Inhibitor of RAB30 for use in a method of treatment of metabolic dysfunction-associated steatotic liver disease (MASLD)

Abstract

Metabolic dysfunction–associated steatotic liver disease (MASLD), previously known as non- alcoholic fatty liver disease (NAFLD), is a condition characterized by the accumulation of fat in the liver of individuals with no alcohol consumption, and at least one cardiometabolic risk factor (overweight / obesity, high blood pressure, high blood sugar, high triglycerides levels, low HDL-C levels). The inventors demonstrated that Rab30 silencing improve metabolic dysfunction–associated steatotic liver disease (MASLD). The present invention relates to a method for the treatment of metabolic dysfunction–associated steatotic liver disease (MASLD) in a subject in need thereof comprising the administration of a therapeutically effective amount of an inhibitor of Rab30.

Description

[0001] INHIBITOR OF RAB30 FOR USE IN A METHOD OF TREATMENT OF METABOLIC DYSFUNCTION-ASSOCIATED STEATOTIC LIVER DISEASE (MASLD)

[0002] FIELD OF THE INVENTION:

[0003] The present invention is in the field of medicine, in particular hepatology.

[0004] BACKGROUND OF THE INVENTION:

[0005] Metabolic dysfunction-associated steatotic liver disease (MASLD), previously known as nonalcoholic fatty liver disease (NAFLD), is a condition characterized by the accumulation of fat in the liver of individuals with no alcohol consumption, and at least one cardiometabolic risk factor (overweight / obesity, high blood pressure, high blood sugar, high triglycerides levels, low HDL-C levels). MASLD encompasses a spectrum of liver conditions, ranging from simple fatty liver (steatosis) to a more severe form called metabolic dysfunction-associated steatohepatitis (MASH). In simple fatty liver, there is an accumulation of excess fat in the liver cells, which does not typically cause significant liver damage. However, in MASH, in addition to fat accumulation, inflammation and liver cell injury are present. Over time, MASH can progress to liver fibrosis (scarring), cirrhosis (severe scarring), and even liver failure. The exact cause of MASLD is not fully understood, but it is closely associated with certain risk factors, including obesity, insulin resistance (prediabetes or type 2 diabetes), high blood pressure, high cholesterol levels, and a sedentary lifestyle. Genetic factors and certain medications can also contribute to the development of MASLD. MASLD often does not cause noticeable symptoms in the early stages. However, as the disease progresses, individuals may experience fatigue, abdominal discomfort, and jaundice (yellowing of the skin and eyes) in severe cases. MASLD is typically detected through routine blood tests or imaging studies, such as ultrasound, computed tomography (CT), or magnetic resonance imaging (MRI). Management of MASLD involves lifestyle modifications aimed at reducing risk factors. This includes maintaining a healthy weight through diet and exercise, managing blood sugar levels, lowering cholesterol and blood pressure, and avoiding alcohol and unnecessary medications. In more advanced stages of the disease, additional interventions may be required to manage complications or treat liver fibrosis and cirrhosis. This is still a need to find new treatment of metabolic dysfunction-associated steatotic liver disease (MASLD). In this context the inventors demonstrated that Rab30 silencing improve metabolic dysfunction-associated steatotic liver disease (MASLD).

[0006] SUMMARY OF THE INVENTION:

[0007] The present invention is defined by the claims. In particular, the present invention relates to a method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof comprising the administration of a therapeutically effective amount of an inhibitor of Rab30.

[0008] DETAILED DESCRIPTION OF THE INVENTION:

[0009] Method of treatment:

[0010] In a first embodiment, the present invention relates to a method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof comprising the administration of a therapeutically effective amount of an inhibitor of Rab30.

[0011] As used herein, the term “subject” or “patient” refer to any mammals, such as a rodent, a feline, a canine, and a primate. In a particular embodiment, the subject is human. In a particular embodiment, the subject is a newborn or a child or teenager or an adult. In some embodiment, the subject of the present invention has a liver disease. Particularly, in the present invention, the subject has or is susceptible to have a metabolic dysfunction-associated steatotic liver disease (MASLD).

[0012] As used herein, the term “liver disease” refers to any disturbance of liver function that causes illness. The liver is responsible for many critical functions from protein production and blood clotting to cholesterol, glucose (sugar), and iron metabolism. When it becomes diseased or injured, the loss of those functions can cause significant damage to the body. Liver disease is also referred to as hepatic disease. The liver disease is selected from the group consisting of metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction- associated steatohepatitis (MASH), fibrotic MASH or liver cancer.

[0013] In the context of the invention, the liver disease is MASLD. As used herein, the term “metabolic dysfunction-associated steatotic liver disease” (MASLD), previously known as non-alcoholic fatty liver disease (NAFLD), is a condition characterized by the accumulation of fat in the liver of individuals with no alcohol consumption, and at least one cardiometabolic risk factor (overweight / obesity, high blood pressure, high blood sugar, high triglycerides levels, low HDL-C levels). The term metabolic dysfunction- associated steatotic liver disease (MASLD) encompasses the term non-alcoholic fatty liver disease (NAFLD). MASLD can be sub-classified as non-alcoholic steatohepatitis (MASH) and nonalcoholic fatty liver (NAFL).

[0014] In some embodiment, the MASLD is a nonalcoholic fatty liver (NAFL).

[0015] As used herein, the term “nonalcoholic fatty liver” (NAFL) is a type of MASLD and is a condition in which fat accumulates in the liver cells. NAFL has minimal risk of progressing to cirrhosis.

[0016] In some embodiment, the MASLD is a metabolic dysfunction-associated steatohepatitis (MASH).

[0017] As used herein, the term “metabolic dysfunction-associated steatohepatitis” (MASH) previously known as non-alcoholic steatohepatitis (NASH). In simple fatty liver, there is an accumulation of excess fat in the liver cells, which does not typically cause significant liver damage. However, in MASH, in addition to fat accumulation, inflammation and liver cell injury are present. The term metabolic dysfunction-associated steatohepatitis (MASH) encompasses the term non-alcoholic steatohepatitis (NASH).

[0018] In some embodiments, the patient has a cirrhosis.

[0019] As used herein, the term “cirrhosis” is used herein to refer to a pathologic liver condition characterized anatomically by widespread nodules in the liver combined with fibrosis. Cirrhosis represents the final common pathway for must types of chronic liver diseases, including those associated with chronic alcohol abuse, chronic viral hepatitis, metabolic and biliary diseases.

[0020] In some embodiments, the patient has a fibrosis. As used herein, “fibrosis” refers to the formation of excess fibrous connective tissue as a result of the excess deposition of extracellular matrix components, for example collagen. Fibrous connective tissue is characterised by having extracellular matrix (ECM) with a high collagen content. The collagen may be provided in strands or fibers, which may be arranged irregularly or aligned. The ECM of fibrous connective tissue may also include glycosaminoglycans.

[0021] In some embodiments, the subject suffers from obesity.

[0022] As used herein, the term "obesity" refers to a condition characterized by an excess of body fat having health consequences. The operational (or clinical) definition of obesity is based on the Body Mass Index (BMI), which is calculated as body weight per height in meter squared (kg / m2). Obesity refers to a condition whereby a subject has a BMI greater than or equal to 30 kg / m2, and overweight refers to a condition whereby a subject has a BMI greater than or equal to 25 kg / m2. The increased risks associated with obesity may occur at a lower BMI in people of Asian descent. In Asian and Asian-Pacific countries, including Japan, "obesity" refers to a condition whereby a subject has a BMI greater than or equal to 25 kg / m2. An "obese subject" in these countries refers to a subject with at least one obesity - induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, with a BMI greater than or equal to 25 kg / m2.

[0023] As used herein, the term "high fat diet" (HFD) is used herein to refer to diets that typically have a higher percentage of calories (obtained from the fat in the diet) compared to a normal diet. The fat in the diet may include all types of dietary fat, whether animal or vegetable and whether monounsaturated, polyunsaturated, saturated, etc. Thus, the high fat diet has a higher caloric content than a normal diet. In some embodiments, the non-human transgenic animal is fed with a diet having equal to or greater than 30% of total energy from fat. In some embodiments, the non-human transgenic animal is fed with a diet having equal to or greater than 35%, 40%, 45%, 50%, 55%, 60%, or 65% of total energy from fat. For instance, the high fat diet contains 414.0 kcal / 100 g with 43% as carbohydrate, 17% as protein, and 40% as fat. Another example of diet contains 23% of fat (e.g. butter), 17% as protein, and 0,2% of cholesterol.

[0024] As used herein, the terms “treating” or “treatment” refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of subject at risk of contracting the disease or suspected to have contracted the disease as well as subject who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By "therapeutic regimen" is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a subject during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a subject during treatment of an illness, e.g., to keep the subject in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., pain, disease manifestation, etc.]).

[0025] In some embodiment, the present invention relates to an inhibitor of Rab30 for use in the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof.

[0026] As used herein, the term “Rab30” is a member RAS oncogene family and refers to a protein that in humans is encoded by the RAB30 gene. Rab30 is predicted to enable GTP binding activity and GTPase activity and is involved in Golgi organization and endocytic trafficking function. Rab30 is located on endosomes, Golgi cisterna; cis-Golgi network; and trans-Golgi network. Rab30 is having the following human Gene ID: 27314 and the following human UniProt number QI 5771. As used herein, the term “inhibitor” or “antagonist” refers to a compound that decreases the magnitude of at least one activity, signaling or expression of a molecule compared to the magnitude of the activity, signaling or expression observed in the absence of the inhibitor. In some instances, an inhibitor will substantially decrease the magnitude of at least one activity, signaling or expression of a molecule compared to the magnitude of the activity or expression observed in the absence of the inhibitor. In some instances, an inhibitor will completely diminish the magnitude of at least one activity, signaling or expression of a molecule compared to the magnitude of the activity, signaling or expression observed in the absence of the inhibitor. Certain exemplary inhibitors include, but are not limited to, proteins, peptides, antibodies, peptibodies, aptamers, antisense oligonucleotides, interfering RNA, carbohydrates or small organic molecules.

[0027] As used herein, an "inhibitor of expression" refers to a natural or synthetic compound that has a biological effect to inhibit the expression of a gene. In a particular embodiment of the invention, the inhibitor of gene expression is a siRNA, an antisense oligonucleotide or a ribozyme. For example, anti- sense oligonucleotides, including anti-sense RNA molecules and anti-sense DNA molecules, would act to directly block the translation of Rab30 mRNA by binding thereto and thus preventing protein translation or increasing mRNA degradation, thus decreasing the level of Rab30, and thus activity, in a cell. For example, antisense oligonucleotides of at least about 15 bases and complementary to unique regions of the mRNA transcript sequence encoding Rab30 can be synthesized, e.g., by conventional phosphodiester techniques. Methods for using antisense techniques for specifically inhibiting gene expression of genes whose sequence is known are well known in the art (e.g. see U.S. Pat. Nos. 6,566, 135; 6,566,131; 6,365,354; 6,410,323; 6,107,091; 6,046,321; and 5,981,732). Small inhibitory RNAs (siRNAs) can also function as inhibitors of expression for use in the present invention. Rab30 gene expression can be reduced by contacting a subject or cell with a small double stranded R A (dsPvNA), or a vector or construct causing the production of a small double stranded R A, such that Rab30 gene expression is specifically inhibited (i.e. RNA interference or RNAi). Antisense oligonucleotides, siRNAs, shRNAs and ribozymes of the invention may be delivered in vivo alone or in association with a vector. In its broadest sense, a "vector" is any vehicle capable of facilitating the transfer of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid to the cells and typically cells expressing Rab30. Typically, the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector. In general, the vectors useful in the invention include, but are not limited to, plasmids, phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid sequences. Viral vectors are a preferred type of vector and include, but are not limited to nucleic acid sequences from the following viruses: retrovirus, such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma virus; adenovirus, adeno-associated virus; SV40-type viruses; polyoma viruses; Epstein-Barr viruses; papilloma viruses; herpes virus; vaccinia virus; polio virus; and RNA virus such as a retrovirus. One can readily employ other vectors not named but known to the art. In some embodiments, the inhibitor consists in a vector that comprises the CRISPR / cas 9 protein and the appropriate RNA guide for disrupting the expression level of the gene encoding for Rab30.

[0028] In a particular embodiment, the inhibitor of Rab30 is a peptide, peptidomimetic, small organic molecule, antibody, aptamers, siRNA or antisense oligonucleotide. The term “peptidomimetic” refers to a small protein-like chain designed to mimic a peptide.

[0029] In a particular embodiment, the inhibitor of Rab30 is an aptamer. Aptamers are a class of molecule that represents an alternative to antibodies in term of molecular recognition. Aptamers are oligonucleotide or oligopeptide sequences with the capacity to recognize virtually any class of target molecules with high affinity and specificity.

[0030] In some embodiments, the inhibitor of Rab30 is a short hairpin RNA (shRNA), a small interfering RNA (siRNA) or an antisense oligonucleotide which inhibits the expression of metabolites involved in Rab30 metabolism.

[0031] In a particular embodiment, the inhibitor of Rab30 is a siRNA. A short hairpin RNA (shRNA) is a sequence of RNA that makes a tight hairpin turn that can be used to silence gene expression via RNA interference. shRNA is generally expressed using a vector introduced into cells, wherein the vector utilizes the U6 promoter to ensure that the shRNA is always expressed. This vector is usually passed on to daughter cells, allowing the gene silencing to be inherited. The shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC). This complex binds to and cleaves mRNAs that match the siRNA to which it is bound. Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, are a class of 20-25 nucleotide-long double- stranded RNA molecules that play a variety of roles in biology. Most notably, siRNA is involved in the RNA interference (RNAi) pathway whereby the siRNA interferes with the expression of a specific gene.

[0032] In a particular embodiment, the inhibitor of Rab30 is an anti-sense oligonucleotides (ASO). Anti-sense oligonucleotides include anti-sense RNA molecules and anti-sense DNA molecules, would act to directly block the translation of the targeted mRNA by binding thereto and thus preventing protein translation or increasing mRNA degradation, thus decreasing the level of the targeted protein, and thus activity, in a cell. For example, antisense oligonucleotides of at least about 15 bases and complementary to unique regions of the mRNA transcript sequence can be synthesized, e.g., by conventional phosphodiester techniques. Methods for using antisense techniques for specifically inhibiting gene expression of genes whose sequence is known are well known in the art (e.g. see U.S. Pat. Nos. 6,566,135; 6,566,131; 6,365,354; 6,410,323; 6,107,091; 6,046,321; and 5,981,732). Antisense oligonucleotides, siRNAs, shRNAs of the invention may be delivered in vivo alone or in association with a vector. In its broadest sense, a "vector" is any vehicle capable of facilitating the transfer of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid to the cells. Typically, the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector. In general, the vectors useful in the invention include, but are not limited to, plasmids, phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid sequences. Viral vectors are a preferred type of vector and include, but are not limited to nucleic acid sequences from the following viruses: retrovirus, such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma virus; adenovirus, adeno-associated virus; SV40-type viruses; polyoma viruses; Epstein-Barr viruses; papilloma viruses; herpes virus; vaccinia virus; polio virus; and RNA virus such as a retrovirus. One can readily employ other vectors not named but known to the art.

[0033] In some embodiments, the inhibitor of Rab30 is an endonuclease. In the last few years, staggering advances in sequencing technologies have provided an unprecedentedly detailed overview of the multiple genetic aberrations in cancer. By considerably expanding the list of new potential oncogenes and tumor suppressor genes, these new data strongly emphasize the need of fast and reliable strategies to characterize the normal and pathological function of these genes and assess their role, in particular as driving factors during oncogenesis. As an alternative to more conventional approaches, such as cDNA overexpression or downregulation by RNA interference, the new technologies provide the means to recreate the actual mutations observed in cancer through direct manipulation of the genome. Indeed, natural and engineered nuclease enzymes have attracted considerable attention in the recent years. The mechanism behind endonuclease-based genome inactivating generally requires a first step of DNA single or double strand break, which can then trigger two distinct cellular mechanisms for DNA repair, which can be exploited for DNA inactivating: the errorprone nonhomologous end-joining (NHEJ) and the high-fidelity homology-directed repair (HDR).

[0034] In a particular embodiment, the endonuclease is CRISPR-cas. As used herein, the term “CRISPR-cas” has its general meaning in the art and refers to clustered regularly interspaced short palindromic repeats associated which are the segments of prokaryotic DNA containing short repetitions of base sequences.

[0035] In some embodiment, the endonuclease is CRISPR-cas9 which is from Streptococcus pyogenes. The CRISPR / Cas9 system has been described in US 8697359 Bl and US 2014 / 0068797. Originally an adaptive immune system in prokaryotes (Barrangou and Marraffini, 2014), CRISPR has been recently engineered into a new powerful tool for genome editing. It has already been successfully used to target important genes in many cell lines and organisms, including human (Mali et al., 2013, Science, Vol. 339 : 823-826), bacteria (Fabre et al., 2014, PLoS Negl. Trop. Dis., Vol. 8:e2671.), zebrafish (Hwang et al., 2013, PLoS One, Vol. 8:e68708.), C. elegans (Hai et al., 2014 Cell Res. doi: 10.1038 / cr.2014.11.), bacteria (Fabre et al., 2014, PLoS Negl. Trop. Dis., Vol. 8:e2671.), plants (Mali et al., 2013, Science, Vol. 339 : 823-826), Xenopus tropicalis (Guo et al., 2014, Development, Vol. 141 : 707-714.), yeast (DiCarlo et al., 2013, Nucleic Acids Res., Vol. 41 : 4336-4343.), Drosophila (Gratz et al., 2014 Genetics, doi: 10.1534 / genetics.113.160713), monkeys (Niu et al., 2014, Cell, Vol. 156 : 836- 843.), rabbits (Yang et al., 2014, J. Mol. Cell Biol., Vol. 6 : 97-99.), pigs (Hai et al., 2014, Cell Res. doi: 10.1038 / cr.2014.11.), rats (Ma et al., 2014, Cell Res., Vol. 24 : 122-125.) and mice (Mashiko et al., 2014, Dev. Growth Differ. Vol. 56 : 122-129.). Several groups have now taken advantage of this method to introduce single point mutations (deletions or insertions) in a particular target gene, via a single gRNA. Using a pair of gRNA-directed Cas9 nucleases instead, it is also possible to induce large deletions or genomic rearrangements, such as inversions or translocations. A recent exciting development is the use of the dCas9 version of the CRISPR / Cas9 system to target protein domains for transcriptional regulation, epigenetic modification, and microscopic visualization of specific genome loci.

[0036] In some embodiment, the endonuclease is CRISPR-Cpfl which is the more recently characterized CRISPR from Provotella and Francisella 1 (Cpfl) in Zetsche et al. (“Cpfl is a Single RNA-guided Endonuclease of a Class 2 CRISPR-Cas System (2015); Cell; 163, 1-13).

[0037] In some embodiments, the inhibitor of Rab30 is an antibody. As used herein, the term “antibody” is used in the broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g. bispecific antibodies) formed from at least two intact antibodies, and antibody fragments so long as they exhibit the desired biological activity. The term includes antibody fragments that comprise an antigen binding domain such as Fab', Fab, F(ab')2, single domain antibodies (DABs), TandAbs dimer, Fv, scFv (single chain Fv), dsFv, ds-scFv, Fd, linear antibodies, minibodies, diabodies, bispecific antibody fragments, bibody, tribody (scFv-Fab fusions, bispecific or trispecific, respectively); sc-diabody; kappa(lamda) bodies (scFv-CL fusions); BiTE (Bispecific T-cell Engager, scFv-scFv tandems to attract T cells); DVD-Ig (dual variable domain antibody, bispecific format); SIP (small immunoprotein, a kind of minibody); SMIP ("small modular immunopharmaceutical" scFv-Fc dimer; DART (ds-stabilized diabody "Dual Affinity ReTargeting"); small antibody mimetics comprising one or more CDRs and the like. The techniques for preparing and using various antibody -based constructs and fragments are well known in the art (see Kabat et al., 1991, specifically incorporated herein by reference). Diabodies, in particular, are further described in EP 404, 097 and WO 93 / 1 1 161; whereas linear antibodies are further described in Zapata et al. (1995). Antibodies can be fragmented using conventional techniques. For example, F(ab')2 fragments can be generated by treating the antibody with pepsin. The resulting F(ab')2 fragment can be treated to reduce disulfide bridges to produce Fab' fragments. Papain digestion can lead to the formation of Fab fragments. Fab, Fab' and F(ab')2, scFv, Fv, dsFv, Fd, dAbs, TandAbs, ds-scFv, dimers, minibodies, diabodies, bispecific antibody fragments and other fragments can also be synthesized by recombinant techniques or can be chemically synthesized. Techniques for producing antibody fragments are well known and described in the art. For example, each of Beckman et al., 2006; Holliger & Hudson, 2005; Le Gall et al., 2004; Reff & Heard, 2001 ; Reiter et al., 1996; and Young et al., 1995 further describe and enable the production of effective antibody fragments. In some embodiments, the antibody is a “chimeric” antibody as described in U.S. Pat. No. 4,816,567. In some embodiments, the antibody is a humanized antibody, such as described U.S. Pat. Nos. 6,982,321 and 7,087,409. In some embodiments, the antibody is a human antibody. A “human antibody” such as described in US 6,075,181 and 6,150,584. In some embodiments, the antibody is a single domain antibody such as described in EP 0 368 684, WO 06 / 030220 and WO 06 / 003388.

[0038] In a particular embodiment, the inhibitor of Rab30 is a monoclonal antibody. Monoclonal antibodies can be prepared and isolated using any technique that provides for the production of antibody molecules by continuous cell lines in culture. Techniques for production and isolation include but are not limited to the hybridoma technique, the human B-cell hybridoma technique and the EBV-hybridoma technique.

[0039] In a particular embodiment, the inhibitor of Rab30 is an intrabody. As used herein, the term "intrabody" generally refer to an intracellular antibody or antibody fragment. Antibodies, in particular single chain variable antibody fragments (scFv), can be modified for intracellular localization. Such modification may entail for example, the fusion to a stable intracellular protein, such as, e.g., maltose binding protein, or the addition of intracellular trafficking / localization peptide sequences, such as, e.g., the endoplasmic reticulum retention. In some embodiments, the intrabody is a single domain antibody. In some embodiments, the antibody according to the invention is a single domain antibody. The term “single domain antibody” (sdAb) or "VHH" refers to the single heavy chain variable domain of antibodies of the type that can be found in Camelid mammals which are naturally devoid of light chains. Such VHH are also called “nanobody®”. According to the invention, sdAb can particularly be llama sdAb.

[0040] In a particular embodiment, the inhibitor of Rab30 is a small organic molecule. The term “small organic molecule” refers to a molecule of a size comparable to those organic molecules generally used in pharmaceuticals. The term excludes biological macromolecules (e.g., proteins, nucleic acids, etc.). Preferred small organic molecules range in size up to about 5000 Da, more preferably up to 2000 Da, and most preferably up to about 1000 Da.

[0041] As used herein the terms "administering" or "administration" refer to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g. an inhibitor of Rab30) into the subject, such as by oral, mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery and / or any other method of physical delivery described herein or known in the art. When a disease, or a symptom thereof, is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease or symptoms thereof, are being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof. In another embodiment, an oral administration is performed to the subject. In a further embodiment, intravenous administration is performed to the subject.

[0042] By a "therapeutically effective amount" is meant a sufficient amount of an inhibitor of Rab30 for use in a method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) at a reasonable benefit / risk ratio applicable to any medical treatment. It will be understood that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts. For example, it is well known within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day. Typically, the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic 20 adjustment of the dosage to the subject to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, typically from 1 mg to about 100 mg of the active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg / kg to about 20 mg / kg of body weight per day, especially from about 0.001 mg / kg to 7 mg / kg of body weight per day.

[0043] Typically the active ingredient of the present invention is combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form pharmaceutical compositions. The term "Pharmaceutically" or "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0044] Combined preparation:

[0045] In some embodiment the inhibitor of Rab30 is combined with a PPARa agonist.

[0046] As used herein, the term “PPARa” has its general meaning of the art and refers to the gene encoding for peroxisome proliferator-activated receptor alpha.

[0047] As used herein, the term “PPARa agonist” refers to drugs which act upon the peroxisome proliferator-activated receptor. They are used for the treatment of symptoms of the metabolic syndrome, mainly for lowering triglycerides and blood sugar.

[0048] PPARa agonists are drugs which act upon the peroxisome proliferator-activated receptor. They are used for the treatment of symptoms of the metabolic syndrome, mainly for lowering triglycerides and blood sugar. Example of PPARa agonists include but are not limited to Pemafibrate, Elafibranor, Muraglitazar, Lobeglitazone, Naveglitazar.

[0049] As used herein, the terms “combined treatment”, “combined therapy” or “therapy combination” refer to a treatment that uses more than one medication. The combined therapy may be dual therapy or bi-therapy.

[0050] As used herein, the term “administration simultaneously” refers to administration of 2 active ingredients by the same route and at the same time or at substantially the same time. The term “administration separately” refers to an administration of 2 active ingredients at the same time or at substantially the same time by different routes. The term “administration sequentially” refers to an administration of 2 active ingredients at different times, the administration route being identical or different.

[0051] Pharmaceutical composition: Accordingly, in a further aspect, the invention relates to a pharmaceutical composition comprising an inhibitor of Rab30 for use in a method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof.

[0052] In a particular embodiment, the pharmaceutical composition according the invention, wherein the inhibitor of Rab30 is a peptide, peptidomimetic, small organic molecule, antibody, aptamers, siRNA or antisense oligonucleotide.

[0053] As used herein, the terms "pharmaceutically" or "pharmaceutically acceptable" refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. The pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, the active principle, alone or in combination with another active principle, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms. Typically, the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The polypeptide (or nucleic acid encoding thereof) can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin. Sterile injectable solutions are prepared by incorporating the active polypeptides in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuumdrying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermiclysis fluid or injected at the proposed site of infusion. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

[0054] Method for screening:

[0055] In a further aspect, the invention relates to a method of screening a drug suitable for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) comprising i) providing a test compound and ii) determining the ability of said test compound to inhibit the expression or activity of Rab30.

[0056] Any biological assay well known in the art could be suitable for determining the ability of the test compound to inhibit the activity or expression of Rab30. In some embodiments, the assay first comprises determining the ability of the test compound to bind to Rab30. In some embodiments, a population of cells then contacted and activated so as to determine the ability of the test compound to inhibit the activity or expression of Rab30. In particular, the effect triggered by the test compound is determined relative to that of a population of immune cells incubated in parallel in the absence of the test compound or in the presence of a control agent either of which is analogous to a negative control condition. The term "control substance", "control agent", or "control compound" as used herein refers a molecule that is inert or has no activity relating to an ability to inhibit a biological activity or expression. It is to be understood that test compounds capable of inhibiting the activity or expression of Rab30, as determined using in vitro methods described herein, are likely to exhibit similar modulatory capacity in applications in vivo. Typically, the test compound is selected from the group consisting of peptides, peptidomimetics, small organic molecules, antibodies (e.g. intraantibodies), aptamers or nucleic acids. For example the test compound according to the invention may be selected from a library of compounds previously synthesised, or a library of compounds for which the structure is determined in a database, or from a library of compounds that have been synthesised de novo. The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.

[0057] FIGURES:

[0058] Figure 1: A) Rab30 mRNA level in liver from 5h fasted (black bar) or random fed (white bar) mice. ** pvalue < 0.01. B) Rab30 mRNA level in liver from wild type mice (PPARaWT - black bars) or PPARa hepatocyte specific knockout mice (PPARaHEPKO - white bars), treated with the vehicle or pemafibrate. *** pvalue < 0.001. C) Rab30 mRNA level in primary mouse hepatocytes treated with the vehicle (black bar), pemafibrate (grey bar) or a mixture of Palmitate and Oleate (PA:OA - white bar). * pvalue < 0.05 , *** pvalue < 0.001. D) Rab30 mRNA level in the liver from control (Chow diet - black bar) or STAM mice (a common mouse model of MASLD - white bar). *** pvalue < 0.001. E) Rab30 mRNA level in the liver from control (Chow diet - black bar) or mice fed with a Western Diet High Fructose (a common mouse model of MASLD - white bar). * pvalue < 0.05. F) Representative image of a Westemblot showing Rab30 (top panel) and ERK2 (bottom panel) protein levels in primary mouse hepatocytes transfected with Interferin complexed either with a siRNA directed against a Scrambled sequence (siScrble - control) or Rab30 (siRab30). G) Representative light microscopy image of Bodipy stained primary mouse hepatocyte expressing (siScrble) or not Rab30 (siRab30) following a 24h-exposure to a mixture of palmitate and oleate fatty acids in presence of the PPARa agonist pemafibrate. H) Triglyceride levels in primary mouse hepatocytes expressing (siScrble - black bars) or not Rab30 (siRab30 - white bars) and exposed to BSA (vehicle) or a mixture of Palmitate and Oleate (PA / OA) in presence of the PPARa agonist pemafibrate for 24h. *** pvalue < 0.001. I) Time course of TF2-C12 accumulation in primary mouse hepatocytes expressing (siScrble - black bars) or not Rab30 (siRab30 - white bars) pretreated with pemafibrate for 24h. J) Heatmap of mRNA expression of selected genes involved in P-oxydation in primary mouse hepatocytes expressing (siScrble - black bars) or not Rab30 (siRab30 - white bars) and exposed to BSA (vehicle) or a mixture of Palmitate and Oleate (PA / OA) in presence of the PPARa agonist pemafibrate for 24h.

[0059] Figure 2: A) Rab30 mRNA level in liver from normal chow diet (ND) random-fed C57 / B16 mice three days after intravenous injection of lipid nanoparticles loaded with siRNA directed against a scramble sequence (siScrble - control - black bar) or Rab30 (siRab30 - white bar). * pvalue < 0.05. B) Liver and subcutaneous adipose tissue (scWAT) weight at sacrifice from ND- fed control mice (siScrble - black bar) or mice silenced for Rab30 (siRab30 - white bar) three days after intravenous injection of lipid particles loaded with the siRNAs. ** pvalue < 0.01. C) Triglycerides in liver at sacrifice from ND-fed control mice (siScrble - black bar) or mice silenced for Rab30 (siRab30 - white bar) three days after intravenous injection of lipid particles loaded with the siRNAs. * pvalue < 0.05. D) Rab30 mRNA level in liver from C57 / B16 mice fed 12 weeks with a Western-diet (WD) four days after intravenous injection of lipid nanoparticles loaded with siRNA directed against a scramble sequence (siScrble - control - black bar) or Rab30 (siRab30 - white bar). * pvalue < 0.05. E) Blood insulin level in ND- or WD-fed mice treated as described in (A) and (D) respectively. Diet effect : $ pvalue < 0.05. Silencing effect : * pvalue < 0.05. ns = statistically non-significant. ND = mice fed a normal chow diet ; WD = mice fed for 12 weeks with a western diet.

[0060] EXAMPLE:

[0061] Material & Methods

[0062] Mice

[0063] Fasting / feeding experiments: C57B16 mice were housed in a 23°C on a 12 h light-dark cycle in the animal facility. 15-weeks male mice were sacrificed either random fed (at ZT2) or after a 5h fasting (at ZT7). Livers were harvested and a RNAseq analysis was performed at BGI Genomics.

[0064] In vivo pemafibrate experiments: Ten-week old male and female Pparahep+ / + and Pparahep- / - mice (n=8 per group) fed a standard diet (Safe A04) received 0.1 mg / kg / day pemafibrate (Kowa, MedChemExpress) or vehicle (carboxymethylcellulose 0.5%) during 14 days. Gavages were performed every day at ZT16. Mice were sacrificed at ZT16, fed ad libitum, the last gavage was performed the day before the sacrifice.

[0065] MASLD mouse models:

[0066] StamMice: Mice were obtained from the SMC Laboratories. Briefly, C57BL / 6J mice (14-day- pregnant female, 6 weeks of age male) have been obtained from Japan SLC, Inc. (Japan). The animals have been maintained in a SPF facility under controlled conditions of temperature (23 ± 3°C}, humidity (50± 20%), lighting (12-hour artificial light and dark cycles; light from 8:00 to 20:00) and air exchange. MASH was induced in male mice by a single subcutaneous injection of 200 g streptozotocin (STZ, Sigma- Aldrich, USA) solution 2 days after birth and feeding with high fat diet (HFD, 57 kcal% fat, Cat# HFD32, CLEA Japan, Inc., Japan) after 4 weeks of age. The animals were sacrificed at 24 weeks of age by exsanguination through direct cardiac puncture under isoflurane anesthesia (Pfizer Inc.).

[0067] WD-High fructose: Ten-week-old wild-type male mice (Janvier Labs, Le Genest-Saint-Isle, France) were acclimated to our animal facilities under a 12-hour / 12-hour light / dark cycle at a temperature of 21 °C ± 2°C and were fed ad libitum either a Western Diet (diet from Ssniff #S8926-E060 EF Western diet, +0.5 % Cholesterol, custom-made) or chow diet (CD) and the drinking water was supplemented with 15% fructose for 27 weeks.

[0068] Liver Rab30 silencing:

[0069] After 12 weeks of normal chow or Western diet, mice were randomized into 4 groups (ND siScrble, ND siRab30, WD siScrble, WD siRab30) and intravenously injected with lipid nanoparticles (InVivoFectamin 3.0, ThermoFisher) pre-loaded with siRNAs directed against Rab30 or a scramble sequence (Ambion in vivo siRNA, ThermoFisher), following company's recommendations. Blood was collected at sacrifice. Tissues were harvested at sacrifice, weighted and snap frozen in liquid nitrogen.

[0070] Metabolic exploration:

[0071] For liver triglycerides: small livers pieces were weighted, lyzed in 5% NP40 and heated at 100°C. After centrifugation, supernatants were collected, and triglyceride levels were estimated using the Diasys Triglyceride assay kit.

[0072] For insulinemia: serum insulin levels were measured using an HTRF-based assay (Revity).

[0073] Primary hepatocyte culture and silencing

[0074] Primary murine hepatocytes preparation: Male and female C57B16 wild type mice were anesthetized and perfused via the vena cava with the washing buffer (HBSS, lOmM Hepes and 0.5mM EGTA) and the dissociation buffer (HBSS, lOmM Hepes, 0.4mM CaC12 and 120 U / mL Collagenase). Livers were collected, dissociated in seeding medium (William’s media, 10% FCS, 2mM L-Glutamin, 1% penicillin / streptomycin, and 130nM insulin) and filtered through 100pm filters. Cells were centrifuged at 50g on a 90% Percoll gradient to collect live hepatocytes in the pellet. Primary mouse hepatocytes in the pellets were resuspended in seeding medium at 400,000 cells / ml and cultured at 37°C and 5% CO2 in a humid chamber. Rab30 silencing: Primary mouse hepatocytes were incubated for 6h with OptiMEM media containing Interferin and siRNA (directed against a scrambled sequence or Rab30), following customer recommendations.

[0075] Hepatocytes treatments: Fasting mimicking media (DMEM, ImM de glucose, ImM pyruvate, 2.5mM glycerol, 2mM L-Glutamin, lOpM Forskolin, Img / ml BSA and 1% penicillin / streptomycin) was complemented with lOpM pemafibrate and / or a mixture of fatty acids (0.33mM Palmitate + 0.66mM Oleate). Primary mouse hepatocytes were exposed to these different treatments for 24h.

[0076] Fluorescence-based assays

[0077] BodiPy staining and imaging: Following the 24h exposure of the primary mouse hepatocytes to Palmitate / Oleate and pemafibrate, cells were fixed with 4% PFA for 30min at room temperature. Lipid droplets were stained with 2pM BodiPy for 20min at room temperature. Unbiased automated images were acquired on a Nikon confocal AIR microscope equipped with a 40x water immersion objective.

[0078] Fluorescent TF2-C12 uptake assay: Primary mouse hepatocytes treated for 24h with pemafibrate were exposed to the TF2-coupled dodecanoic acid (TF2-C12). Real time fluorescence was measured for 60 min on a fluorometer (kex=485nm & kem=515 nm) at 37°C.

[0079] RNAseq analyses

[0080] Mice livers and primary mouse hepatocytes were snap frozen in LN2. Samples were sent to BGI Genomics, which performed the RNA extraction, the library generation, the sequencing, the normalization, and the analysis.

[0081] Statistical analyses

[0082] All the data were analyzed using GraphPad Prism7 software. For multiple comparisons, ANOVA tests were performed, followed by a Bonferroni post-hoc test. When comparing two independent groups, a Shapiro-wilk test was systematically performed to determine whether the distribution was symmetrical or asymmetrical. For asymmetrical distributions, a Mann & Whitney test was performed. For symmetrical distributions, an F-test was performed to determine whether the variances are homogeneous or heterogeneous; if the variances were homogeneous, a Student's T-test was performed; if they were heterogeneous, Welch's correction to the Student's test was performed. In all cases, variations were considered significant if the Pvalue was lower than 0.05.

[0083] Results

[0084] Our results demonstrates that the expression of Rab30 in mouse liver is induced during fasting (Figure 1A). This induction of Rab30 expression in vivo is under the control of the transcription factor PPARa (Figure IB), which is the master regulator of liver response to fasting. In primary mouse hepatocyte, a cell autonomous system, the expression of Rab30 is induced in response to the PPARa agonist pemafibrate and to a mixture of palmitate and oleate fatty acids (Figure 1C). Overall, these results established that Rab30 expression is controlled by PPARa in response to fatty acids. The expression of Rab30 mRNA was also found induced by 4- and 2- folds in the liver of two mouse models of Metabolic dysfunction-Associated Steatotic Liver Disease (MASLD): the STAM mice (Figure ID), and the Western Diet high fructose mice (Figure IE). To determine the role of Rab30, we first depleted Rab30 in primary mouse hepatocyte by an RNAi-based approach (Figure IF). Next, the control (siScrble) or Rab30 depleted (siRab30) primary mouse hepatocytes were exposed to a mixture of palmitate and oleate fatty acids to induce hepatocyte steatosis (Figures 1G-1H). The quantity of lipid droplets detected by Bodipy staining (Figure 1G) as well as the level of triglycerides in the cells (Figure 1H) were reduced in Rab30-depleted hepatocytes when compared to control hepatocytes. This reduction of Triglyceride is not due to a defect in fatty acids uptake, since the capacity of primary mouse hepatocytes expressing (siScrble - black line) or not Rab30 (siRab30 - grey line) to internalize fluorescently-labelled fatty acid (TF2-C12) are equivalent (Figure II). In fact, we found that the silencing of Rab30 in pemafibrate-treated primary mouse hepatocytes, to activate PPARa, favor the expression of numerous genes involved in fatty acid betaoxidation (Figure 1J), which could explain the reduction in triglyceride accumulation.

[0085] To determine the impact of Rab30 silencing in vivo, we injected intravenously lipid nanoparticles loaded with siScrble (used as control) or siRab30, in C57 / B16 mice fed a normal chow diet. Three days after injection, the level of Rab30 mRNA is significantly reduced in the liver of siRab30-treated mice (Figure 2A). These mice present a similar liver weight, but a significant reduction in subcutaneous adipose tissue (Figure 2B). Interestingly, the triglyceride amount in the liver is significantly reduced after three days of Rab30 silencing (Figure 2C). In accordance with our in vitro data, our in vivo results demonstrate that the silencing of Rab30 in the liver in vivo also reduce the triglycerides levels. To determine whether Rab30 inhibition could improve Metabolic Dysfunction- Associated Steatotic Liver Disease (MASLD), Rab30 was silenced in mice fed a Western diet for 12 weeks (Figure 2D), following the same procedure as for mice fed a normal diet. As expected, 12 weeks of Western diet increase random-fed insulin levels (Figure 2E), suggesting insulin resistance. Four days of Rab30 silencing in the liver of mice fed a Western diet for 12 weeks is sufficient to restore a level of insulin similar to those of normal chow diet mice (Figure 2E).

[0086] REFERENCES:

[0087] Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

Claims

- 23 -CLAIMS:

1. A method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof comprising the administration of a therapeutically effective amount of an inhibitor of Rab30.

2. The method of claim 1 wherein the inhibitor of Rab30 is a siRNA.

3. The method of claim 1 wherein the inhibitor of Rab30 is an antisense oligonucleotide.

4. The method of claim 1 wherein the inhibitor of Rab30 is a small molecule.

5. The method of claim 1 wherein the inhibitor of Rab30 is an antibody.

6. The method of claim 1 wherein the subject suffers from obesity.

7. The method of claim 1 wherein the MASLD is a non-alcoholic steatohepatitis (MASH).

8. The method of claim 1 wherein the MASLD is a nonalcoholic fatty liver (NAFL).

9. The method of claim 1 wherein the inhibitor of Rab30 is combined with a PPARa agonist.

10. The method of claim 9 wherein the PPARa agonist is Pemafibrate, Elafibranor, Muraglitazar, Lobeglitazone or Naveglitazar.

11. A pharmaceutical composition comprising an inhibitor of Rab30 for use in a method for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD) in a subject in need thereof.

12. The pharmaceutical composition according to claim 11 wherein the inhibitor of Rab30 is a peptide, peptidomimetic, small organic molecule, antibody, aptamers, siRNA or antisense oligonucleotide.

13. A method of screening a drug suitable for the treatment of MASLD comprising i) providing a test compound and ii) determining the ability of said test compound to inhibit the activity of Rab30.

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