GPR120 agonists for the treatment of inflammatory bowel disease

Abstract

The present invention relates to GPR120 agonists for use in the treatment of inflammatory bowel disease, in particular ulcerative colitis or Crohn's disease.

Description

[Technical field]

[0001] The present invention relates to GPR120 agonists for use in the treatment of inflammatory bowel disease. [Background technology]

[0002] Inflammatory bowel disease (IBD) is considered a group of chronic inflammatory conditions of the digestive tract, the most common of which are ulcerative colitis (UC) and Crohn's disease (CD). In ulcerative colitis, inflammation is localized to the mucosal lining of the large intestine (colon) and rectum, whereas in Crohn's disease, inflammation can be patchy and distributed throughout any part of the digestive tract (including the mouth), although the ileum is the most commonly affected area.

[0003] Both diseases share similar symptoms such as diarrhea, abdominal pain, rectal bleeding, and weight loss. However, despite the similarities between them, there are also some other symptoms that are different. For example, malnutrition is common in Crohn's disease because the affected area is the small intestine, which is responsible for nutrient absorption. On the other hand, ulcerative colitis is associated with bloody stools and rectal bleeding, which are uncommon in Crohn's disease. More than 50% of CD patients suffer from folate and vitamin D deficiency, while more than 50% of UC patients suffer from iron deficiency.

[0004] Inflammatory bowel disease affects both men and women equally. It often begins during adolescence, with approximately 25% of IBD patients being under the age of 20. In some cases, it may occur in adulthood (Journal of Medicine and Life Vol.12, Issue 2, April-June 2019, pp.113-122). The causes of inflammatory bowel disease remain largely unknown, but are caused by highly complex interactions between genetic and environmental factors, dysregulated immune responses, and altered microbiota. However, none of these factors alone is sufficient to cause disease. Clinical, endoscopic, histological, and radiological tests are used to diagnose the disease (Zhang Y.et al.World J Gastroenterol 2014;20(1):91-99).

[0005] Currently, the treatment of inflammatory bowel disease is based on a step-by-step approach. In mild to moderate Crohn's disease, the first step in treatment is the use of aminosalicylates, such as 5-aminosalicylic acid (5-ASA) (brand name Mesalamine). If this is ineffective, antibiotics, corticosteroids and immunosuppressants are used as a last resort before hospitalization.

[0006] For ulcerative colitis, conventional treatments include 5-aminosalicylic acid, corticosteroids, and purine analogues (azathioprine and mercaptopurine). If these fail, patients are treated with immunosuppressants such as calcineurin inhibitors, tacrolimus, and TNF-α inhibitors (Baumgart DC et al. The Lancet 2007;369(9573):1641-57).

[0007] However, existing treatment strategies do not always result in remission and can be associated with numerous side effects. Thus, for a significant number of patients, this debilitating disease remains far from being satisfactorily controlled. IBD therefore remains an unmet medical need for many patients (Gordon et al, Eur. J. Gastroenterol. Hepatol. 2015(27), 804-812).

[0008] Therefore, there is a strong need to develop new therapeutic approaches for the treatment of inflammatory bowel diseases that are highly effective and have few side effects. Several studies have shown the beneficial effects of omega-3 polyunsaturated fatty acids administered as adjunctive therapy in the prevention or treatment of ulcerative colitis and Crohn's disease. Omega-3 fatty acids have also been shown to be substrates for the production of protectins, resolvins, and maresins that can modulate and attenuate inflammatory processes, resulting in remission of IBD, and therefore can be considered a new complementary approach to the treatment of these inflammatory conditions (Marton et al, International Journal of Molecular Science 2019, 20(19), 4851).

[0009] GPR120 is a member of the rhodopsin family of G protein-coupled receptors (GPRs) and has been shown to mediate some of the anti-inflammatory and insulin-sensitizing effects of omega-3 fatty acids (Young Oh, Cell. 2010 September 3;142(5):687-698).

[0010] GPR120 has been shown to be abundantly expressed in human intestinal endocrine L, K, or I cells, where it acts as an exo-receptor for free fatty acids. These cells reside in the intestinal villi and face the intestinal lumen in contact with food (Furness, JB et al. Nat. Rev. Gastroenterol. Hepatol. 2013, 10, 729-740).

[0011] Other studies have confirmed that in the terminal ileum and proximal colon, GPR120 is expressed not only by endocrine cells but also by all epithelial cells lining the villus (Paulsen et al, PLoS ONE 2014, 9(2):e88227, doi:10.1371 / journal.pone.0088227). [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] Journal of Medicine and Life Vol. 12, Issue 2, April-June 2019, pp. 113-122 [Non-Patent Document 2] Zhang Y. et al. World J Gastroenterol 2014; 20(1): 91-99 [Non-Patent Document 3] Baumgart DC et al. The Lancet 2007; 369(9573):1641-57 [Non-Patent Document 4] Gordon et al, Eur. J. Gastroenterol. Hepatol. 2015 (27), 804-812 [Non-Patent Document 5] Marton et al, International Journal of Molecular Science 2019, 20 (19), 4851 [Non-Patent Document 6] Young Oh, Cell. 2010 September 3; 142(5): 687-698 [Non-Patent Document 7] Furness, JB et al. Nat. Rev. Gastroenterol. Hepatol. 2013, 10, 729-740 [Non-Patent Document 8] Paulsen et al, PLoS ONE 2014, 9(2): e88227, doi:10.1371 / journal.pone.0088227 Summary of the Invention

[0013] As shown in the experimental section, the inventors have surprisingly identified several structurally related GPR120 agonists that are particularly effective in the prevention or treatment of inflammatory bowel disease. When orally administered, these compounds are poorly absorbed and are highly concentrated in the ileum and colon, allowing them to act locally and effectively in the prevention or treatment of inflammatory bowel disease.

[0014] Furthermore, the inventors have discovered that, unlike other known GPR120 agonists, these compounds do not cause internalization (intercellular transport) and degradation of GPR120 after binding to and activating the receptor, and are therefore able to exert and maintain potent agonist activity.

[0015] Accordingly, a first object of the present invention is to provide a compound of formula (I):

[0016] [ka]

[0017] [In the formula, R 1 is CH3, R 2 is H, R 3 is selected from F, Cl and CF3, or R 1 and R 2 are independently selected from substituted or unsubstituted phenyl or thiophene and H, with the proviso that R 1 or R 2 at least one of is H, and R 3 is F, wherein, if said phenyl or thiophene is substituted, the substituent is preferably selected from Cl, F or CH3, more preferably at the 2-, 3- or 4-position of the phenyl, or the 3-, 4- or 5-position of the thiophene. It is a GPR120 agonist.

[0018] A second object of the present invention is a pharmaceutical composition comprising a GPR120 agonist of formula (I) as defined above, for use in the prevention or treatment of inflammatory bowel disease in an individual. A third object of the present invention is a method for the prevention or treatment of inflammatory bowel disease, said method comprising administering to an individual in need thereof a therapeutically effective amount of a GPR120 agonist of formula (I) as defined above. [Brief description of the drawings]

[0019] [Figure 1] Figure 1 shows the % weight loss over time (Figure 1A) and DAI scores (Figure 1B) in the experimental groups described in Example 1a). DSS I, II, and III indicate the stage of disease progression (acute / chronic). The experiment is described in Example 1b). [Diagram 2]FIG. 2 shows the colon length measured after sacrificing the mice on day 39 in the experimental group described in example 1a), as described in example 1c). [Diagram 3] FIG. 3 shows images of the endoscopic assay described in Example 1d) for each of the experimental groups described in Example 1a). [Figure 4] FIG. 4 shows the composite endoscopic score as described in Example 1d) for each of the experimental groups as described in Example 1a). [Diagram 5] FIG. 5 shows the colon thickening score (FIG. 5A) and vascular pattern score (FIG. 5B) as described in Example 1d) for each of the experimental groups described in Example 1a). [Figure 6] FIG. 6 shows the scores for visible fibrin (FIG. 6A) and mucosal surface granularity (FIG. 6B) as described in Example 1d) for each of the experimental groups described in Example 1a). [Figure 7] Figure 7 shows the histological analysis described in Example 1e) for each of the experimental groups described in Example 1a). Histological sections stained with hematoxylin and eosin are shown. [Figure 8] FIG. 8 shows the mean histological scores for each parameter of the Rachmilewitz score shown in the table of Example 1e) for each of the experimental groups described in Example 1a). [Figure 9] FIG. 9 shows a schematic diagram of GPR120 signal transduction. [Figure 10] Figure 10 shows the results of Western blotting of GPR120 and β-arrestin2 for each of the experimental groups described in Example 1a) (Figure 10B), the results of densitometric analysis of the Western blotting blots performed using Image J, and the ratio between the levels of co-immunoprecipitated β-arrestin2 and GPR120 obtained for each experimental group as described in Example 1f) (Figure 10A). [Figure 11]Figure 11 shows the results of Western blotting of GPR40 and β-arrestin2 (Figure 11B), densitometric analysis of the Western blotting blots performed using Image J, and the ratio between the levels of co-immunoprecipitated β-arrestin2 and GPR120 obtained for each experimental group as described in Example 1f) (Figure 11A). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] definition According to the present invention, the term "prevention" refers to administration to an individual before the establishment or occurrence of a disorder or pathological event, in order to obtain partial or complete prevention thereof.

[0021] According to one aspect of the invention, the prevention is a complete prevention, in which the disorder or pathological event is completely averted. According to an alternative aspect of the invention, prevention is partial prevention, delaying the onset or progression or reducing the severity of a disorder or pathological event.

[0022] According to the present invention, the term "treatment" refers to completely reversing or reducing the severity or progression of a disorder or pathological event after it has been established or has occurred. According to the present invention, the term "individual" refers to a human or an animal, preferably a human.

[0023] According to the present invention, the term "GP120" refers to G protein-coupled receptor 120 (GPR120), also known as free fatty acid receptor 4. According to the present invention, the term "GPR120 agonist" refers to a compound that binds to GPR120 and activates the GPR120 signaling pathway.

[0024] According to the present invention, the term "individual" refers to a human or an animal, preferably a human. Detailed Description of the Invention A first object of the present invention is to provide a compound of formula (I):

[0025] [ka]

[0026] [In the formula, R 1 is CH3, R 2 is H, R 3 is selected from F, Cl and CF3, or R 1 and R 2 are independently selected from substituted or unsubstituted phenyl or thiophene and H, with the proviso that R 1 or R 2 at least one of is H, and R 3 is F, wherein, if said phenyl or thiophene is substituted, the substituent is preferably selected from Cl, F or CH3, more preferably at the 2-, 3- or 4-position of the phenyl, or the 3-, 4- or 5-position of the thiophene. It is a GPR120 agonist.

[0027] Preferably, the GPR120 agonist of formula (I) is 7-(3-(N-(4-fluoro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(4-chloro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(4-trifluoromethyl-2,6-dimethyl-phenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(6-fluoro-4-methyl-[1,1'-biphenyl]-3-yl)sulfamoyl)phenyl)heptanoic acid, 7-(3-{[4-fluoro-2-methyl-5-(thiophen-2-yl)phenyl]sulfamoyl}phenyl)heptanoic acid, and 7-(3-(N-(5-fluoro-3-methyl-[1,1'-biphenyl]-2-yl)sulfamoyl)phenyl)heptanoic acid Selected from.

[0028] A particularly suitable GPR120 agonist of formula (I) for use in accordance with the present invention is 7-(3-(N-(4-fluoro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid, also known as DFL23806.

[0029] A further object of the present invention are novel GPR120 agonists of formula (I) 7-(3-{[4-fluoro-2-methyl-5-(thiophen-2-yl)phenyl]sulfamoyl}phenyl)heptanoic acid.

[0030] As shown in the experimental section, the present inventors have found that the above GPR120 agonists of formula (I) are highly effective in the prevention and treatment of inflammatory bowel disease. Preferably, the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

[0031] Administration of a GPR120 agonist to an individual for use in accordance with the present invention follows known methods. Preferably, said administration is oral or rectal.

[0032] The present inventors have found that the therapeutic effect of GPR120 agonists on inflammatory bowel disease is highly dependent on the local activity of the molecule at the site of inflammation in the digestive tract. Therefore, when GPR120 agonists are orally administered, the delivery of a sufficient amount of active drug to the affected area is essential for the therapeutic effect. At the same time, the systemic use of the compound is not useful for the therapeutic effect, so it should be minimized to improve the therapeutic effect and avoid any systemic side effects.

[0033] The inventors have also found that the GPR120 agonists of formula (I) according to the invention have pharmacokinetic and stability characteristics that make them particularly suitable for administration by oral administration. In particular, as shown in Example 2, the inventors have confirmed that the GPR120 agonists of formula (I), when administered orally, exhibit a pharmacokinetic profile characterized by very low systemic absorption and high concentrations reaching the lower gastrointestinal tract, and are therefore suitable for exerting a local effect at the level of the region of the gastrointestinal tract affected by IBD.

[0034] Moreover, these GPR120 agonists have been found to be highly stable in gastric and intestinal fluids, in intestinal microsomes, and finally to have low permeability to Caco-2 (an immortalized cell line of human colorectal adenocarcinoma cells). Thanks to these characteristics, the GPR120 agonists of formula (I) achieve a local effect at the level of the mucosal layer of the ileum and large intestine, without the need for local delivery by controlled release or gastric resistant (enteric coated) formulations.

[0035] Thus, preferably, GPR120 agonists of formula (I) for use according to the present invention are administered orally. Preferably, the GPR120 agonists of formula (I) for use according to the present invention are administered in the form of a pharmaceutical formulation.

[0036] Preferably, the GPR120 agonists of formula (I) for use according to the present invention are administered in a pharmaceutical formulation which is not a controlled release formulation. Preferably, the GPR120 agonists of formula (I) for use according to the present invention are administered in a non-gastric resistant pharmaceutical composition.

[0037] A further object of the present invention is a pharmaceutical composition comprising a GPR120 agonist of formula (I) as defined above and at least one pharma- ceutically acceptable excipient, for use in the prevention or treatment of inflammatory bowel disease in an individual as described above.

[0038] Preferably, the pharmaceutical composition of the present invention is prepared in a suitable dosage form containing an effective amount of the GPR120 agonist of formula (I) above and at least one pharma- ceutically acceptable excipient. Preferably, the pharmaceutical compositions of the invention are suitable for oral or rectal administration.

[0039] Preferably, the pharmaceutical composition for rectal administration is in the form of a suppository, enema, gel, or foam. Preferably, the pharmaceutical composition for oral administration is in the form of granules, fibres, microparticles, tablets or capsules.

[0040] Preferably, said pharmaceutical composition for oral administration is not a controlled release formulation and / or a gastroresistant formulation. In this application, the term "effective amount" refers to the administration of a compound or composition sufficient to significantly effect a desired clinical response.

[0041] The dosage and treatment regimen of a GPR120 agonist of formula (I) for use in accordance with the present invention for any particular individual will vary depending on a number of factors within the knowledge and expertise of those of ordinary skill in the art, such as the half-life of the particular GPR120 agonist used, the formulation and route of administration used, the individual's age, weight, general health, sex, and diet.

[0042] As described herein, the pharmaceutical compositions of the present invention comprise a GPR120 agonist of formula (I) together with at least one pharma- ceutically acceptable excipient, which excipients, as used herein, are selected from solvents, diluents or other vehicles, dispersing or suspending aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, as appropriate for the particular dosage form desired.

[0043] Some examples of pharma- ceutically acceptable excipients that may be present in the compositions according to the invention include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminium hydroxide; alginic acid; pyrogen-free water; isotonic saline; sterile water; Ringer's solution; buffered saline; dextrose solutions; maltodextrin solutions; ethyl alcohol; and phosphate buffers.

[0044] The dosage forms of the pharmaceutical composition of the present invention can be prepared by techniques well known to pharmaceutical chemists, including mixing, granulating, compressing, dissolving, sterilizing, and the like. Furthermore, the compositions of the present invention may be suitably formulated using any suitable method known in the art or by the methods disclosed in Remington's Pharmaceutical Sciences (latest edition), Mack Publishing Company, Easton, Pa.

[0045] The dosage forms may also contain other conventional ingredients, such as preservatives, stabilizers, surfactants, buffers, osmotic regulators, emulsifiers, sweeteners, colorants, flavoring agents, and the like. A further aspect of the invention is a method for the prevention or treatment of inflammatory bowel disease in an individual, which method comprises administering to said individual an effective amount of a GPR120 agonist of formula (I) as defined above. EXAMPLES

[0046] Experimental section statistical analysis In all experiments, statistical analysis was performed using GraphPad Prism 7 (GraphPad Software). Data are presented as mean ± SD or ± SEM, and differences were considered statistically significant when P < 0.05. ANOVA multivariate analysis with post hoc correction tests was performed for all experiments.

[0047] Example 1 - Therapeutic effect of 7-(3-(N-(4-fluoro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid (DFL23806) in a mouse model of dextran sulfate sodium-induced colitis a) Model setup and treatment protocol The therapeutic effect of oral administration of DFL23806 on colitis was studied using a mouse model in which colitis was induced by administration of dextran sodium sulfate (DSS) (Wirtz S. et al. Nature Protocols; Volume: 2, Issue: 3 Pages: 541-546; 2007).

[0048] Specifically, 8- to 12-week-old female C57 / Bl6 mice were given three oral cycles of 2.5% dextran sodium sulfate (DSS) (molecular weight 40 kDa; MP Biomedicals) in drinking water, with each cycle featuring 7 days of DSS administration followed by 7 days of filtered purified water.

[0049] DSS administration induces clinical signs of disease as early as one day after treatment, resulting in altered expression of tight junction proteins and increased expression of inflammatory cytokines. This initial minor effect is followed by increasingly dramatic symptoms including increased intestinal permeability, goblet cell loss, epithelial erosions and ulcers, and severe bleeding, as described below (which is even more pronounced in chronic models). More specifically, the current DSS-induced colitis model is characterized by an acute and a chronic phase, with acute clinical symptoms (diarrhea and / or gross bloody stools) associated with the presence of erosions and inflammation. More importantly, the earliest histological changes are represented by the loss of one-third of the crypt base (3 days after the first DSS administration), progressing over time to the loss of the entire crypt, resulting in erosions by day 5. The earliest changes are very localized and are not associated with inflammation, which is a secondary phenomenon and only becomes evident after the appearance of erosions.

[0050] Animals treated with multiple cycles of DSS followed by water cycles develop chronic colitis characterized by the following histological features: areas of erosion and inflammation, hypoactivity, crypt distortion, reddish epithelial proliferation and possible dysplasia. Clinical disease activity indices correlate significantly with pathological changes in both acute and chronic phases of the disease (Chassaing, B, Current Protocols in Immunology 2014, 104(1), Cooper, HS et al, Laboratory Investigation; a Journal of Technical Methods and Pathology 1993, 69(2), 238-249, D'Alessio, S et al, Journal of Clinical Investigation 2014, 124(9), 3863-3878).

[0051] From the third day onwards after DSS administration, the mice were divided into four experimental groups and received the treatments specified below. - Group 1: DSS (n=8) No treatment was used as a negative control.

[0052] - Group 2: DSS + vehicle (n = 8) Treatment with 20% v / v dimethyl sulfoxide (DMSO) and 80% v / v phosphate-buffered saline (PBS), 0.1 M, pH 8 (vehicle), served as negative control.

[0053] - Group 3: DSS+DFL 90mg / kg (n=8) Treatment with DFL23806 dissolved in vehicle, dose 90 mg / kg once daily. - Group 4: DSS+5-ASA 60mg / kg (n=8) Treatment with 5-aminosalicylic acid (5-ASA) in vehicle at a dose of 60 mg / kg once daily was used as the reference positive control.

[0054] Vehicle, DFL23806 and 5-ASA were administered every morning by oral gavage using a plastic sterile feeding tube 20ga x 30mm. All mice were sacrificed on day 39.

[0055] b) Evaluation of weight loss and DAI (Disease Activity Index) score The DAI score was established based on a scale that takes into account various parameters characterizing the induction and progression of experimental colitis (RS Walmsley et al. Gut 1998;43:29-32).

[0056] Body weight, the presence of visible blood in the stool, and stool consistency were recorded every 2 days. In particular, DAI was determined at each time point by scoring the change in body weight loss (0=none, 1=1%-5%, 2=5%-10%, 3=10%-20%, 4=>20%), stool consistency (0=normal, 2=loose stool, 4=diarrhea), and rectal bleeding (0=normal, 2=occult blood, 4=gross bleeding).

[0057] The results show that administration of DFL23806 has a significant effect on both the reduction in weight loss (Figure 1A) and the reduction in DAI (Figure 1B) compared to controls (DSS and DSS+vehicle groups) already at day 7. Interestingly, DFL23806 was found to be even more effective than 5-ASA at certain time points.

[0058] c) Measurement of colon length Mice were sacrificed on day 39, colons were harvested and colon length was measured for each mouse as an additional parameter of intestinal inflammation.

[0059] The results confirmed that DFL23806 and 5-ASA showed significant inhibitory effects as measured by colon length compared to the DSS+vehicle group (Figure 2). d) Endoscopic evaluation and score On the day of sacrifice (day 39), mice were anesthetized with 2% isoflurane and subjected to endoscopic examination to confirm the degree of inflammation. The experimental endoscopic set-up, called the “Coloview system”, consisted of a miniature endoscope (1.9 mm outer diameter endoscope), a xenon light source, a triple-tip camera, and an air pump to achieve controlled distension of the mouse colon (all manufactured by Karl Storz, Tuttlingen, Germany). The endoscopic procedure was observed on a color monitor and a Modified Mouse Endoscopic Index score of colitis severity was assigned based on colonic translucency (0-3 points), granular character of the mucosa (0-3 points), morphology of the vascular pattern (0-4 points), and presence of fibrin (0-4 points) as previously described by Dr. D'Alessio (D'Alessio, S. et al. J. Clin. Invest. 124, 3863-3878; 2014), giving a cumulative score ranging from 0 (no signs of inflammation) to 16 (very severe signs of inflammation by endoscopy).

[0060] The composite endoscopic scores obtained showed that DFL23806 and 5-ASA could significantly inhibit experimental chronic colitis comparable to the controls (DSS and DSS+vehicle groups) (Figures 3 and 4). This was observed not only in terms of the composite endoscopic score (Figure 4), but also in terms of colon thickness (Figure 5A), vascular pattern (Figure 5B), visible fibrin (Figure 6A), and granularity of the mucosal surface (Figure 6B). Increased mucosal granularity represents edema and small erosions. Edema of the lamina propria is characterized by clusters of neutrophils and capillaries. These data indicate that promoting the resolution of inflammation through oral administration of a selective GPR120 agonist such as DFL23806 may help reduce neutrophil infiltration and mucosal edema.

[0061] e) Histological evaluation and scoring Histological analysis was performed blindly by an expert pathologist, and intestinal inflammation was graded. More specifically, colons of colitic mice from the various experimental groups were evaluated for histological analysis using 2 μM paraffin-embedded sections stained with hematoxylin (Dako) and eosin (Diapath). A blinded pathologist assessed the degree of inflammatory cell infiltration and mucosal damage using the RACHMILEWITZ score (Table 1) (Rachmilewitz, D. et al. Gastroenterology 122, 1428-1441; 2002).

[0062] This scoring system does not distinguish between proximal and distal colon and takes into account five histological parameters, namely ulceration, degree of ulceration, flogosis, degree of inflammation, and fibrosis in the entire colonic section, as shown in Table 1. For each parameter, a score of 0 to 4 is given, as shown in Table 1. In this example, fibrosis was not evaluated in the DSS-induced chronic colitis model (Rachmilewitz, D. et al. Gastroenterology 122, 1428-1441; 2002), since this parameter does not develop fibrosis.

[0063] Table 1. Rachmilewitz scores

[0064] [Table 1]

[0065] Representative histological images in Figure 7 show deep ulcers with crypt loss and extensive inflammatory infiltrates in the DSS and DSS+vehicle groups. Furthermore, the results confirmed that both DFL and 5-ASA significantly reduced ulcers, ulcer extent, inflammation, and histological scores of inflammation extent when compared to DSS+vehicle (Figure 8).

[0066] f) Assessment of binding of DFL to GPR120 and GPR40 The efficiency of DFL in activating the GPR120 and / or GPR40 receptors was examined. Agonist-induced GPR120 can engage multiple signaling pathways to regulate overt physiological outcomes. After ligand binding, β-arrestins, such as β-arrestin2, can associate with the cytoplasmic domain of GPR120 and link the receptor to specific downstream signaling pathways (Figure 9). From this, we verified the efficiency of agonists in activating GPR120 and GPR40 by immunoprecipitation. Colon lysates (~900 μg protein) from six mice in each experimental group were pooled and precleared with Protein A / G plus agarose (Santa Cruz), then incubated with the antibodies used for immunoprecipitation (anti-GPR120 or anti-GPR40, Abcam) for 16 h at 4 °C, and then incubated with Protein A / G plus agarose for 5 h. The agarose beads were then collected by centrifugation, washed four times with lysis buffer, and heated at 95°C for 5 min after adding Laemmli buffer. The resulting immunoprecipitates were resolved by SDS-PAGE and probed with anti-β-arrestin 2 antibody (Santa Cruz).

[0067] After Western blotting, densitometric analysis of the blots was performed using Image J. Figures 10 and 11 show the ratio between the levels of coimmunoprecipitated β-arrestin2 and GPR120 (Figure 10) or GPR40 (Figure 11) and the blotting results.

[0068] The results show that DFL efficiently activates GPR120 on day 39. Indeed, the association between GPR120 and β-arrestin2 in the DSS+DFL group is higher than that in the control group or the DSS+5-ASA 60 mg / kg group (Figure 10).

[0069] By immunoprecipitation with anti-GPR40 antibody, the results showed that there was no increase in the association of GPR40 with β-arrestin2 in the DSS+DFL or DSS+5-ASA groups compared with mice treated with DSS or DSS+vehicle alone (Figure 11).

[0070] This confirmed that DFL23806 is a selective agonist of GPR120 and can mediate signaling through the β-arrestin pathway. Example 2 – Pharmacokinetic evaluation of GPR120 agonists after oral administration The objective of this study was to evaluate the plasma and tissue exposure of the following compounds following a single oral dose in mice: 7-(3-(N-(4-fluoro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid (hereinafter referred to as DFL23806), 7-(3-(N-(4-chloro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid (hereinafter referred to as DFL23914) 7-(3-(N-(2,6-dimethyl-4-(trifluoromethyl)phenyl)sulfamoyl)phenyl)heptanoic acid (hereinafter referred to as DFL23922) 7-(3-(N-(6-fluoro-4-methyl-[1,1'-biphenyl]-3-yl)sulfamoyl)phenyl)heptanoic acid (hereinafter referred to as DFL23916) 7-(3-(N-(5-fluoro-3-methyl-[1,1'-biphenyl]-2-yl)sulfamoyl)phenyl)heptanoic acid (hereinafter referred to as DFL23917) 7-(3-{[4-fluoro-2-methyl-5-(thiophen-2-yl)phenyl]sulfamoyl}phenyl)heptanoic acid (hereinafter referred to as DFL24102) For each of the above compounds, nine male CD1 mice, approximately 6 / 7 weeks of age at the start of the treatment period, were orally treated by gavage with 55 mg / kg of test compound (purity adjusted) dissolved in a vehicle consisting of 20% v / v dimethylsulfoxide (DMSO) and 80% v / v phosphate buffered saline (PBS), 0.1 M, pH 8.

[0071] For pharmacokinetic evaluation in plasma, multiple retro-orbital blood sampling was performed pre-dose or 30 min, 1, 2, 6, 8 and 24 h post-dose (3 mice / time point). Blood samples were collected in heparinized blood collection tubes. Samples were immediately placed on ice and kept cold until centrifugation (10000g, approx. +4°C for 3 min). Approximately 50 μL of plasma was stored in a -80°C freezer until analysis.

[0072] All animal procedures (e.g. housing, health care, restraint, dosing) and ethical review were performed in accordance with current Italian legislation (Legislation, March 4, 2014, no. 26) implementing Directive 2010 / 63 / UE on the protection of animals used in biomedical research.

[0073] After sample collection, bioanalysis of test compound concentrations was performed using an HPLC-MS / MS method, and pharmacokinetic analysis of compounds in plasma and tissues was performed according to standard noncompartmental methods using the Watson system (v 7.6, Thermo Fisher Scientific, Waltham, MA, USA) on mean concentration data.

[0074] The pharmacokinetic parameters obtained for each compound tested are summarized in the table below. Table 2. Pharmacokinetic parameters after oral administration in mice.

[0075] [Table 2]

[0076] The biodistribution of DFL23806 in the proximal and distal colon was evaluated following a single oral dose of 90 mg / kg DFL23806 in male Swiss albino mice. The results obtained are summarized in the table below.

[0077] Colonic concentrations (ng / g) of DFL23806 following PO administration (90.0 mg / kg) to male Swiss albino mice.

[0078] [Table 3]

[0079] Colon / plasma ratio of DFL23806 following PO (90.0 mg / kg) administration to male Swiss albino mice

[0080] [Table 4]

[0081] As can be seen, the concentration of DFL23806 in the colon was higher than the corresponding concentration measured in plasma (ratio=64.2). These results indicate that the tested compounds are concentrated in the colon and therefore exert their activity locally.

[0082] Example 3 – Internalization of GPR120 after activation We have assessed the internalization of human GPR120 after activation with compounds DFL23806, DFL23916, DFL23914, DFL23922, DFL23917 and DFL24102 using the Enzyme Fragment Complementation (EFC) technique with β-galactosidase developed by DiscoverX in dose-response curves up to 100 μM. The results show that for all compounds tested, the concentration of GPR120 agonist required to activate the receptor is significantly lower than the concentration that results in GPR120 internalization and subsequent degradation. In particular, DFL23806 has an AC50 of 3.9 μM and activates the internalization of human GPR120 receptor with 59.1% efficacy at 100 μM.

[0083] The large difference between the AC50 values ​​for activation and internalization suggests that the receptor is not internalized immediately after compound binding, and therefore treatment with a GPR120 agonist according to the present invention does not result in downregulation of the receptor.

[0084] This feature is not observed in other GPR120 agonists known in the prior art. For example, data on the molecules TUG-891 and GW9508 show AC activation and internalization. 50 It is clear that the values ​​are similar (0.065 μM and 0.051 μM, respectively, for TUG-891 and 6.8 μM and 9.6 μM, respectively, for GW9508).

[0085] These concentrations suggest that there is rapid internalization and turnover of the receptor following binding of either of these compounds. The delayed internalization of the receptor mediated by the GPR120 agonists of the present invention increases the potency of action since activated GPR120 can participate in signal transduction for a longer period of time.

Claims

1. 1. A pharmaceutical composition for use in the prevention or treatment of inflammatory bowel disease in an individual, comprising a compound of formula (I): 【Chemistry 1】 [In the formula, R １ is CH ３ and R ２ is H, and R ３ is F, Cl and CF ３ Choose from or R １ and R ２ are independently selected from substituted or unsubstituted phenyl or thiophene and H, with the proviso that R １ or R ２ is H, and R ３ is F, wherein, if said phenyl or thiophene is substituted, the substituent is preferably Cl, F or CH ３ and more preferably at the 2-, 3- or 4-position of the phenyl, or the 3-, 4- or 5-position of the thiophene. A pharmaceutical composition comprising a GPR120 agonist of the formula:

2. A GPR120 agonist, 7-(3-(N-(4-fluoro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(4-chloro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(4-trifluoromethyl-2,6-dimethyl-phenyl)sulfamoyl)phenyl)heptanoic acid, 7-(3-(N-(6-fluoro-4-methyl-[1,1′-biphenyl]-3-yl)sulfamoyl)phenyl)heptanoic acid, 7-(3-{[4-fluoro-2-methyl-5-(thiophen-2-yl)phenyl]sulfamoyl}phenyl)heptanoic acid, and 7-(3-(N-(5-fluoro-3-methyl-[1,1'-biphenyl]-2-yl)sulfamoyl)phenyl)heptanoic acid The pharmaceutical composition according to claim 1, selected from:

3. The pharmaceutical composition of claim 1, wherein the GPR120 agonist is 7-(3-(N-(4-chloro-2,6-dimethylphenyl)sulfamoyl)phenyl)heptanoic acid.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.

5. The pharmaceutical composition according to any one of claims 1 to 3, wherein the GPR120 agonist is administered orally.

6. A pharmaceutical composition described in any one of claims 1 to 3, which is not a controlled release formulation and / or a gastric resistant formulation.

7. A pharmaceutical composition described in any one of claims 1 to 3, further comprising at least one inactive pharmaceutically acceptable excipient.

8. The pharmaceutical composition according to claim 7, for oral administration.

9. A GPR120 agonist that is 7-(3-{[4-fluoro-2-methyl-5-(thiophen-2-yl)phenyl]sulfamoyl}phenyl)heptanoic acid.

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