Compounds for cancer cachexia

Abstract

The present invention provides a compound or a pharmaceutically acceptable salt or solvate thereof for treating cancer cachexia in a subject, wherein the compound has the general formula I: (I); wherein -R1 is a C1-C6 alkyl group with up to five -OR2 substituents, wherein R2 represents hydrogen or a C1-C6 alkyl group, or wherein -OR2 represents a (R)-3-hydroxybutyrate moiety; or -R1 is a moiety derived from an alcohol HOR1, wherein the alcohol is a sugar. In general, the present invention also provides treatments for conditions associated with cachexia, such as muscle atrophy. #imgabs0#

Description

Field of the Invention

[0001] The present invention relates to compounds for preventing or treating cachexia, particularly cachexia associated with cancer, in a subject. Background of the Invention

[0003] Cancer cachexia is a wasting syndrome suffered by cancer patients, characterized by weight loss, anorexia, fatigue and anemia. It causes involuntary muscle loss in patients. Cachexia adversely affects the patient's ability to resist infection and withstand chemotherapy and radiotherapy treatment. As a result, the patient's body begins to lose weight.

[0004] A reduction in food intake (to <1500 kcal / day), along with a 10% or greater weight loss and systemic inflammation, are considered prognostic parameters for cancer cachexia. Weight loss cannot be reversed with nutritional supplements. Ultimately, cancer cachexia leads to death, with patients dying when total body weight loss reaches 25-30%.

[0005] The mechanisms involved in cancer cachexia appear to be complex and multifactorial. Current treatment modalities include appetite stimulants and drugs targeting cachexia signaling molecules and mediators, which help prevent and treat wasting.

[0006] As discussed in previous patent US9,801,903, ketogenic diets are well known and have been widely used to treat cancer. In general, ketogenic diets are characterized by high fat content, moderate to low protein content, and very low carbohydrate content. The use of ketogenic diets in cancer has shown potentially promising but inconsistent effects (Oliveira et al.; J Acad Nutr Diet, 2017 "Anutritional perspective of ketogenic diet incancer: A narrative review."). The use of ketogenic diets is based on the basic theory of metabolic therapy targeting abnormal energy metabolism of cancer cells. The ketogenic state limits glucose utilization and destroys glycolysis in cancer cells, while providing alternative energy sources for healthy cells. This selectively starves cancer cells, while normal cells are not damaged.

[0007] Woolf et al. in Front Mol Neurosci, November 2016, Vol. 9, No. 122, “Tumour metabolism, the ketogenic diet and β-hydroxybutyrate: Novel approaches to adjuvant brain tumour therapy” disclose that the ketogenic diet may be a promising anti-cancer therapy. The paper discusses how the ketogenic diet has been shown to reduce angiogenesis, inflammation, peritumoral edema, and tumor migration and invasion. In addition, the diet can enhance the effects of radiation and chemotherapy in a mouse glioma model, thereby improving survival. In addition, the paper discusses how ketone bodies themselves have anti-tumor effects, so ketone supplements can be effective against certain diseases when used alone.

[0008] Attempts have been made to reverse cachexia and selectively deprive tumors of energy-generating metabolic substrates by feeding a ketogenic diet. For example, Tisdale et al., Br J Cancer (1987) Vol. 56, pp. 39-43, attempted to reverse cachexia using a diet based on metabolic differences between tumor and host tissues, with the goal of selectively feeding the host by consuming the tumor.

[0009] GB2517088A discloses the use of ketone body esters in the treatment of muscle breakdown. Cachexia is mentioned in this publication, but cancer cachexia is not specifically disclosed.

[0010] There are conflicting references in the literature regarding whether cancer patients with cachexia should be placed on a ketogenic diet. Tumor cells are known to have high rates of glucose consumption, show increased rates of aerobic glycolysis, and are susceptible to carbohydrate deprivation. In addition, many tumors lack certain key mitochondrial enzymes and thus have greatly lost the ability to utilize fat or ketone bodies for energy production. Therefore, it is thought that replacing glucose with ketone bodies reduces the energy supply to the tumor while maintaining the energy supply to the host. On the other hand, there is some concern that cancer patients may lose too much weight on a ketogenic diet.

[0011] It is generally understood that the term "ketone bodies" includes three compounds: D-β-hydroxybutyrate, acetoacetate, and acetone. D-β-hydroxybutyrate is also known as βHB or (R)-3-hydroxybutyrate. During periods of low food intake, the liver produces ketone bodies from fatty acids.

[0012] WO2004 / 108740 discloses nutritional supplements and therapeutic compositions containing (R)-3-hydroxybutyric acid derivatives. The compositions disclosed therein can be used to treat tumors, in particular brain tumors, such as astrocytomas. There is no mention of the treatment of cancer cachexia.

[0013] Shukla et al. disclosed the use of ketone salts - sodium hydroxybutyrate and lithium acetoacetate to treat pancreatic cancer cachexia in Cancer Metab (2014) Vol. 2, p. 18, "Metabolic reprogramminginduced by ketone bodies diminishes pancreatic cancer cachexia". Reduced tumor growth and inhibition of muscle and weight loss were shown. Large amounts of ketone salts were used, which would be dangerous if used in vivo.

[0014] Administration of ketone salts cannot be used to increase βHB or AcAc concentrations to appropriate levels for humans (e.g., concentrations in the range of 10 to 20 mM in the blood) because such salt levels may cause acidosis, hypertension, and / or renal failure. It is not possible for humans to ingest the required amount of salt without greatly exceeding the recommended daily salt dosage. Gastrointestinal discomfort and hypertension are often the result of high salt intake.

[0015] Therefore, there is a need for new effective treatments for treating cancer cachexia in a subject. SUMMARY OF THE INVENTION

[0017] Therefore, in a first aspect, the present invention provides a compound for preventing or treating cancer cachexia in a subject, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound has the general formula I:

[0018]

[0019] in

[0020] - R1 is C1-C6 alkyl, said alkyl carrying up to five -OR2 substituents, wherein R2 represents hydrogen or C1-C6 alkyl, or wherein -OR2 represents a (R)-3-hydroxybutyrate moiety; or

[0021] -R1 is a moiety derived from an alcohol HOR1, wherein the alcohol is a sugar.

[0022] In a second aspect, the present invention provides a compound for treating cancer in a subject, which is (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester of the following formula:

[0023]

[0024] In a third aspect, the present invention provides a compound for use in treating cancer in a subject, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound has the general formula I:

[0025]

[0026] in

[0027] - R1 is C1-C6 alkyl, said alkyl carrying up to five -OR2 substituents, wherein R2 represents hydrogen or C1-C6 alkyl, or wherein -OR2 represents a (R)-3-hydroxybutyrate moiety; or

[0028] - R1 is a moiety derived from an alcohol HOR1, wherein the alcohol is a sugar;

[0029] Wherein, one hour after administration of the compound, the subject's blood (R)-3-hydroxybutyrate concentration is in the range of 1-20 mM.

[0030] Other embodiments of the present invention include:

[0031] A method of treating cancer cachexia in a subject, comprising administering to said subject a therapeutically effective amount of a compound of formula I as defined above.

[0032] A method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of (R)-1,3-butylene glycol mono-3-hydroxybutyrate of the formula:

[0033]

[0034] A method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound of formula I as defined above, wherein one hour after administration of the compound, the subject's blood (R)-3-hydroxybutyrate level is in the range of 1-20 mM.

[0035] Use of a compound of formula I as defined above in the preparation of a medicament for treating cancer cachexia in a subject.

[0036] Use of (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester of the formula in the preparation of a medicament for treating cancer in a subject

[0037]

[0038] Use of a compound of formula I as defined above in the preparation of a medicament for treating cancer in a subject, wherein one hour after administration of the compound to the subject, the subject's blood (R)-3-hydroxybutyric acid level is in the range of 1-20 mM. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 Survival time of mice is shown - (A) survival curves of all five experimental groups; (B) main effect of exercise status; (C) main effect of nutritional status;

[0041] Figure 2 (A) Food intake and (B) body weight of different experimental groups are shown;

[0042] Figure 3 Blood parameters of different experimental groups are shown—(A) blood βHB and (B) blood glucose levels;

[0043] Figure 4 The wheel activity is shown;

[0044] Figure 5 Tumor weights are shown;

[0045] Figure 6 The muscle weights of different experimental groups are shown - (A) tibialis anterior (TA), (B) quadriceps femoris (QUAD), and (C) gastrocnemius (GAS) weights;

[0046] Figure 7 Grip strength of the different experimental groups on day 16 is shown - (A) forelimb and (B) hindlimb grip strength; and

[0047] Figure 8 Shown are the muscle contraction properties of the soleus muscle measured in vitro - peak twitch (A) and tonicity (B) of the soleus muscle - for different experimental groups. DETAILED DESCRIPTION OF THE INVENTION

[0049] The compounds of the present invention provide a source of (R)-3-hydroxybutyric acid in a subject. Therefore, the compounds are esters of (R)-3-hydroxybutyric acid, which can be decomposed by esterases in the body to form (R)-3-hydroxybutyric acid.

[0050] (R)-3-Hydroxybutyrate is a ketone body as defined in "Metabolic Regulation: A Human Perspective" by KN Frayn.

[0051] WO2004 / 108740 discloses that ketone bodies can be directly administered to a subject to achieve elevated ketone body levels. However, in some cases direct administration of ketone salts or acids can be difficult and risky, so the use of esters has been proposed as a preferred alternative. The preparation of ketoesters has been disclosed, for example, in WO2014 / 140308, which describes a method for preparing (R)-3-hydroxybutyl 3-hydroxybutyrate.

[0052] Esters of (R)-3-hydroxybutyric acid can be prepared by transesterification of ethyl (R)-3-hydroxybutyrate with an alcohol. The reaction can be enzyme catalyzed. For example, ethyl (R)-3-hydroxybutyrate and (R)-1,3-butanediol can be reacted together in the presence of an immobilized lipase under moderate vacuum to remove the resulting ethanol byproduct.

[0053] In a first embodiment of the present invention, the ester of (R)-3-hydroxybutyric acid is a compound of formula I:

[0054]

[0055] in

[0056] - R1 is a C1-C6 alkyl group, said alkyl group having up to five -OR2 substituents;

[0057] - wherein R2 represents hydrogen or C1-C6 alkyl, or wherein -OR2 represents a (R)-3-hydroxybutyrate moiety; or

[0058] -R1 is a moiety derived from an alcohol HOR1, wherein the alcohol is a sugar.

[0059] Typically, zero, one or two -OR2 groups represent a (R)-3-hydroxybutyrate moiety. Preferably, only zero or one -OR2 group represents a (R)-3-hydroxybutyrate moiety.

[0060] The R1 moiety is derived from the corresponding alcohol HO-R1. The alcohol HO-R1 can be, for example, a monohydric alcohol, a dihydric alcohol, a polyhydric alcohol or a sugar.

[0061] Preferably, in Formula I, R1 is C1-C6 alkyl substituted by 0, 1, 2, 3, 4 or 5 -OR2 substituents. Most preferably, R1 is C1-C6 alkyl substituted by 1, 2 or 3 -OR2 substituents, typically 1 or 2 -OR2 substituents.

[0062] Preferably, R2 is H.

[0063] Preferably, R1 has the formula –CH2-CH(OH)-CH2(OH) or –CH2-CH2-CH(OH)-CH3. In these cases, R1 is a moiety derived from an alcohol HO-R1, which corresponds to butanediol and glycerol, respectively. The butanediol may be racemic 1,3-butanediol. Most preferably, the alcohol HO-R1 corresponds to R-1,3-butanediol. In this case, the group R1 has the following formula:

[0064]

[0065] Preferably, the compound used in the first aspect of the invention is a monoester, i.e., in the case where the alcohol HO-R1 contains more than one pendant hydroxyl group, only one of these hydroxyl groups reacts to form a hydroxybutyrate moiety. Partial esters are compounds in which the alcohol HO-R1 contains more than one pendant hydroxyl group and not all of these hydroxyl groups react to form a hydroxybutyrate moiety.

[0066] We have found that (R)-3-hydroxybutyric acid (R)-1,3-butylene glycol monoester and (R)-3-hydroxybutyric acid partial glycerol ester provide high circulating levels of (R)-3-hydroxybutyric acid in the blood. In addition, these esters provide surprisingly high levels of uptake in the intestine, thereby enabling high blood concentrations of (R)-3-hydroxybutyric acid to be achieved when the beverage is consumed.

[0067] Thus, in a preferred embodiment, the present invention provides hydroxybutyrate esters or partial esters, such as (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester and (R)-3-hydroxybutyric acid glycerol partial ester for use in treating cancer cachexia in a subject.

[0068] (R)-3-hydroxybutyl 3-hydroxybutyrate is particularly advantageous because it allows a substantial increase in blood (R)-3-hydroxybutyrate to be achieved by oral ingestion of a smaller volume of the substance than the racemic ketone. Subjects who ingest the substance are more likely to ingest enough ketones to provide a physiologically beneficial response without the risk of physical discomfort (e.g., due to ingestion of large amounts of fluid or bitter / other objectionable tastes). (R)-3-hydroxybutyl 3-hydroxybutyrate also elevates (R)-3-hydroxybutyrate concentrations for a longer period of time than ketone salts. A lower dosage frequency is then required to maintain higher (R)-3-hydroxybutyrate levels. This also facilitates subject compliance with the dosing regimen.

[0069] Therefore, a particularly preferred compound for use in the first aspect of the present invention is (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester, also known as (R)-3-hydroxybutyl (R)-3-hydroxybutyrate, of the formula:

[0070]

[0071] Another preferred compound of the present invention is (R)-3-hydroxybutyric acid-glycerol partial ester, ie (R)-3-hydroxybutyric acid-glycerol monoester or diester.

[0072] In various embodiments of the present invention, R1 is derived from an alcohol HOR1, wherein the alcohol is a sugar. The sugar may be selected from altrose, arabinose, dextrose, erythrose, fructose, galactose, glucose, gulose, idose, lactose, lyxose, mannose, ribose, ribulose, sucrose, talose, threose and xylose.

[0073] In the case where R1 is derived from an alcohol HOR1 which is a polyol, the polyol may be chosen from glycerol, sorbitol and xylitol.

[0074] When the compounds of the present invention contain chiral centers other than those depicted in the above formula, the compounds may exist as racemic mixtures or as pure enantiomers.

[0075] The compounds of the present invention may be present as physiologically compatible salts. For example, sodium, potassium, calcium or magnesium salts thereof may be used.

[0076] Administration of the compounds defined above to a subject can treat or prevent cachexia in a subject, particularly cachexia in a patient suffering from cancer. As described above, cachexia is a wasting syndrome suffered by a patient characterized by an involuntary loss of muscle mass. "Cancer cachexia" is a wasting syndrome experienced by patients suffering from cancer.

[0077] In the treatment of cancer cachexia, the muscle mass of the subject can be maintained or increased. For example, the compound or composition defined herein can be administered to the subject over a period of time, wherein the muscle mass increases by at least 1 percentage point, preferably by at least 3 percentage points, desirably by at least 5 percentage points. Typically, the compound or composition is administered to the subject at least once a day, for example 1, 2 or 3 times a day. Typically, the composition is administered to the subject during and after its cancer treatment.

[0078] Preferably, during the intake period of the present invention, the total body weight of the subject is maintained or increased. Typically, in the present invention, the normal food intake of the subject is maintained or increased.

[0079] Suitably, the compound or composition defined herein, preferably (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester, is ingested at a level of at least 100 mg ketone per kg body weight per day. Desirably, the compound is ingested at a level sufficient to provide a plasma ketone ((R)-3-hydroxybutyric acid) concentration of at least 0.1 mM, preferably at least 0.2 mM, more preferably at least 1 mM and optimally at least 2 mM. Typically, within one hour of administering the compound, the plasma level of (R)-3-hydroxybutyric acid is within the range of 1-20 mM. Suitably, the compound or composition is ingested at a level such that the blood ketone concentration does not exceed 20 mM, suitably does not exceed 10 mM or 8 mM and may not exceed 5 mM.

[0080] The blood concentration of the ketone will depend on the body weight of the individual, and we have found that oral administration of at least 300 mg of (R)-1,3-butanediol monoester of (R)-3-hydroxybutyric acid per kg body weight provides a plasma concentration of about 1.5 mM (R)-3-hydroxybutyric acid, and administration at 500 mg / kg provides a plasma concentration of at least 3 mM (R)-3-hydroxybutyric acid. At a dose of 700 mg / kg subject body weight, the blood (R)-3-hydroxybutyric acid concentration is suitably at least 5 mM, preferably 6 mM. When 1 g / kg subject body weight of the monoester is administered orally, the blood (R)-3-hydroxybutyric acid concentration is suitably at least 7 mM, preferably at least 8 mM, especially at least 9 mM. The dosing regimen includes multiple drinks taken separately.

[0081] Blood levels of (R)-3-hydroxybutyrate can be determined using commercially available test kits, for example, (R)-3-hydroxybutyrate can be measured in whole blood using a handheld monitor and reagent strips (Precision Xtra, Abbott Diabetes Care, UK).

[0082] The present invention provides for the treatment or prevention of cachexia-related disorders, such as muscle wasting, weight loss, and loss of appetite.

[0083] In another embodiment of the invention, administration of a compound or composition as defined herein increases the contractile properties of the subject's muscles. This may advantageously result in an increase in the subject's strength.

[0084] In another embodiment of the invention, during treatment of cachexia, the subject's activity level may be increased. Thus, during treatment, the subject may feel more motivated to engage in physical activity and become more active.

[0085] In the treatment of cachexia, the survival of the subject can be improved. Thus, the subject can live longer than without treatment.

[0086] In the treatment of cachexia, tumor growth can be inhibited.

[0087] Muscle mass, the contractile properties of muscle, the activity level of a subject, tumor growth and survival of a subject can be measured in mammals, such as mice, by the methods used in the Examples, and in humans using bioimpedance, body calipers, activity monitors (e.g., pedometers), dual energy X-ray (DEXA) scans, CT scans, and MR scans, among others.

[0088] As used herein, the term cancer refers to a physiological condition that is typically characterized by unregulated cell growth, i.e., a proliferative disease. Examples of such proliferative diseases include carcinomas, lymphomas, blastomas, sarcomas, and leukemias.

[0089] In particular, administration can treat cachexia associated with colon cancer.

[0090] The compounds defined herein may be administered together with a carbohydrate. The carbohydrate may be, for example, a monosaccharide such as glucose, fructose or maltose. The compounds may alternatively (or simultaneously) be administered together with a protein or an amino acid.

[0091] In another embodiment of the invention, the compound (R)-(R)-1,3-butanediol mono-3-hydroxybutyrate, also known as (R)-3-hydroxybutyl 3-hydroxybutyrate, is administered to a subject in a method of treating cancer.

[0092] The subject can be an animal, preferably a mammal, such as a human.

[0093] In one embodiment, the compound, e.g., (R)-3-hydroxybutyl 3-hydroxybutyrate, is not used to treat liver cancer. For example, the cancer is not hepatocellular carcinoma. More particularly, the compound may not be used to reduce liver fat in a subject with hepatocellular carcinoma.

[0094] The compounds of the present invention may not be used to treat subjects with fatty liver disease, including non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis (ASH) and non-alcoholic fatty liver (NAFL), cirrhosis and hepatocellular carcinoma. For example, the compounds may not be used to reduce liver fat in these subjects.

[0095] Preferably, the cancer is colon cancer.

[0096] As described above, the subject may be suffering from cancer cachexia.

[0097] The compound for the present invention can be included in the nutritional composition. Suitably, the nutritional composition comprises water and a source of (R)-3-hydroxybutyric acid. Preferably, the composition comprises an ester of (R)-3-hydroxybutyric acid, a flavoring agent, and optionally one or more of protein, carbohydrates, sugars, fats, fibers, vitamins and minerals. Suitably, the flavoring agent can include a fruit-based flavoring agent. In one embodiment, the flavoring agent is suitably bitter, such as coffee, chocolate and cranberry. Bitter flavoring agents can be combined with other flavoring agents, such as fruit-based flavoring agents, such as flavoring agents based on grapefruit, raspberries and cranberries.

[0098] The composition of the present invention may contain a mixture of isomers of the compound of formula I above.

[0099] The composition is suitably organoleptically acceptable. "Organoleptically acceptable" means that the composition must have acceptable organoleptic properties of taste, color, feel and odor.

[0100] The composition may comprise medium chain triglycerides (MCT). If present, the medium chain triglycerides preferably comprise a compound having the formula CH2R a -CH2R b -CH2R c Medium chain triglycerides, of which R a , R b and R c is a fatty acid having 5 to 12 carbon atoms. Suitably, R a , R b and Rc It is a fatty acid containing a six-carbon backbone (tri-C6:0) because tri-C6:0 MCT is reported to be rapidly absorbed by the gastrointestinal tract.

[0101] The composition of the present invention may include a derivative of L-carnitine or L-carnitine. Examples of derivatives of L-carnitine include decanoylcarnitine, hexanoylcarnitine, caproylcarnitine, lauroylcarnitine, octanoylcarnitine, stearoylcarnitine, myristoylcarnitine, acetyl-L-carnitine, O-acetyl-L-carnitine and palmitoyl-L-carnitine. When carnitine is used, the composition of the present invention suitably includes: i) ketone bodies, preferably ketone monoesters, more preferably (R)-3-hydroxybutyric acid monoesters, and ii) derivatives of L-carnitine or L-carnitine, and optional MCT.

[0102] Where MCT and L-carnitine or a derivative thereof are used, it is appropriate to emulsify the MCT with carnitine. Preferably, 10 to 500 g of emulsified MCT is combined with 10 to 2000 mg of carnitine, for example 50 g of MCT (95% tri-C8:0) is emulsified with 50 g of mono- and di-glycerides combined with 500 mg of L-carnitine. Preferably, the level of the (R)-3-hydroxybutyrate source is greater than that of the MCT.

[0103] The composition according to the invention may be provided in any suitable form, including solids, such as powders, tablets, bars, confectionery products or granules, liquids, such as beverages, gels, capsules or any other conventional product forms. The composition may be a food, a food supplement, a dietary supplement, a functional food or a nutraceutical or a component thereof.

[0104] Examples of foods to which the composition can be incorporated as an additive include snack bars, cereals, candies, and probiotic preparations, including yogurts. Examples of beverages include soft drinks, alcoholic beverages, energy drinks, dry drink mixes, nutritional drinks, and herbal teas for infusion or herbal mixtures for decoction in water.

[0105] The compounds used in the present invention are typically formulated as nutraceuticals.

[0106] When in solid form, the composition suitably comprises at least 5% by weight of the compound of the invention, more preferably at least 10% by weight and up to 95% by weight of the composition. Although, for example, when the composition is a dry powder intended for use with a liquid to prepare a liquid composition, a content of 15 to 30% by weight of the dry composition may be suitable, a solid stick or product form suitably comprises 30 to 95% by weight of the composition. In particular, 50-95% by weight.

[0107] When the composition is in solid form, the composition may further comprise one or more of the following components:

[0108] - diluents, such as lactose, dextrose, sucrose, cellulose, corn starch or potato starch;

[0109] - lubricants, for example silicon dioxide, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol;

[0110] - binders, for example starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone;

[0111] - disintegrants, for example starch, alginic acid, alginates or sodium starch glycolate;

[0112] - Effervescent agents;

[0113] - colorants;

[0114] - flavoring agents;

[0115] - Wetting agents, such as lecithin, polysorbates, lauryl sulfate; and / or

[0116] -Carrier.

[0117] Where the composition is in liquid form, the composition suitably comprises a compound as defined herein in an amount of at least 1 wt %, for example 3 to 40 wt % of the liquid composition, but may be higher, for example up to 50 wt % of the composition, depending on whether the composition is intended to be taken in a single dose or in a number of smaller doses to achieve the desired blood ketone level.

[0118] The composition in liquid form may comprise several liquid components suitably blended together, or may comprise liquid and solid components suitably mixed with or dissolved in liquid components. In one embodiment, the dry composition comprising the compound defined above is diluted with a suitable liquid such as water, juice, yogurt or milk, preferably, the ratio of dry composition to liquid is 1:1 to 1:10, more preferably 1:3 to 1:7.

[0119] The composition can be provided as required in the form of a liquid product in an easy-to-take form or in the form of a concentrate or paste suitable for dilution at the time of use. The diluent for use with the liquid composition is preferably milk, juice or water.

[0120] If desired, the composition may also be provided in encapsulated form, provided that the encapsulating material and the amount used are suitable for safe human consumption.

[0121] One aspect of the invention provides a compound of the invention as defined above in a pharmaceutical composition, together with one or more pharmaceutically acceptable excipients.

[0122] The compound used in the present invention can exist in the form of a pharmaceutically acceptable salt. As used herein, a pharmaceutically acceptable salt is a salt formed with a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid, and organic acids, such as citric acid, fumaric acid, maleic acid, malic acid, ascorbic acid, succinic acid, tartaric acid, benzoic acid, acetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid. Pharmaceutically acceptable bases include alkali metals (such as sodium or potassium) and alkaline earth metals (such as calcium or magnesium) hydroxides and organic bases, such as alkylamines, arylalkylamines and heterocyclic amines.

[0123] The compounds used in the present invention may exist in the form of solvates. The term "solvate" refers to a complex or aggregate formed by one or more solute molecules, i.e., a compound of the present invention or a pharmaceutically acceptable salt thereof, and one or more solvent molecules. Such solvates are typically crystalline solids with a substantially fixed molar ratio of solute to solvent. Representative solvents include, for example, water, methanol, ethanol, isopropanol, acetic acid, etc. When the solvent is water, the solvate formed is a hydrate.

[0124] The compounds of the present invention contain chiral centers. Therefore, they can be used in the form of racemic mixtures, enantiomers or mixtures enriched in one or more stereoisomers. The scope of the present invention as described and claimed includes the racemic forms of the compounds of the present invention as well as individual enantiomers and mixtures enriched in stereoisomers.

[0125] It should be understood that the term "or a pharmaceutically acceptable salt or solvate thereof" is intended to include all permutations of salts and solvates, such as solvates of pharmaceutically acceptable salts of the compounds of the present invention.

[0126] The pharmaceutical composition comprises a compound of the invention mixed with one or more pharmaceutically acceptable diluents, excipients or carriers. Even though the compounds useful in the invention (including pharmaceutically acceptable salts, esters and pharmaceutically acceptable solvates thereof) can be administered alone, they are usually administered in admixture with a pharmaceutical carrier, excipient or diluent, particularly for human treatment. The pharmaceutical composition can be used for human or animal use in human and veterinary medicine.

[0127] Examples of such suitable excipients for use in the various forms of pharmaceutical compositions described herein can be found in "Handbook of Pharmaceutical Excipients, 2nd Edition, (1994)", edited by A Wade and PJ Weller.

[0128] (Pharmaceuticals and Nutrition) The composition may include a pharmaceutically acceptable adsorbent. Suitably, the adsorbent adsorbs the compound of the present invention in or on the adsorbent. Advantageously, the user experiences a lower degree of the taste of the compound (which may taste offensive) compared to the taste experienced by taking the same composition without an absorbent. Preferably, the adsorbent includes a lattice or void capable of retaining the compound of the present invention. Any adsorbent used in food or known for food can be used. Examples of suitable adsorbents include polymer hydrogels, such as cross-linked polycarboxylate homopolymers or copolymers, inclusion compounds, cyclic oligosaccharides, such as cyclodextrins, and milk powder. The adsorbent may be present in any desired content according to the particular formulation, and may be 5% to 80% by weight of the composition, such as 10% to 50% by weight.

[0129] Typically, the use of the invention involves oral, parenteral or intravenous administration of the compound. Oral administration is preferred.

[0130] The present invention also provides a compound as defined herein, in substantially pure form or in association with one or more pharmaceutically acceptable diluents or carriers, for use in treating cancer, particularly cancer cachexia, in a subject.

[0131] As used herein, the term "substantially pure form" generally refers to a compound having a purity of 50% or more, preferably 75% or more, more preferably 90% or more, even more preferably 95% or more, and most preferably 99% or more.

[0132] The following examples illustrate the invention. Example

[0133] General Materials and Methods

[0134] Animal Housing and Housing - Male Balb / c mice were obtained at 11 weeks of age and housed in polycarbonate cages at controlled room temperature (21-23°C) and humidity (40-60%) with a 6:00 AM to 6:00 PM light cycle. Mice were housed individually to allow assessment of individual energy intake and physical activity parameters. Cages contained spruce pellets ( BK 8 / 15) pads were changed every three weeks. Enrichment was provided by two cotton balls, which were replaced at cage change. Mice received a wooden stick to enable them to properly grind their teeth when receiving a liquid diet. Solid food was provided in a recess in the wire lid on the top of the cage, while mice were fed via a feeding tube (Bio-Serv TM , catalog number 14-726-518) were provided with a liquid diet daily. Water was provided by a centralized system with a metal dispenser extending into the cage. All procedures were approved by the Animal Ethics Committee of the University of Leuven, Belgium (P093 / 2017).

[0135] Experimental diets - Powdered Lieber-DeCarli diets were purchased from Bio-Serv (products F1258SP and F1259SP; Bio-Serv, Frenchtown, NJ, USA). The control diet (CON) was prepared by adding warm water to the Lieber-DeCarli control diet powder (F1259SP). The ketone ester diet (KET) was prepared by adding warm water, (R)-3-hydroxybutyl 3-hydroxybutyrate (ketoester), and maltodextrin (Canderel, Ternat, Belgium) to the F1258SP powder. Commercially available vanilla flavoring (0.4% v / v; Dr. Oetker, Diegem, Belgium) was added to both liquid diets to increase palatability. The macronutrient composition of the diets is listed in Table 1. The energy (kcal), protein, and fat per gram of food were equal for both diets.

[0136] Table 1: Macronutrient composition of experimental diets

[0137]

[0138] General Study Protocol and Dietary Intervention - Before the start of the study, mice were housed individually and received standard chow ad libitum ( R / MH, Soest, Germany). Food intake was recorded and body weight was measured daily for 5 days to assess their normal food intake rate. After this baseline follow-up period, mice were matched for body weight and food intake and then randomly assigned to five experimental groups: i) quiescent C26 tumor-bearing mice receiving a control diet (CON 静止 ), ii) quiescent C26 tumor-bearing mice on a ketogenic diet (KET 静止 ), iii) C26 tumor-bearing mice on a voluntary wheel (WR) receiving a control diet (CON ex ), iv) C26 tumor-bearing mice undergoing WR on a ketogenic diet (KET ex), and v) quiescent healthy mice (WT) receiving a control diet. The mice were habituated to their experimental diet by gradually changing their food supply. During the first three days, the mice received an experimental liquid diet (without added ketones) as well as standard chow. From days 3 to 6, the mice received only their experimental liquid diet (i.e., 0% KET or CON diet). Thereafter, ketone esters were added to the diet in the KET group to produce a concentration of 3% on day 6 and 4.5% on day 9. Three matched (CON 静止 , KET 静止 and WT) and a matching pair (CON ex and KET ex CON and WT mice were given a diet that provided the same energy content (kcal) as that ingested by their pair-fed KET mice the previous day.

[0139] Tumor induction and exercise intervention - On day 12, 5x10 50 μL of phosphate-buffered saline (PBS) was injected interscapularly. 5 Tumor xenografts were induced by inoculating mouse colon cancer C26 cells. The C26 cell line was cultured in DMEM (Thermo Fisher Scientific, 41965-062) supplemented with 10% fetal bovine serum (FBS), 1% glutamine (Sigma G7513) and 1% penicillin and streptomycin. WT mice were injected with the same volume of PBS. As previously reported in Coletti D et al., WR mice (CON) were injected with 10% fetal bovine serum (FBS), 1% glutamine (Sigma G7513) and 1% penicillin and streptomycin. ex and KET ex ) received a running wheel to induce voluntary exercise. Substrains of inbred mice differ in their physical activity as a function of behavior. Sci World J 2013:1–7, 2013. Each wheel was equipped with a commercial cycle computer (2Cycle, 81488) to record physical activity parameters (i.e., running time, distance, and speed).

[0140] Specific experimental methods and procedures

[0141] Survival Experiment - Forty mice were randomly assigned to different experimental groups according to their initial body weight and treated as described above. Mice were allowed to survive beyond their expected lifespan and were euthanized only when considered moribund. Body weight, food intake, and wheel running parameters were measured daily. No additional measurements were performed to avoid confounding effects on survival time.

[0142] Behavioral Testing and Tissue Collection Experiments - In the second experiment, 9 mice per experimental group were used for behavioral testing and tissue collection. Mice were euthanized 16 days after PBS or tumor cell inoculation. The running wheels were removed twenty-four hours before sacrifice to eliminate potential acute effects of exercise on the measured variables.

[0143] Body weight, food intake, and wheel-running parameters - Body weight was measured every 3 days during food training. After tumor induction, body weight was measured every 4 days. Food intake and wheel-running parameters were measured daily.

[0144] Grip Strength - Grip strength of the forelimbs and hindlimbs was assessed at baseline (day 5) and every 4 days from tumor induction by a bar and a grid connected to an isometric force transducer (BIO-GS3 Grip Strength Test, BIOSEB, Pinellas Park, Florida). The mice were raised by their tails and positioned to grasp the metal bar or grid and then pulled horizontally until they could not maintain their grip. The maximum force (g) was recorded in five attempts for the forelimbs and hindlimbs. Performance was calculated as the average of the three best attempts. Between attempts, the mice were returned to their cages for a 3-minute rest period.

[0145] Blood metabolite concentrations - Blood parameters were measured 1 hour postprandially, every 3 days during diet adaptation, and every 4 days from the day of tumor induction. Blood βHB and glucose levels were measured using a Glucomen Lx plus instrument (Menarini Diagnostics, Firenze, Italy) with Lxβ-ketone or Lx glucose strips, respectively. The detection limit of β-hydroxybutyrate was 0.1 to 8.0 millimolar (mM). Blood lactate concentration was assessed using the Lactate-Pro 2 monitoring system (Arkray, Kyoto, Japan). Measurements were performed according to the manufacturer's instructions.

[0146] Muscle contractile properties - Isolated muscle contractile properties were assessed as described in Deravé W et al.; Skeletal muscle properties in a transgenic mouse model for amyotrophic lateral sclerosis: Effects of creatine treatment. Neurobiol Dis 13:264–272, 2003 and Eijnde BO et al.; Effect of muscle creatine content manipulation on contractile properties in mouse muscles. Muscle and Nerve 29:428–435, 2004.

[0147] Mice were anesthetized by intraperitoneal injection of ketamine-xylazine (10 mg / kg). After dissection of the soleus and extensor digitorum longus (EDL) of both hind limbs, the muscles were vertically fixed on a force transducer (HSE, March-Hugstetten, Germany) and incubated in an organ bath containing Krebs-Henseleit solution (118 mM NaCl, 25 mM NaHCO3, 5 mM KCl, 1 mM MgSO4, 1 mM KH2PO4, 2.5 mM CaCl2 and 1 mM glucose). The organ bath was continuously aerated with a mixture of 95% O2 and 5% CO2 and maintained at 25°C. After fixation, a 15-minute stabilization period was allowed to recover from the dissection procedure. After recovery, the resting length (L0) was determined using a micropositioner and oscilloscope by tetanically stimulating the soleus and EDL at 80 Hz and 120 Hz, respectively, for 350 ms and interspersed with 2 minutes of rest intervals. The muscles were then stimulated to assess twitch and rigidity characteristics. The muscles were electrically stimulated 3 times at 5 Hz, interspersed with a 1-minute rest interval (twitch stimulation), 500 ms (soleus) at 80 Hz and 250 ms (EDL) at 100 Hz, interspersed with a 2-minute rest interval (rigidity stimulation). The average maximum tension and specific tension (force corrected for muscle cross-sectional area) of twitch and rigidity stimulation were calculated. The wet muscle mass was divided by the resting muscle length (L0) and the mammalian skeletal muscle density (1.06 g / cm 3 ) to determine the cross-sectional area.

[0148] Statistical analysis-Data are expressed as mean ± SEM. Two-way analysis of variance (two-way ANOVA) was used, followed by Tukey's multiple comparison test to determine which groups had significant differences to evaluate the differences between experimental groups. Mice from the WT group were not included in the two-way ANOVA model, but were added as reference. Unpaired t-tests were used to assess differences in wheel activity. For survival experiments, the Kaplan-Meier method was used to estimate the curves for each experimental group, and the log-rank (Mantel-Cox) test and Tukey's test were used to test the differences in survival time, so that the I-type error rate judgment family (family-wise Type I error rate) was maintained at 0.05. Statistical significance was defined as p<0.05.

[0149] result

[0150] Figure 1 Shows the survival time – Figure 1 (A) includes the survival curves of all five experimental groups; Figure 1 (B) shows the main effect of exercise condition (CON休息 and KET 休息 Relative to CON ex and KET ex ); Figure 1 (C) shows the main effect of nutritional status (CON 休息 +CON WR Relative to KET 休息 +KET ex ). The keto group mice showed a significant increase in both mean and median survival times compared to the control group (+40% and +17%, respectively, p<0.01). Survival was similar between the rest and exercise groups.

[0151] Figure 2 Food intake and body weight of different experimental groups are shown - (A) food intake and (B) body weight. ex and KET ex ) had a higher average food intake than the rest group (CON 休息 and KET 休息 ) was about 15% higher in the KET and CON groups, while no difference was observed between the KET and CON groups. In all experimental groups, body weight decreased similarly over time.

[0152] Figure 3 Blood parameters of the different experimental groups are shown - (A) blood βHB and (B) blood glucose levels. At baseline, blood βHB levels were undetectable in all groups (<0.1 mM). During the dietary intervention, βHB levels in the KET group increased up to about 2 mM, while no increase above baseline was found in CON. Blood glucose levels were consistently lower in KET compared to the CON group.

[0153] Figure 4 The wheel activity is shown. ex , compared to KET ex Mice in the 50% 1% 2% 2% 3% 4% 5% 2% 1% 2% 3% 1% 2% 3% 2% 3% 4% 5% 2% 3% 4 ...

[0154] Table 2: Wheel activity in mice when fed a control diet or a ketone ester diet

[0155]

[0156] Figure 5 Tumor weights are shown. Tumor weights in KET mice were significantly lower (204±32 mg, p<0.05) compared to CON group (455±22 mg). Tumor weights were similar between rest and exercise groups.

[0157] Figure 6The muscle weights of the different experimental groups are shown - (A) tibialis anterior (TA), (B) quadriceps femoris (QUAD), and (C) gastrocnemius (GAS) weights. The TA weight was on average about 12% higher in KET (41.6 ± 2.1 mg, p < 0.05) compared to CON (37.0 ± 1.5 mg). The QUAD weight was lower in the exercise group than in the rest group (44.4 ± 1.1 vs. 51.4 ± 1.6 mg, p < 0.05). The GAS weight was similar between the two groups.

[0158] Figure 7 The grip strength of the different experimental groups on day 16 is shown - (A) forelimb and (B) hindlimb grip strength. Ketone group (KET 休息 and KET ex ) had higher grip strength in both forelimb and hindlimb than the control group (CON 休息 and CON ex ,p<0.05). Exercise itself did not change grip strength.

[0159] Figure 8 Shown are the muscle contraction properties of the soleus muscle measured in vitro - peak twitch (A) and tonicity (B) of the soleus muscle in different experimental groups. 休息 and KET ex ) were higher than those of the control group (CON 休息 and CON ex , p<0.05) were about 45% and about 29% higher. Exercise itself did not change muscle strength.

Claims

1. Use of a compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating cancer cachexia in a subject, wherein the normal food intake of the subject is maintained, wherein the compound is (R)-3-hydroxybutyric acid (R)-1,3-butanediol monoester having the following formula:

2. The use of a compound according to claim 1, wherein the compound is not administered together with medium chain triglycerides.

3. The use of a compound according to claim 1 or 2, wherein the compound is administered to a subject in a therapeutically effective amount, wherein one hour after administration of the compound, the blood (R)-3-hydroxybutyrate level in the subject is in the range of 1-20 mM.

4. Use of the compound according to claim 1 or 2, wherein the cancer is colon cancer.

5. The use of the compound according to claim 1 or 2, wherein the compound is also used to inhibit tumor growth.

6. The use of a compound according to claim 1 or 2, wherein the compound is also used to improve the survival of a subject.

7. The use of the compound according to claim 1 or 2, wherein the compound is also used to maintain or increase muscle mass in the subject.

8. The use of a compound according to claim 1 or 2, wherein the compound is also used to maintain or increase the activity level of the subject.

9. Use of the compound according to claim 1 or 2, which is used together with a carbohydrate.

10. The use of the compound according to claim 9, wherein the carbohydrate is a monosaccharide or maltose.

11. Use of a compound according to claim 10, wherein the monosaccharide is glucose or fructose.

12. Use of a compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating cancer cachexia in a subject, wherein the compound is (R)-1,3-butanediol mono(R)-3-hydroxybutyrate having the following formula: wherein the compound is administered to a subject in a therapeutically effective amount, wherein one hour after administration of the compound, blood (R)-3-hydroxybutyrate levels in the subject are in the range of 1-20 mM.

13. Use of a pharmaceutical composition comprising the compound as defined in any preceding claim and one or more pharmaceutically acceptable excipients for the preparation of a medicament for preventing or treating cancer cachexia in a subject.

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