Medium-chain fatty acid ester of butanediol, composition, oral preparation and application method thereof

Abstract

Aspects of the present disclosure include medium-chain fatty acid esters of butanediol, compositions thereof, oral formulations thereof, and methods of use thereof.

Description

[0001] This application is a divisional application. The international application number of the original application is PCT / US2017 / 035826, the international application date is June 2, 2017, the Chinese national application number is 201780041428.2, the entry date into the Chinese national phase is January 2, 2019, and the name of the invention is "Medium-chain fatty acid esters of β-hydroxybutyrate and butanediol and compositions and methods of use thereof".

[0002] Statement Regarding Federally Funded Research

[0003] This invention was made with government support under Grant Nos. R24 DK085610 and K08 AG048354 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS

[0005] This application claims priority under 35 USC §119(e) to the filing date of U.S. Provisional Patent Application No. 62 / 346,975, filed on June 7, 2016, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0006] The present disclosure relates to compounds including medium-chain fatty acid esters of butanediol, compositions and oral preparations containing the compounds, and uses of the compounds. Background Art

[0007] Ketogenic diets and ketone bodies are of interest for treating a variety of human diseases, including epilepsy, dementia, and diseases of aging. Ketone bodies are small compounds produced from fat that can replace sugar when the body's energy reserves are depleted, such as during fasting or intense exercise. Ketogenic diets stimulate ketone body production by containing very small amounts of sugar or other carbohydrates. The main ketone bodies in the human body are acetoacetate (AcAc) and beta-hydroxybutyrate (BHB). Ketogenic diets are used clinically as a treatment for epilepsy, but they are often difficult to stick to for long periods of time. The extremely high fat content (and low carbohydrate content) can make foods on a ketogenic diet unpalatable and sometimes cause gastrointestinal problems, kidney stones, high cholesterol, and other side effects.

[0008] BHB is a metabolic intermediate that is the currency for generating cellular energy but also has several signaling functions separate from energy production. Either or both of the energy and signaling functions may be important for the effects of BHB on human disease. During periods when glucose is scarce, such as during fasting or intense exercise, BHB is the currency for the conversion of energy stored in adipose tissue into fuel, which can be used by cells throughout the body to maintain their function. Fat mobilized from adipose tissue is transported to the liver and converted into BHB. BHB circulates in the blood to all tissues. After being taken up by cells, BHB is broken down in the mitochondria, producing acetyl-CoA, which is further metabolized into ATP. This is the canonical "energy currency" function of BHB.

[0009] Furthermore, BHB may have a variety of signaling functions. Most of these are unrelated to its function as an energy currency, as they are effects of the BHB molecule itself and are generally secondary to its metabolism to acetyl-CoA and ATP. Signaling functions may include: 1) inhibition of class I and IIa histone deacetylases, resulting in changes in histone modification and gene expression, as well as changes in acetylation status and protein activity of non-histone proteins; 2) metabolism to acetyl-CoA resulting in increased cellular production of acetyl-CoA, which serves as a substrate for acetyltransferases, resulting in similar changes in protein acetylation of histones and non-histone proteins, such as deacetylase inhibition; 3) covalent attachment to histones and possibly other proteins in the form of lysine-β-hydroxybutyrylation, which may have similar effects to lysine-acetylation; 4) binding and activation of the hydroxycarboxylic acid receptor 2 (HCAR2) receptor, resulting in changes in adipose tissue metabolism; 5) binding and inhibition of the free fatty acid receptor 3 (FFAR3) receptor, resulting in activation of the sympathetic nervous system and changes in systemic metabolic rate; 6) inhibition of the NOD-like receptor 3 (NLRP3) inflammasome. Summary of the invention

[0010] Aspects of the present disclosure include fatty acid β-hydroxy ester compounds (e.g., fatty acid esters of β-hydroxybutyrate), fatty acid esters of butanediol, and pharmaceutically acceptable salts thereof. Pharmaceutical compositions having one or more fatty acid β-hydroxy ester compounds and / or one or more fatty acid esters of butanediol are also provided. Methods of treating a subject by administering one or more esters to the subject are also provided. Kits containing one or more subject esters are also described. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1a and 1b Depicts the C6 substituted esters of β-hydroxybutyrate according to certain embodiments. 1 H-NMR and GC-MS.

[0012] Figure 2a and 2b The C8 substituted esters of β-hydroxybutyrate according to certain embodiments are respectively depicted. 1 H-NMR and GC-MS.

[0013] Figure 3a and 3b The C6 substituted fatty acid β-hydroxy ester compounds according to certain embodiments are respectively depicted. 1 H-NMR and GC-MS.

[0014] Figure 4a and 4b The C6 acyl substituted esters of butanediol according to certain embodiments are respectively depicted. 1 H-NMR and GC-MS.

[0015] Figure 5a and 5b The C8 acyl substituted esters of butanediol according to certain embodiments are respectively depicted. 1 H-NMR and GC-MS.

[0016] Figure 6a-6c The biological functions of C6 and C8 esters of butanediol and β-hydroxybutyrate were depicted as tested by intraperitoneal injection into wild-type C57BL / 6 male mice at two doses each. Figure 6a Plasma concentrations within 6 hours after injection of C6 and C8 esters of β-hydroxybutyrate are depicted. Figure 6b Plasma concentrations of C6 and C8 esters of butanediol are depicted over a 6-hour period following injection. Figure 6c Plasma concentrations of C6 and C8 esters of β-hydroxybutyrate over a 4-hour period following injection are depicted.

[0017] Figure 7a-7h demonstrated that a ketogenic diet (but not fasting) consistently reduced epileptiform spikes in APPJ20 mice. Figure 7a ) A 23-hour EEG recorded 2 days after starting the KD showed a peak reduction of approximately 30% compared to the previous baseline on the control diet. An overnight fast showed no changes. ( Figure 7b ) Total number of peaks per hour during the 23-hour EEG recording. ( Figure 7c ) Peak reduction in individual mice, normalized to baseline recordings for each mouse (filled circles, P<0.05; bars = median). ( Figure 7d ) were similar in all dietary conditions, e.g. Figure 7eAs shown, overall normalized gamma activity. (Fig. 7f) Best fit linear regression line with 95% CI for scatter plots of peaks per minute and motility. APPJ20 generally had lower peaks with higher exploratory motility; with KD, peaks were lower at all motility levels. Figure 7g ) Best fit linear regression line with 95% CI for the scatter plot of normalized gamma activity and ambulation per minute, showing no change in the rate of gamma activity induction by exercise at KD. Figure 7h ) Overall mean gamma power was unchanged upon KD. P values ​​were compared by t-test for two-way comparisons and ANOVA with Tukey correction for multiple comparisons. N = 9-12 mice per condition; data for A, B, D, E show N = 7, which completed all conditions with high-quality data.

[0018] Figure 8a-8h Describe the KD-induced reduction in epileptiform spikes that persists for several months and is associated with cognitive improvements during open field adaptation. Figure 8a )Experimental timeline. Figure 8b ) In 7 50-minute EEGs, the KD had a peak reduction of APPJ20 of approximately 40% compared to mice on the control diet. Figure 8c ) Average peaks / minute during the 50-min recording period; peaks increased in mice on the control diet as exploratory activity faded later in the recording period. ( Figure 8d ) Best fit linear regression line with 95% CI for the scatter plot of peak per minute and exercise, showing that KD peaks are lower at all exercise levels. Figure 8e ) Total locomotion (beam breaks) during open field post-habituation at days 53 and 72 showed exploration was similar between APP and NTG controls at KD. Figure 8f -h) Total motion ( Figure 8f ),rear( Figure 8g ) and center motion ( Figure 8h ) were similar between APP and NTG controls at KD, indicating successful adaptation. P values ​​were by t-test for two-way comparisons and ANOVA with Tukey correction for multiple comparisons. N = 4-6 / group.

[0019] Figure 9a-9h Demonstrated that long-term ketogenic diet improves male cognition and survival. ( Figure 9a ) Body weight changes of APPJ20 and NTG mice on KD or control diet starting at 2 months of age. Figure 9b ) Plasma BHB levels were the average of six morning measurements taken approximately every two weeks from the start of the study. Fig.9c , 9d) Survival curves of APPJ20 males and females on KD and control diets, respectively. There was no mortality in NTG mice. ( Figure 9e-9h ) The Morris water maze was performed three months after the diet was started. The probe test was performed 24 hours after the last hidden platform training; the reversal training was started 24 hours after the initial probe test. APP mice on KD showed improved learning in both initial and reversal training ( Fig.9e , 9g), but no difference in the probe trial ( Figure 9f , 9h). P values ​​were obtained from t-test for two-way comparisons and ANOVA with Tukey correction for multiple comparisons. At the beginning of the study, N = 21-26 per genotype diet group; for water maze, N = 11-14 / group.

[0020] Figures 10a-10f Compounds described herein that are metabolized to BHB demonstrate an immediate reduction in epileptiform peaks. Fig.10a ) Schematic representation of an example of a ketogenic compound having a medium chain fatty acid ester linked to BHB. Figure 10b-10f ) Groups of mice were injected with C6-BHB and saline on different days, and EEG was recorded before and after each injection. Fig.10b ) Injection of C6-BHB increased blood BHB levels measured approximately 70 to 80 minutes after injection (post-EEG). Fig.10c ) Injection of C6-BHB reduced the peak compared to pre-injection baseline and saline injection. ( Fig.10d ) The curve of the mean peak of the 50-min EEG recording showed a consistent reduction after C6-BHB injection, which was similar to KD. Fig.10e ) Analysis of peak reduction after C6-BHB showed significant reduction at the individual mouse level for most mice compared to after saline injection (closed circles, P<0.05; bars = median). Fig.10f ) The peak difference between C6-BHB and saline injections was most apparent when mice were resting (and gamma activity was lowest), similar to KD. P values ​​were obtained from t-tests for two-way comparisons and ANOVA with Tukey correction for multiple comparisons. N = 22, and analysis was limited to 17 mice that completed all conditions with high-quality data.

[0021] Fig.11 The intake of food containing different concentrations of C6 ester of β-hydroxybutyrate over time is depicted.

[0022] Fig.12 Depicted is weight loss over time in mice following ingestion of a diet containing varying amounts of the C6 ester of β-hydroxybutyrate.

[0023] Fig.13 Depicted are blood glucose levels over time in mice when they were fed a diet containing varying amounts of the C6 ester of β-hydroxybutyrate.

[0024] Fig.14 Depicted are blood concentrations of β-hydroxybutyrate over time after mice were fed a diet with varying amounts of a C6 ester of β-hydroxybutyrate.

[0025] Fig.15 The intake of foods containing different esters of butanediol and β-hydroxybutyrate is depicted over time.

[0026] Fig.16 Depicted are weight changes over time in mice when fed a food composition supplemented with butanediol and different esters of β-hydroxybutyrate.

[0027] Fig.17 Depicted is the change in blood glucose levels over time in mice when fed a diet supplemented with butanediol and different esters of β-hydroxybutyrate.

[0028] Fig.18 Depicted is the change in blood concentration of β-hydroxybutyrate over time after mice were fed a diet containing butanediol and different esters of β-hydroxybutyrate.

[0029] Fig.19 Depicted are comparisons of plasma β-hydroxybutyrate, blood glucose, and caloric intake in mice consuming a control diet and mice consuming a diet supplemented with a C6 ester of β-hydroxybutyrate.

[0030] definition

[0031] As used herein, the terms "treat", "treating", and the like refer to obtaining a desired pharmacological and / or physiological effect. The effect may be preventive, in terms of completely or partially preventing a condition, disease, pathological process, or symptoms thereof, and / or may be therapeutic, in terms of partially or completely curing a condition, disease, pathological process, and / or side effects attributable to the condition, disease, or pathological process. As used herein, "treatment", for example, includes any treatment of a condition, disease, or pathological process in a mammal, particularly a human, and includes: (a) preventing the occurrence of a condition, disease, or pathological process in a subject who may be susceptible to the condition, disease, or pathological process but has not yet been diagnosed as having the condition, disease, or pathological process; (b) inhibiting a condition or disease, i.e., curbing its development; (c) alleviating a condition, disease, or pathological process, i.e., causing regression of the condition, disease, or pathological process.

[0032] The terms "individual," "subject," "host," and "patient," which are used interchangeably herein, refer to mammals, including but not limited to murines (rats, mice), non-human primates, humans, canines, felines, ungulates (e.g., horses, cattle, sheep, pigs, goats), and the like.

[0033] "Therapeutically effective amount" or "effective amount" refers to the amount of a compound that, when administered to a mammal or other subject for treating a disease, is sufficient to achieve said treatment for the disease. The "therapeutically effective amount" will vary depending on the compound or cell, the disease and its severity, and the age, weight, etc., of the subject to be treated.

[0034] The terms "co-administered" and "in combination with" include simultaneous, concurrent or sequential administration of two or more therapeutic agents without specific time limits. In one embodiment, the agents are present in a cell or subject at the same time or exert their biological or therapeutic effects at the same time. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, the first agent can be administered prior to (e.g., a few minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concurrently with, or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of the second therapeutic agent.

[0035] As used herein, "pharmaceutical composition" is intended to include compositions suitable for application to subjects (e.g., mammals, especially humans). Typically, "pharmaceutical composition" is sterile and does not contain contaminants that can cause undesirable reactions in the subject (e.g., the compound in the pharmaceutical composition is pharmaceutical grade). The pharmaceutical composition can be designed to be applied to subjects or patients in need by many different routes of administration, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, and intratracheal, etc. In some embodiments, the composition is suitable for administration by a transdermal route using a penetration enhancer other than dimethyl sulfoxide (DMSO). In other embodiments, the pharmaceutical composition is suitable for administration by a route other than transdermal administration. In some embodiments, the pharmaceutical composition includes a subject compound and a pharmaceutically acceptable excipient. In some embodiments, a pharmaceutically acceptable excipient is an excipient other than DMSO.

[0036] As used herein, "pharmaceutically acceptable derivatives" of the compounds of the invention include salts, esters, enol ethers, enol esters, acetals, ketals, orthoesters, hemiacetals, hemiketals, acids, bases, solvates, hydrates or prodrugs thereof. Those skilled in the art can readily prepare these derivatives using known methods for such derivatization. The resulting compounds can be administered to animals or humans without substantial toxic effects and are pharmaceutically active or prodrugs.

[0037] A "pharmaceutically acceptable salt" of a compound refers to a salt that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. These salts include: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or acid addition salts formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, or (2) salts formed when the acidic protons present in the parent compound are replaced by metal ions, such as alkali metal ions, alkaline earth metal ions or aluminum ions; or complexes with organic bases, such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

[0038] "Active agent" refers to a chemical substance or compound that exerts a pharmacological effect and is capable of treating, preventing or ameliorating one or more conditions / diseases as described herein (e.g., Alzheimer's disease). Examples of active agents of interest include fatty acid β-hydroxy ester compounds (e.g., fatty acid esters of β-hydroxybutyrate) and fatty acid esters of butanediol.

[0039] "Prodrug" refers to a derivative of an active agent that requires a transformation in vivo to release the active agent. In certain embodiments, the transformation is an enzymatic transformation. Prodrugs are often (although not necessarily) pharmacologically inactive until converted to an active agent.

[0040] Definition of selected chemical terms

[0041] The nomenclature of certain compounds or substituents is used in their conventional sense, such as described in the chemical literature, including but not limited to Loudon, Organic Chemistry, 4th edition, New York: Oxford University Press, 2002, pp. 360-361, 1084-1085; Smith and March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition, Wiley-Interscience, 2001.

[0042] As used herein, the term "alkanyl" by itself or as part of another substituent refers to a saturated branched or straight chain monovalent hydrocarbon group derived by removing a hydrogen atom from a single carbon atom of a parent alkane. Typical alkanyls include, but are not limited to, methyl; ethyl; propyl, such as prop-1-yl or prop-2-yl; and butyl, such as but-1-yl, but-2-yl, 2-methyl-prop-1-yl or 2-methyl-prop-2-yl. In certain embodiments, alkanyls contain 1-20 carbon atoms. In other embodiments, alkanyls contain 1 to 10 carbon atoms. In other embodiments, alkanyls contain 1 to 6 carbon atoms, such as 1 to 4 carbon atoms.

[0043] "Alkyl" by itself or as part of another substituent refers to a saturated branched, straight chain or cyclic alkyl group derived by removing a hydrogen atom from a single carbon atom of an alkane. Typical alkyl groups include, but are not limited to, methane; ethyl; propane, such as propan-1-yl, propan-2-yl (isopropyl), cyclopropane-1-yl, etc.; butane, such as butan-1-yl, butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl), 2-methyl-propan-2-yl (tert-butyl), cyclobutane-1-yl, etc.; and the like.

[0044] "Alkylene" refers to a branched or unbranched saturated hydrocarbon chain, typically having 1 to 40 carbon atoms, more typically 1 to 10 carbon atoms, and even more typically 1 to 6 carbon atoms. The term is exemplified by groups such as methylene (-CH2-), ethylene (-CH2CH2-), propylene isomers (e.g., -CH2CH2CH2- and -CH(CH3)CH2-), and the like.

[0045] "Alkenyl" by itself or as part of another substituent refers to an unsaturated branched, straight chain or cyclic hydrocarbon radical having at least one carbon-carbon double bond, derived by removing a hydrogen atom from a single carbon atom of an alkene. The radical may be in the cis or trans configuration about the double bond. Typical alkenyl groups include, but are not limited to, vinyl; propenyl groups such as prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyl groups such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, but-1,3-dien-1-yl, but-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobut-1,3-dien-1-yl, etc.; and the like.

[0046] "Alkynyl" by itself or as part of another substituent refers to an unsaturated branched, straight chain or cyclic hydrocarbon radical having at least one carbon-carbon triple bond, derived by removing a hydrogen atom from a single carbon atom of an alkyne. Typical alkynyl groups include, but are not limited to, ethynyl; propynyl, such as prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyl, such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.

[0047] "Acyl" by itself or as part of another substituent refers to the group -C(O)R 30 , where R as defined herein 30 is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, aralkyl, heteroalkyl, heteroaryl, heteroaralkyl, and substituted forms thereof. Representative examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl, piperonyl, succinyl, and malonyl, and the like.

[0048] The term "aminoacyl" refers to the group -C(O)NR 21 R 22 , where R 21 and R 22 are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclyl, and substituted heterocyclyl, and optionally, wherein R 21 and R 22are connected together with the nitrogen bonded thereto to form a heterocycle or a substituted heterocyclic group, and wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic group and substituted heterocyclic group are as defined herein.

[0049] "Alkoxy" by itself or as part of another substituent refers to the radical -OR 31 , where R 31 represents an alkyl or cycloalkyl group as defined herein. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclohexyloxy, and the like.

[0050] "Alkoxycarbonyl" by itself or as part of another substituent refers to the radical -C(O)OR 31 , where R 31 represents an alkyl or cycloalkyl group as defined herein. Representative examples include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, cyclohexyloxycarbonyl, and the like.

[0051] "Aryl" by itself or as part of another substituent refers to a monovalent aromatic hydrocarbon radical derived by removing one hydrogen atom from a single carbon atom of an aromatic ring system. Typical aryl groups include, but are not limited to, aryl groups derived from the following radicals: aceanthrene, acenaphthene, acetoxyphenanthrene, anthracene, azulene, benzene, In some embodiments, the aryl group comprises 6 to 20 carbon atoms. In certain embodiments, the aryl group contains 6 to 12 carbon atoms. Examples of aryl groups are phenyl and naphthyl.

[0052] "Aralkyl" by itself or as part of another substituent refers to an acyclic alkyl group in which the carbon atom (usually a terminal or sp 3 One of the hydrogen atoms bonded to a (C-carbon atom) is replaced by an aryl group. Typical aralkyl groups include, but are not limited to, benzyl, 2-phenylethyl-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethyl-1-yl, 2-naphthylethen-1-yl, naphthobenzyl, 2-naphthophenylethen-1-yl, etc. When it is intended to use a specific alkyl moiety, the aralkyl, arylalkenyl and / or arylalkynyl groups named are used. In certain embodiments, the aralkyl group is (C-C 30 ) aralkyl, for example, the alkyl, alkenyl or alkynyl portion of the aralkyl is (C1-C 10 ), the aryl part is (C6-C 20 In certain embodiments, the aralkyl group is (C7-C 20 ) aralkyl, for example, the alkyl, alkenyl or alkynyl portion of the aralkyl is (C1-C8) and the aryl portion is (C6-C 12 ).

[0053] "Arylaryl" by itself or as part of another substituent refers to a monovalent hydrocarbon radical derived by removing a hydrogen atom from a single carbon atom of a ring system in which two or more of the same or different aromatic ring systems are directly linked together by a single bond, wherein the number of such direct ring links is one less than the number of aromatic ring systems involved. Typical arylaryl radicals include, but are not limited to, biphenyl, triphenyl, phenyl-naphthyl, binaphthyl, biphenyl-naphthyl, and the like. When specifying the number of carbon atoms in arylaryl, the number refers to the carbon atoms included in each aromatic ring. For example, (C5-C 14 )Arylaryl is an arylaryl group containing 5 to 14 carbon atoms in each aromatic ring, such as biphenyl, terphenyl, binaphthyl, phenylnaphthyl, etc. In certain embodiments, each aromatic ring system of the arylaryl group is independently (C5-C 14 In certain embodiments, each aromatic ring system of the arylaryl group is independently (C5-C 14 ) Aromatic. In certain embodiments, each aromatic ring system is the same, such as biphenyl, triphenyl, binaphthyl, trinaphthyl, etc.

[0054] "Cycloalkyl" by itself or as part of another substituent refers to a saturated or unsaturated cyclic alkyl group. Where a specific level of saturation is intended, the nomenclature "cycloalkyl" or "cycloalkenyl" is used. Typical cycloalkyl groups include, but are not limited to, groups derived from cyclopropane, cyclobutane, cyclopentane, cyclohexane, and the like. In certain embodiments, a cycloalkyl group is (C3-C 10 In certain embodiments, the cycloalkyl group is a (C3-C7)cycloalkyl group.

[0055] "Cycloheteroalkyl" or "heterocyclyl" by itself or as part of another substituent refers to a saturated or unsaturated cyclic alkyl radical in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced by the same or different heteroatoms. Typical heteroatoms used to replace carbon atoms include, but are not limited to, N, P, O, S, Si, and the like. When a specific level of saturation is intended, the term "cycloheteroalkyl" or "cycloheteroalkenyl" is used. Typical cycloheteroalkyl radicals include, but are not limited to, radicals derived from epoxides, aziridines, thiirane, imidazoline, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, quinuclidine, and the like.

[0056] "Heteroalkyl, heteroalkyl, heteroalkenyl and heteroalkynyl" by themselves or as part of another substituent refer to alkyl, alkyl, alkenyl and alkynyl groups, respectively, in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced by the same or different heteroatom groups. Typical heteroatom groups that may be included in these groups include, but are not limited to, -O-, -S-, -SS-, -OS-, -NR 37 R 38 -,.=NN=,-N=N-,-N=N-NR 39 R 40 , -PR 41 -,-P(O)2-, -POR 42 -, -OP(O)2-, -SO-, -S-(O)-, -SO2-, -SnR 43 R 44 - etc., where R 37 , R 38 , R 39 , R 40 , R 41 , R 42 , R 43 and R 44 and R is independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaralkyl or substituted heteroaralkyl.

[0057] "Heteroaryl" by itself or as part of another substituent refers to a monovalent heteroaromatic group derived by removing one hydrogen atom from a single atom of a heteroaromatic ring system. Typical heteroaryl groups include, but are not limited to, groups derived from acridine, arsindene, carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perylene, phenanthridine, o-phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyridine ring, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, benzodioxole, and the like. In certain embodiments, the heteroaryl group is a 5-20 membered heteroaryl group. In certain embodiments, the heteroaryl group is a 5-10 membered heteroaryl group. In certain embodiments, the heteroaryl group is a group derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole, oxazole and pyrazine.

[0058] "Heteroaralkyl" by itself or as part of another substituent refers to a group in which the carbon atoms (typically terminal or sp 3 In some embodiments, the heteroarylalkyl group is a 6-30 yuan heteroarylalkyl group, for example, the alkyl, alkenyl or alkynyl moiety of the heteroarylalkyl group is 1-10 yuan, and the heteroaryl moiety is a 5-20 yuan heteroaryl group. In some embodiments, the heteroarylalkyl group is a 6-20 yuan heteroarylalkyl group, for example, the alkyl, alkenyl or alkynyl moiety of the heteroarylalkyl group is 1-8 yuan, and the heteroaryl moiety is a 5-12 yuan heteroaryl group.

[0059] "Aromatic ring system" by itself or as part of another substituent refers to an unsaturated cyclic or polycyclic ring system with a conjugated π electron system. The definition of "aromatic ring system" specifically includes fused ring systems in which one or more rings are aromatic and one or more rings are saturated or unsaturated, such as fluorene, indane, indene, phenalene, etc. Typical aromatic ring systems include, but are not limited to, acenaphthene, acenaphthene, phenanthracene, anthracene, azulene, benzene, Coronene, fluoranthene, fluorene, hexacene, hexylene, hexalene, asymmetric indacene, symmetric indacene, indane, indene, naphthyl, octacene, cinphene, aldrin, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, perylene, benzonaphthalene, phenanthrene, phenanthrene, septalene, pyrene, pyranthene, rubrocene, triphenylene, ternaphthalene, etc.

[0060] "Heteroaromatic ring system" by itself or as part of another substituent refers to an aromatic ring system in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced with the same or different heteroatoms. Typical heteroatoms used to replace carbon atoms include, but are not limited to, N, P, O, S, Si, and the like. Specifically included within the definition of "heteroaromatic ring system" are fused ring systems in which one or more rings are aromatic and one or more rings are saturated or unsaturated, for example, arsindenes, benzodioxanes, benzofurans, chromanes, chromenes, indoles, indolines, xanthenes, and the like. Typical heteroaromatic ring systems include, but are not limited to, arsindene, carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole, indole, dihydroindole, indolizine, isobenzofuran, isochromene, isoindole, isodihydroindole, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perylene, phenanthridine, o-phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyridine ring, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and the like.

[0061] "Substituted" refers to a group in which one or more hydrogen atoms are independently replaced by the same or different substituents. Typical substituents include, but are not limited to, alkylenedioxy (such as methylenedioxy), -M, -R 60 , -O - , =O, -OR 60 、-SR 60 , -S - , =S, -NR 60 R 61 , =NR 60 , -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2O - 、-S(O)2OH、-S(O)2R 60 、-OS(O)2O - 、-OS(O)2R 60 、-P(O)(O - )2、-P (O)(OR 60 )(O - )、-OP(O)(OR 60 )(OR 61 )、-C(O)R 60 、-C(S)R 60 、-C(O)OR 60 、-C(O)NR 60 R 61 、 -C(O)O - 、-C(S)OR 60 、-NR 62C(O)NR 60 R 61 、-NR 62 C(S)NR 60 R 61 、-NR 62 C(NR 63 )NR 60 R 61 and -C(NR 62 )NR 60 R 61 , wherein M is halogen; R 60 , R 61 , R 62 and R 63 are independently hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl, or alternatively, R 60 and R 61 Together with the nitrogen atom to which they are bound, they form a cycloheteroalkyl group or a cycloheteroalkyl group having a substituent; and R 64 and R 65 are independently hydrogen, alkyl, substituted alkyl, aryl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl, or alternatively, R 64 and R 65 Together with the nitrogen atom to which they are bound, they form a cycloheteroalkyl group or a cycloheteroalkyl ring having a substituent. In certain embodiments, the substituents include -M, -R 60 ,=O,-OR 60 、-SR 60 , -S - , =S, -NR 60 R 61 , =NR 60 , -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2R 60 、-OS(O)2O - 、-OS(O)2R 60 、-P(O)(O-)2、-P(O)(OR 60 )(O - )、-OP( O)(OR 60 )(OR 61 )、-C(O)R 60 、-C(S)R 60 、-C(O)OR 60 、-C(O)NR60 R 61 、-C(O)O - 、-NR 62 C(O)NR 60 R 61 In certain embodiments, the substituents include -M, -R 60 , =O, -OR 60 、-SR 60 、-NR 60 R 61 , -CF3, -CN, -NO2, -S(O)2R 60 、-P(O)(OR 60 )(O - )、-OP (O)(OR 60 )(OR 61 )、-C(O)R 60 、-C(O)OR 60 、-C(O)NR 60 R 61 、-C(O)O - In certain embodiments, the substituents include -M, -R 60 , =O, -OR 60 、-SR 60 、-NR 60 R 61 , -CF3, -CN, -NO2, -S(O)2R 60 、-OP(O)(OR 60 )(OR 61 )、- C(O)R 60 、-C(O)OR 60 、-C(O)O - , where R 60 , R 61 and R 62 For example, a substituted group may carry a methylenedioxy substituent or one, two or three substituents selected from halogen atoms, (1-4C)alkyl and (1-4C)alkoxy.

[0062] The compounds described herein may contain one or more chiral centers and / or double bonds and may therefore exist as stereoisomers, such as double bond isomers (i.e., geometric isomers), enantiomers, or diastereomers. Therefore, all possible enantiomers and stereoisomers of compounds including pure stereoisomeric forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers are included in the description of the compounds herein. Enantiomers and mixtures of stereoisomers can be separated into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to those skilled in the art. The compounds may also exist in several tautomeric forms, including enol forms, keto forms, and mixtures thereof. Therefore, the chemical structures described herein include all possible tautomeric forms of the compounds shown. The compounds also include isotopically labeled compounds in which the atomic mass of one or more atoms is different from the atomic mass conventionally found in nature. Examples of isotopes that can be incorporated into the compounds disclosed herein include, but are not limited to 2 H. 3 H. 11 C. 13 C. 14 C. 15 N. 18 O. 17 O, etc. The compounds may exist in unsolvated forms as well as solvated forms, including hydrated forms. Generally, the compounds may be hydrated or solvated. Certain compounds may exist in a variety of crystalline or amorphous forms. Generally, all physical forms are equivalent for the uses contemplated herein and are intended to fall within the scope of the present disclosure.

[0063] Before further describing the present invention, it should be understood that the present invention is not limited to the particular embodiments described, which may, of course, vary. It should also be understood that the terminology used herein is only for the purpose of describing particular embodiments and is not restrictive, as the scope of the present invention is limited only by the appended claims.

[0064] Where a range of values ​​is provided, it is to be understood that, unless the context clearly dictates otherwise, each intervening value between the upper and lower limits of the range, to one-tenth of the unit of the lower limit, and any other stated or intermediate value within the stated range, is included within the present invention. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges, and are also included within the present invention, subject to any explicitly excluded limitations within the stated ranges. Where the stated range includes one or two limitations, ranges excluding one or both of those included limitations are also included within the present invention.

[0065] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials associated with the cited publications.

[0066] It must be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, this statement is intended to serve as antecedent basis for use of the terminology "solely," "only," and the like in connection with the recitation of claim elements, or use of a "negative" limitation.

[0067] It should be understood that certain features of the invention described in the context of separate embodiments for the sake of clarity may also be provided in combination in a single embodiment. Conversely, various features of the invention described in the context of a single embodiment for the sake of brevity may also be provided individually or in any suitable subcombination. The present invention specifically includes all combinations of embodiments related to the present invention and are disclosed herein as if each and every combination were individually and explicitly disclosed. In addition, all subcombinations of various embodiments and elements thereof are also specifically included in the present invention and are disclosed herein as if each and every such subcombination were individually and explicitly disclosed.

[0068] The publications discussed herein are provided only for their disclosures prior to the filing date of the present application. Anything herein should not be construed as admitting that the present invention is not entitled to precede such publications. In addition, the publication date provided may be different from the actual publication date and may need to be confirmed separately. DETAILED DESCRIPTION

[0069] The present disclosure provides fatty acid β-hydroxy ester compounds (e.g., fatty acid esters of β-hydroxybutyrate), fatty acid esters of butanediol, and pharmaceutically acceptable salts thereof. Also provided are pharmaceutical compositions having one or more fatty acid β-hydroxy ester compounds and / or one or more fatty acid esters of butanediol.

[0070] The present disclosure also provides a method for treating a subject by administering one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters to the subject. In certain embodiments, the method includes treating Alzheimer's disease in a subject diagnosed with Alzheimer's disease by administering one or more fatty acid β-hydroxy ester compounds as described herein and / or one or more butylene glycol esters as described herein to the subject. In some cases, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the subject is sufficient to reduce the epileptic activity of the subject. In some cases, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the subject is sufficient to increase the cognition of the subject.

[0071] The present disclosure further provides methods of treating one or more of the following diseases: epilepsy, Parkinson's disease, heart failure, traumatic brain injury, stroke, hemorrhagic shock, acute lung injury after fluid resuscitation, acute kidney injury, myocardial infarction, myocardial ischemia, diabetes, glioblastoma multiforme, diabetic neuropathy, prostate cancer, amyotrophic lateral sclerosis, Huntington's disease, cutaneous T-cell lymphoma, multiple myeloma, peripheral T-cell lymphoma, HIV, Niemann-Pick type C disease, age-related macular degeneration, gout, atherosclerosis, rheumatoid arthritis and multiple sclerosis by administering to a subject one or more fatty acid β-hydroxy ester compounds described herein and / or one or more butanediol esters described herein.

[0072] The present disclosure also provides compounds, and the use of the compounds provides an increase in ketone body concentration in a subject. As described herein, fatty acid β-hydroxy ester compounds (e.g., fatty acid esters of β-hydroxybutyrate) and fatty acid esters of butanediol release β-hydroxybutyrate or butanediol by ester hydrolysis, which increases the ketone body concentration in the subject and the supplementary source of medium-chain fatty acids, and medium-chain fatty acids provide ketone bodies that continue to be produced in the subject. In certain embodiments, the present disclosure provides fatty acid esters of β-hydroxybutyrate, which, after administration, are hydrolyzed into β-hydroxybutyrate and medium-chain fatty acids. In these embodiments, the release of β-hydroxybutyrate provides an increase in ketone body concentration in the subject, and the medium-chain fatty acids of hydrolysis provide substrates for the continued physiological production of ketone bodies. In other embodiments, the present disclosure provides fatty acid esters of butanediol, which, after administration, are hydrolyzed into butanediol and medium-chain fatty acids. In these embodiments, The release of butanediol provides an increase in ketone body production in the subject, and the medium-chain fatty acids of hydrolysis provide additional substrates for the continued physiological production of ketone bodies.

[0073] Fatty acid β-hydroxy ester compounds and fatty acid esters of butanediol

[0074] Fatty acid β-hydroxy ester compounds and fatty acid esters of butanediol (described in more detail below) suitable for use in practicing the process of the present invention include compounds of Formulas I and II as described below.

[0075] Compounds of formula I and II

[0076] The compositions of the present invention include compounds of Formula I and II as shown below. The pharmaceutical compositions and methods of the present disclosure also encompass compounds of Formula I and II.

[0077] In one aspect of its composition, embodiments of the present invention provide a compound of formula I:

[0078] in

[0079] R1 is selected from the group consisting of hydrogen, alkyl, and alkyl having a substituent; and

[0080] R2 and R3 are independently an alkyl group which has no substituent or has a substituent;

[0081] and its salts, solvates or hydrates.

[0082] In Formula I, R1 is selected from hydrogen, alkyl and alkyl with a substituent. In some cases, R1 is hydrogen. In other cases, R1 is alkyl. In other cases, R1 is alkyl with a substituent. In some cases, R1 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R1 is methyl.

[0083] In Formula I, R2 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R2 is an alkyl group. In other cases, R2 is an alkyl group having a substituent. In some cases, R2 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R2 is hexyl or octyl (C6 or C8). In some cases, R2 is hexyl (C6). In some cases, R2 is octyl (C8).

[0084] In Formula I, R3 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R3 is an alkyl group. In other cases, R3 is an alkyl group having a substituent. In some cases, R3 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R3 is hexyl or octyl (C6 or C8). In some cases, R3 is hexyl (C6). In some cases, R3 is octyl (C8).

[0085] In Formula I according to certain embodiments, R1 is methyl and R2 and R3 are independently hexyl. In some cases, R1 is methyl and R2 and R3 are independently octyl. In other cases, R1 is methyl and R2 is hexyl and R3 is octyl. In other cases, R1 is methyl and R2 is octyl and R3 is hexyl.

[0086] In certain embodiments, the fatty acid ester of 1,3-butanediol of interest is enantiomerically pure (<95% ee). 1,3-Butanediol is metabolized to BHB by alcohol dehydrogenase and aldehyde dehydrogenase, both of which act on the 1' hydroxyl group and have no specificity for the chirality of the 3' group. Thus, R-1,3-butanediol is metabolized to R-BHB by these enzymes, and S-1,3-butanediol is metabolized to S-BHB by these enzymes.

[0087] The compounds described in this application include 1,3-butanediol esters linked to one or more fatty acids or BHB esters linked to one or more fatty acids. The compound is chiral because the 1,3-butanediol and BHB moieties are chiral. However, it is important to note that due to the stereospecificity of the above-mentioned β-hydroxybutyrate dehydrogenase, the fatty acid moiety can only be metabolized to R-BHB in vivo. Therefore, a compound containing S-BHB linked to a fatty acid will ultimately produce one unit of S-BHB, but also produce several units of R-BHB. In certain embodiments, when completely metabolized in vivo, none of the compounds described herein produce S-BHB.

[0088] In certain embodiments, the metabolic differences between R-BHB and S-BHB are important for their relative efficacy in treating human diseases. As mentioned above, only R-BHB can be easily metabolized into acetyl-CoA and ATP, thereby realizing the "energy currency" function of BHB. If the efficacy of treating diseases is based on this energy function of BHB, then R-BHB will be substantially more effective than S-BHB. However, in some cases, the efficacy of BHB in certain human diseases may rely on the signal transduction function of BHB, rather than the energy function. Several signal transduction functions of concern may include but are not limited to HCAR2 inhibition, inflammasome inhibition and inhibition of deacetylase. In these embodiments, wherein S-BHB has a molecular effect similar to R-BHB, then the slower metabolism of S-BHB can provide a longer duration of action in vivo.

[0089] In another composition aspect, an embodiment of the present invention provides a compound of Formula Ia:

[0090] in

[0091] R1 is selected from the group consisting of hydrogen, alkyl, and alkyl having a substituent; and

[0092] R2 and R3 are independently an alkyl group which has no substituent or has a substituent;

[0093] and its salts, solvates or hydrates.

[0094] In Formula Ia, R1 is selected from hydrogen, alkyl and substituted alkyl. In some cases, R1 is hydrogen. In other cases, R1 is alkyl. In other cases, R1 is substituted alkyl. In some cases, R1 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R1 is methyl.

[0095] In Formula Ia, R2 is selected from alkyl and substituted alkyl. In some cases, R2 is alkyl. In other cases, R2 is substituted alkyl. In some cases, R2 is alkyl, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R2 is hexyl or octyl (C6 or C8). In some cases, R2 is hexyl (C6). In some cases, R2 is octyl (C8).

[0096] In Formula Ia, R3 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R3 is an alkyl group. In other cases, R3 is an alkyl group having a substituent. In some cases, R3 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R3 is hexyl or octyl (C6 or C8). In some cases, R3 is hexyl (C6). In some cases, R3 is octyl (C8).

[0097] In Formula Ia according to certain embodiments, R1 is methyl and R2 and R3 are independently hexyl. In some cases, R1 is methyl and R2 and R3 are independently octyl. In other cases, R1 is methyl and R2 is hexyl and R3 is octyl. In other cases, R1 is methyl and R2 is octyl and R3 is hexyl.

[0098] In certain embodiments, fatty acid esters of butanediol of interest include compounds of formula BDE-1a to BDE-1b:

[0099] and

[0100]

[0101] and its salts, solvates or hydrates.

[0102] In another composition aspect, an embodiment of the present invention provides a compound of Formula Ib:

[0103] in

[0104] R1 is selected from the group consisting of hydrogen, alkyl, and alkyl having a substituent; and

[0105] R2 and R3 are independently an alkyl group which has no substituent or has a substituent.

[0106] and its salts, solvates or hydrates.

[0107] In Formula Ib, R1 is selected from hydrogen, alkyl and substituted alkyl. In some cases, R1 is hydrogen. In other cases, R1 is alkyl. In other cases, R1 is substituted alkyl. In some cases, R1 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R1 is methyl.

[0108] In Formula Ib, R2 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R2 is an alkyl group. In other cases, R2 is an alkyl group having a substituent. In some cases, R2 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R2 is hexyl or octyl (C6 or C8). In some cases, R2 is hexyl (C6). In some cases, R2 is octyl (C8).

[0109] In Formula Ib, R3 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R3 is an alkyl group. In other cases, R3 is an alkyl group having a substituent. In some cases, R3 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R3 is hexyl or octyl (C6 or C8). In some cases, R3 is hexyl (C6). In some cases, R3 is octyl (C8).

[0110] In formula Ib according to certain embodiments, R1 is methyl and R2 and R3 are independently hexyl. In some cases, R1 is methyl and R2 and R3 are independently octyl. In other cases, R1 is methyl and R2 is hexyl and R3 is octyl. In other cases, R1 is methyl and R2 is octyl and R3 is hexyl.

[0111] In another composition aspect, an embodiment of the present invention provides a compound of formula II:

[0112] in

[0113] R4 is selected from the group consisting of hydrogen, alkyl, and substituted alkyl; and

[0114] R5 and R6 are independently an alkyl group which has no substituent or has a substituent;

[0115] and its salts, solvates or hydrates.

[0116] In formula II, R4 is selected from hydrogen, alkyl and alkyl with substituents. In some cases, R4 is hydrogen. In other cases, R4 is alkyl. In other cases, R4 is alkyl with substituents. In some cases, R4 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R4 is methyl.

[0117] In Formula II, R5 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R5 is an alkyl group. In other cases, R5 is an alkyl group having a substituent. In some cases, R5 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R5 is hexyl or octyl (C6 or C8). In some cases, R5 is hexyl (C6). In some cases, R5 is octyl (C8).

[0118] In Formula II, R6 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R6 is an alkyl group. In other cases, R6 is an alkyl group having a substituent. In some cases, R6 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R 3 In some cases, R6 is hexyl (C6). In some cases, R 3 It is octyl (C8).

[0119] In Formula II according to certain embodiments, R4 is methyl and R5 and R6 are independently hexyl. In some cases, R4 is methyl and R5 and R6 are independently octyl. In other cases, R4 is methyl and R5 is hexyl and R6 is octyl. In other cases, R4 is methyl and R5 is octyl and R6 is hexyl.

[0120] In certain embodiments, the fatty acid β-hydroxy ester compound of interest is enantiomerically pure (<95% ee). R-BHB is a normal product of human metabolism. This chiral specificity is introduced by an enzyme (β-hydroxybutyrate dehydrogenase) that catalyzes the final step of BHB synthesis. The enzyme reduces the 3' carbonyl of acetoacetate to the 3' hydroxyl of BHB. The same enzyme is also required to utilize BHB by catalyzing the same reaction in reverse. Due to the chiral specificity of β-hydroxybutyrate dehydrogenase, only R-BHB is produced by normal metabolism, and only R-BHB can be easily broken down into acetyl-CoA and ATP. Fasting, exercise, caloric restriction, ketogenic diet, and any other state that leads to endogenous production of BHB will only produce R-BHB.

[0121] S-BHB itself is not a normal product of human metabolism. However, S-BHB-CoA is a transient intermediate in the final round of beta-oxidation of fatty acids. Under normal circumstances, it should not last long enough to leave the mitochondria or circulate in the blood. Experiments involving infusions of labeled R-BHB, S-BHB, or mixtures into rats or pigs found that S-BHB was converted primarily to R-BHB; the molecular pathway for this is unknown, but may be through conversion of S-BHB to acetyl-CoA, from which R-BHB is then produced. At least some of the S-BHB is ultimately converted to CO2, presumably after being metabolized to acetyl-CoA, which is then metabolized in the TCA cycle. S-BHB is likely metabolized much more slowly than R-BHB, so an infusion of the same amount of S-BHB may result in elevated blood levels of S-BHB for a longer period of time than a similar infusion of R-BHB would produce levels of R-BHB in the blood.

[0122] In another composition aspect, an embodiment of the present invention provides a compound of Formula IIa:

[0123] in

[0124] R4 is selected from the group consisting of hydrogen, alkyl, and substituted alkyl; and

[0125] R5 and R6 are independently an alkyl group which has no substituent or has a substituent;

[0126] and its salts, solvates or hydrates.

[0127] In formula IIa, R4 is selected from hydrogen, alkyl and alkyl with substituents. In some cases, R4 is hydrogen. In other cases, R4 is alkyl. In other cases, R4 is alkyl with substituents. In some cases, R4 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R4 is methyl.

[0128] In Formula IIa, R5 is selected from alkyl and substituted alkyl. In some cases, R5 is alkyl. In other cases, R5 is substituted alkyl. In some cases, R5 is alkyl, such as C 4- C 30 Alkyl, including C6-C8 alkyl. In some cases, R5 is hexyl or octyl (C6 or C8). In some cases, R5 is hexyl (C6). In some cases, R5 is octyl (C8).

[0129] In Formula IIa, R6 is selected from an alkyl group and an alkyl group having a substituent. In some cases, R6 is an alkyl group. In other cases, R6 is an alkyl group having a substituent. In some cases, R6 is an alkyl group, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R 3 In some cases, R6 is hexyl (C6). In some cases, R 3 It is octyl (C8).

[0130] In formula IIa according to certain embodiments, R4 is methyl and R5 and R6 are independently hexyl. In some cases, R4 is methyl and R5 and R6 are independently octyl. In other cases, R4 is methyl and R5 is hexyl and R6 is octyl. In other cases, R4 is methyl and R5 is octyl and R6 is hexyl.

[0131] In certain embodiments, fatty acid β-hydroxy ester compounds of interest include β-hydroxybutyrate compounds of formula BHE-2a to BHE-2d:

[0132]

[0133] and

[0134]

[0135] and its salts, solvates or hydrates.

[0136] In another composition aspect, an embodiment of the present invention provides a compound of Formula IIa:

[0137] in

[0138] R4 is selected from the group consisting of hydrogen, alkyl, and substituted alkyl; and

[0139] R5 and R6 are independently an alkyl group which has no substituent or has a substituent;

[0140] and its salts, solvates or hydrates.

[0141] In formula IIb, R4 is selected from hydrogen, alkyl and alkyl with substituents. In some cases, R4 is hydrogen. In other cases, R4 is alkyl. In other cases, R4 is alkyl with substituents. In some cases, R4 is alkyl, such as C1-C6 alkyl, including C1-C3 alkyl. In some cases, R4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl. In some cases, R4 is methyl.

[0142] In Formula IIb, R5 is selected from alkyl and substituted alkyl. In some cases, R5 is alkyl. In other cases, R5 is substituted alkyl. In some cases, R5 is alkyl, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R5 is hexyl or octyl (C6 or C8). In some cases, R5 is hexyl (C6). In some cases, R5 is octyl (C8).

[0143] In Formula IIb, R6 is selected from alkyl and substituted alkyl. In some cases, R6 is alkyl. In other cases, R6 is substituted alkyl. In some cases, R6 is alkyl, such as C4-C 30 Alkyl, including C6-C8 alkyl. In some cases, R 3 In some cases, R6 is hexyl (C6). In some cases, R 3 It is octyl (C8).

[0144] In formula IIb according to certain embodiments, R4 is methyl and R5 and R6 are independently hexyl. In some cases, R4 is methyl and R5 and R6 are independently octyl. In other cases, R4 is methyl and R5 is hexyl and R6 is octyl. In other cases, R4 is methyl and R5 is octyl and R6 is hexyl.

[0145] Formulation, dosage and route of administration

[0146] Pharmaceutically acceptable carriers preferably used together with the active agent (and optionally, one or more additional therapeutic agents) may include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcohol / water solutions, emulsions or suspensions and microparticles, including saline and buffered media. Parenteral carriers include sodium chloride solution, Ringer's dextrose, glucose and sodium chloride, lactated Ringer's solution or fixed oils. Intravenous carriers include liquid and nutritional supplements, electrolyte supplements (e.g., supplements based on Ringer's dextrose), etc. Compositions comprising the active agent (and optionally, one or more additional therapeutic agents) may also be freeze-dried using methods well known in the art for subsequent reconstruction and use according to the present invention.

[0147] preparation

[0148] The fatty acid ester of the subject fatty acid β-hydroxy ester compound and / or the subject butanediol can be administered to individuals in need in a preparation with a pharmaceutically acceptable excipient. A variety of pharmaceutically acceptable excipients are known in the art and do not need to be discussed in detail herein. Pharmaceutically acceptable excipients are fully described in various publications, For example, they include A.Gennaro (2000) "Remington: The Science and Practice of Pharmacy", 20th edition, Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) HC Ansel et al., 7th edition, Lippincott, Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) AK Kibbe et al., 3rd edition. Amer. Pharmaceutical Assoc. For purposes of the following formulation description, "active agent" includes active agents as described above, for example, fatty acid β-hydroxy ester compounds or fatty acid esters of butanediol as described herein, and optionally, one or more additional therapeutic agents.

[0149] In the subject method, any convenient scheme can be used to apply the active agent to the host. Therefore, the active agent can be incorporated into various preparations for therapeutic administration. For example, the active agent can be formulated into a pharmaceutical composition by combining with a suitable pharmaceutically acceptable carrier or diluent, and can be formulated into a preparation in solid, semisolid, liquid or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols. In an exemplary embodiment, the active agent is formulated into a gel, solution or some other form suitable for intravaginal administration. In another exemplary embodiment, the active agent is formulated into a gel, solution or some other form suitable for rectal (e.g., intrarectal) administration.

[0150] In pharmaceutical dosage forms, the active agent may be administered in the form of its pharmaceutically acceptable salt, or it may be used alone or in appropriate combination, as well as in combination with other pharmaceutically active compounds. The following methods and excipients are merely exemplary and are by no means limiting.

[0151] In certain embodiments, the active is formulated in an aqueous buffer. Suitable aqueous buffers include, but are not limited to, acetate, succinate, citrate, and phosphate buffers, with strengths varying between about 5 mM and about 100 mM. In certain embodiments, the aqueous buffer includes an agent that provides an isotonic solution. These agents include, but are not limited to, sodium chloride; and sugars, such as mannitol, glucose, sucrose, and the like. In certain embodiments, the aqueous buffer further includes a nonionic surfactant, such as polysorbate 20 or 80. Optionally, the formulation may further include a preservative. Suitable preservatives include, but are not limited to, benzyl alcohol, phenol, chlorobutanol, benzalkonium chloride, and the like. In many cases, the formulation is stored at about 4°C. The formulation may also be lyophilized, in which case they typically include cryoprotectants, such as sucrose, trehalose, lactose, maltose, mannitol, and the like. Lyophilized formulations can be stored for a long time, even at ambient temperature.

[0152] For oral preparations, the active agent can be used alone or in combination with appropriate additives to prepare tablets, powders, granules or capsules, for example, in combination with conventional additives such as lactose, mannitol, corn starch or potato starch; in combination with binders such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatin; in combination with disintegrants such as corn starch, potato starch or sodium carboxymethyl cellulose; in combination with lubricants such as talc or magnesium stearate; and if necessary, in combination with diluents, buffers, wetting agents, preservatives and flavoring agents.

[0153] The active agents described herein can be provided and / or administered as a food supplement, for example, in combination with one or more components of a ketogenic diet. Exemplary ketogenic diets and their components are described, for example, in U.S. Pat. No. 6,207,856, the disclosure of which is incorporated herein by reference.

[0154] The active agent can be formulated into an injectable preparation by dissolving, suspending or emulsifying the active agent in an aqueous or non-aqueous solvent, such as vegetable oil or other similar oils, synthetic fatty acid glycerides, higher fatty acid esters or propylene glycol; and if necessary, conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives can be used.

[0155] The active agent can be used in an aerosol formulation for administration by inhalation. The active agent can be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0156] In addition, the active agent can be made into a suppository by mixing with various bases such as an emulsifying base or a water-soluble base. The active agent can be administered rectally via a suppository. Suppositories can include carriers such as cocoa butter, carbowax, and polyethylene glycol, which melt at body temperature but solidify at room temperature.

[0157] Unit dosage forms for oral or rectal administration, such as syrups, elixirs and suspensions, can be provided, wherein each dosage unit (e.g., a teaspoon, tablespoon, tablet or suppository) contains a predetermined amount of a composition containing one or more active agents. Similarly, unit dosage forms for injection or intravenous administration can contain the active agent in the composition as a solution in sterile water, saline or another pharmaceutically acceptable carrier.

[0158] Unit dosage forms for intravaginal or intrarectal administration, such as syrups, elixirs, gels and suspensions, can be provided, wherein each dosage unit (e.g., a teaspoon, a tablespoon, a tablet, a unit gel volume or a suppository) contains a predetermined amount of the composition containing one or more active agents.

[0159] As used herein, the term "unit dosage form" refers to physically discrete units suitable as unit dosages for human and animal subjects, each unit containing a predetermined quantity of active agent calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for a given active agent will depend in part on the specific compound employed and the effect to be achieved, as well as the pharmacodynamics associated with each compound in the host.

[0160] Other modes of administration may also be used with the present invention. For example, the active agent may be formulated as a suppository, and in some cases, as an aerosol and intranasal composition. For a suppository, the carrier composition will include a conventional binder and a carrier, such as a polyalkylene glycol or a triglyceride. Such a suppository may be formed from a mixture containing an active ingredient in an amount ranging from about 0.5% to about 25% (w / w), such as from about 5% to about 20%, including 5% to 15%.

[0161] The active agent can be administered as an injection. Typically, injectable compositions are prepared as liquid solutions or suspensions; solid forms suitable for dissolution or suspension in a liquid carrier prior to injection can also be prepared. The formulation can also be emulsified or the active ingredient can be encapsulated in a liposome carrier.

[0162] In certain embodiments, the active agent will be formulated for vaginal delivery. The subject formulations for intravaginal administration comprise the active agent formulated as an intravaginal bioadhesive tablet, intravaginal bioadhesive microparticles, intravaginal cream, intravaginal wash, intravaginal foam, intravaginal ointment, intravaginal paste, intravaginal solution, or intravaginal gel.

[0163] In certain embodiments, the active agent will be formulated for rectal delivery. The subject formulations for intrarectal administration comprise the active agent formulated as an intrarectal bioadhesive tablet, intrarectal bioadhesive microparticles, intrarectal cream, intrarectal wash, intrarectal foam, intrarectal ointment, intrarectal paste, intrarectal solution, or intrarectal gel.

[0164] The subject formulations comprising the active agent include one or more of the following: excipients (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate or calcium carbonate), binders (e.g., cellulose, methylcellulose, hydroxymethylcellulose, polypropyl pyrrolidone, polyvinyl pyrrolidone, gelatin, gum arabic, poly(ethylene glycol), sucrose or starch), disintegrants (e.g., starch, carboxymethylcellulose, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, sodium bicarbonate, calcium phosphate or calcium citrate), lubricants (e.g., magnesium stearate, light anhydrous silicic acid, talc or sodium lauryl sulfate), flavoring agents (e.g., citric acid, menthol, glycine or orange powder), preservatives (e.g., sodium benzoate, sodium bisulfite, methylparaben or propylparaben), stabilizers (e.g., citric acid, sodium citrate or acetic acid), suspending agents (e.g., methylcellulose, Polyvinyl pyrrolidone or aluminum stearate), a dispersant (eg, hydroxypropyl methylcellulose), a diluent (eg, water) and a base wax (eg, cocoa butter, white petrolatum or polyethylene glycol).

[0165] Tablets containing the active agent may be coated with a suitable film-forming agent, such as hydroxypropylmethylcellulose, hydroxypropylcellulose or ethylcellulose, to which suitable excipients may optionally be added, such as a softener, such as glycerol, propylene glycol, diethyl phthalate or triacetin; a filler, such as sucrose, sorbitol, xylitol, glucose or lactose; a colorant, such as titanium hydroxide; and the like.

[0166] Suitable excipient carriers are, for example, water, saline, glucose, glycerol, ethanol, and the like, and combinations thereof. In addition, if desired, the carrier may contain a small amount of auxiliary substances, such as a wetting agent or emulsifier or a pH buffer. The actual method of preparing such dosage forms is known to those skilled in the art, or will be apparent. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 17th edition, 1985. The composition or formulation to be administered in any case contains an amount of the agent sufficient to achieve the desired state in the subject being treated.

[0167] Pharmaceutically acceptable excipients, such as carriers, adjuvants, vehicles or diluents, are readily available to the public. In addition, pharmaceutically acceptable auxiliary substances, such as pH adjusters and buffers, tonicity adjusters, stabilizers, wetting agents, etc., are readily available to the public.

[0168] dose

[0169] Although the dosage used will vary depending on the clinical goals to be achieved, suitable dosage ranges for the active agent are those that provide up to about 1 g to about 100 g, e.g., about 1 g to about 90 g, about 2.5 g to about 80 g, about 5.0 g to about 70 mg, about 7.5 g to about 60 g, about 10 g to about 50 g, about 12.5 g to about 40 g, about 15 g to about 30 g, or 5 g to 20 g of the active agent that can be administered in a single dose.

[0170] Those skilled in the art will readily appreciate that dosage levels may vary depending on the specific compound, the severity of the symptoms and the subject's susceptibility to side effects. Preferred dosages for a given compound can be readily determined by a variety of methods by those skilled in the art.

[0171] In certain embodiments, a single dose of the active agent is administered. In other embodiments, multiple doses of the active agent are administered. If multiple doses are administered over a period of time, the administration of the active agent is twice daily (qid), daily (qd), every other day (qod), every three days, three times / week (tiw) or twice / week (biw) administration over a period of time. For example, the active agent is administered qid, qd, qod, tiw or biw over a period of time from one day to about 2 years or more. For example, depending on various factors, the active agent is administered for one week, two weeks, one month, two months, six months, one year or more than two years at any of the above frequencies.

[0172] Thus, the amount of active agent administered to a subject daily may be 10-500 g / day, such as 15-450 g / day, such as 25-400 g / day, such as 50-300 g / day and including 100-200 g / day.

[0173] When two different active agents are administered, the first active agent and the second active agent can be administered in separate formulations. The first active agent and the second active agent can be administered substantially simultaneously, or within about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 8 hours, about 16 hours, about 24 hours, about 36 hours, about 72 hours, about 4 days, about 7 days, or about 2 weeks of each other.

[0174] It should be noted that a particular method, dosage, dosing regimen, route of administration, etc. described herein may not be suitable due to the form of a disclosed compound, e.g., due to the presence of a salt form of a disclosed compound. Thus, the form of the compound may be selected for use in a method, dosage, dosing regimen, route of administration, etc. For example, where the presence of a salt form of a disclosed compound is not suitable (e.g., due to the presence of a deleterious amount of salt in the dosage form), the form of the compound provided for a particular method, dosage, dosing regimen, route of administration, etc. may specifically exclude the salt form of the compound.

[0175] Route of administration

[0176] The active agent is administered to the individual using any available method and route suitable for drug delivery, including in vivo and ex vivo methods, and systemic and topical routes of administration.

[0177] Traditional and pharmaceutically acceptable routes of administration include oral, intranasal, intramuscular, intratracheal, transdermal, subcutaneous, intradermal, topical, intravenous, vaginal, nasal and other parenteral routes of administration. In certain embodiments, the activating agent is administered orally. In certain embodiments, the activating agent is administered via an intravaginal route of administration. In other embodiments, the activating agent is administered via an intrarectal route of administration. If desired, routes of administration may be combined, or routes of administration may be adjusted according to the agent and / or desired effect. The composition may be administered in a single dose or multiple doses.

[0178] The active agent may be administered to the host using any available conventional methods and routes suitable for the delivery of conventional drugs, including systemic or topical routes. In general, routes of administration contemplated by the present invention include, but are not necessarily limited to, enteral, parenteral or inhalation routes.

[0179] Parenteral administration routes other than administration by inhalation include, but are not necessarily limited to, topical, vaginal, transdermal, subcutaneous, intramuscular, and intravenous routes, i.e., any route of administration other than through the digestive tract. Parenteral administration may be performed to achieve systemic or local delivery of an agent. Where systemic delivery is desired, administration generally involves local or mucosal administration of invasive or systemic absorption of the pharmaceutical formulation.

[0180] The active agent may also be delivered to a subject by enteral administration. Enteral administration routes include, but are not necessarily limited to, oral and rectal (eg, using a suppository) delivery.

[0181] As used herein, the term "treatment" may refer to the improvement of symptoms associated with a pathological condition afflicting a host, wherein improvement is used in a broad sense to refer to a decrease in at least the magnitude of a parameter, e.g., a symptom associated with the pathological condition being treated, such as the presence of abnormal epileptiform spikes. Thus, treatment also includes situations where the pathological condition, or at least the symptoms associated therewith, are completely suppressed, e.g., prevented from occurring or stopped, e.g., terminated, such that the host no longer suffers from the pathological condition, or at least no longer suffers from the symptoms characterizing the pathological condition.

[0182] Compounds and compositions described herein can be applied to a variety of hosts (wherein the term "host" can be used interchangeably with the terms "subject" and "patient" herein). In certain embodiments, the subject is "mammal" or "mammalian", wherein these terms are widely used to describe organisms within the class of mammals, including carnivores (e.g., dogs and cats), rodents (e.g., mice, guinea pigs and rats) and primates (e.g., humans, chimpanzees and monkeys). In some cases, the subject is a human. Human subjects can be of any gender and at any stage of development (i.e., neonates, infants, teenagers, adolescents, adults), wherein in certain embodiments, human subjects are teenagers, adolescents or adults. Although the disclosure can be applied to human subjects, it should be understood that the fatty acid esters of the subject fatty acid β-hydroxy ester compounds and butanediol can also be applied to animal subjects (i.e., "non-human subjects"), such as, but not limited to, birds, mice, rats, dogs, cats, livestock and horses.

[0183] Kits, containers, devices, delivery systems

[0184] Kits are provided with unit doses of active agents, e.g., oral, vaginal, rectal, transdermal or injectable doses (e.g., for intramuscular, intravenous or subcutaneous injection). In such kits, in addition to the container containing the unit dose, there will also be an information package insert describing the use and attendant benefits of the subject composition. Suitable active agents and unit doses are as described above.

[0185] In many embodiments, the subject kit will also include instructions for practicing the subject method or a device for obtaining instructions for the subject method (e.g., directing a user to a website URL that provides an instruction web page), wherein these instructions are typically printed on a substrate, which can be one or more of the following: a package insert, a package, a formulation container, etc.

[0186] In certain embodiments, the subject kit includes one or more components or features that increase patient compliance, for example, a component or system that helps the patient remember to take the active agent at the appropriate time or interval. These components include, but are not limited to, a calendar system that helps the patient remember to take the active agent at the appropriate time or interval.

[0187] In certain embodiments, the delivery system is a delivery system that provides for subcutaneous, intravenous, or intramuscular injection of a formulation comprising an active agent. In other embodiments, the delivery system is a vaginal or rectal delivery system.

[0188] In certain embodiments, the active agent is packaged for oral administration. The present invention provides a packaging unit comprising an active agent in a daily dosage unit. For example, in certain embodiments, the packaging unit is a conventional blister package or any other form comprising tablets, pills, etc. The blister package will include an appropriate number of unit dosage forms in a sealed blister package with cardboard, paperboard, foil or plastic backing, and is encapsulated in a suitable lid. Each blister container can be numbered or otherwise marked, for example, starting from the 1st day.

[0189] In certain embodiments, the subject delivery system comprises an injection device. Exemplary non-limiting drug delivery devices include injection devices, such as pen injectors, and needle / syringe devices. Pen injectors are well known in the art. Exemplary devices that may be suitable for use in the methods of the present invention are any of the various pen injectors from Becton Dickinson, such as the BD TM Pen, BD TM PEN II, BD TM autoinjectors; pen injectors from Innoject, Inc.; any of the drug delivery pen devices discussed in U.S. Patent Nos. 5,728,074, 6,096,010, 6,146,361, 6,248,095, 6,277,099, and 6,221,053; the disclosures of each of which are incorporated herein by reference; etc. The drug delivery pen may be disposable, or reusable and refillable.

[0190] The present invention provides a delivery system for vaginal or rectal delivery of an active agent to the vagina or rectum of an individual. The delivery system includes a device for inserting into the vagina or rectum. In certain embodiments, the delivery system includes an applicator for delivering a preparation into the vagina or rectum; and a container containing a preparation containing an active agent. In these embodiments, the container (e.g., a tube) is suitable for delivering the preparation into the applicator. In other embodiments, the delivery system includes a device for inserting into the vagina or rectum, the device including an active agent. For example, the device is coated with, impregnated with, or contains a preparation containing an active agent.

[0191] In certain embodiments, the vaginal or rectal delivery system is a tampon or tampon-shaped device containing the subject preparation. Drug delivery tampons are known in the art, and any such tampons can be used in combination with the subject drug delivery system. Drug delivery tampons are described, for example, in U.S. Patent No. 6,086,909, the disclosure of which is incorporated herein by reference. If a tampon or tampon-shaped device is used, there are many ways to incorporate the active agent into the device. For example, the drug can be incorporated into the gelatinous bioadhesive reservoir at the tip of the device. As another option, the drug can be in the form of a powder material located at the tip of the tampon. The drug can also be absorbed into the fibers at the tip of the tampon, for example, by dissolving the drug in a pharmaceutically acceptable carrier and absorbing the drug solution into the fibers of the tampon. The drug can also be dissolved in a coating material that is applied to the tip of the tampon. As another option, the drug can be incorporated into an insertable suppository that is placed in association with the tip of the tampon.

[0192] In other embodiments, the drug delivery device is a vaginal or rectal ring. The vaginal or rectal ring is usually composed of an inert elastomeric ring, which is coated with another layer of elastomer containing the active agent to be delivered. The ring can be easily inserted, left in place for a desired period of time (e.g., up to 7 days), and then removed by the user. Alternatively, the ring may include a third outer rate-controlling elastomer layer that does not contain the drug. Alternatively, the third ring may contain a second drug for a dual release ring. The drug can be incorporated into polyethylene glycol throughout the silicone elastomer ring as a reservoir for the drug to be delivered.

[0193] In other embodiments, the subject vaginal or rectal delivery system is a vaginal or rectal sponge. As described in the literature, the active agent is incorporated into a silicone matrix that is coated on a cylindrical drug-free polyurethane sponge.

[0194] Vaginal suppositories, tablets, and suppositories are other examples of drug delivery systems that can be used, for example, to practice the methods of the present disclosure. These systems have been extensively described in the literature.

[0195] Bioadhesive microparticles constitute another drug delivery system suitable for use in the present invention. The system is a multiphase liquid or semisolid preparation that, like many suppository preparations, does not seep out of the vagina or rectum. These substances adhere to the vaginal or rectal walls and release the drug over a period of time. Many of these systems are designed for nasal use, but can also be used in the vagina or rectum (e.g., U.S. Patent No. 4,756,907, the disclosure of which is incorporated herein by reference). The system may include microspheres containing an active agent; and a surfactant for enhancing drug uptake. The microparticles have a diameter of 10 μm to 100 μm and can be prepared from starch, gelatin, albumin, collagen or dextran.

[0196] Another system is a container (e.g., tube) containing a subject formulation suitable for use with an applicator. The active agent is incorporated into creams, lotions, foams, pastes, ointments and gels, which can be applied to the vagina or rectum using an applicator. Methods for preparing medicaments in the form of creams, lotions, foams, pastes, ointments and gels can be found throughout the literature. An example of a suitable system is a standard fragrance-free lotion formulation containing glycerin, ceramides, mineral oil, petrolatum, parabens, spices and water, such as those sold under the trademark JERGENS TM (Andrew Jergens Co., Cincinnati, Ohio). Suitable nontoxic pharmaceutically acceptable systems for the compositions of the present invention are apparent to those skilled in the art of pharmaceutical formulation, and examples are described in Remington's Pharmaceutical Sciences, 19th edition, compiled by AR Gennaro, 1995. The choice of a suitable carrier will depend on the exact nature of the particular vaginal or rectal dosage form desired, e.g., whether the active ingredient is formulated as a cream, lotion, foam, ointment, paste, solution or gel, and the characteristics of the active ingredient. Other suitable delivery devices are described in U.S. Patent No. 6,476,079, the disclosure of which is incorporated herein by reference.

[0197] Treatment

[0198] The present disclosure also provides a method for treating a subject by administering one or more fatty acid β-hydroxy ester compounds and / or one or more butanediol esters or pharmaceutically acceptable salts thereof to the subject. In certain embodiments, the method includes administering one or more fatty acid β-hydroxy ester compounds or pharmaceutically acceptable salts thereof to the subject. In other embodiments, the method includes administering one or more butanediol esters or pharmaceutically acceptable salts thereof to the subject. In certain embodiments, the method includes administering a combination of one or more fatty acid β-hydroxy ester compounds or pharmaceutically acceptable salts thereof and one or more butanediol esters or pharmaceutically acceptable salts thereof to the subject. Wherein fatty acid β-hydroxy ester compounds and butanediol esters are both administered to the subject, and in certain cases, the mass ratio of fatty acid β-hydroxy ester compounds to butanediol esters ranges from 10: 1 to 1: 1, such as 9: 1 to 1.5: 1, such as 8: 1 to 2: 1, such as 7: 1 to 2.5: 1, such as 6: 1 to 3: 1 and including 5: 1 to 4: 1. In other cases, the mass ratio of fatty acid β-hydroxy ester compound to butanediol ester ranges from 1:10 to 1:1, such as 1:9 to 1:1, such as 1:8 to 1:1.5, such as 1:7 to 1:2, such as 1:6 to 1:2.5 and including 1:5 to 1:3.

[0199] In certain embodiments, the method comprises treating one or more of the following diseases: Alzheimer's disease, epilepsy, Parkinson's disease, heart failure, traumatic brain injury, stroke, hemorrhagic shock, acute lung injury after fluid resuscitation, acute kidney injury, myocardial infarction, myocardial ischemia, diabetes, glioblastoma multiforme, diabetic neuropathy, prostate cancer, amyotrophic lateral sclerosis, Huntington's disease, cutaneous T-cell lymphoma, multiple myeloma, peripheral T-cell lymphoma, HIV, Niemann-Pick C disease, age-related macular degeneration, gout, atherosclerosis, rheumatoid arthritis and multiple sclerosis by administering to the subject one or more fatty acid β-hydroxy ester compounds and / or one or more butanediol esters described herein.

[0200] In some embodiments, method includes treating Alzheimer's disease by applying one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters as described herein to the subject.In some cases, the amount (as described above) of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the subject is enough to reduce the epileptic activity of the subject.For example, the subject method may include applying a certain amount of subject fatty acid β-hydroxy ester compounds or butylene glycol esters, and the amount is enough to reduce epileptic activity by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99% and including reducing epileptic activity more than 99.9%.

[0201] In other cases, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the subject is sufficient to increase the subject's cognition. For example, the subject method may include administering an amount of subject fatty acid β-hydroxy ester compounds or fatty acid esters of butylene glycol, sufficient to increase the subject's cognition by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99% and including increasing the subject's cognition by more than 99.9%.

[0202] In other cases, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters administered to the subject is sufficient to reduce the rate of decline of cognition in the subject. For example, the subject method may include administering an amount of a subject fatty acid β-hydroxy ester compound or a fatty acid ester of butylene glycol, the amount being sufficient to reduce the rate of decline of cognition in the subject by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99% and including reducing the rate of decline of cognition in the subject by more than 99.9%.

[0203] Cognitive level in a subject may be assessed by any convenient protocol including, but not limited to, the Montreal Cognitive Assessment (MoCA), the Saint Louis University Mental State Examination (SLUMS), the Mini-Mental State Examination (MMSE), and for research purposes, the Alzheimer's Disease Assessment Scale, Cognitive (ADAS-Cog), and assessments including activities of daily living (ADL) and instrumental activities of daily living (IADL).

[0204] In certain embodiments, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters administered to a subject is sufficient to improve the subject's daily functioning, such as determined by assessments of activities of daily living (ADL) and instrumental activities of daily living (IADL).

[0205] In other embodiments, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the subject is sufficient to reduce agitation in the subject. For example, the subject method may include administering a certain amount of subject fatty acid β-hydroxy ester compounds or fatty acid esters of butylene glycol, sufficient to reduce agitation in the subject by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99% and including reducing agitation in the subject by more than 99.9%. Agitation can be assessed by any convenient scheme, for example, by a neuropsychiatric inventory (NPI) assessment.

[0206] In other embodiments, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters administered to a subject is sufficient to reduce delirium in the subject. For example, the subject method may include administering an amount of a subject fatty acid β-hydroxy ester compound or a fatty acid ester of butylene glycol sufficient to reduce delirium in the subject by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99%, and including reducing delirium in the subject by more than 99.9% in the subject.

[0207] In other embodiments, the amount of one or more fatty acid β-hydroxy ester compounds and / or one or more butylene glycol esters applied to the experimenter is enough to alleviate the pressure of the experimenter experienced by the caregiver.For example, the subject method can include giving a certain amount of subject fatty acid β-hydroxy ester compounds or the fatty acid ester of butylene glycol, being enough to reduce the pressure experienced by the caregiver by more than 5%, such as more than 10%, such as more than 15%, such as more than 25%, such as more than 40%, such as more than 50%, such as more than 75%, such as more than 90%, such as more than 95%, such as more than 99%, and be included in the experimenter so that the pressure experienced by the caregiver is reduced by more than 99.9%.The pressure of the caregiver can be assessed by any convenient scheme, for example, assessed by the perceived stress scale (PSS).

[0208] When implementing the subject method, the scheme of a specific subject can vary, for example, according to age, weight, severity of pain, general health of the subject, and the specific concentration of the fatty acid ester of the fatty acid β-hydroxy ester compound and / or butanediol administered. In an embodiment, the dosage delivered during administration can vary in some cases, ranging from 5mg to 800mg. Therefore, depending on the subject's physiology and the desired therapeutic effect, The dosage range provided by the subject method can be 5mg to 800mg, such as 10mg to about 500mg, such as 20mg to 400mg, such as 25mg to 350mg, such as 30mg to 300mg, such as 40mg to 250mg and including 40mg to 200mg.

[0209] Therefore, dosage interest can vary, and its scope is about 0.1g / kg~25g / kg / day, such as 0.1g / kg~20g / kg / day, such as 0.1g / kg~18g / kg / day, such as 0.1g / kg~15g / kg / day, such as 0.1g / kg~10g / kg / day, including 0.1g / kg~5g / kg / day, such as 0.5g / kg~10g / kg / day, such as 0.5g / kg~9g / kg / day, such as 0.5g / kg~8g / kg / day, such as 0.5g / kg~5g / kg / day, such as 0.5g / kg~3g / kg / day, such as 0.5g / kg~2g / kg / day and including 0.5g / kg~1g / kg / day. In some cases, dosage is 1g / kg / day. In other embodiments, the dosage range may be 0.1 g / kg to 6.5 g / kg four times / day (QID), such as 0.1 to 5 g / kg QID, such as 0.1 g / kg to 4 g / kg QID. In other embodiments, the oral dosage range may be 0.01 g / kg to 8.5 g / kg three times / day (TID), such as 0.1 g / kg to 6 g / kg TID, such as 0.1 g / kg to 5 g / kg TID, including 0.1 g / kg to 4 g / kg TID. In other embodiments, the oral dosage range may be 0.1 g / kg to 13 g / kg twice / day (BID), such as 0.1 g / kg to 12 g / kg BID, such as 5 g / kg to 10 g / kg BID, including 0.1 g / kg to 8 g / kg BID. The amount of compound administered will depend on the subject's physiology, the subject's absorptivity of the fatty acid β-hydroxy ester compound and / or the fatty acid ester of butanediol, and the magnitude of the desired therapeutic effect. Dosage regimens may include, but are not limited to, administration five times / day, four times / day, three times / day, twice / day, once / day, three times / week, twice / week, once / week, twice / month, once / month, and any combination thereof.

[0210] In certain embodiments, the subject methods may include chronic administration, which requires administration of the subject methods and compositions in multiple doses over a long period of time, for example, for more than one month and up to 10 years.

[0211] The duration between the dose intervals in the multiple dose interval treatment scheme can vary according to the physiology of the subject or the treatment scheme determined by the healthcare professional. In some cases, the duration between the dose intervals in the multiple dose treatment scheme can be predetermined and carried out at regular intervals. Therefore, the time between the dosing intervals can vary, and can be more than 0.5 hours, such as more than 1 hour, such as more than 2 hours, such as more than 4 hours, such as more than 8 hours, such as more than 12 hours, such as more than 16 hours, such as more than 24 hours, such as more than 48 hours and including more than 72 hours.

[0212] In certain embodiments, the subject method comprises administering to a subject a combination of one or more fatty acid β-hydroxy ester compounds or pharmaceutically acceptable salts thereof and / or one or more butylene glycol esters or pharmaceutically acceptable salts thereof and a ketogenic diet. The phrase "ketogenic diet" herein is used in its conventional sense to refer to a diet that provides ketone bodies as a primary energy source after consumption, digestion, and metabolism. Ketone bodies physiologically provided by a ketogenic diet include acetoacetate, β-hydroxybutyrate, and acetone. Suitable ketogenic diets may include, but are not limited to, the following descriptions: Fenton et al., ICAN 2009, 1: 338; Neal et al., Lancet Neurology 2008, 7: 500; Hartman and Vinning, Epilepsia 2007, 1: 31; Kossoff et al., Epilepsia 2009, 50: 304, the disclosures of which are incorporated herein by reference. In implementing the subject method, the subject fatty acid β-hydroxy ester compound or a pharmaceutically acceptable salt thereof and / or butanediol ester or a pharmaceutically acceptable salt thereof can be administered before, after, or together with the ketogenic diet. In certain embodiments, the subject method includes administering the subject fatty acid β-hydroxy ester compound or a pharmaceutically acceptable salt thereof and / or butanediol ester or a pharmaceutically acceptable salt thereof before starting the ketogenic diet. In certain embodiments, in other embodiments, the method includes administering the subject fatty acid β-hydroxy ester compound or a pharmaceutically acceptable salt thereof and / or butanediol ester or a pharmaceutically acceptable salt thereof after completing one or more intervals of the ketogenic diet. In other embodiments, the method includes administering the subject fatty acid β-hydroxy ester compound or a pharmaceutically acceptable salt thereof and / or butanediol ester or a pharmaceutically acceptable salt thereof together with the ketogenic diet.

[0213] Exemplary non-limiting aspects of the present disclosure

[0214] The various aspects (including embodiments) of the present subject matter described above may be beneficial alone or in combination with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the present disclosure numbered 1-48 are provided below. It will be apparent to those skilled in the art when reading this disclosure that each individually numbered aspect may be used or combined with any of the previously or subsequently individually numbered aspects. This is intended to provide support for the combination of all of these aspects, and is not limited to the combination of the aspects explicitly provided below.

[0215] 1. Compounds of formula I:

[0216]

[0217] in,

[0218] R1 is H or C(1-6) alkyl or alkyl having a substituent; and

[0219] R2 and R3 are independently unsubstituted or substituted C(4-30)alkyl.

[0220] 2. The compound according to 1, wherein R1 is an unsubstituted C(1-6) alkyl group.

[0221] 3. The compound as described in 2, wherein R1 is methyl.

[0222] 4. The compound according to any one of 1 to 3, wherein R2 and R3 are independently unsubstituted C(6-18) alkyl.

[0223] 5. The compound according to any one of 1 to 3, wherein R2 and R3 are independently unsubstituted C6 alkyl.

[0224] 6. The compound according to any one of 1 to 3, wherein R2 and R3 are independently unsubstituted C8 alkyl groups.

[0225] 7. The compound as described in 1, wherein the compound is a compound of formula Ia:

[0226]

[0227] in,

[0228] R1 is H or C(1-6) alkyl or alkyl having a substituent; and

[0229] R2 and R3 are independently unsubstituted or substituted C(4-30)alkyl.

[0230] 8. The compound according to 7, wherein R1 is an unsubstituted C(1-6) alkyl group.

[0231] 9. The compound according to 8, wherein R1 is methyl.

[0232] 10. The compound according to any one of 7 to 9, wherein R2 and R3 are independently unsubstituted C(6-18) alkyl.

[0233] 11. A compound as described in any one of 7 to 9, wherein R2 and R3 are independently C6 alkyl groups having no substituent.

[0234] 12. The compound according to any one of 7 to 9, wherein R2 and R3 are independently unsubstituted C8 alkyl groups.

[0235] 13. The compound of 1, wherein the compound is a compound of formula Ib:

[0236]

[0237] in,

[0238] R1 is H or C(1-6) alkyl or alkyl having a substituent; and

[0239] R2 and R3 are independently unsubstituted or substituted C(4-30)alkyl.

[0240] 14. The compound according to 13, wherein R1 is an unsubstituted C(1-6) alkyl group.

[0241] 15. The compound according to 14, wherein R1 is methyl.

[0242] 16. The compound according to any one of 13 to 15, wherein R2 and R3 are independently unsubstituted C(6-18) alkyl.

[0243] 17. The compound according to any one of 13 to 15, wherein R2 and R3 are independently unsubstituted C6 alkyl.

[0244] 18. The compound according to any one of 13 to 15, wherein R2 and R3 are independently unsubstituted C8 alkyl groups.

[0245] 19. A composition comprising the compound according to any one of 1 to 18 and a pharmaceutically acceptable carrier.

[0246] 20. A method comprising administering a therapeutically effective amount of the compound of any one of 1 to 18 or the composition of 19 to a subject in need thereof.

[0247] 21. The method of 20, wherein the therapeutically effective amount is sufficient to reduce epileptiform activity in the brain of the subject.

[0248] 22. A method of treating one or more of the following diseases: Alzheimer's disease, epilepsy, Parkinson's disease, heart failure, traumatic brain injury, stroke, hemorrhagic shock, acute lung injury after fluid resuscitation, acute kidney injury, myocardial infarction, myocardial ischemia, diabetes, glioblastoma multiforme, diabetic neuropathy, prostate cancer, amyotrophic lateral sclerosis, Huntington's disease, cutaneous T-cell lymphoma, multiple myeloma, peripheral T-cell lymphoma, HIV, Niemann-Pick C disease, age-related macular degeneration, gout, atherosclerosis, rheumatoid arthritis and multiple sclerosis, the method comprising:

[0249] A therapeutically effective amount of the compound of any one of 1 to 18 or the composition of 19 is administered to a subject.

[0250] 23. The method of 22, wherein the therapeutically effective amount is sufficient to reduce epileptiform activity in the subject's brain.

[0251] 24. A method of reducing epileptiform activity in the brain of a subject, the method comprising administering to the subject a therapeutically effective amount of the compound of any one of 1 to 18 or the composition of 19.

[0252] 25. A food supplement comprising the compound according to any one of 1 to 18.

[0253] 26. A composition comprising:

[0254] A food supplement comprising the compound of any one of 1 to 18; and

[0255] One or more other components of a ketogenic diet.

[0256] 27. The composition of 26, wherein the compound is present in the composition in an amount of about 1% w / w to about 25% w / w.

[0257] 28. The composition of 26, wherein the compound is present in the composition in an amount of about 5% w / w to about 15% w / w.

[0258] 29. The composition of 26, wherein the compound is present in the composition in an amount of about 10% w / w.

[0259] 30. The composition of 26, wherein the mass ratio of fat to protein to carbohydrates contained in the ketogenic diet is about 2:1 to about 10:1.

[0260] 31. The composition of 30, wherein the mass ratio of fat to protein to carbohydrates contained in the ketogenic diet is about 3:1 to about 6:1.

[0261] 32. The composition of 30, wherein the ketogenic diet comprises fat, protein, and carbohydrates in a mass ratio of about 4:1.

[0262] 33. Compounds of formula II:

[0263]

[0264] in,

[0265] R4 is H or C(1-6) alkyl or alkyl having a substituent; and

[0266] R5 and R6 are independently unsubstituted or substituted C(4-30)alkyl.

[0267] 34. The compound as described in 33, wherein R4 is a C(1-6) alkyl group having no substituent.

[0268] 35. The compound of 34, wherein R4 is methyl.

[0269] 36. The compound according to any one of 33 to 35, wherein R5 and R6 are independently unsubstituted C(6-18) alkyl.

[0270] 37. The compound according to any one of 33 to 35, wherein R5 and R6 are independently unsubstituted C6 alkyl.

[0271] 38. The compound according to any one of 33 to 35, wherein R5 and R6 are independently unsubstituted C8 alkyl groups.

[0272] 39. The compound of 33, wherein the compound is a compound of formula Ia:

[0273]

[0274] in,

[0275] R4 is H or C(1-6) alkyl or alkyl having a substituent; and

[0276] R5 and R6 are independently unsubstituted or substituted C(4-30) alkyl.

[0277] 40. The compound as described in 39, wherein R4 is a C(1-6) alkyl group having no substituent.

[0278] 41. The compound of 40, wherein R4 is methyl.

[0279] 42. The compound according to any one of 39 to 41, wherein R5 and R6 are independently unsubstituted C(6-18) alkyl.

[0280] 43. The compound according to any one of 39 to 41, wherein R5 and R6 are independently unsubstituted C6 alkyl.

[0281] 44. The compound according to any one of 39 to 41, wherein R5 and R6 are independently unsubstituted C8 alkyl.

[0282] 45. The compound of 33, wherein the compound is a compound of formula Ib:

[0283]

[0284] in,

[0285] R4 is H or C(1-6) alkyl or alkyl having a substituent; and

[0286] R5 and R6 are independently unsubstituted or substituted C(4-30) alkyl.

[0287] 46. ​​The compound as described in 45, wherein R4 is a C(1-6) alkyl group having no substituent.

[0288] 47. The compound of 46, wherein R4 is methyl.

[0289] 48. The compound according to any one of 45 to 47, wherein R5 and R6 are independently unsubstituted C(6-18) alkyl.

[0290] 49. The compound according to any one of 45 to 47, wherein R5 and R6 are independently unsubstituted C6 alkyl.

[0291] 50. The compound according to any one of 45 to 47, wherein R5 and R6 are independently unsubstituted C8 alkyl.

[0292] 51. A composition comprising the compound according to any one of 33 to 50 and a pharmaceutically acceptable carrier.

[0293] 52. A method comprising administering a therapeutically effective amount of a compound according to any one of 33 to 50 or a composition according to 51 to a subject in need thereof.

[0294] 53. The method of 52, wherein the therapeutically effective amount is sufficient to reduce epileptiform activity in the subject's brain.

[0295] 54. A method for treating one or more of the following diseases: Alzheimer's disease, epilepsy, Parkinson's disease, heart failure, traumatic brain injury, stroke, hemorrhagic shock, acute lung injury after fluid resuscitation, acute kidney injury, myocardial infarction, myocardial ischemia, diabetes, glioblastoma multiforme, diabetic neuropathy, prostate cancer, amyotrophic lateral sclerosis, Huntington's disease, cutaneous T-cell lymphoma, multiple myeloma, peripheral T-cell lymphoma, HIV, Niemann-Pick type C disease, age-related macular degeneration, gout, atherosclerosis, rheumatoid arthritis and multiple sclerosis, the method comprising:

[0296] A therapeutically effective amount of the compound of any one of 33 to 50 or the composition of 51 is administered to a subject.

[0297] 55. The method of 54, wherein the therapeutically effective amount is sufficient to reduce epileptiform activity in the subject's brain.

[0298] 56. A method of reducing epileptiform activity in the brain of a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of any one of 33 to 50 or a composition of 51.

[0299] 57. A food supplement comprising the compound of any one of 33 to 50.

[0300] 58. A composition comprising:

[0301] A food supplement comprising the compound of any one of 33 to 50; and

[0302] One or more components of a ketogenic diet.

[0303] 59. The composition of 58, wherein the compound is present in the composition in an amount of about 1% w / w to about 25% w / w.

[0304] 60. The composition of 58, wherein the compound is present in the composition in an amount of about 5% w / w to about 15% w / w.

[0305] 61. The composition of 58, wherein the compound is present in the composition in an amount of about 10% w / w.

[0306] 62. The composition of 58, wherein the ketogenic diet comprises fat, protein, and carbohydrates in a mass ratio of about 2:1 to about 10:1.

[0307] 63. The composition of 58, wherein the mass ratio of fat to protein and carbohydrates contained in the ketogenic diet is about 3:1 to about 6:1.

[0308] 64. A composition as described in 58, wherein the mass ratio of fat to protein and carbohydrates contained in the ketogenic diet is about 4:1.

[0309] Example

[0310] The following examples are presented to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. Standard abbreviations may be used, such as bp, base pair; kb, kilobase; pl, picoliter; s or sec, second; min, minute; h or hr, hour; aa, amino acid; kb, kilobase; bp, base pair; nt, nucleotide; im, intramuscular (m); ip, intraperitoneal (m); sc, subcutaneous (m); etc.

[0311] General synthetic procedure

[0312] The ketoesters described herein can be prepared by chemical synthesis schemes known to those skilled in the art (see, for example, Green et al., "Protective Groups in Organic Chemistry," (Wiley, 2nd ed. 1991); Harrison et al., "Compendium of Synthetic Organic Methods," Vol. 18 (John Wiley and Sons, 1971 1996); Beilstein Handbook of Organic Chemistry," Beilstein Institute of Organic Chemistry, Frankfurt, Germany; Feiser et al., "Reagents for Organic Synthesis," Vol. 117, (Wiley Interscience); Trost et al., "Comprehensive Organic Synthesis," (Pergamon Press, 1991); "Theilheimer's Synthetic Methods of Organic Chemistry," Vol. 145, (Karger, 1991); March, "Advanced Organic Chemistry," (Wiley Interscience, 1991; Larock "Comprehensive Organic Chemistry," Vol. 146, (Karger, 1991); Transformations," (VCH Publishers, 1989); Paquette, "Encyclopedia of Reagents for Organic Synthesis," (John Wiley & Sons, 1995), Bodanzsky, "Principles of Peptide Synthesis," (Springer Verlag, 1984); Bodanzsky, "Practice of Peptide Synthesis," (Springer Verlag, 1984). In addition, starting materials can be obtained from commercial sources or by recognized synthetic methods.

[0313] β-Hydroxy Ester Compounds

[0314] The β-hydroxy ester compounds described herein can be obtained by a synthetic route, generally as shown below:

[0315] Solution 1

[0316]

[0317] In Scheme 1, the hydroxyl group of β-hydroxy ester HE-1 is deprotonated with a weak base (e.g., pyridine) and reacted with a substituted acyl chloride to give acyl substituted β-hydroxy ester AHE-1. R4 can be H or a substituted or unsubstituted C(1-6) alkyl group; R5 and R6 are independently substituted or unsubstituted C(4-30) alkyl groups.

[0318] Solution 2

[0319]

[0320] In Scheme 2, sodium β-hydroxy ester HE-2 is reacted with an alkyl bromide in a polar aprotic solvent (e.g., dimethylformamide) to give alkyl β-hydroxy ester HE-3. The hydroxyl group of β-hydroxy ester KE-3 is deprotonated with a weak base (e.g., pyridine) and reacted with a substituted acyl chloride to give acyl substituted β-thienyloxy ester AHE-2. R4 can be H or a substituted or unsubstituted C(1-6) alkyl; R6 and R7 are independently substituted or unsubstituted C(4-30) alkyl.

[0321] Example 1 - Synthesis of Acyl-Substituted Ethyl β-Hydroxybutyrate

[0322] Solution 3

[0323]

[0324] As described in Scheme 3, 1 molar equivalent of R-ethyl β-hydroxybutyrate was dissolved in pyridine to a concentration of 0.2 molar. The solution was placed under a nitrogen atmosphere and cooled to 0°C. A substituted group (e.g., hexyl, Figure 1a-1b , and Hinkie, Figure 2a-2b ) was added dropwise to the β-hydroxy ester solution in 2 molar equivalents. The reaction mixture was warmed to room temperature and stirred overnight. The reaction was then charged to a separatory funnel and diluted with 1.5 volumes of ethyl acetate (based on the reaction volume), then washed 5 times with 0.5 volumes of hydrochloric acid solution (HCl 10%), then washed 4 times with 0.5 volumes of saturated sodium bicarbonate aqueous solution and once with brine. The ethyl acetate layer was then dried over magnesium sulfate, filtered and the solvent removed by rotary evaporation. 1 H NMR ( Figure 1a and 2a ) and gas chromatography mass spectrometry ( Figure 1b and 2b) analysis of the crude product. Pure (>95%) product was observed, the main contaminant being pyridine which was removed by further vacuum pumping.

[0325] Example 2 - Synthesis of Acyl-Substituted Hexyl β-Hydroxybutyrate

[0326] Solution 4

[0327]

[0328] As shown in Scheme 4, sodium β-hydroxybutyrate (1 molar equivalent) is suspended in dry dimethylformamide to a 0.2 molar concentration. 1-bromohexane (0.7 molar equivalent) is added, the reaction vessel is sealed and heated to 60°C with stirring for 18 hours, during which time the reaction mixture becomes clear. It is then cooled and charged to a separatory funnel and diluted with 1.5 volumes of ethyl acetate (based on the reaction volume). It is then washed five times with 0.5 volumes of saturated aqueous sodium bicarbonate solution and once with brine. The ethyl acetate layer is then dried over magnesium sulfate, filtered, and the solvent removed by rotary evaporation. 1 The crude product was analyzed by H NMR and GC-MS. Pure (>95%) product was observed, the major contaminant was 1-bromohexane which was removed by further vacuum pumping.

[0329] The hexyl β-hydroxybutyrate product is then dissolved in pyridine to a 0.2 molar concentration. The solution is placed under a nitrogen atmosphere and cooled at 0°C. The appropriate acid chloride (2 molar equivalents) is added dropwise to the solution via a syringe with stirring. The reaction mixture is warmed to room temperature and stirred overnight. The reaction mixture is then charged into a separatory funnel and diluted with 1.5 volumes of ethyl acetate (based on the reaction volume). It is then washed five times with 0.5 volumes of hydrochloric acid solution (10%), followed by four times with 0.5 volumes of saturated sodium bicarbonate aqueous solution, and once with brine. The ethyl acetate layer is then dried over magnesium sulfate, filtered, and the solvent removed by rotary evaporation. 1 H NMR ( Figure 3a ) and GC-MS( Figure 3b ) The crude product was analyzed. Pure (>95%) product was observed, the major contaminant was pyridine which was removed by further vacuum pumping.

[0330] 1,3-Butanediol ester

[0331] The 1,3-butanediol esters described herein can be obtained by the synthetic route generally shown below:

[0332] Solution 5

[0333]

[0334] In Scheme 5, the hydroxyl group of 1,3-butanediol BD-1 is deprotonated with a weak base (e.g., pyridine) and reacted with at least 2 equivalents of a substituted acyl chloride to give a homo-acyl substituted 1,3-butanediol ester BDE-1. R1 can be H or a substituted or unsubstituted C(1-6) alkyl group; R2 is a substituted or unsubstituted C(4-30) alkyl group.

[0335] Solution 6

[0336]

[0337] In Scheme 6, each hydroxyl group of 1,3-butanediol BD-1 is stepwise deprotonated with a weak base (e.g., pyridine) and reacted with 1 equivalent of a first substituted acid chloride and 1 equivalent of a second substituted acid chloride to obtain hetero-acyl substituted 1,3-butanediol ester BDE-2. R1 can be H or a substituted or unsubstituted C(1-6) alkyl group; R2 and R3 are independently substituted or unsubstituted C(4-30) alkyl groups.

[0338] Example 3 - Synthesis of Acyl-Substituted R-1,3-Butanediol

[0339] Solution 7

[0340]

[0341] As described in Scheme 7, R-1,3-butanediol (1 molar equivalent) was dissolved in pyridine to a 0.2 molar concentration. The solution was placed under a nitrogen atmosphere and cooled at 0°C. 3 to 3.5 molar equivalents of the appropriate acid chloride (hexyl, Figure 4a-4b ; Hinkie, Figure 5a-5b ) was added dropwise to the solution. The reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was then placed in a separatory funnel and diluted with 1.5 volumes (based on the reaction volume) of ethyl acetate. It was then washed five times with 0.5 volumes of hydrochloric acid solution (10%), then four times with 0.5 volumes of saturated sodium bicarbonate aqueous solution, and once with brine. The ethyl acetate layer was then dried over magnesium sulfate, filtered, and the solvent removed by rotary evaporation. 1 H NMR ( Figure 4a and 5a ) and GC-MS( Figure 4b and 5b ) The crude product was analyzed by HPLC. Pure (>95%) product was observed, the major contaminant was pyridine which was removed by further vacuum pumping.

[0342] Example 4 - Biological function of increasing BHB levels in the blood of wild-type C57BL / 6 male mice

[0343] Materials and methods

[0344] C6 and C8 esters of butanediol and C6 and C8 esters of β-hydroxybutyrate were synthesized as described above and purified for testing in wild-type C57BL / 6Ncrl male mice obtained from Charles River Laboratories. The mice were 8 months old at the time of the experiment. The biological functions of C6 and C8 esters of butanediol and β-hydroxybutyrate were tested by intraperitoneal injection at two doses of 50 μL and 100 μL. The injection was performed at about 9 am (the mice were kept on a 7a-7p light-dark cycle) and the mice were always able to obtain food and water. The molar amount injected was 0.13 to 0.41 millimoles, depending on the compound and the dose. The test mice weighed about 30 grams, so the amount of the compound injected was about 1 g / kg to 3 g / kg. In contrast, these amounts can theoretically provide about 1 / 20 of the daily caloric requirement of mice - equivalent to about 100 calories for humans. Blood was drawn by distal tail incision before injection (baseline) and 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours after injection. Approximately 40 μL of blood was collected into lithium-heparin microtiter tubes (Sarstedt CB-300LH), followed by centrifugation at 1500×G for 15 minutes at 4°C to separate plasma. Plasma BHB levels were determined by colorimetric enzyme assay (Stanbio Laboratory 2440-058).

[0345] result

[0346] All compounds increased blood BHB levels ( Figure 6a-6c At both doses tested, both C6-BHB and C8-BHB compounds increased BHB to levels seen on a ketogenic diet (0.5mM to 2mM) or to levels similar to BHB in mice after an overnight fast (1mM to 3mM) ( Figure 6a ). The only side effect observed was mild sedation at the higher doses of C6-BHB. Food intake was not measured, but a 6-hour fast did not significantly increase blood BHB levels ( Figure 6c ), so the increase seen with the compound was not due to inadvertent fasting. Importantly, the increase in blood BHB was greater than that seen with injection of a similar molar dose of BHB ( Figure 6c ) or a separate BD ( Figure 6b ), indicating that the fatty acid and BHB / BD components of the compound have separate and additive activities expected to increase blood BHB levels. Taken together, these data indicate that the novel BHB / BD-fatty acid ester compounds have the expected biological function of increasing blood BHB levels, in some cases to a similar extent as overnight fasting and for several hours.

[0347] Example 5 - Ketogenic diet rapidly suppresses epileptiform spikes in APPJ20 mice

[0348] Materials and methods

[0349] A ketogenic diet and overnight fasting were tested to determine the suppression of epileptiform peaks in one-year-old APPJ20 mice. APPJ20 mice carry the human APP gene, which has several mutations that cause Alzheimer's disease in humans. Heterozygous mice carrying the APP transgene were studied compared to non-transgenic (NTG) wild-type littermates. These mice exhibited early and severe cognitive deficits, as well as characteristic epileptiform peaks in the electroencephalogram (EEG) that led to their cognitive deficits. Reduction in epileptiform peaks caused by genetic manipulation or the anti-epileptic drug levetiracetam was associated with cognitive improvements. The studies described here used both male and female mice, and the data are shown as a summary of all mice unless mice were specifically differentiated by sex. Unless otherwise noted, there was no difference in the results when differentiated by sex. Mice and this epileptiform activity are further described in Palop et al., Neuron 2007, 55: 697-711.

[0350] A longitudinal cohort of mice with continuous 23-hour EEG recordings under three conditions was used in this study: first, baseline on a normal control diet; second, two days after starting a ketogenic diet (KD); and third, during an overnight fast. The mice were maintained on a control diet for eight weeks before the baseline EEG, and for three weeks between the KD and fasting EEG. The control diet was based on AIN-93M and included 10% calories from protein and 78% calories from carbohydrates. The ketogenic diet contained 90% calories from fat and zero carbohydrates, but was matched to the control diet on a per-calorie basis. The synthetic diet was custom-made to specifications by Harlan-Teklad (now Envigo). Caloric intake and blood BHB levels remained stable within two days of switching to the KD, and the KD and fasting produced similar 1mM to 2mM BHB blood levels.

[0351] During EEG recordings, mice moved freely in one of four clear plastic cylinders of the same size as their home cages. Harmonie Stellate software was used for EEG recording and automatic detection of spike peaks. Mice were also video recorded during EEG, and movement was quantified using Noldus Ethovision software. Prior to data analysis, raw movement data were cleaned to eliminate reflections or other artifacts. Gamma activity, defined as the power recorded by the EEG in the range of 20 Hz to 80 Hz, was quantified using LabChart software. Data analysis was performed using GraphPad Prism and a custom-written Perl program. EEG, data recording, and data analysis methods are further described in Verret et al., Cell 2012, 149:708-721.

[0352] result

[0353] Figure 7a-7h It was demonstrated that a ketogenic diet (but not fasting) consistently reduced epileptiform spikes in APPJ20 mice. As shown in Figure 7a, a 23-hour EEG recorded 2 days after starting the KD showed a mean peak reduction of approximately 30% from 2.13 spikes / minute to 1.48 spikes / minute compared to the previous baseline of the control diet. Mice that were fasted overnight showed no changes on average. Figure 7b The total number of spikes per hour during the 23-hour EEG recording is shown, indicating that spike suppression was consistent throughout the 23-hour recording. Figure 7c Shown are peak reductions for individual mice, normalized to baseline recordings for each mouse (solid circles, P<0.05; bars = median). Six of the nine mice had an overall reduction in peaks on KD, with none increasing above baseline. In contrast, while fasting reduced peaks in some mice, it increased peaks in others, resulting in no change on average.

[0354] Example 6 - Spike suppression on a ketogenic diet is not associated with interneuron function

[0355] Materials and methods

[0356] A ketogenic diet and overnight fasting were used to test the mechanism by which epileptiform spikes are suppressed by a ketogenic diet in one-year-old APPJ20 mice. The longitudinal cohort of mice that underwent continuous 23-hour EEG recordings is described above in Example 5. The methods of data acquisition and analysis are also described above in Example 5. Movement data, gamma power data, and spike data were collated in one-minute intervals to explore the temporal relationship between exploratory movements, inhibitory interneuron gamma activity, and spikes.

[0357] result

[0358] Exploratory movements are associated with suppression of epileptiform spikes through increases in inhibitory gamma activity from parvalbumin-positive interneurons (Verret et al., Cell 2012, 149:708-721). Three possible mechanisms by which KD may suppress spikes could be by increasing exploratory movements, exaggerating the induction of gamma activity from a given level of exploratory movement, or increasing baseline gamma activity. Overall movements were similar in all three conditions ( Figure 7d ), so increased exploratory activity did not explain the peak suppression with KD. The relative γ activity was also similar between the three conditions ( Figure 7e ). Figure 7f The best-fit linear regression line with 95% CI for the peak and movement per minute scatter plots is provided. APPJ20 mice generally had lower peaks with higher exploratory movements; KD was associated with peak suppression at all movement levels ( Figure 7f ). Figure 7g The best fit linear regression line with 95% CI for the scatter plot of normalized gamma activity per minute and exercise is provided. It shows that there was no change in the rate of induction of gamma activity by exercise on the KD. In other words, the rate of increase in gamma activity with exercise was similar between the control diet and the KD ( Figure 7g Finally, for most mice on KD, the overall mean level of gamma activity did not change and was not associated with changes in peak levels ( Figure 7h ). In summary, KD appears to act independently of parvalbumin-positive interneurons or downstream of presynaptic potentials that generate gamma activity in this neuronal population.

[0359] Example 7 - Long-term ketogenic diet reduces peaks and improves cognition

[0360] Materials and methods

[0361] The long-term persistence of the suppression of epileptiform peaks in KD and whether the peak suppression is associated with cognitive improvement was determined as follows. Groups of one-year-old APPJ20 mice (these mice are described in Example 5) were placed on a control diet or KD (diet is described in Example 5) and followed for three months. They received seven 50-minute EEG recording sessions in the second half of this period (EEG recording methods are described in Example 5). In addition, they were adapted to the open field in the first month, and two EEG sessions at two weeks and five weeks after adaptation served as probes to test whether familiarity with the previously adapted open field would reduce exploratory activity ( Figure 8a ).

[0362] Adaptation to the open field is a common test of visual-spatial memory and cognitive function (Verret et al., Cell 2012, 149: 708-721). Mice were placed in one of four identical transparent plastic chambers (40 cm × 40 cm × 30 cm) containing two beam arrays for measuring movement in the X and Y axes across the chamber floor; and another pair of arrays elevated in the Z axis for detecting rearing behavior. The apparatus was controlled by San Diego Instruments' Photobeam Activity System software. A custom program performed processing of the raw beam interruption data. Movement data from the open field were integrated with peak and gamma power data from the EEG using a custom program. Normal mice exhibited rapid adaptation in that their exploratory activity declined rapidly after repeated exposure to the open field over days to weeks. APPJ20 mice showed impaired adaptation in that exploratory activity remained elevated despite repeated exposure to the open field over days to weeks (Verret et al., Cell 2012, 149: 708-721).

[0363] result

[0364] Figure 8a-8h The reduction in epileptiform spikes due to KD was shown to persist for several months and correlated with cognitive improvements following adaptation to the open field. Figure 8a The experimental timeline is provided. The KD has been shown to suppress epileptiform spikes within days of starting the KD, and this effect will be evaluated to determine the sustainability of the diet over weeks to months and whether spike suppression is associated with cognitive improvements. Figure 8b showed that long-term KD consistently reduced epileptiform spikes by approximately 40% over three months, from a mean of 2.51 spikes / min to 1.53 spikes / min ( Figure 8b ). Figure 8c and 8d Displayed at all exercise levels ( Figure 8d ) and the entire 50-minute recording ( Figure 8c ), while mice on the control diet showed higher levels of peaks during the low-activity period following the recording period. As predicted, APPJ20 mice on the KD showed reduced exploratory activity when re-exposed to the open field after early adaptation. Overall locomotion levels were similar to those of non-transgenic (wild-type) controls ( Figure 8e and 8f ), more specific exploratory movements, such as movement through the center (relative to the periphery) of an open field ( Figure 8g ) and rear ( Figure 8h ). In contrast, APPJ20 transgenic mice fed a control diet continued to display hyperactivity and high levels of exploratory locomotion ( Figure 8e -h).

[0365] The previous 23-hour EEG study was longitudinal (Example 5), recording the same mice on a control diet and a KD, but this long-term study keeps mice in different dietary groups when necessary. In order to confirm that individual differences in peak levels between APPJ20 mice cannot explain the differences in diurnal rhythm peaks between dietary groups over days to weeks, a second longitudinal study was conducted. A group of mice was alternated from a control diet to a KD, and four 50-minute EEG recordings were obtained on each diet. Again, mice showed a significant suppression of epileptiform peaks on the KD, exceeding 50%, and also eliminated the relationship between exploratory activity and peaks seen previously.

[0366] Example 8 - Chronic Ketogenic Diet is Associated with Obesity, Male Survival, and Improved Learning

[0367] Materials and methods

[0368] Cognitive improvements, male survival, and adiposity from a chronic ketogenic diet were tested in one-year-old APPJ20 mice.

[0369] This 6-month study was initiated with 2-month-old APPJ20 mice (mice are further described in Example 5) and littermate non-transgenic (wild-type) control mice. The study answered three questions: 1) Does long-term feeding of mice on a KD have adverse metabolic effects, such as obesity; 2) Does the KD reduce the early mortality common in APPJ20 mice, which can be improved by other treatments that suppress epileptiform spikes; 3) Will APPJ20 mice on a KD improve in other tests of visual spatial cognition? The dietary intervention is described in Example 5.

[0370] The Morris water maze is a common test of visual-spatial memory. The maze consists of a shallow tub (122 cm in diameter) filled with water and rendered opaque by powdered white paint. Large, high-contrast visual cues were placed on the walls of the room. Mice were acclimated to the room and the pool one day before the start of the experiment. The experimental protocol consisted of six days of training (learning) trials, followed by an exploration (memory) trial 24 h after the final training trial. During the training trial, a 14 cm × 14 cm platform was submerged below the surface of the water. Repeated 60-second trials trained mice to find the hidden platform using visual cues in the room. The platform location remained constant during training, while the mice's entry point was changed semi-randomly between trials. On the final day, the platform was removed for a probe trial. Mouse movements were monitored using Ethovision video tracking software (Nolus). Performance in the training trial was assessed by the speed with which the mice located the platform during each trial. Performance in the exploration trial was assessed by the proportion of time the mice swam in the correct quadrant of the pool, where the platform had been previously located. After completion of this 7-day experimental protocol, the location of the platform was changed. The mice then underwent 3 days of "reversal training" to this new location before undergoing an exploration trial, with the platform removed 24 hours after the final training trial.

[0371] result

[0372] Figure 9a-9h showed that long-term ketogenic diet improved cognition as well as male survival. Figure 9a Shown are changes in body weight following KD or control diet for APPJ20 and NTG mice starting at 2 months of age. For APPJ20 and NTG mice, KD was substantially obesogenic, resulting in significant weight gain. Figure 9b Mean plasma BHB levels from six morning measurements taken approximately every two weeks from the start of the study are shown. Both groups developed plasma BHB levels averaging about 1 mM over 6 months, which were generally higher early on. These levels were about 10 times higher than mice on the control diet. Plasma glucose levels were similar in all groups. Males in all groups were heavier than females, and females had slightly higher BHB levels on the KD.

[0373] Fig.9c and 9d Survival curves for APPJ20 male and female mice on KD vs. control diet are provided. There were no deaths in NTG mice. Early mortality in APPJ20 mice was as high as 40%, which was considered fatal due to seizures. A trend toward reduced mortality was identified, which was shown by sex to be due to a significant reduction in the higher mortality of male mice; the already high survival of females was not affected ( Fig.9c and 9d ).

[0374] Figure 9e-9h The results of the Morris water maze test performed three months after the start of the diet are shown when the survivors were 5 months old. APP mice on the KD showed significantly improved performance in the hidden platform training (learning) phase of the water maze ( Fig.9e ). This improvement remained consistent when moving the platform position (reverse training, Figure 9g However, there were no differences in performance during the probe / remembering phase of the water maze either after initial hidden platform training or after reversal training ( Figure 9f and 9h ).

[0375] Example 9 - Ketogenic compounds inhibit the peak of normal diet

[0376] Materials and methods

[0377] C6 and C8 esters of butanediol and C6 and C8 esters of β-hydroxybutyrate were synthesized as described above. The efficacy of C6 ester of β-hydroxybutyrate in inhibiting the peak of APPJ20 mice was tested by intraperitoneal injection of APPJ20 mice (including male and female mice) aged 1 to 2 years. APPJ20 mice are described in Example 5.

[0378] Before and after intraperitoneal injection of C6-BHB or equal volume saline, the EEG of APPJ20 mice was recorded. After completing the first 50-minute recording, the injection was performed. The mice were then allowed to recover in their home cages for 20 minutes, followed by a second 50-minute EEG recording. Blood was drawn immediately after completing the second EEG phase to measure plasma BHB levels (as described in Example 4). The lower dose of the two previously tested doses from Example 4 was injected, 50 μL per 30 g body weight, which was about 1.5 g / kg and about 0.2 mmol.

[0379] During EEG recording, mice moved freely in a transparent cylinder approximately the size of their home cage. Methods for recording and analyzing EEG and for video tracking of movement are described in Example 5. Methods for blood draws and plasma BHB measurements are described in Example 4.

[0380] This study used a crossover design in which all mice were injected with C6-BHB and saline on different days with at least 48 h between injections. Data analysis was restricted to mice that completed all injections and EEG recordings.

[0381] result

[0382] Figures 10a-10f It was demonstrated that the compounds described herein that are metabolized to BHB can immediately reduce epileptiform peaks. Fig.10a Schematic diagrams of example ketogenic compounds having medium chain fatty acid esters linked to BHB are provided. Fig.10b Injection of C6-BHB was shown to increase blood BHB levels measured approximately 70 to 80 minutes after injection (after EEG).Injection of C6-BHB increased plasma BHB levels from a median of approximately 200 μM to a median of approximately 600 μM. Fig.10c The mean peak value of the 50-min EEG recording is shown in Figure 2. Fig.10d ) shows a consistent reduction after C6-BHB injection, which is similar to KD. C6-BHB reduced epileptiform spikes by approximately 35% compared to saline injection, from 1.25 spikes / min to 0.82 spikes / min. Analysis of spike reduction after C6-BHB, compared to saline, at the individual mouse level showed a significant reduction in most mice (solid circles, P<0.05; bars = median) ( Fig.10e ). The peak difference between C6-BHB and saline injections was most pronounced when mice were resting (and gamma activity was lowest), similar to KD ( Fig.10f ).

[0383] Summary of the results of Examples 6-9

[0384] Disruption of normal network activity and the associated epileptiform spikes of dysfunctional inhibitory interneurons are important for the pathogenesis of cognitive decline in Alzheimer's disease mouse models. Treatments that reduce epileptiform spikes can improve cognitive abilities in these models. Ketogenic diets and fasting have been used to treat certain forms of epilepsy, including those mechanisms associated with disrupted network activity in Alzheimer's disease models. For example, reduced expression of the sodium channel subunit SCN1A was found to be a key mechanism leading to early death, epileptiform spikes, and cognitive impairment in APPJ20 mice, and restoring normal expression improved these defects (Verret et al., 2012). Since SCN1A mutations cause a group of human genetic epilepsy, including Dravet syndrome, these diseases are usually refractory to conventional anti-epileptic drugs, but respond to ketogenic diet (KD) (Korff et al., J Child Neurol 2007, 22: 185). SCN1A mutant mice also respond to KD (Dutton et al., Epilepsia 2011, 52: 2050). Studies were conducted to determine whether KD could reduce epileptiform spikes and thereby improve cognition in a mouse model of Alzheimer's disease.

[0385] A ketogenic diet (but not fasting) consistently reduces epileptiform peaks in the APPJ20 Alzheimer's disease mouse model. This reduction in peaks is independent of or downstream of inhibitory interneuron function. The effect on peak reduction was sustained over several months of treatment. Long-term treatment resulted in cognitive improvements and open field adaptation in the water maze, and also improved survival in more severely affected male APPJ20 mice. Finally, treatment with a compound described herein that metabolizes to the ketone body β-hydroxybutyrate immediately reduced epileptiform peaks to a similar extent as a ketogenic diet. Provided herein are agents that increase blood levels of β-hydroxybutyrate or act on downstream targets of β-hydroxybutyrate to treat Alzheimer's disease by improving network function and improving epileptiform activity.

[0386] Example 10 - Pilot feeding study using diet supplemented with C6 esters of β-hydroxybutyrate

[0387] Materials and methods

[0388] The C6 monoester of β-hydroxybutyrate was synthesized as described above and purified for testing in mice. The C6 monoester of β-hydroxybutyrate was mixed into a ground control diet (10% calories from protein), the same control diet used in the above examples involving ketogenic and control diets. The food was placed in a glass jar in the animal cage at 19:00h at the beginning of the night feeding cycle. Different concentrations of C6 monoester of β-hydroxybutyrate (w / w) were mixed into the control diet to determine which concentration was sufficient to increase BHB levels in mice. n=4 mice / disease (2 mice / cage). All mice were 12-month-old C57BL / 6 males.

[0389] result

[0390] Fig.11 The intake of food containing different concentrations of C6 ester of β-hydroxybutyrate over time is depicted. It was observed that food containing 10 wt % and 20 wt % C6 monoester of β-hydroxybutyrate was less palatable to mice and was ingested at a lower rate. The addition of 5 wt % C6 monoester of β-hydroxybutyrate did not change the amount of food consumed by mice in the first 12 hours relative to the control.

[0391] Fig.12 Depicted is the weight loss of mice over time when fed a diet containing different amounts of a C6 monoester of β-hydroxybutyrate. The weight loss exhibited by mice fed a diet containing 2.5 wt% and 5 wt% of a C6 ester of β-hydroxybutyrate was not attributable to fasting, as the mice consumed normal amounts of food during the study.

[0392] Fig.13Blood glucose levels of mice during the feeding study are depicted. Mice that ingested a control food composition and mice that ingested a food with different amounts of C6 monoester of β-hydroxybutyrate exhibited blood glucose levels within the normoglycemic range (minimum = 70 mg / dL; maximum = 180 mg / dL). Mice that ingested a food with 10 wt % and 20 wt % of C6 monoester of β-hydroxybutyrate exhibited reduced blood glucose within the first 12 hours. The observations indicate that within the first 12 hours, these mice consumed small amounts of food with 10 wt % and 20 wt % of C6 monoester of β-hydroxybutyrate. Without wishing to be bound by any particular theory, it is believed that this fasting results in reduced blood glucose in these conditions.

[0393] Fig.14 Depicted are changes in blood concentrations of β-hydroxybutyrate over time after feeding mice a diet with different amounts of a C6 monoester of β-hydroxybutyrate. Diets containing 2.5 wt % or 5 wt % of a C6 monoester of β-hydroxybutyrate resulted in a slight increase in BHB levels in the blood, but not enough to provide levels above the target 500 μM threshold. Mice fed a diet containing 10 wt % or 20 wt % of a C6 monoester of β-hydroxybutyrate exhibited a greater increase in β-hydroxybutyrate concentrations. However, without being bound by any particular theory, it is believed that this increase in BHB levels may not be the result of a higher concentration of a C6 monoester of β-hydroxybutyrate in the diet, as the mice ate very little food. Instead, this increase may be explained by the mice being fasted.

[0394] Example 11 - Pilot feeding study using diets supplemented with esters of β-hydroxybutyrate and butylene glycol esters

[0395] Materials and methods

[0396] Different esters of β-hydroxybutyrate and butanediol were synthesized as described above and purified for testing in mice. The esters of β-hydroxybutyrate and butanediol esters supplemented to the mouse diet in this study were as follows: 1) C8 diester of butanediol (C8x2-BD); 2) C8 monoester of β-hydroxybutyrate (C8-BHB); 3) C6 diester of β-hydroxybutyrate (C6x2-BHB); 4) C6 monoester of β-hydroxybutyrate (C6-BHB); and 5) C6 diester of butanediol (C6x2-BD). Food compositions supplemented with 1,3-butanediol were also tested. The compound was mixed at 10% w / w in ground normal food (20% calories from protein) to match standard animal feed. Food was provided in glass jars in animal cages. n = 2 mice / disease, a total of 18 mice. All mice were 12-month-old C57BL / 6 males. Mice were placed in individual cages to more accurately record food intake. Food was switched back to normal chow at 72 hours (ground and in glass jars).

[0397] result

[0398] Fig.15 The intake of food containing the above listed butylene glycol esters and β-hydroxybutyrate over time is depicted. The control is normal food, no supplements. Foods supplemented with the following were also tested: 1,3-butylene glycol, 6-carbon fatty acid esters linked to 2-carbon alkyl (C6) as a control for C6 monoesters of β-hydroxybutyrate, and 6-carbon fatty acid esters linked to 6-carbon alkyl (C6x2) as a control for C6 diesters of β-hydroxybutyrate.

[0399] Depending on the compound, it took 2 to 4 days for the mice to adapt to the taste / smell of the supplemented food and begin consuming normal amounts.

[0400] Fig.16Describe when being supplemented with butanediol and beta-hydroxybutyrate different esters of food composition, mouse weight reduction over time.Results show, the compound causing significant weight loss is obviously different from the compound causing similar weight loss with control.Feeding the mouse of the diet of the C8 diester supplemented with butanediol loses about the same weight as the mouse of the diet consuming the control food composition, and consumes the C6 diester of beta-hydroxybutyrate, the C6 diester of butanediol, the C8 monoester of beta-hydroxybutyrate and the C6 monoester of beta-hydroxybutyrate of the mouse showing significant weight loss.Do not wish to be bound by any particular theory, it is believed that these results are not only explained by the difference of food intake (for example, the food intake of C6x2-BD is normal, but still causes weight loss).Not bound by any particular theory, it is believed that the weight loss of control mice can be explained by the following: lower food intake at the beginning of the experiment, while being adapted to the texture and placement of ground food, and using a single cage to cause higher heat loss.

[0401] Fig.17 Blood glucose levels of mice during the feeding study are depicted. Mice ingesting a control food composition and mice ingesting a food containing different types of esters of butanediol and beta-hydroxybutyrate exhibited blood glucose levels with a normoglycemic range (min = 70 mg / dL; max = 180 mg / dL). Some of the hypoglycemia exhibited can be explained by fasting (C8-BHB) and C6), but others (C6-BHB) may be a result of the compounds. No sedation was observed.

[0402] Fig.18 Depicted is the change in blood concentration of β-hydroxybutyrate over time after feeding mice a diet containing butanediol and different esters of β-hydroxybutyrate. The horizontal dashed line shows the target 500 μM threshold. When interpreting the results, fasting effects should be considered. For example, the high BHB values ​​of C8-BHB may be the result of fasting rather than the compound itself. A day / night pattern also became apparent after 48 hours, with higher BHB during night feeding and lower BHB during the day. Without wishing to be bound by any particular theory, it is believed that this diurnal pattern is consistent with ingested compounds being converted to BHB in vivo during feeding.

[0403] Example 12 - Mouse feeding study of diet supplemented with C6 diester of β-hydroxybutyrate

[0404] Materials and methods

[0405] Based on the results of the above pilot studies, the C6 diester of β-hydroxybutyrate was selected for a longer study, using a larger number of mice to see if it remained ketogenic even after normalization of food intake. The C6 diester of β-hydroxybutyrate was synthesized as described above and purified for mouse testing. The C6 diester of β-hydroxybutyrate was mixed at 10% w / w into a ground standard diet (20% calories from protein) which was re-pelleted. The re-pelleted diet was used to minimize any variable effects of the diet texture being changed to ground.

[0406] The mice consumed pelleted food after being placed in a glass jar in an animal cage. Four mice were tested for each diet, and the mice were caged individually. All mice were C57BL / 6 males, 12 months old. Daily body weight and caloric intake were monitored, and blood glucose and β-hydroxybutyrate levels were drawn on the eighth night. Mice fed with food supplemented with C6 esters of β-hydroxybutyrate initially ate less food and lost weight. By the seventh night, the weight of the mice was stable and the 24-hour caloric intake was similar between the control diet and the mice consuming food supplemented with C6 esters of β-hydroxybutyrate, the food. On the eighth night, the two groups of mice consumed similar calories in the four hours between the natural feeding time starting at 19:00h and the time of blood drawing starting at 23:00h.

[0407] result

[0408] Fig.19 Depicted is a comparison of plasma β-hydroxybutyrate, blood glucose, and caloric intake in mice fed a control diet versus mice fed a diet supplemented with 10% w / w of the C6 diester of β-hydroxybutyrate. Mice fed a diet supplemented with the C6 diester of β-hydroxybutyrate exhibited higher plasma concentrations of β-hydroxybutyrate, reaching the target threshold of 500 μM even as part of a normal diet with normal caloric intake.

[0409] Although the present invention has been described with reference to specific embodiments of the present invention, it will be appreciated by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the present invention. In addition, many modifications may be made to adapt specific circumstances, materials, compositions of matter, processes, process steps to the purpose, spirit and scope of the present invention. All of these modifications are within the scope of the appended claims.

Claims

1. An oral formulation comprising a compound of formula Ia: It in, R1 is methyl; and R2 and R3 are independently unsubstituted C5 alkyl groups.

2. Use of the oral formulation of claim 1 in the manufacture of a medicament for treating one or more diseases selected from the group consisting of Alzheimer's disease, epilepsy, Parkinson's disease, traumatic brain injury, stroke and Huntington's disease.

3. A composition comprising the oral preparation according to claim 1 and a pharmaceutically acceptable carrier.

4. Use of the oral formulation of claim 1 in the preparation of a medicament for reducing epileptiform activity in the brain of a subject.

5. A composition comprising: The oral preparation according to claim 1; and One or more additional components of a ketogenic diet, which is a diet that, after consumption, digestion, and metabolism, provides ketone bodies as the primary source of energy.

6. The composition according to claim 5, wherein The compound is present in the composition in an amount of 1% w / w to 25% w / w.

7. The composition according to claim 5 or 6, wherein The compound is present in the composition in an amount of 5% w / w to 15% w / w.

8. The composition according to claim 5 or 6, wherein The compound is present in the composition in an amount of 10% w / w.

9. The composition of claim 5, wherein The mass ratio of fat to protein to carbohydrates contained in the ketogenic diet is 2:1 to 10:

1.

10. The composition according to claim 9, wherein The mass ratio of fat to protein to carbohydrates contained in the ketogenic diet is 3:1 to 6:

1.

11. The composition according to claim 9 or 10, wherein The ketogenic diet contains fat, protein and carbohydrates in a mass ratio of 4:

1.

12. An oral preparation comprising the oral preparation according to claim 1, and a preservative and / or a flavoring agent.

13. The oral preparation according to claim 12, wherein The oral formulation comprises water.

14. The oral preparation according to claim 13, wherein The oral preparation is a syrup or elixir.

15. The oral preparation according to claim 12, wherein The oral formulation is a suspension.

16. The oral preparation according to claim 15, wherein The oral formulation further comprises a binder.

17. The oral preparation according to claim 16, wherein The binder is selected from the group consisting of cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, poly(ethylene glycol), sucrose, starch, and mixtures thereof.

18. The oral preparation according to claim 12, wherein The oral formulations contain flavoring agents.

19. The oral preparation according to claim 18, wherein The oral formulation comprises a flavoring selected from the group consisting of citric acid, menthol, glycine and orange powder.

20. The oral preparation according to claim 12, wherein The oral formulation contains a preservative.

21. The oral preparation according to claim 20, wherein The oral formulation comprises a preservative selected from the group consisting of sodium benzoate, sodium bisulfite, methylparaben, propylparaben, and mixtures thereof.

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