Compositions and methods for enhancing intermittent fasting using autophagy inducers

Abstract

The composition contains an autophagy inducer and is administered before, during and / or after an intermittent fasting (IF) diet, preferably a time-restricted feeding (TRF) regimen or an alternate-day fasting (ADF) regimen. For example, the enhancement of intermittent fasting provided by the compositions and methods disclosed herein may include at least one of the following improvements: lifespan, cardiometabolic health, body composition (e.g., reduction in fat mass), cellular senescence (e.g., reduction in inflammatory markers), cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treatment of gastroesophageal reflux disease (GERD). Another aspect is a method of treating or preventing an inflammatory condition, such as rheumatoid arthritis and / or osteoarthritis, by administering an autophagy inducer before, during and / or after an intermittent fasting (IF) diet. An example of an autophagy inducer suitable for the compositions and methods is thymol, optionally in combination with medium-chain triglycerides (MCT).

Description

[Technical field]

[0001]

[0001] The present disclosure generally relates to administering an autophagy inducer before, during and / or after an intermittent fasting (IF) diet, preferably a time-restricted feeding (TRF) regimen or an alternate-day fasting (ADF) regimen. For example, the enhancement of intermittent fasting provided by the compositions and methods disclosed herein may include at least one of improvements in lifespan, cardiometabolic health, body composition (e.g., reduction in fat mass), cellular senescence (e.g., reduction in inflammatory markers), cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treatment of gastroesophageal reflux disease (GERD). [Background technology]

[0002]

[0002] Intermittent fasting (IF) is a generic term used to describe an eating pattern in which periods of little or no absorbed energy intake alternate with periods of feeding (sometimes called feasting).

[0003]

[0003] One of the most common styles of IF is the time-restricted feeding (TRF) regimen, in which food intake is only permitted at specified times per day, for example, people on the 16:8 regimen fast for 16 hours and eat for 8 hours each day. Another common style of IF is alternate-day fasting (ADF), in which 24-hour fasting periods are alternated with 24 hours of ad libitum intake. There are three general types of ADF: (1) total alternate-day fasting, alternating 1 day of eating with 1 day of fasting; no calories are consumed on fasting days, but noncaloric beverages (e.g., water, infusions, tea, coffee) may be consumed; (2) modified alternate-day fasting, alternating 1 day of eating with 1 day of modified fasting, with limited caloric amounts (e.g., ≦500 kcal) of food and beverages consumed on fasting days; and (3) the 5:2 regimen, which involves fasting for 2 days out of 7 days, a modification that allows limited caloric intake on 2 days out of 7 days.

[0004] Based on animal and human studies, IF, when used generally as a weight loss regimen, is also believed to have other health benefits, such as glycemic control, cardiovascular health, and treatment of inflammatory conditions such as rheumatoid arthritis and osteoarthritis (De Cabo, R & Mattson, MP (2019) Effects of Intermittent Fasting on Health, Aging, and Disease. NEJM 381 (26) 2541-2551). The effectiveness of IF on health parameters is highly variable in the literature, most likely due to the diversity of study subjects and study designs. Summary of the Invention

[0005]

[0005] Fasting has been shown to induce autophagy, which is an important mechanism for the longevity and other benefits of intermittent fasting (Longo, VD & Panda, S. (2016) Fasting, circadian rhythms, and time-restricted feeding in healthy lifespan. Cell Metab. 23, 1048-1059). In fact, autophagy is a cellular recycling process involved in cell regeneration.

[0006]

[0006] Existing nutritional solutions target ketosis by MCTs arranged for intermittent fasting. However, to the inventor's knowledge, there is no solution that targets autophagy to enhance the benefits of intermittent fasting. For example, known compositions for intermittent fasting may further include weight loss agents such as MCTs, but the final benefits are typically related to weight loss only. However, the present disclosure provides a combination of autophagy raw material inducers and intermittent fasting regimens to enhance related benefits such as effects on lifespan and cell regeneration.

[0007]

[0007] Thus, the compositions and methods disclosed herein can use an autophagy inducer to induce autophagy in an individual before, during, and / or after intermittent fasting (IF). Preferably, a formulation containing an autophagy inducer is administered to an individual in an amount effective to induce autophagy, for example, in muscle. The formulation can simultaneously promote protein synthesis and removal of damaged cellular material.

[0008]

[0008] In one or more embodiments, the autophagy inducer is selected from the group consisting of thymol, carvacrol, cannabidiolic acid (CBDA), spermidine, urolithin (e.g., urolithin A, urolithin B, or urolithin D), rapamycin, valproic acid, polyphenols (e.g., resveratrol, curcumin), torin-1 (1-[4-(4-propanoylpiperazin-1-yl)-3-(trifluoromethyl)phenyl]-9-quinolin-3-ylbenzo[h][1,6]-naphthyridin-2-one), caffeine, metformin, 5' AMP-activated protein kinase (AMPK) activators, L-type calcium channel inhibitors, ketones (e.g., β-hydroxybutyrate, ketone salts, or ketone ester derivatives), and mixtures thereof.

[0009]

[0009] Non-limiting examples of suitable autophagy inducers include spermidine, palmitic acid, 5-aminoimidazole-4-carboxamide riboside (AICAR), verapamil, nifedipine, diltiazem, piperazine phenothiazine derivatives (e.g., trifluoperazine), ketones (e.g., β-hydroxybutyrate, ketone salts, or ketone ester derivatives), and mixtures thereof. Non-limiting examples of suitable forms of spermidine include spermidine trihydrochloride, spermidine phosphate hexahydrate, spermidine phosphate hexahydrate, and L-arginyl-3,4-spermidine.

[0010] Additionally or alternatively, the autophagy inducer may include one or more autophagy-inducing amino acids, such as glycine, cysteine, proline, glutamic acid (0.1-100 mg), valine, tyrosine, or precursors thereof, such as serine (as a precursor of glycine), N-acetylcysteine ​​(0.1-100 mg), methionine (as a precursor of cysteine), and mixtures thereof. In addition to one or more autophagy-inducing amino acids, optionally, the composition may further include one or more anabolic amino acids, such as leucine, isoleucine, arginine, glutamine, or citrulline. The composition may include one or more anabolic amino acids in an amount effective to activate mTOR in an individual. In some embodiments, the composition includes one or more autophagy-inducing amino acids or precursors thereof in an amount effective to make the composition at least neutral with respect to autophagy, preferably positive with respect to autophagy, even if one or more anabolic amino acids have a negative effect on autophagy.

[0011]

[0011] In some embodiments, autophagy inducers such as thymol, optionally in combination with MCTs, promote satiety; this is supported by Coleman, H., P. Quinn, and MEClegg, "Medium chain triglycerides and conjugated linoleic acids in beverage form increase satiety and reduce food intake in humans," Nutrition Research 36(6):526-33 (2016), and Wymelbeke, VV, J. Louis-Sylvestre, and M. Fantino, "Substrate oxidation and control of food intake in men after a fat-substitute meal compared with meals supplemented with an isoenergetic load of carbohydrate, long-chain triacylglycerols, or medium-chain triacylglycerols," The American Journal of Clinical Nutrition 74(5):620-30 (2001).

[0012]

[0012] Additional features and advantages are described herein, and will be apparent from the drawings and detailed description that follow. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013]

[0013] Definition

[0014] Some definitions are provided below, however definitions may be found in the "Embodiments" section below, and the heading "Definitions" above does not imply that such disclosure in the "Embodiments" section is not a definition.

[0014]

[0015] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated. Thus, for example, reference to "an autophagy inducer" or "the autophagy inducer" encompasses both embodiments having a single autophagy inducer and embodiments having two or more autophagy inducers.

[0015]

[0016] The terms "comprise", "comprises", and "comprising" should be interpreted as inclusive rather than exclusive. Similarly, the terms "include", "including", and "or" should all be interpreted as inclusive unless such interpretation is clearly prevented by the context. However, the compositions disclosed herein may not include elements not specifically disclosed herein. Thus, disclosure of an embodiment using the term "comprising" includes disclosure of an embodiment "consisting essentially of" the specified components, and an embodiment "consisting of" the specified components.

[0016]

[0017] The terms "at least one of" and "and / or" used in the context of "at least one of X or Y" and "X and / or Y", respectively, should be interpreted as "X" or "Y" or "X and Y." For example, "at least one of vitamins or minerals" and "vitamins and / or minerals" should be interpreted as "vitamins without minerals" or "minerals without vitamins" or "both vitamins and minerals."

[0017]

[0018] As used herein, the terms "examples" and "such as," especially when followed by a list of terms, are merely exemplary and illustrative and should not be considered exclusive or inclusive. As used herein, a condition "associated with" or "linked with" another condition means that the conditions occur simultaneously, preferably that the conditions are caused by the same underlying condition, and most preferably that one of the conditions specified is caused by the other condition specified.

[0018]

[0019] Examples of types of time restricted feeding (TRF) include:

[0020] (1) 12:12 12 hours fasting followed by 12 hours of ad libitum feeding;

[0021] (2) 16:8, 16 hours fasting followed by 8 hours of ad libitum feeding; and

[0022] (3) 20:4: 20 hours fasting followed by 4 hours of ad libitum feeding.

[0019]

[0023] There are different types of alternate day fasting (ADF):

[0024] (1) Alternately, one day of feeding is followed by one day of fasting. In this regimen, no calories are consumed on fasting days, but noncaloric beverages (e.g., water, infusions, tea, coffee) may be consumed.

[0025] (2) Modified alternate-day fasting, which alternates between 1 day of feeding and 1 day of modified fasting, with limited caloric amounts (e.g., ≦500 kcal) of food and beverages consumed on the fasting days.

[0026] (3) The 5:2 regimen, which involves fasting for 2 days out of 7. The 5:2 regimen also includes modifications that allow limited caloric intake on 2 days out of 7.

[0020]

[0027] The term "composition" may refer to a food, beverage, dietary supplement, complete nutrition or oral nutritional supplement (ONS) or medical food composition, or mixtures thereof. Within the context of this disclosure, "nutrients" are substances necessary for the health, growth, development and function of an organism, including macronutrients (e.g., proteins, carbohydrates, fats) and their components (e.g., amino acids, sugars, starches, fatty acids, etc.); micronutrients (e.g., vitamins, minerals); other food components (fiber, cholesterol, bioactive phytochemicals, alcohol, etc.); and moisture contained in foods and beverages.

[0021]

[0028] In the context of this disclosure, the terms "food," "food product," and "food composition" refer to products or compositions that are intended for consumption by an individual, such as a human, and that provide nutritional support to an organism, including those that provide energy, nutrients, and water. Compositions of the present disclosure, including many of the embodiments described herein, may comprise, consist of, or consist essentially of one or more autophagy inducers, as well as any additional or optional raw materials or ingredients that are safe for human consumption or otherwise useful in a diet.

[0022]

[0029] The term "Food for Special Medical Purposes (FSMP)" refers to a formula food that is specially processed and prepared to meet the special nutritional or dietary needs of people who suffer from dietary restrictions, digestive and absorption disorders, metabolic disorders, or specific diseases. Such foods are used alone or in combination with other foods under the guidance of a physician or clinical nutritionist. FSMPs are foods for special purposes and are not medicines, but are not usually consumed by healthy people. FSMPs are specially developed by clinicians and nutritionists based on scientific facts from extensive medical research.

[0023]

[0030] The term "oral nutritional supplements (ONS)" refers to sterile liquids, semisolids, or powders that provide macro- and micronutrients. ONS are widely used within emergency and community health settings for individuals who cannot meet their nutritional requirements through oral diet alone.

[0024]

[0031] In the context of this disclosure, the terms "beverage," "beverage product," and "beverage composition" refer to a portable liquid product or composition for consumption by an individual, such as a human, that provides hydration and may also contain one or more nutrients for the individual, and other ingredients that are safe for human consumption.

[0025]

[0032] In the context of this disclosure, a "dietary supplement" is a product taken orally that contains one or more dietary ingredients, such as vitamins, minerals, amino acids, fatty acids, fiber, and / or herbs, and other botanical ingredients used to supplement the diet. Dietary supplements come in many forms and are available as tablets, capsules, powders, liquids, and may be incorporated into certain foods, such as "energy" bars.

[0026]

[0033] As used herein, "complete nutrition" refers to a composition that contains sufficient variety and levels of macronutrients (proteins, lipids, and carbohydrates), micronutrients, and other food components to be the sole source of nutrition for a subject to whom the composition is administered, such that an individual can receive 100% of the individual's nutritional needs from such a complete nutritional composition.

[0027]

[0034] Triglycerides (also known as triacylglycerols or triacylglycerides) are esters derived from glycerol and three fatty acids. The fatty acids can be either unsaturated or saturated. Fatty acids that are not bound to other molecules are called free fatty acids (FFAs).

[0028]

[0035] A medium chain triglyceride (MCT) is a triglyceride in which all three fatty acid moieties in the molecule are medium chain fatty acid moieties. As defined herein, a medium chain fatty acid (MCFA) is a fatty acid having 6 to 14 carbon atoms, preferably 6 to 12 carbon atoms. A medium chain fatty acid having 8 carbon atoms may be referred to herein as a "C8 fatty acid" or "C8". A medium chain fatty acid having 10 carbon atoms may be referred to herein as a "C10 fatty acid" or "C10".

[0029]

[0036] The term "fatty acid moiety" refers to the MCT moiety derived from fatty acid in esterification reaction with glycerol.In a non-limiting example, the esterification reaction between glycerol and only octanoic acid produces MCT containing octanoic acid moiety.In another non-limiting example, the esterification reaction between glycerol and only decanoic acid produces MCT containing decanoic acid moiety.

[0030]

[0037] "Prevention" includes reducing the risk, incidence and / or severity of a condition or disorder. The terms "treatment" and "treating" include both prophylactic or preventative treatment (treatment that prevents and / or delays the onset of the pathological condition or disorder of interest) and curative, therapeutic or disease-modifying treatment, including, for example, therapeutic measures to cure, delay, reduce symptoms and / or halt the progression of a diagnosed pathological condition or disorder, as well as treatment of patients at risk of or suspected of having a disease, and patients who are unwell or diagnosed with a disease or medical condition. The terms "treatment" and "treating" do not necessarily mean that the subject is treated until fully cured. The terms "treatment" and "treating" also refer to maintaining and / or promoting the health of individuals who are not suffering from a disease but who may be susceptible to an unhealthy condition. The terms "treatment" and "treating" are also intended to include the synergism or otherwise potentiation of one or more primary preventative or therapeutic measures. By way of non-limiting example, treatment can be performed by the patient, a caregiver, a doctor, a nurse, or another medical professional.

[0031]

[0038] As used herein, a prophylactically or therapeutically "effective amount" is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or, more generally, reduces symptoms, manages disease progression, or provides a nutritional, physiological, or medical benefit to an individual. The relative terms "improved," "increased," "enhanced," and the like refer to the effect on intermittent fasting by administration of a composition comprising an autophagy inducer compared to an otherwise identical composition that does not contain an autophagy inducer or that contains less of an autophagy inducer. For example, the enhancement of intermittent fasting provided by the compositions and methods disclosed herein can include at least one improvement in lifespan, cardiometabolic health, body composition, cellular senescence, cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treatment of gastroesophageal reflux disease (GERD).

[0032]

[0039] A "subject" or "individual" is a mammal, preferably a human. A "subject" or "individual" may also be an animal. As used herein, the terms "subject" and "individual" are often used to refer to humans, although the present disclosure is not limited to humans.

[0033]

[0040] "Animal" includes, but is not limited to, mammals, including rodents, aquatic mammals, domestic animals such as dogs and cats, livestock such as sheep, pigs, cows and horses, and humans. When "animal", "mammal", or plurals thereof are used, these terms also apply to any animal capable of the effects indicated or intended to be indicated by the context of the text, e.g., animals capable of autophagy.

[0034]

[0041] As used herein, the term "unit dosage form" refers to a physically discrete unit suitable as a dosage unit for human and animal subjects, each unit containing a predetermined amount of a composition disclosed herein, in an amount sufficient to produce the desired effect, together with a pharma- ceutically acceptable diluent, carrier, or vehicle. The specifications for the unit dosage form depend on the particular compound used, the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0035]

[0042] Embodiment

[0043] One aspect of the present disclosure is a method of enhancing an intermittent fasting (IF) diet in an individual before, during, and / or after an IF diet, for example, by improving at least one of lifespan, cardiometabolic health, body composition, cellular senescence, cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treating gastroesophageal reflux disease (GERD). The method includes administering an autophagy inducer (e.g., an effective amount of an autophagy inducer) to an individual (e.g., an individual in need thereof). The autophagy inducer can be administered in a composition that can be administered parenterally, enterally, or intravenously.

[0036]

[0044] Another aspect of the present disclosure is a method of treating, preventing and / or reducing at least one of the risk, occurrence or severity of an inflammatory condition, such as rheumatoid arthritis and osteoarthritis, in an individual before, during and / or after an intermittent fasting (IF) diet. The method includes administering an autophagy inducer (e.g., an effective amount of an autophagy inducer) to an individual (e.g., an individual in need thereof). The autophagy inducer can be administered in a composition that can be administered parenterally, enterally or intravenously.

[0037]

[0045] Preferably, autophagy inducer is orally administered to an individual in a beverage or as a food composition.Non-limiting examples of suitable compositions include food compositions, dietary supplements, dietary supplements (e.g., liquid ONS), complete nutritional compositions, beverages, medicines, oral nutritional supplements, meal replacements, medical foods, nutraceuticals, foods for special medical purposes (FSMP), powdered nutritional formulations that are reconstituted with water or milk before ingestion, food additives, medicines, drinks, pet foods, and combinations thereof.

[0038]

[0046] In one embodiment, the unit dosage form is a predetermined amount of a beverage or food composition (e.g., a predetermined amount of a beverage / food composition comprising an effective amount of an autophagy inducer).

[0039]

[0047] Yet another embodiment is a method of making a composition for administration before, during and / or after an intermittent fasting (IF) diet, comprising adding an autophagy inducer to at least one other ingredient, in some embodiments, the composition is formulated for oral administration and the at least one other ingredient is edible.

[0040]

[0048] In some embodiments, the composition can be a ready-to-drink (RTD) beverage in a container, and the unit dosage form is a predetermined amount of the RTD beverage sealed in the container, and the container is opened for oral administration. For example, the predetermined amount of the RTD beverage can include an effective amount of an autophagy inducer. The RTD beverage is a liquid that can be consumed orally without the addition of further ingredients.

[0041]

[0049] In other embodiments, the method includes forming a beverage by reconstituting a powdered unit dosage form comprising an autophagy inducer in a diluent, such as water or milk, to form a beverage that is subsequently orally administered to an individual (e.g., within about 10 minutes after reconstitution, within about 5 minutes after reconstitution, or within about 1 minute after reconstitution). The powdered unit dosage form can be sealed in a sachet or other package, which can be opened for reconstitution and subsequent oral administration.

[0042]

[0050] The unit dosage form of the dietary supplement may contain excipients, emulsifiers, stabilizers, and mixtures thereof.

[0043]

[0051] In one or more embodiments, the autophagy inducer is selected from the group consisting of thymol, carvacrol, cannabidiolic acid (CBDA), spermidine, a urolithin (e.g., urolithin A, urolithin B, or urolithin D), rapamycin, valproic acid, a polyphenol (e.g., resveratrol, curcumin), torin-1 (1-[4-(4-propanoylpiperazin-1-yl)-3-(trifluoromethyl)phenyl]-9-quinolin-3-ylbenzo[h][1,6]-naphthyridin-2-one), caffeine, metformin, a 5′ AMP-activated protein kinase (AMPK) activator, an L-type calcium channel inhibitor, a ketone (e.g., β-hydroxybutyrate, a ketone salt, or a ketone ester derivative), and mixtures thereof.

[0044]

[0052] In some embodiments, about 1 mg to about 1 g, preferably about 10 mg to 500 mg, more preferably about 20 mg to 200 mg of thymol and / or carvacrol per daily dose is administered to an individual in one or more divided doses. In another embodiment, about 120 mg of thymol and / or carvacrol per daily dose is administered to an individual in one or more divided doses. Thymol (10-64%) is one of the main constituents of the essential oil of thyme (Thymus vulgaris L, Lamiaceae). Carvacrol is present in the essential oil of oregano (Origanum vulgare), thyme oil, oil obtained from cress, and monarda. The essential oil of thyme subspecies contains 5% to 75% carvacrol, whereas the Savory (Satureja) subspecies has a content of 1% to 45%. Origanum majorana and Dittany of Crete are rich in carvacrol, 50% and 60-80%, respectively. Some embodiments of the present compositions therefore include plants and / or concentrated plant extracts, essential oils or fractions, particularly from thyme and oregano, that provide at least a portion of the thymol and / or carvacrol in the composition.

[0045]

[0053] Further non-limiting examples of suitable autophagy inducers are urolithins, spermidine, palmitic acid, 5-aminoimidazole-4-carboxamide riboside (AICAR), verapamil, nifedipine, diltiazem, piperazine phenothiazine derivatives (e.g., trifluoperazine), ketones (e.g., β-hydroxybutyrate, ketone salts, or ketone ester derivatives), and mixtures thereof. Non-limiting examples of suitable forms of spermidine include spermidine trihydrochloride, spermidine phosphate hexahydrate, spermidine phosphate hexahydrate, and L-arginyl-3,4-spermidine.

[0046]

[0054] The method may include the step of daily administering an autophagy inducer (e.g., spermidine) in a weight range of 0.05 mg to 1 g per kg of body weight, preferably 1 mg to 200 mg per kg of body weight, more preferably 5 mg to 150 mg per kg of body weight, even more preferably 10 mg to 120 mg per kg of body weight, or most preferably 40 mg to 80 mg per kg of body weight.

[0047]

[0055] In some embodiments, the urolithin may be administered in an amount of about 0.2 to 150 milligrams (mg) of urolithin per kilogram (kg) of the subject's body weight. Preferably, the urolithin is administered at a dose equal to or corresponding to 2 to 120 mg of urolithin per kg of the subject's body weight, more preferably 4 to 90 mg of urolithin per kg of the subject's body weight, and most preferably 8 to 30 mg of urolithin per kg of the subject's body weight.

[0048]

[0056] Additionally or alternatively, the autophagy inducer can include one or more autophagy-inducing amino acids, such as glycine, cysteine, proline, glutamic acid, valine, tyrosine, or precursors thereof, such as serine (as precursor of glycine), N-acetylcysteine, methionine (as precursor of cysteine), and mixtures thereof. In one embodiment, glycine can be administered in an amount of about 0.1-100 milligrams (mg) of glycine or functional derivatives thereof per kilogram (kg) of the subject's body weight. A daily dose of the composition can include one or more of the following: 0.1-100 mg glycine per kg of body weight (bw); 5-340 mg valine per kg of body weight; 20-126 mg tyrosine per kg of body weight; 3-43 mg methionine per kg of body weight. The composition includes one or more autophagy-inducing amino acids or precursors thereof in an amount effective to render the composition at least neutral with respect to autophagy, preferably positive with respect to autophagy.

[0049]

[0057] In addition to one or more autophagy-inducing amino acids, the composition may optionally further comprise one or more anabolic amino acids, such as leucine, isoleucine, arginine, glutamine or citrulline. A daily dose of the composition may comprise one or more of the following: 0.175-142.85 mg leucine per kg body weight, preferably 0.35-71.425 mg leucine per kg body weight; 0.175-71.425 mg isoleucine per kg body weight; 20-300 mg arginine per kg body weight, preferably 50-200 mg arginine per kg body weight, and / or 20-300 mg citrulline per kg body weight, preferably 100-200 mg citrulline per kg body weight. A daily dose of one or more anabolic amino acids may be provided by a single or multiple servings of the composition per day.

[0050]

[0058] The composition can include one or more anabolic amino acids in an amount effective to activate mTOR in an individual. In some embodiments, the composition includes one or more autophagy-inducing amino acids or precursors thereof in an amount effective to render the composition at least neutral with respect to autophagy, and preferably positive with respect to autophagy, in the event of a negative effect on autophagy from one or more anabolic amino acids.

[0051]

[0059] In some embodiments, the method includes administering a composition disclosed by any of U.S. Patent Application Publication No. 16 / 954,694, entitled "Compositions and Methods Using a Combination of Autophagy Inducer and High Protein for Induction of Autophagy," WO 2020 / 245299, entitled "Compositions and Methods Using One or More Autophagy-Inducing Amino Acids to Potentiate Musculoskeletal Effect of One or More Anabolic Amino Acids," or WO 2020 / 254663, entitled "Compositions and Methods Using Thymol and / or Carvacrol For Induction of Autophagy," each of which is incorporated by reference in its entirety.

[0052]

[0060] In some embodiments, the composition is administered less than one month before the implementation of an IF regimen, for example, less than one week before the implementation of an IF regimen. Additionally or alternatively, the composition is administered at least the majority of the days during the implementation of an IF regimen, for example, every day during the implementation of an IF regimen. Additionally or alternatively, the composition is administered at least one week after the implementation of an IF regimen, for example, at least one month after the implementation of an IF regimen.

[0053]

[0061] In certain embodiments, the method increases plasma spermidine concentration in a subject before, during, and / or after an IF regimen, for example, to a concentration in the range of 50-6000 nmol / L plasma, preferably 100-6000 nmol / L plasma. The method may include administering a daily dose of an autophagy inducer (e.g., spermidine) in the weight range of 0.05 mg / kg to 1 g / kg body weight, preferably 1 mg / kg to 200 mg / kg body weight, more preferably 5 mg / kg to 150 mg / kg body weight, even more preferably 10 mg / kg to 120 mg / kg body weight, or most preferably 40 mg / kg to 80 mg / kg body weight.

[0054]

[0062] Typically, a daily dose of 50 pg to 10 g of an autophagy inducer, preferably a spermidine compound, is administered to a subject in one or more divided doses before, during, and / or after an IF regimen.

[0055]

[0063] Wheat germ is rich in spermidine, and therefore some embodiments of the present compositions include wheat germ and / or concentrated wheat germ extract, which provides at least a portion of the autophagy inducers in the composition.

[0056]

[0064] Whey protein is rich in BCAAs such as leucine and isoleucine.Therefore, some embodiments of the composition include whey protein, which provides at least a portion of the anabolic amino acids in the composition.Furthermore, hydrolyzed collagen is a rich source of glycine and proline, and therefore some embodiments of the composition include hydrolyzed collagen, which provides at least a portion of the autophagy-inducing amino acids in the composition.

[0057]

[0065] In certain non-limiting embodiments, the composition comprises an autophagy inducer (preferably at least one of thymol oil or oregano oil), MCT (preferably MCT oil), and optionally at least one additional ingredient selected from the group consisting of a protein (e.g., one or more of collagen, pea protein), a gum (e.g., xanthan), one or more vitamins (e.g., at least one of vitamin B12, vitamin B1, or vitamin D), one or more minerals (e.g., at least one of iron, zinc, or magnesium), natural flavors, natural colors, and mixtures thereof.

[0058]

[0066] As used herein, the term "protein" includes amino acids in free form, molecules of 2-20 amino acids (herein referred to as "peptides"), as well as longer chains of amino acids. Small peptides, i.e., chains of 2-10 amino acids, are suitable for the present compositions, either alone or in combination with other proteins. "Free form" amino acids are the monomeric forms of amino acids. Suitable amino acids include both natural and unnatural amino acids. The present compositions may include a mixture of one or more proteins, e.g., a mixture of one or more of (i) peptides, (ii) longer chains of amino acids, or (iii) free form amino acids; the mixture is preferably formulated to obtain a desired amino acid profile / content.

[0059]

[0067] At least a portion of the protein may be of animal or plant origin, for example a milk protein, e.g., a dairy protein, such as one or more of a milk protein concentrate or milk protein isolate; a caseinate or casein, e.g., a casein micelle concentrate or casein micelle isolate; or a whey protein, e.g., a whey protein concentrate or whey protein isolate. Additionally or alternatively, at least a portion of the protein may be a vegetable protein, such as one or more of a soy protein or a pea protein.

[0060]

[0068] Mixtures of these proteins are also suitable, such as mixtures in which casein is the majority but not all of the protein, whey protein is the majority but not all of the protein, pea protein is the majority but not all of the protein, and soy protein is the majority but not all of the protein. In one embodiment, at least 10% by weight of the protein is whey protein, preferably at least 20% by weight, more preferably at least 30% by weight. In one embodiment, at least 10% by weight of the protein is casein, preferably at least 20% by weight, more preferably at least 30% by weight. In one embodiment, at least 10% by weight of the protein is vegetable protein, preferably at least 20% by weight, more preferably at least 30% by weight.

[0061]

[0069] The whey protein can be, for example, any whey protein selected from the group consisting of whey protein concentrate, whey protein isolate, whey protein micelles, whey protein hydrolysate, acid whey, sweet whey, denatured sweet whey (sweet whey from which the caseinoglycomacropeptide has been removed), a fraction of whey protein, and any combination thereof.

[0062]

[0070] The casein may be obtained from any mammal, but is preferably obtained from cow's milk, and preferably as casein micelles.

[0063]

[0071] The proteins may be non-hydrolyzed, partially hydrolyzed (i.e., peptides with molecular weights between 3 kDa and 10 kDa and an average molecular weight less than 5 kDa), or extensively hydrolyzed (i.e., 90% of the peptides have a molecular weight less than 3 kDa), for example, in the range of 5% to 95% hydrolyzed. In some embodiments, the peptide profile of the hydrolyzed protein may be in different molecular weight ranges. For example, the majority of the peptides (greater than 50 mol% or greater than 50 wt%) may have a molecular weight within 1-5 kDa, or 5-10 kDa, or 10-20 kDa.

[0064]

[0072] In one embodiment, the composition comprises a carbohydrate source. Any suitable carbohydrate may be used in the composition, including, but not limited to, starch (e.g., modified starch, amylose starch, tapioca starch, corn starch), sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, xylitol, sorbitol, or combinations thereof.

[0065]

[0073] The carbohydrate source preferably represents no more than 50% of the energy of the composition, more preferably no more than 36% of the energy of the composition, and most preferably no more than 30% of the energy of the composition.

[0066]

[0074] In one embodiment, the composition comprises a fat source. The fat source may comprise any suitable fat or fat mixture. Non-limiting examples of suitable fat sources include vegetable fats, such as olive oil, corn oil, sunflower oil, high oleic sunflower oil, rapeseed oil, canola oil, hazelnut oil, soybean oil, palm oil, coconut oil, black currant seed oil, borage oil, lecithin, and the like, animal fats, such as milk fat, and the like, or combinations thereof. The composition comprising an autophagy inducer (e.g., spermidine) can be administered to an individual in a therapeutically effective dose before, during, and / or after intermittent fasting (IF). The therapeutically effective dose can be determined by one skilled in the art and may vary depending on a number of factors known to those skilled in the art, such as the severity of the condition and the weight and general condition of the individual.

[0067]

[0075] The composition is preferably administered to an individual at least 2 days per week, more preferably at least 3 days per week, most preferably 7 days per week; at least 1 week, at least 1 month, at least 2 months, at least 3 months, at least 6 months, or even longer. In some embodiments, the composition is administered to an individual continuously for several days, for example, at least until a therapeutic effect is achieved. In some embodiments, the composition can be administered to an individual daily for at least 30, 60, or 90 consecutive days.

[0068]

[0076] The above dosing examples do not require uninterrupted daily administration. Rather, there may be several short interruptions in administration, such as a 2-4 day break during the administration period. The ideal duration of administration of the composition can be determined by one of skill in the art.

[0069]

[0077] In a preferred embodiment, the composition is administered orally or enterally (e.g., by tube feeding) to an individual. For example, the composition can be administered to an individual as a drink, capsule, tablet, powder, or suspension.

[0070]

[0078] The composition may be any type of composition suitable for human and / or animal consumption. For example, the composition may be selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional formulations that are reconstituted with water or milk before ingestion, food additives, medicines, beverages and drinks. In one embodiment, the composition is an oral nutritional supplement (ONS), a complete nutritional formula, a medicine, a drug or a food product. In a preferred embodiment, the composition is administered to an individual as a drink. The composition may be stored as a powder in a sachet and then suspended in a liquid such as water for use.

[0071]

[0079] In certain non-limiting examples, preferred forms of the composition are a beverage (e.g., comprising water and / or another liquid), a ready-to-drink beverage (e.g., comprising water and / or coffee) that does not require the addition of any other ingredients prior to ingestion; an oral nutritional supplement (ONS); or a coffee creamer.

[0072]

[0080] In some cases where oral or enteral administration is not possible or recommended, the composition may also be administered parenterally. A preferred parenteral administration is intravenous administration. A particular form of parenteral administration is intravenous delivery of nutrients. Parenteral nutrition is "total parenteral nutrition" when food is not provided by other routes. "Parenteral nutrition" is preferably an isotonic or hypertonic aqueous solution (or a solid composition that is dissolved, or a liquid concentrate that is diluted to obtain an isotonic or hypertonic solution) that contains sugars such as glucose, and further contains one or more of lipids, amino acids, and vitamins.

[0073]

[0081] In some embodiments, the compositions are administered to an individual in a single dosage form, i.e., all compounds are present in one formulation given to the individual with a meal. In other embodiments, the compositions are co-administered in separate dosage forms, e.g., at least one component is co-administered separately from one or more of the other components of the composition.

[0074]

[0082] In view of the disclosure herein, an embodiment provided by the present disclosure is a composition comprising an autophagy inducer and formulated for administration to an individual before, during and / or after an intermittent fasting (IF) regimen, preferably a time restricted feeding (TRF) regimen or an alternate day fasting (ADF) regimen, the composition preferably further comprising medium chain triglycerides (MCTs) and optionally at least one additional ingredient selected from the group consisting of proteins, gums, vitamins, minerals, natural flavors, and natural colors.

[0075]

[0083] Preferably, the autophagy inducer comprises at least one of thymol, preferably at least partially provided by thyme oil or oregano oil in the composition, or carvacrol, preferably at least partially provided by oregano oil in the composition. In some embodiments, the composition comprises a protein, preferably at least one of collagen, pea protein). In some embodiments, the composition comprises a gum, preferably xanthan. In some embodiments, the composition comprises a vitamin, preferably at least one of vitamin B1, vitamin B12, or vitamin D. In some embodiments, the composition comprises a mineral, preferably at least one of iron, zinc, or magnesium.

[0076]

[0084] In some embodiments, the autophagy inducer is selected from the group consisting of thymol, carvacrol, cannabidiolic acid (CBDA), spermidine, urolithin, rapamycin, valproic acid, polyphenols, caffeine, metformin, ketones, palmitic acid, 1-[4-(4-propanoylpiperazin-1-yl)-3-(trifluoromethyl)phenyl]-9-quinolin-3-ylbenzo[h][1,6]-naphthyridin-2-one, 5' AMP-activated protein kinase (AMPK) activators, L-type calcium channel inhibitors, 5-aminoimidazole-4-carboxamide riboside (AICAR), verapamil, nifedipine, diltiazem, piperazine phenothiazine derivatives, one or more autophagy-inducing amino acids, and mixtures thereof.

[0077]

[0085] Preferably, the composition has a form selected from the group consisting of a beverage, an oral nutritional supplement (ONS), and a coffee creamer. The composition may also be for animal consumption.

[0078]

[0086] Compositions intended for non-human animals include food compositions, animal treats (e.g., biscuits), and / or dietary supplements that supplement the nutritional needs of animals. The compositions may be dry compositions (e.g., kibble), semi-moist compositions, wet compositions, or any mixture thereof. In one embodiment, the composition is a dietary supplement, such as a gravy, drinking water, beverage, yogurt, powder, granule, paste, suspension, chew, morsel, treat, snack, pellet, pill, capsule, tablet, or other suitable delivery form. In one embodiment, another suitable delivery form includes encapsulation of at least one of the active ingredients of the composition by macrocapsule or microcapsule filling. Microcapsule filling includes, for example, microcapsules, microparticles, microspheres, and microemulsions. Techniques of macrocapsule filling and microcapsule filling, including chemical, physicochemical, or mechanical methods, are well known in the art.

[0079]

[0087] Dietary supplements may need to be or can be mixed with water or other diluents prior to administration to the animal.

[0080]

[0088] Another embodiment is a unit dosage form of any of the compositions disclosed herein, the unit dosage form comprising an autophagy inducer in an amount effective to enhance an intermittent fasting (IF) diet of an individual to whom the unit dosage form is administered. Preferably, the enhancement of the IF diet comprises at least one of improvement in lifespan, cardiometabolic health, body composition, cellular senescence, cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treatment of gastroesophageal reflux disease (GERD).

[0081]

[0089] Another embodiment is a method of enhancing an intermittent fasting (IF) diet, comprising administering any of the compositions disclosed herein to a subject before, during and / or after the IF diet. Preferably, the composition is administered to the subject daily for at least one week.

[0082]

[0090] Another embodiment is a method of making a composition for administration before, during and / or after an intermittent fasting (IF) diet, comprising adding an autophagy inducer to at least one other ingredient selected from the group consisting of medium chain triglycerides (MCT), protein, gum, vitamins, minerals, natural flavors, and natural colors. EXAMPLES

[0083]

[0091]

[0092] The following non-limiting examples are representative of one or more embodiments disclosed herein.

[0084]

[0093] Helminth survival assay on solid agar plates

[0094] C. elegans strains were cultivated at 20°C on nematode growth medium agar plates seeded with E. coli strain OP50. Thymol was dissolved in DMSO and added at the indicated concentrations just before pouring the plates. Worms were exposed to compounds throughout their lifespan from egg to death. Plates were changed twice a week to ensure constant exposure to compounds. Control populations were treated with the corresponding 1% concentration of DMSO. For lifespan measurements, parental F0 L4 worms were grown until they reached adulthood and laid eggs on treated plates. Thus, the derived F1 worms were exposed to compounds throughout their lifespan from egg to death. Adults were manually scored daily as dead or alive. Worms were considered dead when they were not moving spontaneously and did not respond to touching of the head. Control populations were treated with the corresponding 1% concentration of DMSO.

[0085]

[0095] Helminth survival assay in microfluidics

[0096] A worm suspension is first injected into the microfluidic device. The geometry of the device is optimized to retain only adult worms in the worm culture chamber by simply selecting an appropriate flow rate for sample injection. After isolating a defined worm population in the chamber, the worms are cultured and treated on-chip with thymol at a controlled temperature of 20°C. The worm culture is maintained in the chamber by perfusion with E. coli. The microfluidic chip is integrated into an inverted microscope combined with a camera that continuously records videos (every 10 hours). The images are longitudinally analyzed by various parameters such as body bends frequency, speed, curvature and amplitude readouts of the movements at the head, tail and mid-body, automatically providing information about the health span and vitality of the worms.

[0086]

[0098] It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present subject matter and without diminishing its intended advantages. Accordingly, such changes and modifications are intended to be encompassed by the appended claims. [Brief description of the drawings]

[0087] [Figure 1] 4 shows that treatment of worms with thymol at a concentration of 50 μM on solid agar plates has no significant effect on lifespan (n=60 worms / group). [Diagram 2] Figure 1 shows that thymol at a concentration of 50 μM extends lifespan when worms are treated in microfluidics. P values ​​represent comparison to vehicle calculated using the log-rank test. Thymol alone (control) does not statistically improve lifespan, and the effect on lifespan is statistically significant when the worms are under conditions mimicking IF.

Claims

1. A composition comprising an autophagy inducer and formulated for administration to an individual before, during and / or after an intermittent fasting (IF) regimen.

2. 2. The composition of claim 1, wherein the autophagy inducer comprises at least one of thymol; thymol provided at least in part by thyme and oregano essential oils in the composition; carvacrol; or carvacrol provided at least in part by oregano oil in the composition.

3. The composition of claim 1 or 2, wherein the composition further comprises medium chain triglycerides (MCTs).

4. 10. The composition of claim 1, wherein the composition comprises at least one additional ingredient selected from the group consisting of proteins, gums, vitamins, minerals, natural flavors, and natural colors.

5. 10. The composition of claim 1, wherein the composition comprises at least one of vitamin B1, vitamin B12, or vitamin D.

6. The composition of claim 1 , wherein the composition comprises at least one of iron, zinc, or magnesium.

7. 2. The composition of claim 1, wherein the autophagy inducer is selected from the group consisting of thymol, carvacrol, cannabidiolic acid (CBDA), spermidine, urolithin, rapamycin, valproic acid, polyphenols, caffeine, metformin, 1-[4-(4-propanoylpiperazin-1-yl)-3-(trifluoromethyl)phenyl]-9-quinolin-3-ylbenzo[h][1,6]-naphthyridin-2-one, 5′ AMP-activated protein kinase (AMPK) activators, L-type calcium channel inhibitors, ketones, palmitic acid, 5-aminoimidazole-4-carboxamide riboside (AICAR), verapamil, nifedipine, diltiazem, piperazine phenothiazine derivatives, one or more autophagy-inducing amino acids, and mixtures thereof.

8. 10. The composition of claim 1, in a form selected from the group consisting of meal replacements, dietary supplements, complete nutritional compositions, beverages, pharmaceuticals, oral nutritional supplements, medical foods, nutraceuticals, foods for special medical purposes (FSMP), powdered nutritional formulations to be reconstituted with water or milk before ingestion, food additives, pharmaceuticals, drinks, pet foods, and combinations thereof.

9. 9. The composition of claim 8, wherein the dietary supplement is a gravy, drinking water, beverage, yogurt, powder, granules, paste, suspension, chew, bite, treat, snack, pellet, pill, capsule, tablet, macrocapsule fill, or microcapsule fill.

10. 10. A unit dosage form of the composition of claim 1, comprising an amount of the autophagy inducer effective to enhance an intermittent fasting (IF) diet in an individual to whom the unit dosage form is administered.

11. 11. The unit dosage form of claim 10, wherein the enhancement to an intermittent fasting (IF) diet comprises at least one improvement in longevity, cardiometabolic health, body composition, cellular senescence, cellular regeneration, ketosis, weight loss, glycemic control, blood pressure, satiety, or treatment of gastroesophageal reflux disease (GERD).

12. 1. A method of making a composition for administration before, during, and / or after an intermittent fasting (IF) diet, comprising adding an autophagy inducer to at least one other ingredient selected from the group consisting of medium chain triglycerides (MCTs), proteins, gums, vitamins, minerals, natural flavors, and natural colors.

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