in the NCBI database under accession number Q63514 (release of November 8, 2023), or the CCP6 domain of the bovine C4BP alpha chain (amino acids 365-427) (SEQ ID NO: 14) with respect to the sequence provided in the NCBI database under accession number Q28065 (release of November 8, 2023) as set forth below in Table 2.

[0052] Table 2. CCP6 domain sequences of orthologs of human CCP6 domain of C4BP alpha chain.

[0053] The term “CCP6 domain” is also used herein to refer to any functionally equivalent variant of the naturally-occurring CCP6 domain defined above resulting from the substitution, insertion or deletion of one or more amino acids and which substantially preserves the ability to induce a tolerogenic phenotype of the original polypeptide when forming oligomers as shown in W02013 / 010998 A2.

[0054] In a preferred embodiment the functionally equivalent variant of the CCP6 domain results from modification of any of the above sequences by substitutions (e.g., conservative amino acid substitutions) and / or insertions (e.g., small, single amino acid insertions, or insertions encompassing 2, 3, 4, 5, 10, 15, 20, or more contiguous amino acids) and / or deletions (e.g., small, single amino acid deletions, or deletions encompassing 2, 3, 4, 5, 10, 15, 20, or more contiguous amino acids). Thus, in certain embodiments, a variant of a native sequence is one that differs from a naturally- occurring sequence by (i) one or more (e.g., 2, 3, 4, 5, 6, or more) conservative amino acid substitutions, (ii) deletion of one or more (e.g., 2, 3, 4, 5, 6, or more) amino acids, (iii) insertion of one or more (e.g., 2, 3, 4, 5, 6, or more) amino acids, or (iv) a combination thereof. Deleted or inserted amino acids can be contiguous or noncontiguous.

[0055] In making such changes, the hydropathy index of amino acids is considered since it is known that certain amino acids may be substituted for other amino acids having a similar hydropathy index or score and result in a polypeptide with similar biological activity. For example, the relative hydropathic character of an amino acid residue affects the secondary and tertiary structure of the resultant polypeptide, which in turn defines the interaction of the polypeptide with other molecules, such as enzymes, substrates, receptors, antibodies, antigens, and the like. As outlined above, amino acid substitutions are generally based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions which take various of the foregoing characteristics into consideration are well known to those of skill in the art and are set forth below in Table 3.

[0056] Table 3. Amino acid substitutions

[0057] In an embodiment, the functionally equivalent variant comprises additions of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the N-terminal end, in the C-terminal end or in both ends of the sequence of the CCP6 domain. In an embodiment, the functionally equivalent variant comprises additions of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In another embodiment, the functionally equivalent variant comprises deletions of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the N- terminal end, in the C-terminal end or in both ends of the sequence of the CCP6 domain. In another embodiment, the functionally equivalent variant comprises deletions of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In another embodiment, the functionally equivalent variant has a substitution of at least one, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the sequence of the CCP6 domain. In another embodiment, the functionally equivalent variant has a substitution of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In a preferred embodiment, the functionally equivalent variant is a variant having the addition of one amino acid at the N-terminal end of the CCP6 domain, preferably having the addition of a methionine.

[0058] In another embodiment, the functionally equivalent variant of the CCP6 domain of the C4BP alpha chain includes one or more of the residues equivalent to cysteine at position 2 of SEQ ID NO: 1, the cysteine at position 3 of SEQ ID NO: 1, the proline at position 4 of SEQ ID NO: 1, the proline at position 6 of SEQ ID NO: 1, the isoleucine at position 13 of SEQ ID NO: 1, the histidine at position 16 of SEQ ID NO: 1, the cysteine at position 25 of SEQ ID NO: 1, the tyrosine at position 27 of SEQ ID NO: 1, the glycine at position 30 of SEQ ID NO: 1, the aspartic acid at position 31 of SEQ ID NO: 1, the cysteine at position 37 of SEQ ID NO: 1, the cysteine at position 47 of SEQ ID NO: 1, the glycine at position 51 of SEQ ID NO: 1, the threonine at position 52 of SEQ ID NO: 1, the tryptophan at position 53 of SEQ ID NO: 1, the proline at position 55 of SEQ ID NO: 1, the threonine at position 57 of SEQ ID NO: 1, the proline at position 58 of SEQ ID NO: 1, and the cysteine at position 60 of SEQ ID NO: 1. Preferably, includes all of these residues. In a more preferred embodiment, the variant retains the relative spacing between these residues. Functionally equivalent variants of the CCP6 domain of the C4BP alpha chain include, without limitation, naturally occurring polymorphic variants (i.e., allelic variants) as well as recombinantly manipulated or engineered variants. CCP6 domain variants suitable for use according to the present invention include, without limitation, polypeptides having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, at least 50%, at least 48%, at least 45%, at least 40%, at least 35% sequence identity with the naturally- occurring CCP6 domains as defined above and, in particular, with the naturally- occurring CCP6 domain of human origin.

[0059] The percent identity of the amino acid sequence of a CCP6 domain variant to the amino acid sequence set forth above can be readily determined by persons skilled in the art by sequence comparison. As used herein, two amino acid sequences have 100 percent amino acid sequence identity if the amino acid residues of the two amino acid sequences are the same when aligned for maximal correspondence. Sequence comparisons of polypeptides and polynucleotides (for example, the polynucleotides that encode the polypeptides described herein) can be performed using any method such as those that use computer algorithms well known to persons having ordinary skill in the art. Such algorithms include Align or the BLAST algorithm (see, e.g., Altschul, J. Mol. Biol. 219:555-565, 1991; Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915- 10919, 1992), which are available at the NCBI website (see [online] Internet at ncbi.nlm.nih.gov / cgi-bin / BLAST). Default parameters may be used. In addition, standard software programs are available, such as those included in the LASERGENE bioinformatics computing suite (DNASTAR, Inc., Madison, Wis.); CLUSTALW program (Thompson et al., Nucleic Acids Res. 22:4673-80 (1991)); and "GeneDoc" (Nicholas et al., EMBNEW News 4: 14 (1991)). Other methods for comparing two amino acid sequences by determining optimal alignment are practiced by persons having skill in the art (see, for example, Peruski and Peruski, The Internet and the New Biology: Tools for Genomic and Molecular Research (ASM Press, Inc. 1997); Wu et al. (eds.), "Information Superhighway and Computer Databases of Nucleic Acids and Proteins," in Methods in Gene Biotechnology, pages 123-151 (CRC Press, Inc. 1997); and Bishop (ed.), Guide to Human Genome Computing, 2nd Ed. (Academic Press, Inc. 1998)).

[0060] The expression “substantially preserves the ability to induce a tolerogenic phenotype of the original polypeptide when forming oligomers”, as used herein, refers to polypeptides which are capable of inhibiting the maturation of dendritic cells and / or of macrophages as determined, e.g., in any of the examples 1 to 11 of the WO202 1074449 application when the CCP6 variant is fused to an oligomerization domain, preferably to the oligomerization domain of SEQ ID NO: 6. Particularly, the functionally equivalent variant shows an ability in generating tolerogenic dendritic cells when added to monocyte cells during the differentiation stage to immature dendritic cells and / or when added to immature dendritic cells during their maturation stage to mature dendritic cells. The ability of the variant to promote the generation of tolerogenic dendritic cells can be determined, e.g., by measuring the expression levels in the dendritic cells of maturation markers such as CD83, CD86 and / or CD80 of dendritic cells which have been matured in the presence of the variant (example 4 of the WO202 1074449 application). Thus, a polypeptide is considered as a functionally equivalent variant of the CCP6 domain of the C4BP alpha chain if it shows at least 100%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, or at least 50% of the activity of the CCP6 domain of the human C4BP alpha chain when it is fused to an oligomerization domain, particularly to the oligomerization domain of SEQ ID NO: 6.

[0061] The term “oligomerization domain”, as used herein, refers to a polypeptide domain having the property that polypeptides comprising said domain have a propensity to aggregate and form oligomers, i.e., is a region responsible for the oligomerization between monomers. The oligomerization domain contains amino acids in a region of one monomer that can interact with amino acids in a region of another monomer to enable oligomerization of the monomers. Suitable oligomerization domains are known in the art. In a preferred embodiment, the oligomerization domain is the oligomerization domain of C4BP alpha chain. In another embodiment, the oligomerization domain is a functionally equivalent variant of the oligomerization domain of the C4BP alpha chain. The expression “oligomerization domain of C4BP alpha chain”, as used herein, corresponds to the region found between amino acids 541 and 597 (SEQ ID NO: 6) with respect to the human C4BP alpha chain defined in the sequence provided in the NCBI database under accession number P04003 (release of November 8, 2023) and which corresponds to the sequence:

[0062] ETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRDSA RQSTLDKEL (SEQ ID NO: 6)

[0063] Therefore, in a preferred embodiment, the oligomerization domain is the oligomerization domain of human C4BP alpha chain or a functionally equivalent variant thereof; more preferably is SEQ ID NO: 6 or a functionally equivalent variant thereof; even more preferably is SEQ ID NO: 6. In an embodiment the oligomerization domain comprises SEQ ID NO: 6 or a functionally equivalent variant thereof, preferably SEQ ID NO: 6. In another embodiment, the oligomerization domain consists of SEQ ID NO: 6 or a functionally equivalent variant thereof, preferably SEQ ID NO: 6.

[0064] The term “oligomerization domain of C4BP alpha chain” is also used herein to refer to the oligomerization domain of any ortholog of the human C4BP alpha chain such as, for example, the oligomerization domain of the mouse C4BP alpha chain (amino acids 416-469) (SEQ ID NO: 15) with respect to the sequence provided in the NCBI database under accession number P08607 (release of November 8, 2023), the oligomerization domain of the rat C4BP alpha chain (amino acids 504-558) (SEQ ID NO: 16) with respect to the sequence provided in the NCBI database under accession number Q63514 (release of November 8, 2023), the oligomerization domain of the bovine C4BP alpha chain (amino acids (544-610) (SEQ ID NO: 17) with respect to the sequence provided in the NCBI database under accession number Q28065 (release of November 8, 2023) as set forth below in Table 4. The oligomerization domain of C4BP alpha chain of other mammalian orthologs useful in the present invention are SEQ ID NO: 18 to SEQ ID NO: 33 set forth in Table 4. Functionally equivalent variants of the oligomerization domain of human C4BP alpha chain useful in the present invention are SEQ ID NO: 34 to SEQ ID NO: 41. The oligomerization domain of avian ortho logs described in PCT patent application WO 2007 / 062819 are also included such as the oligomerization domain of chicken C4BP alpha chain (SEQ ID NO: 42) or its avian homologue in zebra finch (SEQ ID NO: 43). Functionally equivalent variants of the oligomerization domain of C4BP disclosed in table 1 of WO 2007 / 062819 are also included (SEQ ID NO: 44 to SEQ ID NO: 55).

[0065]

[0066] Table 4. Oligomerization domain sequences of orthologs and functionally equivalent variants of human oligomerization domain of C4BP alpha chain.

[0067] In a preferred embodiment, the oligomerization domain is the oligomerization domain of a mammalian C4BP alpha chain, more preferably of a human, mouse or rat C4BP alpha chain, most preferably of a human C4BP alpha chain.

[0068] In a preferred embodiment, the oligomerization domain is an oligomerization domain selected from the group consisting of SEQ ID NO: 6 and any one of SEQ ID NO: 15 to 55; preferably is selected from the group consisting of SEQ ID NO: 6 and any one of SEQ ID NO: 15 to SEQ ID NO: 41; more preferably is selected from the group consisting of SEQ ID NO: 6 and any one of SEQ ID NO: 15 to SEQ ID NO: 33; even more preferably is selected from the group consisting of SEQ ID NO: 6 and any one of SEQ ID NO: 15 to SEQ ID NO: 32; even more preferably is selected from the group consisting of SEQ ID NO: 6 and any one of SEQ ID NO: 15 to 17; even more preferably is SEQ ID NO: 6.

[0069] The oligomerization domain of C4BP alpha chain include also other mammalian and non-mammalian homologues of these sequences. The means to obtain such homologues are routine techniques available to those of skill in the art. In essence, such techniques include using nucleic acid encoding any of the sequences of oligomerization domains of the present invention, or fragments thereof, as a probe to recover and to determine the sequence of C4BP homologues in other species. A wide variety of techniques are available for this, for example PCR amplification and cloning of the homologue using a suitable source of mRNA (e.g. from an embryo or an actively dividing differentiated or tumor cell), or by methods comprising obtaining a cDNA library from the animal, e.g., a cDNA library from one of the above-mentioned sources, probing said library with a nucleic acid encoding any of the oligomerization domains of the present invention under stringent conditions, and recovering a cDNA encoding all or part of the sequences homologues of that animal. Where a partial cDNA is obtained, the full-length coding sequence may be determined by primer extension techniques. Alternatively, where all or part of the genome sequence of the animal is available, homology searches with the sequences of the invention may be used to determine suitable homologues.

[0070] The term “oligomerization domain of C4BP alpha chain” is also used herein to refer to any functionally equivalent variant of the naturally occurring oligomerization domain of C4BP alpha chain defined above resulting from the substitution, insertion or deletion of one or more amino acids and which substantially preserves the ability to form oligomers of the original polypeptide when forming part of a polypeptide of the invention.

[0071] Therefore, the functionally equivalent variant can be a fragment of the oligomerization domain wherein amino acids not relevant for the oligomerization ability have been deleted but amino acids relevant for the oligomerization are preserved, or a mutant wherein amino acids not involved in the oligomerization are mutated.

[0072] In a preferred embodiment, the functionally equivalent variant of the oligomerization domain of C4BP alpha chain results from modification of any of the above sequences by substitutions (e.g., conservative amino acid substitutions) and / or insertions (e.g., small, single amino acid insertions, or insertions encompassing 2, 3, 4, 5, 10, 15, 20, or more contiguous amino acids) and / or deletions (e.g., small, single amino acid deletions, or deletions encompassing 2, 3, 4, 5, 10, 15, 20, or more contiguous amino acids). Thus, in certain embodiments, a variant of a native sequence is one that differs from a naturally-occurring sequence by (i) one or more (e.g., 2, 3, 4,

[0073] 5, 6, or more) conservative amino acid substitutions, (ii) deletion of one or more (e.g., 2, 3, 4, 5, 6, or more) amino acids, (iii) insertion of one or more (e.g., 2, 3, 4, 5, 6, or more) amino acids, or (iv) a combination thereof. Deleted or inserted amino acids can be contiguous or non-contiguous.

[0074] In making such changes, the hydropathy index of amino acids is considered as was explained in relation to the functionally equivalent variant of CCP6 domain. Table 3 which discloses exemplary substitutions is also applicable to the functionally equivalent variants of the oligomerization domain of C4BP alpha chain.

[0075] In an embodiment, the functionally equivalent variant comprises additions of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the N-terminal end, in the C-terminal end or in both ends of the sequence of the oligomerization domain. In an embodiment, the functionally equivalent variant comprises additions of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In another embodiment, the functionally equivalent variant comprises deletions of at least 1, at least 2, at least 3, at least 4, at least 5, at least

[0076] 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the N- terminal end, in the C-terminal end or in both ends of the sequence of the oligomerization domain. In another embodiment, the functionally equivalent variant comprises deletions of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In another embodiment, the functionally equivalent variant has a substitution of at least one, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 amino acids in the sequence of the oligomerization domain. In another embodiment, the functionally equivalent variant has a substitution of less than 20, less than 15, less than 10, less than 9, less than 8, less than 7, less than 6, less than 5, less than 4, less than 3, less than 2 or 1 amino acids. In a preferred embodiment, the functionally equivalent variant is a variant having the addition of one amino acid at the N-terminal end of the oligomerization domain, preferably having the addition of a methionine.

[0077] In an embodiment, a fragment of the oligomerization domain of the C4BP alpha chain capable of forming oligomers may comprise at least 47 amino acids, preferably at least 50 amino acids.

[0078] In another embodiment, the variant of the oligomerization domain of the C4BP alpha chain includes one or more residues equivalent to glutamate at position 1 of SEQ ID NO: 6, glycine at position 12 of SEQ ID NO: 6, glutamine at position 17 of SEQ ID NO: 6, valine at position 25 of SEQ ID NO: 6, lysine at position 26 of SEQ ID NO: 6, alanine at position 28 of SEQ ID NO: 6, leucine at position 29 of SEQ ID NO: 6, glutamate at position 30 of SEQ ID NO: 6, tyrosine at position 32 of SEQ ID NO: 6, lysine at position 33 of SEQ ID NO: 6, leucine at position 34 of SEQ ID NO: 6, leucine at position 36 of SEQ ID NO: 6, glutamate at position 37 of SEQ ID NO:6, and leucine at position 41 of SEQ ID NO: 6. Preferably, the variant includes all of these residues. More preferably, the variant retains the relative spacing between these residues.

[0079] In a preferred embodiment, the variant includes the residues equivalent to glycine at position 12 of SEQ ID NO: 6, the alanine at position 28 of SEQ ID NO: 6, the leucines at positions 29, 34, 36 and 41 of SEQ ID NO: 6, the tyrosine at position 32 of SEQ ID NO: 2, and the lysine at position 33 of SEQ ID NO: 6. Preferably, the variant retains the relative spacing between these residues.

[0080] In another preferred embodiment, the variant of the oligomerization domain of the C4BP alpha chain includes the residues equivalent to glycine at position 12 of SEQ ID NO: 6, the alanine at position 28 of SEQ ID NO: 6, the leucines at positions 29, 34, 36 and 41 of SEQ ID NO: 6, the tyrosine at position 32 of SEQ ID NO: 6, the lysine at position 33 of SEQ ID NO: 6, and the two cysteine residues at positions 6 and 18 of SEQ ID NO: 6. Preferably, the variant retains the relative spacing between these residues.

[0081] Functionally equivalent variants of the oligomerization domain of the C4BP alpha chain for use according to the present invention include, without limitation, naturally occurring polymorphic variants (i.e., allelic variants) as well as recombinantly manipulated or engineered variants. Variants of the oligomerization domain suitable for use according to the present invention include, without limitation, polypeptides having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, at least 50%, at least 45%, at least 40%, at least 35% sequence identity with the naturally-occurring oligomerization domains as defined above and, in particular, with the naturally-occurring oligomerization domain of C4BP alpha chain of human origin.

[0082] Methods for determining the percent identity of the amino acid sequence of a variant of the oligomerization domain were previously described in the context of the CCP6 domain.

[0083] The expression “substantially preserves the ability to form oligomers of the original polypeptide when forming part of a recombinant polypeptide of the invention”, as used herein, refers to polypeptides which are capable of forming oligomers. Methods for determining whether a polypeptide is capable of forming oligomers are available to the skilled person and include, for instance, a method as described by Blom et al. (J.Biol.Chem. 2001, 276: 27136-27144) based on the analysis by polyacrylamide gel electrophoresis under native conditions of a purified C4BP obtained by recombinant expression of a variant a-chain in eukaryotic cell (e.g. 293 cells) followed by affinity purification using an antibody specific for CCP6 region. Alternatively, the ability of a variant of the oligomerization domain to form oligomers may be tested by expressing the variant in prokaryotic or eukaryotic host cells according to the invention and analyzing the variants recovered by polyacrylamide gel electrophoresis and Coomassie Blue staining under reducing and non-reducing conditions (i.e., by comparing the behaviour of the variant on an SDS-PAGE gel in the presence and absence of the reducing agent beta-mercaptoethanol) or by carrying out a Western blot probed with an anti-tag antibody or an anti-C4BP alpha chain antibody in reducing and non-reducing conditions as explained in Example 1 and Figure 3 of the WO2021074449 application. Alternatively, the ability of a variant of the oligomerization domain to form oligomers may be tested by expressing the variant in a prokaryotic host cell according to the invention, recovering the variant under conditions which result in oligomerization of the full 57 amino acid oligomerization domain of human C4BP alpha chain, and determining, e.g., by gel filtration, whether the variant forms oligomers.

[0084] Thus, a polypeptide is considered as a functionally equivalent variant of the oligomerization domain of C4BP alpha chain if it shows at least 100%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, or at least 50% of the capability of forming oligomers of the oligomerization domain of the human C4BP alpha chain when it is fused to a CCP6 domain, particularly to the CCP6 domain of SEQ ID NO: 1.

[0085] The functionally equivalent variant of the oligomerization domain of human C4BP alpha chain preferably maintains: (a) the cysteine residue equivalent to the cysteine residue 498 of the mature form of human C4BP alpha chain (i.e., the form of human C4BP alpha chain lacking the signal peptide), corresponding to amino acid 546 of the immature form defined under accession number P04003 in the NCBI database (release of November 8, 2023); (b) the cysteine residue equivalent to the cysteine residue 510 of the mature form of human C4BP alpha chain corresponding to amino acid 558 of the immature form defined under accession number P04003 in the NCBI database (release of November 8, 2023); and / or (c) the amphipatic a-helix region of the C-terminal end, more preferably the last 13 residues equivalent to residues 537-549 of the mature form of human C4BP alpha chain, corresponding to amino acids 585-597 of the immature form defined under accession number P04003 in the NCBI database (release of November 8, 2023). Preferably the functionally equivalent variant needs to maintain (c); more preferably the functionally equivalent variant needs to maintain (a), (b) and (c) for intracellular oligomerization of the molecule.

[0086] In a preferred embodiment of the oligomer for use according to the invention, the polypeptide does not comprise one or more of the domains selected from the group consisting of CCP1, CCP2, CCP3, CCP4, CCP5, CCP7 and CCP8 of the C4BP alpha chain. In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises a full length C4BP alpha chain or a functionally equivalent variant thereof. In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention is the full length C4BP alpha chain or a functionally equivalent variant thereof

[0087] When referring to human C4BP alpha chain, the CCP1 domain corresponds to amino acids 49 to 110, the CCP2 domain corresponds to amino acids 111 to 172, the CCP3 domain corresponds to amino acids 173 to 236, the CCP4 domain corresponds to amino acids 237 to 296, the CCP5 domain corresponds to amino acids 297 to 362, the CCP7 domain corresponds to amino acids 425 to 482 and the CCP8 domain corresponds to amino acids 483 to 540 of the polypeptide shown in the NCBI database under accession number P04003 (release of November 8, 2023). The person skilled in the art knows how to determine the CCPs equivalent to the human C4BP alpha chain in other C4BP alpha chains.

[0088] In an embodiment, the polypeptide consists of the CCP6 domain of the C4BP alpha chain and the oligomerization domain of the C4BP alpha chain; more preferably consists of the CCP6 domain of the human C4BP alpha chain and the oligomerization domain of the human C4BP alpha chain. Both domains can be in any order.

[0089] In a preferred embodiment, the polypeptide comprises the CCP6 domain of the C4BP alpha chain and the oligomerization domain of the C4BP alpha chain, pertain to the same species.

[0090] In another embodiment, the polypeptide consists of (a) a methionine, (b) the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof, and (c) an oligomerization domain; preferably consists of (a) a methionine, (b) the CCP6 domain of the C4BP alpha chain, and (c) the oligomerization domain of the C4BP alpha chain; more preferably consists of (a) a methionine, (b) the CCP6 domain of the human C4BP alpha chain, and (c) the oligomerization domain of the human C4BP alpha chain. The methionine is at the N-terminal end of the recombinant polypeptide and the domains can be in any order.

[0091] In a preferred embodiment, the polypeptide of the invention comprises SEQ ID NO: 7 or a functionally equivalent variant thereof, preferably comprises SEQ ID NO: 7. In a more preferred embodiment, the recombinant polypeptide of the invention consists of SEQ ID NO: 7 or a functionally equivalent variant thereof, preferably consists of SEQ ID NO: 7.

[0092] LCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWSP RTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRDS ARQSTLDKEL (SEQ ID NO: 7)

[0093] In another embodiment, the recombinant polypeptide of the invention consists of the sequence SEQ ID NO: 7 and an additional methionine at the N-terminal end of the polypeptide.

[0094] In an embodiment, the polypeptide of the invention does not comprise a region of a protein different from C4BP. For example, the polypeptide of the invention cannot be a fusion protein comprising a region which forms part of a different protein from C4BP.

[0095] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises a polypeptide which does not form part of C4BP. The polypeptide which does not form part of C4BP is considered as a heterologous polypeptide, i.e., a polypeptide that is derived from a different gene as that coding for C4BP.

[0096] The polypeptide of the invention may comprise the CCP6 domain or a functionally equivalent variant thereof and the oligomerization domain in any order. The oligomer for use according to the present invention may comprise a polypeptide comprising in an amino terminal to carboxy terminal direction, (a) the region which comprises the CCP6 domain of C4BP alpha chain, and (b) the region which comprises the oligomerization domain. Alternatively, the oligomer for use according to the present invention may comprise a polypeptide comprising in an amino terminal to carboxy terminal direction, (a) the region which comprises the oligomerization domain and (b) the region which comprises the CCP6 domain of the C4BP alpha chain. The person skilled in the art will understand that the oligomer for use according to the present invention may comprise a polypeptide that may contain additional amino acids between the CCP6 domain and the oligomerization domain. In a preferred embodiment, the C- terminus of the CCP6 domain of the C4BP alpha chain is directly fused to the N- terminus of the oligomerization domain, i.e., there is no additional amino acids or linkers between both domains. In another embodiment, the C-terminus of the oligomerization domain is directly fused to the N-terminus of the CCP6 domain of the C4BP alpha-chain. In another embodiment, the C-terminus of the CCP6 domain of the C4BP alpha chain is separated from the N-terminus of the oligomerization domain by additional amino acids, i.e., amino acids not present in the natural-occurring domains for which they derive. In another embodiment, the C-terminus of the oligomerization domain is separated from the N-terminus of the CCP6 domain of the C4BP alpha chain by additional amino acids. In a preferred embodiment, the polypeptide of the oligomer of the invention comprises, in an amino terminal to carboxy terminal direction, (a) a methionine, (b) the region which comprises the CCP6 domain of C4BP alpha chain, and (c) the region which comprises the oligomerization domain. In another embodiment, the polypeptide of the oligomer of the invention comprises, in an amino terminal to carboxy terminal direction, (a) a methionine, (b) the region which comprises the oligomerization domain and (c) the region which comprises the CCP6 domain of the C4BP alpha chain. In another embodiment, the recombinant polypeptide of the invention consists of an amino terminal to carboxy terminal direction, of (a) the CCP6 domain of C4BP alpha chain, and (b) the oligomerization domain of C4BP alpha chain. In another embodiment, the recombinant polypeptide of the invention consists of an amino terminal to carboxy terminal direction, of (a) the oligomerization domain of C4BP alpha chain, and (b) the CCP6 domain of C4BP alpha chain. All these embodiments apply equally when the polypeptide of the invention is formed by functionally equivalent variants of the CCP6 domain of the C4BP alpha chain and / or functionally equivalent variants of the oligomerization domain. All these embodiments, apply equally to one, or to two or to the at least three polypeptides of the oligomer for use according to the invention, preferably to four polypeptides of the invention, more preferably to five polypeptides of the invention, preferably to six polypeptides of the invention, and preferably to seven polypeptides of the invention.

[0097] The polypeptide forming part of the oligomer for use according to the invention can be in the form of a precursor. The term “precursor” refers to a polypeptide which, once processed, can give rise to the mature form of the polypeptide. The precursor is a polypeptide comprising a signal peptide to export the polypeptide to the extracellular media when the host cell is an eukaryotic cell.

[0098] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention further comprises a signal peptide. The expression “signal peptide”, as used herein, refers to a signal sequence which is located in the N-terminus of the polypeptide. As it is used herein, the term “signal peptide” or “signal sequence” or “secretory signal peptide” or “secretory signal sequence” refers to a peptide of a relatively short length, generally between 5 and 30 amino acid residues, directing proteins synthesized in the cell towards the secretory pathway. The signal peptide usually contains a series of hydrophobic amino acids adopting a secondary alpha helix structure. Additionally, many peptides include a series of positively-charged amino acids that can contribute to the protein adopting the suitable topology for its translocation. The signal peptide tends to have at its carboxyl end a motif for recognition by a peptidase, which is capable of hydrolyzing the signal peptide giving rise to a free signal peptide and a mature protein. The signal peptide can be cleaved once the protein of interest has reached the appropriate location. Any signal peptide may be used in the present invention. The signal sequence may be from the same species of the organism that is transformed or may be from a different species. As a way of illustrative, non-limitative examples, signal peptides from Chlamydomonas reinhardtii carbonic anhydrase (CAH1), signal peptide from Chlamydomonas reinhardtii periplasmic arylsulfatase (ARS1) or the signal peptide from Chlamydomonas reinhardtii Gametolysin Mi l may be used. Signal peptides useful for expression in bacteria can be Sec- or Tat- signal peptides. Signal peptides particularly for expression in yeasts can be the signal peptide from S.cerevisiae alpha-mating factor prepro peptide, the signal peptides from the P.pastoris acid phosphatase gene (PHO1) and the extracellular protein X (EPX1). In a preferred embodiment, the signal peptide is the signal peptide from alpha-factor mating peptide, preferably the alpha-factor mating peptide from Saccharomyces cerevisiae. Signal peptides useful for expression in mammalian eukaryotic cells are, without limitation, signal peptides from human OSM, VSV-G, mouse Ig Kappa, human IgG2 H, BM40, secrecon, human IgKVIII, CD33, tPA, human chymotrypsinogen, human trypsinogen-2, human IL-2, gaussia luc, albumin (HSA), influenza haemagglutinin, human insulin or silkworm fibroin LC.

[0099] In a preferred embodiment, the signal peptide is a human signal peptide. In a preferred embodiment, the signal peptide is the signal peptide of C4BP alpha chain, preferably of human C4BP alpha chain, more preferably is SEQ ID NO: 56 or a functionally equivalent variant thereof; more preferably is SEQ ID NO: 56. MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLG (SEQ ID NO: 56)

[0100] Therefore, in an embodiment, the signal peptide comprises SEQ ID NO: 56. In another embodiment the signal peptide consists of SEQ ID NO: 56.

[0101] Fusion of signal peptide to the polypeptide results in secretion of the fusion protein to the media, which is the preferred strategy since it permits easy and efficient purification from the extracellular medium.

[0102] In an embodiment, the oligomer for use according to the invention, comprises a polypeptide which is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) the region which comprises the CCP6 domain of C4BP alpha chain and (c) the region which comprises the oligomerization domain. Alternatively, the oligomer for use according to the invention, comprises a polypeptide which is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) the region which comprises the oligomerization domain and (c) the region which comprises the CCP6 domain of C4BP alpha chain. All these embodiments are equally applicable when the polypeptide consists of a signal peptide, a region consisting of the CCP6 domain of C4BP alpha chain, and a region consisting of the oligomerization domain. All these embodiments apply equally when the polypeptide of the invention is formed by functionally equivalent variants of the CCP6 domain of the C4BP alpha chain and / or functionally equivalent variants of the oligomerization domain.

[0103] In an embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises SEQ ID NO: 8 or a functionally equivalent variant thereof, preferably comprises SEQ ID NO: 8. In another embodiment the polypeptide of the invention consists of SEQ ID NO: 8 or a functionally equivalent variant thereof, preferably consists of SEQ ID NO: 8.

[0104] MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLGLCCPE PKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWSPRTPSC GDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRDSARQS TLDKEL (SEQ ID NO: 8)

[0105] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention does not comprise a signal peptide. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention further comprises a peptide that is not part of the C4BP alpha chain.

[0106] In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises a region which comprises the CCP6 domain of C4BP alpha chain, a region which comprises an oligomerization domain and one or more regions that comprise sequences which does not form part of C4BP. Any of these regions can be in any order with respect to each other.

[0107] Examples of polypeptides that comprise a peptide that is not part of the C4BP alpha chain can be, for example, fusion proteins that improve pharmacokinetic properties.

[0108] The polypeptide forming part of the oligomer for use according to the invention may be modified in order to modulate affinity for the receptor, modulate circulating half-life, modulate therapeutic half-life, modulate stability of the polypeptide, modulate cleavage by proteases, modulate dose, modulate release or bio-availability, facilitate purification, or improve or alter a particular route of administration. Similarly, the polypeptide may comprise protease cleavage sequences, reactive groups, or other molecules that improve detection, purification or other traits of the polypeptide. Preferably, the polypeptide comprises MBP sequences or AFV, slyD, tsf, SUMO, Bia, or GST sequences that are cleaved by proteases.

[0109] In a preferred embodiment, the peptide that is not part of the C4BP alpha chain is a tag peptide.

[0110] The expression “tag peptide”, as used herein, refers to a peptide suitable for detection, isolation and / or purification of the polypeptide. Non-limiting examples of tags include an affinity purification tag such as polyhistidine [poly(His)] sequences; peptide sequences capable of being recognized by antibodies that may be used to purify the resultant fusion protein by immunoaffinity chromatography, for example epitopes derived from the hemagglutinin of the fever virus, c-myc-tag (recognized by an anti-c- myc antibody); streptavidin-binding peptide tag or SBP-tag; S-tag; calmodulin-binding peptide (CBP); cellulose-binding domain; chitin-binding domain (CBD); glutathione S- transferase-tag; maltose-binding protein (MBP); 3xHA tag or hemagglutinin tag; NusA; TrxA; DsbA; Avi-tag; Strep-tag; an arginine tag (Arg-tag); FLAG-tag, etc. (Zhao et al. 2013. J. Anal. Methods Chemistry, 2013:581093; Terpe K. 2003. Appl. Microbiol. Biotechnol. 60(5): 523-533). Said affinity purification tag can be fused directly in-line or, alternatively, fused to the monomer polypeptide via a cleavable linker, i.e., a peptide segment containing an amino acid sequence that is specifically cleavable by enzymatic or chemical means (i.e., a recognition / cleavage site). In a particular embodiment, said cleavable linker comprises an amino acid sequence which is cleavable by a protease (or protease recognition site) once the protein has been translated. As illustrative, the cleavable linker can be an amino acid sequence cleavable by a protease such as enterokinase, Arg C endoprotease, Glu C endoprotease, Lys C endoprotease, factor Xa, thrombin, TEV (tobacco etch virus) protease, human rhinovirus 3C protease, sortase A, PreScission protease (Zhao et al. 2013. J. Anal. Methods Chem. 2013:581093), SUMO proteases (Butt TR. et al. 2005. Protein Expr. Purif. 43(1): 1-9), etc.; alternatively, in another particular embodiment, said cleavable linker comprises an amino acid sequence which is cleavable by a chemical reagent, such as, for example, cyanogen bromide which cleaves methionine residues, or any other suitable chemical reagent. In another embodiment, the cleavable linker is an intein. In a preferred embodiment, the cleavable linker is an amino acid sequence cleavable by enterokinase, preferably is the sequence DDDDK (SEQ ID NO: 57). The cleavable linker is useful if subsequent removal of the affinity purification tag is desirable. The tag may be located at any position of the monomer, particularly C-terminally or N-terminally to the CCP6 domain of C4BP alpha chain and to the oligomerization domain. In a more preferred embodiment, the tag peptide is linked to the N-terminal end of the CCP6 domain of C4BP alpha chain. In a preferred embodiment the tag is linked to the polypeptide via a linker cleavable by a TEV protease. In another embodiment the tag is linked to the polypeptide via a linker cleavable by enterokinase. In another embodiment, the tag is fused directly in-line with the polypeptide. In a more preferred embodiment, the tag is a poly(His) tag, preferably a poly(His) tag having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more than 20 histidines. In a preferred embodiment, the tag is a hexahistidine tag or His6-tag. In another embodiment, the poly(His) tag has at least 2 histidines, preferably at least 5 histidines, more preferably at least 6 histidines.

[0111] Preferably, the tag is a poly(His)tag, more preferably a His6-tag, and the tag is linked to the polypeptide via a linker cleavable by a TEV protease. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention of the invention comprises SEQ ID NO: 9 or a functionally equivalent variant thereof, preferably comprises SEQ ID NO: 9. In another embodiment the polypeptide forming part of the oligomer for use according of the invention consists of SEQ ID NO: 9 or a functionally equivalent variant thereof, preferably consists of SEQ ID NO: 9.

[0112] MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLG HHHHHHLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQG DGTWSPRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQ LELQRDSARQSTLDKEL (SEQ ID NO: 9).

[0113] The presence of any tags sequence in the fusion polypeptide of CCP6 and oligomerization domain of C4BP does not alter its immunomodulatory activity.

[0114] In another embodiment, the tag is a Strep-tag II peptide of SEQ ID NO: 58. WSHPQFEK (SEQ ID NO: 58).

[0115] The person skilled in the art will understand that the polypeptide forming part of the oligomer for use according to the invention can comprise more than one tag. Therefore, the polypeptide of the invention can have 1, 2, 3, 4, 5, 6 or more tags, preferably has 2 tags, more preferably has one tag. In a preferred embodiment, the polypeptide has a poly(His)tag and a Strep-tag II.

[0116] In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises a region which comprises the CCP6 domain of C4BP alpha chain, a region which comprises an oligomerization domain and one or more tag peptides. Any of these regions can be in any order with respect to each other. Thus, in an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) at least a tag peptide, (b) the region which comprises the CCP6 domain of C4BP alpha chain, and (c) the region which comprises the oligomerization domain. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) the region which comprises the CCP6 domain of C4BP alpha chain, (b) the region which comprises the oligomerization domain, and (c) at least a tag peptide. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) the region which comprises the CCP6 domain of C4BP alpha chain, (b) at least a tag peptide, and (c) the region which comprises the oligomerization domain. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises in an amino terminal to carboxy terminal direction (a) at least a tag peptide, (b) the region which comprises the oligomerization domain, and (c) the region which comprises the CCP6 domain of C4BP alpha chain. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction (a) the region which comprises the oligomerization domain, and (b) the region which comprises the CCP6 domain of C4BP alpha chain, and (c) at least a tag peptide. In another embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) the region which comprises the oligomerization domain, (b) at least a tag peptide, and (c) the region which comprises the CCP6 domain of C4BP alpha chain. The person skilled in the art will understand that, as used herein, the reference to “a tag peptide” refers both to a tag containing a cleavable linker or directly fused to the recombinant polypeptide. The person skilled in the art will understand that the expression “at least a tag peptide” means that the tag peptide of the recombinant polypeptide can be 1, 2, 3, 4, 5, 6 or more tag peptides, preferably 2 tag peptides, more preferably 1 tag peptides. The person skilled in the art will understand that all these embodiments may comprise an additional methionine at the N-terminal end of the polypeptide. All these embodiments are equally applicable when the polypeptide forming part of the oligomer for use according to the invention consists of at least a tag peptide, a region consisting of the CCP6 domain of C4BP alpha chain, and a region consisting of the oligomerization domain. All these embodiments apply equally when the polypeptide forming part of the oligomer for use according to the invention is formed by functionally equivalent variants of the CCP6 domain of the C4BP alpha chain and / or functionally equivalent variants of the oligomerization domain.

[0117] In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises a region which comprises the CCP6 domain of C4BP alpha chain, a region which comprises an oligomerization domain, at least a tag peptide and a signal peptide. Any of these regions can be in any order with respect to each other. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) at least a tag peptide, (c) a region which comprises the CCP6 domain of C4BP alpha chain, and (d) a region which comprises an oligomerization domain. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) at least a tag peptide, (c) a region which comprises an oligomerization domain, and (d) a region which comprises the CCP6 domain of C4BP alpha chain. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) a region which comprises an oligomerization domain, (c) at least a tag peptide, and (d) a region which comprises the CCP6 domain of C4BP alpha chain. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) a region which comprises the CCP6 domain of C4BP alpha chain, (c) at least a tag peptide, and (d) a region which comprises an oligomerization domain. In an embodiment, the polypeptide of the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) a region which comprises the CCP6 domain of C4BP alpha chain, (c) a region which comprises an oligomerization domain, and (d) at least a tag peptide. In an embodiment, the polypeptide forming part of the oligomer for use according to the invention is a fusion protein which comprises in an amino terminal to carboxy terminal direction, (a) a signal peptide, (b) a region which comprises an oligomerization domain, (c) a region which comprises the CCP6 domain of C4BP alpha chain, and (d) at least a tag peptide. The person skilled in the art will understand that, as used herein, the reference to “a tag peptide” refers both to a tag peptide containing a cleavable linker or directly fused to the recombinant polypeptide. The person skilled in the art will understand that the expression “at least a tag peptide” means that the tag peptide of the recombinant polypeptide can be 1, 2, 3, 4, 5, 6 or more tag peptides, preferably 2 tag peptides, more preferably 1 tag peptides. All these embodiments are equally applicable when the polypeptide of the invention consists of a signal peptide, at least a tag peptide, a region consisting of the CCP6 domain of C4BP alpha chain, and a region consisting of the oligomerization domain. All these embodiments apply equally when the polypeptide of the invention is formed by functionally equivalent variants of the CCP6 domain of the C4BP alpha chain and / or functionally equivalent variants of the oligomerization domain.

[0118] In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises SEQ ID NO: 10 or a functionally equivalent variant thereof, preferably comprises SEQ ID NO: 10. In a preferred embodiment the polypeptide forming part of the oligomer for use according to the invention consists of SEQ ID NO: 10 or a functionally equivalent variant thereof; more preferably consists of SEQ ID NO: 10.

[0119] HHHHHHLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQG DGTWSPRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQ LELQRDSARQSTLDKEL (SEQ ID NO: 10)

[0120] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of the sequence SEQ ID NO: 10 and an additional methionine at the N-terminal end of the polypeptide.

[0121] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises SEQ ID NO: 11 or a functionally equivalent variant thereof, preferably comprises SEQ ID NO: 11. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of SEQ ID NO: 11 or a functionally equivalent variant thereof; more preferably consists of SEQ ID NO: 11.

[0122] MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLGHHHH HHLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGT WSPRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLEL QRDSARQSTLDKEL (SEQ ID NO: 11)

[0123] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention does not comprise a tag peptide; preferably does not comprise a tag peptide and a signal peptide. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an aminoterminal to carboxy-terminal direction of (a) a signal peptide, (b) at least a tag peptide,

[0124] (c) a cleavable linker, (d) the CCP6 domain of the C4BP alpha chain, and (e) the oligomerization domain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction of (a) a signal peptide, (b) one tag peptide, (c) a cleavable linker, (d) the CCP6 domain of the C4BP alpha chain, and (e) the oligomerization domain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino terminal to carboxy-terminal direction of (a) the signal peptide of CB4P alpha chain, (b) a poly(His)tag, (c) a linker cleavable by TEV,

[0125] (d) the CCP6 domain of the C4BP alpha chain, and (e) the oligomerization domain of C4BP alpha chain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, SEQ ID NO: 59. MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLGHHHH HHENLYFQGLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAI CQGDGTWSPRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLE IEQLELQRDSARQSTLDKEL (SEQ ID NO: 59)

[0126] In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction of (a) at least a tag peptide, (b) a cleavable linker, (c) the CCP6 domain of the C4BP alpha chain, and (d) the oligomerization domain. In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction of (a) one tag peptide, (b) a cleavable linker, (c) the CCP6 domain of the C4BP alpha chain, and (d) the oligomerization domain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction, of (a) a poly(His)tag, (b) a linker cleavable by TEV, (c) the CCP6 domain of the C4BP alpha chain, and (d) the oligomerization domain of C4BP alpha chain. The person skilled in the art will understand that all these embodiments are applicable to polypeptides having a methionine at the N-terminal end. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, SEQ ID NO: 60.

[0127] HHHHHHENLYFQGLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETS RFSAICQGDGTWSPRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVY KLSLEIEQLELQRDSARQSTLDKEL (SEQ ID NO: 60)

[0128] In another embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of the sequence SEQ ID NO: 60 and an additional methionine at the N-terminal end of the polypeptide.

[0129] In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction of (a) a glycine residue, (b) the CCP6 domain of the C4BP alpha chain, and (c) the oligomerization domain, preferably the oligomerization domain of C4BP alpha chain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, SEQ ID NO: 61.

[0130] GLCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWS PRTPSCGDETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRD SARQSTLDKEL (SEQ ID NO: 61)

[0131] In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an aminoterminal to carboxy-terminal direction of (a) a methionine or a signal peptide, (b) two tag peptides, (c) a cleavable linker, (d) the CCP6 domain of the C4BP alpha chain, and (e) the oligomerization domain. In a more preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises or consists of, preferably consists of, in an amino-terminal to carboxy-terminal direction of (a) a methionine or a signal peptide, (b) a His6-tag, (c) a Strep-tag II, (d) a linker cleavable by enterokinase, (e) the CCP6 domain of the human C4BP alpha chain, and (!) the oligomerization domain of human C4BP alpha chain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of, in an amino-terminal to carboxy-terminal direction, of (a) a methionine, (b) a His6-tag, (c) a Strep-tag II of SEQ ID NO: 58, (d) a linker cleavable by enterokinase of SEQ ID NO: 57, (e) the CCP6 domain of the human C4BP alpha chain, and (f) the oligomerization domain of human C4BP alpha chain. In another preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of, in an amino-terminal to carboxy-terminal direction, of (a) a signal peptide, (b) a His6-tag, (c) a Strep-tag II of SEQ ID NO: 58, (d) a linker cleavable by enterokinase of SEQ ID NO: 57, (e) the CCP6 domain of the human C4BP alpha chain, and (f) the oligomerization domain of human C4BP alpha chain. In a preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention comprises SEQ ID NO: 62. In a more preferred embodiment, the polypeptide forming part of the oligomer for use according to the invention consists of SEQ ID NO: 62.

[0132] MHHHHHHWSH PQFEKDDDDK LCCPEPKLNN GEITQHRKSR PANHCVYFYG DEISFSCHET SRFSAICQGD GTWSPRTPSC GDETPEGCEQ VLTGKRLMQC LPNPEDVKMA LEVYKLSLEI EQLELQRDSA RQSTLDKEL (SEQ ID NO: 62)

[0133] Preferably, the polypeptide forming part of the oligomer for use according to the invention has a size between 75 and 1000 amino acids, more preferably between 100 and 800 amino acids, more preferably between 100 and 600 amino acids, more preferably between 110 and 550 amino acids, even more preferably between 115 and 550 amino acids, even more preferably between 119 and 550 amino acids. In an embodiment, the recombinant polypeptide of the invention preferably has at least 90 amino acids, at least 100 amino acids, at least 110 amino acids, at least 115 amino acids, at least 119 amino acids, at least 120 amino acids, at least 130 amino acids, at least 150 amino acids. In a preferred embodiment, the recombinant polypeptide of the invention has a size between 115 and 650 amino acids, preferably between 119 and 550 amino acids.

[0134] In a preferred embodiment, when insertions are made, preferably the size of the polypeptide does not exceed the length of the wild type sequence of C4BP alpha chain by more than 20, preferably by no more than 15, more preferably by no more than 10, amino acids. Thus, for example, when modified by insertion the protein containing the CCP6 domain of human C4BP alpha chain will desirably be no more than 617 amino acids. The person skilled in the art will understand that the different domains of the polypeptide forming part of the oligomer for use according to of the invention may be joined by spacers. As disclosed herein a spacer is an insert connecting or linking peptide of suitable length and character. In general, said spacer acts as a hinge region between said domains, allowing them to move independently from one another while maintaining the three-dimensional form of the individual domains. In this sense, a preferred spacer would be a hinge region characterized by a structural ductility or flexibility allowing this movement. The length of the spacer can vary. Typically, the number of amino acids in the spacer is 100 or less amino acids, preferably 50 or less amino acids, more preferably 40 or less amino acids, still more preferably 30 or less amino acids, or even more preferably 20 or less amino acids.

[0135] Alternatively, a suitable spacer can be based on the sequence of 10 amino acid residues of the upper hinge region of murine IgG3; which has been used for the production of dimerized antibodies by means of a coiled coil (Pack P. and Pluckthum, A., 1992, Biochemistry 31: 1579-1584) and can be useful as a spacer peptide according to the present invention. It can also be a corresponding sequence of the upper hinge region of human IgG3 or other human Ig subclasses (IgGl, IgG2, IgG4, IgM and IgA). The sequences of human Igs are not expected to be immunogenic in human beings. Additional spacers that can be used in the instant invention include the peptides of the amino acid sequences GAP, AAA.

[0136] In a particular embodiment, said spacer is a peptide having structural flexibility (i.e., a flexible linking peptide or “flexible linker”). Generally, such linkers are a few amino acids in length, such as from 1 to 20, e.g., from 2 to 10 amino acids in length. These amino acids are selected from the group consisting of glycine, serine, alanine and threonine. In another particular embodiment, the flexible linker is a peptide containing repeats of amino acid residues, particularly Gly and Ser, or any other suitable repeats of amino acid residues. Virtually any flexible linker can be used as spacer according to the invention. One such linker is a (Glym-Ser)nlinker, where m and n are each independently from 1 to 4. These are used in the art to attach protein domains to each other. Thus, the first component may be linked to the second by such a linker.

[0137] In a preferred embodiment, the CCP6 domain of C4BP alpha chain and the oligomerization domain are directly linked. In another embodiment, they are linked through a flexible linker. In a more preferred embodiment, the flexible linker is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20 residues.

[0138] It will be understood that sometimes it will be necessary to incorporate a sequence of amino acids suitable for expression of the protein. This will include at least an N-terminal methionine. The N-terminal sequence may include a cleavage site for chemical or enzymatic removal of all or part of the sequence.

[0139] The polypeptide forming part of the oligomer for use according to the invention can be modified to include any of a variety of known chemical groups or molecules. Such modifications include, but are not limited to, glycosylation, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment to polyethylene glycol (e.g., PEGylation), covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent crosslinks, formation of cystine, formation of pyroglutamate, formylation, gamma carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, esterification, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, ubiquitination, modifications with fatty acids, transfer-RNA mediated addition of amino acids to proteins such as arginylation, etc. Analogues of an amino acid (including unnatural amino acids) and peptides with substituted linkages are also included.

[0140] The polypeptide forming part of the oligomer for use according to the invention can be obtained by recombinant DNA techniques known to the person skilled in the art. Briefly, the recombinant nucleic acid encoding the CCP6 domain of C4BP alpha chain and the oligomerization domain is expressed in eukaryotic or prokaryotic cells and the product is recovered.

[0141] It should be understood that expression of polypeptides often involves post- translational modifications, such as cleavage of the signal sequence and / or cleavage of the tag peptide. The person skilled in the art will understand that the polypeptide forming part of the oligomer for use according to the invention as a whole needs to substantially preserve the ability to induce a tolerogenic phenotype of the original CCP6 domain of C4BP alpha chain and the ability to form oligomers of the original oligomerization domain, particularly, the ability to induce a tolerogenic phenotype of the CCP6 domain of human C4BP alpha chain and the ability to form oligomers of the oligomerization domain of C4BP alpha chain, more particularly the ability to induce a tolerogenic phenotype and the ability to form oligomers.

[0142] All the embodiments referring to the CCP6 domain of the C4BP alpha chain or the oligomerization domain apply equally to functionally equivalent variants of the CCP6 domain of the C4BP alpha chain and to functionally equivalent variants of the oligomerization domain.

[0143] Therefore, in an embodiment, the polypeptides forming part of the oligomer for use according to the invention have an additional amino acid at its N-terminal end.

[0144] In an embodiment, the polypeptides forming part of the oligomer for use according to the invention does not comprise a signal peptide and a tag peptide.

[0145] The invention also relates to a polynucleotide encoding the polypeptide of the invention comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0146] The invention also relates to a polynucleotide encoding the polypeptide of the invention comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment ofa disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID- 19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury.

[0147] Alternatively, the invention relates to a method for preventing and / or treating a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages comprising administering a polynucleotide encoding a polypeptide comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain.

[0148] The invention also relates to a method for preventing and / or treating a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury comprising administering a polynucleotide encoding a polypeptide comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain.

[0149] The terms “polynucleotide”, “nucleic acid” and “nucleic acid molecule” are used interchangeably to refer to polymeric forms of nucleotides of any length. The polynucleotides may contain deoxyribonucleotides, ribonucleotides, and / or their analogues. Nucleotides may have any three-dimensional structure, and may perform any function, known or unknown. The term "polynucleotide" includes, for example, singlestranded, double-stranded and triple helical molecules, a gene or gene fragment, exons, introns, mRNA, tRNA, rRNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. In addition to a native nucleic acid molecule, a nucleic acid molecule of the present invention may also comprise modified nucleic acid molecules. As used herein, mRNA refers to an RNA that can be translated in a cell.

[0150] The polynucleotides for use according to the invention may further comprise a single promoter region regulating the transcription of the region encoding the polypeptides of the invention provided that said promoters are compatible with the cells in which the polypeptides are to be expressed. The polynucleotides encoding for the polypeptides for use according to the invention can be found isolated as such or forming part of vectors allowing the propagation of said polynucleotides in suitable host cells.

[0151] In a preferred embodiment, the polynucleotide for use according to the invention is an expression cassette. In a preferred embodiment, the expression cassette comprises from 5 ’ to 3 ’ in the same open reading frame a nucleotide sequence encoding the CCP6 domain of C4BP alpha chain and a nucleotide sequence encoding an oligomerization domain. As it is used herein, the term “open reading frame” or “ORF” means a length of nucleic acid, either DNA, cDNA or RNA, that comprises a translation start signal or initiation codon, such as an ATG or AUG, and a termination codon and can be potentially translated into a polypeptide sequence. Said DNA sequence does not contain any internal end codon and can generally be translated into a peptide.

[0152] In a preferred embodiment, the expression cassette comprises from 5’ to 3’ a nucleotide sequence encoding a signal peptide, a nucleotide sequence encoding a first tag, a nucleotide sequence encoding a selectable marker, a nucleotide sequence encoding a second tag, a nucleotide sequence encoding a CCP6 domain of C4BP alpha chain and a nucleotide sequence encoding an oligomerization domain. In another embodiment, the expression cassette comprises from 5’ to 3’ a nucleotide sequence encoding a signal peptide, a nucleotide sequence encoding a first tag, a nucleotide sequence encoding a second tag, a nucleotide sequence encoding a cleavable linker, a nucleotide sequence encoding a CCP6 domain of C4BP alpha chain and a nucleotide sequence encoding an oligomerization domain. In another embodiment, the expression cassette comprises from 5’ to 3’ a nucleotide sequence encoding a signal peptide, a nucleotide sequence encoding a tag, a nucleotide sequence encoding a cleavable linker, a nucleotide sequence encoding a CCP6 domain of C4BP alpha chain and a nucleotide sequence encoding an oligomerization domain. In another embodiment, the expression cassette comprises from 5’ to 3’ a nucleotide sequence encoding a signal peptide, a nucleotide sequence encoding a tag, a nucleotide sequence encoding a CCP6 domain of C4BP alpha chain and a nucleotide sequence encoding an oligomerization domain. In a more preferred embodiment, all elements are in the same open reading frame. In a still more preferred embodiment, all elements are under the operation control of a regulatory nucleotide sequence. In a preferred embodiment, the first tag is different from the second tag.

[0153] Methods for cloning the nucleotide sequences into a vector and for expressing and purifying the recombinant polypeptides of the invention are well known in the art.

[0154] The invention also relates to a vector comprising a polynucleotide encoding the polypeptide of the invention, comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages. Alternatively, the invention also relates to method for preventing and / or treating a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages comprising administering a vector comprising the polynucleotide as previously defined.

[0155] The invention also relates to a vector comprising a polynucleotide encoding the polypeptide of the invention, comprising the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2- targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID- 19, hepatitis B and C, HIV-induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid-driven lung injury. Alternatively, the invention relate to a method for preventing and / or treating a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, nonalcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic- induced gallstones, COVID-19, hepatitis B and C, HIV-induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid-driven lung injury comprising administering a vector comprising the polynucleotide as previously defined.

[0156] The choice of expression vector will depend upon the choice of host. Vectors suitable for the insertion of said polynucleotides are vectors derived from expression vectors in prokaryotes such as pUC18, pUC19, Bluescript and the derivatives thereof, pET and the derivatives thereof, mp!8, mp!9, pBR322, pMB9, ColEl, pCRl, RP4, phages and “shuttle” vectors such as pSA3 and pAT28, expression vectors in yeasts such as vectors of the type of 2 micron plasmids, integration plasmids, YEP vectors, pBGZa vector, centromere plasmids and the like, expression vectors in insect cells such as vectors of the pAC series and of the pVL, expression vectors in plants such as pIBI, pEarleyGate, pAVA, pCAMBIA, pGSA, pGWB, pMDC, pMY, pORE series and the like, and expression vectors in eukaryotic cells, including baculovirus suitable for transfecting insect cells using any commercially available baculovirus system. The vectors for eukaryotic cells include preferably viral vectors (adenoviruses, viruses associated to adenoviruses (AAV), retroviruses and, particularly, lentiviruses) as well as non-viral vectors such as pSilencer 4.1-CMV (Ambion), pcDNA3, pcDNA3.1 / hyg, pHMCV / Zeo, pCR3.1, pEFI / His, pIND / GS, pRc / HCMV2, pSV40 / Zeo2, pTRACER- HCMV, pUB6 / V5-His, pVAXl, pZeoSV2, pCI, pSVL and PKSV-10, pBPV-1, pML2d and pTDTl. In a preferred embodiment, the vector is pcDNA3.1.

[0157] The vectors may also comprise a reporter or marker gene which allows identifying those cells that have been incorporated the vector after having been put in contact with it. Useful reporter genes in the context of the present invention include lacZ, luciferase, thymidine kinase, GFP and the like. Useful marker genes in the context of this invention include, for example, the neomycin resistance gene, conferring resistance to the aminoglycoside G418; the hygromycin phosphotransferase gene, conferring resistance to hygromycin; the ODC gene, conferring resistance to the inhibitor of the ornithine decarboxylase (2-(difluoromethyl)-DL-omithine (DFMO); the dihydrofolate reductase gene, conferring resistance to methotrexate; the puromycin-N- acetyl transferase gene, conferring resistance to puromycin; the ble gene, conferring resistance to zeocin; the adenosine deaminase gene, conferring resistance to 9-beta-D- xylofuranose adenine; the cytosine deaminase gene, allowing the cells to grow in the presence of N-(phosphonacetyl)-L-aspartate; thymidine kinase, allowing the cells to grow in the presence of aminopterin; the xanthine-guanine phosphoribosyltransferase gene, allowing the cells to grow in the presence of xanthine and the absence of guanine; the trpB gene of E. coli, allowing the cells to grow in the presence of indol instead of tryptophan; the hisD gene of E. coli, allowing the cells to use histidinol instead of histidine. The selection gene is incorporated into a plasmid that can additionally include a promoter suitable for the expression of said gene in eukaryotic cells (for example, the CMV or SV40 promoters), an optimized translation initiation site (for example, a site following the so-called Kozak’s rules or an IRES), a polyadenylation site such as, for example, the SV40 polyadenylation or phosphoglycerate kinase site, introns such as, for example, the beta-globulin gene intron. Alternatively, it is possible to use a combination of both the reporter gene and the marker gene simultaneously in the same vector. Vectors that contain both a promoter and a cloning site into which a polynucleotide for use according to the invention can be operatively linked are also provided in the present invention. Such vectors are capable of transcribing RNA in vitro or in vivo. In order to optimize expression and / or in vitro transcription, it may be necessary to remove, add or alter 5' and / or 3' untranslated portions of the clones to eliminate extra, potential inappropriate alternative translation initiation codons or other sequences that may interfere with or reduce expression, either at the level of transcription or translation. Alternatively, consensus ribosome binding sites can be inserted immediately 5' of the start codon to enhance expression. Gene delivery vehicles also include several non-viral vectors, including DNA / liposome complexes, and targeted viral protein-DNA complexes. Liposomes that also comprise a targeting antibody or fragment thereof can be used in the medical uses of this invention. To enhance delivery to a cell, nucleic acids or proteins of this invention can be conjugated to antibodies or binding fragments thereof which bind cell surface antigens.

[0158] As is well known in the art, for expression of the DNA sequences for use according to this invention, the DNA sequence should be operatively linked to an expression control sequence in an appropriate expression vector and employed in that expression vector to transform an appropriate unicellular host. Such operative linking of a DNA sequence of this invention to an expression control sequence, of course, includes the provision of a translation start signal in the correct reading frame upstream of the DNA sequence. If a particular DNA sequence being expressed does not begin with a methionine, the start signal will result in an additional amino acid (methionine) being located at the N-terminus of the product. While such a methionyl-containing product may be employed directly in the uses of the invention, it is usually more desirable to remove the methionine before use. Methods are known to those skilled in the art to remove such N-terminal methionine from polypeptides expressed with them. For example, certain hosts and fermentation conditions permit removal of substantially all of the N-terminal methionine in vivo. Other hosts require in vitro removal of the N- terminal methionine. However, such in vitro and in vivo methods are well known in the art.

[0159] The invention also relates to a host cell comprising the vector or the polynucleotide previously defined for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages. Alternatively, the invention relates to a method for preventing and / or treating a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages comprising administering a host cell comprising the vector or the polynucleotide previously defined to a subject in need thereof

[0160] The invention also relates to a host cell comprising the vector or the polynucleotide previously defined for use in the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury. Alternatively, the invention relates to a method for preventing and / or treating a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2- targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID- 19, hepatitis B and C, HIV-induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid-driven lung injury comprising administering a host cell comprising the vector or the polynucleotide previously defined to a subject in need thereof.

[0161] The term “host cell” is used such that it refers not only to the particular subject cell, but to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.

[0162] A wide variety of unicellular host cells are also useful in the present invention. In a preferred embodiment, the host cells are selected from bacteria, yeasts and mammalian eukaryotic cells. These hosts include well known eukaryotic and prokaryotic hosts, such as strains of E.coli, Pseudomonas, Bacillus, Lactobacillus, Thermophilus, Salmonella, Enterobacteriaceae or Streptomyces, fungi, such as yeasts, and animal cells, such as CHO and mouse cells, African green monkey cells, such as cos-1, COS-7, BSC 1, BSC 40, and BMT 10, insect cells, and human cells and plant cells in tissue culture. For animal cell expression, CHO, COS-7 cells and HEK293 cells are preferred, more preferably HEK293 cells.

[0163] For example, if E.coli from the genera Escherichia is used in the method of the invention, preferred strains of this bacterium to use would include BL21(DE3) and their derivatives including C41(DE3); C43(DE3)or C0214(DE3), C3030H, or other strains resistant to the toxicity of recombinant protein expression. Even more preferably, derivatives of these strains lacking the prophage DE3 may be used when the promoter is not the T7 promoter.

[0164] All the terms and embodiments previously described in the context of the polypeptide forming part of the oligomer for use according to the invention are equally applicable to the nucleotides, vectors and host cells for use according to the invention.

[0165] The oligomer, the polynucleotide, the vector or the host cell for use according to the invention, may be in the form of a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

[0166] Therapeutic methods of this invention comprise the step of treating patients in a pharmaceutically acceptable manner with those compositions. These compositions may be used to treat any mammal, including humans. The pharmaceutical compositions for use according to this invention may be in a variety of forms. These include, for example, solid, semi-solid and liquid dosage forms, such as tablets, pills, powders, liquid solutions or suspensions, liposomes, suppositories, injectable and infusible solutions and sustained release forms. Generally, the pharmaceutical compositions for use according to the present invention may be formulated and administered using methods and compositions similar to those used for pharmaceutically important polypeptide such as, for example, alpha interferon. Thus, the proteins for use in this invention may be stored in lyophilized form, reconstituted with sterile water just prior to administration, and administered by conventional routes of administration such as parenteral, subcutaneous, intravenous, intramuscular or intralesional routes. In a preferred embodiment, the compounds for use according to the invention are administered subcutaneously.

[0167] A composition for use according to the present invention may be a pharmaceutical composition that is a sterile aqueous or non-aqueous solution, suspension or emulsion, which additionally comprises a physiologically acceptable or suitable carrier. A pharmaceutically acceptable or suitable carrier may include (or refer to) an excipient (i.e., a non-toxic material that does not interfere with the activity of the active ingredient) and / or a diluent. Such compositions may be in the form of a solid, liquid, or gas (aerosol). Alternatively, compositions for use according to the invention may be formulated as a lyophilizate, or compounds may be encapsulated within liposomes using technology known in the art. Pharmaceutical compositions may also contain other components, which may be biologically active or inactive. Such components include, but are not limited to, buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, chelating agents such as EDTA or glutathione, stabilizers, dyes, flavouring agents, and suspending agents and / or preservatives.

[0168] Any suitable excipient or carrier known to those of ordinary skill in the art for use in pharmaceutical compositions may be employed in the compositions described herein. Excipients for therapeutic use are well known, and are described, for example, in Remingtons Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro ed. 1985). In general, the type of excipient is selected based on the mode of administration. Pharmaceutical compositions may be formulated for any appropriate manner of administration, including, for example, topical, oral, nasal, intrathecal, rectal, vaginal, intraocular, subconjunctival, sublingual or parenteral administration, including subcutaneous, intravenous, intramuscular, intrastemal, intracavemous, intrameatal or intraurethral injection or infusion. For parenteral administration, the carrier preferably comprises water, saline, alcohol, a fat, a wax or a buffer. For oral administration, any of the above excipients or a solid excipient or carrier, such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, kaolin, glycerine, starch dextrins, sodium alginate, carboxymethylcellulose, ethyl cellulose, glucose, sucrose and / or magnesium carbonate, may be employed.

[0169] A pharmaceutical composition (e.g., for oral administration or delivery by injection) may be in the form of a liquid. A liquid pharmaceutical composition may include, for example, one or more of the following: a sterile diluent such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils that may serve as the solvent or suspending medium, polyethylene glycols, glycerine, propylene glycol or other solvents; antibacterial agents; antioxidants; chelating agents; buffers and agents for the adjustment of tonicity such as sodium chloride or dextrose. A parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. The use of physiological saline is preferred, and an injectable pharmaceutical composition is preferably sterile.

[0170] The agents described herein, including polypeptides, homooligomers, polynucleotides, vectors, host cells, may be formulated for sustained or slow release. Such compositions may generally be prepared using well known technology and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Sustained-release formulations may contain an agent dispersed in a carrier matrix and / or contained within a reservoir surrounded by a rate controlling membrane. Excipients for use within such formulations are biocompatible, and may also be biodegradable; preferably the formulation provides a relatively constant level of active component release. The amount of active compound contained within a sustained release formulation depends upon the site of implantation, the rate and expected duration of release, and the nature of the condition to be treated or prevented.

[0171] Pharmaceutical compositions may be administered in a manner appropriate to the disease to be treated (or prevented) as determined by persons skilled in the medical art. An appropriate dose and a suitable duration and frequency of administration will be determined by such factors as the condition of the patient, the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. In general, an appropriate dose and treatment regimen provides the composition(s) in an amount sufficient to provide therapeutic and / or prophylactic benefit (e.g., an improved clinical outcome, such as more frequent complete or partial remissions, or longer disease-free and / or overall survival, or a lessening of symptom severity). For prophylactic use, a dose should be sufficient to prevent, delay the onset of, or diminish the severity of a disease associated with an immunological disease or disorder.

[0172] Optimal doses may generally be determined using experimental models and / or clinical trials. The optimal dose may depend upon the body mass, weight, or blood volume of the patient. In general, the amount of polypeptide present in a dose, or produced in situ by DNA present in a dose, ranges from about 0.01 pg to about 1000 pg per kg of host. The use of the minimum dosage that is sufficient to provide effective therapy is usually preferred. Patients may generally be monitored for therapeutic or prophylactic effectiveness using assays suitable for the condition being treated or prevented, which assays will be familiar to those having ordinary skill in the art. When administered in a liquid form, suitable dose sizes will vary with the size of the patient, but will typically range from about 1 ml to about 500 ml (comprising from about 0.01 pg to about 1000 pg per kg) for a 10-60 kg subject.

[0173] In a preferred embodiment, the pharmaceutical composition comprises from 0.45 mg to 18.90 mg of the homooligomer of the invention, more preferably from 0.45 mg to 9.45 mg, more preferably from 0.56 mg to 8.51 mg, more preferably from 0.75 mg to 8.13 mg, even more preferably from 0.79 mg to 8.05 mg. In a preferred embodiment, the dose is 0.79 mg. In another preferred embodiment, the dose is 0.8 mg. In another preferred embodiment, the dose is 8 mg. In another preferred embodiment, the dose is 8.05 mg. In an embodiment, the pharmaceutical composition comprises from 7.56 mg to 18.90 mg of a compound of the invention, preferably from 9.45 mg to 15.13 mg, preferably from 11.35 mg to 15.13 mg, preferably from 13.24 mg to 15.13 mg.

[0174] In an embodiment, the pharmaceutical composition comprises from 1.89 mg to 17.02 mg of a compound of the invention, preferably from 3.78 mg to 15.13 mg, preferably from 5.67 mg to 13.24 mg, more preferably from 7.56 mg to 11.35 mg.

[0175] In an embodiment, the pharmaceutical composition comprises from 0.5 mg to 9 mg of a compound of the invention, more preferably from 0.75 mg to 8.5 mg. In another embodiment, the composition comprises from 7 mg to 18.90 mg of a compound of the invention, more preferably from 7.5 mg to 18 mg, more preferably from 9 mg to 15 mg, more preferably from 11 mg to 15 mg, even more preferably from 13 mg to 15 mg. In another embodiment, the pharmaceutical composition comprises from 1 mg to 17 mg of a compound of the invention, preferably from 3 mg to 15 mg, preferably from 5 mg to 13 mg, more preferably from 7 mg to 11 mg.

[0176] For pharmaceutical compositions comprising an agent that is a nucleic acid molecule including an aptamer, the nucleic acid molecule may be present within any of a variety of delivery systems known to those of ordinary skill in the art, including nucleic acid, and bacterial, viral and mammalian expression systems such as, for example, recombinant expression constructs as provided herein. Techniques for incorporating DNA into such expression systems are well known to those of ordinary skill in the art. The DNA may also be "naked," as described, for example, in Ulmer et al., Science 259: 1745-49, 1993 and reviewed by Cohen, Science 259: 1691-1692, 1993. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells.

[0177] Nucleic acid molecules may be delivered into a cell according to any one of several methods described in the art (see, e.g., Akhtar et al., Trends Cell Bio. 2: 139 (1992); Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed. Akhtar, 1995, Maurer et al., Mol. Membr. Biol. 16: 129-40 (1999); Hofland and Huang, Handb. Exp. Pharmacol. 137: 165-92 (1999); Lee et al., ACS Symp. Ser. 752: 184-92 (2000); U.S. Pat. No. 6,395,713; International Patent Application Publication No. WO 94 / 02595); Selbo et al., Int. J. Cancer 87:853-59 (2000); Selbo et al., Tumour Biol. 23: 103-12 (2002); U.S. Patent Application Publication Nos. 2001 / 0007666, and 2003 / 077829). Such delivery methods known to persons having skill in the art, include, but are not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as biodegradable polymers; hydrogels; cyclodextrins (see, e.g., Gonzalez et al., Bioconjug. Chem. 10: 1068-74 (1999); Wang et al., International Application Publication Nos. WO 03 / 47518 and WO 03 / 46185); poly(lactic-co-glycolic)acid (PLGA) and PLCA microspheres (also useful for delivery of peptides and polypeptides and other substances) (see, e.g., U.S. Pat. No. 6,447,796; U.S. Patent Application Publication No. 2002 / 130430); biodegradable nanocapsules; and bioadhesive microspheres, or by proteinaceous vectors (International Application Publication No. WO 00 / 53722). In another embodiment, the nucleic acid molecules for use in altering (suppressing or enhancing) an immune response in an immune cell and for treating an immunological disease or disorder can also be formulated or complexed with polyethyleneimine and derivatives thereof, such as polyethyleneimine- polyethyleneglycol-N-acetylgalactosamine (PELPEG-GAL) or polyethyleneimine- polyethyleneglycol-tri-N-acetylgalactosamine (PEI-PEG-triGAL) derivatives (see also, e.g., U.S. Patent Application Publication No. 2003 / 0077829).

[0178] The pharmaceutical compositions / medicaments for use according to the invention may comprise further, e.g. active ingredients, e.g. other immunomodulatory antibodies such as anti-ICOS, anti-CD 154, anti-CD 134L or recombinant proteins such as, but not limited to rCTLA-4 (CD 152), rOX40 (CD134), or anti-inflammatory agents or immunomodulatory compounds such as, but not limited to cyclosporin A, FTY720, RAD, rapamycin, FK506, 15- deoxyspergualin, steroids; as described above.

[0179] In addition, one may use DNA sequences encoding recombinant polypeptides of the invention in somatic gene therapy. This involves, for example, inserting DNA sequences into retroviral-based vectors suitable for infection of human somatic cells (A. Kasid et al. 1990. “Human Gene Transfer: Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer”, Proc. Natl. Acad. Sci., USA, 87:473-477). For example, patients with an immunoinflammatory disease could be treated as follows. First, one would prepare a retrovirus characterized by a DNA sequence encoding a recombinant polypeptide of the invention. Then, one would isolate cells from the patient and infect them in vitro, with the retrovirus. One would then reintroduce these cells into the patient, where the vector will express and the cell will secrete the recombinant polypeptide of the invention which will assemble to form the oligomer for use according to the invention.

[0180] In another preferred embodiment, the compound of the invention is administered in a regimen comprising a plurality of administrations and wherein the compound is administered no more than once a week. In a more preferred embodiment, the compound of the invention is administered subcutaneously in a regimen comprising a plurality of administrations and wherein the compound is administered no more than once a week.

[0181] A plurality of administrations means at least two administrations. Therefore, in a preferred embodiment, the regimen comprises at least 2 administrations, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 30, at least 50, at least 100 or more. Preferably, the compound is administered chronically. Preferably, the compound is administered at least during 1 year, at least during 2 years, at least during 5 years or more.

[0182] The expression “the compound is administered no more than once a week” means that during a week the maximum number of administrations is one. This means that if the compound is administered on day 1, the subsequent administration cannot be on days 2, 3, 4, 5, 6 or 7. However, the expression “the compound is administered no more than once a week” encompasses the possibility that no administration is given during a week, for example, because the compound is administered once every two weeks. Therefore, according to the invention one administration is separated at least 7 days from another administration. Therefore, in a preferred embodiment, each administration is separated at least by 7 days from another, at least by 8 days, at least by 9 days, at least by 10 days, at least by 11 days, at least by 12 days, at least by 13 days, at least by 14 days, at least by 15 days, at least by 16 days, at least by 17 days, at least by 18 days, at least by 19 days, at least by 20 days, at least by 21 days, at least by 22 days, at least by 23 days, at least by 24 days, at least by 25 days, at least by 26 days, at least by 27 days, at least by 28 days, at least by 29 days, at least by 30 days, at least by 31 days, at least by 32 days, at least by 33 days, at least by 34 days, at least by 35 days, at least by 36 days, at least by 37 days, at least by 38 days, at least by 39 days, at least by 40 days, at least by 41 days, at least by 42 days, at least by 43 days, at least by 44 days, at least by 45 days, at least by 46 days, at least by 47 days, at least by 48 days, at least by 49 days, at least by 50 days, at least by 51 days, at least by 52 days, at least by 53 days, at least by 54 days, at least by 55 days, at least by 56 days, at least by 57 days, at least by 58 days, at least by 59 days, at least by 60 days or more.

[0183] In a preferred embodiment, the compound is administered once a week. In another preferred embodiment, the compound is administered once every two weeks. In another embodiment, the compound is administered once every three weeks. In another embodiment, the compound is administered once every four weeks. In another embodiment, the compound is administered once every five weeks. In another embodiment, the compound is administered once every six weeks. In another embodiment, the compound is administered once every seven weeks. In another embodiment, the compound is administered once every eight weeks. In another embodiment, the compound is administered once every nine weeks. In another embodiment, the compound is administered once every ten weeks. In another embodiment, the compound is administered once every eleven weeks. In another embodiment, the compound is administered once every twelve weeks. In an embodiment, the compound is administered monthly. In another embodiment, the compound is administered once every two months.

[0184] The compound of the invention has efficacy at low doses. Therefore, in a preferred embodiment, the dose of each administration ranges from 0.24 mg / m2to 9.99 mg / m2. In a more preferred embodiment, the dose of each administration ranges from 0.24 mg / m2to 5 mg / m2, preferably from 0.3 mg / m2to 4.5 mg / m2, preferably from 0.4 mg / m2to 4.3 mg / m2, even more preferably from 0.42 mg / m2to 4.26 mg / m2. In a preferred embodiment, the dose is 0.42 mg / m2. In another preferred embodiment, the dose is 4.26 mg / m2.

[0185] In an embodiment the dose of each administration ranges from 4 mg / m2to 9.99 mg / m2, preferably from 5 mg / m2to 8 mg / m2, preferably from 6 mg / m2to 8 mg / m2, preferably from 7 mg / m2to 8 mg / m2.

[0186] In an embodiment the dose of each administration ranges from 1 mg / m2to 9 mg / m2, preferably from 2 mg / m2to 8 mg / m2, preferably from 3 mg / m2to 7 mg / m2, more preferably from 4 mg / m2to 6 mg / m2. In another embodiment, the compound is administered at a dose of from 0.24 pg / m2to 0.24 mg / m2, preferably from 1 pg / m2to 0.24 mg / m2, preferably from 10 pg / m2to 0.24 mg / m2, preferably from 50 pg / m2to 0.24 mg / m2, more preferably from 100 pg / m2to 0.24 mg / m2, more preferably from 150 pg / m2to 0.24 mg / m2, more preferably from 200 pg / m2to 0.24 mg / m2.

[0187] As previously disclosed, the invention relates to medical uses for preventing and / or treating a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0188] “Subject”, as used herein relates to a human or may be a non-human primate or other animal (i.e., veterinary use). Examples of non-human primates and other animals include but are not limited to farm animals, pets, and zoo animals (e.g., horses, cows, buffalo, llamas, goats, rabbits, cats, dogs, chimpanzees, orangutans, gorillas, monkeys, elephants, bears, large cats, etc.). In a preferred embodiment, the compound is administered to a mammal, preferably a human.

[0189] In addition, the invention relate to medical uses for preventing and / or treating a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury.

[0190] The term “prevention”, as used herein, refers to the administration of a compound of the invention in an initial or early stage of the disease, or to also prevent its onset.

[0191] The term “treatment” is used to designate the administration of a compound of the invention to control the progression of the disease before or after the clinical signs have appeared. Control of the progression of the disease is understood as the beneficial or desired clinical results which include but are not limited to reduction of the symptoms, reduction of the duration of the disease, stabilization of pathological conditions (specifically avoiding additional impairment), delaying the progression of the disease, improving the pathological condition and remission (both partial and complete). The control of the progression of the disease also involves a prolongation of survival in comparison to the expected survival if the treatment was not applied.

[0192] The expression “disease involving an undesired activation of the immune system”, as used herein, refers to any disease which is involving an undesired activation of the immune system, including the innate or adaptative immune system as well as the humoral or cell branch of the immune system. Preferably, the immunological disease of the invention is a disease in which the immune system is activated in response to an alloantigen or an autoantigen. Therefore, immunological diseases in which the immune system is depressed are not encompassed by the present invention. In another embodiment, the disease is caused by an undesired activation of the immune system.

[0193] The subject to be prevented or treated of a disease involving an undesired activation of the immune system, is a subject having CD36 expression on the surface of the monocytes / macrophages. In a more preferred embodiment, the subject is a subject not having type I CD36 deficiency.

[0194] “Monocytes”, as used herein relates to a type of leukocyte or white blood cell. They are the largest type of leukocyte in blood and can differentiate into macrophages and monocyte-derived dendritic cells. Monocytes are produced by the bone marrow from precursors called monoblasts, bipotent cells that differentiated from hematopoietic stem cells. Monocytes circulate in the bloodstream for about one to three days and then typically migrate into tissues throughout the body where they differentiate into macrophages and dendritic cells.

[0195] “Macrophage” used herein, refers to a mononuclear phagocytic cell which is a type of white blood cell of the immune system that engulfs and digests cellular debris, foreign substances, microbes, cancer cells, and others in a process called phagocytosis. These large phagocytes are found in essentially all tissues. They take various forms with various names throughout the body (e.g., histiocytes, Kupffer cells, alveolar macrophages, microglia, and others), but all are part of the mononuclear phagocyte system. The term macrophage, as used herein, encompasses any of the types of macrophages known. Each type of macrophage, determined by its location, has a specific name. Specifically, the macrophages include, for example, adipose tissue macrophages, Kupffer cells (located in the liver), sinus histiocytes (located in the lymph nodes), alveolar macrophages (located in the pulmonary alveoli of lungs), tissue macrophages (histiocytes) leading to giant cells (located in the connective tissue), microglia (located in the central nervous system), Hofbauer cells (located in the placenta), intraglomerular mesangial cells (located in kidneys), osteoclasts (located in bones), epithelioid cells (located in granulomas), red pulp macrophages (located in red pulp of spleen), white-pulp macrophages (located in white-pulp of spleen), marginalzone macrophages and metallophilic macrophages (located in zones of the spleen), peritoneal macrophages (located in the peritoneal cavity), and LysoMac (located in Peyer’s patch). Besides phagocytosis, they play a critical role in nonspecific defence (innate immunity) and also help initiate specific defence mechanisms (adaptative immunity) by recruiting other immune cells such as lymphocytes. Macrophages also play an important anti-inflammatory role and can decrease immune reactions through the release of cytokines. There are a large number of commonly used macrophage markers such as CD14, CD16, CD64, CD68, CD71 and CCR5; the exact marker to be used will be dependent upon the subset of macrophage and the conditions of their local environment. Mature Ml macrophages are typically identified by markers CD86, CD80, CD68, MHC II, IL-1R, TLR2, TLR4, iNOS and / or SOCS3. M2 macrophages, particularly M2 macrophages activated by IL-4, are typically identified by markers CD163, MHC II, SR, MMR / CD206, CD200R, TGM2, DecoyR and / or IL-1R II. Precursors of macrophages include those having the phenotype IL3Ralow CD 11b- CD34+ C-KIT+ FLT3+ and IL3Rahigh CD 11b- CD34+ C-KIT+ FLT3+ (Xiao et al. (2015) Stem Cell Reports 4: 984-994).

[0196] “CD36” alternatively known as platelet membrane glycoprotein IV (GPIV), GPIIIb, thrombospondin receptor, collagen receptor, fatty acid translocase (FAT), and scavenger receptor class B, member 3 (SR-B3) is an integral membrane glycoprotein that has multiple physiological functions (Febbraio et al. (2001) J. Clin. Invest. 108: 785-91; Silverstein and Febbraio (2009) Sci. Signal. 2: re3). CD36 in humans corresponds to the sequence of reference Pl 6671 in the Uniprot database dated 8 November 2023. CD36 is a multiligand pattern recognition receptor (PRR) that interacts with many structurally dissimilar ligands, including long chain fatty acid (LCFA), advanced glycation end products (AGE), thrombospondin- 1 and thrombospondin-5, oxidized low-density lipoproteins (oxLDLs), high density lipoprotein (HDL), phosphatidylserine, apoptotic cells, P-amyloid fibrils (fA0), collagens I and IV, serum amyloid A (SAA), hexarelin, and Plasmodium falciparum- infected erythrocytes, among others. CD36 is required for the anti-angiogenic effects of thrombospondin- 1 in the corneal neovascularization assay. It plays a role in lipid metabolism and has been identified as a fatty acid translocase necessary for the binding and transport of long- chain fatty acids (LCFA) in cells and tissues. Upon ligand binding, CD36 transduces signals that mediate a wide range of proinflammatory cellular responses. CD36 plays a significant role in the initiation and pathogenesis of chronic inflammatory diseases such as Alzheimer’s disease and atherosclerosis. In fact, several studies suggests that CD36 serves as a signalling hub for lipid homeostasis (Zhou et al. (2008) Gastroenterology 134: 556-67; Kim and Dyck (2016) Biochim. Biophys. Acta 1860: 1450-60), immunological responses (Kumar et al. (2012) Parasite Immunol. 34: 372-82; Urban et al. (2001) Proc. Natl. Acad. Sci. USA 98: 8750-5), and programming of energy availability (Samovski et al. (2015) Diabetes 64: 353-9; Nagendran et al. (2013) J. Mol. Cell. Cardiol. 63: 180-8). The human CD36 gene encodes a single-chain 472 amino acid residue protein containing both an N- and a C-terminal cytoplasmic tail and an extracellular loop. CD36 specificity of action in different tissues likely involves its association with co-receptors, and therefore is strongly dependent on cellular microenvironment. Moreover, CD36 is overexpressed particularly in pro-inflammatory conditions.

[0197] Under steady state conditions, CD36 is not expressed or is present at low density in the cell surface, in which case C4BP(0-) and PRP6-HO7 would not affect CD36 function. Nevertheless, in pro -inflammatory conditions, CD36 would be overexpressed, and its cell surface density would increase to the point to allow C4BP(0-) or PRP6-HO7 intervention through CD36 clustering.

[0198] Method for determining the expression of CD36 on the surface of monocytes and / or macrophages are widely known in the art, for example by flow cytometry as shown in Example 4. Alternatively, other methods can be used for determining the presence of CD36 on the surface of monocytes or macrophages such as immunohistochemistry or Western blot analysis of isolated cell membranes.

[0199] “CD36 expression”, as used herein means that the level of CD36 on the surface of monocytes or macrophages is detectable by any of the above referred detection methods. In a preferred embodiment, it is considered that a subject has CD36 expression wherein the level of CD36 is at least 2%, at least 5%, at least 10%, at least 15 %, at least 20%, at least 25 %, at least 30 %, at least 35 % or at least 40 % more than the level of CD36 expression on the surface of monocytes or macrophages of a subject having CD36 deficiency type I. CD36 deficiency type I as used herein means CD36 is not expressed in either platelets, monocytes, macrophages or other tissue cells. In a preferred embodiment CD36 deficiency type I means that CD36 is not expressed on monocytes or macrophages, preferably means that the expression of CD36 in monocytes or macrophages is zero.

[0200] In a preferred embodiment, the disease involving an undesired activation of the immune system is selected from the group consisting of an immunoinflammatory disease, sepsis, transplant rejection, graft-versus-host disease a hypersensitivity disease, autoimmune and autoinflammatory diseases, liver diseases, metabolic diseases, cardiovascular diseases, cancer, metastatic cancer, renal diseases, infectious diseases, central nervous system diseases, bone diseases, eye diseases, fetal / neonatal alloimmune thrombocytopenia (FNAIT), skin scarring, and oxidized phospholipid-driven lung injury.

[0201] The term “immunoinflammatory disease”, as used herein, refers to inflammatory diseases and disorders in which immune cells and / or cytokines are involved in the pathophysiology of the disease or disorder. Examples of immunoinflammatory diseases include conditions such as rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, acute respiratory distress syndrome and asthma. The term immunoinflammatory disease includes both acute and chronic inflammatory disorders. The term "acute inflammatory disorder" is intended to include disorders and episodes of disorders, characterized by rapid onset of symptoms associated with an inflammatory response and relatively short duration of symptoms, whereas a "chronic inflammatory disorder" is intended to include disorders characterized by the continued presence of symptoms associated with an inflammatory response and ongoing duration of symptoms. Immuno inflammatory diseases which can be treated with the methods according to the present invention include, without limitation, cardiovascular diseases such as infarct or stroke, atherosclerosis, pulmonary fibrosis, rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, acute respiratory distress syndrome, asthma, and cancer. Also comprised within the immunoinflammatory diseases that can be treated according to the present invention are diseases which appear during pregnancy such as pre-eclampsia and eclampsia. Pre-eclampsia is a pregnancy-related disease characterised by hypertension, proteinuria and oedema. Pre-eclampsia is understood and shall be defined herein to encompass and reside within a spectrum of pre-eclampsia disorders, including placental insufficiency, intrauterine growth retardation, early miscarriage, preterm birth, intrauterine death and eclampsia. In a preferred embodiment, the autoinflammatory disease is inflammatory bowel disease.

[0202] The term “sepsis”, as used herein, refers to a systemic host response to microorganisms in previously sterile tissues characterized by end-organ dysfunction away from the primary site of infection. To qualify as sepsis, there must be an infection suspected or proven (by culture, stain, or polymerase chain reaction (PCR)), or a clinical syndrome pathognomonic for infection. Specific evidence for infection includes WBCs in normally sterile fluid (such as urine or cerebrospinal fluid (CSF), evidence of a perforated viscus (free air on abdominal x-ray or CT scan, signs of acute peritonitis), abnormal chest x-ray (CXR) consistent with pneumonia (with focal opacification), or petechiae, purpura, or purpura fulminans. The more critical subsets of sepsis are severe sepsis (sepsis with acute organ dysfunction) and septic shock (sepsis with refractory arterial hypotension). As an alternative, when two or more of the systemic inflammatory response syndrome criteria are met without evidence of infection, patients may be diagnosed simply with "SIRS." Patients with SIRS and acute organ dysfunction may be termed "severe SIRS." Patients are defined as having "severe sepsis" if they have sepsis plus signs of systemic hypoperfusion: either end-organ dysfunction or serum lactate greater than 4 mmol / dL. Other signs include oliguria and altered mental status. Patients are defined as having septic shock if they have sepsis plus hypotension after aggressive fluid resuscitation (typically upwards of 6 liters or 40 ml / kg of crystalloid). Examples of end-organ dysfunction include acute lung injury or acute respiratory distress syndrome, encephalopathy, or dysfunction affecting liver (disruption of protein synthetic function and metabolic functions), kidney (oliguria and anuria, electrolyte abnormalities, volume overload), and heart (systolic and diastolic heart failure).

[0203] Suitable sepsis conditions that can be treated with the compounds according to the present invention include, without limitation, severe sepsis and septic shock. In one embodiment, the condition associated with sepsis syndrome is selected from the group consisting of an organ dysfunction, preferably a kidney dysfunction or a liver dysfunction, a multiple organ dysfunction syndrome (MODS), an acute respiratory distress syndrome (ARDS), and disseminated intravascular coagulation (DIC).

[0204] Sepsis may be induced by a bacterium or more than one bacterium selected from the group consisting of Gram-negative bacteria and Gram-positive bacteria. Preferably, the Gram-negative bacterium is selected from the group consisting of Escherichia coli, Klebsiella species, Serratia species, Enterobacter species, Proteus species, Pseudomonas aeruginosa, Haemophilus influenzae, Neisseria species, and Listeria species. Alternatively, the Gram-positive bacterium is selected from the group consisting of Staphylococcus aureus, Streptococcus pneumoniae, coagulase-negative Staphylococci, Enterococcus species, Streptococcus pyogenes, and Streptococcus viridans. In one embodiment, the sepsis syndrome is induced by LPS. In yet another embodiment, the sepsis is induced by a microorganism or more than one microorganism selected from the group consisting of anaerobic bacteria, fungi, rickettsiae, chlamydiae, mycoplasma, spirochetes, and viruses.

[0205] In a preferred embodiment, the immunological disease is an autoimmune disease.

[0206] The term "autoimmune disease", "disease associated with immune dysfunction / dysregulation" or "immune inflammatory disease" is used throughout the specification to refer to a pathogenic condition in which the patients’ immune system results in disease from a self antigen (autoimmunity) or a foreign antigen (immune dysfunction / dysregulation or immune inflammatory disease). Autoimmunity is present in everyone to some extent. It is usually harmless and probably a universal phenomenon of vertebrate life. However, autoimmunity can be the cause of a broad spectrum of human illnesses, known as autoimmune diseases. This concept of autoimmunity as the cause of human illness is relatively new, and it was not accepted into the mainstream of medical thinking until the 1950s and 1960s. Autoimmune diseases are, thus, defined when the progression from benign autoimmunity to pathogenic autoimmunity occurs. This progression is determined by both genetic influences and environmental triggers. The concept of autoimmunity as the actual cause of human illness (rather than a consequence or harmless accompaniment) can be used to establish criteria that define a disease as an autoimmune disease. Autoimmune diseases or diseases which are characterized as involving immune dysfunction or dysregulation (immune inflammatory disease), which may be treated by the present invention include systemic lupus erythematosus (SLE), lupus nephritis, central nervous system (CNS) lupus, diabetes mellitus (type I), asthma, ulcerative colitis, Crohn’s disease, Grave's disease, arthritis, including rheumatoid arthritis and osteoarthritis, pernicious anemia, inflammatory bowel disease and multiple sclerosis, among numerous others. Numerous autoimmune diseases may be treated using the method of the present invention including autoimmune blood diseases, including pernicious anemia, autoimmune hemolytic anemia, aplastic anemia, idiopathic thrombocytopenic purpura, ankylosing spondilitis; autoimmune diseases of the musculature including polymyositis and dermatomyositis; autoimmune diseases of the ear including autoimmune hearing loss and Meniere's syndrome; autoimmune eye diseases, including Mooren's disease, Reiter's syndrome and Vogt-Koyanagi-Harada disease; autoimmune diseases of the kidney including glomerulonephritis, IgA nephropathy, and lupus nephritis; diabetes mellitus (type I); autoimmune skin diseases including pemphigus (autoimmune bullous diseases), such as pemphigus vulgaris, pemphigus foliaceus, pemphigus erythematosus, bullous pemphigoid, vitiligo, epidermolysis bullosa acquisita, psoriasis and alopecia areata; cardiovascular autoimmune diseases, including autoimmune myocarditis, vasculitis including Churg-Strauss syndrome, giant cells arteritis, Kawasaki's disease, polyarteritis nodosa, Takayasu's arteritis and Wegener's granulomatosis; endocrine autoimmune diseases, including Addison's disease, autoimmune hypoparathyroidism, autoimmune hypophysitis, autoimmune oophoritis, autoimmune orchitis, Grave's Disease, Hashimoto's thyroiditis, polyglandular autoimmune syndrome type 1 (PAS-1), polyglandular autoimmune syndrome type 2 (PAS-2), and polyglandular autoimmune syndrome type 3 (PAS-3); autoimmune gastroenteric diseases including autoimmune hepatitis, primary biliary cirrhosis, inflammatory bowel disease, celiac disease, Crohn's disease; autoimmune nervous diseases, including multiple sclerosis, myasthenia gravis, Guillan-Barre syndrome and chronic inflammatory demyelinating neuropathy; and systemic autoimmune diseases including systemic lupus erythematosus, antiphospholid syndrome, autoimmune lymphoproliferative disease, autoimmune polyendocrinopathy, Bechet's disease, Goodpasture's disease, arthritis, including rheumatoid arthritis, osteoarthritis and septic arthritis, sarcoidosis, scleroderma and Sjogren's syndrome and psoriasis among others.

[0207] In an embodiment, the autoimmune disease is lupus erythematosus. The expression “lupus erythematosus”, as used herein, refers to a name given to a collection of autoimmune diseases that have common symptoms that affect joints, skin, kidneys, blood cells, heart and lungs. Lupus erythematosus may manifest as systemic disease or in a purely cutaneous form also known as incomplete lupus erythematosus. Lupus has four main types: systemic, discoid, drug-induced and neonatal. The term “lupus erythematosus” in the context of the present invention encompasses, without limitation, acute cutaneous lupus erythematosus, subacute cutaneous lupus erythematosus, discoid lupus erythematosus (chronic cutaneous), childhood discoid lupus erythematosus, generalized discoid lupus erythematosus, localized discoid lupus erythematosus, chilblain lupus erythematosus (Hutchinson), lupus erythematosus-lichen planus overlap syndrome, lupus erythematosus panniculitis (lupus erythematosus profundus), tumid lupus erythematosus, verrucous lupus erythematosus (hypertrophic lupus erythematosus), cutaneous lupus mucinosis, complement deficiency syndromes, drug- induced lupus erythematosus, neonatal lupus erythematosus and systemic lupus erythematosus. The most common severe form is systemic lupus erythematosus.

[0208] In a preferred embodiment, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus, lupus nephritis, rheumatoid arthritis, inflammatory bowel disease and ulcerative colitis; more preferably is selected from the group consisting of systemic lupus erythematosus, lupus nephritis, inflammatory bowel disease and rheumatoid arthritis; more preferably from systemic lupus erythematosus, lupus nephritis and rheumatoid arthritis. In a preferred embodiment, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus and lupus nephritis.

[0209] In a preferred embodiment the autoimmune disease is systemic lupus erythematosus. The expression “systemic lupus erythematosus” or “SLE”, as used herein, refers to a systemic autoimmune disease in which the body’s immune system mistakenly attacks healthy tissue in many parts of the body. Symptoms vary from person to person and may be mild to severe. Common symptoms include painful and swollen joints, fever, chest pain, hair loss, mouth ulcers, swollen lymph nodes, feeling tired, and a red rash which is most commonly on the face. Often there are periods of illness, called flares, and periods of remission when there are few symptoms. Almost everyone with SLE has joint pain and swelling. Some develop arthritis. SLE often affects the joints of the fingers, hands, wrists, and knees. Renal disease in SLE carries significant morbidity and mortality. Acute or chronic renal impairment may develop with lupus nephritis, leading to acute or end-stage kidney failure.

[0210] In a preferred embodiment the autoimmune disease is lupus nephritis.

[0211] The expression “lupus nephritis” or “LN”, also known as SLE nephritis, is an inflammation of the kidneys caused by systemic lupus erythematosus. It is a type of glomerulonephritis in which the glomeruli become inflamed. As the result of SLE, the cause of glomerulonephritis is said to be secondary and has a different pattern and outcome from conditions with a primary cause originating in the kidney. General symptoms of lupus nephritis include fever, oedema, high blood pressure, joint pain, muscle pain, malar rash and foamy urine.

[0212] In another preferred embodiment, the autoimmune disease is inflammatory bowel disease, more preferably ulcerative colitis.

[0213] The expression “inflammatory bowel disease”, as used herein, refers to a group of inflammatory conditions of the colon and small intestine. Crohn’s disease and ulcerative colitis are the principal types of inflammatory bowel disease. Crohn’s disease affects the small intestine and large intestine, as well as the mouth, esophagus, stomach and the anus; whereas ulcerative colitis primarily affects the colon and the rectum. The symptoms can be: abdominal pain, diarrhea, rectal bleeding, severe internal cramps / muscle spasms in the region of the pelvis and weight loss.

[0214] In another preferred embodiment, the autoimmune disease is rheumatoid arthritis.

[0215] The expression “rheumatoid arthritis” or “RA”, as used herein, refers to a longterm systemic autoimmune disorder characterized by chronic inflammation of the joints and the subsequent destruction of cartilage and bone. It typically results in warm, swollen, and painful joints. Pain and stiffness often worsen following rest. Most commonly, the wrist and hands are involved, with the same joints typically involved on both sides of the body. The disease may also affect other parts of the body. This may result in a low red blood cell count, inflammation around the lungs, and inflammation around the heart. Fever and low energy may also be present. Often, symptoms come on gradually over weeks to months. RA primarily starts as a state of persistent cellular activation leading to autoimmunity and immune complexes in both joints and other organs where it manifests. The initial site of disease is the synovial membrane, where swelling and congestion leads to infiltration by immune cells. The various phases of progression of RA are:

[0216] • Initiation phase, due to non-specific inflammation.

[0217] • Amplification phase, due to T cell activation

[0218] • Chronic inflammatory phase with tissue injury, due to cytokines IL- 1, TNF-alpha and IL-6.

[0219] The expression “transplant rejection”, as used herein, refers to an immune condition in which a transplanted cell, tissue, or organ is not accepted by the body of the transplant recipient. The expression transplant rejection encompasses both acute and chronic transplant rejection.

[0220] "Acute rejection or AR" is the rejection by the immune system of a tissue transplant recipient when the transplanted tissue is immunologically foreign. Acute rejection is characterized by infiltration of the transplanted tissue by immune cells of the recipient, which carry out their effector function and destroy the transplanted tissue. The onset of acute rejection is rapid and generally occurs in humans within a few weeks after transplant surgery.

[0221] "Chronic transplant rejection or CR" generally occurs in humans within several months to years after engraftment, even in the presence of successful immunosuppression of acute rejection. Fibrosis is a common factor in chronic rejection of all types of organ transplants. Chronic rejection can typically be described by a range of specific disorders that are characteristic of the particular organ. For example, in lung transplants, such disorders include fibroproliferative destruction of the airway (bronchiolitis obliterans); in heart transplants or transplants of cardiac tissue, such as valve replacements, such disorders include fibrotic atherosclerosis; in kidney transplants, such disorders include, obstructive nephropathy, nephrosclerosis, tubulointerstitial nephropathy; and in liver transplants, such disorders include disappearing bile duct syndrome. Chronic rejection can also be characterized by ischemic insult, denervation of the transplanted tissue, hyperlipidemia and hypertension associated with immunosuppressive drugs.

[0222] As is known in the transplantation field, the transplant organ, tissue or cell(s) may be allogeneic or xenogeneic, such that the grafts may be allografts or xenografts. A feature of the graft tolerant phenotype detected or identified by the subject methods is that it is a phenotype which occurs without immunosuppressive therapy, i.e., it is present in a host that is not undergoing immunosuppressive therapy such that immunosuppressive agents are not being administered to the host. The transplant graft maybe any solid organ and skin transplant. Examples of organ transplants that can be treated by the methods described herein include but are not limited to kidney transplant, pancreas transplant, liver transplant, heart transplant, lung transplant, intestine transplant, pancreas after kidney transplant, and simultaneous pancreas-kidney transplant.

[0223] The methods according to the present invention are also suitable for the prevention and / or treatment of delayed Graft Function (DGF) due to ischemiareperfusion injury. The term “delayed graft function”, as used herein, refers to a form of acute renal failure resulting in post- transplantation oliguria, increased allograft immunogenicity and risk of acute rejection episodes, and decreased long-term survival. DGF may be caused by different factors related to the donor and prerenal, renal, or postrenal transplant factors related to the recipient. However, a major cause of delayed graft function is ischaemia and reinstitution of blood flow in ischaemically damaged kidneys after hypothermic preservation.

[0224] The term “graft-versus-host disease” or GVHD, as used herein, refers to a condition that occurs when T cells present in donor tissue attack the host, or recipient, of the grafted cells or tissue. Any type of GVHD can be treated by the therapeutic agents of the present invention, including acute GVHD and chronic GVHD.

[0225] The term "hypersensitivity disease" refers to a condition in which the subject has an abnormal sensitivity to an innocuous agent, known as allergen. Hypersensivity disease can be categorized into four types, Type I, Type II, Type III and Type IV. Type I is described as atopic or anaphylactic which results from a release of mediators from IgE-sensitized basophils and mast cells. Type II is described as cytotoxic which involves complement-fixing antibody with cell lysis or antibody-dependent cellular cytotoxocity. Type III is described as immune-complex-mediated which is associated with soluble antigen-antibody complexes. Type IV is described as cell-mediated or delayed hypersensitivity which results from a release of lymphokines by sensitized T lymphocytes after contact with an antigen.

[0226] In a preferred embodiment, the immunological disease is inflammatory bowel disease, more preferably ulcerative colitis. In another embodiment, the immunological disease is Crohn’s disease.

[0227] In a preferred embodiment, the disease involving an undesired activation of the immune system is selected from the group consisting of autoimmune and autoinflammatory diseases, liver diseases, metabolic diseases, cardiovascular diseases, cancer, metastasic cancer, renal diseases, infectious diseases, central nervous system diseases, bone diseases, eye diseases, fetal / neonatal alloimmune thrombocytopenia (FNAIT), skin scarring, and oxidized phospholipid-driven lung injury.

[0228] In a more preferred embodiment, the metabolic disease is selected from the group consisting of hepatic steatosis, metabolic syndrome, obesity, diabetes, hyperlipidemia, atherosclerosis, nephropathy, retinopathy, peripheral neuropathy, nonalcoholic fatty liver disease (NAFLD); wherein the cardiovascular disease is selected from the group consisting of stroke, heart failure, vasculitis, venous thromboembolism and diabetic cardiomyopathy; wherein the cancer is selected from the group consisting of radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma and lung cancer; wherein the renal disease is selected from the group consisting of chronic kidney disease and lithogenic-induced gallstones; wherein the infectious disease is selected from the group consisting of COVID-19, hepatitis B and C, HIV-induced AIDS and malaria; wherein the central nervous system disease is Alzheimer’s disease; wherein the bone disease is selected from the group consisting of osteoporosis and periodontitis; wherein the eye disease is selected from the group consisting of age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation and retinal inflammation / degeneration. In a more preferred embodiment, the disease is selected from the group consisting of autoimmune and autoinflammatory diseases, hepatic steatosis and cancer.

[0229] In another embodiment, the compound is administered in combination with one or more therapeutic agents useful in the treatment of a disease involving an undesired activation of the immune system, preferably said therapeutic agent is selected from the group consisting of cyclosporine A, tacrolimus, methotrexate, thiopurines, anti-TNF agents, infliximab, adalimumab, certolizumab, golimumab, etanercept, rituximab, epratuzumab, belimumab, rapamycin, anti-interferon antibodies, tocilizumab, laquinimod, tabalumab, ofatumumab, ixekizumab, brodalumab, briakinumab, sarilumab, rilonacept, anifrolumab, cyclophosphamide, mycophenolate mofetil, azathioprine, anticalcineurinics, prednisolone, methylprednisolone, vitamin D, vasoactive intestinal peptide, hydroxychloroquine, chloroquine, ocrelizumab, atacicept, abatacept, alemtuzumab, sirukumab, eculizumab and T cell vaccine.

[0230] In another aspect, the disease to be prevented or treating with the compounds of the invention is selected the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury.

[0231] In a preferred embodiment, the diseases to be prevented or treated with the compounds of the invention are diseases related with an increase expression in CD36. “Increase expression”, as used herein relates to the level of CD36 higher than 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, 100%, 200%, 300% or more, in comparison to the level of CD36 in a control sample.

[0232] Methods for detecting the expression of a gene, in the case CD36, can be based on detecting mRNA or protein, or they also can be based on determining the mRNA levels or protein levels and the levels of variants thereof, in a sample as a whole, in cells of a sample and / or in the non-cellular fraction of a sample.

[0233] Methods for detecting mRNA are well known in the art and include without limitation, standard assays for determining mRNA expression levels such as qPCR, RT- PCR, RNA protection analysis, Northern blot, RNA dot blot, in situ hybridization, microarray technology, tag based methods such as serial analysis of gene expression (SAGE) including variants such as LongSAGE and SuperSAGE, microarrays, fluorescence in situ hybridization (FISH), including variants such as Flow-FISH, qFiSH and double fusion FISH (D-FISH), and the like. Preferably quantitative or semi- quantitative RT-PCR is preferred. Real-time quantitative or semi-quantitative RT-PCR is particularly advantageous.

[0234] In case the mRNA is measured in a biological sample, said biological sample may be treated to physically, mechanically or chemically disrupt tissue or cell structure, to release intracellular components into an aqueous or organic solution to prepare nucleic acids for further analysis. The nucleic acids are extracted from the sample (e.g., cell or tissue prepared from the subject) by procedures known to the skilled person and commercially available, for example using lytic enzymes or chemical solutions or extracted by nucleic-acid-binding resins following the manufacturer's instructions. RNA is then extracted from frozen or fresh samples by any of the methods typical in the art, for example, Sambrook, J., et al., 2001. Molecular cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, N.Y., Vol. 1-3. Preferably, care is taken to avoid degradation of the RNA during the extraction process.

[0235] The expression level can be determined using mRNA obtained from a formalin- fixed, paraffin-embedded tissue sample. mRNA may be isolated from an archival pathological sample or biopsy sample which is first deparaffinized. An exemplary deparaffinization method involves washing the paraffinized sample with an organic solvent, such as xylene. Deparaffinized samples can be rehydrated with an aqueous solution of a lower alcohol. Suitable lower alcohols, for example, include methanol, ethanol, propanols and butanols. Deparaffinized samples may be rehydrated with successive washes with lower alcoholic solutions of decreasing concentration, for example. Alternatively, the sample is simultaneously deparaffinized and rehydrated. The sample is then lysed and RNA is extracted from the sample. Samples can be also obtained from fresh tumor tissue such as a resected tumor. In a particular embodiment, samples can be obtained from fresh tumor tissue or from OCT embedded frozen tissue.

[0236] In order to normalize the values of mRNA expression among the different samples, it is possible to compare the expression levels of the mRNA of interest in the test samples with the expression of a control RNA. A “control RNA”, as used herein, relates to RNA whose expression levels do not change or change only in limited amounts. Preferably, the control RNA is mRNA derived from housekeeping genes and which code for proteins which are constitutively expressed and carry out essential cellular functions. Preferred housekeeping genes for use in the present invention include 18-S ribosomal protein, P-2-microglobulin, ubiquitin, cyclophilin, GAPDH, PSMB4, tubulin and P-actin.

[0237] The relative gene expression quantification may be calculated according to the comparative threshold cycle (Ct) method using a housekeeping gene as an endogenous control and commercial RNA controls as calibrators. Final results are determined according to the formula 2-(ACt sample-ACt calibrator), where ACt values of the calibrator and sample are determined by subtracting the Ct value of the target gene from the value of the control gene.

[0238] Alternatively, it is also possible to determine the expression level of CD36 by means of the determination of the expression levels of the proteins encoded by said gene, since if the expression of the gene is increased, an increase of the amount of corresponding proteins should occur and if the expression of the gene is decreased, a decrease of the amount of corresponding proteins should occur.

[0239] Virtually any conventional method can be used within the frame of the invention to detect and quantify the levels of proteins. By way of a non-limiting illustration, the expression levels are determined by means of antibodies with the capacity for binding specifically to the protein to be determined (or to fragments thereof containing the antigenic determinants) and subsequent quantification of the resulting antigen-antibody complexes. The antibodies that are going to be used in this type of assay can be, for example, polyclonal sera, hybridoma supernatants or monoclonal antibodies, antibody fragments, Fv, Fab, Fab’ and F(ab’)2, scFv, diabodies, triabodies, tetrabodies and humanized antibodies. At the same time, the antibodies may or may not be labeled. Illustrative, but non-exclusive, examples of markers that can be used include radioactive isotopes, enzymes, fluorophores, chemoluminescent reagents, enzyme cofactors or substrates, enzyme inhibitors, particles, dyes, etc. There is a wide variety of well-known assays that can be used in the present invention, using non-labeled antibodies (primary antibody), labeled antibodies (secondary antibodies); these techniques include Westernblot or immunoblot, ELISA (enzyme-linked immunosorbent assay), RIA (radioimmunoassay), competitive EIA (enzyme immunoassay), DAS-ELISA (double antibody sandwich ELISA), immunocytochemical and immunohistochemical techniques, immunofluorescence, techniques based on the use of biochips or protein microarrays including specific antibodies or assays based on the colloidal precipitation in formats such as reagent strips. Other forms of detecting and quantifying the proteins include affinity chromatography techniques, ligand-binding assays, etc.

[0240] On the other hand, the determination of the levels of a protein can be carried out by constructing a tissue microarray (TMA) containing the subject samples assembled, and determining the expression levels of the corresponding protein by immunohistochemistry techniques. Immuno staining intensity can be evaluated by two or more different pathologists and scored using uniform and clear cut-off criteria, in order to maintain the reproducibility of the method. Discrepancies can be resolved by simultaneous re-evaluation. Briefly, the result of immuno staining can be recorded as negative expression (0) versus positive expression, and low expression (1+) versus moderate (2+) and high (3+) expression, taking into account the expression in tumor cells and the specific cut-off for each marker. As a general criterion, the cut-offs are selected in order to facilitate reproducibility, and when possible, to translate biological events. Alternatively, the immuno staining intensity can be evaluated by using imaging techniques and automated methods such as those disclosed in Rojo, M.G. et al. (Folia Histochem. CytobioL 2009; 47: 349-54) or Mulrane, L. et al. (Expert Rev. Mol. Diagn. 2008; 8: 707-25). Disease related with increased levels of CD36 is for example cancer and particularly myeloid leukemia, monocytic leukemia, ovarian cancer (Ladanyi et al. (2018) Oncogene 37: 2285-301), breast cancer (Gyamfi et al. (2021) Breast Cancer 7: 129), radiotherapy and / or chemotherapy resistant cancer, cancer resistance to HER2- targeted therapies (Feng et al. (2019) Cell Reports 29: 3405-20), gastric cancer metastasis (Jiang et al. (2019) Theranostics 9: 5359-73; Wang et al. (2019) EBioMedicine 45: 108-123), oral squamous cell carcinoma (OSCC) (Pascual et al. (2021) Nature 599: 485-90), colorectal cancer metastasis (Drury et al. (2022) Cancers 14: 252), melanoma and breast cancer-derived tumors (Pascual et al. (2017) Nature 541: 41-5), colon adenocarcinoma, metastatic cancer, and lung cancer ((Wang et al. (2020) Nat Immunol. 21: 298-308).

[0241] Metabolic diseases are also related to increased levels of CD36, for example metabolic syndrome, obesity, diabetes, insulin resistance, atherosclerosis, nephropathy, retinopathy, peripheral neuropathy, diabetic cardiomyopathy; non-alcoholic fatty liver disease (NAFLD)). (Karunakaran et al. (2021) Cells 10: 1833; Navas-Madronal et al. (2020) Int. J. Mol. Sci. 21: 7360; Puchalowicz and Rac (2020) Cells 9: 1877; Wilson et al. (2016) Endocrinology 157: 570-85; Wang et al. (2014) PLOS One 9: el03071; Rada et al. (2020) Cell Death Dis. (2020) 11: 802); Angin et al. (2012) Biochem. J. 448:43- 53; Yang et al. (2018) J. Biol. Chem. 293: 13338-48). Other diseases related with increased levels of CD36 are for example lithogenic-induced gallstones (Xie et al. (2017) J. Lipid Res. 58: 1692-701), COVID-19 (Gomes Dias et al. (2020) PLoS Pathog. 16: e!009127), hepatitis B and C (Cheng et al. (2016) Sci. Rep. 6: 21808) and (Huang et al. (2016) PLoS One 11: e0164787), HIV-induced AIDS (Berre et al. (2013) J. Exp. Med. 210: 2523-38) and malaria (Bachmann et al. (2022) Microorganisms 10: 2356). Also diseases such as Alzheimer's disease (Dobri et al. (2021) Neuroscience 453: 301- 11), osteoporosis and periodontitis ((Koduru et al. (2018) J. Biol. Chem 293: 15055-69; Lu et al. (2017) Oral Dis. 23: 210-18; Rekhi et al. (2021) 126: 105129); age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation and retinal inflammation / degeneration ((Lavalette et al. (2020) Front. Immunol. 10: 3032) and (Yang et al. (2023) Cells 12: 171); skin scarring (Griffin et al. (2021) Sci. Transl. Med. 13: eabb3312), fetal / neonatal alloimmune thrombocytopenia (FNAIT) ((Xu et al. (2021) Blood 138: 1757-67); and oxidized phospholipid-driven lung injury.

[0242] In a more preferred embodiment, the disease is selected from the group consisting of autoimmune and auto inflammatory diseases, hepatic steatosis and cancer, more preferably hepatic steatosis.

[0243] “Hepatic steatosis”, as used herein relates to a condition marked by an abnormal buildup of triglycerides in the liver, is the precursor to NAFLD. It is generally agreed that CD36 contributes significantly to hepatic steatosis by taking part in fatty acid uptake as well as triglyceride storage and secretion.

[0244] The invention also relates to the following aspects:

[0245] [1]. An oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

[0246] [2], The oligomer for use according to aspect 1, wherein the oligomer comprises at least six polypeptides.

[0247] [3]. The oligomer for use according to any one of aspects 1 or 2 wherein the oligomer is an homooligomer.

[0248] [4], The oligomer for use according to any one of aspects 1 to 3, wherein said polypeptide does not comprise one or more of the domains selected from the group consisting of CCP1, CCP2, CCP3, CCP4, CCP5, CCP7 and CCP8 of the C4BP alpha chain.

[0249] [5]. The oligomer for use according to any one of aspects 1 to 4, wherein the polypeptide comprises a full-length C4BP alpha chain.

[0250] [6]. The oligomer for use according to any one of aspects 1 to 5, wherein the CCP6 domain of the C4BP alpha chain is SEQ ID NO: 1.

[0251] [7]. The oligomer for use according to any one of aspects 1 to 6, wherein the functionally equivalent variant of the CCP6 domain of the C4BP alpha chain is a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO: 2, 3, 4 and 5.

[0252] [8]. The oligomer for use according to any one of aspects 1 to 7, wherein the oligomerization domain is SEQ ID NO: 6 or a functionally equivalent variant thereof, preferably SEQ ID NO: 6.

[0253] [9]. The oligomer for use according to any one of aspects 1 to 8, wherein the polypeptide consists of SEQ ID NO: 7 or a functionally equivalent variant thereof, preferably SEQ ID NO: 7.

[0254]

[0010] . The oligomer for use according to any one of aspects 1 to 9, wherein the polypeptide consists of a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9 and a functionally equivalent variant thereof.

[0255]

[0011] . The oligomer for use according to any one of aspects 1 to 9, wherein the polypeptide consists of SEQ ID NO: 10 or a functionally equivalent variant thereof, preferably SEQ ID NO: 10.

[0256]

[0012] , The oligomer for use according to any one of aspects 1, 3,5 or 6, wherein the polypeptide consists of SEQ ID NO: 11 or a functionally equivalent variant thereof.

[0257]

[0013] . The oligomer for use according to any one of aspects 1, 3, 5 or 6, wherein the oligomer is a C4BP isoform selected from the group consisting of a?Po and aePo.

[0258]

[0014] , The oligomer for use according to any one of aspects 1 to 13, wherein the disease involving an undesired activation of the immune system is selected from the group consisting of immunoinflammatory disease, sepsis, autoimmune disease, transplant rejection, graft-versus-host disease, hypersensitivity disease, autoinflammatory diseases, liver diseases, metabolic diseases, cardiovascular diseases, cancer, metastasic cancer, renal diseases, infectious diseases, central nervous system diseases, bone diseases, eye diseases, fetal / neonatal alloimmune thrombocytopenia (FNAIT), skin scarring, and oxidized phospholipid-driven lung injury; preferably selected from the group consisting of immunoinflammatory disease, sepsis, autoimmune disease, transplant rejection, graft-versus-host disease, and hypersensitivity disease.

[0259]

[0015] . The oligomer for use according to aspect 14, wherein the metabolic disease is selected from the group consisting of hepatic steatosis, metabolic syndrome, obesity, diabetes, hyperlipidemia, atherosclerosis, nephropathy, retinopathy, peripheral neuropathy, non-alcoholic fatty liver disease (NAFLD); wherein the cardiovascular disease is selected from the group consisting of stroke, heart failure, diabetic cardiomyopathy, vasculitis and venous thromboembolism; wherein the cancer is selected from the group consisting of radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma and lung cancer; wherein the renal disease is selected from the group consisting of chronic kidney disease and lithogenic-induced gallstones; wherein the infectious disease is selected from the group consisting of COVID- 19, hepatitis B and C, HIV-induced AIDS and malaria; wherein the central nervous system disease is Alzheimer’s disease; wherein the bone disease is selected from the group consisting of osteoporosis and periodontitis; wherein the eye disease is selected from the group consisting of age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation and retinal inflammation / degeneration.

[0260]

[0016] . The oligomer for use according to aspect 14, wherein the disease is selected from the group consisting of autoimmune and autoinflammatory diseases, hepatic steatosis and cancer.

[0261]

[0017] . The oligomer for use according to any one of aspects 1 to 16, wherein the compound is to be administered by subcutaneous route.

[0262]

[0018] . An oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease selected from the group consisting of hepatic steatosis, metastatic cancer, radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma, lung cancer, metabolic syndrome, obesity, retinopathy, diabetic cardiomyopathy, non-alcoholic fatty liver disease (NAFLD), vasculitis, venous thromboembolism, lithogenic-induced gallstones, COVID-19, hepatitis B and C, HIV- induced AIDS, malaria, Alzheimer's disease, osteoporosis, periodontitis, age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation, retinal inflammation / degeneration, skin scarring, fetal / neonatal alloimmune thrombocytopenia (FNAIT) and oxidized phospholipid- driven lung injury.

[0263]

[0019] . The oligomer for use according to aspect 18, wherein the oligomer comprises at least six polypeptides.

[0264]

[0020] . The oligomer for use according to any one of aspects 18 or 19, wherein the oligomer is an homooligomer.

[0265]

[0021] , The oligomer for use according to any one of aspects 18 to 20, wherein said polypeptide does not comprise one or more of the domains selected from the group consisting of CCP1, CCP2, CCP3, CCP4, CCP5, CCP7 and CCP8 of the C4BP alpha chain.

[0266]

[0022] , The oligomer for use according to any one of aspects 18 to 20, wherein the polypeptide comprises a full-length C4BP alpha chain.

[0267]

[0023] . The oligomer for use according to any one of aspects 18 to 22, wherein the CCP6 domain of the C4BP alpha chain is SEQ ID NO: 1.

[0268]

[0024] , The oligomer for use according to any one of aspects 18 to 22, wherein the functionally equivalent variant of the CCP6 domain of the C4BP alpha-chain is a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO: 2, 3, 4 and 5.

[0269]

[0025] . The oligomer for use according to any one of aspects 18 to 23, wherein the oligomerization domain is SEQ ID NO: 6 or a functionally equivalent variant thereof, preferably SEQ ID NO: 6.

[0270]

[0026] . The oligomer for use according to any one of aspects 18 to 25, wherein the polypeptide consists of SEQ ID NO: 7 or a functionally equivalent variant thereof, preferably SEQ ID NO: 7.

[0271]

[0027] . The oligomer for use according to any one of aspects 18 to 25, wherein the polypeptide consists of a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9 and a functionally equivalent variant thereof.

[0272]

[0028] . The oligomer for use according to any one of aspects 18 to 25, wherein the polypeptide consists of SEQ ID NO: 10 or a functionally equivalent variant thereof, preferably SEQ ID NO: 10.

[0029] . The oligomer for use according to any one of aspects 18, 19, 20, 22 or 23, wherein the polypeptide consists of SEQ ID NO: 11 or a functionally equivalent variant thereof

[0273]

[0030] . The oligomer for use according to any one of aspects 18, 19, 20, 22 or 23, wherein the oligomer is a C4BP isoform selected from the group consisting of a?Po and aePo.

[0274]

[0031] . The oligomer for use according to any one of aspects 18 to 30, wherein the oligomer is to be administered by subcutaneous route.

[0275] The invention is described in detail by way of the following examples which are to be considered as merely illustrative and not limitative of the scope of the invention.

[0276] EXAMPLES

[0277] Examples 1-9

[0278] MATERIALS AND METHODS

[0279] Monocyte isolation, culture, and differentiation

[0280] Total blood from healthy donors was acquired in the Blood and Tissue Bank (Barcelona, Spain) and their PBMCs were isolated at less than 16 h after extraction through Ficoll-Paque density gradient centrifugation (Cytiva, Marlborough, MA). For all binding and functional assays, monocytes were purified using colloidal super- paramagnetic microbeads conjugated with monoclonal mouse anti-human CD 14 antibodies (Miltenyi Biotec, Auburn, CA) and counted using Perfect Count microspheres (Cytognos SL, Salamanca, Spain). The purity of CD14+cells was tested by CD14 staining with FITC-conjugated anti-CD14 MoAb (TUK4) (Miltenyi Biotec) and flow cytometry analysis through a Gallios Analyzer (Beckman Coulter, Brea, CA), being in all cases > 90%.

[0281] Monocytes were plated in RPMI 1640 supplemented with 100 mg / ml streptomycin, 100 lU / ml penicillin, 2 mM L-glutamine, and 10% heat- inactivated FBS (all from Thermo Fisher, Waltham, MA) (complete medium) at 37 °C under 5% CO2. Monocyte cultures were supplemented with either GM-CSF (800 lU / ml) plus IL-4 (500 lU / ml) (both from Gentaur, Kampenhout, Belgium) for monocyte to monocyte-derived dendritic cell (Mo-DC) differentiation, or with GM-CSF (650 lU / ml) (Gentaur) to generate monocyte-derived macrophages (Mo-macrophages). Unbiased PRP6-HO7-based receptor identification in human monocytes

[0282] To perform ligand-receptor capture (LRC) (Frei et al. (2013) Nat. Protoc. 8: 1321-36), 300 pg of PRP6-HO7 (Serrano et al. (2022) Front. Immunol. 13: 883743), or glycine (Gly) as negative control (quenching reagent), were coupled to 150 pg of the trifunctional crosslinker TriCEPS v.3.0 (Dualsystems Biotech AG, Schlieren, Switzerland), each in a total volume of 750 pl and in 25 m HEPES buffer, pH 8.2 (2 h at 22 °C, under constant gentle agitation).

[0283] Next, 40 x 106Mo-DC differentiating cells / donor (16 h after the initiation of Mo-DC differentiation) were resuspended in ice cold PBS, pH 6.5. The cells were then mildly oxidized by treatment with 1.5 mM NaIO4 at 4 °C for 15 min in the dark under gentle rotation. After oxidation, the cells were washed, resuspended in PBS with Ca2+ / Mg2+, pH 7.4, plus 2-amino-5-methoxybenzoic acid as catalyst, and incubated with TriCEPS-PRP6-HO7, or TriCEPS-Gly at 4 °C with gentle shaking for 90 min in the dark. After labelling, the cells were resuspended again in PBS with Ca2+ / Mg2+, pH 7.4, including the catalyst. For each treatment arm, cells from the 3 different donors were pooled, distributed in two different tubes, pelleted, snap frozen and stored at -80 °C.

[0284] Cells were lysed according to a standard protocol and target proteins were purified using solid phase chromatography. After stringent washing steps to remove unspecific interactions, proteins were reduced, alkylated, and digested with trypsin. The tryptic peptides were collected for LC-MS / MS.

[0285] The corresponding LRC-TriCEPS peptides were analyzed on a Q-Exactive mass spectrometer (Thermo-Fisher Scientific) fitted with an electrospray ion source. Tryptic peptides were measured in data dependent acquisition mode (TOP 12) in a 90 min gradient using a 50 cm C18 packed column. Each sample was injected in triplicate.

[0286] For data analysis, the Progenesis software was used for raw file alignment and feature detection. The Comet search engine was used for spectra identification and the Trans proteomic pipeline was used for statistical validation of putative identifications and protein inference. Upon protein inference, relative quantification of control and ligand samples was performed based on ion extracted intensity and differential protein abundance was tested using a statistical ANOVA model followed by multiple testing correction. This model assumes that the measurement error follows Gaussian distribution and views individual features as replicates of a protein's abundance and explicitly accounts for this redundancy. It tests each protein for differential abundance in all pairwise comparisons of ligand and control samples and reports the p-values. Next, p-values are adjusted for multiple comparisons to control the experiment-wide false discovery rate (FDR). The adjusted p- value (qvalue) obtained for every protein is plotted against the magnitude of the fold enrichment between the two experimental conditions. In the present study, the human proteome database from Uniprot was used for analysis. Results were filtered for membrane associated proteins. This filter consists of all proteins reported to be associated to the membrane and / or secreted.

[0287] Surface plasmon resonance (SPR) spectroscopy

[0288] SPR assays were carried out on a Biacore T200 (Cytiva) with active temperature control at 25 °C following the manufacturer’s protocols. Briefly, the inventors used a protein A sensor chip (Cytiva). The running buffer was PBS plus 0.5% Surfactant P20 (PBS-P Buffer). Human IgG (Sigma-Aldrich, Saint Louis, MO) was diluted in running buffer (2.5 pg / ml) and injected at a flow rate of 5 pl / min for 360 s of contact time. Recombinant Human CD36 / SR B3 Fc Chimera (R&D Systems, Minneapolis, MN) was diluted at 10 pg / ml in running buffer and captured in another flow cell at a flow rate of 5 pl / min for 1040 s. The analyte PRP6-HO7 was diluted in running buffer (final concentrations ranging from 15.62 to 1,000 nM) and flowed across both immobilized human IgG and recombinant Human CD36 / SR B3 Fc Chimera for 90 s at a flow rate of 30 pl / min (association). The chip surface was regenerated by injecting 10 mM Glycine- HC1, pH 1.5, at a flow rate of 10 pl / min for 30 s. Data were analyzed using Biacore T200 evaluation software and fitted using GraphPad Prism software (San Diego, CA).

[0289] PRP6-HO7 cell binding assay

[0290] Monocytes were plated at 2.5 x 105cells / 500 pl in 24-well culture plates (Jet Biofil, Guangzhou, China) with complete RPMI medium supplemented with GM-CSF (800 lU / ml) and IL-4 (500 lU / ml). After 24 h, differentiating Mo-DCs were harvested and 1 x 105cells in 100 pl of basal RPMI (without FBS) were incubated at room temp. (RT) with anti-human CD36 monoclonal antibody (FA6-152) or with mouse monoclonal IgGl kappa isotype control antibody (both from Stemcell Technologies, Vancouver, Canada) at 20 pg / ml for 30 min on a rotator. Next, PRP6-HO7 was added at 10 pg / ml and further incubated at 4 °C for 60 min. After washing with FACS buffer (PBS containing 1% BSA and 0.1% sodium azide), PRP6-HO7 binding to differentiating Mo-DCs was detected by adding 0.5 pl FITC-conjugated anti-Strep-Tag II antibody (GenScript, Piscataway, NJ). Staining with 0.3 pl LIVE / DEAD™ fixable Near-IR Dead Cell dye (Invitrogen, Thermo Fisher) was employed to assess cell viability, which was stopped after 20 min by cell resuspension in 200 pl fixation buffer (PBS + 4% paraformaldehyde). The inventors gated the cells according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Stained and fixed cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ). Subsequent analyses used FlowJo software (Flowjo LLC, Ashland, OR).

[0291] PRP6-HO7 saturation curve

[0292] Mo-DCs at 24 h from the beginning of differentiation were collected at 2 x 105cells / 100 pl in PBS with Ca2+and Mg2+and incubated with increasing concentrations (0 nM, 20 nM, 50 nM, 75 nM, 100 nM, 180 nM, 250 nM and 500 nM) of fluorescence- labeled PRP6-HO7 using the ReadiLink 492 / 516 Antibody Labeling Kit (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions. After washing with FACS buffer and staining with LIVE / DEAD™ fixable Near-IR Dead Cell dye (Invitrogen, ThermoFisher), fixed cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson) and FlowJo software (Flowjo LLC).

[0293] CD36 expression

[0294] Purified primary monocytes were plated at 2.5 x 105cells / 500 pl in 24-well culture plates (Jet Biofil) and differentiated to Mo-macrophages. At the transcriptional level, total RNA from untreated monocytes, and untreated, C4BP(0+) (12 nM)-, C4BP(P~) (12 nM)-, or PRP6-HO7 (32 nM)-treated Mo-macrophages at days 1, 2, 4 and 6 of differentiation, was extracted using Maxwell RSC simply / RNA cells kit (Promega, Madison, WI). Reverse transcription and RT-qPCR was performed using the TaqPath™ 1-Step RT-qPCR Master Mix and the specific CD36 TaqMan Gene Expression Assay (all from Applied BioSystems, Waltham, MA). Quantification was carried out through the AACt method using PPI A as a reference transcript. The relative fold change (FC) was calculated with the equation 2’AACt, normalizing by the monocyte basal (day 0) expression.

[0295] At the translational level, CD36 surface expression was analyzed by flow cytometry (FACS Canto II (Becton Dickinson)) on freshly purified monocytes and after 1, 2, 3 and 6 days of Mo-macrophage differentiation, using the APC-conjugated antihuman CD36 monoclonal antibody (FA6.152) (Beckman Coulter).

[0296] ELISAs

[0297] To confirm and characterize the specificity of C4BP(0-) / PRP6-HO7-CD36 binding the inventors set up direct enzyme-linked immunosorbent assays. Briefly, the inventors immobilized hCD36-Fc, hlgGlK-Fc, hCD36Ll-Fc, mCD36-Fc, hLRPl II-Fc or hLRPl IV-Fc (all from R&D Systems) (1 pg / 100 pl / well) dissolved in coating buffer (100 mM Na2CCh / NaHCCh, pH 9.6) by direct adsorption in a 96-well plate overnight at 4 °C. After three washes with Tris-Tween (50 m Tris pH 7.4 + 150 mM NaCl + 0.2% Tween 20), the plate was blocked with Tris-Tween + 0.5% BSA (or Tris-Tween + 0.5% non-fat dry milk for coated wells with hlgGlK-Fc, hLRPl II-Fc and hLRPl IV- Fc) (100 pl / well) for 1 h at RT. C4BP(0-) and PRP6-HO7 were diluted at 32 nM (or 10 nM for hLRPl binding) in Tris-Tween containing 1 mM CaCL and incubated in duplicate to the plate (100 pl / well) for 2 h at RT. Next, the plate was washed with TrisTween + ImM CaCL and C4BP(0-) or PRP6-HO7 binding was detected by incubation with an anti-C4BP a-chain (PK9008) polyclonal antibody (Kask et al. (2004) J. Biol. Chem. 279: 23869-73) (1:2,000 dilution, in Tris-Tween + 0.5 % BSA, or 0.5 % non-fat dry milk) for 1 h at RT followed by incubation with a polyclonal goat anti-rabbit IgG HRP-conjugated (DAKO P0448; Glostrup, Denmark) (1:2,000 dilution) for 1 h at RT. Alternatively, PRP6-HO7 was also detected by THE™ NWSHPQFEK Tag monoclonal antibody (a-Strep-Tag II) (GenScript) (1:2,500 dilution) followed by incubation with a polyclonal goat anti-mouse IgG HRP-conjugated (DAKO P0447) (1:2,000 dilution). Washing between steps was performed with Tris-Tween. Finally, the plate was developed with TMB (3, 3’, 5, 5’ -tetramethylbenzidine) HRP substrate and stopped with sulfuric acid (IN H2SO4). The absorbance at 450 nm was measured using a Victor X5 2030 Multilabel Reader (PerkinElmer, Waltham, MA). In another set of experiments, with the aim of preventing binding to immobilized CD36, PRP6-HO7 was preincubated with hCD36his (ACROB io systems, Newark, DE) (25 pg / ml) in Tris-Tween + ImM CaCL at RT for 15 min followed by a further 15 min incubation at 4 °C on a rotator before being added to the coated hCD36-Fc wells.

[0298] Concentrations of human TNF-a, and IL-6 were determined from DC supernatants treated with the C4BP(0-), PRP6-HO7, or the analyzed biologies, using the respective DuoSet ELISA kits (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

[0299] Dil-OxLDL uptake

[0300] Dil-OxLDL uptake was analyzed by flow cytometry. Monocytes were plated at 2.5 x 105cells / 500 pl in 24-well UpCell temperature responsive surface plates (Nunc, ThermoFisher, Waltham, MA), in complete medium supplemented with GM-CSF (650 lU / ml) (Gentaur) at 37 °C under 5% CO2. C4BP(0+), C4BP(0-), PRP6-HO7, PRP6-NO, and a-hCD36 Ab and its isotype control a-hlgGlic Ab (both from Stemcell, Vancouver, Canada), and sulfo succinimidyl oleate (SSO) (Cayman Chemical, Ann Arbor, MI) were added to differentiating monocytes at the indicated concentrations. At 16 h later, cells were incubated with Dil-OxLDL (Invitrogen, ThermoFisher) at 2.5 pg / ml for 3 h.

[0301] Cells were collected, washed with PBS, and fixed with 150 pl of 4% paraformaldehyde. The inventors gated the cells according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Labeling with LIVE / DEAD™ fixable Near-IR Dead Cell Stain kit (Invitrogen, ThermoFisher) was also employed to assess their viability. Cells were analyzed using a MoFlo ASTRIOS cytometer (Beckman Coulter, Brea, CA). Subsequent analyses used FlowJo software (Flowjo LLC, Ashland, OR).

[0302] Monocyte surface CD36 expression in inflammatory bowel disease (IBD) patients vs. control individuals

[0303] IBD patients derived to the Gastroenterology Unit from Bellvitge University Hospital for routine colonoscopy underwent protocol blood extraction under informed written consent at hospital admission (study was approved by the Ethics Committee from the Bellvitge University Hospital Ref.- PR323 / 20, in accordance with institutional guidelines and the Declaration of Helsinki). The PBMCs were isolated less than 16 h after extraction through FicolLPaque density centrifugation (GE Healthcare BioSciences AB) as described for healthy donors. In both cases, cell-surface expression of CD36 on monocytes was analyzed using the APC-conjugated anti-human CD36 monoclonal antibody (FA6.152) (Beckman Coulter). The inventors gated the CD14+cells according to FSC and SSC parameters and with FITC-conjugated anti-CD14 (TUK4) (Miltenyi Biotec). Labeling with LIVE / DEAD™ fixable Near-IR Dead Cell Stain kit (Invitrogen, ThermoFisher) was also employed to assess their viability. Stained and fixed cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson). Subsequent analyses used FlowJo software (Flowjo LLC).

[0304] A375 cell culture and pro-inflammatory induction

[0305] A375 melanoma cells were seeded in wells of 24- well UpCell temperature- responsive surface plates (Nunc, ThermoFisher) at 4 x 104cells / 500 pl DMEM High Glucose medium (Sigma Aldrich-Merck, Darmstadt, Germany) supplemented with 100 mg / ml streptomycin, 100 lU / ml penicillin, 2 mM L-glutamine (all from Gibco, ThermoFisher) and 10% heat-inactivated FBS (Life technologies, ThermoFisher). Cells were induced by 24 h incubation with the pro-inflammatory stimulus TNF-a (Peprotech, Thermo fisher) + IFN-y (Miltenyi Biotec) (both at 100 ng / ml), and at 48 h after induction CD36 expression was analyzed by flow cytometry using the APC-conjugated anti-human CD36 MoAb (FA6.152) (Beckman Coulter). APC-conjugated anti-IgGlk (Beckman Coulter) was used as the corresponding isotype control. Cells were gated according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Staining with LIVE / DEAD Fixable Near IR dye (Invitrogen, Thermofisher) was also employed to assess their viability. Stained cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson). Subsequent analyses used FlowJo software (Flowjo LLC).

[0306] Immunomodulatory activities of C4BP( -), PRP6-HO7, anti-CD36 Abs and biologies

[0307] CD14+primary monocytes were obtained as previously described. Monocytes were plated at 2.5 x 105cells / 500 pl in 24-well culture plates (Jet Biofil), in complete medium at 37 °C under 5% CO2. Mo-DCs or Mo-macrophages were generated as described in a previous section. C4BP(0-), PRP6-HO7, antibodies (monoclonal IgGl anti-human CD36 (FA6-152; Stemcell), monoclonal IgA anti-human CD36 (JC63.1; Cayman Chemical), and monoclonal IgM anti-human CD36 (SMo; Bio-Rad)), and biologies (Etanercept (Sigma Aldrich-Merck), Infliximab and Abatacept (both from MedChemExpress, South Brunswick Township, NJ) were added at day 0 to differentiating monocytes, all at 32 nM for 24 h. For DC maturation, Mo-DCs, either untreated or treated, were further stimulated for 48 h with 5 pg / ml LPS (Escherichia coli 055.B5, Sigma Aldrich-Merck) at day 5. For Ml polarization, Mo-macrophages (M0) were stimulated at day 6 for 48 h with 20 ng / ml LPS (Escherichia coli 055.B5, Sigma Aldrich-Merck) plus 20 ng / ml INF-y (Miltenyi Biotec). Cell-surface phenotypes were analyzed using the following MoAbs: APC-conjugated anti-CD64 (10.1.1), APC- conjugated anti-CD83 (REA714), PE-conjugated anti-CD86 and PE-conjugated anti- CD80 (REA661) (Miltenyi Biotec). Thus, after washing with PBS, cells were subsequently stained with the respective MoAbs, according to the manufacturer’s instructions, in 60 pl PBS for 20 min at RT. Staining was stopped by addition of 150 pl fixation buffer (PBS + 4% paraformaldehyde). Cells were gated according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Labeling with LIVE / DEAD™ fixable Near-IR Dead Cell Stain kit (Invitrogen, ThermoFisher) was also employed to assess their viability. Stained and fixed cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson). Subsequent analyses used FlowJo software (Flowjo LLC).

[0308] Mice, study design and follow up

[0309] Animals had free access to food and water and were maintained under standard laboratory conditions, at 20-24 °C and 40-70% relative humidity, with 12-hour fluorescent light / 12-hour dark cycle. 12- week old female (n = 6) Apolipoprotein E- deficient mice (ApoE(- / -)) (Charles River, Wilmington, MA) were fed a high-fat diet (Clinton / Cybulsky High Fat Rodent Diet With Regular Casein and 1.25% Added Cholesterol, Cat.No.D12108C, Research diets, Inc.) until the end of the experiment. To assess the therapeutic potential of C4BP(0-) in the ApoE(- / -) model, 100 pg / mouse of C4BP(0-) was administered subcutaneously, twice a week, from week 27 to week 31 (4 weeks). A control group underwent vehicle PBS administration following the same administration route and schedule as did the C4BP(0-) group in the study.

[0310] Livers were dissected and processed for histological analysis. Liver slices were fixed in 4% paraformaldehyde and embedded in paraffin. Standard histochemical analysis was performed in 5 pm sections stained with hematoxylin / eosin. To visualize lipid deposits, liver slices were also fixed in 4% paraformaldehyde and embedded in Tissue Tec OCT compound (Sakura, Alpen aan den Rijn, Netherlands) and stored at -80 °C. Cryostat sections (5pm) were stained with oil red O solution. Stained sections were visualized in a Nikon Eclipse 80i microscope (Nikon Instruments, Tokyo, Japan).

[0311] All experiments were carried out in accordance with the current EU legislation on animal experimentation and were approved by “CEE A: Animal Experimentation Ethics Committee”, the Institutional Ethics University of Barcelona Committee for Animal Research, and the Generalitat de Catalunya (DARP: 8520).

[0312] Statistical analysis

[0313] Statistical analyses and graphic visualization of the data were performed using the GraphPad Prism 6.0 software (GraphPad software, Inc, La Jolla, CA). Significance in the ELISA assays were inferred through the non-parametric Mann- Whitney U test. Repeated measures one-way ANOVA, corrected for multiple comparisons using Dunnett’s method was performed to contrast median fluorescence intensity (MFI) under different experimental conditions with respect to a reference condition in the PRP6- HO7 cell binding assays, Dil-OxLDL uptake assays, and immunomodulatory activity assays. Regarding the saturation binding curve, fitting was carried out by non-linear regression using saturation binding - one site specific binding with Hill Slope equation (GraphPad Prism 6.0). The time courses of CD36 transcript and surface protein expression through Mo-macrophage differentiation were analyzed by two-way ANOVA (or bidirectional analysis of variance) corrected for multiple comparisons using the Holm-Sidak method. To compare monocyte surface CD36 MFI values between IBD patients and healthy individuals, the unpaired t-test was employed. Finally, to analyze the difference of CD36 MFI between induced and non-induced A375 cells, the inventors employed the non-parametric Wilcoxon matched pairs test. Data are expressed as mean values ± SD. In all cases, a p-value < 0.05 was considered significant.

[0314] Example 10

[0315] Materials and methods

[0316] Monocyte culture, differentiation, and treatment

[0317] Total blood from both the healthy and the CD36-deficient donors was obtained from the Blood and Tissue Bank (Barcelona, Spain), their PBMCs were isolated at less than 16 h after extraction through Ficoll-Paque density gradient centrifugation (Cytiva, Marlborough, MA) and plated at 8.0x105cells / 500 pl in 24-well culture plates (Jet Biofil, Guangzhou, China), in RPMI 1640 (Gibco, ThermoFisher, Waltham, MA) supplemented with 100 mg / ml streptomycin, 100 lU / ml penicillin, 2 mM L-glutamine (all from Invitrogen, Carlsbad, CA) and 10% heat- inactivated FBS (Life technologies, ThermoFisher) (complete medium) at 37 °C under 5% CO2. Monocyte cultures were supplemented with GM-CSF (800 lU / ml) plus IL-4 (500 lU / ml) (both from Gentaur, Kampenhout, Belgium) for monocyte to monocyte- derived dendritic cell (Mo-DC) differentiation.

[0318] C4BP(0+), C4BP(0-) (both at 5 pg / ml) or PRP6-HO7 (at 3 pg / ml) were added on day 0 to DC-differentiating monocytes for 24 h. For DC maturation, on day 5 immature DCs (iDC), either untreated or treated with the different proteins, were further stimulated for 48 h with 5 pg / ml LPS (Escherichia coli 055. B5, Sigma Aldrich, Merck, Darmstadt, Germany)

[0319] CD36 expression

[0320] CD36 surface expression was assessed on CD14+monocytes using FITC- conjugated anti-CD14 (TUK4) (Miltenyi Biotec, Bergisch Gladbach, Germany) with APC-conjugated anti-human CD36 monoclonal antibody (FA6.152) or its corresponding APC-conjugated IgGl isotype control (both from Beckman Coulter, Brea, CA). Briefly, after washing with PBS, cells were subsequently stained with the respective MoAbs, according to the manufacturer’s instructions, in 60 pl PBS for 20 min at room temperature. The staining was stopped by cell resuspension in 150 pl fixation buffer (PBS + 4% paraformaldehyde). We gated the cells according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Labeling with LIVE / DEAD™ fixable Near-IR Dead Cell Stain kit (ThermoFisher) was also employed to assess their viability status. Stained and fixed cells were analyzed using a FACSCanto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ). Subsequent analyses used FlowJo software (Flowjo LLC, Ashland, OR).

[0321] Mo-DC cytokine secretion

[0322] Concentrations of human TNF-a, IL-6, CCL4 and IL-10 were determined from Mo-DC supernatants either untreated or treated with the C4BP isoforms or PRP6-HO7 using the respective DuoSet Elisa kits (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

[0323] Examples 11-13

[0324] Therapeutic potential of PRP6-HO7 in a rat model of acute ulcerative colitis

[0325] Materials and methods

[0326] DSS-induced colitis model The acute colitis model was established in male Sprague-Dawley rats (Charles River Laboratories, Wilgminton, MA) weighting around 180-200 g (6 weeks old) through the administration of DSS (35-50 kDa; TdB Consultancy, Uppsala, Sweden) at a concentration of 5% (w / v) into the drinking water ad libitum for 7 days (freshly prepared every 2-3 days) after which the DSS solution was replaced by fresh water until the sacrifice, on day 10.

[0327] Rats were treated subcutaneously with PRP6-HO7 at a concentration of 2.5 mg / kg, or with PBS (for the DSS colitis control group) on day 5. In addition, a reference treatment group was treated daily from days 0 to 9 with 25 mg / kg of Mesalazine (Sigma-Aldrich, St Louis, MO) through oral gavage. A further untreated control group (blank) received normal tap water throughout the study.

[0328] During the experimental period, rats were housed in a controlled environment with free access to a standard diet. The severity of colitis was evaluated daily by monitoring body weight and other clinical manifestations such as stool consistency and bleeding. These parameters were registered as the disease activity index (DAI) based on the scoring system previously defined by Martin et al. (2016) Methods Mol Biol. 1371: 197-203 (Table 5). Occult blood was assessed using the hemoCARE Slide Test for faecal occult blood (CARE Diagnostica, Osterreich, Austria).

[0329] Table 5. Disease activity index (DAI) scoring system to assess the severity of colitis in the rat DSS-induced colitis model

[0330] Score Body weight loss (%) Stool consistency Bleeding

[0331] 0 0 Normal No blood

[0332] 1 1-5 Normal, but pale Occult bleeding

[0333] 2 5-10 Soft Visible in stools

[0334] 3 10-15 Loose / mild diarrhea Visible in and around stools

[0335] 4 >15 Watery diarrhea Fresh rectal bleeding

[0336] DAI: the sum of the score in each category (i.e., score of body weight + stool consistency + bleeding) On day 10, rats were anesthetized with isoflurane, euthanized, and their colons were extracted, measured, and weighed. From both proximal and distal regions, fragments were collected, stored at 4°C in 4% paraformaldehyde for further histological analyses. The remaining fragment was stored for 48h in MACS Tissue Storage solution (Miltenyi Biotec, Bergisch Gladbach, Germany).

[0337] Histomorphological analysis

[0338] Transversal colon segments were fixed in 4% paraformaldehyde and embedded in paraffin. Standard histochemical analysis was performed in 5 pm sections stained with haematoxylin / eosin. Tissue sections were visualized in a Zeiss Axio Observer Z1 inverted microscope equipped with a structured illumination Apotome system with the Axiocam 208 colour camera (Carl Zeiss, Oberkochen, Germany). The digital processing was performed with Zen Blue 3.0 software (Carl Zeiss). Scale bar tiled images: 200pm. Scale bar crop images: 100pm.

[0339] The histomorphological evaluation of intestinal inflammation was based on a standardized score system (Adapted from Erben et al. Int J Clin Exp Pathol 2014;7(8):4557-4576) (Table 6) considering the inflammatory cell infiltration, epithelial changes such as crypt hyperplasia and Goblet cell loss, and the overall mucosal architecture.

[0340] Table 6. Standarized scoring system for histomorphological evaluation in the rat DSS- induced colitis model

[0341] Inflammatory cell infiltrates

[0342] 0 Minimal <10%

[0343] 1 Mild to moderate 10-50%

[0344] 2 Marked >50%

[0345] Epithelial changes

[0346] Hyperplasia

[0347] 0 None

[0348] 1 Mild to moderate

[0349] 2 Marked Goblet cell loss

[0350] 0 None

[0351] 1 Mild to moderate

[0352] 2 Marked

[0353] Mucosal architecture

[0354] 0 Normal

[0355] 1 Irregular crypts (focal architecture loss, blunted crypts)

[0356] 2 Crypt and epithelium loss

[0357] Final score corresponds to the sum of the values of the different histomorphological parameters evaluated

[0358] Lamina propria cell isolation and flow cytometry

[0359] Colons were extracted and washed with cold PBS to remove feces, and the middle section (~2cm) was kept for 48h in MACS Tissue Storage solution (Milteny Biotec). These sections were opened longitudinally and cut into small fragments (<lmm) followed by an incubation with a digestion solution containing collagenase (375 pg / ml) (Sigma-Aldrich) in pre-warmed 0.1% BSA in PBS for 45 min at 37°C with agitation, pipetting every 5 min to disaggregate the tissue. After stopping the digestion with cold 0.1% BSA in PBS the samples were centrifuged, resuspended and filtered through a 100pm strainer. The samples obtained were then resuspended in staining solution (1% BSA in PBS).

[0360] After blocking the antibodies’ nonspecific binding to Fc receptors using Purified Mouse Anti-Rat CD32 (D34-485) (BD Biosciences, Franklin Lakes, NJ) at 4°C for 5 min, lamina propria cells were incubated for 20 min at room temperature with the following antibodies: PEVio770-conjugated anti-CD45 (REA504), APC-conjugated anti-CD3 (REA223), PE-conjugated anti-CDl lb / c (REA325) and FITC-conjugated anti-Granulocytes (REA535) (all of them from Milteny Biotec). Labelling with LIVE / DEAD™ fixable Near-IR Dead Cell Stain kit (ThermoFisher, Waltham, MA) was also employed to assess their viability status. Cells were finally washed and resuspended in 0.1% BSA in PBS and filtered with a 100 pm filter. We gated the cells according to forward scatter (FSC) and side scatter (SSC) parameters to exclude debris. Stained cells were analysed using a FACSCanto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ). Subsequent analyses used FlowJo software (Flowjo LLC, Ashland, OR).

[0361] Statistical analysis

[0362] Statistical analyses and graphic visualization of the data were performed using the GraphPad Prism 6.0 software (GraphPad software, Inc, La Jolla, CA). Body weight and DAI parameters were analyzed by two-way ANOVA. The area under the curve (AUC), the colon weight / length ratio and the histomorphological scores were assessed by one-way ANOVA. Both ANOVAs were corrected for multiple comparisons using Dunnett’s method. Different experimental conditions were compared with the DSS colitis control group. Data are expressed as mean values ± SEM. In all cases, a p-value < 0.05 was considered significant.

[0363] Example 1-Putative receptor screening for the orphan ligands C4BP( -) / PRP6- HO7 in myeloid cells

[0364] PRP6-HO7 is a new C4BP(0-) analogue, an oligomeric form including only the complement control protein 6 (CCP6) domain of the C4BP a-chain, which is sufficient to “reprogram” myeloid cells (monocytes differentiating to DCs or to macrophages, and neutrophils), from a pro-inflammatory and immunogenic phenotype to an antiinflammatory and tolerogenic state.

[0365] The inventors have shown that C4BP(0-), acting directly over DC- or macrophage-differentiating monocytes conferred a strong immunomodulatory activity in these cells, characterized by a hypo response to pro-inflammatory stimuli such as LPS (Olivar et al. (2013) J. Immunol. 190: 2857-72). To further assess the surface receptor responsible for transducing this immunomodulatory activity the inventors employed: 1) PRP6-HO7 as a ligand (bait), a recombinant curtailed version of C4BP(0- ) holding only the immunomodulatory activity but not the complement inhibitory activity (Serrano et al. (2022) Front. Immunol. 13: 883743). This would increase the specificity of the screen on DC-differentiating monocytes induced by GM-CSF and IL- 4; and 2) Unbiased proteomic analysis through ligand-based receptor capture (LRC) on living cells under near-physiological conditions using TriCEPS, a trifunctional chemoproteomic reagent (Frei et al. (2012) Nat. Biotechnol. 30: 997-1001; Frei et al. (2013) Nat. Protoc. 8: 1321-36).

[0366] Thus, PRP6-HO7 was coupled with the LRC-TriCEPS crosslinker and incubated with DC-differentiating monocytes. In parallel, the inventors used TriCEPS quenched with glycine (Gly) as a negative control. TriCEPS binds covalently to the glycans of random surface proteins but also the targets of the ligands. Due to the ligand binding, the target proteins are more heavily labeled compared to the random surface proteins. The crosslinked proteins are purified and processed for analysis by mass spectrometrybased proteomics (LC-MS / MS). The relative quantification between PRP6-HO7 and glycine enabled the identification of captured CD36 as candidate target (Figure 1).

[0367] A total of 262 membrane associated proteins were identified by LC-MS / MS and used for quantitation and statistical analysis. Three proteins were significantly enriched in the ligand of interest, PRP6-HO7, against the control (Gly samples) and are therefore suggested as candidates to interact with PRP6-HO7 protein. One of them, labeled as C4BPA, corresponds to the PRP6-HO7 ligand used as bait, 100% homologous to the human C4BP a-chain. From the remaining two protein hits, the corresponding to the CD36 receptor (log2FC = 1.74; adj p value = 2.72 x 10“4) was further explored as a potential PRP6-HO7 ligand.

[0368] Example 2- PRP6-HO7 binds CD36 with high avidity

[0369] A preliminary assay was performed to assess the interaction of PRP6-HO7 in the surface of differentiating monocyte-derived dendritic cells (Mo-DCs). The binding of PRP6-HO7 to the putative Mo-DC receptor(s) was saturable, cooperative (h = 1.6 + 0.3), and of high affinity (95 + 12 nM), suggesting a receptor clustering effect induced by the heptavalent PRP6-HO7 interaction (Figure 2).

[0370] Next, the inventors employed surface plasmon resonance to confirm the above LRC results and to infer the specific binding kinetics and affinity analysis of PRP6-HO7 to CD36. Thus, the inventors immobilized a recombinant chimeric polypeptide including the whole extracellular domain of human CD36 fused to the IgGl Fc domain (hCD36-Fc) in a sensor chip and analyzed its binding to PRP6-HO7 at concentrations ranging from 15 nM to 1.0 pM. The obtained sensorgrams confirmed specific binding of PRP6-HO7 to CD36, with binding response vs time curves that were best fit to a two- state reaction model (%2= 0.159) approaching multivalent binding avidity, with an estimated KD = 32.2 n (Table 7 and Figure 3), although the real KD might be lower because of the premature steady-state evaluation (plots not flatten out sufficiently to reach steady state).

[0371] Table 7. Kinetic parameters for PRP6-HO7 binding to CD36 ka 104kd IO’2kal 104kdl IO’2ka2 IO’2kd2 IO’3KD Rmax ' / 2

[0372] (MV1) (S’1) (MV1) (S’1) (S’1) (S’1) (nM) (RU) (RU2)

[0373] 1:1 Binding 1.817 0.073 . . . . 40.2 69,97 0,396

[0374] Bivalent binding - - 0.754 0.126 0.010 0.002 - 84,31 0,330

[0375] Two-state reaction - - 2.646 2.251 2.841 1.118 32.2 80,39 0,159

[0376] This model has been successfully applied to FGPI (apoH) binding to phospholipid membranes (Gamsjaeger et al. (2005) Biochem. J. 389: 665-673), apoE binding to heparan sulfate proteoglycans (Futamura et al. (2005) J. Biol. Chem. 280:5414-22), apoA-I binding to HDL (Lund-Katz et al. (2010) J. Lipid Res. 51: 606- 17), or Shiga toxin (Stx) binding to its receptor glycolipid Gb3Cer (Nakajima et al. (2001) 276: 42915-22. Interestingly, Stxs have a subunit composition in which 5 x 7.7 kDa B subunits are associated noncovalently with a globular 32 kDa A subunit through its carboxy-terminal amino acids (Lee et al. (2016) Toxins 8: 77), a structure similar to PRP6-HO7. Furthermore, upon Stxs binding to the cell surface, the receptor-ligand complex is then internalized by endocytosis mediated by clathrin-coated pits (Bergan et al. (2012) Toxicon 60: 1085-107). Analogously, an initial interaction of one of the CCP6 domains of PRP6-HO7 with a CD36 receptor molecule would lead to the subsequent heptavalent PRP6-HO7 sequential binding of the remaining CCP6 domains to additional CD36 receptor molecules and its clustering. In fact, the kaivalues of the first interaction step were significantly higher than those obtained for the second one (by a factor of approx. 106). Moreover, the low kd2 rates determine the flat course of the final dissociation part of the sensorgrams. Consequently, the formation of a stable PRP6-HO7-CD36 complex might trigger internalization. To note, it has been recently reported that CD36 binding to diverse CIDRa2-6 domains of PfEMPl proteins from the malaria parasite Plamodium falciparum also adopt nanomolar interactions with slow off-rates, interfering with lipoprotein particle interaction (Hsieh et al. (2016) Nat. Commun. 7: 12837). Furthermore, it has been shown that the calcium-binding protein S100A12 (calgranulin C), which can also form hexamers, binds CD36 with nanomolar affinity in a binding site shared by thrombospondin or collagen and with a two-state reaction kinetics (Tondera et al. (2017) Amino Acids 49: 183-91).

[0377] Finally, the inventors confirmed the specificity of PRP6-HO7 binding to CD36 in differentiating Mo-DCs by flow cytometry. Thus, specific binding of Strep-Tag II- tagged PRP6-HO7 to the Mo-DC surface over the background fluorescence given by incubation with the secondary FITC-conjugated anti-Strep-Tag II antibody alone, could be blocked by pre-incubation of these cells with an anti-CD36 MoAb, but not with the corresponding isotype control Ab IgGik (Figure 4).

[0378] Example 3- Specificity of PRP6-HO7 and C4BP(0-) binding to CD36

[0379] To substantiate the above results, the inventors turned on enzyme-linked immunosorbent assays (ELISA) as reliable in vitro ligand-binding systems. First, the inventors verified significant PRP6-HO7 binding to immobilized hCD36-Fc assessing complex formation through an anti-Strep-Tag II MoAb. Competition of PRP6-HO7- CD36-Fc binding was essentially abolished by competition with a soluble CD36 receptor (hCD36his). Moreover, immobilized hCD36-Fc isotype control MoAb (hlgGik) was unable to bind PRP6-HO7 (Figure 5A). These results were confirmed by substitution of the anti-Strep-Tag II MoAb by a polyclonal anti-C4BP-a Ab (PK9008) as Ab for detection. Moreover, using this same PK9008 Ab, the inventors also demonstrated specific binding of C4BP(0-) to hCD36-Fc, although the magnitude of the C4BP(P-)-hCD36-Fc interaction was smaller compared to the PRP6-HO7-hCD36-Fc interaction (Figure 5D). This suggests steric restrictions due to the almost 5 times larger size of C4BP(P~) with respect to PRP6-HO7. Furthermore, both PRP6-HO7 and C4BP(P~) interacted distinctively with hCD36 (SCARB3) and were not able to interact with its close lipid scavenger receptor family homologue SR-B1 (SCARB1 or CD36L1) (Figures 5B and 5E).

[0380] On the other hand, PRP6-HO7 was also able to bind immobilized mouse CD36- Fc albeit lower intensity that human CD36-Fc (Figures 5C and 5F). In fact, human and mouse CD36 share more than 80 % homology. Nevertheless, through ELISA assay the inventors were unable to demonstrate binding between C4BP(0-) and mouse CD36-Fc, although their previous in vivo assays have suggested a functional interaction between both molecules. Finally, LRP1, an endocytic receptor involved in the catabolism of various heparin-binding proteins, has been suggested as a putative C4BPa receptor through the interaction of domains II and IV with the heparin-binding site of C4BPa (CCP1-CCP3) (Western et al. (2002) J. Biol. Chem. 277: 2511-6; Spijkers et al. (2008) Eur. J. Immunol. 38: 809-17). Accordingly, the inventors reproduced the binding of C4BP(0-) to domains II and IV of LRP1 but, conversely, the binding of PRP6-HO7 to these LRP1 domains was negligible (Figure 6).

[0381] Example 4- PRP6-HO7 modulates CD36 expression in differentiating monocytes

[0382] CD36 expression is high in human monocytes / macrophages, according to its fundamental role in key biological processes such as innate immunity, inflammation, and atherosclerosis (Chen et al. (2022) J. Exp. Med. 219: e20211314). The inventors set up to assess the time course of CD36 expression in monocyte to macrophage (M0) differentiation in culture by exposure to GM-CSF. At transcriptional level, CD36 mRNA expression was downregulated at the beginning of monocyte (CD14+) differentiation (day 1), recovering progressively up to M0 (days 2 to 6), but remaining slightly below the initial level observed in monocytes. Both C4BP(0-) and PRP6-HO7 could significantly downregulate CD36 expression. Nevertheless, PRP6-HO7 induced a stronger downregulation, which persisted until the end of the differentiation to M0 (FC = 16) (Figure 7A). Analogously, at the translational level CD36 surface receptor expression was found high in monocytes, but differentiation to M0 induced its downregulation until day 1 and subsequently underwent a progressive upregulation, reaching a level at M0 like that initially shown for monocytes. Again, both C4BP(0-) and PRP6-HO7 significantly downregulated surface CD36 expression, which remained up to 2-fold below that of untreated M0 macrophages at the end of the differentiation (Figure 7B). In any case, C4BP(0+), lacking immunomodulatory activity, was incapable to downregulate CD36 expression.

[0383] Altogether, C4BP(0-) and PRP6-HO7 interaction with the CD36 receptor, besides signaling and / or blocking activities on differentiating monocytes, also induces a significant downregulation of this multifunctional receptor, which could work as a safety check for undesired or prolonged functions of the C4BP(P-) / PRP6-HO7 - CD36 complex. Example 5- PRP6-HO7 and C4BP(0-) interfere with the oxLDL-CD36 axis in differentiating monocytes and macrophages

[0384] Blood levels of oxidized low-density lipoprotein (oxLDL) rise with age and upon ingestion of high cholesterol diet, which increases the risk of metabolic syndrome, cardiovascular disease, and acute myocardial infarction. Moreover, oxLDL levels may also be elevated in patients with autoimmune disorders (Frostegard J et al. (2005) Arthritis Rheum. 52: 192-200). Macrophage CD36 participates in atherosclerotic arterial lesion formation through its interaction with oxLDL, which triggers signaling cascades for inflammatory responses. CD36 functions as scavenger receptor in oxLDL uptake and foam cell formation, which is the initial critical stage of atherosclerosis. In fact, CD36 deficiency reduces atherosclerotic lesion formation. Moreover, oxLDL involves an increase in the responsiveness of monocytes (amplification of pro-inflammatory cytokine production, ROS production, epigenetic reprogramming, and metabolic activation) (Bekkering et al. (2016) Clin. Vaccine Immunol. 23: 926-33).

[0385] Thus, the inventors set out to assess whether PRP6-HO7, through binding CD36, was able to interfere with oxLDL uptake in macrophage-differentiating monocytes. In fact, PRP6-HO7 prevented the uptake of oxLDL with analogous efficiency that the known CD36 irreversible inhibitor sulfo-N-succinimidyl oleate (SSO) (Kuda et al. (2013) J. Biol. Chem. 288: 15547-55). In comparison, a blocking anti-CD36 Ab also reduced oxLDL uptake in these cells, albeit with a reduced efficiency (Figure 8).

[0386] Moreover, the PRP6-HO7 precursor molecule C4BP(0-), but neither its homologous non-immunomodulatory isoform C4BP(0+) not the PRP6-HO7 monovalent analogue PRP6-NO, was also able to completely block oxLDL uptake in macrophage-differentiating monocytes (Figure 9).

[0387] This suggests that the hexavalent interaction of the CCP6 domain from both C4BP(b-) and PRP6-HO7 with CD36 is a necessary and sufficient condition to prevent oxLDL uptake in macrophage differentiation.

[0388] Example 7- CD36 is overexpressed in inflammatory conditions

[0389] The inventors have found the scavenger receptor CD36 significantly overexpressed in monocytes from auto immune / auto inflammatory disease patients, such as in IBD patients, compared to those from healthy individuals (Figure 10A). In addition, CD36 expression seems to be induced by pro-inflammatory cytokines (Figure 10B and C). In fact, CD36 has been reported to be overexpressed or induced in a variety of immune-inflammatory conditions such as atherosclerosis and hyperglycemia (Lopez-Carmona et al. (2017) Diabetol. Metab. Syndr. 9: 55), NAFLD, a variety of cancers such as hepatocellular carcinoma (Luo et al. (2021) Cell Death Dis. 12: 328), and other pathological drivers such as epithelial to mesenchymal transition (Nath et al. (2015) Sci. Rep. 5: 14752; Deng et al. (2019) J. Transl. Med. 17: 352), to name only a few. Moreover, forced overexpression of CD36 accelerates the progression of inflammatory diseases (Jung et al. (2018) Kidney Res. Clin. Prac. 37: 30-40).

[0390] Example 8- CD36-mediated immunomodulation is induced by PRP6-HO7 but neither by anti-CD36 Abs nor by anti-inflammatory chimeric Abs or proteins

[0391] To compare the immunomodulatory activity of C4BP(0-) and PRP6-HO7 versus different antibody isotypes against CD36, specifically IgG, IgA, and IgM, or versus TNF-a blockers Infliximab (chimeric anti-TNF-a Mo Ab) and Etanercept (fusion protein between TNFR2 and the Fc of IgGl), or the CD80 / CD86 blocker Abatacept (fusion protein between the extracellular domain of CTL-4 and the Fc of IgGl) the inventors incubated all of them with differentiating monocytes at the same molar concentration (32 nM) and assessed pro-inflammatory surface marker levels in differentiated (CD64), and Ml (LPS + IFN-y) polarized (CD80) Mo-macrophages, and in LPS-matured Mo-DCs (CD83 and CD86). As previously reported (Olivar et al. (2013) J. Immunol. 190: 2857-72, Serrano et al. (2022) Front. Immunol. 13: 883743), significant immunomodulation was induced by both C4BP(0-) and PRP6-HO7 in Mo- DCs. The same tendency was observed for Mo-macrophages, although it did not reach statistical significance due to the low number of samples tested. Conversely, neither the anti-CD36 Abs tested (Figure 11) nor the TNF-a blockers Infliximab and Etanercept, or the CD80 / CD86 blocker Abatacept (Figure 12) could reduce the inflammatory surface marker and / or pro-inflammatory cytokine levels in both Mo-DCs and Mo- macrophages.

[0392] Example 9- C4BP(0-) prevents the development of hepatic steatosis in ApoE(- / -) mice fed with a high-fat diet

[0393] CD36 is a key driver of NAFLD pathogenesis (Rada et al. (2020) Cell Death Dis. 11: 802). Moreover, CD36-deficient mice are resistant to both alcohol- and high- carbohydrate-induced hepatic steatosis (Clugston et al. (2014) J. Lipid Res. 55: 239-46). The inventors set up to analyze the effect of C4BP(0-) abrogating steatosis in the livers of hyperlipidemic (ApoE(- / -)) mice fed with a Western diet for 19 weeks. Compared with control PBS-treated ApoE(- / -) mice, subcutaneous administration of C4BP(0-) in these mice in the last 4 weeks prevented the development of hepatic steatosis (lipid accumulation in the form of lipid droplets) evidenced by both hematoxylin and eosin staining and Oil Red O staining of liver sections (Figure 13). This suggests that C4BP(P~) interaction with the CD36 receptor positively influences hepatic lipid homeostasis.

[0394] Example 10-C4BP(P~) and PRP6-HO7 are unable to modulate inflammatory cytokine / chemokine secretion in CD36-deficient Mo-DCs

[0395] To comparatively assess the immunomodulatory behavior of C4BP(0-) and PRP6-HO7 over PBMCs isolated from a type I CD36-deficient individual, holding compound heterozygous mutations in the CD36 alleles (CD36 c.975T>G and CD36 c.l 57 _158delinsG), the corresponding surface protein was labeled with a fluorescent anti-CD36 antibody. Flow cytometry analysis of CD14+monocytes from such individual (P36 1) validated the absence of CD36 expression in the surface of these cells, compared with the elevated CD36 expression present in monocytes from two normal (control) individuals (STI 3 and ST14) (Figure 14).

[0396] Further, analysis of cell supernatants from Mo-DCs pre-incubated with the major C4BP isoforms (C4BP(0+) and C4BP(0-)) or PRP6-HO7 and matured with the pro- inflammatory stimulus LPS confirmed that only those cells from normal individuals with high surface expression of CD36, but not the individual deficient in CD36 expression, displayed low or null secretion of inflammatory cytokines / chemokines (TNF-a, IL-6, CCL4) and, conversely, increased secretion of the anti-inflammatory cytokine IL- 10 (Figure 15).

[0397] Example 11-PRP6-HO7 ameliorates clinical signs of DSS-induced colitis in rats

[0398] To further evaluate the impact of PRP6-HO7 in acute colitis, the DSS-induced colitis rat model was employed, which closely resembles human colitis pathology compared to the DSS-induced colitis mouse model. In addition, mice lack the CCP6 immunomodulatory domain in its C4BP alpha chain, and also the full C4BP beta chain. Dextran sulphate sodium (DSS) can reproduce the disease by interfering with the intestinal epithelial barrier, resulting in the release of inflammatory mediators. PRP6- HO7 was then administered subcutaneously (2.5 mg / kg) on day 5, once some macroscopic pathological traits were already evident. As a reference, the standard of care anti-inflammatory drug Mesalazine was administered daily via oral gavage (25 mg / kg) during 9 days upon the start of the DSS induction (see experimental design in Figure 16A).

[0399] Body weight evolved positively in all but in the DSS-induced and untreated group, which began to decrease on the 5th day of DSS exposure (Figure 16B). The reduction in body weight was further associated with a drastic increase in diarrhea and fecal blood, which contributed to the DAI score (Figure 16C and 16D). Conversely, PRP6-HO7 treated individuals displayed a significantly reduced DAI progression (p<0.01), while the Mesalazine-treated group failed to do so. The latter exhibited a similar DAI score evolution to the untreated DSS group, with both only showing improvement once the DSS solution was replaced with fresh water on day 7. Variation in the colon weight / length ratio were also assessed in the acute rat model (Figure 17), as a shorter and heavier colon is characteristic of experimental colitis. Both PRP6-HO7 and Mesalazine attenuated the increment in this ratio, which was evident in the DSS- induced only group. Nevertheless, only the PRP6-HO7 group achieved statistical significance. Together these data indicate that PRP6-HO7 treatment diminishes the clinical symptoms of acute colitis in rats.

[0400] Example 12- PRP6-HO7 attenuates colonic damage and histopathological signs of acute DSS-induced colitis in rats

[0401] DSS induced severe alterations in the colonic structure of untreated rats (Figure 18 A, DSS). A significant inflammatory immune cell infiltration was noted in the lamina propria extending to the submucosa layer, along with severe epithelial damage including erosion, crypt loss and hyperplasia. In contrast, only the colonic tissue from PRP6-HO7-treated rats (Figure 18A, PRP6-HO7) was significantly more preserved, its appearance being closer to the healthy group (Figure 1A8, Blank).

[0402] The histological score (Figure 18B) was high in the DSS-induced and untreated group, but it became significantly reduced (p < 0.01) in the PRP6-HO7-treated group. On the other hand, the Mesalazine-treated group, whose response to the treatment varied widely among the individuals, did not exhibit a noticeable reduction in the overall histological score compared to the DSS-induced and untreated group. These results indicate that PRP6-HO7 treatment attenuates colon inflammation and important alterations of the colonic mucosal architecture induced by DSS stimulation. Example 13-PRP6-HO7 reduces colonic lamina propria immune infiltration of acute DSS-induced colitis

[0403] An elevated expression of the cell surface markers CD 11b and GR1, which include monocytes and eosinophils, but mainly neutrophils, was employed to track inflammation within the intestinal lamina propria. Thus, flow cytometry analysis showed that neutrophil cell infiltration was virtually absent in DSS-induced and PRP6-

[0404] HO7-treated rats. In contrast, the DSS-induced and untreated group experienced heavy neutrophil infiltration in their lamina propria (Figure 19). These results further stress the therapeutic potential of PRP6-HO7 on acute DSS-induced colitis.

Claims

CLAIMS1. An oligomer comprising at least three polypeptides, wherein each polypeptide comprises the CCP6 domain of the C4BP alpha chain or a functionally equivalent variant thereof and an oligomerization domain, and wherein said oligomer does not comprise the C4BP beta chain, for use in the prevention and / or treatment of a disease involving an undesired activation of the immune system in a subject having CD36 expression on the surface of the monocytes / macrophages.

2. The oligomer for use according to claim 1, wherein the oligomer comprises at least six polypeptides.

3. The oligomer for use according to any one of claims 1 or 2 wherein the oligomer is an homooligomer.

4. The oligomer for use according to any one of claims 1 to 3, wherein said polypeptide does not comprise one or more of the domains selected from the group consisting of CCP1, CCP2, CCP3, CCP4, CCP5, CCP7 and CCP8 of the C4BP alpha chain.

5. The oligomer for use according to any one of claims 1 to 4, wherein the polypeptide comprises a full-length C4BP alpha chain.

6. The oligomer for use according to any one of claims 1 to 5, wherein the CCP6 domain of the C4BP alpha chain is SEQ ID NO: 1.

7. The oligomer for use according to any one of claims 1 to 6, wherein the oligomerization domain is SEQ ID NO: 6 or a functionally equivalent variant thereof, preferably SEQ ID NO: 6.

8. The oligomer for use according to any one of claims 1 to 7, wherein the polypeptide consists of SEQ ID NO: 7 or a functionally equivalent variant thereof, preferably SEQ ID NO: 7.

9. The oligomer for use according to any one of claims 1 to 8, wherein the polypeptide consists of a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9 and a functionally equivalent variant thereof or wherein the polypeptide consists of SEQ ID NO: 10 or a functionally equivalent variant thereof, preferably SEQ ID NO: 10.

10. The oligomer for use according to any one of claims 1, 3, 5 or 6, wherein the polypeptide consists of SEQ ID NO: 11 or a functionally equivalent variant thereof.

11. The oligomer for use according to any one of claims 1, 3, 5 or 6, wherein the oligomer is a C4BP isoform selected from the group consisting of a7p0 and a6p0.

12. The oligomer for use according to any one of claims 1 to 11, wherein the disease involving an undesired activation of the immune system is selected from the group consisting of immunoinflammatory disease, sepsis, autoimmune disease, transplant rejection, graft-versus-host disease and hypersensitivity disease, autoinflammatory diseases, liver diseases, metabolic diseases, cardiovascular diseases, cancer, metastasic cancer, renal diseases, infectious diseases, central nervous system diseases, bone diseases, eye diseases, fetal / neonatal alloimmune thrombocytopenia (FNAIT), skin scarring, and oxidized phospholipid-driven lung injury.

13. The oligomer for use according to claim 12, wherein the metabolic disease is selected from the group consisting of hepatic steatosis, metabolic syndrome, obesity, diabetes, hyperlipidemia, atherosclerosis, nephropathy, retinopathy, peripheral neuropathy, non-alcoholic fatty liver disease (NAFLD); wherein the cardiovascular disease is selected from the group consisting of stroke, heart failure, diabetic cardiomyopathy, vasculitis and venous thromboembolism; wherein the cancer is selected from the group consisting of radiotherapy and / or chemotherapy resistant cancer, cancer resistant to HER2-targeted therapies, oral squamous cell carcinoma (OSCC), myeloid leukemia, monocytic leukemia, breast cancer, ovarian cancer, gastric cancer, colorectal cancer, melanoma, colon adenocarcinoma and lung cancer; whereinthe renal disease is selected from the group consisting of chronic kidney disease and lithogenic-induced gallstones; wherein the infectious disease is selected from the group consisting of COVID- 19, hepatitis B and C, HIV-induced AIDS and malaria; wherein the central nervous system disease is Alzheimer’s disease; wherein the bone disease is selected from the group consisting of osteoporosis and periodontitis; wherein the eye disease is selected from the group consisting of age-related macular degeneration, diabetic retinopathy, glaucoma, intraocular neovascularization, intraocular inflammation and retinal inflammation / degeneration.

14. The oligomer for use according to claim 13, wherein the disease is selected from the group consisting of autoimmune and autoinflammatory diseases, hepatic steatosis and cancer.

15. The oligomer for use according to any one of claims 1 to 14, wherein the compound is to be administered by subcutaneous route.

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