Invented by Masanori Aikawa, Daiju Fukuda, Tetsuro Miyazaki, Elena Aikawa, Brigham and Womens Hospital Inc

Nonalcoholic fatty liver disease (NAFLD) is a growing health concern worldwide. It is a condition where excess fat accumulates in the liver, leading to inflammation and damage. NAFLD is often associated with obesity, insulin resistance, and metabolic syndrome. It can progress to nonalcoholic steatohepatitis (NASH), a more severe form of the disease that can lead to liver fibrosis, cirrhosis, and even liver cancer. Currently, there are no approved drugs for the treatment of NAFLD or NASH, and lifestyle changes such as weight loss and exercise remain the mainstay of therapy. However, recent research has identified a promising target for drug development in NAFLD and NASH: the Notch signaling pathway. The Notch signaling pathway is a highly conserved pathway that plays a critical role in cell fate determination, differentiation, and proliferation. It is involved in various physiological processes, including embryonic development, tissue homeostasis, and immune response. Dysregulation of the Notch pathway has been implicated in the pathogenesis of several diseases, including cancer, cardiovascular disease, and liver disease. In NAFLD and NASH, activation of the Notch pathway has been shown to contribute to liver inflammation, fibrosis, and steatosis. Inhibition of the Notch pathway has been proposed as a potential therapeutic strategy for NAFLD and NASH. Several preclinical studies have demonstrated the efficacy of Notch inhibition in the treatment and prevention of NAFLD and NASH. In animal models of NAFLD and NASH, pharmacological inhibition of the Notch pathway has been shown to reduce liver inflammation, fibrosis, and steatosis. Notch inhibition has also been shown to improve glucose and lipid metabolism, suggesting a potential role in the treatment of metabolic syndrome. Furthermore, genetic studies in humans have identified genetic variants in the Notch pathway that are associated with NAFLD and NASH, providing further evidence for the involvement of the Notch pathway in the pathogenesis of these diseases. Several pharmaceutical companies are currently developing Notch inhibitors for the treatment of NAFLD and NASH. These inhibitors include small molecules, antibodies, and RNA-based therapeutics. Some of these inhibitors have shown promising results in preclinical studies and are currently in clinical trials. For example, a phase 2 clinical trial of a Notch inhibitor in patients with NASH is currently underway, with preliminary results showing a reduction in liver fat content and improvement in liver enzymes. In conclusion, the Notch signaling pathway represents a promising target for the development of drugs for the treatment and prevention of NAFLD and NASH. Preclinical studies have demonstrated the efficacy of Notch inhibition in reducing liver inflammation, fibrosis, and steatosis, as well as improving glucose and lipid metabolism. Several pharmaceutical companies are currently developing Notch inhibitors for the treatment of NAFLD and NASH, and early clinical trials have shown promising results. Further research is needed to fully understand the role of the Notch pathway in NAFLD and NASH and to develop safe and effective drugs for these diseases.

The Brigham and Womens Hospital Inc invention works as follows

The present invention relates to nonalcoholic fatty liver disease treatment and prevention by using agents that inhibit NOTCH signaling pathways. This purpose may be achieved by using antibodies that inhibit the binding Delta like 4 ligand (Dll4) onto NOTCH receptors.

Background for Notch inhibition in nonalcoholic fatty liver diseases treatment and prevention

Obesity is a common condition that can be linked to other metabolic disorders and poses a threat to the health of people around the world. Over two-thirds (33%) of Americans are obese and over half (32%) are overweight. Atherosclerosis and heart attack are the most severe and common complications of obesity.

Metabolic Syndrome (Sutherland, Metabolic Syndrome & Related Disorders 2 to82-104 (2004); Esposito, Nutr. Metab. Cardiovasc. Dis. 14:228-228-232 (2004)). It is related to obesity and includes the following metabolic risk factors: 1) abdominal obesity (excessive body fat); 2) atherogenic dyslipidemia; high triglycerides, low HDL cholesterol, and high LDL cholesterol; 3) insulin resistance or glucose intolerance; and 5) prothrombotic (e.g. high fibrinogen or plasmaminogen activator inhibitor-1 levels in the blood); and 6 ) a proinflammatory (e.g. In developed countries, metabolic syndrome is becoming more common and closely linked with the risk of coronary disease (Malik, Irabarren, and others, Circulation 110, 1245-1250 (2004)). Am. Coll. Cardiol. 48:1800-1807 (2006)). There is mounting evidence that metabolic syndrome features are closely linked to chronic inflammation. These include macrophage accumulation, increased production of cytokine, and activation a network inflammatory signaling pathways.

Cardiometabolic Syndrome” includes obesity-related metabolic diseases and atherosclerosis. Cardiometabolic diseases can also cause arterial and valvular calciumification, which can lead to severe clinical complications such as acute myocardial injury and aortic narrowing. Diabetes can also cause chronic kidney disease, which can lead to cardiovascular ectopic calciumification and acute myocardial injury. The cardiometabolic syndrome is made up of several components that are interrelated and work together through systemic or local inflammation.

The NOTCH signaling pathway was identified as an important part of many biological functions including cell differentiation and proliferation (see U.S. Pat. No. 6,703,221). There have been many cases of leukemia-related mutations in genes that increase NOTCH signaling. Inhibitors of NOTCH are currently being investigated for their possible use in treating neurological diseases and cancer. J. Cancer 92:751-759 (2005); Van Es, et al., Nature 435:959-963 (2005)).

The NOTCH pathway can be activated by four distinct transmembrane receptor types (designated as NOTCH-1?NOTCH-4), which rely on regulated proteolysis. Notch’s expression patterns depend on the cell type. The receptor undergoes sequential cleavage of metalloproteases from the ADAM family after ligand binding (Bru, et. al., Mol. Cell 5:207-216 (2000); Mumm, et al., Mol. Cell 5:197-206 ((2000)) and the presenilin dependent gamma secretase. Selkoe et al. Annu. Rev. Neurosci. 26:565-97 (2003); De Strooper, et al., Nature 398:518-522 (1999)). The last proteolytic cleavage allows the NOTCH receptor’s intracellular domain to be transferred to the cell nucleus, where it interacts to inducing target gene expression.

The NOTCH intracellular domain is subject to ubiquitilation in the cell nucleus. Furin-protease prototeolytic processing and trafficking of NOTCH precursor protein to the cell membrane determine turnover and availability receptors. This, in turn, activates this signaling pathway. Altering glycosylation by Fringe family members of the Notch extracellulardomain may also affect efficiency of ligand binding.

As therapeutic agents for neurological diseases, such as Alzheimer’s disease and leukemia, inhibitors of NOTCH (particularly gamma secretase inhibitors) have been given a lot of attention. These agents could be used to treat or prevent obesity and metabolic syndrome. This would be a significant advance in medicine, public health, and medicine.

The citation or identification of any document within this application does not mean that it is admissible as prior art for the present invention.

The present invention relates to ways of treating or preventing obesity, metabolic disorder, or syndrome in patients by administering a therapeutically-effective amount of a compound which inhibits or modulates NOTCH signaling pathway. Applicants define metabolic syndrome as a modified 2005 National Cholesterol Education Program ATP III definition of 3 or more symptoms. These include: waist circumference?40 in men and?35 in women; blood pressure?130/85 mmHg, triglycerides.150 mg/dL, HDL cholesterol?40mg/dL for men, and 50 mg/dL for women. Glucose?100mg/dL. According to WHO criteria, applicants categorize their body mass index (BMI). BMI 25 kg/m2 = healthy weight, BMI 25-29.9kg/m2=overweight, and BMI?30 kg/m2=obese.

The term “therapeutically effective amount” is used herein. A sufficient amount of a NOTCH inhibit to reduce the body mass index (e.g., 30-36 months) of an obese person to below 30 kg/m2 (preferably below 25 kg/m2), or to increase the weight of an obese individual by at least 15% (and preferably 20, or 25%) The term metabolic syndrome may be used to refer to the amount of NOTCH inhibitor that can reduce or eliminate at least one of these symptoms. It should not be administered more than once per month (e.g., 12 to 36 months). A therapeutically effective dose may be defined as a sufficient amount of the inhibitor that is administered at regular intervals in order to decrease the risk of developing metabolic syndrome or becoming obese by at least 25%. Preferably, it will also reduce the likelihood of patients developing the condition. “Clinically similar” is a term that refers to patients who are clinically similar. The term “clinically similar” may be used to describe patients who aren’t obese and don’t have metabolic syndrome, or patients with similar (e.g. within 15%) NOTCH signaling activities.

NOTCH inhibitor” is a term that may be used to describe an agent capable of blocking NOTCH signaling. An agent that blocks NOTCH signaling may be called a “NOTCH inhibitor”. Mechanisms of action of such NOTCH inhibitors include, but are not limited to, inhibition of gamma-secretase and subsequent suppression of NOTCH receptor cleavage, inhibition of NOTCH trafficking to the cell membrane, suppression of expression or function of ligands and/or receptors, inhibition of ligand turnover, cleavage, and/or endocytosis, modification of NOTCH glycosylation, alteration of ubiquitilation of NOTCH components including the NOTCH intracellular domain, modification of expression and/or activity of co-factors or effectors (e.g., members of the MAML family, RBP-Jkappa/CBF-1), and alteration of differentiation/population of undifferentiated cells in bone marrow, circulating blood, or peripheral tissues (e.g., fat, arteries, veins, heart, heart valves, brain). The preferred inhibitors include receptor antagonists that inhibit the binding of NOTCH molecules to receptors, blocking antibodies against NOTCH component, RNA interfering agent for NOTCH components and small molecules and/or peptides that affect expression, function or activity of NOTCH components. Another approach is to deliver a DNA plasmid that encodes a NOTCH component, or a dominant positive form thereof, locally or systemically. Agents that modulate Notch signaling also include those that can increase expression or activity of Notch components that are involved in the signaling process. Biosimilars can also be included in this category. They inhibit or modulate the expression, function, and activity of ANY NOTCH components.

Another aspect is that levels of NOTCH components expressed, function or activity in biological samples such as peripheral blood can be used to determine if a subject who is not obese, does not have a metabolic disorder, metabolic syndrome or is at higher risk of developing them in the future. Higher levels of NOTCH activity in a patient could indicate an increased risk. The “control group” may be the general population of patients that are not obese or have a metabolic disorder, metabolic syndrome, or metabolic syndrome. The?control group? could be any patient who is not overweight or has a metabolic disorder, metabolic disease or syndrome. Or it may be a general population of patients who are normal and are matched to the patient according to well-known artifacts, such as age, gender, etc. Blood, plasma, serum or urine can be used as the test biological samples. A NOTCH component may be tested for, such as a NOTCH receptor, a NOTCH binding ligand or a Delta3 (Delta?like 3/Dll3) and Delta4 (Delta?like 4/Dll4). Control samples can be chosen using well-known methods and could include blood, serum, or other biological material. Control samples may be taken from people who are healthy and free from cardiovascular disease, or from the general population. These tests can also be used to monitor the effects of new or existing therapies for metabolic syndrome and its complications.

Advanced molecular and functional imaging may allow the visualization of biological or disease processes in vivo. Further, the invention proposes using Notch signaling component expression or activity as imaging biomarkers in order to identify patients with subclinical metabolic diseases and associated diseases. This will allow for the prediction of future development of the metabolic disorder and its complications like atherosclerosis, heart attacks, and aortic narrowing. It also allows for monitoring the effects of therapeutic agents and devices against these diseases. Higher levels of NOTCH activity or expression in a patient may indicate an increased risk.

The invention also includes methods for assaying test compounds to determine if they have potential use in treating or prevention of obesity and metabolic syndrome. This is based on their effect on the NOTCH signaling pathway. Inhibition suggesting potential therapeutic value. To test whether proteolytic cleavage can be prevented, test compounds could be incubated with Gamma-secretase. You can also perform receptor-binding assays using known ligands for NOTCH. For example, Delta1 (Deltalike 1/Dll1) or Delta4 (Deltalike 4/Dll4). Jagged 1 and Jagged 2 may be used (see Ikeuchi, J. Biol. Chem. 278:7751-7754 (2003) to determine the extent of receptor binding. To determine the effect of a test compound, RBP-Jkappa/CBF-1 activity, or expression of known target genes of NOTCH (including the Hes and Hey families) may be used. These assays can be used to identify compounds that may be useful in clinical trials.

It should be noted that terms such as “comprises?”, ‘comprised? or?comprising” are used in this disclosure, and in particular in the claims and/or paragraphs. The meaning of terms like?comprises?, or?comprised? can be attributed to them in U.S. Patent Law. ?consists essentially? They have the same meaning as U.S. Patent Law, e.g. they permit elements not explicitly recited but exclude elements found in prior art or that impact a fundamental or novel characteristic of the invention.

These and other embodiments can be disclosed or made obvious by the following detailed description.

The present invention is partly based on the idea that NOTCH signaling inhibition or modulation can be used to treat obesity and related metabolic disorders as well as the ensuing cardiovascular complications. The therapeutic targets for Notch signaling include ligands like Dll1, Dll3, Dll4, Jagged 1 or Jagged 2, receptors (Notch1, Notch2, Notch3, Notch4, Jagged 1 or Jagged 2), Notch1, Notch2, Notch3, Notch3, Notch4, and all co-factors including Mastermind-like 1, 2, 3 (MAML1-3), and RBP-Jkappa. These methods include but are not limited: systemic or local administrations of blocking antibodies, therapeutics and RNA oligonucleotides (siRNAoligos), antisense and biosimilars.

The present invention relates to therapeutic methods by which an inhibitor or NOTCH signaling is administered in order to treat or prevent obesity and metabolic syndrome. This can be done with any of the NOTCH inhibitors (including gamma secretase inhibitors) that have been described in art. U.S. Patent. Nos. Nos.

The above-described compounds will be given to patients in a combination of a pharmaceutically acceptable carrier and the compound. Any solvent, diluent or liquid vehicle that is pharmaceutically acceptable can be used as a carrier. Standard works of art can provide guidance on how to make pharmaceutical formulations. (See, e.g. Remington’s Pharmaceutical Sciences 16th edition, E. W. Martin Easton, Pa. (1980).) Pharmaceutical compositions can also contain excipients commonly used in the art. Some examples of excipients or carriers that might be present include sugars (e.g. glucose, sucrose), starches such as corn starch, potato starch, and cellulose and its derivatives (e.g. sodium carboxymethylcellulose, ethylcellulose, or celluloseacetate); cellulose and its derivates (e.g. cellulose; malt; gelatin); talc; cocoa butter, oils (e.g. peanut oil; cottonseed oil; sesame oil, soybean oil, oil, oil, oil, vegetable oil; color agents; dispersing agent; coating agents; flavoring; and colouring agents; dispersing; coating agents; flavors; or preservatives

The invention allows for the delivery of compounds via any route that is known to the art. This includes intraoral, internal and rectal routes, nasal, transdermal, intramuscular, intraperitoneal routes, intracutaneous routes, and intracutaneous routes. Drug eluting devices stents are also available as delivery methods. Oral delivery is the preferred method, especially when using tablets, capsules, or solutions. If a compound is sensitive to stomach acid degradation, it can be enterically coated or administered parenterally.

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