Lipid prodrugs of bromoerguramide and uses thereof

Abstract

This article discloses a lipid prodrug of bromo-lysergic acid diethylamide (2-bromo-LSD) (2-bromo-lysergic acid diethylamide), which achieves improved pharmacokinetic properties and allows for easier long-term administration to patients. The disclosed lipid prodrug is indicated for the treatment of cluster headaches and mood disorders in human subjects.

Description

[0001] Cross-reference of related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 589,107, filed October 10, 2023, and U.S. Provisional Application No. 63 / 631,812, filed April 9, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure generally relates to a lymphotropic lipid prodrug of bromolysergide (2-bromo-lyserate diethylamide) with improved pharmacokinetic properties. Background Technology

[0004] There are few effective treatment options available for pain associated with cluster headaches. Cluster headaches cause severe, unilateral temporal or periorbital pain lasting from 15 to 180 minutes, accompanied by autonomic symptoms of the nose, eyes, and face (Weaver-Agostoni, Am Fam Physician., 2013). Serotonin receptor agonists ( For example Sumatriptan, methysergide, dihydroergotamine, and psilocybin have been shown to be effective in cluster headaches, but hallucinations, fibrotic side effects, and / or short duration of action have prevented their widespread use.

[0005] Bromo-lysergic acid diethylamine (2-bromo-LSD) is an ergot alkaloid derivative, reportedly due to its inhibition of 5-HT. 2A Partial agonism of receptors without causing hallucinations (Lewis M) et al. (Cell Reports, 2023). However, when administered orally, bromolybdate diethylamine (2-bromo-LSD) has variable pharmacokinetic properties, which limits its potential therapeutic use. Summary of the Invention

[0006] This article describes a lymphotropic lipid prodrug of bromo-lysergic acid diethylamide (2-bromo-LSD) with improved properties, such as pharmacokinetic properties, which allows for the treatment of patients with cluster headaches, migraines, anxiety and / or mood disorders.

[0007] In one aspect, this article discloses a lipid prodrug of bromolybdate diethylamine (2-bromo-LSD) represented by formula (I):

[0008] Formula (I)

[0009] Or its pharmaceutically acceptable salt, wherein:

[0010] R 1 and R 2 Each is independently hydrogen or -C(O)R 3 ;

[0011] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 2-37 hydrocarbon chain;

[0012] X is -O-;

[0013] R 4 and R 5 Each of the C atoms is independently hydrogen or optionally substituted with one, two, three, four, five, or six deuterium or halogen atoms. 1-4 aliphatic group, or R 4 and R 5 Together with the carbons they are attached to, they form C3-C8 cycloalkyl groups;

[0014] M is absent or is a self-immolative group.

[0015] n is 0-18; and

[0016] m and q are each independently 0-6.

[0017] In some implementation schemes, R 1 and R 2 Each is independently -C(O)R 3 In some implementations, each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a C7 hydrocarbon chain. In some embodiments, X is O. In some embodiments, m and q are each independently 0 or 1. In some embodiments, n is 0-6. In some embodiments, R... 4 and R 5 Each is either hydrogen or methyl.

[0018] In some implementations, M is selected from M. , , , , , , , , , , , , , and .

[0019] In one respect, this article discloses a compound of formula II:

[0020] (Formula II)

[0021] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0022] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0023] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0024] In some implementations, the compound is I-19:

[0025] (I-19).

[0026] In one respect, this article discloses a compound of formula III:

[0027] (Formula III)

[0028] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0029] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0030] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0031] In some implementations, the compound is I-16:

[0032]

[0033] (I-16).

[0034] In one respect, this article discloses a compound of formula IV:

[0035] (Form IV)

[0036] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0037] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0038] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0039] In some implementations, the compound is I-9:

[0040] (I-9).

[0041] In one respect, this paper discloses a compound of formula V:

[0042] (Form V)

[0043] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0044] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0045] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0046] In some implementations, the compound is I-18:

[0047] (I-18).

[0048] In one respect, this article discloses a compound of formula VI:

[0049] (Form VI)

[0050] Where R 1 and R 2 Each is independently -C(O)R 3 ;

[0051] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 hydrocarbon chain;

[0052] R 4 and R 5 Each of the C atoms is independently hydrogen or optionally substituted with one, two, three, four, five, or six deuterium or halogen atoms. 1-4 aliphatic group, or R 4 and R 5 Together with the carbons they are attached to, they form C3-C8 cycloalkyl groups;

[0053] p is 1 or 2; and

[0054] -M- is , , , , , or .

[0055] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain. In some implementations, -M- is... In some implementations, -M- is... .

[0056] In some implementation schemes, R 4 and R 5 Together with the carbon atoms they are attached to, they form C3 cycloalkyl groups. In some embodiments, R 4 It is methyl, and R 5 It is hydrogen. In some implementations, R 4 and R 5 Each is a methyl group. In some embodiments, p is 1. In some embodiments, p is 2.

[0057] In another respect, this disclosure relates to pharmaceutically acceptable compositions comprising compounds of any one of formulas (I), (II), (III), (IV), (V) or (VI) and pharmaceutically acceptable carriers, adjuvants or mediators.

[0058] In some implementations, the pharmaceutically acceptable composition is administered orally to a subject in need.

[0059] On the other hand, this disclosure relates to a method of treating cluster headaches, migraines, anxiety disorders, or mood disorders in patients requiring treatment, comprising administering to the patient a compound of any one of formulas (I), (II), (III), (IV), (V), or (VI), or a pharmaceutically acceptable composition comprising a compound of any one of formulas (I), (II), (III), (IV), (V), or (VI). In some embodiments, the mood disorder is depression, bipolar disorder, premenstrual syndrome irritability disorder (PMDD), intermittent explosive disorder (IED), or substance-induced mood disorder.

[0060] In some implementations, the method further includes the step of administering an additional therapeutic agent. Attached Figure Description

[0061] Figure 1 This is a diagram illustrating the metabolic pathways of bromo-2-bromo-LSD in human liver microsomes (HLM) and rat and human hepatocytes (R and H, respectively). Gluc = glucuronide, and GSH = glutathione.

[0062] Figures 2A to 2D This demonstrates the effectiveness of an aqueous cyclodextrin formulation targeting 2-bromo-LSD (2-BL) in non-human primates. Figure 2A ), self-emulsifying drug delivery system (SEDDS) lipid formulation of 2-bromo-LSD ( Figure 2B ) and SEDDS lipid prodrugs of Formula I ( Figures 2C to 2D ) The variability of observed pharmacokinetic characteristics.

[0063] Figure 3 The shift in Tmax of the SEDDS lipid prodrug of Formula I compared to 2-bromo-LSD is shown.

[0064] Figure 4 The sex difference in AUC observed in the SEDDS lipid formulation for 2-bromo-LSD was shown compared to that of the SEDDS lipid prodrug of Formula I. Detailed Implementation

[0065] This article discloses a lymphotropic lipid prodrug of bromo-lysergic acid diethylamide (2-bromo-LSD) with improved pharmacokinetic and / or tolerability properties. These improved pharmacokinetic and / or tolerability properties may partially allow for long-term administration of bromo-lysergic acid diethylamide (2-bromo-LSD) while reducing central nervous system (CNS) effects.

[0066] Lymphatic system targeted prodrug

[0067] The lipid prodrug and its composition disclosed herein can be used to promote the transport of bromo-lysergic acid diethylamide (2-bromo-LSD) into the lymphatic system and subsequently the parent drug ( Right now (therapeutic agents) For example Release of bromo-lysergic acid diethylamine (2-bromo-LSD).

[0068] In one aspect, this disclosure provides a lipid prodrug of bromo-2-bromo-LSD. Without being bound by any particular theory, it is anticipated that a prodrug of bromo-2-bromo-LSD that avoids first-pass metabolism will have improved pharmacokinetic properties and / or tolerability compared to bromo-2-bromo-LSD alone. The lipid prodrug of this disclosure is transported to the lymphatic system, thereby avoiding first-pass metabolism in the liver. In bypassing first-pass metabolism, the lipid prodrug maintains high oral bioavailability of bromo-2-bromo-LSD for oral administration to a subject. Lymphatic transport also involves processing via the natural biochemical pathway of fat absorption in the intestine. Transport via these pathways is typically a longer process than standard small molecule absorption, resulting in maximum concentrations of dietary lipids absorbed within 4–5 hours after ingestion. Therefore, in some embodiments, the lipid prodrug disclosed herein can be used to treat diseases or conditions in which elevated levels of bromo-2-bromo-LSD are beneficial.

[0069] definition

[0070] While the terminology used herein is considered to be well understood by one of ordinary skill in the art, definitions are set forth in order to facilitate explanation of the subject matter currently disclosed.

[0071] As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of a disease or condition or one or more symptoms thereof as described herein, or inhibiting its progression. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered before the onset of symptoms (…). For exampleTreatment may be administered to susceptible individuals based on their symptom history and / or genetic or other predisposing factors. Treatment may also continue after symptoms have subsided, for example, to prevent or delay recurrence. In some implementations, the disease or condition is cluster headache, migraine, anxiety disorder, or mood disorder.

[0072] As used herein, the term "optionally substituted" means that a group may or may not be further substituted by one or more groups. Suitable groups for optional substitution include, but are not limited to, alkyl, alkenyl, alkynyl, and hydroxyl groups. Other suitable groups for optional substitution are described in WO2016 / 023082, WO2017 / 041139, and WO2019 / 046491, the entire contents of which are incorporated herein by reference.

[0073] As used herein, the term "aliphatic" or "aliphatic group" refers to a straight chain that is fully saturated or contains one or more unsaturated units. Right now A hydrocarbon chain that is unbranched or branched, substituted or unsubstituted. Unless otherwise stated, the aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-4 aliphatic carbon atoms. In still other embodiments, the aliphatic group contains 1-3 aliphatic carbon atoms, and in yet still other embodiments, the aliphatic group contains 1-2 aliphatic carbon atoms. Examples of aliphatic groups include, but are not limited to, methyl, ethyl, and... just propyl, different propyl, just Butyl, Uncle Butyl, Zhong Butyl and different Butyl.

[0074] The term "halogen" refers to F, Cl, Br, or I.

[0075] As used herein, the term "self-detaching group" refers to a divalent chemical moiety comprising a easily cleavable covalent bond as one of its divalent bonds and a stable covalent bond with the therapeutic agent as its other divalent bond, wherein the bond with the therapeutic agent becomes unstable upon the breaking of the easily cleavable bond. Examples of self-detaching groups include, but are not limited to, disulfide self-detaching groups, hydrazone self-detaching groups, acetal self-detaching groups, carboxyl acetal self-detaching groups, carboxyl (methyl acetal) self-detaching groups, and p-hydroxybenzyl self-detaching groups. right Hydroxyphenylmethyl carbonyl self-removing groups, inverted ester self-removing groups, and trimethylolpropionate or 2-hydroxyphenylcarbamate (2-HPC) self-removing groups. Many other suitable self-removing groups are known in the art, such as, for example, CABlencowe... et al. , Polym. Chem. 2011, 2, 773-790 and F. Kratz et al., ChemMedChem. 2008,3(1), 20-53; Huvelle, S. et al. , Org. Biomol. Chem. 2017, 15 (16), 3435–3443; and Alouane, A. et al. , Angew. Chem. Int. Ed. 2015, 54 (26), 7492–7509; and Levine, MN et al. , Chem. Sci. VL - IS - 3 As described in (8), 2412-2420; each of them is hereby incorporated in its entirety by reference.

[0076] Lipid prodrug of bromo-lysergic acid diethylamine (2-bromo-LSD)

[0077] The development of prodrugs is typically aimed at improving the pharmacokinetic (PK) and pharmacodynamic (PD) properties of therapeutic agents, including potential side effects associated with a given PK characteristic. While many types of prodrugs exist in the art, lipid prodrugs are particularly effective in addressing oral bioavailability issues. Lipid-mimicking compounds are expected to behave similarly to natural triglycerides, enabling them to be transported via the lymphatic system before reaching systemic circulation, thus effectively circumventing first-pass metabolism. Lipid prodrugs are further discussed in WO2016 / 023082 and WO2017 / 041139.

[0078] This article discloses a lipid prodrug of bromo-lysergic acid diethylamine (2-bromo-LSD) that avoids first-pass metabolism and results in systemic exposure to 2-bromo-LSD.

[0079] The following structure of bromo-lysergic acid diethylamine (2-bromo-LSD) was first synthesized in the 1950s and found to be effective for headaches (including migraines) (Sicuteri, F., Triangle 1963, 6 (116-125), but with observed central nervous system (CNS) side effects. The structure-related serotonin receptor agonist mesimergot has been commercialized as an effective treatment for migraines and cluster headaches. However, it has been found that due to the strong 5-HT of its main metabolite... 2B Due to its receptor agonist effect, long-term use of mesimergot can lead to retroperitoneal and pulmonary fibrosis; therefore, mesimergot (Koehler, P.) should be discontinued. et al. Cephalalgia2008 28 , 1126-1135).

[0080]

[0081] Bromolyzed lysergic acid diethylamine (2-bromo-LSD)

[0082] Classic psychedelic drugs (such as psilocybin) have been shown to be effective in cluster headaches, but these molecules are also 5-HT. 2B Receptor agonists, thus posing risks of long-term use, such as in the treatment of chronic cluster headaches (Schindler, EAD). et al. , Headache 2022, 62 , 1383-1394; Madsen, MK et al. , medRxiv2022.07.10.22277414;Tagen, M. et al. Psychopharmacol 2023 37 (876-890). In contrast, bromolyzedioyldiethylamine (2-bromo-LSD) is a 5-HT... 2B An antagonist, and its long-term use is not expected to cause fibrosis. Another benefit of bromo-lysergic acid diethylamide (2-bromo-LSD) compared to classic psychedelics is its serotonin activity against 5-HT. 2A The partial agonist effect of the receptor does not trigger hallucinations as strongly as the agonist effect (Lewis, V). et al. Cell Reports 2023 42 (112203). The patient has been given up to 22 mg of bromolybdate diethylamine (2-bromo-LSD) and has not reported hallucinations (Schneckloth, R). et al. Circulation 1957, 16 (523-532). However, the pharmacokinetic (PK) properties of bromolybdate diethylamine (2-bromo-LSD) may play a role in the CNS side effects identified in the 2010 cluster headache trial, which could also potentially lead to post-treatment driving impairment (Karst, M). et al. Cephalalgia 2010 30 (1140-1144). Acute CNS side effects during the trial (Karst, M). et al. (Cephalalgia 2010, 30, 1140-1144) may be due to the maximum serum concentration of the drug (C max This is caused by [the drug / method]. Furthermore, preclinical studies in mice using bromo-lysergic acid diethylamide (2-bromo-LSD) showed sex differences in PK characteristics, variable exposure and half-life, and nonlinear PK properties (Lewis, V). et al. Cell Reports 2023 42 (112203). The structurally related compound dihydroergotamine (DHE) has shown inconsistent efficacy and adverse events due to pharmacokinetic variability between formulations (Silberstein SD). et al. , "Then and Now: A Narrative Review" Headache 60:45-57 (2020)). The PK variability observed in certain headache medications (such as amitriptyline and topiramate) requires titration to ensure safety and tolerability (Tfelt-Hansen P). et al. , Pharmacokinetic Variability of DrugsUsed for Prophylactic Treatment of Migraine. CNS Drugs 31:389-403 (2017)). Additionally, preclinical studies of 2-bromo-LSD have shown sex differences in PK characteristics, which could lead to sex-specific adverse events in clinical trials (Lewis V). et al. , “A non-hallucinogenic LSD analog with therapeutic potential for mood disorders” Cell Reports 42, 112-203 (2023) and Zucker I and Prendergast BJ “Sex Differences in Pharmacokinetics Predict Adverse DrugReactions in Women” Biology of Sex Differences 11:32 (2020)). The PK variability and potential association with CNS side effects are limitations of bromo-2-bromo-LSD that can be addressed via a lipid prodrug of bromo-2-bromo-LSD as described herein, which can improve PK characteristics, reduce dosage, and potentially reduce CNS side effects.

[0083] In particular, the lipid prodrug of 2-bromo-LSD disclosed herein results in reduced variability in pharmacokinetic characteristics, such as... Figures 2A to 2D As shown in the image. Figure 2A and Figure 2B 2-bromo-LSD was demonstrated in aqueous formulations ( Figure 2A ) and SEDDS lipid formulations ( Figure 2B The variability of PK in ). In contrast, Figure 2C and Figure 2D The reduced PK variability provided by the prodrug of this disclosure is demonstrated.

[0084] Furthermore, the lipid prodrug of 2-bromo-LSD caused an observed Tmax shift from approximately 2 hours with the SEDDS formulation of 2-bromo-LSD (black circle) to approximately 4 hours with the prodrug of this disclosure (triangle), as... Figure 3 As shown in the image.

[0085] In addition, lipid prodrugs for 2-bromo-LSD minimized the sex differences in AUC levels observed in SEDDS formulations targeting 2-bromo-LSD. Figure 4 The reduction in sex differences observed with the lipid prodrug of this disclosure was shown compared with the SEDDS formulation of 2-bromo-LSD.

[0086] As used herein, the terms “bromo-lysergic acid diethylamine”, “2-bromo-LSD”, and “2BL” are used interchangeably.

[0087] Therefore, in one respect, this paper discloses compounds of formula I:

[0088] Formula (I)

[0089] Or its pharmaceutically acceptable salt, wherein:

[0090] R 1 and R 2 Each is independently hydrogen or -C(O)R 3 ;

[0091] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 2-37 hydrocarbon chain;

[0092] X is -O-;

[0093] R 4 and R 5 Each of the C atoms is independently hydrogen or optionally substituted with one, two, three, four, five, or six deuterium or halogen atoms. 1-4 aliphatic group, or R 4 and R 5 Together with the carbons they are attached to, they form C3-C8 cycloalkyl groups;

[0094] M is absent or is a self-exfoliating group;

[0095] n is 0-18; and

[0096] m and q are each independently 0-6.

[0097] In some implementation schemes, R1 and R 2 Each is independently -C(O)R 3 .

[0098] In some implementations, each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is C 17 Hydrocarbon chain. In some implementations, each R 3 It is a C7 hydrocarbon chain.

[0099] In some implementations, X is O.

[0100] In some implementations, m and q are each independently 0 or 1.

[0101] In some implementations, n is 0-6. In other implementations, n is 0.

[0102] In some implementations, the sum of n+m+q is 0, 1, 2, 3, 4, 5, 6, 7, 8, or a range between and including any two of these values. In some implementations, the sum of n+m+q is 1. In some implementations, n is 0, m is 0, and q is 1.

[0103] In some implementation schemes, R 4 and R 5 Each is either hydrogen or methyl.

[0104] In some implementations, M is absent.

[0105] In some implementations, M is selected from , , , , , , , , , , , , , and .

[0106] In some implementations, M is , , , , , or .

[0107] In some embodiments, the compound of formula I (CPD) is selected from Table 1:

[0108] Table 1: Exemplary Compounds

[0109]

[0110]

[0111]

[0112]

[0113]

[0114] Or its pharmaceutically acceptable salt.

[0115] The compounds of formula I above are described as being in R 1 and R 2 The position contains oleic acid or octanoic acid residues. It should be understood that many fatty acid residues are applicable to the compounds disclosed herein. Therefore, compounds of Formula I can be prepared with any suitable fatty acid residues and are considered to be within the scope of this disclosure. Examples of fatty acids include, but are not limited to, saturated straight-chain fatty acids, saturated branched-chain fatty acids, unsaturated fatty acids, hydroxy fatty acids, and polycarboxylic acids. Fatty acids and their residues applicable to the compounds disclosed herein are further described in WO2016 / 023082, WO2017 / 041139, and WO2019 / 046491.

[0116] In one respect, this article discloses a compound of formula II:

[0117] (Formula II)

[0118] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0119] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0120] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0121] In one respect, the compound of formula II is:

[0122] (I-19).

[0123] In one respect, this article discloses a compound of formula III:

[0124] (Formula III)

[0125] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0126] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0127] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0128] In one respect, the compound of formula III is:

[0129]

[0130] (I-16).

[0131] In one respect, this article discloses a compound of formula IV:

[0132] (Form IV)

[0133] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0134] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0135] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0136] In one respect, the compound of formula IV is:

[0137] (I-9).

[0138] In one respect, this paper discloses a compound of formula V:

[0139] (Form V)

[0140] Where R 1 and R 2 Each is independently -C(O)R 3 ;and

[0141] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

[0142] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain.

[0143] In one respect, the compound of formula V is:

[0144] (I-18).

[0145] In one respect, this article discloses a compound of formula VI:

[0146] (Form VI)

[0147] Where R 1 and R 2 Each is independently -C(O)R 3 ;

[0148] Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 hydrocarbon chain;

[0149] R 4 and R 5 Each of the C atoms is independently hydrogen or optionally substituted with one, two, three, four, five, or six deuterium or halogen atoms. 1-4 aliphatic group, or R 4 and R 5 Together with the carbons they are attached to, they form C3-C8 cycloalkyl groups;

[0150] p is 1 or 2; and

[0151] -M- is , , , , , or .

[0152] In some implementations, each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain. In some implementations, each R 3 It is a saturated C7 hydrocarbon chain. In some implementations, -M- is... In some implementations, -M- is... .

[0153] In some implementation schemes, R 4 and R 5 Together with the carbon atoms they are attached to, they form C3 cycloalkyl groups. In some embodiments, R 4 It is methyl, and R 5 It is hydrogen. In some implementations, R 4 and R 5 Each is a methyl group. In some embodiments, p is 1. In some embodiments, p is 2.

[0154] As used herein, the term "pharmaceutically acceptable salt" refers to salts that, within reasonable medical judgment, are suitable for contact with tissues of humans and lower animals without undue toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail by SM Berge et al. in J. Pharmaceutical Sciences, 1977, 66, 1-19 (which is incorporated herein by reference).

[0155] Unless otherwise stated, the structures described herein are also intended to include all heterogeneities of that structure. For example Enantiomers, diastereomers, and geometric (or conformations) are possible forms; for example, R and S configurations, Z and E double bond isomers, and Z and E conformational isomers for each asymmetric center. Therefore, single stereochemical isomers of the compounds of the present invention, as well as enantiomers, diastereomers, and mixtures of geometric (or conformations), are within the scope of the present invention. Unless otherwise stated, all tautomers of the compounds of this disclosure are within the scope of this disclosure. Unless otherwise defined, the absolute stereochemistry of the bromolybdate diethylamine (2-bromo-LSD) moiety of the prodrug described herein is as depicted. Additionally, unless otherwise stated, the structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, those having hydrogen replaced by deuterium or tritium, or... 13 C or14 Compounds of the present invention with carbon-enriched carbon-substituted carbon structures are within the scope of this invention. Such compounds can be used, for example, as analytical tools, as probes in bioassays, or as therapeutic agents according to this disclosure.

[0156] Pharmaceutical Composition

[0157] According to another embodiment, this disclosure provides a pharmaceutical composition comprising the lipid prodrug of this disclosure and a pharmaceutically acceptable carrier, adjuvant, or mediator. The amount of the lipid prodrug in the composition is an amount effective in treating the relevant disease, condition, or illness of a patient in need (“effective amount”). In some embodiments, the compositions of this disclosure are formulated for oral administration to a patient.

[0158] As used in this article, the term "patient" refers to an animal, such as a mammal, like a human.

[0159] The term "pharmaceutically acceptable carrier, adjuvant, or mediator" refers to a non-toxic carrier, adjuvant, or mediator that does not impair the pharmacological activity of the drug formulation with which it is formulated. Therefore, the lipid prodrugs of this disclosure can be prepared into compositions with one or more pharmaceutically acceptable carriers, adjuvants, or mediators known in the art. To aid delivery, the lipid prodrugs of this disclosure can be formulated in lipid-based formulations.

[0160] The compositions disclosed herein can be administered orally. Lipid-based formulations for oral delivery are known in the art and may include, for example, a substantially non-aqueous medium that typically contains one or more lipid components. Lipid mediators and the resulting lipid formulations can be usefully classified according to the Lipid Formulation Classification System (LFCS) based on their shared common characteristics as described below (Pouton, CW, Eur. J. Pharm. Sci. 11 (Supplement 2), S93-S98, 2000; Pouton, CW, Eur. J. Pharm. Sci. 29 278-287, 2006).

[0161] Lipid formulations may contain lipids and / or surfactants, optionally with a co-solvent, and are generally classified into four types. Type I formulations include lipids that require digestion, such as monoglycerides, diglycerides, and triglycerides, and combinations thereof. Type II formulations are water-insoluble self-emulsifying drug delivery systems (SEDDS) that contain lipids in addition to water-insoluble surfactants. Type III formulations are SEDDS or self-microemulsifying drug delivery systems (SMEDDS) that contain lipids in addition to water-soluble surfactants and / or co-solvents. Type IV formulations primarily contain hydrophilic surfactants and co-solvents, such as PEG and propylene glycol.

[0162] The formulation may also contain materials commonly known to those skilled in the art as being included in lipid-based formulations, including antioxidants such as butylated hydroxyanisole (BHA) or butylated hydroxytoluene (BHT) and curing agents such as microporous silica, such as magnesium aluminum metasilicate (Neusilin®).

[0163] In some embodiments, the lipid prodrug may be administered orally in conjunction with an enzyme inhibitor to increase the stability of the prodrug in the gastrointestinal tract or intestinal epithelial cells. In some embodiments, the enzyme inhibitor inhibits pancreatic lipases, examples of which include, but are not limited to, Alli® (orlistat). In other embodiments, it is envisioned that the enzyme inhibitor will inhibit cellular lipases, such as monoacylglycerol lipases, examples of which include, but are not limited to, JZL184 (4-nitrophenyl-4-[bis(1,3-benzodioxane-5-yl)(hydroxy)methyl]piperidine-1-carboxylate).

[0164] Uses of lymphatic-directed lipid prodrugs

[0165] This article discloses a lymphopathic lipid prodrug of bromo-lysergic diethylamine (2-bromo-LSD) and its pharmaceutically acceptable compositions thereof.

[0166] Therefore, this disclosure provides a method for treating a patient's disease, symptom, or condition. For example Cluster headaches, migraines, anxiety disorders, or mood disorders ( For example The method involves administering the disclosed lipid prodrug to the patient, for conditions including depression (including treatment-resistant depression), bipolar disorder, premenstrual syndrome irritability disorder (PMDD), intermittent explosive disorder (IED), or substance-induced mood disorders. For example A lipid prodrug of bromo-lysergic acid diethylamine (2-bromo-LSD) or a pharmaceutically acceptable salt thereof.

[0167] Currently disclosed lipid prodrugs ( For example The lipid prodrug form of bromo-lysine diethylamine (2-bromo-LSD) can be used to stably transport the drug to the intestinal lymph and release the drug in the lymph, lymphoid tissue, tissues with high lipase activity (such as adipose tissue), liver or systemic circulation.

[0168] In some embodiments, this disclosure provides a method for improving the pharmacokinetic characteristics of bromo-lysergic acid diethylamine (2-bromo-LSD). In some embodiments, this disclosure provides a method of administering bromo-lysergic acid diethylamine (2-bromo-LSD) in a manner that improves the bioavailability of bromo-lysergic acid diethylamine (2-bromo-LSD), the method comprising administering an effective amount of the disclosed lipid prodrug form of bromo-lysergic acid diethylamine (2-bromo-LSD) to a patient in need. In some embodiments, compositions comprising the lipid prodrug form of bromo-lysergic acid diethylamine (2-bromo-LSD) described herein can be used to improve the pharmacokinetic characteristics of bromo-lysergic acid diethylamine (2-bromo-LSD) after administration to a subject.

[0169] In some embodiments, this disclosure provides a therapeutic index for improving bromolyzedioyldiethylamine (2-bromo-LSD). Right now The present disclosure provides a method for administering bromo-2-bromo-LSD (a dose range within which the therapeutic agent is effective without unacceptable adverse events). In some embodiments, the present disclosure provides a method of administering bromo-2-bromo-LSD, wherein the therapeutic index of bromo-2-bromo-LSD is improved, the method comprising administering an effective amount of the disclosed lipid prodrug form of bromo-2-bromo-LSD to a patient in need. In some embodiments, compositions comprising the lipid prodrug form of bromo-2-bromo-LSD described herein may be used to improve the therapeutic index of bromo-2-bromo-LSD after administration.

[0170] In some embodiments, this disclosure provides a method for treating, reducing, improving, or eliminating one or more symptoms in a patient associated with cluster headaches and / or migraines, the method comprising administering to the patient a prodrug or a compound of formula (I), (II), (III), (IV), (V), or (VI), or a pharmaceutically acceptable composition comprising such prodrug or a compound of formula (I), (II), (III), (IV), (V), or (VI). In some embodiments, this disclosure provides a method for treating, reducing, improving, or eliminating one or more symptoms in a patient associated with a mood disorder or anxiety disorder, the method comprising administering to the patient a compound of formula (I), (II), (III), (IV), (V), or (VI), or a pharmaceutically acceptable composition comprising such a compound of formula (I), (II), (III), (IV), (V), or (VI). In some embodiments, the mood disorder is depression, bipolar disorder, premenstrual syndrome irritability disorder (PMDD), intermittent explosive disorder (IED), or substance-induced mood disorder. In some further embodiments, the mood disorder includes depressive disorders and / or anxiety disorders. In some implementations, the depressive disorder is major depressive disorder (MDD), persistent depressive disorder (PDD), bipolar disorder, seasonal affective disorder (SAD), and / or post-traumatic stress disorder (PTSD). In some implementations, the depressive disorder is treatment-resistant depression (TRD). In some implementations, the anxiety disorder is generalized anxiety disorder, social anxiety disorder, separation anxiety disorder, or one or more phobias. In some implementations, the prodrug is selected from Table 1.

[0171] Combination therapy

[0172] The provided lipid prodrug or its pharmaceutically acceptable composition may be administered in combination with one or more other therapeutic agents and / or treatments to patients in need.

[0173] Lipid prodrugs or pharmaceutically acceptable compositions thereof may be administered alone or in combination with one or more other therapeutic compounds. Possible combination therapies may take the form of a fixed combination, or the administration of lipid prodrugs or compositions and one or more other therapeutic compounds may be interspersed or administered independently of each other, or a fixed combination and one or more other therapeutic compounds may be administered. As described above, long-term therapy is equally possible in the context of other treatment strategies as adjunctive therapy.

[0174] Such additional agents may be administered separately from the provided lipid prodrug or composition as part of a multi-dosing regimen. Alternatively, those agents may be part of a single dosage form, mixed together with the disclosed lipid prodrug in a single composition. If administered as part of a multi-dosing regimen, the two active agents may be administered simultaneously, sequentially, or at intervals between each other.

[0175] As used herein, the terms "combination," "combined," and related terms refer to the simultaneous or sequential administration of therapeutic agents according to this disclosure. For example, the disclosed lipid prodrug may be administered simultaneously or sequentially with one or more other therapeutic agents in separate unit dosage forms or together in a single unit dosage form. Therefore, this disclosure provides a single unit dosage form comprising the disclosed lipid prodrug, one or more additional therapeutic agents, and a pharmaceutically acceptable carrier, adjuvant, or mediator. In some embodiments, the additional agent is formulated in a composition separate from the lipid prodrug. In some embodiments, one or more additional agents are lipid prodrugs. For example , as described herein or prepared according to the methods described herein, lipid prodrugs.

[0176] The amount of additional therapeutic agent present in the compositions disclosed herein will not exceed the amount typically applied in compositions containing the therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the currently disclosed compositions will range from about 50% to 100% of the amount typically present in compositions containing the pharmaceutical agent as the sole active agent.

[0177] Examples of pharmaceutical agents that can be combined with the lipid prodrugs of this disclosure include, but are not limited to: therapeutic agents for cluster headaches and / or migraines, such as oxygen, triptans ( For example Sumatriptan and zolmitriptan, octreotide, local anesthetics ( For example Lidocaine, dihydroergotamine, calcium channel blockers For example Verapamil, corticosteroids (e.g., prednisone), anti-CGRP antibodies ( For example galcanezumab, lithium carbonate (LITHOBID®), antiepileptic drugs ( For example Topiramate, caffeine, ergotamine, melatonin, capsaicin, zucapsaicin, and / or sodium valproate; and medications used to treat mood disorders ( For exampleDepression (including treatment-resistant depression), bipolar disorder, premenstrual syndrome irritability disorder (PMDD), intermittent explosive disorder (IED), or substance-induced mood disorders, such as sodium valproate, carbamazepine, oxcarbazepine, lamotrigine, benzodiazepines ( example like Alprazolam, chlordiazepoxide, clobazam, clonazepam, clonazepam, clorazepate, diazepam, estazolam, flurazepam, lorazepam, midazolam, oxazepam, quazepam, remimazolam, temazepam, and triazolam), triiodothyronine (thyroid hormone), and selective serotonin reuptake inhibitors (SSRIs) For example Sertraline, fluvoxamine, fluoxetine, citalopram, escitalopram, and paroxetine, and serotonin-norepinephrine reuptake inhibitors (SNRIs) For example Venlafaxine, desvenlafaxine, duloxetine, milnacipran, levomilnacipran, and atypical antidepressants ( For example Bupropion, mirtazapine, tricyclic antidepressants (TCAs) For example Desipramine, nortriptyline, imipramine, and amitriptyline, serotonin modulators ( For example nefazodone, trazodone, vilazodone, and vortioxetine, monoamine oxidase inhibitors (MAOIs) For exampleThe treatment agents include tranylcypromine, phenelzine, selegiline, and aripiprazole. In some implementations, an additional therapeutic agent is a serotonin receptor agonist.

[0178] General methods for preparing lipid prodrugs

[0179] The lipid prodrug compounds of the present invention can generally be prepared or isolated by methods known to those skilled in the art for the synthesis and / or semi-synthesis of similar compounds, as well as by methods described in detail in the embodiments herein.

[0180] Included in the disclosed lipid prodrug ( For example Therapeutic agents (combined with glycerol ester-based prodrugs) can be commercially available or prepared by organic synthesis, semi-synthesis and similar methods known in the art.

[0181] In some implementations, protecting groups (as defined below) can be used to manipulate the therapeutic agent to prepare a conjugate with the remainder of the lipid prodrug structure, for example, to prevent unwanted side effects.

[0182] To provide a more complete understanding of the invention described herein, the following embodiments are illustrated. It should be understood that these embodiments are for illustrative purposes only and should not be construed as limiting the invention in any way.

[0183] Example

[0184] The following examples describe the synthesis of prodrugs (including intermediates) used in the methods disclosed herein. As used throughout, “eq” refers to molar equivalents. Unless otherwise stated, intermediate numbers lacking any prefix (e.g., 1, 2, 3, etc.) are specific for each scheme (or structural diagram) in which they appear, while intermediate numbers including the prefix “Int” (e.g., Int-1, Int-2, Int-3, etc.) are used to refer to the same intermediate across schemes (or structural diagrams).

[0185] As used throughout the text, 2BL refers to bromo-lysergic acid diethylamine (2-bromo-LSD):

[0186]

[0187] Example 1: Acid Intermediate

[0188] Synthesis of Int-1 (C5bMe-acid-2-TG-oleate)

[0189]

[0190] The synthesis of Int-1 is described in WO2021 / 159021 (see paragraphs

[00335] to

[00336] ).

[0191] Synthesis of Int-2 (C5bMe-acid-2-TG-octanoate)

[0192]

[0193] The synthesis of Int-2 is described in WO2021 / 159021 (see paragraphs

[00761] to

[00762] ).

[0194] Scheme 1: Synthesis of Int-3 (C5bbGMe-acid-2-TG-octanoate)

[0195]

[0196] 4-Dimethylaminopyridine (DMAP, 1.19 g, 9.71 mmol) was added to a solution of 3,3-dimethylglutaric anhydride 1 (1.38 g, 9.71 mmol) and 1,3-DG-octanoate 2 (2.23 g, 6.47 mmol, see paragraphs

[00545] to

[00546] of WO2021 / 159021) in dichloromethane (DCM, 40 mL). The mixture was stirred at room temperature (RT) for 3 days and then concentrated to give crude Int-3 as an oil. The crude product was purified by normal phase purification (Biotage Isolera, 120 g, SiliaSep filter cartridge) using 0–20% ethyl acetate / heptane eluent for 14 column volumes (CV) to give pure Int-3 as a colorless oil (951 mg, 30.2% yield).

[0197] ¹H NMR (400 MHz, CDCl3): δ 5.31 - 5.25 (m, 1H), 4.29 (dd, J = 4.2, 12.0 Hz, 2H), 4.14 (dd, J = 6.1, 12.0 Hz, 2H), 2.47 (s, 2H), 2.46 (s, 2H), 2.30 (t, J = 7.6 Hz, 4H), 1.65 - 1.54 (m, 4H), 1.35 - 1.21 (m, 16H), 1.14 (s, 6H), 0.89 - 0.84 (m, 6H); No exchangeable CO2H protons were observed. (via 7.6 Hz, 4H), 1.65 - 1.54 (m, 4H), 1.35 - 1.21 (m, 16H), 1.14 (s, 6H), 0.89 - 0.84 (m, 6H); No exchangeable CO2H protons were observed. 1 H NMR, the compound contains 20.2% (w / w%) 3,3-dimethylglutaric anhydride and 1.7% (w / w) DCM.

[0198] UPLC-MS: (XB BEH300 C4 20-95%): R t = 3.58 minutes, 97.3% (UV), 100.0% (ELSD). MS (ESIpos): m / z = 504.7 [M+NH4] + ; MS (ESIneg): m / z = 485.7 [MH] - .

[0199] Option 2: Synthesis of Int-4 (C5bcPr-acid-2-TG-oleate)

[0200]

[0201] DMAP (1.50 g, 12.3 mmol) was added to a solution of 6-oxaspiro[2.5]octane-5,7-dione 1 (1.69 g, 12.1 mmol) and 1,3-DG-oleate 2 (5.00 g, 8.05 mmol, see paragraphs

[00526] to

[00528] of WO2021 / 159021) in DCM (60 mL). The mixture was stirred at room temperature for 5 days and then concentrated to give crude Int-4. The crude product was purified by normal phase purification (Biotage Isolera, 120 g SiliaSep filter cartridge) using 40–60% ethyl acetate / heptane eluent for 15 CV cycles to give pure Int-4 (3.57 g, 58.3% yield) as a colorless oil.

[0202] 1 H NMR (400 MHz, CDCl3): δ 5.46 - 5.22 (m, 5H), 4.31 (dd, J = 4.3,11.9 Hz, 2H), 4.18 - 4.11 (m, 2H), 2.46 (s, 2H), 2.45 (s, 2H), 2.31 (t, J =7.5 Hz, 4H), 2.07 - 1.97 (m, 8H), 1.68 - 1.56 (m, 4H), 1.37 - 1.22 (m, 40H), 0.92 - 0.83 (m, 6H), 0.57 (s, 4H). Expected 80H, observed 79H, no exchangeable CO2H protons observed.

[0203] 13C NMR (100 MHz, CDCl3): δ 177.2, 173.4, 171.4, 130.2, 129.9, 69.3, 62.2, 41.1, 40.8, 34.2, 32.1, 29.91, 29.85, 29.7, 29.5, 29.31, 29.26, 29.24,27.4, 27.3, 25.0, 22.8, 14.3, 14.2, 12.4.

[0204] UPLC-MS (XB BEH300 C4 20 to 95%): R t = 6.32 minutes, 100.0% (UV), 99.9% (ELSD); MS (ESIpos): m / z = 779.1 [M+H] + , MS(ESIneg): m / z = 760.1 [MH] - .

[0205] Scheme 3: Synthesis of Int-5 (C5bcPr-acid-2-TG-octanoate)

[0206]

[0207] DMAP (1.36 g, 11.1 mmol) was added to a solution of 6-oxaspiro[2.5]octane-5,7-dione 1 (1.53 g, 11.0 mmol) and 1,3-DG-octanoate 2 (2.51 g, 7.30 mmol) in DCM (50 mL). The mixture was stirred at room temperature for 40 hours and concentrated to give crude Int-5. The crude product was purified by normal phase purification (Biotage Isolera, 120 g SiliaSep filter cartridge) using 0–40% ethyl acetate / heptane eluent for 12 CV cycles to give pure Int-5 (1.10 g, 31.1% yield) as a colorless oil.

[0208] ¹H NMR (400 MHz, CDCl3): δ 5.27 (quin, J = 5.0 Hz, 1H), 4.37 - 4.25(m, 2H), 4.14 (dd, J = 5.9, 11.9 Hz, 2H), 2.51 - 2.40 (m, 4H), 2.31 (t, J=7.5 Hz, 4H), 1.67 - 1.55 (m, 4H), 1.34 - 1.21 (m, 16H), 0.90 - 0.83 (m, 6H), 0.57 (s, 4H). Expected 44H, observed 43H. No exchangeable CO2H protons were observed.

[0209] 13 C NMR (101 MHz, CDCl3): δ 173.5, 171.5, 171.4, 69.3, 62.2, 41.0, 40.9, 40.8, 34.2, 31.8, 29.2, 29.0, 25.0, 22.7, 14.2, 12.4.

[0210] UPLC-MS: (XB BEH 300 C4 20-95%): R t = 3.35 minutes, 92.4% (UV), 100.0% (ELSD); MS (ESIpos): m / z = 502.7 [M+H] + .

[0211] Scheme 4: Synthesis of Int-6 (C6bbGMe-acid-2-TG-octanoate)

[0212]

[0213] 3-chloroperoxybenzoic acid (mCPBA, 4.64 g, 26.9 mmol) was added to a solution of 4,4-dimethylcyclohexanone 1 (2.26 g, 17.9 mmol) in DCM (50 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The resulting white solid was filtered off and washed with DCM (3 × 50 mL). The filtrate was washed with an aqueous solution of NaHCO3 (3 × 100 mL). The organic phase was dried over MgSO4, filtered, and concentrated to give 2 (2.60 g, crude substance) as a white solid. The product was used in the next step without further purification.

[0214] 1 H NMR (400 MHz, CDCl3): δ 4.21-4.19 (m, 2H), 2.60 (t, J = 6.0 Hz, 2H), 1.64 (t, J = 4.8 Hz, 2H), 1.56 (t, J = 6.0 Hz, 2H), 1.02 (s, 6H).

[0215] Sodium hydroxide (612 mg, 15.3 mmol) was added in a single addition to a solution of 5,5-dimethyloxacycloheptan-2-one 2 (1.50 g, 10.6 mmol) in water (10 mL) at room temperature. The reaction mixture was stirred at 70°C for 16 hours. The reaction mixture was cooled to room temperature and the pH was adjusted to 3 with 2N HCl. The organic matter was extracted with diethyl ether (3 × 50 mL). The combined organic matter was washed with brine (100 mL), dried over MgSO4, filtered, and concentrated to give a colorless, gel-like product 3 (1.35 g, crude matter). The product was used in the next step without further purification.

[0216] 1 H NMR (400 MHz, CDCl3): δ 6.70 – 6.30 (br s, 1H), 3.71 (t, J = 8.8Hz, 2H), 2.32 (t, J = 7.4 Hz, 2H), 1.59 (t, J = 8.2 Hz, 2H), 1.52 (t, J = 7.6 Hz, 2H), 0.94 (s, 6H); No exchangeable CO2H protons were observed.

[0217] 13 C NMR (100 MHz, CDCl3): δ 179.7, 133.7, 130.3, 129.9, 128.3, 65.0,59.6, 43.9, 43.8, 42.0, 36.8, 35.8, 32.1, 32.0, 30.2, 29.5, 27.2.

[0218] DMAP (1.54 g, 12.6 mmol), EDC-HCl (2.42 g, 12.6 mmol), and benzyl alcohol (9.11 g, 84.3 mmol, 8.72 mL) were added to a solution of 6-hydroxy-4,4-dimethyl-hexanoic acid 3 (1.35 g, 8.43 mmol) in DCM (150 mL), and the mixture was heated to 50°C for 16 hours. The reaction mixture was cooled to room temperature and 2N HCl aqueous solution (50 mL) was added. The organic matter was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, filtered, and concentrated to give a crude residue. The residue was purified by normal-phase chromatography (Biotage Isolera, 120 g SiliCycle filter cartridge; eluent 0-100% ethyl acetate / heptane, 10 CVs) to give 4 (721 mg, 33% yield) as a colorless oil.

[0219] 1 H NMR (400 MHz, CDCl3): δ 7.39 – 7.30 (m, 5H), 5.11 (s, 2H), 3.69 (t,J= 8 Hz, 2H), 2.36 – 2.32 (m, 2H), 1.64 – 1.57 (m, 2H), 1.56 – 1.46 (m, 2H), 1.30 (s, 1H), 0.90 (s, 6H).

[0220] 13 C NMR (100 MHz, CDCl3): δ 174.2, 136.1, 128.7, 128.4, 66.4, 59.7, 44.2, 37.0, 32.1, 29.7, 27.2.

[0221] A solution of 6-hydroxy-4,4-dimethyl-hexanoate benzyl ester 4 (721 mg, 2.88 mmol) in acetonitrile (2 mL) was treated with potassium persulfate (2.30 g, 3.74 mmol) and IBX (242 mg, 864 μmol). The reaction mixture was heated at 70°C and stirred for 4 hours. The reaction mixture was cooled to room temperature and 2N HCl aqueous solution (20 mL) was added. The mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over MgSO4, filtered, and concentrated. The substance was purified by normal-phase chromatography (Biotage Isolera, 25 g SiliCycle filter cartridge; eluent 0-100% ethyl acetate / heptane, 15 CVs) to give 5 (384 mg, 38% yield) as a colorless oil.

[0222] 1 ¹H NMR (400 MHz, CDCl₃): δ 7.40 – 7.30 (m, 5H), 5.11 (s, 2H), 2.40 – 2.36 (m, 2H), 2.23 (s, 2H), 1.77 – 1.72 (m, 2H), 1.03 (s, 6H); No exchangeable CO₂H protons were observed.

[0223] C_UPLC2-MS: (CSH-C18 long neutral 2-95%): R t = 2.22 minutes (75.2%), MS (ESIneg): m / z = [MH] - 263.2.

[0224] EDC-HCl (557 mg, 2.91 mmol) and DMAP (355 mg, 2.91 mmol) were added to a solution of 6-benzyloxy-3,3-dimethyl-6-oxo-hexanoic acid 5 (384 mg, 1.45 mmol) and 1,3-DG-octanoate (500 mg, 1.45 mmol) in DCM (10 mL). The reaction mixture was stirred at 30°C for 16 hours. The reaction mixture was cooled to room temperature and quenched with a saturated aqueous solution of NH4Cl (50 mL). The mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, filtered, and concentrated to give a crude residue. The residue was purified by normal-phase chromatography (Biotage Isolera, 40 g SiliCycle filter cartridge; eluent 0-40% ethyl acetate / heptane, 15 CVs) to give 6 (446 mg, 41% yield) as a colorless oil.

[0225] UPLC4-MS (XB BEH300 C4 20 to 95%): R t = 5.56 minutes, 71.6% (UV), 79.3% (ELSD); MS (ESIpos): m / z = 608.8 [M+NH4] + .

[0226] 1 H NMR (400 MHz, CDCl3): δ 7.38 - 7.30 (m, 5H), 5.29 - 5.24 (m, 1H), 5.11 (s, 2H), 4.28 (dd, J = 4.2, 11.9 Hz, 2H), 4.12 (dd, J = 6.1, 11.9 Hz,2H), 2.39 - 2.35 (m, 2H), 2.29 (t, J = 7.6 Hz, 4H), 2.22 (s, 2H), 1.73 - 1.71(m, 2H), 1.61 - 1.58 (m, 4H), 1.28 - 1.27 (m, 16H), 1.00 (s, 6H), 0.93 - 0.86(m, 6H).

[0227] Under argon atmosphere, palladium on carbon (5% w / w) (803 mg, 377 μmol) was added to a solution of 6-benzyl 3,3-dimethyladipate 1-(1,3-bis(octanoyloxy)propyl-2-yl ester) 6 (446 mg, 755 μmol) in methanol (20 mL). A hydrogen atmosphere was introduced, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the organic matter was concentrated to give a yellow oil. The crude oil was passed through a silica stopper to give Int-6 (325 mg, crude matter).

[0228] UPLC4-MS (XB BEH300 C4 20 to 95%): R t = 3.90 minutes, 55.2% (UV), 98.9% (ELSD); MS (ESIpos): m / z = 501.7 [M+H] + , 518.7 [M+NH4] + .

[0229] 1 H NMR (400 MHz, CDCl3): δ 5.30 - 5.25 (m, 1H), 4.28 (dd,J = 4.2, 11.9 Hz, 2H), 4.13 (dd, J = 6.1, 11.9 Hz, 2H), 2.38 - 2.35 (m, 2H), 2.30 (t,J = 8 Hz, 4H), 2.23 (s, 2H), 1.72 - 1.68 (m, 2H), 1.60 (br t, J = 7.2 Hz, 4H), 1.28 (br s, 16H), 1.02 (s, 6H), 0.89 - 0.86 (m, 6H); No exchangeable CO2H protons were observed.

[0230] 13 C NMR (100 MHz, CDCl3): δ 179.7, 173.6, 171.1, 69.0, 62.3, 45.7,37.1, 34.2, 33.2, 31.8, 30.4, 29.2, 29.0, 27.2, 25.0, 22.7, 14.2.

[0231] Synthesis of Int-16 (C8bMe-acid-2-TG-oleate)

[0232]

[0233] The synthesis of Int-16 is described in WO2021 / 159021 (see paragraphs

[00397] through

[00399] ).

[0234] Synthesis of Int-17 (C12b'bMe-acid-2-TG-oleate)

[0235]

[0236] The synthesis of Int-17 is described in WO2021 / 159021 (see paragraphs

[00633] through

[00636] ).

[0237] Synthesis of Int-18 (C12bMe-acid-2-TG-oleate)

[0238]

[0239] The synthesis of Int-18 is described in WO2021 / 159021 (see paragraphs

[00409] through

[00413] ).

[0240] Synthesis of Int-19 (C10b'bMe-acid-2-TG-oleate)

[0241]

[0242] The synthesis of Int-19 is described in WO2021 / 159021 (see paragraphs

[00626] to

[00632] ).

[0243] Example 2: Synthesis of bromolybdate diethylamine (2-bromo-LSD) lipid prodrug

[0244] Scheme 1: Synthesis of compound I-1 (2BL-C5bMe-2-TG-oleate)

[0245]

[0246] Int-1 (500 mg, 667 μmol) was administered at 0°C in DCM (4 mL) and N,N- A solution of oxaloyl chloride (436 mg, 3.44 mmol, 300 μL) in DCM (10 mL) was added dropwise to a solution of dimethylformamide (DMF, 2 drops), and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated to give 1 (645 mg, 78.4% yield) as an orange oil.

[0247] 1 H NMR (400 MHz, CDCl3): δ 5.36 - 5.26 (m, 5H), 4.33 (dd, J = 11.9,4.1 Hz, 2H), 4.13 (ddd, J = 12.0, 6.1, 1.1 Hz, 2H), 3.11 - 2.73 (m, 2H), 2.60- 2.48 (m, 1H), 2.44 - 2.27 (m, 6H), 2.08 - 1.96 (m, 8H), 1.65 - 1.56 (m,4H), 1.38 - 1.23 (m, 40H), 1.08 (d, J = 6.4 Hz, 3H), 0.93 - 0.85 (m, 6H).

[0248] DMAP (2.00 mg, 16.4 μmol) and triethylamine (8.71 mg, 86.1 μmol, 12.0 μL) were added to a stirred solution of 2BL (20.0 mg, 49.7 μmol) in DCM (1.5 mL). The mixture was cooled to 0°C and a solution of 1 (35.0 mg, 45.6 μmol) in DCM (0.5 mL) was added, and the mixture was stirred at room temperature for 18 hours. The mixture was quenched with a saturated aqueous solution of NH4Cl (10 mL) and diluted with ethyl acetate (10 mL). The layers were separated, and the organic layer was washed with brine (10 mL), dried over MgSO4, filtered, and concentrated under vacuum. The substance was purified by normal phase chromatography (Biotage Isolera, 4 g Silisep filter cartridge; eluent 50-100% ethyl acetate / heptane [6 CVs 50%, 20 CVs 50-100%]) to give compound I-1 (3 mg, 5.3% yield) as a yellow oil.

[0249] UPLC4-MS: (XB BEH 300 C4 20-95%): Rt = 7.20 min, 40.5% (UV), 74.3% (ELSD); MS (ESIpos): m / z = 1133.5 / 1135.4 [M+H]+; 79Br / 81Br isotope mode observed.

[0250] ¹H NMR (400 MHz, CDCl3): δ 7.99 (br d, J = 7.8 Hz, 1H), 7.39 - 7.34(m, 1H), 7.33 - 7.27 (m, 1H), 6.39 (s, 1H), 5.40 - 5.23 (m, 5H), 4.30 (ddd, J= 1.6, 4.4, 11.9 Hz, 2H), 4.21 - 4.02 (m, 2H), 3.57 - 3.36 (m, 5H), 3.33 -3.22 (m, 2H), 3.20 - 3.08 (m, 2H), 2.81 - 2.69 (m, 2H), 2.62 - 2.51 (m, 1H), 2.46 - 2.36 (m, 1H), 2.29 (t, J = 7.6 Hz, 4H), 2.08 - 1.94 (m, 8H), 1.65-1.52 (br s, 9H), 1.37 - 1.23 (m, 43H), 1.18 (t, J = 7.1 Hz, 3H), 1.13 (d, J =6.9 Hz, 3H), 0.90 - 0.85 (m, 6H).

[0251] Scheme 2: Synthesis of compound I-2 (2BL-C8bMe-2-TG-oleate)

[0252]

[0253] Oxaloyl chloride (6.74 µL, 77.2 μmol) was added to a solution of Int-16 (50.0 mg, 64.3 μmol) in a mixture of DMF (10 µL, 129 µmol) and dichloromethane (2 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give 1 (51.2 mg, quantified), a pale yellow oil. Acid chloride 1 was used directly in the next reaction.

[0254] A suspension of 2BL (12.0 mg, 29.8 µmol) and sodium hydride (2.15 mg, 89.5 µmol, 60% dispersion in mineral oil) in DMF (1 mL) was heated to 50°C and stirred for 30 min. A solution of 1 (48.3 mg, 59.7 μmol) in DMF (1 mL) was added to this solution over 5 min, and the mixture was stirred at 50°C for 18 h. The reaction mixture was cooled to room temperature, diluted with ether (10 mL), and washed with water (2 × 10 mL). The organic phase was dried over MgSO4, filtered, and concentrated. The substance was purified by normal-phase chromatography (Biotage Isolera, 4 g SiliaSep filter cartridge; eluent 0-60% ethyl acetate / dichloromethane, 50 CVs) to give compound I-2 (17.0 mg, 48% yield) as a deep yellow oil.

[0255] UPLC-MS (XB BEH300 C4 20 to 95%): Rt = 7.30 min, 79.0% (UV), 99.0% (ELSD); MS (ESIpos): m / z = 1175.3, 1177.3 [M+H]+. 79Br / 81Br isotope pair observed.

[0256] 1H NMR (400 MHz, CDCl3) δ = 7.96 (d, J = 8.2 Hz, 1H), 7.36 (d, J = 7.3Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 6.37 (s, 1H), 5.38 - 5.26 (m, 5H), 4.29(dd, J = 4.2, 11.9 Hz, 2H), 4.14 (dd, J = 6.0, 11.9 Hz, 2H), 4.04 - 3.83 (m,1H), 3.49 - 3.41 (m, 4H), 3.39 – 3.05 (m, 4H), 2.96 – 2.85 (br s, 1H), 2.70 -2.43 (m, 3H), 2.37 - 2.28 (m, 5H), 2.14 (dd, J = 8.3, 14.7Hz, 1H), 2.07 -1.91 (m, 9H), 1.85 – 1.79 (m, 2H), 1.65 - 1.55 (m, 4H), 1.52 – 1.37 (m, 2H),1.36 - 1.20 (m, 47H), 1.17 (t, J = 7.1 Hz, 3H), 0.95 (d, J = 6.6 Hz, 3H),0.87 (t, J = 6.8 Hz, 6H).

[0257] 13 C NMR (100 MHz, CDCl3): δ 173.4, 172.8, 172.3, 135.6, 130.2, 129.9,126.4, 121.6, 117.6, 115.5, 69.0, 62.3, 62.0, 55.8, 43.8, 42.3, 41.7, 40.5,39.1, 36.6, 34.2, 32.0, 30.3, 29.91, 29.85, 29.80, 29.7, 29.50, 29.46, 29.31,29.26, 29.23, 27.4, 27.3, 26.6, 25.2, 25.0, 22.8, 19.6, 15.1, 14.3, 13.3。

[0258] Synthesis of compound I-3 (2BL-C12b'bMe-2-TG-oleate)

[0259]

[0260] Similar to compound I-2 as described above, compound I-3 was prepared from the intermediate C12b'bMe-acid-2-TG-oleate (Int-17) as described in Example 1.

[0261] Scheme 3: Synthesis of compound I-4 (2BL-ASI-C5bMe-2-TG-oleate)

[0262]

[0263] Water (0.2 M relative to KHSO4) containing tetra-n-butylammonium bisulfate (0.4 equivalents) and potassium bisulfate (4 equivalents) was added to a solution of Int-1 (1 equivalent) in DCM (0.05 M), and the reaction mixture was stirred at room temperature for about 30 minutes. Chloromethyl chlorosulfonate (1.2 equivalents) was added dropwise, and the reaction mixture was stirred vigorously at room temperature for about 16 hours. The reaction mixture was diluted with DCM, washed with water and brine, dried over MgSO4, and concentrated under reduced pressure to provide the crude product. Purification by silica gel chromatography with a suitable solvent mixture yielded 1.

[0264] At 0 °C, a solution of 2BL (1 equivalent) in THF (0.1 M) was added to a suspension of NaH (60% w / w dispersion in mineral oil, 1 equivalent) in THF (0.1 M), and the reaction mixture was stirred at 0 °C for about 30 minutes. A solution of 1 (1.2 equivalent) in THF (0.1 M) was slowly added to the stirred reaction mixture. The mixture was then stirred at 0 °C for 30 minutes, warmed to room temperature, and further stirred at room temperature for 4–24 hours. The reaction mixture was quenched by slow addition of a saturated aqueous NH4Cl solution, diluted with water, and extracted with ethyl acetate. The combined organic extracts were washed with water and brine, dried over MgSO4, and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography with a suitable solvent mixture gave 2BL-ASI-C5bMe-2-TG-oleate, compound I-4.

[0265] Synthesis of Compound I-5 (2BL-ASI-C8bMe-2-TG-oleate)

[0266]

[0267] Similar to compound I-4 as described above, compound I-5 was prepared from the intermediate C8bMe-acid-2-TG-oleate (Int-16) as described in Example 1.

[0268] Synthesis of compound I-6 (2BL-ASI-C12bMe-2-TG-oleate)

[0269]

[0270] Similar to compound I-4 as described above, compound I-6 was prepared from the intermediate C12bMe-acid-2-TG-oleate (Int-18) as described in Example 1.

[0271] Scheme 4: Synthesis of compound I-7 (2BL-CASI-C5bMe-2-TG-octanoate)

[0272]

[0273] Thiocarbonate O -(chloromethyl ester) S Preparation of ethyl ester reagent 1. Chloromethyl chloroformate (1.53 g, 11.9 mmol, 1.06 mL) was added dropwise to a solution of ethanethiol (739 mg, 11.9 mmol, 0.880 mL) and Et3N (1.32 g, 13.1 mmol, 1.82 mL) in diethyl ether (30 mL) at 0°C. A precipitate formed, and additional diethyl ether (30 mL) was added and the mixture was vigorously stirred. The mixture was warmed to room temperature and stirred for 66 hours. The mixture was filtered through diatomaceous earth and washed with diethyl ether (100 mL). The filtrate was concentrated to give a colorless liquid reagent (1.72 g, crude substance). The reagent was used in the next step without any further purification.

[0274] 1 H NMR (400 MHz, CDCl3): δ 5.77 (s, 2H), 2.97 - 2.89 (m, 2H), 1.34 (t,J = 7.5 Hz, 3H).

[0275] A solution of Int-2 (669 mg, 1.42 mmol) and potassium carbonate (978 mg, 7.08 mmol) in DMF (10 mL) was stirred at room temperature for 30 minutes. Thiocarbonate was then added... O -(chloromethyl ester) S A solution of ethyl ester 1 (300 mg, 1.94 mmol) in DMF (3 mL) was added to the stirred mixture and heated to 65°C for 3 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 30 mL). The organic fractions were combined, washed with water (90 mL) and brine (90 mL), dried (MgSO4), filtered, and concentrated. The substance was purified by normal phase chromatography (Biotage Isolera, 25 g SiliaSep filter cartridge; eluent 0–25% ethyl acetate / heptane, via 0% 3 CV, 0–25% 13 CV, 25% 2 CV) to give 2 (458 mg, 54% yield) as a colorless oil.

[0276] UPLC4-MS (XB BEH300 C4 20 to 95%): Rt = 5.08 min, 96.8% (UV), 98.5% (ELSD); MS (ESIpos): m / z = 609.1 [M+NH4]+.

[0277] 1 H NMR (400 MHz, CDCl3): δ 5.81 (s, 2H), 5.38 - 5.20 (m, 1H), 4.38 -4.25 (m, 2H), 4.13 (dd, J = 11.9, 6.1 Hz, 2H), 2.89 (q, J = 7.5 Hz, 2H), 2.53- 2.38 (m, 3H), 2.38 - 2.23 (m, 6H), 1.68 - 1.53 (m, 4H), 1.38 - 1.22 (m,19H), 1.07 - 1.00 (m, 3H), 0.93 - 0.84 (m, 6H).

[0278] Sulfonyl chloride (1 M, 4.10 mmol, 4.10 mL) was added dropwise to a stirred solution of 2 (237 mg, 401 μmol) in DCM (9 mL), and the mixture was heated to 40°C for 3 hours. The mixture was cooled to room temperature and then concentrated to give 3 (244 mg, crude substance) as a yellow oil. The product was used in the next step without any further purification.

[0279] 1 H NMR (400 MHz, CDCl3): δ 5.82 (s, 2H), 5.31 - 5.25 (m, 1H), 4.31 (ddd, J = 11.9, 4.1, 0.9 Hz, 2H), 4.14 (dd, J = 11.9, 6.0 Hz, 2H), 2.59 -2.26 (m, 9H), 1.69 - 1.56 (m, 4H), 1.37 - 1.22 (m, 16H), 1.13 - 1.03 (m, 3H), 0.93 - 0.83 (m, 6H).

[0280] A suspension of 2BL (83.0 mg, 206 μmol), tetrabutylammonium iodide (TBAI, 40.0 mg, 108 μmol), and NaH (17.0 mg, 425 μmol, 60% w / w dispersion in mineral oil) in tetrahydrofuran (5 mL) was stirred at 0°C for 10 min. Tetrahydrofuran (2 mL) containing 3 (234 mg, 414 μmol) was added, and the mixture was warmed to room temperature and stirred for 18 h. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined fractions were washed with brine (60 mL), dried (MgSO4), filtered, and concentrated. Normal-phase chromatography using a Biotage Isolera filter with a 4 g SiliaSep cartridge and an eluent of 50–100% ethyl acetate (+0.5% Et3N) / heptane (+0.5% Et3N), followed by two CVs at 50%, 20 CVs at 50–100%, and five CVs at 100%, yielded a yellow gel-like compound, 2BL-CASI-C5bMe-2-TG-octanoate, compound I-7 (62.0 mg, 32% yield).

[0281] UPLC4-MS (XB BEH300 C4 20 to 95%): Rt = 5.23 min, 93.5% (UV), 98.1% (ELSD); MS (ESIpos): m / z = 930.7, 932.7 [M+H]+ - 79Br / 81Br isotopic mode.

[0282] 1H NMR (400 MHz, CDCl3): δ 7.76 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 7.6Hz, 1H), 7.31 - 7.24 (m, 1H), 6.36 (s, 1H), 6.08 (s, 2H), 5.34 - 5.18 (m,1H), 4.32 - 4.24 (m, 2H), 4.11 (dd, J = 11.8, 5.9 Hz, 2H), 3.95 - 3.77 (m,1H), 3.52 - 3.32 (m, 5H), 3.24 - 3.10 (m, 1H), 3.04 (br dd, J = 11.1, 4.7 Hz,1H), 2.90 - 2.77 (m, 1H), 2.60 (s, 3H), 2.57 - 2.33 (m, 5H), 2.29 (s, 5H), 1.65 - 1.54 (m, 4H), 1.33 - 1.22 (m, 19H), 1.17 (t, J = 7.1 Hz, 3H), 1.03 (d,J = 6.4 Hz, 3H), 0.90 - 0.82 (m, 6H).

[0283] 13 C NMR (100 MHz, CDCl3): δ 173.4, 171.4, 171.3, 171.0, 149.7, 134.7,126.8, 126.3, 121.9, 117.8, 115.0, 104.5, 80.4, 69.4, 62.2, 61.9, 55.9, 43.9,42.3, 40.6, 40.5, 40.3, 40.0, 34.1, 31.8, 29.2, 29.0, 27.5, 27.2, 25.0, 22.7,19.7, 15.1, 14.2, 13.3.

[0284] Synthesis of compound I-8 (2BL-CASI-C5bMe-2-TG-oleate)

[0285]

[0286] Similar to compound I-7 as described above, compound I-8 was prepared from the intermediate C5bMe-acid-2-TG-oleate (Int-1) as described in Example 1.

[0287] Scheme 5: Synthesis of compound I-9 (2BL-CASI-C5bbGMe-2-TG-octanoate)

[0288]

[0289] At 0°C, thiocarbonate was added dropwise to a solution of Int-3 (500 mg, 1.03 mmol) and potassium carbonate (710 mg, 5.14 mmol) in DMF (10 mL). O -(chloromethyl ester) S A solution of ethyl ester 1 (243 mg, 1.57 mmol, prepared as previously described) in DMF (2 mL) was prepared and the mixture was stirred at room temperature for 18 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 30 mL). The organic fractions were combined and washed with water (90 mL) and brine (90 mL), dried (MgSO4), filtered, and concentrated. The substance was purified by normal phase chromatography (Biotage Isolera, 25 g SiliaSep filter cartridge; eluent 0-25% ethyl acetate / heptane, via 0% 3 CV, 0-25% 13 CV, 25% 1 CV) to give 2 (361 mg, 57% yield) as a colorless oil.

[0290] UPLC4-MS (XB BEH300 C4 20 to 95%): Rt = 5.20 min, 91.6% (UV), 98.3% (ELSD); MS (ESIpos): m / z = 622.7 [M+NH4]+.

[0291] 1 H NMR (400 MHz, CDCl3): δ 5.80 (s, 2H), 5.30 - 5.23 (m, 1H), 4.36 -4.25 (m, 2H), 4.20 - 4.09 (m, 2H), 2.89 (q, J = 7.3 Hz, 2H), 2.55 - 2.41 (m,4H), 2.31 (t, J = 7.5 Hz, 4H), 1.71 - 1.57 (m, 4H), 1.36 - 1.24 (m, 19H), 1.16 - 1.10 (m, 6H), 0.93 - 0.83 (m, 6H).

[0292] Under argon atmosphere, sulfonyl chloride (1 M in DCM, 4.50 mmol, 4.50 mL) was added to a stirred solution of 2 (270 mg, 446 μmol) in DCM (10 mL), and the mixture was heated to 40°C for 3 hours. The mixture was concentrated to give 3 (274 mg, crude substance) as a yellow oil. This substance was used in the next step without any further purification.

[0293] 1 H NMR (400 MHz, CDCl3): δ 5.82 (s, 2H), 5.29 – 5.25 (m, 1H), 4.33 -4.25 (m, 2H), 4.17 – 4.12 (m, 2H), 2.54 (s, 2H), 2.46 (s, 2H), 2.31 (t, J =7.6 Hz, 4H), 1.67 - 1.56 (m, 4H), 1.35 - 1.24 (m, 16H), 1.15 - 1.10 (m, 6H), 0.91 - 0.85 (m, 6H).

[0294] A suspension of 2BL (90.0 mg, 224 μmol), TBAI (43.0 mg, 116 μmol), and NaH (18.0 mg, 450 μmol, 60% w / w dispersion in mineral oil) in tetrahydrofuran (5 mL) was stirred at 0°C for 10 minutes. Tetrahydrofuran (2 mL) containing 4 (260 mg, 449 μmol) was added, and the mixture was gradually warmed to room temperature and stirred for 18 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined fractions were washed with brine (60 mL), dried (MgSO4), filtered, and concentrated. By normal-phase chromatography (Biotage Isolera, 4 g SiliaSep filter cartridge; eluent 50-100% ethyl acetate (+0.5% Et3N) / heptane (+0.5% Et3N), via 2 CV 50%, 20 CV 50-100%, 5 CV 100%), compound I-9 (60.0 mg, 28% yield) was obtained as a yellow gel-like compound, 2BL-CASI-C5bbGMe-2-TG-octanoate.

[0295] UPLC4-MS (XB BEH300 C4 20 to 95%): Rt = 5.36 min, 87.5% (UV), 97.5% (ELSD); MS (ESIpos): m / z = 944.7, 946.6 [M+H]+; 79Br / 81Br isotope mode.

[0296] 1H NMR (400 MHz, CDCl3): δ 7.76 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 7.6Hz, 1H), 7.30 - 7.24 (m, 1H), 6.36 (s, 1H), 6.06 (s, 2H), 5.31 - 5.20 (m,1H), 4.27 (dd, J = 11.9, 4.1 Hz, 2H), 4.11 (m, J = 11.9, 6.0 Hz, 2H), 3.91 -3.82 (m, 1H), 3.53 - 3.32 (m, 5H), 3.21 - 3.11 (m, 1H), 3.04 (dd, J = 11.4,4.9 Hz, 1H), 2.90 - 2.79 (m, 1H), 2.60 (s, 3H), 2.54 (s, 2H), 2.49 - 2.39 (m,3H), 2.34 - 2.25 (m, 4H), 1.65 - 1.55 (m, 4H), 1.34 - 1.22 (m, 19H), 1.20 -1.14 (m, 3H), 1.14 - 1.10 (m, 6H), 0.91 - 0.84 (m, 6H).

[0297] 13 C NMR (100 MHz, CDCl3): δ 173.4, 171.4, 170.8, 170.5, 149.7, 134.64,134.59, 127.7, 126.8, 126.2, 121.9, 120.9, 117.8, 114.9, 104.5, 80.3, 69.1,62.2, 61.9, 55.9, 45.1, 44.5, 44.0, 42.3, 40.5, 40.0, 34.1, 32.8, 31.8, 29.2,29.1, 27.74, 27.65, 27.5, 24.98, 24.95, 22.8, 15.1, 14.2, 13.3。

[0298] Alternative synthetic routes for compound I-9 (2BL-CASI-C5bbGMe-2-TG-octanoate)

[0299] Option 15:

[0300]

[0301] A solution of 4-nitrophenol (3.5 g, 25.2 mmol) and pyridine (2.05 mL, 25.5 mmol) in dichloromethane (25 mL) was added dropwise to a solution of chloromethyl chloroformate (2.5 mL, 28.1 mmol) in dichloromethane (50 mL) over 30 minutes at 0°C. The mixture was stirred at 0°C for 2 hours and then quenched with water (75 mL). The organic layer was washed with 1M sodium hydroxide aqueous solution (30 mL) and brine (50 mL), dried over MgSO4, filtered, and concentrated to give chloromethyl carbonate (4-nitrophenyl ester) (2) as a yellow solid (4.55 g, 72% yield).

[0302] 1 H NMR (400 MHz, CDCl3): δ[ppm] =8.32 - 8.29 (m, 2H), 7.45 - 7.41 (m, 2H), 5.85 (s, 2H).

[0303] A mixture of methyl chlorocarbonate (4-nitrobenzene) (2) (2.5 g, 10.8 mmol), sodium iodide (3.24 g, 21.6 mmol), sodium bicarbonate (181 mg, 2.15 mmol), and acetone (25 mL) was heated at 40°C for 18 hours. The reaction mixture was cooled to ambient temperature, filtered, and washed with acetone (70 mL). The filtrate was concentrated, dissolved in ether (50 mL), and washed with water (50 mL). The organic extract was washed with 10% sodium thiosulfate aqueous solution (50 mL) and brine (50 mL), dried over MgSO4, filtered, and concentrated to give a crude product (3) (2.77 g, 78% yield) as a yellow oil.

[0304] 1 H NMR (400 MHz, CDCl3): δ[ppm] =8.33 – 8.29 (m, 2H), 7.45 – 7.40 (m, 2H), 6.07 (s, 2H).

[0305] A solution of Int-3 (1.6 g, 3.29 mmol) in toluene (5 mL) was added to a suspension of methyl iodide carbonate (4-nitrobenzene ester) (1 g, 3.10 mmol) and silver carbonate (1.2 g, 4.35 mmol) in toluene (15 mL), and stirred at 50°C in the dark for 3 hours under an argon atmosphere. The reaction mixture was quenched with water (60 mL) and extracted into ethyl acetate (3 × 60 mL). The combined organic extracts were washed with brine (150 mL), dried over MgSO4, filtered, and concentrated. The substance was purified by normal-phase chromatography (BioTage Isolera, 25 g Siliasep filter cartridge) using an eluent of 0–25% ethyl acetate / heptane to give the desired product (720 mg, 25% yield) as a white, waxy substance.

[0306] 1 H NMR (400 MHz, CDCl3): δ[ppm] =8.31 ​​– 8.27 (m, 2H), 7.43 – 7.39 (m, 2H), 5.87 (s, 2H), 5.31 – 5.22 (m, 1H), 4.29 (dd, J = 4.2, 11.9 Hz, 2H), 4.14(dd, J = 6.1, 11.9 Hz, 2H), 2.55 (s, 2H), 2.47 (s, 2H), 2.30 (t, J = 7.5 Hz, 4H),1.66 - 1.54 (m, 4H), 1.40 - 1.21 (m, 16H), 1.15 (s, 6H), 0.87 (t, J = 6.8 Hz, 6H).

[0307] At 0°C under an argon atmosphere UnclePotassium butoxide (150 mg, 1.34 mmol) was added to a solution of 2BL (385 mg, 957 µmol) in tetrahydrofuran (THF, 10 mL) and stirred for 30 min. A solution of 1-(1,3-bis(octanoyloxy)propyl-2-yl ester) 5-(((4-nitrophenoxy)carbonyl)oxy)methyl ester) (695 mg, 1.02 mmol) in THF (5 mL) was added to the mixture, and the mixture was gradually heated to room temperature and stirred for 18 h. The reaction mixture was quenched with saturated ammonium chloride aqueous solution (50 mL) and extracted into ethyl acetate (3 × 50 mL). The combined organic extracts were washed with brine (150 mL), dried over MgSO4, filtered, and concentrated. The substance was purified by normal-phase purification (Biotage Isolera, 40g SilliCycle filter cartridge) using 50-100% ethyl acetate + 0.5% Et3N / heptane eluent to obtain a yellow gel-like 2BL-CASI-C5bbGMe-2-TG-octanoate (324 mg, 36% yield).

[0308] 1 H NMR (400 MHz, CDCl3): δ[ppm] = 7.77 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 7.6 Hz, 1H), 7.29 - 7.25 (m, 1H), 6.36 (s, 1H), 6.07 (s, 2H), 5.27 - 5.22(m, 1H), 4.27 (dd, J = 4.2, 12.0 Hz, 2H), 4.12 (dd, J = 6.1, 12.0 Hz, 2H),3.87 - 3.85 (m, 1H), 3.49 - 3.35 (m, 5H), 3.17 - 3.13 (m, 1H), 3.06 - 3.03(m, 1H), 2.87 - 2.83 (m, 1H), 2.60 (br s, 3H), 2.54 (s, 2H), 2.46 - 2.41 (m,3H), 2.29 (t, J = 7.6 Hz, 4H), 1.61 - 1.57 (m, 4H), 1.27 - 1.24 (m, 19H), 1.17 (t, J = 7.1 Hz, 3H), 1.12 (s, 6H), 0.87 (t, J= 6.6 Hz, 6H).

[0309] 13 CNMR (100 MHz, CDCl3): δ [ppm] = 173.4, 170.8, 170.5, 149.7, 134.7, 126.9, 126.2, 121.9, 117.8, 115.0, 80.3, 69.1, 62.2, 61.9, 55.9, 45.1, 44.6,43.9, 42.3, 40.5, 34.1, 32.8, 32.0, 31.8, 29.2, 29.0, 27.7, 25.0, 22.8, 22.7,15.1, 14.3, 14.2, 13.3

[0310] Synthesis of compound I-9' (2BL-CASI-C5bcPr-2-TG-octanoate)

[0311]

[0312] Similar to compound I-9 above, compound I-9' was prepared using intermediate C5bcPr-acid-2-TG-octanoate (Int-5) instead of Int-3.

[0313] Synthesis of compound I-10 (2BL-CASI-C10b'bMe-2-TG-oleate)

[0314]

[0315] Similar to compound I-9 above, compound I-10 was prepared by using intermediate C10b'bMe-acid-2-TG-oleate (Int-19) instead of Int-3.

[0316] Synthesis of compound I-11 (2BL-CMSI-C5bMe-2-TG-oleate)

[0317]

[0318] Similar to compound I-8 above, compound I-11 was prepared by using 1-chloroethyl chloroformate instead of chloromethyl chloroformate in the initial step.

[0319] Scheme 6: Synthesis of compound I-12 (2BL-TML-C5bMe-2-TG-oleate)

[0320]

[0321] Under nitrogen atmosphere, DMAP (97.9 mg, 801 µmol) and EDC-HCl (384 mg, 2.5 mmol) were added to a solution of 1 (310 mg, 1.2 equivalence, 961 µmol) and Int-1 (0.6 g, 1 equivalence, 801 µmol) in DCM (17.8 mL, 0.045 M), and the reaction mixture was stirred at room temperature for two nights. The reaction mixture was diluted with DCM (90 mL) and evaporated onto silica. The crude substance was purified by column chromatography to give 2 (616 mg, 585 µmol, 73%) as a clear oil.

[0322] 1 H NMR (400 MHz, CDCl3): δ 6.80 (dt, J = 2.1, 0.7 Hz, 1H), 6.52 (dt, J= 2.1, 0.7 Hz, 1H), 5.41 – 5.25 (m, 5H), 4.31 (ddd, J = 11.9, 4.3, 2.5 Hz, 2H), 4.14 (dd, J = 11.9, 6.0 Hz, 2H), 3.64 (s, 2H), 3.47 (dd, J = 8.2, 6.9Hz, 2H), 2.68 – 2.42 (m, 7H), 2.39 – 2.26 (m, 5H), 2.23 (d, J = 0.7 Hz, 3H),2.08 – 1.96 (m, 10H), 1.59 (q, J = 6.5 Hz, 6H), 1.41 – 1.23 (m, 47H), 1.13 (d, J = 6.4 Hz, 2H), 0.86 (d, J = 15.4 Hz, 16H), -0.03 (s, 6H).

[0323] C 64 H 112 O9Si [M+NH4] + LRMS (ESI) + Calculated m / z value: 1070.8; Measured value: 1071.

[0324] Under nitrogen atmosphere, in a 20 mL capped vial equipped with a stir bar, 2 (523 mg, 496 µmol) was dissolved in methanol (4.96 mL, 0.1 M) and DCM (4.96 mL, 0.1 M), and then cooled in a water / ice bath. Camphor sulfonic acid (10-CSA, 17.2 mg, 0.15 equivalent, 74.5 µmol) was added in a single addition, and the reaction mixture was stirred for approximately 30 minutes. The ice bath was then removed, and stirring was continued. The reaction mixture was diluted with DCM (10 mL) and washed with saturated sodium bicarbonate aqueous solution (2 × 5 mL). The organic fraction was dried over sodium sulfate, filtered, and evaporated to give a crude substance. This substance was dried, loaded onto silica, and purified by column chromatography (0–30% ethyl acetate / heptane) to give 3 (435 mg, 463 µmol, 93%) as a clear oil.

[0325] 1 H NMR (400 MHz, CDCl3): δ 6.82 (d, J = 2.1 Hz, 1H), 6.53 (d, J = 2.1Hz, 1H), 5.40 – 5.25 (m, 5H), 4.32 (ddd, J = 11.9, 4.3, 1.5 Hz, 2H), 4.15(dd, J = 11.9, 6.0 Hz, 2H), 3.53 (t, J = 7.3 Hz, 2H), 2.72 – 2.44 (m, 7H), 2.42 – 2.26 (m, 5H), 2.23 (s, 3H), 2.10 – 1.94 (m, 9H), 1.66 – 1.37 (m, 12H),1.36 – 1.23 (m, 44H), 1.14 (d, J = 6.4 Hz, 3H), 0.92 – 0.84 (m, 7H).

[0326] C 58 H 98 O9 [M+NH4] + LRMS (ESI) + Calculated m / z value: 956.8; Measured value: 957.1.

[0327] In a 100 mL RBF container equipped with a stir bar, 3 (403 mg, 429 µmol) was dissolved in acetone (6.36 mL, 0.067 M) and cooled to 0°C in a water / ice bath. Jones' reagent (2 M CrO3 in H2SO4) (0.29 mL, 1.34 equivalents, 575 µmol) was added dropwise, and the reaction mixture was stirred at 0°C for four hours. The reaction mixture was quenched with water (5.5 mL) and extracted with ethyl acetate (3 × 5.5 mL). The combined organic extracts were dried over sodium sulfate, filtered, and evaporated to give a crude substance, which was then dried and loaded onto silica. Purification by column chromatography (5–40% ethyl acetate / heptane) yielded 4 (310 mg, 325 µmol, 76%) as a clear oil.

[0328] 1 H NMR (400 MHz, CDCl3): δ 6.82 (d, J = 2.1 Hz, 1H), 6.56 (d, J = 2.0Hz, 1H), 5.41 – 5.23 (m, 5H), 4.31 (dd, J = 11.9, 4.4 Hz, 2H), 4.16 (dd, J =11.9, 5.9 Hz, 2H), 2.82 (d, J = 1.3 Hz, 2H), 2.67 (dd, J = 15.5, 5.9 Hz, 1H), 2.64 – 2.47 (m, 6H), 2.37 (dd, J = 15.2, 7.1 Hz, 1H), 2.31 (td, J = 7.6, 1.7Hz, 4H), 2.23 (s, 3H), 2.04 – 1.97 (m, 8H), 1.63 – 1.55 (m, 11H), 1.36 – 1.22(m, 47H), 1.13 (d, J = 6.6 Hz, 3H), 0.94 – 0.84 (m, 6H).

[0329] C 58 H 96 O 10 [M+NH4] + LRMS (ESI) + Calculated m / z value: 970.7; Measured value: 971.0.

[0330] Oxaloyl chloride (1.5 equivalents) and DMF (1-2 drops, catalytic amount) were added to DCM (0.1 M) containing 4 (1 equivalent) at 0 °C, and the resulting mixture was stirred at 0 °C for 1 to 2 hours. The reactants were concentrated under reduced pressure, and the resulting residue was redistilled from toluene 3 to 4 times to give 5, which was used for the next step without further purification.

[0331] At 0 °C, a solution of 2BL (1 equivalent) in THF (0.1 M) was added to a suspension of NaH (60% w / w dispersion in mineral oil, 1 equivalent) in THF (0.1 M), and the reaction mixture was stirred at 0 °C for about 30 min. A solution of 5 (1.2 equivalent) in THF (0.1 M) was slowly added, and the resulting mixture was stirred at 0 °C for about 30 min, then warmed to room temperature and stirred at room temperature for 4 to 24 h. The reaction mixture was quenched by slow addition of a saturated aqueous NH4Cl solution, diluted with water, and extracted with ethyl acetate. The combined organic extracts were washed with water and brine, dried over MgSO4, and concentrated under reduced pressure to give a crude reaction mixture. Purification by silica gel chromatography with a suitable solvent mixture yielded 2BL-TML-C5bMe-2-TG-oleate, compound I-12.

[0332] Synthesis of compound 1-13 (2BL-TML-C8bMe-2-TG-oleate)

[0333]

[0334] Similar to compound I-12 as described above, compound I-13 was prepared from the intermediate C8bMe-acid-2-TG-oleate (Int-16) as described in Example 1.

[0335] Scheme 7: Synthesis of compound I-14 (2BL-FSI4-C5bMe-2-TG-oleate)

[0336]

[0337] TBAI (494 mg, 1.34 mmol) and cesium carbonate (2.61 g, 8.01 mmol) were added to a stirred solution of Int-1 (2.00 g, 2.67 mmol) and tert-butyl 4-bromobutyrate 1 (596 mg, 2.67 mmol, 474 μL) in toluene (10 mL) at room temperature. The reaction mixture was heated to 80°C and stirred for 4 hours. The reaction mixture was diluted with ethyl acetate, and the organic matter was washed with brine, dried over sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by normal-phase purification (Biotage Isolera, 80 g SiliaSep filter cartridge; eluent 0-30% ethyl acetate / heptane) to give 2 (1.66 g, 69.8% yield) as a colorless oil.

[0338] ¹H NMR (400 MHz, CDCl3): δ 5.39 - 5.24 (m, 5H), 4.30 (ddd, J = 3.2,5.7, 9.3 Hz, 2H), 4.16 - 4.08 (m, 4H), 2.51 - 2.35 (m, 3H), 2.33 - 2.19 (m,8H), 2.06 - 1.96 (m, 8H), 1.95 - 1.87 (m, 2H), 1.65 - 1.57 (m, 4H), 1.44 (s,9H), 1.37 - 1.20 (m, 40H), 1.02 (d, J = 6.7 Hz, 3H), 0.88 (t, J = 6.6 Hz, 6H).

[0339] 13 C NMR (101 MHz, CDCl3): δ 173.4, 172.3, 172.2, 171.5, 130.2, 129.9,80.6, 69.2, 63.6, 62.2, 40.82, 40.78, 34.1, 32.08, 32.05, 29.91, 29.85, 29.7,29.5, 29.32, 29.26, 29.2, 28.2, 27.5, 27.4, 27.3, 25.0, 24.3, 22.8, 19.8,14.3.

[0340] Trifluoroacetic acid (TFA, 2.98 g, 26.1 mmol, 2.00 mL) was added to a stirred solution of 2 (500 mg, 561 μmol) in DCM (8 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate (50 mL), washed with aqueous sodium bicarbonate solution (3 × 50 mL), followed by water (2 × 50 mL), and then brine (3 × 50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum to give 3 (405 mg, 86.5% yield) as a colorless oil.

[0341] UPLC4-MS (XB BEH 300 C4 20-95%): Rt = 6.30 min, 60.7% (UV), 93.8% (ELSD). MS (ESIpos): m / z = 853.0 [M+NH4]+.

[0342] ¹H NMR (400 MHz, CDCl₃): δ 5.55 - 5.00 (m, 5H), 4.29 (dd, J = 4.2, 11.9 Hz, 2H), 4.18 - 4.06 (m, 4H), 2.51 - 2.35 (m, 5H), 2.33 - 2.19 (m, 6H), 2.16 - 1.77 (m, 10H), 1.64 - 1.55 (m, 4H), 1.27 (m, 40H), 1.01 (d, J = 6.4 Hz, 3H), 0.86 (br t, J = 6.7 Hz, 6H); No exchangeable CO₂H protons were observed.

[0343] 13 C NMR (101 MHz, CDCl3): δ 173.46, 173.44, 172.2, 171.6, 130.2,129.9, 69.3, 63.4, 62.2, 40.8, 40.7, 34.1, 32.0, 30.6, 29.9, 29.83, 29.79,29.7, 29.5, 29.4, 29.3, 29.25, 29.22, 27.5, 27.4, 27.3, 25.0, 24.0, 22.8,19.8, 14.2.

[0344] Oxaloyl chloride (76.0 mg, 599 μmol, 50.7 μL) and DMF (3 drops, catalytic amount) were added to a stirred solution of 3 (400 mg, 479 μmol) in DCM (8 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum to give 4 (406 mg, 88.4% yield) as a yellow oil.

[0345] ¹H NMR (400 MHz, CDCl3): δ 5.64 - 5.01 (m, 5H), 4.37 - 4.23 (m, 2H), 4.21 - 4.05 (m, 4H), 3.00 - 2.95 (m, 3H), 2.52 - 2.35 (m, 3H), 2.35 - 2.18(m, 6H), 2.08 - 1.95 (m, 9H), 1.60 (br t, J = 6.8 Hz, 4H), 1.37 - 1.20 (m,40H), 1.02 (d, J = 6.4 Hz, 3H), 0.87 (t, J = 6.7 Hz, 6H).

[0346] NaH (12.0 mg, 300 μmol, 60% purity) and TBAI (44.0 mg, 119 μmol) were added to a 2BL (94.0 mg, 234 μmol) solution in tetrahydrofuran (4 mL) under stirring at 0°C. The solution was stirred at 0°C for 10 minutes. Then, tetrahydrofuran (4 mL) containing 4 (400 mg, 469 μmol) was added at 0°C. The solution was warmed to room temperature and stirred for 3 hours. The reaction mixture was diluted with ethyl acetate (50 mL), washed with water (50 mL), and then washed with brine (50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum to give a crude yellow oil. The substance was purified by normal phase chromatography (Biotage Isolera, 25 g SiliCycle filter cartridge; eluent 0-80% ethyl acetate + 0.5% Et3N / heptane + 0.5% Et3N, 30 CVs) to give 2BL-FSI4-C5bMe-2-TG-oleate, compound I-14 (147 mg, 51.3% yield), which was a yellow oil.

[0347] UPLC4-MS (XB BEH 300 C4 20-95%): Rt = 7.10 min, 78.4% (UV), 92.3% (ELSD). MS (ESIpos): m / z = 1219.0, 1220.9 [M+H]+ 79Br / 81Br isotope pair.

[0348] ¹H NMR (400 MHz, CDCl3): δ 7.98 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 7.6Hz, 1H), 7.32 - 7.23 (m, 1H), 6.37 (s, 1H), 5.53 - 5.01 (m, 5H), 4.34 - 4.18(m, 4H), 4.13 (br dd, J = 5.9, 11.9 Hz, 2H), 3.88 (br s, 1H), 3.51 - 3.32 (m,5H), 3.31 - 3.14 (m, 3H), 3.06 (br dd, J = 4.2, 10.9 Hz, 1H), 2.90 - 2.74 (m,1H), 2.61 (s, 3H), 2.52 - 2.35 (m, 4H), 2.33 - 2.15 (m, 8H), 2.14 - 1.79 (m,8H), 1.64 - 1.55 (m, 4H), 1.27 (m, 43H), 1.17 (t, J = 7.1 Hz, 3H), 1.02 (d, J= 6.4 Hz, 3H), 0.87 (t, J = 6.6 Hz, 6H).

[0349] 13 C NMR (101 MHz, CDCl3): δ 173.3, 172.2, 171.8, 171.4, 171.3, 135.6,134.5, 130.1, 129.8, 128.2, 127.4, 126.52, 126.47, 121.8, 121.0, 117.8,115.5, 103.1, 69.3, 63.5, 62.2, 62.0, 55.9, 43.9, 42.2, 40.77, 40.74, 39.9,35.7, 34.1, 32.0, 29.9, 29.82, 29.77, 29.7, 29.6, 29.4, 29.3, 29.23, 29.21,27.7, 27.5, 27.4, 27.34, 27.29, 24.9, 24.2, 22.8, 19.8, 15.1, 14.2, 13.3。

[0350] Scheme 8: Synthesis of compound I-15 (2BL-FSI4-C5bbGMe-2-TG-octanoate)

[0351]

[0352] 1-tert-butyl 4-bromobutyrate (344 mg, 1.54 mmol, 273 μL) was added to a stirred solution of Int-3 (750 mg, 1.54 mmol), cesium carbonate (1.51 g, 4.62 mmol), and TBAI (285 mg, 772 μmol) in toluene (15 mL). The reaction mixture was heated to 80°C and stirred for 4 hours. The reaction mixture was cooled and diluted with ethyl acetate (100 mL), and washed with water (100 mL) and brine (2 × 100 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give a crude yellow oil. This substance was purified by normal-phase purification (Biotage Isolera, 80 g SiliCycle filter cartridge; eluent 0-30% ethyl acetate / heptane, 24 CVs) to give 2 (505 mg, 52% yield) as a colorless oil.

[0353] UPLC4-MS (XB BEH 300 C4 20-95%): Rt = 5.35 min, 97.2% (UV), 99.5% (ELSD). MS (ESIpos): m / z = 647.2 [M+NH4]+.

[0354] ¹H NMR (400 MHz, CDCl3): δ 5.35 - 5.15 (m, 1H), 4.33 - 4.22 (m, 2H), 4.13 (dd, J = 6.1, 11.9 Hz, 2H), 4.07 (t, J = 6.5 Hz, 2H), 2.44 (s, 2H), 2.40(s, 2H), 2.34 - 2.10 (m, 6H), 1.91 (quin, J = 6.9 Hz, 2H), 1.64 - 1.55 (m,4H), 1.44 (s, 9H), 1.27 (m, 16H), 1.11 (s, 6H), 0.87 (t, J = 6.4 Hz, 6H).

[0355] 13 C NMR (101 MHz, CDCl3): δ 173.38, 172.2, 171.71, 171.69, 171.0, 80.59, 69.0, 63.3, 62.3, 45.3, 34.1, 32.7, 32.1, 31.8, 29.2, 29.0, 28.2,27.6, 24.9, 24.3, 22.7, 14.18.

[0356] TFA (3.00 g, 26.3 mmol, 2.00 mL) was added to a solution of 2 (500 mg, 795 μmol) in DCM (8 mL) at room temperature. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with ethyl acetate (50 mL), washed with aqueous sodium bicarbonate solution (3 × 50 mL), then with water (2 × 50 mL) and brine (3 × 50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give 3 (384 mg, 84% yield) as a colorless oil.

[0357] UPLC4-MS (XB BEH 300 C4 20-95%): Rt = 3.61 min, 96.3% (UV), 97.5% (ELSD). MS (ESIpos): m / z = 590.7 [M+NH4]+.

[0358] ¹H NMR (400 MHz, CDCl₃): δ 5.34 – 5.20 (m, 1H), 4.29 (dd, J = 4.3, 11.9 Hz, 2H), 4.21 – 4.04 (m, 4H), 2.48 – 2.42 (m, 4H), 2.40 (s, 2H), 2.30 (t, J = 7.6 Hz, 4H), 1.97 (quin, J = 6.8 Hz, 2H), 1.60 (m, 4H), 1.33 – 1.22 (m, 16H), 1.11 (s, 6H), 0.87 (t, J = 6.5 Hz, 6H); No exchangeable CO₂H protons were observed.

[0359] 13 C NMR (101 MHz, CDCl3): δ 173.5, 171.7, 171.0, 69.0, 63.1, 62.3,45.2, 45.1, 34.2, 32.8, 31.8, 30.6, 29.2, 29.0, 27.7, 25.0, 23.9, 22.7, 14.2.

[0360] Oxaloyl chloride (103 mg, 814 μmol, 71.0 μL) and DMF (3 drops, catalytic amount) were added to a solution of 3 (370 mg, 646 μmol) in DCM (7 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give 4 (379 mg, 99% yield) as a yellow oil.

[0361] ¹H NMR (400 MHz, CDCl3): δ 5.31 - 5.20 (m, 1H), 4.28 (dd, J = 4.3,11.9 Hz, 2H), 4.19 - 4.00 (m, 4H), 2.99 (t, J = 7.2 Hz, 2H), 2.44 (s, 2H),2.41 (s, 2H), 2.30 (t, J = 7.5 Hz, 4H), 2.03 (quin, J = 6.7 Hz, 2H), 1.64 -1.55 (m, 4H), 1.33 - 1.22 (m, 16H), 1.11 (s, 6H), 0.87 (t, J = 6.7 Hz, 6H).

[0362] 13 C NMR (101 MHz, CDCl3): δ 173.39, 173.36, 171.6, 170.9, 69.1, 62.23,62.18, 45.1, 45.0, 43.9, 34.1, 32.7, 31.8, 29.2, 29.0, 27.7, 25.0, 24.4,22.7, 14.9.

[0363] Sodium hydride (16.0 mg, 400 μmol, 60% w / w dispersion in mineral oil) and TBAI (59.0 mg, 160 μmol) were added to a 2BL (128 mg, 318 μmol) solution in tetrahydrofuran (4 mL) under stirring at 0°C. The solution was stirred at 0°C for 10 minutes. Then, tetrahydrofuran (4 mL) containing 4 (375 mg, 634 μmol) was added at 0°C. The solution was warmed to room temperature and stirred for 3 hours. The reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give a crude yellow oil. The substance was purified by normal-phase purification (Biotage Isolera, 25 g SiliCycle filter cartridge; eluent 0-80% ethyl acetate (+0.5% Et3N) / heptane (+0.5% Et3N), 24 CVs) to give 2BL-FSI4-C5bbGMe-2-TG-octanoate, compound I-15 (185 mg, 61% yield), which is a yellow oil.

[0364] UPLC4-MS: (XB BEH 300 C4 20 - 95%): Rt = 5.43 min, 96.2% (UV), 100.0% (ELSD). MS (ESIpos): m / z = 956.8, 958.7 [M+H]+; 79Br / 81Br isotope pair.

[0365] ¹H NMR (400 MHz, CDCl3): δ 7.96 (d, J = 8.1 Hz, 1H), 7.36 (d, J = 7.6Hz, 1H), 7.30 - 7.22 (m, 1H), 6.35 (s, 1H), 5.28 - 5.20 (m, 1H), 4.26 (dd, J= 4.2, 11.9 Hz, 2H), 4.20 (t, J = 6.3 Hz, 2H), 4.12 (dd, J = 6.1, 11.9 Hz,2H), 3.87 (br d, J = 4.5 Hz, 1H), 3.50 - 3.30 (m, 5H), 3.29 - 3.13 (m, 3H),3.05 (br dd, J = 4.7, 11.3 Hz, 1H), 2.91 - 2.79 (m, 1H), 2.60 (s, 3H), 2.50 -2.42 (m, 3H), 2.39 (s, 2H), 2.28 (t, J = 7.5 Hz, 4H), 2.17 (quin, J = 6.7 Hz,2H), 1.62-1.54 (m, 4H), 1.30 - 1.21 (m, 19H), 1.16 (t, J = 7.1 Hz, 3H), 1.09(s, 6H), 0.85 (t, J = 6.7 Hz, 6H).

[0366] 13C NMR (101 MHz, CDCl3): δ 173.4, 171.9, 171.7, 171.3, 170.9, 135.5,134.5, 127.3, 126.5, 126.4, 121.7, 121.0, 117.8, 115.5, 103.1, 69.0, 63.3,62.2, 61.9, 55.8, 45.2, 43.9, 42.2, 40.5, 39.9, 35.8, 34.1, 32.7, 31.7, 29.2,29.0, 27.6, 24.9, 24.2, 22.7, 15.1, 14.2, 13.3.

[0367] Synthesis of compound I-15' (2BL-FSI4-C5bcPr-2-TG-octanoate)

[0368]

[0369] Similar to compound I-15 mentioned above, compound I-15' was prepared using the intermediate C5bcPr-acid-2-TG-octanoate (Int-5) instead of Int-3.

[0370] Scheme 9: Synthesis of compound I-16 (2BL-FSI5-C5bbGMe-2-TG-octanoate)

[0371]

[0372] NaH (89.5 mg, 2.24 mmol, 60% w / w dispersion in mineral oil) was added to a stirred solution of 2 BL (600 mg, 1.49 mmol) in DMF (10 mL) at 0°C under argon atmosphere. The mixture was stirred for 30 min while maintaining the temperature at 0°C. Then, 1 bromopentanoyl chloride (446 mg, 2.24 mmol, 299 μL) was added, and the mixture was warmed to room temperature and stirred for 18 h. The reaction mixture was quenched with a saturated aqueous solution of NH4Cl (50 mL), and the mixture was extracted with diethyl ether (3 × 50 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, filtered, and concentrated to give crude oil. The substance was purified by normal phase chromatography (Biotage Isolera, 40 g SiliaSep filter cartridge; eluent 0-10% methanol / DCM, 30 CVs) to give 2 (505 mg, 30% yield) as a yellow oil.

[0373] UPLC6 (BEH C18 long neutral 2 to 95): Rt = 2.73 min, 51.3% (UV). MS (ESIpos): m / z = 566.0, 568.0 [M+H]+.

[0374] 1 H NMR (400 MHz, CDCl3) δ 7.97 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 7.6Hz, 1H), 7.31 - 7.23 (m, 1H), 6.40 - 6.35 (m, 1H), 3.92 (br s, 1H), 3.61 (t,J = 6.3 Hz, 1H), 3.56 - 3.31 (m, 4H), 3.21 (br t, J = 6.2 Hz, 2H), 3.12 (brd, J = 4.4 Hz, 1H), 2.97 - 2.85 (m, 1H), 2.63 (br s, 3H), 2.60 - 2.46 (m,2H), 2.42 - 2.31 (m, 2H), 2.04 - 1.86 (m, 2H), 1.84 - 1.74 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H), 1.17 (t, J = 7.1 Hz, 3H); As observed by UPLC, the product contains 5.74% w / w DCM and other impurities.

[0375] At room temperature, cesium carbonate (178 mg, 548 μmol) and TBAI (33.8 mg, 91.4 μmol) were added to a stirred solution of 2 (120 mg, 183 μmol) and Int-3 (115 mg, 237 μmol) in toluene (4 mL). The reaction mixture was heated to 50°C and stirred for 2 hours. The reaction mixture was diluted with ethyl acetate (25 mL), washed with water (25 mL) and brine (25 mL), dried over sodium sulfate, filtered, and concentrated to give a crude yellow oil. The substance was purified by normal phase chromatography (Biotage Isolera, 25 g Silicycle filter cartridge; eluent 0-100% ethyl acetate / heptane, 30 CVs) to give 2BL-FSI5-C5bbGMe-2-TG-octanoate, compound I-16 (40.0 mg, 23% yield), as a yellow oil.

[0376] UPLC4 (XB BEH 300 C4 20 to 95): Rt = 5.54 min, 92.8% (UV), 99.9% (ELSD). MS (ESIpos): m / z = 970.8, 972.7 [M+H]+ 79Br / 81Br isotope pair.

[0377] 11H NMR (400 MHz, CDCl3): δ 7.96 (d, J = 8.2 Hz, 1H), 7.36 (d, J = 7.6Hz, 1H), 7.26 (t, J = 8.0 Hz, 1H), 6.36 (s, 1H), 5.29 - 5.23 (m, 1H), 4.27(dd, J = 4.3, 11.9 Hz, 2H), 4.17 - 4.08 (m, 4H), 3.88 (br s, 1H), 3.50 - 3.39(m, 4H), 3.38 - 3.30 (m, 1H), 3.25 - 3.12 (m, 3H), 3.05 (br dd, J = 4.7, 11.2Hz, 1H), 2.86 (br t, J = 10.9 Hz, 1H), 2.60 (s, 3H), 2.53 - 2.42 (m, 3H),2.39 (s, 2H), 2.29 (t, J = 7.6 Hz, 4H), 1.97 - 1.85 (m, 2H), 1.84 - 1.72 (m,2H), 1.64 - 1.53 (m, 4H), 1.33 - 1.21 (m, 19H), 1.16 (t, J = 7.1 Hz, 3H),1.11 (s, 6H), 0.86 (t, J = 6.7 Hz, 6H); 76H expected, 76H observed.

[0378] 13 13C NMR (101 MHz, CDCl3): δ 173.4, 172.3, 171.8, 171.3, 171.0, 135.5,134.6, 127.4, 126.5, 121.7, 120.9, 117.7, 115.4, 103.1, 69.0, 63.9, 62.2,62.0, 55.9, 45.3, 45.2, 43.9, 42.2, 40.5, 39.9, 38.6, 34.1, 32.7, 31.8, 29.2,29.0, 28.2, 27.6, 24.9, 22.7, 21.7, 15.1, 14.2, 13.3.

[0379] Scheme 10: Synthesis of compound I-17 (2BL-FSI5-C5bMe-2-TG-oleate)

[0380]

[0381] Sodium hydride (89.5 mg, 2.24 mmol, 60% w / w dispersion in mineral oil) was added to a stirred solution of 2BL (600 mg, 1.49 mmol) in DMF (10 mL) at 0°C under argon atmosphere. The mixture was stirred for 30 minutes while maintaining the temperature. Then 5-bromopentanoyl chloride 1 (446 mg, 2.24 mmol, 299 μL) was added, and the mixture was warmed to room temperature and stirred for 4 hours. The substance was diluted with ether (20 mL) and washed with water (2 × 10 mL), dried over MgSO4, filtered, and concentrated to give crude oil. The crude substance was purified by normal phase chromatography (Biotage Isolera, 40 g SiliaSep filter cartridge; eluent 0-10% methanol / DCM, 30 CVs) to give 2 (553 mg, 49.2% yield) as a yellow oil.

[0382] C_UPLC2-MS: (CSH-C18 long neutral 2 to 95%) Rt = 1.91 min (75.4%), MS (ESIpos): m / z = [M+H]+ 566, 568.2.

[0383] 1 H NMR (400 MHz, CDCl3): δ 7.40 - 7.35 (m, 1H), 7.32 - 7.28 (m, 1H), 7.27 - 7.25 (m, 1H), 6.38 (s, 1H), 3.61 (t, J = 6.2 Hz, 1H), 3.54 (br t, J =6.3 Hz, 1H), 3.50 - 3.31 (m, 2H), 3.21 (br t, J = 6.1 Hz, 3H), 2.92 - 2.82(m, 2H), 2.62 (br s, 2H), 2.52 - 2.32 (m, 2H), 2.11 - 1.98 (m, 4H), 1.98 -1.82 (m, 2H), 1.82 - 1.73 (m, 2H), 1.26 (br t, J = 7.0 Hz, 3H), 1.17 (t, J =7.1 Hz, 3H).

[0384] At room temperature, cesium carbonate (644 mg, 1.98 mmol) and TBAI (127 mg, 343 μmol) were added to a solution of Int-1 (585 mg, 782 μmol) and 2 (553 mg, 651 μmol) in toluene (20 mL). The reaction mixture was heated to 50°C and stirred for 3 hours. The reaction mixture was quenched with saturated NH4Cl solution (100 mL), and the organic matter was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, and concentrated under vacuum to give a crude residue. The residue was purified by normal-phase purification (Biotage Isolera, 40 g SiliCycle filter cartridge; eluent 0-100% ethyl acetate / heptane, 40 CVs) to give 2BL-FSI5-C5bMe-2-TG-oleate, compound I-17 (429 mg, 53.4% ​​yield), which was a yellow oil.

[0385] UPLC4-MS (XB BEH300 C4 20 to 95%): Rt = 7.15 min, 78.1% (UV), 93.6% (ELSD); MS (ESIpos): m / z = [M+H]+ 1232.96, 1234.91.

[0386] 1H NMR (400 MHz, CDCl3): δ 7.97 (d, J = 8.1 Hz, 1H), 7.37 (d, J = 7.6Hz, 1H), 7.30 - 7.26 (m, 1H), 6.37 (s, 1H), 5.38 - 5.24 (m, 5H), 4.32 - 4.26(m, 2H), 4.18 - 4.10 (m, 4H), 3.85 (br s, 1H), 3.51 - 3.40 (m, 4H), 3.36 (brdd, J = 5.7, 15.5 Hz, 1H), 3.26 - 3.14 (m, 3H), 3.09 - 3.01 (m, 1H), 2.89 -2.82 (m, 1H), 2.60 (s, 3H), 2.49 - 2.34 (m, 4H), 2.34 - 2.20 (m, 6H), 2.05 -1.95 (m, 8H), 1.95 - 1.88 (m, 2H), 1.88 - 1.72 (m, 2H), 1.65 - 1.54 (m, 4H),1.28 (br d, J = 12.8 Hz, 44H), 1.17 (t, J = 7.1 Hz, 3H), 1.02 (d, J = 6.5 Hz,3H), 0.92 - 0.83 (m, 6H).

[0387] 13 C NMR (100 MHz, CDCl3): δ 173.4, 172.4, 171.5, 171.4,8135.6, 134.7,130.2, 129.9, 127.5, 126.52, 126.48, 121.8, 121.0, 117.7, 115.4, 103.1, 69.3,64.2, 62.2, 62.1, 60.5, 56.0, 44.0, 42.3, 40.8, 40.5, 40.1, 38.6, 34.2, 32.1,29.93, 29.87, 29.7, 29.5, 29.34, 29.28, 28.2, 27.8, 27.5, 27.4, 27.3, 25.0,22.8, 21.7, 21.2, 19.8, 15.2, 14.4, 14.3, 13.3。

[0388] Scheme 11: Synthesis of compound I-18 (2BL-FSI5-C5cPr-2-TG-octanoate)

[0389]

[0390] Sodium hydride (22.4 mg, 559 μmol, 60% w / w dispersion in mineral oil) was added to a stirred solution of 2BL (150 mg, 373 μmol) in tetrahydrofuran (5 mL), and the mixture was stirred at room temperature for 10 min. 5-Bromopentanoyl chloride 1 (112 mg, 559 μmol, 74.9 μL) was added to the solution, and the reaction mixture was stirred at room temperature for 18 h. The substance was diluted with DCM (20 mL), washed with water (10 mL) and brine (10 mL), dried over MgSO4, filtered, and concentrated to give a crude oil. The substance was purified by normal-phase chromatography (Biotage Isolera, 12 g SiliaSep filter cartridge; eluent 0–30% (10% methanol / DCM) / DCM, 30 CVs) to give 2 (150 mg, 61% yield) as a yellow oil.

[0391] UPLC6-MS: (BEH-C18 short bases 2 to 95%) Rt = 1.13 min, 86.2% (UV), MS (ESIpos): m / z = [M+H]+ 566.1, 568.1 79Br / 81Br isotope pair.

[0392] ¹H NMR (400 MHz, CDCl3): δ 7.97 (d, J = 8.1 Hz, 1H), 7.37 (d, J = 7.3Hz, 1H), 7.29 (d, J = 8.1 Hz, 1H), 6.39 - 6.35 (m, 1H), 4.06 - 3.85 (m, 1H),3.58 - 3.29 (m, 7H), 3.28 - 3.17 (m, 2H), 3.09 (br dd, J = 4.0, 10.6 Hz, 1H),3.01 - 2.81 (m, 1H), 2.75 - 2.48 (m, 4H), 2.11 - 1.92 (m, 4H), 1.79 (br dd, J= 2.0, 4.8 Hz, 1H), 1.30 - 1.22 (m, 3H), 1.17 (t, J = 7.1 Hz, 3H).

[0393] At room temperature, cesium carbonate (223 mg, 684 μmol) and TBAI (43.0 mg, 114 μmol) were added to a solution of 2 (150 mg, 228 μmol) and -5 (111 mg, 229 μmol) in toluene (4 mL). The reaction mixture was heated to 50°C and stirred for 2 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (25 mL), and washed with water (25 mL) and brine (25 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give a crude yellow oil. The substance was purified by normal phase chromatography (Biotage Isolera, 25 g SiliaSep filter cartridge; eluent 0-80% ethyl acetate (+0.5% Et3N) / heptane (+0.5% Et3N), 30 CVs) to give 2BL-FSI5-C5bcPr-2-TG-octanoate, compound I-18 (50.0 mg, 23% yield), as a yellow oil.

[0394] UPLC4-MS: (XB BEH 300 C4 20 to 95%): Rt = 5.38 min, 89.8% (UV), 100.0% (ELSD). MS (ESIpos): [M+H]+ m / z = 968.8, 970.8 79Br / 81Br isotope pair.

[0395] ¹H NMR (400 MHz, CDCl3): δ 7.95 (d, J = 8.1 Hz, 1H), 7.35 (d, J = 7.5Hz, 1H), 7.26 (t, J = 6.2 Hz, 1H), 6.35 (s, 1H), 5.29 - 5.20 (m, 1H), 4.29(dd, J = 4.4, 11.9 Hz, 2H), 4.16 - 4.07 (m, 4H), 3.89 (br s, 1H), 3.48 - 3.31(m, 5H), 3.19 (br t, J = 7.2 Hz, 3H), 3.05 (br dd, J = 4.6, 11.1 Hz, 1H),2.92 - 2.79 (m, 1H), 2.61 (s, 3H), 2.51 - 2.45 (m, 1H), 2.42 (s, 2H), 2.38(s, 2H), 2.29 (t, J = 7.6 Hz, 4H), 1.96 - 1.85 (m, 2H), 1.81 - 1.73 (m, 2H),1.58 (quin, J = 7.3 Hz, 4H), 1.32 - 1.19 (m, 19H), 1.15 (t, J = 7.1 Hz, 3H),0.85 (t, J = 6.4 Hz, 6H), 0.53 (m, 4H).

[0396] 13 C NMR (100 MHz, CDCl3): δ 173.3, 172.3, 172.2, 171.33, 171.25,135.5, 134.5, 127.3, 126.4, 121.7, 120.8, 117.6, 115.4, 103.1, 69.1, 64.0,62.1, 62.0, 55.9, 43.8, 42.2, 41.07, 41.05, 40.5, 39.9, 38.6, 34.1, 31.7,29.2, 29.0, 28.2, 27.6, 24.9, 22.7, 21.6, 15.1, 14.4, 14.2, 13.3, 12.3。

[0397] Scheme 12: Synthesis of compound I-19 (2BL-FSI5-C6bbGMe-2-TG-octanoate)

[0398]

[0399] NaH (89.5 mg, 2.24 mmol, 60% w / w dispersion in mineral oil) was added to a stirred solution of 2 BL (600 mg, 1.49 mmol) in DMF (10 mL) at 0°C under argon atmosphere. The mixture was stirred for 30 min while maintaining the temperature at 0°C. Then, 5-bromopentanoyl chloride (446 mg, 2.24 mmol, 299 μL) was added, and the mixture was warmed to room temperature and stirred for 18 h. The reaction mixture was quenched with a saturated aqueous solution of NH4Cl (50 mL), and the mixture was extracted with diethyl ether (3 × 50 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, filtered, and concentrated to give crude oil. The substance was purified by normal phase chromatography (Biotage Isolera, 40 g SiliaSep filter cartridge; eluent 0-10% methanol / DCM, 30 CVs) to give 2 (505 mg, 30% yield) as a yellow oil.

[0400] UPLC6 (BEH C18 long neutral 20 to 95): Rt = 2.73 min, 51.3% (UV). MS (ESIpos): m / z = 566.0, 568.0 [M+H]+.

[0401] 1 H NMR (400 MHz, CDCl3) δ 7.97 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 7.6Hz, 1H), 7.31 - 7.23 (m, 1H), 6.40 - 6.35 (m, 1H), 3.92 (br s, 1H), 3.61 (t,J = 6.3 Hz, 1H), 3.56 - 3.31 (m, 4H), 3.21 (br t, J = 6.2 Hz, 2H), 3.12 (brd, J = 4.4 Hz, 1H), 2.97 - 2.85 (m, 1H), 2.63 (br s, 3H), 2.60 - 2.46 (m,2H), 2.42 - 2.31 (m, 2H), 2.04 - 1.86 (m, 2H), 1.84 - 1.74 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H), 1.17 (t, J = 7.1 Hz, 3H); As observed by UPLC, the product contains 5.74% w / w DCM and other impurities.

[0402] At room temperature, cesium carbonate (291 mg, 894 μmol) and TBAI (57.2 mg, 155 μmol) were added to a solution of Int-6 (147 mg, 294 μmol) and 2 (250 mg, 294 μmol) in toluene (20 mL). The reaction mixture was heated to 50°C and stirred for 3 hours. The reaction mixture was quenched with saturated aqueous NH4Cl solution (100 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (100 mL), dried over MgSO4, and concentrated to give a crude residue. The residue was purified by normal-phase chromatography (Biotage Isolera, 40 g SiliaSep filter cartridge; eluent 0-10% methanol / DCM, 15 CVs) to give 2BL-FSI5-C6bbGMe-2-TG-octanoate, compound I-19 (88.0 mg, 30% yield), as a yellow oil.

[0403] UPLC-MS (XB BEH300 C4 20 to 95%): Rt = 5.58 min, 66.7% (UV), 93.2% (ELSD); MS (ESIpos): m / z = 986.7, 987.8 [M+H]+. 79Br / 81Br isotope pair.

[0404] 1H NMR (400 MHz, CDCl3): δ 7.95 (d, J = 8.2 Hz, 1H), 7.36 (d, J = 7.6Hz, 1H), 7.26 (t, J = 7.7 Hz, 1H), 6.35 (s, 1H), 5.29 - 5.23 (m, 1H), 4.31 -4.24 (m, 2H), 4.17 - 4.08 (m, 4H), 3.88 - 3.82 (m, 1H), 3.52 - 3.32 (m, 6H), 3.24 - 3.12 (m, 3H), 3.07 - 2.99 (m, 1H), 2.88 - 2.80 (m, 1H), 2.59 (s, 3H), 2.32 - 2.26 (m, 6H), 2.21 (s, 2H), 1.99 - 1.85 (m, 2H), 1.85 - 1.74 (m, 2H), 1.70 - 1.55 (m, 6H), 1.30 - 1.22 (m, 19H), 1.16 (t, J = 7.1 Hz, 3H), 0.99 (s, 6H), 0.89 - 0.83 (m, 6H); The product contains a small amount of impurities.

[0405] 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.4, 172.4, 171.4, 171.0, 135.5,134.6, 127.4, 126.5, 121.7, 121.0, 117.7, 115.4, 103.1, 69.0, 64.2, 62.3,62.0, 55.9, 45.7, 43.9, 42.2, 40.5, 40.0, 38.6, 36.9, 34.1, 33.1, 31.8, 29.6,29.2, 29.0, 28.2, 27.7, 27.3, 27.0, 24.9, 22.7, 21.6, 15.1, 14.2, 13.3.

[0406] Scheme 13: Synthesis of compound I-20 (2BL-PHB-C5bMe-2-TG-oleate)

[0407]

[0408] 4-Hydroxybenzaldehyde 1 (277 mg, 2.27 mmol, 245 µL) and DMAP (555 mg, 4.55 mmol) were added to a solution of Int-1 (1.70 g, 2.27 mmol) and EDC-HCl (870 mg, 4.54 mmol) in DCM (30 mL), and the mixture was stirred at room temperature for about 18 hours. The substance was purified by normal phase chromatography (BioTage Isolera, 40 g SiliaSep filter cartridge) using 0-30% ethyl acetate eluent / heptane for 20 CV steps to give (2) as a colorless oil (1.41 g, 69% yield).

[0409] 1 H NMR (400 MHz, CDCl3) δ = 9.99 (s, 1H), 7.92 (d, J = 7.7 Hz, 2H), 7.30 - 7.26 (m, 2H), 5.39 - 5.26 (m, 5H), 4.36 - 4.27 (m, 2H), 4.15 (ddd, J =1.1, 6.0, 12.0 Hz, 2H), 2.79 - 2.66 (m, 1H), 2.65 - 2.45 (m, 3H), 2.42 - 2.34(m, 1H), 2.30 (dt, J = 1.5, 7.5 Hz, 4H), 2.05 – 1.97 (m, 8H), 1.62 - 1.56 (m,4H), 1.40 - 1.20 (m, 40H), 1.14 (d, J = 4 Hz, 3H), 0.88 (t, J = 8Hz, 6H);

[0410] 13 C NMR (101 MHz, CDCl3) δ = 191.0, 173.4, 171.3, 170.2, 155.4, 134.2,131.4, 130.2, 129.8, 122.5, 69.5, 62.2, 40.7, 34.1, 32.1, 29.91, 29.85, 29.7,29.5, 29.31, 29.26, 29.2, 27.5, 27.4, 27.3, 25.0, 22.8, 19.9, 14.3.

[0411] Sodium borohydride (60.2 mg, 1.59 mmol) was added in portions to solution 2 (1.4 g, 1.56 mmol) in THF (14 mL) and MeOH (14 mL), and the reaction mixture was stirred at 0°C for 1 hour. The mixture was diluted with ethyl acetate (100 mL), water (100 mL), and brine (100 mL), dried over sodium sulfate, and concentrated. The substance was purified by normal phase chromatography (Biotage Isolera, 40 g Silia Sep filter cartridge, eluent 0-40% ethyl acetate / heptane, 16 CVs) to give 3 (1.14 g, 81% yield).

[0412] 1 H NMR (400 MHz, CDCl3) δ = 7.38 (d, J = 8.4 Hz, 2H), 7.07 (d, J = 8.4Hz, 2H), 5.42 - 5.23 (m, 5H), 4.69 (s, 2H), 4.31 (dt, J = 4.2, 11.9 Hz, 2H),4.14 (ddd, J = 2.2, 6.0, 12.0 Hz, 2H), 2.70 - 2.54 (m, 2H), 2.54 - 2.45 (m,2H), 2.41 - 2.27 (m, 5H), 2.08 - 1.94 (m, 8H), 1.70 - 1.53 (m, 4H), 1.28 (brd, J = 9.3 Hz, 40H), 1.16 - 1.11 (m, 3H), 0.88 (t, J = 6.8 Hz, 6H); No exchangeable protons were observed.

[0413] 13 C NMR (101 MHz, CDCl3) δ = 173.4, 171.4, 170.9, 150.1, 138.7, 130.2,129.9, 128.2, 121.8, 69.4, 64.9, 62.2, 40.7, 34.1, 32.0, 29.9, 29.8, 29.7,29.5, 29.31, 29.26, 29.2, 27.6, 27.4, 27.3, 25.0, 22.8, 19.9, 14.3.

[0414] A solution of 3 (1.14 g, 1.27 mmol), CBr4 (1.05 g, 3.17 mmol), and PPh3 (342 mg, 1.30 mmol) in dichloromethane (30 mL) was stirred at room temperature for 18 hours. The reaction mixture was concentrated, and the residue was purified by normal-phase chromatography (Biotage Isolera, 40 g Silia Sep filter cartridge) using 0–30% ethyl acetate / heptane as eluent for 12 CV cycles to give 4 (790 mg, 65% yield) as a colorless oil.

[0415] 1 H NMR (400 MHz, CDCl3) δ = 7.40 (d, J = 7.6 Hz, 2H), 7.06 (d, J = 7.6Hz, 2H), 5.39 - 5.27 (m, 5H), 4.48 (s, 2H), 4.37 - 4.27 (m, 2H), 4.14 (dd, J= 6.0, 12.0 Hz, 2H), 2.69 - 2.61 (m, 1H), 2.61 - 2.53 (m, 1H), 2.53 - 2.45(m, 2H), 2.41 - 2.27 (m, 5H), 2.14 - 1.88 (m, 8H), 1.60 (br t, J = 7.0 Hz,4H), 1.40 – 1.23 (m, 40H), 1.13 (d, J = 6.5 Hz, 3H), 0.95 – 0.81 (m, 6H);

[0416] 13 C NMR (101 MHz, CDCl3) δ = 173.4, 171.4, 170.6, 150.6, 135.5, 130.3,130.1, 129.8, 122.0, 69.4, 62.2, 40.7, 34.1, 32.7, 32.0, 29.9, 29.82, 29.78,29.6, 29.5, 29.4, 29.3, 29.23, 29.20, 27.6, 27.34, 27.29, 24.9, 22.8, 19.8,14.2.

[0417] TBAI (28.2 mg, 76.4 µmol) and tert-BuOK (25.1 mg, 224 µmol) was added to a solution of 2BL (60 mg, 149 µmol) in THF (3 mL) and stirred at room temperature for 10 minutes. 4 (123 mg, 134 µmol) was added and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate (30 mL) and washed with water (20 mL) and brine (30 mL). The organic layer was dried over MgSO4, filtered, and concentrated. The substance was purified by normal-phase chromatography (Biotage Isolera, 12 g SiliaSep filter cartridge) using 0-100% ethyl acetate / heptane as eluent to give 2BL-PHB-C5bMe-2-TG-oleate, compound I-20 (80 mg, 38% yield), as a yellow oil.

[0418] 1 H NMR (400 MHz, CDCl3) δ = 7.23 - 7.17 (m, 1H), 7.15 - 7.06 (m, 3H), 7.06 - 7.02 (m, 1H), 6.99 (d, J = 8.6 Hz, 2H), 6.36 (s, 1H), 5.40 - 5.24 (m,7H), 4.36 - 4.25 (m, 2H), 4.18 - 4.09 (m, 2H), 3.90 (br s, 1H), 3.52 - 3.38(m, 5H), 3.26 (br s, 1H), 3.07 (br d, J = 7.2 Hz, 1H), 2.92 (br d, J = 10.1Hz, 1H), 2.68 - 2.43 (m, 8H), 2.37 - 2.23 (m, 5H), 2.08 - 1.93 (m, 8H), 1.64- 1.53 (m, 4H), 1.37 - 1.21 (m, 43H), 1.17 (t, J = 7.1 Hz, 3H), 1.10 (d, J =6.5 Hz, 3H), 0.92 - 0.82 (m, 6H);

[0419] 13C NMR (101 MHz, CDCl3) δ = 173.4, 171.4, 170.7, 150.0, 135.1, 135.0,130.2, 129.9, 127.8, 126.0, 123.5, 121.9, 120.8, 113.4, 108.5, 69.4, 62.8,62.2, 56.1, 53.6, 47.8, 44.1, 42.2, 40.7, 40.4, 34.1, 32.0, 29.9, 29.8, 29.7,29.5, 29.30, 29.25, 29.2, 27.6, 27.4, 27.3, 25.0, 22.8, 19.8, 15.1, 14.3, 13.3.

[0420] Scheme 14: Synthesis of compound I-21 (2BL-CPHB-C5bMe-2-TG-oleate)

[0421]

[0422] Will N- Ethyl- N’ -(3-dimethylaminopropyl)carbodiimide hydrochloride (1.02 g, 5.34 mmol) and 4-dimethylaminopyridine (652 mg, 5.34 mmol) were added to a stirred solution of 4-hydroxybenzaldehyde (326 mg, 2.67 mmol) and Int-1 (2 g, 2.67 mmol) in dichloromethane (40 mL). The mixture was stirred at room temperature for 18 hours, concentrated under reduced pressure, and purified by normal-phase chromatography (Biotage Isolera, 80 g SiliaSep filter cartridge) using 0–20% ethyl acetate / heptane as eluent to give 1 (1.82 g, 77% yield).

[0423] 1 H NMR (400 MHz, CDCl3): δ[ppm] = 10.00 (s, 1H), 7.92 (d, J = 8.6 Hz, 2H), 7.28 (d, J = 8.6 Hz, 2H), 5.41 - 5.25 (m, 5H), 4.33 (ddd, J = 1.5, 4.1, 11.9Hz, 2H), 4.15 (dd, J = 6.0, 11.9 Hz, 2H), 2.75 - 2.68 (m, 1H), 2.65 - 2.45 (m,3H), 2.43 - 2.35 (m, 1H), 2.31 (t,J = 7.5 Hz, 4H), 2.04 - 1.96 (m, 8H), 1.63 -1.57 (m, 4H), 1.28 (br d, J = 9.3 Hz, 40H), 1.15 (d, J = 6.4 Hz, 3H), 0.88 (t, J =6.8 Hz, 6H).

[0424] 13 C NMR (100 MHz, CDCl3): δ [ppm] =191.0, 173.4, 171.3, 170.2, 155.4,134.2, 131.4, 130.2, 129.9, 122.5, 69.5, 62.2, 40.67, 40.65, 34.2, 32.1,29.91, 29.85, 29.7, 29.5, 29.32, 29.26, 29.24, 27.5, 27.4, 27.3, 25.0, 22.8,19.9, 14.3.

[0425] Sodium borohydride (85 mg, 2.25 mmol) was added in portions to 1 (1.82 g, 2.13 mmol) in a stirred solution of THF (20 mL) and methanol (20 mL) and stirred at room temperature for 3 hours. The mixture was diluted with water (80 mL) and extracted into ethyl acetate (2 × 70 mL). The combined organic extracts were washed with brine (80 mL), dried over sodium sulfate, filtered, and concentrated. The substance was purified by normal phase chromatography (Biotage Isolera, 80 g SiliaSep filter cartridge) using an eluent of 0–100% ethyl acetate / heptane to give 2 (790 mg, 41% yield) as a colorless oil.

[0426] 1 H NMR (400 MHz, CDCl3): δ[ppm] =7.38 (d, J = 8.6 Hz, 2H), 7.08 - 7.05 (m,2H), 5.39 - 5.24 (m, 5H), 4.69 (d, J = 5.9 Hz, 2H), 4.31 (td, J2.05 - 1.96 (m, 8H), 1.66 - 1.56 (m, 4H), 1.36 - 1.21 (m, 40H), 1.14 (d, J = 6.5 Hz, 3H), 0.88 (t, J = 6.7 Hz, 6H); No exchangeable hydroxyl protons were observed.

[0427] At 0°C, nitrobenzene chloroformate (47.1 mg, 234 µmol) was added to a stirred solution of 2 (213 mg, 234 µmol) in DCM (4 mL), followed by the addition of N,N A solution of diisopropylethylamine (167 µL, 959 µmol) was prepared. The reaction mixture was warmed to room temperature and stirred for 18 hours. The mixture was diluted with water (10 mL) and DCM (6 mL). The organic layer was extracted, concentrated, and purified by normal-phase chromatography (Biotage Isolera, 12 g SiliaSep filter cartridge) using 0–20% ethyl acetate / heptane to give 3 (123 mg, 32% yield) as a colorless oil.

[0428] 1 H NMR (400MHz, CDCl3): δ[ppm] =8.28 (d, J = 9.3 Hz, 2H), 7.47 (d, J = 8.6Hz, 2H), 7.43 - 7.35 (m, 2H), 7.16 - 7.04 (m, 2H), 5.40 - 5.25 (m, 5H), 5.14(s, 2H), 4.37 - 4.27 (m, 2H), 4.14 (br dd, J = 6.0, 11.9 Hz, 2H), 2.73 - 2.63(m, 1H), 2.63 - 2.54 (m, 1H), 2.54 - 2.44 (m, 2H), 2.42 - 2.34 (m, 1H), 2.34- 2.28 (m, 4H), 2.06 - 1.94 (m, 8H), 1.60 (br t, J= 6.6 Hz, 4H), 1.36 - 1.20(m, 40H), 1.13 (dd, J = 4.3, 6.5 Hz, 3H), 0.87 (t, J = 6.5 Hz, 6H).

[0429] Sodium hydride (5.79 mg, 145 µmol, 60% dispersion in mineral oil) was added to a solution of 2BL (48.5 mg, 121 µmol) in THF (3 mL) and stirred for 30 minutes. A solution of 3 dissolved in THF (1 mL) was added to the mixture. The mixture was warmed to room temperature and stirred for 18 hours. The mixture was diluted with ethyl acetate (15 mL) and washed with water (2 × 15 mL) and brine (2 × 15 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to a yellow oil. The substance was purified by normal phase chromatography (Biotage Isolera, 12 g SiliaSep filter cartridge) using 0–100% ethyl acetate + 0.5% triethylamine / heptane eluent to give a deep yellow gel-like 2BL-CPHB-C5bMe-2-TG-oleate, compound I-21 (60 mg, 32% yield).

[0430] Example 3: Metabolic Identification (MetID) Determination in Human Hepatocytes, Rat Hepatocytes, and Human Liver Microsomes

[0431] To better understand the clearance mechanism and biotransformation capacity, resolve the presence of potential active or reactive metabolites, and observe disproportionate or unique metabolites of bromo-lysergic acid diethylamide (2-bromo-LSD), a MetID assay was performed. Samples used for MetID included human hepatocytes, rat hepatocytes, and human liver microsomal S9 fractions. Hepatocytes were prepared by thawing cryopreserved cells in inVitroGRO KHB buffer (IVT) to a concentration of 1,000,000 cells / mL. After pre-incubation at 37°C for 5 min, the hepatocytes were also incubated with bromo-lysergic acid diethylamide (2-bromo-LSD) (final concentration 10 μM, containing <0.2% organic matter) at 37°C for 3 h. Next, 100 μL of the incubation mixture was removed after incubation and quenched with 100 μL of acetonitrile, then vortexed and centrifuged at 13,000 rpm for a total of 5 min. After centrifugation, 175 μL of the supernatant was added to a 96-well plate and dried with nitrogen to a 100 μL sample for use in liquid chromatography-high resolution mass spectrometry.

[0432] Similarly, liver microsomal incubation was performed. 1 mg / mL cryopreserved liver microsomal S9 fractions were incubated in phosphate-buffered saline (pH 7.4, 100 mM, containing 3 mM MgCl2) at 37°C.o Incubate at C. After 5 minutes, add 1 mM NADPH to initiate the reaction and incubate the microsomes with bromo-2-bromo-LSD for one hour. The final concentration of bromo-2-bromo-LSD was 10 μM, containing <0.2% organic matter. Next, after incubation, remove 100 μL of the incubation mixture and quench with 100 μL of acetonitrile, then vortex and centrifuge at 10,000 rpm for a total of 5 minutes. After centrifugation, collect 170 μL of the supernatant and dry it to 100 μL of sample for ultra-high performance liquid chromatography-high resolution mass spectrometry.

[0433] In both types of incubation, 5.0 μM verapamil was used as a positive control, while the negative control consisted only of incubation buffer. LC / MS data were processed manually using the programs Xcalibur, Freestyle, and Compound Discoverer. Results are summarized in Table 2 below.

[0434] MetID results determined that lysergic acid diethylamide (LSD) was not a major metabolite. Figure 1 (Table 2). MetID was also performed because the main metabolite of bromolybdate diethylamine (2-bromo-LSD) is likely 5-HT. 2B Strong receptor agonists, which may lead to fibrotic side effects with chronic administration. Due to the stability of the bromine atom, none of the major metabolites identified in the MetID results appear likely to activate 5-HT. 2B The few debrominated metabolites are rapidly oxidized, saturating the ring structure. Therefore, these metabolites are also unlikely to activate 5-HT. 2B Receptors.

[0435] Table 2: Metabolites of bromolybdate diethylamine identified in human liver microsomes (Human_LM), rat hepatocytes (Rat), and human hepatocytes (Human).

[0436]

[0437] Example 4: In vitro hydrolysis of lipoprotein prodrugs of 2-bromo-LSD

[0438] The selectivity of prodrug hydrolysis for recombinant human esterases rhCES1, rhCES2, and rhBChE was evaluated. For each recombinant human esterase, 50 µM of prodrug was incubated in 10 mg / mL bovine serum albumin (BSA), 300 units / mL lipoprotein lipase, and 0.5 µg / mL recombinant human esterase. The reaction was initiated by adding the prodrug. After incubation at 37°C for 30 min, the reaction was quenched by adding cold acetonitrile (3:1 acetonitrile:reaction volume) containing an internal standard. The samples were vortexed and centrifuged, and the supernatant was transferred and diluted in 0.1% formic acid (1:3 supernatant to 0.1% formic acid). The samples were analyzed by LC-MS, and the percentage of 2-bromo-LSD was calculated using a standard curve of 2-bromo-LSD in BSA (10 mg / mL in PBS).

[0439] To determine the rate of 2-bromo-LSD release catalyzed by each esterase, the reactants were prepared as described above. The mixture was incubated at 37°C, and 50 µL aliquots were removed from the incubation at 5, 10, 15, 20, and 30 minutes. For each aliquot, the reactants were quenched by adding 150 µL of cold acetonitrile containing d3-LSD (40 ng / mL). The samples were vortexed and centrifuged, and the supernatant was transferred and diluted in 0.1% formic acid (1:3 supernatant to 0.1% formic acid). The samples were analyzed by LC-MS, and the percentage of 2-bromo-LSD was calculated using a standard curve of 2-bromo-LSD in BSA (10 mg / mL in PBS).

[0440] Example 5: In vitro stability of lipoprotein prodrug of 2-bromo-LSD

[0441] The stability of the lipoprotein prodrug of 2-bromo-LSD was assessed in the presence and absence of recombinant human monoacylglycerol lipase (rhMAGL). 50 µM of the prodrug was incubated in 10 mg / mL BSA supplemented with lipoprotein lipase (300 units / mL) and rhMAGL (0 µg / mL–0.5 µg / mL). The MAGL concentration could be adjusted based on observed activity. The reaction was initiated by adding the prodrug and incubated at 37°C for 180 min. Aliquots were removed at 30, 60, 120, and 180 min, and the reaction mixture was quenched by adding cold acetonitrile (1:3 sample to acetonitrile) containing an internal standard. The samples were vortexed and centrifuged, and the supernatant was transferred and diluted in 0.1% formic acid water (1:3 supernatant to 0.1% formic acid water). The samples were analyzed by LC-MS. The monoacylglycerol stability was calculated using the following equation:

[0442]

[0443]

[0444]

[0445] The monoglyceride stability of the lipid prodrug is shown in Table 3. Stability is calculated on a scale of -10 to 10, where 10 represents 100% remaining monoglycerides ( Right now (highest stability), -10 indicates 100% monoglyceride loss ( Right now (Minimum stability). Monoglyceride stability is represented by the following categories, where A is a stability score of 5 to 10, B is a stability score of 0 to 5, C is a stability score of -5 to 0, and D is a stability score of -5 to -10.

[0446] Table 3: Stability of monoacylglycerol esters of lipid prodrugs

[0447]

[0448] Example 6: In vitro release of 2-bromo-LSD from a lipid prodrug in plasma

[0449] use in vitro The assay was performed to determine the release of 2-bromo-LSD in plasma supplemented with lipoprotein lipase. Lipoprotein lipase (LPL) is a key enzyme in the hydrolysis of lipoprotein-associated triglycerides in systemic circulation and is expected to participate in the breakdown of 2-bromo-LSD from the triglyceride backbone in plasma. Right now Releases free 2-bromo-LSD.

[0450] 40 µM of prodrug was incubated with human plasma containing 200 units / mL–12,000 units / mL LPL (LPL concentration determined experimentally based on the units required to produce the maximum monoglyceride concentration after initiation). The sample was incubated at 37°C for 0–180 minutes. The reactants were quenched with MeCN containing an internal standard, vortexed, and centrifuged at 4000 x g for 10 minutes to precipitate the protein. The supernatant was analyzed by LC-MS / MS, and potential hydrolysis products, namely monoglyceride form, acid form, and free 2-bromo-LSD, were detected. The concentration of free 2-bromo-LSD was calculated using a standard curve of 2-bromo-LSD. The percentage of free 2-bromo-LSD in the tested prodrugs is shown in Table 4, where +++ indicates that the percentage of 2-bromo-LSD is greater than 25%, indicating that the level of 2-bromo-LSD released from the prodrug is sufficient, ++ indicates that the percentage of 2-bromo-LSD released is between 5% and 25%, and + indicates that the percentage of 2-bromo-LSD is less than 5%.

[0451] Table 4: Percentage of 2-bromo-LSD released from human plasma

[0452]

[0453] Example 7: In vitro release of 2-bromo-LSD from whole blood

[0454] The release of free 2-bromo-LSD from the prodrug backbone was evaluated in whole blood supplemented with lipoprotein lipase. Lipoprotein lipase (LPL) is a key enzyme in the hydrolysis of lipoprotein-associated triglycerides in systemic circulation and is expected to be involved in the lipid breakdown of 2-bromo-LSD from the triglyceride backbone in plasma. Right now Releases free 2-bromo-LSD.

[0455] A prodrug stock solution was prepared at a concentration of 10 mM in a 7:3 MeCN:IPA mixture. To initiate hydrolysis, whole blood (stored at 4°C) of human or NHP doped with LPL (6000 IU / mL) and ketoprofen (400 ng / mL) was added to human or NHP plasma containing 80 µM of the prodrug. The sample was incubated at room temperature and mixed by inversion. Aliquots were removed from each reaction at 5, 30, 60, and 120 minutes and centrifuged at 2000 x g for 10 minutes to collect plasma. 30 µL of plasma was added to 300 µL of an organic solvent containing an internal standard (IPA:MeOH:MeCN:FA in a 1:1:1:0.1 ratio). The mixture was vortexed and centrifuged at 4000 x g for 10 minutes to precipitate proteins. The potential hydrolysis products of the supernatant were then analyzed by HPLC-MS / MS, including the monoglyceride form of the prodrug, the acid form of the prodrug, and free 2-bromo-LSD.

[0456] Example 8: Pharmacokinetics of 2-bromo-LSD prodrug in rats

[0457] Systemic plasma exposure to 2-bromo-LSD induced by oral administration of a lipid prodrug was determined in rats. (See WO 2016 / 023082, WO 2017 / 041139, and Trevaskis NL). people, The lipid-based formulations for preparing lipid prodrugs or control compounds described in Pharmaceutical Research (2005) 22(11):1863-1870 are incorporated herein by reference.

[0458] Mix 1 mg or 2 mg of 2-bromo-LSD or its prodrug, 40 mg of oleic acid, and 25 mg of polysorbate 80 in a glass vial and incubate at 37°C for at least one hour and up to 18 hours to reach equilibration. Add 2 mL of the aqueous phase of phosphate-buffered saline (PBS, pH 7.4) to the vial. Emulsify the formulation by sonication at room temperature for 2 minutes using a probe-tip sonicator with 10-second pulses (10 seconds on, 10 seconds off). Maintain the vial temperature below 40°C. Verify the formulation concentration by HPLC-MS or HPLC-UV.

[0459] As a control, 2-bromo-LSD was dissolved in DMSO and diluted 10-fold in a 20% aqueous solution of hydroxypropyl-β-cyclodextrin (HPCD) for intravenous (IV) or oral (PO) administration.

[0460] Male Sprague-Dawley rats (220 g–320 g) were fasted overnight but allowed free access to water. The following morning, rats were administered either the prodrug formulation or a 2-bromo-LSD control formulation orally. As an additional control, 2-bromo-LSD was administered via IV bolus. The prodrug was administered at 4 mg / kg and normalized to a 1 mg / kg 2-bromo-LSD equivalent. Following oral administration, blood samples (0.23 mL) were collected via tail vein or other suitable venous puncture approximately -5 minutes to 24 hours post-administration. Blood samples were collected in KEDTA tubes pre-filled with sufficient inhibitor solutions such that the final concentration of each inhibitor after sample collection was 40 µg / mL of orlistat and 100 µM of JZL-184 and GR148672X inhibitors. Plasma was separated by centrifugation and stored at -80°C before analysis by HPLC-MS / MS. Table 5 provides plasma exposure to 2-bromo-LSD in rats.

[0461] Table 5: Plasma exposure of 2-bromo-LSD in rats

[0462]

[0463] Example 9: Pharmacokinetics of 2-bromo-LSD prodrug in dogs

[0464] Systemic plasma exposure to 2-bromo-LSD induced by oral administration of a lipid prodrug was determined in dogs. Male beagles were fasted for 12 hours to 30 minutes prior to drug administration. For the feeding status study, dogs received approximately 20 g of high-fat dog food (containing approximately 34% fat) by hand administration, followed by 10 mL of water and 100 g of standard canned dog food (containing approximately 2.5% fat) 30 minutes prior to drug administration. Throughout the study, systemic plasma exposure to 2-bromo-LSD induced by oral administration of a lipid prodrug was determined in dogs. randomDrinking water. The prodrug can be prepared in a suitable SEDDS formulation, such as 25% w / w sesame oil, 27% w / w glyceryl monooleate, and 48% polyethoxylated castor oil. The formulation can be filled into gelatin capsules or dispersed in water for oral tube feeding. As a control, 2-bromo-LSD was dissolved in DMSO and diluted 10-fold in a 20% aqueous solution of hydroxypropyl-β-cyclodextrin (HPCD).

[0465] The compound can be administered orally by placing the capsule behind the throat, closing the mouth, and stimulating swallowing. 50 mL of water will be administered orally via syringe. Following oral administration, blood samples (approximately 1.5 mL each) will be obtained via venipuncture of the cephalic vein 5 minutes before administration and up to 120 hours after administration. Plasma will be separated by centrifugation, and aliquots of each plasma sample will be stored at -80°C. As a control, 2-bromo-LSD can be administered intravenously by infusion (over 5 minutes) or rapid concentration.

[0466] Example 10: Pharmacokinetics of 2-bromo-LSD prodrug in non-human primates

[0467] Systemic plasma exposure to 2-bromo-LSD induced by oral administration of a lipid prodrug was determined in nonhuman primates. The lipid prodrug or 2-bromo-LSD was formulated in a SEDDS formulation of 25% w / w sesame oil, 27% w / w glyceryl monooleate, and 48% polyethoxylated castor oil. The formulation could be filled into gelatin capsules or dispersed in water for oral tube feeding. As a control, 2-bromo-LSD was dissolved in DMSO and diluted 10-fold in a 20% aqueous solution of hydroxypropyl-β-cyclodextrin (HPCD).

[0468] The cynomolgus monkeys were fasted for 12 hours prior to administration. In the feeding state study, NHPs received 30 mL of Ensure shake (available from Abbott) approximately 30 minutes before administration, followed by 10 mL of water. Water intake was permitted at will throughout the study. The formulation dispersed in water was administered orally to the NHPs via oral tube feeding and flushed with 50 mL of water after delivery. For the prodrug, the NHPs were administered 1 mg / kg equivalent of 2-bromo-LSD. As a comparative control, 2-bromo-LSD was administered via oral tube feeding. To calculate bioavailability, 2-bromo-LSD was administered via IV bolus.

[0469] Following oral administration, blood samples (0.6 mL) were obtained via venipuncture at the cephalic vein or other suitable sampling site approximately 5 minutes to 120 hours after administration. Blood samples were collected in K2EDTA tubes pre-filled with sufficient inhibitor solution to achieve a final concentration of 40 µg / mL of orlistat and 100 µM of JZL-184, GR148672X, and rivastigmine inhibitors at the time of sample collection. Plasma was separated by centrifugation and stored at -80°C prior to analysis. Oral bioavailability was calculated by administering 2-bromo-LSD via oral tube feeding or IV infusion as a comparative control.

[0470] Plasma samples were analyzed by LC-MS / MS to measure free and combined 2-bromo-LSD. To determine free 2-bromo-LSD, samples were extracted using protein precipitation and quantified against a standard curve for 2-bromo-LSD. As used herein, the term "combined 2-bromo-LSD" refers to 2-bromo-LSD detected as free 2-bromo-LSD, an unhydrolyzed prodrug of 2-bromo-LSD, and a hydrolyzed intermediate of the 2-bromo-LSD prodrug. For the analysis of combined 2-bromo-LSD, samples were diluted in IPA and incubated at room temperature for 2 hours after the addition of KOH (20 mg / mL). This incubation step hydrolyzed the prodrug or prodrug intermediate to 2-bromo-LSD. After the reaction was neutralized, the 2-bromo-LSD level was quantified against a standard curve for the corresponding prodrug. Table 6 provides plasma exposure of 2-bromo-LSD in male NHPs, and Table 7 provides plasma exposure of 2-bromo-LSD in female NHPs. Table 8 provides the combined levels of 2-bromo-LSD in male NHPs, and Table 9 provides the combined levels of 2-bromo-LSD in female NHPs.

[0471] Figures 2A to 2D It has been proven that in aqueous formulations ( Figure 2A SEDDS lipid formulations ( Figure 2B (in or as a prodrug of this disclosure) Figure 2C and Figure 2D Systemic exposure to 2-bromo-LSD administered.

[0472] Label 6: Plasma exposure of 2-bromo-LSD in male NHPs

[0473]

[0474] Table 7: Plasma exposure of 2-bromo-LSD in female NHPs

[0475]

[0476] Table 8: Plasma exposure of combined 2-bromo-LSD in male NHPs

[0477]

[0478] Table 9: Plasma exposure of combined 2-bromo-LSD in female NHPs

[0479]

Claims

1. A compound of formula I: Equation (I) Or its pharmaceutically acceptable salt, wherein: R 1 and R 2 are each independently hydrogen or -C(O)R 3 ; Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 2-37 hydrocarbon chain; X is -O-; R 4 and R 5 Each of the C atoms is independently hydrogen or optionally substituted with one, two, three, four, five, or six deuterium or halogen atoms. 1-4 aliphatic group, or R 4 and R 5 Together with the carbons they are attached to, they form C3-C8 cycloalkyl groups; M is absent or is a self-exfoliating group; n is 0-18; and m and q are each independently 0-6.

2. The compound of claim 1, wherein R 1 and R 2 Each is independently -C(O)R 3 .

3. The compound of claim 1 or 2, wherein each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

4. The compound according to any one of claims 1 to 3, wherein each R 3 It is a C7 hydrocarbon chain.

5. The compound according to any one of claims 1 to 4, wherein m and q are each independently 0 or 1.

6. The compound according to any one of claims 1 to 5, wherein n is 0-6.

7. The compound according to any one of claims 1 to 6, wherein R 4 and R 5 Each is either hydrogen or methyl.

8. The compound according to any one of claims 1 to 7, wherein M is selected from... , , , , , , , , , , , , , and .

9. The compound according to any one of claims 1 to 8, wherein M is , , , , , , or .

10. The compound according to any one of claims 1 to 9, wherein the compound of formula I is selected from Table 1.

11. A compound of formula II: (Formula II) Where R 1 and R 2 Each is independently -C(O)R 3 ;and Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

12. The compound of claim 11, wherein each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain.

13. The compound of claim 11 or 12, wherein each R 3 It is a saturated C7 hydrocarbon chain.

14. The compound according to any one of claims 11 to 13, wherein the compound is （I-19）。 15. A compound of formula III: (Formula III) Where R 1 and R 2 Each is independently -C(O)R 3 ;and Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

16. The compound of claim 15, wherein each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain.

17. The compound of claim 15 or 16, wherein each R 3 It is a saturated C7 hydrocarbon chain.

18. The compound of any one of claims 15 to 18, wherein the compound is: （I-16）。 19. A compound of formula IV: (Form IV) Where R 1 and R 2 Each is independently -C(O)R 3 ;and Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

20. The compound of claim 19, wherein each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain.

21. The compound of claim 19 or 20, wherein each R 3 It is a saturated C7 hydrocarbon chain.

22. The compound according to any one of claims 19 to 21, wherein the compound is: （I-9）。 23. A compound of formula V: (Form V) Where R 1 and R 2 Each is independently -C(O)R 3 ;and Each R 3 C can be saturated or unsaturated, straight or branched, or optionally substituted. 3-17 Hydrocarbon chain.

24. The compound of claim 23, wherein each R 3 Independently, it is a straight-chain C that is either saturated or unsaturated. 3-17 Hydrocarbon chain.

25. The compound of claim 23 or 24, wherein each R 3 It is a saturated C7 hydrocarbon chain.

26. The compound according to any one of claims 23 to 25, wherein the compound is: （I-18）。 27. A pharmaceutically acceptable composition comprising the compound as described in any one of claims 1 to 26 and a pharmaceutically acceptable carrier, adjuvant, or mediator.

28. The pharmaceutically acceptable composition of claim 27, administered orally to a subject in need.

29. A method of treating cluster headaches in a patient, comprising orally administering to a patient in need a compound as described in any one of claims 1 to 26 or a pharmaceutically acceptable composition as described in claim 27.

30. A method of treating a patient's migraine, comprising orally administering to a patient in need a compound as described in any one of claims 1 to 26 or a pharmaceutically acceptable composition as described in claim 27.

31. A method of treating a patient’s mood disorder, comprising orally administering to a patient in need a compound as described in any one of claims 1 to 26 or a pharmaceutically acceptable composition as described in claim 27.

32. The method of claim 31, wherein the mood disorder is depression, bipolar disorder, premenstrual syndrome irritability disorder (PMDD), intermittent outburst disorder (IED), post-traumatic stress disorder (PTSD), or substance-induced mood disorder.

33. A method of treating a patient with an anxiety disorder, comprising orally administering to a patient in need a compound as described in any one of claims 1 to 26 or a pharmaceutically acceptable composition as described in claim 27.

34. The method of any one of claims 29 to 33, further comprising the step of administering an additional therapeutic agent.

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