Prodrugs of indole-3-propionic acid and uses thereof

Abstract

Disclosed herein are compounds that are prodrugs of indole-3-propionic acid. Also disclosed herein are pharmaceutical compositions comprising the compounds, kits comprising the compounds, and methods of using the compounds in the treatment of disorders, including cardiovascular diseases.

Description

[0001] CCF-43944.601

[0002] PRODRUGS OF INDOLE-3-PROPIONIC ACID AND USES THEREOF

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 754,258, filed on February 5, 2025, the disclosure of which is incorporated herein by reference in its entirety.

[0004] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0005] This invention was made with government support under HL147823 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] FIELD

[0007] Provided herein are compounds that are prodrugs of indole-3-propionic acid. Also disclosed herein are pharmaceutical compositions comprising the compounds, kits comprising the compounds, and methods of using the compounds in the treatment of disorders, including cardiovascular diseases.

[0008] BACKGROUND

[0009] Tryptophan metabolites derived from gut microbes have recently been shown to be associated with metabolic disorders such as diabetes and steatohepatitis (Zhao et al. Exp Mol Med. 2019, 51:1-14; Abildgaard et al. Arch Physiol Biochem. 2018, 124:306-31). One specific tryptophan metabolite, indole-3-propionic acid (I3P or IPA, used interchangeably herein), has been associated in potentially treating an expanding list of different ailments including inflammation and cell oxidative damage (Wikoff et al. Proc Natl Acad Sei USA. 2009, 106:3698-703; Chyan et al. J Biol Chem. 1999, 274:21937-42; de Mello et al. Sei Rep 2017, 7:46337), colitis (Lee et al. Mol Pharmaceutics 2021, 18:1730-1741), skin disorders (WO 2020 / 185913 Al), and atherosclerosis and cardiovascular disease (Cason et al. J Vase Surg 2018, 68:1552-1562). Additional observed benefits of IPA include protection against influenza infection (Heumela et al. Gut Microbes 2024, 16, 2325067), protection in neurodegenerative disease models and cognitive impairment such as Alzheimer’s and Parkinson’s (Konopelski et al. Int. J. Mol. Sei. 2022, 23, 1222), protection against gut dysbiosis and endotoxin leakage (‘leaky gut’) by improvement in gut barrier function (Zhao 2019), and protection against heart failure of all forms, including heart failure with preserved ejection fraction (HFpEF) (Wang et al.CCF-43944.601

[0010] Circ. Res. 2024, 134, 371-389). T3P has also been reported to protect against development of non-alcoholic steatohepatitis (NASH) or Metabolic dysfunction-associated steatohepatitis (MASH) (Zhao 2019), and sepsis (Fang et al. Microbiol. Spectr. 2022, 10(3):e0012522; Huang et al. Front Cell Infect. Microbiol. 2022, 12:1015386; Wang et al. Mol. Med. 2023, 29(1):65).

[0011] A recent study showed that production of IPA was shown to be completely dependent on the presence of gut microflora (Wikoff 2009). It has been shown that dietary supplementation with IPA protects against many of the above listed disorders, including atherosclerosis in apolipoprotein E-deficient (ApoE- / -) mice by promoting macrophage reverse cholesterol transport (Xue et al. Circulation Research 2022, 131:404-420).

[0012] IPA appears to also play a role in protection against obesity and diabetes in obese patients with type 2 diabetes where IPA levels are significantly decreased (Jennis et al.

[0013] Neurogastroenterol Motil 2018, 30, el 3178). Furthermore, a recent study in cells and animals showed that IPA displays numerous anti-cancer activities. For example. IPA has been reported to foster cytostatic properties in breast cancer (Sari et al. Cancers 2020, 12, 2411) and other cancers (e.g. ovarian, triple negative breast cancer, colon cancer). In another study, IPA treatment was shown to promote nerve regeneration and repair (Serger et al. Nature 2022, 607, 585-592).

[0014] Other studies have shown further beneficial aspects of IPA where IPA has been reported to: 1) protect the brain from disease and oxidative damage; 2) inhibit liver fibrosis and lipotoxicity; 3) protect the lungs from bacterial and fungal infections; and 4) promote muscle growth and reduce inflammation (liang et al. Nutrients 2023, 15, 151).

[0015] SUMMARY

[0016] In one aspect, disclosed herein is compound of formula (I):

[0017]

[0018] or a pharmaceutically acceptable salt thereof, wherein:

[0019] A is an oligosaccharide or polysaccharide; and

[0020] n is at least 1.CCF-43944.601

[0021] In some embodiments, A is a cyclodextrin selected from a-cyclodextrin, P-cyclodextrin, and y-cyclodcxtrin. In some embodiments, the cyclodextrin is a-cyclodextrin. In some embodiments, the cyclodextrin is P-cyclodextrin. In some embodiments, the cyclodextrin is y-cyclodextrin. In some embodiments, A is selected from inulin, dextran, and dextrin. In some embodiments, A is inulin.

[0022] In some embodiments, the oligosaccharide or polysaccharide is further functionalized with one or more substituents independently selected from the group consisting of alkyl groups, hydroxyalkyl groups, and amino acids.

[0023] In some embodiments, n is 1-100, 1-50, 1-30, or 1-10.

[0024] In some embodiments, the compound of formula (I) has a calculated polar surface area of greater than 400 and a cLogP of less than zero.

[0025] In another aspect, disclosed herein is a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0026] In another aspect, disclosed herein is a method of treating a disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0027] In some embodiments, the disorder is selected from cardiovascular diseases. In some embodiments, cardiovascular disease is selected from heart failure, coronary heart disease, cor pulmonale, congenital heart defect, cardiomyopathy, myocardial infarction, abdominal aortic aneurysm, valvular heart disease, arrhythmia, peripheral arterial disease, cerebrovascular accident, atherosclerosis, and rheumatic heart disease. In some embodiments, the heart failure is heart failure with preserved ejection fraction (HFpEF). In some embodiments, the disorder is selected from obesity, diabetes, endotoxin leakage, cancer, inflammation, cell oxidative damage, skin disorders, influenza infection, neurodegenerative diseases, nonalcoholic steatohepatitis, and sepsis. In some embodiments, the disorder is a cancer selected from triple-negative breast cancer, colon cancer, glioblastoma, ovarian cancer, and leukemia. In some embodiments, the disorder is type II diabetes. In some embodiments, the disorder is intestinal endotoxin leakage resulting in steatohepatitis. In some embodiments, the steatohepatitis comprises metabolic dysfunction-associated steatohepatitis.CCF-43944.601

[0028] In some embodiments, the method further comprises administering a second therapeutic agent to the subject. In some embodiments, the second therapeutic agent is selected from antimicrobials, agents that improve intestinal motility, cardiovascular drags, a nicotinamide adenine dinucleotide (NAD)-boosting compound; and / or an a-amino-P-carboxymuconate-s-semialdehyde decarboxylase (ACMSD) inhibitor In some embodiments, the second therapeutic agent is selected from Omega 3 oil, salicylic acid, dimethylbutanol, garlic oil, olive oil, krill oil, Co enzyme Q-10, a probiotic, a prebiotic, dietary fiber, psyllium husk, bismuth salts, phytosterols, grape seed oil, green tea extract, vitamin D, antioxidants, turmeric, curcumin, and / or resveratrol. In some embodiments, the second therapeutic agent is a chemotherapeutic agent.

[0029] In another aspect, disclosed herein is a method of increasing levels of indole-3-propionic acid in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0030] In another aspect, disclosed herein is method of promoting health, longevity, vitality, vigor, and / or wellness in a subject, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0031] In another aspect, disclosed herein is a method of providing radioprotective effects to a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound of formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is undergoing radiation therapy for treatment of cancer.

[0032] Other aspects and embodiments will become apparent in light of the following description.

[0033] BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows data demonstrating the association of indole-3-propionic acid (I3P) with different phenotypes. Elevated I3P levels were inversely associated with increased risks for fastgrowing aneurysm, need for surgical intervention, all-cause mortality, MACE or Ml / stroke in CKD subjects, obesity, diabetes, abdominal aortic aneurysm, heart failure (HFrEF), and history of MI.CCF-43944.601

[0034] FIGS. 2A-B show data demonstrating the association of plasma I3P with abdominal aortic aneurysm (AAA) in the Cleveland AAA case / control Cohort (N=604); AAA is defined as having a baseline infrarenal aortic diameter >3.0 cm. FIG. 2A: Box-Whisker plots of I3P levels stratified by AAA status, Data are represented as boxplots: middle line is the median, the lower and upper boundaries to the boxes represent 25th and 75th percentiles, and the whiskers represent 5th and 95th percentile. P-value was calculated by Wilcoxon rank sum test. FIG. 2B: Forest plots indicating the odds of AAA according to the tertiles of I3P levels, multivariable logistic regression model for odds ratio included adjustments for age, sex, current smoking, hypertension, diabetes mellitus, HDL, LDL. Symbols represent odds ratios and the 95% confidence interval is indicated by line length.

[0035] FIGS. 3A-3D show data demonstrating the association of I3P levels with fast-growing aneurysm and need for surgical repair in the Cleveland Cohort. Fast-growing aneurysm was defined as AAA growth rate >4 mm / year, and need for surgical repair is defined as follow-up abdominal aortic diameter (AAD) >5.5 or AAA growth rate >4 mm / year. FIGS. 3A-3B: Kaplan-Meier plot depicting the risk for fast-growing aneurysm and need for surgical repair stratified by tertiles of I3P levels. FIGS. 3C-3D: Forest plots illustrating the risk of fast-growing aneurysm and need for surgical repair according to tertiles of I3P. Hazard ratios were calculated by Cox modelling using adjustments for age, sex, HDL, LDL, current smoking, hypertension, and diabetes mellitus. Symbols represent hazard ratios, and the 95% confidence interval is indicated by line length.

[0036] FIGS. 4A-4B show: (A) Plasma kinetics of Indole- 3 -propionic acid (I3P) in mice administered as a gastric gavage of either free I3P (red) or as a prodrug form of I3P (conjugates of inulin (black) or gamma-cyclodextrin (blue)). (B) Recovery of total I3P in feces (post base hydrolysis) from I3P gavaged mice. Shown are box- whisker plots of the total I3P measured in feces recovered from mice treated with each of the indicated ester conjugate forms of I3P (gamma-cyclodextrin I3P, and inulin-I3P) compared to free I3P.

[0037] FIG. 5 shows data from an ad libitum feeding study. Inulin IPA ester conjugate was mixed into either a normal chow diet (left panel) or a high fat (HF) diet (right panel) and fed to mice. Shown are monitored plasma I3P levels observed at the indicated times (baseline vs 1,4,7,10 or 14 days). Note that ad lib feeding of mice to I3P inulin prodrug form resulted in a steady increase in circulating I3P levels with both diets (Chow and HF).CCF-43944.601

[0038] DETAILED DESCRIPTION

[0039] Ongoing research continues to emphasize that human systemic levels of IPA have an association with several diseases (FIG. 1). Additionally, as demonstrated herein, previous observations (Wikoff 2009) have been confirmed, that the systemic absorption of orally-ingested indole-3-propionic acid is very rapid in a mouse model but is also quickly excreted and / or metabolized. This is not surprising given that IPA possesses pharmacokinetic properties that would favor absorption: 1) molecular weight < 400, 2) cLogP < 3, and 3) total polar surface area < 120 (Bergstrom et al. Eur J Pharm Sci 2014, 57, 173-199). A similar pharmacokinetic profile of IPA is anticipated to be observed in humans. Elevating IPA levels in a patient with AAA through supplementation, using a delayed-release form of IPA, may provide a different treatment option.

[0040] To arrive at a slow-release formulation of indole-3 -propionic acid, a conjugate compound was prepared, having a high polar surface area to disfavor systemic absorption in addition to having a cleavable linkage to release IPA, preferably in the colon where it is normally produced by gut microbes (George et al. PLoS ONE 2018; 13(9): e0203567). In some embodiments, the cleavable linkage is an ester linkage, which relies on an esterase to break the ester bond to the IPA-conjugated carrier and subsequently releasing free IPA. In other embodiments, the cleavable linkage is an amide linkage or a carbamate linkage. The conjugate compounds include compounds of Formula (I), described further below.

[0041] Definitions

[0042] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, any nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those that are well known and commonly used in the art. The meaning and scope of the terms should be clear; in the event, however of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0043] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise.CCF-43944.601

[0044] As used herein, the term “and / or” includes any and all combinations of listed items, including any of the listed items individually. For example, “A, B, and / or C” encompasses A, B, C. AB, AC, BC, and ABC, each of which is to be considered separately described by the statement “A, B, and / or C.”

[0045] For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.

[0046] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75thEd., inside cover, and specific functional groups are generally defined as described therein.

[0047] Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Sorrell, Organic Chemistry, 2ndedition, University Science Books, Sausalito, 2006; Smith, March’s Advanced Organic Chemistry: Reactions, Mechanism, and Structure, 7thEdition, John Wiley & Sons, Inc., New York, 2013; Larock, Comprehensive Organic Transformations, 3rdEdition, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3rdEdition, Cambridge University Press, Cambridge, 1987; the entire contents of each of which are incorporated herein by reference.

[0048] As used herein, the term “alkyl” refers to a radical of a straight or branched saturated hydrocarbon chain. The alkyl chain can include, e.g., from 1 to 24 carbon atoms (C1-C24 alkyl), 1 to 16 carbon atoms (C1-C16 alkyl), 1 to 14 carbon atoms (C1-C14 alkyl), 1 to 12 carbon atoms (Ci-C12 alkyl), 1 to 10 carbon atoms (C1-C10 alkyl), 1 to 8 carbon atoms (Ci-Cs alkyl), 1 to 6 carbon atoms (Ci-Ce alkyl), 1 to 4 carbon atoms (C1-C4 alkyl), 1 to 3 carbon atoms (C1-C3 alkyl), or 1 to 2 carbon atoms (C1-C2 alkyl). Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl.

[0049] As used herein, the term “amino acid” refers to any and all amino acids, including naturally occurring amino acids (e.g., a-amino acids), unnatural amino acids, modified aminoCCF-43944.601

[0050] acids, and non-natural amino acids. Tt includes both D- and L-amino acids. Natural amino acids include those found in nature, such as, e.g., the twenty-three amino acids that combine into peptide chains to form the building-blocks of a vast array of proteins. These are primarily L stereoisomers, although a few D-amino acids occur in bacterial envelopes and some antibiotics. Natural amino acids include those found in nature, such as, e.g., the 23 amino acids that combine into peptide chains to form the building-blocks of a vast array of proteins. These are primarily L stereoisomers, although a few D-amino acids occur in bacterial envelopes and some antibiotics. The “non-standard” natural amino acids include, for example, pyrrololysine (found in methanogenic organisms and other eukaryotes), selenocysteine (present in many non-eukaryotes as well as most eukaryotes), and N-formylmethionine (encoded by the start codon AUG in bacteria, mitochondria, and chloroplasts). “Unnatural” or “non-natural” amino acids are non-proteinogenic amino acids (e.g., those not naturally encoded or found in the genetic code) that either occur naturally or are chemically synthesized, examples of which include p-amino acids (P3and p2). homo-amino acids, proline and pyruvic acid derivatives, 3-substituted alanine derivatives, glycine derivatives, ring-substituted phenylalanine and tyrosine derivatives, linear core amino acids, diamino acids, D-amino acids, alpha-methyl amino acids, and N-methyl amino acids. Unnatural or non-natural amino acids also include modified amino acids. “Modified” amino acids include amino acids (e.g., natural amino acids) that have been chemically modified to include one or more chemical moieties not naturally present on the amino acid. For the most part, the names of naturally occurring and non-naturally occurring aminoacyl residues used herein follow the naming conventions suggested by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature as set out in “Nomenclature of a- Amino Acids (Recommendations, 1974)” Biochemistry, 14(2), (1975). Unless naturally occurring amino acids are referred to by their full name herein (e.g., alanine, arginine, etc.), they are designated by their conventional three-letter or single-letter abbreviations (e.g., Ala or A for alanine, Arg or R for arginine, etc.). In the case of less common or non-naturally occurring amino acids, unless they are referred to by their full name (e.g. sarcosine, ornithine, citrulline, aminoadipic acid, etc.), frequently used three- or four-character codes are employed for residues thereof, including, Sar or Sarc (sarcosine, i.e. N-methylglycine), Aib (a-aminoisobutyric acid). Dab (2,4-diaminobutanoic acid), Dapa (2,3-diaminopropanoic acid), y-Glu (y-glutamic acid), Gaba (y-aminobutanoic acid), P-ProCCF-43944.601

[0051] (pyrrolidine-3-carboxylic acid), and 8Ado (8-amino-3,6-dioxaoctanoic acid), Abu (2-amino butyric acid), phPro (P-homoproline), phPhe (P-homophenylalanine) Bip (P,p diphenylalanine), and Ida (Iminodiacetic acid).

[0052] As used herein, the term “hydroxy” refers to an -OH group.

[0053] As used herein, the term “hydroxyalkyl” refers to an alkyl group, as defined herein, in which at least one hydrogen atom is replaced with a hydroxy group. Representative examples of haloalkyl include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, and 3-hydroxypropyl.

[0054] As used herein, the term “oligosaccharide” refers to a carbohydrate compound having from 3 to 10 monosaccharide moieties, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 monosaccharide moieties. An oligosaccharide can be linear, cyclic, or branched.

[0055] As used herein, the term “polysaccharide” refers to a carbohydrate compound having more than 10 monosaccharide moieties. A polysaccharide can be linear, cyclic, or branched.

[0056] As used herein, in chemical structures the indication:

[0057]

[0058] represents a point of attachment of one moiety to another moiety (e.g., a substituent group to the rest of the compound).

[0059] For compounds described herein, groups and substituents thereof may be selected in accordance with permitted valence of the atoms and the substituents, such that the selections and substitutions result in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.

[0060] When substituent groups are specified by their conventional chemical formulae, written from left to right, such indication also encompass substituent groups resulting from writing the structure from right to left. For example, if a bivalent group is shown as -CH2O-, such indication also encompasses -OCH2-; similarly, -OC(O)NH- also encompasses -NHC(O)O-.

[0061] The terms “administer,” “administering,” or “administration,” as used herein refer to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing a compound or a pharmaceutical composition.

[0062] As used herein, the terms “condition,” “disease,” and “disorder” are used interchangeably.

[0063] An “effective amount” of a compound or composition refers to an amount sufficient to elicit a desired biological response (e.g., treating a condition). As will be appreciated by thoseCCF-43944.601

[0064] skilled in the art, the effective amount of a compound may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. An effective amount encompasses therapeutic and prophylactic treatment. For example, in treating cancer, an effective amount of a compound or composition may reduce tumor burden or stop the growth or spread of a tumor.

[0065] A “therapeutically effective amount” of a compound or composition is an amount sufficient to provide a therapeutic benefit in the treatment of a condition, or to delay or minimize one or more symptoms associated with the condition. In some embodiments, a therapeutically effective amount is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to minimize one or more symptoms associated with the condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment of the condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of the condition, or enhances the therapeutic efficacy of another therapeutic agent.

[0066] A “subject” to which administration is contemplated includes, but is not limited to, a human (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) and / or other non-human animals, for example, mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs) and birds (e.g., commercially relevant birds such as chickens, ducks, geese, and / or turkeys).

[0067] As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of. or inhibiting the progress of a disease or condition, or one or more signs or symptoms thereof. In some embodiments, “treatment,” “treat,” and “treating” require that signs or symptoms of the disease disorder or condition have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease or condition. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g.. in light of a history of symptomsCCF-43944.601

[0068] and / or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.

[0069] Compounds

[0070] Disclosed herein are compounds of formula (I):

[0071]

[0072] and pharmaceutically acceptable salts thereof, wherein:

[0073] A is an oligosaccharide or polysaccharide; and

[0074] n is at least 1.

[0075] In formula (I), the group A is an oligosaccharide or a polysaccharide. The oligosaccharide or polysaccharide can include varying lengths of monosaccharide units including, for example, D-glucose, D-fructose, D-galactose, L-galactose, D-mannose, L-arabinose, D-xylose. glucuronic acid, iduronic acid, D-glucosamine, D-galactosamine, N-acetylneuraminic acid, N-acetylmuramic acid, N-acetylglucosamine, and / or N-acetylgalactosamine, or the like.

[0076] In some embodiments, A is selected from a cyclodextrin, inulin, dextran, and dextrin. In some embodiments, A is a cyclodextrin, which is a cyclic oligosaccharide. In some embodiments, the cyclodextrin is selected from a-cyclodextrin, -cyclodextrin, and y-cyclodextrin. In some embodiments, the cyclodextrin is a-cyclodextrin. In some embodiments, the cyclodextrin is P-cyclodextrin. In some embodiments, the cyclodextrin is y-cyclodextrin. In some embodiments, A is inulin. In some embodiments. A is dextran. In some embodiments, A is dextrin.

[0077] In some embodiments, A is an oligosaccharide or a polysaccharide that is further functionalized with one or more substituent groups selected from alkyl groups, hydroxyalkyl groups, and amino acids. For example, in some embodiments, A is an oligosaccharide or polysaccharide (e.g.. a cyclodextrin, inulin, dextran, or dextrin) that is functionalized with one or more alkyl groups independently selected from methyl, ethyl, and isopropyl. In some embodiments, A is an oligosaccharide or polysaccharide (e.g., a cyclodextrin, inulin, dextran, orCCF-43944.601

[0078] dextrin) that is functionalized with one or more hydroxyalkyl groups independently selected from 2-hydroxyethyl and 3-hydroxypropyl. In some embodiments, A is an oligosaccharide or polysaccharide (e.g.. a cyclodextrin, inulin, dextran, or dextrin) that is functionalized with one or more amino acids independently selected from glycine, alanine, valine, leucine, isoleucine, lysine, arginine, histidine, proline, serine, threonine, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, cysteine, and methionine.

[0079] In formula (I), n is at least 1. This value represents the number of I3P moieties, i.e.,

[0080]

[0081] polysaccharide. As oligosaccharides and polysaccharides contain multiple primary hydroxy groups, the compounds are susceptible to esterification by activated forms of IPA. The degree of esterification (i.e., the value of n) will depend on the reaction stoichiometry used to prepare the compound of formula (I). Additionally, depending on the choice of group A, the maximum number that n can represent will vary. For example, when A is an oligosaccharide having three monosaccharide moieties, n may be 1, 2, or 3. In some embodiments, n is 1-100, 1-90, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15. 1-14, 1-11, 1-10, 1-9. 1-8, 1-7. 1-6, 1-5, 1-4, 1-3, or 1-2. In some embodiments, n is 1-30. In some embodiments, n is 1-10. In some embodiments, n is 1-10. In some embodiments, n is 1-5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23. 24. 25. 26. 27. 28, 29, or 30.

[0082]

[0083] oligosaccharide or polysaccharide A via an ester linkage, in which each groupCCF-43944.601

[0084]

[0085] covalently bound to a primary 6-OH group of a monosaccharide unit in the oligosaccharide or polysaccharide.

[0086] Those skilled in the art will appreciate that when compounds of formula (1) are synthesized by conjugating indole-3-propionic acid to the oligosaccharide or polysaccharide, a composition will be formed containing a plurality of compounds of formula (I), which may have

[0087] differing numbers of I3P moieties attached (i.e., groups

[0088]

[0089] composition comprising compounds of formula (I) may be defined by the weight percentage that corresponds to 13P moieties, which can be determined, e.g., by hydrolyzing the oligosaccharide / polysaccharide I3P ester and quantifying the amount of released free I3P mass spectrometry. In some embodiments, disclosed herein is a composition comprising compounds of formula (I), in which the weight percentage of IPA in the composition is about 10 wt% to about 50 wt%, or about 25 wt% to about 40 wt%, e.g., about 25 wt%, about 26 wt%, about 27 wt%, about 28 wt%, about 29 wt%, about 30 wt%, about 31 wt%, about 32 wt%, about 33 wt%, about 34 wt%, about 35 wt%, about 36 wt%, about 37 wt%, about 38 wt%, about 39 wt%, or about 40 wt%, as determined by mass spectrometry.

[0090] Oligosaccharides and polysaccharides such as cyclodextrins, inulin, dextrins, or dextrans have large calculated polar surface areas. The lipophilic character (cLogP) of these oligo- and poly-saccharides will also be very low. Specifically, the calculated polar surface areas for alpha, beta, and gamma cyclodextrins are >400 and the calculated LogP (cLogP) values are much less than zero (based on calculations using ChemDraw software version 18.0). The inulin-IPA conjugate would be expected to have a much higher polar surface area given that the general polysaccharide chain is much larger than the cyclodextrins. IPA esters of starch and modified starch would also be expected to possess very large polar surface areas and would be suitable conjugates for oral administration. Thus. IPA esters of these carriers would be expected toCCF-43944.601

[0091] possess high polar surface areas and very low (negative) cLogP values and would therefore be disfavored to undergo systemic absorption upon oral administration. Calculated polar surface areas and cLogP values of some of these esters are exemplified in Table 1. As observed in Table 1, the calculated polar surface areas of cyclodextrins having 4 IPA substitutions are at least 10-fold higher than IPA itself. The cLogP values are also much lower than IPA itself.

[0092] Table 1: cLogP and Polar Surface Areas of IPA ester conjugates

[0093]

[0094] CCF-43944.601

[0095]

[0096] CCF-43944.601

[0097] The compounds of the present disclosure have at least one asymmetric center. Additional asymmetric centers may be present depending upon the nature of the various substituent groups. Compounds with asymmetric centers give rise to enantiomers (optical isomers), diastereomers (configurational isomers) or both, and it is intended that all of the possible enantiomers and diastereomers, in mixtures and as pure or partially purified compounds, are included within the scope of this disclosure.

[0098] The independent syntheses of the enantiomerically or diastereomerically enriched compounds, or their chromatographic separations, may be achieved as known in the art by appropriate modification of the methodology disclosed herein. The absolute stereochemistry of a compound may be determined by using X-ray crystallography to determine the crystal structure of crystalline products or crystalline intermediates that are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration.

[0099] If desired, racemic mixtures of the compounds may be separated so that the individual enantiomers are isolated. The separation can be earned out by methods well known in the art. such as the coupling of a racemic mixture of compounds to an enantiomerically pure compound to form a diastereomeric mixture, followed by separation of the individual diastereomers by standard methods, such as fractional crystallization or chromatography. The coupling reaction is often the formation of salts using an enantiomerically pure acid or base. The diastereomeric derivatives may then be converted to the pure enantiomers by cleavage of the added chiral residue. The racemic mixture of the compounds can also be separated directly by chromatographic methods using chiral stationary phases, which methods are well known in the art. Alternatively, any enantiomer of a compound may be obtained by stereoselective synthesis using optically pure starting materials or reagents of known configuration by methods well known in the art.

[0100] The compounds may possess tautomeric forms, and tautomers also constitute embodiments of the disclosure.

[0101] The present disclosure also includes isotopically-labeled compounds, which are identical to those compounds disclosed herein (including those compounds specifically exemplified herein), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the disclosure are hydrogen, carbon,CCF-43944.601

[0102] nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as, but not limited to2H,3H,13C,14C,15N,180,31P,35S,18F, and36C1, respectively. Substitution with heavier isotopes such as deuterium, i.e.2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be particularly useful in some circumstances. The compound may incorporate positron-emitting isotopes for medical imaging and positron-emitting tomography (PET) studies for determining the distribution of receptors. Suitable positron-emitting isotopes that can be incorporated into the compounds arenC,13N,15O, and18F. Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described herein using an appropriate isotopically-labeled reagent in place of a non-isotopically-labeled reagent.

[0103] The compounds disclosed herein can be synthesized by a variety of methods, including those illustrated in the Examples, in which I3P is activated to form the corresponding N-acylbenzotriazole, which is then reacted with an oligosaccharide to form an ester linkage.

[0104] Compounds and intermediates may be isolated and purified by methods well-known to those skilled in the art of organic synthesis. Examples of conventional methods for isolating and purifying compounds can include, but are not limited to, chromatography on solid supports such as silica gel, alumina, or silica derivatized with alkylsilane groups, by recrystallization at high or low temperature with an optional pretreatment with activated carbon, thin-layer chromatography, distillation at various pressures, sublimation under vacuum, and trituration, as described for instance in “Vogel’s Textbook of Practical Organic Chemistry,” 5th edition (1989), by Furniss, Hannaford. Smith, and Tatchell, pub. Longman Scientific & Technical, Essex CM202JE, England.

[0105] Reaction conditions and reaction times for each individual step can vary depending on the particular reactants employed and substituents present in the reactants used. Reactions can be worked up in a conventional manner, e.g., by eliminating the solvent from the residue and further purified according to methodologies generally known in the art such as. but not limited to, crystallization, distillation, extraction, trituration and chromatography. Unless otherwise described, the starting materials and reagents are either commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature.CCF-43944.601

[0106] Standard experimentation, including appropriate manipulation of the reaction conditions, reagents and sequence of the synthetic route, protection of any chemical functionality that cannot be compatible with the reaction conditions, and deprotection at a suitable point in the reaction sequence of the method are included in the scope of the disclosure. Suitable protecting groups and the methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which can be found in PGM Wuts and TW Greene, in Greene’s book titled Protective Groups in Organic Synthesis (4thed.), John Wiley & Sons, NY (2006).

[0107] When an optically active form of a disclosed compound is required, it can be obtained by carrying out one of the procedures described herein using an optically active starting material (prepared, for example, by asymmetric induction of a suitable reaction step), or by resolution of a mixture of the stereoisomers of the compound or intermediates using a standard procedure (such as chromatographic separation, recrystallization or enzymatic resolution).

[0108] Similarly, when a pure geometric isomer of a compound is required, it can be obtained by carrying out one of the procedures described herein using a pure geometric isomer as a starting material, or by resolution of a mixture of the geometric isomers of the compound or intermediates using a standard procedure such as chromatographic separation.

[0109] The synthetic schemes and specific examples as described are illustrative and are not to be read as limiting the scope of the disclosure or the claims. Alternatives, modifications, and equivalents of the synthetic methods and specific examples are contemplated.

[0110] The disclosed compounds may exist as pharmaceutically acceptable salts. The term “pharmaceutically acceptable salt” refers to salts or zwitterions of the compounds which are water or oil-soluble or dispersible, suitable for treatment of disorders without undue toxicity, irritation, or allergic response, commensurate with a reasonable benefit / risk ratio and effective for their intended use. The salts may be prepared during the final isolation and purification of the compounds or separately by reacting an amino group of the compounds with a suitable acid. For example, a compound may be dissolved in a suitable solvent, such as but not limited to methanol and water, and treated with at least one equivalent of an acid, such as hydrochloric acid. The resulting salt may precipitate out and be isolated by filtration and dried under reduced pressure. Alternatively, the solvent and excess acid may be removed under reduced pressure to provide a salt. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate,CCF-43944.601

[0111] benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3 -phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, para-toluenesulfonate, undecanoate, hydrochloric, hydrobromic, sulfuric, phosphoric and the like. The amino groups of the compounds may also be quatemized with alkyl chlorides, bromides and iodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, and the like. In one embodiment, the compound is in the form of a halide salt, such as a chloride or bromide salt.

[0112] Basic addition salts may be prepared during the final isolation and purification of the disclosed compounds by reaction of a carboxyl group with a suitable base such as the hydroxide, carbonate, or bicarbonate of a metal cation such as lithium, sodium, potassium, calcium, magnesium, or aluminum, or an organic primary, secondary, or tertiary amine. Quaternary amine salts can be prepared, such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, AQV-dimethylaniline, N-methylpiperidine, IV-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1 -ephenamine and N,N’ -dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like.

[0113] Compounds disclosed herein can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure. Certain compounds of the disclosure may also exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0114] Pharmaceutical Compositions

[0115] The disclosed compounds (i.e., compounds of formula (I), or pharmaceutically acceptable salts thereof) may be incorporated into pharmaceutical compositions suitable for administration to a subject. The pharmaceutical compositions may include a therapeutically effective amount of the compound.

[0116] The pharmaceutical compositions may include pharmaceutically acceptable carriers. The term “pharmaceutically acceptable carrier,” as used herein, means a non-toxic, inert solid, semi-CCF-43944.601

[0117] solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as, but not limited to, lactose, glucose and sucrose; starches such as. but not limited to, com starch and potato starch; cellulose and its derivatives such as, but not limited to, sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as, but not limited to, cocoa butter and suppository waxes; oils such as, but not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols; such as propylene glycol; esters such as, but not limited to, ethyl oleate and ethyl laurate; agar; buffering agents such as, but not limited to, magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as. but not limited to, sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0118] Thus, the compounds and their pharmaceutically acceptable salts may be formulated for administration by, for example, solid dosing, eye drop, in a topical oil-based formulation, injection, inhalation (either through the mouth or the nose), implants, or oral, buccal, parenteral, or rectal administration. Techniques and formulations may generally be found in “Remington’s Pharmaceutical Sciences,” (Meade Publishing Co., Easton, Pa.). Therapeutic compositions must typically be sterile and stable under the conditions of manufacture and storage.

[0119] The route by which the disclosed compounds are administered and the form of the composition will dictate the type of carrier to be used. The composition may be in a variety of forms, suitable, for example, for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implants, or parenteral) or topical administration (e.g., dermal, pulmonary, nasal, aural, ocular, liposome delivery systems, or iontophoresis).

[0120] Carriers for systemic administration typically include at least one of diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, combinations thereof, and others. All carriers are optional in the compositions.

[0121] Suitable diluents include sugars such as glucose, lactose, dextrose, and sucrose; diols such as propylene glycol; calcium carbonate; sodium carbonate; sugar alcohols, such as glycerin;CCF-43944.601

[0122] mannitol; and sorbitol. The amount of diluent(s) in a systemic or topical composition is typically about 50 to about 90% by weight of the composition.

[0123] Suitable lubricants include silica, talc, stearic acid and its magnesium salts and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycol and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, com oil and oil of theobroma. The amount of lubricant(s) in a systemic or topical composition is typically about 5 to about 10% by weight of the composition.

[0124] Suitable binders include polyvinyl pyrrolidone; magnesium aluminum silicate; starches such as corn starch and potato starch; gelatin; tragacanth; and cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, methylcellulose, microcrystalline cellulose, and sodium carboxymethylcellulose. The amount of binder(s) in a systemic composition is typically about 5 to about 50% by weight of the composition.

[0125] Suitable disintegrants include agar, alginic acid and the sodium salt thereof, effervescent mixtures, croscarmellose, crospovidone. sodium carboxymethyl starch, sodium starch glycolate, clays, and ion exchange resins. The amount of disintegrant(s) in a systemic or topical composition is typically about 0.1 to about 10% by weight of the composition.

[0126] Suitable colorants include a colorant such as an FD&C dye. When used, the amount of colorant in a systemic or topical composition is typically about 0.005 to about 0.1% by weight of the composition.

[0127] Suitable flavors include menthol, peppermint, and fruit flavors. The amount of flavor(s), when used, in a systemic or topical composition is typically about 0.1 to about 1.0%.

[0128] Suitable sweeteners include aspartame and saccharin. The amount of sweetener(s), when used, in a systemic or topical composition is typically about 0.001 to about 1% by weight of the composition.

[0129] Suitable antioxidants include butylated hydroxyanisole (“BHA”), butylated hydroxytoluene (“BHT”), and vitamin E. The amount of antioxidant(s) in a systemic or topical composition is typically about 0.1 to about 5% by weight of the composition.

[0130] Suitable preservatives include benzalkonium chloride, methyl paraben, and sodium benzoate. The amount of preservative(s) in a systemic or topical composition is typically about 0.01 to about 5% by weight of the composition.CCF-43944.601

[0131] Suitable glidants include silicon dioxide. The amount of glidant(s) in a systemic or topical composition is typically about 1 to about 5% by weight of the composition.

[0132] Suitable solvents include water, isotonic saline, ethyl oleate, glycerin, hydroxylated castor oils, alcohols such as ethanol, and phosphate buffer solutions. The amount of solvent(s) in a systemic or topical composition is typically from about 0 to about 100% by weight of the composition.

[0133] Suitable suspending agents include AVICEL RC-591 (from FMC Corporation of Philadelphia, PA) and sodium alginate. The amount of suspending agent(s) in a systemic or topical composition is typically about 1 to about 8% by weight of the composition.

[0134] Suitable surfactants include lecithin, Polysorbate 80, and sodium lauryl sulfate, and the TWEENS from Atlas Powder Company of Wilmington, Delaware. Suitable surfactants include those disclosed in the C.T.F.A. Cosmetic Ingredient Handbook, 1992, pp.587-592; Remington’s Pharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon’s Volume 1, Emulsifiers & Detergents, 1994, North American Edition, pp. 236-239. The amount of surfactant(s) in the systemic or topical composition is typically about 0.1% to about 5% by weight of the composition.

[0135] Although the amounts of components in the systemic compositions may vary depending on the type of systemic composition prepared, in general, systemic compositions include 0.01% to 50% by weight of an active compound and 50% to 99.99% by weight of one or more carriers. Compositions for parenteral administration typically include 0.1% to 10% by weight of actives and 90% to 99.9% by weight of a carrier including a diluent and a solvent.

[0136] Compositions for oral administration can have various dosage forms. For example, solid forms include tablets, capsules, granules, and bulk powders. These oral dosage forms include a safe and effective amount, usually at least about 5% by weight, and more particularly from about 25% to about 50% by weight of actives. The oral dosage compositions include about 50% to about 95% by weight of carriers, and more particularly, from about 50% to about 75% by weight.

[0137] Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated, or multiple-compressed. Tablets typically include an active component, and a carrier comprising ingredients selected from diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, glidants, and combinations thereof. Specific diluents include calcium carbonate,CCF-43944.601

[0138] sodium carbonate, mannitol, lactose and cellulose. Specific binders include starch, gelatin, and sucrose. Specific disintegrants include alginic acid and croscarmellose. Specific lubricants include magnesium stearate, stearic acid, and talc. Specific colorants are the FD&C dyes, which can be added for appearance. Chewable tablets preferably contain sweeteners such as aspartame and saccharin, or flavors such as menthol, peppermint, fruit flavors, or a combination thereof.

[0139] Capsules (including implants, time release and sustained release formulations) typically include an active compound (e.g., a compound of formula (I)) and a carrier including one or more diluents disclosed above in a capsule comprising gelatin. Granules typically comprise a disclosed compound, and preferably glidants such as silicon dioxide to improve flow characteristics. Implants can be of the biodegradable or the non-biodegradable type.

[0140] The selection of ingredients in the carrier for oral compositions depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of this disclosure.

[0141] Solid compositions may be coated by conventional methods, typically with pH or timedependent coatings, such that a disclosed compound is released in the gastrointestinal tract in the vicinity of the desired application, or at various points and times to extend the desired action. The coatings typically include one or more components selected from the group consisting of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, EUDRAGIT® coatings (available from Evonik Industries of Essen, Germany), waxes and shellac.

[0142] Compositions for oral administration can have liquid forms. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted from non-effervescent granules, suspensions reconstituted from non-effervescent granules, effervescent preparations reconstituted from effervescent granules, elixirs, tinctures, syrups, and the like. Liquid orally administered compositions typically include a disclosed compound and a carrier, namely, a carrier selected from diluents, colorants, flavors, sweeteners, preservatives, solvents, suspending agents, and surfactants. Peroral liquid compositions preferably include one or more ingredients selected from colorants, flavors, and sweeteners.

[0143] Other compositions useful for attaining systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically include one or more of soluble filler substances such as diluents including sucrose, sorbitol and mannitol; andCCF-43944.601

[0144] binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose, and hydroxypropyl methylcellulose. Such compositions may further include lubricants, colorants, flavors, sweeteners, antioxidants, and glidants.

[0145] Methods of Use

[0146] As discussed above, studies have suggested a potential role for IPA in treating a variety of disorders. As such, the compounds disclosed herein (i.e., compounds of formula (I) and pharmaceutically acceptable salts thereof), and pharmaceutical compositions disclosed herein (i.e., pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof) can be used in methods of treating such disorders. As the compounds are disfavored to undergo systemic absorption upon oral administration, and thus have delayed-release properties, the compounds can achieve colon-targeted delivery and represent a promising method to increase and sustain free IPA levels in patients, compared to direct administration of IPA alone.

[0147] Accordingly, disclosed herein is a method of treating a disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (1), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof. Also disclosed herein is a method of increasing levels of indole-3-propionic acid in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof. Also disclosed herein is a method of promoting health, longevity, vitality, vigor, and / or wellness in a subject, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof. Also disclosed herein is a providing radioprotective effects to a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising compound of formula (I), or a pharmaceutically acceptable salt thereof. For example, in some embodiments, the compound of formula (I), or the pharmaceutically acceptable salt thereof, can provide radioprotective effects to subject undergoing radiation therapy, e.g., for the treatment of cancer.CCF-43944.601

[0148] The methods disclosed herein can be used for treatment of any disorder known to be associated with reduced levels of IPA. For example, in some embodiments, the disorder is a cardiovascular disease. The term “cardiovascular disease” (CVD) is used in the art in reference to conditions affecting the heart, heart valves, and vasculature (such as arteries and veins) of the body and encompasses diseases and conditions including, but not limited to, angina, arrhythmia, arteriosclerosis, atherosclerosis, myocardial infarction, acute coronary syndrome, cardiomyopathy, congestive heart failure, coronary thrombosis, arterial aneurysm (e.g., aortic aneurysm, iliac aneurysm, or femoral aneurysm), aortic dissection, pulmonary embolism, high blood pressure / hypertension (e.g., primary hypertension), hypercholesterolemia / hyperlipidemia, atrial fibrillation, stroke, transient ischemic attack, systolic dysfunction, diastolic dysfunction, myocarditis, atrial tachycardia, ventricular fibrillation, endocarditis, arteriopathy, vasculitis, atherosclerotic plaque, vulnerable plaque, acute coronary syndrome, acute ischemic attack, sudden cardiac death, peripheral vascular disease, coronary artery disease (CAD), carotid artery disease, peripheral artery disease (PAD), cerebrovascular disease, adverse ventricular remodeling, ventricular systolic dysfunction, ventricular diastolic dysfunction, cardiac dysfunction, ventricular arrhythmia, stroke, and the like. In some embodiments, the cardiovascular disease is selected from heart failure, coronary heart disease, cor pulmonale, congenital heart defect, cardiomyopathy, myocardial infarction, abdominal aortic aneurysm, valvular heart disease, arrhythmia, peripheral arterial disease, cerebrovascular accident, atherosclerosis, and rheumatic heart disease. In some embodiments, the cardiovascular disease is heart failure, in particular heart failure with preserved ejection fraction (HFpEF).

[0149] For example, abdominal aortic aneurysm (AAA) is a disease that is prevalent in older adults and can cause considerable morbidity and mortality if not treated early. An analysis of patients having AAA revealed that elevated levels of IPA were associated with statistically significant reduced odds for AAA regardless of other variables in the case / control cohort (FIG.

[0150] 2). In addition, Kaplan-Meier survival analysis of baseline IPA showed decreases in the rates of fast-growing aneurysm and the need for surgical repair among subjects. Elevated IPA levels were associated with a statistically significant reduced risk for fast-growing aneurysm and the need for surgical repair (FIG. 3). Accordingly, elevating IPA levels in a patient with AAA through supplementation using a delayed-release form of IPA, may provide a different treatment option.CCF-43944.601

[0151] In some embodiments, the disorder is selected from obesity, diabetes, endotoxin leakage, cancer, inflammation, cell oxidative damage, skin disorders, influenza infection, neurodegenerative diseases, nonalcoholic steatohepatitis. and sepsis. For example, in some embodiments, the disorder is obesity or diabetes. IPA appears to also play a role in protection against obesity and diabetes in obese patients with type 2 diabetes where IPA levels are significantly decreased (Jennis 2018). It has also been reported that the serum concentration of IPA is associated with insulin secretion and that IPA could be a factor in reducing the risk of type 2 diabetes (Tuomainen et al. Nutr Diabetes. 2018, 8:35), and a separate study showed that dietary supplementation of IPA to rats was shown to reduce blood glucose concentration (Abildgaard 2018). In some embodiments, the disorder is influenza infection (see, e.g., Heumela 2024). In some embodiments, the disorder is a neurodegenerative disease, such as Alzheimer’s disease or Parkinson’s disease (see, e.g., Konopelski 2022). In some embodiments, the disorder is sepsis (see, e.g., Fang 2022; Huang 2022; Wang 2023). In some embodiments, the disorder is inflammation or cell oxidative damage (see, e.g., Wikoff 2009; Chyan 1999; de Mello 2017). In some embodiments, the disorder is colitis (Lee 2021). In some embodiments, the disorder is a skin disorder (see, e.g., WO 2020 / 185913 Al).

[0152] In some embodiments, the disorder is cancer, such as triple-negative breast cancer, colon cancer, glioblastoma, ovarian cancer, or leukemia. As also discussed above, a recent study in cells and animals showed that IPA displays numerous anti-cancer activities. For example, as discussed above, IPA has been reported to foster cytostatic properties in breast cancer (Sari 2020) and other cancers (e.g. ovarian, triple negative breast cancer, colon cancer). The putative mechanism-of-action was attributed to the activation of both the aryl hydrocarbon receptor (AHR) and the pregnane X receptor (PXR). Activation of these receptors is inversely related to cancer cell proliferation levels and, in humans, a higher expression of AHR and PXR showed improved survival in breast cancer patients. It was also shown that supplementing mice with IPA reduced metastasis formation. Also, in women with newly diagnosed breast cancer, fecal DNA studies showed that the microbiome’s capacity to make IPA was reduced, suggesting a potential role for IPA in human cancers. Additional evidence for the consideration of IPA having an anticancer role is that AHR has already been linked as a tumor suppressor gene in multiple human cancers (Zudaire et al. J. Clin. Invest. 2008, 118(2), 640-650). Thus, the increasing levelsCCF-43944.601

[0153] of TPA could serve as an important additional treatment option for patients with different types of cancer.

[0154] In some embodiments, the disorder is intestinal endotoxin leakage resulting in steatohepatitis, such as metabolic dysfunction-associated steatohepatitis (MASH). MASH is inflammation of the liver caused by excess fat cells due to increased lipolysis, hepatic lipogenesis, and eating high-calorie diets. Chronic inflammation causes progressive liver damage. MASH is often associated with factors of metabolic syndromes such as obesity and type II diabetes. In some embodiments, the disorder is nonalcoholic steatohepatitis (NASH).

[0155] A compound or composition can be administered by any suitable route. The compound or composition is, in various aspects of the disclosure, delivered to an individual parenterally (for example intravenously, intraperitoneally, intrapulmonary, subcutaneously or intramuscularly), intrathecally, topically, transdermally, rectally, orally, sublingually, nasally or by inhalation. In various embodiments, a compound or a composition is administered to the gastrointestinal tract, such as by ingestion. Although the compounds are expected to have delayed-release properties in and of themselves for reasons discussed above, release can be further sustained or delayed by incorporation into suitable formulations. For example, sustained release formulations may also be employed to achieve a controlled release of the compound when in contact with body fluids in the gastrointestinal tract. Sustained release formulations are known in the art, and typically include a polymer matrix of a biological degradable polymer, a water-soluble polymer, or a mixture of both, optionally with suitable surfactants.

[0156] In some embodiments, the compounds described herein may be provided in a delayed release composition and are optionally released in a specific region of the digestive tract of an individual. For example, the composition may be provided such that the compounds are released from an orally dosed composition in the distal portion of the digestive tract such as the ileum or the colon. In certain embodiments, the delayed release composition releases the compounds at a specific pH, or at a range of pH for targeted delivery within the digestive tract of an individual. The compounds may be released, for example, between pH 6.0 and pH 9.0. between pH 6.5 and pH 8.0, between pH 6.5 and pH 7.5, between pH 7.0 and pH 7.5, or between pH 7.0 and pH 8.0.

[0157] It will be appreciated that appropriate dosages of the compounds, and compositions comprising the compounds, can vary from patient to patient. Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleteriousCCF-43944.601

[0158] side effects of the treatments of the present disclosure. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, and / or materials used in combination, and the age, sex, weight, condition, general health, and prior medical history of the patient. The amount of compound and route of administration will ultimately be at the discretion of the physician, although generally the dosage will be to achieve local concentrations at the site of action which achieve the desired effect without causing substantial harmful or deleterious side-effects.

[0159] Method of treatment described herein may comprise administering a second agent to an individual. The term “second agent,” as used herein, serves to distinguish the agent from the compound of formula (I) (or pharmaceutically acceptable salt thereof) and is not meant to limit the number of additional agents used in a method or denote an order of administration. One or more second agents are optionally incorporated in a composition with the compound of formula (I) (or pharmaceutically acceptable salt thereof), administered concurrently but in separate dosage forms, or administered separately in time.

[0160] Exemplary second agents include, but are not limited to, antimicrobials (such as antibiotics that kill bacteria in the gut); agents that improve intestinal motility (such as fiber or psyllium); agents that improve one or more aspects of cardiovascular health, such as agents that normalize blood pressure, decrease vascular inflammation, reduce platelet activation, normalize lipid abnormalities; a nicotinamide adenine dinucleotide (NAD)-boosting compound; or an a-amino-P-carboxymuconate-s-semialdehyde decarboxylase (ACMSD) inhibitor. In various embodiments, the second agent is selected from the group consisting of an omega-3 oil, salicylic acid (aspirin), dimethylbutanol, garlic oil, garlic extract, olive oil, krill oil, coenzyme Q10, a probiotic, a prebiotic, a dietary fiber, psyllium husk, bismuth salts, phytosterols, grape seed oil, green tea extract, vitamin D, an antioxidant (such as vitamin C and vitamin E), turmeric, curcumin, resveratrol, activated charcoal, or copper chlorophyllin. Additional exemplary second agents are described in US 2017 / 0151208, US 2017 / 0151250, US 2017 / 0152222, or US 2018 / 0000754, which are incorporated here by reference.

[0161] In some embodiments, the second agent is an NAD-boosting compound, such as a supplement. Numerous studies have reported that elevations in intracellular NAD+ lead to antiaging benefits. Exemplary NAD-boosting compounds include, but are not limited to, NAD,CCF-43944.601

[0162] nicotinic acid, nicotinamide, nicotinamide riboside, N-methyl-nicotinamide, niacin, nicotinamide mononucleotide, vitamin B3 or forms thereof, and resveratrol. The NAD-boosting compound may be administered in the form of a supplement.

[0163] In some embodiments, the second agent is an ACMSD inhibitor. This class of agents has potential anti-cancer properties. ACMSD is a key regulator of tryptophan flux into either the tricarboxylic acid (TCA) cycle for use as fuel, or into the NAD pool to boost NAD levels. While dietary tryptophan is primarily used as a fuel into the TCA cycle, with ACMSD inhibition, it is diverted to NAD generation. This in turn can lead to excess NAD and formation of 4PY. IPA inhibits expression of the rate-limiting enzyme in the synthesis of 4PY, which is nicotinamide N-methyltransferase (NNMT) (Wang 2024).

[0164] Methods disclosed herein may further comprise administration of one or more cardiovascular disease therapies. Examples of therapies include, but are not limited to, statins (e.g., Lipitor™ (atorvastatin), Pravachol™ (pravastatin), Zocor™ (simvastatin), Mevacor™ (lovastatin), and Lescol™ (fluvastatin)) or other agents that interfere with the activity of HMGCoA reductase, nicotinic acid (niacin, which lowers LDL cholesterol levels), fibrates (which lower blood triglyceride levels and include, for example Bezafibrate (such as Bezalip®), Ciprofibrate (such as Modalim®), Clofibrate, Gemfibrozil (such as Lopid®) and Fenofibrate (such as TriCor®)), bile acid resins (such as Cholestyramine, Colestipol (Colestid), and Cholsevelam (Welchol)), cholesterol absorption inhibitors (such as Ezetimibe (Zetia®, Ezetrol®, Ezemibe®)), phytosterols such as sitosterol (Take Control (Lipton)), sitostanol (Benechol), or stigmastanol), alginates and pectins, lecithin, and nutraceuticals (such as extract of green tea and other extracts that include polyphenols, particularly epigallocatechin gallate (EGCG), Cholest-Arrest™ (500 mg garlic and 200 mg lecithin). Cholestaway™ (700 mg Calcium carbonate, 170 mg magnesium oxidem 50 pg chromium picolinate), Cholest-Off™ (900 mg of plant sterols / stanols), Guggul Bolic (750 mg gugulipid (Commiphora mukul gum resin), and Kyolic® (600 mg aged garlic extract and 380 mg lecithin)).

[0165] In methods of treating cancer, in some embodiments, a compound or composition disclosed herein can be administered in combination with other therapeutic treatment modalities, including surgery, chemotherapy, radiation therapy, hormone therapy, immunotherapy, cryotherapy, and thermotherapy. Such combination therapies may advantageously utilize lower dosages of the administered agent and / or other chemotherapeutic agent, thus avoiding possibleCCF-43944.601

[0166] toxicities or complications associated with the various therapies. In some embodiments, the compound described herein is administered with at least one additional therapeutic agent, such as a chemotherapeutic agent. In certain embodiments, the compound described herein is administered in combination with one or more additional chemotherapeutic agents. The chemotherapeutic agent may be a chemotherapeutic agent identified on the “A to Z List of Cancer Drugs” published by the National Cancer Institute.

[0167] Kits

[0168] For use in the methods described herein, kits and articles of manufacture are also provided, which include a compound or pharmaceutical composition described herein (e.g., a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof). In some embodiments, such kits comprise a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers are formed from a variety of materials such as glass or plastic.

[0169] The articles of manufacture provided herein contain packaging materials. Packaging materials for use in packaging pharmaceutical products include those found in, e.g., U.S. Patent Nos. 5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment. For example, in some embodiments the container(s) includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, optionally in a composition or in combination with another agent as disclosed herein. The container(s) optionally have a sterile access port (for example the container is an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). Such kits optionally comprisie a compound with an identifying description or label or instructions relating to its use in one or more methods described herein.

[0170] For example, a kit typically includes one or more additional containers, each with one or more of various materials (such as reagents, optionally in concentrated form, and / or devices) desirable from a commercial and user standpoint for use of a compound described herein. Non-CCF-43944.601

[0171] limiting examples of such materials include, but not limited to, buffers, diluents, filters, needles, syringes; carrier, package, container, vial and / or tube labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included. A label is optionally on or associated with the container. For example, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In addition, a label is used to indicate that the contents are to be used for a specific therapeutic application. In addition, the label indicates directions for use of the contents, such as in the methods described herein. In certain embodiments, the pharmaceutical composition is presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. Or, the pack or dispenser device is accompanied by instructions for administration. Or, the pack or dispenser is accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. In some embodiments, compositions containing a compound provided herein formulated in a compatible pharmaceutical earner are prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0172] Examples

[0173] Abbreviations used in the Examples include the following: DMF is dimethylformamide; EtOAc is ethyl acetate; IPA is indole-3-propionic acid; and TLC is thin-layer chromatography.

[0174] Example 1: Syntheses of Indole-3-Propionic Acid Conjugate Esters l-( lH-benzo[d][ 1,2,3 ]triazol-l-yl)-3-( 1 H-indol-3-yl)propan-l -one

[0175]

[0176] CCF-43944.601

[0177] Prepared according to a previously-disclosed procedure (Katritzky el al. Synthesis 2003; 18:2795-2798). In a 2000 mL flask, thionyl chloride (14.2 mL, 195 mmol) was added to a stirred solution of IH-benzotriazole (81.00 g, 679.9 mmol) in dichloromethane (750 mL) at room temperature over a period of 2 minutes. The resulting yellow solution stirred for 30 min and then additional dichloromethane added (250 mL) and the solution briefly cooled in an ice-bath for 10 min. Solid 3-indolepropionic acid (36.76 g, 194.3 mmol) added in 1 portion whereupon a heavy precipitation occurred after 1 min. The flask was swirled to break up the precipitate. The resulting mixture stirred at room temperature for 1 h and then suction filtered through filter paper (grade 1). The filtered solid was rinsed with dichloromethane (2 x 75 mL). Filtrate collected and washed with a mixture of saturated aqueous sodium bicarbonate (200 mL) and aqueous NaOH (2 M, 50 mL). The aqueous layer remained basic (pH~8-9) by pH paper. Dichloromethane layer dried over sodium sulfate, decanted, and concentrated using rotary evaporation to give a tan solid. The solid product mixture was washed (flask swirled for ~30 s) with 40% EtOAc-heptane (100 mL) and then the solid allowed to settle for 5 min and the wash solution was decanted off to remove excess benzotriazole. This washing process was repeated twice (2 x 100 mL). A final wash of 40% EtOAc-heptane (100 mL) followed by filtration through filter paper (grade 4) gave l-(lH-benzo[d][l,2,3]triazol-l-yl)-3-(lH-indol-3-yl)propan-l-one as a tan-white solid which was air dried for 2 d, 46.12 g, 82% yield.

[0178] Alpha-cyclodextrin-IPA conjugate

[0179]

[0180] Representative structure

[0181] Product is a distribution of substitutions

[0182] In a 250 mL 2-neck reaction flask, a mixture of l-(lH-benzo[d][l,2,3]triazoLl-yl)-3-(lH-indol-3-yl)propan-l-one (4.9431 g, 17.026 mmol) and alpha-cyclodextrin (4.1472 g, 4.2630CCF-43944.601

[0183] mmol) was dissolved in dimethylformamide (50 mL) and pyridine (10 mL). The resulting solution stirred at 70 °C (oil bath temperature) overnight. TLC (35% ethyl acetate-heptane) showed complete consumption of the indole starting material. Reaction mixture transferred to a single-neck 250 ml flask and partially concentrated using rotary evaporation (0 bar at 50 °C).

[0184] The remaining solution was concentrated using high vacuum to remove most of the DMF. The resulting crude product mixture was triturated with ethyl acetate (150 mL) to remove the liberated benzotriazole. The large solid chunks were broken up using a spatula and the mixture suction filtered through grade 1 filter paper. The recovered solid was rinsed with ethyl acetate (100 mL). The resulting tan solid collected and dried under high vacuum to give the product, 6.0854 g, 86% yield. Mass spec showed desired mass peaks at [M + Na] = 1337 (2 substitutions), [M + Na] = 1508 (3 substitutions), [M + Na] = 1680 (4 substitutions). Measured IPA loading after hydrolysis using aqueous sodium hydroxide, neutralizing, and then mass spectrometry analysis: 31.2 wt% IPA

[0185] Beta-cyclodextrin-IPA conjugate

[0186]

[0187] In a 250 mL 2-neck reaction flask, a mixture of l-(lH-benzo[d][l,2,3]triazol-l-yl)-3-(lH-indol-3-yl)propan-l-one (4.1782 g, 14.391 mmol) and beta-cyclodextrin (3.6365 g, 3.2040 mmol) was dissolved in dimethylformamide (50 mL) and pyridine (10 mL). The resulting solution stirred at 70 °C (oil bath temperature) overnight. TLC (35% ethyl acetate-heptane) showed the presence of the indole starting material. Additional beta-cyclodextrin added (1.3082 g, 1.1526 mmol) and the resulting solution stirred at 75 °C overnight. TLC showed only a very small amount of starting material. Reaction mixture cooled to room temperature and transferredCCF-43944.601

[0188] to a single-neck 250 ml flask and partially concentrated using rotary evaporation (0 bar at 50 °C). The remaining solution was concentrated using high vacuum to remove most of the DMF. The resulting crude product mixture was triturated with ethyl acetate (150 mL) to remove the

[0189] liberated benzotriazole. The large solid chunks were broken up using a spatula and the mixture suction filtered through grade 1 filter paper. The recovered solid was rinsed with ethyl acetate (100 mL). The resulting white solid collected and dried under high vacuum to give 6.9776 g of product. Mass spec showed a significant peak at 1157 (consistent with beta-cyclodextrin [M++ Na]). Solid product added to water (75 mL) and stirred for 45 min at room temperature to dissolve the free beta-cyclodextrin. Mixture suction filtered through grade 1 filter paper and solid dried in ambient air overnight and then high vacuum for 4 h to give a white solid, 5.3651 g, 72% yield. Mass spec showed desired mass peaks at [M + Na] = 1328.5 (1 substitution), [M + Na] = 1499.6 (2 substitutions), [M + Na] = 1671.6 (3 substitutions), [M + Na] = 1842.6 (4 substitutions). Measured IPA loading after hydrolysis using aqueous sodium hydroxide, neutralizing, and then mass spectrometry analysis: 35.9 wt% IPA

[0190] Gamma-cyclodextrin-IPA conjugate

[0191]

[0192] In a 500 mL round bottom flask, a mixture of l-(lH-benzo[d][l,2,3]triazol-l-yl)-3-(lH-indol-3-yl)propan-l-one-l (40.02 g, 137.8 mmol) and gamma-cyclodextrin (35.91 g, 27.68 mmol) was diluted with dimethylformamide (160 mL) and pyridine (40 mL). The resulting heterogeneous mixture stirred at 75 °C (oil bath temperature) overnight (22 h). The next day, the reaction mixture was homogeneous. TLC (35% ethyl acetate-heptane) did not show the indole starting material. Reaction mixture cooled to room temperature and concentrated using rotaryCCF-43944.601

[0193] evaporation (0 bar at 60 °C). The remaining solution was concentrated using high vacuum for 4 h to remove most of the DMF to give a thick syrup. The resulting crude product thick syrup was triturated with 85% ethyl acetate- hexane (300 mL) to remove the liberated benzotriazole and any remaining starting material if present. A spatula was used to mix the syrup in the ethyl acetatehexane wash and a solid formed after mixing for 5 min. The resulting mixture sat for 45 min. The ethyl acetate-hexane wash was decanted off and discarded. Ethyl acetate (300 mL) was then added to the solid and the washing (agitating with a spatula) / decanting step repeated. Additional ethyl acetate (300 mL) added and the mixture agitated with a spatula for 10 min and then suction filtered through grade 1 filter paper. The recovered solid was rinsed with ethyl acetate (100 mL). The resulting light tan solid air dried for 2 h. The resulting light tan solid collected and dried under high vacuum for 22 h to give the product. 61.31 g, 103% yield. Mass spec showed desired mass peaks at [M + Na] = 1490.6 (1 substitution), [M + Na] = 1661.5 (2 substitutions), [M + Na] = 1832.8 (3 substitutions). Measured IPA loading after hydrolysis using aqueous sodium hydroxide, neutralizing, and then mass spectrometry analysis: 36.6 wt% IPA.

[0194] Inulin-IPA conjugate

[0195]

[0196] Representative structure.

[0197] Product is a distribution of substitutions In a 500 mL flask was added a mixture of l-(lH-benzo[d][l,2,3]triazol-l-yl)-3-(lH-indol-3-yl)propan-l-one (42.90 g, 147.8 mmol) and inulin (41.87 g). Dimethylformamide (160 mL) and pyridine (50 mL) were then added to the flask. The resulting mixture stirred at 75 °C (oil bath temperature) whereupon a clear solution formed after ~20 min. The solution was stirred for 24 h at 75 °C. TLC (35% ethyl acetate-heptane) of a sample directly from the reaction mixture (very concentrated) showed a very small amount of the indole starting material. Reaction mixture cooled to room temperature and mostly concentrated using rotary evaporation (0 bar atCCF-43944.601

[0198] 60 °C). Water (50 mL) was added and the mixture re-subjected to the same rotary evaporation conditions to help azeotrope additional DMF. The remaining syrup was concentrated using high vacuum for 4 h to give a thick syrup. The resulting crude product mixture was triturated with 90% ethyl acetate-hexanes (300 mL) to remove the liberated benzotriazole. A spatula was used to mix the crude product in the ethyl acetate wash. The resulting large solid chunks were broken up using a spatula and the mixture sat for 45 min. The ethyl acetate-hexanes wash was decanted off and discarded. Ethyl acetate (300 mL) added and the washing (agitating with a spatula) / decanting step repeated. Additional ethyl acetate (300 mL) added and the mixture agitated with a spatula for 10 min and then suction filtered through grade 1 filter paper. The recovered solid was rinsed with ethyl acetate (100 mL). The resulting light tan solid collected and air dried for 3 d and then dried under high vacuum for 24 h to give the product, 58.47 g, 87% yield. Measured IPA loading after glycoside cleavage using aqueous HC1 and ester hydrolysis using aqueous sodium hydroxide, neutralizing, and then mass spectrometry analysis: 37.3 wt% IPA.

[0199] Example 2: Assay Data

[0200] Pharmacokinetics and Fecal IPA Recovery

[0201] Suspensions of IPA conjugates were prepared as follows, and given as oral gavage to mice. The IPA-y-cyclodextrin (BP-y-cyclodextrin) conjugate was prepared as a fine aqueous suspension containing 20% dimethyl sulfoxide and 2% sucrose. The IPA-inulin conjugate prepared as a fine aqueous suspension containing 20% sucrose and 0.06% sodium dodecyl sulfate. Free IPA was gavaged as an aqueous suspension containing 20% DMSO and 0.8% aq sodium carboxymethylcellulose.

[0202] C57bl / 6 female mice (11 mice tested per IPA form, 20-30 g body weight) on a normal chow diet with free access to food and water were given a PO gavage of 0.2 mL containing 100 mg / kg (IPA mass equivalent) IPA-conjugate or 2 mg / kg free IPA. Mice were singly housed in cages fitted with a wire bottom and absorbent material starting at the zero hour dosing time point. Blood and urine were collected at the timepoints indicated (0-24 hours) post-gavage. Feces was pooled from the bottom of the cage at the end of the 24 hours for mass spectrometry analysis of total IPA content (following base hydrolysis with aqueous sodium hydroxide (2.5 M) at 60 °C for 2 hours to hydrolyze the ester, followed by the addition formic acid to until a neutral pH wasCCF-43944.601

[0203] achieved) using established stable isotope dilution LC / MS / MS methods. Plasma, urine, and fecal IPA levels were measured by stable-isotope dilution mass spectrometry using high-performance liquid chromatography in tandem with electrospray ionization mass spectrometry (Shimadzu 8060 mass spectrometer) as previously described (Nemet et al. Eur. Heart J. 2023, 44(32):3085-3096).

[0204] Data are shown in FIGS. 4A-4B, and demonstrate that IPA ester conjugates possess a slow-release profile in mice, and the ability to recover free IPA in mouse feces shows that IPA reaches the colon. A pharmacokinetic profiling study showed that oral administration of both the gamma-cyclodextrin-IPA conjugate and the inulin-IPA conjugate in mice had significantly increased systemic exposure compared to free IPA dosing (FIG. 4A). There is delayed I3P release of the ester conjugates (Inulin and gamma-cyclodextrin forms) in mice compared to free I3P, indicated by the shift to later time points for elevated plasma I3P levels. The table of Cmax (maximum plasma concentration) shown are derived from plasma I3P determined in the monitored period. Thus, oral administration of these IPA conjugates would have a much smaller peak-to-trough concentration fluctuation than the free IPA dosing and would likely sustain IPA levels for a longer period of time compared to the free IPA dosing, ree IPA levels gradually increased over the course of 14 days indicating an accumulation effect consistent with the longer exposure of free IPA via the IPA conjugate. The IPA release from oral administration of the gamma-cyclodextrin and inulin conjugates is also slow enough such that much higher amounts of free IPA can be recovered in mouse feces (FIG. 4B). Conversely, oral free IPA administration resulted in very little recovery in mouse feces.

[0205] Ad libitum feeding study

[0206] Swiss Webster female mice (Taconic) were placed (ad libitum) on either a no choline-added chow (Teklad) diet, or high fat (HF) diet (D12451i from Research Diets). Blood samples were collected at the indicated days (baseline vs up to 14 d after diet change) to determine the impact of I3P supplementation on plasma I3P levels. On day 0, mice were placed on I3P supplemented chow or HF diets (0.189% w / w I3P-inulin; 0.946 g Inulin-IPA ester mixed in 500 g of diet; 100 mg / kg / day I3P equivalent). Blood was collected twice per day during the light cycle (morning and afternoon).CCF-43944.601

[0207] Plasma T3P levels were measured by established stable-isotope dilution mass spectrometry methods using high-performance liquid chromatography in tandem with electrospray ionization mass spectrometry (Shimadzu 8060 mass spectrometer).

[0208] Data are shown in FIG. 5. Free IPA levels gradually increased over the course of 14 days indicating an accumulation effect consistent with the longer exposure of free IPA via the IPA conjugate

[0209] Thus, IPA ester conjugates described in this disclosure achieve colon-targeted delivery and represent a new method to increase and sustain free IPA levels in patients in need.

Claims

CCF-43944.601CLAIMS1. A compound of formula (I)or a pharmaceutically acceptable salt thereof, wherein:A is an oligosaccharide or polysaccharide; andn is at least 1.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A is a cyclodextrin selected from a-cyclodextrin, p-cyclodextrin, and y-cyclodextrin.

3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the cyclodextrin is a-cyclodextrin.

4. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the cyclodextrin is P-cyclodextrin.

5. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the cyclodextrin is y-cyclodextrin.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A is selected from inulin, dextran, and dextrin.

7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein A is inulin.CCF-43944.6018. The compound of any one of claims 1 -7, or a pharmaceutically acceptable salt thereof, wherein the oligosaccharide or polysaccharide is further functionalized with one or more substituents independently selected from the group consisting of alkyl groups, hydroxyalkyl groups, and amino acids.

9. The compound of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein n is 1-100, 1-50, 1-30, or 1-10.

10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) has a calculated polar surface area of greater than 400 and a cLogP of less than zero.

11. A pharmaceutical composition comprising a compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable earner.

12. A method of treating a disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 11.

13. The method of claim 12, wherein the disorder is selected from cardiovascular diseases.

14. The method of claim 13, wherein the cardiovascular disease is selected from heart failure, coronary heart disease, cor pulmonale, congenital heart defect, cardiomyopathy, myocardial infarction, abdominal aortic aneurysm, valvular heart disease, arrhythmia, peripheral arterial disease, cerebrovascular accident, atherosclerosis, and rheumatic heart disease.

15. The method of claim 14, wherein the heart failure is heart failure with preserved ejection fraction (HFpEF).

16. The method of claim 12, wherein the disorder is selected from obesity, diabetes, endotoxin leakage, cancer, inflammation, cell oxidative damage, skin disorders, influenzaCCF-43944.601infection, neurodegenerative diseases, nonalcoholic steatohepatitis, sepsis, aging, frailty, and sarcopenia.

17. The method of claim 16, wherein the disorder is a cancer selected from triple-negative breast cancer, colon cancer, glioblastoma, ovarian cancer, and leukemia.

18. The method of claim 16, wherein the disorder is type II diabetes.

19. The method of claim 18, wherein the disorder is intestinal endotoxin leakage resulting in steatohepatitis.

20. The method of claim 19, wherein the steatohepatitis comprises metabolic dysfunction-associated steatohepatitis.

21. The method of any one of claims 12-20, further comprising administering a second therapeutic agent to the subject.

22. The method of claim 21, wherein the second therapeutic agent is selected from antimicrobials, agents that improve intestinal motility, cardiovascular drugs, a nicotinamide adenine dinucleotide (NAD)-boosting compound; and / or an a-amino-|3-carboxymuconate-s-semialdehyde decarboxylase (ACMSD) inhibitor.

23. The method of claim 21, wherein the second therapeutic agent is selected from Omega 3 oil, salicylic acid, dimethylbutanol, garlic oil, olive oil, krill oil, Co enzyme Q-10, a probiotic, a prebiotic. dietary fiber, psyllium husk, bismuth salts, phytosterols, grape seed oil, green tea extract, vitamin D, antioxidants, turmeric, curcumin, and / or resveratrol.

24. The method of claim 21, wherein the second therapeutic agent is a chemotherapeutic agent.CCF-43944.60125. A method of increasing levels of indole-3-propionic acid in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 11.

26. A method of promoting health, longevity, vitality, vigor, and / or wellness in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 11.

27. A method of providing radioprotective effects to a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 11.

28. The method of claim 27, wherein the subject is undergoing radiation therapy for treatment of cancer.

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