Guanidine alpha-aminoacyl amide derivative compounds, pharmaceutical composition comprising said compounds, methods of synthesis and use thereof

Abstract

A compound derived from guanidine alpha-aminoacyl amide of general formula (I); pharmaceutical compositions comprising said compounds, methods of treatment and methods of synthesis of said compounds.

Description

TITLEGuanidine alpha-aminoacyl amide derivative compounds, pharmaceutical composition comprising said compounds, methods of synthesis and use thereof.TECHNICAL FIELD OF THE INVENTION

[0001] The present invention relates to a novel family of chemical compounds derived from guanidine alpha-aminoacyl amides which are inhibitors of members of the Rho family of GTPases, in particular RAC1 protein. The present invention also relates to compositions, uses and methods of synthesis thereof.BACKGROUND OF THE INVENTION

[0002] Acute and chronic hepatic diseases are a growing cause of morbidity and mortality worldwide, leading to over two million yearly deaths.

[0003] Acute liver failure (ALF) is a critical medical condition characterized by a rapid deterioration of hepatic function in the absence of pre-existing liver disease [1], Massive hepatic necrosis frequently leads to multi-organ failure and is associated with high mortality rates [1, 2], A wide variety of factors can trigger ALF; hepatic injury induced by drugs, such as acetaminophen (APAP), acute viral hepatitis, toxins and auto-immune diseases are the most frequent ALF causes in developed countries. APAP-induced liver injury is initially caused by mitochondrial oxidative stress and dysfunction resulting from the toxic metabolite NAPQI [3].

[0004] RAC1 protein (Ras-related C3 botulinum toxin substrate 1) is a small GTPase involved in cytoskeleton reorganization, ROS migration, proliferation and production, and pro-inflammatory cytokine release [8] [9]

[0010]

[0011] , Notably, some of these processes are involved in ALF, liver cirrhosis and hepatocellular carcinoma (HCC) pathogenesis. Accordingly, RAC1 is involved in NADPH oxidase activation, producing oxygen reactive species that induce hepatocellular damage and exacerbate inflammation

[0011]

[0012] , Likewise, RAC1 has also been linked to a pro- inflammatory macrophage phenotype and to the crucial NLRP3 inflammasome protein complex activation in the development of ALF and chronic liver disease. Furthermore, RAC1 activation promotes the transition of quiescent hepatic stellatecells to a pro-fibrotic active state that releases type I collagen. Finally, it has been shown that RAC1 triggers a transcriptional program related to the distinctive features of HCC aggressiveness.

[0005] As a consequence of damage, necrotic hepatocytes release a large amount of molecules known as damage-associated molecular patterns (DAMPs), resulting in a second wave of hepatocyte death caused by increased sterile inflammation [4]. In addition, in ALF secondary to viral and autoimmune hepatitis, the liver parenchyma is directly damaged by excessive release of inflammatory mediators (such as ROS and cytokines) by resident and recruited immune cells [2, 5], In both scenarios, reactive oxygen species (ROS) overproduction and immune system hyperactivation finally lead to massive hepatocyte necrosis and apoptosis [2], N-acetylcysteine (NAC) is the standard treatment for APAP-related ALF, and is currently also under research for non-APAP-related ALF patients [6]. NAC presents a limited therapeutic time window and may be ineffective in patients with high APAP overdoses [3], Although improvements in ALF medical management have reduced mortality, liver transplant continues to be practiced in about 30% of patients [7, 8]. As a result, there is an urgent need for new therapeutic options to improve the outcomes of these conditions.

[0006] RAC1 is a member of the RHO subfamily of GTPases, which in turn belongs to the RAS superfamily of GTP-binding proteins [9], The change from GDP-bound inactive state to the GTP-bound active state in RAC1 is strictly regulated by several guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs), while their function is modulated by a wide variety of downstream effectors

[0010] , This relates RAC1 to various cellular processes, such as cellular migration, invasion, cell cycle progression and proliferation [10-12], Moreover, it has been widely reported that RAC1 regulates the intracellular production of reactive oxygen species (ROS) derived from NADPH oxidase (NOX) in several cell types [13, 14], For example, sustained RAC1 -dependent NOX activation in hepatic stellate cells results in an excessive ROS production in a mouse model of fibrosis and promotes liver damage

[0015] , In addition, there is growing evidence showing a tight connection between RAC1 and inflammatory processes, such as cytokine production, inflammasome activation, macrophage modulation, neutrophil migration and activation, and the like [16-25], It has been reported that ablation of RAC1 in the myeloid lineage provides protection against inflammation-mediated kidney damagein mice by suppressing the production of IL-6 and TNFa in macrophages

[0016] . Therefore, it is presumed that RAC1 activation may contribute to the development of oxidative and inflammatory damage in ALF.

[0007] The fundamental role of RAC1 in NADPH oxidase activation in several liver cell types has been widely recognized

[0014] . RAC1 plays a key role in the production of reactive oxygen species (ROS) in hepatocytes [26, 27] and immune cells such as monocytes, macrophages and neutrophils [28-31], Through this double effect, RAC1 seems to be crucial for hepatocyte apoptosis induced by oxidative stress, triggered by both intracellular ROS within hepatocytes and ROS secreted by immune cells. In this respect, prior studies have shown an anti-oxidant effect of RAC1 inhibition on several conditions associated with oxidative stress [32-34], but no reports have been produced in the context of ALF.

[0008] In prior reports, the inventors have shown that there is a direct link between RAC1 activation and inflammation and ROS accumulation in the context of ALF. In addition, they have found evidence supporting the notion that RAC1 activation is in fact a relevant feature in ALF, both in humans and in murine models, highlighting the potential of targeting RAC1 as a therapeutic strategy for the management of ALF.

[0009] Acute-on-chronic liver failure (ACLF) is a syndrome characterized by acute decompensation of a chronic liver disease, associated with failure in different organs and a high mortality.

[0010] There are no currently available treatments that specifically target ALF, liver cirrhosis and acute-on-chronic liver failure (ACLF). In this scenario, liver transplant continues to be the only curative option for patients suffering from decompensated disease. Moreover, HOC is a common cause of death among compensated cirrhosis patients, and its incidence and mortality are on the rise. However, most patients are diagnosed at a time when curative therapies are no longer applicable. In addition, the treatments for advanced HCC have not met clinical expectations. Notably, as opposed to other diseases for which knowledge of the molecular mechanisms involved in their cellular pathophysiology has allowed identification of specific molecular targets and the development of “targeted therapies”, the progress in the development of new therapies for ALF, cirrhosis and HCC have been meager.

[0011] In consequence, the RAC1 pathway represents a target candidate for reducing hepatic inflammation, deactivating HSCs and reducing HCC growth.

[0012] In the state of the art there are patent documents related to the development of chemical inhibitors of the RAC1 protein, such as US2024067613, owned by HANMI PHARMACEUTICAL CO LTD; US2016317516, by Univ Indiana Res & Tech Corp, US9278956, by the University of Puerto Rico, US9745257, by Chemo Research SL, and also others that have proposed the use of interference RNA molecules. However, to date, the development of novel inhibitor molecules of RhoGTPase, and in particular RAC1, continues to be an unsolved problem.

[0013] The inventors of the present invention have developed and characterized a novel family of RAC1 inhibitors which are capable of inhibiting the activation of RAC1 by interfering in the interaction between TIAM, a guanine nucleotide exchange factor (GEF), and RAC1.SUMMARY OF THE INVENTION

[0014] An object of the present invention is to provide a compound derived from guanidine alpha-aminoacyl amide of general formula (I):A- CH.or a pharmaceutically acceptable salt thereof, or any of its stereoisomeric forms or a mixture thereof, where A is independently selected from the group consisting of CH and N; R1 is selected from the group consisting of: H; (C1-C4) alkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Br and I; mono- or dihydroxy aryl; 6- membered heteroaryl containing 1 heteroatom selected from the group consisting of N, O, and S; methyl benzoate; methyl acetate; S-methyl imidazole propanoate; R2 is selected from the group consisting of: H (hydrogen), substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Br and I; mono- or dimethoxy phenethyl,benzo[1,3]dioxolyl, N-indolyl methyl; R3 is selected from the group consisting of: H; (C1-C4) alkyl; (C5-C6) cycloalkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Brand I; tert-butyl (disubstituted propyl), mono- or dimethoxy phenethyl, N-indolyl ethyl.

[0015] In a preferred embodiment, the compound derived from guanidine alpha¬ aminoacyl amide of general formula (I) of the present invention comprises substituent A which is selected from the group consisting of CH or N-, R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4-chlorophenyl; 4- fluorophenyl; 3,5-dimethylphenyl; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyl; 2,5-dichlorophenyl; 1-methyl-1H-indol-3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1 H-indol-3-yl)ethyl; 4-bromo-2-methylphenyl; tert-butyl; p-tolyl. In a more preferred embodiment, if A is - CH then R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4- chlorophenyl; 4-fluorophenyl; 3,5-dimethylphenyl; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyl; 2,5-dichlorophenyl; 1- methyl-1H-indol-3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1H- indol-3-yl)ethyl; 4-bromo-2-methylphenyl; tert-butyl; p-tolyl. In a further more preferred embodiment, if A is -N-, then R1 is selected from the group consisting of: methyl; 4-chlorophenyl; R2 is selected from the group consisting of: H, 2,5- dichlorophenyl; R3 is selected from the group consisting of: p-tolyl; 3,4-dimethoxy phenethyl.

[0016] In a yet more preferred embodiment, the compound derived from guanidine alpha-aminoacyl amide of the present invention is selected from the group consisting of:methyl 2-(N-(2-(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamido)acetateN-(2-((4-bromo-2-methylphenyl)amino)-2-oxo-1-phenylethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-(4-hydroxyphenyl)benzamideN-(2-(t-butylamino)-2-oxo-1-phenylethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-(pyridin-3-yl)benzamide- 2-(N-(2-(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamido)acetic acidN-(1 -(benzo[d][1,3]dioxol-5-yl)-2-(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamide- methyl 2-(3-(3-(3,5-dimethylphenyl)guanidino)-N-(2-((4- methoxyphenethyl)amino)-2-oxoethyl)benzamido)benzoatemethyl 2-(N-(1-(benzo[d][1,3]dioxol-5-yl)-2-(cyclohexylamino)-2-oxoethyl)- 3-(3-(3,5-dimethylphenyl)guanidino)benzamido)-3-(1H-imidazol-5- yl)propanoate- N-(2-((2-(1H-indol-3-yl)ethyl)amino)-1-(1-methyl-1H-indol-3-yl)-2- oxoethyl)-N-(3,5-dimethylphenyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamideN-(1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-methylbenzamideN-(1-(2-chlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-methylbenzamideN-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3- (3-(3,5-dimethylphenyl)guanidino)benzamide- N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-(4-fluorophenyl)benzamideN-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-(4-hydroxyphenyl)benzamideN-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3- (3-(4,6-dimethylpyridin-2-yl)guanidino)benzamideN-(1-(2!5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(4,6- dimethylpyridin-2-yi)guanidino)-N-methylbenzamide

[0017] The compound derived from guanidine aipha-aminoacyi amide of the present invention may be used for the treatment of a medical condition mediated by the Rho GTPase cell protein, preferably RAC1. In an embodiment, said medical condition is selected from the group consisting of chronic liver inflammation, acute liver failure, acute-on-chronic liver failure and liver fibrosis. In a further preferred embodiment, said medical condition is selected from the group consisting of liver cancer,pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

[0018] The present invention also provides a pharmaceutical composition comprising a compound derived from guanidine alpha-aminoacyl amide of the present invention and at least one excipient, one adjuvant, ora combination thereof. Said pharmaceutical composition may be used for the treatment of a medical condition characterized by an increase in Rho GTPase cell protein activity, preferably where said Rho GTPase is RAC1. In an embodiment, said medical condition is selected from the group consisting of acute liver failure, chronic liver inflammation, acute-on-chronic liver failure and liver fibrosis. In a further embodiment, said medical condition is selected from the group consisting of liver cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

[0019] The present invention also provides a method for the treatment of a medical condition mediated by the RAC1 cell protein, characterized by comprising the administration to a subject in need thereof of a therapeutically effective amount of at least one compound derived from guanidine alpha-aminoacyl amide of general formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or isomer thereof. In an embodiment, said method for the treatment of a medical condition mediated by the RAC1 cell protein comprises the administration to a subject in need thereof of a therapeutically effective amount of a pharmaceutical composition comprising a compound derived from guanidine alpha-aminoacyl amide of general formula (I). In a preferred embodiment, said treatment is used for the treatment of a medical condition selected from the group consisting of acute liver failure, chronic liver inflammation, acute-on-chronic liver failure and liver fibrosis. In a further preferred embodiment, said treatment comprises treating a medical condition selected from the group consisting of liver cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breastcancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

[0020] A further object of the present invention is to provide a method for the synthesis of a compound derived from guanidine alpha-aminoacyl amide of general formula (I) of the present invention, where said method, if A=CH, comprises the steps of: 1)- obtaining the chemical precursor N-(f-butoxycarbonyl)-N'-(3,5-dimethylphenyi)thiourea (1D-195); 2)- from the precursor of step 1 obtaining the intermediate 3-(2-(t-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid (1 F-123); 3)- obtaining the guanidine alpha-aminoacyl amide compounds according to claims 1 to 5 by Ugi reaction (1 F-181, 1 F-182, 1 F-183, 1 F-184, 1 F-188, 1 F-189, 1 F-190, 1G-54, 1G-55, 1G-56, 1G-57, 1G-58, 1G-59). In a preferred embodiment, said method in said step 1 comprises the following sub-steps: a) preparing a solution of thiourea in dry tetrahydrofuran; b) adding sodium hydride and stirring at room temperature and cooling, preferably to 0 °C; c) adding di-tert-butyl dicarbonate and stirring, preferably at room temperature; d) cooling and adding a second portion of sodium hydride while stirring; e) adding trifluoroacetic anhydride dropwise and stirring, preferably at 0°C; f) adding 3,5-dimethylanillne dropwise, and allowing to reach room temperature and stirring; g) cooling to 0 °C and adding water dropwise to quench the reaction; h) extracting with EtOAc; i) washing the organic layer with brine and drying; j) removing the solvents in vacuo’ k) purifying by flash chromatography (silica gel) (hexane-EtOAc), and I) recrystallizing from MeOH-H2O to provide N-(f-butoxycarbonyl)-N'-(3,5-dimethylphenyl)thiourea as a white solid (1D-195) (35, 36).

[0021] In a preferred embodiment, said step 2) comprises the following sub-steps: a) to the solution of N-(f-butoxycarbonyl)-N'-(3,5-dimethylphenyl)thiourea (1D-195) obtained in said step 1, adding benzyl 3-aminobenzoate and triethylamine in anhydrous CH2Cl2; b) cooling to 0 °C and treating with EDCI; c) stirring under argon, preferably for 1 h; d) stirring at room temperature; e) purifying the residue by flash chromatography (silica gel) (eluted with a gradient of hexane: EtOAc); f) dissolving the product obtained in step e) in EtOAc and carrying out catalytic hydrogenation with 10% palladium on activated charcoal at room temperature; g) filtering the reaction mixture through a Celite bed and evaporating the solvent at reduced pressure, to provide the desired intermediate 3-(2-(t-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid (1F-123) (37).

[0022] In a preferred embodiment, said step 3) comprises the following sub-steps: a) to the stirring solution of 3-(2-(t-butoxycarbonyl)-3-(3,5- dimethylphenyl)guanidino)benzoic acid (1F-123) obtained in step 2, adding an amine and an aldehyde in MeOH; b) adding the corresponding isonitryl; c) stirring, preferably for 2 to 7 days, at room temperature; d) evaporating the solvent and purifying by flash chromatography (silica gel); e) treating the product obtained in step d) with 15.5% HCI in isopropanol, preferably for 4-5 h at room temperature; f) slowly adding NaHCO3 under suitable conditions until reaching pH 8-9; g) extracting with CH2CI2; h) drying the combined organic layers (Na2SO4and evaporating the solvent; i) purifying by column chromatography (silica gel) to provide the compounds derived from guanidine alpha-aminoacyl amide that are the main object of the present invention, in a more preferred embodiment, when said compound derived from guanidine alpha-aminoacyl amide is N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p- tolylamino)ethyl)-3-(3-(3,5-dimethylphenyl)guanidino)-N-methylbenzamide (1G-55), then said sub-step 3 comprises the following steps: a) adding 2,5- dichlorobenzaldehyde, methylamine hydrochloride, triethylamine, 4-methylphenyl isonitrile and 3-(2-(t-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid in 1 mL of MeOH; b) stirring at room temperature, preferably for 6 days; c) purifying the product of step b by column chromatography (silica gel) eluted with hexane- EtOAc; d) treating with HCI in IPA; e) purifying by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p- tolylamino)ethyl)-3-(3-(3,5-dimethylphenyl)guanidino)-N-methylbenzamide.

[0023] A further object of the present invention is to provide a method for the synthesis of a compound derived from guanidine alpha-aminoacyl amide of general formula (I) where if A is N, and said derivative is N-(1-(2,5-dichlorophenyl)-2-oxo-2- (p-tolylamino)ethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)-N-methylbenzamide, then said method comprises the steps of: 1) obtaining the chemical intermediate 3-amino-N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-N-methyl-benzamide from a solution of 3-nitrobenzoic acid, an aldehyde, an amine in MeOH, adding an isonitryi by Ugi reaction followed by catalytic hydrogenation; 2) obtaining the chemical intermediate N-(4,6-dimethylpyridin-2-yl)cyanamide from a solution of N-(4,6-dimethyl-2-pyridyl)thiourea, a KOH solution, and adding a 1,5 M lead acetate solution (Process for the preparation of the compound N-(3,5-dimethylphenyl)~N’(2- trifluoromethylphenyl)guanidine., US20180244608 A1); 3) coupling theintermediates obtained in step 1 and step 2, under suitable conditions, to provide N-(1-(2!5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(4,6-dimethylpyridin-2- yl)guanidino)-N-methylbenzamide.

[0024] A further object of the present invention is to provide a method for the synthesis of a compound derived from guanidine aipha-aminoacyi amide of general formula (I) where if A is N, and said derivative is in particular N-(4-chlorophenyl)-N- (2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(4,6-dimethylpyridin-2- yi)guanidino)benzamide, then said method comprises the following steps: 1) obtaining the chemical intermediate 3-amino-N-(4-chlorophenyl)-N-(2-((3,4- dimethoxyphenethyl)amino)-2-oxoethyl)benzamide from a solution of 3-nitrobenzoic acid, an aldehyde, an amine in MeOH, adding an isonitryl by Ugi reaction followed by catalytic hydrogenation; 2) obtaining the chemical intermediate N-(4,6- dimethylpyridin-2-yl)cyanamide from a solution of N-(4,6-dimethyl-2- pyridyl)thiourea, a KOH solution, and adding a 1,5 M lead acetate solution; 3) coupling the intermediates obtained in step 1 and step 2, under suitable conditions, to provide N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)benzamide

[0025] The present invention also provides a pharmaceutical composition for the treatment of a medical condition mediated by the RAC1 cell protein, where said composition comprises at least one compound derived from guanidine alpha¬ aminoacyl amide of general formula (I), where A is independently selected from the group consisting of CH or N; and where if A is -CH- then R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4-chlorophenyl; 4-fluorophenyl; 3,5-dimethylphenyl; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyl; 2,5-dichlorophenyl; 1-methyl-1H-indol-3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1 H-indol-3-yl)ethyl; 4-bromo-2- methylphenyi; tert-butyl; p-tolyi; and if A is -N- then R1 is selected from the group consisting of: methyl; 4-chlorophenyl; R2 is selected from the group consisting of: H, 2,5-dichlorophenyl; R3 is selected from the group consisting of: p-tolyl; 3,4- dimethoxy phenethyl.DESCRIPTION OF THE FIGURES

[0026] Figure 1. Anti-proliferative activity assay against a panel of three human cancer cell lines. Cell lines A549, HT29 and HepG2 were treated with each compound of the present invention at 50 μM for 72 h and the cell viability was assessed by MTT assay. The bars represent the percent signal relative to DMSO.

[0027] Figure 2. The dose-response curves for 72 hours in hepatocellular carcinoma (HepG2) cell lines were determined by MTT standard assay. IC50 were determined by averaging the values obtained from the fitted curves using GraphPad Prism.

[0028] Figure 3. Mice with HCC and CCR tumors established by inoculation with Hepa129 (figure 3 A) or CT-26 (Figure 3B) cells, respectively, were treated 3 times per week with 1G-55 or control prior to euthanasia. A) **p>0.01 and *P>0.05 vs. control (Mann-Whitney). B) ***p>0.001 vs. control (two-way ANOVA).

[0029] Figure 4. (A) Mouse model of liver fibrosis. Liver fibrosis was induced by intraperitoneal (i.p.) inoculation of thioacetamide (TAA) 3 times per week for 6 weeks. Treatment with 1G-55 or control was administered from week 6 until week 10, at which point the mice were euthanized. (B) Representative photomicrographs of Sirius Red-stained liver sections of mice treated with 1G-55 or DMSO, and quantification of collagen deposits as a function of Sirius Red-stained sections by morphometric analysis. ***p>0.001 vs. DMSO (Mann-Whitney).

[0030] Figure 5. (A) Dose-response curve for 72 hours in a HSC cell line (CFSC- G2) determined by a standard MTT assay. IC50 were determined by averaging the values obtained from the fitted curves using the GraphPad Prism software. (B) RNAm levels of genes associated with liver fibrosis (α-SMA and COL1A2) in the HSC cell line CFSC-G2 treated with 1G-55 (10 μM) or DMSO for 24 hours. ***p>0.001 vs. DMSO and **p>0.01 vs. DMSO (t-test).

[0031] Figure 6. (A) Scheme of acute liver failure mouse models. The mice were inoculated with acetaminophen (APAP) or concanavalin A (ConA) and treated with 1G-55 at 60 and 30 minutes, respectively. The mice were euthanized 6 hours after induction of ALF, and ALT and AST levels were measured. (B) Serum ALT and AST levels in mice with ALF treated with 1G-55. *p>0.05 vs. control (t-test).

[0032] Figure 7. (A) Scheme of acute-on-chronic liver failure mouse models. Chronic liver injury was induced by intraperitoneal administration of thioacetamide(TAA) for 10 weeks (doses of 200 mg / kg, 3 doses per week). Then, the mice were subjected to fasting for 16 hours and were injected with LPS and APAP to induce acute liver injury. The mice were treated with a single dose of control or 1G-55 one hour after induction of liver injury. (B) Liver injury was assessed by determining the serum ALT levels. ***p<0.001 vs. mice treated with control prior to acute injury and **p<0.01 vs. mice treated with control after injury.

[0033] Figure 8. Effects of 1G-55 in explants derived from patients (PDEs). 8 mm3explants of healthy human livers (healthy PDEs) or acute liver failure patient livers (ALF PDEs) were treated overnight with 1G-55 (10 μM). Transaminase levels were determined in the culture medium. (A) The absence of increased ALT release in healthy PDEs after treatment with 1G-55 suggests a lack of evident hepatotoxicity. (B) Treatment with 1G-55 enhances ALT release induced by incubation with ConA in healthy PDEs. (C) Treatment with 1G-55 reduces ALT release in ALF PDEs. The bars represent average + SEM.DETAILED DESCRIPTION OF THE INVENTION

[0034] In the context of the present invention, derivative of guanidine alpha¬ aminoacyl amide means a compound of general formula (I)A= CH, N

[0035] In the context of the present invention, “medical condition” means an abnormal anatomical or physiological condition and objective or subjective manifestations of the disease, associated with the Rho GTPase cell protein, more specifically conditions associated with increased activity or hyperactivity of the Rho GTPase cell protein family, in particular RAC1.

[0036] Rho GTPases are a family of proteins that play a key role in the transmission of signals from outside the cell to intracellular effectors, both cytoplasmatic andnuclear, that adopt different conformational states in response to the binding of GDP (guanosine diphosphate) or GTP (guanosine triphosphate). From the Rho GTPase family, RhoA, Cdc42 and RAC1 have been subjected to most research. RAC1 has been described as one of the main regulators of the actin cytoskeleton rearrangement, specifically in the formation of lamellipodia, which crucially contribute in cellular migration. An extracellular ligand, such as the epidermal growth factor (EGF) or insulin, causes the formation of lamellipodia and membrane ruffles through Rac13. Because of its important role in cytoskeleton regulation, RAC1 is considered to be involved in the regulation of processes such as endocytosis, membrane trafficking, cell morphology, adhesion, spreading and cell polarity, and in particular the regulation of cell migration. For this reason, these are considered potential therapeutic targets in medical conditions such as cancer and other conditions associated with hyperactivity of these proteins.

[0037] In the present document, the term “excipient” means an inert substance that is mixed with the active ingredient(s) to provide consistency, form, flavor or other qualities to manufactured medicaments in order to aid in their dosing and use.

[0038] In the present document, the term “adjuvant” relates to substances with different chemical structures that are used to enhance the response in patients when coadministered with the compound of the present invention.

[0039] In the present document, the term "treatment" refers to the group of actions and procedures intended to alleviate, cure or manage a disease, condition or injury. The interpretation of “treatment” may vary widely depending on the nature of the condition and may include a combination of medical, surgical, pharmacological and therapeutic methods.

[0040] The compounds of Formula (I) may contain one or more asymmetric centers and can thus occur as racemates and racemic mixtures, single enantiomers, diastereoisomeric mixtures and individual diastereoisomers. Centers of asymmetry that are present in the compounds of Formula (I) can all independently of one another have S configuration or R configuration. The compounds of Formula (I) include all possible enantiomers and diastereomers and mixtures of two or more stereoisomers, for example, mixtures of enantiomers and / or diastereomers, in all ratios. Thus, enantiomers are a subject of the disclosure in enantiomerically pure form, both as levorotatory and as dextrorotatory antipodes, in the form of racemates and in the form of mixtures of the two enantiomers in all ratios. In the case of acis / trans isomerism, the disclosure includes both the cis form and the trans form (and both the E form and the Z form) as well as mixtures of these forms in all ratios. The present disclosure is meant to comprehend all such stereoisomeric forms of the compounds of Formula (I). Where a structural formula or chemical name specifies a particular configuration at a stereocenter, the enantiomer or stereoisomer of the compound resulting from that specified stereocenter is intended. Where a structural formula of the compounds of Formula (I) indicates a straight line at a chiral center, the structural formula includes both the S and R stereoisomers associated with the chiral center and mixtures thereof.

[0041] The compounds of Formula (I) may be separated into their individual diastereoisomers by, for example, fractional crystallization from a suitable solvent, for example, methanol or ethyl acetate or a mixture thereof, or via chiral chromatography using an optically active stationary phase. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration. Vibrational circular dichroism (VCD) may also be used to determine the absolute stereochemistry. Alternatively, any stereoisomer or isomers of the compounds of Formula (I) may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known absolute configuration.

[0042] If desired, racemic mixtures of the compounds may be separated so that the individual enantiomers are isolated. The separation can be carried out by methods well known in the art, such as the coupling of a racemic mixture of compounds to an enantiomerically pure compound to form a diastereoisomeric mixture, followed by separation of the individual diastereoisomers by standard methods, such as fractional crystallization or chromatography. The coupling reaction is often the formation of salts using an enantiomerically pure acid or base. The diasteromeric derivatives may then be converted to the pure enantiomers by cleavage of the added chiral residue. The racemic mixture of the compounds can also be separated directly by chromatographic methods utilizing chiral stationary phases, which methods are well known in the art.

[0043] The inventors have designed a novel family of guanidine alpha-aminoacyl amides for the inhibition of RAC1. This includes two sub-families that differ in the nature of the group “A” (Formula I): the first one with two carbocyclic ringssubstituting the guanidine (A-CH, 1F-181, 1F-182, 1F-183, 1F-184, 1F-188, 1F-189, 1 F-190, 1G-54, 1G-55, 1G-56, 1G-57, 1G-58, 1G-59) and the second one with a pyridine ring in the terminal position of the guanidine group (A=N, 1G-130 and 1G-132).

[0044] The main object of the present invention is to provide novel compounds that are Rho GTPase inhibitor molecules, in particular RAC1 inhibitors, where said compounds have a general chemical structure of Formula IA= CH, N

[0045] where R1 is a substituent selected from the group consisting of: H; (C1-C4) alkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides may be selected from the group consisting of Cl, F, Br and I; mono- or dihydroxy aryl; 6-membered heteroaryl with one heteroatom selected from the group consisting of N, O, and S; methyl benzoate, methyl acetate, S-methyl imidazole propanoate;

[0046] where R2 is selected from the group consisting of: H (hydrogen); substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides may be selected from the group consisting of Cl, F, Br and I; mono- or dimethoxy phenethyl; benzo-D-1,3 dioxolyl, N-indolyl methyl; and

[0047] where R3 is selected from the group consisting of: H; (C1-C4) alkyl; (C5-C6) cycloalkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides may be selected from the group consisting of Cl, F, Br and I; tert¬ butyl (disubstituted propyl), mono- or dimethoxy phenethyl, N-indolyl ethyl.

[0048] In an embodiment, if in formula I A is -CH-, then R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4-chlorophenyl; 4-fluorophenyl; 3,5- dimethylphenyl; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyl; 2,5-dichlorophenyl; 1-methyl-1H-indol-3-yl; benzo[dj[1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1 H-indol-3-yl)ethyl; 4-bromo-2- methylphenyl; tert-butyl; p-tolyl.

[0049] In a further embodiment, if in formula I A is N, then R1 is selected from the group consisting of: methyl; 4-chlorophenyl; R2 is selected from the group consisting of: H, 2,5-dichlorophenyl; R3 is selected from the group consisting of: p-tolyl; 3,4-dimethoxyphenethyl.

[0050] In a preferred embodiment, if the compound derived from guanidine alphaaminoacyl amide of the present invention is represented by formula I where A is - CH-, then the combinations of R1, R2 and R3 shown in table 1 result in the following compounds: 1F-181, 1F-182, 1F-183, 1F-184, 1F-188, 1F-189, 1F-190, 1G-54, 1G-55, 1G-56, 1G-57, 1G-58, 1G-59:1F-1881F-1891G-56

[0051] In a further preferred embodiment of the present invention, if the compound derived from guanidine alpha-aminoacyl amide of the present invention is represented by formula I where A is -N, then the preferred combinations of R1, R2 and R3 are those shown in the following table 2, resulting in compounds 1G-130 and 1G-132.1G-130

[0052] In a more preferred embodiment, the compounds derived from guanidine alpha-aminoacyl amide of the present invention are listed in the following table 3:TABLE 3Compound IUPAC namemethyl 2~(N-(2~(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- 1F-181dimethylphenyl)guamdmo)benzamido)acetate N~(2-((4-bromo-2-methylphenyi)ammo)-2-oxo-1 -phenylethyl)- 3-(3-(3,5-dimethylphenyl)guanidmo)-N-(4- 1F-182hydroxyphenyi)benzamideN-(2-(t-butylamino)-2-oxo-1 -phenylethyl)~3-(3-(3,5- 1F-183dimethylphenyl)guanidino)-N-(pyndm-3-yl)benzamide2-(N-(2-(cyclohexylammQ)"2-oxQethyl)-3-(3-(3,5"F-184dimethylphenyl)guanidmo)benzamido)acetic acidN-(1 -(benzo[d][1,3]dloxol-5-yl)-2-(cyclohexylammo)-2- F-188 oxoethyl)-3-(3-(3,5-dlmethylphenyl)guanldmo)benzamidemethyl 2-(3-(3-(3,5-dimethylphenyl)guanidmo)-N-(2~((4- F-189methoxyphenethyl)ammo)-2~oxoethyl)benzamido)benzoate methyl 2-(N-(1”(^®nzoM[1,3]dioxol-5-yl)-2-(cyclohexylammo)- F-190 2-oxoethyl)-3-(3-(3!5-d!methylphenyl)giianidlno)benzamldo)-3- (1 H-lmidazel-5-yl)propanoateN-(2-((2-(1H-mdol-3-yl)ethyl)ammo)-1-(1-methyl-1H-mdol-3-yl)- G-54 2-oxoethyl)-N-(3,5-dimethylphenyl)-3-(3-(3,5- dimethylphenyl)guamdmo)benzamlde N41~(2,5“dichloropher3yn-2-oxo-2~(p~toly^mmo)ethyl)-3-(3- G-55 (3,5-dlmethylphenyOguanidmo)-N-methylbenzamldeN-(1-(2-chlorophenyl)-2-oxo~2-(P"tolylammo)ethyl)-3-(3~(3!5~ G-56dimethylphenyl)guanidmo)-N-methylbenzamide N-(4-chlorophenyl)-N~(2-((3,4-dimethoxyphenethyl)amino)~2- G-57oxQethyl)-3-(3-(3,5"dlmethylphenyl)guanldino)benzamide N-(2-((3,4-dlmethoxyphenethyl)ammo)-2~oxoethyl)-3-(3-(3,5- G-58dimethylphenyl)guanidmo)-N-(4-fluorophenynbenzamlde N~(2-((3,4-dlmethoxyphenethyl)ammo)-2-oxoethyl)-3-(3~(3.5- G-59dimethylphenyl)guanidmo)-N-(4-hydroxyphenyl)benzamide N~(4-chlorophenyO~N-(2-((3.4-dimethoxyphenethyl)amino)-2- G-130oxQethyl)-3-(3"(4,6"dlmethylpyndm-2-yl)guanidino)benzamide G-132 N~(1-(2,5-dichlorophenyl)-2-oxo~2-(p-tolylammo)ethyl)-3-(3-(4, 6-dimethylpyridin-2-yl)guanidino)-N-methyl benzamide

[0053] A further object of the present invention is to provide a compound selected from the group consisting of:1G-55 1F-1821G-581F-190 1F-1841F-1881G-132 and or a pharmaceutically acceptable salt thereof.

[0054] A further object of the present invention is to provide a compound1G’55or a pharmaceutically acceptable salt thereof.

[0055] A further object of the present invention is to provide a pharmaceutical composition comprising at least one compound derived from guanidine alpha¬ aminoacyl amide of the present invention and at least one compound selected from the group consisting of an excipient, an adjuvant, or a combination thereof.

[0056] In addition, said pharmaceutical composition of the present invention may be used for the treatment of medical conditions associated with increased activity or hyperactivity of the RhoGTPase protein, in particular RAC1. In particular embodiment of the invention, said medical condition to be treated is associated with cell proliferation, such as liver cancer, hepatocellular carcinoma, gastrointestinal cancer, colorectal cancer, pancreatic cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma, and the like. In a further preferred embodiment of the present invention, the pharmaceutical composition of the present invention may be used for the treatment of medical conditions such as acute liver failure, acute-on-chronic liver failure (ACLF), and liver fibrosis.

[0057] A further object of the present invention is to provide a method for the treatment, in a subject in need thereof, of a medical condition associated with hyperactivity of the RAC1 protein, or a change in the physiological activity thereof, by administering a compound derived from guanidine alpha-aminoacyl amide of the present invention in a pharmacologically acceptable amount.

[0058] The compounds derived from guanidine alpha-aminoacyl amide of the present invention exhibit in vitro half-maximal inhibitory concentration values (IC50), in assays with hepatocellular carcinoma (HepG2), colorectal carcinoma (HT29), lung cancer (A549) and breast cancer (A375) cell lines, that are acceptable to be used in the treatment of medical conditions associated with RAC1, as shown in ExampleSaid example demonstrates that the compounds of the present invention have the ability to inhibit the interaction between RAC1 and its GEF (TIAM1) and proliferation in colorectal carcinoma (HT29) and lung cancer (A549) by an alpha assay and a standard MTT assay, respectively.

[0059] Additionally, the results obtained in animal models of liver injury demonstrate that the compounds derived from guanidine alpha-aminoacyl amide of the present invention have a surprising anti-fibrotic effect. For example, the treatment with 1G-55 markedly reduced collagen deposits in the livers of mice with fibrosis (Figure 4). The compounds derived from guanidine alpha-aminoacyl amide of the present invention also reduced the expression of the pro-fibrogenic markers CollA2 and a-SMA (Figure 5B) in hepatic stellate cells (HSCs) and hepatic macrophages.

[0060] The effects of the compounds derived from guanidine alpha-aminoacyl amide of the present invention in cell viability and the phenotype of hepatic stellate cells (HSCs), which play key role in the production of type I collagen during the fibrogenic process, were also evaluated. As shown in Figure 5A, for example, 1G- 55 markedly reduces the survival of HSCs in vitro with an IC50 value similar to the value observed in cancer cells. In addition, 1G-55 reduces the expression of the pro- fibrogenic markers CollA2 and a-SMA (Figure 5B) in HSCs.

[0061] The inventors measured alanine aminotransferase (ALT) levels as an indication of liver injury, finding a significant reduction in ALT when applying the treatment with the compounds derived from guanidine alpha-aminoacyl amide of the present invention, as shown with the compound 1G-55. This may reflect a reduction in liver damage in acute-on-chronic liver failure. Likewise, it was shown that the treatment with 1 G-55 significantly reduced the degree of liver damage, evaluated by the measurement of AST and ALT in both ALF models, as shown in figure 6.

[0062] A further object of the present invention is to provide a method for treating a liver disease to a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound selected from the group consisting of:1G-57 and, or a pharmaceutically acceptable salt thereof.

[0063] A further object of the present invention is to provide a method for treating a liver disease to a subject in need thereof comprising administering to the subject atherapeutically effective amount of a compoundor a pharmaceutically acceptable salt thereof.

[0064] A further object of the present invention is to provide a method for preparing the guanidine alpha-aminoacyl amide compounds of Formula I where A=CH, where said method comprises the steps of 1)- obtaining the chemical precursor N-(f- butoxycarbonyl)-N'-(3,5-dimethylphenyl)thiourea (1D-195); 2)- from said compound, obtaining the intermediate 3-(2-(t-butoxycarbonyl)-3-(3,5- dimethylphenyl)guanidino)benzoic acid (1F-123); 3)- obtaining the guanidine alpha- aminoacyl amide compound of the present invention, from an Ugi multicomponent reaction.

[0065] A further object of the present invention is to provide a method for preparing the guanidine alpha-aminoacyl amide compounds of Formula I of the present invention, where A=N, where said method comprises the steps of 1)- obtaining the chemical precursors 3-amino-N-(1 -(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-N-methylbenzamide (1G-125) and 3-amino-N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)benzamide (1G-126) from an Ugi reaction followed by catalytic hydrogenation; 2)- from said compound, obtaining the guanidine alpha-aminoacyl amide compounds of the present invention, through a coupling reaction with cyanamide.

[0066] A further object of the present invention is to provide a method for the synthesis of a guanidine alpha-aminoacyl amide compound of the present invention where if A is N said method comprises the following steps: i) obtaining the chemical intermediate 3-amino-N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-N- methylbenzamide or 3-amino-N-(4-chlorophenyl)-N-(2-((3,4- dimethoxyphenethyl)amino)-2-oxoethyl)benzamidefrom a solution of 3-nitrobenzoic acid, an aldehyde, an amine in MeOH, adding an isonitryl by Ugi reaction followed by catalytic hydrogenation; ii) obtaining the chemical intermediate N-(4,6- dimethylpyridin-2-yl)cyanamide from a solution of N-(4,6-dimethyl-2-pyridyl)thiourea, a KOH solution, and adding a 1,5 M lead acetate solution (Process for the preparation of the compound N-(3,5-dimethylphenyl)-N’(2-trifluoromethylphenyl)guanidine., US20180244608 A1); iii) coupling each intermediate obtained in step i) with the intermediate from step ii), under suitable conditions, to provide N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p- tolylamino)ethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)-N-methylbenzamide and N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(4,6- dimethylpyridin-2- yl)guanidino)benzamide, respectively.

[0067] The present invention is described in greater detail in the following examples, which should not be considered as a limitation to the scope of the invention. The invention is not limited by the illustrative examples presented hereinafter.EXAMPLES

[0068] All chemical compounds were acquired from commercial sources or were synthesized. The solvents used were of analytical grade or higher.

[0069] Use of anhydrous solvents and moisture-sensitive reagents: los solvents used were of analytical grade or higher. THF was distilled with sodium-benzophenone prior to use, and CH2CI2was refluxed with P2O5, prior to distillation and storage on molecular sieves (4 A). All reactions involving moisture-sensitive solvents or reagents were carried out in dry glassware and under nitrogen or argon atmosphere.

[0070] Thin layer chromatography: silica gel F2540,2 mm thick aluminum sheets (Merck) were used. The solvent system used (ascending method) are indicated in each case. The development was carried out by: (a) exposure to ultraviolet light (A = 254 nm); (b) immersion in Mo7O24(NH4)60.04 M, Ce(SO4)23 mM in solution H2SO4-H2O (9:1) followed by heating; (c) immersion in ninhydrin 0.1% w / v solution in ethanol followed by heating; (d) exposure to molecular iodine.

[0071] Column chromatography: it was carried out with silica gel 60230-400 mesh (Merck). Eluting solvents are indicated in each case.

[0072] Flash chromatography: it was carried out in a CombiFlash Teledyne Isco instrument with disposable RediSep silica gel columns. Solvents and elution conditions are indicated in each case.

[0073] High resolution liquid chromatography: it was carried out in a Waters Alliance 2695 instrument equipped with a photodiode array (PDA) detector.

[0074] Melting points: these were determined with an Electrothermal IA9000series apparatus and are not corrected.

[0075] Nuclear magnetic resonance: nuclear magnetic resonance spectra were recorded with Bruker Fourier 300 (operating at 300 MHz for1H and 75 MHz for13C), Avance DPX 400 (operating at 400 MHz for1H and 100 MHz for13C) spectrometers at 303 K. The following deuterated solvents were used, as indicated in each case: CDCh, CD3OD, DMSO-d6. Chemical shifts are indicated in parts per million (ppm) relative to tetramethylsilane (TMS) signal. Coupling constants are expressed in Hz. Signals are described as s (singlet), d (doublet), t (triplet), q (quartet), quint, (quintet), dd (double doublet), ddd (double double doublet), dt (double triplet), tt (triple triplet), dist. t (distorted triplet), bs (broad singlet) and m (multiplet).

[0076] In some cases, the assignment of1H or13C magnetic resonance spectral signals was carried out using a combination of one- and two-dimensional techniques, such as heteronuclear1H-13C (HSQC and HMBC) and / or homonuclear (COSY-45 and ROESY1H-1H) correlation experiments.

[0077] Mass spectrometry: electronic impact (El) and eletrospray (ESI) ionization techniques were used. Low resolution El spectra were acquired in a QP2010 ultra spectrometer at 70 eV. ESI spectra were acquired in a Quattro Premier XE spectrometer. High resolution mass spectra were acquired in CIBION (CONICET) with a Xevo G2S Q-TOF spectrometer calibrated in the m / z 50-1200 interval with a sodium formate 0,5 mM solution prepared in 90:10 2-propanol:agua v / v. The data were corrected during acquisition using a reference compound (LockSpray).Scheme of the synthetic route for guanidine alpha-aminoacyl amide compounds where A=CH of the present invention.1F-181, 1F-182, 1F183, 1F-184, 1F-188, 1F-189,1F-123Example 1: Synthesis of intermediate N-(-butoxycarbonyl)-N'-(3,5-dimethylphenyl)thiourea (1D-195)

[0078] To a solution of thiourea (521 mg; 6.84 mmol) in 117 mL of anhydrous THF 60% NaH (1.23 g; 30.75 mmol) was added portion-wise at 0 °C under argon atmosphere. It was warmed to room temperature and reacted for 45 min. Then it was placed in an ice bath at 0 °C and di-tert-butyl dicarbonate (3.3 mL, 14.36 mmol) was added dropwise. The reaction mixture was warmed to room temperature and reacted overnight. Then it was cooled to 0 °C and 60% NaH (349.2 mg; 8.73 mmol) was added portion-wise, and reacted for 1 h before adding trifiuoroacetic anhydride (1.0 mL; 7.08 mmol) dropwise. The mixture was reacted at 0 °C for another 1 h to form the thioacyiating agent and 3,5-dimethylaniline (1.23 mL; 9.87 mmol) was added dropwise. Then it was warmed to room temperature and reacted overnight. The reaction was quenched by the addition of H2O (120 mL) at 0 °C and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with NaCI saturated solution (30 mL), dried (Na2SO4) and the solvent was evaporated. The residue was purified by flash chromatography (silica gel) eluted with hexane-EtOAc gradient followed by recrystallization on MeOH-H2O to provide 774 mg (40%) of ID- 195 as a white solid (35).

[0079] Rf 0.26 (95:5 hexane: EtOAc). Mp 122 °C.1H NMR (300 MHz, CDCI3): 5 11.39 (s, 1H), 8.01 (s, 1H), 7.24 (s, 2H), 6.89 (s, 1H), 2.33 (s, 6H), 1.53 (s, 9H).13CNMR (75 MHz, CDCI3): 5 178.2, 152.0, 138.7, 137.6, 128.6, 122.1, 84.6, 28.1, 21.4. EM-ESI m / z 281 ([M+H]+).Example 2: Synthesis of intermediate 3-(2-(-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid (1F-123)

[0080] To a solution of 1D-195 (1.56 g; 5.56 mmol), benzyl 3-aminobenzoate (1.70 g; 7.48 mmol) and triethylamine (0.9 mL; 6.46 mmol) in 60 mL of anhydrous CH2CI2EDCI (2.48 g; 12.94 mmol) was added at 0 °C and under argon atmosphere. The residue was purified by flash chromatography (silica gel) eluted with hexane-EtOAc gradient. The reaction product (2.24 g; 4.73 mmol) was dissolved in 50 mL of EtOAc and treated with hydrogen at 2 bar in the presence of 10% Pd on carbon (227 mg) for 24 h at room temperature. The mixture was filtered through a Celite bed and the solvent was evaporated to provide 1.82 g of 1F-123 (100%) as a white solid.

[0081] Rf 0.50 (1:1 hexane: EtOAc). Mp 118-119 °C.1H NMR (300 MHz, CDCI3): 6 10.75 (bs, 3H.), 7.77 (s, 1H), 7.72 (d, = 7.5 Hz, 1H), 7.28 (bs, 1H), 7.20 (t, J= 7.8 Hz, 1H), 6.71 (s, 2H), 6.58 (s, 1H), 2.11 (s, 6H), 1.54 (s, 9H).13C NMR (75 MHz, CDCI3): 5 171.8, 156.3, 149.3, 138.8, 138.5, 137.2, 133.4, 128.6, 126.9, 126.2, 126.1, 124.2, 121.0, 83.0, 28.2, 21.2. EM-ESI 384 ([M+H]+).General procedure A for the synthesis of guanidine alpha-aminoacyl amide compounds where A=CH by Ugi reaction

[0082] To a solution of 1F-123 (1,0 eq.) in MeOH, an amine (1,0 eq.) and an aldehyde (1,0 eq.), isonitryl (1,0 eq.) was added while stirring, and reacted for 2 to 7 days at room temperature. Then the solvent was evaporated and the residue was purified by chromatography (silica gel) to provide a product that was treated with 15.5% HCI in isopropanol for 4-5 h at room temperature. Then the pH was adjusted to 8-9 by the addition of NaHCO3saturated solution at 0 °C. The mixture was extracted with CH2CI2(x 5), the combined organic layers were dried (Na2SO4) and the solvent was evaporated. The residue was purified by column chromatography (silica gel) to provide the desired products 1F-181, 1F-182, 1F-183, 1F-184, 1F-188, 1F-189, 1F-190, 1G-54, 1G-55, 1G-56, 1G-57, 1G-58, 1G-59.

[0083] Exemplary syntheses of the compounds derived from guanidine alpha¬ aminoacyl amide where A is CH- are described below:Example 3: Synthesis of N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(3,5-dimethylphenyl)guanidino)-N-methylbenzamide (1G-55)1G-55

[0084] Following general procedure A, 2,5-dichlorobenzaldehyde (88 mg; 0.50 mmol), methylamine hydrochloride (35 mg; 0.52 mmol), 1F-123 (196 mg; 0.51 mmol), triethylamine (70 pL; 0.50 mmol) and 4-methylphenyl isonitrile (95 mg; 0.81 mmol) were reacted in 1 mL of MeOH for 6 days. The reaction was purified by column chromatography (silica gel) eluted with hexane-EtOAc to provide 67 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide 42 mg (14 %, total) of 1G-55 as a white solid.

[0085] Rf 0.25 (95:5:0.5 CH2CI2-MeOH-NH3). Mp 156-157 °C.1H NMR (300 MHz, MeOD): 57.56 (d, J = 8.4 Hz, 1H), 7.48 (d, J = 8.1 Hz, 2H), 7.43 (s, 1H), 7.40 (d, J = 7.9 Hz, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.30 (s, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.15 (d, J = 8.3 Hz, 2H), 7.08 (d, J = 7.4 Hz, 1H), 6.80 (maj) and 6.76 (mln) (rot., s, 2H), 6.65 (s, 1H), 6.48 (maj) and 5.76 (min) (rot., s, 1H), 2.87 (min) and 2.80 (maj) (rot., s, 3H), 2.31 (maj) and 2.28 (min) (rot., s, 3H), 2.24 (s, 6H).13C NMR (75 MHz, MeOD): 5 175.4 (min) and 174.6 (maj) (rot.), 169.4 (min) and 169.2 (maj) (rot.), 152.9 (maj) and 152.6 (min) (rot.), 147.8 (min) and 147.5 (maj) (rot.), 144.0 (min) and 143.9 (maj) (rot.), 139.7 (maj) and 139.6 (min) (rot.), 137.7 (min) and 137.3 (maj) (rot.), 136.8, 136.2 (maj) and 135.9 (min) (rot.), 135.7 (min) and 135.4 (maj) (rot.), 135.2 (maj)and 135.1 (min) (rot.), 134.3, 132.8 (min) and 132.7 (maj) (rot.), 131.7 (min) and 131.5 (maj) (rot.), 131.4, 130.5, 130.4, 125.6, 125.4 (min) and 125.3 (maj) (rot.), 121.8 (maj) and 121.7 (min) (rot.), 121.6 (maj) and 121.5 (min) (rot.), 121.3, 120.8 (maj) and 118.3 (min) (rot.), 65.7 (min) and 61.4 (maj) (rot.), 36.4 (maj) and 31.9 (min) (rot.), 21.5, 21.0. EM-ESI m / z 588 ([M+H]+).Example 4: Synthesis of N-(2-((4-bromo-2-methylphenyl)amino)-2~oxo-1~ phenylethyl)-3-(3-(3,5-dimethylphenyl)gua dmo)-N-(4- hydroxyphenyljbenzamlde (1 F-182)1 F-182

[0086] Following general procedure A, to a solution of benzaldehyde (13 pL; 0.13 mmol), 4-aminofenol (15 mg; 0.14 mmol) and 1 F-123 (51 mg; 0.13 mmol) in 0.5 mL of MeOH, 4-bromo-2-methylphenyl isonitryl (25 mg; 0.13 mmol) was added and the mixture was reacted for 3 days at room temperature. Then the solvent was evaporated and the reaction was purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH to provide 21 mg of a product that was treated with HCI in iPA (0.5 mL) for 4 h at room temperature, and then the pH was adjusted to 8 by the addition of NaHCO3saturated solution at 0 °C. The mixture was extracted with CH2CI2(5 x 2.5 mL), the combined organic layers were dried (Na2SO4and the solvent was evaporated. The residue was purified by flash chromatography (silica gel) eluted with CH2CI2-MeOH gradient to provide 13 mg (15%, total) of 1 F-182 as a white solid.

[0087] Rf 0.32 (6:4 hexane-EtOAc). Mp 152-154 °C.1H NMR (300 MHz, CDCI3 / MeOD): 0 7.69 (d, J = 8.1 Hz, 1H), 7.40-7.24 (m, 9H), 7.20 (s, 1H), 7.09 (d, J = 6.6 Hz), 6.98 (s, 1H), 6.90 (s, 2H), 6.82 (dist. d, 2H), 6.50 (d, J = 8.1 Hz, 2H),6.25 (s, 1H), 2.33 (s, 6H), 2.03 (s, 3H).13C NMR (75 MHz, CDCI3 / MeOD): 5 171.1, 168.1, 156.8, 154.4, 140.3, 137.9, 135.8, 134.4, 133.6, 133.2, 132.4, 131.6, 131.5, 130.4, 130.1, 129.9, 129.6, 129.3, 129.0, 127.4, 126.7, 125.0, 124.7, 123.4, 118.6, 115.6, 67.4, 21.2, 17.3. EM-ESI m / z 676 ([M+H]+).Example 5: synthesis of N-(2-(t-butylam o)~2-oxo-1~phenylethyl)-3-(3-(3,5- dimethylphenyl)guamdmo)-N-(pyridin-3-yl)benzamicle (1 F- 83)1F-183

[0088] Following general procedure A, benzaldehyde (13 pL; 0.13 mmol), 3-aminopyridine (13 mg; 0.14 mmol), 1 F-123 (51 mg; 0.13 mmol) and t-butyl isonitryl (16 µL; 0.14 mmol) in 0.5 mL of MeOH were reacted for 5 days. The reaction was purified by column chromatography (silica gel) eluted with hexane-EtOAc to provide 35 mg of a product that was treated with HCI in IPA (0.75 mL) and purified by flash chromatography (silica gel) eluted with CH2CI2-MeOH gradient to provide 19 mg (27 %, total) of 1F-183 as a white solid.

[0089] Rf 0.41 (9:1 CH2CI2-MeOH). Mp 138 °C.1H NMR (300 MHz, CDCI3): 58.14 (bs, 1H), 8.07 (s, 1H), 7.50 (bs, 1H), 7.28-7.14 (m, 9H), 6.97 (s, 2H), 6.87 (s, 2H), 6.21 (s, 1H), 5.72 (s, 1H), 2.32 (s, 6H), 1.34 (s, 9H).13C NMR (75 MHz, CDCI3): 6 170.0, 168.4, 154.6 151.4, 141.8, 140.5, 138.4, 137.5, 136.9, 134.7, 133.9, 133.8, 130.4, 130.15, 130.08, 129.2, 129.0, 127.5, 126.9, 125.0, 123.4, 65.9, 52.1, 28.7, 21.3. EM-ESi m / z 549 ([M+HJ+).

[0090] Example 6: synthesis of methyl 2-(3-(3-(3,5-dimethylphenyl)guanidmo)~ N-(2-((4-methoxyphenethyl)am o)-2-oxoethyl)benzamido)benzoate (1 F-189).0.1F-189

[0091] Following general procedure A, 37% formaldehyde (38 pL; 0.51 mmol), methyl antranilate (65 pL; 0.50 mmol), 1F-123 (191 mg; 0.50 mmol) and 4- methoxyphenethyl isonitryl (81 mg; 0.50 mmol) in 0.5 mL of MeOH were reacted for 7 days. The reaction was purified by column chromatography (silica gel) eluted with hexane-EtOAc to provide 89 mg of a product that was treated with HCI in IPA (1.2 mL) and purified by column chromatography (silica gel) eluted with CH2CI2-MeOH to provide 45 mg (15 %, total) of 1 F-189 as a white solid.

[0092] R / 0.47 (9:1 CH2CI2-MeOH). Mp:118-120 °C.1H NMR (300 MHz, CDCI3): 0 7.62 (d, = 7.8 Hz, 1H), 7.55 (m, 2H), 7.43 (d, = 7.9 Hz, 1H), 7.27 (t, J = 7.6 Hz, 1H), 7.19 (d, = 8.5 Hz, 1H), 7.17 (s, 1H), 7.12 (t, J = 7.7 Hz, 1H), 7.03 (d, = 8.5 Hz, 2H), 6.95 (s, 1H), 6.91 (d, = 7.8 Hz, 1H), 6.85 (s, 2H), 6.73 (d, = 8.6 Hz, 2H), 4.53 (d, 16.3 Hz, 1H), 4.29 (d, J = 16.1 Hz, 1H), 3.73 (s, 3H), 3.67 (s, 3H), 3.55- 3.35 (m, 2H), 2.70 (t, J = 6.9 Hz, 2H), 2.30 (s, 6H).13C NMR (75 MHz, CDCh): 5 169.6, 168.1, 166.8, 158.1, 154.6, 142.8, 140.4, 136.3, 134.8, 134.2, 134.0, 131.7 131.0, 129.9, 129.8, 129.5, 128.3, 127.9, 127.5, 127.42, 127.40, 125.5, 123.2, 114.0, 55.3, 54.9, 52.9, 40.9, 34.5, 21.3. EM-ESI m / z 608 ([M+H]+).Example 7: synthesis of N-(1-(2-chlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3- (3-(3,5-dimethylphenyl)guanidino)- -methylbenzamide (1G-56)1G-56

[0093] Following general procedure A, 2-chlorobenzaldehyde (56 pL; 0.50 mmol), methylamine hydrochloride (36 mg; 0.53 mmol), 1F-123 (195 mg; 0.51 mmol), triethylamine (70 L, 0.50 mmol) and 4-methylphenyl isonitrile (65 mg; 0.55 mmol) in 1 mL of MeOH were reacted for 3 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-EtOActo provide 96 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide 51 mg (18 %, total) of 1G-56 as a white solid.

[0094] Rr0.30 (95:5:0.5 CH2CI2-MeOH-NH3). Mp 149 °C.1H NMR (300 MHz, MeOD): 07.57 (d, = 7.4 Hz, 1H), 7.50 (d, = 8.2 Hz, 2H), 7.45-7.35 (m, 4H), 7.32 (maj) and 7.29 (min) (rot., s, 1H), 7.27 (d, = 7.9 Hz, 1H), 7.14 (d, = 8.2 Hz, 2H), 7.09 (d, J = 8.3 Hz, 1H), 6.84 (maj) and 6.78 (min) (rot., s, 2H), 6.71 (s, 1H), 6.54 (maj) and 5.79 (min) (rot., s, 1H), 2.84 (min) and 2.76 (maj) (rot., s, 3H), 2.31 (maj) and 2.26 (min) (s, 3H), 2.26 (s, 6H).13C NMR (75 MHz, MeOD): 6 175.2 (min) and 174.3 (maj) (rot.), 170.1 (min) and 169.9 (maj) (rot.), 153.5 (maj) and 153.3 (min) (rot.), 146.1 (min) and 145.6 (maj) (rot.), 143.2 (min) and 142.4 (maj) (rot.), 140.0 (maj) and 139.9 (min) (rot.), 138.1 (min) and 137.8 (maj) (rot.), 137.4 (min) and 137.0 (maj) (rot.), 136.8 (maj) and 136.6 (min) (rot.), 135.5 (min), and 135.2 (maj) (rot.), 134.2 (maj) and 133.9 (min) (rot.), 131.9 (min) and 131.8 (maj) (rot.), 131.7 (min) and 131.6 (maj) (rot.), 131.3, 130.6, 130.4, 128.6 (maj) and 128.5 (min) (rot.), 126.4 (maj) and 126.2 (min) (rot.), 125.7 (min) and 125.6 (maj) (rot.), 122.6 (maj) and 122.2 (min) (rot.), 122.3 (maj) and 122.1 (min) (rot.), 121.3, 121.2, 66.1 (min) and 61.6 (maj) (rot.), 36.3 (maj) and 31.9 (min) (rot.), 21.5, 21.0. EM-ESI m / z 554 ([M+H]+).Example 8: synthesis of N-(2-((2-(1H-mdol-3~yl)ethyl)amino)-1~(1-methyi-1H- indol-3-yl)-2-oxoethyl)-N-(3,5-dimethylphenyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamide (1G-54)1G-54

[0095] Following general procedure A, 1-methylindol-3-carboxaldehyde (80 mg; 0.50 mmol), 3,5-dimethylaniline (62 pL; 0.50 mmol), 1F-123 (193 mg; 0.50 mmol) and 3-(2-isocianoethyl)-1H-indole (86 mg; 0.51 mmol) in 1 mL of MeOH were reacted for 6 days. The reaction was purified by column chromatography (silica gel) eluted with hexane-EtOAc to provide 63 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2CI2-MeOHto provide 14 mg (4%, total) of 1G-54 as a white solid.

[0096] R / 0.21 (95:5 CH2CI2-MeOH). Mp 195 °C.1H NMR (300 MHz, MeOD): 37.51 (d, J = 7.9 Hz, 1 H), 7.40 (d, J = 7.4 Hz, 1 H), 7.35 (s, 1 H), 7.31 (d, J = 8.7 Hz, 1 H), 7.28 (d, = 8.1 Hz, 1H), 7.27-7.17 (m, 3H), 7.14 (t, J = 7.7 Hz, 1H), 7.07 (t, J = 7.3 Hz, 1H), 7.00 (s, 1H), 6.94 (t, 7.8 Hz, 1H), 6.91 (s, 2H), 6.87 (s, 1H), 6.70 (s, 1H), 6.58 (s, 1H), 6.54 (bs, 2H), 6.53 (s, 1H), 3.76-3.67 (m, 1H), 3.54 (s, 3H), 3.53-3.44 (m, 1H), 2.96 (t, J = 6.9 Hz, 2H), 2.33 (s, 6H), 1.92 (s, 6H).13C NMR (75 MHz, CDCI3): 6 172.2, 171.8, 156.1, 141.5, 141.1, 139.6, 138.9, 138.1, 136.3, 135.9, 132.0, 130.5, 130.2, 129.8, 129.2, 129.0, 128.7, 128.1, 126.9, 125.6, 123.8, 123.6, 122.9, 122.3, 120.6, 119.6, 119.3, 113.0, 112.2, 110.5, 108.6, 60.0, 41.6, 32.8, 26.0, 21.3, 20.9. EM-ESI m / z 716 ([M+H]1’).Example 9: synthesis of N-(4-chlorophenyl)-N-(2~((3,4- dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamide (1G-57)1G-57

[0097] Following general procedure A, 37% formaldehyde (37 pL; 0.50 mmol), 4-chloroaniline (65 mg; 0.51 mmol), 1F-123 (193 mg; 0.50 mmol) and 3,4- dimethoxyphenethyl isonitryl (97 mg; 0.51 mmol) in 1 mL of MeOH were reacted for 2 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-EtOAc to provide 73 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2CI2-MeOH- NH3to provide 44 mg (14%, total) of 1G-57 as a white solid.

[0098] Rf 0.23 (95:5:0.5 CH2CI2-MeOH-NH3). Mp 165-166 °C.1H NMR (300 MHz, CDCI3): 67.14-7.05 (m, 3H), 7.10 (d, 8.8 Hz, 2H), 6.93 (d, 8.6 Hz, 2H), 6.85 (d, = 7.3 Hz, 1H), 6.78-6.68 (m, 7H), 5.50 (bs, 3H), 4.39 (s, 2H), 3.81 (s, 3H), 3.79 (s, 3H), 3.48 (q, J = 6.2 Hz, 2H), 2.74 (t, J = 6.9 Hz, 2H), 2.24 (s, 6H).13C NMR (75 MHz, CDCI3): 6170.8, 168.4, 150.7, 149.0, 147.7, 143.2, 142.2, 140.2, 139.4, 135.7, 132.8, 131.3, 129.5, 129.2, 128.4, 126.5, 125.1, 123.7, 123.1 121.0, 120.8, 112.0, 111.5, 55.9, 54.6, 40.7, 35.0, 21.4. EM-ESI m / z 614 ([M+H]+).Example 10: synthesis of N-(2-((3,4-dimethoxyphenethyl)ammo)-2-oxoethyl)- 3-(3-(3,5-dimethylphenyl)guanidmo)-N-(4-fluoropheny benzamide (1G-58)

[0099] Following general procedure A, 37% formaldehyde (37 pL; 0.50 mmol), 4- fluoroaniline (48 pL; 0.51 mmol), 1F-123 (171 mg; 0.45 mmol) and 3,4-dimethoxyphenethyl isonitryl (97 mg; 0.51 mmol) in 1 mL of MeOH were reacted for 2 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-EtOAc to provide 250 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2CI2-MeOH- NH3to provide 68 mg (61%, total) of 1G-58 as a white solid.

[0100] Rf 0.28 (95:5:0.5 CH2CI2-MeOH-NH3). Mp: 123-124 °C.1H NMR (300 MHz, CDCI3): 67.23 (s, 1H), 7.13 (d, J = 8.0 Hz, 1H), 7.06-6.97 (m, 4H), 6.83 (d, J = 8.2 Hz, 2H), 6.80 (d, J = 8.6 Hz, 1H), 6.74-6.66 (m, 6H), 6.40 (bs, 3H, NH), 4.38 (s, 2H), 3.79 (s, 3H), 3.77 (s, 3H), 3.43 (m, 2H), 2.71 (t, J = 6.9 Hz, 2H), 2.22 (s, 6H).13C NMR (75 MHz, CDCI3): 6 170.6, 168.4, 161.1 (d, JC-F = 246.6 Hz), 151.5, 148.9, 147.6, 140.8, 139.5, 139.4, 138.9, 136.0, 131.3, 129.00, 128.94 (d, JC-F = 8.6 Hz), 127.0, 125.0, 124.3, 123.4, 121.3, 120.7, 116.2 (d, JC-F = 22.5 Hz), 112.0, 111.4, 55.9, 54.6, 40.8, 35.0, 21.3. EM-ESI m / z 598 ([M+H]+)Example 11: synthesis of N-(2-((3.4-dimethoxyphenethyi)am o)-2-oxoethylL 3-(3-(3,5-dimethylphenyi)guanidirro)-N-(4-hydroxyphenyl)ber8zamide (1G-59)1G-59

[0101] Following general procedure A, 37% formaldehyde (37 pL; 0.50 mmol), 4- aminofenol (55 mg; 0.51 mmol), 1F-123 (185 mg; 0.48 mmol) and 3,4- dimethoxyphenethyl isonitryl (100 mg; 0.52 mmol) in 1 mL of MeOH were reacted for 2 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-EtOAc to provide 207 mg of a product that was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2CI2- MeOH-NH3to provide 64 mg (47%, total) of 1G-59 as a white solid.

[0102] Rf 0.20 (95:5:0.5 CH2CI2-MeOH-NH3). Mp 190 °C.1H NMR (300 MHz, CDCI3): 67.04-6.95 (m), 6.75-6.66 (m, 8H), 6.50 (d, J = 8.0 Hz), 6.14 (bs, 4H), 4.36(s, 2H), 3.76 (s, 3H), 3.75 (s, 3H), 3.44 (m, 2H), 2.71 (dist t, 2H), 2.19 (s, 6H).13C NMR (75 MHz, CDCI3): 6 171.0, 169.2, 156.8, 152.0, 149.0, 147.7, 141.3, 139.5, 138.3, 136.6, 134.6, 131.3, 129.4, 128.3, 127.2, 125.5, 124.8, 123.8, 121.5Example 12: synthesis of methyl 2-( -(2-(cyclohexylam o)-2-oxoethyl)-3~(3- (3,5-dimethylphenyl)guamdmo)benzamido)acetate (1F-181)1F-181

[0103] Following general procedure A, 37% formaldehyde (21 pL; 0.28 mmol), methyl glycinate hydrochloride (35 mg; 0.28 mmol), 1F-123 (104 mg; 0.27 mmol), 2.5 N KOH in MeOH (110 pL; 0.28 mmol) and cyclohexyl isonitrile (34 pL; 0.26 mmol) in 1 mL of MeOH were reacted for 2 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-MeOH to provide 80 mg of the product methyl (2-(3-(2-(t-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)-N-(2- (cyclohexylamino)-2-oxoethyl)benzamido)acetate (1 F-139) (100% purity by HPLC and [M+H]+ (ESI) = 594.), which was treated with HCI in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2CI2-MeOH to provide 31 mg (38%, total) of 1F-181 as a white solid.

[0104] Rf 0.60 (9:1 CH2CI2: MeOH). Mp 120-121 °C.1H NMR (300 MHz, CDCI3): 6 7.91 (d, J = 6.4 Hz, 1 H), 7.47-7.21 (m, 4H), 6.95 (s, 1 H), 6.88 (s, 2H), 4.24 (maj) and 3.98 (min) (rot., s, 2H), 4.09 (min) and 3.98 (maj) (rot., s, 1H), 3.81 (maj) and 3.71 (min) (rot., s, 3H), 3.43 (bs, 1H), 2.31 (s, 6H), 1.86-1.68 (m, 4H), 1.60-1.57 (m, 1H), 1.33-1.07 (m, 5H).13C NMR (75 MHz, CDCI3): 0 171.4, 170.7 (maj) and 170.5 (min) (rot.), 167.7 (maj) and 167.0 (min) (rot.), 155.1 (maj) and 154.7 (min) (rot.), 140.4, 135.5, 134.9, 134.0, 130.8, 129.9, 128.0, 127.1 (maj) and 125.1 (min) (rot.), 123.7 (maj) and 123.2 (min) (rot.), 123.4, 55.7 (maj) and 52.5 (min) (rot.), 53.0, 51.6 (min) and 50.7 (maj) (rot.), 48.9 (maj) and 48.5 (min) (rot.), 32.8 (min) and 32.5 (maj) (rot.), 25.4, 24.8, 21.3. EM-ESI m / z 494 ([M+H]+).Example 13: synthesis of methyl 2-(N-(1~(benzo[d][1.3]dioxol~5-yn-2- (cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidmo)benzamido)-3-(1H4midazol-5-yl)propanoate (1F- 190)1 F-190

[0105] Following general procedure A, piperonaldehyde (74 mg; 0.49 mmol), L-histidine methyl esther dihydrochloride (122 mg; 0.50 mmol), 1 F-123 (190 mg; 0.50 mmol), triethylamine (139 pL; 1.00 mmol) and cyclohexyl isonitrile (65 pL; 0.49 mmol) in 1 mL of MeOH were reacted for 6 days. The reaction was purified by column chromatography (silica gel) eluted with CH2CI2-EtOActo provide 41 mg of a product that was treated with HCi in IPA (0.5 mL) and purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide 22 mg (7%, total) of 1 F-190 as a white solid.

[0106] Rf 0.37 (CH2CI2-MeOH-NH3, 9:1:0.1). Mp 224 °C.1H NMR (300 MHz, CDCI3): 67.67 (s, 1H), 7.33 (bs, 2H), 6.97 (s, 1H), 6.89 (bs, 2H), 6.84 (s, 3H), 6.80 (d, J = 7.7 Hz), 6.74 (d, J = 7.1 Hz, 1H), 6.55 (s, 1H), 6.01 (s, 2H), 4.79 (s, 1H), 4.47 (m, 1H), 3.69 (m, 1H), 3.23 (d, J = 15.0 Hz, 1H), 3.14 (s, 3H), 2.91 (d, J = 15.2 Hz, 1H), 2.30 (s, 3H), 1.96 (m, 1H), 1.78 (m, 1H), 1.67-1.57 (m, 3H), 1.35-1.10 (m, 5H).13C NMR (75 MHz, CDCI3): δ 171., 169.5, 168.5, 152.5, 148.1, 147.9, 139.8, 136.1, 135.2, 135.1, 129.5, 127.7, 125.9, 124.1, 123.7, 122.5, 122.0, 121.7, 121.1, 109.4, 108.1, 101.5, 62.9, 62.2, 52.2, 49.4, 33.1, 32.7, 25.7, 25.1, 21.4. EM-ESi m / z 694 ([M+HJ+).Example 14: synthesis of 2-(N-(2-(cyclohexylam o)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guamdino)benzamido)acet!c acid (1F-184)*NBoc 1 KOH / MeOH NH1F-139 1F-184

[0107] “Compound 1F-184 was obtained by the specific procedure described below:

[0108] Intermediate 1F-139 obtained for the synthesis of 1F-181 of Example 12 (32 mg; 0.05 mmol) was treated with 5% KOH in methanol (0.75 mL) a 50 °C for 45 min. The reaction mixture was diluted with H2O (1.5 mL), adjusted to pH 2-3 with 5% HCi and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with NaCI saturated solution (1 x 5 mL), dried (Na2SO4) and the solvent was evaporated to provide 26 mg of a product that was treated with TFA-CH2CI2 solution (1:1; 0.65 mL) overnight at room temperature and the solvent was evaporated. The residue was purified by flash chromatography (silica gel) eluted with CH2Cl2-MeOH gradient to provide 15 mg (61%) of 1 F-184 as a white solid.

[0109] Rf0.40 (CH2CI2-MeOH; 9:1). Mp 206-207 °C.1H NMR (300 MHz; CDCh / MeOD): 5 7.46-7.26 (m, 4H), 6.98 (maj) and 6.95 (min) (rot., s, 1H), 6, 87 (mln) and 6.86 (maj) (rot., s, 2H), 4.06 (maj) and 3.87 (min) (rot., s, 2H), 3.99 (min) and 3.90 (maj) (rot., s, 2H), 3.59 (maj) and 3.48 (min) (rot., m, 1H), 2.30 (s, 6H), 1.76-1.67 (m, 2H), 1.53-1.45 (m, 3H), 1.26-1.10 (m, 2H), 1.02-0.88 (m, 2H), 0.86- 0.76 (m, 1H).13C NMR (75 MHz, CDCh / MeOD): 6175.9 (maj) and 175.8 (min) (rot.), 172.1 (maj) and 171.3 (min) (rot., C-12), 168.9 (min) and 168.2 (maj) (rot.), 154.8, 140.2 (maj) and 140.1 (min) (rot.), 137.2 (min) and 137.1 (maj) (rot.), 135.5 (min) and 135.1 (maj) (rot.), 134.3, 130.2, 129.7 (maj) and 129.5 (min) (rot.), 126.9 (min) and 126.4 (maj) (rot.), 125.4, 123.4 (C-3), 123.0 (C-7), 56.2 (maj) and 56.0 (min) (rot., C-19), 54.0 (maj) and 53.8 (min) (rot.), 49.1 (min) and 48.4 (maj) (rot.), 32.6 (maj) and 32.4 (min) (rot.), 25.3, 25.1 (min) and 24.9 (maj) (rot.), 21.2. EM-ESI m / z 480 ([M+H]+).Example 15: synthesis of N-(1-(benzo[d][1,3]dioxol-5-yl)-2-(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5-dimethylphenyl)guanidino)benzamide (1F-188)1F-188

[0110] Following general procedure A, piperonaldehyde (20 mg; 0.13 mmol), 5- aminomethyl indole (19 mg; 0.13 mmol), 1F-123 (50 mg; 0.13 mmol) and cyclohexyl isonitrile (17 pL; 0.13 mmol) in 0.5 mL of MeOH were reacted for 5 days. The reaction was purified by column chromatography (silica gel) eluted with hexane-EtOAc to provide 53 mg of a product that was treated with 1.2 mL of TFA-CH2Cl2(1:1) for 4 h at room temperature, and then the pH was adjusted to 8 by the addition of NaHCO3saturated solution at 0 °C. The mixture was extracted with CH2Cl2(5 x 2,5mL), the combined organic layers were dried (Na2SO4) and the solvent was evaporated. The residue was purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH to provide 26 mg (39%, total) of 1 F-188 as a white solid.

[0111] Rf0.38 (CH2Cl2-MeOH, 9:1). Mp 137 °C.1H NMR (300 MHz, CDCI3): 68.20 (bs, 1H), 7.65 (s, 1H), 7.56 (dist t, 1H), 7.37-7.33 (m, 2H), 6.92 (s, 1H), 6.88 (s, 1H), 6.86-6.83 (m, 3H), 6.71 (d, J = 7.9 Hz, 1H), 6.26 (d, J = 7.4 Hz, 1H), 5.91 (s, 2H), 5.43 (d, J = 5.8 Hz, 1H), 3.66-3.56 (m, 1H), 2.28 (s, 6H), 1.83-1.55 (m), 1.30-0.96 (m).13C NMR (75 MHz, CDCI3): 5 169.3 (C-14), 165.9, 154.6 (C-5), 148.2, 147.8, 140.3, 135.5, 135.4, 134.2, 131.6, 130.2, 129.8, 128.5, 126.2, 124.1, 123.1, 121.4, 108.6, 107.8, 101.4 57.5, 49.2, 32.7, 32.6, 25.5, 24.9, 24.8, 21.3. EM-ESI m / z 542 ([M+H]⁺).Scheme of the synthetic route for guanidine aipha-aminoacyl amide compounds of the present invention where A=N.H2N2 Pd(C), H21G-125, 1G-126HN'CN1G-129HCI / EtOH1G-130, 1G-132General procedure B for the synthesis of intermediates of guanidine alpha-aminoacyl amide compounds where A~N by Ugi reaction followed by catalytic hydrogenation

[0112] To a solution of 3-nitrobenzoic acid (1 eq.), an aldehyde (1 eq.), an amine (1 eq.) in MeOH (2 mL / mmol) an isonitryl (1 eq.) was added and the solution was reacted for 2 days at room temperature. The reaction was quenched by evaporation of the solvent or by addition of 5% HCI, followed by extraction with CH2CI2(3x). The combined organic layers were dried (Na2SO4) and the solvent was evaporated. The resulting residue was purified by flash column chromatography or washing, and it was dissolved in EtOAc (20 mL / mmol). The solution was subjected to heterogeneous catalytic hydrogenation using 10% palladium on activated charcoal at room temperature. Upon completion of the reaction (1 h to 48 h), the reaction mixture was filtered through a Celite bed, and the solvent was evaporated in rotavap. The resulting crude was purified by silica gel column chromatography to provide 1G- 125 and 1G-126 as solids.Example 16: synthesis of intermediate 3-amino-N-(1~(2,5~dichlorophenyl)-2~ oxo-2-(p-tolylammo)ethyl)-N-methylbenzamide (1G- 25)

[0113] According to General procedure B, to a solution of 3-nitrobenzoic acid (90 mg; 0.54 mmol), 2,5-dichlorobenzaldehyde (89 mg; 0.51 mmol), methylamine hydrochloride (37 mg; 0.55 mmol) and triethylamine (75 pL; 0.54 mmol) in 1 mL of MeOH, 4-methylphenyl isonitrile (59 mg; 0.50 mmol) was added and the solution was reacted for 2 days at room temperature. Then 5% HCI (6 mL) was added and the mixture was extracted with CH2CI2(3 x 5 mL). The combined organic layers were dried (Na2SO4) and the solvent was evaporated. The residue was washed with a mixture of hexane-CH2CI2(2:1) to provide 160 mg of product, which was dissolved in 7 mL of EtOAc and treated with hydrogen at 1 bar in the presence of 10% palladium on carbon (20 mg) for 1 h at room temperature. The mixture was filtered through a Celite bed and the solvent was evaporated. The residue was purified by flash chromatography (silica gel) eluted with CH2Cl2-EtOAc gradient to provide 90 mg (60%) of 1G-125 as a white solid.Example 17: synthesis of intermediate 3-amino-N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)benzamide (1G-126)HCl1G-126

[0114] To a solution of 3-nitrobenzoic acid (88 mg; 0.53 mmol), 37% formaldehyde (40 pL; 0.49 mmol) and 4-chloroaniline (69 mg; 0.54 mmol) in 1 mL of MeOH, 3,4-dimethoxyphenethyl isonitrile (103 mg; 0.54 mmol) was added and the solution wasreacted for 2 days at room temperature. Then the solvent was evaporated and the residue was purified by flash chromatography (silica gel) eluted with CH2Cl2-EtOAc gradient to provide 182 mg of product, which was dissolved in 7 mL of EtOAc and treated with hydrogen at 1 bar in the presence of 10% Pd on carbon (20 mg) for 1 h at room temperature. The mixture was filtered through a Celite bed and the solvent was evaporated. The residue was purified by flash chromatography (silica gel) eluted with gradient CH2CI2-EtOAc to provide 120 mg (70%) of 1G-126 as a white solid.

[0115] Rf 0.23 (CH2CI2-EtOAc, 1:1). Mp 82-83 °C.1H NMR (300 MHz, MeOD): 6 7.22 (d, J = 8.6 Hz, 2H), 7.09 (d, J = 8.6 Hz, 2H), 6.93 (t, J = 7.8 Hz, 1H), 6.86 (d, J = 1.6 Hz, 1 H), 6.85 (d, J = 8.0 Hz, 1 H), 6.76 (dd, J1 = 8.0 Hz, J2 = 1.4 Hz, 1 H), 6.71 (s, 1H), 6.64 (d, J = 7.8 Hz, 1H), 6.54 (d, J = 7.5 Hz, 1H), 4.44 (s, 1H), 3.80 (s, 3H), 3.79 (s, 3H), 3.44 (t, J = 7.0 Hz, 2H), 2.74 (t, J = 7.0 Hz, 2H).13C NMR (75 MHz, MeOD): 5 173.7, 170.4, 150.4, 149.0, 143.9, 137.2, 133.6, 133.3, 130.1, 130.0, 129.7, 122.2, 118.8, 117.8, 116.0, 113.6, 113.0, 56.4, 54.3, 42.1, 36.0.Example 18: synthesis of intermediate N-(4.6-dimethyipyridin~2-yi)cyanamide (1G-129):

[0116] To a solution of N-(4,6-dimethyl-2-pyridyl)thiourea (860 mg; 4.74 mmol) in 5 mL of H2O at 95 °C, a KOH solution (2.54 g, 45.23 mmol) in 6 mL of H2O was added at 95 °C and a 1,5 M lead acetate solution in water (3.5 mL, 5.25 mmol) was immediately added (according to US20180244608, incorporated herein by reference). The reaction mixture was kept at 95 °C for 30 min and a black precipitate was formed. The precipitate was filtered with vacuum through filter paper and the mother liquor was adjusted to pH 7 with glacial AcOH. The formed solid was filtered, washed with iced H2O and dried to provide 690 mg (99%) of 1G129 as a white solid

[0117] Rf 0.32 (CH2CI2-MeOH, 95:5); mp 222 °C.1H NMR (300 MHz, MeOD): 5 6.78 (s, 1H), 6.42 (s, 1H), 2.34 (s, 3H), 2.30 (s, 3H).13C NMR (75 MHz, MeOD): 6 162.7, 157.2, 147.8, 114.3, 114.1, 110.2, 21.6, 18.7.Genera / procedure C for the synthesis of guanidine alpha-aminoacyl amide compounds of the present invention where A~N, by the coupling of 1G-125 and / or 1G-126 with cyanamide (1G-129)

[0118] The compounds 1G-125 or 1G-126 (1 equiv) and 1G-129 (1.9 to 2.6 equiv) were dissolved in absolute EtOH (7 mL / mmol) and were treated with a solution of 15.5% HCI in IPA (1.5 equiv). The mixture was refluxed for 2 or 3 days. The reaction mixture was allowed to reach room temperature, and was poured over water (100 to 166 mL / mmol). The pH was adjusted to 8-9 using 20% NaOH. The mixture was extracted with CH2CI2, the combined organic layers were dried (Na2SO4) and evaporated under reduced pressure. The residue was purified by column chromatography (silica gel) to provide the final products 1G-132 and 1G-130.

[0119] Exemplary syntheses of the compounds derived from guanidine alpha¬ aminoacyl amide where A is N are described below:Example 19: synthesis of N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)-N-methylbenzamide (1G-132)CK, NHNHCl1G-132

[0120] According to General procedure C, to a solution of 1G-125 (65 mg, 0.15 mmol) and 1G-129 (58 mg, 0.39 mmol) in 1 mL of absolute EtOH, 15.5% HCI in isopropanol (60 pL; 0.21 mmol) was added and the solution was refluxed for 3 days. Then it was poured over H2O (25 mL) and adjusted to pH 8-9 with 20% NaOH. The mixture was extracted with CH2CI2(5 x 20 mL), the combined organic layers were dried (Na2SO4) and the solvent was evaporated. The residue was purified by columnchromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide 21 mg (24%) of 1G-132 as a white solid.

[0121] Rf 0.39 (CH2CI2-MeOH-NH3, 9:1:0,05). Mp 226 °C.1H NMR (300 MHz, CDCI3 / MeOD): 07.67 (d, J = 1.2 Hz, 1H), 7.47 (d, J = 8.1 Hz, 2H), 7.37 (d, J = 8.4 Hz, 1H), 7.33 (t sup., 1H), 7.30 (d, J = 7.5 Hz, 1H), 7.17-7.13 (m, 3H), 7.09 (d, J = 8.7 Hz, 2H), 6.58 (min) and 6.53 (maj) (rot., s, 1H), 6.54 (maj) and 5.77 (min) (rot., s, 1H), 6.38 (maj) and 6.23 (min) (rot., s, 1H), 2.93 (min) and 2.85 (maj) (rot., s, 3H), 2.36 (s, 3H), 2.29 (s.3H), 2.16 (s, 3H).13C NMR (75 MHz, CDCI3 / MeOD): 5 172.4, 167.1, 155.0, 154.9, 152.2, 149.8, 145.8, 136.6, 135.4, 134.4, 134.1, 133.7, 133.3, 131.2, 130.8, 130.2, 129.9, 129.5, 125.8, 122.4, 120.1, 117.5, 110.8, 64.2 (min) and 59.4 (maj) (rot.), 35.7-31.5, 24.0, 21.0. EM-ESI m / z 588,9 ([M+H]+)Example 20: synthesis of N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)benzamide (1G-130)Cl1G-130

[0122] According to General procedure C, to a solution of 1G-126 (65 mg, 0.14 mmol) and 1G-129 (40 mg, 0.27 mmol) in 1 mL of absolute EtOH, 15.5% HCI in isopropanol (60 pL; 0.21 mmol) was added and the solution was refluxed for 2 days. Then it was poured over H2O (15 mL) and adjusted to pH 8-9 with 20% NaOH. The mixture was extracted with CH2Cl2(6 x 10 mL), the combined organic layers were dried (Na2SO4) and the solvent was evaporated. The residue was purified by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide 23 mg (27%) of 1G-130 as a white solid.

[0123] Rf 0.35 (CH2Cl2-MeOH-NH3, 9:1:0.1). Mp 129 °C.1H NMR (300 MHz, CDCl3): 67.23 (t, J = 7.7 Hz, 1 H), 7.15 (d, J = 8.7 Hz, 2H), 7.09 (d, J = 7.7 Hz, 1H), 7.01 (d, J = 7.5 Hz, 1H), 6.93 (s, 1H), 6.92 (d, J = 8.6 Hz, 2H), 6.77 (d, J = 8.5 Hz,1 H), 6.73 (s, 1H), 6.72 (d, J = 7.6 Hz), 6.61 (dist. t, 1H), 6.51 (s, 1H), 6.16 (s, 1H), 4.43 (s, 1 H), 3.85 (s, 3H), 3.79 (s, 3H), 3.54 (q, J = 5.9 Hz, 2H), 2.77 (t, J = 6.9 Hz, 2H), 2.37 (s, 3H), 2.07 (s, 3H).13C NMR (75 MHz, CDCI3): 6 170.8, 168.5, 154.8, 154.5, 152.5, 149.9, 149.1, 147.7, 145.9, 142.4, 135.9, 132.8, 131.2, 129.8, 129.5, 128.5, 126.7, 124.3, 124.0, 120.8, 117.5, 111.9, 111.4, 110.6, 55.9, 54.8, 40.6, 35.1, 24.1, 20.9. EM-ESI m / z 614,8 ([M+H]+).Example 21: Anti-proliferative potential of the novel family of RAC1 inhibitors. Cell culture and viability assays

[0124] Cell lines were confirmed to be mycoplasma-free using the e-Myco kit (Boca Scientific). All cell lines were kept in DMEM medium supplemented with 2 pM glutamine, 100 U / mL penicillin, 100 mg / mL streptomycin and heat-inactivated 10% fetal bovine serum (FBS). For cell viability assays, the cells were seeded at 500- 2000 cells / well in 96-well plates and were treated the following day with increasing doses of the compounds for 3 days. Cell viability was evaluated by standard MTT assays using reagents from Sigma according to the manufacturer’s protocols and measuring the absorbance at 570 nm with a Thermo Multiskan FC plate reader. The data were normalized to untreated controls (100% viability). Each combination of cell line and compound was tested in 1-3 independent assays, each with 4 replicates. The dose response curves were plotted using a non-linear regression model and IC50 values were determined from the fitted curves using GraphPad Prism. The average IC50 derived from 1-3 independent assays, each with 4 replicates, is reported.

[0125] Firstly, the potential of 14 compounds (50 M) to inhibit the interaction between RAC1 and its GEF (TIAM1) and proliferation in colorectal carcinoma (HT29) and lung cancer (A549) was tested by an alpha assay and a standard MTT assay, respectively. This initial evaluation showed that the compounds 1F-182, 1F- 188, 1F-189, 1G-55, 1G-56, 1G-57 and 1G-58 are able to reduce both the RAC1- TIAM1 interaction and the proliferation of cancer cells (Figure 2).Figure 2 shows the dose-response curve of the compounds on a hepatocellular carcinoma cell line, from which the IC50 values listed in Table 2 below for 4 cancercell lines were calculated: hepatocellular carcinoma (HepG2), colorectal carcinoma (HT29), lung cancer (A549) and breast cancer (A375) in vitro cell lines.

[0126] Table 4. Half-maximal inhibitory concentration (IC50) of the HIT compounds in cancer cell lines determined by MTT assay.IC50Cell lineCompound HepG2 HT29 A549 A3751F-182 8.7 ± 0.1 6.0 ± 0.1 16.7 ± 0.5 7.3 ± 0.31F-189 21.0 ± 0.3 4.7 ± 0.2 >50 17.1 ± 0.41G-56 6.7 ± 0.1 3.1 ± 0.1 7.3 ± 0.2 5.0 ± 0.11G-55 3.5 ± 0.1 2.9 ± 0.4 3.9 ± 0.1 2.7 + 0.11F-188 13.8 ± 0.1 6.8 ± 0.1 28.4 ± 0.7 12.7± 0.11G-58 25.5 ± 0.2 13.8 ± 0.3 >50 26.8 ± 0.11G-57 13.1 ± 0.1 8.8 ± 0.3 32.2 ± 0.9 12.8 ± 0.1

[0127] Compounds 1 G-55 and 1 G-56 exhibit a higher anti-proliferative effect on the cell lines according to the in vitro assays.Example 22: Antitumor effect of the compounds in an animal modelin vivo experiments

[0128] Animal care and experimental procedures were carried out according to institutional guidelines, and complied with state authority regulations for animaltesting. In all assays, body weight and general health were monitored every two days.Animal model of orthotopic hepatocellular carcinoma

[0129] Six- to eight-week-old male C3H / HeJ mice were used. Fibrosis was induced by intraperitoneal (i.p.) injections of thioacetamide (TAA) (one 200 mg / kg dose; Sigma-Aldrich, St. Louis) 3 times per week for 42 days. On day 30, orthotopic tumors were established by subcapsular inoculation of 1.25 x 105Hepa129 cells in the left liver lobe by laparotomy. Seven days after tumor implantation, a group of mice was administered 1G-55 (19 mg / kg) or control by intraperitoneal injection in an aqueous suspension of 12.5% Cremophor EL, 12.5% DMSO, for 2 weeks, 3 times per week. Twelve days after the initiation of the treatment, the mice were euthanized and the tumor volume (mm3) was calculated using the formula TT / 6 X longer diameter x (shorter diameter)2, measured with a caliper.Animal model of colorectal carcinoma

[0130] Six- to eight-week-old male BalbC mice were used. Tumors were established by subcutaneous inoculation of 5 x 105CT-26 cells into the left flank. Ten days after tumor implantation, a group of mice was administered 1G-55 (19 mg / kg) or control by intraperitoneal injection in an aqueous suspension of 12.5% Cremophor EL, 12.5% DMSO, for 2 weeks, 3 times per week. When tumors reached 3000 mm3, the mice were euthanized. The tumor volume was calculated as described above.

[0131] In order to evaluate if the compounds of the invention also have antitumor activity in vivo, the therapeutic potential of 1G-55 was tested in mouse models of orthotopic hepatocellular carcinoma (HCC) (Figure 3A) and subcutaneous colorectal carcinoma (CCR) (Figure 3B). Notably, 1G-55 reduced tumor growth in both models (Figure 3). In addition, in the HCC model, the treatment with 1G-55 reduced intrahepatic metastasis and the development of ascites (Figure 3A).Example 23: Anti-fibrotic effect of the compounds in an animal model of liver injuryMouse model of liver fibrosis

[0132] Six- to eight-week-old male BalbC mice were used. Fibrosis was induced by intraperitoneal (i.p.) administration of 200 mg / kg of body weight of thioacetamide (TAA) (Sigma-Aldrich, Missouri, USA) 3 times per week for 6 weeks. Then, a group of mice was administered 1G-55 (19 mg / kg) or control by intraperitoneal injection in an aqueous suspension of 12.5% Cremophor EL, 12.5% DMSO for 4 weeks, 3 times per week. During the administration of the treatment, TAA was administered 2 times per week (200 mg / kg). On week 10, the mice were euthanized and liver tissue was fixed in 10% formaldehyde. Then, the liver samples were embedded in paraffin and 5 pm sections were stained with Sirius Red for fibrillar collagen detection. A computer-assisted morphometric analysis was carried for the quantitative analysis of the Sirius Red-stained area. About 50 images per specimen were captured and analyzed with an optical microscope (100X magnification) using the color threshold detection system from ImageJ software (NIH, USA). The results are expressed as positive area percent.Serum biochemical marker detection

[0133] On completion of the in vivo FHA experiments, the mice were euthanized and blood samples were collected for serum quantification. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were quantified using an Aiinity instrument (Abbott).Statistical analysis

[0134] The data were expressed as mean ± SEM unless otherwise stated. The statistical analysis was carried out using the GraphPad Prism software. Paired and unpaired t-test, Mann-Whitney test and two-way analysis of variance were applied. The data distributions were analyzed by Shapiro-Wilk test. A value of P <0.05 was considered statistically significant.Results

[0135] Prior reports disclose that the RAC1 pathway is involved in fibrosis pathogenesis. Once activated, RAC1 induces cell migration, proliferation, inflammation and ROS production in different cell types, such as hepatic stellate cells (HSCs) and hepatic macrophages. In order to assess the hypothesis that pharmacological inhibition of RAC1 by 1 G-55 may reduce liver fibrosis, the presence of collagen deposits in liver sections of mice with fibrosis induced by 6 weeks of thioacetamide administration, treated or untreated with 1G-55 (Figure 4), was determined. Notably, 1 G-55 markedly reduced collagen deposits in the livers of mice with fibrosis (Figure 4).

[0136] Then, the effects of 1G-55 in cell viability and the phenotype of hepatic stellate cells (HSCs), which play a key role in the production of type I collagen during the fibrogenic process, were assessed. As shown in Figure 5A, 1G-55 markedly reduced the survival of HSCs in vitro with an IC50 similar to the value observed in cancer cells. In addition, 1G-55 reduces the expression of pro-fibrogenic markers CollA2 and u-SMA (Figure 5B) in HSCs.Example 24: liver injury reduction in mouse models of acute and acute-on- chronic liver failure by compound 1G-55.Mouse model of acute liver failure (ALF) with Concanavalin-A

[0137] Six- to eight-week-old male C57BL / 6J mice that were subjected to overnight fasting were treated with Con-A 30 mg / kg of body weight diluted in saline solution (0.9% NaCI) by single injection into the tail vein. Treatment with 1G-55 at 19 mg / kg was administered 30 minutes after Con-A injection, and the mice were euthanized 6 hours later.

[0133] Mouse model of acute liver failure (ALF) with acetaminophen

[0139] APAP (Sigma-Aldrich) was dissolved in hot PBS at 70 °C, and the solution was incubated in a water bath at 55 °C for 30 minutes and injected by intraperitoneal route in six- to eight-week-old male C57BL / 6J mice that were subjected to overnight fasting (600 mg / kg). One hour after APAP administration, a group of mice wasadministered 1G-55 at 19 mg / kg or control by intraperitoneal route. All animals were euthanized 6 hours later.

[0140] Model of acute-on-chronic liver failure

[0141] Fibrosis was induced by the intraperitoneal (i.p.) administration of 0.2 mg / g of body weight of thioacetamide (TAA) (Sigma-Aldrich, Missouri, USA) 3 times per week for 10 weeks. The last dose of TAA was 0.4 mg / g of body weight, and 24 hours later APAP (600 mg / kg) and LPS (25 pg / kg) were injected by intraperitoneal route. Blood samples were collected after 24 hours and the mice were euthanized after 11 days.Results:

[0142] Prior reports indicate that RAC1 is involved in the production of ROS and in the inflammatory process. In consequence, the inventors posed the hypothesis that 1G-55 may have a therapeutic effect 1G-55 in acute liver failure (ALF) induced by oxidative damage or immune system-mediated acute hepatitis. In order to confirm this hypothesis, the protective effect of 1 G-55 was assessed in two ALF models. The first ALF model was induced with acetaminophen (APAP), where ROS play a major role, and the second one with Concanavalin A (ConA) to mimic immune system-mediated acute hepatitis, mediated by macrophages, T cells, natural killer cells and neutrophils (Figure 6A). Notably, the treatment with 1G-55 significantly reduced the degree of liver injury, as evaluated by AST and ALT measurements in both ALF models (Figure 6B).

[0143] In addition, the inventors evaluated the protective effect of 1 G-55 in a mouse model of acute-on-chronic liver failure (ACLF) that involved establishment of liver fibrosis by administering TAA for 10 weeks, followed by the induction of acute liver injury by the administration of acetaminophen (APAP) and lipopolysaccharide (LPS) to generate both oxidative stress and inflammation, which are the specific features of ACLF. In line with the results obtained in ALF models, the treatment with 1G-55 resulted In significantly lower levels of alanine aminotransferase (ALT), indicating a reduction in liver injury (Figure 7).

[0144] Finally, the effects of 1 G-55 in liver explants from patients (healthy livers and acute liver failure patient livers) were evaluated (Figure 8). The in vitro incubation of liver explants with 1G-55 induced no changes in transaminase (ALT) levels (A).Then, a heathy liver exp ant was exposed to Concanavalin A to induce liver injury. As shown in (B), the treatment with 1G-55 enhances ALT levels. Finally, the incubation of acute liver failure patient liver explants induced a reduction in ALT released into the supernatant (C).ReferencesBernal, W., et al,, Acute liver failure: A curable disease by 2024? J Hepatol, 2015. 62(1 Suppl): p, S112-20.Wu, Z., et al., Acute liver failure: mechanisms of immune-mediated liver injury. Liver Int, 2010. 30(6): p. 782-94.Yan, M., et al., Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions. Redox Biol, 2018. 17: p. 274-283.Mossanen, J. C, and F. Tacke, Acetaminophen-induced acute liver injury in mice. Lab Anim, 20 5, 49(1 Suppl): p. 30-6,Dong, V., R. Nanchal, and C. J. Karvellas, Pathophysiology of Acute Liver Failure. Nutr Clin Pract, 2020. 35(1): p. 24-29.Walayat, S., et al., Role of N-acetylcysteine in non-acetaminophen-related acute liver failure: an updated meta-analysis and systematic review. Ann Gastroenterol, 2021. 34(2): p. 235-240.Rovegno, M., et al., Current concepts in acute liver failure. Ann Hepatol, 2019. 18(4): p, 543-552.Thanapirom, K., et al., The incidence, etiologies, outcomes, and predictors of mortality of acute liver failure in Thailand: a population-base study. BMC Gastroenterol, 2019. 19(1): p.18.Wang, T., et al., RHO GTPase family in hepatocellular carcinoma. Exp Hematol Oncol, 2022. 11(1): p. 91.Sauzeau, V,, J. Beignet, and C. Bailly, Rad as a Target to Treat Dysfunctions and Cancer of the Bladder. Biomedicines, 2022. 10(6).Bianchi-Smiraglia, A., et al., Author Correction: Regulation of local GTP availability controls RAC1 activity and cell invasion. Nat Common, 2021. 12(1): p. 6482.Marel, H. and A. Malliri, GEFs: Dual regulation ofRad signaling. Small GTPases, 2017.8(2): p. 90-99,Raz, L., et al., Role ofRad GTPase in NADPH oxidase activation and cognitive impairment following cerebral ischemia in the rat. PLoS One, 2010. 5(9): p. e12606.1 Hordijk, P. L., Regulation of NADPH oxidases: the role of Rac proteins. Circ Res, 2006. @8(4): p. 453-62.Choi, S. S., et al., Sustained activation of Rac1 in hepatic steiiate cells promotes liver injury and fibrosis in mice. Hepatology, 2006. 44(5): p. 1267-77.Nagase, M., et al., Deletion ofRadGTPase in the Myeloid Lineage Protects against Inflammation-Mediated Kidney Injury in Mice. PLoS One, 2016. 11(3): p. e0150886.Jiang, J. X,, et al., Rad signaling regulates cigarette smoke-induced inflammation in the lung via the Erk1 / 2 MAPK and ST AT3 pathways. Biochim Biophys Acta Mol Basis Dis, 2017. 1863(7): p. 1778-1788.Pohlmann, S., et al., Hepatic Rad GTPase contributes to liver-mediated basal immune homeostasis and LPS-induced endotoxemia. Biochim Biophys Acta Mol Cell Res, 2018.1865(9): p. 1277-1292.Hammier, J. K., et al., Inhibition of Rad Signaling Downregulates Infiammasome Activation and Attenuates Lung Injury in Neonatal Rats Exposed to Hyperoxia. Neonatology, 2017.111(3): p. 280-288.Eitel, J., et al., Rad regulates the NLRP3 inflammasome which mediates IL-1 beta production in Chlamydophila pneumoniae infected human mononuclear cells. PLoS One, 2012. 7(1): p. 630379.Bopp, A., et al., Rad modulates acute and subacute genotoxin-induced hepatic stress responses, fibrosis and liver aging. Cell Death Dis, 2013. 4(3): p. e558.Hwaiz, R., et al., Rad -dependent secretion of platelet-derived CCL5 regulates neutrophil recruitment via activation of alveolar macrophages in septic lung injury. s Leukoc Biol, 2015. 97(5): p. 975-984.Yu, C., et al., Rad signaling regulates neutrophil-dependent tissue damage in experimental colitis. Eur J Pharmacol, 2014. 741: p. 90-6.Bayo, J,, et a I,, Bioinformatic analysis of RHO family of G TPases identifies RAC1 pharmacological inhibition as a new therapeutic strategy for hepatocellular carcinoma. Gut, 2021. 70(7): p. 1362-1374.Shi, Y., et al., Rad-Mediated DNA Damage and Inflammation Promote Nf2 Tumorigenesis but Also Limit Cell-Cycle Progression. Dev Cell, 2016. 39(4): p. 452-465.Singh, A., et al., NADPH oxidase 4 modulates hepatic responses to lipopolysaccharide mediated by Toll-like receptor-4. Scl Rep, 2017. 7(1): p. 14346.Murillo, M. M., et al., Activation of NADPH oxidase by transforming growth factor-beta in hepatocytes mediates up-regulation of epidermal growth factor receptor ligands through a nuclear factor-kappaB-dependent mechanism. Biochem J, 2007. 485(2): p. 251-9.Maitra, U., et al., IRAK-1 contributes to lipopolysaccharide-induced reactive oxygen species generation in macrophages by inducing NOX-1 transcription and Rad activation and suppressing the expression of antioxidative enzymes. J Biol Chem, 2009. 284(51): p.35403-11.Koga, H., et al., Tetratricopeptide repeat (TPR) motifs of p67(phox) participate in interaction with the small GTPase Rac and activation of the phagocyte NADPH oxidase. J Biol Chem, 1999. 274(35): p. 25051-60.Diekmann, D., et al., interaction of Rac with p67phox and regulation of phagocytic NADPH oxidase activity. Science, 1994. 265(5171): p. 531-3.Zhao, X., K. A. Carnevale, and M. K. Cathcart, Human monocytes use Rac1, not Rac2. in the NADPH oxidase complex. J Biol Chem, 2003. 278(42): p. 40788-92.Cheng, P. W., et al., Resveratrol Inhibition of Rad -Derived Reactive Oxygen Species by AMPK Decreases Blood Pressure in a Fructose-Induced Rat Model of Hypertension. Sci Rep. 2016.6: p. 25342.Zimmer, S., et al., Inhibition of Rad GTPase Decreases Vascular Oxidative Stress, improves Endothelial Function, and Attenuates Atherosclerosis Development in Mice. Front Cardiovasc Med, 2021. 8: p. 680775.Pan, Y., et al., Inhibition of Rad ameliorates neuronal oxidative stress damage via reducing Bcl-2 / Rad complex formation in mitochondria through PI3K / Akt / mTOR pathway. Exp Neurol, 2018. 380: p. 149-166.O’Donovan, D. H.; Rozas, I. A Concise Synthesis of Asymmetrical N, N’-Disubstituted Guanidines. Tetrahedron Lett. 2011, 52 (32), 4117-4119.Riches, A. G.; Cablewski, T.; Glattauer, V.; Thissen, H.; Meagher, L. Scalable synthesis of an integrin-binding peptide mimetic for biomedical applications. Tetrahedron 68 (2012) 9448-9455).(Brian R. Linton, f Andrew J. Carr,; Brendan P. Omer, and Andrew D. Hamilton. A versatile one-pot synthesis of 1,3-Substituted Guanidines from Carbamoyl Isothiocyanates. J. Org. Chem. 2000, 65, 1566-1568)

Claims

ClaimsHaving described and represented the nature and scope of the invention and the manner in which it shouid be put into practice, the inventors claim the following for exclusive right and protection:

1. A compound of general formula (i):or a pharmaceutically acceptable salt thereof, characterized in that:- A is independently selected from the group consisting of CH or N;R1 is selected from the group consisting of: H; (C1-C4) alkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Brand I; mono- or dihydroxy aryl; 6-membered heteroaryl containing 1 heteroatom selected from the group consisting of N, O, and S; methyl benzoate; methyl acetate; S-methyl imidazole propanoate;R2 is selected from the group consisting of: H (hydrogen), substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Br and I; mono- or dimethoxy phenethyl, benzo[1,3]dioxolyl, N-indolyl methyl;R3 is selected from the group consisting of: H; (C1-C4) alkyl; (C5- C6) cycloalkyl; substituted or unsubstituted aryl; mono- or disubstituted aryl halide, where the halides are selected from the group consisting of Cl, F, Brand I; tert-butyl (disubstituted propyl), mono- or dimethoxy phenethyl, N-indolyl ethyl.

2. The compound according to claim 1 characterized in that A is selected from the group consisting of CH and N R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4-chlorophenyi; 4-fluorophenyl; 3,5- dimethylphenyi; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyi; 2,5-dichlorophenyl; 1 -methyl- 1 H-indol- 3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1H-indol-3- yl)ethyl; 4-bromo-2-methylphenyl; tert-butyl; p-tolyl.

3. The compound according to claim 1 characterized in that: if A is -CH, then R1 is selected from the group consisting of: methyl; 4-hydroxyphenyl; 4- chlorophenyi; 4-fluorophenyl; 3,5-dimethylphenyl; 2-methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2- chlorophenyl; 2,5-dichlorophenyl; 1-methyl-1H-indol-3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1H-indol-3- yl)ethyl; 4-bromo-2-methylphenyl; tert-butyl; p-tolyl.

4. The compound according to claim 1 characterized in that: if A is -N-, then R1 is selected from the group consisting of: methyl; 4-chlorophenyl; R2 is selected from the group consisting of: H, 2,5-dichlorophenyl; R3 is selected from the group consisting of: p-tolyl; 3,4-dimethoxy phenethyl.

5. The compound according to claims 1 to 4, characterized in that the compound is selected from the group consisting of:- Methyl 2-(N-(2-(cyclohexylamino)-2-oxoethyl)-3-(3-(3,5- dimethylphenyl)guanidino)benzamido)acetateN-(2-((4-bromo-2-methylphenyl)amino)-2-oxo-1-phenylethyl)-3-(3- (3,5-dimethylphenyl)guanidino)-N-(4-hydroxyphenyl)benzamide N-(2-(t-butylamino)-2-oxo-1-phenylethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-(pyridin-3-yl)benzamide2-(N"(2-(cyclohexylamino)-2-oxoethy)"3-(3-(3,5- dimethylphenyl)guanidino)benzamido)acetic acidN-(1 -(benzo[d][1,3]dioxol-5-yl)-2-(cyclohexylamino)-2-oxoethyl)-3- (3-(3,5-dimethyiphenyl)guanidino)benzamidemethyl 2-(3-(3-(3,5-dimethylphenyl)guanidino)-N-(2-((4- methoxyphenethyl)amino)"2-oxoethyl)benzamido)benzoate - methyl 2-(N-(1-(benzo[d][1,3]dioxoi-5-yl)-2-(cyciohexylamino)-2- oxoethyl)-3-(3-(3,5-dimethylphenyl)guanidino)benzamido)-3-(1H- imidazol-5-yl)propanoateN-(2-((2-(1H-indol-3-yl)ethyl)amino)-1-(1-methyl-1H-indol-3-yl)-2- oxoethyl)-N"(3,5-dimethylphenyl)-3-(3"(3,5- dimethylphenyl)guanidino)benzamideN-(1-(2,5-dichlorophenyl)-2-oxo-2"(p-tolylamino)ethyl)-3-(3”(3,5" dimethylphenyi)guanidino)-N-methyibenzamideN-(1-(2-chlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(3,5- dimethylphenyl)guanidino)-N-methylbenzamideN-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2- oxoethyl)-3-(3-(3,5-dimethylphenyl)guanidino)benzamide - N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)-3-(3-(315- dimethylphenyl)guanidino)-N-(4-fluorophenyl)benzamide N-(2"((3,4-dimethQxyphenethyi)amino)-2-oxoethyl)-3"(3-(3,5- dimethylphenyl)guanidino)-N-(4-hydroxyphenyl)benzamide N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2- oxoethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)benzamide N-(1"(2,5-dichlorophenyl)-2-oxo-2-(P"tolylamino)ethyl)"3-(3-(4,6- dimethylpyridin-2-yl)guanidino)-N-methylbenzamide6. The compound according to claims 1 to 5, characterized in that said compound is for the treatment of a medical condition mediated by the Rho GTPase cell protein.

7. The compound according to claims 1 to 5, characterized in that said compound is used for the treatment of a medical condition mediated by the RAC1 cell protein.

8. The compound according to ciaims 6 or 7, characterized in that said medical condition is selected from the group consisting of chronic liver inflammation, acute liver failure, acute-on-chronic liver failure and liver fibrosis.

9. The compound according to claims 6 or 7, characterized in that said medical condition is selected from the group consisting of liver cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

10. A pharmaceutical composition characterized by comprising a compound according to claims 1 to 5, and at least one excipient, one adjuvant, or a combination thereof.

11. The pharmaceutical composition according to claim 10, characterized in that said pharmaceutical composition is used for the treatment of a medical condition featuring an increase in the activity of the Rho GTPase cell protein.

12. The pharmaceutical composition according to claim 10, characterized in that said Rho GTPase cell protein is RAC1.

13. The pharmaceutical composition according to claims 10 to 11, characterized in that said medical condition is selected from the group consisting of acute liver failure, chronic liver inflammation, acute-on-chronic liver failure and liver fibrosis1. The pharmaceutical composition according to claims 10 to 11, characterized in that said medical condition is selected from the group consisting of liver cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

15. A method for the treatment of a medical condition mediated by the RAC1 cell protein, characterized by comprising the administration to a subject in need thereof of a therapeutically effective amount of at least one compound according to claims 1 to 5 or a pharmaceutically acceptable salt, solvate, hydrate, tautomer or isomer thereof.

16. The method for the treatment of a medical condition mediated by the RAC1 cell protein according to claim 15, characterized by comprising theadministration to a subject in need thereof of a therapeutically effective amount of a pharmaceutical composition according to claim 10.

17. The method for the treatment of a medical condition mediated by the RAC1 cell protein according to claim 15, characterized in that said medical condition is selected from the group consisting of acute liver failure, chronic liver inflammation, acute-on-chronic liver failure and liver fibrosis.

18. The method for the treatment of a medical condition mediated by the RAC1 cell protein according to claim 15, characterized in that said medical condition is selected from the group consisting of liver cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, kidney cancer, spleen cancer, ovarian cancer, testicular cancer, prostate cancer, invasive oral cancer, lung cancer, breast cancer, neuroblastoma, glioma, glioblastoma, osteosarcoma, soft tissue sarcoma, leukemia, lymphoma.

19. A method for the synthesis of any of the compounds of claims 1 to 4 characterized in that: if A is -CH, said method comprises the steps of: 1) obtaining the chemical precursor N-(f-butoxycarbonyl)-N'-(3,5- dimethylphenyl)thiourea;2) from the precursor of step 1 obtaining the intermediate 3-(2-(t- butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid;3)- obtaining the guanidine alpha-aminoacyl amide compound according to claims 1 to 5 by Ugi reaction.

20. The method of claim 19 characterized in that said step 1) comprises the following sub-steps:a) preparing a solution of thiourea in dry tetrahydrofuran; b) adding sodium hydride and stirring at room temperature and cooling;c) adding di-tert-butyl dicarbonate and stirring at room temperature;d) cooling and adding a second portion of sodium hydride while stirring;e) adding trifluoroacetic anhydride dropwise, and stirring at 0°C; f) adding 3,5-dimethylaniline dropwise, and allowing to reach room temperature and stirring;g) cooling and adding water dropwise to quench the reaction;h) extracting with EtOAc;i) washing the organic layer with brine and drying; j) removing the solvents in vacuo,k) purifying by flash chromatography (silica gel) (hexane- EtOAc), andi) recrystallizing from MeOH-H2O to provide N-(f- butoxycarbonyl)-N'-(3,5-dimethylphenyl)thiourea as a white solid.

21. The method of claim 19 characterized in that said step 2 comprises the following sub-steps:a) to the solution of N-(ferf-butoxycarbonyl)-N’-(3,5- dimethylphenyl)thiourea obtained in step 1 adding benzyl 3- aminobenzoate and triethylamine in anhydrous CH2CI2; b) cooling and treating with EDCI;c) stirring under argon;d) stirring at room temperature;e) purifying the residue by flash chromatography (silica gel) (eluted with a gradient of hexane: EtOAc);f) dissolving the product obtained in step e) in EtOAc and carrying out catalytic hydrogenation with 10% palladium on activated charcoal at room temperature;g) filtering the reaction mixture through a Celite bed and evaporating the solvent at reduced pressure, to provide the desired intermediate 3-(2-(t-butoxycarbonyl)-3-(3,5- dimethylphenyl)guanidino)benzoic acid.

22. The method of claim 19 characterized in that said step 3 comprises the following sub-steps:a) to the stirring solution of 3-(2-(t-butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid obtained in step 2, adding an amine and an aldehyde in MeOH;b) adding the corresponding isonitryl;c) stirring for 2 to 7 days at room temperature;d) evaporating the solvent and purifying by column chromatography (silica gel);e) treating the product obtained in step d) with 15.5% HCI in isopropanol for 4-5 h at room temperature;f) slowly adding NaHCO3 until reaching pH 8-9;g) extracting with CH2CI2;h) drying the combined organic layers (Na2SO4) and evaporating the solvent;I) purifying by column chromatography (silica gel) to provide the compounds according to claim 1, where A = CH.

23. The method of claim 22 characterized in that said sub-steps comprise:a) adding 2,5-dichlorobenzaidehyde, methylamine hydrochloride, triethylamine, 4-methylphenyl isonitrile and 3-(2-(t- butoxycarbonyl)-3-(3,5-dimethylphenyl)guanidino)benzoic acid to MeOH;b) stirring at room temperature;c) purifying the product of step b by column chromatography (silica gel) eluted with hexane-AcOEt;d) treating with HCI in IPAe) purifying by column chromatography (silica gel) eluted with CH2Cl2-MeOH-NH3to provide N-(1-(2,5-dichlorophenyl)-2-oxo-2-(p- tolylamino)ethyl)-3-(3-(3,5-dimethylphenyl)guanidino)-N- methylbenzamide (1G-55);24. A method for the synthesis of a compound according to claim 1, characterized in that if A is N the method comprises the following steps:1) Obtaining the chemical intermediate 3-amino-N-(1-(2,5- dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-N- methylbenzamide or 3-amino-N-(4-chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2-oxoethyl)benzamide from a solution of 3-nitrobenzoic acid, an aldehyde, an amine in MeOH, adding an isonitryl by Ugi reaction followed by catalytic hydrogenation;2) Obtaining the chemical intermediate N-(4,6-dimethylpyridin-2- yl)cyanamide from a solution of N-(4,6-dimethyl-2- pyridyl)thiourea, a KOH solution, and adding a 1,5 M lead acetate solution;3) Coupling each intermediate obtained in step 1 with the intermediate of step 2, under suitable conditions, to provide N- (1-(2,5-dichlorophenyl)-2-oxo-2-(p-tolylamino)ethyl)-3-(3-(4,6- dimethylpyridin-2-yl)guanidino)-N-methylbenzamide and N-(4- chlorophenyl)-N-(2-((3,4-dimethoxyphenethyl)amino)-2- oxoethyl)-3-(3-(4,6-dimethylpyridin-2-yl)guanidino)benzamide, respectively.

25. A pharmaceutical composition for the treatment of a medical condition featuring an inflammatory process mediated by the RAC1 cell protein, according to formula (I) characterized in that:A is independently selected from the group consisting of CH or N; and if A is -CH-, R1 is selected from the group consisting of: methyl; 4- hydroxyphenyl; 4-chlorophenyl; 4-fluorophenyl; 3,5-dimethylphenyl; 2- methyl-benzoate; pyridin-3-yl; methyl acetate; (S)-methyl 3-(1H-imidazol-4-yl)-2-propanoate; R2 is selected from the group consisting of: hydrogen; phenyl; 2-chlorophenyl; 2,5-dichiorophenyl; 1-methyl-1H-indol-3-yl; benzo[d][1,3]dioxol-5-yl; R3 is selected from the group consisting of: cyclohexyl; 4-methoxyphenethyl; 3,4-dimethoxyphenethyl; 2-(1H-indol-3-yl)ethyl; 4-bromo-2-methylphenyl; tert-butyl; p-tolyl; and if A is -N-, R1 is selected from the group consisting of: methyl; 4-chlorophenyl; R2 is selected from the group consisting of: H, 2,5-dichlorophenyl; R3 is selected from the group consisting of: p-tolyl; 3,4-dimethoxy phenethyl.

26. A compound selected from the group consisting of:1G-55 1F-1821F-188Ci1G-126 1G-132and1G-130 orapharmaceutically acceptable salt thereof.

27. The compound according to claim 26, wherein the compound is1G-S528. A method for treating a liver disease to a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound selected from the group consisting of:and1F’188, or a pharmaceutically acceptable salt thereof.

29. The method according to claim 28, wherein the liver disease is selected from the group consisting of: acute liver failure, chronic liver inflammation, acute- on-chronic liver failure and liver fibrosis.

30. The method according to claim 28, wherein the compound is1G-55

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