Fatty acid esters of hydroxy fatty acids, synthesis and uses thereof

Abstract

This disclosure concerns a method of synthesizing fatty acid esters of hydroxy fatty acids (FAHFAs), the compounds, derivatives and analogs thereof. This disclosure also concerns the use of FAHFAs to treat a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption, and / or remodel a gut microbiome in a digestive system.

Description

[0001] Fatty Acid Esters of Hydroxy Fatty Acids, Synthesis and Uses Thereof

[0002] Technical Field

[0003] The present disclosure relates, in general terms, to fatty acid esters of hydroxy-fatty acids (FAHFAs), their synthesis and their use in inhibiting lipid absorption.

[0004] Background

[0005] The global prevalence of obesity has reached alarming levels, with 39% of the world's adult population being overweight, as reported by the World Health Organization. This translates to approximately 1.9 billion overweight individuals, of which 650 million or 13% are classified as obese, necessitating more intensive weight control measures. The implications of obesity extend beyond physical appearance, as it is a significant risk factor for various non-communicable diseases, including type 2 diabetes, hypertension, cardiovascular disease, and metabolic dysfunction-associated fatty liver disease. These comorbidities underscore the urgent need for effective anti-obesity therapies to alleviate the associated healthcare burden.

[0006] Effective therapy that promotes weight loss in a safe and relatively non-invasive manner is an enormous and growing market due to the increasing prevalence of obesity worldwide. One notable development in the field of anti-obesity therapy is the emergence of semaglutide, an incretin mimetic commercialized by Novo Nordisk, which received approval from the US Food and Drug Administration for weight management in adults with obesity or are overweight in 2021. The rapid adoption of this drug in the US healthcare system is evidenced by its substantial revenue generation, amounting to approximately US$12.27 billion in the first 9 months of 2023 for Novo Nordisk. Despite the success of semaglutide, the anti-obesity market is projected to expand significantly, potentially reaching US$100 billion by 2030. However, the high cost and potential side effects associated with semaglutide and similar incretin mimetics highlight the need for the development of safer and more affordable anti-obesity drugs. This includes gastrointestinal distress, restricted accessibility and long-term tolerability for many patients.

[0007] There is a growing interest in exploring the potential role of low cost, general use entities that can be used as food additives to lower lipid absorption. Fatty acid esters of hydroxy fatty acids (FAHFAs), such as 9-palmitic acid hydroxy stearic acid (9-PAHSA), havedemonstrated notable anti-diabetic and anti-inflammatory effects. A study investigated the impact of 9-PAHSA on obesity-related inflammation by measuring the levels of TNF-α and IL-1β in adipose tissue macrophages (ATMs) of mice fed with 9-PAHSA for 3 days. The results revealed a decrease in the expression of TNF-α and IL-1β in ATMs of mice fed a high-fat diet, suggesting that 9-PAHSA exerts a mitigating effect on obesity-related inflammation.

[0008] FAHFAs such as 5-PAHSA and 9-PAHSA are commercially available but are associated with a significant cost, conceivably due to complex synthesis methods (Figure 1A). Presently, primary research efforts focused on the synthesis of novel and distinct fatty acid esters of hydroxy fatty acids (FAHFAs) are primarily concentrated on enzymatic and chemical synthesis methods. Notably, current enzymatic synthesis details a bi-enzymatic cascade reaction for the efficient synthesis of FAHFAs using renewable unsaturated fatty acids. In contrast, various chemical synthesis approaches have also been documented (Figure 1C). For example, a regiospecific total synthesis of several branched FAHFAs for LC-MS / MS analysis was proposed. The total synthesis of FAHFAs involved seven steps commencing from terminal alkenes and alkynes. Another method utilized epoxy chloropropane and alkenyl compounds to synthesize enantiomers of ( / ?)-9-palmitic acid hydroxy stearic acid (R-9-PAHSA). While chemical syntheses of FAHFAs offer the advantage of tunability in the hydroxy moiety by altering the chain length of starting substrates, they are often characterized by multi-step procedures that are laborious, intricate, and expensive.

[0009] It would be desirable to overcome or ameliorate at least one of the above-described problems.

[0010] Summary

[0011] The present disclosure provides a method of synthesising a compound of Formula (I), or a salt, solvate, stereoisomer, prodrug or derivative thereof, comprising:

[0012]

[0013] wherein

[0014] L is a linker selected from oxylene, oxyacylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0015] m is an integer selected from 0 to 20;

[0016] n is an integer selected from 0 to 20;i is an integer selected from 0 to 20;

[0017] a) conjugating an alkyl moiety via linker to the hydroxy-alkyl ester of Formula (V) in order to form a compound of Formula (VI);

[0018] OH O O

[0019]

[0020] (V) (VI)

[0021] wherein Ri is optionally substituted alkyl; and

[0022] b) hydrolysing the ester in order to form the compound of Formula (I).

[0023] In some embodiments, step a) comprises reacting the compound of Formula (V) with a compound of Formula (VII) to form a compound of Formula (Via).

[0024] OH O

[0025] (V)

[0026]

[0027] wherein X is halo.

[0028] In some embodiments, step a) comprises oxidising the compound of Formula (V) into a compound of Formula (XIV), reducing the compound of Formula (XIV) in the presence of an amine to form a compound of Formula (VIII), and reacting the compound of Formula (VIII) with a compound of Formula (VII) to form a compound of Formula (Vlb):

[0029]

[0030] (Vlb) wherein R2 is independently selected from H and optionally substituted alkyl; and X is halo.

[0031] In some embodiments, step a) comprises halogenating followed by azidating the compound of Formula (V) to afford compound of Formula (IX), and subjecting the compound of Formula (IX) to a click reaction to form compound of Formula (Vic):Halogenation NaN3> M O-R

[0032] (IX)

[0033]

[0034] (IX) (Vic)

[0035] In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XI) to form a compound of Formula (Vid):

[0036]

[0037] (V) (XI) (Vid) wherein X is halo.

[0038] In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XII) to form a compound of Formula (Vie):

[0039] OH O

[0040]

[0041] (XII) (Vie) wherein X is halo.

[0042] In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XIII) to form a compound of Formula (VIf):

[0043] OH O

[0044]

[0045] (Vlf) wherein X is halo.

[0046] In some embodiments, the method further comprises a step before step a) of reacting a compound of Formula (II) with a compound of Formula (III) in order to form a compound of Formula (IV);OH, COOH ^MgX \pJx^co°H m VJn' 'm

[0047]

[0048] (II) (Hl) (IV) wherein X is halo; and

[0049] protecting the compound of Formula (IV) as a compound of Formula (V);

[0050] OH OH O x^ vxkz^COOH

[0051] VJn

[0052]

[0053] (IV) (V)

[0054] wherein Ri is optionally substituted alkyl.

[0055] In some embodiments, the compound of Formula (I) is a compound of Formula (la-f):

[0056]

[0057] wherein

[0058] i is an integer selected from 3 to 20;

[0059] m is an integer selected from 0 to 15; and

[0060] n is an integer selected from 0 to 15.In some embodiments, the compound of Formula (I) is a compound of Formula (1-1):

[0061]

[0062] In some embodiments, L is oxyacylene;

[0063] I is an integer selected from 0 to 15;

[0064] m is an integer selected from 5 to 10; and

[0065] n is an integer selected from 5 to 10.

[0066] In some embodiments, L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0067] i is an integer selected from 3 to 20;

[0068] m is an integer selected from 0 to 15; and

[0069] n is an integer selected from 0 to 15.

[0070] The present disclosure also relates to a compound of Formula (I), or a salt, solvate, stereoisomer, prodrug, or derivative thereof:

[0071]

[0072] wherein

[0073] L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0074] i is an integer selected from 3 to 20;

[0075] m is an integer selected from 0 to 15; and

[0076] n is an integer selected from 0 to 15.

[0077] In some embodiments, the compound of Formula (I) is an enantiomer as represented by Formula (I- 1):

[0078]

[0079] In some embodiments, i is an integer selected from 10 to 15.

[0080] In some embodiments, m is an integer selected from 10 to 15.In some embodiments, n is an integer selected from 1 to 5.

[0081] The present disclosure also relates to a composition, comprising a compound of Formula (I) or a salt, solvate, stereoisomer, prodrug, or derivative thereof, and optionally an excipient.

[0082] The present disclosure also relates to a method of treating a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, derivative or combination thereof to the subject:

[0083]

[0084] wherein

[0085] when L is oxyacylene;

[0086] i is 15, m is 8, and n is 7; or

[0087] I is 3, m is 10, and n is 5; or

[0088] I is 13, m is 10, and n is 5; or

[0089] I is 15, m is 10, and n is 5; or

[0090] i is 11, m is 10, and n is 5; or

[0091] when L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0092] I is an integer selected from 0 to 20;

[0093] m is an integer selected from 0 to 15; and

[0094] n is an integer selected from 0 to 15.

[0095] The present disclosure also relates to a method of remodeling a gut microbiome in a digestive system of a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, derivative or combination thereof to the subject:

[0096]

[0097] wherein

[0098] when L is oxyacylene;

[0099] i is 15, m is 8, and n is 7; ori is 3, m is 10, and n is 5; or

[0100] i is 13, m is 10, and n is 5; or

[0101] i is 15, m is 10, and n is 5; or

[0102] i is 11, m is 10, and n is 5; or

[0103] when L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0104] i is an integer selected from 0 to 20;

[0105] m is an integer selected from 0 to 15; and

[0106] n is an integer selected from 0 to 15.

[0107] In some embodiments, the compound of Formula (I) is an enantiomer as represented by Formula (1-1):

[0108]

[0109] In some embodiments, the compound of Formula (I) is selected from

[0110] O

[0111]

[0112] wherein

[0113] i is 15, m is 10, and n is 5; or

[0114] i is 11, m is 10, and n is 5.

[0115] In some embodiments, the compound of Formula (I) is selected from

[0116] - XH / x / , " N

[0117] 'i N O

[0118]

[0119] i is an integer selected from 3 to 20;

[0120] m is an integer selected from 0 to 15; and

[0121] n is an integer selected from 0 to 15.

[0122] In some embodiments, the disease or condition associated with lipid accumulation isselected from a metabolic disease and / or an inflammatory disease.

[0123] In some embodiments, the disease or condition associated with lipid accumulation is selected from dyslipidemia, hypertriglyceridemia, mixed dyslipidemia, obesity, insulin resistance, impaired glucose tolerance, type 2 diabetes, metabolic syndrome, visceral adiposity, fatty liver disease including metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), alcohol-associated fatty liver disease, steatotic liver with fibrosis, liver cirrhosis secondary to steatosis, pancreatic steatosis, skeletal muscle steatosis, cardiac steatosis, atherosclerotic cardiovascular disease, hyperlipidemia-associated hypertension, imbalanced gut microbiome, increased intestinal permeability, inflammatory bowel diseases including ulcerative colitis and Crohn's disease, microbiome-associated inflammatory conditions, chronic low-grade inflammation associated with ectopic lipid deposition, and inflammatory diseases associated with dyslipidemia and metabolic dysfunction.

[0124] In some embodiments, the disease or condition associated with lipid accumulation is a multifactorial disease.

[0125] In some embodiments, the method is characterised by a tolerance to insulin and / or a tolerance to glucose absorption.

[0126] Brief description of the drawings

[0127] Embodiments of the present invention will now be described, by way of non-limiting example, with reference to the drawings in which:

[0128] Figure 1A-C illustrates enzymatic and chemical methods to synthesis FAHFAs. A: Examples of commercially interesting FAHFA. B: Current enzymatic synthesis of unique FAHFAs. C: Current chemical synthesis of unique FAHFAs.

[0129] Figure 2A illustrates the synthesis of FAHFA. (a) alkylmagnesium bromide (3.0 equiv.), THF, 0 °C to rt, 45-83% yield; (b) SOCI2 (1.3 equiv.), MeOH, reflux, 90-95% yield; (c) acyl chloride (1.5 equiv.), pyridine (5 equiv.), CH2CI2, 0 °C to 23 °C, 74-85% yield; (d) LiOH·H2O (6.5 equiv.), THF / H2O (1:1, v / v), 0 °C to 23 °C, 88–93% yield.

[0130] Figure 2B illustrates 49 members of the FAHFA family and 15 ester bond bioisosteres there were synthesised.

[0131] Figure 3A shows lipid absorption screens using 51 FAHFAs. Volcano plots showing thelipid absorption screens of 51 structurally different FAHFAs in the absence (left) and presence (right) of insulin stimulation. The screen results are expressed in Iog2 fold change of lipid absorption rate relative to DMSO-treated cells. FAHFAs were shortlisted based on p-value <0.1 and Iog2 fold change < -0.8. Shortlisted FAHFAs and the corresponding concentration are annotated (e.g. 12-SAHSA_100 represents 100 pg / mL of 12-SAHSA).

[0132] Figure 3B shows a Venn diagram of 12 FAHFAs from lipid absorption screens with and without insulin stimulation.

[0133] Figure 4A shows dose-response curves of the 12 shortlisted FAHFAs in relation to lipid absorption rate of HepG2 upon exposure to 1.56-200 pg / mL of the respective FAHFAs. Dotted line indicates the baseline of DMSO-treated control. Data are presented as mean ± SD from three replicates.

[0134] Figure 4B shows dose-response curves of the 12 shortlisted FAHFAs in relation to glucose absorption of HepG2 upon exposure to 1.56-200 pg / mL of the respective FAHFAs. Dotted line indicates the baseline of DMSO-treated control. Data are presented as mean ± SD from three replicates.

[0135] Figure 4C shows dose-response curves of the 12 shortlisted FAHFAs in relation to viability of HepG2 upon exposure to 1.56-200 pg / mL of the respective FAHFAs. Data are presented as mean ± SD from three replicates.

[0136] Figure 5 shows lipid absorption dose-response curves of HepG2 upon exposure to 12-SAHSA and 12-TAHSA derivatives. HepG2 were treated with 1.56-200 pg / mL of the respective FAHFAs. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the lipid absorption rate of DMSO-treated control. The absence and presence of insulin stimulation are indicated by grey (#) and black (*) lines, respectively. Data are presented as mean ± SD from three replicates.

[0137] Figure 6 shows lipid absorption dose-response curves of AML12 upon exposure to 12-SAHSA and 12-TAHSA derivatives. AML12 were treated with 1.56-200 pg / mL of the respective FAHFAs. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the lipid absorption rate of DMSO-treated control. The absence and presence of insulin stimulation are indicated by grey (#) and black (*) lines, respectively. Data are presented as mean ± SD from three replicates.

[0138] Figure 7 shows glucose absorption dose-response curves of HepG2 upon exposure to 12-SAHSA and 12-TAHSA derivatives. HepG2 were treated with 1.56-200 pg / mL of the respective FAHFAs. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the glucose absorption of DMSO-treated control. The absence and presence of insulin stimulation are indicated by grey (#) and black (*) lines, respectively. Data are presented as mean ± SD from three replicates.Figure 8 shows glucose absorption dose-response curves of AML12 upon exposure to 12-SAHSA and 12-TAHSA derivatives. AML12 were treated with 1.56-200 pg / mL of the respective FAHFAs. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the glucose absorption of DMSO-treated control. The absence and presence of insulin stimulation are indicated by grey (#) and black (*) lines, respectively. Data are presented as mean ± SD from three replicates.

[0139] Figure 9 shows viability of HepG2 upon exposure to 12-SAHSA and 12-TAHSA derivatives. HepG2 were treated with 1.56-200 pg / mL of the respective FAHFAs for 48 h. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the baseline viability of DMSO-treated control. Data are presented as mean ± SD from three replicates.

[0140] Figure 10 shows viability of AML12 upon exposure to 12-SAHSA and 12-TAHSA derivatives. AML12 were treated with 1.56-200 pg / mL of the respective FAHFAs for 48 h. FAHFAs highlighted in a box are the parental structures. Dotted line indicates the baseline viability of DMSO-treated control. Data are presented as mean ± SD from three replicates.

[0141] Figure 11A-C shows in vivo efficacy of FAHFAs in a diet-induced obese mouse model. Weight gain (A), the area under curve of intraperitoneal glucose tolerance test (IGTT AUC; (B), the normalized weight (C) of mice on control diet, LIDPAD (high-calorie diet) or those treated with semaglutide (100 pg / kg; twice per week), 12-SAHSA, 12-TAHSA, 12-SAASA, 12-TAASA or 12-HDTZSA (lOOmg / kg / day) while on LIDPAD feeding for 28 days. * p<0.05, ** p<0.01, *** p<0.001. Data are presented as mean ± SD. Sample size is 4-6 mice per group.

[0142] Figure 11D shows representative histological images of liver and epididymal white adipose tissues (eWAT) of mice on control diet, LIDPAD (high-calorie diet) or those treated with semaglutide (100 pg / kg; twice per week), 12-SAHSA, 12-TAHSA, 12-SAASA, 12-TAASA or 12-HDTZSA (lOOmg / kg / day) while on LIDPAD feeding for 28 days. * p<0.05, ** p<0.01, *** p<0.001. The scale bar is 200 pm.

[0143] Figure 12A-D shows the impact of selected FAHFAs on other organs. Normalized weight of heart (A), spleen (B), left and right kidneys (C 8i D) of mice on control diet, LIDPAD (high-calorie diet) or those treated with semaglutide (100 pg / kg; twice per week), 12-SAHSA, 12-TAHSA, 12-SAASA, 12-TAASA or 12-HDTZSA (lOOmg / kg / day) while on LIDPAD feeding for 28 days. N. S. not significant. Data are presented as mean ± SD. Sample size is 4-6 mice per group.

[0144] Figure 13A-B shows 12-TAASA and 12-HDTZSA are potential microbiome modulators. (A) Mass of parental FAHFAs (12-SAHSA & 12-TAHSA) and their derivatives (12-SAASA, 12-TAASA & 12-HDTZSA) present in mouse fecal samples 24 h and 48 h after a singledose FAHFA oral administration. (B) Beta diversity analysis of 16S gut microbial profiles.

[0145] Figure 13C shows relative abundance of key beneficial and pathogenic genera from different treatment groups.

[0146] Figure 13D-E shows (D) Functional changes related to short-chain fatty acids (SCFA) synthesis predicted based on gut microbial profiles. The pathways were stratified into fermentation- and lipid-related depending on the primary substrates used in the SCFA biosynthetic pathways. Log2 (Fold change) were coded in patterns and expressed in relative to the control diet group. (E) Relative abundance of predominant SCFA producers in the gut microbiome (Blautia, Lachnospiraceae NK4A136 group and Roseburia) between different treatment groups. Data are presented as mean ± SD from at least 3 biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001.

[0147] Figure 14A-B shows 12-TAASA disrupts intestinal lipid uptake via perturbation of CD44-associated gene network. (A) Principal component analysis (PCA) of the intestinal epithelial transcriptomes of mice supplemented with different synthetic FAHFAs and untreated LIDPAD mice. (B) Functional enrichment analysis comparing intestinal epithelial transcriptomes between FAHFA-treated mice and untreated LIDPAD mice. Expression patterns of the gene sets represent the activation status of the enriched relative to untreated LIDPAD mice.

[0148] Figure 14C shows expression pattern of genes (bottom panel; patterned = downregulation; black = upregulation) involved in various lipid metabolic processes (top panel; grey box indicates the involvement of a gene in a pathway).

[0149] Figure 14D-G shows (D) Top-ranked hub genes identified within the gene-gene interaction networks of downregulated DEGs from intestinal epithelial transcriptomes in 12-TAASA-treated mice compared to untreated LIDPAD mice. Hub genes were prioritized based on their betweenness centrality and degree centrality scores. The size of each node reflect the statistical significance. (E) Chemical structure of biotinylated 12-TAASA used in a pull-down assay. (F) Top 5 enriched biological functions of 134 putative 12-TAASA binding membrane proteins in the intestinal epithelial lysates. (G) Venn diagram showing three shared genes / gene products between the 12-TAASA-binding membrane proteins and genes in the intestinal epithelium suppressed by 12-TAASA.

[0150] Figure 15 shows the acid stability of FAHFA derivatives, in percentage improvement in acid stability of synthetic FAFAH derivatives compared to the parental 12-SAHSA. Figure 16A-B shows structure-activity relationship of FAHFAs in cellular lipid uptake study. Pearson correlation analysis of AUC (Area Under the Curve) of lipid uptake in the presence of insulin and at basal condition. Using an arbitrary cutoff of AUC < -25, FAHFAs exhibiting lipid uptake inhibition in an insulin insensitive (round nodes), insulinsensitive (triangle nodes) manner or have no inhibition effect on lipid uptake (square nodes) are annotated. (A) The nodes are shape-coded based on the branching carbon position; (B) to highlight regioisomers that exhibit suppressive effect on cellular lipid uptake.

[0151] Figure 16C-D shows (C) AUC of cellular lipid and glucose uptake and viability assays following treatment of 7 shortlisted FAHFAs across a concentration range of 1.56-200 pg / mL. The x-axis (bottom) indicates the AUC of glucose (solid circles) and lipid (hollow circles) uptake where negative values represent stronger inhibition while the secondary x-axis (top) indicates the AUC of viability where positive values represent higher viability; (D) AUC of cellular lipid uptake following treatment of FAHFAs with different branching carbon positions.

[0152] Figure 17A shows screening of FAHFA derivatives with ester bond bioisosteres. Scatter plots showing AUC of lipid uptake assays following treatments of parental FAHFAs (12-SAHSA and 12-TAHSA) and their derivatives across a concentration range of 1.56-200 pg / mL in two hepatocyte cell lines, HepG2 and AML12. The assays were performed at basal condition and with insulin stimulation. 12-TAASA, 12-SAASA and 12-HDTZSA were shortlisted for in vivo testing.

[0153] Figure 17B shows AUC of glucose (solid circles) and lipid (hollow circles) uptake (bottom x-axis; negative values indicate stronger inhibition) and viability (top x-axis; positive values indicate higher viability) in HepG2 or AML12 cells treated with FAHFA derivatives (1.56-200 pg / mL). Acid stability (percentage remaining at pH 1.5 after 12h; black scale and lines).

[0154] Figure 18A-B shows CD44-associated gene network is a molecular target of 12-TAASA. (A) Molecular docking of 12-TAASA and human CD44 hyaluronic acid binding domain (CD44-HABD) using two algorithms, Attracting Cavities 2.0 (left) and Autodock Vina (right). The interacting amino acid residues and the docking / affinity scores are annotated. (B) Microscopic images of lipid absorption (left) in CD44 knockdown-NCM460 with / without 12-TAASA (100 pg / mL) treatment and quantification of the lipid absorption based on Bodipy intensity (right top). Knockdown efficiency of CD44 (right bottom) is demonstrated using immunoblotting. ***p<0.001.

[0155] Detailed description

[0156] " Alkyl" refers to monovalent alkyl groups which may be straight chained or branched and preferably have from 1 to 10 carbon atoms or more preferably 1 to 6 carbon atoms. Examples of such alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, n-hexyl, and the like." Aryl" refers to an unsaturated aromatic carbocyclic group having a single ring (eg. phenyl) or multiple condensed rings (e.g. naphthyl or anthryl), preferably having from 6 to 14 carbon atoms. Examples of aryl groups include phenyl, naphthyl and the like.

[0157] " Cycloalkyl" refers to cyclic alkyl groups having a single cyclic ring or multiple condensed rings, preferably incorporating 3 to 11 carbon atoms. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, indanyl, 1,2,3,4-tetrahydronapthalenyl and the like.

[0158] " Oxylene" refers to a divalent group -O-.

[0159] " Oxyacylene" refers to a divalent group -OC(O)- or -C(O)O-.

[0160] " Oxyacyloxyene" refers to a divalent group -OC(O)O-.

[0161] " Aminoacylene" refers to a divalent group -C(O)NR- where R is selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl.

[0162] " Oxysulfonylene" refers to a divalent group -OS(O)2- or -S(O)2O-.

[0163] " Heteroaryl" refers to a monovalent aromatic heterocyclic group which fulfils the Hiickel criteria for aromaticity (i.e. contains 4n + 2 n electrons) and preferably has from 2 to 10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen, selenium, and sulfur within the ring (and includes oxides of sulfur, selenium and nitrogen). Such heteroaryl groups can have a single ring (eg. pyridyl, pyrrolyl or N-oxides thereof or furyl) or multiple condensed rings (eg. indolizinyl, benzoimidazolyl, coumarinyl, quinolinyl, isoquinolinyl or benzothienyl).

[0164] Examples of heteroaryl groups include, but are not limited to, oxazole, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, isothiazole, phenoxazine,phenothiazine, thiazole, thiadiazoles, oxadiazole, oxatriazole, tetrazole, thiophene, benzo[b]thiophene, triazole, imidazopyridine and the like.

[0165] " Heteroarylene" refers to a divalent aryl group wherein the heteroaryl group is as described above.

[0166] " Heterocyclyl" refers to a monovalent saturated or unsaturated group having a single ring or multiple condensed rings, preferably from 1 to 8 carbon atoms and from 1 to 4 hetero atoms selected from nitrogen, sulfur, oxygen, selenium or phosphorous within the ring. The most preferred heteroatom is nitrogen. It will be understood that where, for instance, R2or R' is an optionally substituted heterocyclyl which has one or more ring heteroatoms, the heterocyclyl group can be connected to the core molecule of the compounds of the present invention, through a C-C or C-heteroatom bond, in particular a C-N bond.

[0167] Examples of heterocyclyl groups include, but are not limited to, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1, 2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazolidine, morpholino, pyrrolidine, tetra hydrofuranyl, and the like.

[0168] " Heterocyclylene" refers to a divalent heterocyclyl group wherein the heterocyclyl group is as described above.

[0169] Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers. " Optically-enriched," as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound of the present invention is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments the compound is made upof at least about 95%, 98%, or 99% by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ, of Notre Dame Press, Notre Dame, IN 1972).

[0170] Fatty acid esters of hydroxy fatty acids (FAHFAs) are a class of naturally occurring bioactive lipids characterized by an ester linkage between a fatty acid and a hydroxyfatty acid. Discovered in 2014, FAHFAs were recognized for their potent antiinflammatory and metabolic regulatory properties, quickly garnering significant attention due to their therapeutic potential for metabolic disorders, inflammatory diseases, and certain cancers. Notably, specific FAHFA species such as 5-PAHSA, 9-PAHSA, 13-DHAHLA, and 13-LAHLA have demonstrated robust biological activities, including improvement of insulin sensitivity, modulation of glucose metabolism, and suppression of pro-inflammatory cytokine secretion.

[0171] The physiological significance of FAHFAs is underscored by their endogenous presence in various organisms and dietary sources, including plants, animals, and common foods like breast milk, tea, olive oil, and nuts. Importantly, individuals with insulin resistance and obesity-related metabolic diseases exhibit lower circulating FAHFA levels, highlighting their crucial role in maintaining metabolic homeostasis and immune regulation. Supplementation of specific FAHFAs in animal models has effectively reversed metabolic dysfunction, indicating their promise as potential oral therapeutics for obesity and diabetes.

[0172] Despite their considerable therapeutic potential, progress in FAHFA research has been unacceptably slow. Food sources contain hundreds of structurally diverse FAHFAs; however, their extremely low abundances significantly hinder systematic screening and comprehensive biological characterization. Additionally, the synthesis of these molecules is inherently challenging and expensive, involving multi-step chemical routes with poor yields and limited scalability. Recent advances in lipid chemistry have offered potential solutions, exploring both enzymatic and chemical methods for FAHFA production. For instance, a bi-enzymatic cascade has been developed to convertrenewable unsaturated fatty acids into optically pure FAHFAs. Several chemical synthesis approaches have also been reported, typically involving 3 to 7 steps from substrates like alkenes, epoxides, and aldehydes. 5-FAHFA was first synthesised in 2014, though the process suffered from low yield and atom economy. Another process utilized epoxide-opening routes to generate various FAHFA homologues, although the final step required toxic chromium reagents. Another method employed epoxy chloropropane and alkenyl precursors to obtain (R)-9-PAHSA enantiomers. While these efforts expanded accessible FAHFA analogues, issues of low efficiency, hazardous reagents, and limited structural diversity persisted. Furthermore, improving the ester bond stability under gastrointestinal conditions remained a key unmet need to enable oral delivery. Moreover, the ester bonds in native FAHFAs are highly susceptible to enzymatic hydrolysis and acidic degradation in the gastrointestinal tract, severely compromising their oral bioavailability and stability. These limitations pose critical barriers to their practical development and widespread therapeutic application.

[0173] Addressing these challenges require innovative synthetic strategies capable of efficient, scalable production and chemical stabilization of FAHFA derivatives. The present disclosure relates to a modular, scalable chemical synthesis platform specifically engineered to overcome the existing synthetic complexities and stability constraints of native FAHFAs. This versatile platform facilitates the production of a diverse array of FAHFA analogues incorporating chemically stable linkages such as amide (-CONH-), sulfonate (-OSO2-), ether (-O-), and 1,2,3-triazole, effectively enhancing their resistance to enzymatic and acidic degradation. Through this robust synthetic approach, the inventors identified FAHFA analogues exhibiting potent inhibition of intestinal lipid absorption, dual mechanism-of-action involving host intestinal pathways and microbiome remodeling, and significant therapeutic efficacy in diet induced obesity mouse models. Collectively, this modular synthesis platform advances the translational potential of FAHFAs, offering a new class of chemically stable, orally bioavailable therapeutics for obesity and associated metabolic diseases.

[0174] Metabolic dysfunction-associated steatotic liver disease (MASLD) is now the world's most common chronic liver condition and a leading cause of cirrhosis, hepatocellular carcinoma, and transplantation. Despite recent approvals for MASH with fibrosis, most interventions act late, after hepatic injury is entrenched. A focus of this invention is upstream, on the gut-liver axis, where excessive gut to-liver lipid flux is a defining driver of disease. Enterocytes absorb and package dietary fats into chylomicrons that continuously load the liver, straining storage and stress programs to seed steatosis andinflammation. This flux is not fixed. Microbiome-dependent signals, bile acid composition, short-chain fatty acids (SCFA), and epithelial barrier integrity, set the gain on intestinal lipid delivery. Positioning the intestine as a proximal, druggable checkpoint reframes MASLD therapy. By modulating absorption and the microbial milieu, hepatic lipid input can be decreased before irreversible injury accrues. This gut-first strategy is broadly applicable, mechanistically anchored, and primed to complement emerging antifibrotics.

[0175] To address MASLD at its proximal driver, excess gut-to-liver lipid flux, FAHFA-based agents that are chemically stabilized for oral delivery yet retain (or enhance) intestinecentric and microbiome-linked biology were sought. Here, a modular, scalable platform that installs bioisosteric linkages to generate orally stable FAHFA derivatives (Figure 16A-B) is developed. Accordingly, an intestine-anchored mechanism that lowers gut-to-liver lipid flux by suppressing intestinal lipid absorption and biasing the microbiome toward SCFA producers has been uncovered, positioning stabilized FAHFAs as a tractable entry point for MASLD.

[0176] The present disclosure concerns fatty acid esters of hydroxy fatty acids (FAHFAs), their analogs and / or derivatives, and their synthesis thereof. The FAHFAs may be used in inhibiting the absorption of lipids and its usage to prevent obesity- related conditions. This has applications for the development of cheaper and potentially safer anti-obesity interventions.

[0177] In particular, the inventors had synthesised and screened naturally occurring and bioisosteres of FAHFAs and identified specific FAHFA derivatives that inhibit lipid absorption and limit weight gain in diet-induced obese models. These FAHFAs and their derivatives have an inhibitory effect on fat absorption for weight loss. Their efficacy, safety profile and mechanism of action were validated through systematically designed cell culture experiments and a robust diet-induced obese mouse model.

[0178] It was found that the ester bond connecting the fatty acid and the hydroxy fatty acid functional groups was susceptible to degradation by acid hydrolysis, negatively impacting its chemical stability during oral administration. By bioisosteric replacement of the labile ester bond in natural FAHFAs to analogs, the compounds demonstrate better stability and prolonged bioactivities. This approach directly addresses the rapid degradation of FAHFAs in the acidic gastric environment, enhancing their oral bioavailability. In this regard, unlike natural FAHFAs, the analogs (e.g., carbonateesters, sulfonates, amides, triazoles) are more resistant to acid hydrolysis, optimizing their delivery through oral intake. These compounds may be synthesized rapidly through a modular platform that allows for efficient production of multiple novel classes of FAHFA analogs (carbonates, sulfonates, amides, triazoles).

[0179] The library of FAHFA analogs provides a unique toolset to systematically dissect how the chemical nature of the linkage influences hydrolytic stability and biological activity, offering new insights beyond the native ester-based FAHFAs. Furthermore, by offering new FAHFA derivatives that are chemically stable, the pharmacokinetic limitations of natural FAHFAs may be overcome to unlock robust and durable efficacy through oral delivery which is more accessible and acceptable compared to other administration route. Such feature of the modified FAHFAs is valuable, especially in chronic health conditions where long-term treatment is essential.

[0180] Accordingly, the present disclosure relates to a method of synthesising a compound of Formula (I), or a salt, solvate, prodrug, stereoisomer or derivative thereof, comprising:

[0181]

[0182] wherein

[0183] L is a linker selected from oxylene, oxyacylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0184] m is an integer selected from 0 to 20;

[0185] n is an integer selected from 0 to 20; and

[0186] i is an integer selected from 0 to 20.

[0187] The compounds of the present disclosure may be synthesized by a 1,2 addition of a nucleophile to the electrophilic carbon of the aldehyde. For example, a Grignard reagent may be 1,2 added to an aldehyde, producing various hydroxy-fatty acids. In this reaction, the nucleophilic pair of electrons in the C-Mg bond attach the carbonyl carbon, breaking the C-0 bond. The negatively charged O is then protonated to give the compound of Formula (IV), which typically comprises a chiral center. The reaction may thus form a racemic mixture.

[0188]

[0189] (II) (III)The hydroxy-fatty acids may then be esterified at its carboxylic acid moiety to form hydroxy-fatty acid methyl esters.

[0190] OH OH O

[0191]

[0192] (IV) (V)

[0193] Esterification of the secondary alcohol with the appropriate reagent leads to the formation of a protected compound of Formula (VI).

[0194] OH O

[0195] o'Rl

[0196]

[0197] (V) (VI)

[0198] Finally, selective hydrolysis of the more accessible methyl ester yields compounds of Formula (I). Using this approach, 49 FAHFAs (L is oxyacylene) were synthesised. In particular, this route allows for the synthesis of compounds of Formula (I) in which L is an ester moiety, sulfonate moiety, carbonate moiety, or ether moiety. These compounds may be represented as:

[0199]

[0200] The compound of Formula (I) with L being an amide moiety may be synthesised starting from a compound of Formula (V). For example, the compound of Formula (V) may proceed through a Dess-Martin oxidation followed by a reductive amination sequence to form compound of Formula (VIII). Dess-Martin Periodinane (DMP), a hypervalentiodine compound, can provide selective and very mild oxidation of alcohols to aldehydes or ketones. The oxidation may be performed in dichloromethane or chloroform at room temperature, for about 0.5 h to about 2 h. Reductive amination is a form of amination that converts a carbonyl group (a ketone or an aldehyde) to an amine via an intermediate imine. Finally, compound of Formula (VIII) is subjected to amidation to form compound of Formula (VIb).

[0201] NaBH3CN

[0202] OH O Dess-Martin MU O H,K1, R2^

[0203] O O

[0204] (V) (XIV)

[0205]

[0206] The compound of Formula (I) with L being a heterocyclyl or heteroaryl may be synthesised starting from a compound of Formula (V). For example, to form a compound with L being a triazole moiety, compound of Formula (V) undergoes bromination (Appel reaction) followed by azidation to afford a compound of Formula (IX). Finally, the compound of Formula (IX) is subjected to a click reaction to form compound of Formula (Vic).

[0207]

[0208] The present disclosure comprises within its scope, both the R and S stereoisomer of compound of Formula (I), as represented by the Markush structure of Formula (I). For example, the chiral center may be at the carbon linked to the L moiety. Formula (I) may alternatively be represented as Formula (I'):

[0209]

[0210] OH are represented as:

[0211]

[0212] The skilled person would know of various synthetic processes to synthesise a stereoisomer of Formula (I). For example, by selecting a chiral Grignard reagent, the reaction may be stereoselective through the transfer for chirality from the Grignard reagent to the carbon. The subsequent reactions may be selectively performed to retain this stereochemistry.

[0213] For example, the compounds of the present disclosure may be an enantiomer synthesized by selective addition of a chiral Grignard reagent to an aldehyde, producing stereoisomer (IV-1).

[0214] OH

[0215] ^MgX - - chiral ligand

[0216]

[0217] (IV-1)

[0218] The stereoisomer (IV-l)may then be esterified at its carboxylic acid moiety to form hydroxy-fatty acid methyl esters.

[0219] OH O

[0220]

[0221] (V-1)

[0222] Esterification of the secondary alcohol with the appropriate reagent leads to the formation of a protected compound of Formula (VI-1).

[0223] OH O

[0224] o'Rl

[0225]

[0226] (V-1) (VI-1)

[0227] For example, selective hydrolysis of the more accessible methyl ester yields a compound of Formula (1-1). This route allows for the synthesis of compounds of Formula (I) in which L is an ester moiety, sulfonate moiety, carbonate moiety, and ether moiety. Thesecompounds may be represented as:

[0228] (1-1)

[0229] (la-D

[0230] (Id-1)

[0231] (le-D

[0232]

[0233] (If-1).

[0234] The enantiomer of Formula (I) with L being a heterocyclyl or heteroaryl may be synthesised starting from a compound of Formula (V-l). For example, to form a compound with L being a triazole moiety, compound of Formula (V-l) undergoes bromination (Appel reaction) followed by azidation to afford compound of Formula (IX- 1). Finally, compound of Formula (IX-1) is subjected to a click reaction to form compound of Formula (Vic).

[0235] (V-1) (IX-1)

[0236]

[0237] (lx'1) (X) (Vlc-1)

[0238] Alternatively, as shown in Procedure D, chirality may be imparted through the use of a biocatalyst. The subsequent reactions may be performed to selectively retain this stereochemistry.As a corollary, the stereoisomer of Formula (1-2) may also be synthesized by varying the synthetic protocol disclosed herein, by using suitable reactants and / or reagents.

[0239] Thus, the present disclosure relates to a method of synthesising a compound of Formula (I), or a salt, solvate, prodrug, stereoisomer or derivative thereof, comprising:

[0240]

[0241] wherein

[0242] L is a linker selected from oxylene, oxyacylene, aminoacylene, oxysulfonylene. oxyacyloxyene, heterocyclylene, and heteroarylene;

[0243] m is an integer selected from 0 to 20;

[0244] n is an integer selected from 0 to 20;

[0245] i is an integer selected from 0 to 20;

[0246] ai) conjugating an alkyl moiety via linker to the hydroxy-alkyl ester of Formula (V) in order to form a compound of Formula (VI);

[0247] OH O O

[0248] ““ MWSO-R’

[0249]

[0250] (V) (VI)

[0251] wherein Ri is optionally substituted alkyl; and

[0252] b) hydrolysing the ester in order to form the compound of Formula (I).

[0253] In some embodiments, the compound of Formula (I) is a compound of Formula (1-1):

[0254]

[0255] In some embodiments, step a) comprises an esterification step of the secondary alcohol. In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (VII) to form a compound of Formula (Via).

[0256] OH O

[0257] (V)

[0258]

[0259] wherein X is halo.In some embodiments, X is F, Cl or Br. In some embodiments, X is Cl.

[0260] In some embodiments, step a) comprises converting a compound of Formula (V) into a compound of Formula (VIII) and with a compound of Formula (VII) to form a compound of Formula (Vlb).

[0261]

[0262] (VII) (Vlb) wherein R2 is independently selected from H and optionally substituted alkyl; and X is halo.

[0263] In some embodiments, R2 is independently selected from H, methyl and ethyl. In some embodiments, R2 is H.

[0264] In some embodiments, X is F, Cl or Br. In some embodiments, X is Cl.

[0265] In some embodiments, step a) comprises a reductive amination step to convert compound of Formula (XIV) to the compound of Formula (VIII).

[0266] NaBH3CN

[0267] NH2-R2

[0268]

[0269] (XIV) (VIII)

[0270] The reducing agent may be sodium cyanoborohydride (NaBF CN), or NaBH4, or NaBH(OAc)3. The amine compound may be ammonium acetate (NF Ac).

[0271] In some embodiments, step a) comprises an oxidation step to convert the hydroxyl of compound of Formula (V) into an acyl moiety (carbonyl). The oxidation step may occur via a Dess-Martin oxidation. The oxidation may be performed in dichloromethane or chloroform at room temperature, for about 0.5 h to about 2 h.

[0272] Dess-Martin

[0273]

[0274] (V) (XIV)In some embodiments, step a) comprises a click coupling step coupling a compound of Formula (IX) with a compound of Formula (X) to form a compound of Formula (Vic). The azide-alkyne cycloaddition (CuAAC) may be catalysted by a copper catalyst, such copper(II) sulfate pentahydrate, copper(II) sulfate, copper(I) iodide, copper(I) bromide, and copper(II) acetate (Cu(0Ac)2).

[0275] / / " CU(OAC)2, N

[0276] N O

[0277] 'HM '

[0278]

[0279] (IX) (Vic)

[0280] In some embodiments, step a) further comprises a bromination step followed by azidation step to convert the compound of Formula (V) to the compound of Formula (IX).

[0281] OH O PPH3, CBr4Br 0

[0282]

[0283] In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XI) to form a compound of Formula (Vid).

[0284] OH O

[0285]

[0286] (XI) (Vid) wherein X is halo.

[0287] In some embodiments, X is F, Cl or Br. In some embodiments, X is Cl.

[0288] In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XII) to form a compound of Formula (Vie).

[0289] < X,0 OH O o

[0290]

[0291] (XII) (Vie) wherein X is halo.

[0292] In some embodiments, X is F, Cl or Br. In some embodiments, X is Cl.In some embodiments, step a) comprises reacting a compound of Formula (V) with a compound of Formula (XIII) to form a compound of Formula (VIf).

[0293] O

[0294]

[0295] (V) (XIII)(V|f) wherein X is halo.

[0296] In some embodiments, X is F, Cl or Br. In some embodiments, X is Cl.

[0297] In some embodiments, Ri is optionally substituted C1-C5 alkyl. In some embodiments, Ri is methyl, ethyl, or propyl. In some embodiments, Ri is methyl.

[0298] In some embodiments, step a) further comprises a step of esterifying a compound of Formula (IV) to form the compound of Formula (V). The compound of Formula (IV) may be esterified at its carboxylic acid moiety using an alkyl alcohol and in the presence of an acid or base catalyst. The alkyl alcohol may for example be MeOH.

[0299]

[0300] (IV) (V)

[0301] In some embodiments, step a) further comprises a step of reacting a compound of Formula (II) with a compound of Formula (III) to form the compound of Formula (IV). The compound of Formula (III) is a Grignard reagent (organomagnesium halide) which acts as a potent nucleophile / strong base to form a carbon-carbon bond with the compound of Formula (II).

[0302] OH

[0303] o^uCOOH* "M9X’

[0304]

[0305] (II) (III) (IV) wherein X is halo, preferably Br.

[0306] The reaction may be performed under inert conditions such as under nitrogen. The reaction may be performed at 0 °C. The reaction may be performed for about 6 h, or about 8 h at room temperature.In some embodiments, step b) is performed in the presence of an alkali. In some embodiments, the alkali is a strong alkali such as LiOH, NaOH, or KOH.

[0307] In some embodiments, step b) is performed for at least 12 h, or 24 h.

[0308] In some embodiments, the compound of Formula (I) is a compound of Formula (la-f):

[0309]

[0310] wherein

[0311] i is an integer selected from 3 to 20;

[0312] m is an integer selected from 0 to 15; and

[0313] n is an integer selected from 0 to 15.

[0314] In some embodiments, the compound of Formula (I) is a compound of Formula (la-c):

[0315] O

[0316]

[0317] (la)O

[0318] (Ib)

[0319]

[0320] (Ic)

[0321] wherein

[0322] i is an integer selected from 3 to 20;

[0323] m is an integer selected from 0 to 15; and

[0324] n is an integer selected from 0 to 15.

[0325] In some embodiments, when L is oxyacylene,

[0326] i is an integer selected from 0 to 15;

[0327] m is an integer selected from 5 to 10; and

[0328] n is an integer selected from 5 to 10.

[0329] In some embodiments, i is an integer selected from 0 to 15, 5 to 15, 7 to 15, 8 to 15, 9 to 15, 10 to 15, or 11 to 15.

[0330] In some embodiments, m is an integer selected from 5 to 10, 7 to 10, 8 to 10, or 9 to 10. In some embodiments, m is 10.

[0331] In some embodiments, n is an integer selected from 5 to 9, 5 to 8, 5 to 7, or 5 to 6. In some embodiments, n is 5.

[0332] In some embodiments, when L is oxyacylene, i is 15, m is 8, and n is 7. This corresponds to 10-SAHSA.

[0333] In some embodiments, when L is oxyacylene, i is 3, m is 10, and n is 5. This corresponds to 12-HAHSA.

[0334] In some embodiments, when L is oxyacylene, i is 13, m is 10, and n is 5. This corresponds to 12-PAHSA.

[0335] In some embodiments, when L is oxyacylene, i is 15, m is 10, and n is 5. This corresponds to 12-SAHSA.In some embodiments, when L is oxyacylene, i is 11, m is 10, and n is 5. This corresponds to 12-TAHSA.

[0336] In some embodiments, when L is a linker selected from aminoacylene, heteroarylene, oxylene, oxysulfonylene, oxyacyloxyene, and heterocyclylene;

[0337] i is an integer selected from 0 to 20;

[0338] m is an integer selected from 0 to 15; and

[0339] n is an integer selected from 0 to 15.

[0340] In some embodiments, i is an integer selected from 3 to 20, 5 to 20, 7 to 20, 10 to 20, 10 to 18, 10 to 16, or 10 to 15. In some embodiments, m is an integer selected from 0 to 15, 1 to 15, 3 to 15, 5 to 15, 7 to 15, 9 to 15, 10 to 15, 10 to 14, 10 to 13 or 10 to 12. In some embodiments, m is 10. In some embodiments, n is an integer selected from 0 to 15, 1 to 15, 1 to 13, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 2 to 5, or 3 to 5. In some embodiments, n is 5.

[0341] In some embodiments,

[0342] i is an integer selected from 3 to 20;

[0343] m is an integer selected from 0 to 15; and

[0344] n is an integer selected from 0 to 15.

[0345] In some embodiments,

[0346] i is an integer selected from 10 to 16;

[0347] m is an integer selected from 10 to 14; and

[0348] n is an integer selected from 1 to 6.

[0349] In some embodiments, when L is aminoacylene, I is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, i is 15, m is 10 and n is 5. This corresponds to 12-SAASA. In some embodiments, i is 11, m is 10 and n is 5. This corresponds to 12-TAASA.

[0350] In some embodiments, when L is heteroarylene, i is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, I is 14, m is 10 and n is 5. This corresponds to 12-HDTZSA.

[0351] In some embodiments, when L is oxylene, i is an integer selected from 10 to 16, m is10 and n is 5.

[0352] In some embodiments, when L is oxysulfonylene, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0353] In some embodiments, when L is oxyacyloxyene, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0354] In some embodiments, the method further comprises a step of deuterating the compound of Formula (I) or intermediate thereof. For example, the compound of Formula (IV) may be oxidised to a compound of Formula (XIV) and reduced in the presence of a deuterated reducing agent, for example, NaBD4.

[0355] Accordingly, in some embodiments, the compound of Formula (I) is deuterated. In some embodiments, the compound of Formula (I) is deuterated at at least one H position. In some embodiments, the compound of Formula (I) is deuterated at at least two, three, four or five H positions. In some embodiments, the compound of Formula (I) is deuterated at all H positions. In some embodiments, the compound of Formula (I) is at an alpha position relative to L.

[0356] In some embodiments, step a) comprises conjugating a halo substituted alkyl moiety via linker to the hydroxy-alkyl ester of Formula (V) in order to form a halogenated compound of Formula (VI).

[0357] Accordingly, in some embodiments, the compound of Formula (I) is halogenated. In some embodiments, the compound is halogenated on its fatty acid ester chain. In some embodiments, the compound is halogenated on its hydroxy fatty acid backbone. In some embodiments, the compound is halogenated at one, two, three, four or five positions. In some embodiments, the compound is halogenated at all C positions on its fatty acid ester chain.

[0358] In some embodiments, step a) is a step of stereoselectively reacting the compound of Formula (V) to form an enantiomer of Formula (VI). This may proceed via the use of an enzyme such as Novozym 435. Novozym 435 is a commercially available immobilised enzyme, specifically a lipase from Candida antarctica (CAL-B) attached to a porous acrylic resin (Lewatit VP OC 1600), used as a biocatalyst for reactions like esterification and transesterification.Accordingly, in some embodiments, the compound of Formula (I) is optically enriched. For example, a racemic mixture may be enriched by irradiation with left or right circularly polarized light, which when differentially exciting one enantiomer over the other, leads to selective photoisomerization of one isomer. Alternatively, chiral chromatography may be performed. This step may be performed as a purification step aimed at isolating a particular stereoisomer.

[0359] In some embodiments, the compound of Formula (I) is optically enriched with its R-enantiomer. In some embodiments, the compound of Formula (I) is optically enriched with its S-enantiomer. In some embodiments, the compound of Formula (I) is a R-enantiomer as represented by Formula (1-1):

[0360]

[0361] In some embodiments, the compound of Formula (I) is a S-enantiomer as represented by Formula (1-2):

[0362]

[0363] The present disclosure also relates to a compound of Formula (I), a salt, solvate, prodrug, stereoisomer or derivative thereof:

[0364]

[0365] wherein L is a linker selected from oxyacylene, aminoacylene, heteroarylene, oxylene, oxysulfonylene, oxyacyloxyene, and heterocyclylene;

[0366] i is an integer selected from 0 to 20;

[0367] m is an integer selected from 0 to 15; and

[0368] n is an integer selected from 0 to 15.

[0369] In some embodiments, the compound is a compound of Formula (I), or a salt, solvate, stereoisomer, prodrug, or derivative thereof:

[0370]

[0371] wherein

[0372] L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0373] i is an integer selected from 3 to 20;

[0374] m is an integer selected from 0 to 15; and

[0375] n is an integer selected from 0 to 15.

[0376] In some embodiments, the compound of Formula (I) is selected from:

[0377]

[0378] In some embodiments, i is an integer selected from 3 to 20, 5 to 20, 7 to 20, 10 to 20, 10 to 18, 10 to 16, or 10 to 15.

[0379] In some embodiments, m is an integer selected from 0 to 15, 1 to 15, 3 to 15, 5 to 15, 7 to 15, 9 to 15, 10 to 15, 10 to 14, 10 to 13 or 10 to 12. In some embodiments, m is 10.

[0380] In some embodiments, n is an integer selected from 0 to 15, 1 to 15, 1 to 13, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 2 to 5, or 3 to 5. In some embodiments, n is 5.

[0381] In some embodiments.i is an integer selected from 3 to 20;

[0382] m is an integer selected from 0 to 15; and

[0383] n is an integer selected from 0 to 15.

[0384] In some embodiments,

[0385] i is an integer selected from 10 to 16;

[0386] m is an integer selected from 10 to 14; and

[0387] n is an integer selected from 1 to 6.

[0388] In some embodiments,

[0389] i is an integer selected from 10 to 15;

[0390] m is an integer selected from 10 to 12; and

[0391] n is an integer selected from 4 to 6.

[0392] In some embodiments, L is aminoacylene, i is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, i is 15, m is 10 and n is 5. This corresponds to 12-SAASA. In some embodiments, i is 11, m is 10 and n is 5. This corresponds to 12-TAASA.

[0393] In some embodiments, L is triazolylene, i is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, i is 14, m is 10 and n is 5. This corresponds to 12-HDTZSA.

[0394] In some embodiments, L is oxylene, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0395] In some embodiments, L is oxysulfonylene, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0396] In some embodiments, L is oxyacyloxyene, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0397] In some embodiments, the compound of Formula (I) is deuterated. In some embodiments, the compound of Formula (I) is deuterated at at least one H position. In some embodiments, the compound of Formula (I) is deuterated at at least two, three, four or five H positions. In some embodiments, the compound of Formula (I) isdeuterated at all H positions. In some embodiments, the compound of Formula (I) is at an alpha position relative to L.

[0398] In some embodiments, the compound of Formula (I) is halogenated. In some embodiments, the compound is halogenated on its fatty acid ester chain. In some embodiments, the compound is halogenated on its hydroxy fatty acid backbone. In some embodiments, the compound is halogenated at one, two, three, four or five positions. In some embodiments, the compound is halogenated at all C positions on its fatty acid ester chain.

[0399] In some embodiments, the compound of Formula (I) is optically enriched. In some embodiments, the compound of Formula (I) is optically enriched with its R-enantiomer. In some embodiments, the compound of Formula (I) is optically enriched with its S-enantiomer. In some embodiments, the compound of Formula (I) is a R-enantiomer as represented by Formula (1-1):

[0400]

[0401] In some embodiments, the compound of Formula (I) is a S-enantiomer as represented by Formula (1-2):

[0402]

[0403] In some embodiments, the compound of Formula (lb) -(If) is characterised by resistance to acidic hydrolysis relative to a corresponding compound of Formula (la). In some embodiments, the acid resistance is about 5% to about 20%, about 5% to about 18%, about 5% to about 16%, about 5% to about 14%, or about 5% to about 12%.

[0404] The present disclosure also relates to a composition, comprising a compound of Formula (I), or a salt, solvate, prodrug, stereoisomer or derivative thereof, and optionally an excipient.

[0405] In some embodiments, the compound of Formula (I) is characterized by a concentration of about 10 mg / g to about 10,000 mg / g relative to the composition. In some embodiments, the concentration is about 50 mg / g to about 10,000 mg / g, about 100mg / g to about 10,000 mg / g, about 200 mg / g to about 10,000 mg / g, about 500 mg / g to about 10,000 mg / g, or about 1,000 mg / g to about 10,000 mg / g.

[0406] The present disclosure also relates to a method of treating a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof to the subject.

[0407] The present disclosure also relates to a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof for use in therapy.

[0408] The present disclosure also relates to a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof for use in treating a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption.

[0409] The present disclosure also relates to a use of a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof in the manufacture of a medicament for the treatment of a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption.

[0410] The present disclosure also relates to a method of remodeling a gut microbiome in a digestive system of a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof to the subject.

[0411] The present disclosure also relates to a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof for use in remodeling a gut microbiome in a digestive system.

[0412] The present disclosure also relates to a use of a compound of Formula (I), pharmaceutically acceptable salt, solvate, prodrug, derivative, stereoisomer or combination thereof in the manufacture of a medicament for the remodeling a gut microbiome in a digestive system.When L is oxyacylene, the compound is represented as a compound of Formula (la):

[0413]

[0414] wherein

[0415] i is an integer selected from 0 to 15;

[0416] m is an integer selected from 5 to 10; and

[0417] n is an integer selected from 5 to 10.

[0418] In some embodiments, i is an integer selected from 0 to 15, 5 to 15, 7 to 15, 8 to 15, 9 to 15, 10 to 15, or 11 to 15. In some embodiments, m is an integer selected from 5 to 10, 7 to 10, 8 to 10, or 9 to 10. In some embodiments, n is an integer selected from 5 to 9, 5 to 8, 5 to 7, or 5 to 6.

[0419] In some embodiments, when L is oxyacylene, i is 15, m is 8, and n is 7. This corresponds to 10-SAHSA.

[0420] In some embodiments, when L is oxyacylene, i is 3, m is 10, and n is 5. This corresponds to 12-HAHSA.

[0421] In some embodiments, when L is oxyacylene, i is 13, m is 10, and n is 5. This corresponds to 12-PAHSA.

[0422] In some embodiments, when L is oxyacylene, i is 15, m is 10, and n is 5. This corresponds to 12-SAHSA.

[0423] In some embodiments, when L is oxyacylene, i is 11, m is 10, and n is 5. This corresponds to 12-TAHSA.

[0424] In some embodiments,

[0425] when L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;

[0426] i is an integer selected from 0 to 20;

[0427] m is an integer selected from 0 to 15; and

[0428] n is an integer selected from 0 to 15.

[0429] In some embodiments, i is an integer selected from 3 to 20, 5 to 20, 7 to 20, 10 to 20,10 to 18, 10 to 16, or 10 to 15.

[0430] In some embodiments, m is an integer selected from 0 to 15, 1 to 15, 3 to 15, 5 to 15, 7 to 15, 9 to 15, 10 to 15, 10 to 14, 10 to 13 or 10 to 12. In some embodiments, m is 10.

[0431] In some embodiments, n is an integer selected from 0 to 15, 1 to 15, 1 to 13, 1 to 10, 1 to 8, 1 to 6, 1 to 5, 2 to 5, or 3 to 5. In some embodiments, n is 5.

[0432] In some embodiments,

[0433] i is an integer selected from 3 to 20;

[0434] m is an integer selected from 0 to 15; and

[0435] n is an integer selected from 0 to 15.

[0436] In some embodiments,

[0437] i is an integer selected from 10 to 16;

[0438] m is an integer selected from 10 to 14; and

[0439] n is an integer selected from 1 to 6.

[0440] When L is aminoacylene, the compound is represented as a compound of Formula (lb):

[0441] O

[0442]

[0443] In some embodiments, i is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, i is 15, m is 10 and n is 5. This corresponds to 12-SAASA. In some embodiments, i is 11, m is 10 and n is 5. This corresponds to 12-TAASA.

[0444] When L is heteroarylene, the heteroarylene is selected from triazolylene. In some embodiments, the compound is a compound of Formula (Ic):

[0445]

[0446] In some embodiments, i is an integer selected from 10 to 16, m is 10 and n is 5. In some embodiments, i is 14, m is 10 and n is 5. This corresponds to 12-HDTZSA.When L is oxylene, the compound is represented as a compound of Formula (Id):

[0447]

[0448] (Id)

[0449] In some embodiments, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0450] When L is oxysulfonylene, the compound is represented as a compound of Formula (le):

[0451]

[0452] In some embodiments, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0453] When L is oxyacyloxyene, the compound is represented as a compound of Formula (If):

[0454] O

[0455] "^^0^0 O

[0456]

[0457] (If).

[0458] In some embodiments, i is an integer selected from 10 to 16, m is 10 and n is 5.

[0459] It was found that CIO- and C12- branched FAHFA regioisomers outperformed other positions, implying that their 3D presentation favors interaction with lipid-processing molecular targets. These CIO and C12 compounds were also largely insulin-insensitive.

[0460] In some embodiments, the disease or condition associated with lipid accumulation is selected from a metabolic disease and / or an inflammatory disease. In some embodiments, the disease or condition associated with lipid accumulation is selected from dyslipidemia, hypertriglyceridemia, mixed dyslipidemia, obesity, insulin resistance, impaired glucose tolerance, type 2 diabetes, metabolic syndrome, visceral adiposity, fatty liver disease including metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), alcohol-associated fatty liver disease, steatotic liver with fibrosis, liver cirrhosis secondary to steatosis, pancreatic steatosis, skeletal muscle steatosis, cardiac steatosis, atheroscleroticcardiovascular disease, hyperlipidemia-associated hypertension, imbalanced gut microbiome, increased intestinal permeability, inflammatory bowel diseases including ulcerative colitis and Crohn's disease, microbiome-associated inflammatory conditions, chronic low-grade inflammation associated with ectopic lipid deposition, and inflammatory diseases associated with dyslipidemia and metabolic dysfunction. The disease or condition associated with lipid accumulation may be a multifactorial disease, which arises from multiple interacting causes or influences, rather than a single process, sign, or symptom. For example, the multifactorial disease may be caused by at least two of metabolic syndrome, obesity, dyslipidemia, insulin resistance, and gut health.

[0461] In some embodiments, the compound of Formula (I) inhibits lipid absorption.

[0462] In some embodiments, the method or use is independent on the presence of insulin. In some embodiments, the method or use is characterised by a substantially similar lipid-lowering efficacy with or without insulin. In some embodiments, the method or use is characterised by a tolerance to insulin.

[0463] In some embodiments, the method or use is independent on a presence of glucose. In some embodiments, the method or use is independent on an uptake of glucose. In some embodiments, the method or use is characterised by a substantially similar lipid-lowering efficacy with or without glucose. In some embodiments, the method or use does not inhibit or impair glucose uptake or absorption. In some embodiments, the method or use is characterised by a tolerance to glucose uptake or absorption. The method or use may thus be suitable for treating a subject with obesity, insulin resistance, or fatty liver disease even when those subjects are on glucose-lowering medications (such as GLP-lra and insulin), since the compound of Formula (I) does not exacerbate low blood glucose.

[0464] In some embodiments, the method or use enriches short-chain fatty acid (SCFA) producing bacteria in a gut or intestine of the subject. The SCFA may be acetate, propionate, and butyrate. In some embodiments, the method or use selectively promotes growth of SCFA producing bacteria in a gut or intestine of the subject. The SCFA producing bacteria may be Blautia, Lachnospiraceae NK4A136, and Roseburia.

[0465] In some embodiments, the method or use is characterised by a reduced ectopic fat deposition. The ectopic fat deposition may be in a liver and / or visceral fat adipose tissue of the subject.In some embodiments, the method or use comprises orally administrating the compound of Formula (I).

[0466] In some embodiments, the compound of Formula (I) is administered at a dose of about 50 mg / kg to about 1500 mg / kg. In some embodiments, the compound of Formula (I) is administered at a dose of about 50 mg / kg to about 1400 mg / kg, about 50 mg / kg to about 1300 mg / kg, about 50 mg / kg to about 1200 mg / kg, about 50 mg / kg to about 1100 mg / kg, about 50 mg / kg to about 1000 mg / kg, about 50 mg / kg to about 900 mg / kg, or about 50 mg / kg to about 800 mg / kg.

[0467] The compound of Formula (I) may be administered to the subject at a dosage of about 200 mg / day. This may be via fecal-throughput scaling, which provides the safest, and most practical way. The compound of Formula (I) may be delivered to the subject at a dosage of about 200 mg / day to about 800 mg / day, about 300 mg / day to about 800 mg / day, about 400 mg / day to about 800 mg / day, about 500 mg / day to about 800 mg / day, or about 500 mg / day to about 700 mg / day. This offers a pragmatic midpoint, balancing luminal & epithelial coverage. It is expected that a high probability of efficacy, practical, justifiable safety profile will be achieved. The compound of Formula (I) may be delivered to the subject at a dosage of about 800 mg / day to about 2000 mg / day, about 900 mg / day to about 2000 mg / day, about 1000 mg / day to about 2000 mg / day, about 1000 mg / day to about 1800 mg / day, or about 1000 mg / day to about 1500 mg / day. This may offer full surface-area coverage and the highest efficacy assurance.

[0468] The compound can be administered to a subject as a pharmaceutically acceptable salt thereof. Suitable pharmaceutically acceptable salts include, but are not limited to salts of pharmaceutically acceptable inorganic acids such as hydrochloric, sulphuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, maleic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, toluenesulphonic, benezenesulphonic, salicyclic sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids.

[0469] Base salts include, but are not limited to, those formed with pharmaceutically acceptable cations, such as sodium, potassium, lithium, calcium, magnesium, ammonium and alkylammonium. In particular, the present invention includes within its scope cationicsalts, e.g. sodium or potassium salts, or alkyl esters (e.g. methyl, ethyl) of the carboxylic acid group.

[0470] Basic nitrogen-containing groups may be quarternised with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides and iodides; dialkyl sulfates like dimethyl and diethyl sulfates; and others.

[0471] It will be appreciated that any compound that is a prodrug of the compound of formula (I) is also within the scope and spirit of the invention. Thus the compound can be administered to a subject in the form of a pharmaceutically acceptable pro-drug. The term "pro-drug" is used in its broadest sense and encompasses those derivatives that are converted in vivo to the compound of the invention. Such derivatives would readily occur to those skilled in the art. Other texts which generally describe prodrugs (and the preparation thereof) include: Design of Prodrugs, 1985, H. Bundgaard (Elsevier); The Practice of Medicinal Chemistry, 1996, Camille G. Wermuth et al., Chapter 31 (Academic Press); and A Textbook of Drug Design and Development, 1991, Bundgaard et al., Chapter 5, (Harwood Academic Publishers). For example, the carboxylic acid moiety may be protected through esterification.

[0472] The compound may be in crystalline form either as the free compound or as a solvate (e.g. hydrate) and it is intended that both forms are within the scope of the present disclosure. Methods of solvation are generally known within the art.

[0473] The compound of the disclosure, or a pharmaceutically acceptable salt, solvate stereoisomer, prodrug or derivative thereof is administered to the patient in a therapeutically effective amount. As used herein, a therapeutically effective amount is intended to include at least partially attaining the desired effect, or delaying the onset of, or inhibiting the progression of, or halting or reversing altogether the onset or progression of macular degeneration.

[0474] The term "therapeutic effect" refers to some extent of relief of one or more of the symptoms of a disorder (e.g., a neoplasia or tumor) or its associated pathology. " Therapeutically effective amount" as used herein refers to an amount of an agent which is effective, upon single or multiple dose administration to the cell or subject, in prolonging the survivability of the patient with such a disorder, reducing one or more signs or symptoms of the disorder, preventing or delaying, and the like beyond that expected in the absence of such treatment. " Therapeutically effective amount" isintended to qualify the amount required to achieve a therapeutic effect. A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the "therapeutically effective amount" (e.g., ED50) of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in a pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0475] As used herein, the term "effective amount" relates to an amount of compound which, when administered according to a desired dosing regimen, provides the desired therapeutic activity. Dosing may occur at intervals of minutes, hours, days, weeks, months or years or continuously over any one of these periods. Suitable dosages may lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. In one embodiment, the dosage may be in the range of 1 mg to 500 mg per kg of body weight per dosage. In another embodiment, the dosage may be in the range of 1 mg to 250 mg per kg of body weight per dosage. In yet another embodiment, the dosage may be in the range of 1 mg to 100 mg per kg of body weight per dosage, such as up to 50 mg per body weight per dosage.

[0476] Suitable dosage amounts and dosing regimens can be determined by the attending physician and may depend on the severity of the condition as well as the general age, health and weight of the patient to be treated.

[0477] The compound of the disclosure may be administered in a single dose or a series of doses. While it is possible for the active ingredient to be administered alone, it is preferable to present it as a composition, preferably as a pharmaceutical composition. The formulation of such compositions is well known to those skilled in the art. The composition may contain any suitable carriers, diluents or excipients. These include all conventional solvents, dispersion media, fillers, solid carriers, coatings, antifungal and antibacterial agents, dermal penetration agents, surfactants, isotonic and absorption agents and the like. It will be understood that the compositions of the invention may also include other supplementary physiologically active agents.

[0478] The carrier must be pharmaceutically "acceptable" in the sense of being compatible with the other ingredients of the composition and not injurious to the patient. The compositions may conveniently be presented in unit dosage form and may be preparedby any methods well known in the art of pharmacy. Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.

[0479] The compound or composition of the disclosure may be suitable for intravenous administration. For example, a compound of formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug or derivative thereof may be administered intravenously at a dose of up to 16 mg / m2.

[0480] The compound or composition of the disclosure may also be suitable for oral administration and may be presented as discrete units such as capsules, sachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste. In another embodiment, the compound of formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug or derivative is orally administerable.

[0481] A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder (e.g. inert diluent, preservative disintegrant (e.g. sodium starch glycolate, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose) surface-active or dispersing agent. Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

[0482] The compound or composition of the disclosure may be suitable for topical administration in the mouth including lozenges comprising the active ingredient in a flavoured base, usually sucrose and acacia or tragacanth gum; pastilles comprising theactive ingredient in an inert basis such as gelatine and glycerin, or sucrose and acacia gum; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0483] The compound or composition of the disclosure may be suitable for topical administration to the skin may comprise the compounds dissolved or suspended in any suitable carrier or base and may be in the form of lotions, gel, creams, pastes, ointments and the like. Suitable carriers include mineral oil, propylene glycol, polyoxyethylene, polyoxypropylene, emulsifying wax, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Transdermal patches may also be used to administer the compounds of the invention.

[0484] The compound or composition of the disclosure may be suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain anti-oxidants, buffers, bactericides and solutes which render the compound, composition or combination isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compound, composition or combination may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

[0485] Preferred unit dosage composition or combinations are those containing a daily dose or unit, daily sub-dose, as herein above described, or an appropriate fraction thereof, of the active ingredient.

[0486] It should be understood that in addition to the active ingredients particularly mentioned above, the composition or combination of this invention may include other agents conventional in the art having regard to the type of composition or combination in question, for example, those suitable for oral administration may include such further agents as binders, sweeteners, thickeners, flavouring agents disintegrating agents, coating agents, preservatives, lubricants and / or time delay agents. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharine. Suitable disintegrating agents include cornstarch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable flavouring agents include peppermint oil, oil of Wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents includepolymers or copolymers of acrylic acid and / or methacrylic acid and / or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate.

[0487] Examples

[0488] Scalable synthetic platform yields potent FAHFAs analogues

[0489] Recognizing the substantial potential of FAHFAs, the inventors designed efficient and scalable synthetic methods to reliably produce the entire FAHFA family for their commercial and biomedical applications (Figure 2A). The synthesis of each analog starts with the selective addition of Grignard reagents to an aldehyde, producing various hydroxy-fatty acids 2 in good quantities (Figure 2A, step a). These hydroxy-fatty acids are then refluxed in methanol with thionyl chloride to form hydroxy-fatty acid methyl esters 3 (Figure 2A, step b). Esterification of the secondary alcohol with the appropriate acid chloride and pyridine leads to the formation of FAHFA methyl esters 4 (Figure 2A, step c). Finally, selective hydrolysis of the more accessible methyl ester yields compound 5 in good overall yield (Figure 2A, step d).

[0490] Using this streamlined four-step sequence, the inventors successfully synthesized 49 members of the FAHFA family and 15 derivatives spanning various chain lengths and regiochemistries with high regioselectivity and purity, meticulously adhering to established naming conventions (Figure 2B). Each member has been carefully named and characterized through a systematic approach, facilitating the distinction and identification of the compounds.

[0491] High lipid absorption from daily intake can lead to excessive fat accumulation in the body, resulting in weight gain and adiposity. To investigate the potential anti-obesity effects of synthetic FAHFAs, a screening was conducted to assess their inhibitory effect on cellular lipid absorption in HepG2 cells, a commonly used model for metabolism and toxicity studies. The screening involved varying concentrations of FAHFAs (1-100 pg / mL) under two conditions, with and without insulin stimulation. The results were normalized to DMSO-treated cells to determine the Iog2 fold change in lipid absorption rate. Disulfiram-treated cells were used as baseline and a p-value <0.1 and Iog2 fold change <-0.8 were identified as the thresholds to shortlist potential FAHFAs (Figure 3A). The screen revealed that 10 of the FAHFA analogues significantly suppressed lipiduptake under insulin-stimulated conditions, and 6 were effective under basal conditions; notably, 4 FAHFAs inhibited lipid uptake in both contexts. Based on the screening, 12 FAHFAs were shortlisted for their inhibitory effect on lipid absorption. These 12 FAHFAs are 9-HAHSA, 10-HAHSA, 10-PAHSA, 10-SAHSA, 12-HAHSA, 12-PAHSA, 12-SAHSA, 12-TAHSA, 12-PAHIA, 17-SAHSA, 17-HAHSA and 17-PAHSA. The shortlisted FAHFAs are presented in a Venn diagram (Figure 3B).

[0492] A dose-response study was performed to further evaluate of the efficacy and toxicity of the shortlisted FAHFAs at a wider concentration range (i.e. 1.56-200 pg / mL). Most FAHFAs did not obey a typical sigmoidal dose response in their inhibition on lipid uptake but instead exhibited a non-monotonic (U-shaped) dose response with poor inhibitory efficacy at very low and very high concentrations (Figure 4A), likely due to solubility limits at higher doses given their hydrophobic nature. Five of the shortlisted FAHFAs, i.e. 10-HAHSA, 12-HAHSA, 12-PAHSA, 12-SAHSA and 12-TAHSA, demonstrated comparatively wider range of effective inhibitory concentrations and desirable inhibition (>40%) compared to DMSO-treated cells. The presence of insulin did not affect their lipid-lowering activity. Among them, 12-SAHSA and 12-TAHSA were non-cytotoxic (or had minimal cytotoxicity) even at high concentrations and could potentially enhance glucose uptake (Figure 4B-C). Given the metabolic benefits and favorable safety profiles, 12-SAHSA and 12-TAHSA were chosen for subsequent chemical modifications and preclinical testing.

[0493] Bioisoteric bond modifications enhance stability and selectivity

[0494] To further potentiate the therapeutic efficacy, and to overcome the inherent instability of the ester bond which is susceptible to degradation by acid hydrolysis and thus negatively impacting its chemical stability during oral administration, 15 derivatives of 12-SAHSA and 12-TAHSA bearing different chemical modifications and ester bond bioisosteres were synthesized. Replacement of the ester bonds linking the hydroxyl group of one fatty acid and the carboxyl group of another fatty acid with other acid-tolerant functional groups, such as carbonate ester (12-DCAHSA and 12-HCAHSA), sulfonate ester (12-DSAHSA and 12-HSAHSA), ether (12-TOHSA and 12-OOHSA), and amide (12-TAASA and 12-SAASA), should improve the structural integrity of FAHFAs, thus facilitating oral delivery and in vivo uptake of FAHFAs (Figure 2B). The ester group with known isosteres were also synthesized, such as 1,2,3-triazoles (Figure 2B; 12-DDTZSA and 12-HDTZSA). Additionally, other chemical modifications to 12-SAHSA and 12-TAHSA were also introduced, such as the inclusion of heavy isotopes like deuterium (12-d-TAHSA and 12-d-SAHSA) to induce a kinetic isotope effect and changing thechirality of the FAHFAs, aiming to slow ester hydrolysis. Enantioenriched FAHFAs ((R)-12-TAHSA and (R)-12-SAHSA) and fluorinated FAHFA (12-PFTeDAHSA) were prepared to probe other aspects of stability and activity.

[0495] Certain modifications conferred improved chemical stability under harsh gastric conditions (pH 1-2), as evidenced by in vitro stability assays (Figure 15). Notably, 12-TAASA and 12-HDTZSA were 12.64% and 9.20% more resistant to acidic hydrolysis than the parental 12-SAHSA, respectively, significantly enhancing their suitability for oral administration.

[0496] The suppressive effect of these synthetic FAHFA derivatives on lipid uptake was validated in two hepatic cell lines, HepG2 (Figure 5) and AML12 (Figure 6). Similar to their parental structures, most 12-SAHSA and 12-TAHSA derivatives effectively inhibited lipid absorption in different hepatocyte cell lines at a concentration of 6.25 to 100 pg / mL. The inhibitory effect was more evident in HepG2 compared to AML12, likely due to the high metabolic rate of the hepatocellular carcinoma line. The therapeutic ranges also showed subtle variability between different FAHFAs derivatives, highlighting the influence of functional groups and chemical properties on their lipid-lowering ability.

[0497] Unlike their parental structures, certain 12-SAHSA and 12-TAHSA derivatives showed a suppressive effect on glucose uptake at a high concentration (200 pg / mL) (Figure 7 & 8). d-12-TAHSA had the strongest inhibition on glucose uptake in both HepG2 (-18.5%) and AML12 (-29.1%) while 12-SAASA (an amide analog of 12-SAHSA), 12-TAASA (an amide analog of 12-TAHSA), and 12-HDTZSA (a triazole analog of 12-HSAHSA) had the least influence on glucose absorption. These derivatives exhibited potent inhibition of lipid uptake with minimal cytotoxicity and, crucially, retained high specificity for lipid absorption (showing little or no inhibition of glucose uptake). 12-SAASA, 12-TAASA and 12-HDTZSA were also non-cytotoxic to the hepatocytes (Figure 9 & 10).

[0498] All bioisosteric modifications conferred at least some improvement in chemical stability under harsh gastric conditions (pH 1-2), as evidenced by in vitro stability assays. Notably, 12-TAASA and 12-HDTZSA were 12.64% and 9.20% more resistant to acidic hydrolysis than the parental 12-SAHSA, respectively, significantly enhancing their suitability for oral administration. Some other derivatives were not advanced due to various issues: for instance, certain deuterated and R-stereoisomers had only narrow effective concentration ranges for lipid inhibition, some carbonate / sulfonate analogues (e.g., 12-DCAHSA, 12-DSAHSA) unexpectedly impaired glucose uptake, and a fewcompounds showed toxicity at higher doses. These outcomes highlight how the bioactivity of synthetic FAHFAs is sensitive to functional group alterations and physicochemical properties.

[0499] Analysis highlighted 12-SAASA, 12-TAASA and 12-HDTZSA as candidates for further development due to their potent lipid-lowering capacity and minimal impact on glucose absorption and favorable cytotoxicity profile. These derivatives also demonstrated improved chemical stability, hence suitable for weight control via oral administration.

[0500] In vivo anti-obesity efficacy comparable to semaglutide

[0501] To examine the in vivo weight loss efficacy, a liver disease progression aggravation diet (LIDPAD) mouse model was utilised, in which C57BL / 6J mice were fed an optimized high-fat, high-calorie diet that accelerated metabolic dysfunction-associated steatotic liver disease (MASLD) progression. After diet-induced obesity was established, mice were treated once daily by oral gavage dosing of 100 mg / kg 12-SAHSA, 12-TAHSA and their derivatives, 12-SAASA, 12-TAASA and 12-HDTZSA for a month. A group of obese mice treated with the FDA-approved anti-obesity drug semaglutide (referred to as "sema") served as a benchmark for efficacy, and untreated obese mice on LIDPAD served as control.

[0502] The suppressive effect of 12-SAHSA, 12-TAASA and 12-HDTZSA on weight gain caused by the obesogenic diet was on par with semaglutide, an FDA-approved weight loss drug (Figure 11A). Mice receiving 12-TAASA or 12-HDTZSA over the month-long treatment gained less weight than untreated controls, despite the obesogenic diet. These three FAHFA derivatives also improved the glucose tolerance and substantially reduced ectopic fat deposition in the liver and visceral fat adipose tissues, clearly highlighting their beneficial metabolic effects (Figure 11B-D). Thus, oral FAHFA therapy delivered metabolic benefits on par with a potent injectable drug.

[0503] In addition to slowing weight gain, FAHFA treatment markedly diminished ectopic fat deposition in key metabolic tissues. Mice given 12-TAASA, 12-HDTZSA, or 12-SAHSA had notably lower liver and visceral white adipose tissue (WAT) weights than untreated obese mice, indicating reduced hepatic steatosis and visceral fat accumulation. Histological analysis confirmed that FAHFA-treated livers and adipose depots were protected from the severe lipid engorgement seen in controls. Even the parent FAHFAs (12-TAHSA and 12-SAASA), though slightly less effective in curbing weight gain, still conferred significant reductions in visceral fat accumulation and improved insulinsensitivity. Importantly, no adverse effects were observed: the organ weights of heart, spleen and kidney were largely unaffected by most FAHFA derivatives, which suggests the high dose was well-tolerated by the mice with no significant tissue injury (Figure 12A).

[0504] Gut microbiome remodeling and SCFA enrichment by synthetic FAHFAs

[0505] To investigate the mechanisms underlying these metabolic improvements, the fate and distribution of FAHFAs in vivo was considered. After a single oral dose (100 mg / kg), both parent and synthetic FAHFAs were detected in fecal samples, but none in plasma (data not shown), suggesting minimal systemic absorption. Intriguingly, the pharmacokinetic profiles of synthetic vs. natural FAHFAs differed: the levels of 12-SAASA, 12-TAASA, and 12-HDTZSA in feces peaked at ~24 h post-gavage and returned to baseline by 48 h, whereas the parent 12-SAHSA and 12-TAHSA remained elevated even at 48 h (Figure 13A). This indicates that the engineered derivatives might undergo more complete metabolism or uptake in the gut, resulting in less fecal excretion. It is hypothesized that gut microbiota or intestinal enzymes could be processing the synthetic FAHFAs differently, potentially contributing to their enhanced efficacy.

[0506] 16S rRNA gene sequencing was performed on the gut microbiota of mice after four weeks of treatment. Microbiome analysis revealed pronounced shifts in community composition in response to the synthetic FAHFAs. Principal coordinates analysis of betadiversity showed that mice treated with 12-TAASA or 12-HDTZSA formed distinct, tight clusters separate from untreated obese controls, indicating a consistent remodeling of the microbiota (Figure 13B). In contrast, mice given the parent FAHFAs had microbiome profiles overlapping with controls, suggesting minimal impact on the microbiota. At the genus level, 12-TAASA and 12-HDTZSA treatment led to a significant depletion of Romboutsia and Olsenella (genera associated with inflammation in high-fat diets) alongside an enrichment of Blautia, a genus of known SCFA-producing bacteria, compared to the LIDPAD control group (Figure 13C). 12-SAASA induced a similar increase in Blautia and decrease in Olsenella, though it was less effective at reducing Romboutsia. These compositional changes suggest that synthetic FAHFAs create a gut environment favoring beneficial microbes over potentially deleterious ones. Notably, such changes were not observed with semaglutide in this period (consistent with GLP-1 RA primarily acting on host appetite and not directly on gut flora).

[0507] Functionally, the microbiomes of 12-TAASA– and 12-HDTZSA–treated mice were enriched for pathways involved in short-chain fatty acid (SCFA) production.Metagenomic inferences pointed to increased microbial fermentation of dietary fibers and glycolytic intermediates (e.g. pyruvate, lactate) into SCFAs like acetate, propionate, and butyrate (Figure 4D). Correspondingly, these treatments selectively promoted the growth of key SCFA-producing taxa including Blautia, Lachnospiraceae NK4A136, and Roseburia (Figure 13E). Such bacteria are well-known for their beneficial metabolic effects, as SCFAs they produce can improve host energy balance, gut barrier integrity, and anti-inflammatory status. Taken together, these data demonstrate that synthetic FAHFA derivatives remodel the gut microbiota, expanding SCFA-producing populations while suppressing pro-inflammatory microbes. This microbiome modulation is likely an important contributor to the observed metabolic benefits, consistent with literature linking SCFA-producing microbiota to protection against obesity and metabolic dysbiosis.

[0508] FAHFA 12-TAASA directly suppresses intestinal lipid absorption pathways

[0509] Beyond microbiome effects, the direct actions of FAHFAs on the host intestinal epithelium was also explored. Focus was on 12-TAASA, the top-performing compound, in light of its potent lipid-lowering activity in vivo. Transcriptomic profiling (RNA-seq) of small intestinal mucosa from treated mice revealed a distinct gene expression signature induced by 12-TAASA compared to other groups (Figure 14A). All FAHFA treatments appeared to stimulate genes involved in cell proliferation in the intestine, suggesting a general trophic effect on the mucosa (potentially aiding epithelial turnover and gut barrier maintenance). However, 12-TAASA (and to a lesser extent its parent 12-SAHSA) uniquely downregulated genes related to fibrogenesis and lipid metabolism (Figure 14B). Specifically, pathway analysis showed that 12-SAHSA mainly suppressed adipokine signaling pathways, whereas 12-TAASA broadly repressed the expression of genes involved in the transport and processing of dietary lipids (cholesterol, fatty acids, ketone bodies, steroids) (Figure 14C). Other FAHFAs had distinct transcriptomic effects, e.g., 12-TAHSA and 12-SAASA upregulated amino acid metabolism genes, and 12-HDTZSA modulated immune response genes, but these were not directly related to lipid absorption. The intestinal gene expression profile of 12-TAASA thus pointed to a potent local inhibition of lipid uptake pathways, consistent with its ability to prevent diet-induced fat accumulation.

[0510] Key molecular targets were identified through which 12-TAASA might be mediating this effect. Using a protein-protein interaction network analysis of the genes most strongly downregulated by 12-TAASA, several "hub" genes were identified that could serve as central regulators of intestinal lipid absorption. Top hub candidates included Cd44,Aldh1a1, Apoa1, Ugt1a1, Cyp2d26, Hspa1b, Hmgcs2, Fabp2, Dgat2 and Tlr2 (Figure 14D). Strikingly, five of these (Cd44, Apoa1, Fabp2, Dgat2, Tlr2) encode cell-surface or membrane-associated proteins, raising the possibility that 12-TAASA might directly bind to and inhibit these targets on intestinal cells.

[0511] To test if 12-TAASA physically interacts with intestinal receptors or transporters, a biotinylated 12-TAASA probe was synthesized and ligand pull-down assays were performed on intestinal mucosal lysates (Figure 14E). Proteomic analysis of pulled-down proteins identified 468 putative 12-TAASA-binding partners, including 134 membrane proteins. Many of these interactors are involved in lipid metabolism, transport, or endocytosis (Figure 14F), aligning well with the pathways predicted to be suppressed by 12-TAASA. Notably, CD44, one of the top hub genes from the transcriptomic analysis, was confirmed as a direct binding target of 12-TAASA in the pull-down (Figure 14G). CD44 is a cell-surface glycoprotein implicated in fatty acid translocase activity and inflammatory signaling in the gut; its interaction with 12-TAASA suggests a mechanism for blocking fat absorption and associated inflammation. Two additional overlapping targets (Chp1 and Hspa1 family proteins) were identified in both the RNA-seq and proteomics datasets, further supporting a convergence of computational and experimental evidence for specific host targets of 12-TAASA. The membrane proteins bound by 12-TAASA formed a dense interaction network with each other and with genes downregulated by 12-TAASA, highlighting a coordinated mode of action. Together, these results provide compelling evidence that 12-TAASA directly interacts with key intestinal receptors / transporters (such as CD44) to inhibit dietary lipid uptake, thereby acting as a "molecular brake" on fat absorption in the gut. This dual mechanism— where 12-TAASA both reshapes the microbiome and directly targets host pathways— underpins its robust metabolic benefits in obese mice.

[0512] Discussion

[0513] This study presents advances in the chemistry and biology of FAHFAs, addressing longstanding obstacles to their development as therapeutics. A modular, scalable synthesis platform was established, capable of producing over 60 FAHFA analogues and derivatives, encompassing both naturally occurring isomers and new structural variants. This broad repertoire was critical for comprehensive structure-activity relationship analysis. Two innovations underline our approach: (1) an optimized Grignard-based stereoselective route to diverse hydroxy-fatty acid precursors, and (2) the strategic bioisosteric replacement of the metabolically labile ester bond with an amide or 1,2,3-triazole linkage. These modifications dramatically enhanced resistance to enzymatic andacidic degradation while improving oral bioavailability, effectively overcoming the pharmacokinetic limitations of native FAHFAs. In effect, the molecular framework of FAHFAs was "redesigned" to be drug-like, without sacrificing their beneficial bioactivity.

[0514] Among the synthesized analogues, 12-TAASA emerged as a lead candidate with remarkable multimodal efficacy. Four weeks of oral 12-TAASA therapy in obese mice led to significant weight reduction, suppression of hepatic and visceral fat accumulation, and improved glucose tolerance and insulin sensitivity. These outcomes are particularly noteworthy given that the effect of 12-TAASA in this diet model were comparable to those of semaglutide, one of the most potent anti-obesity drugs currently available. An orally active small molecule matching the efficacy of an injectable peptide is a rare achievement in this field.

[0515] Mechanistically, this work sheds new light on how FAHFA derivatives can exert metabolic benefits through dual pathways involving both host and microbiome. The pronounced gut microbiota remodeling observed with 12-TAASA and 12-HDTZSA treatment addresses a gap in the literature, as the influence of FAHFAs on the gut microbiome had previously been poorly defined. These synthetic FAHFAs consistently enriched SCFA-producing genera (e.g., Blautia, Roseburia) and depleted potentially harmful taxa (e.g., Romboutsia). SCFAs like acetate and butyrate have well-documented roles in enhancing metabolic health, including improving insulin sensitivity, inducing satiety, and fortifying the gut barrier. Thus, the microbiome changes induced by FAHFA therapy likely contribute substantially to its anti-obesogenic effect, in line with emerging concepts of treating metabolic disease by modulating the gut microbiota. It is intriguing to speculate that native FAHFAs may also signal through gut microbes; indeed, prior studies suggested that 5- and 9-PAHSA can activate GPR120 on intestinal cells to promote antimicrobial peptide release, thereby indirectly shaping microbial communities. By introducing amide bonds and other modifications, 12-TAASA appears to amplify these microbiome-mediated effects, by prolonging FAHFA presence in the distal gut where most microbes reside. Enhanced SCFA production in 12-TAASA-treated mice provide a plausible link between the microbiota shifts and the observed improvements in adiposity and glycemic control.

[0516] Concurrently, 12-TAASA directly targets the host's intestinal nutrient handling. Transcriptomic and proteomic analyses converged on CD44 as a key mediator of 12-TAASA's action in the gut. CD44 has been implicated in fatty acid uptake and cholesterol metabolism in enterocytes. By binding to CD44 (and possibly other surface proteins likeApoA1 and FABP2), 12-TAASA may interfere with the normal absorption of dietary fats. This mechanism is somewhat reminiscent of intestinal lipase inhibitors (like orlistat) or bile acid sequestrants, which also reduce lipid absorption, but 12-TAASA achieves this through a novel receptor-mediated route rather than broad enzymatic blockade. The result is a more targeted "braking" of lipid uptake, as evidenced by reduced expression of lipid transport genes and lower fat deposition in 12-TAASA-treated animals. Notably, this dual mechanism-of-action (microbiome + host receptors) is unique among obesity drugs. Current medications typically act either on the host endocrine system (e.g., GLP-1 analogues on appetite and insulin) or on nutrient absorption (e.g., orlistat on pancreatic lipase) but do not intentionally modulate the gut microbiota. 12-TAASA and its analogues, by affecting both domains, could represent a new class of "two-pronged" therapeutics for metabolic disease.

[0517] Beyond weight loss and lipid metabolism, these findings hint at broader health benefits of synthetic FAHFAs. The apparent stimulation of intestinal epithelial proliferation by FAHFAs may support mucosal healing and barrier function. This could make them useful in conditions like inflammatory bowel disease or "leaky gut" syndrome, where enhancing epithelial regeneration and integrity is desirable. Additionally, FAHFAs have been reported by others to enhance insulin-stimulated glucose uptake in muscle and fat tissues, suggesting potential as insulin sensitizers or adjuncts in diabetes management. Because they are lipids, FAHFAs (especially with our modifications) are inherently membrane-permeable and could distribute to various tissues, possibly exerting systemic anti-inflammatory effects. Taken together, these properties position FAHFA derivatives as versatile therapeutic candidates that address multiple facets of metabolic syndrome, obesity, dyslipidemia, insulin resistance, and gut health.

[0518] Implications and Potential Applications

[0519] Naturally occurring (12-SAHSA & 12-TAHSA) and modified (notably 12-TAASA and 12-HDTZSA) FAHFAs were identified that potently inhibit cellular lipid uptake without compromising glucose absorption. These FAHFA candidates demonstrated good safety profile and tolerability in preclinical mouse model. In a diet-induced obesity mouse model, oral administration of these derivatives reduced weight gain, improved glucose tolerance, and attenuated ectopic lipid deposition, achieving efficacy comparable to the GLP-1 agonist Semaglutide. Mechanistically, the 12-TAASA and 12-HDTZSA act through a dual mechanism: remodeling the gut microbiome to enrich SCFA-producing, taxa and directly suppressing intestinal lipid absorption via ligand-receptor interactions and downregulation of key transport genes. These findings establish synthetic FAHFAs aspromising class of dual-action therapeutics targeting both host lipid metabolism and gut microbiota, offering a new strategy against obesity and metabolic disorders.

[0520] Conclusion

[0521] In summary, this work represents a significant step forward in lipid-based therapeutics for metabolic disease. By combining cutting-edge chemical synthesis with innovative biological insight, FAHFA molecules were engineered for enhanced stability and potency and demonstrated their dual-action efficacy in an obesity model. 12-TAASA exemplifies this new class of compounds, acting simultaneously on the gut microbiome and intestinal lipid absorption to produce metabolic benefits. This convergence of chemistry and microbiology opens an entirely new avenue for treating obesity and related disorders. Synthetic FAHFAs can evolve into a novel oral therapy that safely mimics the metabolic advantages of bariatric surgery (which also alters gut hormones and microbiota) in a pill-based form. The translational potential of these findings is high. Ultimately, FAHFA-based therapeutics offer a promising strategy to combat obesity, type 2 diabetes, and even inflammatory diseases by holistically targeting the interconnected networks of host metabolism and the gut ecosystem.

[0522] Regioisomer position determines lipid-lowering efficacy of FAHFAs for potential MASLD therapeutics

[0523] To nominate FAHFA chemotypes that could inhibit gut-to-liver lipid flux, an investigation was performed to understand which species most effectively inhibit epithelial lipid uptake while preserving glucose handling. A structurally diverse FAHFA library was screened in HepG2 cells under basal and insulin stimulated conditions. Inhibition of lipid uptake was summarized as area under curve (AUC) of 100 log2 (fold change), where AUC>0 indicates no inhibition versus DMSO controls, and increasingly negative AUC reflects stronger and broader inhibition across different concentrations (Figure 16). Using an arbitrary cutoff of AUC < -25, 7 FAHFAs (10-HAHSA, 10-PAHSA, 12-HAHSA, 12-PAHSA, 12-SAHSA, 12-TAHSA and 12-PAHIA) were shortlisted which demonstrated robust suppressive effect on lipid uptake under both basal and insulin-stimulated conditions (Figure 16A-B). Dose-response profiling (1.56 to 200 pg / mL) of these FAHFAs confirmed their lipid uptake inhibition across broad ranges and is largely insulinindependent. Many FAHFAs also showed non-monotonic (U-shaped) responses at higher doses, likely due to solubility limits at higher doses given their hydrophobic nature. Among these, 12-SAHSA and 12-TAHSA showed minimal cytotoxicity and negligible impact on glucose uptake, nominating them as parent scaffolds for lead optimization(Figure 16C). The screen suggested that subtle structural features, especially branch position, drive potency, motivating a focused structure-activity analysis.

[0524] To guide rational optimization and explore structure-activity relationship of FAHFAs, it was tested whether branched position and backbone geometry predict FAHFA potency against lipid uptake. Pairwise Spearman correlation between structural properties such as branching carbon position, hydroxy fatty acid (HFA) backbone length, fatty acyl (FA) branch length, purity, yield and the AUC of lipid uptake assays was performed. While no single descriptor fully explained activity, a clear structure-activity relationship emerged. CIO- and C12- branched FAHFA regioisomers consistently outperformed other positions, implying that their 3D presentation favors interaction with lipid-processing molecular targets (Figure 16B, 2D). Insulin sensitivity varied with branching position. For instance, C2-branched regioisomers were more potent at basal conditions while C3-branched FAHFAs exhibited stronger inhibitory effect with insulin, whereas the C10 / C12 effects were largely insulin-insensitive. To translate this structure activity relationship into drug-like, orally stable candidates at scope, a modular chemistry platform was built enabling rapid access to regio-defined and bioisosterica lly stabilized FAHFAs.

[0525] Scalable synthetic platform yields potent FAHFA analogues

[0526] To unlock chemical space and install oral-stability features without sacrificing selectivity, a scalable, modular synthesis was established that accelerates design-make-test. The modular strategy involves a streamlined four-step sequence, delivered 49 FAHFAs spanning chain lengths and regiochemistries with high regioselectivity and purity (Figure 2A-B). It begins with a stereoselective Grignard addition to fatty aldehydes, yielding diverse hydroxy-fatty acids in high yield (Figure 2A, condition a). These intermediates are then converted to methyl esters via reflux in methanol with thionyl chloride, yielding methyl hydroxy-fatty acids (Figure 2A, condition b). Subsequent esterification of the secondary alcohol with desired acyl chloride (in pyridine) affords FAHFA methyl esters (Figure 2A, condition c), and a final selective hydrolysis of the methyl ester yields the target FAHFA (free acid) compound in good overall yield (Figure 2A, condition d).

[0527] Building on the 12-SAHSA / 12-TAHSA scaffolds, bioisosteric linkages (amide, 1,2,3-triazole) was introduced to overcome the acid- / enzyme-labile ester while retaining lipidselective activity. A panel of 15 derivatives was synthesized (Figure 2B), spanning carbonate ester (e.g. 12- DCAHSA and 12-HCAHSA), sulfonate ester (e.g. 12-DSAASA and 12-HSAHSA), amide (e.g. 12-TAASA and 12-SAASA) and 1,2,3-triazoles (e.g. 12-DDTZSA 143 and 12-HDTZSA) linkages, plus deuterated and enantioenriched variants (e.g. 12-GFTAHSA, 12-cfSAHSA, (7? J-12-TAHSA and (7?)-12-SAHSA) to probe for their stability and activity.

[0528]

[0529] Across HepG2 and AML12 cells, 12-SAASA (an amide analog of 12-SAHSA), 12-TAASA (an amide analog of 12-TAHSA), and 12-HDTZSA (a triazole analog of 12-HSAHSA) emerged as standout leads. These derivatives robustly inhibited lipid uptake, spared glucose uptake, and showed low cytotoxicity (Figure 17A-B). In vitro acid-challenge assays (pH 1-2) confirmed improved gastric stability for bioisosteres; 12-TAASA and 12-HDTZSA were ~12.6% and 9.2% more resistant to hydrolysis than 12-SAHSA, respectively (Figure 17B). Time course acid-stability assays at pH 1.5 showed that 12- TAASA, the most acid-resistant derivative, retained 98-95% of parent compound over 48 h, whereas the parent ester 12-TAHSA declined to ~85% by 48 h, confirming superior gastric stability of the amide analogue. Several variants with narrow activity windows, off-target glucose effects, or dose limited toxicity were deprioritised. Collectively, these data identified 12-TAASA, 12-157 SAASA, and 12-HDTZSA as intestine-anchored candidates, which were advanced to in-vivo testing to determine if oral FAHFAs can deliver semaglutide-like metabolic and hepatic benefits in LIDPAD mice.

[0530] Oral FAHFAs deliver semaglutide-like metabolic and hepatic benefits in LIDPAD mice With these optimized, gut-anchored analogues in hand, it was investigated whether oral dosing in LIDPAD mice can deliver semaglutide-like improvements in weight, glycemia, and hepatic steatosis. Stabilized FAHFAs can modulate gut-to-liver lipid flux via an intestine anchored checkpoint using C57BL / 6J mice on the Liver Disease Progression Aggravation Diet (LIDPAD), a high-fat, high-calorie model that accelerates MASLD. After diet induction, animals were randomized to once daily by oral FAHFAs, including parent FAHFAs (12-SAHSA, 12-TAHSA) and synthetic derivatives (12-SAASA, 12-168 TAASA, 12-HDTZSA; 100 mg / kg) for 4 weeks. Efficacy was benchmarked against semaglutide (100 pg / kg subcutaneous, twice weekly) and untreated LIDPAD controls.

[0531] Under these conditions, 12-TAASA and 12-HDTZSA consistently blunted weight gain, despite continued LIDPAD diet, to a degree comparable to semaglutide (Figure 11A) and improved glucose tolerance and insulin sensitivity over the treatment window (Figure 11B), indicating that an oral, gut-first FAHFA regimen can approach the metabolic control achieved by a potent injectable incretin mimetic.These metabolic gains translated to liver-centric endpoints. Mice receiving 12-TAASA, 12-HDTZSA, or 12-SAHSA showed lower liver mass and visceral white adipose tissue (WAT) mass than untreated LIDPAD mice, consistent with reduced hepatic lipid load and diminished visceral adiposity. Histological analysis confirmed that FAHFA treatment protected the liver from severe lipid accumulation observed in untreated LIDPAD mice (Figure 11C-D). Although the parent FAHFAs (12-SAHSA, 12-TAHSA) were modestly less effective on body-weight trajectory, they still reduced visceral adiposity and improved glucose tolerance (Figure 11B-D), supporting a class-wide, gut-first mechanism with chemistry-enabled gains in potency and robustness.

[0532] Safety and tolerability were favorable. There were no adverse changes in heart, spleen, or kidney weights and no treatment-related histopathology in these organs across groups. Collectively, these data show that orally dosed, stabilized FAHFAs deliver semaglutide like control of weight trajectory and glucose homeostasis while ameliorating hepatic steatosis and do so under thermoneutral conditions that minimize cold-stress confounds at the gut-liver axis.

[0533] Gut microbiome remodeling and SCFA enrichment by synthetic FAHFAs

[0534] To map proximal mechanisms underlying the metabolic benefits, in vivo fate and distribution of orally dosed FAHFAs was characterized. After a single oral dose (100 mg / kg), both parent and synthetic FAHFAs were detectable in feces but not in plasma, consistent with minimal systemic absorption (Figure 13A). Consistent with fecal retention and negligible systemic exposure, HPLC analysis failed to detect compound-matched peaks in plasma for all tested FAHFAs under our conditions. Notably, fecal levels of 12-SAASA, 12-TAASA and 12-HDTZSA peaked at ~24 h and returned to baseline by 48 h, whereas parent 12-SAHSA and 12-TAHSA remained elevated at 48 h (Figure 13A), suggesting greater intraluminal processing or uptake for the engineered derivatives.

[0535] Interestingly, 16S rRNA gene sequencing of the gut microbiota revealed distinct beta diversity clusters for 12-TAASA or 12-HDTZSA-treated mice, clearly separated from LIDPAD control. Parent FAHFAs overlapped with controls (Figure 13B). At the genus level, 12-TAASA and 12-HDTZSA consistently depleted Romboutsia and Olsenella (genera associated with inflammation in high-fat diet context) while enriching Blautia, a canonical short-chain fatty acid (SCFA)-producing bacteria. 12-SAASA produced a similar increase in Blautia and decrease in Olsenella, but was less effective against Romboutsia. These compositional changes suggest that synthetic FAHFAs create a gutenvironment favoring beneficial microbes over potentially deleterious ones. Over the same window, comparable shifts were not observed with semaglutide, aligning with its predominantly host-mediated mechanism in our system.

[0536] Metagenomic (PICRUSt2-based) inferences pointed to coordinated upregulation of pathways converting fiber-derived substrates and glycolytic intermediates (pyruvate, lactate) into acetate, propionate, and butyrate under 12-TAASA and 12-HDTZSA (Figure 13D-E). Taxonomically, this functional gain paralleled selective expansion of Blautia, Lachnospiraceae NK4A136, and Roseburia. These taxa and their SCFAs are linked to improve energy balance, gut barrier integrity, and anti-inflammatory tone, providing a possible conduit from microbiome remodeling benefits observed.

[0537] Together, these data indicate that chemically stabilized FAHFAs remodel the gut ecosystem toward SCFA-producing consortia while suppressing pro-inflammatory genera, changes that are coherent with their fecal-retentive PK and distinct from semaglutide's host-centric route. It is therefore posit that microbiome-mediated SCFA enrichment is a major contributor to FAHFA efficacy, and an integral component of the gut-to-liver lipid-flux checkpoint engaged by these agents.

[0538] 12-TAASA directly suppresses intestinal lipid absorption programs

[0539] Having shown that synthetic FAHFAs remodel the microbiome toward SCFA production, it is investigated whether 12-TAASA, the top-performing compound, also acts directly on the intestinal epithelium to curb lipid entry. Transcriptomic profiling of small intestinal mucosa from treated mice revealed a distinct gene expression signature induced by 12-TAASA relative to other FAHFAs (Figure 14A). While FAHFAs broadly stimulate genes involved in intestinal cell proliferation, potentially aiding epithelial turnover and gut barrier maintenance, suggesting a general trophic effect on the mucosa. 12-TAASA (and to a lesser extent its parent 12-SAHSA) uniquely downregulated genes related to fibrogenesis and lipid metabolism (Figure 14B). Pathway analysis showed 12-SAHSA mainly suppressed adipokine signaling, whereas 12-TAASA broadly repressed the expression of genes involved in the transport and processing of dietary lipids (cholesterol, fatty acids, ketone bodies, steroids) (Figure 14C). Other FAHFAs displayed orthogonal effects, e.g., 12-TAHSA and 12-SAASA upregulated amino acid metabolism genes, and 12-HDTZSA modulated immune response genes, but these were not directly related to lipid absorption. Together, these data point to a potent local inhibition of epithelial lipid-uptake programs under 12-TAASA, consistent with its ability to prevent diet-induced fat accumulation.Using a protein-protein interaction network analysis of the genes most strongly downregulated by 12-TAASA, several hub genes were identified that could serve as central regulators of intestinal lipid absorption. Top hub candidates included Cd44, Aldh1a1, Apoa1, Ugt1a1, Cyp2d26, Hspa1b, Hmgcs2, Fabp2, Dgat2 and Tlr2 (Figure 14D). Five of these (Cd44, Apoa1, Fabp2, Dgat2, Tlr2) encode cell-surface or membrane-associated proteins, suggesting that 12-TAASA could act at the membrane interface to blunt lipid import and trafficking. To test if 12-TAASA physically interacts with intestinal receptors or transporters, a biotinylated 12-TAASA probe was synthesised and ligand pull-down assays were performed on intestinal mucosal lysates (Figure 14E). Proteomic analysis of pulled-down proteins identified 468 putative binding partners, including 134 membrane proteins enriched for lipid metabolism / transport / endocytosis (Figure 14F). Crucially, Cd44, prioritized by transcriptomic hub analysis, was confirmed among the pulled-down targets (Figure 14G). Two additional overlaps (Chp1 and Hspa1 family proteins) linked the binding set to downregulated transcripts, and membrane interactors formed a dense network with 12-TAASA-suppressed genes, indicating convergence of binding and gene-program remodeling.

[0540] To examine whether 12-TAASA directly engages human CD44, it was docked to the hyaluronic acid binding domain (CD44-HABD). Two independent algorithms, Attracting Cavities 2.0 and AutoDock Vina, converged on potential sites in which 12-TAASA occupies a pocket adjacent to the hyaluronan groove, stabilized by hydrogen bonds and hydrophobic contacts (Figure 18A). This pocket overlaps the tetrahydroisoquinoline (THIQ) pharmacophore site implicated in CD44 inhibition. In the Attracting Cavities 2.0 model, the fatty-acid tail of 12-TAASA extended into the HA-binding groove, further supporting direct engagement.

[0541] Functionally, CD44 knockdown by siRNA in the human colon epithelial cell line NCM460 markedly reduced lipid uptake, similar in magnitude to 12-TAASA treatment (Figure 18B). Importantly, co-treatment with 12-TAASA and si-CD44 did not yield additional inhibition, consistent with 12-TAASA acting through CD44 blockade. Given the roles of CD44 in fatty-acid translocation and epithelial inflammatory signaling, its engagement by 12-TAASA offers a mechanistic basis for limiting dietary fat uptake while tempering local inflammatory tone. The intestinal transcriptome plus direct target engagement supports a model in which 12-TAASA applies a membrane-level "brake" (CD44-centered, with allied transporters like FABP2 / DGAT2) on intestinal lipid uptake. Togetherwith its microbiome-driven SCFA enrichment, 12-TAASA therefore exerts dual gut-anchored control over gut-to-liver lipid flux, consistent with its robust metabolic benefits in vivo.

[0542] Discussion

[0543] MASLD progression is tightly coupled to the rate of lipid delivery from intestine to liver. This work identifies stabilized FAHFAs as oral agents that act locally at the gut node to lower this flux. A modular synthesis platform broadened FAHFA chemical space and enabled bioisosteric stabilization of the ester, yielding gut-retentive analogues that preserve lipid-selective biology. From this platform, 12-TAASA and 12-HDTZSA emerged as optimized leads. On oral dosing in LIDPAD mice, a diet-induced mouse model that mimics key features of human MASLD, these analogues reduced weight gain and hepatic / visceral lipid burden and improved glycemic control, outcomes most consistent with reduced gut-to-liver lipid import rather than downstream oxidation alone.

[0544] Mechanistically, FAHFAs deploy two intestinal "brakes" on flux. First, they remodel the microbiome toward SCFA-producing consortia, cohering with fecal retention and a lumen focused exposure profile. The combination of robust acid stability and fecal detection without plasma peaks further supports a gut-anchored mechanism for the engineered FAHFAs. Second, 12-TAASA engages epithelial programs that down-tune lipid-handling pathways and binds a membrane target set centered on CD44, with allied transport / trafficking proteins (e.g., FABP2, DGAT2). Together, the microbiome to SCFA axis and the epithelial CD44-centered brake are consistent with lower chylomicron formation / entry and thus less hepatic lipid influx, aligning proximate mechanisms with organismal benefits in MASLD.

[0545] This flux-aware view places FAHFAs alongside, yet mechanistically distinct from, host directed strategies. Rather than relying on systemic endocrine or hepatic reprogramming, FAHFAs act upstream at the gut interface where flux is set. The therapeutic implication for MASLD is practical. FAHFAs can be positioned to (i) blunt lipid input during periods of higher dietary fat exposure, (ii) pair with host-acting agents that modulate bile-acid control or increase peripheral oxidation, and (iii) provide biomarker-anchored pharmacodynamics in the gut (stool FAHFAs and SCFAs; epithelial target signatures).

[0546] This work also clarifies how FAHFA derivatives couple host and microbiome pathways to deliver metabolic benefits. The pronounced gut microbiota remodeling observed with12-TAASA and 12-HDTZSA addresses a gap in the literature, where FAHFA-microbiome interactions were incompletely defined. It was consistently found enrichment of SCFA-producing genera (e.g., Blautia, Roseburia) and depletion of pro-inflammatory taxa (e.g., Romboutsia).

[0547] SCFAs such as acetate and butyrate improve insulin sensitivity, induce satiety, and strengthen gut barrier. Thus, FAHFA-induced microbiome changes likely contribute materially to anti obesogenic effects, aligning with concepts that treat metabolic disease via microbiome modulation. Native FAHFAs may also signal through gut microbes; indeed, prior studies suggested that 5- and 9-PAHSA can activate GPR120 on intestinal cells to promote antimicrobial peptide release, thereby indirectly shaping microbial communities. By introducing amide bonds and other modifications, 12-TAASA appears to amplify these microbiome-mediated effects, by prolonging FAHFA presence in the distal gut where most microbes reside. Enhanced SCFA production in 12-TAASA-treated mice provide a plausible link between the microbiota shifts and the observed improvements in adiposity and glycemic control.

[0548] Concurrently, 12-TAASA directly targets the host's intestinal nutrient handling. Transcriptomic and proteomic analyses converged on CD44 as a key mediator of 12-TAASA action in the gut. CD44 has been implicated in fatty acid uptake and cholesterol metabolism in enterocytes. By binding CD44 (and possibly other surface proteins like ApoA1 and FABP2), 12-TAASA may interfere with dietary fat absorption via a receptor-mediated route rather than broad enzymatic blockage. This mode is reminiscent in outcome, but distinct in mechanism, from intestinal lipase inhibitors (e.g., orlistat) or bile-acid sequestrants, delivering a more targeted "brake" on lipid uptake, consistent with reduced expression of lipid-transport genes and lower fat deposition in treated animals. Notably, this dual mechanism-of-action, microbiome and host receptors, is unusual among metabolic drugs. Current medications typically act either on the host endocrine axes (e.g., GLP-1 analogues on appetite and insulin) or on nutrient absorption (e.g., pancreatic lipase inhibition) without intentional microbiome modulation. FAHFAs, by engaging both domains, may inaugurate a two-pronged therapeutic class.

[0549] Beyond weight loss and lipid metabolism, our findings hint at broader health benefits. FAHFAs appear to stimulate intestinal epithelial proliferation, potentially supporting mucosal healing and barrier integrity, useful in condition such as inflammatory bowel disease or increased intestinal permeability. Others have reported that FAHFAs enhance insulin-stimulated glucose uptake in muscle and fat tissues, suggesting potential asinsulin sensitizers or adjuncts in diabetes management. As lipids, FAHFAs (especially stabilized analogues) are membrane-permeable and may distribute to multiple tissues, potentially exerting systemic anti inflammatory effects. Taken together, these properties position FAHFAs as versatile therapeutic candidates for metabolic syndrome, obesity, dyslipidemia, insulin resistance, and gut health.

[0550] Despite this promise, several considerations remain. Chronic inhibition of dietary lipid absorption, if excessive, could lead to fat-soluble vitamin (A, D, E, K) and essential fatty acid deficits. Long-term nutritional monitoring will be needed in therapeutic contexts. The preclinical safety profile of synthetic FAHFAs is favorable, but formal toxicological evaluations are required to establish dosing margins and exclude rare toxicities, if any. Inter-individual microbiome variability may shape FAHFA efficacy. Personalized microbiome analysis could identify patients most likely to benefit or reveal probiotic cotherapies that enhance efficacy. Finally, while our chemistry simplifies FAHFA production, manufacturing scale-up and formulation stability will be essential for translation.

[0551] In summary, this platform unlocks the therapeutic potential of FAHFAs by overcoming longstanding synthetic and stability barriers and enabling systematic exploration of structure activity relationships. Strategic bioisosteric bond modifications enhance metabolic efficacy and acid stability of selected regioisomers, critical for oral, gut-localized action. The discovery of 12-TAASA exemplifies this advance, demonstrating dual-action benefits, host epithelial modulation and microbiome remodeling, with efficacy that, under our conditions, matches the magnitude of benchmark comparators in diet-induced obesity models. Beyond metabolic diseases, this platform redefines FAHFA biology, providing a framework to dissect how lipid structure governs functions, while advancing flux-aware intervention for MASLD.

[0552] Procedure A. General procedure for preparation of the n-hvdroxy fatty acids, (step a) A stirred solution of compound 1 (2.0 mmol, 1.0 equiv.) in dry THF (20 mL) underwent the addition of the selected Grignard reagent (6.0 mmol, 3 equiv.) under a nitrogen atmosphere at 0 °C. Subsequently, the reaction mixture was stirred for 8 hours at room temperature, and saturated aqueous NH4CI solution (20 mL) was added. The resulting mixture was then extracted with DCM (20 mL x 3). The combined organic extracts were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude compound was purified through silica gel columnchromatography using a DCM: Methanol solvent system to yield the N-hydroxy fatty acids.

[0553] General procedure for preparation of the methyl esters of the n-hvdroxy fattv acids, (step bl To a solution of N-hydroxy fatty acids 2 (1.5 mmol, 1 equiv.) in MeOH (10 mL), SOCI2 (1.95 mmol, 1.3 equiv.) was slowly added. The reaction mixture was heated at reflux for 5 hours and then evaporated to remove MeOH. The residue was diluted with ethyl acetate (30 mL) and quenched with water (20 mL). A saturated aqueous solution of NaOH was added to adjust the pH to 8-9. The organic layer was separated, washed with water and brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The crude compound was purified via silica gel column chromatography using Hexane: EtOAc to yield the methyl ester of the N-hydroxy fatty acids.

[0554] General procedure for the preparation of the methyl ester of FAHFAs. (step c) To a stirred solution of the N-hydroxy methyl ester compound 3 (1.0 mmol, 1.0 equiv.) in dry dichloromethane (10 mL), dry pyridine (5.0 mmol, 5.0 equiv.) was added at 0 °C, and the solution was stirred for 15 minutes. Acyl chloride (1.8 mmol, 1.8 equiv.) was added to the reaction mixture. The reaction was allowed to warm to 23 °C and stirred for 12 hours. The unreacted acyl chloride and pyridine were quenched with HCI (1.0 N, 10 mL). The mixture was extracted with dichloromethane (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane: EtOAc to give the products.

[0555] General procedure for preparation of the FAHFA families, (step d) The methyl ester of FAHFAs 4 (0.5 mmol, 1 equiv.) was dissolved in THF: H2O (8 mL: 8 mL, 1:1) and cooled to 0 °C for 15 minutes before adding solid LiOH (3.25 mmol, 6.5 equiv.) in one portion. The reaction was allowed to warm to 23 °C and stirred for 24 hours. Excess hydroxide was neutralized with 1 N HCI. The resulting product was extracted with DCM (10 mL x 3), and the combined organic extracts were washed with brine (5 mL), dried over sodium sulfate, filtered, and concentrated. The crude products were subsequently purified by silica gel column chromatography, eluting with DCM: Methanol to yield FAHFAs.

[0556] Procedure B. General procedure for preparation of ether derivativesCH <v I w iu 1. KjCOg (1.3 equiv.), OMR SO ®C „ | ' ' v$v,rGOsM€+Alkyl bromide - «-rv?e>^-''5.„.<co:^ ' ‘S ’0 2 LiOH (6.5 eqwv4- THF: H2O (1:1). rt ’ ’a «

[0557]

[0558] i - 12, 15 To a solution of 12-Hydroxy methyl ester compound (1.0 equiv.) in dry DMF (2 M), alkyl bromide (1.2 equiv.) and K2CO3 (1.3 equiv.) were added at r.t. and then the reaction was stirred at 80 °C for 12 hours. The mixture was quenched with NH4CI solution, and extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the methyl esters of ether compounds.

[0559] Methyl esters of ether compounds (1.0 equiv.) were dissolved in THF: H2O (1: 1) at 0 °C for 15 minutes before solid lithium hydroxide (LiOH, 6.5 equiv.) was added. The reaction was stirred at room temperature for 24 hours, then neutralized with 1 N HCI. The product was extracted with DCM (3 x 10 mL), washed with brine (5 mL), dried (Na2SO4), filtered, and concentrated. Purification was via silica gel chromatography (DCM / methanol) to yield products.

[0560] Procedure C. General procedure for preparation of deuterated analogues

[0561] a V 1. NaBD^ (2.0 equiv / ). MeOH, RTxDJ ', Me. A.rCO2Me. --. -. A^C°3H

[0562]

[0563] •'<5p 2. Procedure A: Step c and step d 5 10 i = 11, 15 To a solution of methyl 12-oxooctadecanoate (1.0 equiv.) in dry MeOH (IM), NaBD4(2.0 equiv.) was added at rt. And then the reaction was stirred at RT for 1.5 hours. The mixture was quenched with NH4CI solution, which was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the methyl 12-d-hydroxyoctadecanoate. The deuterated derivative of FAHFA was then synthesized according to Step c and Step d.

[0564] Procedure D. General procedure for preparation of (R)-FAHFAs. The 12-HSA (0.2 mmol), vinyl myristate (0.3 mmol) or vinyl stearate (0.3 mmol), Novozym 435 (10 mg) and diisopropyl ether (DIPE, 1 ml) were added into a 4 ml reaction tube. The reaction mixture was stirred (400 rpm) at 40 °C for 24 h. The solvent was removed, and the residue was purified by flash chromatography on silica gel to afford the desired product.Procedure E: General procedure for preparation of amide derivatives

[0565] 1. NaBHsCN (1,2 equiv.), O NH4AG (10 equiv), MeOH. RT Hhf HpMe Me.. XrCO, H 2. Alkanoyl chloride (1.8 equiv)

[0566]

[0567] ' '5 10b bEt3N (3.0 equiv ), DOM, RT i = 11. 15

[0568] 3. Procedure A; step d

[0569] To a solution of methyl 12-oxooctadecanoate (1.0 equiv.) in dry MeOH (0.5 M), NaBHsCN (1.2 equiv.) and NH4AC (10 equiv.) were added at rt. And then the reaction was stirred at RT for 12 hours. The mixture was quenched with NH4CI solution, which was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using DCM / MeOH to give the methyl 12-aminooctadecanoate.

[0570] To a solution of methyl 12-aminooctadecanoate (1.0 equiv.) in dry DCM (10 M), dry EtsN (3.0 equiv.) was added at 0 °C. After 15 minutes of stirring, acyl chloride (1.8 equiv.) was introduced, and the reaction was stirred at room temperature for 12 hours. The unreacted acyl chloride and EtsN were quenched with HCI (1.0 N, 10 mL). The mixture was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the products. The amide derivative of FAHFA was then synthesized according to Procedure A: Step d.

[0571] Procedure F: General procedure for preparation of sulfonate ester analogues

[0572] O

[0573] 1. Sulfonyl chlo de (1.2 equiv) OH EtsN (1.5 equiv ). DCM, RT Mib,. CO2Me - Me. XrCO, H 2. Procedure A; step d

[0574]

[0575] 10

[0576]

[0577] qo i - 11, 15 To a solution of methyl 12-hydroxy methyl ester compounds (1.0 equiv.) in dry DCM (1 M), dry EbN (1.5 equiv.) was added at 0 °C. After 15 minutes of stirring, sulfonyl chloride (1.2 equiv.) was introduced, and the reaction was stirred at room temperature for 12 hours. The unreacted sulfonyl chloride and EtaN were quenched with HCI (1.0 N, 10 mL). The mixture was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the products. The sulfonate ester of FAHFA was then synthesized according to Procedure A: Step d.Procedure G: General procedure for preparation of carbonate ester analogues

[0578] O

[0579] 1. Chlorofomiate('1.8 equiv)OHPyridine (5.0 equiv ), DCM, RT CT Me. M Ar „CO2Me ’7 z7,7 P7r.oced.ure A 7.: s 7te.p ’ d7. *Me. < > A■.K,r-1C0Q2H

[0580]

[0581]

[0582] l - 10, 14 To a solution of methyl 12-hydroxy methyl ester compounds (1.0 equiv.) in dry DCM (1 M), dry pyridine (5 equiv.) was added at 0 °C. After 15 minutes of stirring, alkyl chloroformate (1.8 equiv.) was introduced, and the reaction was stirred at room temperature for 12 hours. The unreacted chloroformate and pyridine were quenched with HCI (1.0 N, 10 mL). The mixture was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the products. The carbonate ester of FAHFA was then synthesized according to Procedure A: Step d.

[0583] Procedure H. General procedure for preparation of 1,2,3-triazoles.

[0584] tc«q....9^.’..??..”?.. ►, XO5Ms. ----...-■X,x, SO.iMe i Aftyne I CuCOAck i DIPSA

[0585] x CO-H ' ’"’s " w

[0586]

[0587] i:: W„ ’« > «■ To a round-bottom flask equipped with a magnetic stir bar was charged with methyl esters of the 12-hydroxy fatty acids (1 mmol, 1 equiv), CBr4 (1.5 mmol, 1.5 equiv), PPhs (1.5 mmol, 1.5 equiv), and DCM (10 ml). The reaction mixture was stirred at 30 °C for 24 h. The solvent was removed and the residue was purified by flash chromatography on silica gel to afford the methyl 12-bromooctadecanoate.

[0588] A 10 ml round-bottom flask (RBF) was charged with a solution of methyl 12-bromooctadecanoate (0.8 mmol, 1 equiv) and sodium azide (4 mmol, 4 equiv) in DMF (4 ml). The reaction mixture was stirred at 100 °C for 12 h, and then quenched with water, extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and evaporated to dryness. The crude product was purified by flash chromatography on silica gel to give the azide intermediate.To a solution of azide (0.4 mmol, 1 equiv.) in acetonitrile (1.5 ml), alkyne (0.4 mmol, 1 equiv.) and Cu(OAc)2 (0.02 mmol, 0.05 equiv.) were added and stirred until the Cu(OAc)2completely dissolved. N, N-diisopropylethylamine (DIPEA, 0.04 mmol, 0.1 equiv.) was then added, and the reaction mixture was allowed to stir at 45 °C for 1 hour. The solvent was then removed under reduced pressure, and the crude product was purified by flash chromatography.

[0589] The 1,2,3-triazoles (0.25 mmol, 1 equiv.) were dissolved in THF:H2O (4mL: 4 mL, 1: 1) and cooled to 0 °C for 15 minutes before adding solid LiOH (1.63 mmol, 6.5 equiv.) in one portion. The reaction was allowed to warm to 23 °C and stirred for 24 hours. Excess hydroxide was neutralized with 1 N HCI. The resulting product was extracted with DCM (10 mL x 3), and the combined organic extracts were washed with brine (5 mL), dried over sodium sulfate, filtered, and concentrated. The crude products were subsequently purified by silica gel column chromatography, eluting with DCM: Methanol to yield products.

[0590] Cell culture. Murine healthy hepatocytes (AML12) and human hepatocellular carcinoma cells (HepG2) were obtained from ATCC. AML12 cells were cultured in DMEM / F12 supplemented with 10% fetal bovine serum (FBS), lx insulin-transferrin-selenium (ITS; Gibco, MA, USA), and 40 ng / mL dexamethasone. HepG2 cells were cultured in DMEM supplemented with 10% FBS. All cells were maintained in a humidified 37°C incubator with 5% CO2.

[0591] Lipid absorption assay. To examine the effect of FAHFAs on lipid absorption, cells were resuspended in serum-free DMEM and seeded into each well of a 96-well plate, followed by overnight incubation. The next day, the cells were treated with varying concentrations of FAHFAs with or without insulin (100 nM) for 15 minutes. Control wells were treated with DMSO or disulfiram (known for its anti-obesity effect). Blank wells contained culture medium without any cells. After 15 minutes, 100 pL of BODIPY 500 / 510 Cl, C12 (MedChemExpress, NJ, USA), a fluorescent fatty acid analog for lipid trafficking assay, was added to a final concentration of 1 pM. The 96-well plate was then transferred to a fluorescence microplate reader for kinetic measurement of the fluorescent signals with a maximum excitation wavelength of 485 nm and emission wavelength of 515 nm every minute for 20 minutes. For data analysis, the signal from blank wells was averaged and subtracted from those treated with DMSO, disulfiram, or FAHFAs. The rate of change of the fluorescent signal within the linear range, indicating the rate of lipid uptake, was calculated.Glucose uptake assay. For glucose uptake assay, cells were seeded at the same density as lipid adsorption assay in glucose- and serum-free overnight. The next day, the cells were treated with varying concentrations of FAHFAs with or without insulin (100 nM) for 15 minutes the next day while control wells were treated with DMSO or disulfiram. Blank wells contained culture medium without any cells. After 15 minutes, 2-NBDG (100 pM) was added to each well and incubated for 30 minutes to enable its uptake. After that, the culture supernatant was aspirated. Each well was washed with PBS twice and resuspending in 100 pL of PBS. 2-NBDG taken up by the cells was measured with a fluorescence microplate reader set to a maximum excitation wavelength of 485 nm and emission wavelength of 515 nm. For data analysis, the signals from blank wells were averaged and subtracted from those treated with DMSO, disulfiram, or FAHFAs. To assess glucose uptake, the differences between fluorescent signals from different treatments were computed.

[0592] Acid stability assay. FAHFAs (0.2 mg) were subjected to acid hydrolysis in 3 mL of HCI solution (pH 1.5) under continuous stirring for 12 hours at room temperature. The mixture was then extracted with DCM (5 mL). The organic phase was carefully separated and subsequently concentrated to dryness using a rotary evaporator. The resulting residue was reconstituted in 1 mL of isopropyl alcohol to prepare a homogeneous solution for quantitative analysis by HPLC.

[0593] Viability assay. Cells were seeded in a 96-well plate at a density of 5000 cells / well and incubated overnight. The cells were then treated with increasing concentrations of FAHFAs (1 to 200 pg / mL) for 48 hours. Cell viability was determined using alamarBlue Cell Viability Reagent (Thermo Scientific, MA, USA) following the manufacturer's instructions.

[0594] In vivo mouse experiment. Male C57BL / 6J mice aged between 8-10 weeks were given free access to LIDPAD for four weeks to induce obesity and metabolic syndrome (PMID: 38952069). Control group was given a matched control diet. The mice were housed at a thermoneutral condition (30±2 °C) and a 12-hour darklight cycle throughout the experiment. For the treatment groups, mice were treated with either semaglutide subcutaneously (100 pg / kg; twice per week), 12-SAHSA, 12-TAHSA, 12-SAASA, 12-TAASA or 12-HDTZSA orally (100 mg / kg; daily) while feeding on LIDPAD diet. After the 4-week treatment, an intraperitoneal glucose tolerance test was performed by injecting 2.0 g / kg of glucose intraperitoneally after 16 h of fasting. Blood glucose concentrationswere measured up to 120 min after injection from the tail vein using a glucometer (Accucheck Performa, Roche, USA). Animals were euthanized using xylazine / ketamine overdose prior to tissue collection. Collected tissues were fixed with 4% PFA or snap frozen with liquid nitrogen for subsequent analyses. For pharmacokinetic studies, mice received a single oral dose of FAHFA (3 mg), with stool and blood samples collected at 24- and 48-hours post-gavage for FAHFA quantification. All experiments were carried out following the guidelines of the Institutional Animal Care and Use Committee (IACUC) (A21073).

[0595] FAHFA extraction from mouse stool and blood plasma and quantification. Mouse stool samples were stirred using DCM for 12 hours at ambient temperature. The suspension was subsequently filtered to remove particulate matter, and the solvent was concentrated to dryness via rotary evaporation. The dried residue was reconstituted in 1 mL of isopropyl alcohol to yield a homogenized solution, which was then subjected to quantitative analysis by HPLC.

[0596] Blood plasma was stirred using DCM for 12 hours at ambient temperature. The reaction mixture was then extracted with DCM (5 mL). The organic phase was carefully separated and subsequently concentrated to dryness using a rotary evaporator. The resulting residue was reconstituted in 1 mL of isopropyl alcohol to prepare a homogeneous solution for quantitative analysis by HPLC.

[0597] Histopathological analysis. Liver and white adipose tissues (WAT) were fixed in 4% neutral buffered formalin and embedded in paraffin. Sections (5 pm) were stained with hematoxylin & eosin (H& E) and picrosirius red (PSR) to evaluate tissue histology and fibrosis, respectively. Microscopic images were captured using an Axioscan Z1 (Zeiss, Germany) at 20x magnification. Liver pathology was evaluated using the steatosis, activity, and fibrosis (SAF) scoring system. Adipocyte size and lipid accumulation in WAT sections were quantified using Image!.

[0598] Gut microbiome 16S rRNA sequencing and analysis. Total bacterial DNA was extracted from mouse fecal samples using the ZymoBIOMICS DNA Miniprep Kit (Zymo Research, CA, USA). The V3-V4 region of 16S rRNA was amplified and sequenced on a DNBSEQ-T7 platform (BGI, China), generating 250 bp paired-end reads. Raw reads were processed using DADA2, with amplicon sequence variants (ASVs) mapped to the SILVA database for taxonomic annotations. Microbial diversity was computed using phyloseq. Differentially abundant genera were identified via ALDEx2. ALDEx2 (Iog2 fold change > ±1 and effect size > ±1). Metagenomic functional predictions were generated withPICRUSt2. Transcriptomic analysis of intestinal mucosa. Intestinal mucosae were harvested by scraping the luminal surface of the small intestine. Total RNA was extracted using the Pure-NA Fastspin Total RNA Extraction Kit (Everlife, Singapore).

[0599] RNA sequencing libraries were prepared and sequenced on a DNBSEQ-T7 platform (BGI, China) with 150 bp paired-end reads. Reads were aligned to the Mus musculus reference genome (GRCm39) using HISAT2. Gene counting was performed using FeatureCounts while differential expression analysis was done using DESeq2.

[0600] Gene quantification was performed with FeatureCounts, and differential expression analysis was carried out using DESeq2. Genes with log2 fold-change > ±1 and adjusted p-value <0.05 were designated as differentially expressed genes (DEGs). Hub gene analysis was conducted via Cytoscape using cytoHubba to identify critical regulatory genes affected by FAHFAs.

[0601] Biotinylated 12-TAASA pull-down and proteomic analysis. A biotin-conjugated derivative of 12-TAASA was synthesized by amide coupling of biotin to the carboxyl terminus of 12-TAASA through a short linker.

[0602] Intestinal mucosa lysates were prepared using Mammalian Protein Extraction Reagent (Thermo Fisher Scientific, MA, USA) containing Halt™ Protease and Phosphatase Inhibitor Cocktails. The lysates were centrifuged at 10,000 xg for 15 minutes and the supernatant was collected for protein quantification. Lysates (10 pg protein) were incubated overnight at 4°C with biotin-conjugated 12-TAASA (1 pg), facilitating the formation of FAHFA-protein complexes. Complexes were subsequently captured using Pierce™ Streptavidin Magnetic Beads (0.25 mg; Thermo Fisher Scientific), washed thrice with TBST (0.1% Tween 20), and eluted with a low-pH buffer (0.1 M glycine, pH 2). The eluate was neutralized with 1 M Tris (pH 7.5) followed by proteomic profiling using LC-MS / MS analysis.

[0603] The eluate was centrifuged at 12,000 rpm for 15 minutes, and the protein concentration in the supernatant was measured using the BCA assay. A total of 10 pg of lysate was processed using the SISPROT method, a spin-tip-based sample preparation technique. Briefly, the SISPROT device was constructed by packing several C18 disk plugs (3M Empore, USA) into a standard 200 pL pipette tip, followed by the addition of mixed strong cation exchange (SCX) and strong anion exchange (SAX) beads (POROS, Applied Biosystems, USA) in a 1:1 ratio. All subsequent steps— including sample loading, proteinreduction, alkylation, enzymatic digestion, and desalting— were performed on the SISPROT tip. Peptides were then eluted, lyophilized to dryness, and reconstituted in 0.1% (v / v) formic acid for nano-LC-MS / MS analysis using an Orbitrap Exploris 480 mass spectrometer.

[0604] Raw files were searched using Proteome Discoverer (PD) software. The maximum missed cleavage for trypsin digestion was set to 2. The mass tolerance for peptide precursors was 10 ppm and the mass tolerance for fragment ions was 0.02 Da. These readings of every protein from each MS run are derived from the average reading of their isoforms, weighted according to the number of quantifying PSM.

[0605] Molecular

[0606]

[0607] . Ligand receptor interaction between 12-TAASA and human CD44 HABD were examined using Swissdock (52). The ligand (12-TAASA) was imported using a SMILES notation [CCCCCCCCCCCCCC(=O)NC(CCCCCC)CCCCCCCCCCC(=O)O] while the 3D conformation of CD44 (PDB ID: luuh) was obtained from Protein Data Bank in Europe (PDBe). Two docking programs, Attracting Cavities 2.0 and AutoDock Vina were used to anticipate potential binding sites around the hyaluronic acid binding groove. The molecular docking results were imported into Mol* Viewer for 3D visualization.

[0608]

[0609] CD44 knockdown and lipid absorption microscopy Gene silencing of CD44 was achieved by transfecting NCM460 with CD44-targeting duplex siRNA (hs. Ri. CD44.13.1; Integrated DNA Technologies, IA, USA) using Lipofectamine2000 (Thermo Scientific, MA, USA). Control cells were transfected with non-targeting siRNA. Knockdown efficiency was confirmed with immunoblotting based on a published protocol. Antihuman CD44 (H4C4) and anti-human g-tubulin (E7) were obtained from Developmental Studies Hybridoma Bank (IA, USA). After successful CD44 knockdown, the cells were treated with 12-TAASA (100 pg / mL) for 12 h followed by staining with BODIPY 500 / 510 Cl, C12 (1 pM) and Hoechst 33258 (1 pg / mL) for 30 mins. The cells were visualized using Axio Observer 7 (Zeiss, Germany) at 400x magnification. The microscopic images were processed using ImageJ.

[0610] Statistical

[0611]

[0612] . Data are presented as mean ± standard deviation (SD) from at least three independent replicates. One-way ANOVA followed by Tukey's post hoc test was applied for multiple comparisons. Mixed-model ANOVA was used for repeated-measures analyses. Statistical significance was defined as a p-value <0.05.Conclusion. Through in vitro and in vivo screenings, the inhibitory effect of 12-SAHSA, 12-TAASA and 12-HDTZSA on cellular lipid uptake and metabolic dysfunction arisen from chronic high-fat feeding was systematically established. The desirable safety profiles and ameliorative effects on glucose metabolism and adiposity strongly support the potential use of these FAHFAs as food additives or health supplements to achieve weight management and address metabolic syndrome.

[0613] Data characterization of Compounds

[0614] 2-(Hexanoyloxy)octadecanoic acid

[0615] O

[0616] , —J-

[0617]

[0618] This compound was prepared according to the General procedure A.

[0619] 48% yield for 4 steps

[0620] 1H NMR (400 MHz, CDCl3) δ 5.01 (t, J = 6.4 Hz, 1 H), 2.39 (t, J = 7.6 Hz, 2 H), 1.89-1.83 (m, 2 H), 1.70-1.62 (m, 2 H), 1.44-1.26 (m, 32 H), 0.92-0.86 (m, 6 H).

[0621] 13C NMR (100 MHz, CDCl3) δ 176.2, 173.6, 71.8, 34.0, 32.1, 31.3, 31.2, 29.84, 29.80, 29.8, 29.7, 29.5, 29.2, 25.3, 24.6, 22.8, 22.4, 14.3, 14.0.

[0622] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3316.

[0623] 3-(Hexanoyloxy)octadecanoic acid

[0624]

[0625] This compound was prepared according to the General procedure A.

[0626] 55% yield for 4 steps

[0627] 1H NMR (400 MHz, CDCl3) δ 5.22 (ddd, J = 12.8, 7.4, 5.6 Hz, 1 H), 2.63-2.59 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.82 (dd, J = 7.6 Hz, 1 H), 1.68-1.58 (m, 4 H), 1.35-1.25 (m, 30 H), 0.92-0.86 (m, 6 H).

[0628] 13C NMR (100 MHz, CDCl3) δ 180.1, 173.5, 70.2, 34.6, 34.1, 32.1, 31.39, 31.35, 29.9, 29.81, 29.77, 29.68, 29.6, 29.5, 29.48, 25.3, 24.8, 24.5, 22.8, 22.5, 22.4, 14.3, 14.0.

[0629] HRMS (ESI): [M-H] - calcd for C24H45O4397.3318, found 397.3320.

[0630] 5-(Hexanoyloxy)octadecanoic acid

[0631]

[0632] This compound was prepared according to the General procedure A.

[0633] 39% yield for 4 steps

[0634] 1H NMR (400 MHz, CDCl3) δ 4.92-4.86 (m, 1 H), 2.38-2.33 (m, 2 H), 2.29 (t, J = 7.6 Hz, 2 H), 1.67-1.51 (m, 8 H), 1.34-1.25 (m, 26 H), 0.91-0.86 (m, 6 H).

[0635] 13C NMR (100 MHz, CDCl3) δ 179.3, 173.9, 73.5, 34.8, 34.2, 33.9, 33.5, 32.1, 31.5, 31.4, 29.83, 29.79, 29.71, 29.68, 29.6, 29.5, 25.4, 24.9, 24.5, 22.8, 22.5, 20.6, 14.3, 14.1.

[0636] HRMS (ESI): [M-H]⁻ calcd for C24H45O4397.3318, found 397.3328.

[0637] 6-(Hexanoyloxy)octadecanoic acid

[0638]

[0639] This compound was prepared according to the General procedure A.

[0640] 43% yield for 4 steps

[0641] 1H NMR (400 MHz, CDCl3) δ 4.91-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.68-1.50 (m, 8 H), 1.39-1.25 (m, 26 H), 0.91-0.86 (m, 6 H).

[0642] 13C NMR (100 MHz, CDCl3) δ 178.6, 173.9, 73.8, 34.8, 34.3, 33.9, 33.8, 32.1, 31.5, 29.8, 29.72, 29.69, 29.66, 29.5, 25.5, 25.0, 24.7, 22.8, 22.5, 14.3, 14.1.

[0643] HRMS (ESI): [M-H]⁻ calcd for C24H45O4397.3318, found 397.3319.

[0644] 7-(Hexanoyloxy)octadecanoic acid

[0645]

[0646] This compound was prepared according to the General procedure A.

[0647] 56% yield for 4 steps

[0648] 1H NMR (400 MHz, CDCl3) δ 4.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.59 (m, 4 H), 1.54-1.48 (m, 4 H), 1.35-1.25 (m, 26 H), 0.91- 0.86 (m, 6 H).

[0649] 13C NMR (100 MHz, CDCl3) δ 179.1, 173.9, 74.0, 34.8, 34.3, 34.1, 33.9, 32.1, 31.5, 29.78 (2C), 29.71, 29.69, 29.5, 29.1, 25.5, 25.1, 25.0, 24.7, 22.8, 22.5, 14.3, 14.1HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3324.

[0650] 8-(Hexanoyloxy)octadecanoic acid

[0651] This compound was prepared according to the General procedure A.

[0652] 61% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.89-4.83 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.48 (m, 8 H), 1.33-1.25 (m, 26 H), 0.91-0.86 (m, 6 H).

[0653] 13C NMR (100 MHz, CDCl3) δ 179.3, 173.9, 74.1, 34.8, 34.3, 34.2, 34.0, 32.1, 31.5, 29.8, 29.7, 29.5, 29.3, 29.1, 25.5, 25.3, 25.0, 24.7, 22.8, 22.5, 14.3, 14.1.

[0654] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3321.

[0655] 9-(Hexanoyloxy)octadecanoic acid

[0656] This compound was prepared according to the General procedure A.

[0657] 36% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.59 (m, 4 H), 1.51-1.48 (m, 4 H), 1.33-1.26 (m, 26 H), 0.91- 0.86 (m, 6 H).

[0658] 13C NMR (100 MHz, CDCl3) δ 179.4, 173.9, 74.2, 34.9, 34.3, 34.27, 34.0, 32.0, 31.5, 29.7, 29.5, 29.3, 29.1, 25.5, 25.4, 25.0, 24.8, 22.8, 22.5, 14.3, 14.1.

[0659] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3318.

[0660] 10-(Hexanoyloxy)octadecanoic acid

[0661]

[0662] This compound was prepared according to the General procedure A.

[0663] 61% yield for 4 stepsXH NMR (400 MHz, CDCh) 54.89-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.59 (m, 4 H), 1.53-1.48 (m, 4 H), 1.32-1.26 (m, 26 H), 0.91- 0.86 (m, 6 H).

[0664] 13C NMR (100 MHz, CDCl3) δ 179.6, 173.9, 74.2, 34.9, 34.3, 34.1, 32.0, 31.5, 29.7, 29.64, 29.58, 29.44, 29.37, 29.3, 29.2, 25.5, 25.4, 25.0, 24.8, 22.8, 22.5, 14.2, 14.1.

[0665] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3315.

[0666] 11-(Hexanoyloxy)octadecanoic acid

[0667] This compound was prepared according to the General procedure A.

[0668] 40% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1 H), 2.37-2.26 (m, 4 H), 1.67-1.48 (m, 8 H), 1.33-1.26 (m, 26 H), 0.91-0.86 (m, 6 H).

[0669] 13C NMR (100 MHz, CDCl3) δ 178.6, 173.9, 74.2, 34.9, 34.3, 34.0, 31.9, 31.5, 29.64, 29.59, 29.5, 29.4, 29.3, 29.2, 25.5, 25.0, 24.9, 22.8, 22.5, 14.2, 14.1.

[0670] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3318.

[0671] 12-(Hexanoyloxy)octadecanoic acid D

[0672] This compound was prepared according to the General procedure A.

[0673] 70% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.59 (m, 4 H), 1.53-1.48 (m, 4 H), 1.34-1.26 (m, 26 H), 0.91- 0.86 (m, 6 H).

[0674] 13C NMR (100 MHz, CDCl3) δ 179.6, 173.9, 74.3, 34.9, 34.3, 34.1, 31.9, 31.5, 29.64, 29.63, 29.6, 29.5, 29.3, 29.2, 25.5, 25.4, 25.0, 24.8, 22.7, 22.5, 14.2, 14.1.

[0675] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3322.

[0676] 13-(Hexanoyloxy)octadecanoic acid

[0677]

[0678] This compound was prepared according to the General procedure A.

[0679] 60% yield for 4 steps

[0680] 1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1 H), 2.37-2.33 (m, 2 H), 2.30-2.25 (m, 2 H), 1.64-1.50 (m, 8 H), 1.322-1.26 (m, 26 H), 0.91-0.86 (m, 6 H).

[0681] 13C NMR (100 MHz, CDCl3) δ 179.1, 174.0, 74.3, 34.9, 34.30, 34.26, 34.1, 31.9, 31.5, 29.7, 29.5, 29.4, 29.2, 25.5, 25.1, 25.0, 24.9, 22.7, 22.5, 14.14, 14.06.

[0682] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3323.

[0683] 14-(Hexanoyloxy)octadecanoic acid

[0684]

[0685] This compound was prepared according to the General procedure A.

[0686] 52% yield for 4 steps

[0687] 1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.49 (m, 8 H), 1.34-1.24 (m, 26 H), 0.91-0.85 (m, 6 H).

[0688] 13C NMR (100 MHz, CDCl3) δ 179.4, 173.9, 74.3, 34.9, 34.3, 34.1, 34.0, 31.5, 29.7, 29.5, 29.4, 29.2, 27.6, 25.5, 25.0, 24.8, 22.8, 22.5, 14.2, 14.1.

[0689] HRMS (ESI): [M-H]⁻ calcd for C24H45O4397.3318, found 397.3327.

[0690] 15-(Hexanoyloxy)octadecanoic acid

[0691]

[0692] This compound was prepared according to the General procedure A.

[0693] 49% yield for 4 steps

[0694] 1H NMR (400 MHz, CDCl3) δ 4.92-4.86 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.59 (m, 4 H), 1.54-1.44 (m, 4 H), 1.37-1.25 (m, 26 H), 0.92-0.88 (m, 6 H).

[0695] 13C NMR (100 MHz, CDCl3) δ 179.6, 173.9, 74.0, 36.5, 34.9, 34.3, 34.1, 31.5, 29.8, 29.7, 29.6, 29.4, 29.2, 25.5, 25.0, 24.8, 22.5, 18.7, 14.2, 14.1.

[0696] HRMS (ESI): [M-H]⁻ calcd for C24H45O4397.3318, found 397.3318.

[0697] 16-(Hexanoyloxy)octadecanoic acidThis compound was prepared according to the General procedure A.

[0698] 55% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.85-4.78 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.50 (m, 8 H), 1.32-1.25 (m, 26 H), 0.91-0.85 (m, 6 H).

[0699] 13C NMR (100 MHz, CDCl3) δ 179.5, 174.0, 75.4, 34.9, 34.1, 33.8, 31.5, 29.8, 29.7, 29.6, 29.4, 29.2, 27.1, 25.5, 25.0, 24.8, 22.5, 14.1, 9.7.

[0700] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3324.

[0701] 17-(Hexanoyloxy)octadecanoic acid

[0702] This compound was prepared according to the General procedure A.

[0703] 34% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.94-4.86 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.27 (t, J = 7.6 Hz, 2 H), 1.64-1.41 (m, 8 H), 1.34-1.25 (m, 26 H), 1.19 (d, J = 6.0 Hz, 3 H), 0.92-0.88 (t, J = 6.4 Hz, 3 H).

[0704] 13C NMR (100 MHz, CDCl3) δ 179.7, 173.8, 70.9, 36.1, 34.9, 34.1, 31.5, 31.4, 29.8, 29.7, 29.60, 29.58, 29.4, 29.2, 25.6, 24.9, 24.8, 24.5, 22.5, 22.4, 20.2, 14.1, 14.0.

[0705] HRMS (ESI): [M-H] ’ calcd for C24H45O4397.3318, found 397.3323.

[0706] 2-(Palmitoyloxy)octadecanoic acid

[0707] This compound was prepared according to the General procedure A.

[0708] 35% yield for 4 steps

[0709]

[0710] 1H NMR (400 MHz, CDCl3) δ 5.01 (t, J = 6.4 Hz, 1 H), 2.39 (t, J = 7.6 Hz, 2 H), 1.89- 1.83 (m, 2 H), 1.67-1.61 (m, 2 H), 1.44-1.26 (m, 52 H), 0.90-0.86 (m, 6 H).

[0711] 13C NMR (100 MHz, CDCl3) δ 175.8, 173.6, 71.8, 34.1, 32.1, 31.1, 29.9, 29.82, 29.78, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2, 25.3, 25.0, 22.8, 14.3.

[0712] HRMS (ESI): [M-H]⁻ calcd for C34H65O4537.4883, found 537.4887.3-(Palmitoyloxy)octadecanoic acid

[0713] M.... X..

[0714] This compound was prepared according to the General procedure A.

[0715] 53% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 5.28-5.18 (m, 1 H), 2.63-2.53 (m, 2 H), 2.35 (t, J = 7.6 Hz, 1 H), 2.28 (t, J = 7.6 Hz, 1 H), 1.65-1.56 (m, 4 H), 1.32-1.26 (m, 50 H), 0.90-0.86 (m, 6 H).

[0716] 13C NMR (100 MHz, CDCl3) δ 179.3, 173.4, 70.1, 39.0, 34.6, 34.1, 34.0, 32.1, 29.9, 29.8, 29.74, 29.7, 29.64, 29.59, 29.52, 29.44, 29.39, 29.3, 29.2, 25.3, 25.2, 24.8, 22.9, 14.3.

[0717] HRMS (ESI): [M-H] ’ calcd for C34H65O4537.4883, found 537.4879.

[0718] 5-( Pal m itoyl oxy ) octadecanoi c aci d

[0719] This compound was prepared according to the General procedure A.

[0720] 44% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.86 (m, 1 H), 2.38-2.27 (m, 4 H), 1.67-1.51 (m, 8 H), 1.32-1.26 (m, 46 H), 0.89-0.86 (m, 6 H).

[0721] 13C NMR (100 MHz, CDCl3) δ 178.6, 173.9, 73.5, 34.8, 34.2, 33.7, 33.5, 32.1, 29.9, 29.8, 29.7, 29.69, 29.65, 29.5, 29.45, 29.4, 25.4, 25.3, 22.8, 20.6, 14.3.

[0722] HRMS (ESI): [M-H] ’ calcd for C34H65O4537.4883, found 537.4886.

[0723] 6-( Pal m itoyl oxy ) octadecanoi c aci d

[0724] . V.. ©

[0725]

[0726] "'■vr This compound was prepared according to the General procedure A.

[0727] 43% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.48 (m, 8 H), 1.34-1.26 (m, 46 H), 0.90-0.86 (m, 6 H).13C NMR (100 MHz, CDCl3) δ 178.9, 173.9, 73.8, 34.9, 34.3, 33.9, 33.87, 32.1, 29.9, 29.8, 29.7, 29.67, 29.51, 29.46, 29.4, 25.5, 25.3, 25.0, 24.7, 22.8, 14.3.

[0728] HRMS (ESI): [M-H] ’ calcd for C34H65O4 537.4883, found 537.4891.

[0729] 7-( Pal m itoyl oxy ) octadecanoi c aci d Q

[0730] Me'"'' O

[0731] This compound was prepared according to the General procedure A.

[0732] 58% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.48 (m, 8 H), 1.34-1.25 (m, 46 H), 0.90-0.86 (m, 6 H).

[0733] 13C NMR (100 MHz, CDCl3) δ 179.2, 173.9, 74.0, 34.9, 34.3, 34.1, 34.0, 32.1, 29.9, 29.8, 29.74, 29.69, 29.51, 29.46, 29.4, 29.1, 25.5, 25.3, 25.1, 24.7, 22.8, 14.3.

[0734] HRMS (ESI): [M-H] ’ calcd for C34H65O4 537.4883, found 537.4894.

[0735] 8-( Pal m itoyl oxy ) octadecanoi c aci d..,xW ‘Vv.y'' i

[0736] This compound was prepared according to the General procedure A.

[0737] 68% yield for 4 steps1H NMR (400 MHz, CDCl3) δ 4.89-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.64-1.48 (m, 8 H), 1.33-1.26 (m, 46 H), 0.90-0.86 (m, 6 H).

[0738] 13C NMR (100 MHz, CDCl3) δ 179.3, 173.9, 74.1, 34.9, 34.3, 34.2, 34.0, 32.1, 29.9, 29.8, 29.76, 29.7, 29.5, 29.46, 29.4, 29.3, 29.1, 25.5, 25.3, 25.26, 24.7, 22.8, 14.3.

[0739] HRMS (ESI): [M-H]⁻ calcd for C34H65O4537.4883, found 537.4888.

[0740] 9-(Palmitoyloxy)octadecanoic acid

[0741]

[0742] This compound was prepared according to the General procedure A.

[0743] 50% yield for 4 stepsXH NMR (400 MHz, CDCh) 54.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.58 (m, 4 H), 1.53-1.48 (m, 4 H), 1.30-1.26 (m, 46 H), 0.91-0.86 (m, 6 H).

[0744] 13C NMR (100 MHz, CDCh) 5179.2, 173.9, 74.2, 34.9, 34.3, 34.28, 34.0, 32.1, 32.0, 29.9, 29.8, 29.78, 29.6, 29.5, 29.46, 29.35, 29.3, 29.1, 25.5, 25.3, 24.8, 22.8, 14.3.

[0745] HRMS (ESI): [M-H] ’ calcd for C34H65O4537.4883, found 537.4879.

[0746] 10-(Palmitoyloxy)octadecanoic acid

[0747]

[0748] This compound was prepared according to the General procedure A.

[0749] 55% yield for 4 steps

[0750] XH NMR (400 MHz, CDCh) 64.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.58 (m, 4 H), 1.53-1.48 (m, 4 H), 1.32-1.26 (m, 46 H), 0.90-0.86 (m, 6 H).

[0751] 13C NMR (100 MHz, CDCh) 3 179.2, 173.9, 74.2, 34.9, 34.3, 34.0, 32.1, 32.0, 29.9, 29.8, 29.77, 29.69, 29.66, 29.59, 29.51, 29.46, 29.39, 29.35, 29.3, 29.2, 25.5, 25.4, 25.3, 24.8, 22.9,

[0752] 22.8, 14.3.

[0753] HRMS (ESI): [M-H] - calcd for C34H65O4537.4883, found 537.4886.

[0754] ll-(Palmitoyloxy)octadecanoic acid

[0755]

[0756] This compound was prepared according to the General procedure A.

[0757] 60% yield for 4 steps

[0758] XH NMR (400 MHz, CDCh) 64.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.48 (m, 8 H), 1.31-1.26 (m, 46 H), 0.90-0.86 (m, 6 H).

[0759] 13C NMR (100 MHz, CDCh) 3 179.3, 173.9, 74.2, 34.9, 34.3, 34.1, 32.1, 31.9, 29.85, 29.81, 29.77, 29.65, 29.51, 29.47, 29.4, 29.2, 25.5, 25.3, 24.8, 22.84, 22.78, 14.3, 14.2.

[0760] HRMS (ESI): [M-H] - calcd for C34H65O4537.4883, found 537.4879.

[0761] 12-(Palmitoyloxy)octadecanoic acido

[0762] ..-x >x... -x..-x A.

[0763] O -x -X " X x.x -X.-X

[0764]

[0765] |

[0766] This compound was prepared according to the General procedure A.

[0767] 67% yield for 4 steps

[0768] XH NMR (400 MHz, CDCI3) 64.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.58 (m, 4 H), 1.53-1.48 (m, 4 H), 1.34-1.26 (m, 46 H), 0.91-0.86 (m, 6 H).

[0769] 13C NMR (100 MHz, CDCI3) 3 179.4, 174.0, 74.3, 34.9, 34.3, 34.1, 31.9, 31.5, 32.1, 32.0, 29.9, 29.81, 29.77, 29.66, 29.64, 29.62, 29.52, 29.5, 29.4, 29.2, 25.5, 25.43, 25.35, 24.8, 22.8, 22.7, 14.3, 14.2.

[0770] HRMS (ESI): [M-H] - calcd for C34H65O4537.4883, found 537.4904.

[0771] 13-(Palmitoyloxy)octadecanoic acid

[0772]

[0773] This compound was prepared according to the General procedure A.

[0774] 52% yield for 4 steps

[0775] XH NMR (400 MHz, CDCI3) 54.89-4.85 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.26 (t, J = 7.6 Hz, 2 H), 1.67-1.48 (m, 8 H), 1.34-1.21 (m, 46 H), 0.91-0.86 (m, 6 H).

[0776] 13C NMR (100 MHz, CDCI3) 5 179.2, 174.0, 74.3, 34.9, 34.31, 34.27, 34.0, 32.1, 31.9, 29.9, 29.8, 29.77, 29.67, 29.55, 29.51, 29.46, 29.35, 29.2, 25.5, 25.3, 25.1, 24.8, 22.8, 22.7, 14.3, 14.2.

[0777] HRMS (ESI): [M-H] ■ calcd for C34H65O4537.4883, found 537.4883.

[0778] 14-(Palmitoyloxy)octadecanoic acid

[0779]

[0780] This compound was prepared according to the General procedure A.

[0781] 64% yield for 4 steps

[0782] XH NMR (400 MHz, CDCI3) 64.90-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.49 (m, 8 H), 1.34-1.25 (m, 46 H), 0.90-0.86 (m, 6 H).13C NMR (100 MHz, CDCh) 6 179.4, 174.0, 74.2, 34.9, 34.3, 34.1, 34.0, 32.1, 29.85, 29.81, 29.76, 29.70, 29.66, 29.57, 29.51, 29.46, 29.38, 29.34, 29.21, 27.6, 25.5, 25.3, 24.8, 22.8, 22.7, 14.3, 14.2.

[0783] HRMS (ESI): [M-H]⁻ calcd for C34H65O4537.4883, found 537.4894.

[0784] 15-(Palmitoyloxy)octadecanoic acid

[0785]

[0786] This compound was prepared according to the General procedure A.

[0787] 56% yield for 4 steps

[0788] 1H NMR (400 MHz, CDCl3) δ 4.92-4.86 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.57 (m, 4 H), 1.54-1.46 (m, 4 H), 1.37-1.26 (m, 46 H), 0.92-0.86 (m, 6 H).

[0789] 13C NMR (100 MHz, CDCh) 6 179.9, 173.9, 74.0, 36.5, 34.9, 34.3, 34.2, 32.1, 29.84, 29.81, 29.76, 29.72, 29.69, 29.65, 29.58, 29.51, 29.44, 29.38, 29.3, 29.2, 25.5, 25.3, 24.8, 22.8, 18.7, 14.3, 14.1.

[0790] HRMS (ESI): [M-H] ' calcd for C34H65O4537.4883, found 537.4889.

[0791] 16-(Palmitoyloxy)octadecanoic acid

[0792] O

[0793]

[0794] This compound was prepared according to the General procedure A.

[0795] 51% yield for 4 steps

[0796] 3H NMR (400 MHz, CDCh) 64.84-4.78 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.51 (m, 8 H), 1.35-1.25 (m, 46 H), 0.89-0.85 (m, 6 H).

[0797] 13C NMR (100 MHz, CDCh) 6179.5, 174.0, 75.4, 34.9, 34.1, 33.8, 32.1, 29.84, 29.80, 29.73, 29.71, 29.65, 29.58, 29.51, 29.45, 29.38, 29.34, 29.21, 27.1, 25.5, 25.4, 24.8, 22.8, 14.3, 9.8.

[0798] HRMS (ESI): [M-H] - calcd for C34H65O4537.4883, found 537.4879.

[0799] 17-(Palmitoyloxy)octadecanoic acidMe"'

[0800] This compound was prepared according to the General procedure A.

[0801] 65% yield for 4 stepsXH NMR (400 MHz, CDCI3) 54.94-4.86 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.26 (t, J = 7.6 Hz, 2 H), 1.67-1.25 (m, 54 H), 1.19 (d, J = 6.4 Hz, 3 H), 0.88 (t, J = 6.4 Hz, 3 H).

[0802] X3C NMR (100 MHz, CDCI3) 3 179.1, 173.8, 70.9, 36.1, 34.9, 34.1, 32.1, 29.8, 29.7, 29.64, 29.60, 29.50, 29.44, 29.39, 29.3, 29.2, 25.6, 25.3, 24.9, 22.8, 20.2, 14.3.

[0803] HRMS (ESI): [M-H] - calcd for C34H65O4537.4883, found 537.4902.

[0804] 2-(Stearoyloxy)octadecanoic acid

[0805] This compound was prepared according to the General procedure A.

[0806] 31% yield for 4 stepsXH NMR (400 MHz, CDCI3) 55.01 (t, J = 6.4 Hz, 1 H), 2.39 (t, J = 7.6 Hz, 2 H), 1.89- 1.83 (m, 2 H), 1.69-1.61 (m, 2 H), 1.44-1.22 (m, 56 H), 0.90-0.86 (m, 6 H).

[0807] 13C NMR (100 MHz, CDCI3) 6 175.9, 173.6, 71.8, 34.1, 32.1, 31.1, 29.9, 29.8, 29.77, 29.69, 29.6, 29.5, 29.4, 29.3, 29.2, 25.3, 25.0, 22.8, 14.3.

[0808] HRMS (ESI): [M-H] ' calcd for C36H69O4565.5196, found 565.5197.

[0809] 3-(Stearoyloxy)octadecanoic acid

[0810] This compound was prepared according to the General procedure A.

[0811] 44% yield for 4 steps

[0812]

[0813] XH NMR (400 MHz, CDCI3) 65.30-5.18 (m, 1 H), 2.66-2.56 (m, 4 H), 1.67-1.56 (m, 4 H), 1.34-1.21 (m, 54 H), 0.90-0.86 (m, 6 H).

[0814] X3C NMR (100 MHz, CDCI3) 6 175.3, 173.4, 70.2, 38.9, 34.6, 34.1, 29.9, 29.7, 29.64, 29.59, 29.52, 29.45, 29.4, 29.3, 29.2, 25.3, 25.2, 24.8, 22.9, 14.3.HRMS (ESI): [M-H] ’ calcd for C35H69O4565.5196, found 565.5207.

[0815] 5-(Stearoyloxy)octadecanoic acid

[0816] This compound was prepared according to the General procedure A.

[0817] 40% yield for 4 stepsXH NMR (400 MHz, CDCh) 64.92-4.85 (m, 1 H), 2.38-2.26 (m, 4 H), 1.67-1.49 (m, 8 H), 1.29-1.25 (m, 50 H), 0.90-0.86 (m, 6 H).

[0818] 13C NMR (100 MHz, CDCh) 3 178.2, 173.9, 73.5, 34.8, 34.2, 33.6, 33.5, 32.1, 29.9, 29.8, 29.73, 29.69, 29.65, 29.51, 29.46, 29.4, 25.4, 25.3, 22.8, 20.6, 14.3.

[0819] HRMS (ESI): [M-H] ’ calcd for C35H69O4565.5196, found 565.5211.

[0820] 6-(Stearoyloxy)octadecanoic acid

[0821] This compound was prepared according to the General procedure A.

[0822] 70% yield for 4 stepsXH NMR (400 MHz, CDCh) 54.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.26 (t, J = 7.6 Hz, 2 H), 1.66-1.48 (m, 8 H), 1.32-1.25 (m, 50 H), 0.90-0.86 (m, 6 H).

[0823] 13C NMR (100 MHz, CDCh) 5 178.8, 173.9, 73.8, 34.9, 34.3, 33.9, 33.86, 32.1, 29.9, 29.8, 29.73, 29.70, 29.67, 29.5, 29.46, 29.4, 25.5, 25.3, 25.0, 24.7, 22.8, 14.3.

[0824] HRMS (ESI): [M-H] ■ calcd for C36H69O4565.5196, found 565.5212.

[0825] 7-(Stearoyloxy)octadecanoic acid

[0826]

[0827] This compound was prepared according to the General procedure A.

[0828] 72% yield for 4 steps

[0829] XH NMR (400 MHz, CDCh) 54.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.48 (m, 8 H), 1.34-1.25 (m, 50 H), 0.90-0.86 (m, 6 H).13C NMR (100 MHz, CDCh) 6 179.0, 173.9, 74.0, 34.9, 34.3, 34.1, 33.9, 32.1, 29.9, 29.8, 29.79, 29.7, 29.68, 29.5, 29.46, 29.4, 29.1, 25.5, 25.3, 25.1, 24.7, 22.8, 14.3.

[0830] HRMS (ESI): [M-H] ’ calcd for C36H69O4565.5196, found 565.5192.

[0831] 8-(Stearoyloxy)octadecanoic acid

[0832] o This compound was prepared according to the General procedure A.

[0833] 49% yield for 4 stepsXH NMR (400 MHz, CDCh) 54.89-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.64-1.48 (m, 8 H), 1.32-1.25 (m, 50 H), 0.90-0.86 (m, 6 H).

[0834] 13C NMR (100 MHz, CDCh) 6 179.1, 173.9, 74.1, 34.9, 34.3, 34.2, 34.0, 32.1, 29.9, 29.82, 29.78, 29.76, 29.74, 29.69, 29.51, 29.49, 29.46, 29.4, 29.3, 29.1, 25.5, 25.33, 25.27, 24.7, 22.8, 14.3.

[0835] HRMS (ESI): [M-H] ’ calcd for C36H69O4 565.5196, found 565.5200.

[0836] 9-(Stearoyloxy)octadecanoic acid

[0837] This compound was prepared according to the General procedure A.

[0838] 50% yield for 4 stepsXH NMR (400 MHz, CDCh) 64.89-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.58 (m, 4 H), 1.51-1.48 (m, 4 H), 1.32-1.25 (m, 50 H), 0.90- 0.86 (m, 6 H).

[0839] 13C NMR (100 MHz, CDCh) 6 179.5, 173.9, 74.2, 34.9, 34.3, 34.27, 34.1, 32.1, 32.0, 29.9, 29.8, 29.78, 29.69, 29.5, 29.46, 29.4, 29.3, 29.1, 25.5, 25.4, 25.3, 24.8, 22.8, 14.3.

[0840] HRMS (ESI): [M-H] ’ calcd for C36H69O4565.5196, found 565.5195.

[0841] 10-(Stearoyloxy)octadecanoic acid

[0842]

[0843] This compound was prepared according to the General procedure A.

[0844] 63% yield for 4 steps

[0845] XH NMR (400 MHz, CDCh) 64.90-4.83 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.66-1.58 (m, 4 H), 1.53-1.48 (m, 4 H), 1.30-1.26 (m, 50 H), 0.90-0.86 (m, 6 H).

[0846] 13C NMR (100 MHz, CDCh) 5 179.2, 173.9, 74.2, 34.9, 34.3, 34.0, 32.1, 32.0, 29.85, 29.81, 29.78, 29.69, 29.67, 29.59, 29.52, 29.46, 29.39, 29.35, 29.3, 29.2, 25.5, 25.4, 25.35, 24.8, 22.8, 14.3.

[0847] HRMS (ESI): [M-H] ’ calcd for C36H69O4 565.5196, found 565.5200.

[0848] 1 l-(Stearoyloxy)octadecanoic add

[0849]

[0850] This compound was prepared according to the General procedure A.

[0851] 49% yield for 4 steps

[0852] 3H NMR (400 MHz, CDCh) 64.90-4.83 (m, 1 H), 2.36-2.26 (m, 4 H), 1.67-1.48 (m, 8 H), 1.35-1.26 (m, 50 H), 0.89-0.86 (m, 6 H).

[0853] 13C NMR (100 MHz, CDCh) 3 179.1, 174.0, 74.2, 34.9, 34.3, 34.0, 32.1, 31.9, 29.9, 29.81, 29.78, 29.66, 29.61, 29.51, 29.47, 29.4, 29.2, 25.5, 25.4, 24.8, 22.84, 22.79, 14.3.

[0854] HRMS (ESI): [M-H] ’ calcd for C36H69O4 565.5196, found 565.5197.

[0855] 12-(Stearoyloxy)octadecanoic acid

[0856]

[0857] This compound was prepared according to the General procedure A.

[0858] 38% yield for 4 steps

[0859] XH NMR (400 MHz, CDCh) 64.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.58 (m, 4 H), 1.53-1.48 (m, 4 H), 1.33-1.26 (m, 50 H), 0.89-0.86 (m, 6 H).

[0860] 13C NMR (100 MHz, CDCh) 3 179.3, 174.0, 74.3, 34.9, 34.3, 34.1, 32.1, 31.9, 29.9, 29.81, 29.77, 29.67, 29.64, 29.62, 29.52, 29.5, 29.4, 29.2, 25.5, 25.43, 25.35, 24.8, 22.8, 22.7, 14.3, 14.2.

[0861] HRMS (ESI): [M-H] ■ calcd for C36H69O4565.5196, found 565.5204.13-(Stearoyloxy)octadecanoic acid

[0862]

[0863] This compound was prepared according to the General procedure A.

[0864] 66% yield for 4 steps

[0865] XH NMR (400 MHz, CDCh) 64.90-4.84 (m, 1 H), 2.36-2.26 (m, 4 H), 1.65-1.48 (m, 8 H), 1.35-1.26 (m, 50 H), 0.90-0.86 (m, 6 H).

[0866] 13C NMR (100 MHz, CDCh) 6 179.4, 174.0, 74.3, 34.9, 34.31, 34.26, 34.1, 32.1, 31.9, 29.9, 29.8, 29.77, 29.67, 29.55, 29.51, 29.46, 29.36, 29.2, 25.5, 25.3, 25.1, 24.9, 22.8, 22.7, 14.3, 14.2.

[0867] HRMS (ESI): [M-H] ’ calcd for C36H69O4 565.5196, found 565.5209.

[0868] 14-(Stearoyloxy)octadecanoic acid

[0869]

[0870] This compound was prepared according to the General procedure A.

[0871] 36% yield for 4 steps

[0872] XH NMR (400 MHz, CDCh) 5 4.90-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.49 (m, 8 H), 1.32-1.25 (m, 50 H), 0.90-0.86 (m, 6 H).

[0873] 13C NMR (100 MHz, CDCh) 6 179.3, 174.0, 74.2, 34.9, 34.3, 34.1, 34.0, 32.1, 29.85, 29.81, 29.77, 29.70, 29.57, 29.51, 29.46, 29.35, 29.21, 27.65, 25.5, 25.3, 24.8, 22.8, 22.7, 14.3, 14.2.

[0874] HRMS (ESI): [M-H] ■ calcd for C36H69O4 565.5196, found 565.5200.

[0875] 15-(Stearoyloxy)octadecanoic acid

[0876]

[0877] This compound was prepared according to the General procedure A.

[0878] 76% yield for 4 stepsXH NMR (400 MHz, CDCh) 54.92-4.86 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.60 (m, 4 H), 1.54-1.46 (m, 4 H), 1.35-1.26 (m, 50 H), 0.92-0.86 (m, 6 H).

[0879] 13C NMR (100 MHz, CDCh) 5179.8, 174.0, 74.0, 36.5, 34.9, 34.3, 34.1, 32.1, 29.85, 29.81, 29.76, 29.72, 29.70, 29.65, 29.58, 29.51, 29.45, 29.39, 29.3, 29.2, 25.5, 25.3, 24.8, 22.8, 18.7, 14.3, 14.1.

[0880] HRMS (ESI): [M-H] ’ calcd for C36H69O4565.5196, found 565.5190.

[0881] 16-(Stearoyloxy)octadecanoic acid

[0882]

[0883] This compound was prepared according to the General procedure A.

[0884] 53% yield for 4 steps

[0885] XH NMR (400 MHz, CDCh) S 4.85-4.78 (m, 1 H), 2.35 (t, J = 7.6, 2 H), 2.29 (t, J = 7.6, 2 H), 1.65-1.50 (m, 8 H), 1.30-1.25 (m, 50 H), 0.89-0.85 (m, 6 H).

[0886] 13C NMR (100 MHz, CDCh) 6179.6, 174.0, 75.4, 34.9, 34.1, 33.8, 32.1, 29.85, 29.80, 29.73, 29.70, 29.65, 29.58, 29.51, 29.45, 29.39, 29.3, 29.2, 27.1, 25.5, 25.4, 24.8, 22.8, 14.3, 9.8.

[0887] HRMS (ESI): [M-H] ' calcd for C36H69O4565.5196, found 565.5201.

[0888] 17-(Stearoyloxy)octadecanoic acid

[0889]

[0890] This compound was prepared according to the General procedure A.

[0891] 56% yield for 4 steps

[0892] XH NMR (400 MHz, CDCh) 64.94-4.86 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.26 (t, J = 7.6 Hz, 2 H), 1.67-1.53 (m, 4 H), 1.49-1.25 (m, 54 H), 1.19 (d, J = 6.4 Hz, 3 H), 0.88 (t, J = 6.4 Hz, 3 H).

[0893] X3C NMR (100 MHz, CDCh) 6179.4, 173.8, 70.9, 36.1, 34.9, 34.1, 32.1, 29.84, 29.81, 29.73, 29.64, 29.61, 29.58, 29.51, 29.44, 29.39, 29.3, 29.2, 25.6, 25.3, 24.8, 22.8, 20.2, 14.3.

[0894] HRMS (ESI): [M-H] ' calcd for C36H69O4 565.5196, found 565.5191.12-(decanoyloxy)octadecanoic acid

[0895]

[0896] This compound was prepared according to the General procedure A.

[0897] 61% yield for 4 steps

[0898] XH NMR (400 MHz, CDCh) 64.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.48 (m, 8 H), 1.33-1.26 (m, 34 H), 0.91-0.86 (m, 6 H).

[0899] 13C NMR (100 MHz, CDCh) 5 179.4, 174.0, 74.3, 34.9, 34.3, 34.1, 32.0, 31.9, 29.7, 29.6, 29.52, 29.46, 29.42, 29.35, 29.2, 25.5, 25.3, 24.8, 22.8, 22.7, 14.22, 14.21.

[0900] HRMS (ESI): [M-H] ■ calcd for C28H53O4453.3949, found 453.3949.

[0901] 12-(dodecanoyloxy)octadecanoic acid

[0902] o

[0903]

[0904] This compound was prepared according to the General procedure A.

[0905] 49% yield for 4 steps

[0906] 3H NMR (400 MHz, CDCh) 64.90-4.84 (m, 1 H), 2.355 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.48 (m, 8 H), 1.35-1.26 (m, 38 H), 0.90-0.86 (m, 6 H).

[0907] 13C NMR (100 MHz, CDCh) 3 179.3, 174.0, 74.3, 34.9, 34.3, 34.1, 32.1, 31.9, 29.8, 29.74, 29.66, 29.62, 29.52, 29.5, 29.4, 29.2, 25.5, 25.4, 25.3, 24.8, 22.8, 22.7, 14.26, 14.21.

[0908] HRMS (ESI): [M-H] ' calcd for C30H57O4481.4262, found 4881.4262.

[0909] 12-(Tetradecanoyloxy)octadecanoic acid

[0910]

[0911] This compound was prepared according to the General procedure A.

[0912] 78% yield for 4 steps

[0913] 1H NMR (400 MHz, CDCl3) δ 4.91-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.48 (m, 8 H), 1.30-1.26 (m, 42 H), 0.91-0.86 (m, 6 H).13C NMR (100 MHz, CDCl₃) δ 179.4, 174.0, 74.3, 34.9, 34.3, 34.1, 32.1, 31.9, 29.81, 29.77, 29.66, 29.62, 29.51, 29.46, 29.35, 29.19, 25.46, 25.42, 25.3, 24.8, 22.8, 22.7, 14.3, 14.2.

[0914] HRMS (ESI): [M-H] ' calcd for C₃₂H₆₁O₄ 509.4570, found 509.4577.

[0915] 12-(Palmitoyloxy)hexadecanoic acid

[0916]

[0917] This compound was prepared according to the General procedure A.

[0918] 55% yield for 4 steps

[0919] 1H NMR (400 MHz, CDCl₃) δ 4.90-4.84 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.48 (m, 8 H), 1.32-1.26 (m, 42 H), 0.90-0.86 (m, 6 H).

[0920] 13C NMR (100 MHz, CDCl₃) δ 179.5, 174.0, 74.3, 34.9, 34.3, 34.1, 34.0, 32.1, 32.0, 29.84, 29.80, 29.77, 29.66, 29.62, 29.51, 29.46, 29.39, 29.34, 29.19, 25.5, 25.3, 24.8, 22.8, 14.3, 14.2.

[0921] HRMS (ESI): [M-H] ’ calcd for C₃₂H₆₁O₄ 509.4570, found 509.4576.

[0922] 12-(Palmitoyloxy)icosanoic acid

[0923] o

[0924]

[0925] This compound was prepared according to the General procedure A.

[0926] 66% yield for 4 steps

[0927] 1H NMR (400 MHz, CDCl₃) δ 4.90-4.84 (m, 1 H), 2.36 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.65-1.49 (m, 8 H), 1.32-1.21 (m, 50 H), 0.90-0.86 (m, 6 H).

[0928] 13C NMR (100 MHz, CDCl₃) δ 179.6, 174.0, 74.2, 34.9, 34.3, 34.1, 34.0, 32.1, 29.84, 29.81, 29.76, 29.65, 29.62, 29.52, 29.45, 29.35, 29.19, 27.6, 25.5, 25.3, 24.8, 22.84, 22.75, 14.3, 14.2.

[0929] HRMS (ESI): [M-H] ■ calcd for C₃₆H₆₉O₄ 565.5196, found 565.5208.

[0930] 12-(Icosanoyloxy)octadecanoic acidy-x / x / X / XX

[0931]

[0932] d

[0933] This compound was prepared according to the General procedure A.

[0934] 70% yield for 4 steps

[0935] 1H NMR (400 MHz, CDCl₃) δ 4.90-4.84 (m, 1 H), 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.48 (m, 8 H), 1.35-1.19 (m, 54 H), 0.90-0.86 (m, 6 H).

[0936] 13C NMR (100 MHz, CDCl₃) δ 179.2, 174.0, 74.3, 34.9, 34.3, 34.0, 32.1, 31.9, 29.85, 29.81, 29.78, 29.66, 29.64, 29.62, 29.52, 29.46, 29.35, 29.19, 25.43, 25.3, 24.8, 22.8, 22.7, 14.3, 14.2.

[0937] HRMS (ESI): [M-H] calcd for C₃₈H₇₃O₄ 593.5509, found 593.5521.

[0938] 12-(Tetradecanoyloxy)octadecanoic-12-d acid

[0939]

[0940] This compound was prepared according to the General procedure B.

[0941] 58% yield for 2 steps

[0942] 1H NMR (400 MHz, CDCl₃) δ 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.58 (m, 4 H), 1.52-1.48 (m, 4 H), 1.33-1.26 (m, 41 H), 0.90-0.86 (m, 6 H).

[0943] 13C NMR (100 MHz, CDCl₃) δ 179.1, 174.0, 2.35 (t, J = 7.6 Hz, 2 H), 34.9, 34.2, 34.0, 32.1, 31.9, 29.81, 29.78, 29.66, 29.62, 29.51, 29.46, 29.35, 29.19, 25.43, 25.39, 25.34, 24.8, 22.8, 22.7, 14.3, 14.2.

[0944] HRMS (ESI): [M-H] ■ calcd for C₃₂H₆₀DO₄ 510.4635, found 510.4640.

[0945] 12-(Stearoyloxy)octadecanoic-12-d acid

[0946]

[0947] This compound was prepared according to the General procedure B.

[0948] 62% yield for 2 steps

[0949] 1H NMR (400 MHz, CDCl₃) δ 2.35 (t, J = 7.6 Hz, 2 H), 2.28 (t, J = 7.6 Hz, 2 H), 1.67-1.58 (m, 4 H), 1.51-1.48 (m, 4 H), 1.30-1.26 (m, 49 H), 0.89-0.86 (m, 6 H).13C NMR (100 MHz, CDCl₃) δ 178.9, 174.0, 73.9 (t, J = 23 Hz, 3 C), 34.9, 34.2, 34.0, 32.1, 31.9, 29.9, 29.8, 29.77, 29.66, 29.62, 29.51, 29.46, 29.35, 29.19, 25.43, 25.39, 25.34, 24.8, 22.8, 22.7, 14.3, 14.2.

[0950] HRMS (ESI): [M-H] ' calcd for C₃₆H₆₉DO₄ 567.5352, found 567.5334.

[0951] 12-Tetradecanamidooctadecanoic acid

[0952] This compound was prepared according to the General procedure D.

[0953] 47% yield for 3 stepsXH NMR (400 MHz, CDCh) 6 5.13 (d, J = 9.2 Hz, 1 H), 3.94-3.87 (m, 1 H), 3.36-3.32 (m, 2 H), 2.19-2.15 (m, 2 H), 1.67 -1.25 (m, 50 H), 0.90-0.86 (m, 6 H).

[0954] 13C NMR (100 MHz, CDCl₃) δ 178.4, 173.1, 37.3, 35.5, 34.1, 34.1, 32.1, 31.9, 29.80, 29.69, 29.62, 29.53, 29.50, 29.3, 29.1, 29.0, 26.1, 26.0, 24.8, 22.8, 22.7, 14.3, 14.2.

[0955] HRMS (ESI): [M-H] ' calcd for C₃₂H₆₂NO₃ 508.4730, found 508.4738.

[0956] 12-Stearamidooctadecanoic acid

[0957] This compound was prepared according to the General procedure D.

[0958] 65% yield for 3 stepsXH NMR (400 MHz, CDCh) 6 5.14 (d, J = 9.2 Hz, 1 H), 3.93-3.87 (m, 1 H), 3.36-3.32 (m, 2 H), 2.19-2.15 (m, 2 H), 1.67 -1.15 (m, 58 H), 0.90-0.86 (m, 6 H).

[0959] 13C NMR (100 MHz, CDCl₃) δ 178.7, 173.1, 49.3, 37.3, 35.5, 32.1, 31.9, 29.84, 29.69, 29.64, 29.53, 29.50, 29.4, 29.2, 29.0, 26.1, 26.0, 24.9, 22.8, 22.7, 14.3, 14.2.

[0960] HRMS (ESI): [M-H] ’ calcd for C₃₆H₇₀NO₃ 564.5356, found 564.5364.

[0961] 12-((Dodecylsulfonyl)oxy)octadecanoic acid

[0962]

[0963] This compound was prepared according to the General procedure E.73% yield for 2 steps

[0964] XH NMR (400 MHz, CDCh) 64.74-4.68 (m, 1 H), 3.07-3.03 (m, 2 H), 2.37-2.33 (m, 2 H), 1.88-1.81 (m, 2 H), 1.70 -1.61 (m, 6 H), 1.42 -1.26 (m, 40 H), 0.90-0.87 (m, 6 H).

[0965] 13C NMR (100 MHz, CDCl₃) δ 179.0, 83.6, 51.7, 34.6, 32.1, 31.8, 29.74, 29.66, 29.59, 29.55, 29.51, 29.47, 29.42, 29.34, 29.22, 29.17, 28.4, 25.1, 25.0, 23.7, 22.8, 22.7, 14.3, 14.2.

[0966] HRMS (ESI): [M-H] ’ calcd for C₃₀H₅₉O₅S 531.4083, found 531.4084.

[0967] 12-((Hexadecylsulfonyl)oxy)octadecanoic acid

[0968]

[0969] This compound was prepared according to the General procedure E.

[0970] 66% yield for 2 steps

[0971] XH NMR (400 MHz, CDCh) 64.74-4.68 (m, 1 H), 3.07-3.03 (m, 2 H), 2.35 (t, J = 7.6 Hz, 2 H), 1.89 -1.81 (m, 2 H), 1.71 -1.59 (m, 6 H), 1.42 -1.26 (m, 48 H), 0.90-0.86 (m, 6 H).

[0972] 13C NMR (100 MHz, CDCl₃) δ 179.4, 83.6, 51.7, 34.6, 34.1, 32.1, 31.8, 29.83, 29.80, 29.75, 29.66, 29.59, 29.56, 29.54, 29.50, 29.43, 29.34, 29.22, 29.17, 28.4, 25.03, 25.00, 24.8, 23.7, 22.8, 22.7, 14.3, 14.2.

[0973] HRMS (ESI): [M-H] ■ calcd for C₃₄H₆₇O₅S 587.4709, found 587.4717.

[0974] 12-(((Dodecyloxy)carbonyl)oxy)icosanoic acid

[0975]

[0976] This compound was prepared according to the General procedure F.

[0977] 66% yield for 2 steps

[0978] XH NMR (400 MHz, CDCh) 64.71-4.65 (m, 1 H), 4.13-4.10 (m, 2 H), 2.37-2.33 (m, 2 H), 1.68-1.53 (m, 2 H), 1.32 -1.26 (m, 48 H), 0.90-0.86 (m, 6 H).

[0979] 13C NMR (100 MHz, CDCl₃) δ 179.3, 155.5, 79.0, 68.0, 34.3, 34.1, 32.1, 31.9, 29.84, 29.72, 29.66, 29.64, 29.61, 29.51, 29.40, 29.35, 29.33, 29.19, 28.87, 25.9, 25.4, 25.3, 24.8, 22.8, 22.7, 14.3, 14.2.

[0980] HRMS (ESI): [M-H] ’ calcd for C₃₁H₅₉O₅ 511.4362, found 511.4376.12-(((Hexadecyloxy)carbonyl)oxy)octadecanoic acid 0

[0981] V

[0982] This compound was prepared according to the General procedure F.

[0983] 60% yield for 2 stepsXH NMR (400 MHz, CDCl₃) δ 4.71-4.67 (m, 1 H), 4.13-4.10 (m, 2 H), 2.37-2.33 (m, 2 H), 1.68-1.53 (m, 8 H), 1.32 -1.26 (m, 48 H), 0.90-0.86 (m, 6 H).

[0984] 13C NMR (100 MHz, CDCl₃) δ 179.2, 155.5, 79.0, 68.0, 34.3, 34.0, 32.1, 31.9, 29.78, 29.71, 29.66, 29.61, 29.51, 29.40, 29.34, 29.19, 28.88, 25.9, 25.36, 25.32, 24.8, 22.8, 22.7, 14.3, 14.2.

[0985] HRMS (ESI): [M-H] calcd for C₃₅H₆₇O₅ 567.4989, found 567.4985.

[0986] 12-(5-Dodecyl-1H-1,2,3-triazol-1-yl)octadecanoic acid

[0987] This compound was prepared according to the General procedure G.

[0988] 50% yield for 4 stepsXH NMR (400 MHz, CDCl₃) δ 7.20 (s, 1 H), 4.46-4.39 (m, 1 H), 2.73-2.70 (m, 2 H), 2.37-2.33 (m, 2 H), 1.85 -1.78 (m, 4 H), 1.68 -1.59 (m, 4 H), 1.34 -1.02 (m, 40 H), 0.90-0.83 (m, 6 H).

[0989] 13C NMR (100 MHz, CDCl₃) δ 178.1, 148.4, 118.7, 62.2, 35.93, 35.88, 34.1, 32.1, 31.7, 29.83, 29.79, 29.74, 29.57, 29.53, 29.51, 29.4, 29.3, 29.1, 29.0, 28.9, 26.1, 26.0, 25.9, 24.8, 22.8, 22.6, 14.3, 14.1.

[0990] HRMS (ESI): [M-H] - calcd for C₃₂H₆₀N₃O₂ 518.4686, found 518.4685.

[0991] 12-(5-Hexadecyl-1H-1,2,3-triazol-1-yl)octadecanoic acid

[0992] O

[0993]

[0994] This compound was prepared according to the General procedure G.

[0995] 50% yield for 4 steps1H NMR (400 MHz, CDCl₃) δ 7.20 (s, 1 H), 4.46-4.39 (m, 1 H), 2.74-2.70 (m, 2 H), 2.36-2.33 (m, 2 H), 1.85 -1.76 (m, 4 H), 1.68 -1.59 (m, 4 H), 1.34 -1.05 (m, 48 H), 0.90-0.83 (m, 6 H).

[0996] 13C NMR (100 MHz, CDCl₃) δ 178.7, 148.3, 118.7, 62.2, 35.90, 35.87, 34.2, 32.1, 31.7, 29.84, 29.80, 29.73, 29.56, 29.53, 29.50, 29.38, 29.34, 29.18, 29.16, 29.1, 26.03, 25.98, 25.03, 24.8, 22.8, 22.6, 14.3, 14.1.

[0997] HRMS (ESI): [M-H] ’ calcd for C₃₆H₆₈N₃O₂ 574.5312, found 574.5308.

[0998] 12-((2, 2, 3, 3, 4, 4, 5, 5, 6, 6, 7, 7, 8, 8, 9, 9, 10, 10, 11, 11, 12, 12, 13, 13, 14, 14,14-Heptacosafluorotetradecanoyl)oxy)octadecanoic acid

[0999]

[1000] This compound was prepared according to the General procedure A.

[1001] 30% yield for 4 steps

[1002] XH NMR (400 MHz, CDCl₃) δ 5.13-5.07 (m, 1 H), 2.36-2.33 (m, 2 H), 1.66 -1.61 (m, 4 H), 1.32-1.27 (m, 24 H), 0.89-0.86 (m, 3 H).

[1003] 13C NMR (100 MHz, CDCl₃) δ 179.9, 158.6, 158.3, 158.0, 130.6, 130.5, 130.4, 80.8, 62.2, 34.1, 33.7, 32.8, 31.7, 29.8, 29.7, 29.6, 29.5, 29.4, 29.33, 29.28, 29.22, 29.18, 29.1, 24.99, 24.95, 24.84, 24.80, 22.6, 14.2, 14.1.35.90, 35.87, 34.2, 32.1, 31.7, 29.84, 29.80, 29.73, 29.56, 29.53, 29.50, 29.38, 29.34, 29.18, 29.16, 29.1, 26.03, 25.98, 25.03, 24.8, 22.8, 22.6, 14.3, 14.1.

[1004] HRMS (ESI): [M-H] ■ calcd for C₃₂H₃₄F₂₇O₄ 995.2031, found 995.2021.

[1005] 12-(Tetradec-13-ynamido)octadecanoic acid

[1006]

[1007] To a solution of ethynyltrimethylsilane (2.0 equiv.) in dry THF (0.5 M), n-BuLi (2.0 equiv.) was added at -78 °C. After 15 minutes of stirring, a solution of 12-bromododecanoic acid (1.0 equiv.) in THF and HMPA was added to the mixture. The reaction was stirred at -78 °C for 1 hour and stirred for another 2h at room temperature. The mixture was quenched with NH4CI solution. The mixture was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EtOAc to give the 14-(trimethylsilyl)tetradec-13-ynoic acid.

[1008] To a solution of 14-(trimethylsilyl)tetradec-13-ynoic acid (1.0 equiv.) in dry DCM (1 M), methyl 12-aminooctadecanoate (2 equiv.), DCC (2.5 equiv.) and DMAP (10 mol%) were added at RT. The reaction was stirred at room temperature for 12 hours. The mixture was quenched with NH4CI, which was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using Hexane / EA to give the product.

[1009] The methyl 12-(tetradec-13-ynamido)octadecanoate (1 equiv.) was dissolved in MeOH: THF: H2O (1:2: 1) and cooled to 0 °C for 15 minutes before adding solid KOH (6.5 equiv.) in one portion. The reaction was allowed to warm to 23 °C and stirred for 24 hours. Excess hydroxide was neutralized with 1 N HCI. The resulting product was extracted with DCM (10 mL x 3), and the combined organic extracts were washed with brine (5 mL), dried over sodium sulfate, filtered, and concentrated. The crude products were subsequently purified by silica gel column chromatography, eluting with DCM / Methanol to yield products.

[1010] XH NMR (400 MHz, CDCl₃) δ 5.14 (d, J = 9.2 Hz, 1 H), 3.94-3.89 (m, 1 H), 2.34 (t, J = 7.6 Hz, 2 H), 2.20-2.15 (m, 4 H), 1.94 (t, J =2.0 Hz, 1 H), 1.65-1.25 (m, 46 H), 0.89- 0.86 (m, 3H).

[1011] 13C NMR (100 MHz, CDCl₃) δ 179.4, 173.1, 85.0, 68.2, 49.3, 37.3, 35.5, 34.2, 31.9, 29.6, 29.5, 29.4, 29.36, 29.22, 29.15, 29.0, 28.9, 29.6, 26.1, 26.0, 24.9, 24.8, 22.7, 18.5, 14.2.

[1012] HRMS (ESI): [M-H] ’ calcd for C₃₂H₅₈NO₃ 504.4422, found 504.4420.

[1013] 12-(12-Azidododecanamido)octadecanoic acid. a „.. Ix'-''.. *x"' Y’x""' x k Methyl 12-(12-bromododecanamido)octadecanoate (1 equiv.) and sodium azide (4

[1014]

[1015] equiv.) in DMF (1 M) were stirred at 100 °C for 12 hours. After quenching with water, the mixture was extracted twice with ethyl acetate. Combined organic layers werewashed (brine), dried (Na2SO4), filtered, and evaporated. Purification provided the azide intermediate.

[1016] The methyl 12-(12-azidododecanamido)octadecanoate (1 equiv.) was dissolved in THF: H2O (1:1) and cooled to 0 °C for 15 minutes before adding solid LIOH (6.5 equiv.) in one portion. The reaction was allowed to warm to 23 °C and stirred for 24 hours. Excess hydroxide was neutralized with 1 N HCI. The resulting product was extracted with DCM (10 mL x 3), and the combined organic extracts were washed with brine (5 mL), dried over sodium sulfate, filtered, and concentrated. The crude products were subsequently purified by silica gel column chromatography, eluting with DCM / Methanol to yield products.

[1017] XH NMR (400 MHz, CDCl₃) δ 5.10 (d, J = 9.2 Hz, 1 H), 3.93-3.89 (m, 1 H), 3.27-3.24 (m, 2 H), 2.36-2.32 (m, 2 H), 2.19-2.15 (m, 2 H), 1.65-1.26 (m, 46 H), 0.89-0.86 (m, 3H).

[1018] 13C NMR (100 MHz, CDCl₃) δ 177.3, 173.1, 51.6, 49.3, 37.3, 35.5, 33.9, 31.9, 29.9, 29.61, 29.55, 29.50, 29.41, 29.28, 29.18, 29.16, 28.98, 28.94, 28.86, 28.85, 26.1, 26.0, 25.9, 24.8, 22.7, 14.3, 14.2

[1019] HRMS (ESI): [M-H] ■ calcd for C₃₀H₅₇N₄O₃ 521.4426, found 521.4440.

[1020] 2-((5-((3aS,4S,6aR)-2-oxohexahydro-lH-thieno[3,4-d]imidazol-4-yl)pentanoyl)oxy)ethyl 12-tetradecanamidooctadecanoate

[1021]

[1022] To a solution of Biotin (1.0 equiv.) in dry DCM (1 M), Ethylene glycol (2 equiv.), DCC (2.5 equiv.) and DMAP (10 mol%) were added at RT. The reaction was stirred at room temperature for 12 hours. The mixture was quenched with NH4CI, which was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using DCM / MeOH to give the product.To a solution of above product (1.0 equiv.) in dry DCM (IM), 12-tetradecanamidooctadecanoic acid (1 equiv.), DCC (2.5 equiv.) and DMAP (10 mol%) were added at RT. The reaction was stirred at room temperature for 12 hours. The mixture was quenched with NH4CI, which was extracted with DCM (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified via silica gel column chromatography using DCM / MeOH to give the product

[1023] XH NMR (400 MHz, CDCl₃) δ 5.87 (s, 1 H), 5.53 (s, 1 H), 5.12 (d, J = 9.2 Hz, 1 H), 4.55-4.51 (m, 1 H), 4.32-4.29 (m, 4 H), 3.96-3.88 (m, 1 H), 3.69-3.64 (m, 1 H), 3.20-3.15 (m, 1 H), 2.95-2.90 (m, 1 H), 2.77-2.74 (m, 2 H), 2.45-2.41 (m, 2 H), 2.18-2.14 (m, 2 H), 2.19-2.15 (m, 2 H), 1.79-1.25 (m, 55 H), 0.90-0.86 (m, 6 H).

[1024] 13C NMR (1OO MHz, CDCl₃) δ 173.0, 167.9, 154.2, 65.7, 62.0, 60.4, 55.3, 49.3, 40.6, 37.3, 35.5, 35.0, 34.2, 32.8, 32.1, 31.9, 31.2, 30.7, 29.8, 29.69, 29.64, 29.53, 29.50, 29.43, 29.38, 29.33, 29.14, 29.1, 28.31, 28.27, 26.4, 26.1, 26.0, 25.6, 25.5, 25.2, 25.0, 24.9, 22.8, 22.7, 19.3, 14.3, 14.2, 13.9.

[1025] HRMS (ESI): [M+H]+calcd for C₄₄H₈₂N₃O₆S 780.5924, found 780.5917.

[1026] It will be appreciated that many further modifications and permutations of various aspects of the described embodiments are possible. Accordingly, the described aspects are intended to embrace all such alterations, modifications, and variations that fall within the spirit and scope of the appended claims.

[1027] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[1028] Throughout this specification and the claims which follow, unless the context requires otherwise, the phrase "consisting essentially of", and variations such as "consists essentially of" will be understood to indicate that the recited element(s) is / are essential i.e. necessary elements of the invention. The phrase allows for the presence of other non-recited elements which do not materially affect the characteristics of the invention but excludes additional unspecified elements which would affect the basic and novel characteristics of the method defined.

[1029] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as anacknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Claims

Claims1. A method of synthesising a compound of Formula (I), or a salt, solvate, stereoisomer, prodrug or derivative thereof, comprising:~^H? L Oxw™ (!)whereinL is a linker selected from oxylene, oxyacylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;m is an integer selected from 0 to 20;n is an integer selected from 0 to 20;i is an integer selected from 0 to 20;a) conjugating an alkyl moiety via linker to the hydroxy-alkyl ester of Formula (V) in order to form a compound of Formula (VI);OH O XzW>LO(V) (VI)wherein Ri is optionally substituted alkyl; andb) hydrolysing the ester in order to form the compound of Formula (I).

2. The method according to claim 1, wherein step a) comprises reacting the compound of Formula (V) with a compound of Formula (VII) to form a compound of Formula (Via).OH O(V) wherein X is halo.

3. The method according to claim 1, wherein step a) comprises oxidising the compound of Formula (V) into a compound of Formula (XIV), reducing the compound of Formula (XIV) in the presence of an amine to form a compound of Formula (VIII), and reacting the compound of Formula (VIII) with a compound of Formula (VII) to form a compound of Formula (VIb):Reducing agentQH Q Oxidation o O NH2- 2H'N'RzO w »-Ri“ >? W5O-R’—(v)(XIV)(VIII)wherein R2 is independently selected from H and optionally substituted alkyl; and X is halo.

4. The method according to claim 1, wherein step a) comprises halogenating followed by azidating the compound of Formula (V) to afford compound of Formula (IX), and subjecting the compound of Formula (IX) to a click reaction to form compound of Formula (Vic):

5. The method according to claim 1, wherein step a) comprises reacting a compound of Formula (V) with a compound of Formula (XI) to form a compound of Formula (Vid):(Vid) wherein X is halo.

6. The method according to claim 1, wherein step a) comprises reacting a compound of Formula (V) with a compound of Formula (XII) to form a compound of Formula (Vie):OH O(V) (XII) (Vie) wherein X is halo.

7. The method according to claim 1, wherein step a) comprises reacting a compound of Formula (V) with a compound of Formula (XIII) to form a compound of Formula (Vlf):OH O(Vlf) wherein X is halo.

8. The method according to any one of claims 1 to 7, further comprising a step before step a) of reacting a compound of Formula (II) with a compound of Formula (III) in order to form a compound of Formula (IV);(IV) wherein X is halo; andprotecting the compound of Formula (IV) as a compound of Formula (V);OH(IV) (V)wherein Ri is optionally substituted alkyl.

9. The method according to any one of claims 1 to 8, wherein the compound of Formula (I) is a compound of Formula (la-f):whereini is an integer selected from 3 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

10. The method according to any one of claims 1 to 9, wherein the compound of Formula (I) is a compound of Formula (1-1):(1-1).

11. The method according to any one of claims 1 to 10, wherein L is oxyacylene; i is an integer selected from 0 to 15;m is an integer selected from 5 to 10; andn is an integer selected from 5 to 10.

12. The method according to any one of claims 1 to 11, wherein L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;i is an integer selected from 3 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

13. A compound of Formula (I), or a salt, solvate, stereoisomer, prodrug, or derivative thereof:o(I,whereinL is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;i is an integer selected from 3 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

14. The compound according to claim 13, wherein the compound of Formula (I) is an enantiomer as represented by Formula (1-1):OW ™ (I-D.

15. The compound according to claim 13 or 14, wherein i is an integer selected from 10 to 15.

16. The compound according to any one of claims 13 to 15, wherein m is an integer selected from 10 to 15.

17. The compound according to any one of claims 13 to 16, wherein n is an integer selected from 1 to 5.

18. A composition, comprising a compound of Formula (I), according to any one of claims 13 to 17, or a salt, solvate, stereoisomer, prodrug, or derivative thereof, and optionally an excipient.

19. A method of treating a disease or condition associated with lipid accumulation and / or inhibiting lipid absorption in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, derivative or combination thereof to thesubject:whereinwhen L is oxyacylene;i is 15, m is 8, and n is 7; ori is 3, m is 10, and n is 5; ori is 13, m is 10, and n is 5; ori is 15, m is 10, and n is 5; ori is 11, m is 10, and n is 5; orwhen L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;i is an integer selected from 0 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

20. A method of remodeling a gut microbiome in a digestive system of a subject in need thereof, comprising administering a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, derivative or combination thereof to the subject:whereinwhen L is oxyacylene;i is 15, m is 8, and n is 7; ori is 3, m is 10, and n is 5; ori is 13, m is 10, and n is 5; ori is 15, m is 10, and n is 5; ori is 11, m is 10, and n is 5; orwhen L is a linker selected from oxylene, aminoacylene, oxysulfonylene, oxyacyloxyene, heterocyclylene, and heteroarylene;i is an integer selected from 0 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

21. The method according to claim 19 or 20, wherein the compound of Formula (I) is an enantiomer as represented by Formula (1-1):

22. The method according to any one of claims 19 to 21, wherein the compound of Formula (I) is selected fromwhereini is 15, m is 10, and n is 5; ori is 11, m is 10, and n is 5.

23. The method according to any one of claims 19 to 22, wherein the compound of Formula (I) is selected fromOO(Ib)' n N O(IC)i is an integer selected from 3 to 20;m is an integer selected from 0 to 15; andn is an integer selected from 0 to 15.

24. The method according to any one of claims 19, 21 to 23, wherein the disease or condition associated with lipid accumulation is selected from a metabolic disease and / or an inflammatory disease.

25. The method according to any one of claims 19, 21 to 24, wherein the disease or condition associated with lipid accumulation is selected from dyslipidemia, hypertriglyceridemia, mixed dyslipidemia, obesity, insulin resistance, impaired glucose tolerance, type 2 diabetes, metabolic syndrome, visceral adiposity, fatty liver disease including metabolic dysfunction-associated steatotic liver disease (MASLD), metabolicdysfunction-associated steatohepatitis (MASH), alcohol-associated fatty liver disease, steatotic liver with fibrosis, liver cirrhosis secondary to steatosis, pancreatic steatosis, skeletal muscle steatosis, cardiac steatosis, atherosclerotic cardiovascular disease, hyperlipidemia-associated hypertension, imbalanced gut microbiome, increased intestinal permeability, inflammatory bowel diseases including ulcerative colitis and Crohn's disease, microbiome-associated inflammatory conditions, chronic low-grade inflammation associated with ectopic lipid deposition, and inflammatory diseases associated with dyslipidemia and metabolic dysfunction.

26. The method according to any one of claims 19, 21 to 25, wherein the disease or condition associated with lipid accumulation is a multifactorial disease.

27. The method according to any one of claims 19 to 26, wherein the method is characterised by a tolerance to insulin and / or a tolerance to glucose absorption.

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